Novel protein degradation agent conjugate and conjugate thereof

Abstract

The invention provides a novel protein degradation agent conjugate and a conjugate thereof. Specifically, the invention provides a protein binding conjugate or a pharmaceutically acceptable salt thereof, and the protein binding conjugate is as shown in a formula (I), wherein the variables are as defined herein, as well as pharmaceutical compositions containing the protein binding conjugates and uses thereof.

Description

Technical Field

[0001] This application belongs to the pharmaceutical field, specifically involving a new type of protein degradation agent conjugate and its coupling compounds. Background Technology

[0002] Epigenetic regulation is an effective strategy for treating diseases including cancer and fibrosis. Among these, BET family proteins play a crucial role in normal cell growth and cell cycle progression. Downstream genes of BET family member BRD4, such as MYC, CDK, and BCL2, play vital roles in the occurrence and development of diseases.

[0003] Protein degraders are an emerging drug intervention that can control diseases by degrading disease-related target proteins.

[0004] Although BET family protein-related inhibitors and protein degraders have shown some efficacy in preclinical and clinical trials, their effects are short-lived and require frequent dosing. Related antibody-protein degrader conjugates have also been reported, but their antitumor activity is poor due to the loss of activity from the released degrader, and the antibody portion requires engineered modification to achieve a high drug-to-antibody ratio (DAR).

[0005] In summary, there is an urgent need in this field to develop novel drug intervention methods targeting BET family proteins. Summary of the Invention

[0006] The purpose of this invention is to provide a novel class of protein degrading agents and their conjugates, and to provide conjugates comprising said conjugates. These degrading agents, conjugates, and conjugates can be used to treat cancer and fibrotic diseases.

[0007] In a first aspect of the invention, a protein binding conjugate or a pharmaceutically acceptable salt thereof is provided, said protein binding conjugate being shown in formula (I);

[0008]

[0009] in,

[0010] a is a value between 1 and 8 (inclusive of integers and non-inclusives);

[0011] L 1 For the connector part;

[0012] L P It can be a cleavable or incuttable linking group;

[0013] L 2 It is a branched group with or without one or more PEG units and / or polysarcosine units;

[0014] A is R E3 -R Linker -; where R E3 For the E3 ligase ligand moiety, R Linker For use with or without the E3 ligase ligand moiety and the BET family protein binding moiety as shown in Formula IA;

[0015]

[0016] L 3 Selected from the following group: amide, carbonyl, C1-8 alkylene amide, C1-8 alkylene carbonyl and C1-15 alkylene;

[0017] Ar is an aromatic ring or a heteroaromatic ring;

[0018] Bm is the binding portion that can specifically bind to the target protein.

[0019] In another preferred embodiment, a is a value between 2 and 8, more preferably, a is a value between 4 and 8, and even more preferably, a is a value between 6 and 8.

[0020] In another preferred embodiment, the BET family protein binding portion represented by formula IA is as shown in formula IA-1 or IA-2.

[0021]

[0022] In another preferred embodiment, the connector is a connector having a connecting group formed by a coupling reaction with the binding portion.

[0023] In another preferred embodiment, L 1 It is a protein binding linker.

[0024] In another preferred embodiment, L 1 -M Bm -L 1a -; where M Bm For the linking group formed by coupling reactions (such as lysine coupling, cysteine ​​coupling, genetically engineered non-natural amino acid coupling, and N-glycosylation coupling) with the binding moiety; L 1a It is a chain-like linker group.

[0025] In another preferred embodiment, L 1a It is a chain linking group with a length of 2 to 40 (preferably, 3 to 30, more preferably, 3 to 15, most preferably, 3 to 7) chain atoms.

[0026] In another preferred embodiment, the linking group (M) Bm Selected from the following group:

[0027] and This is the connection point with Bm.

[0028] In another preferred embodiment, L 1 -M Bm -L 1a -;in,

[0029] M Bm Selected from the following group: This is the connection point with Bm;

[0030] L 1a It is a chain-like linker group.

[0031] In another preferred embodiment, the protein binding conjugate is as shown in formula (I-1);

[0032]

[0033] Among them, M Bm and L 1a As defined above; L P L 2 L 3 A, Ar, Bm and a are defined as in equation (I).

[0034] In another preferred embodiment, L 1 -M Bm -M 1b -(M 1a ) p -M LP -;in,

[0035] M Bm As defined above;

[0036] M 1a Selected from the following group: C1-3 alkylene (preferably -CH2-), C1-2 alkylene -O (preferably -CH2CH2O-);

[0037] M 1b and M LP Each is independently selected from the group consisting of: none, C1-6 alkylene, C1-4 alkylene-CONH, C1-4 alkylene-CONH-C1-4 alkylene, C1-4 alkylene-NHCO, C1-2 alkylene-NHCO-C1-2 alkylene, C1-4 alkylene-CO, C1-4 alkylene-CO-C1-4 alkylene, C1-4 alkylene-NH, and C1-2 alkylene-NH-C1-2 alkylene; and

[0038] p is an integer from 0 to 10 (preferably, an integer from 1 to 10, more preferably, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10).

[0039] In another preferred embodiment, L 1a -M 1b -(M 1a ) p -M LP -

[0040] In another preferred embodiment, M LP To be with L P The connection position.

[0041] In another preferred embodiment, M 1a Selected from the following groups: -CH2-, -CH2CH2O-.

[0042] In another preferred embodiment, L 2 It is a branched group containing one or more PEG units and / or polysarcosine units, and M 1a When it is a C1-3 alkylene (preferably -CH2-), p is 3, 4, 5, 6 or 7.

[0043] In another preferred embodiment, L 2 It is a branched group containing one or more PEG units and / or polysarcosine units, and M 1a When it is a C1-2 alkylene-O (preferably -CH2CH2O-), p is 1 or 2.

[0044] In another preferred embodiment, L 2 When there is no time, M 1a It is a C1-2 alkylene-O (preferably -CH2CH2O-), with p being 5, 6, 7, 8, 9, or 10.

[0045] In another preferred embodiment, M LP Selected from the following group: none, -CO-, C1-2 alkylene, -C1-2 alkylene-CO-.

[0046] In another preferred embodiment, M LP It is -CO- or -C1-2 alkylene-CO-.

[0047] In another preferred embodiment, M 1b Selected from the group consisting of: none, C1-2 alkylene, -C≡C-, -C≡C-C1-2 alkylene-, -C1-2 alkylene-CONH-. In another preferred embodiment, M 1b It is either none or -C≡C-.

[0048] In another preferred embodiment, L 1 Selected from Table A1 below;

[0049] Table A1

[0050]

[0051] Where p is an integer from 1 to 10; For L P The connection point; This is the connection point for Bm.

[0052] In another preferred embodiment, L P It is a cleavable linker, preferably a protease-cleavable linker.

[0053] In another preferred embodiment, L P It is a linker group containing a polypeptide fragment.

[0054] In another preferred embodiment, the polypeptide fragment is a polypeptide fragment composed of 1, 2, 3, 4, 5 or 6 amino acid residues.

[0055] In another preferred embodiment, the amino acid residue is a residue of an amino acid selected from the group consisting of: citrulline, glutamine, glutamic acid, aspartic acid, alanine, glycine, valine, phenylalanine, D-phenylalanine, asparagine, and lysine.

[0056] In another preferred embodiment, the amino acid comprises an amino acid with an L- or D-configuration.

[0057] In another preferred embodiment, the amino acids include natural or non-natural amino acids (preferably, natural amino acids).

[0058] In another preferred embodiment, L P -M P2 -M P1 -M P3 -; where M P1 It is a polypeptide fragment; M P2 and M P3 Each independently is either non-NHCO-C(R) a )R b -CO-; where R a and R b Each is independently H or C1-4 alkyl; or R a and R b Together with the carbon atoms attached to them, they form C4-6 cycloalkyl groups.

[0059] In another preferred embodiment, M P2 and M P3 None.

[0060] In another preferred embodiment, M P2 and M P3 One is absent, the other is -NHCO-C(R)a )R b -CO-(Preferably, ).

[0061] In another preferred embodiment, L P for Among them, the better, To be with L 1 The connection point; For the aniline moiety (i.e. Connection point of NH); Z 1 and Z 2 Each is independently either absent (not present) or an amino acid residue, Z 3 and Z 4 Each is an amino acid residue.

[0062] In another preferred embodiment, Z 1 For the absence of glycine residues; and / or, Z 2 The following amino acid residues are absent or selected: L-glutamine residue, D-glutamine residue, L-glutamate residue, D-glutamate residue, L-aspartic acid residue, D-aspartic acid residue, L-alanine residue, D-alanine residue, and glycine residue; and / or, Z 3 Selected from the group consisting of: L-valine residues, D-valine residues, L-alanine residues, D-alanine residues, L-phenylalanine residues, D-phenylalanine residues, and glycine residues; and / or, Z 4 Selected from the following group: L-alanine residue, D-alanine residue, L-citrulline residue, D-citrulline residue, L-asparagine residue, D-asparagine residue, L-lysine residue, D-lysine residue, L-phenylalanine residue, D-phenylalanine residue, and glycine residue.

[0063] In another preferred embodiment, L P Selected from Table A2 below

[0064] Table A2

[0065]

[0066] in, To be with L 1 The connection point; and This is the connection point with the aniline portion.

[0067] In another preferred embodiment, L P As shown in equation (2.1) in Table A2.

[0068] In another preferred embodiment, the PEG unit is -(CH2CH2O)-, and / or the polysarcosine unit is -[CH2N(Me)CO]-.

[0069] In another preferred embodiment, L 2 In this context, the group containing one or more PEG unit structures and / or polysarcosine unit structures is -M. Ph -M 2L -M 2a -(CH2CH2O) q -M 2b or -M Ph -M 2L -M 2a -[CH2N(Me)CO] q -M 2b ;in,

[0070] M Ph Selected from the following group: none, C1-6 alkylene, 1 to 6 heteroalkylene (e.g., C1-2 alkylene-O-C1-2 alkylene);

[0071] M 2L Selected from the following group: none, -NHCO-, -N(C1-4 alkyl)CO-, linking groups formed by click chemistry (e.g. );

[0072] M 2a It is either non-alkyl or C1-4 alkylene;

[0073] M 2b Selected from the group consisting of: none, C1-2 alkyl, C1-2 alkylene-acid groups; and

[0074] q is an integer between 1 and 50 (preferably an integer between 10 and 40; more preferably an integer between 15 and 25).

[0075] In another preferred embodiment, M Ph It is a C1-6 alkylene or a 1 to 6 heteroalkylene (such as a C1-2 alkylene-O-C1-2 alkylene).

[0076] In another preferred embodiment, L 2 It is a group containing one or more PEG unit structures and / or polysarcosine unit structures.

[0077] In another preferred embodiment, M Ph For the aniline moiety (i.e. The connection position of the phenyl group.

[0078] In another preferred embodiment, the acidic group is an acidic group derived from an organic acid or an inorganic acid; preferably, the acidic group is selected from the group consisting of: carboxyl (-COOH), sulfonic acid (-SO3H), and phosphate (-OP(OH)3); more preferably, the acidic group is a carboxyl (-COOH).

[0079] In another preferred embodiment, L 2 Selected from Table A3 below

[0080] Table A3

[0081]

[0082] Where q is an integer between 1 and 50 (preferably an integer between 10 and 30), and q' is an integer between 1 and 6.

[0083] In another preferred embodiment, q' is 1, 2, or 3; preferably 1.

[0084] In another preferred embodiment, for

[0085] In another preferred embodiment, for

[0086] In another preferred embodiment, for

[0087] In another preferred embodiment, L 2 Connected to the aniline moiety (i.e. The meta position of the amino group or the ortho position of the methylene group in the )

[0088] In another preferred embodiment, L 3 In this context, the amide is -CONH-, -CON(C1-8 alkyl)-, and / or the carbonyl group is -CO- (i.e., L-). 3 In this context, the amide is -CONH- or -CON(C1-8 alkyl)-, the C1-8 alkylene amide is C1-8 alkylene--CONH- or C1-8 alkylene--CON(C1-8 alkyl)-, the carbonyl group is -CO-, and the C1-8 alkylene carbonyl group is C1-8 alkylene-CO-).

[0089] In another preferred embodiment, L 3 In the amide, the amino group is connected to A.

[0090] In another preferred embodiment, L 3 Selected from the following groups: -NHCO-, -(CH2) m - and -CO-; where m is an integer from 1 to 12.

[0091] In another preferred embodiment, R Linker This is a linker that is either absent or used to connect the E3 ligase ligand portion to the BET family protein binding portion as shown in Formula IA.

[0092] In another preferred embodiment, R Linker For none or as shown in the general formula selected from Table A4.1a;

[0093] Table A4.1a

[0094]

[0095] in, To and The connection point of the methylene group; n is an integer from 0 to 10 (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10); W is absent, carbonyl, C1-3 alkylene (e.g., methylene), C1-3 alkylene-carbonyl (preferably the carbonyl portion of the C1-3 alkylene-carbonyl group is connected to R). E3 (Connection); W L It is free of, NH or N (C1-3 alkyl); R L1 and R L2 Each is independently selected from the following group: H, C1-3 alkyl, C3-6 cycloalkyl, or R. L1 and R L2 And together with the carbon atoms connected to them, they form C3-6 cycloalkyl groups; W R It is a C3-10 cycloalkylene group or a 4- to 10-membered heteroalkylene group.

[0096] In another preferred embodiment, in equation (4.1.7a), n is 1 and / or W. L For none

[0097] In another preferred embodiment, R L1 and R L2 At most one of them is a C3-6 cycloalkyl group.

[0098] In another preferred embodiment, in equation (4.1.8a), W R It is a 4- to 10-membered heterocyclic alkyl group; preferably, the heterocyclic alkyl group contains only a nitrogen heteroatom as a heterocyclic atom, and more preferably, the nitrogen heteroatom is a linking position to other parts of the group. In another preferred embodiment, W L and W and W R The connection positions are spaced 1, 2 or 3 (preferably 2 or 3) ring atoms apart.

[0099] In another preferred embodiment, R i Linker As shown in Table A4.1a, formulas (4.1.1a), (4.1.2a), (4.1.3a), (4.1.5a), or (4.1.6a).

[0100] In another preferred embodiment, W is a carbonyl group or a methylene group.

[0101] In another preferred embodiment, in Table A4.1a, -CO- or W represents R E3 The connection position.

[0102] In another preferred embodiment, R E3 As shown in the general formula selected from Table A4.2a;

[0103] Table A4.2a

[0104]

[0105]

[0106] in,

[0107] For the aniline moiety (e.g.) The linker site of the methylene group;

[0108] X is either N or CH;

[0109] Ring B is an optionally substituted C3-20 cycloalkyl or an optionally substituted 4- to 20-membered heterocyclic alkyl;

[0110] The optional substitution refers to a group that is unsubstituted or one or more H atoms in the group being substituted by R atoms;

[0111] R is selected from the following group: hydroxyl, amino, C1-4 alkyl.

[0112] In another preferred embodiment, A is as shown in the general formula selected from Table A5a below;

[0113] Table A5a

[0114]

[0115] in,

[0116] To be with L 3 The connection point;

[0117] For the aniline moiety (e.g.) The linker site of the methylene group;

[0118] X is either N or CH;

[0119] n is an integer from 0 to 10 (such as 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10);

[0120] W is a carbonyl group or a C1-3 alkylene group (such as methylene); and

[0121] Cycle B is an optionally substituted C3-C20 cycloalkyl or an optionally substituted 4- to 20-membered heterocyclic alkyl;

[0122] The optional substitution refers to a group that is unsubstituted or one or more H atoms in the group being substituted by R atoms;

[0123] R is selected from the following group: hydroxyl, amino, C1-4 alkyl.

[0124] In another preferred embodiment, the cycloalkyl and heterocycloalkyl groups include: monocyclic, bridged, spirocyclic, or fused rings.

[0125] In another preferred embodiment, ring B is shown as in equation IB-1, IB-2, or IB-3:

[0126]

[0127] in,

[0128] X 1 X 2 X 3 and X 4 Each independently as X a X a -X b or X a -X b -X c

[0129] X 5 and X 6 Each independently is C(R) 1 ) or N;

[0130] X 7 None (single key), X a or X a -X b ;

[0131] X a X b and X c Each is independently selected from the following group: C(R) 1 )2、N(R 1 ), O and S; and

[0132] R 1 Each can be H or R independently.

[0133] In another preferred example, X a X b and X c Each is independently selected from the following group: C(R) 1 2. O and S.

[0134] In another preferred example, X a and X c For C(R) 1 )2,Xb For C(R) 1 2. O or S.

[0135] In another preferred embodiment, R 1 For H.

[0136] In another preferred example, X 1 X 2 X 3 and X 4 Each can be independently CH2, CH2-CH2, CH2-O, or CH2-CH2-CH2.

[0137] In another preferred embodiment, ring B is a ring as shown in formula IA-1 or IA-2.

[0138] In another preferred embodiment, ring B is a ring as shown in formula IA-1.

[0139] In another preferred embodiment, ring B is a ring as shown in equation IA-1; wherein, X 5 and X 6 Let N, X 1 and X 2 It is CH2.

[0140] In another preferred embodiment, Ar is a C6-10 aromatic ring or a 5- to 10-membered heteroaromatic ring (preferably, the heteroaromatic ring is a nitrogen-containing heteroaromatic ring, more preferably, the nitrogen-containing heteroaromatic ring contains only nitrogen atoms as heteroatoms); preferably, Ar is a phenyl or a 5- or 6-membered nitrogen-containing heteroaromatic ring (such as pyridine). In another preferred embodiment, the nitrogen-containing heteroaromatic ring contains 1, 2, or 3 nitrogen atoms as heterocyclic atoms, and the remainder are carbocyclic atoms.

[0141] In another preferred embodiment, the binding portion is selected from the group consisting of: an antibody, an antibody fragment of the antibody, or an antigen-binding fragment of the antibody.

[0142] In another preferred embodiment, Bm is an antibody or its antigen-binding portion.

[0143] In another preferred embodiment, the target protein is a surface antigen.

[0144] In another preferred embodiment, the surface antigens include 5T4, ACE, ADRB3, AKAP-4, ALK, androgen receptor, AOC3, APP, axonin 1, AXL, B7H3, B7-H4, BCL2, BCMA, bcr-abl, BORIS, BST2, C242, C4.4a, and CA. 125, CA6, CA9, CAIX, CCL11, CCR5, CD123, CD133, CD138, CD142, CD15, CD15-3, CD171, CD179a, CD18, CD19, CD19-9, CD2, CD20, CD22, CD23, CD24, CD25, CD27L, CD28, CD3, CD30, CD31, CD300LF, CD33, CD352, CD37, CD38, CD4, CD40, CD41, CD44, CD44v6, CD5 CD51, CD52, CD54, CD56, CD62E, CD62P, CD62L, CD70, CD71, CD72, CD74, CD79a, CD79b, CD80, CD90, CD97, CD125, CD138, CD141, CD147, CD152, CD154, CD326, CEA, CEACAM5, CFTR, condensation factor, cKit, closure protein 3, closure protein 18.2, CLDN6, CLEC12A, CLL-1, cll3, c-MET, Crypto 1. Growth factors, CS1, CTLA-4, CXCR2, CXORF61, cyclin B1, CYP1B1, cadherin-3, cadherin-6, DLL3, E7, EDNRB, EFNA4, EGFR, EGFRvIII, ELF2M, EMR2, ENPP3, EPCAM, EphA2, liver glycoside A4, liver glycoside B2, EPHB4, ERBB2 (Her2 / neu), ErbB3, ERG (TMPRSS2) ETS fusion gene), ETBR, ETV6-AML, FAP, FCAR, FCRL5, FGFR1, FGFR2, FGFR3, FGFR4, FLT3, folate receptor α, folate receptor β, FOLR1, Fos-associated antigen 1, fucose GM1, GCC, GD2, GD3, GloboH, GM3, GPC1, GPC2, GPC3, gplOO, GPNMB, GPR20, GPRC5D, GUCY2C, HAVCR1, HER2, HER3, HGF, HMI.24. HMWMAA, HPV E6, hTERT, human telomerase reverse transcriptase, ICAM, ICOS-L, IFN-α, IFN-γ, IGF-I receptor, IGLL1, IL-2 receptor, IL-4 receptor, IL-13Ra2, IL-11Ra, IL-1, IL-12, IL-23, IL-13, IL-22, IL-4, IL-5, IL-6, interferon receptor, integrins (including α4, α). v β3, α v β5, α v β6, α1β4, α4β1, α4β7, α5β1, α6β4, α IIbβ3 integrin), integrin αV, intestinal carboxyl esterase, KIT, LAGE-1a, LAIR1, LAMP-1, LCK, podin, LewisY, LFA-1 (CD11a), L-selectin (CD62L), LILRA2, LIV-1, LMP2, LRRC15, LY6E, LY6K, LY75, MAD-CT-1, MAD-CT-2, MAGE A1, MelanA / MART1, mesothelin, MLIAP, MSLN, mucin, MUC1, MUC16, mut hsp70-2, MYCN, myostatin, NA17, NaPi2b, NCA-90, NCAM, connexin-4, NGF, NOTCH1, NOTCH2, NOTCH3, NOTCH4, NY-BR-1, NY-ESO-1, o-acetyl-GD2, OR51E2, OY-TES1, p53, p53 mutant, PANX3, PAP, PAX3, PAX5, p-CAD, PCTA-1 / galactagoguerin 8, PD-L1, PD-L2, PDGFR, PDGFR-β, phosphatidylserine, PIK3CA, PLAC1, polysialic acid, prostate enzymes, prostate cancer cells, prostaglandins, copper *Pseudomonas aeruginosa*, rabies virus, survivin and telomerase, PRSS21, PSCA, PSMA, PTK7, RAGE-1, RANKL, Ras mutant, respiratory syncytial virus, rhesus monkey factor, RhoC, RON, ROR1, ROR2, RU1, RU2, sarcoma translocation breakpoint, SART3, SLAMF7, SLC44A4, sLe, SLITRK6, spermin 17, sphingosine 1-phosphate, SSEA-4, SSX2, STEAP1, TAG72, TARP, TCRβ, TEM1 / CD248, TEM7R, tendinin C, TF, TGF-1, TGF-β2, TNF-α, TGS5, Tie 2. TIM-1, TnAg, TRAC, TRAIL-R1, TRAIL-R2, TROP-2, TRP-2, TRPV1, TSHR, tumor antigen CTAA16.88, tyrosinase, UPK2, VEGF, VEGFR1, VEGFR2, vimentin, WT1, XAGE1, or combinations thereof.

[0145] In another preferred embodiment, the surface antigen includes: HER2, TROP-2, B7H3, EGFR, CD3, CD19, CD20, CD22, CD30, CD33, CD37, CD38, CD52, CD79b, CD138, BCMA, CTLA-4, CA125, Nectin-4, TF, CEGF, folate receptor α, or combinations thereof.

[0146] In another preferred embodiment, Bm is an antibody or an antibody fragment or an antigen-binding fragment thereof as defined below.

[0147] In another preferred embodiment, the antibody is selected from the group consisting of: rituximab, trastuzumab, depatuxizumab, dicetuzumab, gemtuzumab, vobramitamab, pertuzumab, obbituzumab, oflamuzumab, olatozumab, ixartuzumab, denintuzumab, inelizumab, ifinatamab, moxetumomab, mirvetuximab, vebutuzumab, datopotamab, sacituzumab, u3-1784, daratumumab, STI-6129, lintuzumab, naltuzumab, oxintuzumab, belantan, polotuzumab, iladatuzumab, indatuzumab, cetuximab, anti-CD38. a2 antibody, alemtuzumab, teimozumab, belantuzumab, atorxizumab, indatuximab, faucizumab, enroximateb, texizumab, tosimomumab, bevacizumab, panitumumab, trimemumab, texizumab, caputsuzumab, ozovumab, and vetozumab.

[0148] In another preferred embodiment, the antibody is selected from the group consisting of: trastuzumab, pertuzumab, decituzumab, datopotamab, sacituzumab, vobramitamab, ifinatamab, depatuxizumab, cetuximab, and panitumumab.

[0149] In another preferred embodiment, the antibody is selected from the group consisting of trastuzumab, ifinatamab, cetuximab, and sacituzumab.

[0150] In another preferred embodiment, the protein binding conjugate is selected from Table B below;

[0151] Table B: Protein Conjugates

[0152]

[0153]

[0154]

[0155] In a second aspect of the invention, a pharmaceutical composition is provided comprising (i) a protein binding conjugate as described in the first aspect or a pharmaceutically acceptable salt thereof, and (ii) one or more pharmaceutically acceptable carriers.

[0156] In a third aspect of the invention, the use of a protein binding conjugate as described in the first aspect or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cancer is provided.

[0157] In another preferred embodiment, the cancer is selected from the group consisting of: breast cancer, lung cancer, gastric cancer, hepatocellular carcinoma, lymphoma, leukemia, pancreatic cancer, head and neck cancer, squamous cell carcinoma, urethral cancer, colorectal cancer, prostate cancer, ovarian cancer, bladder cancer, gastrointestinal stromal tumor, cervical cancer, esophageal cancer, peritoneal cancer, liver cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, vulvar cancer, thyroid cancer, penile cancer, plasmacytoma, myeloma, sarcoma, or combinations thereof.

[0158] In another preferred embodiment, the drug for treating cancer may be used in combination with an adjuvant.

[0159] In another preferred embodiment, the adjuvant is a cytotoxic agent or an immunomodulator.

[0160] In another preferred embodiment, the immune response modulator is a checkpoint inhibitor.

[0161] In another preferred embodiment, the checkpoint inhibitor includes a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a TIM3 inhibitor, and / or a LAG-3 inhibitor.

[0162] In a fourth aspect of the invention, a method for treating cancer is provided, the method comprising administering to a subject in need a therapeutically effective amount of a protein binding conjugate as described in the first aspect or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a protein binding conjugate as described in the first aspect or a pharmaceutically acceptable salt thereof.

[0163] In another preferred embodiment, the cancer is as defined above.

[0164] In another preferred embodiment, the method further includes administering a therapeutically effective amount of adjuvant to the subject.

[0165] In another preferred embodiment, the adjuvant may be administered to the subject before, simultaneously with, or after the protein binding conjugate as described in the first aspect or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the protein binding conjugate as described in the first aspect or a pharmaceutically acceptable salt thereof.

[0166] In another preferred embodiment, the additive is as defined above.

[0167] In a fifth aspect of the invention, a degrading agent or a pharmaceutically acceptable salt thereof is provided, said degrading agent being as shown in formula (II);

[0168]

[0169] in,

[0170] Ar and L 3 The definition is the same as the definition in formula (I);

[0171] A i For R i E3 -R i Linker -; where R i E3 For the E3 ligase ligand moiety, R i Linker It is either absent or used to connect the E3 ligase ligand moiety.

[0172] In another preferred embodiment, R i Linker For none or as shown in the general formula selected from Table A4.1b;

[0173] Table A4.1b

[0174]

[0175] Among them, R i N H or C1-4 alkyl; W L 、W R R L1 R L2 W and n are defined in the general formula shown in Table A4.1a.

[0176] In another preferred embodiment, n is an integer from 0 to 10 (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10); W is a carbonyl or C1-3 alkylene group (e.g., methylene).

[0177] In another preferred embodiment, R i Linker As shown in Table A4.1b, formulas (4.1.1b), (4.1.2b), (4.1.3b), (4.1.5b), or (4.1.6b).

