Novel AR degradation agent as well as preparation method and application thereof

Abstract

The invention discloses a novel AR degradation agent as well as a preparation method and application thereof. The AR degradation agent is PROTAC with GROs (such as AS1411) as an MDM2 recruitment element, target protein AR and E3 ubiquitin ligase MDM2 can be drawn close in vivo, the AR is labeled with ubiquitin, and then the AR is degraded through a ubiquitin-proteasome pathway. Experiments show that the AR degradation agent can effectively degrade AR and generate corresponding curative effects (such as inhibition of tumor cell proliferation and promotion of tumor cell apoptosis), and has very good application prospects and research values.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a novel AR degradation agent and a preparation method and application thereof. Background Art

[0002] Androgen receptor (AR) belongs to the steroid receptor superfamily and is a ligand-dependent nuclear transcription factor. AR consists of four functional domains, including the N-terminal transcription activation domain (NTD), DNA binding domain (DBD), hinge region with nuclear localization signal and C-terminal ligand binding domain (LBD). AR plays an important role in promoting and maintaining reproductive, musculoskeletal, immune, cardiovascular, hematopoietic and nervous system functions. In the absence of ligand, AR mainly exists in the cytoplasm in the form of binding to molecular chaperones (such as heat shock proteins HSP70, HSP90 and other chaperone proteins), thereby inhibiting the transcription of downstream genes; when the LBD of AR binds to androgen, AR dimerizes and transfers to the nucleus under the action of coactivators, binds to androgen response elements (ARE), and regulates the transcription of downstream target genes. AR is widely present in all tissues of the body and plays a vital role in the growth and development of the human body. However, abnormal expression of AR is closely related to the occurrence and development of various tumors (such as prostate cancer, breast cancer, bladder cancer, liver cancer, kidney cancer, lung cancer, endometrial cancer, ovarian cancer and pancreatic cancer, etc.) and hormone-related diseases (such as androgenic alopecia, etc.).

[0003] So far, a variety of AR-targeted drugs have been approved for the treatment of AR-related diseases. For example, the first-generation AR inhibitors (bicalutamide, flutamide, and nilutamide) bind to the LBD of AR, resulting in inhibition of androgen binding to the LBD, thereby alleviating the symptoms of prostate cancer patients. Recently, the second-generation AR inhibitors have been developed and used clinically. For example, enzalutamide can inhibit androgen receptor nuclear translocation, transcriptional binding, and recruitment of co-activators; apalutamide inhibits AR nuclear translocation, inhibits DNA binding, and hinders AR-mediated transcription, thereby effectively delaying the progression of prostate cancer. However, AR inhibitors will inevitably cause drug resistance, among which an important mechanism of drug resistance is the mutation of the AR gene, including AR amplification, AR mutation, and AR splice variants. Moreover, the inhibition of AR protein requires that the drug be maintained at a high concentration for a long time. High-dose administration will lead to AR protein enrichment due to negative feedback, thereby greatly weakening its inhibitory effect. Therefore, it is necessary to further develop new drugs that are more effective against AR protein.

[0004] Targeted protein degradation is a new breakthrough drug development strategy that uses the inherent protein degradation pathway in cells to directly degrade pathogenic target proteins. This new drug form includes many types, such as PROTAC, molecular glue, LYTAC, ATAC, AbTAC, ATTEC, AUTAC, AUTOTAC, etc. PROTAC (PROteolysis Targeting Chimera) is a bifunctional molecule composed of three parts: target protein ligand, linker and E3 ubiquitin ligase recruitment element. After PROTAC enters the cell, the target protein ligand in its structure specifically binds to the target protein, and the E3 ligase recruitment element at the other end binds to the E3 ligase to form a target protein-PROTAC-E3 ternary complex. The E3 ubiquitin ligase mediates the ubiquitin conjugation enzyme E2 to ubiquitinate the target protein, and the target protein marked by polyubiquitination will be transported to the proteasome for degradation, thereby reducing the level of the target protein. In the above process, the target protein ligand does not need to occupy the binding site for a long time. Therefore, PROTAC can play a role in multiple cycles in the cell. Based on the special mechanism of action of PROTAC, PROTAC drugs have important advantages in drug development for overcoming drug resistance and undruggable targets. Summary of the invention

[0005] In order to overcome the deficiencies of the prior art, the present invention provides a novel AR degrader and its preparation method and application. The AR degrader is a PROTAC with GROs (such as AS1411) as the MDM2 recruitment element, which can bring the target protein AR and the E3 ubiquitin ligase MDM2 closer in vivo, so that AR is labeled with ubiquitin and then degraded through the ubiquitin-proteasome pathway. Experiments have shown that the AR degrader can effectively degrade AR and produce corresponding therapeutic effects, such as inhibiting the proliferation of tumor (such as prostate cancer) cells and promoting tumor cell apoptosis.

[0006] In a first aspect of the present invention, an AR degradation agent and a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate or deuterated compound thereof are provided, wherein the AR degradation agent has the following structure: Wherein, GRO is a guanine-rich oligonucleotide (G-rich oligonucleotides) residue that can specifically bind to nucleolin (NCL), L is a connecting structure part, ARB is an androgen receptor recognition / binding part, and p is any suitable integer within 1-100.

[0007] Specifically, the ARB can be any suitable structural portion of an androgen receptor ligand, such as the ARB portion described in CN111825657A (specifically as shown in claims 6-7 thereof).

[0008] In particular, the AR degrading agent has the following structure:

[0009]

[0010] Among them, GRO is a guanine-rich oligonucleotide (G-rich oligonucleotides) residue that can specifically bind to nucleolin (NCL);

[0011] L is a linker;

[0012] p is an integer from 1 to 100 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 15, 20, 25, 30, 40, 50, 60, 80, 100);

[0013] Y 1 , Y 2 , Y 3 are independently selected from: a single bond, -O-, -S-, -N(C 0 -C 6 alkyl), -C(O)-, -C(S)-, -C(O)-(C 0 -C 6 Alkylene)-, -C(S)-(C 0-C 6 Alkylene)-, -C(O)-N(C 0 -C 6 Alkyl)-, -C(S)-N(C 0 -C 6 Alkyl)-, SO, SO 2 ;

[0014] Ring A is an aromatic ring or an aromatic heterocyclic ring;

[0015] Ring B is an aliphatic ring or a heterocyclic ring;

[0016] Ring C is an aromatic ring or an aromatic heterocyclic ring;

[0017] R 1 is one or more independent substituents on the A ring selected from: H, C 1 -C 10 Alkyl, -(C 0 -C 6 Alkylene)-(C 3 -C 10 Cycloalkyl), -(C 0 -C 6 Alkylene)-(C 6 -C 10 Aryl), -(C 0 -C 6 alkylene)-(4-10 membered heterocyclic group), halogen, cyano, nitro, azido, C 1 -C 10 Haloalkyl, C 1 -C 10 Haloalkoxy, -N(C 0-10 Alkyl)(C 0-10 Alkyl), -N(C 0-10 alkyl)CO(C 0-10 Alkyl), -N(C 0-10 alkyl)CON(C 0-10 Alkyl), -N(C 0-10 alkyl)SO 2 (C 0-10 Alkyl), -O(C 0-10 Alkyl), -S(C 0-10 Alkyl), -SO(C 0-10 Alkyl), -SO 2 (C 0-10 Alkyl), -SO 2 N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C 0-10 Alkyl)(C0-10 Alkyl), -CO(C 0-10 alkyl);

[0018] R 2 is one or more independent substituents on the B ring selected from: H, C 1 -C 10 Alkyl, -(C 0 -C 6 Alkylene)-(C 3 -C 10 Cycloalkyl), -(C 0 -C 6 Alkylene)-(C 6 -C 10 Aryl), -(C 0 -C 6 alkylene)-(4-10 membered heterocyclic group), halogen, cyano, nitro, azido, C 1 -C 10 Haloalkyl, C 1 -C 10 Haloalkoxy, -N(C 0-10 Alkyl)(C 0-10 Alkyl), -N(C 0-10 alkyl)CO(C 0-10 Alkyl), -N(C 0-10 alkyl)CON(C 0-10 Alkyl), -N(C 0-10 alkyl)SO 2 (C 0-10 Alkyl), -O(C 0-10 Alkyl), -S(C 0-10 Alkyl), -SO(C 0-10 Alkyl), -SO 2 (C 0-10 Alkyl), -SO 2 N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C 0-10 Alkyl)(C 0-10 Alkyl), -CO(C 0-10 alkyl);

[0019] R 3 is one or more independent substituents on the C ring selected from: H, C 1 -C 10 Alkyl, -(C 0 -C 6 Alkylene)-(C 3 -C 10Cycloalkyl), -(C 0 -C 6 Alkylene)-(C 6 -C 10 Aryl), -(C 0 -C 6 alkylene)-(4-10 membered heterocyclic group), halogen, cyano, nitro, azido, C 1 -C 10 Haloalkyl, C 1 -C 10 Haloalkoxy, -N(C 0-10 Alkyl)(C 0-10 Alkyl), -N(C 0-10 alkyl)CO(C 0-10 Alkyl), -N(C 0-10 alkyl)CON(C 0-10 Alkyl), -N(C 0-10 alkyl)SO 2 (C 0-10 Alkyl), -O(C 0-10 Alkyl), -S(C 0-10 Alkyl), -SO(C 0-10 Alkyl), -SO 2 (C 0-10 Alkyl), -SO 2 N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C 0-10 Alkyl)(C 0-10 Alkyl), -CO(C 0-10 alkyl).

[0020] In a preferred embodiment of the present invention, p is 1, that is, the general formula I is

[0021] Specifically, the GROs are guanine-rich oligonucleotides containing one or more GGT motifs and having G4 structural features.

[0022] Preferably, the GROs have a stable G4 structural feature, wherein the G4 structural characteristic signal can be detected by the method described in the prior art (see, for example, Yu Yuan, Hu Fang, Xia Yuan, et al. G-quadruplex detection methods and biological research progress [J]. Chemistry of Life. 2021, 14(10): 2146-2155.; Gao Juan, Yuan Gu, Xu Ming. Detection, function and regulation of G-quadruplex structure [J]. Progress in Physiological Sciences, 2014, 45(5): 364-371.), for example, using probes (such as specific antibodies, specific fluorescent ligands, radioactive markers, etc.) for detection, or using circular dichroism (CD), nuclear magnetic resonance (NMR), ultraviolet spectroscopy, molecular fluorescence spectroscopy, single-molecule fluorescence resonance energy transfer (FRET), etc. for detection.

[0023] In some embodiments of the present invention, fluorescent probes (such as N-methylporphyrin dipropionic acid IX (NMM), O-phenanthroline derivative) are used for detection, and the GROs have a stable G4 structural characteristic signal (see, for example, Zhang Suge, Sun Hongxia, Tang Yalin. Research progress of DNA G-quadruplex recognition probes [J]. Chemical Bulletin, 2016, 79(5): 387-394.).

[0024] In one embodiment of the present invention, when N-methylporphyrin dipropionic acid IX (NMM) is used as a fluorescent probe for detection, in the presence or absence of K + In the presence of AS1411, a stable G4 structural feature can be formed. Specifically, the detection method may include the following steps: adding the GRO to be tested with or without K + The solution was heated at 90-99° C. for 5-10 minutes, incubated on ice, and then NMM was added. The mixture was incubated at room temperature in the dark, and the fluorescence intensity was detected (such as described in Example 1 of the present invention).

[0025] Specifically, the GROs include chemical modifications, nucleic acid unit replacement or functional group connection on GROs; these modifications or functional groups can be used to improve the stability of PROTAC, provide detection signals, or form compositions with other substances. Wherein, the chemical modification is that at least one base is modified, and the chemical modification includes at least one of phosphorylation, methylation, amination, sulfhydrylation, isotopization, thiophosphate backbone modification, methoxy modification, and fluorination modification; the nucleic acid unit is replaced with at least one nucleic acid unit replaced with LNA, UNA or GNA; the functional group includes at least one of a fluorescent group, a radioactive group, a therapeutic drug, biotin, digoxin, a nanoluminescent material, a nucleic acid substance or an enzyme marker.

[0026] Preferably, the GROs have 4-100 (e.g., 4, 5, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 40, 50, 60, 80, 100) nucleotides.

[0027] Preferably, the GROs is one of the DNA aptamer AS1411 and AS1411 derivatives / analogues.

[0028] Among them, the AS1411 derivatives / analogues are GROs with a homology of more than 60% (e.g., 65%, 70%, 75%, 80%, 85%, 90%, 95% or more) with AS1411, and contain one or more GGT motifs, have G4 structural features, and can specifically bind to NCL.

[0029] Specifically, the AS1411 derivative / analogue is one of GRO29A, GRO15A, AT11, AT11-L0, AT21, AT27, GRO29-2, GRO29-3, GRO29-5, GRO29-13, GRO14C, GRO25A, GRO28A, GRO28B, GRO29-6, GRO32A, GRO32B, GRO56A, GRO A, GRO B, GRO C, GRO D, GRO E, GRO F, GRO G, GRO H, GRO I, GRO J, GROK, GRO L, and GRO M, and their nucleotide sequences are shown in SEQ ID NOs: 8-38, respectively.

[0030] Specifically, the sequence of AS1411 is shown in SEQ ID NO: 1: 5′-GGTGGTGGTGGTTGTGGTGGTGGTGG-3′.

[0031] In some preferred embodiments of the present invention, the GROs is AS1411.

[0032] In other embodiments of the present invention, the GROs are selected from: GRO29A, GRO15A, AT11.

[0033] Specifically, L can be any suitable linking moiety that connects the GRO residue to the AR recognition / binding moiety.

[0034] Specifically, L can be linked to any base, sugar or phosphate backbone at the 3' end, 5' end or in the middle of GRO.

[0035] In some embodiments of the invention, L is attached to the 3' end of GRO.

[0036] In some embodiments of the invention, L is attached to the 5' end of GRO.

