An estrogen receptor protein targeted degradation chimeric derivative based on the structure of estratriene and a preparation method and application thereof

Abstract

The application discloses an estrogen receptor protein targeted degradation chimera derivative based on an oestranetriene structure and a preparation method and application thereof, and belongs to the technical field of drug synthesis and application. The estrogen receptor protein targeted degradation chimera derivative based on the oestranetriene structure not only has a good inhibitory effect on estrogen-induced signal transduction, but also has a good degradation effect on ERa. The estrogen receptor protein targeted degradation chimera derivative has good pharmacokinetics, bioavailability and in-vivo drug efficacy, and can be developed into a new estrogen receptor degrading agent drug.

Description

Technical Field

[0001] This application relates to a chimeric derivative of estrogen receptor protein targeted degradation based on an estradiol structure, its preparation method and application, belonging to the field of drug synthesis and application technology. Background Technology

[0002] Breast cancer is the most common malignant disease among women worldwide, and about 60% of breast cancer tumors are estrogen receptor alpha (ER) positive at diagnosis. Estrogen receptors play a crucial role in the progression of the disease; for example, ER protein levels are elevated in the pre-malignant stage and during malignant lesions of the breast compared to normal tissue. + Tumor growth is significantly correlated with anti-estrogenic therapy; anti-estrogenic therapy blocks estrogen receptor binding sites, thereby inhibiting cancer cell growth. According to estimates from the National Cancer Institute (NCI), approximately 250,000 women were diagnosed with breast cancer in 2017, and more than 40,000 of them died from it. + / HER2 - Breast cancer accounts for approximately 70% of all breast cancer cases. These patients can initially be treated with endocrine blockade therapy, but as the disease progresses, the tumor develops resistance to this therapy and accumulates numerous mutations, making treatment more difficult.

[0003] In recent years, the incidence of breast cancer has been on the rise, and it now ranks first among malignant tumors in women. Approximately 70% of breast cancer patients present with estrogen receptor (ER)-positive breast cancer. Therefore, ER is an important target in breast cancer drug research. To date, the primary intervention for treating ER-positive breast cancer remains endocrine therapy, which suppresses estrogen levels associated with breast cancer. This mainly includes selective estrogen receptor modulators (such as tamoxifen and raloxifene), aromatase inhibitors (such as letrozole and exemestane), and selective ER downregulators. Although endocrine therapy has achieved great success in treating breast cancer patients, it has drawbacks such as toxic side effects, long treatment duration, and drug resistance.

[0004] PROTAC (Protein Hydrolysis Targeted Chip) technology, proposed by Professor Craig M. Crews' team, is a cutting-edge targeted therapy strategy. PROTACs are hybrid bifunctional molecules with a ligand at one end that binds to an E3 ligase, and a ligand at the other end that binds to a target protein (POI), with a linker in the middle. The mechanism of action involves PROTACs recruiting the target protein to the vicinity of the E3 ligase, leading to ubiquitination and subsequent degradation of the target protein via the ubiquitin-proteasome pathway, ultimately achieving therapeutic effects. By degrading ER (erucic acid) in breast cancer cells, PROTACs can effectively overcome drug resistance and represent a promising anti-breast cancer drug.

[0005] Currently, there are relatively few types of estrogen receptor protein-targeting degradation chimeric derivatives. Developing a variety of novel estrogen receptor protein-targeting degradation chimerics to meet application needs has always been a research focus, especially on preparing estrogen receptor protein-targeting degradation chimerics with higher activity and stronger specificity. Furthermore, the currently marketed drug fulvestrant can only degrade 50% of the estrogen receptor, exhibiting a low degradation upper limit, which restricts the applications of this type of compound. Summary of the Invention

[0006] To address the aforementioned issues, a chimeric derivative of estrogen receptor protein targeting degradation based on an estradiol structure is provided, having a general formula (I) structure. This compound can solve the problems existing in the prior art.

[0007] According to the first aspect of this application, an estrogen receptor protein targeted degradation chimeric derivative based on an estradiol structure is provided, having a structure of general formula (I): ABC formula (I);

[0008] Where A is an E3 ligase ligand, selected from one of general formulas (II-1), (II-2), or (II-3):

[0009] Equation (Ⅱ-1);

[0010] Equation (Ⅱ-2);

[0011] Equation (Ⅱ-3);

[0012] * indicates a connection site, n=0 or 1, m=0 or 1, in formula (II-1), general formula (II-2) and general formula (II-3) R1, R2 and R3 are each independently one of: H, OH, F, Cl, Br, NH2, CH3 or OCH3, and R4 is hydrogen or methyl.

[0013] B is at least one of rigid linker, flexible linker, and bridging group;

[0014] C has the structure of general formula (Ⅲ):

[0015] Formula (III).

[0016] The estrogen receptor protein-targeting chimeric derivative of general formula (I) based on the estradiol structure serves as a bifunctional degrader. It retains the halogenated long chain portion of fulvestrant that exerts its degradation mechanism, and its degradation mechanism after binding to the E3 ligase ligand is that of a bifunctional stimulatory receptor degrader, acting as a PROTAC and estradiol analogue.

[0017] Compared to fulvestrant, the estrogen receptor protein-targeting chimeric derivative of general formula (I) exhibits higher estrogen receptor degradation efficacy in MCF-7 cells, potentially overcoming the limitation of fulvestrant in degrading only 50% of estrogen receptors. This could lead to more effective estrogen receptor degradation, achieving a higher degradation limit and better efficacy.

[0018] Optionally, the estrogen receptor protein targeted degradation chimeric derivative based on the estradiol structure includes its cis-trans isomer and racemic isomer.

[0019] Preferably, the E3 ligase ligand is selected from lenalidomide, thalidomide, 4-hydroxy-thalidomide, 5-hydroxy-thalidomide, pomalidomide, 3-(1-oxoisoindolin-2-yl)piperidin-2,6-dione, 4-amino-4-oxo-2-(1-oxoisoindolin-2-yl)butyric acid, or 2-(1-methyl-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione, or derivatives thereof and their isomers.

[0020] Preferably, the E3 ligase ligand in formula (I) is lenalidomide (IIa):

[0021] (IIa).

[0022] The linker-linking site (*) of lenalidomide (IIa) is one of the following:

[0023] (IIa-1), (IIa-2),

[0024] (IIa-3), or (IIa-4)

[0025] (Where * indicates a connection site. The same applies below).

[0026] The *NH- group in formula (IIa-1) may also be part of the Linker group or a terminal group (-NH-), as hereinafter referred to below. That is, as shown in the following formula:

[0027] (IIa-1), where -L'-NH- is either -Linker- or -L- (as a divalent linker). The same applies below.

[0028] Alternatively, the E3 ligase ligand in formula (I) is thalidomide (IIb):

[0029] (IIb).

[0030] The linker site (*) for thalidomide (IIb) is one of the following:

[0031] (IIb-1), or (IIb-2).

[0032] Alternatively, the E3 ligase ligand in formula (I) is 4-hydroxy-thalidomide (IIc-1) or 5-hydroxy-thalidomide (IIc-2):

[0033] (IIc-1), or (IIc-2).

[0034] The site at which the 4-hydroxy-thalidomide (IIc-1) is linked to the linker is one of the following:

[0035] (IIc-1a), (IIc-1b),

[0036] (IIc-1c), or (IIc-1d).

[0037] The linker site (*) for the 5-hydroxy-thalidomide (IIc-2) is one of the following:

[0038] (IIc-2a), (IIc-2b),

[0039] (IIc-2c), or (IIc-2d).

[0040] The *O- group can also be part of the linker group or a terminal group (-O-), as hereinafter referred to below. That is, as shown in the following formula:

[0041] Where -L'-O- is either -Linker- or -L- (as a divalent linker). The same applies below.

[0042] Alternatively, the E3 ligase ligand in formula (I) is pomalidomide (IId):

[0043] (IId-1), or (IId-2).

[0044] The linker site for pomalidomide (IId-1) is one of the following:

[0045] (IId-1a), (IId-1b),

[0046] (IId-1c), or (IId-1d).

[0047] The *NH- group may also be part of the linker group or a terminal group (-NH-), as hereinafter referred to below. That is, as shown in the following formula:

[0048] (IId-1), where -L'-NH- is either -Linker- or -L- (as a divalent linker). The same applies below.

[0049] The linker site for pomalidomide (IId-2) is one of the following:

[0050] (IId-2a), (IId-2b),

[0051] (IId-2c), or (IId-2d).

[0052] The *NH- group may also be part of the linker group or a terminal group (-NH-), as hereinafter referred to below. That is, as shown in the following formula:

[0053] (IId-2), where -L'-NH- is either -Linker- or -L- (as a divalent linker). The same applies below.

[0054] Alternatively, the E3 ligase ligand in formula (I) is 3-(1-oxoisoindololin-2-yl)piperidine-2,6-dione (IIe):

[0055] (IIe).

[0056] The site (*) or manner by which the 3-(1-oxoisoindolin-2-yl)piperidine-2,6-dione (IIe) is linked to the linker is, or, the E3 ligase ligand in formula (I) is, for example, one of the following:

[0057] (IIe-1), (IIe-2),

[0058] (IIe-3), or (IIe-4).

[0059] Alternatively, the E3 ligase ligand in formula (I) is 4-amino-4-oxo-2-(1-oxoisoindol-2-yl)butyric acid (IIf):

[0060] (IIf).

[0061] The linker-linking site (*) of the 4-amino-4-oxo-2-(1-oxoisoindol-2-yl)butyric acid (IIf) is one of the following:

[0062] (IIf-1), (IIf-2),

[0063] (IIf-3), or (IIf-4).

[0064] Alternatively, the E3 ligase ligand in formula (I) is 2-(1-methyl-2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (IIg):

[0065] (IIg).

[0066] The linker-linking site (*) of the 2-(1-methyl-2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (IIg) is one of the following:

[0067] (IIg-1), or (IIg-2).

[0068] It is at least one of a rigid linker, a flexible linker, and a bridging group; when it is a rigid linker, it is selected from one of the following formulas:

[0069] This is the connection site.

[0070] When it is a flexible connecting base, it is selected from one of the following formulas:

[0071]

[0072] When it is a combination of flexible and rigid connecting groups, it is selected from one of the following formulas:

[0073]

[0074]

[0075]

[0076]

[0077] In this application, n=0-5 means n=0, 1, 2, 3, 4 or 5. Similarly, n=2-10 means n=2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0078] Optionally, the estrogen receptor protein-targeting degradation chimeric derivative based on the estradiol structure has one of the following structural formulas:

[0079] ; ; ;

[0080] ;

[0081] ;

[0082] ;

[0083] ; ; ; ;

[0084] .