[0178] In another preferred embodiment, R i E3 As shown in the general formula selected from Table A4.2b;

[0179] Table A4.2b

[0180]

[0181]

[0182] X and ring B are defined in the general formula shown in Table A4.2a.

[0183] In another preferred example, A i From Table A5b below

[0184] Table A5b

[0185]

[0186] in, To be with L 3 The connection points; the definitions of X, n, W and ring B are the same as those in the general formula shown in Table A5a.

[0187] In another preferred embodiment, the BET family protein binding site represented by formula IA is as shown in formula IA-1 or IA-2, R i Linker As shown in Table A4.1b, formulas (4.1.1b), (4.1.2b), (4.1.3b), (4.1.5b), or (4.1.6b), and R i E3 As shown in Table A4.2b, equations (4.2.7b), (4.2.8b), (4.2.9b), (4.2.13b), (4.2.14b), (4.2.15b), or (4.2.16b) (preferably, equations (4.2.7b), (4.2.8b), or (4.2.9b)).

[0188] In another preferred embodiment, the BET family protein binding portion represented by formula IA is as shown in IA-2 (preferably, in formula IA-2, L...). 3 It is a C1-2 alkylene group, more preferably, L 3 Selected from the following group: amide, carbonyl, C1-8 alkylene amide, C1-8 alkylene carbonyl and C1 alkylene (i.e., methylene), R i Linker As shown in Table A4.1b, equations (4.1.1b), (4.1.2b), (4.1.3b), (4.1.5b), or (4.1.6b) (preferably, equation (4.1.1b) or (4.1.3b)), and R i E3 As shown in Table A4.2b, equations (4.2.1b), (4.2.2b), (4.2.3b), (4.2.4b), (4.2.5b), (4.2.6b), (4.2.10b), (4.2.11b), (4.2.12b) (preferably, equations (4.2.1b), (4.2.2b), or (4.2.10b)) are shown.

[0189] In another preferred embodiment, the degrading agent is selected from Table C below;

[0190] Table C

[0191]

[0192]

[0193]

[0194]

[0195] In a sixth aspect of the invention, a pharmaceutical composition is provided, characterized in that it comprises (i) a degrading agent as described in the fifth aspect or a pharmaceutically acceptable salt thereof, and (ii) one or more pharmaceutically acceptable carriers.

[0196] In a seventh aspect of the invention, the use of a degrading agent as described in the fifth aspect or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cancer is provided.

[0197] In another preferred embodiment, the cancer is as defined above.

[0198] In another preferred embodiment, the drug for treating cancer may be used in combination with an adjuvant.

[0199] In another preferred embodiment, the additive is as defined above.

[0200] In an eighth aspect of the invention, a method for treating cancer is provided, the method comprising administering to a subject in need a therapeutically effective amount of a degrading agent as described in the first aspect or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a degrading agent as described in the first aspect or a pharmaceutically acceptable salt thereof.

[0201] In another preferred embodiment, the cancer is as defined above.

[0202] In another preferred embodiment, the method further includes administering a therapeutically effective amount of adjuvant to the subject.

[0203] In another preferred embodiment, the adjuvant may be administered to the subject before, simultaneously with, or after the protein binding conjugate as described in the first aspect or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the protein binding conjugate as described in the first aspect or a pharmaceutically acceptable salt thereof.

[0204] In another preferred embodiment, the additive is as defined above.

[0205] In a ninth aspect of the invention, a connector-degrader conjugate or a pharmaceutically acceptable salt thereof is provided, said connector-degrader conjugate being shown in formula (III);

[0206]

[0207] Among them, Ar, L3 A and L 2 As defined in equation (I); L ii Bm This is to enable further coupling of linkers that can specifically bind to the binding site of the target protein.

[0208] In another preferred embodiment, L ii Bm For M ii Bm -L 1a -L P ; among which, L P and L 1a As defined in equation (I).

[0209] In another preferred embodiment, the connector-degrader coupling is as shown in formula (III-1);

[0210]

[0211] Among them, Ar, L 3 A, L 2 and L P As defined in equation (I), L ii 1 This is to enable further coupling of the linker portion that can specifically bind to the target protein.

[0212] In another preferred embodiment, the L ii 1 or L ii Bm It has reactive groups (M) capable of coupling with binding sites (such as antibodies) (e.g., lysine coupling, cysteine ​​coupling, genetically engineered non-natural amino acid coupling, and N-glycosylation coupling). ii Bm ).

[0213] In another preferred embodiment, the connector-degrader coupling is as shown in formula (III-2);

[0214]

[0215] Among them, Ar, L 3 A, L 2 L P and L 1a As defined in equation (I), M ii Bm A reactive group that can undergo a coupling reaction with the binding moiety.

[0216] In another preferred embodiment, M ii Bm for

[0217] In another preferred embodiment, the connector-degrader conjugate is selected from Table D below.

[0218] Table D: Connector-Degrader Couplings

[0219]

[0220]

[0221]

[0222] In a tenth aspect of the present invention, a method for preparing a conjugate (antibody conjugate) is provided, the conjugate being as shown in Formula I-1; and the preparation method includes the steps of:

[0223] A mixture comprising a linker-degrader conjugate as described in the ninth aspect and a reduced antibody is provided; the reduced antibody is coupled to the linker-degrader conjugate to obtain a conjugate as shown in Formula I-1.

[0224] In a tenth aspect of the invention, the use of the linker-degrader conjugate as described in the ninth aspect in the preparation of the protein-binding conjugate as described in the first aspect or a pharmaceutically acceptable salt thereof is provided.

[0225] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0226] Figure 1 The SEC characterization spectra of representative conjugates of this application are shown.

[0227] Figure 2 The degradation activity of the representative novel degrading agent of this application is shown (immunoblotting).

[0228] Figure 3 The differences in degradation activity of the representative conjugates of this application in different cell lines were shown (immunoblotting). Detailed Implementation

[0229] Through long-term and in-depth research, the inventors have discovered a new class of BET degrading agents (as shown in Formula II) and conjugates containing these degrading agents (as shown in Formula I), both of which exhibit excellent antitumor activity. Furthermore, the inventors have also discovered that the new BET degrading agents (as shown in Formula II) can be modified with specific structures... The linker-degrader conjugate obtained from the linker fragment (as shown in Formula III) exhibits excellent antibody compatibility. High-purity and high-DAR conjugates can be prepared without special conjugation techniques or antibody modification, and the resulting conjugates show good stability (low aggregation) and demonstrate excellent and sustained antitumor effects at low dosing frequencies and doses. Based on this, the inventors completed this invention.

[0230] the term

[0231] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. As used herein, when referring to a specifically enumerated numerical value, the term “about” means that the value can vary from the enumerated value by no more than 1%. For example, the expression “about 100” includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0232] As used herein, unless otherwise defined, the term "alkyl" itself, or as part of another substituent, refers to a straight-chain or branched hydrocarbon group having a specified number of carbon atoms (i.e., C64 ... 1-6 (Indicates 1-6 carbons). Preferably, the alkyl group specifically has 1-4 carbons, i.e., C1-C2. 1-4 Alkyl groups. Examples of alkyl groups include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, etc.

[0233] As used herein, unless otherwise stated, the term "heteroalkyl" itself or as part of another substituent refers to a group formed by the substitution of one or more carbon atoms in an alkyl group with a specified number of carbon atoms as defined above, by one to three heteroatoms selected from O, N, and S. The total number of carbon atoms and heteroatoms in a heteroalkyl group is expressed as a subset (e.g., 3-4-membered heteroalkyl) in which nitrogen and sulfur atoms may optionally be oxidized, and nitrogen heteroatoms may optionally be quaternized. Heteroatoms O, N, and S may be located at any internal position of the heteroalkyl group. Examples include -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2, -S(O)-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=N-OCH3, and -CH=CH-N(CH3)-CH3. At most two heteroatoms can be consecutive, for example, -CH2-NH-OCH3 and -CH2-O-Si(CH3)3.

[0234] As used herein, the term "alkylene" itself, or as part of another substituent, refers to a divalent group derived from an alkane as defined above. Alkylenes typically have 1-24 (preferably 1-10) carbon atoms.

[0235] As used herein, the term “heteroalkylene” itself or as part of another substituent refers to a saturated or unsaturated or polyunsaturated divalent group derived from a heteroalkylene group, such as -CH2-CH2-S-CH2CH2- and -CH2-O-CH2-CH2-CH2-.

[0236] As used herein, the term "cycloalkyl" refers to a hydrocarbon ring (preferably fully saturated) having a specified number of ring atoms (e.g., C3-20 cycloalkyl, C4-6 cycloalkyl) and having no more than one double bond between the ring apexes. This term also includes bicyclic and polycyclic hydrocarbon rings, such as bridged rings, fused rings, spirocyclic rings, etc. The term "heterocyclic alkyl" refers to a cycloalkyl ring having a specified number of ring atoms (e.g., 4-20 membered heterocyclic alkyl) and containing one to five heteroatoms selected from N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom is optionally quaternized. Heterocyclic alkyl can be monocyclic, bicyclic, or polycyclic systems (e.g., bridged rings, fused rings, spirocyclic rings, etc.). Non-limiting examples of heterocyclic alkyl groups include pyrrolidine, imidazoline, pyrazolidine, butyrolactam, valproic acid, imidazolidinone, hydantoin, dioxolane, benzodiimide, piperidine, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrrolidine, thiaran, pyranone, tetrahydrofuran, tetrahydrothiophene, quinine ring, etc. Heterocyclic alkyl groups can be attached to the rest of the molecule via a cyclic carbon or heteroatom.

[0237] Unless otherwise stated, the term "aryl" refers to a polyunsaturated (usually aromatic) hydrocarbon group, which can be monocyclic or fused together or covalently linked polycyclic (up to three rings). The term "heteroaryl" refers to an aryl group (or ring) containing 1 to 5 heteroatoms selected from N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom is optionally quaternized. Heteroaryl groups can be attached to the rest of the molecule via carbon atoms or heteroatoms. Non-limiting examples of aryl groups include phenyl, naphthyl, and biphenyl, while non-limiting examples of heteroaryl groups include pyridyl, pyrazinyl, pyrazinyl, pyrimidinyl, triazinyl, quinolinyl, quinoxalinyl, quinazolinyl, terpineyl, phthalazinyl, benzotriazinyl, purine, benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzoisoxazolyl, and isobenzofuranyl. furyl), isoindolyl, indoleyl, benzotriazinyl, thienopyridyl, thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridine, benzothiazolyl, benzofuranyl, benzothiaphenyl, indolyl, quinolinyl, isoquinolinyl, isothiazolyl, pyrazolyl, indazoleyl, pteridinyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrroleyl, thiazolyl, furanyl, thiaphenyl, etc.

[0238] In some embodiments, the terms above (such as "alkyl", "aryl", and "heteroaryl") will include substituted and unsubstituted forms of the specified group. Preferred substituents for each type of group are provided below. For brevity, the terms aryl and heteroaryl will refer to the substituted or unsubstituted forms as provided below, while the term "alkyl" and the associated aliphatic group refer to the unsubstituted form unless specified otherwise.

[0239] The substituents of alkyl groups (including those commonly referred to as alkylene, alkenyl, ynyl, and cycloalkyl) can be a variety of groups selected from the group consisting of: -halogen, -OR', -NR'R”, -SR', -SiR'R”R”', -OC(O)R', -C(O)R', -CO2R', -CONR'R”, -OC(O)NR'R”, -NR”C(O)R', -NR'-C(O)NR”R”', -NR” C(O)₂R', -NH-C(NH₂)=NH, -NR'C(NH₂)=NH, -NH-C(NH₂)=NR', -S(O)R', -S(O)₂R', -S(O)₂NR'R”, -NR'S(O)₂R”, -CN, and -NO₂, in quantities ranging from zero to (2M'+1), where M' is the total number of carbon atoms in this group. R', R”, and R”’ each independently represent hydrogen, and unsubstituted C 1-8 Alkyl, unsubstituted heteroalkyl, unsubstituted aryl, aryl substituted with 1-3 halogens, unsubstituted C 1-8 Alkyl, C 1-8 Alkoxy or C1-8 Thioalkoxy, or unsubstituted aryl-C 1-4 Alkyl groups. When R' and R” are attached to the same nitrogen atom, they can combine with the nitrogen atom to form 3-, 4-, 5-, 6-, or 7-membered rings. For example, -NR'R” refers to a group comprising 1-pyrrolidinyl and 4-morpholinyl. The term “acyl”, used alone or as part of another group, refers to a group in which the substituents on the carbon closest to the connection point of the group are substituted with =O (e.g., -C(O)CH3, -C(O)CH2CH2OR', etc.).

[0240] Similarly, the substituents of aryl and heteroaryl groups are diverse and are usually selected from: -halogen, -OR', -OC(O)R', -NR'R”, -SR', -R', -CN, -NO2, -CO2R', -CONR'R”, -C(O)R', -OC(O)NR'R”, -NR”C(O)R', -NR”C(O)2R', -NR'-C(O)NR”R”', -NH- C(NH2)=NH, -NR'C(NH2)=NH, -NH-C(NH2)=NR', -S(O)R', -S(O)2R', -S(O)2NR'R”, -NR'S(O)2R”, -N3, perfluoro(C1-C4)alkoxy and perfluoro(C1-C4)alkyl, in numbers ranging from zero to the total number of open valences on the aromatic ring system; wherein R', R” and R”' are independently selected from hydrogen, C 1-8 Alkyl, C 3-6 cycloalkyl, C 2-8 alkenyl, C 2-8 Alkynyl, unsubstituted aryl and heteroaryl, (unsubstituted aryl)-C 1-4 Alkyl and unsubstituted aryloxy-C 1-4 Alkyl groups. Other suitable substituents include each of the above aryl substituents attached to the ring atom via an alkylene chain of 1-4 carbon atoms.

[0241] As used herein, the term "amino acid residue" refers to the group formed by removing an H atom from the N-terminal -NH2 group and the -OH atom from the C-terminal -COOH group of an amino acid. Generally, the segment of an amino acid (residue) including both the N-terminus and the C-terminus is called the main chain, while the portion that determines the specific type of amino acid is called the side chain. Unless otherwise defined, in this document, amino acids include both native and non-native amino acids, including D-type and / or L-type amino acids. Examples of amino acids include, but are not limited to, Ala (A), Arg I, Asn (N), Asp (D), Cys (C), Gln (Q), GI (E), Gly (G), His (H), Ile (I), Leu (L), Lys (K), Met (M), Phe (F), Pro (P), Ser (S), Thr (T), Trp (W), Tyr (Y), and Val (V). Preferably, in this document, the amino acid is selected from the group consisting of: L-glycine (L-Gly), L-alanine (L-Ala), β-alanine (β-Ala), L-glutamic acid (L-Glu), L-aspartic acid (L-Asp), L-histidine (L-His), L-arginine (L-Arg), L-lysine (L-Lys), L-valine (L-Val), L-serine (L-Ser), and L-threonine (L-Thr); furthermore, when the amino acid has two or more amino groups and / or two or more carboxyl groups, the term also includes groups formed by removing one H from -NH2 and -OH from -COOH on different carbon atoms, such as the divalent group -C(O)-(CH2)2-C(COOH)-NH- formed by removing one H from -NH2 and non-α-COOH of glutamic acid respectively.

[0242] In this document, unless otherwise defined, bonds indicated by dashed lines or marked with wavy lines represent positions that connect to other parts of the molecule.

[0243] For the compounds presented herein, the bond from the substituent (usually an R group) to the center of the ring will be understood as a bond that provides a connection at any available vertex of the aromatic ring.

[0244] In this invention, the term "pharmaceuticalally acceptable" refers to a substance that is suitable for use in humans and / or animals without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), i.e., a substance with a reasonable benefit / risk ratio.

[0245] In this invention, the term "effective amount" or "therapeutic effective amount" refers to the amount of a therapeutic agent that treats, alleviates, or prevents a target disease or condition, or the amount that exhibits a detectable therapeutic or preventative effect. The precise effective amount for a given subject depends on that subject's body size and health condition, the nature and severity of the condition, and the choice of the therapeutic agent and / or combination of therapeutic agents administered. Therefore, it is useless to pre-specify an accurate effective amount. However, for a given condition, the effective amount can be determined using routine experiments, and a clinician can judge it accordingly.

[0246] Unless otherwise specified, all compounds mentioned in this invention are intended to include all possible optical isomers, such as compounds with a single chirality, or mixtures of various chiral compounds (i.e., racemates). In all compounds of this invention, each chiral carbon atom may optionally be in the R configuration or the S configuration, or a mixture of the R and S configurations.

[0247] As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by the compounds of the present invention with an acid or base that is suitable for use as a medicine. Pharmaceutically acceptable salts include both inorganic and organic salts. A preferred class of salts are those formed by the compounds of the present invention with an acid. Suitable acids for forming salts include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenesulfonic acid, and benzenesulfonic acid; and acidic amino acids such as aspartic acid and glutamic acid.

[0248] Unless otherwise specified, the term “amino acid” as used in this article is intended to include any common amino acid, such as aspartic acid, glutamic acid, cysteine, asparagine, phenylalanine, glutamine, tyrosine, serine, methionine, tryptophan, glycine, valine, leucine, alanine, isoleucine, proline, threonine, histidine, lysine, and arginine.

[0249] Unless otherwise defined, each structural formula (such as...) The range of subscripts (such as m, n, p, q, q') in brackets, represented by ab, refers to the range including the endpoint values, i.e., a and b, and all integers in between. For example, n = 12-24 means that n can be 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24, and m = 1-6 means that m can be 1, 2, 3, 4, 5, or 6.

[0250] The protein-binding conjugates, compositions containing them, and applications of the present invention.

[0251] As used herein, the term "protein-binding conjugate" (also referred to herein as "the conjugate of the present invention") refers to a class of conjugates having a binding moiety, as shown in Formula (I), attached to which a specific protein (such as a target protein) is attached. When the binding moiety is an antibody or a fragment thereof or an antigen-antigen binding fragment thereof, "the conjugate of the present invention" may also be referred to as "the antibody conjugate of the present invention".

[0252] In some respects, this article provides a protein binding conjugate or a pharmaceutically acceptable salt thereof, wherein the protein binding conjugate is as shown in formula (I);

[0253]

[0254] Among them, a and L 1 L P L 2 A, L 3 Ar and Bm are as defined in the first aspect.

[0255] In some implementations, a is a value between 1 and 8; L 1 For the connector part; L P It is a polypeptide fragment; L 2 A represents the branched portion; A represents the E3 ligase ligand and linker; L represents the branched portion. 3 It is an amide, carbonyl, or alkyl chain; Ar is an aromatic ring or heterocyclic ring; and Bm is a binding moiety that can specifically bind to proteins.

[0256] In this document, the "drug / antibody ratio" (DAR) refers to the ratio of the degrading agent to the binding moiety, and in Formula I, 'a' can represent the selectable range of this ratio. For the protein-binding conjugates described herein, generally, high DAR or relatively high DAR means a ratio of 4-8, preferably 6-8; low DAR or relatively low DAR means a ratio less than 4.

[0257] In some implementations, the binding portion is an antibody, an antibody fragment, or an antigen-binding fragment.

[0258] In some implementations, L 1 It is a protein binding linker.

[0259] In some implementations, L1 is selected from Table A1 described above.

[0260] In some implementations, L P Selected from Table A mentioned above or from the following table:

[0261]

[0262] Z1, Z2, Z3 and Z4 each independently do not contain or are L- or D-configured naturally occurring amino acid residues, provided that at least two of Z1, Z2, Z3 and Z4 are amino acid residues; This is the connection point with L1; This is the connection point for the aniline portion on the right.

[0263] In some implementation schemes, Z 1 It may be absent or contain glycine; Z 2 The group consisting of or not present in the group containing: L-glutamine, D-glutamine, L-glutamic acid, D-glutamic acid, L-aspartic acid, D-aspartic acid, L-alanine, D-alanine, and glycine; Z 3 Choose from the following groups: L-valine, D-valine, L-alanine, D-alanine, L-phenylalanine, D-phenylalanine, and glycine; and Z. 4 Choose from the following groups: L-alanine, D-alanine, L-citrulline, D-citrulline, L-asparagine, D-asparagine, L-lysine, D-lysine, L-phenylalanine, D-phenylalanine, and glycine.

[0264] In another implementation, L 2 It does not exist or is selected from Table A3 mentioned above.

[0265] In some implementations, L 3 Selected from the following group: and Where m is an integer from 1 to 12.

[0266] In some implementations, A is selected from Table A5a described above.

[0267] In some implementations, in Table A5a, X is N or CH; n is an integer from 0 to 10; W is carbonyl or methylene; B is an aliphatic heterocycle or aliphatic spiroheterocycle; To be with L 3 The connection point; and This is the connection point with the left benzyl portion.

[0268] In some implementations, Ar is a substituted or unsubstituted aromatic ring or heterocyclic ring.

[0269] In some embodiments, Bm is an antibody or its antigen-binding moiety. In some aspects, the protein bound to the binding moiety is a surface antigen. In some embodiments, the surface antigen and antibody are as defined above.

[0270] In some implementations, the protein binding conjugate is selected from Table B above.

[0271] In some implementation schemes, a, L 1 L P L 2 A, L 3 Ar and Bm are each independently the corresponding groups in the specific conjugates shown in Table B or the specific compounds prepared in the examples (such as the conjugates of the degradation agent conjugates prepared in Examples 1-3). In some embodiments, Bm, L 1 、M Bm L 1a L P 、M P2、 M P3、 R a R b 、M P1 L 2 、M Ph 、M 2L 、M 2a 、M 2b q, A, R E3 X, ring B, X1 to X7, X a X b X c , R, R Linker , W, n, L 3 m, Ar, and a are each independently the corresponding groups in the specific conjugates shown in Table B or the specific compounds prepared in the examples (such as the conjugates of the degradation agent conjugates prepared in Example 3).

[0272] In some respects, this article also provides a pharmaceutical composition comprising the said conjugate or compound or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

[0273] In some aspects, this document also provides a method for treating cancer in a subject of need, the method comprising administering to the subject a pharmaceutically acceptable or therapeutically effective amount of the conjugate or a composition containing thereof, or a pharmaceutically acceptable salt thereof. In some embodiments, the cancer is breast cancer, lung cancer, gastric cancer, hepatocellular carcinoma, lymphoma, leukemia, pancreatic cancer, head and neck cancer, squamous cell carcinoma, urethral cancer, colorectal cancer, prostate cancer, ovarian cancer, bladder cancer, gastrointestinal stromal tumor, cervical cancer, esophageal cancer, peritoneal cancer, liver cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, vulvar cancer, thyroid cancer, penile cancer, plasmacytoma, myeloma, or sarcoma.

[0274] In some embodiments, the method includes administering a pharmaceutically acceptable or therapeutically effective amount of an adjuvant to the subject before, after, or simultaneously with the conjugate or composition or a pharmaceutically acceptable salt thereof. In some embodiments, the adjuvant is a cytotoxic agent or an immune response modulator. In some embodiments, the immune response modulator is a checkpoint inhibitor. In some embodiments, the checkpoint inhibitor includes PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, TIM3 inhibitors, and / or LAG-3 inhibitors.

[0275] The degradation agent, compositions containing the degradation agent, and applications of the present invention.

[0276] As used in this article, a degrading agent refers to a class of compounds that can target and induce protein degradation via the ubiquitination-proteasome pathway. Generally, such degrading agents are bifunctional small molecules formed by linking the ligand of the target protein (such as BET family proteins) to an E3 ubiquitin ligase ligand, for example, through a linker. They can simultaneously bind to both the target protein and the E3 ubiquitin ligase, causing the target protein to approach the E3 ubiquitin ligase and be ubiquitinated, thereby being recognized and degraded by the intracellular proteasome.

[0277] In some respects, this article provides novel degrading agents or pharmaceutically acceptable salts thereof, said degrading agents as shown in formula (II);

[0278]

[0279] Among them, A i As defined in the fifth aspect; Ar and L 3 As defined in Equation I.

[0280] In some implementations, Ar is an aromatic ring or a heterocyclic aromatic ring.

[0281] In some implementation schemes, A i For E3 ligase ligands and linkers.

[0282] In some implementation schemes, A i Selected from Table A5b mentioned above.

[0283] In some implementations, Table A5b, To be with L 3 For some connection points, X is N or CH; and / or n is an integer from 0 to 10; and / or W is carbonyl or methylene; and / or B is an aliphatic heterocycle or aliphatic spirocyclic heterocycle.

[0284] In some implementations, L 3 It can be an amide, carbonyl, or alkyl chain.

[0285] In some implementations, L 3 Selected from the following group: Where m is an integer from 1 to 12.

[0286] In some implementation schemes, A i L 3 Ar and Ar are each independently the corresponding groups in the specific degrading agents shown in Table C or the specific compounds prepared in the examples (such as the degrading agents prepared in Example 1). In some embodiments, A i R i E3 X, ring B, X1 to X7, X a X b X c , R, R i Linker , W, n, L 3 Each of m and Ar is an independent group in the specific degrading agent shown in Table C or the specific compound prepared in the examples (such as the degrading agent prepared in Example 1).

[0287] In some implementations, the degrading agent is selected from Table C above.

[0288] In some respects, this document also provides a conjugate formed by linking the degrading agent to a binding moiety (Bm) capable of specifically binding to a target protein via a linker as shown in Formula IIA;

[0289]

[0290] Among them, L 1 L P L 2 As defined above, M D It is a linker group formed after reacting with the hydroxyl or amino group (such as -NH-) in the linker or E3 ligase ligand in the degradation agent.

[0291] In some implementation schemes, M D This refers to the linker group formed after reacting with an amino group (such as -NH-) in the linker of the degradation agent or a hydroxyl group in the E3 ligase ligand moiety. In some embodiments, M... D It is either absent or -CO-O-. In some embodiments, the linker group formed with the amino group (such as -NH-) in the linker of the degrading agent is -CO-O-, while the linker group formed after reacting with the hydroxyl group in the E3 ligase ligand moiety is absent.

[0292] In some respects, this article also provides a pharmaceutical composition comprising the aforementioned degrading agent or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

[0293] In some aspects, this document also provides a method for treating cancer in a subject of need, the method comprising administering to the subject a pharmaceutically acceptable or therapeutically effective amount of the degradation agent or a composition containing the agent, or a pharmaceutically acceptable salt thereof. In some embodiments, the cancer is as previously defined.

[0294] In some embodiments, the method includes administering a pharmaceutically acceptable or therapeutically effective amount of an adjuvant to the subject before, after, or simultaneously with the degrading agent or composition or a pharmaceutically acceptable salt thereof. In some embodiments, the adjuvant is as defined above.

[0295] The connector-degrader coupling of the present invention and its application

[0296] As used herein, the term “connector-degrader conjugate” (also referred to herein as “degrader conjugate”) refers to a class of conjugates that, as in formula (III), can be further linked to a binding moiety that can specifically bind to a protein (such as a target protein).

[0297] In some aspects, this document also provides a connector-degrader coupling, wherein the connector-degrader coupling is as shown in formula (III), formula (III-1) and formula (III-2);

[0298]

[0299] Among them, L ii Bm L ii 1 and M ii Bm As defined in the ninth aspect, L 1a L P L 2 L 3 A and Ar are defined as in equation (I).

[0300] In some embodiments, the linker-degrader conjugate can be used to conjugate with binding moieties (such as antibodies, their antibody fragments, or their antigen-binding fragments) capable of specifically binding to target proteins.