[0037] In some embodiments of the present invention, the GRO moiety in Formula I has the following structure:

[0038]

[0039] Specifically, L has the following structure: in,

[0040] L 1 is a divalent group connected to GRO, which can be selected from: a single bond, -O-(C 0 -C 6 Alkylene)-, -S-(C 0 -C 6 Alkylene)-, -N(R L1 )-(C 0 -C 6 Alkylene)-, -N(R L2 )C(O)-(C 0 -C 6 Alkylene)-, -OP(O)(OR L1 )O-(C 0 -C 6 Alkylene)-, -C(O)-(C 0 -C 6 Alkylene)-, -C(S)-(C 0 -C 6 Alkylene)-, -CON(R L1 )-(C 0 -C 6 Alkylene)-;

[0041] L 3 is a divalent group connected to the A ring, which can be selected from: a single bond, -O-(C 0 -C 6 Alkylene)-, -S-(C 0 -C 6 Alkylene)-, -C(O)-(C 0 -C 6 Alkylene)-, -C(S)-(C 0 -C 6 Alkylene)-, -N(R L3 )-(C 0 -C 6 Alkylene)-, -CON(R L3 )-(C 0 -C 6 Alkylene)-, -N(R L3 )CO-(C 0-C 6 Alkylene)-, -SO 2 -(C 0 -C 6 Alkylene)-, -SO-(C 0 -C 6 Alkylene)-, 4-10 membered heterocyclylene;

[0042] L 2 is a single bond or a divalent saturated or unsaturated straight or branched C1-C50 hydrocarbon chain (e.g., a C1-C20 alkyl chain), wherein 0-6 methylene units in the hydrocarbon chain (e.g., an alkyl chain) are independently substituted by: -CY-, -O-, -S-, -SS-, -C(O)-, -C(S)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(O)N(R L2 )-、-N(R L2 )C(O)-、-N(R L2 )C(O)O-、-N(R L2 )C(O)N(R L2 )-、-N(R L2 )-、-S(O) 2 -、-S(O) 2 N(R L2 )-、-N(R L2 )S(O) 2 -、-S(O)-、-S(O)N(R L2 )-、-N(R L2 )S(O)-、-P(O)(OR L2 )O-、-P(O)-、-P(O)N(R L2 )-、-P(O)(N(R L2 ) 2 )-、-OP(O)(OR L2 ) 2 N(R L2 )-、-P(O)(OR L2 ) 2 N(R L2 )-、-N(R L2 )P(O)(OR L2 )O-、-N(R L2 )P(O)-、-Si(R L2 ) 2 -、-C(=N-CN)-、

[0043] Amino acid residues, nucleotide residues, oligonucleotide residues, oligopeptide residues, wherein m2 is selected from an integer selected from 1-10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10), each -CY- is independently an optionally substituted divalent ring selected from the following: arylene, cycloalkylene, heterocyclylene; H in the hydrocarbon chain may be optionally substituted by one or more groups selected from the following: halogen, cyano, nitro, azido, -OR L0 、-C(O)R L0 、-C(S)R L0 、-C(O)OR L0 、-C(S)SR L0 、-OC(O)R L0 、-OC(S)R L0 、-OC(S)SR L0 、-C(O)N(R L0 ) 2 、-OC(O)N(R L0 ) 2 、-N(R L0 )C(O)OR L0 、-N(R L0 )SO 2 R L0 、-SO 2 N(R L0 ) 2 、-OSO 2 N(R L0 ) 2 、-N(R L0 )C(O)R L0 、-N(R L0 ) 2 、-SR L0 、-SOR L0 、-SO 2 R L0 、-OSO 2 R L0 , C 1 -C 10 Alkyl, C 2 -C 10 Alkenyl, C 2 -C 10 Alkynyl, C 1 -C 10 Haloalkyl, C 1 -C 10 Haloalkoxy, -(C 0 -C 6 Alkylene)-(C 3 -C 10 Cycloalkyl), -(C 0 -C 6Alkylene)-(C 6 -C 10 Aryl), -(C 0 -C 6 Alkylene)-(4-10 membered heterocyclyl);

[0044] R L0 , R L1 , R L2 and R L3 Independently selected from: H, C 1 -C 10 Alkyl, C 2 -C 10 Alkenyl, C 2 -C 10 Alkynyl, -(C 0 -C 6 Alkylene)-(C 3 -C 10 Cycloalkyl), -(C 0 -C 6 Alkylene)-(C 6 -C 10 Aryl), -(C 0 -C 6 alkylene)-(4-10 membered heterocyclic group), wherein the C 1 -C 10 Alkyl, C 2 -C 10 Alkenyl, C 2 -C 10 Alkynyl, C 0 -C 6 Alkylene, C 3 -C 10 Cycloalkyl, C 6 -C 10 The H in the aryl group and the 4-10 membered heterocyclic group may be optionally substituted by one or more groups selected from the following: halogen, cyano, nitro, azido, hydroxyl, amino, thiol, carboxyl, C 1 -C 10 Alkyl, C 2 -C 10 Alkenyl, C 2 -C 10 Alkynyl, C 1 -C 10 Haloalkyl, C 1 -C 10 Haloalkoxy, -(C 0 -C 6 Alkylene)-(C 3 -C 10 Cycloalkyl), -(C 0 -C 6 Alkylene)-(C6 -C 10 Aryl), -(C 0 -C 6 alkylene)-(4-10 membered heterocyclic group).

[0045] Specifically, each -CY- is independently selected from the following optionally substituted bivalent rings: phenylene, bicyclic arylene, tricyclic arylene, monocyclic cycloalkylene, bicyclic cycloalkylene, tricyclic cycloalkylene, monocyclic heteroarylene, bicyclic heteroarylene, tricyclic heteroarylene, monocyclic heterocycloalkylene, bicyclic heterocycloalkylene, tricyclic heterocycloalkylene.

[0046] In some embodiments of the invention, each -CY- is independently selected from the following:

[0047]

[0048]

[0049] Among them, R L4 , R L5 Independently selected from: H, OH, halogen, C 1-8 Alkyl, O(C 1-8 Alkyl), S(C 1-8 Alkyl), NH(C 1-8 Alkyl), N(C 1-8 alkyl) 2 , C 3-11 Cycloalkyl, C 3-11 Heterocyclic hydrocarbon, O(C 1-8 Cycloalkyl), S(C 1-8 Cyclic hydrocarbon), NH(C 1-8 Cycloalkyl), N(C 1-8 Cycloalkyl) 1-8 alkyl), OH, NH 2 , SH, SO 2 (C 1-8 alkyl), P(=O)(OC 1-8 Alkyl)(C 1-8 alkyl), P(=O)(OC 1-8 alkyl) 2 , C 1-8 Alkynyl, CH=CH(C 1-8 alkyl), C(C 1-8 alkyl)=CH(C 1-8 alkyl), C(C 1-8 alkyl)=C(C 1-8 alkyl) 2 、Si(OH)3 、Si(C 1-8 alkyl) 3 、Si(OH)(C 1-8 alkyl) 2 , C(=O)(C 1-8 Alkyl), CO 2 H, CN, CF 3 , CHF 2 , CH 2 F. NO 2 , SF 5 、SO 2 NH(C 1-8 alkyl), SO 2 N(C 1-8 alkyl) 2 、S(=O)N(C 1-8 alkyl) 2 、C(=O)NH(C 1-8 alkyl), C(=O)N(C 1-8 alkyl) 2 、N(C 1-8 alkyl)C(=O)NH(C 1-8 Alkyl), N(C 1-8 alkyl)C(=O)N(C 1-8 alkyl) 2 、NHC(=O)NH(C 1-8 alkyl), NHC(=O)N(C 1-8 alkyl) 2 、NHC(=O)NH 2 、N(C 1-8 alkyl)SO 2 NH(C 1-8 Alkyl), N(C 1-8 alkyl)SO 2 N(C 1-8 alkyl) 2 、NHSO 2 NH(C 1-8 Alkyl), NHSO 2 N(C 1-8 alkyl) 2 or NHSO 2 NH 2 or, R L4 , R L5 Together with the atoms to which it is attached, it forms a cycloalkylene or heterocyclylene.

[0050] More specifically, R L4 , R L5 Independently selected from: -CH 3 , -OH,

[0051]

[0052]

[0053] or, R L4 , R L5 Together with the atoms to which they are attached, they form a three-membered to six-membered cycloalkylene group (e.g. ) or a four-membered to six-membered heterocycloalkylene group (such as ).

[0054] In some embodiments of the present invention, R L1 For H.

[0055] In some embodiments of the present invention, R L3 For H.

[0056] In some embodiments of the present invention, R L4 For H.

[0057] In some embodiments of the present invention, R L4 For OH.

[0058] In some embodiments of the present invention, R L5 For H.

[0059] In some embodiments of the present invention, R L5 For OH.

[0060] In one embodiment of the present invention, L 2 The following scheme (1) is adopted: L 2 is a C1-C20 straight chain alkylene group, wherein 0-6 methylene units in the alkylene group are independently substituted by the following groups: -O-, -S-, -SS-, -C(O)-, -C(O)O-, -OC(O)-, -N(R L2 )-、-C(O)N(R L2 )-、-N(R L2 )C(O)-、

[0061]

[0062] Among them, each R L2 Independently selected from: H, C 1 -C 6 Alkyl, each R L4 Independently selected from: H, OH, C 1 -C 6 Alkoxy.

[0063] More specifically, L 2 Can be selected from: C1-C20 straight chain alkylene, -(CH 2 CH 2 O) m2 -CH 2 -、-(CH 2 CH 2 O) m2 -CH 2 CH 2 -、-CH 2 -(CH 2 CH 2 O) m2 -CH 2 -、-CH 2 CH 2 -(CH 2 CH 2 O) m2 -CH 2 -、-CH 2 CH 2 -(CH 2 CH 2 O) m2 -CH 2 CH 2 -、-(C 1 -C 10 Alkylene)-O-(C 1 -C 10 Alkylene)-, -(C 1 -C 10 Alkylene)-NH-(C 1 -C 10 Alkylene)-, -(C 1 -C 10 Alkylene)-C(O)NH-(C 1 -C 10 Alkylene)-, -(C 1 -C 10 Alkylene)-NHC(O)-(C 1 -C 10 Alkylene)-, -(C 1 -C 6 Alkylene)-O-(C 1 -C 6 Alkylene)-C(O)NH-(C 1 -C 6 Alkylene)-, -(C 1 -C 6 Alkylene)-O-(C 1 -C 6 Alkylene)-NHC(O)-(C1 -C 6 Alkylene)-,

[0064]

[0065] Wherein, m2 is selected from an integer between 1 and 10, g is 0 or 1, h is selected from an integer between 0 and 10 (for example, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10), i is selected from an integer between 0 and 10 (for example, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10), and G is any suitable trivalent group.

[0066] In some specific embodiments of the present invention, L 2 Selected from:

[0067] In some embodiments of the present invention, L 2 Selected from C1-C20 straight chain alkylene, -(C 0 -C 6 Alkylene)-(CH 2 CH 2 O) m2 -(C 1 -C 6 Alkylene)-, -(C 1 -C 10 Alkylene)-O-(C 1 -C 10 Alkylene)-, -(C 1 -C 10 Alkylene)-NH-(C 1 -C 10 Alkylene)-, -(C 1 -C 10 Alkylene)-C(O)NH-(C 1 -C 10 Alkylene)-, -(C 1 -C 10 Alkylene)-NHC(O)-(C 1 -C 10 Alkylene)-, -(C 1 -C 6 Alkylene)-O-(C 1 -C 6 Alkylene)-C(O)NH-(C 1 -C 6 Alkylene)-, -(C 1 -C 6 Alkylene)-O-(C 1 -C 6 Alkylene)-NHC(O)-(C1 -C 6 alkylene)-, wherein one methylene unit in the alkylene is independently connected to a group that can be connected to a solid support (e.g. G is any suitable trivalent group).

[0068] In some embodiments of the present invention, for Wherein, the J ring is a 4-10 membered saturated heterocyclic ring.

[0069] Specifically, the J ring is a 4-6 membered saturated heterocyclic ring, which optionally contains other heteroatoms, for example More specifically, It can be, for example,

[0070] In some embodiments of the invention, G is a sugar residue, such as Can be

[0071] In some embodiments of the present invention, L 2 for in particular (x end connected to L 3 , y end connected to L 1 ),For example

[0072] In another embodiment of the present invention, L 2 The following scheme (2) is adopted: L 2 is a C1-C20 straight chain alkylene group, wherein 1-3 methylene units are independently substituted by the following groups: -CY-, Optionally, L 2 The group also includes a group selected from the following: -O-, -C(O)-, -N(R L2 )-、-C(O)N(R L2 )-、-N(R L2 )C(O)-、

[0073]

[0074] Among them, each R L2 Independently selected from: H, C 1 -C 6 Alkyl, each R L4 Independently selected from: H, OH, C 1 -C 6 Alkoxy.

[0075] In some embodiments of the invention, -CY- is selected from:

[0076] In some specific embodiments of the present invention, L 2 Selected from:

[0077]

[0078] In another embodiment of the present invention, L 2 The following scheme (3) is adopted: L 2 L is an oligonucleotide residue, such as a DNA oligonucleotide residue, an RNA oligonucleotide residue, or a DNA / RNA hybrid oligonucleotide residue, which may be single-stranded or double-stranded. 1 Can be a single bond or O, i.e. GRO Through the phosphate bond (i.e. ) and L 2 connect.

[0079] Specifically, the oligonucleotide consists of A and / or T.

[0080] In some embodiments of the present invention, L 2 It is a single-stranded oligonucleotide residue comprising 3-30 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30) A or T, for example, a single-stranded oligonucleotide residue consisting of 6 A.

[0081] In some embodiments of the present invention, L 2 It is a double-stranded oligonucleotide residue comprising 3-30 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30) AT pairs, for example, a double-stranded oligonucleotide residue consisting of 6 AT pairs.

[0082] In some embodiments of the present invention, L 2 It is a single-stranded DNA oligonucleotide residue consisting of 6 A's.

[0083] In some embodiments of the present invention, L 2 It is a double-stranded DNA oligonucleotide composed of 6 AT pairs.

[0084] In some embodiments of the present invention, L 2 It is a double-stranded DNA oligonucleotide composed of 10 AT pairs.

[0085] In some embodiments of the present invention, L2 It is a double-stranded oligonucleotide of DNA / RNA hybridization, in which the DNA single strand is composed of 6 Ts and the RNA single strand is composed of 6 A.

