Application of a cdc20 small molecule ligand compound and its degrader

Abstract

The application belongs to the technical field of pharmaceutical chemistry, and particularly relates to a compound capable of combining with cell division cycle 20 (Cdc20) and a composition thereof, and particularly discloses a compound capable of combining with cell division cycle 20 (Cdc20), wherein the compound is shown as (I); the compound can further become a targeted protein degradation agent and a pharmaceutical composition, and is used for treating or preventing diseases caused by promoting abnormal cell division after Cdc20 activates an APC complex, and the compound has high Cdc20 combining ability.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical chemistry technology, specifically relating to a compound and its composition that can bind to cell division cycle protein 20 (Cdc20), and also to a protein degrading agent based on a compound that can bind to cell division cycle protein 20 (Cdc20) as an E3 ubiquitin ligand ligand and its application. Background Technology

[0002] Cancer has become one of the leading causes of death worldwide, seriously threatening human health. Currently, commonly used cancer treatments include surgery, chemotherapy, radiotherapy, and targeted therapy, all of which have made significant progress, greatly improving the survival of cancer patients. With the development of molecular biology and related technologies, scientists have gained a deeper understanding of the occurrence, development, and related targets of tumors. The view that "malignant tumors are a type of cell cycle disease" has also gained widespread acceptance. Therefore, anti-tumor drugs targeting cell cycle regulation, designed based on this view, have become a research hotspot in the field of oncology.

[0003] The cell cycle is a fundamental process of cellular life, divided into: G1 phase (pre-DNA synthesis), S phase (DNA synthesis phase), G2 phase (post-DNA synthesis phase), and M phase (mitosis phase). Precise regulation of the cell cycle is essential for ensuring genomic stability and normal cellular function, involving a complex information network system. Cell cycle regulation mechanisms are closely related to tumorigenesis; many cell cycle regulatory proteins are altered and inactivated during tumorigenesis, leading to genomic instability, typically manifested as gene mutations, gene deletions, gene rearrangements and translocations, as well as centrosome amplification and chromosomal aberrations. Genomic instability results in uncontrolled cell proliferation and tumorigenesis. Based on this theory, many drugs have been developed to inhibit different stages of the cell cycle for tumor treatment.

[0004] During cell division, cells continuously produce new proteins and degrade old ones. Protein degradation typically occurs via the ubiquitin-proteasome pathway. This pathway involves three enzymes: ubiquitin-activating enzyme (E1), ubiquitin-conjugating enzyme (E2), and ubiquitin-ligating enzyme (E3). Through this reaction, the protein binds to a polyubiquitin chain and is then degraded via the 26S proteasome. The SCF complex (Skp1 / Cullin / F-box protein) and the Anaphase promoting complex / cyclosome (APC / C) are two E3 ubiquitin ligases that play key roles in cell cycle regulation. APC / C is the largest E3 ubiquitin ligase, composed of at least 14 subunits and one co-activator. Upon activation by cyclin 20 (Cdc20) or Cdc20 homologue 1 (Cdh1), it forms two distinct E3 ubiquitin ligase complexes, APC. Cdc20 or APC Cdh1 APC Cdc20 APC / C regulates cell cycle progression by disrupting key cell cycle regulators during the transition from metaphase to anaphase, with APCCdh1 playing a crucial role in the M and G1 phases. APC / C participates in the degradation of various proteins, including cyclins and cyclin-dependent kinases, via the ubiquitin-proteasome system, promoting cell cycle progression. Cdc20 is a positive regulator of APC / C, directly binding to substrates and presenting them to APC / C, thereby enabling substrate degradation via the ubiquitin-proteasome pathway.

[0005] Furthermore, Cdc20 is also a crucial component of the spindle assembly checkpoint (SAC). As an important molecular monitoring mechanism within the cell cycle checkpoints, the SAC is complex, primarily composed of serine / threonine protein kinases (budding uninhibited by benzimidazoles-1, BUB1), mitotic arrest deficient-2 (MAD2), and Cdc20. During mitosis, the SAC precisely regulates the dynamic balance between kinetic chordates and microtubules, ensuring that after chromosome replication, genetic material is stably and accurately distributed to each daughter cell. The SAC effectively prolongs mitosis, promoting correct chromosome attachment to the spindle microtubules. Cdc20, as a key gene regulating cell cycle progression, can activate the cell cycle APC / C during mitosis, thereby inducing chromosome separation and entering the later stages of mitosis. When chromosome segregation is abnormal, SAC is activated. The MAD2-MAD1 complex in the cytoplasm interacts with free O-MAD2, transforming into C-MAD2, which then binds to Cdc20. This C-MAD2 then forms the HCC complex with BubR1 / MAD3-BUB3, which effectively inhibits APC. Cdc20 The activity of the chromosomes prevents them from separating further. Only after all chromosomes are properly aligned can the cell continue to divide and form daughter cells.

[0006] The human Cdc20 gene (cell division cycle 20homolog, also known as p55CDC) is located in the human chromosome 1p34.1 region, and its genome sequence contains 11 exons and 10 introns. The human Cdc20 protein consists of 499 amino acid residues and contains 7 repeat sequences of WD40. Studies have shown that Cdc20 is undetectable in the G1 phase of mitosis, begins to appear and accumulate in late S phase, reaches its peak in G2 / M phase, and remains active until late mitosis. Cdh1 It ubiquitinates and degrades Cdc20. In recent years, as the functions of Cdc20 in ubiquitination and degradation of proteins and its interaction with spindle assembly checkpoints have been gradually revealed, its role in cell cycle and cell proliferation has become increasingly important. Therefore, research on Cdc20 has received more and more attention and in-depth study.

[0007] With the continuous development and progress of clinical molecular biology, the high expression of cyclin 20 (Cdc20) in various malignant tumors has been identified as an important biological factor promoting tumorigenesis and development. Studies have reported that Cdc20 expression in various tumor tissues, including breast cancer, colon cancer, gastric cancer, non-small cell lung cancer, rectal cancer, and liver cancer, is significantly higher than in normal tissues; moreover, its expression level is closely related to the pathological severity and prognosis of tumor patients, with higher Cdc20 expression levels associated with greater risk. Currently, many substrates of Cdc20 have been reported, some of which are proteins directly involved in regulating cell cycle processes, such as securin, cyclin A, cyclin B, and transcription factor E2F1. Cdc20 Ubiquitination of cell cycle proteins such as securin and cyclin B degrades them, disrupting normal cell division. Therefore, designing small molecule inhibitors targeting Cdc20 to disrupt the interaction between Cdc20 and its substrate may achieve anti-tumor effects.

[0008] The development of novel target protein chimeric degradation molecules (PROTACs) is a field of great interest in current drug development. PROTACs consist of three parts: a target protein ligand, an E3 ubiquitin ligase ligand, and a linker chain. They can recognize the target protein and the E3 ubiquitin ligase through the target protein ligand and the E3 ubiquitin ligase ligand, respectively, leading to polyubiquitination of the target protein, thereby enabling the target protein to be recognized and cloned by the proteasome. Currently, PROTACs mainly use small molecule E3 ubiquitin ligase ligands. These novel PROTACs can rapidly diffuse into cells, have higher affinity for the target protein, and stronger metabolic stability (Meng Yifan et al., Research progress of small molecule PROTACS that induce protein degradation; Guangdong Chemical Industry; 2018, 45(05), 125-126). PROTACs, as bifunctional molecules, consist of a small compound that can bind to the target protein (POI), with a linker group introduced at the appropriate position, and then linked to a small molecule compound that can bind to E3 ubiquitinase. The resulting small molecule probe can bind to both the target protein and E3 ubiquitinase simultaneously, thereby ubiquitinizing the protein. The multiubiquitinated protein can be recognized and degraded by the proteasome. More than 600 E3 ubiquitinases have been discovered, but currently, only a limited number such as MDM2, clAP-1, CRBN, and VHL are most commonly used in the design of PROTACs (Xu Pengfei et al., Research progress of small molecule PROTACs that induce protein degradation, Chinese Journal of New Drugs [J]. 2017, 26(22)). The key advantage of this protein degradation technology compared to small molecule ubiquitin ligase inhibitors is that PROTACS exhibits specific sensitivity to drug resistance targets. Unlike simple small molecule inhibitors, PROTACs affect enzyme activity and non-enzyme function by clearing target proteins (Zou Y, Ma D, Wang Y. The PROTAC technology in drug development[J]. Cell Biochem Funct, 2019, 37(1): 21-30). After entering the cell, the POI ligand in the structure of PROTAC molecules can specifically bind to the corresponding target protein, while the other end can recruit E3 ligase to form a POI-PROTAC-E3 ligase ternary complex, in which E3 ligase can mediate ubiquitination of POI by ubiquitin-binding enzyme E2. After the ternary complex dissociates, the ubiquitin-tagged POI is recognized and degraded by the proteasome, thereby selectively reducing the level of the target protein. Furthermore, PROTACs can be recycled multiple times in the cell and can be repeatedly used in vivo, achieving good therapeutic effects at low doses.

[0009] APC Cdc20The complex is the largest E3 ubiquitin ligase in vivo, and Cdc20 is highly expressed in various tumor tissues. Therefore, APC... Cdc20 It has the potential to be used as a novel E3 ubiquitin ligase for the development of protein degrading agents. Currently, only two small-molecule inhibitors of CDc20 have been reported: TAME and Apcin. TAME is a non-specific inhibitor; it works by disrupting the binding of Cdc20 to APC / C, and it can also disrupt the binding of Cdh1 to APC / C. Apcin is currently the only reported Cdc20-specific inhibitor, which works by specifically inhibiting the binding of the substrate to Cdc20. However, its binding affinity to Cdc20 is low (in the hundreds of micromoles), and its antitumor activity is very weak. Therefore, developing highly active, specific Cdc20 ligands for antitumor activity and for the development of protein degrading agents are of great significance. Summary of the Invention

[0010] In a first aspect, the present invention provides a compound that can bind to cell division cycle protein 20 (Cdc20), said compound as shown in (I);

[0011]

[0012] In structural formula I:

[0013] X1, X2, and X3 can be one or two nitrogen atoms and the rest carbon atoms, or all of them can be carbon atoms;

[0014] R1, R2, R3, R4, and R5 can be hydrogen atoms (-H), hydroxyl groups (-OH), methoxy groups (-OCH3), ethoxy groups (-OCH2CH3), propoxy groups (-OCH2CH2CH3), or cyclopropoxy groups. Halogen atoms (F, Cl, Br), monofluoromethyl (-CH2F), difluoromethyl (-CHF2), trifluoromethyl (-CF3), trifluoromethoxy (-OCF3), cyano (-CN), amino (-NH2), nitro (-NO2), C1-C6 straight-chain, branched, cycloalkyl or C1-C6 unsaturated straight-chain, branched, cycloalkenyl, alkynyl, etc.;

[0015] Y1, Y2, Y3, and Y4 can have one or two nitrogen atoms and the rest carbon atoms, or all of them can be carbon atoms;

[0016] R6, R7, R8, and R9 can be hydrogen atoms (-H), hydroxyl groups (-OH), methoxy groups (-OCH3), ethoxy groups (-OCH2CH3), propoxy groups (-OCH2CH2CH3), or cyclopropoxy groups. Halogen atoms (F, Cl, Br), monofluoromethyl (-CH2F), difluoromethyl (-CHF2), trifluoromethyl (-CF3), trifluoromethoxy (-OCF3), cyano (-CN), amino (-NH2), nitro (-NO2), C1-C6 straight-chain, branched, cycloalkyl or C1-C6 unsaturated straight-chain, branched, cycloalkenyl, alkynyl, etc.;