[0085] According to a second aspect of this application, a method for preparing the estrogen receptor protein-targeted degradation chimeric derivative based on the estradiol structure described in any one of the above claims is provided, comprising the following steps:

[0086] (1) Place fulvestrant in a solvent, add an acid-binding agent and lower the temperature to 1-5℃, then add a monomer containing halogen atoms and acyl halide groups to react and obtain compound 2;

[0087] (2) Compound 2 is reacted sequentially with a rigid linker and an E3 ligase ligand A with a flexible linker to obtain an estrogen receptor protein-targeting degradation chimeric derivative of general formula (I) based on an estradiol structure. The E3 ligase ligand A with a flexible linker has a carboxyl group or a ketone carbonyl group at its end; or

[0088] Compound 2 was reacted with E3 ligase ligand B with a flexible linker to obtain an estrogen receptor protein targeted degradation chimeric derivative of general formula (I) based on an estradiol structure, wherein the E3 ligase ligand B with the flexible linker has an amino group at its end.

[0089] In step (1) above, the hydroxyl group of fulvestrant and the acyl halide group of the monomer containing halogen atoms and acyl halide groups react under the action of an acid-binding agent, thereby causing compound 2 to carry halogen atoms at its end.

[0090] In the two reactions of step (2), the first reaction is a condensation reaction between the halogen atom of compound 2 and the amino or nitrogen group of the rigid linker, followed by a condensation reaction with the carboxyl or ketone carbonyl group of E3 ligase ligand A with a flexible linker, to obtain the compound of general formula (I); the second reaction is a condensation reaction between the halogen atom of compound 2 and the amino group of E3 ligase ligand B with a flexible linker, to obtain the estrogen receptor protein targeted degradation chimeric derivative of general formula (I) based on the estradiol structure.

[0091] Optionally, the molar ratio of compound 2 to E3 ligase ligand B with a flexible linker is 1:1-1.2.

[0092] Optionally, the acid-binding agent in step (1) is a base, which is selected from at least one of pyridine, triethylamine, diisopropylethylamine, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide, and diisopropylethylamine.

[0093] Optionally, the monomer containing halogen atoms and acyl halide groups in step (1) is selected from at least one of bromoacetyl chloride, chloroacetyl chloride, iodoacetic acid chloride, bromopropionyl chloride, chloropropionyl chloride, iodopropionyl chloride, bromoacetyl bromide, and chloroacetyl bromide.

[0094] Optionally, in step (1), the molar ratio of fulvestrant to monomers containing halogen atoms and acyl halide groups is 1:1-1.3, and the molar ratio of acid-binding agent to fulvestrant is 1:2-3.

[0095] Optionally, the monomer having an amino and a tert-butoxycarbonyl group is selected from at least one of [4,4'-bipiperidine]-1-carboxylic acid tert-butyl ester, 3,9-diazaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester, 2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester, 4-(piperazin-1-ylmethyl)piperazin-1-carboxylic acid tert-butyl ester, and piperazine-1-carboxylic acid tert-butyl ester.

[0096] Optionally, the E3 ligase ligand A with a flexible linker is selected from 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)acetic acid, 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindolline-1,3-dione.

[0097] Optionally, the E3 ligase ligand B with a flexible linker is selected from N-(2-(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)oxy)acetamide.

[0098] Optionally, step (2) involves reacting compound 2 with a monomer containing amino and tert-butoxycarbonyl groups and an E3 ligase ligand A containing a flexible linker to obtain the estrogen receptor protein-targeting degradation chimeric derivative of general formula (I) based on an estradiol structure, specifically as follows:

[0099] (2-1): Compound 2 is condensed with a monomer containing amino and tert-butoxycarbonyl groups to obtain intermediate A.

[0100] (2-2): The protecting group of intermediate A is removed under acidic conditions to obtain intermediate B containing an amino group;

[0101] (2-3): Intermediate B undergoes a condensation reaction with E3 ligase ligand A with a flexible linker to obtain a chimeric derivative of estrogen receptor protein targeting degradation based on the estradiol structure of general formula (I).

[0102] In step (2-1), the halogen atom of compound 2 undergoes a condensation reaction with the amino group of the monomer containing amino and tert-butoxycarbonyl groups to obtain intermediate A. Given the structure of the monomer containing amino and tert-butoxycarbonyl groups listed above, after reacting with compound 2, it has a protecting group at the end. Therefore, in step (2-2), intermediate A is deprotected under acidic conditions to obtain intermediate B containing amino groups. In step (2-3), the amino group of intermediate B undergoes a condensation reaction with the carboxyl or ketone carbonyl group of E3 ligase ligand A containing a flexible linker to obtain the estrogen receptor protein targeted degradation chimeric derivative of general formula (I) based on the estradiol structure.

[0103] In the condensation reactions of steps (2-1) and (2-3) above, a condensing agent and a base are added. The condensing agent is selected from at least one of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, N,N'-diisopropylcarbodiimide, and dicyclohexylcarbodiimide. The base is selected from at least one of triethylamine, potassium carbonate, sodium carbonate, and diisopropylethylamine.

[0104] The acidic environment in step (2-2) is provided by HCl or TFA, and the pH of the acidic environment is 3-5.

[0105] Optionally, in step (2-1), the molar ratio of compound 2 to the monomer with amino and tert-butoxycarbonyl groups is 1:1-1.2, and in step (2-3), the molar ratio of intermediate B to E3 ligase ligand A with flexible linker is 1:1-1.2.

[0106] According to a third aspect of this application, another method for preparing the estrogen receptor protein-targeted degradation chimeric derivative based on the estradiol structure described in any of the preceding claims is provided, comprising the following steps:

[0107] S1: Fluvestrant is stirred with a monomer containing carboxyl and tert-butyloxycarbonyl groups in the presence of a condensing agent and a base to obtain an intermediate containing tert-butyloxycarbonyl groups.

[0108] S2: The intermediate containing tert-butyloxycarbonyl protected group obtained in S1 is placed in an acidic environment to remove the protecting group and obtain an intermediate containing an amino group.

[0109] S3: The amino-containing intermediate obtained in S2 is reacted with E3 ligase ligand A with a flexible linker, and extracted and purified to obtain a chimeric derivative of estrogen receptor protein targeting degradation based on the estradiol structure of general formula (I). The E3 ligase ligand A with a flexible linker has a carboxyl group or a ketone carbonyl group at its end.

[0110] In step S1, the molar ratio of fulvestrant to monomers with carboxyl and tert-butyloxycarbonyl groups and condensing agent is 1:(1.1-1.2):(1.1-1.2).

[0111] In step S1 above, the monomer with carboxyl and tert-butoxycarbonyl groups is selected from at least one of 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid and 1'-(tert-butoxycarbonyl)-[1,4'-bipiperidine]-4-carboxylic acid.

[0112] In step S3 above, the E3 ligase ligand A with a flexible linker is selected from at least one of lenalidomide, thalidomide, 4-hydroxy-thalidomide, 5-hydroxy-thalidomide, pomalidomide, 3-(1-oxoisoindolin-2-yl)piperidin-2,6-dione, 4-amino-4-oxo-2-(1-oxoisoindolin-2-yl)butyric acid, or 2-(1-methyl-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione or its derivatives. A condensing agent is added to the reaction in step S3 and the reaction is carried out at room temperature.

[0113] The condensing agent in steps S1 and S3 is selected from at least one of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, N,N'-diisopropylcarbodiimide, and dicyclohexylcarbodiimide; the base is selected from at least one of triethylamine, potassium carbonate, sodium carbonate, and diisopropylethylamine.

[0114] The acidic environment in step S2 above is provided by HCl or TFA, and the pH of the acidic environment is 3-5.

[0115] According to a fourth aspect of this application, a method for preparing another estrogen receptor protein-targeted degradation chimeric derivative based on the estradiol structure described in any of the preceding claims is provided, comprising the following steps:

[0116] D1: Dissolve fulvestrant and dihaloalkane in a solvent, add an acid-binding agent and stir to obtain FWSQ-C5;

[0117] D2: The compound containing the amino group and the E3 ligase ligand end and FWSQ-C5 were dissolved in an organic solvent, N,N diisopropylethylamine and sodium iodide were added, and after stirring and reaction was completed, the mixture was extracted and purified to obtain the estrogen receptor protein targeted degradation chimeric derivative of general formula (I) based on the estradiol structure.

[0118] Optionally, the dihaloalkane in step D1 is selected from at least one of bromoiodomethane, bromochloromethane, bromochloroethane, bromoiodopropane, bromochloropropane, bromoiodobutane, and bromochlorobutane; the acid-binding agent is selected from at least one of triethylamine, diisopropylethylamine, potassium carbonate, cesium carbonate, and sodium hydroxide.

[0119] Optionally, in step D2, the E3 ligase ligand terminus of the compound containing the amino group and the E3 ligase ligand terminus is selected from at least one of lenalidomide, thalidomide, 4-hydroxy-thalidomide, 5-hydroxy-thalidomide, pomalidomide, 3-(1-oxoisoindolin-2-yl)piperidin-2,6-dione, 4-amino-4-oxo-2-(1-oxoisoindolin-2-yl)butyric acid, or 2-(1-methyl-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione or a derivative thereof.

[0120] Optionally, the compound containing the amino group and the E3 ligase ligand terminus in step D2 is obtained by reacting 2-(2,6-dioxanediol-3-yl)-5-fluoroisoindole-1,3-dione with an amino-containing linker and then removing the amino protecting group. The specific preparation method is as follows:

[0121] Linker monomers containing amino groups and Boc-protected amino groups were mixed with 2-(2,6-dioxanediol-3-yl)-5-fluoroisoindole-1,3-dione, and reacted with an acid-binding agent. After separation, a combination of CRBN and Linker containing Boc protection was obtained.

[0122] The Boc-protected CRBN and Linker complex was then placed in an acidic environment (HCl, TFA) at pH 3-5 to remove the amino protecting group, yielding a compound containing an amino group and an E3 ligase ligand terminus.

[0123] Optionally, the reaction temperature in step D2 is at room temperature, preferably 10-35℃.

[0124] According to a fifth aspect of this application, a pharmaceutical composition is provided comprising: a therapeutically effective amount of any of the above-described estrogen receptor protein-targeting degradation chimeric derivatives based on an estradiol structure or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0125] According to the sixth aspect of this application, the use of the estrogen receptor protein targeted degradation chimeric derivative based on the estradiol structure described in any of the above claims or the above pharmaceutical composition in the preparation of a medicine for the prevention and treatment of breast cancer is provided.