[0086] In another embodiment of the present invention, L 2 The following scheme (4) is adopted: L 2 It is an oligopeptide residue consisting of 2-10 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid residues.

[0087] In some embodiments of the present invention, L 3 For a single key.

[0088] In some embodiments of the present invention, L 3 It is -N(H)-.

[0089] In some embodiments of the present invention, L 3 is a 4-8 membered saturated nitrogen-containing heterocyclic group, such as

[0090] Specifically, R 1 Selected from: H, C 1 -C 6 Alkyl, halogen, C 1 -C 6 Haloalkyl, C 1 -C 6 Alkoxy, C 1 -C 6 haloalkoxy; in some embodiments of the invention, R 1 For H.

[0091] The AR degrader provided by the present invention can effectively degrade AR and reduce the protein level of AR, and can be used for the prevention and treatment of diseases related to excessive AR, such as prostate cancer, which has gradually become one of the most common malignant tumors in men. Most treatment methods for prostate cancer are limited to androgen drugs or surgical castration therapy, but many prostate cancer patients inevitably still progress to castration-resistant prostate cancer (CRPC) after receiving androgen blockade therapy, among which the continuous activation of androgen receptor degrader (AR) signal is an important factor causing its deterioration. Degraders targeting AR are expected to treat patients with castration-resistant prostate cancer; breast cancer, AR receptors play an important role in the occurrence and development of breast cancer. About 70% of breast cancers express AR receptors, including some triple-negative breast cancers. The expression level of AR is closely related to the malignancy of triple-negative breast cancer. Abnormal and high expression of AR means that the AR expression level needs to be reduced to treat triple-negative breast cancer. Therefore, degraders targeting AR are expected to treat endocrine therapy resistance and AR-positive breast cancer patients; ovarian cancer, AR overexpression exists in most human ovarian cancers, androgen / AR signaling stimulates the proliferation, migration and invasion of ovarian cancer cells. Targeting AR has become an effective method for treating ovarian cancer, and AR-targeted degraders are expected to treat AR-positive ovarian cancer patients; pancreatic cancer, AR can be activated through non-androgen-derived pathways, and cytokine IL-6 is one of the AR activators. IL-6 is overexpressed in pancreatic cancer, upregulating the phosphorylation of STAT3 and MAPK, thereby activating AR function and promoting the survival and migration of ovarian tumor cells. AR overactivation plays an important role in pancreatic cancer, so AR-targeted degraders are expected to treat AR-positive pancreatic cancer patients; malignant tumors such as bladder cancer, kidney cancer, lung cancer and liver cancer; in addition, excessive androgens may cause many problems and lead to many "visual changes": such as acne, hirsutism, seborrhea, androgenic alopecia, precocious puberty or virilization, etc. AR-targeted degraders can be used as anti-androgen agents to treat the above-mentioned hyperandrogenism.

[0092] Specifically, ring A is a benzene ring or a 5-6-membered nitrogen-containing aromatic heterocycle, for example

[0093] In some embodiments of the invention, Ring A is

[0094] In some embodiments of the present invention, for

[0095] Specifically, R 2 Selected from: H, C 1 -C 6 Alkyl, halogen, C 1 -C6 Haloalkyl, C 1 -C 6 Alkoxy, C 1 -C 6 haloalkoxy; in some embodiments of the invention, R 2 For H.

[0096] Specifically, the B ring is selected from: Among them, R 2 '、R 2 ”、R 2 ”', having the above R 2 The definition of , a, b, c, d are independently selected from integers of 0-3, for example,

[0097] In some embodiments of the present invention, for

[0098] Specifically, R 3 Selected from: H, cyano, C 1 -C 6 Alkyl, halogen, C 1 -C 6 Haloalkyl, C 1 -C 6 Alkoxy, C 1 -C 6 haloalkoxy; in some embodiments of the invention, R 3 Halogen or cyano, such as F, Cl, Br, I, CN.

[0099] Specifically, the C ring is a benzene ring or a 5-6-membered nitrogen-containing aromatic heterocycle, for example

[0100] In some embodiments of the invention, Ring C is

[0101] In some embodiments of the present invention, for Among them, R 3 'With the above R 3 Definition of .

[0102] Specifically, Y 1 Selected from: single bond, -C(O)-, -C(S)-, -C(O)-N(C 0 -C 6 Alkyl)-, -C(S)-N(C 0 -C 6In some embodiments of the present invention, Y 1 is -C(O)-; in some embodiments of the present invention, Y 1 -C(O)-N(C 0 -C 6 In some embodiments of the present invention, Y 1 For a single key.

[0103] Specifically, Y 2 Selected from: single bond, -N(C 0 -C 6 alkyl), -C(O)-N(C 0 -C 6 Alkyl)-, -C(S)-N(C 0 -C 6 In some embodiments of the present invention, Y 2 -N(C 0 -C 6 In some embodiments of the present invention, Y 2 is a single bond; in some embodiments of the present invention, Y 2 -C(O)-N(C 0 -C 6 Alkyl)-, such as -C(O)-NH-.

[0104] Specifically, Y 3 Selected from: -O-, -S-, SO 2 In some embodiments of the present invention, Y 3 is O.

[0105] In some embodiments of the present invention, Part of

[0106]

[0107] In some embodiments of the present invention, the AR degrading agent has the following structure:

[0108]

[0109] Specifically, in formula II, Part of the configuration is

[0110] In some embodiments of the present invention, the AR degrading agent has the following structure:

[0111]

[0112] Specifically, in formula II and III, GRO, L 1and L 2 As mentioned above.

[0113] Specifically, in Formula II and III, L 1 Connected to the 3' end, 5' end or any base, sugar or phosphate backbone in the middle of GRO.

[0114] In some embodiments of the present invention, L 1 is a single bond or O, i.e. GRO is connected by a phosphate bond (i.e. ) and L 2 connect.

[0115] In some embodiments of the present invention, L 2 Adopting the above scheme (1) and / or (2), especially scheme (1), for example

[0116] More specifically, the AR degradation agent has the following structure:

[0117]

[0118] In one embodiment of the present invention, the AR degradation agent has the following structure:

[0119]

[0120] In other embodiments of the present invention, the AR degradation agent has the following structure:

[0121]

[0122]

[0123]

[0124]

[0125]

[0126] In the second aspect of the present invention, a method for preparing the AR degrading agent of the first aspect is provided, which comprises the step of coupling the small molecule ligand part with GROs via L.

[0127] In some embodiments of the present invention, the preparation method comprises the following steps: first preparing Connect it to a solid phase carrier, and then synthesize oligonucleotides through solid phase to obtain the target product;

[0128] L' is an optional suitable linking group;

[0129] R F A protected hydroxyl group.

[0130] Specifically, the solid phase synthesis adopts the phosphoramidite method, which includes four steps: deprotection, coupling, capping and oxidation, as described in Kosuri S, Church GM. Large-scale de novo DNA synthesis: technologies and applications. Nat Methods. 2014 May; 11(5): 499-507. Figure 2 shown.

[0131] Specifically, in solid phase synthesis, L' comprises (For example )

[0132] In some embodiments of the present invention, R F is -ODMTr.

[0133] In some embodiments of the present invention, the solid phase carrier is controlled pore glass beads (CPG), and the pore size of the CPG is determined according to the length of the synthesized oligonucleotide.

[0134] In some embodiments of the present invention, the preparation method comprises the following steps: first preparing Reaction with GROs; or,

[0135] First prepare GRO-L'-R F , and then linked to a small molecule ligand; or,

[0136] Prepare GRO-L"-R F 'and Then couple the two together;

[0137] Wherein, L', L", L'' are any suitable connecting groups, R F , R F '、R F " is an optional suitable reactive group.

[0138] In the third aspect of the present invention, a pharmaceutical composition comprises the AR degrading agent described in the first aspect or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate or deuterated compound thereof, and one or more pharmaceutically acceptable excipients.

[0139] Specifically, the pharmaceutically acceptable excipients may be selected from one or more of fillers, binders, lubricants, disintegrants, antioxidants, buffers, antibacterial agents, suspending agents, solubilizers, thickeners, stabilizers, preservatives, and the like.

[0140] Specifically, the pharmaceutical composition can be administered by any suitable route, such as gastrointestinal administration (e.g., oral, sublingual, rectal administration) or parenteral administration (e.g., intravenous, intramuscular, intranasal, intraocular, intracerebral, intravaginal, intraperitoneal, transdermal, subcutaneous, intradermal, respiratory tract administration, etc.).

[0141] Specifically, the pharmaceutical composition can be prepared into pharmaceutical preparations in the following forms: injection, syrup, elixir, suspension, powder, granule, tablet, capsule, lozenge, cream, ointment, lotion, gel, emulsion, etc.

[0142] When preparing injections, any commonly used carrier in the art can be used, such as water, ethanol, propylene glycol, ethoxylated isostearyl alcohol, polyethoxylated isostearyl alcohol, and fatty acid esters of polyethylene sorbitan, etc. In addition, commonly used solvents and buffers can also be added.

[0143] Specifically, the pharmaceutical composition is preferably in unit dosage form. In this form, the preparation is subdivided into unit doses containing appropriate amounts of active ingredients. The unit dosage form can be a capsule, tablet or any dosage form; in addition, the unit dosage form can also be a packaged preparation, such as tablets, capsules and powders packaged in vials or ampoules.

[0144] Specifically, in the pharmaceutical composition, the PROTAC can be used alone or in combination with other types of active ingredients.

[0145] Specifically, the amount of active ingredient in the unit dose formulation can be changed or adjusted from 0.1 mg to 1000 mg (e.g., 0.1, 1, 5, 10, 20, 40, 50, 100, 200, 400, 500, 1000 mg), depending on the specific application and efficacy of the active ingredient. If necessary, the composition may also include other suitable therapeutic agents.

[0146] In a fourth aspect of the present invention, a delivery system for an AR degrader is provided, which comprises the above-mentioned AR degrader and a carrier, and can deliver the AR degrader to target cells or target tissues without NCL expression on the cell surface.

[0147] Specifically, the above-mentioned vector can adopt any vector suitable for delivering nucleic acid drugs into target tissues or target cells, such as those disclosed in the prior art (see, for example, Wang Junfeng, Tan Manman, Wang Ying, etc. Research progress on modification and delivery of nucleic acid drugs [J]. Journal of Zhejiang University (Medical Edition), 2023, 52(04): 417-428. It is incorporated herein by reference), for example, viral vectors (such as lentivirus, adenovirus, adeno-associated virus vectors), non-viral vectors (such as lipid nanoparticles (LNP), polymer nanocarriers, inorganic nanocarriers, protein carriers, exosomes, etc.).

[0148] In some embodiments of the invention, the carrier is a lipid nanoparticle (LNP).

[0149] In the fifth aspect of the present invention, there is provided use of the AR degrading agent described in the first aspect or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate or deuterated compound thereof in the preparation of a medicament for preventing and / or treating AR-related diseases.

[0150] Specifically, the disease is a disease that can be beneficially prevented and / or treated by degrading AR, such as, but not limited to, tumors, autoimmune diseases, inflammatory diseases, diseases associated with pathogen infection, neurodegenerative diseases, cardiovascular diseases, metabolic diseases, fibrotic diseases, neuro-psychiatric disorders, etc.

[0151] Specifically, the tumor includes but is not limited to: lung cancer, malignant melanoma, brain tumor, tumor of digestive organs, uterine cancer, endometrial cancer, testicular cancer, palate cancer, pharyngeal cancer, tongue cancer, oral cancer, various sarcomas, osteosarcoma, blood system tumors, nervous system tumors, brain glioma, glioblastoma, skin cancer, skin appendage cancer and skin metastasis, medulloblastoma, blastoma, liposarcoma, neuroendocrine tumor, synovial cell sarcoma, gastrinoma, carcinoid tumor, mesothelioma, pancreatic islet cell carcinoma, schwannoma, meningioma, melanoma, acoustic neuroma, adenocarcinoma, lymphoid malignancies, epithelial squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer, squamous cell carcinoma, lung adenocarcinoma, peritoneal cancer, lung squamous cell carcinoma, hepatocellular carcinoma, gastric cancer, intestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, thyroid cancer, bladder cancer, breast cancer, metastatic breast cancer, colon cancer, rectal cancer, prostate cancer, salivary gland cancer, kidney cancer, vulvar cancer, anal cancer, penile cancer, esophageal cancer, biliary tract tumors, and head and neck cancer.

[0152] Specifically, the hematological malignancies include leukemia, lymphoma, and multiple myeloma (MM).

[0153] Specifically, the leukemia can be chronic lymphocytic leukemia (CLL) (e.g., B cell CLL, T cell CLL), chronic myeloid leukemia (CML) (e.g., B cell CML, T cell CML), acute lymphocytic leukemia (ALL) (e.g., B cell ALL, T cell ALL), acute myeloid leukemia (AML) (e.g., B cell AML, T cell AML), acute monocytic leukemia, etc.

[0154] Specifically, the lymphoma may be Hodgkin lymphoma (HL) (e.g., B cell HL, T cell HL) and non-Hodgkin lymphoma (NHL) (e.g., B cell NHL such as diffuse large cell lymphoma (DLCL) (e.g., diffuse large B cell lymphoma), follicular lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma (CLL / SLL), mantle cell lymphoma (MCL), marginal zone B cell lymphoma (e.g., mucosa-associated lymphoid tissue (MALT) lymphoma, nodular marginal zone B cell lymphoma, splenic marginal zone B cell lymphoma), primary mediastinal B cell lymphoma, Burkitt's lymphoma, lymphoplasmacytic lymphoma (i.e., Waldenstrom's macroglobulinemia ( macroglobulinemia), hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B lymphoblastic lymphoma and primary central nervous system (CNS) lymphomas and T-cell NHL, such as precursor T lymphoblastic lymphoma / leukemia, peripheral T-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g., mycosis fungoides, Sezary syndrome)), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy-type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma and anaplastic large cell lymphoma, NK / T-cell lymphoma, in particular diffuse large B-cell lymphoma (DLBCL).