[0017] X4, X5, and X6 can be one or two nitrogen atoms and the rest carbon atoms, or all carbon atoms;

[0018] When Y5, Y6, Y7, and Y8 are all carbon atoms, its structure is: etc., corresponding substituent R 10 R 11 R 12 R 13 It can be simultaneously or partially composed of hydrogen atoms (-H), hydroxyl groups (-OH), methoxy groups (-OCH3), ethoxy groups (-OCH2CH3), propoxy groups (-OCH2CH2CH3), or cyclopropoxy groups. Halogen atoms (F, Cl, Br), monofluoromethyl (-CH2F), difluoromethyl (-CHF2), trifluoromethyl (-CF3), trifluoromethoxy (-OCF3), cyano (-CN), amino (-NH2), nitro (-NO2), C1-C6 straight-chain, branched, cycloalkyl or C1-C6 unsaturated straight-chain, branched, cycloalkenyl, alkynyl, etc.;

[0019] When any one of the substituents in Y5, Y6, Y7, and Y8 is a carbonyl group (-CO-), and the remaining substituents are atoms such as C, N, O, and S, or when none of the substituents in Y5, Y6, Y7, and Y8 are present;

[0020] When any one of the substituents in Y5, Y6, Y7, and Y8 is -CO-, O, S, or is absent, its corresponding substituent R 10 R 11 R 12 R 13 It does not exist;

[0021] When any one of the substituents in Y5, Y6, Y7, and Y8 is C or N, its corresponding substituent R 10 R 11 R 12 R 13 It can be simultaneously or partially composed of hydrogen atoms (-H), hydroxyl groups (-OH), methoxy groups (-OCH3), ethoxy groups (-OCH2CH3), propoxy groups (-OCH2CH2CH3), or cyclopropoxy groups. Halogen atoms (F, Cl, Br), monofluoromethyl (-CH2F), difluoromethyl (-CHF2), trifluoromethyl (-CF3), trifluoromethoxy (-OCF3), cyano (-CN), amino (-NH2), nitro (-NO2), C1-C6 straight-chain, branched, cycloalkyl, or C1-C6 unsaturated straight-chain, branched, cycloalkenyl, alkynyl, etc., representative structures are as follows: wait;

[0022] When Y5, Y6, Y7, and Y8 are all absent, the structure is: Corresponding substituent R 10 R 11 It can be simultaneously or partially composed of hydrogen atoms (-H), hydroxyl groups (-OH), methoxy groups (-OCH3), ethoxy groups (-OCH2CH3), propoxy groups (-OCH2CH2CH3), or cyclopropoxy groups. Halogen atoms (F, Cl, Br), monofluoromethyl (-CH2F), difluoromethyl (-CHF2), trifluoromethyl (-CF3), trifluoromethoxy (-OCF3), cyano (-CN), amino (-NH2), nitro (-NO2), C1-C6 straight-chain, branched, cycloalkyl or C1-C6 unsaturated straight-chain, branched, cycloalkenyl, alkynyl, etc.;

[0023] R 14 R 15 It can be a hydrogen atom (-H), a hydroxyl group (-OH), a methoxy group (-OCH3), an ethoxy group (-OCH2CH3), a propoxy group (-OCH2CH2CH3), or a cyclopropoxy group. Halogen atom (F, Cl, Br), monofluoromethyl (-CH2F), difluoromethyl (-CHF2), trifluoromethyl (-CF3), trifluoromethoxy (-OCF3), cyano (-CN), amino (-NH2), nitro (-NO2), C1-C6 straight-chain, branched, or cycloalkyl groups, C1-C6 unsaturated straight-chain, branched, cycloalkenyl, or alkynyl groups, substituted or unsubstituted furans Thiophene Pyrrole imidazole pyrazole Triazole Tetrazolium Oxazole Isoxaazole Thiazole Isothiazine benzene ring Pyridine Pyrimidine pyridazine Aromatic rings, substituted or unsubstituted tetrahydrofurans Tetrahydrothiophene Tetrahydropyrrole Tetrahydroimidazole Pyrazolidine Oxazolidine Isoxazolide Thiazolidine Isothiazolidine pyran Tetrahydropyran Piperidine Aromatic ring furan pyran Piperidine, Piperazine Five- or six-membered heterocycles;

[0024] W1 and W2 can be nitrogen-hydrogen (-NH-), oxygen (-O-)amide (-CONH-,-NHCO-), sulfonamide (-NHSO2-,-SO2NH-), sulfinamide (-NHSO-,-SONH-), etc.

[0025] n1 and n2 are carbon chains with lengths of 1 to 5, and the chiral carbon 1 has an R-type or S-type configuration.

[0026] Furthermore, the general formula compound may specifically have the following structure:

[0027]

[0028]

[0029] In a second aspect, the present invention also provides a pharmaceutically acceptable salt of the compound (I) of the first aspect of the present invention.

[0030] Furthermore, the pharmaceutically acceptable salt refers to a salt formed by the addition of a non-toxic acid or its base to the parent compound.

[0031] Thirdly, the present invention provides a protein degrading agent, which is composed of a target protein ligand + a linker + a Cdc20 protein conjugate (ligand); the expression of the protein degrading agent is shown in (II):

[0032]

[0033] Wherein, the Cdc20 protein conjugate (ligand) is the compound of formula (I) that can bind cell division cycle protein 20 (Cdc20) as described in the first aspect of the present invention;

[0034] The target protein ligand can be selected from the following:

[0035] Target 1: AR

[0036]

[0037] Target 2: ER

[0038]

[0039] Target 3: BRAF, BRAF V600E

[0040]

[0041] Target 4: E GFR

[0042]

[0043] Target 5: F GFR

[0044]

[0045] Target 6: eEF2K

[0046]

[0047] Target 7: eIF4E

[0048]

[0049] Target 8: Src / IGF-1R

[0050]

[0051] Target 9: KRAS G12C

[0052]

[0053] Target 10: KRAS G12D

[0054]

[0055] Target 11: SOS1

[0056]

[0057] Target 12: GRB2

[0058]

[0059] Target 13: MEK

[0060]

[0061] Target 14: Myc

[0062]

[0063] Target 15: p38

[0064]

[0065] Target 16: p300 / CBP

[0066]

[0067] Target 17: PDEδ

[0068]

[0069] Target 18: SHP2

[0070]

[0071] Target 19: AKT

[0072]

[0073] Target 20: ALK

[0074]

[0075] Target 21: Bcl-xl / Bcl-2

[0076]

[0077] Target 22: BCR-ABL

[0078]

[0079] Target 23: FAK

[0080]

[0081] Target 24: MDM2

[0082]

[0083] Target 25: FLT3

[0084]

[0085] Target 26: JAK

[0086]

[0087] Target 27: STAT3

[0088]

[0089] Target 28: STAT5

[0090]

[0091] Target 29: FOXM1

[0092]

[0093] Target 30: BRD

[0094]

[0095] Target 31: ENL

[0096]

[0097] Target 32: HDAC6

[0098]

[0099] Target 33: HDAC1 / 2 / 3

[0100]

[0101] Target 34: KDM5C

[0102]

[0103] Target 35: NAMPT

[0104]

[0105] Target 36: NSD3

[0106]

[0107] Target 37: PRC2 (EZH2, EED)

[0108]

[0109] Target 38: PRMT5

[0110]

[0111] Target 39: SIRT2

[0112]

[0113] Target 40: WDR5

[0114]

[0115] Target 41: Aurora A

[0116]

[0117] Target 42: Aurora B

[0118]

[0119] Target 43: CDK2

[0120]

[0121] Target 44: CDK2 / 5

[0122]

[0123] Target 45: CDK2 / 4 / 6

[0124]

[0125] Target 46: CDK9

[0126]

[0127] Target 47: CDK12

[0128]

[0129] Target 48: Wee1

[0130]

[0131] Target 49: hRpn13 Pru

[0132]

[0133] Target 50: ZFP91

[0134]

[0135] Target 51: BTK

[0136]

[0137] Target 52: CCR9

[0138]

[0139] Target 53: CD147

[0140]

[0141] Target 54: CRABP

[0142]

[0143] Target 55: HSP90

[0144]

[0145] Target 56: IDO1

[0146]

[0147] Target 57: LXR-β

[0148]

[0149] Target 58: MIF

[0150]

[0151] Target 59: PARP1

[0152]

[0153] Target 60: PARP14

[0154]

[0155] Target 61: PD-L1

[0156]

[0157] Target 62: PLK1

[0158]

[0159] Target 63: RAR

[0160]

[0161] Target 64: RIPK2

[0162]

[0163] Target 65: SF3B1

[0164]

[0165] Target 66: SLC

[0166]

[0167] Target 67: SMARCA2 / 4

[0168]

[0169] Target 68: Tyrosinase

[0170]

[0171] Target 69: SARS-CoV-2

[0172]

[0173] Target 70: HDAC1 / 2 / 3

[0174]

[0175] Target 71: H-PGDS

[0176]

[0177] Target 72: IRAK1

[0178]

[0179] Target 73: IRAK3

[0180]

[0181] Target 74: IRAK4

[0182]

[0183] Target 75: GSK-3β

[0184]

[0185] Target 76: LRRK2

[0186]

[0187] Target 77: α-Synuclein

[0188]

[0189] Target 78: Tau

[0190]

[0191] Target 79: TRKA

[0192]

[0193] Target 80: TRKC

[0194]

[0195] Target 81: HMGCR

[0196]

[0197] And / or target 82: VEGFR2

[0198]

[0199] The Linker sub-linker is selected from the following:

[0200] Linker 1 Linker 2 Linker 3 or

[0201] Linker 4

[0202] In Linker 1, A1 and B1 are selected from one or more of the following: -O-, -CO-, -NH-, -NH-CO-, -CO-NH-, -S- and / or none; m1 is selected from an integer from 0 to 20.

[0203] In Linker 2, A2 and B2 are selected from one or more of -O-, -CO-, -NH-, -NH-CO-, -CO-NH-, -S- and / or none; m2 is selected from an integer from 0 to 20.

[0204] In Linker 3, A3, A4 and B3, B3 are selected from one or more of -O-, -CO-, -NH-, -NH-CO-, -CO-NH-, -S- and / or none; m3 and m4 are selected from integers from 0 to 20.

[0205] In Linker 4, A5, A6 and B5, B6 are selected from one or more of -O-, -CO-, -NH-, -NH-CO-, -CO-NH-, -S- and / or none; m5 and m6 are selected from integers from 0 to 20.

[0206] In Linker 3 and Linker 4, C1 and C2 are selected from the following:

[0207]

[0208]

[0209]

[0210] Fourthly, the present invention provides a pharmaceutical composition comprising a combination of a compound of formula (I) of the first aspect or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable diluent or carrier.

[0211] Furthermore, the pharmaceutical composition may also comprise an enantiomer, diastereomer, stereoisomer, or pharmaceutically acceptable salt of a compound of formula (I) combined with a pharmaceutically acceptable diluent or carrier.

[0212] Fifthly, the present invention provides a pharmaceutical composition for targeting and degrading proteins, the pharmaceutical composition comprising a combination of a compound of formula (II) or a pharmaceutically acceptable salt thereof with a pharmaceutically acceptable diluent or carrier.

[0213] Furthermore, the pharmaceutical composition may also comprise an enantiomer, diastereomer, stereoisomer, or pharmaceutically acceptable salt of a compound of formula (II) combined with a pharmaceutically acceptable diluent or carrier.