[0301] In some implementations, L ii Bm L ii 1 、M ii Bm L 1a L P L 2 L 3A and Ar are each independently the corresponding groups in the specific connector-degrader conjugate shown in Table D or the specific compound prepared in the examples (such as the degrader conjugate prepared in Example 2). In some embodiments, L ii Bm L ii 1 、M ii Bm L 1a L P 、M P2 、M P3 R a R b 、M P1 L 2 、M Ph 、M 2L 、M 2a 、M 2b q, A, R E3 X, ring B, X1 to X7, X a X b X c , R, R Linker , W, n, L 3 m, Ar, and a are each independently the corresponding groups in the specific connector-degrader conjugate shown in Table D or the specific compound prepared in the examples (such as the degrader conjugate prepared in Example 2).

[0302] In some embodiments, the connector-degrader conjugate is selected from Table D above.

[0303] In some aspects, this document also provides a method for preparing the conjugate (or antibody conjugate) of the present invention using the aforementioned linker-degrader conjugate, comprising the steps of: providing a mixture containing the aforementioned linker-degrader conjugate and a reduced antibody; and conjugating the reduced antibody with the linker-degrader conjugate to obtain the conjugate.

[0304] Pharmaceutical Compositions and Administration

[0305] Because the protein-binding conjugates (sometimes also called conjugates or antibody-drug conjugates) or degrading agents provided by this invention have excellent ability to degrade BET family proteins and antitumor activity, the conjugates (or antibody-drug conjugates) or degrading agents (especially conjugates) of this invention can be used to treat a variety of cancers (such as breast cancer, lung cancer, gastric cancer, hepatocellular carcinoma, lymphoma, leukemia, pancreatic cancer, head and neck cancer, squamous cell carcinoma, urethral cancer, colorectal cancer, prostate cancer, ovarian cancer, bladder cancer, gastrointestinal stromal tumors, cervical cancer, esophageal cancer, peritoneal cancer, liver cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, vulvar cancer, thyroid cancer, penile cancer, plasmacytoma, myeloma, or sarcoma, etc.).

[0306] The aforementioned protein-binding conjugates or degrading agents can be administered to subjects (e.g., humans) in therapeutically effective amounts via appropriate routes. For conjugate drugs (or antibody-drug conjugates), subjects requiring treatment may be patients at risk or suspected of having a condition related to the activity or expression level of a specific antigen. Such patients can be identified through routine physical examinations.

[0307] Conventional methods, known to those skilled in the art of medicine, can be used to administer the pharmaceutical composition to a subject, depending on the type of disease to be treated or the site of the disease. This composition can also be administered via other conventional routes, such as parenteral administration, oral administration, via inhalation spray, topical administration, rectal administration, nasal administration, oral administration, vaginal administration, or via implantation. The term "parenteral" as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intra-articular, intrasynovial, intrasternal, intrathecal, intralesional, and intracranial injection or infusion techniques. Furthermore, it can be administered via injectable reservoir routes, such as using injectable or biodegradable materials and methods with 1-, 3-, or 6-month reservoirs.

[0308] Injectable compositions may contain various carriers such as vegetable oils, dimethylactamide, dimethylformamide, ethyl lactate, ethyl carbonate, isopropyl myristate, ethanol, polyols (glycerol, propylene glycol, liquid polyethylene glycol, etc.). For intravenous injection, water-soluble antibodies may be administered via infusion, thereby delivering a pharmaceutical formulation containing the antibody and physiologically acceptable excipients. Physiologically acceptable excipients may include, for example, 5% glucose, 0.9% saline, Ringer's solution, or other suitable excipients. Intramuscular preparations, such as sterile formulations of a suitable soluble salt form of the antibody, may dissolve and administer pharmaceutical excipients such as water-based injections, 0.9% saline, or 5% glucose solutions.

[0309] When treated with the protein-binding conjugates or degrading agents of the present invention, delivery can be performed using methods conventional in the art. For example, it can be introduced into cells using liposomes, hydrogels, cyclodextrins, biodegradable nanocapsules, or bioadhesive microspheres. Alternatively, the nucleic acid or carrier can be delivered locally by direct injection or by using an infusion pump. Other methods include various transport and carrier systems using conjugates and biodegradable polymers.

[0310] The pharmaceutical compositions of the present invention contain a safe and effective amount of the protein-binding conjugate or degrading agent of the present invention and a pharmaceutically acceptable carrier. Such carriers include (but are not limited to): saline, buffer solutions, glucose, water, glycerol, ethanol, and combinations thereof. Generally, the pharmaceutical formulation should be matched to the route of administration. The pharmaceutical compositions of the present invention can be formulated as solutions, for example, prepared using conventional methods with physiological saline or aqueous solutions containing glucose and other excipients. The pharmaceutical compositions are preferably manufactured under aseptic conditions. The dosage of the active ingredient is a therapeutically effective amount.

[0311] The effective amount of the protein-binding conjugate (or antibody-drug conjugate) or degrader described in this invention can vary depending on the administration method and the severity of the disease to be treated. A preferred effective amount can be determined by those skilled in the art based on various factors (e.g., through clinical trials). These factors include, but are not limited to: pharmacokinetic parameters of the antibody-drug conjugate, such as bioavailability, metabolism, and half-life; the severity of the disease to be treated, the patient's weight, the patient's immune status, and the route of administration. Generally, satisfactory results are obtained when the antibody-drug conjugate of this invention is administered at a dose of approximately 0.01 mg to 50 mg / kg animal body weight (preferably 0.1 mg to 20 mg / kg animal body weight). For example, due to the urgency of the treatment condition, several separate doses may be administered monthly, or the dose may be reduced proportionally.

[0312] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalers. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be necessary.

[0313] The compounds of this invention can be administered alone or in combination with other pharmaceutically acceptable therapeutic agents or adjuvants.

[0314] When using the pharmaceutical composition, a safe and effective amount of the antibody-drug conjugate of the present invention is applied to the mammal (such as a human) requiring treatment. The dosage administered is the pharmaceutically considered effective dose. For a person weighing 60 kg, the single dose is typically 1–2000 mg, preferably 5–500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the scope of the skills of a skilled physician.

[0315] The main advantages of this invention include:

[0316] Compared to existing technologies, the novel protein degrader described in this invention exhibits superior antitumor activity. Its derived conjugates are highly compatible with antibodies and can be used to prepare high-purity and high-DAR conjugates with antibodies such as trastuzumab, cetuximab, sacituzumab, and ifinatamab without the need for site-directed conjugation technology. Furthermore, it demonstrates excellent and sustained antitumor effects in in vivo animal models at low dosing frequencies.

[0317] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight.

[0318] Example 1. Synthesis of a new degrading agent

[0319] Option 1. Synthesis of key intermediates for new degradation agents

[0320]

[0321] Step 1: Synthesis of Compound 2. Compound 1 (20 g, 102.47 mmol) and N-bromosuccinimide (27.36 g, 153.71 mmol) were dissolved in carbon tetrachloride (160 mL), and azobisisobutyronitrile (1.01 g, 6.15 mmol) was added. The reaction was carried out overnight at 75 °C under nitrogen protection. After the reaction was complete as monitored by TLC, it was quenched with sodium thiosulfate solution, extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was dissolved in tetrahydrofuran, and diethyl phosphite (4 mL) and N,N-diisopropylethylamine (4 mL) were added. The reaction was carried out for 3 hours at room temperature. After the reaction was complete as monitored by TLC, it was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was directly used in the next step.

[0322] Step 2: Synthesis of Compound 3. Compound 2 (28.08 g, 102.46 mmol) and sodium methanesulfonate (26.15 g, 256.14 mmol) were added to acetonitrile (200 mL) and reacted at 65 °C for 4 hours. After the reaction was completed by TLC monitoring, the mixture was extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was slurried with petroleum ether to give the title compound 3 (18.0 g, yield 64%). 1 H NMR (400MHz, DMSO-d6) δ8.13(d,J=8.3Hz,1H),7.91(d,J=1.9Hz,1H),7.86-7.83(m,1H),4.75(s,2H),3.88(s,3H),3.00(s,3H).LC-MS(ESI):m / z 273.0[M+H] + .

[0323] Step 3: Synthesis of Compound 4. Compound 3 (18 g, 65.87 mmol), iron powder (18.39 g, 329.36 mmol), and ammonium chloride (17.64 g, 329.36 mmol) were added to methanol (240 mL) and water (60 mL) and refluxed at 65 °C for 2 hours. After the reaction was complete as monitored by TLC, the mixture was filtered. The filtrate was extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was slurried with petroleum ether to give the title compound 4 (14.4 g, 90% yield). 1 H NMR (400MHz, DMSO-d6) δ7.76 (d, J = 2.2Hz, 1H), 7.32-7.24 (m, 1H), 6.84-6.75 (m, 3H), 4.31 (s, 2H), 3.80 (s, 3H), 2.85 (s, 3H). LC-MS (ESI): m / z 243.1[M+H]+ .

[0324] Step 4: Synthesis of Compound 6. Compound 4 (6 g, 24.66 mmol), Compound 5 (4.18 mL, 36.99 mmol), palladium acetate (1.38 g, 6.17 mmol), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (5.88 g, 12.33 mmol), and cesium carbonate (22.5 g, 69.06 mmol) were added to 1,4-dioxane (200 mL) and reacted at 90 °C for 3 hours under nitrogen protection. After the reaction was complete as monitored by TLC, the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (ethyl acetate / dichloromethane = 0-80%) to give the title compound 6 (2.04 g, yield 23%). 1 H NMR (400MHz, DMSO-d6) δ9.22 (s, 1H), 7.97 (d, J = 2.2Hz, 1H), 7.57-7.49 (m, 1H), 7.45-7.38 (m, 2H ),7.18-7.11(m,1H),6.88-6.81(m,1H),4.43(s,2H),3.89(s,3H),2.90(s,3H).LC-MS(ESI):m / z 355.1[M+H] + .

[0325] Step 5: Synthesis of Compound 7. Compound 6 (2.04 g, 5.57 mmol) and N-bromosuccinimide (1.02 g, 5.57 mmol) were dissolved in trifluoroacetic acid (15 mL) and reacted at room temperature for 1 hour. After the reaction was completed by TLC monitoring, the mixture was concentrated under reduced pressure, and the residue was slurried with petroleum ether to give the title compound 7 (2.3 g, yield 92.08%). 1 H NMR(400MHz,DMSO-d6)δ8.19(s,1H),7.93-7.88(m,2H),7.29-7.21(m,1H),6.94-6.88(m,1H), 6.67(dt,J=9.3,4.6Hz,1H),4.57(s,2H),3.72(d,J=3.1Hz,3H),2.96(s,3H).LC-MS(ESI):m / z 435.0[M+H] + .

[0326] Step 6: Synthesis of Compound 9. Compound 8 (4 g, 10.49 mmol), pinacol diborate (5.6 g, 22.03 mmol), palladium dichloride (736.3 mg, 1.05 mmol), and potassium acetate (2.2 g, 22.03 mmol) were dissolved in dimethyltetrahydrofuran (40 mL) and reacted overnight at 80 °C under nitrogen protection. After the reaction was complete as monitored by TLC, the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (ethyl acetate / petroleum ether = 0-60%) to give the title compound 9 (2.55 g, yield 57%). 1 H NMR(400MHz,DMSO-d6)δ7.98(d,J=3.4Hz,1H),7.94-7.87(m,2H),7.73(s,1H),7.41(d,J=8 .2Hz,2H),6.82(d,J=3.4Hz,1H),3.43(s,3H),2.38(s,3H),1.29(s,12H).LC-MS(ESI):m / z 429.2[M+H] + .

[0327] Step 7: Synthesis of Compound 10. Compound 7 (2.30 g, 5.30 mmol), Compound 9 (2.84 g, 6.62 mmol), tetrakis(triphenylphosphine)palladium (1.53 g, 1.32 mmol), and potassium carbonate (1.83 g, 13.24 mmol) were added to 1,4-dioxane (100 mL) and water (25 mL). The reaction was carried out at 80 °C for 5 hours under nitrogen protection. After the reaction was completed by TLC monitoring, the mixture was extracted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (ethyl acetate / dichloromethane = 0%–80%) to give the title compound 10 (3.24 g, yield 86%). LC-MS (ESI): m / z 656.2 [M+H] + .

[0328] Step 8: Synthesis of Compound 11. Compound 10 (3.24 g, 4.94 mmol) and paraformaldehyde (445.1 mg, 14.82 mmol) were dissolved in trifluoroacetic acid (30 mL) and reacted at 45 °C for 20 min. After the reaction was completed by TLC monitoring, the mixture was concentrated under reduced pressure, and the residue was slurried with diethyl ether to give the title compound 11 (2.35 g, yield 71%). LC-MS (ESI): m / z 668.2 [M+H] + .

[0329] Step 9: Synthesis of Compound 12. Compound 11 (2.3 g, 3.44 mmol) and lithium hydroxide monohydrate (867.27 mg, 20.67 mmol) were dissolved in methanol (100 mL), tetrahydrofuran (20 mL), and water (30 mL), and reacted overnight at 40 °C. After the reaction was monitored by TLC until complete, the pH was adjusted to acidic, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the title compound 12 (1.62 g, 91% yield). 1 H NMR (600MHz, DMSO-d6) δ12.89(s,1H),11.85(d,J=2.8Hz,1H),7.99(d,J=2.1Hz,1H ),7.73(s,1H),7.52(d,J=2.1Hz,1H),7.21(d,J=2.6Hz,1H),6.94(ddd,J=12.2,8. 8,2.9Hz,1H),6.67(ddd,J=10.3,8.4,2.8Hz,1H),6.43(td,J=9.7,5.8Hz,1H),5.1 3(d,J=16.6Hz,1H),4.70-4.36(m,3H),3.61(s,3H),3.01(s,3H).LC-MS(ESI):m / z 500.1[M+H] + .

[0330] Step 10: Synthesis of Compound 13. Compound 12 (300 mg, 0.6 mmol) was added to a boranetetrahydrofuran complex (2 M, 5 mL) and reacted overnight at 50 °C. After the reaction was complete as monitored by LC-MS, it was quenched with methanol, concentrated under reduced pressure, and the residue was purified by column chromatography (methanol / dichloromethane = 0-5%) to give an intermediate. This intermediate (220 mg, 0.45 mmol) and Dys-Martin oxidant (384 mg, 0.91 mmol) were dissolved in dichloromethane (15 mL) and reacted at room temperature for 2 hours. After the reaction was complete as monitored by LC-MS, it was extracted with dichloromethane, the organic phase was washed once with 10% sodium thiosulfate aqueous solution and once with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (methanol / dichloromethane = 0-5%) to give the title compound 13 (210 mg, 96% yield). LC-MS(ESI): m / z 484.2 [M+H] + .

[0331] Step 11: Synthesis of Compound 14. Compound 13 (180 mg, 0.37 mmol) and (benzyloxycarbonylmethylene)triphenylphosphine (168 mg, 0.41 mmol) were dissolved in dichloromethane (10 mL) and reacted overnight at room temperature. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure, and the residue was purified by column chromatography (ethyl acetate / dichloromethane = 0-5%) to give the title compound 14 (200 mg, yield 88%). LC-MS (ESI): m / z 616.2 [M+H] + .

[0332] Step 12: Synthesis of Compound 15. Compound 14 (200 mg, 0.32 mmol) and 10% palladium on carbon (20 mg) were added to methanol (7 mL) and reacted overnight at room temperature under hydrogen atmosphere. After the reaction was monitored by LC-MS until complete, the mixture was filtered, and the filtrate was concentrated under reduced pressure to give the corresponding title compound 15 (160 mg, 95% yield). LC-MS (ESI): m / z 528.2 [M+H] + .

[0333] Step 13: Synthesis of Compound 17: Compound 16 (600 mg, 1.2 mmol) was dissolved in a boranetetrahydrofuran complex (2 M, 15 mL) and reacted overnight at 60 °C. After the reaction was complete as monitored by LC-MS, methanol was added to quench the reaction, and the mixture was concentrated under reduced pressure. The residue was purified by column chromatography (methanol / dichloromethane = 0-5%) to give the intermediate. This intermediate and Dysmart oxidant (1.53 g, 3.6 mmol) were dissolved in dichloromethane (30 mL) and reacted at room temperature for 2 hours. After the reaction was complete as monitored by LC-MS, the mixture was extracted with dichloromethane, and the organic phase was washed once with 10% sodium thiosulfate aqueous solution and once with saturated brine. The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (methanol / dichloromethane = 0-5%) to give the title compound 17 (542 mg, 93% yield). LC-MS (ESI): m / z 485.2 [M+H] + .

[0334] Option 2. Synthesis of new degradation agents XZ1701 series and XZ1706 series

[0335]

[0336] General Step 1: Compound 12 (1 equivalent), compound 18 or 19 (1.2 equivalent), and N,N-diisopropylethylamine (4 equivalent) were dissolved in dimethyl sulfoxide (2 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.5 equivalent) were added. The mixture was reacted at room temperature for 30 minutes. After the reaction was completed as monitored by LC-MS, the title compound was obtained by preparative liquid chromatography.

[0337] Step 1.1. Synthesis of compound XZ1701-C4. The title compound (11.2 mg) was obtained from compounds 12 and 18-C4 using a general procedure. 1 H NMR (600MHz, DMSO-d6) δ11.84(d,J=2.6Hz,1H),8.99(s,1H),8.38(d,J=7.8Hz,1H),8.16-8.04(m,1H),7.93(d,J=2.1Hz,1H),7.81(d,J=9.2Hz,1H), 7.74(s,1H),7.44(d,J=8.0Hz,2H),7.38(d,J=8.0Hz,2H),7.34(s,1H),7. 20(d,J=3.9Hz,1H),6.95-6.87(m,1H),6.65-6.59(m,1H),6.38-6.27(m,1H ),5.16-5.03(m,2H),4.97-4.88(m,1H),4.67-4.48(m,4H),4.48-4.41(m, 1H),4.29(s,1H),3.62(s,4H),3.18(s,2H),3.02(s,4H),2.46(s,3H),2.32 -2.20(m,1H),2.17-2.10(m,1H),2.05-1.99(m,1H),1.86-1.77(m,1H),1. 69-1.54(m,2H),1.38(d,J=7.0Hz,3H),1.00-0.89(m,9H).LC-MS(ESI):m / z 1011.4[M+H] + .

[0338] Step 1.2. Synthesis of compound XZ1701-C6. Using compounds 12 and 18-C6 as starting materials, the title compound (10.2 mg) was obtained using a general procedure. 1H NMR (600MHz, DMSO-d6) δ11.83(d,1H),8.99(s,1H),8.38(d,J=7.8Hz,1H),8.09-8.03(m,1H),7.93(d,J=2.1Hz,1H),7.78(d,J=9.5Hz,1H),7.73 (s,1H),7.44(d,J=8.2Hz,2H),7.39(d,J=8.2Hz,2H),7.34(d,J=2.1Hz,1H),7.21(d,J=2.6Hz,1H),6.96-6.89(m,1H),6.65-6.58(m,1H),6.35- 6.27(m,1H),5.15-5.03(m,2H),4.97-4.87(m,1H),4.66-4.39(m,5H),4 .29(s,1H),3.67-3.58(m,4H),3.17-3.10(m,1H),3.01(s,3H),2.55(s, 1H),2.46(s,2H),2.27-2.20(m,1H),2.16-2.08(m,1H),2.05-1.98(m,1 H),1.84-1.77(m,1H),1.67-1.15(m,11H),0.95(s,9H).LC-MS(ESI):m / z 1039.5[M+H] + .

[0339] Step 1.3. Synthesis of compound XZ1701-C8. The title compound (8.5 mg) was obtained from compounds 12 and 18-C8 using a general procedure. 1H NMR (600MHz, DMSO-d6) δ11.84(d,J=2.7Hz,1H),8.99(s,1H),8.37(d,J=7.8Hz ,1H),8.10-8.03(m,1H),7.93(d,J=2.2Hz,1H),7.79(d,J=9.3Hz,1H),7.73(s ,1H),7.44(d,J=7.9Hz,2H),7.38(d,J=7.9Hz,2H),7.34(d,J=2.0Hz,1H),7.2 1(d,J=2.7Hz,1H),6.96-6.90(m,1H),6.64-6.58(m,1H),6.36-6.24(m,1H),5. 12-5.03(m,2H),4.96-4.89(m,1H),4.62(d,J=13.6Hz,1H),4.58-4.48(m,3H) ,4.43(t,J=8.0Hz,1H),4.29(s,1H),3.61(s,3H),3.17-3.12(m,1H),3.01(s,4 H),2.55(s,1H),2.46(s,3H),2.29-2.23(m,1H),2.16-2.09(m,1H),2.05-1.9 8(m,1H),1.85-1.76(m,1H),1.53-1.17(m,14H),0.94(s,9H).LC-MS(ESI):m / z 1067.4 [M+H] + .

[0340] Step 1.4. Synthesis of compound XZ1701-C9. The title compound (18.0 mg) was obtained from compounds 12 and 18-C9 using a general procedure. 1H NMR(600MHz,DMSO-d6)δ11.83(d,J=2.7Hz,1H),8.98(s,1H),8.37(d,J=7.8Hz,1H), 8.05(t,J=5.7Hz,1H),7.92(d,J=2.1Hz,1H),7.78(d,J=9.3Hz,1H),7.72(s,1H),7. 45-7.42(m,2H),7.40-7.36(m,2H),7.33(d,J=2.0Hz,1H),7.20(d,J=2.6Hz,1H),6. 94-6.88(m,1H),6.63-6.58(m,1H),6.31-6.25(m,1H),5.11-5.04(m,2H),4.96-4.8 6(m,1H),4.62(d,J=13.6Hz,1H),4.56-4.49(m,3H),4.42(t,J=8.1Hz,1H),4.30-4. 25(m,1H),3.64-3.57(m,5H),3.16-3.10(m,1H),3.07-3.02(m,1H),3.01(s,3H),2. 45(s,3H),2.28-2.22(m,1H),2.14-2.07(m,1H),2.04-1.98(m,1H),1.82-1.76(m,1 H),1.54-1.42(m,2H),1.39-1.31(m,5H),1.22(s,8H),0.94(s,9H).LC-MS(ESI):m / z 1081.4[M+H] + .

[0341] Step 1.5. Synthesis of compound XZ1701-C10. Using compounds 12 and 18-C10 as starting materials, the title compound (12.0 mg) was obtained using a general procedure. 1H NMR(600MHz,DMSO-d6)δ11.83(s,1H),8.99(s,1H),8.42-8.32(m,1H),8.08- 8.01(m,1H),7.93(d,J=2.1Hz,1H),7.78(d,J=9.3Hz,1H),7.73(s,1H),7.44( d,J=7.9Hz,2H),7.38(d,J=8.0Hz,2H),7.34(d,J=2.0Hz,1H),7.20(d,J=2.5 Hz,1H),6.97-6.86(m,1H),6.66-6.56(m,1H),6.34-6.23(m,1H),5.11-5.04( m,2H),4.97-4.88(m,1H),4.62(d,J=13.5Hz,1H),4.57-4.48(m,3H),4.43(t ,J=8.0Hz,1H),4.31-4.26(m,1H),3.61(s,4H),3.13(s,1H),3.01(s,4H),2.4 6(s,3H),2.30-2.22(m,1H),2.16-2.08(m,1H),2.04-1.98(m,1H),1.83-1.76 (m,1H),1.56-1.31(m,8H),1.27-1.19(m,10H),0.94(s,9H).LC-MS(ESI):m / z 1095.5 [M+H] + .

[0342] Step 1.6. Synthesis of compound XZ1701-C11. Using compounds 12 and 18-C11 as starting materials, the title compound (9.0 mg) was obtained using a general procedure. 1H NMR(600MHz,DMSO-d6)δ11.83(d,J=2.8Hz,1H),8.98(s,1H),8.37(d,J=7.8Hz,1H), 8.05(t,J=5.7Hz,1H),7.92(d,J=2.1Hz,1H),7.78(d,J=9.3Hz,1H),7.72(s,1H),7. 45-7.36(m,4H),7.33(d,J=2.0Hz,1H),7.20(d,J=2.7Hz,1H),6.93-6.88(m,1H),6. 63-6.57(m,1H),6.28(td,J=9.7,5.8Hz,1H),5.10(d,J=3.5Hz,1H),5.06(d,J=16.6 Hz,1H),4.95-4.88(m,1H),4.62(d,J=13.5Hz,1H),4.56-4.46(m,3H),4.42(t,J=8. 0Hz,1H),4.28(s,1H),3.65-3.55(m,5H),3.18-3.08(m,1H),3.09-2.97(m,4H),2.4 5(s,3H),2.29-2.22(m,1H),2.14-2.06(m,1H),2.04-1.98(m,1H),1.82-1.76(m,1H ),1.55-1.41(m,2H),1.37-1.17(m,17H),0.93(s,9H).LC-MS(ESI):m / z1109.4[M+H] + .

[0343] Step 1.7. Synthesis of compound XZ1701-C12. Using compounds 12 and 18-C12 as starting materials, the title compound (18.0 mg) was obtained using a general procedure. 1H NMR (600MHz, DMSO-d6) δ11.83(s,1H),8.99(s,1H),8.38(d,J=7.8Hz,1H),8.05(t,J =5.8Hz,1H),7.92(d,J=2.3Hz,1H),7.78(d,J=9.2Hz,1H),7.73(s,1H),7.44(d,J=7 .8Hz,2H),7.38(d,J=7.9Hz,2H),7.34(d,J=2.0Hz,1H),7.20(d,J=2.4Hz,1H),6.94 -6.87(m,1H),6.65-6.56(m,1H),6.34-6.24(m,1H),5.13-5.03(m,2H),4.96-4.88( m,1H),4.62(d,J=13.6Hz,1H),4.56-4.49(m,3H),4.42(t,J=8.0Hz,1H),4.28(s,1H ),3.66-3.56(m,5H),3.17-3.10(m,1H),3.07-3.03(m,1H),3.01(s,3H),2.46(s,3H ),2.30-2.22(m,1H),2.14-2.07(m,1H),2.04-1.97(m,1H),1.84-1.76(m,1H),1.53 -1.44(m,2H),1.40-1.32(m,5H),1.26-1.16(m,14H),0.94(s,9H).LC-MS(ESI):m / z 1123.4[M+H] + .

[0344] Step 1.8. Synthesis of compound XZ1701-P2. The title compound (14.2 mg) was obtained from compounds 12 and 18-P2 using a general procedure. 1H NMR (600MHz, DMSO-d6) δ11.76(s,1H),8.91(s,1H),8.35(t,J=7.9Hz,1H),8.25-8.21(m,1H),7.87(d,J=4.1Hz,1H),7.67(d,J=3 .1Hz,1H),7.37(dd,J=8.3,2.4Hz,3H),7.33-7.28(m,3H),7.14-7.07(m,1H),6.90-6.82(m,1H),6.60-6.53(m,1H),6.36-6.24(m ,1H),5.69(s,1H),5.11-4.80(m,3H),4.59-4.35(m,5H),4.22(s,1H),3.92-3.83(m,2H),3.58-3.49(m,5H),3.47-3.30(m,3H), 2.94(s,3H),2.40-2.36(m,3H),2.00-1.96(m,1H),1.74-1.68(m,1H),1.28(d,J=7.0Hz,3H),0.92-0.83(m,9H).LC-MS(ESI):m / z 1027.4[M+H] + .