[0126] According to the seventh aspect of this application, the use of the estrogen receptor protein targeted degradation chimeric derivative based on the estradiol structure of any of the above claims or the above pharmaceutical composition in the preparation of a medicament for treating diseases related to estrogen receptor protein targeted degradation chimeric is provided.

[0127] Optionally, the estrogen receptor protein degradation targeting chimeric-related diseases are selected from tumors or cancers.

[0128] Preferably, the estrogen receptor protein degradation targeting chimeric-related diseases are selected from breast cancer, endometrial cancer, ovarian cancer, uterine cancer, prostate cancer, endometriosis, lung cancer, and esophageal cancer.

[0129] More preferably, the estrogen receptor protein degradation targeting chimeric-related diseases are selected from breast cancer.

[0130] The beneficial effects of this application include, but are not limited to:

[0131] 1. The estrogen receptor protein-targeting degradation chimeric derivative based on the estradiol structure of this application not only has a good inhibitory effect on estrogen-induced signal transduction, but also has a good degradation effect on ERα.

[0132] 2. The estrogen receptor protein targeted degradation chimeric derivative based on the estradiol structure of this application has good pharmacokinetic, bioavailability and in vivo efficacy properties, and can be developed into a novel estrogen receptor degradation agent drug.

[0133] 3. The estrogen receptor protein-targeting chimeric derivative based on the estradiol structure of this application has high protein degradation ability and relatively low dosage, which can improve the safety window of drug therapy.

[0134] 4. The estrogen receptor protein targeted degradation chimeric derivative based on the estradiol structure of this application can achieve both oral and injectable administration, thereby achieving greater convenience in drug delivery. Attached Figure Description

[0135] The accompanying drawings, which are included to provide a further understanding of this application and form part of this application, illustrate exemplary embodiments and are used to explain this application, but do not constitute an undue limitation of this application. In the drawings:

[0136] Figure 1 This is a schematic diagram of a Western blotting experiment illustrating the inhibition of ER protein levels in MCF-7 cells by QDE-FWSQ-12, as described in the embodiments of this application.

[0137] Figure 2 This is a schematic diagram of a Western blotting experiment involving QDE-FWSQ-4 inhibiting ER protein levels in MCF-7 cells, as described in the embodiments of this application.

[0138] Figure 3 This is a schematic diagram of a Western blot experiment showing the inhibition of ER protein levels in MCF-7 cells by QDE-FWSQ-5, QDE-FWSQ-6, QDE-FWSQ-11, QDE-FWSQ-12, QDE-FWSQ-13, and QDE-FWSQ-21 involved in the embodiments of this application.

[0139] Figure 4 A WB assay to quantify protein concentrations in ER protein degradation. Detailed Implementation

[0140] The present application is described in detail below with reference to the embodiments, but the present application is not limited to these embodiments.

[0141] Unless otherwise specified, the raw materials used in the embodiments and comparative examples of this application were all purchased commercially.

[0142] Unless otherwise specified, the methods used in the embodiments and comparative examples of this application are conventional methods in the prior art.

[0143] Related definitions

[0144] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0145] The term "pharmaceutically acceptable" as used herein refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0146] The term "pharmaceutically acceptable salt" refers to the salt of the compounds in this application, prepared by reacting a compound with a relatively non-toxic acid or base, as discovered in this application, with a specific substituent. When the compounds in this application contain relatively acidic functional groups, a base addition salt can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. When the compounds in this application contain relatively basic functional groups, an acid addition salt can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Certain specific compounds in this application contain both basic and acidic functional groups, and thus can be converted into either a base or an acid addition salt.

[0147] The pharmaceutically acceptable salts of this application can be synthesized from parent compounds containing an acid radical or a base using conventional chemical methods. Generally, such salts are prepared by reacting these compounds, in their free acid or base form, with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture of both.

[0148] The compounds of this application may exist in specific geometric or stereoisomeric forms. This application envisions all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (2)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this application. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this application.

[0149] The compounds and intermediates of this application may also exist in different tautomer forms, and all such forms are included within the scope of this application. The terms "tautomer" or "tautomer form" refer to structural isomers with different energies that can interconvert via low energy barriers. For example, proton tautomers (also known as proton transfer tautomers) include interconversions via proton migration, such as keto-enol and imine-enamine isomerization. A specific example of a proton tautomer is the imidazole moiety, in which a proton can migrate between two ring nitrogens. Valence tautomers include interconversions via the recombination of some bonding electrons. The compounds of this application may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compound may be labeled with radioactive isotopes such as tritium (2H), iodine-125 (125I), or C-14 (14C). For example, deuterium can be used to replace hydrogen to form deuterated drugs. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic transformations of the compounds in this application, regardless of radioactivity, are included within the scope of this application.

[0150] The terms “optional” or “optionally” refer to events or conditions that may occur but are not required to occur as described below, and the description includes both cases where said events or conditions occur and cases where said events or conditions do not occur.

[0151] The term "targeting chimera" refers to a bifunctional molecule containing two small molecule ligands: one with high affinity for the target protein and a second for recruiting an E3 ligase that ubiquitinates the protein and targets it for proteolysis via the proteasome.

[0152] The compounds of this application can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of this application.

[0153] The solvents used in this application are commercially available. Compounds are named according to conventional naming practices in the art; commercially available compounds are named using supplier directory names.

[0154] Example 1

[0155] This embodiment relates to a method for preparing an estrogen receptor protein-targeted degradation chimeric derivative of general formula (I) based on an estradiol structure, comprising the following steps:

[0156] (1) Place fulvestrant in a solvent, add an acid-binding agent and lower the temperature to 1-5℃, then add a monomer containing halogen atoms and acyl halide groups to react and obtain compound 2;

[0157] (2) Compound 2 is reacted sequentially with a rigid linker and an E3 ligase ligand A with a flexible linker to obtain an estrogen receptor protein-targeting degradation chimeric derivative of general formula (I) based on an estradiol structure. The E3 ligase ligand A with a flexible linker has a carboxyl group or a ketone carbonyl group at its end; or

[0158] Compound 2 was reacted with E3 ligase ligand B with a flexible linker to obtain an estrogen receptor protein targeted degradation chimeric derivative of general formula (I) based on an estradiol structure, wherein the E3 ligase ligand B with the flexible linker has an amino group at its end.

[0159] Preferably, step (2) involves reacting compound 2 with a monomer containing amino and tert-butoxycarbonyl groups and an E3 ligase ligand A containing a flexible linker in sequence to obtain the estrogen receptor protein-targeting degradation chimeric derivative of general formula (I) based on an estradiol structure, specifically as follows:

[0160] (2-1): Compound 2 is condensed with a rigid linker to obtain intermediate A;

[0161] (2-2): The protecting group of intermediate A is removed under acidic conditions to obtain intermediate B containing an amino group;

[0162] (2-3): Intermediate B undergoes a condensation reaction with E3 ligase ligand A with a flexible linker to obtain a chimeric derivative of estrogen receptor protein targeting degradation based on the estradiol structure of general formula (I).

[0163] The following estrogen receptor protein-targeted degradation chimeric derivatives based on estradiol structures were prepared according to the above preparation method, as follows:

[0164] Compound QDE-FWSQ-1

[0165] The specific steps and reaction equations are as follows:

[0166]

[0167] (1) Dissolve 121.4 g of fluoroviscon (0.2 mmol) in dichloromethane (5 ml), add 40.4 mg of triethylamine (0.4 mmol), cool to 3 °C, add 31.5 mg of bromoacetyl chloride (0.2 mmol) dropwise, purge with nitrogen three times, react at 3 °C for 4 hours, monitor the reaction progress with TLC, quench the reaction with saturated ammonium chloride aqueous solution after the reaction is complete, extract three times with dichloromethane, and separate the dichloroacetyl chloride using a separatory funnel. The methane layer was evaporated to dryness under vacuum and purified by column chromatography to give 112.4 mg of (7R,8S,9S,12R,14S,17S)-17-hydroxy-12-methyl-7-(9-((4,4,5,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopentan[a]phenanthrene-3-yl-2-bromoacetate, in 77.2% yield, i.e., compound 2. The mass of this compound was accumulated through multiple reactions.

[0168] (2-1) 112.4 mg of compound 2 (0.154 mmol) and 45.4 mg of [4,4'-bipiperidine]-1-carboxylic acid tert-butyl ester (0.169 mmol) were dissolved in 3 mL of dichloromethane. 46.75 mg of potassium carbonate (0.338 mmol) was added, and the reaction was carried out at 40 °C for 6 h. The reaction progress was monitored by TLC. After the reaction was complete, water / dichloromethane was added for extraction. The dichloromethane phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel preparative chromatography to obtain 100.3 mg. 1'-(2-(((7R,8S,9S,12R,14S,17S)-17-hydroxy-12-methyl-7-(9-(4,4,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopentan[a]phenanthrene-3-yl)oxy)-2-oxoethyl)-[4,4'-bipiperidine]-1-carboxylic acid tert-butyl ester, yield 71.3%, namely compound FWSQ1-1.

[0169] (2-2) Dissolve 110.3 mg FWSQ1-1 (0.116 mmol) in 5 ml of dichloromethane, cool to 0 °C in an ice bath, and slowly add 0.5 ml of trifluoroacetic acid dropwise to the reaction system. After the addition is complete, remove the ice bath and react at room temperature for 3 h. Quench the reaction with sodium bicarbonate solution, extract with dichloromethane and water three times, separate the dichloromethane layer with a separatory funnel, and purify by column chromatography at 30 °C to obtain (7R, 8S, 9S). 87.1 mg of compound FWSQ1-2 was obtained from 12R,14S,17S)-17-hydroxy-12-methyl-7-(9-((4,4,5,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopentan[a]phenanthrene-3-yl-2-([4,4'-bipiperidine]-1-yl)acetate, with a yield of 91.5%.