[0214] In a sixth aspect, the present invention provides a treatment method for treating or preventing pathological conditions or symptoms in mammals (such as humans), such as a method for treating diseases caused by abnormal cell division after activation of the APC complex by Cdc20.

[0215] Furthermore, the diseases mentioned include, but are not limited to, mammalian tumors and neurodegenerative diseases.

[0216] Furthermore, the tumors include, but are not limited to, adrenocortical carcinoma, prostate cancer, intrahepatic bile duct cancer, bladder cancer, breast cancer, cervical cancer, colorectal cancer, lymphoma, Hodgkin's lymphoma, liver cancer, non-small cell lung cancer, small cell lung cancer, gastrointestinal cancer, head and neck cancer, neuroendocrine tumors, and kidney cancer.

[0217] Furthermore, the neurodegenerative diseases include, but are not limited to, Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), etc.

[0218] In a seventh aspect, the invention also includes a method for inhibiting abnormal cell division in tumors and neurodegenerative diseases, comprising administering an effective amount of at least one compound of formula I or formula II or a pharmaceutically acceptable salt thereof to the mammal in need of treatment for such diseases.

[0219] Eighthly, the present invention provides a method for adjusting the amount of an APC complex activated by Cdc20, the method comprising administering to a subject in need an effective amount of the compound (I), compound (II), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer of the present application.

[0220] Furthermore, the method includes administering an effective amount of the compound (I), compound (II), or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer of the present application to a subject in need.

[0221] In a ninth aspect, the present invention provides a medicament for treating or preventing diseases caused by abnormal cell division promoted after Cdc20-activated APC complexes are activated.

[0222] Furthermore, the drug comprises the compound (I), compound (II) of the present invention, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof.

[0223] In a tenth aspect, the present invention provides an inhibitor for the treatment or prevention of APC complexes activated by Cdc20.

[0224] Furthermore, the inhibitor comprises the compound (I), compound (II) of the present invention, or a pharmaceutically acceptable salt, hydrate, solvate, prodrug, stereoisomer, or tautomer thereof. Attached Figure Description

[0225] Figure 1 The results of the Cdc20 ligand affinity test are for Example 10.

[0226] Figure 2 Example 11: Cdc20 ligand inhibits tumor cell proliferation.

[0227] Figure 3 Low electrophoretic patterns of protein degradation at the same concentration of different compounds in human prostate cancer cell line LNCaP.

[0228] Figure 4 Low electrophoretic maps of protein degradation of different compounds at different concentrations in human prostate cancer cell line LNCaP.

[0229] Figure 5 To detect the inhibitory effect of MJC636 on the proliferation of prostate cancer cells.

[0230] Figure 6 The effect of bromine domain protein (BRD) on human chronic myeloid monocytic leukemia MV-4-11 cells.

[0231] Figure 7 The degradation of BRD4 protein by different compounds.

[0232] Figure 8 To detect the inhibitory effect of MJC108 on cancer cell proliferation. Detailed Implementation

[0233] The present invention will be described below through specific embodiments, but the present invention is not limited thereto. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

[0234] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0235] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0236] The following lists the definitions of various terms used in this application. These definitions apply to the terminology used throughout this specification and claims, unless otherwise limited individually or as part of a larger group in particular cases.

[0237] The term “stereoisomer” as used in this article refers to compounds that have the same chemical composition but different spatial arrangements of atoms or groups, including “diastereomers” and “enantiomers”.

[0238] The term "diastereomer" as used herein refers to a stereoisomer having two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers possess different physical properties, such as melting point, boiling point, spectral characteristics, and reactivity. In the presence of resolving agents or chromatography, mixtures of diastereomers can be separated using high-resolution analytical steps such as electrophoresis and crystallization, in the presence of chiral HPLC columns.

[0239] The term "enantiomer" as used herein refers to two stereoisomers of a compound that do not overlap as mirror images of each other. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can occur during chemical reactions or processes where stereoselectivity or stereoorientation has ceased.

[0240] The term "alkyl" as used herein includes both branched and straight-chain saturated aliphatic hydrocarbon groups and has a specified number of carbon atoms, generally from 1 to 12 carbon atoms. For example, the term C1-C6 alkyl as used herein refers to an alkyl group having from 1 to 6 carbon atoms. When C0-Cn alkyl is used in conjunction with another group herein, taking (phenyl)C0-C4 alkyl as an example, the specified group is where the phenyl group is directly bonded by a single covalent bond (C0) or linked by an alkyl chain having a specified number of carbon atoms (in this case, from 1 to 4 carbon atoms). Examples of alkyl groups include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, 3-methylbutyl, tert-butyl, n-pentyl, and sec-pentyl.

[0241] The term "alkoxy" as used herein refers to a carbon atom having a specified number of carbon atoms connected by oxygen bridges, such as an "alkyl" atom as defined herein. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, 3-hexyloxy, and 3-methylpentoxy.

[0242] The term "heterocyclic alkyl" as used herein refers to a 5- to 8-membered saturated ring, a partially unsaturated ring, or an aromatic ring containing 1 to 4 heteroatoms selected from N, O, and S, with the remaining ring atom being carbon; or a 7- to 11-membered saturated ring, a partially unsaturated ring, or an aromatic heterocyclic system and a 10- to 15-membered tricyclic system containing at least one heteroatom selected from N, O, and S in a polycyclic system, and each ring in the polycyclic system containing at most about 4 heteroatoms independently selected from N, O, and S. Unless otherwise specified, the heterocycle may be attached to a group that substitutes at any heteroatom and carbon atom to produce a stable structure. When specified, the heterocycles described herein may be substituted at a carbon or nitrogen atom, provided that the resulting compound is stable. The nitrogen atom in the heterocycle may optionally be quaternized. Preferably, the total number of heteroatoms in the heterocyclic group is not greater than 4, and preferably the total number of S and O atoms in the heterocyclic group is not greater than 2, more preferably not greater than 1. Examples of heterocyclic groups include: pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, pyranyl, etc.

[0243] The term "aryl" or "heteroaryl" as used herein refers to a stable 5- or 6-membered monocyclic or polycyclic ring comprising 1 to 4, or preferably 1 to 3, heteroatoms selected from N, O, and S, with the remaining ring atom being carbon. When the total number of S and O atoms in the heteroaryl exceeds 1, these heteroatoms are not adjacent to each other. Preferably, the total number of S and O atoms in the heteroaryl is not greater than 2. Particularly preferred is that the total number of S and O atoms in the heteroaryl is not greater than 1. The nitrogen atom in the heterocycle may optionally be quaternized. When specified, these heteroaryls may also be substituted with carbon or non-carbon atoms or groups. Such substitution may include fusion with a 5- to 7-membered saturated cyclic group optionally comprising 1 or 2 heteroatoms independently selected from N, O, and S, thereby forming, for example, [1,3]dioxazolo[4,5-c]pyridyl. Examples of heteroaryl groups include, but are not limited to: pyridyl, indolyl, pyrimidinyl, pyridazinyl, pyrazinyl, imidazolyl, oxazolyl, furanyl, phenylthio, thiazolyl, triazolyl, tetrazolyl, isoxazolyl, quinolinyl, pyrroleyl, pyrazolyl, benzo[b]phenylthio, isoquinolinyl, quinazolinyl, quinoxalinyl, thiophene, isoindolyl, and 5,6,7,8-tetrahydroisoquinoline.

[0244] The terms “pharmaceutically acceptable salt” and “salt of a compound” used herein are interchangeable and both refer to derivatives of the disclosed compound, wherein the parent compound is prepared by the addition of a non-toxic acid or its base to form a salt. Examples of pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts: salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid. Other pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogen sulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentylpropionate, diglucuronate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucohepanoate, glyceryl phosphate, glucuronate, hemisulfate, heptaate, hexanoate, hydroiodate, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, dihydroxynaphthalate, pectate, persulfate, 3-phenylpropionate, phosphate, picrate, neopentanoate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium. Where appropriate, other pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl groups having 1 to 6 carbon atoms, sulfonate, and arylsulfonate.

[0245] The term "cancer" as used in this article includes, but is not limited to, the following cancers: Epidermoid oral cavity: buccal cavity, lip, tongue, mouth, pharynx; Cardiac: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyosarcoma, fibroma, lipoma, and teratoma; Lung: Bronchial carcinoma (squamous cell or epidermoid carcinoma, undifferentiated small cell carcinoma, undifferentiated large cell carcinoma, adenocarcinoma), honeycomb (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, hamartoma, mesothelioma; Gastrointestinal tract: esophagus (squamous cell carcinoma, laryngeal carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma), Gastrinoma, carcinoid tumor, vasoactive intestinal polypeptide tumor (vipoma), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, villonoma, hamartoma, leiomyoma), colon, colon-rectum, colorectal, rectum; genitourinary tract: kidney (adenocarcinoma, Wilms' tumor (nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminomatous seminoma, teratoma, embryonal carcinoma, teratoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroma) Adenoma, adenoma-like tumor, lipoma); Liver: Hepatocellular carcinoma, cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma, bile duct; Bone: Osteosarcoma, fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticular cell sarcoma), multiple myeloma, malignant giant cell tumor, chordoma, osteochronforma, benign chondroma, chondroblastoma, chondromyosarcoma-like fibroma, osteoid osteoma, and giant cell tumor; Nervous system: Skull (osteomas, Hemangioma, granuloma, xanthoma, osteitis deformans), meninges (meningioma, meningeal sarcoma, glioma), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germ cell tumor (pineal gland tumor), glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumor), spinal neurofibroma, meningioma, glioma, sarcoma; Gynecology: uterus (endometrial cancer), cervix (cervical cancer, pretumoral cervical lesions), ovary (ovarian cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma)), granulosa-theca cell tumor. Tumors), Sertoli-Leydig cell tumors, dysgerminomas, malignant teratomas), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonic rhabdomyosarcoma), fallopian tubes (cancer), breasts;Hematology: Blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative disorders, multiple myeloma, myelodysplastic syndromes), Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma) hairy cells; lymphoid diseases; Skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, keratoacanthoma, moles dysplastic nevi, lipoma, hemangioma, dermatofibroma, keloid, psoriasis; Thyroid: papillary thyroid carcinoma, follicular thyroid carcinoma; medullary thyroid carcinoma, undifferentiated thyroid carcinoma, type 2A multiple endocrine tumor, type 2B multiple endocrine tumor, familial medullary thyroid carcinoma, pheochromocytoma, paraganglioma; and Adrenal glands: neuroblastoma. Therefore, the term "cancer cell" as used herein includes cells affected by any of the above conditions.

[0246] Example

[0247] The present invention can be further described through the following embodiments, but the scope of the present invention is not limited to the following embodiments. Those skilled in the art will understand that various changes and modifications can be made to the present invention without departing from its spirit and scope.

[0248] All compounds were validated by UPLC-MS (waters) using LC-MS.

[0249] Column type: ACQUITY UPLC BEH C18 1.7μm; 2.1×50mm column.

[0250] Method A:5to 100% MeCN / H2O with 0.1% FA in 10min.

[0251] Method B:5to 95% MeCN / H2O with 0.1% FA in 6min.

[0252] Example 1: Synthesis of Compounds 01-36

[0253]

[0254] Scheme 1Synthesis of Compounds 01-36

[0255] Table 1. Schematic diagram of the substituent Ra in the intermediate compound of Scheme 1

[0256]

[0257]

[0258]

[0259] Following the synthetic route described above, Boc-D-2-aminobutyric acid (8 g, 39.37 mmol, 1.0 eq.), a methanol solution of methylamine (4.9 g, 47.24 mmol, 1.2 eq. 30–35% in methanol), NMI (11.3 g, 137.80 mmol, 3.5 eq.), and TCFH (12.1 g, 43.30 mmol, 1.1 eq.) were dissolved in acetonitrile (100 mL) and reacted overnight at room temperature. TLC analysis showed complete reaction of the starting material. After rotary evaporation to dryness, the product was dissolved in water, extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Column chromatography purification yielded the target compound DL1-2, a white solid, 7.9 g, in 82% yield.