[0155] In some embodiments of the present invention, the tumor is selected from: prostate cancer (such as castration-resistant prostate cancer), breast cancer (such as triple-negative breast cancer), bladder cancer, liver cancer, kidney cancer, lung cancer, endometrial cancer, ovarian cancer, pancreatic cancer, malignant melanoma, in particular prostate cancer.

[0156] Specifically, the autoimmune diseases include, but are not limited to, organ-specific autoimmune diseases, systemic lupus erythematosus, rheumatoid arthritis, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease, autoimmune hemolytic anemia, thyroid autoimmune disease, ulcerative colitis, etc.

[0157] Specifically, the inflammatory diseases include, but are not limited to, osteoarthritis, acute gout, multiple sclerosis, inflammatory bowel disease (such as Crohn's disease and ulcerative colitis), neuroinflammation, asthma, chronic obstructive airway disease, pneumonia, myositis, eczema, dermatitis, acne, cellulitis, occlusive disease, thrombosis, alopecia, nephritis, vasculitis, retinitis, uveitis, scleritis, sclerosing cholangitis, hypophysitis, thyroiditis, septic shock, systemic inflammatory response syndrome (SIRS), toxic shock syndrome, acute lung injury, ARDS (adult respiratory distress syndrome), acute renal failure, burns, pancreatitis (such as acute pancreatitis), postoperative syndrome, sarcoidosis, Herxheimer reaction, encephalitis, myelitis, meningitis and malaria, etc.

[0158] Specifically, the neurodegenerative diseases include, but are not limited to, Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), different types of spinocerebellar ataxia (SCA), Pick's disease, etc.

[0159] Specifically, the pathogen may be a microorganism, a parasite (protozoa, worms, etc.) or other agents.

[0160] Specifically, the microorganism can be selected from one or more of: viruses, chlamydia, rickettsia, mycoplasma, bacteria, spirochetes, fungi and the like.

[0161] In one embodiment of the present invention, the pathogen is a virus, for example, but not limited to, Adenoviridae (such as adenovirus), Herpesviridae (such as HSV1 (oral herpes), HSV2 (external genital herpes), VZV (varicella), EBV (Epstein-Barr virus), CMV (cytomegalovirus)), Poxviridae (such as smallpox virus, vaccinia virus), Papovaviridae (such as papillomavirus (HPV)), Parvoviridae (such as B19 virus), Hepadnaviridae (such as hepatitis B virus), Polyomaviridae (such as polyomavirus), Reoviridae (such as reovirus, rotavirus), Picornaviridae (such as enterovirus, foot-and-mouth disease virus), Caliciviridae (such as Norwalk virus, hepatitis E virus), Togaviridae (such as rubella virus), Arenaviridae (such as lymphocytic choriomeningitis virus), Retroviridae (HIV-1, HIV-2, HTLV-1), Flaviviridae ( Such as dengue virus, Zika virus, Japanese encephalitis virus, Chikungunya virus, yellow fever virus, hepatitis C virus, West Nile virus, etc.), Orthomyxoviridae (such as influenza virus (such as influenza A virus, influenza B virus, influenza C virus, etc.)), Paramyxoviridae (such as human parainfluenza virus (HPIV) type 1, HPV type 2, HPV type 3, HPV type 4, Sendai virus, mumps virus, measles virus, respiratory syncytial virus, Newcastle disease virus, etc.), Bunyaviridae (such as California encephalitis virus, Hantavirus), Rhabdoviridae (such as rabies virus), Filoviridae (such as Ebola virus, Marburg virus), Coronaviridae (such as HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, MERS-CoV, SARS-CoV-2, etc.), Astroviridae (such as astrovirus), Bornaviridae (such as Borna virus).

[0162] Specifically, the diseases related to the pathogen infection include but are not limited to: influenza, SARS, COVID-19, viral hepatitis (such as hepatitis A, hepatitis B, hepatitis C, hepatitis D, etc.), AIDS, rabies, dengue fever, Ebola virus disease, etc.

[0163] Specifically, the cardiovascular disease includes, but is not limited to: coronary heart disease, peripheral arterial disease, atherosclerosis, ischemic heart disease, ischemic cardiomyopathy, myocardial infarction, heart failure, angina pectoris, myocarditis, hypercholesterolemia, hypertension, ischemia-reperfusion injury, cerebrovascular ischemia (stroke), embolism (such as pulmonary embolism, renal embolism, hepatic embolism, gastrointestinal embolism or peripheral limb embolism) or myocardial ischemia, etc.

[0164] Specifically, the metabolic diseases include, but are not limited to: diabetes (such as type I diabetes, type II diabetes or gestational diabetes), obesity, insulin resistance, hyperinsulinemia, fatty liver (NASH or others), cachexia, hypercholesterolemia, gout, etc.

[0165] Specifically, the fibrotic diseases include, but are not limited to, myocardial fibrosis, pulmonary fibrosis, renal fibrosis, postoperative stenosis, keloid formation, cirrhosis, biliary cirrhosis, scleroderma, etc. Specifically, the neuro-psychiatric disorders include, but are not limited to, anxiety and depression.

[0166] In addition, excessive androgen receptors may also lead to diseases such as acne, hirsutism, seborrhea, androgen receptor alopecia, precocious puberty or virilization, etc. Therefore, the AR degrading agent of the present invention can be used to prevent and / or treat these diseases.

[0167] In some embodiments of the present invention, the disease is selected from: prostate cancer (such as castration-resistant prostate cancer), breast cancer (such as triple-negative breast cancer), bladder cancer, liver cancer, kidney cancer, lung cancer, endometrial cancer, ovarian cancer, pancreatic cancer, malignant melanoma, obesity, insulin resistance, hyperinsulinemia, type II diabetes, fatty liver (such as non-alcoholic fatty liver disease), polycystic ovary syndrome, hirsutism, acne, androgenic alopecia (such as female pattern hair loss), infertility, pregnancy loss, pregnancy complications (such as gestational diabetes, gestational hypertension), hyperandrogenemia, cardiovascular disease (such as hypertension, dyslipidemia), anxiety, depression, pelvic floor disease (such as pelvic floor dysfunction, pelvic pain syndrome).

[0168] In the sixth aspect of the present invention, a method for treating AR-related diseases is provided, comprising the step of administering to a subject in need thereof the AR degrader described in the first aspect of the present invention or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate or deuterated compound thereof, or the pharmaceutical composition described in the third aspect of the present invention, or the delivery system described in the fourth aspect of the present invention.

[0169] Specifically, the disease is as described in the fifth aspect of the present invention.

[0170] Specifically, the subject is a mammal, especially a human.

[0171] Specifically, the administration can be by any suitable route of administration, such as enteral administration (e.g., oral, sublingual, rectal) or parenteral administration (e.g., intravenous, intramuscular, intranasal, intraocular, intracerebral, intravaginal, intraperitoneal, transdermal, subcutaneous, intradermal, respiratory tract administration, etc.).

[0172] The present invention provides a PROTAC for degrading AR prepared by using GROs as a novel recruitment element of MDM2, and a preparation method and application thereof. The inventor discovered the new function of GROs in recruiting MDM2 through research, and used it to prepare a PROTAC targeting AR, which can effectively degrade AR and produce corresponding therapeutic effects (such as inhibiting the proliferation of tumor cells and promoting tumor cell apoptosis), and has very good application prospects and research value. BRIEF DESCRIPTION OF THE DRAWINGS

[0173] Figure 1 The result graph shows that NCL can bind to MDM2.

[0174] Figure 2 The results show that AS1411 is a GRO with stable G4 structural features.

[0175] Figure 3 The resulting image shows the faint G4 structural features of iSN04.

[0176] Figure 4 A shows the result that CRO cannot capture NCL and MDM2; Figure 4 B shows the results of AS1411 concentration-dependent capture of NCL and MDM2; Figure 4 C shows the result that high concentration of iSN04 can capture NCL, but iSN04 cannot capture MDM2.

[0177] Figure 5 A shows the result that CRO can neither bind to NCL nor recruit MDM2; Figure 5 B shows the result of AS1411 recruiting MDM2 in large quantities depending on its interaction with NCL; Figure 5 C shows that iSN04 can only bind to NCL but cannot recruit MDM2 dependent on the interaction with NCL.

[0178] Figure 6 A shows the result that AS1411 does not affect the interaction between NCL and MDM2; Figure 6 B shows the result of iSN04 blocking the binding of NCL to MDM2.

[0179] Figure 7 A shows the result that AS1411 can significantly bind to human prostate cancer cell 22Rv1. Figure 7 B shows the result that AS1411 can capture NCL and MDM2; Figure 7 C shows the result that silencing NCL can block the recruitment of MDM2 by AS1411.

[0180] Figure 8A shows a schematic diagram of GROs (such as AS1411) recruiting MDM2 depending on the interaction with NCL; Figure 8 B shows a schematic diagram of the possible mode of action of AR-targeting PROTAC formed by GROs (such as AS1411) as MDM2 recruitment elements.

[0181] Fig. 9 The graph shows the results of AR degradation by AS1411-ARL.

[0182] Fig.10 The figure shows the results that AS1411-ARL mediates the degradation of AR through the ubiquitin-proteasome pathway.

[0183] Fig.11 The figure shows the results of AS1411-ARL's dependence on MDM2 and NCL-mediated degradation of AR.

[0184] Fig.12 The results show that AS1411-ARL can inhibit the proliferation of human prostate cancer cell 22Rv1 and promote the apoptosis of 22Rv1 cells. DETAILED DESCRIPTION

[0185] Unless otherwise defined, all scientific and technical terms used in the present invention have the same meanings as commonly understood by one of ordinary skill in the art to which the present invention relates.

[0186] The term "alkyl" refers to a straight or branched hydrocarbon chain radical that does not contain an unsaturated bond, and the hydrocarbon chain radical is connected to the rest of the molecule by a single bond. A typical alkyl group contains 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, preferably 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl, isohexyl, etc. In the present invention, C 0 Alkyl refers to H, i.e. C 0-10 Alkyl (or C 0 -C 10 Alkyl) includes H and C 1-10 Alkyl (or C 1 -C 10 alkyl).

[0187] The term "alkylene" refers to a hydrocarbon group (divalent alkyl) formed by losing two hydrogen atoms from an alkane molecule, which can be a straight chain or branched chain and is connected to the rest of the molecule by a single bond. In this context, a typical alkylene group has 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, preferably 1 to 6 carbon atoms, such as a methylene group (-CH 2 -), ethylene, propylene, butylene, etc. In the present invention, C0 Alkylene refers to a single bond, i.e. C 0-10 Alkylene (or C 0 -C 10 Alkylene) includes single bonds and C 1-10 Alkylene (or C 1 -C 10 alkylene).

[0188] The term "cycloalkyl" refers to alicyclic hydrocarbons, such as those containing 1 to 4 single rings and / or condensed rings, 3-18 carbon atoms, preferably 3-10 (e.g. 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or adamantyl, etc.

[0189] The term "alkoxy" refers to a substituent formed by replacing the hydrogen in a hydroxy group with an alkyl group, such as an alkoxy group containing 1 to 10 carbon atoms, for example, methoxy, ethoxy, propoxy, butoxy, and the like.

[0190] The term "alkylamino" refers to an amino group (-NH 2 ) is replaced by an alkyl group, such as an alkylamino group containing 1 to 10 carbon atoms, for example

[0191] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0192] The term "haloalkyl" refers to a group formed by replacing one or more hydrogen atoms in an alkyl group with a halogen atom (such as fluorine, chlorine, bromine or iodine), such as -CHF 2 、-CH 2 F, -CF 3 、-CH 2 -CF 3 、-CH 2 CH 2 -CF 3 、-CH 2 CH 2 CH 2 -CF 3 .

[0193] The term "aryl" refers to a monocyclic or polycyclic free radical, including a polycyclic free radical containing a monocyclic aromatic group and / or a condensed aromatic group, such as a free radical containing 1-3 monocyclic or condensed rings and 6-18 (e.g., 6, 8, 10, 12, 14, 16, 18) carbon ring atoms. 6 -C 12 The aryl group refers to an aryl group containing 6 to 12 carbon ring atoms, such as phenyl, naphthyl, biphenyl, indenyl and the like.

[0194] The term "heterocyclyl" refers to a 3- to 18-membered non-aromatic ring group containing 2 to 17 carbon atoms and 1 to 10 heteroatoms. The heterocyclyl group can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which can contain fused, spirocyclic or bridged ring systems. The heterocyclyl group can be partially saturated (heteroaryl) or fully saturated (heterocycloalkyl). Suitable heteroaryl groups in the compounds of the present invention contain 1, 2 or 3 heteroatoms selected from N, O, S and P atoms, and include, for example, coumarin, including 8-coumarin, quinolyl, including 8-quinolyl, isoquinolyl, pyridinyl, pyrazinyl, pyrazolyl, pyrimidinyl, furanyl, pyrrolyl, thienyl, thiazolyl, isothiazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, imidazolyl, indolyl, isoindolyl, indazolyl, indolizinyl, phthalazinyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, pyridazinyl, triazinyl, cinnolinyl, benzimidazolyl, benzofuranyl, benzofurazanyl, benzothienyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl and furopyridinyl. Suitable heterocycloalkyl groups in the compounds of the present invention contain 1, 2 or 3 heteroatoms selected from N, O or S atoms, and include, for example, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, oxathiolanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxirane, thiirane, azepine, oxazepine, diazepine, In the present invention, for the optionally substituted heterocyclic group, the substituted position can be any suitable carbon atom or heteroatom, for example, The substitution position of R can be on any suitable carbon atom or nitrogen atom, which can be, for example

[0195] The term "pharmaceutically acceptable salts" includes acid addition salts and base addition salts.

[0196] The term "acid addition salt" includes, but is not limited to, salts from inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydroiodic and phosphonic acids, and salts from organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, alkanedioic acids, aromatic acids and aliphatic and aromatic sulfonic acids. Therefore, these salts include but are not limited to sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, hydrochloride, hydrobromide, iodate, acetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, maleate, tartrate and mesylate, also comprising amino acid salts such as arginate, gluconate, galacturonate etc. Acid addition salts can be prepared by contacting the free alkali form with a sufficient amount of the required acid to form a salt in a conventional manner. The free alkali form can be regenerated by contacting the salt form with an alkali, and the free alkali is separated in a conventional manner.