[0170] (2-3) Add 35.5 mg of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)acetic acid (0.107 mmol) to a 10 ml single-necked flask, add 2 ml of DCM for suspension, add 40.7 mg (0.107 mmol) of HATU (2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) and 27.6 mg (0.214 mmol) of diisopropylethylamine, stir and activate for 30 min, then add 87.1 mg of compound FWSQ-1-2 (0.107 mmol). The reaction was carried out at room temperature for 3 hours, and the reaction progress was monitored by TLC. After the reaction was completed, the mixture was extracted with ethyl acetate and water. After three extractions, the ethyl acetate was separated by a separatory funnel, and purified by vacuum distillation and column chromatography to obtain 53.3 mg of (7R,8S,8S,9S,12R,14S,17S)-17-hydroxy-12-methyl-7-(5,4,4,5,5,5,5-5-pentafluoroformyl9,9,11,11,12,13,14,15,16,16,17,17-decahydro-6H-cyclopentene,4'-piperidin]-1-yl) acetate, with a yield of 44.2%, namely compound QDE-FWSQ-1.

[0171] The structure of QDE-FWSQ-1 was confirmed by mass spectrometry (HR-ESI-MS) and proton nuclear magnetic resonance (1H-NMR): ESI-MS (positive mode) m / z: 1158.5 [M+H] + (calculated for C) 60 H 80 F5N5O 10S, 1157.5). 1H-NMR (600Hz, CDCl3): 11.033 (NH, 1H), 8.012 (Ph-CH, 1H), 7.677 (Ph-H, 2H), 6.795-6.933 (Ph-H, 3H), 5.53 (O-CH2, 2H), 4.56 (N-CH2, 2H), 4.769 ( C=O-CH2,2H), 4.428 ( N-CH, 1H), 4.134 ( -OH, 1H), 3.416-3.553(CH2, 7H), 2.349-2.909(CH2, CH, 11H), 1.266-2.243(CH2, CH, 43H). 0.893 (CH3, 3H).

[0172] Compound QDE-FWSQ-5

[0173] The specific steps and reaction equations are as follows:

[0174]

[0175] (1) Same as compound QDE-FWSQ-1.

[0176] (2-1) 112.4 mg of compound 2 (0.154 mmol) and 42.9 mg of 3,9-diazaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (0.169 mmol) were dissolved in 3 mL of dichloromethane. 46.75 mg of potassium carbonate (0.338 mmol) was added, and the reaction was carried out at 40 °C for 6 h. The reaction progress was monitored by TLC. After the reaction was completed, water / dichloromethane was added for extraction. The dichloromethane phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel preparative plate to obtain 115.5 mg. 9-(2-(((7R,8R,9R,13S,14S,17S)-17-hydroxy-8,9,13,14-tetramethyl-7-(9-(4,4,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopentan[a]phenanthrene-3-yl)oxy)-2-oxoethyl)-3,9-diazaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester, yield 79.6%, i.e., compound 5-1.

[0177] (2-2) 115.5 mg of compound 5-1 (0.122 mmol) was dissolved in 5 mL of dichloromethane. The mixture was cooled to 0 °C in an ice bath. 0.5 mL of trifluoroacetic acid was slowly added dropwise to the reaction system. After the addition was complete, the ice bath was removed, and the reaction was allowed to proceed at room temperature for 3 h. The reaction was quenched with sodium bicarbonate solution, and the mixture was extracted three times with dichloromethane and water. The dichloromethane layer was separated using a separatory funnel, and purified by column chromatography at 30 °C under reduced pressure to obtain 93.5 mg of (7R, 8R, 9R, 13S, 14S) compounds. ,17S)-17-hydroxy-8,9,13,14-tetramethyl-7-(9-(4,4,4,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopentan[a]phenanthrene-3-yl-2-(3,9-diazaspiro[5.5]undecane-3-yl)acetate, yield 90.6%, i.e. compound 5-2.

[0178] (2-3) 36.5 mg of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)acetic acid (same as in Example 1) (0.110 mmol) was added to a 10 ml single-necked flask, 2 ml of DCM was added for suspension, 41.8 mg (0.110 mmol) of HATU (same as in Example 1) and 28.3 mg (0.220 mmol) of diisopropylethylamine (same as in Example 1) were added and stirred for 30 min to activate the mixture. 93.5 mg of compound 5-2 (0.110 mol) was added and the mixture was reacted at room temperature for 3 h. The reaction progress was monitored by TLC. After the reaction was completed, the mixture was filtered. Extracted with ethyl acetate and water three times, the ethyl acetate was separated by a separatory funnel, and purified by vacuum distillation and column chromatography to obtain 52.5 mg of (7R,8R,9R,13S,14S,17S)-17-hydroxy-8,9,13,14-tetramethyl-7-(9-(4,4,4,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12, 13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3-yl-2-(9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxaindol-4-yl)oxy)acetyl)-3,9-diazaspiro[5.5]undecane-3-yl)acetate, yielding 41.3%, namely compound QDE-FWSQ-5.

[0179] The structure of QDE-FWS-5 was confirmed by mass spectrometry (HR-ESI-MS) and proton nuclear magnetic resonance (1H-NMR): ESI-MS (positive mode) m / z: 1115.5 [M+H] + (calculated for C) 58 H 75 F5N4O 10S, 1114.5594). 1H-NMR (600Hz, CDCl3): 11.323(NH, 1H), 7.9137 (Ph-CH, 1H), 7.594 (Ph-H 2H), 6.764-6.964 (Ph-H, 3H), 4.994 (C=O-CH2, 2H), 4.754 (-OH, 1H), 4.483 (N-CH,1H), 3.782 (N-CH2, 4H), 3.572 (N-CH2, 2H), 3.4134 (OH-CH, 1H), 2.832-2.922(Ph-CH2, 2H), 2.433 - 2.594 (CH2, CH, 10H), 1.902-2.233(CH2,4H), 1.254-1.889(CH2,38H), 0.855(CH3, 3H).

[0180] Compound QDE-FWSQ-6

[0181] The specific preparation steps and reaction equations are as follows:

[0182]

[0183] (1) Same as compound QDE-FWSQ-1.

[0184] (2-1) 112.4 mg of compound 2 (0.154 mmol) and 40.59 mg of 2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (0.169 mmol) were dissolved in 3 ml of dichloromethane. 46.75 mg of potassium carbonate (0.338 mmol) was added, and the mixture was reacted at 40 °C for 6 h. The reaction progress was monitored by TLC. After the reaction was complete, water / dichloromethane was added for extraction. The dichloromethane phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel preparative chromatography to obtain 100.2 mg of tert-butyl2-(2-(((7R,8R,9R,13S,14S,17S)-17-hydroxy-8,9,13,14-tetramethyl-7-(9-((4,4,5,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13, 14,15,16,17-Decahydro-6H-cyclopentan[a]phenanthrene[3,2-c]azapyro-3-yl)oxy)-2-oxoethyl)-2,8-dithioneheterocyclic[4,5]octane-8-carboxylate, yield 70.1%, i.e., compound 6-1

[0185] (2-2) 100.2 mg of compound 6-1 (0.108 mmol) was dissolved in 5 ml of dichloromethane. The mixture was cooled to 0 °C in an ice bath. 0.5 ml of trifluoroacetic acid was slowly added dropwise to the reaction system. After the addition was complete, the ice bath was removed, and the reaction was allowed to proceed at room temperature for 3 h. The reaction was then quenched with sodium bicarbonate solution. The mixture was extracted three times with dichloromethane and water. The dichloromethane layer was separated using a separatory funnel and purified by column chromatography under reduced pressure at 30 °C to obtain 79.0 mg (7R, 8R, 9...). R, 13S, 14S, 17S)-17-hydroxy-8,9,13,14-tetramethyl-7-(9-(4,4,4,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3-yl-2-(3,9-diazaspiro[5.5]undecane-3-yl)acetate, yield 88.3%, i.e., compound 6-2.

[0186] (2-3) 31.6 mg of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)acetic acid (0.095 mmol) was added to a 10 ml single-necked flask, 2 ml of DCM was added for suspension, 36.1 mg (0.095 mmol) of HATU and 24.5 mg (0.19 mmol) of diisopropylethylamine were added and stirred for 30 min to activate the mixture. 79.0 mg of compound 6-2 (0.095 mol) was added and the mixture was reacted at room temperature for 3 h. The reaction progress was monitored by TLC. After the reaction was completed, the mixture was filtered. Extracted with ethyl acetate and water three times, the ethyl acetate was separated by a separatory funnel, purified by vacuum distillation and column chromatography to obtain 42.6 mg of (7R,8R,9R,13S,14S,17S)-17-hydroxy-8,9,13,14-tetramethyl-7-(9-((4,4,5,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3-yl 2-(8-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)oxy)acetyl)-2,8-diazaspiro[4.5]decane-2-yl)acetate, with a yield of 39.3%, namely compound QDE-FWSQ-6.

[0187] The structure of QDE-FWS-6 was confirmed by mass spectrometry (HR-ESI-MS) and proton nuclear magnetic resonance (1H-NMR): ESI-MS (positive mode) m / z: 1111.5 [M+H] + (calculated for C) 57 H 73 F5N4O 10S, 1110.4968). 1H-NMR (600Hz, CDCl3): 11.323(NH, 1H), 7.9137 (Ph-CH, 1H), 7.594 (Ph-H 2H), 6.764-6.964 (Ph-H, 3H), 4.994 (C=O-CH2, 2H), 4.754 (-OH, 1H), 4.483 (N-CH,1H), 3.782 (N-CH2, 4H), 3.572 (N-CH2, 2H), 3.4134 (OH-CH, 1H), 2.832-2.922(Ph-CH2, 2H), 2.433-2.594(CH2, 9H), 1.764-1.944(CH2, 6H), 1.902-2.233(CH2CH,5H) 1.254-1.794(CH2, 30H), 0.855(CH3, 3H).

[0188] Compound QDE-FWSQ-11

[0189] The specific preparation steps and reaction equations are as follows:

[0190]

[0191] (1) Same as compound QDE-FWSQ-1.

[0192] (2-1) 112.4 mg of compound 2 (0.154 mmol) and 48.0 mg of 4-(piperazin-1-ylmethyl)piperazin-1-carboxylic acid tert-butyl ester (0.169 mmol) were dissolved in 3 mL of dichloromethane. 46.75 mg of potassium carbonate (0.338 mmol) was added, and the reaction was carried out at 40 °C for 6 h. The reaction progress was monitored by TLC. After the reaction was complete, water / dichloromethane was added for extraction. The dichloromethane phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel preparative plate to obtain 133.7 mg. 4-(4-(2-(((7R,8R,9R,13S,14S,17S)-17-hydroxy-8,9,13,14-tetramethyl-7-(9-(4,4,4,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3-yl)oxy)-2-oxoethyl)piperazin-1-yl)methyl)piperazin-1-carboxylic acid ester, yield 81.4%, i.e., compound 11-1.