[0345] Step 1.9. Synthesis of compound XZ1701-P3. The title compound (13.0 mg) was obtained from compounds 12 and 18-P3 using a general procedure. 1H NMR(600MHz,DMSO-d6)δ11.88-11.79(m,1H),8.98(s,1H),8.43(d,J=7.7Hz, 1H),8.16(d,J=6.2Hz,1H),7.94(d,J=2.0Hz,1H),7.73(s,1H),7.46-7.42(m ,2H),7.42-7.33(m,4H),7.20(d,J=2.6Hz,1H),6.97-6.91(m,1H),6.66-6.6 0(m,1H),6.38-6.31(m,1H),5.14(d,J=3.5Hz,1H),5.07(d,J=16.6Hz,1H),4 .96-4.89(m,1H),4.66-4.60(m,1H),4.58-4.48(m,3H),4.48-4.42(m,1H),4 .29(s,1H),4.02-3.90(m,2H),3.65-3.60(m,6H),3.59-3.53(m,3H),3.44-3 .39(m,2H),3.32-3.24(m,2H),3.01(s,3H),2.46(s,3H),2.07-2.02(m,1H), 1.82-1.75(m,1H),1.37(d,J=7.0Hz,3H),0.96-0.92(m,9H).LC-MS(ESI):m / z 1071.4 [M+H] + .

[0346] Step 1.10. Synthesis of compound XZ1701-P4. The title compound (13.0 mg) was obtained from compounds 12 and 18-P4 using a general procedure. 1H NMR (600MHz, DMSO-d6) δ11.83(d,J=2.7Hz,1H),8.98(s,1H),8.44(d,J=7.7Hz,1H),8.19-8.13(m,1H),7.93(d,J=2.1Hz,1H),7.73(d,J=3.3Hz,1H),7 .46-7.34(m,6H),7.19(d,J=2.7Hz,1H),6.97-6.90(m,1H),6.65-6.59(m,1 H),6.37-6.30(m,1H),5.14(d,J=3.5Hz,1H),5.08-5.03(m,1H),4.93-4.88 (m,1H),4.62(d,J=13.5Hz,1H),4.58-4.48(m,3H),4.47-4.42(m,1H),4.2 9(s,1H),3.97(d,J=1.2Hz,2H),3.64-3.50(m,13H),3.39(s,2H),3.33-3.2 8(m,1H),3.25(d,J=12.8Hz,1H),3.00(s,3H),2.45(s,3H),2.07-2.03(m,1 H),1.81-1.75(m,1H),1.36(d,J=7.0Hz,3H),0.94(s,9H).LC-MS(ESI):m / z 1115.4[M+H] + .

[0347] Step 1.11. Synthesis of compound XZ1706-C10. Using compounds 12 and 19-C10 as starting materials, the title compound (13.0 mg) was obtained using a general procedure. 1H NMR(600MHz,DMSO-d6)δ11.83(s,1H),8.98(s,1H),8.56(d,J=6.5Hz,1H),8.05( t,J=5.8Hz,1H),7.92(d,J=2.1Hz,1H),7.84(d,J=9.3Hz,1H),7.72(s,1H),7.44- 7.37(m,4H),7.33(d,J=2.0Hz,1H),7.20(d,J=2.7Hz,1H),6.94-6.88(m,1H),6.6 2-6.58(m,1H),6.30-6.26(m,1H),5.13(d,J=3.5Hz,1H),5.06(d,J=16.6Hz,1H), 4.62(d,J=13.6Hz,1H),4.58-4.48(m,3H),4.48-4.39(m,2H),4.35(s,1H),4.26 -4.19(m,1H),3.69-3.64(m,2H),3.60(s,3H),3.18-3.09(m,1H),3.07-2.98(m,4 H),2.44(s,3H),2.29-2.23(m,1H),2.15-2.01(m,2H),1.94-1.88(m,1H),1.56-1 .42(m,2H),1.39-1.30(m,2H),1.28-1.14(m,10H),0.93(s,9H).LC-MS(ESI):m / z 1081.4[M+H] + .

[0348] Scheme 3. Synthesis of the new degradation agent XZ1707 series

[0349]

[0350] General Step 1: Compound 13 (1 equivalent), compound 18 (1.2 equivalent), and N,N-diisopropylethylamine (2 equivalent) were dissolved in dichloromethane (2 mL) and methanol (1 mL). The mixture was reacted overnight at room temperature, followed by the addition of sodium cyanoborohydride (3.0 equivalent) and a further reaction time of 1 hour. After the reaction was completed by LC-MS monitoring, the mixture was concentrated under reduced pressure. The residue was purified by preparative liquid chromatography to obtain the title compound.

[0351] Step 1.1. Synthesis of compound XZ1707-C8

[0352] The title compound (15.0 mg) was obtained from compounds 13 and 18-C8 using a general procedure. 1H NMR(600MHz,DMSO-d6)δ11.91(d,J=2.9Hz,1H),8.99(s,1H),8.84(s,1H),8.68(s ,1H),8.36(d,J=7.8Hz,1H),7.99(s,1H),7.79(d,J=9.3Hz,1H),7.74(s,1H),7.5 5(s,1H),7.44(d,J=8.0Hz,2H),7.39(d,J=7.9Hz,2H),7.26(d,J=2.6Hz,1H),7.1 4-7.07(m,1H),6.70-6.64(m,1H),6.32-6.24(m,1H),5.10(d,J=16.6Hz,1H),4.9 7-4.89(m,1H),4.64(d,J=13.6Hz,1H),4.55-4.50(m,3H),4.42(t,J=8.1Hz,1H), 4.29(s,2H),4.01-3.94(m,1H),3.63-3.60(m,4H),3.07(s,3H),2.99-2.91(m,2H ),2.46(s,3H),2.31-2.22(m,1H),2.16-1.99(m,2H),1.85-1.77(m,1H),1.64-1. 43(m,5H),1.38(d,J=7.0Hz,3H),1.30-1.23(m,7H),0.94(s,9H).LC-MS(ESI):m / z 1053.4[M+H] + .

[0353] Step 1.2. Synthesis of compound XZ1707-C9

[0354] The title compound (14.3 mg) was obtained from compounds 13 and 18-C9 using a general procedure. 1H NMR(600MHz,DMSO-d6)δ11.91(d,J=2.8Hz,1H),8.99(s,1H),8.82(s,1H),8.67(s,1H ),8.36(d,J=7.8Hz,1H),7.99(d,J=2.0Hz,1H),7.78(d,J=9.3Hz,1H),7.74(s,1H),7. 55(s,1H),7.44(d,J=8.1Hz,2H),7.39(d,J=8.0Hz,2H),7.26(d,J=2.6Hz,1H),7.14-7 .07(m,1H),6.71-6.64(m,1H),6.32-6.23(m,1H),5.10(d,J=16.6Hz,1H),4.96-4.89( m,1H),4.64(d,J=13.6Hz,1H),4.56-4.49(m,3H),4.42(t,J=8.0Hz,1H),4.32-4.21(m ,2H),4.02-3.95(m,1H),3.63-3.60(m,4H),3.07(s,3H),2.98(s,2H),2.46(s,3H),2. 31-2.22(m,1H),2.14-2.08(m,1H),2.05-1.99(m,1H),1.84-1.77(m,1H),1.60(s,2H) ,1.55-1.41(m,3H),1.38(d,J=7.0Hz,3H),1.26(s,9H),0.94(s,9H).LC-MS(ESI):m / z 1067.4[M+H] + .

[0355] Step 1.3. Synthesis of compound XZ1707-C10

[0356] The title compound (16.8 mg) was obtained from compounds 13 and 18-C10 using a general procedure. 1H NMR(600MHz,DMSO-d6)δ11.90(s,1H),8.99(s,1H),8.84(s,1H),8.74-8.63(m,1H),8 .37(d,J=7.9Hz,1H),7.99(s,1H),7.78(d,J=9.3Hz,1H),7.74(s,1H),7.55(s,1H),7. 44(d,J=8.0Hz,2H),7.39(d,J=8.0Hz,2H),7.26(d,J=2.7Hz,1H),7.14-7.08(m,1H),6 .70-6.65(m,1H),6.30-6.24(m,1H),5.10(d,J=16.6Hz,1H),4.97-4.89(m,1H),4.64( d,J=13.6Hz,1H),4.55-4.50(m,3H),4.42(t,J=8.1Hz,1H),4.31-4.23(m,2H),4.03-3 .95(m,1H),3.62-3.60(m,4H),3.07(s,3H),2.98-2.87(m,2H),2.46(s,3H),2.30-2.2 3(m,1H),2.14-2.07(m,1H),2.05-1.99(m,1H),1.84-1.77(m,1H),1.63-1.56(m,2H), 1.54-1.44(m,2H),1.38(d,J=7.0Hz,3H),1.26(s,12H),0.94(s,9H).LC-MS(ESI):m / z 1081.5[M+H] + .

[0357] Step 1.4. Synthesis of compound XZ1707-C11

[0358] The title compound (16.7 mg) was obtained from compounds 13 and 18-C11 using a general procedure. 1H NMR(600MHz,DMSO-d6)δ11.90(s,1H),8.99(s,1H),8.91(s,1H),8.76(s,1H),8.37(d, J=7.9Hz,1H),7.99(d,J=2.0Hz,1H),7.78(d,J=9.2Hz,1H),7.74(s,1H),7.56(d,J=1. 9Hz,1H),7.44(d,J=8.2Hz,2H),7.38(d,J=8.1Hz,2H),7.27-7.23(m,1H),7.15-7.03( m,1H),6.71-6.61(m,1H),6.34-6.22(m,1H),5.10(d,J=16.6Hz,1H),4.95-4.87(m,1H) ,4.63(d,J=13.6Hz,1H),4.56-4.48(m,3H),4.45-4.38(m,1H),4.27(d,J=13.5Hz,2H) ,4.01-3.95(m,1H),3.61-3.59(m,4H),3.06(s,3H),2.46(s,3H),2.29-2.22(m,1H),2 .13-2.06(m,1H),2.02(t,J=10.7Hz,1H),1.83-1.74(m,1H),1.59(s,2H),1.54-1.41( m,3H),1.38(d,J=7.0Hz,3H),1.27-1.23(m,13H),0.96-0.90(m,11H).LC-MS(ESI):m / z 1095.5[M+H] + .

[0359] Step 1.5. Synthesis of compound XZ1707-C12

[0360] The title compound (15.0 mg) was obtained from compounds 13 and 18-C12 using a general procedure. 1H NMR(600MHz,DMSO-d6)δ11.90(s,1H),8.99(s,1H),8.82(s,1H),8.68(s,1H), 8.37(d,J=7.8Hz,1H),7.99(s,1H),7.78(d,J=9.2Hz,1H),7.74(s,1H),7.55(s ,1H),7.44(d,J=8.0Hz,2H),7.39(d,J=8.0Hz,2H),7.25(d,J=2.7Hz,1H),7.12 -7.06(m,1H),6.69-6.64(m,1H),6.31-6.25(m,1H),5.10(d,J=16.5Hz,1H),4. 95-4.89(m,1H),4.63(d,J=13.7Hz,1H),4.54-4.49(m,3H),4.42(t,J=8.1Hz, 1H),4.29(s,2H),4.02-3.94(m,1H),3.61(s,4H),3.07(s,3H),2.95(s,2H),2. 46(s,3H),2.30-2.23(m,1H),2.13-1.98(m,2H),1.83-1.77(m,1H),1.59(s,2H ),1.53-1.37(m,6H),1.26-1.23(m,13H),0.96-0.92(m,11H).LC-MS(ESI):m / z 1109.5 [M+H] + .

[0361] Scheme 4. Synthesis of the new degradation agent XZ1708 series

[0362]

[0363] General Step 1: Compound 15 (1 equivalent), Compound 18 (1.2 equivalent), and N,N-diisopropylethylamine (4 equivalent) were dissolved in dimethyl sulfoxide (2 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.5 equivalent) was added and reacted at room temperature for 30 min. After the reaction was completed as monitored by LC-MS, the title compound was purified by preparative liquid chromatography.

[0364] Step 1.1. Synthesis of compound XZ1708-C6. Using compounds 15 and 18-C6 as starting materials, the title compound (12.0 mg) was obtained using a general procedure. 1H NMR (600MHz, DMSO-d6) δ11.81(s,1H),8.99(d,J=2.2Hz,1H),8.37(d,J=7.7H z,1H),7.80-7.73(m,3H),7.66-7.64(m,1H),7.46-7.42(m,2H),7.38(d,J=7 .9Hz,2H),7.29(s,1H),7.22(s,1H),7.05-6.95(m,1H),6.66-6.60(m,1H),6 .29-6.18(m,1H),5.19-5.09(m,2H),4.94-4.89(m,1H),4.59-4.50(m,2H),4. 45-4.37(m,3H),4.29(s,1H),3.64-3.57(m,5H),3.02-2.94(m,5H),2.86-2. 77(m,1H),2.69-2.60(m,1H),2.47-2.43(m,3H),2.41-2.33(m,1H),2.29-2.1 9(m,2H),2.16-2.10(m,1H),2.06-1.99(m,1H),1.83-1.76(m,1H),1.52-1.4 2(m,2H),1.41-1.29(m,5H),1.26-1.14(m,2H),0.93(s,9H).LC-MS(ESI):m / z 1067.4 [M+H] + .

[0365] Step 1.2. Synthesis of compound XZ1708-C7. Using compounds 15 and 18-C7 as starting materials, the title compound (12.0 mg) was obtained using a general procedure. 1H NMR (600MHz, DMSO-d6) δ11.82(d,J=2.7Hz,1H),8.98(s,1H),8.37(d,J=7.8Hz, 1H),7.78(d,J=9.3Hz,1H),7.75-7.72(m,2H),7.64(s,1H),7.45-7.41(m,2H), 7.39-7.36(m,2H),7.28(d,J=2.0Hz,1H),7.21(d,J=2.6Hz,1H),7.02-6.98(m, 1H),6.64-6.58(m,1H),6.27-6.20(m,1H),5.16(d,J=16.5Hz,1H),5.12-5.08(m ,1H),4.96-4.88(m,1H),4.58-4.50(m,2H),4.44-4.36(m,3H),4.30-4.26(m,1 H),3.63-3.56(m,5H),3.05-2.93(m,5H),2.84-2.77(m,1H),2.67-2.59(m,1H), 2.45(s,3H),2.40-2.34(m,1H),2.27-2.20(m,2H),2.13-2.07(m,1H),2.03-1. 97(m,1H),1.83-1.76(m,1H),1.48-1.20(m,11H),0.93(s,9H).LC-MS(ESI):m / z 1081.4[M+H] + .

[0366] Step 1.3. Synthesis of compound XZ1708-C8. Using compounds 15 and 18-C8 as starting materials, the title compound (12.0 mg) was obtained using a general procedure. 1H NMR(600MHz,DMSO-d6)δ11.82(d,J=2.8Hz,1H),8.98(s,1H),8.37(d,J=7.8 Hz,1H),7.79-7.71(m,3H),7.64(s,1H),7.43(d,J=7.9Hz,2H),7.38(d,J=8. 0Hz,2H),7.29(d,J=2.1Hz,1H),7.21(d,J=2.7Hz,1H),7.06-6.94(m,1H),6 .67-6.61(m,1H),6.25-6.19(m,1H),5.19-5.09(m,2H),4.96-4.88(m,1H),4 .57-4.50(m,2H),4.44-4.36(m,3H),4.31-4.26(m,1H),3.64-3.57(m,5H), 2.97(s,5H),2.89-2.77(m,1H),2.70-2.59(m,1H),2.45(s,2H),2.40-2.33( m,1H),2.27-2.19(m,2H),2.13-2.07(m,2H),2.04-1.98(m,1H),1.83-1.74 (m,1H),1.53-1.30(m,7H),1.27-1.16(m,6H),0.93(s,9H).LC-MS(ESI):m / z 1095.4 [M+H] + .

[0367] Step 1.4. Synthesis of compound XZ1708-C9. Using compounds 15 and 18-C9 as starting materials, the title compound (15.4 mg) was obtained using a general procedure. 1H NMR (600MHz, DMSO-d6) δ11.81(s,1H),8.99(d,J=3.0Hz,1H),8.37(d,J=7.7Hz,1H),7.81-7.72(m,3H),7.65(d,J=3.1Hz,1H),7.46-7.36(m,4H ),7.25(d,J=44.8Hz,2H),7.06-6.96(m,1H),6.67-6.59(m,1H),6.28- 6.19(m,1H),5.22-5.08(m,2H),4.96-4.90(m,1H),4.60-4.50(m,2H),4 .47-4.36(m,3H),4.28(s,1H),3.66-3.56(m,5H),3.03-2.93(m,5H),2 .84-2.77(m,1H),2.67-2.61(m,1H),2.48-2.44(m,3H),2.41-2.34(m,1 LC-MS(ESI):m / z 1109.4[M+H] + .

[0368] Step 1.5. Synthesis of compound XZ1708-C10. Using compounds 15 and 18-C10 as starting materials, the title compound (8.7 mg) was obtained using a general procedure. 1H NMR (600MHz, DMSO-d6) δ11.82(s,1H),8.99(d,J=3.2Hz,1H),8.37(d,J=7.7Hz,1 H),7.75(t,J=16.1Hz,3H),7.65(d,J=3.1Hz,1H),7.44(d,J=7.6Hz,2H),7.38(d ,J=6.8Hz,2H),7.36-7.28(m,1H),7.22(s,1H),7.05-6.94(m,1H),6.62(t,J=8. 7Hz,1H),6.27-6.18(m,1H),5.17(d,J=16.6Hz,1H),5.10(d,J=3.8Hz,1H),4.96 -4.89(m,1H),4.60-4.49(m,2H),4.47-4.36(m,3H),4.28(s,1H),3.64-3.57(m, 5H),3.06-2.94(m,5H),2.85-2.78(m,1H),2.66-2.59(m,1H),2.48-2.45(m,3H) ,2.43-2.33(m,1H),2.24(s,2H),2.13-2.08(m,1H),2.05-1.98(m,1H),1.83-1. 76(m,1H),1.51-1.31(m,7H),1.27-1.14(m,10H),0.94(s,9H).LC-MS(ESI):m / z 1123.4[M+H] + .

[0369] Scheme 5. Synthesis of new degradation agents XZ1729 series and XZ1733 series

[0370]

[0371] General Step 1: Compound 17 or 13 (1 equivalent), compound 19 (1.2 equivalent), and N,N-diisopropylethylamine (2 equivalent) were dissolved in dichloromethane (2 mL) and methanol (1 mL). The mixture was reacted overnight at room temperature, followed by the addition of sodium cyanoborohydride (3 equivalent) and a further reaction time of 1 hour. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue was purified by preparative liquid chromatography to obtain the title compound.

[0372] Step 1.1. Synthesis of compound XZ1729-C11. Using compounds 17 and 19-C11 as starting materials, the title compound (19.0 mg) was obtained via a general procedure. LC-MS (ESI): m / z 1082.4 [M+H] + .

[0373] Step 1.2. Synthesis of compound XZ1733-C10. Using compounds 13 and 19-C10 as starting materials, the title compound (10.2 mg) was obtained using a general procedure. 1 H NMR(600MHz,DMSO-d6)δ11.90(s,1H),8.99(s,1H),8.85(s,1H),8.71(s,1H),8 .56(t,J=6.1Hz,1H),7.99(s,1H),7.84(d,J=9.4Hz,1H),7.74(s,1H),7.55(s,1 H),7.43(d,J=8.1Hz,2H),7.39(d,J=8.0Hz,2H),7.25(d,J=2.8Hz,1H),7.13-7 .06(m,1H),6.70-6.64(m,1H),6.31-6.24(m,1H),5.10(d,J=16.6Hz,1H),4.63( d,J=13.6Hz,1H),4.57-4.49(m,3H),4.47-4.40(m,2H),4.36(s,1H),4.31-4.1 9(m,2H),4.01-3.95(m,1H),3.69-3.60(m,5H),3.06(s,3H),2.97-2.86(m,2H), 2.45(s,3H),2.31-2.23(m,1H),2.15-2.07(m,1H),2.07-2.01(m,1H),1.95-1. 88(m,1H),1.62-1.44(m,4H),1.32-1.20(m,11H),0.94(s,9H).LC-MS(ESI):m / z 1067.4 [M+H] + .

[0374] Step 1.3. Synthesis of compound XZ1733-C11. The title compound (11.8 mg) was obtained from compounds 13 and 19-C11 using a general procedure. 1H NMR(600MHz,DMSO-d6)δ11.90(d,J=2.7Hz,1H),8.99(s,1H),8.82(s,1H),8.68(s,1H ),8.56(t,J=6.2Hz,1H),7.99(d,J=1.9Hz,1H),7.83(d,J=9.3Hz,1H),7.74(s,1H),7 .55(d,J=2.0Hz,1H),7.43(d,J=8.2Hz,2H),7.39(d,J=8.3Hz,2H),7.25(d,J=2.7Hz, 1H),7.13-7.06(m,1H),6.70-6.64(m,1H),6.31-6.24(m,1H),5.10(d,J=16.6Hz,1H), 4.63(d,J=13.6Hz,1H),4.57-4.49(m,3H),4.47-4.40(m,2H),4.36(s,1H),4.25(ddd ,J=21.5,12.7,5.2Hz,2H),4.01-3.95(m,1H),3.70-3.60(m,6H),3.07(s,3H),2.97-2 .87(m,2H),2.45(s,3H),2.31-2.23(m,1H),2.15-2.07(m,1H),2.07-2.00(m,1H),1. 95-1.88(m,1H),1.62-1.41(m,4H),1.30-1.22(m,12H),0.94(s,9H).LC-MS(ESI):m / z 1081.5[M+H] + .

[0375] Scheme 6. Synthesis of new degradation agents XZ1713 and XZ1714 series

[0376]

[0377] General Step 1: Compound 20 or 23 (1 equivalent), the corresponding acid 21 (1.2 equivalent), and N,N-diisopropylethylamine (4 equivalent) were dissolved in dichloromethane (20 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.5 equivalent) was added, and the mixture was reacted at room temperature for 30 minutes. After the reaction was complete as monitored by LC-MS, the mixture was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain an intermediate. This intermediate was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (2 mL) was added. The mixture was reacted at room temperature for 1 hour. After the reaction was complete as monitored by LC-MS, the mixture was concentrated under reduced pressure, and the residue was slurried in diethyl ether to give the corresponding amino compound 22 or 24.

[0378] General Step 2: Compound 12 (1 equivalent), the corresponding amino compound 22 or 24 (1.2 equivalent), and N,N-diisopropylethylamine (4 equivalent) were dissolved in dimethyl sulfoxide (2 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.5 equivalent) were added. The mixture was reacted at room temperature for 30 minutes. After the reaction was completed as monitored by LC-MS, the title compound was purified by preparative liquid chromatography.

[0379] Step 1.1. Synthesis of Compound 22-C2. Using compounds 20 and 21-C2 as starting materials, the title compound (153 mg) was obtained via the general procedure 1. LC-MS (ESI): m / z 400.1 [M+H] + .

[0380] Step 1.2. Synthesis of Compound 22-C4. Using compounds 20 and 21-C4 as starting materials, the title compound (176 mg) was obtained via the general procedure 1. LC-MS (ESI): m / z 428.2 [M+H] + .

[0381] Step 1.3. Synthesis of Compound 22-C6. Using compounds 20 and 21-C6 as starting materials, the title compound (147 mg) was obtained via general step 1. LC-MS (ESI): m / z 456.2 [M+H] + .

[0382] Step 1.4. Synthesis of Compound 22-C8. Using compounds 20 and 21-C8 as starting materials, the title compound (191 mg) was obtained via general step 1. LC-MS (ESI): m / z 484.3 [M+H] + .

[0383] Step 1.5. Synthesis of Compound 22-C10. Using compounds 20 and 21-C10 as starting materials, the title compound (97 mg) was obtained via general step 1. LC-MS (ESI): m / z 512.3 [M+H] + .

[0384] Step 1.6. Synthesis of Compound 22-P2. Using compounds 20 and 21-P2 as starting materials, the title compound (112 mg) was obtained via general step 1. LC-MS (ESI): m / z 458.2 [M+H] + .

[0385] Step 1.7. Synthesis of Compound 22-P4. Using compounds 20 and 21-P4 as starting materials, the title compound (165 mg) was obtained via general step 1. LC-MS (ESI): m / z 546.3 [M+H] + .

[0386] Step 1.8. Synthesis of Compound 24-C2. Using compounds 23 and 21-C2 as starting materials, the title compound (171 mg) was obtained via general step 1. LC-MS (ESI): m / z 331.1 [M+H] + .

[0387] Step 1.9. Synthesis of Compound 24-C3. Using compounds 23 and 21-C3 as starting materials, the title compound (159 mg) was obtained via general step 1. LC-MS (ESI): m / z 345.2 [M+H] + .

[0388] Step 1.10. Synthesis of Compound 24-C4. Using compounds 23 and 21-C4 as starting materials, the title compound (163 mg) was obtained via general step 1. LC-MS (ESI): m / z 359.2 [M+H] + .

[0389] Step 1.11. Synthesis of Compound 24-C6. Using compounds 23 and 21-C6 as starting materials, the title compound (202 mg) was obtained via general step 1. LC-MS (ESI): m / z 387.2 [M+H] + .

[0390] Step 1.12. Synthesis of Compound 24-P2. Using compounds 23 and 21-P2 as starting materials, the title compound (101 mg) was obtained via general step 1. LC-MS (ESI): m / z 389.2 [M+H] + .

[0391] Step 1.13. Synthesis of Compound 24-P4. Using compounds 23 and 21-P4 as starting materials, the title compound (93 mg) was obtained via general step 1. LC-MS (ESI): m / z 477.3 [M+H] + .

[0392] Step 1.14. Synthesis of Compound 24-A. Using compounds 23 and 21-A as starting materials, the title compound (110 mg) was obtained via general step 1. LC-MS (ESI): m / z 345.2 [M+H] + .

[0393] Step 1.15. Synthesis of Compound 24-B. Using compounds 23 and 21-B as starting materials, the title compound (80 mg) was obtained via general step 1. LC-MS (ESI): m / z 345.2 [M+H] + .

[0394] Step 1.16. Synthesis of Compound 24-C. Using compounds 23 and 21-C as starting materials, the title compound (102 mg) was obtained via general step 1. LC-MS (ESI): m / z 371.2 [M+H] + .

[0395] Step 1.17. Synthesis of Compound 24-D. Using compounds 23 and 21-D as starting materials, the title compound (76 mg) was obtained via general step 1. LC-MS (ESI): m / z 371.2 [M+H] + .

[0396] Step 2.1. Synthesis of compound XZ1713-C2. Using compounds 12 and 22-C2 as starting materials, the title compound (13.0 mg) was obtained by general step 2. 1 H NMR (600MHz, DMSO-d6) δ11.86(d,J=2.7Hz,1H),11.09(s,1H),8.50(t,J=5.0Hz,1H),7.96(d,J=2.1Hz,1H),7.74-7.69(m,2H) ,7.59(d,J=2.0Hz,1H),7.36(d,J=2.3Hz,1H),7.29-7.24(m,2H),6.93-6.86(m,1H),6.65-6.59(m,1H),6.42-6.35(m,1H),5. 16(d,J=16.6Hz,1H),5.08(dd,J=12.8,5.5Hz,1H),4.66(d,J=13.5Hz,1H),4.62-4.51(m,2H),4.17-4.06(m,2H),3.66-3.60( m,6H),3.56-3.50(m,4H),3.17(s,1H),3.02(s,3H),2.92-2.84(m,1H),2.63-2.53(m,2H),2.06-1.99(m,1H).LC-MS(ESI):m / z 881.2[M+H] + .