[0197] The term "base addition salt" refers to a salt formed with a metal or amine, such as an alkali metal and alkaline earth metal hydroxide, or with an organic amine. Examples of metals used as cations include, but are not limited to, sodium, potassium, magnesium, and calcium. Examples of suitable amines include, but are not limited to, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine (ethane-1,2-diamine), N-methylglucamine, and procaine. Base addition salts can be prepared by contacting the free acid form with a sufficient amount of the desired base in a conventional manner to form a salt. The free acid form can be regenerated by contacting the salt form with an acid and isolating the free acid in a conventional manner.

[0198] In the present invention, "D" refers to deuterium; "substituted by deuterium" means replacing one or more hydrogen atoms with a corresponding number of deuterium atoms.

[0199] It should be recognized that, depending on the source of the chemical materials used in the synthesis, there is some variation in the natural isotopic abundance in the synthesized compounds. Therefore, the compounds of the present invention will inherently contain small amounts of deuterated isotopologues. Despite this variation, the concentration of such naturally abundant stable hydrogen and carbon isotopes is still very low and insignificant compared to the degree of stable isotopic substitution of the compounds of the present invention. See, for example, Wada, E et al., Seikagaku, 1994, 66: 15; Gannes, LZ et al., Comp Biochem Physiol Mol Integr Physiol, 1998, 119: 725.

[0200] In the compounds of the present invention, any atom not designated as deuterium is present at its natural isotopic abundance. Unless otherwise specified, when a position is specifically designated as "H" or "hydrogen", the position is understood to have hydrogen according to its natural abundance isotopic composition. Similarly, unless otherwise specified, when a position is specifically designated as "D" or "deuterium", the position is understood to have deuterium at an abundance of at least 3000 times greater than the natural abundance of deuterium (which is 0.015%) (i.e., at least 45% deuterium incorporation).

[0201] As used herein, the term "isotopic enrichment factor" refers to the ratio between the isotopic abundance and the natural abundance of a particular isotope.

[0202] In other embodiments, the compounds of the invention have an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation).

[0203] The term "isotopologue" refers to species wherein the chemical structure differs from a specific compound of the present invention only in its isotopic composition.

[0204] The term "stereoisomer" includes the presence of enantiomers, diastereomers and geometric isomers. Some compounds of the present invention have cyclic hydrocarbon groups that can be substituted on more than one carbon atom, in which case all geometric forms, including cis and trans, and mixtures thereof, are within the scope of the present invention.

[0205] The term "solvate" refers to a physical association of a compound of the invention with one or more solvent molecules. The physical association includes various degrees of ionic and covalent bonding, including hydrogen bonding. In some cases, the solvate can be isolated, for example when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. Solvates include solution phases and separable solvates. Representative solvates include ethanolates, methanolates, and the like.

[0206] The term "prodrug" refers to a form of the compound of formula I which is suitable for administration to patients without undue toxicity, irritation, allergic response, etc. and is effective for its intended use, including acetal, ester and zwitterion forms. The prodrug is transformed in vivo, such as by hydrolysis in the blood, to yield the parent compound.

[0207] In the present invention, the term "oligonucleotide" is composed of 30 or less (e.g., 5, 10, 15, 20, 25, 30) nucleotides (deoxyribonucleotides and / or ribonucleotides), which are single-stranded or double-stranded, especially single-stranded. The oligonucleotide may include chemical modification, nucleic acid unit replacement or connection functional groups; wherein the chemical modification is that at least one base is modified, and the chemical modification includes at least one of phosphorylation, methylation, amination, sulfhydrylation, isotopization, thiophosphate backbone modification, methoxy modification, and fluorination modification; the nucleic acid unit is replaced by at least one nucleic acid unit replaced by LNA, UNA or GNA; the functional group includes at least one of a fluorescent group, a radioactive group, biotin, digoxin, a nanoluminescent material, a nucleic acid substance or an enzyme marker.

[0208] In the present invention, for deoxyribonucleotides, A represents deoxyadenine nucleotide, T represents deoxythymidine, C represents deoxycytidine, and G represents deoxyguanosine; for ribonucleotides, A represents adenine ribonucleoside, G represents guanine ribonucleoside, C represents cytosine ribonucleoside, and U represents uracil ribonucleoside.

[0209] In the present invention, the term "oligopeptide" refers to a peptide composed of 2-10 (e.g., 2, 3, 4, 5, 6, 8, 10) amino acids, "polypeptide" contains 11-50 (e.g., 15, 20, 30, 40, 50) amino acids, and "protein" contains more than 50 amino acids.

[0210] The term "patient" or "subject" and the like are used interchangeably herein and refer to any animal or cell thereof treated according to the methods described herein, whether in vitro or in situ. Specifically, the aforementioned animal includes mammals, e.g., rats, mice, guinea pigs, rabbits, dogs, monkeys or humans, particularly humans.

[0211] The term "treating" refers to preventing, curing, reversing, attenuating, alleviating, minimizing, inhibiting, suppressing and / or halting one or more clinical symptoms of a disease after onset of the disease.

[0212] The term "prevent" refers to avoiding, minimizing or making the onset or development of a disease difficult by treating it before it occurs.

[0213] The term "tumor" refers to an abnormal mass of tissue, wherein the growth of the mass exceeds the growth of normal tissue and is not coordinated with the growth of normal tissue. Tumors can be "benign" or "malignant", depending on the following characteristics: degree of cell differentiation (including morphology and function), growth rate, local invasion and metastasis. "Benign tumors" are usually well-differentiated, characterized by slower growth than malignant tumors, and remain confined to the site of origin. In addition, benign tumors do not have the ability to infiltrate, invade or metastasize to distant sites. In some cases, some "benign" tumors may later lead to malignant tumors, which may be caused by additional genetic changes in a subpopulation of the neoplastic cells of the tumor, and these tumors are called "precancerous tumors". "Malignant tumors" are usually poorly differentiated (anaplastic) and have a characteristic rapid growth, accompanied by progressive infiltration, invasion and destruction of surrounding tissues. In addition, malignant tumors usually have the ability to metastasize to distant sites.

[0214] The term "cancer" refers to a malignant tumor (Stedman's Medical Dictionary, 25th ed.; Hensyled.; Williams & Wilkins: Philadelphia, 1990).

[0215] The term "autoimmune disease" refers to a disease caused by the body's immune response to its own antigens, resulting in damage to its own tissues.

[0216] The term "inflammation" is the body's defense response to stimulation, manifested as redness, swelling, heat, pain, and dysfunction, etc. It can be infectious inflammation caused by infection, or non-infectious inflammation not caused by infection, such as inflammation caused by immune response (such as various types of hypersensitivity reactions, inflammation caused by some autoimmune diseases). The term "inflammatory disease" refers to a disease with inflammation.

[0217] The term "disease associated with pathogen infection" mainly refers to diseases caused by pathogen infection, including symptoms of body damage caused by pathogen invasion and manifestations of infection response. The pathogen can be a microorganism (such as virus, chlamydia, rickettsia, mycoplasma, bacteria, spirochetes, fungi, etc.), a parasite (protozoa, worms, etc.) or other vectors. In particular, the pathogen is a virus, for example, but not limited to, adenoviridae (such as adenovirus), herpesviridae (such as HSV1 (oral herpes), HSV2 (external genital herpes), VZV (varicella), EBV (Epstein-Barr virus), CMV (cytomegalovirus)), poxviridae (such as smallpox virus, cowpox virus), papovarus, leukoencephalitis, leukemia ... Viridae (such as papillomavirus (HPV)), Parvoviridae (such as B19 virus), Hepadnaviridae (such as hepatitis B virus), Polyomaviridae (such as polyomavirus), Reoviridae (such as reovirus, rotavirus), Picornaviridae (such as enterovirus, foot-and-mouth disease virus), Caliciviridae (such as Norwalk virus, hepatitis E virus), Togaviridae (such as rubella virus), Arenaviridae (such as lymphocytic choriomeningitis virus), Retroviridae (such as HIV), -1, HIV-2, HTLV-1), Flaviviridae (such as dengue virus, Zika virus, Japanese encephalitis virus, Chikungunya virus, yellow fever virus, hepatitis C virus, West Nile virus, etc.), Orthomyxoviridae (such as influenza virus (such as influenza A virus, influenza B virus, influenza C virus, etc.)), Paramyxoviridae (such as human parainfluenza virus type 1 (HPIV), HPV type 2, HPV type 3, HPV type 4, Sendai virus, mumps virus, measles virus, respiratory syncytial virus, Newcastle disease virus, etc.), Bunyaviridae (such as California encephalitis virus, Hantavirus), Rhabdoviridae (such as rabies virus), Filoviridae (such as Ebola virus, Marburg virus), Coronaviridae (such as HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, MERS-CoV, SARS-CoV-2, etc.), Astroviridae (such as Astrovirus), Bornaviridae (such as Borna virus).

[0218] The term "cardiovascular disease" refers to a group of diseases involving the heart or blood vessels.

[0219] The term "metabolic disease" refers to diseases caused by the accumulation or deficiency of certain metabolic substances such as sugar, fat, protein, purine, calcium and copper when the biochemical processes in the body are disturbed.

[0220] The term "fibrosis" refers to the pathological process in which inflammation leads to necrosis of organ parenchymal cells, abnormal increase and excessive deposition of extracellular matrix in tissues. In severe cases, it causes destruction of tissue structure and organ sclerosis.

[0221]

[00136] Various publications, patents, and published patent specifications are cited herein, the disclosures of which are incorporated by reference in their entireties.

[0222] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. 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.

[0223] The sequences involved in the embodiments are shown in the following table:

[0224] Table 1 Sequence name and number

[0225] name sequence serial number AS1411 5′-GGTGGTGGTGGTTGTGGTGGTGGTGG-3′ SEQ ID NO:1 iSN04 5'-AGATTAGGGTGAGGGTGA-3' SEQ ID NO:2 CRO 5′-CCTCCTCCTCCTTCCTCCTCCTCC-3′ SEQ ID NO:3 NC 5'-GGAATTCCCGGTGCGCCGATCGCCGGATATAACTT-3' SEQ ID NO:4 sI 5'-UUCUCCGAACGUGUCACGUTT-3' SEQ ID NO:5 siNCL 5'-GGAUGACGACGACGACGAAGATT-3' SEQ ID NO:6 siMDM2 5'-GCUUGGCCUACAGUCAUCUTT-3' SEQ ID NO:7

[0226] Example 1: NCL can bind to MDM2

[0227] (1) Co-immunoprecipitation (Co-IP) is a classic method for studying protein interactions based on the specific interaction between antibodies and antigens. Hep3B liver cancer cells were lysed with IP lysis buffer (purchased from Thermo Scientific, Catalog No. 87788) and incubated with NCL antibodies at 4°C overnight. Protein A / G magnetic beads (purchased from Thermo Scientific, Catalog No. 88802) were added and incubated at room temperature for 2 hours with rotation. NCL and its interacting protein complex bound to the magnetic beads were washed with IP lysis buffer (purchased from ThermoScientific, Catalog No. 87788), and SDS-PAGE protein loading buffer (purchased from Beyotime, Catalog No. P0015) was added and heated to 100°C for 10 minutes. The magnetic beads were adsorbed with a magnetic stand and the supernatant was transferred to a new tube. Then, a western blot was performed to detect whether NCL binds to MDM2. The steps of the immunoblotting experiment are as follows: the protein samples were separated by SDS-PAGE electrophoresis, and the separated proteins were transferred to a PVDF membrane. After blocking with TBST buffer containing 5% skim milk at room temperature for 1 hour, the membrane was incubated with the primary antibody of MDM2 (purchased from proteintech, catalog number 27883-1-AP) or the primary antibody of NCL (purchased from Cell Signaling Technology, catalog number 14574S) at 4°C overnight, and then washed with TBST. The membrane was incubated with HRP-labeled secondary antibody (purchased from Abotek, catalog number AS014) at room temperature for 1 hour, and the protein band blot was visualized using an enhanced chemiluminescence detection kit (purchased from Abotek, catalog number RM00021P). Figure 1The results of A indicate that NCL can bind to MDM2.

[0228] (2) Mix 2 μg / mL recombinant human NCL (rhNCL, purchased from ACROBiosystems, NUL-H5253) and 2 μg / mL recombinant human MDM2 (rhMDM2, purchased from R&D Systems, E3-202-050) and incubate at 4°C for 7 hours. Then add the antibody against NCL and continue incubation at 4°C overnight. Add Protein A / G magnetic beads (purchased from Thermo Scientific, catalog number 88802) and incubate at room temperature for 1.5 hours with rotation. Wash the magnetic beads three times with TBST buffer, add SDS-PAGE protein loading buffer (purchased from Bio-Tech, catalog number P0015), and heat to 100°C for 10 minutes. Use a magnetic stand to adsorb the magnetic beads and take the supernatant to a new tube. Then perform an immunoblotting experiment to detect whether NCL binds to MDM2. Figure 1 The results in B indicate that NCL can bind to MDM2.

[0229] Example 2: AS1411 is a GRO with stable G4 structural features

[0230] GROs contain one or more GGT motifs and have stable G4 structural features. GROs are guanine-rich oligonucleotides that can specifically bind to NCL. The GROs described in the present invention are one of the DNA aptamers AS1411 and AS1411 derivatives / analogs, all of which have the above-mentioned stable G4 structural features and can specifically bind to NCL. NCL is a multifunctional shuttle protein that shuttles between the nucleus and cytoplasm of the cell. NCL is expressed in a variety of diseased cells (such as tumors, etc.) and is specifically transferred to the cell surface, affecting multiple processes such as cell proliferation, apoptosis, and metastasis. However, NCL is not expressed on the surface of normal cells, so that NCL is used as a surface molecular marker and potential therapeutic target for diseased cells (such as tumors, etc.). NCL on the membrane surface of diseased cells (such as tumors, etc.) can also act as a receptor for a variety of ligand molecules, mediating the entry of ligand molecules into diseased cells (such as tumors, etc.).