[0193] (2-2) 133.7 mg of compound 11-1 (0.137 mmol) was dissolved in 5 mL of dichloromethane. The mixture was cooled to 0 °C in an ice bath. 0.5 mL of trifluoroacetic acid was slowly added dropwise to the reaction system. After the addition was complete, the ice bath was removed, and the reaction was allowed to proceed at room temperature for 3 h. The reaction was then quenched with sodium bicarbonate solution. The mixture was extracted three times with dichloromethane and water. The dichloromethane layer was separated using a separatory funnel, and purified by column chromatography under reduced pressure at 30 °C to obtain 101.9 mg of the compound. (7R,8R,9R,13S,14S,17S)-17-hydroxy-8,9,13,14-tetramethyl-7-(9-(4,4,4,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3-yl-2-(4-(piperazin-1-ylmethyl)piperazin-1-yl)acetate, yield 91.4%, i.e., compound 11-2.

[0194] (2-3) Add 41.5 mg of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)acetic acid (0.125 mmol) to a 10 ml single-necked flask, add 2 ml of DCM suspension, add 47.6 mg (0.125 mmol) of HATU and 32.3 mg (0.25 mmol) of diisopropylethylamine, stir and activate for 30 min, add 101.9 mg of compound 11-2 (0.125 mol), react at room temperature for 3 h, monitor the reaction progress by TLC, and filter after the reaction is complete. Extracted with ethyl acetate and water three times, the ethyl acetate was separated by a separatory funnel, purified by vacuum distillation and column chromatography to give 65.6 mg of (7R,8R,9R,13S,14S,17S)-17-hydroxy-8,9,13,14-tetramethyl-7-(9-(4,4,4,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3-yl-2-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)acetyl)piperazin-1-yl)methyl)piperazin-1-alkyl)acetate, in 44.2% yield, namely compound QDE-FWSQ-11.

[0195] The structure of QDE-FWS-11 was confirmed by mass spectrometry (HR-ESI-MS) and proton nuclear magnetic resonance (1H-NMR): ESI-MS (positive mode) m / z: 1145.5 [M+H] + (calculated for C) 58 H 77 F5N6O 10S, 1144.5437). 1HNMR: δ 7.86 (Ph-H, 1H), 7.24 (Ph-H, 1H), 7.00 (Ph-H, 1H), 6.91 (Ph -H, 1H), 6.89 (Ph -H, 1H), 6.82 (Ph -H, 1H), 5.53 (Ph -H, 1H), 5.37 (Ph -H ,1H), 5.07(Ph, 1H), 4.36 (CO-CH2, 2H), 4.24 (CO-CH 2, 2H), 3.65 (OH, 1H), 3.48 (N-CH2,2H), 3.37 (N-CH2, 4H), 3.30 (N-CH2, 4H), 3.09 (OH-CH, 1H), 3.01 (CH2, 8H),2.84 (CH2, 2H), 2.57 (CH2, 4H), 2.24 (CH2, 2H), 1.76-1.78 (CH2, 6H), 1.64-1.67(CH2, 4H), 1.25-1.55 (CH2 / CH, 22H), 0.91 (CH, 1H), 0.80 (CH3, 3H).

[0196] Compound QDE-FWSQ-12

[0197] The specific preparation steps and reaction equations are as follows:

[0198]

[0199] (1) Same as compound QDE-FWSQ-1.

[0200] (2-1) 112.4 mg of compound 2 (0.154 mmol) and 31.4 mg of piperazine-1-carboxylic acid tert-butyl ester (0.169 mmol) were dissolved in 3 mL of dichloromethane. 46.75 mg of potassium carbonate (0.338 mmol) was added, and the mixture was reacted at 40 °C for 6 h. The reaction progress was monitored by TLC. After the reaction was complete, the mixture was extracted with water / dichloromethane, dried over anhydrous sodium sulfate in the dichloromethane phase, filtered, concentrated, and purified by silica gel preparative chromatography to obtain 98.8 mg. 4-(2-(((7R,8R,9R,13S,14S,17S)-17-hydroxy-8,9,13,14-tetramethyl-7-(9-(4,4,5,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3-yl)oxy)-2-oxoethyl)piperazine-1-carboxylic acid tert-butyl ester, yield 73.4%, i.e., compound 12-1.

[0201] (2-2) 98.8 mg of compound 12-1 (0.113 mmol) was dissolved in 5 ml of dichloromethane. The mixture was cooled to 0 °C in an ice bath. 0.5 ml of trifluoroacetic acid was slowly added dropwise to the reaction system. After the addition was complete, the ice bath was removed, and the reaction was allowed to proceed at room temperature for 3 h. The reaction was then quenched with sodium bicarbonate solution. The mixture was extracted three times with dichloromethane and water. The dichloromethane layer was separated using a separatory funnel and purified by column chromatography under reduced pressure at 30 °C to obtain 80.7 mg (7R, 8R, 9R, 13S, 14S). 17S)-17-hydroxy-8,9,13,14-tetramethyl-7-(9-(4,4,4,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopentan[a]phenanthrene-3-yl-2-(3,9-diazaspiro[5.5]undecane-3-yl)acetate, in 92.3% yield, i.e., compound 12-2.

[0202] (2-3) 34.5 mg of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)acetic acid (0.104 mmol) was added to a 10 ml single-necked flask, 2 ml of DCM was added for suspension, 39.5 mg (0.104 mmol) of HATU and 26.8 mg (0.208 mmol) of diisopropylethylamine were added and stirred for 30 min to activate the mixture. 80.7 mg of compound 12-2 (0.104 mol) was added and the mixture was reacted at room temperature for 3 h. The reaction progress was monitored by TLC. After the reaction was completed, the mixture was filtered. Extracted with ethyl acetate and water three times, the ethyl acetate was separated by a separatory funnel, purified by vacuum distillation and column chromatography to give 42.6 mg of (7R,8S,9S,12R,14S,17S)-17-hydroxy-12-methyl-7-(9-((4,4,5,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopentan[a]phenanthrene-3-yl-2-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)acetyl)piperazin-1-yl)acetate, in 39.2% yield, namely compound QDE-FWS-12.

[0203] The structure of QDE-FWS-12 was confirmed by mass spectrometry (HR-ESI-MS) and proton nuclear magnetic resonance (1H-NMR): ESI-MS (positive mode) m / z: 1047.4 [M+H] + (calculated for C) 53 H 67 F5N4O 10S, 1046.4498). 1H-NMR (600Hz, CDCl3): 11.323(NH, 1H), 7.9137 (Ph-CH, 1H), 7.594 (Ph-H 2H), 6.764-6.964 (Ph-H, 3H), 4.994 (C=O-CH2, 2H), 4.754 (-OH, 1H), 4.483 (N-CH,1H), 3.782 (N-CH2, 4H), 3.572 (N-CH2, 2H), 3.4134 (OH-CH, 1H), 2.690-2.922(CH2,CH, 11H), 2.433-2.594(CH2, 4H), 2.093-2.233(CH2, 4H) 1.254-1.994(CH2,26H), 0.855(CH3,CH, 4H).

[0204] Compound QDE-FWSQ-13

[0205] The specific preparation steps and reaction equations are as follows:

[0206]

[0207] (1) Same as compound QDE-FWSQ-1.

[0208] (2-1) 112.4 mg of compound 2 (0.154 mmol) and 42.9 mg of 3,9-diazaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (0.169 mmol) were dissolved in 3 mL of dichloromethane. 46.75 mg of potassium carbonate (0.338 mmol) was added, and the reaction was carried out at 40 °C for 6 h. The reaction progress was monitored by TLC. After the reaction was completed, water / dichloromethane was added for extraction. The dichloromethane phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel plate preparation to obtain 115.5 mg. 9-(2-(((7R,8R,9R,13S,14S,17S)-17-hydroxy-8,9,13,14-tetramethyl-7-(9-(4,4,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopentan[a]phenanthrene-3-yl)oxy)-2-oxoethyl)-3,9-diazaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester, yield 79.6%, i.e., compound 13-1.

[0209] (2-2) 115.5 mg of compound 13-1 (0.122 mmol) was dissolved in 5 ml of dichloromethane. The mixture was cooled to 0 °C in an ice bath. 0.5 ml of trifluoroacetic acid was slowly added dropwise to the reaction system. After the addition was complete, the ice bath was removed, and the reaction was allowed to proceed at room temperature for 3 h. The reaction was then quenched with sodium bicarbonate solution. The mixture was extracted three times with dichloromethane and water. The dichloromethane layer was separated using a separatory funnel and purified by column chromatography under reduced pressure at 30 °C to obtain 93.5 mg of (7R, 8R, 9R, 13S, 14S) compound. 17S)-17-hydroxy-8,9,13,14-tetramethyl-7-(9-(4,4,4,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopentan[a]phenanthrene-3-yl-2-(3,9-diazaspiro[5.5]undecane-3-yl)acetate, in 90.6% yield, i.e., compound 13-2.

[0210] (2-3) 93.5 mg of compound 13-2 (0.110 mol) was added to a 10 ml single-necked flask, 2 ml of DMF was added to dissolve it, 28.3 mg (0.220 mmol) of diisopropylethylamine was added and stirred for 5 min, 30.4 mg of 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (0.110 mmol) was added and the mixture was reacted at 70 °C for 12 h. The reaction progress was monitored by TLC. After the reaction was completed, the mixture was filtered. Extracted with ethyl acetate and water three times, the ethyl acetate was separated by a separatory funnel, and purified by vacuum distillation and column chromatography to obtain 80.5 mg of (7R, 8R, 9S, 13S, 14S, 17S)-17-hydroxy-13-methyl-7-(9-((4,4,5,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopentan[a]phenanthrene-3-yl-2-(9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxaindol-5-yl)-3,9-diazaspiro[5.5]undecane-3-yl)acetate, yield 69.3%, namely compound QDE-FWSQ-13.

[0211] The structure of QDE-FWS-13 was confirmed by mass spectrometry (HR-ESI-MS) and proton nuclear magnetic resonance (1H-NMR): ESI-MS (positive mode) m / z: 1057.5 [M+H] + (calculated for C) 56 H 73F5N4O8S, 1056.5069). 1H-NMR (600Hz, CDCl3): 11.323(NH, 1H), 7.9137 (Ph-CH, 1H), 7.594 (Ph-H 2H), 6.964 (Ph-H, 2H) , 6.764 (Ph-H, 1H) , 4.754 (-OH, 1H), 4.483 (N-CH, 1H), 3.782 (N-CH2, 4H), 3.572 (N-CH2, 2H), 3.4134 (OH-CH, 1H), 2.832-2.922(Ph-CH2, 2H), 2.433 - 2.594 (CH2, CH, 9H), 1.902-2.233(CH2,4H), 1.254-1.889(CH2,39H), 0.855(CH3,3H).