[0260] UPLC-MS m / z:calcd for C 10 H 21 N2O3 + [M+H] + 217.15; found: 217.10; t R =2.369min

[0261] (Method A);

[0262]

[0263] tert-butyl(R)-(1-(methylamino)-1-oxobutan-2-yl)carbamate(2a)

[0264] Compound 2a (7.9 g, 36.53 mmol, 1.0 eq.) was dissolved in dichloromethane (100 mL), and dioxane hydrochloride (4 M, 30 mL) solution was added. The mixture was stirred overnight at room temperature. The reaction proceeded to completion by TLC. The mixture was filtered, the solid was dissolved in water, and the pH was adjusted to 8-9 with ammonia. The solution was extracted with dichloromethane, washed with saturated sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated to give the target compound 3a, 3.95 g of white solid, in 90% yield.

[0265] UPLC-MS m / z:calcd for C5H 13 N2O + [M+H] + 117.10; found: 117.15; t R = 1.552 min (Method A);

[0266]

[0267] (R)-2-amino-N-methylbutanamide(3a)

[0268] Compound 4a (2-hydroxy-5-bromoacetophenone) (20 g, 93.10 mmol, 1.0 eq.) was dissolved in anhydrous DMF (250 mL) at -10 °C and stirred for 30 min. Then, POCl3 (28.53 g, 186.02 mmol, 2.0 eq.) solution was slowly added dropwise at a temperature below 10 °C. The reaction was carried out overnight at room temperature under an argon atmosphere. TLC analysis confirmed complete reaction of the starting material. The reaction mixture was poured into ice water, the product was filtered, and washed with petroleum ether to obtain a crude product. Column chromatography purification yielded the target compound 5a, a pale yellow solid, 12.1 g, with a yield of 51%.

[0269] 1 H NMR (400MHz, DMSO-d6) δ10.09(s,1H),8.97(s,1H),8.18(d,J=2.4Hz,1H),8.04(dd,J=8.9,2.5Hz,1H),7.76(d,J=8.9Hz,1H)ppm.UPLC-MS m / z:calcd for C 10 H6BrO3 + [M+H] + 253.94; found: 254.87; t R = 3.207 min (Method A).

[0270]

[0271] 6-bromo-4-oxo-4H-chromene-3-carbaldehyde(5a)

[0272] An aqueous solution (75 mL) of NaClO2 (17.3 g, 191.27 mmol, 4.0 eq.) was added dropwise to a solution of 5a (12.1 g, 47.82 mmol, 1.0 eq.) in dichloromethane (200 mL) and NH2SO3H (23.22 g, 239.09 mmol, 5.0 eq.) in water (150 mL). The reaction was carried out overnight at 0 °C. TLC analysis showed that the reactants reacted completely. The organic phase was removed, and the aqueous phase was extracted with dichloromethane. The organic phases were then combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated, and finally washed with petroleum ether to obtain the crude product. Column chromatography purification yielded the target compound 6a, a pale yellow solid, 7.85 g, in a yield of 61%.

[0273] 1 H NMR (400MHz, DMSO-d6) δ10.09(s,1H),8.97(s,1H),8.18(d,J=2.4Hz,1H),8.04(dd,J=8.9,2.5Hz,1H),7.76(d,J=8.9Hz,1H)ppm.UPLC-MS m / z:calcd for C 10 H6BrO3 + [M+H] + 253.94; found: 254.87; t R = 3.207 min (Method A).

[0274]

[0275] 6-bromo-4-oxo-4H-chromene-3-carboxylic acid(6a)

[0276] Compounds 6a (2 g, 7.44 mmol, 1.0 eq.), 3a (864 mg, 7.44 mmol, 1.0 eq.), HATU (4.2 g, 11.16 mmol, 1.5 eq.), and DIPEA (2.9 g, 22.32 mmol, 3.0 eq.) were dissolved in anhydrous DMF (30 mL) and reacted overnight at room temperature under argon protection. TLC analysis showed that the starting material reacted completely. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Column chromatography purification yielded the target compound 7a, a pale yellow solid, 2.19 g, in 80% yield.

[0277] 1 H NMR (400MHz, DMSO-d6) δ9.08(s,1H),9.04(s,1H),8.16(t,J=2.1Hz,1H),8.02(d,J=8.9Hz,1H),7.73(d,J=8.9Hz,1H)ppm.UPLC-MS m / z:calcd for C 10 H6BrO4 + [M+H] + 269.94; found: 270.01; t R = 3.760 min (Method A).

[0278]

[0279] (R)-6-bromo-N-(1-(methylamino)-1-oxobutan-2-yl)-4-oxo-4H-chromene-3-carboxamide(7a)

[0280] Compound 9a (10 g, 35.03 mmol, 1.0 eq.) was dissolved in dichloromethane (150 mL), and 8a-8z or 8aa-8aj (2.2-12.7 g, 70.05 mmol, 2.0 eq.) was added. The reaction was carried out at room temperature for 8 h. TLC analysis showed that the starting material reacted completely. The reaction solution was washed successively with 0.5 M HCl aqueous solution and saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Column chromatography purification yielded the target compounds 10a-10z and 10aa-10aj, 5-10.6 g of white solid, with a yield of 65-85%.

[0281] 1 H NMR (400MHz, DMSO-d6) δ11.86(dd,J=14.6,8.3Hz,1H),8.52(d,J=14.6Hz,1H),8.31(d,J=4.6Hz,1H),7.93(d,J=2.4Hz,1H),7.83–7.7 2(m,1H),7.27(dd,J=16.6,8.7Hz,1H),4.49–4.16(m,1H),2.62(d,J=4.5Hz,3H),1.96–1.66(m,2H),0.80(t,J=7.5Hz,3H)ppm.UPLC-MS m / z:calcd for C 15 H 16 BrN2O4 + [M+H] + 368.02; found: 368.13; t R = 4.281 min (Method A).

[0282]

[0283] Intermediate compounds 10a-10z and 10aa-10aj

[0284] Compounds 10a-10z or 10aa-10aj (5-10.6 g, 29.76 mmol, 1.0 eq.), bis(pinacol)diboron (11.34 g, 44.64 mmol, 1.5 eq.), potassium acetate (11.69 g, 119.03 mmol, 4.0 eq.), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (2.18 g, 2.98 mmol, 0.1 eq.) were added to a 1,4-dioxane (150 mL) solution. The reaction system was stirred overnight at 95 °C under argon protection. TLC analysis showed that the reaction proceeded completely. The reaction mixture was diluted with water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. After drying with anhydrous sodium sulfate, the crude product was concentrated and purified by column chromatography to obtain the target compounds 11a-11z and 11aa-11aj, in 4-9.36 g brick-red solids, with a yield of 50-78%.

[0285]

[0286] Intermediate compounds 11a-11z and 11aa-11aj

[0287] Compounds 11a-11z or 11aa-11aj (40-80 mg, 0.22 mmol, 1.0 eq.), compound 7a (86 mg, 0.22 mmol, 1.0 eq.), tetra-triphenylphosphine palladium (25 mg, 0.02 mmol, 0.1 eq.), and potassium carbonate (88 mg, 0.64 mmol, 3.0 eq.) were dissolved in 1,4-dioxane / water (4 / 1, 5 mL). The reaction mixture was refluxed overnight at 80 °C under argon protection. TLC analysis showed that the reactants reacted completely. The mixture was diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Prep-HPLC separation yielded the target compounds 01-36, 20-44 mg of white solid, in a yield of 25-40%.

[0288]

[0289] Compound 01-36

[0290] The substituent Ra in compounds 01-36 are shown in Table 2.

[0291] Table 2 shows the structural diagram of substituent Ra in compound 01-36 of Scheme 1.

[0292]

[0293]

[0294] Example 2: Synthesis of compounds 37-46

[0295] Scheme 2Synthesis of Compounds 37-46

[0296] Table 3. Schematic diagram of the substituent Rb in the intermediate compound of Scheme 2.

[0297]

[0298] Following the aforementioned synthetic pathway, compound 8ak-8at (5-10 g, 30 mmol, 1.0 eq.) was dissolved in dichloromethane (150 mL), and 9b (12.0 g, 60 mmol, 2.0 eq.) was added. The reaction was carried out at room temperature for 8 h. TLC analysis showed that the reaction proceeds were completely reacted. The reaction solution was washed successively with 0.5 M HCl aqueous solution and saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Column chromatography purification yielded the target compound 10ak-10at, 5-10 g of white solid, with a yield of 65-80%.

[0299]

[0300] Intermediate compound 10ak-10at

[0301] Compound 10ak-10at (5-10.0 g, 29.76 mmol, 1.0 eq.), bis(pinacol)diboron (11.34 g, 44.64 mmol, 1.5 eq.), potassium acetate (11.69 g, 119.03 mmol, 4.0 eq.), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (2.18 g, 2.98 mmol, 0.1 eq.) were added to a 1,4-dioxane (150 mL) solution. The reaction mixture was stirred overnight at 95 °C under argon protection. TLC analysis showed complete reaction of the starting materials. The reaction mixture was diluted with water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Column chromatography purification yielded the target compound 11ak-11at, a brick-red solid, 4-9.5 g, in yields of 50-75%.

[0302]

[0303] Intermediate compound 11ak-11at

[0304] Compounds 11ak-11at (40-80 mg, 0.22 mmol, 1.0 eq.), compound 7a (86 mg, 0.22 mmol, 1.0 eq.), tetra-triphenylphosphine palladium (25 mg, 0.02 mmol, 0.1 eq.), and potassium carbonate (88 mg, 0.64 mmol, 3.0 eq.) were dissolved in 1,4-dioxane / water (4 / 1, 5 mL). The reaction mixture was refluxed overnight at 80 °C under argon protection. TLC analysis showed that the reactants reacted completely. The mixture was diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Prep-HPLC separation yielded target compounds 37-46, 20-44 mg of white solid, in yields of 25-45%.

[0305]

[0306] Compound 37-46

[0307] The substituents Rb in compounds 37-46 are shown in Table 4.

[0308] Table 4. Schematic diagram of the Rb substituents 37-46 in compound 2 of scheme 4.

[0309]

[0310] Example 3 Compounds 47-52

[0311]

[0312] Scheme 3Synthesis of Compounds 47-52

[0313] Table 5-1 Schematic diagram of the substituent Rc structure of intermediate compound in Scheme 3

[0314]

[0315] Table 5-2 Schematic diagram of substituents Rd and Rc in intermediate compounds of Scheme 3

[0316]

[0317]

[0318] Compound 9c-9g (5-10g, 30mmol, 1.0eq.) was dissolved in dichloromethane (150mL), and 8a or 8i (10-12.0g, 60mmol, 2.0eq.) was added. The reaction was carried out at room temperature for 8h. TLC analysis showed that the starting material reacted completely. The reaction solution was washed successively with aqueous HCl solution (0.5M) and saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Column chromatography purification yielded the target compound 10au-10az, 5-11g of white solid, in a yield of 65-85%.