[0231] (1) AS1411 and iSN04 are both oligonucleotides that specifically bind to NCL. We tested whether AS1411 and iSN04 have stable G4 structural features. The experimental process is as follows: 5 μM NC (negative control, SEQ ID NO: 4), AS1411 or iSN04 were added to water to a total volume of 100 μL, heated at 95°C for 7 minutes, and immediately incubated on ice for 3 minutes. Then, 2 μL N-methyl mesoporphyrin IX (NMM) (purchased from MCE, product number HY-133821) was added to make the final concentration 1 μM, and incubated at room temperature in the dark for 1 hour. The fluorescence intensity was detected using an ELISA reader with an emission wavelength of 550-700 nm and an excitation wavelength of 399. The results are shown in Figure 2. Figure 2 As shown in Figure 2, no G4 structural features were detected in iSN04, but AS1411 had stable G4 structural features (fluorescence signal intensity exceeded 4000).

[0232] (2) Guanine-rich DNA sequences can form more obvious G4 structural features in the presence of metal ions (usually sodium and potassium ions). 5 μM NC (negative control, SEQ ID NO: 4), AS1411 or iSN04 were added to 100 mM K + The total volume of the solution was 100 μL. After thorough mixing, the mixture was heated at 95°C for 7 minutes, immediately placed on ice for incubation for 3 minutes, and then 2 μL of N-methyl mesoporphyrin IX (NMM) (purchased from MCE, item number HY-133821) was added to make the final concentration 1 μM. The mixture was incubated at room temperature in the dark for 1 hour. The fluorescence intensity was detected using an ELISA reader, with an emission wavelength of 550-700 nm and an excitation wavelength of 399. The results are shown in Figure 2. Figure 3 iSN04 has weak G4 structural features (fluorescence signal intensity 1000-2000), but AS1411 has more significant G4 structural features (fluorescence signal intensity over 10000).

[0233] The above results show that iSN04 can only + In the presence of K, a weak G4 structure can be formed. + The G4 structural features cannot be formed in the absence of K, but AS1411 + Stable G4 structural characteristics can be formed in the presence of.

[0234] Example 3: AS1411 recruits MDM2 via interaction with NCL

[0235] (1) The process of the pull-down experiment is to fix a substance of known identity (bait) on a carrier and use it to capture the binding protein (prey) and the protein that interacts with the binding protein from a complex mixture. This experiment is used to prove the possible interaction between the bait and a certain protein or protein complex. Hep3B liver cancer cells were lysed with IP lysis buffer (purchased from ThermoScientific, catalog number 87788), and then incubated with different concentrations (0nM, 200nM, 500nM, 1μM, 5μM, 10μM, 20μM) of 5'-end biotin-labeled CRO (cytosine-rich oligonucleotides, negative control), AS1411 or iSN04 at 4℃ for 6 hours, followed by the addition of streptavidin agarose gel beads (purchased from cytiva, catalog number 17511301) and continued incubation at 4℃ overnight. After washing the biotin-labeled CRO, AS1411 or iSN04 bound to the gel beads and the captured proteins multiple times with IP lysis buffer (purchased from Thermo Scientific, Catalog No. 87788), SDS-PAGE protein loading buffer (purchased from Beyotime, Catalog No. P0015) was added and heated to 100°C for 10 minutes. After centrifugation, the supernatant was taken, which was the pull-down product. The pull-down product was detected by immunoblotting. Figure 4 The results of A showed that CRO could capture neither NCL nor MDM2; Figure 4 The results in B show that AS1411 can capture NCL and MDM2 in a concentration-dependent manner; Figure 4 The results of C showed that only high concentrations of iSN04 could capture NCL, but iSN04 could not capture MDM2 within the concentration range used.

[0236] (2) 6 μg / mL recombinant human NCL and 6 μg / mL recombinant human MDM2 were mixed and incubated at 4°C for 7 hours, followed by the addition of 400 nM 5'-end biotin-labeled CRO, AS1411 or iSN04, incubated at 4°C for 6 hours, and then added with streptavidin agarose gel beads (purchased from cytiva, catalog number 17511301) and continued to incubate at 4°C overnight. The gel beads were washed 4 times with TBST buffer, and SDS-PAGE protein loading buffer (purchased from Biyuntian, catalog number P0015) was added and heated to 100°C for 10 minutes. The supernatant was collected after centrifugation. The pull-down product was then detected by immunoblotting. Figure 5 The results of A indicate that CRO can neither bind to NCL nor recruit MDM2. Figure 5 The results in B indicate that AS1411 recruits MDM2 in large quantities depending on its interaction with NCL. Figure 5 The results in C indicate that iSN04 can only bind to NCL but cannot recruit MDM2 dependently on the interaction with NCL.

[0237] (3) Hep3B liver cancer cells were lysed with IP lysis buffer (purchased from Thermo Scientific, catalog number 87788) and incubated with different concentrations (0 μM, 1 μM, 5 μM, 10 μM, 20 μM) of AS1411 or iSN04 at 4°C for 6 hours, followed by the addition of 5 μL of NCL antibody and continued incubation at 4°C overnight. Protein A / G magnetic beads (purchased from Thermo Scientific, catalog number 88802) were added and incubated at room temperature for 1.5 hours with rotation. NCL bound to the magnetic beads and the captured proteins were washed with IP lysis buffer (purchased from Thermo Scientific, catalog number 87788), and SDS-PAGE protein loading buffer (purchased from Beyotime, catalog number P0015) was added and heated to 100°C for 10 minutes. The magnetic beads were adsorbed with a magnetic stand and the supernatant was transferred to a new tube. Then, immunoblotting (western blot) was performed to detect whether NCL binds to MDM2 and whether the binding is affected by AS1411 or iSN04. Figure 6 The results of A indicate that AS1411 does not affect the interaction between NCL and MDM2. The same experiment was performed using derivatives / analogues of AS1411, and the experimental results were similar to those of AS1411. However, Figure 6 The results in B indicate that iSN04 blocks the binding of NCL to MDM2.

[0238] (4) Since NCL is highly expressed in a variety of tumor cells (including prostate cancer cells) and specifically distributed on the surface of tumor cells, we tested the binding ability of AS1411 to human prostate cancer cells 22Rv1. 22Rv1 cells were incubated with 500 nM Cy5-labeled CRO and AS1411 for 2 hours. The binding ability of each of them to 22Rv1 cells was detected by flow cytometry. The results are shown in Figure 2. Figure 7 As shown in A. The results show that AS1411 can significantly bind to 22Rv1 cells.

[0239] (5) Human prostate cancer cells 22Rv1 were incubated with 1 μM biotin-labeled negative control CRO or AS1411 for 12 hours. After the cells were lysed, the supernatant was incubated with streptavidin agarose gel beads (purchased from cytiva, catalog number 17511301) at 4°C overnight. The biotin-labeled CRO or AS411 bound to the gel beads and the captured proteins were washed several times with IP lysis buffer (purchased from Thermo Scientific, catalog number 87788), and then SDS-PAGE protein loading buffer (purchased from Biotech, catalog number P0015) was added and heated to 100°C for 10 minutes. After centrifugation, the supernatant was taken, which was the pull-down product. The pull-down product was detected by immunoblotting. The results are shown in Figure 7 As shown in B, AS1411 can capture NCL and MDM2.

[0240] (6) Negative control siRNA (siNC, SEQ ID NO: 5) or NCL siRNA (siNCL, SEQ ID NO: 6) was transfected into human prostate cancer cells 22Rv1. After 48 hours, the cells were incubated with 1 μM biotin-labeled AS1411 for 12 hours, the cells were lysed, and the supernatant was incubated with streptavidin agarose beads at 4°C overnight. The biotin-labeled AS1411 bound to the gel beads and the captured proteins were washed several times with IP lysis buffer (purchased from Thermo Scientific, catalog number 87788), and then SDS-PAGE protein loading buffer (purchased from Bio-Tech, catalog number P0015) was added and heated to 100°C for 10 minutes. After centrifugation, the supernatant, i.e., the pull-down product, was taken and the pull-down product was detected by immunoblotting. The results are shown in Figure 7 C, Silencing of NCL can block the recruitment of MDM2 by AS1411.

[0241] Example 4: Preparation of PROTAC molecules targeting AR based on GROs

[0242] We simulated the 3D structures of AS1411 and NCL-MDM2 complex using Discovery Studio and AlphaFold2, respectively, and used HDOCK to predict the conformation of the interaction between AS1411 and NCL-MDM2 complex. The results showed that GROs (such as AS1411) rely on the interaction with NCL to recruit MDM2. The possible modes of action are as follows: Figure 8 As shown in A; GROs (such as AS1411) as MDM2 recruitment elements to form a possible mode of action of AR-targeting PROTAC is as follows Figure 8 As shown in B.

[0243] In this example, AS1411, a typical representative of GROs, was used as the E3 ligase recruitment element, and ARL (L-7) was used as an example of the target protein ligand to prepare a PROTAC that degrades AR (hereinafter referred to as AS1411-ARL).

[0244] The preparation steps are as follows:

[0245] (1) Synthesis of Compound L-1

[0246]

[0247] To a solution of compound 2 (10.1 g, 46.93 mmol) in DMF (100 mL) was added NaH (2.25 g, 56.31 mmol) and stirred at -10°C for 1 hour. Compound 1 (7.3 g, 46.93 mmol) was then added to the above solution and stirred at -10°C for 1 hour. The mixture was poured into ice / water and filtered. The residue was dried in vacuo to obtain the target product L-1 (white solid, 12 g, 34.2 mmol, yield: 72.9%).

[0248] LCMS: m / z=351.0 [M+H] + t R =1.18min.Purity:97.8%(254nm).

[0249] (2) Synthesis of Compound L-2

[0250]

[0251] Chloroacetyl (10.74 g, 136.81 mmol) was added to a methanol (300 mL) solution at 0°C and stirred at room temperature for 20 minutes. Compound L-1 (12.0 g, 34.20 mmol) was added to the above solution and stirred for 3 hours. The mixture was concentrated and dried to obtain the target product L-2 (white solid, 10 g, yield: 100%).

[0252] LCMS: m / z = 251.0 [M+H] + t R =1.13min.Purity:94%(254nm).

[0253] (3) Synthesis of Compound L-3

[0254]

[0255] Oxalyl chloride (914 mg, 7.2 mmol) was added to a solution of compound 3 (571 mg, 3.6 mmol) in DCM (20 mL) and stirred for 1 hour. The mixture was concentrated to dryness, and then a solution of compound L-2 (903 mg, 3.6 mmol) in DCM (20 mL) and TEA (1091 mg, 10.8 mmol) was added and stirred for 2 hours. The mixture was concentrated to dryness and purified by silica gel column chromatography, eluting with MeOH:DCM = 0-5% to obtain the target product L-3 (white solid, 550 mg, 1.41 mmol, yield: 39%).

[0256] LCMS: m / z=391[M+H] + t R=1.59min.Purity:97%(254nm).

[0257] (4) Synthesis of Compound L-4

[0258]

[0259] Compound L-3 (800 mg, 2.04 mmol), compound 4 (557 mg, 93.07 mmol) and DIPEA (793 mg, 6.13 mmol) were dissolved in anhydrous DMSO (10 mL) and stirred at 100 ° C for 4 hours. The mixture was diluted with ethyl acetate (20 mL) and washed with brine (15 mL * 3). The organic layer was concentrated and dried, and purified by silica gel column chromatography, washed with ethyl acetate: petroleum ether = 0-50%, to obtain the target product L-4 (light yellow solid, 450 mg, 0.9 mmol, yield: 44%).

[0260] LCMS: m / z = 500.0 [M+H] + t R =1.43min.Purity:86%(254nm).

[0261] (5) Synthesis of Compound L-5

[0262]

[0263] To a solution of compound L-4 (500 mg, 1.0 mmol) in 1,4-dioxane (20 mL) and water (6 mL) was added LiOH. 2 O (210 mg, 5 mmol), stirred at room temperature for 3 hours. The mixture was concentrated to dryness, and then the residue was adjusted to pH = 5 and filtered to obtain the target product L-5 (white solid, 490 mg, yield: 100%).

[0264] LCMS: m / z = 486.0 [M+H] + t R =1.36min.Purity:100%(254nm).

[0265] (6) Synthesis of Compound L-6

[0266]

[0267] DIPEA (532 mg, 4.12 mmol) was added to a DCM (15 mL) solution of compound L-5 (500 mg, 1.03 mmol). The mixture was stirred at 25 °C for 10 minutes under an argon atmosphere. Then 2,2,2-trifluoroacetic acid pentafluorophenyl ester (577 mg, 2.06 mmol) was slowly added at low temperature. The mixture was stirred at 25 °C overnight under an argon atmosphere. LCMS showed that the reaction was complete. The mixture was then diluted with DCM (100 mL), washed with water (50 mL*3), brine (25 mL), and washed with anhydrous Na 2 SO 4 After drying, the organic layer was concentrated under reduced pressure after filtration. The residue was purified by silica gel column chromatography using MeOH:DCM = 15% as eluent to obtain the target product L-6 (white solid, 420 mg, yield: 62.6%).

[0268] LCMS: m / z=652[M+H] + t R =1.985min.Purity:79.23%(254nm).

[0269] (7) Synthesis of Compound L-7

[0270]

[0271] DIPEA (416 mg, 3.22 mmol) was added to a solution of compound L-6 (420 mg, 0.644 mmol) in ACN (15 mL). The mixture was stirred at 25 ° C for 10 minutes under an argon atmosphere. Then (3S, 5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)pyrrolidin-3-ol (405 mg, 0.966 mmol) was added. The mixture was stirred at 25 ° C overnight under an argon atmosphere. LCMS showed that the reaction was complete. The mixture was then diluted with DCM (100 mL), washed with water (50 mL*3), brine (25 mL), and washed with anhydrous Na 2 SO 4 After drying, the organic layer was concentrated under reduced pressure after filtration. The residue was purified by silica gel column chromatography using MeOH:DCM=15% as eluent to obtain the target product L-7 (white solid, 460 mg, yield: 80.1%).

[0272] LCMS: m / z=887[M+H] + t R =1.916min.Purity:100%(214nm).