[0212] Compound QDE-FWSQ-21

[0213] The specific preparation steps and reaction equations are as follows:

[0214]

[0215] (1) Same as compound QDE-FWSQ-1.

[0216] (2) 112.4 mg of compound 2 (0.154 mmol) and 85.5 mg of N-(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)oxy)acetamide (0.169 mmol) were dissolved in 3 ml of acetonitrile. 46.75 mg of potassium carbonate (0.338 mmol) was added, and the reaction was carried out at 40 °C for 6 h. The reaction progress was monitored by TLC. After the reaction was completed, water / dichloromethane was added for extraction, the dichloromethane phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel preparative plate to obtain 111.1 mg. (7R,8R,9R,13S,14S,17S)-17-hydroxy-8,9,13,14-tetramethyl-7-(9-(4,4,4,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3-yl(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxaisoindol-4-yl)oxy)-2-oxo-6,9,12-trioxa-3-azatetradecane-14-yl)glycine ester, yield 60.4%, i.e., compound QDE-FWS-21.

[0217] The structure of QDE-FWS-21 was confirmed by mass spectrometry (HR-ESI-MS) and proton nuclear magnetic resonance (1H-NMR): ESI-MS (positive mode) m / z: 1153.5 [M+H] + (calculated for C) 57 H 77 F5N4O 13 S, 1152.5128). 1H-NMR (600Hz, CDCl3): 10.933(NH, 1H), 8.021 (Ph-CH, 1H), 7.502 (Ph-H 2H), 6.839-6.902 (Ph -H, 3H), 5.426 (NH-H, 1H), 4.994 (C=O-CH2, 2H), 4.701 (-OH,1H), 4.483 (N-CH, O-CH23H), 3.782 (NH-CH2, 2H), 3.782 (O-CH2, 2H), 3.572 (O-CH2, 10H), 3.402 (OH-CH, 1H), 3.245 (NH-CH2, 2H), 2.6334-2.722(Ph-CH2,NH-CH,3H), 2.433- 2.594 (S=O -CH2, 4H), 1.764-2.292(CH2,CH, 5H), 1.254-1.794(CH2,CH,31H), 0.855(CH3, 3H).

[0218] Example 2

[0219] This embodiment relates to a method for preparing an estrogen receptor protein-targeted degradation chimeric derivative of general formula (I) based on an estradiol structure, comprising the following steps:

[0220] S1: Fluvestrant is stirred with a monomer containing carboxyl and tert-butyloxycarbonyl groups in the presence of a condensing agent and a base to obtain an intermediate containing tert-butyloxycarbonyl protection.

[0221] S2: The intermediate containing tert-butyloxycarbonyl protected group obtained in S1 is placed in an acidic environment to remove the protecting group and obtain an intermediate containing an amino group.

[0222] S3: The amino-containing intermediate obtained in S2 is reacted with E3 ligase ligand A with a flexible linker, and extracted and purified to obtain a chimeric derivative of estrogen receptor protein targeting degradation based on the estradiol structure of general formula (I). The E3 ligase ligand A with a flexible linker has a carboxyl group or a ketone carbonyl group at its end.

[0223] In step S1, the molar ratio of fulvestrant to monomers with carboxyl and tert-butyloxycarbonyl groups and condensing agent is 1:(1.1-1.2):(1.1-1.2).

[0224] In step S1 above, the monomer with carboxyl and tert-butoxycarbonyl groups is selected from at least one of 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid and 1'-(tert-butoxycarbonyl)-[1,4'-bipiperidine]-4-carboxylic acid.

[0225] In step S3 above, the E3 ligase ligand A with a flexible linker is selected from at least one of lenalidomide, thalidomide, 4-hydroxy-thalidomide, 5-hydroxy-thalidomide, pomalidomide, 3-(1-oxoisoindolin-2-yl)piperidin-2,6-dione, 4-amino-4-oxo-2-(1-oxoisoindolin-2-yl)butyric acid, or 2-(1-methyl-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione, or a derivative thereof. A condensing agent is added to the reaction in step S3, and the reaction is carried out at room temperature.

[0226] The condensing agent in steps S1 and S3 is selected from at least one of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, N,N'-diisopropylcarbodiimide, and dicyclohexylcarbodiimide; the base is selected from at least one of triethylamine, potassium carbonate, sodium carbonate, and diisopropylethylamine.

[0227] The acidic environment in step S2 above is provided by HCl or TFA, and the pH of the acidic environment is 3-5.

[0228] The following estrogen receptor protein-targeted degradation chimeric derivatives based on estradiol structures were prepared according to the above preparation method, as follows:

[0229] Compound QDE-FWSQ-3

[0230] The specific steps and reaction equations are as follows:

[0231]

[0232] S1: 121.4 g of fulvesyl (0.2 mmol) was dissolved in dichloromethane (5 ml), followed by the addition of 50.38 mg of 1-(tert-butyloxycarbonyl)piperidine-4-carboxylic acid (0.22 mmol), 40.4 mg of triethylamine (0.4 mmol), and 45.4 mg of dicyclohexylcarbodiimide (0.22 mmol). The mixture was stirred at room temperature for 24 h, and the reaction progress was monitored by TLC. After the reaction was completed, saturated ammonium chloride aqueous solution was added to quench the reaction. The mixture was extracted three times with dichloromethane, and the dichloromethane layer was separated using a separatory funnel, evaporated to dryness under vacuum, and purified by column chromatography to obtain 78.4 mg. 1-(tert-butyl)4-((7R,8S,9S,12R,14S,17S)-17-hydroxy-12-methyl-7-(9-(4,4,5,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopentan[a]phenanthrene-3-yl)piperidine-1,4-dicarboxylate, i.e., compound 3-1 (0.096 mmol), yield 48.1%.

[0233] S2: Dissolve 78.4 mg of compound 3-1 (0.096 mmol) in 5 mL of dichloromethane. Cool to 0 °C in an ice bath. Slowly add 0.5 mL of trifluoroacetic acid dropwise to the reaction system. After the addition is complete, remove the ice bath and react at room temperature for 3 h. Monitor the reaction progress by TLC. After the reaction is complete, quench with sodium bicarbonate solution. Extract three times with dichloromethane and water. Separate the dichloromethane layer using a separatory funnel. Purify by column chromatography under reduced pressure at 30 °C to obtain the desired product. 62.7 mg (7R,8S,9S,12R,14S,17S)-17-hydroxy-12-methyl-7-(9-((4,4,5,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3-ylpiperidine-4-carboxylic acid ester (0.087 mmol), yield 91.1%, i.e., compound 3-2.

[0234] S3: 28.8 mg of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)acetic acid (0.087 mmol) was added to a 10 ml single-necked flask, 2 ml of DCM was added for suspension, 33.1 mg (0.087 mmol) of HATU and 22.4 mg (0.174 mmol) of diisopropylethylamine were added and the mixture was stirred and activated for 30 min. Then, 62.7 mg of compound 3-2 (0.087 mmol) was added. The reaction was carried out at room temperature for 3 hours, and the reaction progress was monitored by TLC. After the reaction was completed, the mixture was extracted with ethyl acetate and water. After three extractions, ethyl acetate was separated by a separatory funnel, and purified by vacuum distillation and column chromatography to obtain 27.9 mg of (7R,8S,9S,12R,14S,17S)-17-hydroxy-12-methyl-7-(9-((4,4,5,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3-yl-1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)acetyl)piperidin-4-carboxylic acid ester, with a yield of 31.2%, namely compound QDE-FWSQ-3.

[0235] The structure of QDE-FWS-3 was confirmed by mass spectrometry (HR-ESI-MS) and proton nuclear magnetic resonance (1H-NMR): ESI-MS (positive mode) m / z: 1032.4 [M+H]+ (calculated for C 53 H 66 F5N3O 10 ( C=O-CH2, 2H), 4.354 (N-CH, 1H),4.022 (-OH, 1H), 3.416-3.553(N-CH2,OH-H, 5H), 2.549-2.909(CH2, CH, 7H),1.266-2.243(CH2, CH, 40H),0.893(CH3, 3H).

[0236] Compound QDE-FWSQ-4

[0237] The specific steps and reaction equations are as follows:

[0238]

[0239] S1: 121.4 g of fulvesin (0.2 mmol) was dissolved in dichloromethane (5 ml), followed by the addition of 68.64 mg of 1'-(tert-butyloxycarbonyl)-[1,4'-bipiperidine]-4-carboxylic acid (0.22 mmol), 40.4 mg of triethylamine (0.4 mmol), and 45.4 mg of dicyclohexylcarbodiimide (0.22 mmol). The mixture was stirred at room temperature for 24 h, and the reaction progress was monitored by TLC. After the reaction was completed, saturated ammonium chloride aqueous solution was added to quench the reaction. The mixture was extracted three times with dichloromethane, and the dichloromethane layer was separated using a separatory funnel, evaporated to dryness under vacuum, and purified by column chromatography to obtain 108.9 mg. 1'-(tert-butyl)4-((7R,8S,9S,12R,14S,17S)-17-hydroxy-12-methyl-7-(9-(4,4,5,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopentan[a]phenanthrene-3-yl)[1,4'-bipiperidine]-1',4-dicarboxylic acid ester (0.121 mmol), i.e., compound 4-1, yield 60.5%.

[0240] S2: Dissolve 108.9 mg of compound 4-1 (0.121 mmol) in 5 mL of dichloromethane. Cool to 0 °C in an ice bath. Slowly add 0.5 mL of trifluoroacetic acid dropwise to the reaction system. After the addition is complete, remove the ice bath and react at room temperature for 3 h. Monitor the reaction progress by TLC. After the reaction is complete, quench with sodium bicarbonate solution. Extract three times with dichloromethane and water. Separate the dichloromethane layer using a separatory funnel. Purify by column chromatography under reduced pressure at 30 °C to obtain 86... 0.4 mg (7R,8S,9S,12R,14S,17S)-17-hydroxy-12-methyl-7-(9-((4,4,5,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3-yl[1,4'-bipiperidine]-4-carboxylic acid ester (0.108 mmol), yield 89.9%, i.e., compound 4-2.