[0397] Step 2.2. Synthesis of compound XZ1713-C4. Using compounds 12 and 22-C4 as starting materials, the title compound (11.6 mg) was obtained by general step 2. 1H NMR (600MHz, DMSO-d6) δ11.82(d,J=2.6Hz,1H),11.09(s,1H),8.12(t,J=5.8Hz,1H),7.92(d,J=2.1Hz,1H),7.73(s,1H),7.71 (d,J=8.5Hz,1H),7.34(dd,J=6.1,2.1Hz,2H),7.24(dd,J=8.5,2.3Hz,1H),7.20(d,J=2.7Hz,1H),6.96-6.89(m,1H),6.66-6.5 9(m,1H),6.34-6.27(m,1H),5.10-5.04(m,2H),4.65-4.49(m,3H),3.65-3.59(m,6H),3.23-3.09(m,6H),3.01(s,3H),2.90-2. 85(m,1H),2.64-2.56(m,2H),2.31(q,J=7.0Hz,2H),2.05-1.98(m,1H),1.70-1.59(m,2H),1.29-1.22(m,1H).LC-MS(ESI):m / z 909.3[M+H] + .

[0398] Step 2.3. Synthesis of compound XZ1713-C6. Using compounds 12 and 22-C6 as starting materials, the title compound (6.0 mg) was obtained via general step 2. 1H NMR (600MHz, DMSO-d6) δ11.83(d,J=2.7Hz,1H),11.09(s,1H),8.07(t,J=5.8Hz,1H),7.92(d,J=2.0Hz,1H),7.74-7.68(m,2H),7.37-7.3 2(m,2H),7.25(dd,J=8.7,2.4Hz,1H),7.21(d,J=2.7Hz,1H),6.97-6.89(m,1H),6.65-6.59(m,1H),6.34-6.26(m,1H),5.11-5.05(m,2H) ,4.62(d,J=13.5Hz,1H),4.57-4.48(m,2H),3.61(s,6H),3.52-3.46(m,4H),3.20-3.13(m,2H),3.08-3.01(m,4H),2.92-2.85(m,1H),2. 64-2.55(m,2H),2.32(t,J=7.5Hz,2H),2.03(d,J=6.4Hz,1H),1.54-1.47(m,2H),1.43-1.36(m,2H),1.27-1.23(m,2H).LC-MS(ESI):m / z 937.3[M+H] + .

[0399] Step 2.4. Synthesis of compound XZ1713-C8. Using compounds 12 and 22-C8 as starting materials, the title compound (8.5 mg) was obtained via general step 2. 1H NMR (600MHz, DMSO-d6) δ11.83(d,J=2.7Hz,1H),11.08(s,1H),8.06(t,J=5.8Hz,1H),7.92(d,J=2.0Hz,1H),7.74-7.68(m,2H),7.34( d,J=2.2Hz,2H),7.26-7.19(m,2H),6.96-6.89(m,1H),6.64-6.58(m,1H),6.33-6.26(m,1H),5.11-5.04(m,2H),4.62(d,J=13.6Hz,1 H),4.57-4.49(m,2H),3.64-3.59(m,7H),3.53-3.49(m,3H),3.20-3.11(m,2H),3.07-3.03(m,1H),3.01(s,3H),2.92-2.84(m,1H),2 .62-2.53(m,2H),2.36(t,J=7.5Hz,2H),2.06-2.00(m,1H),1.55-1.48(m,2H),1.40-1.33(m,2H),1.29-1.20(m,6H).LC-MS(ESI):m / z 965.3[M+H] + .

[0400] Step 2.5. Synthesis of compound XZ1713-P2. Using compounds 12 and 22-P2 as starting materials, the title compound (9.2 mg) was obtained via general step 2. 1 H NMR (600MHz, DMSO-d6) δ11.81(s,1H),11.08(s,1H),8.09(t,J=5.3Hz,1H),7.93(s,1H),7.72(s,1H),7.69 (d,J=8.5Hz,1H),7.37(s,1H),7.31(s,1H),7.22-7.18(m,2H),6.97-6.90(m,1H),6.65-6.60(m,1H),6.37 -6.30(m,1H),5.11-5.03(m,2H),4.63(d,J=13.6Hz,1H),4.51(d,J=14.7Hz,2H),3.62(d,J=16.7Hz,15H), 3.27-3.20(m,2H),3.01(s,3H),2.92-2.85(m,1H),2.65-2.54(m,4H),2.06-2.01(m,1H).LC-MS(ESI):m / z 939.3[M+H] + .

[0401] Step 2.6. Synthesis of compound XZ1713-P4. Using compounds 12 and 22-P4 as starting materials, the title compound (6.5 mg) was obtained via general step 2. 1 H NMR (600MHz, DMSO-d6) δ11.83(d,J=2.7Hz,1H),11.08(s,1H),8.14(t,J=5.6Hz,1H),7.93(d,J=2.1Hz,1H),7.73(s, 1H),7.69(d,J=8.5Hz,1H),7.38-7.32(m,2H),7.24-7.18(m,2H),6.97-6.89(m,1H),6.66-6.59(m,1H),6.38-6.31( m,1H),5.11-5.03(m,2H),4.62(d,J=13.6Hz,1H),4.52(dd,J=19.5,15.0Hz,2H),3.67-3.58(m,10H),3.53-3.47(m, 11H),3.33-3.20(m,4H),3.01(s,3H),2.88(d,J=12.6Hz,1H),2.63-2.54(m,4H),2.05-1.99(m,1H).LC-MS(ESI):m / z 1027.3[M+H] + .

[0402] Step 2.7. Synthesis of compound XZ1714-C2. Using compounds 12 and 24-C2 as starting materials, the title compound (11.0 mg) was obtained by general step 2. 1 H NMR (600MHz, DMSO-d6) δ11.86(d,J=2.7Hz,1H),10.78(s,1H),8.47(t,J=5.1Hz,1H),7.95(d,J=2.1Hz,1H),7.72(s, 1H),7.59(d,J=2.0Hz,1H),7.26(d,J=2.7Hz,1H),7.09(d,J=8.6Hz,2H),6.96-6.86(m,3H),6.65-6.59(m,1H),6.42 -6.35(m,1H),5.15(d,J=16.6Hz,1H),4.69-4.51(m,3H),4.16-4.05(m,2H),3.75(dd,J=11.2,4.9Hz,1H),3.65-3.5 6(m,7H),3.19-3.10(m,5H),3.01(s,3H),2.69-2.62(m,1H),2.19-2.10(m,1H),2.05-1.98(m,1H).LC-MS(ESI):m / z 812.3[M+H]+ .

[0403] Step 2.8. Synthesis of compound XZ1714-C4. Using compounds 12 and 24-C4 as starting materials, the title compound (12.5 mg) was obtained by general step 2. 1 H NMR (600MHz, DMSO-d6) δ11.83(d,J=2.8Hz,1H),10.77(s,1H),8.11(t,J=5.7Hz,1H),7.92(d,J=2.1Hz,1H),7.73(s,1H),7.33(d,J=2. 0Hz,1H),7.20(d,J=2.6Hz,1H),7.10-7.05(m,2H),6.95-6.88(m,3H),6.65-6.59(m,1H),6.34-6.26(m,1H),5.06(d,J=16.5Hz,1H),4 .64-4.49(m,3H),3.74(dd,J=11.2,4.9Hz,1H),3.62-3.59(m,4H),3.52-3.47(m,3H),3.23-3.18(m,2H),3.11-3.07(m,4H),3.01(s,3 LC-MS(ESI):m / z 840.3[M+H] + .

[0404] Step 2.9. Synthesis of compound XZ1714-C6. Using compounds 12 and 24-C6 as starting materials, the title compound (9.8 mg) was obtained via general step 2. 1H NMR (600MHz, DMSO-d6) δ11.83(s,1H),10.78(s,1H),8.07(t,J=5.9Hz,1H),7.92(s,1H),7.73(s,1H),7.34(s,1H),7.21(s,1H),7.08(d,J=8.0Hz ,2H),6.95-6.89(m,3H),6.65-6.59(m,1H),6.34-6.27(m,1H),5.08(d, J=16.5Hz,1H),4.62(d,J=13.6Hz,1H),4.58-4.49(m,2H),3.74(dd,J=11 .0,4.9Hz,1H),3.63-3.57(m,7H),3.19-3.12(m,3H),3.10-3.06(m,2H) ,3.01(s,3H),2.67-2.61(m,1H),2.56-2.53(m,1H),2.47-2.44(m,1H),2 .31(t,J=7.6Hz,2H),2.16-2.10(m,1H),2.04-1.98(m,1H),1.54-1.46(m ,2H),1.42-1.36(m,2H),1.26-1.23(m,2H).LC-MS(ESI):m / z868.3[M+H] + .

[0405] Step 2.10. Synthesis of compound XZ1714-P2. Using compounds 12 and 24-P2 as starting materials, the title compound (6.2 mg) was obtained by general step 2. 1H NMR (600MHz, DMSO-d6) δ11.83(d,J=2.6Hz,1H),10.78(s,1H),8.12(t,J=5.7Hz,1H),7.93(d,J=2.0Hz,1H),7.73(s,1H),7.37(d,J= 1.9Hz,1H),7.20(d,J=2.7Hz,1H),7.07(d,J=8.3Hz,2H),6.97-6.89(m,3H),6.66-6.60(m,1H),6.38-6.31(m,1H),5.07(d,J=16.6H z,1H),4.62(d,J=13.6Hz,1H),4.57-4.48(m,2H),3.77-3.71(m,1H),3.67-3.54(m,10H),3.26-3.19(m,2H),3.14-3.10(m,2H),3.0 7-3.04(m,2H),3.01(s,3H),2.67-2.60(m,3H),2.49-2.44(m,2H),2.12(dd,J=18.2,7.5Hz,1H),2.04-1.98(m,1H).LC-MS(ESI):m / z 870.3[M+H] + .

[0406] Step 2.11. Synthesis of compound XZ1714-P4. Using compounds 12 and 24-P4 as starting materials, the title compound (9.9 mg) was obtained by general step 2. 1H NMR (600MHz, DMSO-d6) δ11.84(d,J=2.6Hz,1H),10.78(s,1H),8.14(t,J=5.6Hz,1H),7.93(d,J=2.0Hz,1H),7.73(s,1H),7.37(d,J=1.9Hz,1H ),7.19(d,J=2.6Hz,1H),7.07(d,J=8.3Hz,2H),6.97-6.89(m,3H),6.66-6.59(m,1H),6.39-6.31(m,1H),5.06(d,J=16.5Hz,1H),4.62(d,J=1 3.6Hz,1H),4.57-4.48(m,2H),3.74(dd,J=11.1,4.9Hz,1H),3.65-3.56(m,9H),3.52-3.49(m,7H),3.34-3.21(m,5H),3.10(t,J=4.9Hz,2H), 3.06(t,J=5.3Hz,2H),3.00(s,3H),2.67-2.58(m,3H),2.48-2.45(m,1H),2.16-2.09(m,1H),2.04-1.98(m,1H).LC-MS(ESI):m / z958.3[M+H] + .

[0407] Scheme 7. Synthesis of the new degradation agents XZ1737, XZ1738 and XZ1739 series

[0408]

[0409] General Step 1: Compounds 25 / 27 / 29 (1 equivalent), the corresponding acid 21 (1.2 equivalent), and N,N-diisopropylethylamine (4 equivalent) were dissolved in dichloromethane (20 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.5 equivalent) was added, and the mixture was reacted at room temperature for 30 minutes. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain an intermediate. This intermediate was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (2 mL) was added. The mixture was reacted at room temperature for 1 hour. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure, and the residue was slurried in diethyl ether to give the corresponding amino compounds 26 / 28 / 30.

[0410] General Step 2: Compound 13 (1 equivalent), the corresponding amino compounds 26 / 28 / 30 (1.2 equivalents), and N,N-diisopropylethylamine (2 equivalents) were dissolved in dichloromethane (2 mL) and methanol (1 mL). The mixture was reacted overnight at room temperature, followed by the addition of sodium cyanoborohydride (3 equivalents) and a further reaction time of 1 hour. After the reaction was completed by LC-MS monitoring, the mixture was concentrated under reduced pressure, and the residue was purified by preparative liquid chromatography to obtain the title compound.

[0411] Step 1.1. Synthesis of Compound 26-C2. Using compounds 25 and 21-C2 as starting materials, the title compound (110 mg) was obtained via general step 1. LC-MS (ESI): m / z 390.2 [M+H] + .

[0412] Step 1.2. Synthesis of Compound 26-C3. Using compounds 25 and 21-C3 as starting materials, the title compound (123 mg) was obtained via the general procedure 1. LC-MS (ESI): m / z 404.2 [M+H] + .

[0413] Step 1.3. Synthesis of Compound 26-C4. Using compounds 25 and 21-C4 as starting materials, the title compound (212 mg) was obtained via general step 1. LC-MS (ESI): m / z 418.2 [M+H] + .

[0414] Step 1.4. Synthesis of Compound 28-C2. Using compounds 27 and 21-C2 as starting materials, the title compound (91 mg) was obtained via general step 1. LC-MS (ESI): m / z 385.2 [M+H] + .

[0415] Step 1.5. Synthesis of Compound 28-C3. Using compounds 27 and 21-C3 as starting materials, the title compound (103 mg) was obtained via general step 1. LC-MS (ESI): m / z 399.2 [M+H] + .

[0416] Step 1.6. Synthesis of Compound 28-C4. Using compounds 27 and 21-C4 as starting materials, the title compound (161 mg) was obtained via general step 1. LC-MS (ESI): m / z 413.2 [M+H] + .

[0417] Step 1.7. Synthesis of Compound 30-C2. Using compounds 29 and 21-C2 as starting materials, the title compound (81 mg) was obtained via general step 1. LC-MS (ESI): m / z 332.2 [M+H]+ .

[0418] Step 2.1. Synthesis of compound XZ1737-C2. Using compounds 13 and 26-C2 as starting materials, the title compound (10.4 mg) was obtained by general step 2. 1 H NMR (600MHz, DMSO-d6) δ11.90(d,J=2.8Hz,1H),10.36(s,1H),9.24(s,2H),7.98(s,1H),7.72(s,1H),7.61(s,1H),7.46(dd,J =8.5,2.2Hz,1H),7.41(d,J=2.2Hz,1H),7.28(d,J=2.7Hz,1H),7.20(d,J=8.6Hz,1H),7.10-7.03(m,1H),6.70-6.64(m,1H),6 .31-6.24(m,1H),5.13(d,J=16.5Hz,1H),4.65(d,J=13.6Hz,1H),4.54(dd,J=25.9,15.1Hz,2H),4.39-4.33(m,1H),4.13(s,2 H),3.98(d,J=13.6Hz,1H),3.87(s,3H),3.65-3.46(m,13H),3.07(s,3H),2.70(t,J=6.7Hz,2H).LC-MS(ESI):m / z857.3[M+H] + .

[0419] Step 2.2. Synthesis of compound XZ1737-C3. Using compounds 13 and 26-C3 as starting materials, the title compound (9.0 mg) was obtained via general step 2. 1H NMR (600MHz, DMSO-d6) δ11.91(d,J=2.7Hz,1H),10.36(s,1H),8.73(d,J=76.7Hz,2H),8.01(d,J=2.0Hz,1H),7.74(s,1H),7.56(d,J=2.0Hz,1H),7. 45(dd,J=8.5,2.1Hz,1H),7.40(d,J=2.1Hz,1H),7.27(d,J=2.6Hz,1H),7. 19(d,J=8.6Hz,1H),7.14-7.07(m,1H),6.71-6.64(m,1H),6.29(td,J=9.7 ,5.8Hz,1H),5.13(d,J=16.5Hz,1H),4.65(d,J=13.6Hz,1H),4.52(t,J=13 .8Hz,2H),4.26-4.21(m,1H),4.06-4.00(m,1H),3.87(s,3H),3.64-3.59( m,6H),3.58-3.54(m,3H),3.52-3.49(m,3H),3.25-3.17(m,3H),3.06(s,3 H),2.83-2.78(m,2H),2.70(t,J=6.7Hz,2H).LC-MS(ESI):m / z871.3[M+H] + .

[0420] Step 2.3. Synthesis of compound XZ1737-C4. Using compounds 13 and 26-C4 as starting materials, the title compound (10.1 mg) was obtained by general step 2. 1H NMR (600MHz, DMSO-d6) δ11.91(d,J=2.8Hz,1H),10.35(s,1H),8.86(d,J=66.4Hz,2H),8.00(d,J=2.0Hz,1H),7.74(s,1H),7.55(d,J=2.0Hz,1H ),7.44(dd,J=8.5,2.2Hz,1H),7.39(d,J=2.2Hz,1H),7.26(d,J=2.6Hz,1H),7.19(d,J=8.5Hz,1H),7.13-7.06(m,1H),6.70-6.64(m,1H),6.32 -6.25(m,1H),5.11(d,J=16.6Hz,1H),4.64(d,J=13.6Hz,1H),4.54-4. 48(m,2H),4.30-4.23(m,1H),4.03-3.97(m,1H),3.86(s,3H),3.64-3.5 8(m,6H),3.55-3.46(m,7H),3.07(s,3H),3.04-2.93(m,2H),2.69(t,J=6.7Hz,2H),2.47(d,J=6.7Hz,2H),1.91-1.83(m,2H).LC-MS(ESI):m / z 885.3[M+H] + .

[0421] Step 2.4. Synthesis of compound XZ1738-C2. Using compounds 13 and 28-C2 as starting materials, the title compound (8.4 mg) was obtained via general step 2. 1H NMR(600MHz,DMSO-d6)δ11.90(t,J=2.2Hz,1H),10.55(s,1H),9.23(s,2H),7.99(d ,J=2.0Hz,1H),7.73(s,1H),7.64(d,J=1.9Hz,1H),7.59(dd,J=8.5,6.0Hz,1H),7. 40(d,J=4.0Hz,1H),7.30-7.26(m,1H),7.10-7.04(m,1H),7.03-7.01(m,1H),6.71 -6.64(m,1H),6.32-6.25(m,1H),5.15(dd,J=16.6,8.3Hz,1H),4.66(d,J=13.6Hz, 1H),4.61-4.51(m,3H),4.43-4.36(m,1H),4.19-4.09(m,2H),4.03-3.99(m,1H),3 .97(d,J=2.8Hz,3H),3.91(t,J=6.7Hz,2H),3.74(d,J=13.4Hz,1H),3.61(s,3H),3 .22-3.17(m,1H),3.07(s,3H),3.02-2.97(m,1H),2.85-2.79(m,1H),2.76(t,J=6. 7Hz,2H),1.96-1.87(m,2H),1.73-1.65(m,1H),1.61-1.54(m,1H).LC-MS(ESI):m / z 852.3[M+H] + .

[0422] Step 2.5. Synthesis of compound XZ1738-C3. Using compounds 13 and 28-C3 as starting materials, the title compound (8.4 mg) was obtained via general step 2. 1H NMR (600MHz, DMSO-d6) δ11.91(d,J=2.7Hz,1H),10.55(s,1H),8.75(d,J=60.3Hz,2H),8.02(s,1H),7.75(s,1H),7.61-7.55(m,2H),7.42(d,J=3.8Hz ,1H),7.27(s,1H),7.14-7.07(m,1H),7.07-7.00(m,1H),6.71-6.65(m,1H ),6.34-6.27(m,1H),5.15(dd,J=16.6,7.3Hz,1H),4.66(d,J=13.6Hz,1H) ,4.60(d,J=12.4Hz,1H),4.53(t,J=16.0Hz,2H),4.28-4.22(m,1H),4.08- 4.02(m,1H),3.97(s,3H),3.94-3.88(m,3H),3.62(s,3H),3.25-3.21(m,3 H),3.07(s,3H),3.00-2.94(m,1H),2.87-2.80(m,2H),2.78-2.70(m,3H), 1.95-1.86(m,2H),1.72-1.64(m,1H),1.60-1.53(m,1H).LC-MS(ESI):m / z 866.4[M+H] + .

[0423] Step 2.6. Synthesis of compound XZ1738-C4. Using compounds 13 and 28-C4 as starting materials, the title compound (9.1 mg) was obtained via general step 2. 1H NMR (600MHz, DMSO-d6) δ11.90(s,1H),10.55(s,1H),8.87(d,J=63.0Hz,2H),8.00(d,J=1.9Hz,1H),7.74(s,1H),7.59-7.55(m,2H),7.43 (d,J=4.6Hz,1H),7.25(s,1H),7.14-7.06(m,1H),7.06-7.01(m,1H),6.71-6.65(m,1H),6.32-6.25(m,1H),5.15-5.09(m,1H),4.66-4.63 (m,1H),4.62-4.58(m,1H),4.55-4.50(m,2H),4.27(d,J=7.9Hz,1H),4.03-3.95(m,5H),3.91(t,J=6.7Hz,2H),3.62(s,3H),3.19-3.13(m ,2H),3.09-2.92(m,7H),2.79-2.73(m,2H),2.71-2.64(m,1H),1.93-1.85(m,4H),1.71-1.64(m,1H),1.60-1.53(m,1H).LC-MS(ESI):m / z 880.4[M+H] + .

[0424] Step 2.7. Synthesis of compound XZ1739-C2. Using compounds 13 and 30-C2 as starting materials, the title compound (8.0 mg) was obtained via general step 2. 1H NMR (600MHz, DMSO-d6) δ11.90(d,J=2.9Hz,1H),10.28(s,1H),9.26(s,2H),7.99(d,J=2.3Hz,1H),7.73(s,1H),7.63(s,1H),7.28 (d,J=2.8Hz,1H),7.22-7.18(m,2H),7.10-7.03(m,1H),7.01-6.97(m,2H),6.70-6.64(m,1H),6.32-6.24(m,1H),5.14(d,J=16.6 Hz,1H),4.65(d,J=13.6Hz,1H),4.57(d,J=16.5Hz,1H),4.52(d,J=13.7Hz,1H),4.38(d,J=13.8Hz,1H),4.16(s,2H),4.02-3.97( m,1H),3.74-3.65(m,5H),3.61(s,3H),3.23-3.14(m,5H),3.07(s,3H),2.70(t,J=6.8Hz,2H),1.29-1.15(m,1H).LC-MS(ESI):m / z 799.3[M+H] + .

[0425] Scheme 8. Synthesis of new degrading agents XZ1711, XZ1712, XZ1710, XZ1719 and XZ1723A, XZ1734, XZ1736, XZ1750 and 1751 series

[0426]

[0427] General Procedure 1: Compound 16 (1 equivalent), the corresponding amino compounds 22 / 24 / 20 / 31 / 26 / 28 / 30 (1.2 equivalents), and N,N-diisopropylethylamine (4 equivalents) were dissolved in dimethyl sulfoxide (2 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.5 equivalents) was added, and the reaction was allowed to proceed at room temperature for 30 minutes. After the reaction was monitored by LC-MS until complete, the title compound was obtained by preparative HPLC purification.

[0428] Step 1.1. Synthesis of compound XZ1711-C2. Using compounds 16 and 22-C2 as starting materials, the title compound (13.3 mg) was obtained by general step 1. 1H NMR (600MHz, DMSO-d6) δ11.86(d,J=2.6Hz,1H),11.02(s,1H),8.54(t,J=5.8Hz,1H),8.00(d,J=2.5Hz,1H ),7.88(s,1H),7.69(s,1H),7.64(d,J=8.5Hz,1H),7.60-7.52(m,1H),7.29(d,J=2.3Hz,1H),7.27(s,1H) ,7.22-7.18(m,2H),5.87(s,1H),5.18-4.94(m,2H),4.58(s,1H),4.08(s,2H),3.56(s,7H),3.51-3.39(m ,4H),3.10(s,1H),2.87-2.80(m,4H),2.56-2.45(m,2H),2.00-1.91(m,1H).LC-MS(ESI):m / z882.2[M+H] + .

[0429] Step 1.2. Synthesis of compound XZ1711-C4. Using compounds 16 and 22-C4 as starting materials, the title compound (10.2 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.8Hz,1H),11.09(s,1H),8.43(t,J=5.6Hz,1H),8.07(d,J=2.5Hz,1H),7.92(s,1H),7.76( s,1H),7.71(d,J=8.5Hz,1H),7.65-7.58(m,1H),7.35(d,J=2.3Hz,1H),7.32(s,1H),7.28(d,J=2.7Hz,1H),7.25(dd,J=8.7,2. 4Hz,1H),5.93(s,1H),5.76(s,1H),5.23(s,1H),5.08(dd,J=12.8,5.5Hz,1H),4.63(s,1H),4.27(s,1H),3.65-3.46(m,12H),2 .96-2.84(m,4H),2.63-2.52(m,2H),2.41(t,J=7.4Hz,2H),2.06-1.99(m,1H),1.78-1.70(m,2H).LC-MS(ESI):m / z910.3[M+H] + .

[0430] Step 1.3. Synthesis of compound XZ1711-C6. Using compounds 16 and 22-C6 as starting materials, the title compound (7.0 mg) was obtained by general step 1.1 H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.7Hz,1H),11.09(s,1H),8.42(t,J=5.7Hz,1H),8.07(d,J=2.6Hz,1H),7.92(s,1H),7.76(s,1H ),7.70(d,J=8.5Hz,1H),7.65-7.58(m,1H),7.34(d,J=2.3Hz,1H),7.31-7.26(m,2H),7.24(dd,J=8.7,2.4Hz,1H),5.95(s,1H),5.22 (s,1H),5.08(dd,J=12.8,5.4Hz,1H),4.62(s,1H),4.27(s,1H),3.64-3.59(m,6H),3.55-3.43(m,4H),3.17(s,3H),2.92(s,3H),2. 91-2.84(m,1H),2.63-2.52(m,2H),2.33(t,J=7.5Hz,2H),2.06-1.98(m,1H),1.56-1.45(m,4H),1.34-1.26(m,2H).LC-MS(ESI):m / z 938.3[M+H] + .

[0431] Step 1.4. Synthesis of compound XZ1711-C8. Using compounds 16 and 22-C8 as starting materials, the title compound (8.2 mg) was obtained by general step 1. 1H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.7Hz,1H),11.08(s,1H),8.41(t,J=5.7Hz,1H),8.08(d,J=2.5Hz,1H),7.92(s,1H),7.75(s,1H), 7.70(d,J=8.5Hz,1H),7.66-7.59(m,1H),7.34(d,J=2.3Hz,1H),7.30-7.26(m,2H),7.24(dd,J=8.6,2.3Hz,1H),5.95(s,1H),5.22(s,1 H),5.08(dd,J=12.8,5.5Hz,1H),4.61(s,1H),4.27(s,1H),3.64-3.59(m,7H),3.52-3.46(m,4H),3.19-3.13(m,2H),2.92(s,3H),2.90 -2.84(m,1H),2.63-2.53(m,2H),2.34(t,J=7.5Hz,2H),2.06-2.00(m,1H),1.48(dt,J=20.5,6.9Hz,4H),1.27(s,6H).LC-MS(ESI):m / z 966.3[M+H] + .