[0319]

[0320] Intermediate compound 10au-10az

[0321] Compound 10au-10az (5-11 g, 29.76 mmol, 1.0 eq.), bis(pinacol)diboron (11.34 g, 44.64 mmol, 1.5 eq.), potassium acetate (11.69 g, 119.03 mmol, 4.0 eq.), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (2.18 g, 2.98 mmol, 0.1 eq.) were added to a 1,4-dioxane (150 mL) solution. The reaction mixture was stirred overnight at 95 °C under argon protection. TLC analysis showed complete reaction of the starting materials. The reaction mixture was diluted with water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Column chromatography purification yielded the target compound 11au-11az, 4-9.5 g of brick-red solid, in yields of 50-78%.

[0322]

[0323] Intermediate compound 11au-11az

[0324] Compounds 11au-11az (40-80 mg, 0.22 mmol, 1.0 eq.), compound 7a (86 mg, 0.22 mmol, 1.0 eq.), tetra-triphenylphosphine palladium (25 mg, 0.02 mmol, 0.1 eq.), and potassium carbonate (88 mg, 0.64 mmol, 3.0 eq.) were dissolved in 1,4-dioxane / water (4 / 1, 5 mL). The reaction mixture was refluxed overnight at 80 °C under argon protection. TLC analysis showed that the reactants reacted completely. The mixture was diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Prep-HPLC separation yielded target compounds 47-52, 20-44 mg of white solid, in yields of 35-50%.

[0325]

[0326] Compound 47-52

[0327] The substituents Rd and Rc in compounds 47-52 are shown in Table 6.

[0328] Table 6 shows the structural diagrams of substituents Rd and Rc in compounds 47-52.

[0329]

[0330] Example 4 Synthesis of compounds 53-54

[0331]

[0332] Scheme 4Synthesis of Compounds 53-54

[0333] Table 7. Structural diagram of the substituent Re in the intermediate compound of Scheme 4.

[0334]

[0335] Compounds benzylamine (266 mg, 2.49 mmol, 1.0 eq.), 9h-9i (500-575 mg, 2.49 mmol, 1.0 eq.), HATU (1420 mg, 3.74 mmol, 1.5 eq.), and DIPEA (965 mg, 7.47 mmol, 3.0 eq.) were dissolved in anhydrous DMF (10 mL) and reacted overnight at room temperature under argon protection. TLC analysis showed the reactants reacted completely. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Column chromatography yielded the target compound 10ba-10bb, a white solid, 600-707 mg, in a yield of 75-80%.

[0336]

[0337] Intermediate compound 10ba-10bb

[0338] Compound 10ba-10bb (300-320 mg, 1.03 mmol, 1.0 eq.), bis(pinacol)diboron (410 mg, 1.6 mmol, 1.5 eq.), potassium acetate (400 mg, 4 mmol, 4.0 eq.), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (75 mg, 0.1 mmol, 0.1 eq.) were added to a 1,4-dioxane (150 mL) solution. The reaction mixture was stirred overnight at 95 °C under argon protection. TLC analysis showed complete reaction of the starting material. The reaction mixture was diluted with water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Column chromatography purification yielded the target compound 11ba-11bb, 200-230 mg of brick-red solid, in a yield of 50-78%.

[0339]

[0340] Intermediate compound 11ba-11bb

[0341] Compounds 11ba-11bb (50-70 mg, 0.20 mmol, 1.0 eq.), compound 7a (80 mg, 0.20 mmol, 1.0 eq.), tetraphenylphosphine palladium (25 mg, 0.02 mmol, 0.1 eq.), and potassium carbonate (83 mg, 0.60 mmol, 3.0 eq.) were dissolved in 1,4-dioxane / water (4 / 1, 5 mL). The reaction mixture was refluxed overnight at 80 °C under argon protection. TLC analysis showed that the reactants reacted completely. The mixture was diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Prep-HPLC separation yielded target compounds 53-54, 20-44 mg of white solid, in a yield of 25-40%.

[0342]

[0343] Compound 53-54

[0344] The substituents Re in compounds 53-54 are shown in Table 8.

[0345] Table 8 shows the structural diagrams of the substituent Re in compounds 53-54.

[0346]

[0347] Example 5: Synthesis of compounds 55-63

[0348]

[0349] Scheme 5 Synthesis of Compounds 55-63

[0350] Table 9. Structural diagram of the substituent Rf in the intermediate compound of Scheme 5.

[0351]

[0352]

[0353] Compounds 6b-6j (2-2.5 g, 7.44 mmol, 1.0 eq.), 3a (864 mg, 7.44 mmol, 1.0 eq.), HATU (4.2 g, 11.16 mmol, 1.5 eq.), and DIPEA (2.9 g, 22.32 mmol, 3.0 eq.) were dissolved in anhydrous DMF (30 mL) and reacted overnight at room temperature under argon protection. TLC analysis showed that the reactants reacted completely. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Column chromatography purification yielded the target compound 7b-7j, a pale yellow solid, 1.5-2.2 g, in 50-80% yield.

[0354]

[0355] Intermediate compound 7b-7j

[0356] Compound 11a (80 mg, 0.20 mmol, 1.0 eq.), compounds 7b-7j (50-70 mg, 0.20 mmol, 1.0 eq.), tetra-triphenylphosphine palladium (25 mg, 0.02 mmol, 0.1 eq.), and potassium carbonate (83 mg, 0.60 mmol, 3.0 eq.) were dissolved in 1,4-dioxane / water (4 / 1, 5 mL). The reaction mixture was refluxed overnight at 80 °C under argon protection. TLC analysis showed that the reactants reacted completely. The mixture was diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Prep-HPLC separation yielded target compounds 55-63, 20-44 mg of white solid, in yields of 25-40%.

[0357]

[0358] Compound 55-63

[0359] The structural schematic diagrams of the substituent Rf in compounds 55-63 are shown in Table 10.

[0360] Table 10 shows the structural diagrams of the substituent Rf in compounds 55-63.

[0361]

[0362] Example 6 Synthesis of compounds 64-73

[0363]

[0364] Scheme 6 Synthesis of Compounds 64-73 Table 11 Schematic diagram of the substituent Rg in the intermediate compound of Scheme 6

[0365]

[0366] Compound 1b-1k (7.5-9 g, 39.37 mmol, 1.0 eq.), a methanol solution of methylamine (4.9 g, 47.24 mmol, 1.2 eq. 30-35% in methanol), NMI (11.3 g, 137.80 mmol, 3.5 eq.), and TCFH (12.1 g, 43.30 mmol, 1.1 eq.) were dissolved in acetonitrile (100 mL) and reacted overnight at room temperature. TLC analysis showed the reactants were completely reacted. After rotary evaporation, the solution was dissolved in water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. After drying over anhydrous sodium sulfate, the crude product was concentrated and purified by column chromatography to give the target compound 2b-2k as a white solid (5-7.5 g, yield 50-82%).

[0367]

[0368] Intermediate compound 2b-2k

[0369] Compound 2b-2k (5-7.5 g, 36.53 mmol, 1.0 eq.) was dissolved in dichloromethane (100 mL), and dioxane hydrochloride (4 M, 30 mL) solution was added. The mixture was stirred overnight at room temperature. The reaction proceeded to completion by TLC. The mixture was filtered, and the solid was dissolved in water. The pH was adjusted to 8-9 with ammonia, extracted with dichloromethane, washed with saturated sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated to give the target compound 3b-3k as a white solid, 2-4 g, in 85-90% yield.

[0370]

[0371] Intermediate compound 3b-3k

[0372] Compounds 6a (2 g, 7.44 mmol, 1.0 eq.), 3b-3k (0.8-1.0 g, 7.44 mmol, 1.0 eq.), HATU (4.2 g, 11.16 mmol, 1.5 eq.), and DIPEA (2.9 g, 22.32 mmol, 3.0 eq.) were dissolved in anhydrous DMF (30 mL) and reacted overnight at room temperature under argon protection. TLC analysis showed the reactants reacted completely. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Column chromatography purification yielded the target compound 7k-7t, a pale yellow solid, 1.5-2.5 g, in 70-80% yield.

[0373]

[0374] Intermediate compound 7k-7t

[0375] Compound 11a (80 mg, 0.20 mmol, 1.0 eq.), compound 7k-7t (50-70 mg, 0.20 mmol, 1.0 eq.), tetra-triphenylphosphine palladium (25 mg, 0.02 mmol, 0.1 eq.), and potassium carbonate (83 mg, 0.60 mmol, 3.0 eq.) were dissolved in 1,4-dioxane / water (4 / 1, 5 mL). The reaction mixture was refluxed overnight at 80 °C under argon protection. TLC analysis showed that the reactants reacted completely. The mixture was diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Prep-HPLC separation yielded target compounds 64-73, 20-50 mg of white solid, in yields of 25-45%.

[0376]

[0377] Compound 64-73

[0378] The substituents Re in compounds 64-73 are shown in Table 12.

[0379] Table 12 shows the structural diagrams of the substituent Rg in compounds 64-73.

[0380]

[0381] Example 7 Synthesis of compounds 75-81

[0382]

[0383] Scheme 7Synthesis of Compounds 75-81

[0384] Table 13 Schematic diagram of the substituent Rh in intermediate compound of Scheme 7

[0385]

[0386] Compounds Boc-D-2-aminobutyric acid (0.8 g, 4 mmol, 1.0 eq.), 0b-0h (0.5-1 g, 4.8 mmol, 1.2 eq.), NMI (1.2 g, 14 mmol, 3.5 eq.), and TCFH (1.2 g, 4.4 mmol, 1.1 eq.) were dissolved in acetonitrile (10 mL) and reacted overnight at room temperature. TLC analysis showed that the reaction proceeded completely. After rotary evaporation to dryness, the product was dissolved in water, extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Column chromatography purification yielded the target compound 2l-2r as a white solid, 0.5-0.8 g, in a yield of 60-75%.

[0387]

[0388] Intermediate compound 2l-2r

[0389] Compound 2l-2r (0.5-0.8 g, 3 mmol, 1.0 eq.) was dissolved in dichloromethane (10 mL), and dioxane hydrochloride (4 M, 3 mL) solution was added. The mixture was stirred overnight at room temperature. The reaction proceeded to completion by TLC. The mixture was filtered, the solid was dissolved in water, and the pH was adjusted to 8-9 with ammonia. The solution was extracted with dichloromethane, washed with saturated sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated to give the target compound 3l-3r as a white solid (0.2-0.4 g, yield 80-90%).

[0390]

[0391] Intermediate compound 3l-3r

[0392] Compounds 6a (0.2 g, 0.75 mmol, 1.0 eq.), 3a (0.2–0.4 g, 0.75 mmol, 1.0 eq.), HATU (0.4 g, 1 mmol, 1.5 eq.), and DIPEA (0.3 g, 2.25 mmol, 3.0 eq.) were dissolved in anhydrous DMF (5 mL) and reacted overnight at room temperature under argon protection. TLC analysis showed the reactants reacted completely. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Column chromatography purification yielded the target compound 7u-7aa, a pale yellow solid, 0.2–0.3 g, in 70–80% yield.

[0393]

[0394] Intermediate compound 7u-7aa

[0395] Compound 11a (80 mg, 0.20 mmol, 1.0 eq.), compound 7u-7aa (50-70 mg, 0.20 mmol, 1.0 eq.), tetra-triphenylphosphine palladium (25 mg, 0.02 mmol, 0.1 eq.), and potassium carbonate (83 mg, 0.60 mmol, 3.0 eq.) were dissolved in 1,4-dioxane / water (4 / 1, 5 mL). The reaction mixture was refluxed overnight at 80 °C under argon protection. TLC analysis showed that the reactants reacted completely. The mixture was diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Prep-HPLC separation yielded target compounds 75-81, 20-50 mg of white solid, in yields of 25-45%.