[0241] S3: Add 34.7 mg of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)acetic acid (0.108 mmol) to a 10 ml single-necked flask, add 2 ml of DCM suspension, add 41.2 mg (0.108 mmol) of HATU and 27.9 mg (0.216 mmol) of diisopropylethylamine, stir and activate for 30 min, then add 86.4 mg of compound 4-2 (0.108 mmol). The reaction was carried out at room temperature for 3 hours, and the reaction progress was monitored by TLC. After the reaction was completed, the mixture was extracted with ethyl acetate and water. After three extractions, ethyl acetate was separated by a separatory funnel, and purified by vacuum distillation and column chromatography to obtain 43.8 mg of (7R,8S,9S,12R,14S,17S)-17-hydroxy-12-methyl-7-(9-((4,4,5,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3-yl 1'-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)acetyl)-[1,4'-bipiperidin]-4-carboxylic acid ester, with a yield of 36.4%, namely compound QDE-FWSQ-4.

[0242] The structure of QDE-FWS-4 was confirmed by mass spectrometry (HR-ESI-MS) and proton nuclear magnetic resonance (1H-NMR): ESI-MS (positive mode) m / z: 1115.5 [M+H]+ (calculated for C 58 H 75 F5N4O 10 ( C=O-CH2, 2H), 4.354 (N-CH, 1H), 4.022 (-OH, 1H), 3.416-3.553(N-CH2,OH-H, 5H), 2.549-2.909(CH2, CH, 7H), 1.266-2.243(CH2, CH, 49H). 0.893(CH3, 3H).

[0243] Example 3

[0244] This embodiment relates to a method for preparing an estrogen receptor protein-targeted degradation chimeric derivative of general formula (I) based on an estradiol structure, comprising the following steps:

[0245] (1): FWSQ-C5 was obtained by dissolving fulvestrant and dihaloalkanes in a solvent and adding an acid-binding agent under stirring.

[0246] (2) The compound containing the amino group and the E3 ligase ligand end and FWSQ-C5 were dissolved in an organic solvent, and N,N diisopropylethylamine and sodium iodide were added. After the reaction was completed by stirring at 10-35℃, the mixture was extracted and purified to obtain the estrogen receptor protein targeted degradation chimeric derivative of general formula (I) based on the estradiol structure.

[0247] Compound QDE-FWSQ-17

[0248]

[0249] S1: 242.6 mg of fulvestrant (0.4 mmol) and 123.9 mg of 1-bromo-3-iodopropane (0.5 mmol) were dissolved in 3 ml of DMF, and 138 mg of potassium carbonate (1.0 mmol) was added. The reaction was carried out at 25 °C for 12 h, and the reaction progress was monitored by TLC. After the reaction was completed, water / dichloromethane was added for extraction, the dichloromethane phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (DCM:MeOH 40:1-20:1) to obtain 198.4 mg of FWS-C5, with a yield of 68.3%.

[0250] S2: 82.1 mg of 2-(2,6-dioxanediol-3-yl)-5-(3,9-diazaspiro[5.5]undecane-3-yl)isoindole-1,3-dione (0.2 mmol) was dissolved in DMF, and 51.6 mg of DIPEA (0.4 mmol) was added. After stirring for 5 min, 145 mg of 2-(2,6-dioxanediol-3-yl)-5-(3,9-diazaspiro[5.5]undecane-3-yl)isoindole-1,3-dione (0.2 mmol) and 30 mg of NaI (0.2 mmol) were added. The reaction was carried out at 40 °C for 12 h. After the reaction was completed by TLC monitoring, DMF was removed by rotary evaporation, and the mixture was filtered with DCM. The filtrate was concentrated. Purification was performed by silica gel plate preparation (DCM:MeOH 15:1) to obtain 89.9 mg of QDE-FWSQ-17 with a yield of 42.1%.

[0251] The structure of QDE-FWSQ-17 was confirmed by mass spectrometry (HR-ESI-MS) and proton nuclear magnetic resonance (1H-NMR): ESI-MS (positive mode) m / z: [M+H]+1057.5 (calculated for C 57 H 77F5N4O7S, 1056.5433).1H NMR: δ 8.15 (Ph-H, 1H), 8.08 (Ph-H, 1H), 7.00 (Ph-H, 1H), 6.91 (Ph-H, 1H), 6.82 (Ph-H, 1H), 6.67 (Ph-H, 1H), 5.53 (N-CH, 1H), 5.07 (OH, 1H), 3.98 (O-CH2, 2H), 3.65 (OH-CH, 1H), 3.33 (N-CH2, 10H), 3.09 (CH, 1H), 2.84 (CH2, 2H),2.44-2.57 (CH2, 12H), 2.24 (CH2, 2H), 1.76-1.90 (CH2, 8H), 1.64-1.67 (CH2,4H), 1.44-1.55 (CH2, 10H), 1.25-1.31 CH2, (12H), 0.91 (CH, 1H), 0.80 (CH3,3H).

[0252] Compound QDE-FWSQ-19

[0253]

[0254] S1: Take a 10ml single-necked flask, add 96.4mg of tert-butyl 3-(piperazin-1-yl)azacyclobutane-1-carboxylate (0.4mol), dissolve in 2ml of DMSO, add 103.2mg (0.8mmol) of diisopropylethylamine and stir for 5min, then add 110.4mg of 2-(2,6-dioxane-6-hydropyridin-3-yl)-5-fluoroisoindole-1,3-dione (0.4mmol). React at 80℃ for 12h, monitor the reaction progress by TLC, and filter after the reaction is complete. Extracted three times with ethyl acetate and water, the ethyl acetate was separated using a separatory funnel, purified by vacuum distillation and column chromatography (DCM:MeOH 30:1-10:1) to give 167.7 mg of tert-butyl 3-(4-(2-(2,6-dioxopiridin-3-yl)-1,3-dioxoisoindol-5-yl)piperazin-1-yl)azacyclobutane-1-carboxylic acid, i.e., CRBN-19 in 84.4% yield.

[0255] S2: Take a 10ml single-necked flask, add 167.7mg (0.337mmol) of 3-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)piperazin-1-yl)azacyclobutane-1-carboxylic acid tert-butyl ester, dissolve in 5ml of dichloromethane, cool to 0℃ in an ice bath, slowly add 0.5ml of trifluoroacetic acid dropwise to the reaction system, remove the ice bath after the addition is complete, react at room temperature for 3h, quench with sodium bicarbonate solution, extract three times with dichloromethane and water, separate the dichloromethane layer with a separatory funnel, purify by column chromatography under reduced pressure at 30℃ (DCM: MeOH) (10:1) 123.4 mg of compound 5-(4-(azacyclobutan-3-yl)piperazin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione was obtained, with a yield of 92.1%.

[0256] S3: Dissolve 79.4 mg of 5-(4-(azacyclobutan-3-yl)piperazin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (0.2 mmol) in DMF, add 51.6 mg of DIPEA (0.4 mmol), stir for 5 min, then add 145 mg of FWSQ-C5 (0.2 mmol) and 30 mg of NaI (0.2 mmol). React at 40 °C for 12 h. After the reaction is complete as monitored by TLC, remove DMF by rotary evaporation, filter with DCM, and concentrate the filtrate. Silica gel preparative plate purification (DCM:MeOH 15:1) yielded 81.5 mg of 2-(2,6-dioxopiperidin-3-yl)-5-(4-(1-(3-(((7R,8S,9R,13R,14R,17R)-17-hydroxy-13-methyl-7-(9-(4,4,5,5-pentafluoropentyl)sulfinyl)nonyl)-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthrene-3-yl)oxy)propyl)azacyclobutane-3-yl)piperazin-1-yl)isoindole-1,3-dione, i.e., QDE-FWSQ-19, with a yield of 39.1%.

[0257] The structure of QDE-FWSQ-17 was confirmed by mass spectrometry (HR-ESI-MS) and proton nuclear magnetic resonance (1H-NMR): ESI-MS (positive mode) m / z: [M+H]+1044.5 (calculated for C 55 H 74F5N5O7S, 1043.5229).1H NMR: δ 8.15 (Ph-H, 1H), 8.08 (Ph-H,1H), 7.00 (Ph-H,1H), 6.91 (Ph-H,1H), 6.82 (Ph-H,1H), 6.67 (Ph-H,1H), 5.53 (NH, 1H), 5.37 (NH, 1H), 5.07 (OH,1H), 3.98 (N-CH2, 2H), 3.65 (OH-CH, 1H), 3.33 (CH2 / CH, 10H), 3.09 (CH, 1H),2.84 (CH2, 2H), 2.57 (CH2, 4H), 2.44 (CH2, 8H), 2.24 (CH2, 2H), 1.76-1.90 (CH2,8H), 1.64-1.67 (CH2, 4H), 1.44-1.55 (CH2, 10H), 1.25-1.31 (CH2, 12H), 0.91(CH, 1H), 0.80 (CH3, 3H).

[0258] Compound QDE-CEC-4

[0259]

[0260] S1: Take a 25ml single-necked flask, add 54.4mg estradiol (0.2mmol), dissolve in dichloromethane (5ml), then add 68.64mg 1'-(tert-butyloxycarbonyl)-[1,4'-bipiperidine]-4-carboxylic acid (0.22mmol), 40.4mg triethylamine (0.4mmol), and 45.4mg dicyclohexylcarbodiimide (0.22mmol) sequentially. Stir the reaction at room temperature for 24h, monitor the reaction progress by TLC. After the reaction is complete, quench the reaction with saturated ammonium chloride aqueous solution, extract three times with dichloromethane, separate the dichloromethane layer with a separatory funnel, evaporate to dryness under vacuum, and purify by silica gel column chromatography (DCM:MeOH 40:1-10:1) to obtain 46.6mg 4-[4-({[(1S,3aS,3bR,9bS,11aS)-1-hydroxy-11a-methyl-2,3,3a,3b,4,5,9b,10,11,11a-decahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}carbonyl)hexahydropyridin-1-yl]hexahydropyridin-1-carboxylic acid-2-methylpropyl-2-yl ester, i.e., CEC-4-1, yield 41.2%.