[0432] Step 1.5. Synthesis of compound XZ1711-C10. Using compounds 16 and 22-C10 as starting materials, the title compound (9.7 mg) was obtained by general step 1. 1H NMR (600MHz, DMSO-d6) δ11.92(s,1H),11.08(s,1H),8.40(t,J=5.9Hz,1H),8.08(d,J=2.5Hz,1H),7.92(s,1H),7.76(s,1H), 7.70(d,J=8.6Hz,1H),7.65-7.58(m,1H),7.34(s,1H),7.28(s,2H),7.24(d,J=9.1Hz,1H),5.95(s,1H),5.21(s,1H),5.08(d d,J=12.9,5.4Hz,1H),4.61(s,1H),4.27(s,1H),3.64-3.59(m,7H),3.53-3.50(m,2H),3.16(s,1H),2.92(s,3H),2.90-2.84 (m,1H),2.63-2.52(m,2H),2.34(t,J=7.4Hz,2H),2.05-2.00(m,1H),1.54-1.43(m,4H),1.29-1.21(m,10H).LC-MS(ESI):m / z 994.3[M+H] + .

[0433] Step 1.6. Synthesis of compound XZ1711-P2. Using compounds 16 and 22-P2 as starting materials, the title compound (9.7 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.91(s,1H),11.08(s,1H),8.45(t,J=5.7Hz,1H),8.07(d,J=2.4Hz,1H),7.93(s,1H ),7.75(s,1H),7.68(d,J=8.4Hz,1H),7.65-7.58(m,1H),7.33(s,2H),7.27(d,J=2.6Hz,1H),7.21(dd,J=8.9, 2.3Hz,1H),5.94(s,1H),5.18(s,1H),5.08(dd,J=12.9,5.4Hz,1H),4.62(s,1H),4.25(s,1H),3.67-3.57(m,9 H),3.51-3.42(m,8H),2.92(s,3H),2.90-2.85(m,1H),2.63-2.53(m,4H),2.06-2.01(m,1H).LC-MS(ESI):m / z 940.3[M+H] + .

[0434] Step 1.7. Synthesis of compound XZ1711-P4. Using compounds 16 and 22-P4 as starting materials, the title compound (8.2 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.91(d,J=2.7Hz,1H),11.08(s,1H),8.46(t,J=5.7Hz,1H),8.07(d,J=2.5Hz,1H),7.93(s,1 H),7.75(s,1H),7.69(d,J=8.5Hz,1H),7.64-7.58(m,1H),7.33(d,J=2.3Hz,1H),7.30(s,1H),7.27(d,J=2.7Hz,1H),7 .22(dd,J=8.7,2.3Hz,1H),5.95(s,1H),5.18(s,1H),5.08(dd,J=12.8,5.4Hz,1H),4.61(s,1H),4.26(s,1H),3.66-3. 59(m,9H),3.51-3.43(m,16H),2.91(s,3H),2.89-2.84(m,1H),2.63-2.53(m,4H),2.06-1.99(m,1H).LC-MS(ESI):m / z 1028.3[M+H] + .

[0435] Step 1.8. Synthesis of compound XZ1712-C2. Using compounds 16 and 24-C2 as starting materials, the title compound (12.0 mg) was obtained by general step 1. 1H NMR (600MHz, DMSO-d6) δ11.93(s,1H),10.78(s,1H),8.59(t,J=5.9Hz,1H),8.07(d,J=2.5Hz,1H),7.95(s,1H),7.76(s ,1H),7.65-7.59(m,1H),7.34(s,1H),7.28(d,J=2.7Hz,1H),7.09(d,J=8.5Hz,2H),6.94(d,J=8.5Hz,2H),5.96(s,1H), 5.19(s,1H),4.64(s,1H),4.31(s,1H),4.14(s,1H),3.75(dd,J=11.1,5.0Hz,1H),3.65-3.60(m,7H),3.18-3.09(m,4H) ,2.93(s,3H),2.68-2.61(m,2H),2.18-2.12(m,1H),2.04-1.98(m,1H),1.29-1.24(m,1H).LC-MS(ESI):m / z813.2[M+H] + .

[0436] Step 1.9. Synthesis of compound XZ1712-C3. Using compounds 16 and 24-C3 as starting materials, the title compound (12.0 mg) was obtained by general step 1. 1 H NMR (400MHz, DMSO-d6) δ11.92(d,J=2.8Hz,1H),10.78(s,1H),8.41(t,J=5.7Hz,1H),8.06(d,J=2.5Hz,1H),7.92( s,1H),7.76(s,1H),7.64-7.54(m,1H),7.32-7.25(m,2H),7.08(d,J=8.6Hz,2H),6.93(d,J=8.4Hz,2H),5.95(s,1H ),5.18(s,1H),4.63(s,1H),4.25(s,1H),3.74(dd,J=11.1,4.9Hz,1H),3.63(s,3H),3.59-3.55(m,6H),3.18-3.06 (m,4H),2.93(s,3H),2.69-2.57(m,3H),2.49-2.43(m,1H),2.20-2.08(m,1H),2.05-1.95(m,1H).LC-MS(ESI):m / z 827.3[M+H] + .

[0437] Step 1.10. Synthesis of compound XZ1712-C4. Using compounds 16 and 24-C4 as starting materials, the title compound (10.9 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.7Hz,1H),10.78(s,1H),8.43(t,J=5.7Hz,1H),8.07(d,J=2.5Hz,1H),7.93(s,1H),7.76(s,1H),7. 64-7.57(m,1H),7.32(s,1H),7.28(d,J=2.7Hz,1H),7.09(d,J=8.4Hz,2H),6.94(d,J=8.4Hz,2H),5.96(s,1H),5.24(s,1H),4.63(s,1H), 4.27(s,1H),3.75(dd,J=11.2,5.0Hz,1H),3.65-3.60(m,7H),3.32-3.18(m,2H),3.13(t,J=5.2Hz,2H),3.08(t,J=5.3Hz,2H),2.93(s,3H ),2.68-2.60(m,1H),2.49-2.44(m,1H),2.40(t,J=7.3Hz,2H),2.19-2.09(m,1H),2.05-1.97(m,1H),1.77-1.69(m,2H).LC-MS(ESI):m / z 841.3[M+H] + .

[0438] Step 1.11. Synthesis of compound XZ1712-C6. Using compounds 16 and 24-C6 as starting materials, the title compound (10.0 mg) was obtained by general step 1. 1H NMR (600MHz, DMSO-d6) δ11.91(d,J=2.6Hz,1H),10.77(s,1H),8.41(t,J=5.6Hz,1H),8.07(d,J=2.5Hz,1H),7.92(s,1H),7.76( s,1H),7.64-7.58(m,1H),7.29(d,J=11.5Hz,2H),7.08(d,J=8.3Hz,2H),6.92(d,J=8.4Hz,2H),5.95(s,1H),5.21(s,1H),4.62( s,1H),4.27(s,1H),3.75(d,J=4.9Hz,1H),3.63(s,3H),3.58(s,4H),3.21-3.04(m,6H),2.92(s,3H),2.68-2.61(m,1H),2.49-2 .44(m,1H),2.32(t,J=7.5Hz,2H),2.18-2.09(m,1H),2.04-1.98(m,1H),1.54-1.45(m,4H),1.34-1.26(m,2H).LC-MS(ESI):m / z 869.3[M+H] + .

[0439] Step 1.12. Synthesis of compound XZ1712-P2. Using compounds 16 and 24-P2 as starting materials, the title compound (8.5 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.6Hz,1H),10.77(s,1H),8.45(t,J=5.5Hz,1H),8.07(d,J=2.4Hz,1H),7.93(s,1H),7.76 (s,1H),7.61(t,J=10.2Hz,1H),7.32(s,1H),7.28(d,J=2.6Hz,1H),7.07(d,J=8.2Hz,2H),6.91(d,J=8.2Hz,2H),5.95(s,1H), 5.18(s,1H),4.61(s,1H),4.26(s,1H),3.74(dd,J=11.3,5.0Hz,1H),3.65-3.62(m,5H),3.59-3.56(m,4H),3.51-3.46(m,4H) ,3.13-3.05(m,4H),2.91(s,3H),2.67-2.58(m,3H),2.48-2.44(m,1H),2.18-2.09(m,1H),2.04-1.97(m,1H).LC-MS(ESI):m / z 871.3[M+H]+ .

[0440] Step 1.13. Synthesis of compound XZ1712-P4. Using compounds 16 and 24-P4 as starting materials, the title compound (9.7 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.7Hz,1H),10.78(s,1H),8.46(t,J=5.7Hz,1H),8.07(d,J=2.5Hz,1H),7.93(s,1H),7.76 (s,1H),7.65-7.58(m,1H),7.31-7.26(m,2H),7.07(d,J=8.4Hz,2H),6.91(d,J=8.3Hz,2H),5.95(s,1H),5.18(s,1H),4.63(s ,1H),4.26(s,1H),3.74(dd,J=11.1,4.9Hz,1H),3.66-3.57(m,9H),3.49-3.47(m,12H),3.12(t,J=5.1Hz,2H),3.06(t,J=5.2 Hz,2H),2.91(s,3H),2.68-2.57(m,3H),2.49-2.44(m,1H),2.18-2.09(m,1H),2.04-1.97(m,1H).LC-MS(ESI):m / z959.3[M+H] + .

[0441] Step 1.14. Synthesis of compound XZ1719. The title compound (8.0 mg) was obtained from compounds 16 and 20 using general step 1. 1 H NMR(600MHz,DMSO-d6)δ11.94(s,1H),11.08(s,1H),8.10(s,1H),7.99(s,1H),7.78-7.67(m,3 H),7.36(s,1H),7.29(d,J=2.7Hz,1H),7.24(d,J=8.5Hz,1H),7.19(s,1H),6.00(s,1H),5.08( dd,J=12.7,5.5Hz,1H),4.72(s,1H),4.57(s,1H),3.66-3.50(m,5H),3.04(s,4H),2.93-2.85( m,2H),2.63-2.60(m,1H),2.59-2.56(m,1H),2.05-1.96(m,2H),1.24(s,4H).LC-MS(ESI):m / z 825.3[M+H] + .

[0442] Step 1.15. Synthesis of compound XZ1710-C4. Using compounds 16 and 31-C4 as starting materials, the title compound (10.8 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.6Hz,1H),11.09(s,1H),8.46(t,J=5.7Hz,1H),8.05(d,J=2.5Hz,1H),7.92(s,1H ),7.76(s,1H),7.62-7.53(m,2H),7.30-7.26(m,2H),7.10(d,J=8.6Hz,1H),7.01(d,J=7.0Hz,1H),6.54(t,J=6.0Hz,1 H),5.93(s,1H),5.22(s,1H),5.04(dd,J=12.9,5.4Hz,1H),4.62(s,1H),4.27(s,1H),3.62(s,3H),3.30-3.29(m,2H), 2.91(s,3H),2.89-2.83(m,1H),2.64-2.51(m,3H),2.05-1.99(m,1H),1.62-1.52(m,4H),1.24(s,1H).LC-MS(ESI):m / z 827.2[M+H] + .

[0443] Step 1.16. Synthesis of compound XZ1710-C6. Using compounds 16 and 31-C6 as starting materials, the title compound (10.5 mg) was obtained by general step 1. 1H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.8Hz,1H),11.09(s,1H),8.41(t,J=5.6Hz,1H),8.06(d,J=2.5Hz,1H),7.92(s,1H),7.7 6(s,1H),7.64-7.56(m,2H),7.30-7.26(m,2H),7.09(d,J=8.6Hz,1H),7.02(d,J=7.0Hz,1H),6.53(s,1H),5.95(s,1H),5.22 (s,1H),5.05(dd,J=12.8,5.5Hz,1H),4.61(s,1H),4.27(s,1H),3.63(s,3H),3.28(s,2H),3.17(s,2H),2.91(s,3H),2.90-2 .84(m,1H),2.63-2.52(m,2H),2.07-1.99(m,1H),1.59-1.52(m,2H),1.52-1.45(m,2H),1.37-1.30(m,4H).LC-MS(ESI):m / z 855.3[M+H] + .

[0444] Step 1.17. Synthesis of compound XZ1710-C8. Using compounds 16 and 31-C8 as starting materials, the title compound (13.5 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.7Hz,1H),11.09(s,1H),8.40(t,J=5.6Hz,1H),8.07(d,J=2.5Hz,1H),7.91(s,1H),7. 75(s,1H),7.63-7.56(m,2H),7.27(d,J=3.3Hz,2H),7.09(d,J=8.6Hz,1H),7.02(d,J=7.0Hz,1H),6.52(s,1H),5.95(s,1H) ,5.20(s,1H),5.05(dd,J=12.9,5.4Hz,1H),4.60(s,1H),4.26(s,1H),3.63(s,3H),3.15(s,2H),2.91(s,3H),2.89-2.83(m ,1H),2.63-2.52(m,2H),2.06-2.00(m,1H),1.56(t,J=7.3Hz,2H),1.48-1.43(m,2H),1.35-1.24(m,10H).LC-MS(ESI):m / z 883.3[M+H] + .

[0445] Step 1.18. Synthesis of compound XZ1710-P2. Using compounds 16 and 31-P2 as starting materials, the title compound (11.5 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.7Hz,1H),11.09(s,1H),8.50(t,J=5.7Hz,1H),8.06(d,J=2.5Hz,1H),7.93( s,1H),7.76(s,1H),7.63-7.54(m,2H),7.31(s,1H),7.28(d,J=2.7Hz,1H),7.14(d,J=8.6Hz,1H),7.04(d,J=7.0H z,1H),6.61(s,1H),5.95(s,1H),5.18(s,1H),5.05(dd,J=12.9,5.5Hz,1H),4.61(s,1H),4.25(s,1H),3.65-3.58 (m,6H),3.56-3.52(m,5H),2.91(s,3H),2.89-2.82(m,1H),2.62-2.52(m,2H),2.05-1.98(m,1H).LC-MS(ESI):m / z 843.2[M+H] + .

[0446] Step 1.19 Synthesis of compound XZ1710-P3. Using compounds 16 and 31-P3 as starting materials, the title compound (10.7 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.7Hz,1H),11.09(s,1H),8.46(t,J=5.6Hz,1H),8.05(d,J=2.6Hz, 1H),7.92(s,1H),7.75(s,1H),7.62-7.55(m,2H),7.30-7.26(m,2H),7.13(d,J=8.6Hz,1H),7.03(d,J=7 .0Hz,1H),6.59(s,1H),5.94(s,1H),5.28-4.98(m,2H),4.60(s,1H),4.24(s,1H),3.65-3.47(m,13H),2 .95-2.82(m,5H),2.64-2.53(m,2H),2.07-1.98(m,1H),1.37-1.15(m,1H).LC-MS(ESI):m / z887.2[M+H] + .

[0447] Step 1.20. Synthesis of compound XZ1710-P4. Using compounds 16 and 31-P4 as starting materials, the title compound (12.8 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.91(d,J=2.7Hz,1H),11.09(s,1H),8.46(t,J=5.7Hz,1H),8.06(d,J=2.5 Hz,1H),7.92(s,1H),7.75(s,1H),7.64-7.54(m,2H),7.31-7.25(m,2H),7.13(d,J=8.6Hz,1H),7.0 3(d,J=7.0Hz,1H),6.59(s,1H),5.95(s,1H),5.28-5.00(m,2H),4.60(s,1H),4.26(s,1H),3.56-3. 44(m,16H),3.33(s,3H),2.93-2.83(m,4H),2.63-2.52(m,2H),2.06-1.99(m,1H).LC-MS(ESI):m / z 931.3[M+H] + .

[0448] Step 1.21. Synthesis of compound XZ1734-C2. Using compounds 16 and 26-C2 as starting materials, the title compound (8.0 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.93(d,J=2.7Hz,1H),10.35(s,1H),8.59(t,J=5.8Hz,1H),8.08(d,J=2.5Hz,1H) ,7.95(s,1H),7.76(s,1H),7.66-7.60(m,1H),7.45(dd,J=8.5,2.1Hz,1H),7.41(d,J=2.2Hz,1H),7.34(s,1 H),7.28(d,J=2.7Hz,1H),7.19(d,J=8.5Hz,1H),5.95(s,1H),5.18(s,1H),4.65(s,1H),4.28(s,1H),4.12( s,2H),3.86(s,3H),3.65-3.59(m,6H),3.57-3.51(m,7H),2.93(s,3H),2.72-2.67(m,2H).LC-MS(ESI):m / z 872.3[M+H] + .

[0449] Step 1.22. Synthesis of compound XZ1734-C3. Using compounds 16 and 26-C3 as starting materials, the title compound (9.1 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.8Hz,1H),10.35(s,1H),8.40(t,J=5.7Hz,1H),8.06(d,J=2.5Hz,1H ),7.92(s,1H),7.76(s,1H),7.64-7.57(m,1H),7.43(dd,J=8.5,2.2Hz,1H),7.39(d,J=2.2Hz,1H),7.31- 7.26(m,2H),7.18(d,J=8.4Hz,1H),5.95(s,1H),5.18(s,1H),4.63(s,1H),4.26(s,1H),3.86(s,3H),3.6 6-3.57(m,7H),3.45-3.38(m,8H),2.93(s,3H),2.69(t,J=6.7Hz,2H),2.63-2.57(m,2H).LC-MS(ESI):m / z 886.3[M+H] + .

[0450] Step 1.23. Synthesis of compound XZ1734-C4. Using compounds 16 and 26-C4 as starting materials, the title compound (10.0 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.8Hz,1H),10.35(s,1H),8.42(t,J=5.7Hz,1H),8.06(d,J=2.5Hz,1H),7.93(s ,1H),7.76(s,1H),7.64-7.57(m,1H),7.43(dd,J=8.5,2.2Hz,1H),7.39(d,J=2.2Hz,1H),7.31(s,1H),7.28(d,J=2 .7Hz,1H),7.18(d,J=8.5Hz,1H),5.96(s,1H),5.23(s,1H),4.64(s,1H),4.25(s,1H),3.86(s,3H),3.65-3.59(m,1 2H),3.36-3.16(m,3H),2.93(s,3H),2.69(t,J=6.7Hz,2H),2.41-2.36(m,2H),1.76-1.68(m,2H).LC-MS(ESI):m / z 900.3[M+H] + .

[0451] Step 1.24. Synthesis of compound XZ1736-C2. Using compounds 16 and 28-C2 as starting materials, the title compound (8.0 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.93(d,J=2.7Hz,1H),10.55(s,1H),8.56(t,J=5.7Hz,1H),8.08(d,J=2.5Hz,1H),7.96(s,1H),7.76(s,1H) ,7.66-7.61(m,1H),7.58(d,J=8.4Hz,1H),7.46(s,1H),7.35(s,1H),7.29(d,J=2.7Hz,1H),7.05(d,J=8.5Hz,1H),5.95(s,1H),5.20 (s,1H),4.67(s,1H),4.55(d,J=12.8Hz,1H),4.29(s,1H),4.15(s,2H),4.05-4.00(m,1H),3.97(s,3H),3.91(t,J=6.7Hz,2H),3.64( s,3H),3.21-3.16(m,1H),2.95(s,4H),2.80-2.69(m,3H),1.92-1.84(m,2H),1.78-1.68(m,1H),1.65-1.54(m,1H).LC-MS(ESI):m / z 867.3[M+H] + .

[0452] Step 1.25. Synthesis of compound XZ1736-C3. Using compounds 16 and 28-C3 as starting materials, the title compound (7.3 mg) was obtained by general step 1. 1H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.8Hz,1H),10.54(s,1H),8.41(t,J=5.7Hz,1H),8.07(d,J=2.5Hz,1H),7.93(s,1H) ,7.76(s,1H),7.66-7.54(m,2H),7.44(s,1H),7.34-7.26(m,2H),7.05(d,J=8.5Hz,1H),5.95(s,1H),5.20(s,1H),4.80- 4.48(m,2H),4.26(s,1H),3.96(s,4H),3.91(t,J=6.6Hz,2H),3.63(s,3H),3.28-3.07(m,3H),2.96-2.87(m,4H),2.76(t ,J=6.7Hz,2H),2.67-2.56(m,3H),1.88-1.78(m,2H),1.75-1.67(m,1H),1.62-1.49(m,1H).LC-MS(ESI):m / z881.3[M+H] + .

[0453] Step 1.26. Synthesis of compound XZ1736-C4. Using compounds 16 and 28-C4 as starting materials, the title compound (8.7 mg) was obtained by general step 1. 1 H NMR (600MHz, DMSO-d6) δ11.92(d,J=3.0Hz,1H),10.54(s,1H),8.44(t,J=5.7Hz,1H),8.08-8.05(m,1H),7.93(s,1H),7.76(s ,1H),7.64-7.54(m,2H),7.44(s,1H),7.32(s,1H),7.27(d,J=2.8Hz,1H),7.07-7.02(m,1H),5.95(s,1H),5.23(s,1H),4.60( t,J=21.0Hz,2H),4.27(s,1H),4.04-3.85(m,6H),3.63(s,3H),3.57(s,1H),3.19-3.08(m,2H),2.94-2.90(m,4H),2.76(t,J =6.7Hz,2H),2.63-2.58(m,1H),2.43-2.39(m,2H),1.87-1.80(m,2H),1.78-1.62(m,3H),1.59-1.50(m,1H).LC-MS(ESI):m / z 895.4[M+H] + .

[0454] Step 1.27. Synthesis of compound XZ1723A. Using compounds 16 and 30-C2 as starting materials, the title compound (8.2 mg) was obtained via general step 1. 1 H NMR (400MHz, DMSO-d6) δ11.93(d,J=2.8Hz,1H),10.28(s,1H),8.59(t,J=5.8Hz,1H),8.07(d,J=2.6Hz,1H),7.9 5(s,1H),7.76(s,1H),7.67-7.56(m,1H),7.34(s,1H),7.28(d,J=2.7Hz,1H),7.22-7.15(m,2H),7.01-6.95(m, 2H),5.94(s,1H),5.18(s,1H),4.67(s,1H),4.29(s,1H),4.17-4.12(m,1H),3.71(t,J=6.7Hz,2H),3.66-3.61( m,6H),3.20-3.10(m,4H),2.93(s,3H),2.69(t,J=6.7Hz,2H),1.32-1.19(m,1H),-0.06(s,1H).LC-MS(ESI):m / z 814.3[M+H] + .

[0455] Step 1.28. Synthesis of compound XZ1750A. The title compound (5.2 mg) was obtained from compounds 16 and 24-A using the general procedure 1. LC-MS (ESI): m / z 827.3 [M+H] + .

[0456] Step 1.29. Synthesis of compound XZ1750B. The title compound (6.4 mg) was obtained from compounds 16 and 24-B using the general procedure 1. LC-MS (ESI): m / z 827.3 [M+H] + .

[0457] Step 1.30. Synthesis of compound XZ1751A. The title compound (6.2 mg) was obtained from compounds 16 and 24-C using the general procedure 1. LC-MS (ESI): m / z 853.3 [M+H] + .

[0458] Step 1.31. Synthesis of compound XZ1751B. The title compound (5.0 mg) was obtained from compounds 16 and 24-D using the general procedure 1. LC-MS (ESI): m / z 853.3 [M+H] + .

[0459] Scheme 9. Synthesis of the new degradation agents XZ1715, XZ1720A, XZ1735, XZ1718 and XZ1720C series

[0460]

[0461] General Step 1: Compounds 32 / 35 / 37 (1 equivalent), the corresponding linkers 33 or 21 (1.2 equivalents), and N,N-diisopropylethylamine (4 equivalents) were dissolved in dichloromethane (20 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.5 equivalents) was added, and the mixture was reacted at room temperature for 30 minutes. After the reaction was complete as monitored by LC-MS, the mixture was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain the intermediate. This intermediate was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (2 mL) was added. The mixture was reacted at room temperature for 1 hour. After the reaction was complete as monitored by LC-MS, the mixture was concentrated under reduced pressure, and the residue was slurried in diethyl ether to give the corresponding amino compounds 34 / 36 / 38.

[0462] General Step 2: Compound 23 or 35 (1 equivalent), the corresponding linker 39 (1.2 equivalent), and N,N-diisopropylethylamine (2 equivalent) were dissolved in dichloromethane (10 mL) and methanol (2 mL). The mixture was reacted overnight at room temperature, followed by the addition of sodium cyanoborohydride (3 equivalent) and a further reaction time of 1 hour. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain an intermediate. This intermediate was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (2 mL) was added. The mixture was reacted at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure, and the residue was slurried in diethyl ether to give the corresponding amino compound 40 or 42.

[0463] General Step 3: Compound 16 (1 equivalent), the corresponding amino compounds 34 / 36 / 38 / 40 / 42 (1.2 equivalents), and N,N-diisopropylethylamine (4 equivalents) were dissolved in dimethyl sulfoxide (2 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.5 equivalents) were added. The reaction was carried out at room temperature for 30 minutes. After the reaction was monitored by LC-MS until complete, the title compound was obtained by preparative liquid chromatography.

[0464] Step 1.1. Synthesis of Compound 34. Using compounds 32 and 33 as starting materials, the title compound (63 mg) was obtained via general step 1. LC-MS (ESI): m / z 438.2 [M+H] + .

[0465] Step 1.2. Synthesis of Compound 36. Using Compound 35 and Compound 21-C2 as starting materials, the title compound (76 mg) was obtained via general step 1. LC-MS (ESI): m / z 345.2 [M+H] + .

[0466] Step 1.3. Synthesis of compound 38-C2. Using compounds 37 and 21-C2 as starting materials, the title compound (164 mg) was obtained via general step 1. LC-MS (ESI): m / z 331.2 [M+H] + .

[0467] Step 1.4. Synthesis of compound 38-C3. Using compounds 37 and 21-C3 as starting materials, the title compound (132 mg) was obtained via general step 1. LC-MS (ESI): m / z 345.2 [M+H] + .

[0468] Step 1.5. Synthesis of compound 38-C4. Using compounds 37 and 21-C4 as starting materials, the title compound (94 mg) was obtained via general step 1. LC-MS (ESI): m / z 359.2 [M+H] + .

[0469] Step 2.1. Synthesis of compound 40-C2. Using compounds 23 and 39-C2 as starting materials, the title compound (101 mg) was obtained via the general procedure 2. LC-MS (ESI): m / z 317.2 [M+H] + .

[0470] Step 2.2. Synthesis of compound 40-C3. Using compounds 23 and 39-C3 as starting materials, the title compound (84 mg) was obtained via the general procedure 2. LC-MS (ESI): m / z 331.2 [M+H] + .

[0471] Step 2.3. Synthesis of Compound 42. Using compounds 35 and 39-C2 as starting materials, the title compound (91 mg) was obtained via general step 2. LC-MS (ESI): m / z 331.2 [M+H] + .

[0472] Step 3.1. Synthesis of compound XZ1715-P4. Using compounds 16 and 34 as starting materials, the title compound (8.8 mg) was obtained via general step 3. 1H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.7Hz,1H),10.79(s,1H),8.47(t,J=5.7Hz,1H),8.09-8.03(m,2H),7.93(s,1H),7. 76(s,1H),7.64-7.58(m,1H),7.31-7.26(m,2H),7.14(d,J=8.5Hz,2H),6.91(d,J=8.5Hz,2H),5.96(s,1H),5.18(s,1H), 4.59(s,1H),4.46(s,2H),4.26(s,1H),3.79(dd,J=11.6,4.9Hz,1H),3.63(s,3H),3.50-3.47(m,11H),3.31-3.27(m,3H ),2.91(s,3H),2.70-2.62(m,1H),2.50-2.44(m,3H),2.21-2.11(m,1H),2.04-1.97(m,1H).LC-MS(ESI):m / z920.3[M+H] + .