[0273] (8) Synthesis of Compound L-8

[0274]

[0275] DMAP (253 mg, 2.07 mmol) was added to a solution of compound L-7 (460 mg, 0.518 mmol) in DCM (15 mL). The mixture was stirred at 25 ° C for 10 minutes under an argon atmosphere. Then dihydrofuran-2,5-dione (78 mg, 0.778 mmol) was added. The mixture was stirred overnight at 25 ° C under an argon atmosphere. LCMS showed that the reaction was complete. The mixture was concentrated and diluted with water (50 mL), and the pH was slowly adjusted to 7 with HCl (1 M) at low temperature. The mixture was extracted with EA (100 mL * 4), concentrated and purified by reverse phase chromatography [ACN / water (TEA 0.02%): 5%-30%] to obtain the target product L-8 (white solid, 470 mg, yield: 92%).

[0276] LCMS: m / z = 987.0 [M+H] + t R =1.695min.Purity:100%(214nm).

[0277] 1 H NMR (400MHz, DMSO) δ8.63–8.39(m,1H),7.85(m,1H),7.71(m,1H),7.41(m,2H),7.35–7.25(m,4H),7.2 4–7.08(m,6H),6.95–6.76(m,5H),5.25-5.39(s,1H),4.62–4.43(m,1H),4.20(s,1H),3.93–3.64(m,8 H),3.56(m,1H),3.50–3.30(m,3H),3.23(m,1H),3.13–2.94(m,2H),2.53(d,J=7.2Hz,3H),2.49–2.39 (m,4H),2.37–2.17(m,3H),2.16–1.97(m,3H),1.89(m,2H),1.72–1.17(m,10H),0.96(t,J=7.2Hz,6H).

[0278] (9) Synthesis of compound L-8-CPG

[0279]

[0280] At room temperature, HATU (60 mg, 0.156 mmol), DIEA (60 μL), lcaa-CPG (1000°A, 750 mg) were added to a solution of compound L-8 (150 mg, 0.152 mmol) in ACN (9.0 mL), and the mixture was shaken for 12 hours. After the reaction was completed, CPG was washed with ACN, and CAP A (acetic anhydride: tetrahydrofuran = 1:9, v / v, 3.0 mL) and CAP B (n-methylimidazole: pyridine: acetonitrile = 15:10:75, v / v / v, 3.0 mL) were added. The mixture was shaken at room temperature for 1 hour. The mixture was then filtered and washed 3 times with ACN (2 mL). The mixture was freeze-dried to obtain compound L-8-CPG (white powder, 750 mg).

[0281] (10) Synthesis of compound AS1411-ARL

[0282]

[0283] Compound L-8-CPG was placed in a synthesis column (60 mg*8) and synthesized by a KA H-8 solid phase synthesizer. The solid phase synthesis includes four steps: detritylation, coupling, capping and oxidation. After the reaction was completed, 1.5 mL of ammonia water was added to the CPG of each synthesis column and heated in an oven at 65°C for 16 hours. The supernatant was then collected and washed with water (1 mL*3). The crude product was purified by a protein purification system (Sepure, SDA) (column: Mono Q, 1.0 mL, method: mobile phase A: 40 mM NaOH aqueous solution, mobile phase B: 40 mM NaOH + 2.0 M NaCl aqueous solution) to obtain the target product AS1411-ARL (white powder, 9.93 mg, purity = 99.06%).

[0284] UPLC-MS(WATERS ACQUITY PREMIER):AS1411-ARL-UPLC,m / z=8918.84532[M] - (deconvolution); t R =10.537min(260nm).Mass error<50ppm.

[0285] HPLC:AS1411-ARL-HPLC,t R =13.487min(260nm), purity:99.061%.

[0286] Example 5: AS1411-ARL can degrade AR

[0287] (1) Human prostate cancer cells 22Rv1 were incubated with 0, 50, 100, 200, 500, and 1000 nM AS1411-ARL (prepared in Example 4). After 12 hours, the cell samples were collected, and the total protein was extracted using RIPA lysis buffer (purchased from: Biotech, catalog number: P0013B). The degradation of AR protein was detected by immunoblotting. The process is as follows: The protein sample was separated by SDS-PAGE electrophoresis, and the separated protein was transferred to a PVDF membrane. After blocking with TBST buffer containing 5% skim milk at room temperature for 1 hour, the primary antibody for AR (purchased from Millipore, catalog number 06-680) was incubated overnight at 4°C, and then washed with TBST. The membrane was incubated with HRP-labeled secondary antibody (purchased from: Abotek, catalog number: AS014) at room temperature for 1 hour, and the protein band blot was visualized using an enhanced chemiluminescence detection kit (purchased from: Abotek, catalog number: RM00021P). The results are as follows. Fig. 9 As shown in A, AS1411-ARL can reduce the protein level of AR in a concentration-dependent manner.

[0288] (2) Human prostate cancer cells 22Rv1 were incubated with 200 nM AS1411-ARL, and the corresponding cell samples were collected at 0, 1, 3, 6, 12, and 24 hours for protein extraction and subsequent immunoblotting experiments to detect AR protein degradation. Fig. 9 As shown in B, AS1411-ARL can reduce the protein level of AR in a time-dependent manner.

[0289] (3) Human prostate cancer cells 22Rv1 were incubated with solvent PBS or 200 nM AS1411, ARL, AS1411+ARL or AS1411-ARL. After 6 hours, cell samples were collected for protein extraction and subsequent immunoblotting to detect AR protein degradation. The results are shown in Fig. 9 As shown in C, only AS1411-ARL can reduce the protein level of AR.

[0290] Example 6: AS1411-ARL mediates AR degradation via the ubiquitin-proteasome pathway

[0291] Human prostate cancer cells 22Rv1 were incubated with solvent PBS or 200nM AS1411-ARL for 6 hours, and 10μM proteasome inhibitor MG132 was added. The corresponding cell samples were collected for protein extraction and subsequent immunoblotting experiments to detect protein degradation. The results are shown in Fig.10 As shown, MG132 can block the degradation of AR by AS1411-ARL, indicating that the degradation of AR by AS1411-ARL depends on the ubiquitin-proteasome pathway.

[0292] Example 7: AS1411-ARL relies on MDM2 and NCL to mediate AR degradation

[0293] (1) Negative control siRNA (siNC, SEQ ID NO: 5) or NCL siRNA (siNCL, SEQ ID NO: 6) was transfected into human prostate cancer cells 22Rv1 for 48 hours, and then the cells were incubated with solvent PBS or 200nM AS1411-ARL for 6 hours. The corresponding cell samples were collected for protein extraction and subsequent immunoblotting experiments to detect protein degradation. The results are shown in Fig.11 As shown in A, silencing NCL can reduce the degradation of AR by AS1411-ARL, indicating that AS1411-ARL relies on NCL to mediate the degradation of AR.

[0294] (2) Negative control siRNA (siNC, SEQ ID NO: 5) or MDM2 siRNA (siMDM2, SEQ ID NO: 7) were transfected into human prostate cancer cells 22Rv1, and the cells were then incubated with solvent PBS or 200 nM AS1411-ARL for 6 hours. The corresponding cell samples were collected for protein extraction and subsequent immunoblotting experiments to detect protein degradation. The results are shown in Figure 2. Fig.11 As shown in B, silencing MDM2 can reduce the degradation of AR by AS1411-ARL, indicating that AS1411-ARL relies on MDM2 to mediate the degradation of AR.

[0295] Example 8: AS1411-ARL inhibits tumor cell proliferation and promotes apoptosis

[0296] (1) Human prostate cancer cells 22Rv1 were inoculated into 96-well cell culture plates. During the experiment, the culture medium containing solvent PBS or 200 nM AS1411, ARL, AS1411+ARL or AS1411-ARL was replaced every day. The cell proliferation experiment was performed using a CCK-8 kit (purchased from MCE, catalog number HY-K0301). The absorbance at 450 nm was measured using an ELISA reader on the first, second, third and fourth days. The results are shown in Table 1. Fig.12 As shown in A, AS1411-ARL can inhibit the proliferation of human prostate cancer cell 22Rv1.

[0297] (2) Human prostate cancer cells 22Rv1 were inoculated into six-well plates for a cell apoptosis experiment. During the experiment, the culture medium containing solvent PBS or 200 nM AS1411, ARL, AS1411+ARL or AS1411-ARL was replaced every day. After 5 days, the cell apoptosis analysis was performed using the Annexin V-FITC and PI double staining cell apoptosis detection kit (purchased from Beyotime, catalog number C1062L). The results are shown in Figure 2. Fig.12As shown in B, AS1411-ARL can promote apoptosis of human prostate cancer cell 22Rv1.

[0298] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

[0299] The aforementioned embodiments and methods described in the present invention may be varied based on the ability, experience and preference of those skilled in the art.

[0300] In the present invention, merely listing the steps of the method in a certain order does not constitute any limitation on the order of the method steps.

Claims

1. An AR degrading agent or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate or deuterated compound thereof, wherein the AR degrading agent has the following structure: in, GRO is a guanine-rich oligonucleotide residue that can specifically bind to nucleolin; L is a linker; p is an integer from 1 to 100; Y 1 , Y 2 , Y 3 Independently selected from: single bond, -O-, -S-, -N(C0-C6 alkyl), -C(O)-, -C(S)-, -C(O)-(C0-C6 alkylene)-, -C(S)-(C0-C6 alkylene)-, -C(O)-N(C0-C6 alkyl)-, -C(S)-N(C0-C6 alkyl)-, SO, SO2; Ring A is an aromatic ring or an aromatic heterocyclic ring; Ring B is an aliphatic ring or a heterocyclic ring; Ring C is an aromatic ring or an aromatic heterocyclic ring; R1 is one or more independent substituents on the A ring, which are selected from: H, C1-C 10 Alkyl, -(C0-C6 alkylene)-(C3-C 10 Cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic group), halogen, cyano, nitro, azido, C1-C 10 Halogenated alkyl, C1-C 10 Haloalkoxy, -N(C 0-10 Alkyl)(C 0-10 Alkyl), -N(C 0-10 alkyl)CO(C 0-10 Alkyl), -N(C 0-10 alkyl)CON(C 0-10 Alkyl), -N(C 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C 0-10 Alkyl), -S(C 0-10 Alkyl), -SO(C 0-10 Alkyl), -SO2(C 0-10 Alkyl), -SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C 0-10 Alkyl)(C 0-10 alkyl), -CO(C 0-10 alkyl); R2 is one or more independent substituents on the B ring, which are selected from: H, C1-C 10 Alkyl, -(C0-C6 alkylene)-(C3-C 10 Cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic group), halogen, cyano, nitro, azido, C1-C 10 Halogenated alkyl, C1-C 10 Haloalkoxy, -N(C 0-10 Alkyl)(C 0-10 Alkyl), -N(C 0-10 alkyl)CO(C 0-10 Alkyl), -N(C 0-10 alkyl)CON(C 0-10 Alkyl), -N(C 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C 0-10 Alkyl), -S(C 0-10 Alkyl), -SO(C 0-10 Alkyl), -SO2(C 0-10 Alkyl), -SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C 0-10 Alkyl)(C 0-10 alkyl), -CO(C 0-10 alkyl); R3 is one or more independent substituents on the C ring, which are selected from: H, C1-C 10 Alkyl, -(C0-C6 alkylene)-(C3-C 10 Cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic group), halogen, cyano, nitro, azido, C1-C 10 Halogenated alkyl, C1-C 10 Haloalkoxy, -N(C 0-10 Alkyl)(C 0-10 Alkyl), -N(C 0-10 alkyl)CO(C 0-10 Alkyl), -N(C 0-10 alkyl)CON(C 0-10 Alkyl), -N(C 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C 0-10 Alkyl), -S(C 0-10 Alkyl), -SO(C 0-10 Alkyl), -SO2(C 0-10 Alkyl), -SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C 0-10 Alkyl)(C 0-10 alkyl), -CO(C 0-10 alkyl); Preferably, the GROs have a stable G4 structural feature.

2. The AR degradation agent according to claim 1, characterized in that The GROs is one of the DNA aptamer AS1411 and AS1411 derivatives / analogs; preferably, the AS1411 derivatives / analogs is one of GRO29A, GRO15A, AT11, GRO29-2, GRO29-3, GRO29-5, GRO29-13, GRO14C, GRO25A, GRO28A, GRO28B, GRO29-6, AT11-L0, AT21, AT27, GRO32A, GRO32B, GRO56A, GRO A, GRO B, GRO C, GRO D, GRO E, GRO F, GRO G, GRO H, GROI, GRO J, GRO K, GRO L, GRO M; preferably, the GROs is selected from: AS1411, GRO29A, GRO15A, AT11; Preferably, the GROs include chemical modifications, nucleic acid unit replacements or attachment of functional groups on the GROs; The chemical modification is that at least one base is modified, and the chemical modification includes at least one of phosphorylation, methylation, amination, sulfhydrylation, isotopization, thiophosphate backbone modification, methoxy modification, and fluorination modification; The nucleic acid unit is replaced by at least one nucleic acid unit being replaced by LNA, UNA or GNA; The functional group includes at least one of a fluorescent group, a radioactive group, a therapeutic drug, biotin, digoxin, a nanoluminescent material, a nucleic acid substance or an enzyme marker.