[0396]

[0397] Compound 75-81

[0398] The substituents Rh in compounds 75-81 are shown in Table 14.

[0399] Table 14 shows the structural diagrams of the substituent Rh in compounds 75-81.

[0400]

[0401] Example 8 Synthesis of Compound 74

[0402]

[0403] Scheme 8Synthesis of Compounds 74

[0404] Compound 6-bromobenzo[d]oxazol-2(3H)-one (500 mg, 2.34 mmol, 1.0 eq.) and potassium carbonate (485 mg, 3.51 mmol, 1.5 eq.) were dissolved in a suspension of DMF and stirred in an ice bath for 10 min. Tert-butyl bromoacetate (478 mg, 2.45 mmol, 1.05 eq.) was added dropwise to the mixture, and the reaction mixture was stirred at room temperature for 3 h. TLC analysis showed that the starting material reacted completely. The reaction mixture was dissolved in water, extracted with ethyl acetate, washed with saturated sodium chloride solution of the organic phase, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Column chromatography purification yielded the target compound 12a as a white solid (590 mg, 77% yield).

[0405] UPLC-MS m / z:calcd for C 13 H 15 BrNO4 + [M+H] + 329.01; found: 329.05; t R = 3.060 min (Method B).

[0406]

[0407] tert-butyl 2-(5-bromo-2-oxobenzo[d]oxazol-3(2H)-yl)acetate(12a)

[0408] Compound 12a (590 mg, 1.80 mmol, 1.0 eq.) was dissolved in TFA and reacted at room temperature for 3 h. TLC analysis showed that the reaction proceeded completely. After evaporating the solvent, the crude product was obtained, which was purified by column chromatography to give the target compound 13a as a brown solid (445 mg), with a yield of 91%.

[0409] UPLC-MS m / z:calcd for C9H7BrNO4 + [M+H] + 272.95; found: 272.98; t R = 2.117 min (Method B).

[0410]

[0411] 2-(5-bromo-2-oxobenzo[d]oxazol-3(2H)-yl)acetic acid(13a)

[0412] Compound 13a (445 mg, 1.64 mol, 1.0 eq.), a methanol solution of methylamine (170 mg, 1.64 mmol, 1.0 eq. 30–35% in methanol), NMI (0.5 g, 5.74 mmol, 3.5 eq.), and TCFH (0.5 g, 1.80 mmol, 1.1 eq.) were dissolved in acetonitrile (10 mL) and reacted overnight at room temperature. TLC analysis showed the reactants reacted completely. After rotary evaporation to dryness, the solution was dissolved in water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. After drying over anhydrous sodium sulfate, the crude product was concentrated and purified by column chromatography to give the target compound 14a as a white solid (335 mg, yield 72%).

[0413] UPLC-MS m / z:calcd for C 10 H10 BrN2O3 + [M+H] + 285.98; found: 286.93; t R = 2.043 min (Method B).

[0414]

[0415] (102)2-(5-bromo-2-oxobenzo[d]oxazol-3(2H)-yl)acetic acid(14a)

[0416] Compound 11a (80 mg, 0.20 mmol, 1.0 eq.), compound 14a (60 mg, 0.20 mmol, 1.0 eq.), tetrakis(triphenylphosphine)palladium (25 mg, 0.02 mmol, 0.1 eq.), and potassium carbonate (83 mg, 0.60 mmol, 3.0 eq.) were dissolved in 1,4-dioxane / water (4 / 1, 5 mL). The reaction mixture was refluxed overnight at 80 °C under argon protection. TLC analysis showed that the reactants reacted completely. The mixture was diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Prep-HPLC separation yielded target compound 74 as a white solid (30 mg, 35% yield).

[0417]

[0418] 2-(5-(3-(N-benzylsulfamoyl)-4-methoxyphenyl)-2-oxobenzo[d]oxazol-3(2H)-yl)-N-methylacetamide(74)

[0419] Example 9 Synthesis of Compound 82

[0420]

[0421] Scheme 9Synthesis of Compounds 82

[0422] Compound Boc-4-bromo-L-beta-phenylalanine (300 mg, 0.87 mol, 1.0 eq.), a methanol solution of methylamine (91 mg, 0.87 mmol, 1.0 eq. 30–35% in methanol), NMI (0.27 g, 3 mmol, 3.5 eq.), and TCFH (0.24 g, 0.88 mmol, 1.1 eq.) were dissolved in acetonitrile (5 mL) and reacted overnight at room temperature. TLC analysis showed the reactants reacted completely. After rotary evaporation to dryness, the solution was dissolved in water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. After drying with anhydrous sodium sulfate, the crude product was concentrated and purified by column chromatography to give the target compound 17a as a white solid (252 mg, yield 81%).

[0423] UPLC-MS m / z:calcd for C 15 H 22 BrN2O3 + [M+H] + 358.07; found: 358.13; t R =2.246min

[0424] (Method B)

[0425]

[0426] tert-butyl(S)-(1-(4-bromophenyl)-3-(methylamino)-3-oxopropyl)carbamate(17a)

[0427] Compound 17a (252 mg, 1.65 mmol, 1.0 eq.) was dissolved in dichloromethane (5 mL), and dioxane hydrochloride (4 M, 3 mL) solution was added. The mixture was stirred overnight at room temperature. The reaction proceeded to completion by TLC. The mixture was filtered, the solid was dissolved in water, and the pH was adjusted to 8-9 with ammonia. The solution was extracted with dichloromethane, washed with saturated sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated to give the target compound 18a as a white solid (136 mg, yield 75%).

[0428] UPLC-MS m / z:calcd for C 10 H 14 BrN2O + [M+H] + 258.02; found: 258.12; t R =1.977min

[0429] (Method B)

[0430]

[0431] (S)-3-amino-3-(4-bromophenyl)-N-methylpropanamide(18a)

[0432] Compound 11a (80 mg, 0.20 mmol, 1.0 eq.), compound 6a (54 mg, 0.20 mmol, 1.0 eq.), tetrakis(triphenylphosphine)palladium (25 mg, 0.02 mmol, 0.1 eq.), and potassium carbonate (83 mg, 0.60 mmol, 3.0 eq.) were dissolved in 1,4-dioxane / water (4 / 1, 5 mL). The reaction mixture was refluxed overnight at 80 °C under argon protection. TLC analysis showed that the reactants reacted completely. The mixture was diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Prep-HPLC separation yielded target compound 19a as a white solid (55 mg, yield 56%).

[0433]

[0434] 6-(3-(N-benzylsulfamoyl)-4-methoxyphenyl)-4-oxo-4H-chromene-3-carboxylic acid(19a)

[0435] Compounds 19a (55 mg, 0.12 mmol, 1.0 eq.), 18a (26 mg, 0.12 mmol, 1.0 eq.), HATU (68 mg, 0.18 mmol, 1.5 eq.), and DIPEA (47 g, 0.36 mmol, 3.0 eq.) were dissolved in anhydrous DMF (2 mL) and reacted overnight at room temperature under argon protection. TLC analysis showed that the reactants reacted completely. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Prep-HPLC separation yielded the target compound 82 as a white solid (55 mg), with a yield of 65%.

[0436]

[0437] (S)-6-(3-(N-benzylsulfamoyl)-4-methoxyphenyl)-N-(1-(4-bromophenyl)-3-(methylamino)-3-oxop

[0438] ropyl)-4-oxo-4H-chromene-3-carboxamide(82)

[0439] Example 10 Synthesis of compounds 83-84

[0440]

[0441] Scheme 10Synthesis of Compounds 83

[0442] Intermediate 20a (5 g, 24.63 mmol, 1.0 eq.), imidazole (2 g, 29.56 mmol, 1.2 eq.), and potassium carbonate (4.1 g, 29.56 mmol, 1.2 eq.) were dissolved in DMF (20 mL), and the reaction mixture was incubated overnight at 90 °C. TLC analysis showed that the reaction proceeded completely. The mixture was diluted with water and extracted with ethyl acetate. After drying over anhydrous sodium sulfate and concentration, the solution was purified by column chromatography to give the target compound 21a as a pale yellow solid (3.4 g, yield 55%).

[0443] UPLC-MS calculated for C 10 H8BrN2O + [M+H] + :250.98; found:251.97.UPLC-retention time:1.5min. 1 H NMR (400MHz, DMSO-d6) δ10.05(s,1H),8.35(s,1H),8.16–7.81(m,2H),7.74(d,J=8.2Hz,1H),7.52(s,1H),7.14(s,1H).

[0444]

[0445] 3-bromo-4-(1H-imidazol-1-yl)benzaldehyde(21a)

[0446] Intermediate 21a (500 mg, 2 mmol, 1.0 eq.), intermediate 11a (810 mg, 2 mmol, 1.0 eq.), tetratetraphenylphosphine palladium (15 mg, 2 mmol, 0.02 eq.), and potassium carbonate (830 mg, 6 mmol, 3.0 eq.) were dissolved in 1,4-dioxane / water (5 mL, 4 / 1). The reaction mixture was refluxed overnight at 80 °C under argon protection. TLC analysis showed that the reaction proceeded completely. The mixture was diluted with water and extracted with ethyl acetate. After drying over anhydrous sodium sulfate and concentration, the solution was purified by column chromatography to give target compound 22a as a white solid (580 mg, 65% yield).

[0447] UPLC-MS calculated for C 24 H 22 N3O4S + [M+H] + :448.14found:448.05.UPLC-retention time:2.4min. 1 H NMR (400MHz, DMSO-d6) δ10.15(s,1H),8.08–8.00(m,2H),7.89(t,J=6.4Hz,1H),7.72(d,J=8.0Hz,1H),7.65(s,1H) ,7.49(d,J=2.3Hz,1H),7.27–7.13(m,7H),7.05(d,J=8.7Hz,1H),6.98(s,1H),3.96(d,J=6.4Hz,2H),3.80(s,3H).

[0448]

[0449] N-benzyl-5'-formyl-2'-(1H-imidazol-1-yl)-4-methoxy-[1,1'-biphenyl]-3-sulfonamide(22a)

[0450] Intermediate 22a (50 mg, 0.11 mmol, 1.0 eq.) and intermediate 16a (16 mg, 0.11 mmol, 1.0 eq.) were dissolved in 1,2-dichloroethane (2 mL). One drop of acetic acid was added, and the mixture was stirred at room temperature for 30 min. Then, sodium triacetoxyborohydride (46 mg, 0.22 mmol, 2.0 eq.) was added, and the mixture was stirred at room temperature for 3 h. TLC analysis showed that the reaction proceeded completely. The mixture was diluted with water, the reaction was quenched with sodium bicarbonate solution, and extracted with ethyl acetate. After drying over anhydrous sodium sulfate and concentration, the solution was separated by Prep-HPLC to obtain the target compound 83 as a white solid (36 mg, yield 55%).

[0451]

[0452] (S)-2-(((3'-(N-benzylsulfamoyl)-6-(1H-imidazol-1-yl)-4'-methoxy-[1,1'-biphenyl]-3-

[0453] yl)methyl)amino)-N-methylhexanamide(83)

[0454]

[0455] Scheme 11Synthesis of Compounds 84

[0456] Intermediate 22a (50 mg, 0.11 mmol, 1.0 eq.) and intermediate 23b (22 mg, 0.11 mmol, 1.0 eq.) were dissolved in 1,2-dichloroethane (2 mL). One drop of acetic acid was added, and the mixture was stirred at room temperature for 30 min. Then, sodium triacetoxyborohydride (46 mg, 0.22 mmol, 2.0 eq.) was added, and the mixture was stirred at room temperature for 3 h. TLC analysis showed that the reaction proceeded completely. The mixture was diluted with water, the reaction was quenched with sodium bicarbonate solution, and extracted with ethyl acetate. After drying over anhydrous sodium sulfate and concentration, the solution was separated by Prep-HPLC to obtain the target compound 84 as a white solid (30 mg, yield 45%).