[0473] Step 3.2. Synthesis of compound XZ1720A. Using compounds 16 and 36 as starting materials, the title compound (6.0 mg) was obtained via general step 3. 1H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.7Hz,1H),10.75(d,J=2.2Hz,1H),8.55(dt,J=28.4,5.8Hz,1H),8.07(t,J=2.2Hz,1H),7.93(s,1H),7.76(s ,1H),7.66-7.58(m,1H),7.33(d,J=3.0Hz,1H),7.28(d,J=2.7Hz,1H),7. 01(t,J=7.8Hz,2H),6.72(t,J=9.5Hz,2H),5.94(s,1H),5.17(s,1H),4.6 3(s,1H),4.28(s,1H),4.08(s,1H),3.99(s,1H),3.69-3.63(m,6H),3.57 -3.49(m,3H),3.45-3.42(m,1H),2.90(d,J=7.2Hz,3H),2.66-2.57(m,1H ),2.48-2.44(m,1H),2.15-2.07(m,1H),2.04-1.99(m,1H),1.93-1.88(m ,1H),1.85-1.80(m,1H),1.30-1.22(m,2H).LC-MS(ESI):m / z827.3[M+H] + .

[0474] Step 3.3. Synthesis of compound XZ1735-C2. Using compounds 16 and 38-C2 as starting materials, the title compound (9.1 mg) was obtained via general step 3. 1H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.8Hz,1H),10.80(s,1H),8.58(t,J=5.8Hz,1H),8.07(d,J=2.5Hz,1H),7.95(s,1H),7. 76(s,1H),7.65-7.58(m,1H),7.35(s,1H),7.28(d,J=2.7Hz,1H),7.20(t,J=7.9Hz,1H),6.90-6.84(m,2H),6.68(d,J=7.5H z,1H),5.95(s,1H),5.18(s,1H),4.66(s,1H),4.28(s,1H),4.15(s,2H),3.79(dd,J=11.4,4.9Hz,1H),3.65-3.60(m,7H),3 .21-3.10(m,4H),2.93(s,3H),2.69-2.62(m,1H),2.49-2.46(m,1H),2.26-2.17(m,1H),2.06-2.00(m,1H).LC-MS(ESI):m / z 813.3[M+H] + .

[0475] Step 3.4. Synthesis of compound XZ1735-C3. Using compounds 16 and 38-C3 as starting materials, the title compound (10.0 mg) was obtained by general step 3. 1 H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.7Hz,1H),10.80(s,1H),8.41(t,J=5.7Hz,1H),8.06(d,J=2.5Hz,1H),7.92(s,1H), 7.76(s,1H),7.63-7.56(m,1H),7.32-7.26(m,2H),7.19(t,J=7.8Hz,1H),6.88-6.83(m,2H),6.67(d,J=7.5Hz,1H),5.95 (s,1H),5.19(s,1H),4.63(s,1H),4.25(s,1H),3.78(dd,J=11.3,4.9Hz,1H),3.64-3.56(m,9H),3.43-3.40(m,2H),3.17 -3.14(m,2H),3.12-3.07(m,2H),2.93(s,3H),2.66-2.62(m,2H),2.23-2.18(m,1H),2.05-1.98(m,1H).LC-MS(ESI):m / z 827.3[M+H] + .

[0476] Step 3.5. Synthesis of compound XZ1735-C4. Using compounds 16 and 38-C4 as starting materials, the title compound (9.7 mg) was obtained via general step 3. 1 H NMR (600MHz, DMSO-d6) δ11.92(d,J=2.7Hz,1H),10.80(s,1H),8.42(t,J=5.7Hz,1H),8.07(d,J=2.5Hz,1H),7.93(s,1H),7.76(s,1H),7. 61(ddd,J=11.3,8.2,2.5Hz,1H),7.32(s,1H),7.28(d,J=2.7Hz,1H),7.19(t,J=7.8Hz,1H),6.89-6.83(m,2H),6.67(d,J=7.5Hz,1H),5.9 6(s,1H),5.24(s,1H),4.63(s,1H),4.27(s,1H),3.78(dd,J=11.3,5.0Hz,1H),3.64-3.60(m,5H),3.36-3.17(m,3H),3.16-3.07(m,4H),2 .93(s,3H),2.69-2.62(m,1H),2.43-2.37(m,2H),2.25-2.15(m,1H),2.09(s,2H),2.07-1.99(m,1H),1.78-1.70(m,2H).LC-MS(ESI):m / z 841.3[M+H] + .

[0477] Step 3.6. Synthesis of compound XZ1718-C2. Using compounds 16 and 40-C2 as starting materials, the title compound (9.0 mg) was obtained via general step 3. 1H NMR (600MHz, DMSO-d6) δ11.95(s,1H),10.79(s,1H),9.15(s,1H),8.70(s,1H),8.08(d,J=2.9Hz,1H),7.98(s,1H),7.78(s,1H) ,7.64(t,J=10.4Hz,1H),7.39(s,1H),7.29(d,J=2.7Hz,1H),7.12(d,J=8.2Hz,2H),6.98(d,J=8.2Hz,2H),5.95(s,1H),5.25(s, 1H),4.72(s,1H),4.28(s,1H),3.84(d,J=13.1Hz,2H),3.77(dd,J=11.3,5.0Hz,1H),3.70-3.62(m,6H),3.27-3.20(m,4H),3.0 0-2.98(m,4H),2.68-2.63(m,1H),2.48-2.45(m,2H),2.18-2.13(m,1H),2.04-1.97(m,1H),1.26-1.23(m,1H).LC-MS(ESI):m / z 799.3[M+H] + .

[0478] Step 3.7. Synthesis of compound XZ1718-C3. Using compounds 16 and 40-C3 as starting materials, the title compound (8.0 mg) was obtained via general step 3. 1H NMR (600MHz, DMSO-d6) δ11.94(d,J=2.8Hz,1H),10.79(s,1H),9.43(s,1H),8.56(t,J=5.9Hz,1H),8.07(d,J=2.5Hz,1H),7.95(s,1H), 7.78(s,1H),7.66-7.59(m,1H),7.34(s,1H),7.29(d,J=2.7Hz,1H),7.12(d,J=8.5Hz,2H),6.98(d,J=8.7Hz,2H),5.96(s,1H),5.26(s ,1H),4.71(s,1H),4.27(s,1H),3.83(d,J=12.9Hz,2H),3.76(dd,J=11.4,5.0Hz,1H),3.63(s,3H),3.58-3.55(m,2H),3.22-3.12(m,5 H),2.99-2.97(m,4H),2.68-2.62(m,1H),2.48-2.45(m,1H),2.18-2.12(m,1H),2.04-1.84(m,4H),1.26-1.23(m,1H).LC-MS(ESI):m / z 813.4[M+H] + .

[0479] Step 3.8. Synthesis of compound XZ1720C. Using compounds 16 and 42 as starting materials, the title compound (10.0 mg) was obtained by general step 3. 1 H NMR (600MHz, DMSO-d6) δ11.95(d,J=2.7Hz,1H),10.76(s,1H),9.15(s,1H),8.67(s,1H),8.08(d,J=2.5Hz,1H),7.97(s,1 H),7.78(s,1H),7.67-7.60(m,1H),7.36(s,1H),7.29(d,J=2.7Hz,1H),7.05(d,J=8.6Hz,2H),6.74(d,J=8.8Hz,2H),5.9 4(s,1H),5.23(s,1H),4.71(s,1H),4.27(s,1H),3.78(d,J=16.2Hz,1H),3.71(dd,J=11.1,4.9Hz,2H),3.64(s,11H),2.9 8(s,3H),2.68-2.60(m,1H),2.48-2.44(m,2H),2.19-2.08(m,3H),2.02-1.96(m,1H),1.33-1.23(m,1H).LC-MS(ESI):m / z 813.3[M+H]+ .

[0480] Scheme 10. Synthesis of the new degradation agents XZ1721A, XZ1721B, XZ1724, XZ1725, XZ1726, XZ1728, XZ1731, XZ1732, XZ1727 and XZ1755 series

[0481]

[0482]

[0483] General Step 1: Compound 43 (1 equivalent), the corresponding heterocycle 44 (2 equivalents), palladium acetate (0.1 equivalent), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (0.1 equivalent), and cesium carbonate (3 equivalents) were added to 1,4-dioxane (10 mL) and reacted overnight at 90 °C under nitrogen protection. After the reaction was complete as monitored by TLC, the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain the coupling compound. This coupling compound and 10% palladium on carbon (20 mg) were added to methanol (10 mL) and reacted overnight at room temperature under hydrogen atmosphere. After the reaction was complete as monitored by LC-MS, the mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the reduction product. The reduction product was dissolved in dichloromethane (10 mL) and trifluoroacetic acid (2 mL) was added. The mixture was reacted at room temperature for 1 hour. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue was then slurried in diethyl ether to obtain 45a-g of the corresponding amino compound.

[0484] General Step 2: Compound 43 (1 equivalent), boron ester 46 (2 equivalents), tetra-triphenylphosphine palladium (0.1 equivalents), and potassium carbonate (3 equivalents) were added to 1,4-dioxane (10 mL) and water (2 mL), and reacted overnight at 80 °C under nitrogen protection. After the reaction was complete as monitored by TLC, the mixture was extracted with dichloromethane, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain the coupling product. This coupling product and 10% palladium on carbon (20 mg) were added to methanol (10 mL), and the mixture was reacted overnight at room temperature under hydrogen atmosphere. After the reaction was complete as monitored by LC-MS, the mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the reduced product. The reduced product was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (2 mL) was added. The mixture was reacted at room temperature for 1 hour. After the reaction was complete as monitored by LC-MS, the mixture was concentrated under reduced pressure, and the residue was slurried in diethyl ether to obtain the corresponding amino compound 47.

[0485] General Step 3: Compound 16 (1 equivalent), the corresponding amino compound 45 or 47 (1.2 equivalents), and N,N-diisopropylethylamine (4 equivalents) were dissolved in dimethyl sulfoxide (2 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.5 equivalents) were added. The mixture was reacted at room temperature for 30 minutes. After the reaction was monitored by LC-MS until complete, the title compound was obtained by preparative liquid chromatography.

[0486] General Step 4: Compound 43B (1 equivalent), the corresponding amine compound 44 (2 equivalents), palladium acetate (0.1 equivalents), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (0.1 equivalents), and cesium carbonate (3 equivalents) were added to 1,4-dioxane (10 mL) and reacted overnight at 90 °C under nitrogen protection. After the reaction was complete as monitored by TLC, the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain the coupling intermediate. This compound was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (2 mL) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was complete as monitored by LC-MS, the mixture was concentrated under reduced pressure, and the residue was slurried in diethyl ether to obtain the corresponding amino intermediate. The intermediate (1 equivalent), linker 21-C2 (1.2 equivalent), and N,N-diisopropylethylamine (4 equivalent) were dissolved in dichloromethane (20 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.5 equivalent) was added. The reaction was carried out at room temperature for 30 minutes. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue was purified by column chromatography to obtain a higher intermediate, which was dissolved in dichloromethane (10 mL) and trifluoroacetic acid (2 mL) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue was slurried in diethyl ether to give the corresponding amino compound 45 h-j.

[0487] Step 1.1. Synthesis of compound 45a-C1. Using compounds 43 and 44a-C1 as starting materials, the title compound (55 mg) was obtained via general step 1. LC-MS (ESI): m / z 330.2 [M+H] + .

[0488] Step 1.2. Synthesis of compound 45a-C2. Using compounds 43 and 44a-C2 as starting materials, the title compound (62 mg) was obtained via general step 1. LC-MS (ESI): m / z 316.2 [M+H] + .

[0489] Step 1.3. Synthesis of compound 45b. Using compounds 43 and 44b as starting materials, the title compound (109 mg) was obtained via general step 1. LC-MS (ESI): m / z 328.2 [M+H] + .

[0490] Step 1.4. Synthesis of compound 45c. Using compounds 43 and 44c as starting materials, the title compound (63 mg) was obtained via general step 1. LC-MS (ESI): m / z 342.2 [M+H] + .

[0491] Step 1.5. Synthesis of compound 45d. Using compounds 43 and 44d as starting materials, the title compound (78 mg) was obtained via general step 1. LC-MS (ESI): m / z 314.2 [M+H] + .

[0492] Step 1.6. Synthesis of compound 45e. Using compounds 43 and 44e as starting materials, the title compound (90 mg) was obtained via general step 1. LC-MS (ESI): m / z 344.2 [M+H] + .

[0493] Step 1.7. Synthesis of compound 45f. Using compounds 43 and 44f as starting materials, the title compound (84 mg) was obtained via general step 1. LC-MS (ESI): m / z 342.2 [M+H] + .

[0494] Step 1.8. Synthesis of Compound 45g. Using compounds 43 and 44g as starting materials, the title compound (75mg) was obtained via general step 1. LC-MS (ESI): m / z 328.2 [M+H] + .

[0495] Step 2.1. Synthesis of Compound 47. Using compounds 43 and 46 as starting materials, the title compound (65 mg) was obtained via the general procedure 2. LC-MS (ESI): m / z 330.2 [M+H] + .

[0496] Step 3.1. Synthesis of compound XZ1721A. Using compounds 16 and 45a-C1 as starting materials, the title compound (10.0 mg) was obtained via general step 3. 1H NMR (600MHz, DMSO-d6) δ11.93(d,J=2.7Hz,1H),10.82(s,1H),8.43(t,J=5.6Hz,1H),8.08(d,J=2.5Hz,1H),7. 94(s,1H),7.76(s,1H),7.66-7.59(m,1H),7.32-7.09(m,6H),5.98(s,1H),5.27(s,1H),4.64(s,1H),4.27(s, 1H),3.83(s,1H),3.68-3.57(m,7H),2.94(s,3H),2.67-2.62(m,1H),2.50-2.47(m,5H),2.20-2.15(m,1H),2. 06-1.99(m,1H),1.87-1.84(m,1H),1.62-1.55(m,1H),1.51-1.46(m,2H),1.38-1.34(m,1H).LC-MS(ESI):m / z 798.4[M+H] + .

[0497] Step 3.2. Synthesis of compound XZ1721B. Using compounds 16 and 45a-C2 as starting materials, the title compound (9.0 mg) was obtained via general step 3. 1 H NMR (600MHz, DMSO-d6) δ11.93(d,J=2.7Hz,1H),10.81(s,1H),8.54(s,1H),8.08(d,J=2.5Hz,1H),7.94( s,1H),7.77(s,1H),7.66-7.60(m,1H),7.37-7.01(m,6H),5.95(s,1H),5.19(s,1H),4.61(s,1H),4.29( s,1H),3.84-3.78(m,1H),3.67-3.60(m,6H),3.19-3.08(m,3H),2.94(s,3H),2.70-2.62(m,1H),2.48-2 .43(m,2H),2.19-2.14(m,1H),2.05-1.98(m,1H),1.86-1.79(m,2H),1.42-1.29(m,2H).LC-MS(ESI):m / z 784.3[M+H] + .

[0498] Step 3.3. Synthesis of compound XZ1724. Using compounds 16 and 45b as starting materials, the title compound (12.0 mg) was obtained via general step 3. 1H NMR (600MHz, DMSO-d6) δ11.93(s,1H),10.80(s,1H),8.65(d,J=7.3Hz,1H),8.06(d,J=2.4Hz,1H),7 .93(s,1H),7.76(s,1H),7.65-7.58(m,1H),7.36-6.93(m,6H),5.94(s,1H),5.23(s,1H),4.59(s,1 H),4.38-4.21(m,2H),3.80-3.74(m,1H),3.63(s,3H),3.23-3.11(m,4H),2.92(s,3H),2.66-2.62( m,1H),2.26-2.12(m,3H),2.04-1.98(m,1H),1.88-1.64(m,6H),1.27-1.23(m,1H).LC-MS(ESI):m / z 810.4[M+H] + .

[0499] Step 3.4. Synthesis of compound XZ1725. Using compounds 16 and 45c as starting materials, the title compound (11.0 mg) was obtained via general step 3. 1 H NMR(600MHz,DMSO-d6)δ11.93(d,J=2.9Hz,1H),10.80(s,1H),8.12(s,1H),7.97(s,1H),7. 71(s,2H),7.32-7.01(m,6H),5.99(s,1H),4.68(s,1H),4.52(s,1H),4.22(s,1H),3.90-3. 73(m,2H),3.64(s,3H),3.26-3.16(m,6H),3.02(s,3H),2.68-2.62(m,1H),2.55(s,1H),2. 20-2.14(m,1H),2.05-1.97(m,1H),1.77-1.42(m,8H),1.29-1.20(m,1H).LC-MS(ESI):m / z 824.4[M+H] + .

[0500] Step 3.5. Synthesis of compound XZ1726. Using compounds 16 and 45d as starting materials, the title compound (8.0 mg) was obtained via general step 3. 1H NMR (600MHz, DMSO-d6) δ11.93(d,J=2.6Hz,1H),10.78(s,1H),8.09(d,J=2.4Hz,1H),7.97(s,1H),7.7 5(s,1H),7.70-7.63(m,1H),7.28(d,J=2.7Hz,1H),7.19(s,1H),7.15-6.95(m,4H),5.94(s,1H),4.91 -4.72(m,2H),4.26(s,1H),3.78-3.71(m,4H),3.64(s,3H),3.21-3.11(m,4H),3.03(s,3H),2.68-2.6 1(m,1H),2.18-2.10(m,1H),2.04-1.96(m,2H),1.86-1.81(m,3H),1.26-1.23(m,2H).LC-MS(ESI):m / z 796.3[M+H] + .

[0501] Step 3.6. Synthesis of compound XZ1728. Using compounds 16 and 45e as starting materials, the title compound (9.0 mg) was obtained via general step 3. 1 H NMR (600MHz, DMSO-d6) δ11.92(s,1H),10.77(s,1H),8.10(s,1H),7.97(s,1H),7.71(s,1H),7.61(d,J= 12.0Hz,1H),7.28(d,J=2.6Hz,1H),7.13-7.05(m,3H),6.88(d,J=15.2Hz,2H),6.02(s,1H),4.66(s,1H ),4.53(s,1H),4.15(s,1H),3.80-3.72(m,3H),3.64(s,3H),3.14-2.88(m,11H),2.66-2.61(m,1H),2. 47-2.44(m,1H),2.17-2.11(m,1H),2.03-1.94(m,2H),1.80(s,1H),1.69-1.55(m,2H).LC-MS(ESI):m / z 826.3[M+H] + .

[0502] Step 3.7. Synthesis of compound XZ1731. Using compounds 16 and 45f as starting materials, the title compound (8.0 mg) was obtained via general step 3. 1H NMR(600MHz,DMSO-d6)δ11.92(s,1H),10.77(s,1H),8.11(s,1H),7.96(s,1H),7.72 -7.57(m,2H),7.28(d,J=2.7Hz,1H),7.14-6.88(m,5H),6.01(s,1H),4.67(s,1H),4. 52(s,1H),4.23(s,1H),3.78-3.69(m,2H),3.64(s,3H),3.19-2.85(m,10H),2.66-2. 61(m,1H),2.15-2.09(m,1H),2.05-1.98(m,1H),1.71-1.22(m,9H).LC-MS(ESI):m / z 824.4[M+H] + .

[0503] Step 3.8. Synthesis of compound XZ1732. Using compound 16 and compound 45 g as starting materials, the title compound (10.0 mg) was obtained by general step 3. 1 H NMR (600MHz, DMSO-d6) δ11.92(s,1H),10.78(d,J=18.0Hz,1H),8.08(dd,J=10.4,2.5Hz,1H),7.94(d,J=13.5Hz,1H),7.71(d,J= 17.0Hz,1H),7.52(t,J=10.3Hz,1H),7.30-7.25(m,2H),7.20(s,1H),7.17-7.02(m,3H),5.97(s,1H),4.68(d,J=15.9Hz,2H),4.2 7(s,1H),3.84-3.71(m,1H),3.63(d,J=6.1Hz,3H),3.53-3.49(m,2H),3.17(s,1H),3.04-2.92(m,5H),2.64(d,J=17.5Hz,1H),2 .48-2.46(m,2H),2.14(dd,J=23.2,11.4Hz,1H),2.05-1.96(m,1H),1.80-1.56(m,7H),1.24(s,1H).LC-MS(ESI):m / z810.4[M+H] + .

[0504] Step 3.9. Synthesis of compound XZ1727. Using compounds 16 and 47 as starting materials, the title compound (8.0 mg) was obtained via general step 3. 1H NMR (600MHz, DMSO-d6) δ11.93(d,J=2.7Hz,1H),10.82(s,1H),8.54(t,J=5.7Hz,1H),8.07(d,J=2.5Hz,1H),7.95(s,1H),7.76(s,1H),7.62(t,J=1 0.4Hz,1H),7.34(s,1H),7.28(d,J=2.7Hz,1H),7.21(d,J=7.9Hz,2H),7. 16(d,J=7.9Hz,2H),5.95(s,1H),5.20(s,1H),4.65(s,1H),4.51(d,J=12 .8Hz,1H),4.29(s,1H),4.12(s,2H),3.98(d,J=13.4Hz,1H),3.82(dd,J= 11.5,4.9Hz,1H),3.64(s,3H),3.19-3.09(m,2H),2.94(s,3H),2.82-2.7 4(m,1H),2.70-2.61(m,2H),2.22-2.14(m,1H),2.06-2.01(m,1H),1.81( s,2H),1.66-1.57(m,1H),1.49-1.44(m,1H).LC-MS(ESI):m / z812.3[M+H] + .

[0505] Step 3.10. Synthesis of compound XZ1755A. Using compounds 16 and 44h as starting materials, the title compound (7.1 mg) was obtained via the general procedure 3. LC-MS (ESI): m / z 840.3 [M+H] + .

[0506] Step 3.11. Synthesis of compound XZ1755B. Using compounds 16 and 44i as starting materials, the title compound (5.6 mg) was obtained via general step 3. LC-MS (ESI): m / z 828.3 [M+H] + .

[0507] Step 3.12. Synthesis of compound XZ1755C. Using compounds 16 and 44j as starting materials, the title compound (3.9 mg) was obtained via general step 3. m / z 828.3 [M+H] + .

[0508] Step 4.1. Synthesis of compound 45h. Using compounds 43B and 44h as starting materials, the title compound (75 mg) was obtained via the general procedure 4. LC-MS (ESI): m / z 358.2 [M+H] + .

[0509] Step 4.2. Synthesis of Compound 45i. Using compounds 43B and 44i as starting materials, the title compound (82 mg) was obtained via the general procedure 4. LC-MS (ESI): m / z 346.2 [M+H] + .

[0510] Step 4.3. Synthesis of compound 45j. Using compounds 43B and 44j as starting materials, the title compound (61 mg) was obtained via general step 4. LC-MS (ESI): m / z 346.2 [M+H] + .

[0511] Scheme 11. Synthesis of new degradation agents XZ1730, XZ1717, XZ1740 and XZ1723B series

[0512]

[0513] General Step 1: Compounds 23 (1 equivalent), 48 (1.2 equivalent), and N,N-diisopropylethylamine (4 equivalent) were dissolved in dichloromethane (20 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.5 equivalent) was added. The reaction was carried out at room temperature for 30 minutes. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain an intermediate. This intermediate was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (2 mL) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure, and the residue was slurried in diethyl ether to obtain amino compound 49.

[0514] General Step 2: Compound 17 (1 equivalent), the corresponding amino compound 49 / 24-C2 / 45a-C2 / 30 (1.2 equivalents), and N,N-diisopropylethylamine (2 equivalents) were dissolved in dichloromethane (2 mL) and methanol (1 mL). The mixture was reacted overnight at room temperature, followed by the addition of sodium cyanoborohydride (3 equivalents) and a further reaction time of 1 hour. After the reaction was completed by LC-MS monitoring, the mixture was concentrated under reduced pressure, and the residue was purified by preparative liquid chromatography to obtain the title compound.

[0515] Step 1.1. Synthesis of compound 38. Using compounds 23 and 48 as starting materials, the title compound (151 mg) was obtained by general step 1.

[0516] Step 2.1. Synthesis of compound XZ1730. Using compounds 17 and 49 as starting materials, the title compound (9.3 mg) was obtained via general step 2. 1H NMR(600MHz,DMSO-d6)δ11.95(s,1H),10.79(s,1H),9.44(s,1H),8.13(s,1H),7.97(s,1H),7.74(s,1 H),7.64(t,J=10.4Hz,1H),7.49(s,1H),7.29(d,J=2.2Hz,1H),7.10(d,J=8.2Hz,2H),6.94(d,J=8.0Hz ,2H),6.00(s,1H),4.69(s,2H),4.44(s,1H),4.34(s,1H),3.78-3.73(m,2H),3.66-3.60(m,6H),3.20 -3.10(m,12H),2.74-2.71(m,2H),2.17-2.12(m,1H),2.05-1.98(m,1H),1.24(s,1H).LC-MS(ESI):m / z 813.3[M+H] + .

[0517] Step 2.1. Synthesis of compound XZ1717-C2. Using compounds 17 and 24-C2 as starting materials, the title compound (16.6 mg) was obtained by general step 2. 1 H NMR (600MHz, DMSO-d6) δ11.93(d,J=2.6Hz,1H),10.78(s,1H),9.01(s,2H),8.12(d,J=2.5Hz,1H),7.93(s,1H),7.71(s ,1H),7.69-7.64(m,1H),7.44(s,1H),7.28(d,J=2.6Hz,1H),7.10(d,J=8.3Hz,2H),6.94(d,J=8.4Hz,2H),6.06-5.85(m ,1H),4.92-4.66(m,2H),4.32-4.06(m,5H),3.75(dd,J=11.3,4.9Hz,1H),3.68-3.65(m,2H),3.64(s,3H),3.52-3.48( m,2H),3.20-3.10(m,7H),2.69-2.61(m,1H),2.47-2.44(m,1H),2.17-2.10(m,1H),2.04-1.98(m,1H).LC-MS(ESI):m / z 799.3[M+H] + .

[0518] Step 2.3. Synthesis of compound XZ1740-C2. Using compounds 17 and 45a as starting materials, the title compound (15.3 mg) was obtained via general step 2. 1H NMR (400MHz, DMSO-d6) δ11.95(d,J=2.7Hz,1H),10.79(s,1H),8.70-8.40(m,2H),8.12(d,J=2.5Hz,1H),7 .94(s,1H),7.71(s,1H),7.71-7.61(m,1H),7.42(s,1H),7.28(d,J=2.7Hz,1H),7.15-6.96(m,4H),5.98(s ,1H),4.80(s,2H),4.27(s,2H),3.80-3.67(m,9H),3.12(s,3H),2.99(s,2H),2.76(s,1H),2.71-2.58(m,1 H),2.22-1.95(m,2H),1.74(d,J=11.8Hz,2H),1.64-1.44(m,3H),1.31(d,J=11.9Hz,2H).LC-MS(ESI):m / z 784.3[M+H] + .

[0519] Step 2.2. Synthesis of compound XZ1723B. Using compounds 17 and 30 as starting materials, the title compound (11.2 mg) was obtained by general step 2. 1 H NMR (600MHz, DMSO-d6) δ11.95(s,1H),10.30(s,1H),9.03(d,J=46.3Hz,2H),8.12(s,1H),7.93(s,1 H),7.73-7.65(m,2H),7.44(s,1H),7.28(s,1H),7.20(d,J=8.0Hz,2H),6.99(d,J=8.3Hz,2H),5.96( s,1H),4.80(d,J=68.7Hz,2H),4.20(d,J=46.6Hz,4H),3.73-3.65(m,4H),3.65-3.62(m,3H),3.52- 3.48(m,3H),3.21-3.18(m,2H),3.15(s,2H),3.12-3.10(m,3H),2.72-2.67(m,2H).LC-MS(ESI):m / z 800.3[M+H] + .