3. The AR degradation agent according to claim 1, characterized in that The GROs is AS1411.

4. The AR degradation agent according to any one of claims 1 to 3, characterized in that L has the following structure: in, L1 is a divalent group connected to GRO, which is selected from: a single bond, -O-(C0-C6 alkylene)-, -S-(C0-C6 alkylene)-, -N(R L1 )-(C0-C6 alkylene)-, -N(R L2 )C(O)-(C0-C6 alkylene)-, -OP(O)(OR L1 )O-(C0-C6 alkylene)-, -C(O)-(C0-C6 alkylene)-, -C(S)-(C0-C6 alkylene)-, -CON(R L1 )-(C0-C6 alkylene)-; L3 is a divalent group connected to the A ring, which is selected from: a single bond, -O-(C0-C6 alkylene)-, -S-(C0-C6 alkylene)-, -C(O)-(C0-C6 alkylene)-, -C(S)-(C0-C6 alkylene)-, -N(R L3 )-(C0-C6 alkylene)-, -CON(R L3 )-(C0-C6 alkylene)-, -N(R L3 )CO-(C0-C6 alkylene)-, -SO2-(C0-C6 alkylene)-, -SO-(C0-C6 alkylene)-, 4-10 membered heterocyclylene; L2 is a single bond or a divalent saturated or unsaturated straight or branched C1-C50 hydrocarbon chain, in which 0-6 methylene units are independently substituted by: -CY-, -O-, -S-, -SS-, -C(O)-, -C(S)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(O)N(R L2 )-、-N(R L2 )C(O)-、-N(R L2 )C(O)O-、-N(R L2 )C(O)N(R L2 )-、-N(R L2 )-、-S(O)2-、-S(O)2N(R L2 )-、-N(R L2 )S(O)2-、-S(O)-、-S(O)N(R L2 )-、-N(R L2 )S(O)-、-P(O)(OR L2 )O-、-P(O)-、-P(O)N(R L2 )-、-P(O)(N(R L2 )2)-、-OP(O)(OR L2 )2N(R L2 )-、-P(O)(OR L2 )2N(R L2 )-、-N(R L2 )P(O)(OR L2 )O-、-N(R L2 )P(O)-、-Si(R L2 )2-, -C(=N-CN)-, Amino acid residues, nucleotide residues, oligonucleotide residues, oligopeptide residues, wherein m2 is selected from an integer between 1 and 10, each -CY- is independently an optionally substituted divalent ring selected from the following: arylene, cycloalkylene, heterocyclylene; H in the hydrocarbon chain may be optionally substituted by one or more groups selected from the following: halogen, cyano, nitro, azido, -OR L0 、-C(O)R L0 、-C(S)R L0 、-C(O)OR L0 、-C(S)SR L0 、-OC(O)R L0 、-OC(S)R L0 、-OC(S)SR L0 、-C(O)N(R L0 )2、-OC(O)N(R L0 )2、-N(R L0 )C(O)OR L0 、-N(R L0 )SO2R L0 、-SO2N(R L0 )2、-OSO2N(R L0 )2、-N(R L0 )C(O)R L0 、-N(R L0 )2. -SR L0 、-SOR L0 、-SO2R L0 、-OSO2R L0 、C1-C 10 Alkyl, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C1-C 10 Halogenated alkyl, C1-C 10 Haloalkoxy, -(C0-C6 alkylene)-(C3-C 10 Cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclyl); R L0 , R L1 , R L2 and R L3 Independently selected from: H, C1-C 10 Alkyl, C2-C 10 Alkenyl, C2-C 10 Alkynyl, -(C0-C6 alkylene)-(C3-C 10 Cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic group), wherein the C1-C 10 Alkyl, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C0-C6 alkylene, C3-C 10 Cycloalkyl, C6-C 10 The H in the aryl group and the 4-10 membered heterocyclic group may be optionally substituted by one or more groups selected from the following: halogen, cyano, nitro, azido, hydroxyl, amino, thiol, carboxyl, C1-C 10 Alkyl, C2-C 10 Alkenyl, C2-C 10 Alkynyl, C1-C 10 Halogenated alkyl, C1-C 10 Haloalkoxy, -(C0-C6 alkylene)-(C3-C 10 Cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclyl); Preferably, L is linked to the 3' end, 5' end or any base, sugar or phosphate backbone in the middle of GRO.

5. The AR degradation agent according to any one of claims 1 to 3, characterized in that L2 is a C1-C20 straight chain alkylene group, wherein 0-6 methylene units in the alkylene group are independently substituted by the following groups: -O-, -S-, -SS-, -C(O)-, -C(O)O-, -OC(O)-, -N(R L2 )-、-C(O)N(R L2 )-、-N(R L2 )C(O)-、 Among them, each R L2 Independently selected from: H, C1-C6 alkyl, each R L4 Independently selected from: H, OH, C1-C6 alkoxy; Preferably, L2 can be selected from: C1-C20 straight chain alkylene, -(CH2CH2O) m2 -CH2-, -(CH2CH2O) m2 -CH2CH2-, -CH2-(CH2CH2O) m2 -CH2-, -CH2CH2-(CH2CH2O) m2 -CH2-, -CH2CH2-(CH2CH2O) m2 -CH2CH2-、-(C1-C 10 Alkylene)-O-(C1-C 10 Alkylene)-, -(C1-C 10 Alkylene)-NH-(C1-C 10 Alkylene)-, -(C1-C 10 Alkylene)-C(O)NH-(C1-C 10 Alkylene)-, -(C1-C 10 Alkylene)-NHC(O)-(C1-C 10 -(C1-C6 alkylene)-, -(C1-C6 alkylene)-O-(C1-C6 alkylene)-C(O)NH-(C1-C6 alkylene)-, -(C1-C6 alkylene)-O-(C1-C6 alkylene)-NHC(O)-(C1-C6 alkylene)-, wherein m2 is selected from an integer between 1 and 10, g is 0 or 1, h is selected from an integer between 0 and 10, i is selected from an integer between 0 and 10, and G is any suitable trivalent group; More preferably, L2 is selected from:

6. The AR degradation agent according to any one of claims 1 to 3, characterized in that L2 is selected from C1-C20 straight chain alkylene, -(C0-C6 alkylene)-(CH2CH2O) m2 -(C1-C6 alkylene)-, -(C1-C 10 Alkylene)-O-(C1-C 10 Alkylene)-, -(C1-C 10 Alkylene)-NH-(C1-C 10 Alkylene)-, -(C1-C 10 Alkylene)-C(O)NH-(C1-C 10 Alkylene)-, -(C1-C 10 Alkylene)-NHC(O)-(C1-C 10 -(C1-C6 alkylene)-, -(C1-C6 alkylene)-O-(C1-C6 alkylene)-C(O)NH-(C1-C6 alkylene)-, -(C1-C6 alkylene)-O-(C1-C6 alkylene)-NHC(O)-(C1-C6 alkylene)-, wherein one methylene unit in the alkylene is independently Substitution, G is any suitable trivalent group; Preferably, L2 is The x end is connected to L3, the y end is connected to L1, the J ring is a 4-6 membered saturated heterocyclic ring, m2 is selected from an integer between 1 and 10, h is selected from an integer between 0 and 10, and i is selected from an integer between 0 and 10; More preferably, L2 is More preferably, L2 is selected from:

7. The AR degradation agent according to any one of claims 1 to 3, characterized in that L2 is a C1-C20 straight chain alkylene group, wherein 1-3 methylene units are independently substituted by the following groups: -CY-, Optionally, L2 further comprises a group selected from the following: -O-, -C(O)-, -N(R L2 )-、-C(O)N(R L2 )-、-N(R L2 )C(O)-、 Among them, each R L2 Independently selected from: H, C1-C6 alkyl, each R L4 Independently selected from: H, OH, C1-C6 alkoxy; Preferably, the -CY- is selected from: More preferably, L2 is selected from:

8. The AR degradation agent according to any one of claims 4 to 7, characterized in that: L1 is a single bond or O.

9. The AR degradation agent according to any one of claims 4 to 7, characterized in that: L3 is selected from: a single bond, -N(H)-, 10. The AR degradation agent according to any one of claims 4 to 8, characterized in that: Part of R2', R2", R2'', R3' are independently selected from: H, C1-C 10 Alkyl, -(C0-C6 alkylene)-(C3-C 10 Cycloalkyl), -(C0-C6 alkylene)-(C6-C 10 aryl), -(C0-C6 alkylene)-(4-10 membered heterocyclic group), halogen, cyano, nitro, azido, C1-C 10 Halogenated alkyl, C1-C 10 Haloalkoxy, -N(C 0-10 Alkyl)(C 0-10 Alkyl), -N(C 0-10 alkyl)CO(C 0-10 Alkyl), -N(C 0-10 alkyl)CON(C 0-10 Alkyl), -N(C 0-10 Alkyl)SO2(C 0-10 Alkyl), -O(C 0-10 Alkyl), -S(C 0-10 Alkyl), -SO(C 0-10 Alkyl), -SO2(C 0-10 Alkyl), -SO2N(C 0-10 Alkyl)(C 0-10 Alkyl), -COO(C 0-10 Alkyl), -OCO(C 0-10 Alkyl), -CON(C 0-10 Alkyl)(C 0-10 alkyl), -CO(C 0-10 alkyl).

11. The AR degradation agent according to claim 10, characterized in that R1 is selected from: H, C1-C6 alkyl, halogen, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, preferably H; R2 is selected from: H, C1-C6 alkyl, halogen, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, preferably H; R3 is selected from: H, cyano, C1-C6 alkyl, halogen, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, preferably halogen or cyano; R2' is selected from: H, C1-C6 alkyl, halogen, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, preferably H; R2" is selected from: H, C1-C6 alkyl, halogen, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, preferably H; R2"' is selected from: H, C1-C6 alkyl, halogen, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, preferably H; R3' is selected from the group consisting of: H, cyano, C1-C6 alkyl, halogen, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, preferably halogen or cyano.

12. The AR degradation agent according to any one of claims 4 to 8 and 11, characterized in that: The AR degradation agent has the following structure: Preferably, the AR degradation agent has the following structure:

13. The AR degradation agent according to any one of claims 1 to 3, characterized in that: The AR degradation agent has the following structure: Wherein, h is selected from an integer between 0 and 10, and i is selected from an integer between 0 and 10; Preferably, the AR degrading agent is selected from the following structures:

14. A pharmaceutical composition comprising the AR degrading agent according to any one of claims 1 to 13 or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate or deuterated compound thereof, and one or more pharmaceutically acceptable excipients.

15. A delivery system for an AR degrading agent, comprising the AR degrading agent according to any one of claims 1 to 13 and a carrier; Preferably, the vector is a viral vector or a non-viral vector; More preferably, the viral vector is a lentivirus, adenovirus or adeno-associated virus vector; More preferably, the non-viral vector is a lipid nanoparticle (LNP), a polymer nanocarrier, an inorganic nanocarrier, a protein carrier or an exosome.

16. Use of the AR degrading agent according to any one of claims 1 to 13 or a pharmaceutically acceptable salt, stereoisomer, ester, prodrug, solvate or deuterated compound thereof in the preparation of a medicament for preventing and / or treating AR-related diseases; Preferably, the disease is selected from the group consisting of: tumors, autoimmune diseases, inflammatory diseases, diseases associated with pathogen infection, neurodegenerative diseases, cardiovascular diseases, metabolic diseases, fibrotic diseases, and neuro-psychiatric disorders; More preferably, the tumor is selected from the group consisting of lung cancer, malignant melanoma, brain tumor, tumor of digestive organs, uterine cancer, endometrial cancer, testicular cancer, palate cancer, pharyngeal cancer, tongue cancer, oral cancer, various sarcomas, osteosarcoma, blood system tumors, nervous system tumors, brain glioma, glioblastoma, skin cancer, skin appendage cancer and skin metastasis, medulloblastoma, blastoma, liposarcoma, neuroendocrine tumor, synovial cell sarcoma, gastrinoma, carcinoid tumor, mesothelioma, islet cell carcinoma, Schwannoma, meningioma, melanoma, acoustic neuroma, adenocarcinoma, lymphoid malignancies, epithelial squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer, squamous cell carcinoma, lung adenocarcinoma, peritoneal cancer, lung squamous cell carcinoma, hepatocellular carcinoma, stomach cancer, intestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, thyroid cancer, bladder cancer, breast cancer, metastatic breast cancer, colon cancer, rectal cancer, prostate cancer, salivary gland cancer, kidney cancer, vulvar cancer, anal cancer, penile cancer, esophageal cancer, biliary tract tumors, and head and neck cancer; More preferably, the autoimmune disease is selected from the group consisting of organ-specific autoimmune disease, systemic lupus erythematosus, rheumatoid arthritis, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease, autoimmune hemolytic anemia, thyroid autoimmune disease, and ulcerative colitis; More preferably, the inflammatory disease is selected from the group consisting of osteoarthritis, acute gout, multiple sclerosis, inflammatory bowel disease (e.g., Crohn's disease and ulcerative colitis), neuroinflammation, asthma, chronic obstructive airway disease, pneumonia, myositis, eczema, dermatitis, acne, cellulitis, occlusive disease, thrombosis, alopecia, nephritis, vasculitis, retinitis, uveitis, scleritis, sclerosing cholangitis, hypophysitis, thyroiditis, septic shock, systemic inflammatory response syndrome (SIRS), toxic shock syndrome, acute lung injury, ARDS (adult respiratory distress syndrome), acute renal failure, burns, pancreatitis (e.g., acute pancreatitis), postoperative syndrome, sarcoidosis, Herxheimer reaction, encephalitis, myelitis, meningitis, and malaria; More preferably, the neurodegenerative disease is selected from the group consisting of: Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), different types of spinocerebellar ataxia (SCA), Pick's disease; More preferably, the pathogen infection-related disease is selected from: influenza, SARS, COVID-19, viral hepatitis (such as hepatitis A, hepatitis B, hepatitis C, hepatitis D, etc.), AIDS, rabies, dengue fever, Ebola virus disease; More preferably, the cardiovascular disease is selected from the group consisting of coronary heart disease, peripheral arterial disease, atherosclerosis, ischemic heart disease, ischemic cardiomyopathy, myocardial infarction, heart failure, angina pectoris, myocarditis, hypercholesterolemia, hypertension, ischemia-reperfusion injury, cerebrovascular ischemia (stroke), embolism (e.g., pulmonary embolism, renal embolism, hepatic embolism, gastrointestinal embolism or peripheral limb embolism) or myocardial ischemia; More preferably, the metabolic disease is selected from the group consisting of: diabetes (such as type I diabetes, type II diabetes or gestational diabetes), obesity, insulin resistance, hyperinsulinemia, fatty liver (NASH or other), cachexia, hypercholesterolemia, gout; More preferably, the fibrotic disease is selected from the group consisting of: myocardial fibrosis, pulmonary fibrosis, renal fibrosis, postoperative stenosis, keloid formation, cirrhosis, biliary cirrhosis, scleroderma; More preferably, the neuro-psychiatric disorder is selected from: anxiety, depression; More preferably, the diseases further include polycystic ovary syndrome, hirsutism, acne, androgenic alopecia, infertility, pregnancy loss, gestational diabetes, gestational hypertension, hyperandrogenism, pelvic floor dysfunction, and pelvic pain syndrome.

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