[0261] S2: Take a 25ml single-necked bottle and add 46.6mg of compound 4-[4-({[(1S,3aS,3bR,9bS,11aS)-1-hydroxy-11a-methyl-2,3,3a,3b,4,5,9b,10,11,11a-decahydro-1H-cyclopenta[1,2-a]phenanthrene-7-yl]oxy}carbonyl)hexahydropyridin-1-yl]hexahydropyridin-1-carboxylic acid-2-methylpropyl-2- - ester (0.082 mmol) was dissolved in 5 ml of dichloromethane and cooled to 0°C in an ice bath. 0.5 ml of trifluoroacetic acid was slowly added dropwise to the reaction system. After the addition was complete, the ice bath was removed, and the reaction was carried out at room temperature for 3 hours. The reaction progress was monitored by TLC. After the reaction was complete, sodium bicarbonate solution was added to quench the reaction. The mixture was extracted three times with dichloromethane and water. The dichloromethane layer was separated using a separatory funnel, and purified by column chromatography under reduced pressure at 30°C (DCM: MeOH). (10:1) 34.6 mg of 1-(hexahydropyridin-4-yl)hexahydropyridin-4-carboxylic acid-(1S,3aS,3bR,9bS,11aS)-1-hydroxy-11a-methyl-2,3,3a,3b,4,5,9b,10,11,11a-decahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester, i.e., CEC-4-2, was obtained with a yield of 90.3%.

[0262] S3: Take a 10ml single-necked flask, add 24.6mg of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)acetic acid (0.074mmol), add 2ml of DCM suspension, add 28.1mg (0.074mmol) of HATU and 19mg (0.148mmol) of diisopropylethylamine, stir and activate for 30min, add 34.6mg of CEC-4-2 (0.074mmol), react at room temperature for 3h, monitor the reaction progress by TLC, after the reaction is complete, extract with ethyl acetate and water, extract three times, separate ethyl acetate with a separatory funnel, purify by vacuum distillation and column chromatography (DCM: MeOH). (30:1-10:1) yielded 19.2 mg of compound 1-[1-(2-{[2-(2,6-dioxylidenehexahydropyridin-3-yl)-1,3-dioxylidene-2,3-dihydro-1H-isoindol-4-yl]oxy}acetyl)hexahydropyridin-4-yl]hexahydropyridin-4-carboxylic acid-(1S,3aS,3bR,9bS,11aS)-1-hydroxy-11a-methyl-2,3,3a,3b,4,5,9b,10,11,11a-decahydro-1H-cyclopenta[1,2-i]phenanthrene-7-yl ester, i.e., QDE-CEC-4, yield 33.2%.

[0263] The structure of QDE-CEC-4 was confirmed by mass spectrometry (HR-ESI-MS) and proton nuclear magnetic resonance (1H-NMR): ESI-MS (positive mode) m / z: [M+H]+ = 781.3 (calculated for C 44 H 52 N4O9, 780.3734). 1HNMR: δ 7.86 (Ph-H, 1H), 7.24 (Ph-H, 1H), 7.00 (Ph-H, 1H), 6.91 (Ph-H, 1H), 6.89 (Ph-H, 1H), 6.82 (Ph-H, 1H), 5.53 (N-CH, H), 5.37 (N-CH, 1H), 5.07 (N-CH, 1H), 4.36 (CO-CH2, 2H), 3.65-3.76 (O-CH2, 2H), 3.33 (N-CH2, 8H), 3.09 (OH,1H), 3.00 (CH2, 4H), 2.83 (CH2, 2H), 2.34 (CH , 1H), 2.24 (CH2, 2H), 1.97 (CH2,2H), 1.77-1.78 (CH2, 4H), 1.65 (CH2, 4H), 1.47-1.56 (CH2 / CH, 8H), 0.80 (CH3,3H).

[0264] Test Example 1: Assay of Estrogen Receptor Protein Hydrolysis Activity

[0265] The estrogen receptor protein hydrolysis activity of the estrogen receptor protein-targeting degradation chimeric derivatives based on the estradiol structure prepared in the above examples was tested, and QDE-003-C in patent CN118027003A was used as a control. The experimental procedure is as follows:

[0266] 1. Take MCF-7 cells in logarithmic growth phase, seed them into 6-well plates at a rate of 1.5 × 10⁶ cells / well, and incubate overnight;

[0267] 2. Add positive control and test group (DMSO ratio of 0.1%) to a final concentration of 1 μM and incubate in an incubator for 24 h;

[0268] 3. Collect cells from each group and wash once with PBS; add cell lysis buffer containing 1×Protease Inhibitor Cocktail and 1×Phosphatase Inhibitor Cocktail II, lyse thoroughly on ice, centrifuge at 14000 rpm for 10 min at 4°C and collect the supernatant.

[0269] 4. After determining the protein concentration using the BCA kit, the Estrogen Receptor alpha protein level was detected by Western blot.

[0270] The test results are shown in Table 1. A schematic diagram of the Western blot experiment showing the inhibition of ER protein levels in MCF-7 cells by the estrogen receptor protein-targeted degradation chimeric derivative based on the estradiol structure is shown in Table 1. Figure 1-3 The WB assay for ER protein degradation yielded a quantitative protein concentration graph. Figure 4 , Figure 1-3 In the figure "17: Fulvestrant 0.1μM", the control group for fulvestrant is shown. It can be observed that at the same administered concentration (0.1μM), the IntDen value to internal control (GAPDH) ratio of estrogen receptor protein (ER) of the four compounds QDE-FWSQ-3, QDE-FWSQ-4, QDE-FWSQ-12, and QDE-FWSQ-21 is lower than that of the positive control drug Fulvestrant. This demonstrates that their ability to degrade estrogen receptors in MCF-7 breast cancer cells is superior to that of the positive control drug Fulvestrant.

[0271] Table 1 Results of estrogen receptor protein hydrolysis activity assay

[0272]

[0273] The above description is merely an embodiment of this application, and the scope of protection of this application is not limited to these specific embodiments, but is determined by the claims of this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the technical concept and principles of this application should be included within the scope of protection of this application.

Claims

1. A chimeric derivative of estrogen receptor protein targeting degradation based on an estradiol structure, characterized in that, It has the structure of general formula (I): ABC (I) Where A is an E3 ligase ligand, selected from one of general formulas (II-1), (II-2), or (II-3): Equation (Ⅱ-1); Equation (Ⅱ-2); Equation (Ⅱ-3); * indicates a connection site, n=0 or 1, m=0 or 1, in formula (II-1), general formula (II-2) and general formula (II-3) R1, R2 and R3 are each independently one of: H, OH, F, Cl, Br, NH2, CH3 or OCH3, and R4 is hydrogen or methyl. B is selected from one of the following formulas: ； C has the structure of general formula (Ⅲ): Formula (III).

2. The estrogen receptor protein-targeted degradation chimeric derivative based on the estradiol structure according to claim 1, characterized in that, The estrogen receptor protein targeted degradation chimeric derivative based on the estradiol structure includes its cis-trans isomer and racemic isomer.

3. A chimeric derivative of estrogen receptor protein targeting degradation based on an estradiol structure, characterized in that, It has one of the following structural formulas: ； ； ； ； ； ； ； ； ; ； 。 4. The method for preparing the estrogen receptor protein-targeted degradation chimeric derivative based on the estradiol structure according to any one of claims 1-3, characterized in that, Includes the following steps: (1) Place fulvestrant in a solvent, add an acid-binding agent and lower the temperature to 1-5℃, then add a monomer containing halogen atoms and acyl halide groups to react and obtain compound 2; (2) Compound 2 is reacted sequentially with a monomer containing amino and tert-butoxycarbonyl groups and an E3 ligase ligand A containing a flexible linker to obtain an estrogen receptor protein-targeting degradation chimeric derivative of general formula (I) based on an estradiol structure, wherein the E3 ligase ligand A containing a flexible linker has a carboxyl group or a ketone carbonyl group at its end; or Compound 2 was reacted with E3 ligase ligand B with a flexible linker to obtain an estrogen receptor protein targeted degradation chimeric derivative of general formula (I) based on an estradiol structure, wherein the E3 ligase ligand B with the flexible linker has an amino group at its end.

5. The preparation method according to claim 4, characterized in that, Step (2) involves reacting compound 2 with a monomer containing amino and tert-butoxycarbonyl groups and E3 ligase ligand A containing a flexible linker in sequence to obtain the estrogen receptor protein targeted degradation chimeric derivative of general formula (I) based on an estradiol structure. Specifically: (2-1): Compound 2 is condensed with a monomer containing amino and tert-butoxycarbonyl groups to obtain intermediate A; (2-2): The protecting group of intermediate A is removed under acidic conditions to obtain intermediate B containing an amino group; (2-3): Intermediate B undergoes a condensation reaction with E3 ligase ligand A with a flexible linker to obtain a chimeric derivative of estrogen receptor protein targeting degradation based on the estradiol structure of general formula (I).

6. The method for preparing the estrogen receptor protein-targeted degradation chimeric derivative based on the estradiol structure according to any one of claims 1-3, characterized in that, Includes the following steps: S1: Fluvestrant is stirred with a monomer containing carboxyl and tert-butyloxycarbonyl groups in the presence of a condensing agent and a base to obtain an intermediate containing tert-butyloxycarbonyl protection. S2: The intermediate containing tert-butyloxycarbonyl protected group obtained in S1 is placed in an acidic environment to remove the protecting group and obtain an intermediate containing an amino group. S3: The amino-containing intermediate obtained in S2 is reacted with E3 ligase ligand A containing a carboxyl group, and the mixture is extracted and purified to obtain a chimeric derivative of estrogen receptor protein targeting degradation based on the estradiol structure of general formula (I). The E3 ligase ligand A with a flexible linker has a carboxyl group or a ketone carbonyl group at its end.

7. The method for preparing the estrogen receptor protein-targeted degradation chimeric derivative based on the estradiol structure according to any one of claims 1-3, characterized in that, Includes the following steps: D1: Dissolve fulvestrant and dihaloalkane in a solvent, add an acid-binding agent and stir to obtain FWSQ-C5; D2: The compound containing the amino group and the E3 ligase ligand end and FWSQ-C5 were dissolved in an organic solvent, N,N diisopropylethylamine and sodium iodide were added, and after stirring and reaction was completed, the mixture was extracted and purified to obtain the estrogen receptor protein targeted degradation chimeric derivative of general formula (I) based on the estradiol structure.

8. A pharmaceutical composition, characterized in that, The composition contains: a therapeutically effective amount of any one of the estrogenic triene-based estrogen receptor protein targeted degradation chimeric derivatives or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier thereof.

9. The use of any one of the estrogen receptor protein targeted degradation chimeric derivatives based on the estradiol structure of any one of claims 1-3 or the pharmaceutical composition of claim 8 in the preparation of a medicament for the prevention and treatment of breast cancer.

10. The use of any one of the estrogen receptor protein targeted degradation chimeric derivatives based on the estradiol structure of any one of claims 1-3 or the pharmaceutical composition of claim 8 in the preparation of a medicament for treating diseases related to estrogen receptor protein targeted degradation chimeric.

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