[0457]

[0458] (S)-2-(3-(((3'-(N-benzylsulfamoyl)-6-(1H-imidazol-1-yl)-4'-methoxy-[1,1'-biphenyl]-3-yl)methyl)amino)-2-oxoazepan-1-yl)-N-methylacetamide(84)

[0459] Table 15 Summary of information for compounds 01-84

[0460]

[0461]

[0462]

[0463]

[0464]

[0465]

[0466]

[0467]

[0468]

[0469]

[0470]

[0471]

[0472]

[0473]

[0474]

[0475]

[0476]

[0477]

[0478]

[0479]

[0480]

[0481]

[0482]

[0483]

[0484]

[0485]

[0486]

[0487]

[0488]

[0489]

[0490]

[0491]

[0492] Table 16 Summary of the activities of compounds 01-84

[0493]

[0494]

[0495]

[0496]

[0497]

[0498]

[0499]

[0500]

[0501]

[0502]

[0503]

[0504]

[0505]

[0506]

[0507]

[0508]

[0509] A:1-10μM; B:10-50μM; C:50-100μM; D:>100μM

[0510] Example 11: Surface Plasmon Resonance (SPR) Technique for Detecting Compound Affinity

[0511] SPR was used to detect the affinity of compounds Apcin (01, 64, 66, 83, 84) for Cdc20. The CM5 chip was installed and operated according to the Biacore T200 standard operating procedure. Purified Cdc20 protein was coupled to the chip using an amino-coupling kit. Compounds were serially diluted and then analyzed to obtain a series of SPR binding data. The small molecule binding time was set to 60 s, the dissociation time to 120 s, and the flow rate to 30 μL / min. Data analysis was performed using Biacore T200 analysis software. The experiment was repeated three times, and the purity of Cdc20 protein was above 90% (e.g., ...). Figure 1 (As shown).

[0512] Example 12 Inhibitory Activity of Androgen-Dependent Prostate Cancer Cell Proliferation

[0513] Cell proliferation after compound treatment was assessed using the WST-1 assay. MCF-7 (DMEM + 10% FBS), J82 (MEM + 10% FBS), HepG2 (MEM + 10% FBS), and PC3 (Ham's F-12K + 10% FBS) cells were passaged and seeded into 96-well cell culture plates at 8 × 10⁸ cells per well. 3 Cells were collected, and after 24 hours, different concentrations of Apcin (0, 3.125, 6.25, 12.5, 25, 50, 100 μM) were added to the cells. The cells were incubated at 37°C in a 5% CO2 incubator for 4-7 days. Following the kit instructions, the absorbance of each well at 450 nm was measured using a microplate reader (zeroed with a blank control). Two replicates were used for each group. Statistical analysis and plotting were performed using GraphPad Prism 8 software, and the IC50 was calculated. 50 Value (e.g.) Figure 2 (As shown).

[0514] Figure 1 , Figure 2 The specific results are shown in Table 17:

[0515] Table 17 Results of Cdc20 ligand activity

[0516]

[0517] Example 13 Synthesis of PROTACs molecules

[0518] These are PROTAC molecules designed using formula I and Apcin-A, respectively. The structures of Apcin and Apcin-A are as follows:

[0519]

[0520] The synthetic methods and routes for some of IIIa, IIIb, IVa, and IVb are as follows;

[0521]

[0522] Scheme 12Synthesis of PROTCs IIIa(MJC142-149,n=1-8)

[0523] AR ligand AR-1 (50 mg, 1.0 eq.), linkers of different lengths L1 (20-32 mg, 1.1 eq.), HATU (48 mg, 1.2 eq.), and DIEA (41 mg, 3.0 eq.) were dissolved in DMF (2 mL), and the reaction mixture was stirred overnight at room temperature. TLC analysis showed that the reaction proceeded completely. The mixture was diluted with water and extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated. The crude product was dissolved in EA (2 mL), and 0.5 mL of ethyl acetate hydrochloride solution was added dropwise. The mixture was stirred at room temperature for 2 hours to remove Boc. After the reaction was complete, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to obtain intermediate AR-L1 with linkers of different lengths, 25-35 mg of white solid, with a two-step yield of 47-65%.

[0524] Intermediate AR-L1 (25-35 mg, 1.0 eq.), Cdc20 ligand 19a (24-32 mg, 1.1 eq.), HATU (24-35 mg, 1.2 eq.), and DIEA (18-25 mg, 3.0 eq.) were dissolved in DMF (2 mL), and the reaction system was stirred overnight at room temperature. TLC analysis showed that the reaction proceeds were completely reacted. The reaction solution was purified by Prep-HPLC to obtain AR degrading agent MJC142-149, a white solid, at a yield of 30-55% (15-26 mg).

[0525]

[0526] Scheme 13Synthesis of PROTCs IIIa(MJC152-159,n=1-8)

[0527] Intermediate AR-L1 (25-35 mg, 1.0 eq.), Cdc20 ligand Ia (24-32 mg, 1.1 eq.), HATU (26-40 mg, 1.2 eq.), and DIEA (18-25 mg, 3.0 eq.) were dissolved in DMF (2 mL), and the reaction system was stirred overnight at room temperature. TLC analysis showed that the reaction proceeds were completely reacted. The reaction solution was purified by Prep-HPLC to obtain AR degrading agent MJC152-159, a white solid, at a yield of 28-50%, at a concentration of 10-25 mg.

[0528]

[0529] Scheme 14 Synthesis of PROTCs IIIb(MJC122-130,n=2-9)

[0530] JQ1 (1000 mg) was dissolved in formic acid (5 mL), and stirred at room temperature for 24 hours. The reaction was confirmed to be complete by TLC. The formic acid was removed by concentration to give 870 mg of JQ-acid, a pale yellow solid, with a yield of 99%.

[0531] JQ-acid (35 mg, 1.0 eq.), linkers L2 of different lengths (12-25 mg, 1.1 eq.), HATU (35 mg, 1.2 eq.), and DIEA (34 mg, 3.0 eq.) were dissolved in DMF (2 mL), and the reaction mixture was stirred overnight at room temperature. TLC analysis showed that the reaction proceeded completely. The mixture was diluted with water and extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated. The crude product was dissolved in EA (2 mL), and 0.5 mL of ethyl acetate hydrochloride solution was added dropwise. The mixture was stirred at room temperature for 2 hours to remove Boc. After the reaction was complete, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to obtain intermediate JQ-L2 with linkers of different lengths, 23-33 mg of white solid, with a two-step yield of 44-66%.

[0532] Intermediate JQ-L2 (23-33 mg, 1.0 eq.), Cdc20 ligand 19a (24-35 mg, 1.1 eq.), HATU (24-30 mg, 1.2 eq.), and DIEA (18-25 mg, 3.0 eq.) were dissolved in DMF (2 mL), and the reaction system was stirred overnight at room temperature. TLC analysis showed that the reaction proceeds were completely reacted. The reaction solution was purified by Prep-HPLC to obtain BRD degrading agent MJC122-130, a white solid, at a yield of 33-55%, at 13-23 mg.

[0533]

[0534] Scheme 15Synthesis of PROTCs IVa(MJC631-637,n=1-7)

[0535] AR ligand AR-1 (40 mg, 1.0 eq.), linkers L3 of varying lengths (14-20 mg, 1.1 eq.), KI (2 mg, 0.2 eq.), and DIEA (34-50 mg, 3.0 eq.) were dissolved in DMF (2 mL). The reaction mixture was stirred overnight at 90 °C. TLC analysis showed the reaction was complete. The mixture was diluted with water and extracted with ethyl acetate, then dried over anhydrous sodium sulfate and concentrated. The crude product was dissolved in THF (2 mL), and saturated LiOH solution (0.5 mL) was added dropwise. The mixture was stirred at room temperature for 5 hours to remove the methyl ester. After the reaction was complete, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to obtain intermediate AR-L3 with linkers of varying lengths, 18-30 mg of white solid, with a two-step yield of 37-60%.

[0536] Intermediates AR-L3 (18-30 mg, 1.0 eq.), Apcin-A (14-20 mg, 1.1 eq.), HATU (17-24 mg, 1.2 eq.), and DIEA (18-25 mg, 3.0 eq.) were dissolved in DMF (2 mL), and the reaction mixture was stirred overnight at room temperature. TLC analysis showed that the reaction proceeds were completely reacted. The reaction solution was purified by Prep-HPLC to obtain AR degradation agent MJC631-637, a white solid, in a yield of 30-55% (8-25 mg).

[0537]

[0538] Scheme 16Synthesis of PROTCs IVb(MJC102-112,n=1-11)

[0539] JQ-acid (35 mg, 1.0 eq.), linkers of different lengths L4 (12-25 mg, 1.1 eq.), HATU (35 mg, 1.2 eq.), and DIEA (34 mg, 3.0 eq.) were dissolved in DMF (2 mL), and the reaction mixture was stirred overnight at room temperature. TLC analysis showed that the reaction proceeded completely. The mixture was diluted with water and extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated. The crude product was dissolved in THF (2 mL), and saturated LiOH solution (0.5 mL) was added dropwise. The mixture was stirred at room temperature for 5 hours to remove the methyl ester. After the reaction was complete, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to obtain intermediate JQ-L4 with linkers of different lengths, 20-35 mg of white solid, with a two-step yield of 40-62%.

[0540] Intermediates JQ-L4 (23-33 mg, 1.0 eq.), Apcin-A (14-20 mg, 1.1 eq.), HATU (17-24 mg, 1.2 eq.), and DIEA (18-25 mg, 3.0 eq.) were dissolved in DMF (2 mL), and the reaction mixture was stirred overnight at room temperature. TLC analysis showed that the reaction proceeds were completely reacted. The reaction solution was purified by Prep-HPLC to obtain the BET degrader MJC102-112, a white solid, at a yield of 39-59%, at a concentration of 15-26 mg.

[0541]

[0542] Scheme 17Synthesis of PROTCs IVb(MJC114-115,n=1-2)

[0543] JQ-acid (35 mg, 1.0 eq.), linkers of different lengths L5 (17-18 mg, 1.1 eq.), HATU (35 mg, 1.2 eq.), and DIEA (34 mg, 3.0 eq.) were dissolved in DMF (2 mL), and the reaction mixture was stirred overnight at room temperature. TLC analysis showed that the reaction proceeded completely. The mixture was diluted with water and extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated. The crude product was dissolved in THF (2 mL), and saturated LiOH solution (0.5 mL) was added dropwise. The mixture was stirred at room temperature for 5 h to remove the methyl ester. After the reaction was complete, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to obtain intermediate JQ-L5 with linkers of different lengths, 20-23 mg of white solid, with a two-step yield of 46-50%.

[0544] Intermediates JQ-L4 (20-23 mg, 1.0 eq.), Apcin-A (14-17 mg, 1.1 eq.), HATU (16-20 mg, 1.2 eq.), and DIEA (16-21 mg, 3.0 eq.) were dissolved in DMF (2 mL), and the reaction mixture was stirred overnight at room temperature. TLC analysis showed that the reaction proceeds were complete. The reaction solution was purified by Prep-HPLC to obtain the BET degrader MJC114-115, a white solid, at a yield of 15-20 mg (47-63%).

[0545] As exemplary portions of IIIa, IIIb, or combinations of IVa and IVb, some embodiments of this application involve portions of the Cdc20-based PROTACs structure as shown in Table 18:

[0546] Table 18. Some PROTACs structures based on Cdc20

[0547]

[0548]

[0549]

[0550]

[0551]

[0552]

[0553]

[0554]

[0555]

[0556]

[0557]

[0558]

[0559]

[0560]

[0561]

[0562]

[0563]

[0564]

[0565]

[0566]

[0567] Table 19 Activity tests of some PROTACs based on Cdc20

[0568]

[0569]

[0570]

[0571]

[0572]

[0573]

[0574]

[0575]

[0576]

[0577]

[0578]

[0579] A:1-10μM; B:10-50μM; C:50-100μM; D:>100μM

[0580] Example 14 Evaluation of the effect of reducing androgen receptor (AR) expression

[0581] In this experiment, the human prostate cancer cell line LNCaP was cultured in RPMI-1640 medium containing 10% fetal bovine serum (Gibco, USA) at 37°C, 5% CO2, and 95% humidity. When the cell density reached 80%, the cells were seeded into 6-well plates at 5 × 10⁶ cells per well. 5 Cells were cultured for 24 hours, and the culture medium containing the compound was changed to a final compound concentration of 0.01, 0.1, 1, and 10 μM. After 24 hours of treatment, the supernatant was removed, and the cells were washed twice with PBS. RIPA lysis buffer containing protease inhibitors and phosphatase inhibitors was added to the cells. After lysis and centrifugation, the total protein extract was obtained, and the protein concentration in the extract was detected by BCA method. Protein electrophoresis was performed by SDS-PAGE, and then the protein was transferred to a PVDF membrane by constant current electrophoresis at 200 mA for 90 min. The PVDF membrane was placed in 5% skim milk and blocked at room temperature for 1 h. Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273) (1:10000) was added, and the membrane was incubated overnight at 4 °C. The membrane was washed 3 times with TBST for 10 min each time. The membrane was incubated with secondary antibody (1:5000, absin) at room temperature for 60 min. The membrane was washed 3 times with TBST for 10 min each time. ECL luminescent solution was added and the membrane was exposed. Each sample was simultaneously detected using α-tubulin protein as an internal control. Protein maps are shown below. Figure 3 and Figure 4 The protein degradation rates are shown in Tables 20 and 21.

[0582] Table 20. Protein degradation rate of compounds

[0583]

[0584]

[0585] Table 21 Protein degradation rate at different concentrations

[0586] Compound numbering AR protein degradation rate (0.1 μM) AR protein degradation rate (1 μM) AR protein degradation rate (10 μM) MJC631 10％-20％ 40％-50％ 70％-80％ MJC632 10％-20％ 40％-50％ 70％-80％ MJC633 10％-20％ 40％-50％ 80％-90％ MJC634 20％-30％ 50％-60％ 80％-90％ MJC635 10％-20％ 40％-50％ 80％-90％ MJC636 40％-50％ 60％-70％ 90％-100％ MJC637 40％-50％ 50％-60％ 80％-90％

[0587] Example 15: Inhibitory activity of MJC636 on prostate cancer cell proliferation

[0588] Cell proliferation after compound treatment was assessed using the WST-1 assay. LNCaP (DMEM + 10% FBS) and 22Rv1 (MEM + 10% FBS) cells were passaged and seeded into 96-well cell culture plates at 8 × 10⁸ cells per well. 3After 24 hours, different concentrations of MJC636 were added to the cells, and the cells were incubated at 37°C in a 5% CO2 incubator for 4-7 days. Following the kit instructions, the absorbance of each well at 450 nm was measured using a microplate reader (zeroed with a blank control). Two replicates were set for each group. Statistical analysis and graphing were performed using GraphPad Prism 8 software. Figure 5 As shown in Table 22, the prostate cancer cell proliferation inhibitory activity of compound MJC636 is shown in Table 22.

[0589] Table 22 Inhibitory activity of compound MJC636 against prostate cancer cell proliferation

[0590] Compound numbering <![CDATA[LNCaP(IC 50 )]]> <![CDATA[22Rv1(IC 50 )]]> MJC636 3-4μM 4-5μM

[0591] Example 16 Evaluation of the effect of reducing the expression level of bromodomain protein (BRD)

[0592] In this experiment, human chronic myeloid monocytic leukemia MV-4-11 cells were cultured in IMEM medium containing 10% fetal bovine serum (Gibco, USA) at 37°C, 5% CO2, and 95% humidity. When the cell density reached 80%, a medium containing the compound was used to seed the cells into 6-well plates at a density of 1 x 10⁻⁶ cells / well. 6 / ml, to achieve final compound concentrations of 0.1, 1, 10, and 20 μM. After 24 hours of treatment, cells were collected, washed twice with PBS, and RIPA lysis buffer containing protease inhibitors and phosphatase inhibitors was added to the cells. After lysis and centrifugation, total protein extract was obtained, and the protein concentration in the extract was determined by the BCA method.

[0593] Protein electrophoresis was performed using SDS-PAGE, followed by transfer of proteins onto a PVDF membrane using a constant current of 200 mA for 90 min. The PVDF membrane was then blocked in 5% skim milk at room temperature for 1 h. BRD4 (#13440, CST) (1:1000) and C-MYC (ab32072, 1:1000) were added, and the membrane was incubated overnight at 4°C. The membrane was washed three times with TBST for 10 min each time. It was then incubated with secondary antibody (1:5000, absin) at room temperature for 60 min, washed three times with TBST for 10 min each time, and finally exposed to ECL chemiluminescence solution. GAPDH protein was also detected simultaneously in each sample as an internal control. Results are as follows: Figure 6 As shown in Table 7; the degradation effects are shown in Tables 23 and 24.

[0594] Table 23 Degradation of BRD4 protein by different compounds

[0595]

[0596]

[0597] Table 24 BRD4 protein degradation rate

[0598] Compound numbering BRD4 protein degradation rate (10 μM) MJC102 20％-30％ MJC103 20％-30％ MJC104 10％-20％ MJC105 0-10％ MJC106 0-10％ MJC107 0-10％ MJC108 50％-60％ MJC109 40％-50％ MJC110 50％-60％

[0599] Example 17 Inhibitory activity of MJC108 against cancer cell proliferation

[0600] Cell proliferation after compound treatment was assessed using the WST-1 assay. Human myeloid monocytic leukemia cells MV4-11 (DMEM + 10% FBS), human prostate cancer cells PC-3 (MEM + 10% FBS), and human breast cancer cells MCF-7 (MEM + 10% FBS) were passaged and seeded into 96-well cell culture plates at 8 × 10⁸ cells per well. 3 Cells were collected, and after 24 hours, different concentrations of MJC636 were added to the cells. The cells were then incubated at 37°C in a 5% CO2 incubator for 4-7 days. Following the kit instructions, the absorbance of each well at 450 nm was measured using a microplate reader (with a blank control set to zero). Two replicates were set for each group. Statistical analysis and graphing were performed using GraphPad Prism 8 software. Results are shown below. Figure 8 The cancer cell proliferation inhibitory activity of MJC108 is shown in Table 25.

[0601] Table 25 Inhibitory activity of MJC108 against cancer cell proliferation

[0602] Compound numbering <![CDATA[MV4-11(IC 50 )]]> <![CDATA[PC-3(IC 50 )]]> <![CDATA[MCF-7(IC 50 )]]> MJC108 1-2μM 2.49μM 4.43μM

Claims

1. A compound that binds to cyclin 20, said compound being as shown in Formula I; In structural formula I: X1, X2, and X3 may have one or two nitrogen atoms and the rest may be carbon atoms, or all of them may be carbon atoms. R1, R2, R3, R4, and R5 are simultaneously or partially -H, -OH, -OCH3, F, Cl, Br, -CF3, -OCF3, -CN, -NH2, -NO2, or C1-C6 straight-chain, branched, or cycloalkyl groups; Y1, Y2, Y3, and Y4 are all carbon atoms; R6, R7, R8, and R9 are simultaneously or partially -H, -CH3, -OCH3, -OCH2CH3, F, Cl, Br, or -CF3; X4, X5, and X6 may have one nitrogen atom and the rest may be carbon atoms, or all of them may be carbon atoms. When Y5, Y6, Y7, and Y8 are all carbon atoms, its structure is: , The corresponding substituent R 10 R 11 R 12 R 13 Simultaneously or partially -H or -CH3; When any one of the substituents in Y5, Y6, Y7, and Y8 is C, N, -CO-, O, S, or is absent, its corresponding substituent R 10 R 11 R 12 R 13 Simultaneously or partially -H or -CH3, X4, X5, and X6 are all carbon atoms, and their structure is as follows: , , , or ; R 14 Selected from -H, -OH, -OCH3, -OCH2CH3, -OCH2CH2CH3, F, Cl, Br, -CH2F, -CHF2, -CF3, -OCF3, -CN, -NH2, -NO2, or C1-C6 straight-chain, branched, cycloalkyl, phenyl; R 15 Selected from -NH2; W1 and W2 are -CONH-, -NHSO2-, or -SO2NH-; n1 and n2 are carbon chains of length 1, 2, 3, 4 or 5, and the configuration of chiral carbon 1 is R-type.

2. A compound that binds to cyclin 20, said compound having a structure selected from any of the following: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 , , , , , , , , , or .

3. A compound that binds to cyclin 20, the compound having the following structural formula: 。 4. A pharmaceutically acceptable salt of the compound of claim 1 or claim 2 that binds to cell division cycle protein 20, wherein the pharmaceutically acceptable salt is a salt formed by the preparation of a non-toxic acid or base addition of the compound of claim 1 or claim 2.

5. A protein degrading agent, said protein degrading agent being composed of a target protein ligand + linker + Cdc20 protein conjugate; wherein, The Cdc20 protein conjugate is the compound that binds cell cycle protein 20 as described in claim 1 or claim 2; the protein degrader is expressed as shown in (II); The target protein ligand is BRD. ; The Linker is selected from Linker 1 or Linker 2, and its structure is as follows: Linker 1 Or Linker 2 ; In Linker 1, A1 and B1 are independently selected from one or more of -CO- and -NH-, and m1 is selected from an integer from 0 to 20; in Linker 2, A2 and B2 are independently selected from one or more of -CO- and -NH-, and m2 is selected from an integer from 0 to 20.

6. A protein degrading agent, wherein the structure of the protein degrading agent is selected from any one of the following: , , , , , , , , , , , , , , , , , , , , , , or .

7. A protein degrading agent, wherein the structure of the protein degrading agent is selected from any one of the following: , , , , , , , , , , , , , , , or .

8. A pharmaceutical composition comprising the compound of claim 1 or claim 2 that binds to cyclin 20 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable diluent or carrier.

9. A pharmaceutical composition for targeting the degradation of a protein, said pharmaceutical composition comprising the protein degrading agent of claim 5 or claim 7 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable diluent or carrier.

10. The use of a compound of claim 1 or claim 2 that binds to cyclin 20, or a protein degrader of claim 5 or claim 7, in the preparation of a medicament for treating or preventing diseases caused by abnormal cell division promoted after Cdc20 activation of the APC / C complex; said disease being prostate cancer or breast cancer.

11. The use of a compound that binds to cyclin 20 in the preparation of a medicament for treating or preventing a disease caused by abnormal cell division promoted after Cdc20 activation of the APC / C complex; said disease being myeloid monocytic leukemia; the compound having the following structural formula: 。 12. The use of the compound of claim 1 or claim 2 that binds to cell division cycle protein 20, or the protein degrader of claim 5 or claim 7, in the preparation of an inhibitor of the Cdc20-activated APC / C complex.

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