[0520] Scheme 12. Synthesis of the new degradation agent XZ1757 series

[0521]

[0522] General Step 1: Dissolve compound I-1 / I-1' (1.0 equivalent), potassium carbonate (2.5 equivalent), 1-imidazolium sulfonyl azide hydrochloride (1.1 equivalent), and anhydrous copper sulfate (0.1 equivalent) in anhydrous methanol (20 mL) and react overnight at room temperature under nitrogen protection. After the reaction is complete as monitored by LC-MS, concentrate under reduced pressure, and purify the residue by column chromatography (methanol / dichloromethane = 0-10%) to give the corresponding intermediate I-1B / I-1B'.

[0523] Step 1.1. Synthesis of compound I-1B. Using compound I-1 as the starting material, the title compound (570 mg) was obtained by general step 1.

[0524] Step 1.2. Synthesis of compound I-1B'. Using compound I-1' as the starting material, the title compound (318 mg) was obtained by general step 1.

[0525] Step 2: Compound 16 (1.0 equivalent), 11-dodecyn-1-amine (1.2 equivalent), and N,N-diisopropylethylamine (4 equivalent) were dissolved in dichloromethane (5 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.05 equivalent) was added. The mixture was reacted at room temperature for 1 hour. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue was purified by column chromatography (methanol / dichloromethane = 0-10%) to give compound 16A (118 mg).

[0526] Step 3: Compound 13 (1 equivalent), 11-dodecyn-1-amine (1.2 equivalents), and N,N-diisopropylethylamine (2 equivalents) were dissolved in dichloromethane (2 mL) and methanol (1 mL). The mixture was reacted overnight at room temperature, and then sodium cyanoborohydride (3.0 equivalents) was added to continue the reaction for 1 hour. After the reaction was completed by LC-MS monitoring, the mixture was concentrated under reduced pressure, and the residue was purified by preparative liquid chromatography to obtain compound 13A (61 mg).

[0527] General Step 4: Compounds 16A / 13A (1.0 equivalent), I-1B / I-1B' / I-1C / I-1C' / I-1D / I-1D' (1.0 equivalent), copper sulfate pentahydrate (0.1 equivalent), and sodium ascorbate (0.2 equivalent) were dissolved in 1,4-dioxane (3 mL), tert-butanol (2 mL), and water (1 mL). The reaction was carried out at 50 °C for 1 hour under nitrogen protection. After the reaction was completed as monitored by LC-MS, the title compound was obtained by preparative liquid chromatography.

[0528] Step 3.1. Synthesis of compound XZ1757A. Using compounds 16A and I-1B as starting materials, the title compound (8 mg) was obtained via general step 4. LC-MS (ESI): m / z 1136.5 [M+H] + .

[0529] Step 3.2. Synthesis of compound XZ1757B. Using compounds 16A and I-1B' as starting materials, the title compound (6 mg) was obtained via general step 4. LC-MS (ESI): m / z 1122.4 [M+H] + .

[0530] Step 3.3. Synthesis of compound XZ1757C. Using compounds 13A and I-1C as starting materials, the title compound (10 mg) was obtained via general step 4. LC-MS (ESI): m / z 1105.5 [M+H] + .

[0531] Step 3.4. Synthesis of compound XZ1757C'. Using compounds 13A and I-1C' as starting materials, the title compound (5 mg) was obtained via general step 4. LC-MS (ESI): m / z 1091.5 [M+H] + .

[0532] Step 3.5. Synthesis of compound XZ1757D. Using compounds 13A and I-1D as starting materials, the title compound (7 mg) was obtained via general step 4. LC-MS (ESI): m / z 1119.5 [M+H] + .

[0533] Step 3.6. Synthesis of compound XZ1757D'. Using compounds 13A and I-1D' as starting materials, the title compound (7 mg) was obtained via general step 4. LC-MS (ESI): m / z 1105.5 [M+H] + .

[0534] Example 2. Synthesis of novel degradation agent conjugates

[0535] Scheme 1. Synthesis of GNE-987-OH, XZ1509, XZ1512, XZ1510 and XZ1513

[0536]

[0537] Step 1. Synthesis of Compound I-3. Compound I-1 (1.50 g, 2.68 mmol), compound I-2 (669 mg, 2.94 mmol), and N,N-diisopropylethylamine (1.77 mL, 10.7 mmol) were dissolved in dichloromethane (60 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.02 g, 2.68 mmol) was added, and the reaction was carried out at room temperature for 1 hour. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue was purified by column chromatography (methanol / dichloromethane = 0-10%) to give the title compound I-3 (1.40 g, 80% yield). 1 H NMR (400MHz, DMSO-d6) δ9.81(s,1H),8.96(s,1H),8.48(t,J=6.1Hz,1H),7.86(d,J=9.4Hz,1H),7.38(d,J=7.8Hz,1H),6.91(d,J =1.8Hz,1H),6.80(dd,J=7.8,1.8Hz,1H),5.13(d,J=3.6Hz,1H),4.54(d,J=9.4Hz,1H),4.45(t,J=8.0Hz,1H),4.35(s,1H),4.29( dd,J=16.2,6.5Hz,1H),4.12(dd,J=16.2,5.5Hz,1H),3.69-3.61(m,2H),3.33-3.29(m,2H),2.45(s,3H),2.32-2.22(m,1H),2.1 4-2.07(m,1H),2.07-2.00(m,1H),1.97-1.87(m,1H),1.56-1.44(m,4H),1.28-1.22(m,12H),0.93(s,9H).HRMS(ESI-TOF):Calcd for C 33 H 50 N7O5S, [M+H] + ,656.3594,found 656.3592.

[0538] Step 2. Synthesis of compounds I-8a and I-8b:

[0539] Step 2.1. Synthesis of compound I-8a. Compound I-7a (2.00 g, 5.08 mmol) and N,N-diisopropylethylamine (1.68 mL, 10.2 mmol) were dissolved in acetonitrile (40 mL), and ethylsulfonyl chloride (578 μL, 6.10 mmol) was added. The reaction was carried out at room temperature for 2 hours. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue was purified by column chromatography (ethyl acetate / dichloromethane = 0-40%) to give the title compound I-8a (1.47 g, 70% yield). 1 H NMR (400MHz, DMSO-d6) δ10.09(s,1H),8.10(d,J=6.9Hz,1H),7.59(d,J=8.2Hz,2H),7.37(d,J=8.3Hz,2H),6.73(d,J=8.9Hz,1H),4.72(s,2 H),4.49-4.36(m,1H),3.88-3.78(m,1H),1.99-1.90(m,1H),1.39(s,9H),1.31(d,J=7.0Hz,3H),0.90-0.80(m,6H).HRMS(ESI-TOF):Calcd for C 20 H 31 ClN3O4,[M+H] + ,412.1998,found 412.1998.

[0540] Step 2.2. Synthesis of compound I-8b: Compound I-7b (1.50 g, 3.13 mmol) and N,N-diisopropylethylamine (2.07 mL, 12.5 mmol) were dissolved in acetonitrile (30 mL), and ethylsulfonyl chloride (356 μL, 3.76 mmol) was added. The reaction was carried out at room temperature for 2 hours. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue was purified by column chromatography (ethyl acetate / dichloromethane = 0-60%) to give the title compound I-8b (1.09 g, 70% yield). 1H NMR(400MHz, DMSO-d6)δ10.20(s,1H),8.04(d,J=7.6Hz,1H),7.62(d,J=8.5Hz,2H),7.42-7.33 (m,2H),6.77(d,J=9.0Hz,1H),6.09(t,J=5.8Hz,1H),5.46(s,2H),4.72(s,2H),4.48-4.38(m, 1H),3.87-3.79(m,1H),2.97(dh,J=25.8,6.4Hz,2H),1.98-1.90(m,1H),1.75-1.65(m,1H),1. 64-1.54(m,1H),1.48-1.35(m,11H),0.84(dd,J=16.7,6.7Hz,6H).LC-MS(ESI):m / z498.3[M+H] + .

[0541] Step 3. Synthesis of compounds I-4a and I-4b

[0542] Step 3.1. Synthesis of compound I-4a. Compound I-3 (1.00 g, 1.52 mmol) and compound I-8a (691 mg, 1.68 mmol) were dissolved in dimethylformamide (20 mL), and potassium carbonate (631 mg, 4.57 mmol) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was monitored by LC-MS until complete, the compound was poured into water, extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (methanol / dichloromethane = 0-10%) to give the title compound I-4a (610 mg, yield 39%). 1H NMR (400MHz, DMSO-d6) δ10.06(s,1H),8.99(s,1H),8.48(t,J=6.1Hz,1H),8.09(d,J=7.0Hz,1H),7.88(d,J=9.3Hz,1H),7.62(d,J=8.6Hz,2 H),7.49(d,J=7.8Hz,1H),7.43(d,J=8.3Hz,2H),7.10(d,J=1.7Hz,1H),6.95-6.89(m,1H),6.74(d,J=8.8Hz,1H),5.17-5.12(m,2H),4.59-4 .31(m,5H),4.19(dd,J=16.8,5.4Hz,1H),3.88-3.80(m,1H),3.71-3.60(m,2H),3.33-3.28(m,2H),2.41(s,3H),2.32-2.22(m,1H),2.15-2 .08(m,1H),2.05-2.00(m,1H),1.98-1.87(m,2H),1.57-1.41(m,5H),1.38(s,9H),1.33-1.21(m,15H),0.96-0.80(m,15H).LC-MS(ESI):m / z 1031.5[M+H] + .

[0543] Step 3.2. Synthesis of compound I-4b:

[0544] Compound I-3 (650 mg, 0.99 mmol) and compound I-8b (543 mg, 1.09 mmol) were dissolved in dimethylformamide (10 mL), and potassium carbonate (410 mg, 2.97 mmol) was added. The mixture was reacted at room temperature for 1 hour. After the reaction was monitored by LC-MS until complete, the mixture was poured into water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (methanol / dichloromethane = 0-10%) to give the title compound I-4b (472 mg, yield 41%). LC-MS (ESI): m / z 1117.6 [M+H] + .

[0545] Step 4. Synthesis of compounds I-6a and I-6b

[0546] Step 4.1. Synthesis of Compound I-6a. Compound I-4a (600 mg, 0.58 mmol) and triphenylphosphine (458 mg, 1.75 mmol) were dissolved in tetrahydrofuran (15 mL) and water (1 mL), and reacted overnight at room temperature under nitrogen protection. After the reaction was completed by LC-MS monitoring, the mixture was concentrated under reduced pressure. The residue, compound 16 (306 mg, 0.61 mmol), and N,N-diisopropylethylamine (288 μL, 1.75 mmol) were dissolved in dichloromethane (15 mL) and methanol (2 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (243 mg, 0.64 mmol) was added, and the mixture was reacted at room temperature for 2 hours. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue was purified by column chromatography (methanol / dichloromethane = 0-12%) to give the title compound I-6a (476 mg, yield 55%, 2 steps). 1 H NMR (400MHz, DMSO-d6) δ11.93(s,1H),10.06(s,1H),8.98(s,1H),8.48(t,J=6.0H z,1H),8.41(t,J=5.7Hz,1H),8.12-8.05(m,2H),7.92(s,1H),7.87(d,J=9.3Hz,1 H),7.76(s,1H),7.66-7.58(m,3H),7.49(d,J=7.8Hz,1H),7.43(d,J=8.3Hz,2H), 7.30-7.25(m,2H),7.11-7.07(m,1H),6.92(d,J=8.3Hz,1H),6.74(d,J=8.9Hz,1H ),5.95(s,1H),5.18-5.12(m,3H),4.58-4.32(m,6H),4.24-4.15(m,1H),3.88-3. 80(m,1H),3.68-3.61(m,5H),3.18-3.15(m,2H),2.91(s,3H),2.40(s,3H),2.31- 2.22(m,1H),2.16-1.85(m,6H),1.52-1.41(m,4H),1.38(s,9H),1.31(d,J=7.0Hz ,3H),1.26-1.19(m,12H),0.92(s,9H),0.88-0.80(m,6H).HRMS(ESI-TOF):Calcd for C 76 H 97 F2N 12 O 13 S2,[M+H] + ,1487.6702,found 1487.6701.

[0547] Step 4.2. Synthesis of Compound I-6b. Compound I-4a (420 mg, 0.38 mmol) and triphenylphosphine (296 mg, 1.18 mmol) were dissolved in tetrahydrofuran (10 mL) and water (1 mL), and reacted overnight at room temperature under nitrogen protection. After the reaction was completed by LC-MS monitoring, the mixture was concentrated under reduced pressure. The residue, compound 16 (190 mg, 0.38 mmol), and N,N-diisopropylethylamine (251 μL, 1.52 mmol) were dissolved in dichloromethane (5 mL) and methanol (1 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (159 mg, 0.42 mmol) was added, and the mixture was reacted at room temperature for 2 hours. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue was purified by column chromatography (methanol / dichloromethane = 0-12%) to give the title compound I-6b (471 mg, yield 79%, 2 steps). 1 H NMR (400MHz, DMSO-d6) δ11.95(s,1H),10.11(s,1H),8.98(s,1H),8.49(d,J=6.7Hz ,1H),8.42(t,J=5.6Hz,1H),8.08(d,J=2.5Hz,1H),8.02(d,J=7.6Hz,1H),7.90(d,J =18.3Hz,2H),7.77(s,1H),7.66-7.59(m,3H),7.50(d,J=7.8Hz,1H),7.43(d,J=8. 2Hz,2H),7.30-7.26(m,2H),7.09(s,1H),6.92(d,J=7.9Hz,1H),6.78(d,J=8.9Hz,1 H),5.99(t,J=5.8Hz,1H),5.43(s,2H),5.19-5.12(m,3H),4.60-4.31(m,6H),4.23 -4.15(m,1H),3.84(t,J=7.9Hz,1H),3.64(d,J=8.5Hz,5H),3.15(s,2H),3.06-2.94 (m,2H),2.91(s,3H),2.41(s,3H),2.32-2.22(m,1H),2.15-1.87(m,6H),1.73-1.56 (m,2H),1.38(s,16H),1.23(d,J=6.0Hz,12H),0.94-0.81(m,15H).LC-MS(ESI):m / z 1573.7[M+H] + .

[0548] Step 5. Synthesis of XZ1509, XZ1512, XZ1510 and XZ1513

[0549] Step 5.1. Synthesis of compound XZ1509. Compound I-6a (40 mg, 0.03 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (1 mL) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure, and the residue was slurried with diethyl ether to obtain an intermediate. This intermediate, compound I-9 (6 mg, 0.03 mmol), and N,N-diisopropylethylamine (18 μL, 0.12 mmol) were dissolved in dimethyl sulfoxide (2 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (11 mg, 0.04 mmol) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was purified by preparative liquid chromatography to obtain the title compound XZ1509 (41 mg, 96% yield, 2 steps). 1 H NMR (400MHz, DMSO-d6) δ11.93(s,1H),9.98(s,1H),8.97(s,1H),8.48(t,J=6. 0Hz,1H),8.41(t,J=5.7Hz,1H),8.18(d,J=7.0Hz,1H),8.07(d,J=2.5Hz,1H),7 .92(s,1H),7.85(dd,J=17.2,9.0Hz,2H),7.76(s,1H),7.65-7.58(m,3H),7.4 9(d,J=7.8Hz,1H),7.43(d,J=8.2Hz,2H),7.29-7.26(m,2H),7.09(s,1H),7.00 (s,2H),6.91(d,J=8.2Hz,1H),5.14(d,J=3.5Hz,3H),4.54(d,J=9.4Hz,1H),4 .49-4.44(m,1H),4.41-4.33(m,3H),4.25-4.11(m,3H),3.64(d,J=8.6Hz,6H), 2.91(s,3H),2.40(s,3H),2.31-1.87(m,9H),1.57-1.38(m,10H),1.31(d,J=7. 1Hz,4H),1.25-1.19(m,15H),0.92(s,9H),0.87-0.81(m,6H).LC-MS(ESI):m / z 1580.7 [M+H] + .

[0550] Step 5.2. Synthesis of compound XZ1512. Compound I-6b (30 mg, 0.02 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (1 mL) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure, and the residue was slurried with diethyl ether to obtain an intermediate. This intermediate, compound I-9 (5 mg, 0.04 mmol), and N,N-diisopropylethylamine (12 μL, 0.08 mmol) were dissolved in dimethyl sulfoxide (2 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (11 mg, 0.03 mmol) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was purified by preparative liquid chromatography to obtain the title compound XZ1512 (22 mg, 70% yield, 2 steps). 1H NMR(600MHz,DMSO-d6)δ11.92(d,J=2.7Hz,1H),10.01(s,1H),8.97(s,1H),8.47(t,J=6.2Hz,1H),8.40(t,J=5.7Hz,1H),8.10-8.06(m,2H),7.92(s,1H),7.85(d,J=9.4Hz,1H),7.80(d,J=8.6Hz,1H),7.75(s,1H),7.65-7.58(m,3H),7.49(d,J=7.7Hz,1H),7.42(d,J=8.6Hz,2H),7.27(d,J=3.2Hz,2H),7.09(d,J=1.7Hz,1H),6.99(s,2H),6.91(dd,J=7.7,1.5Hz,1H),6.00-5.96(m,1H),5.41(s,2H),5.15(s,3H),4.54(d,J=9.4Hz,1H),4.46(t,J=8.0Hz,1H),4.42-4.32(m,3H),4.22-4.16(m,2H),3.63(s,5H),3.38-3.35(m,4H),3.15(s,2H),3.06-2.99(m,1H),2.97-2.92(m,1H),2.91(s,3H),2.40(s,3H),2.30-2.23(m,1H),2.21-2.15(m,1H),2.14-2.08(m,2H),2.05-2.00(m,1H),1.97(q,J=6.8Hz,1H),1.93-1.88(m,1H),1.75-1.66(m,1H),1.64-1.56(m,1H),1.54-1.40(m,10H),1.39-1.33(m,1H),1.27-1.16(m,15H),0.92(s,9H),0.84(dd,J=19.6,6.8Hz,6H).LC-MS(ESI):m / z1666.8[M+H] + .

[0551] Step 5.3. Synthesis of compound XZ1510. Compound I-6a (50 mg, 0.04 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (1 mL) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure, and the residue was slurried with diethyl ether to obtain an intermediate. This intermediate, compound I-5 (20 mg, 0.05 mmol), and N,N-diisopropylethylamine (23 μL, 0.16 mmol) were dissolved in dimethyl sulfoxide (2 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (16 mg, 0.05 mmol) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was purified by preparative liquid chromatography to obtain the title compound XZ1510 (20 mg, 31% yield, 2 steps). 1 H NMR (400MHz, DMSO-d6) δ11.94(s,1H),9.97(s,1H),8.98(s,1H),8.48(s,1H),8.41(t,J=5.6Hz,1H),8.20(d,J=7.0Hz,1H),8.08(d,J=2.5Hz,1H),7 .94-7.85(m,3H),7.76(s,1H),7.65-7.59(m,3H),7.49(d,J=7.8Hz,1H), 7.43(d,J=8.2Hz,2H),7.30-7.26(m,2H),7.09(s,1H),7.02(s,1H),6.94- 6.89(m,1H),5.17-5.12(m,3H),4.55(d,J=9.4Hz,1H),4.46(t,J=8.1Hz, 1H),4.43-4.32(m,4H),4.24-4.14(m,3H),3.68-3.42(m,35H),2.91(s,3H) ),2.40(s,3H),2.37-2.21(m,2H),2.14-1.89(m,5H),1.45(s,4H),1.34- 1.20(m,16H),0.92(s,9H),0.86(dd,J=16.5,6.7Hz,6H).LC-MS(ESI):m / z 1802.9[M+H] + .

[0552] Step 5.4. Synthesis of compound XZ1513. Compound I-6b (30 mg, 0.02 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (1 mL) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure, and the residue was slurried with diethyl ether to obtain an intermediate. This intermediate, compound I-5 (15 mg, 0.04 mmol), and N,N-diisopropylethylamine (17 μL, 0.12 mmol) were dissolved in dimethyl sulfoxide (2 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (13 mg, 0.04 mmol) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was purified by preparative liquid chromatography to obtain the title compound XZ1513 (10 mg, yield 29%, 2 steps). LC-MS(ESI): m / z 1888.8 [M+H] + .

[0553] Step 6. Synthesis of compound GNE-987-OH. Compound I-3 (20 mg, 0.03 mmol) and triphenylphosphine (10 mg, 0.04 mmol) were dissolved in tetrahydrofuran (2 mL) and water (200 μL), and reacted overnight at room temperature under nitrogen protection. After the reaction was completed by LC-MS monitoring, the solution was concentrated under reduced pressure. The residue, compound 16 (15 mg, 0.03 mmol), and N,N-diisopropylethylamine (15 μL, 0.09 mmol) were dissolved in dichloromethane (2 mL) and methanol (200 μL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (19 mg, 0.05 mmol) was added, and the reaction was carried out at room temperature for 2 hours. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue was purified by column chromatography (methanol / dichloromethane = 0-8%) to give the title compound (21 mg, yield 63%, 2 steps). 1H NMR (400MHz, DMSO-d6) δ11.93(d,J=2.7Hz,1H),9.79(s,1H),8.96(s,1H),8.49-8.36(m,2H),8.07(d,J=2.5Hz,1H),7.92(s,1H),7.8 4(d,J=9.3Hz,1H),7.76(s,1H),7.66-7.57(m,1H),7.37(d,J=7.8Hz,1H),7.30-7.25(m,2H),6.91(d,J=1.8Hz,1H),6.80(dd,J=7.8, 1.8Hz,1H),4.58-4.50(m,1H),4.48-4.42(m,1H),4.38-4.24(m,2H),4.17-4.09(m,1H),3.67-3.61(m,5H),3.20-3.11(m,2H),2.91( s,3H),2.45(s,3H),2.31-2.21(m,1H),2.16-1.87(m,4H),1.52-1.39(m,4H),1.33-1.17(m,16H),0.92(s,9H).HRMS(ESI-TOF):Calcd for C56H68F2N9O9S2,[M+H]+,1112.4544,found 1112.4542.

[0554] Scheme 2. Synthesis of XZ1522, XZ1522A, XZ1522B, XZ1522C, XZ1532A and XZ1532B

[0555]

[0556] Step 1. Synthesis of Compound II-2. Compound II-1 (250 mg, 0.46 mmol) was dissolved in tetrahydrofuran (10 mL), and thionyl chloride (450 μL, 6.20 mmol) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue was slurried with diethyl ether to give the title compound II-2 (217 mg, yield 84%), which was directly added to the next step. LC-MS (ESI): m / z 566.3 [M+H] + .

[0557] Step 2. Synthesis of Compound II-3. Compound I-3 (91 mg, 0.14 mmol) and Compound II-2 (78 mg, 0.14 mmol) were dissolved in dimethylformamide (20 mL), and potassium carbonate (135 mg, 0.42 mmol) was added. The reaction was carried out at room temperature for 30 minutes. After the reaction was monitored by LC-MS until complete, the compound was poured into water, extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was directly used in the next step. LC-MS (ESI): m / z 1185.6 [M+H] + .

[0558] Step 3. Synthesis of Compound II-4. Compound II-3 (0.138 mmol) and triphenylphosphine (73 mg, 0.28 mmol) were dissolved in tetrahydrofuran (8 mL) and water (50 μL), and reacted overnight at room temperature under nitrogen protection. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure. The residue, compound 16 (34 mg, 0.068 mmol), and N,N-diisopropylethylamine (44 μL, 0.27 mmol) were dissolved in dichloromethane (10 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (27 mg, 0.071 mmol) were added. The mixture was reacted at room temperature for 30 min. After the reaction was completed by LC-MS, the mixture was purified by preparative liquid chromatography to give the title compound II-4 (61 mg, 55% yield). 1H NMR(600MHz,DMSO-d6)δ11.93(s,1H),10.13(s,1H),8.99(s,1H),8.48-8.45(m,1H),8.43-8.38(m,1 H),8.08(d,J=2.5Hz,1H),8.00(d,J=7.5Hz,1H),7.92(s,1H),7.85(d,J=9.4Hz,1H),7.76(s,1H),7.6 7(d,J=2.2Hz,1H),7.65-7.59(m,2H),7.51-7.44(m,2H),7.29-7.26(m,2H),7.12(s,1H),6.94(d,J=8 .0Hz,1H),6.76(d,J=8.8Hz,1H),6.00-5.94(m,1H),5.35-5.31(m,1H),5.20(s,2H),4.67(s,2H),4.5 7-4.53(m,1H),4.48-4.42(m,2H),4.38-4.32(m,2H),4.24(d,J=2.4Hz,2H),4.21-4.18(m,1H),3.85- 3.82(m,1H),3.65-3.63(m,7H),3.43(t,J=2.4Hz,1H),3.18-3.12(m,2H),3.03-3.00(m,1H),2.97-2. 93(m,1H),2.91(s,3H),2.43(s,3H),2.29-2.24(m,1H),2.14-2.10(m,1H),2.03-1.91(m,4H),1.73-1 .68(m,1H),1.64-1.57(m,1H),1.45-1.22(m,30H),0.94-0.81(m,15H).LC-MS(ESI):m / z1641.7[M+H] + .

[0559] Step 4. General synthetic route for XZ1522 and XZ1522A-C

[0560] Compound II-4 (1.0 equivalent), the corresponding azide compound (1.5 equivalent), copper sulfate pentahydrate (0.2 equivalent), and sodium ascorbate (0.4 equivalent) were dissolved in 1,4-dioxane (3 mL), tert-butanol (2 mL), and water (1 mL), and reacted at 50 °C for 1 hour under nitrogen protection. After the reaction was completed by LC-MS, the solution was concentrated under reduced pressure. The residue and trifluoroacetic acid (20 equivalent) were dissolved in dichloromethane (5 mL), and reacted at room temperature for 30 minutes. After the reaction was completed by LC-MS, the solution was concentrated under reduced pressure. The residue, compound I-9 (1.0 equivalent), and N,N-diisopropylethylamine (5.0 equivalent) were dissolved in dimethyl sulfoxide (3 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.05 equivalent) was added, and the solution was reacted at room temperature for 20 minutes. After the reaction was monitored to be complete by LC-MS, the title compound was obtained by preparative liquid chromatography.

[0561] Step 4.1. Synthesis of compound XZ1522. Using azide compound II-6 (6.4 mg) as a starting material, the title compound XZ1522 was obtained via a general synthetic route. 1H NMR(600MHz,DMSO-d6)δ11.92(s,1H),10.05(s,1H),8.98(s,1H),8.49-8.45(m,1H),8.42-8.38(m,1H) ,8.10-8.05(m,3H),7.92(s,1H),7.85(d,J=9.3Hz,1H),7.80(d,J=8.7Hz,1H),7.76(s,1H),7.67-7.58 (m,3H),7.50(d,J=7.8Hz,1H),7.45(d,J=8.4Hz,1H),7.30-7.27(m,2H),7.09(d,J=1.6Hz,1H),7.00(s ,2H),6.93(d,J=7.9Hz,1H),5.99-5.95(m,1H),5.43-5.38(m,1H),5.35-5.31(m,1H),5.18(s,2H),4.66 (s,2H),4.61(s,2H),4.55(d,J=9.4Hz,1H),4.48-4.44(m,3H),4.41-4.32(m,3H),4.22-4.14(m,2H),3 .78(t,J=5.3Hz,2H),3.68-3.60(m,5H),3.51-3.44(m,88H),3.24(s,3H),3.19-3.11(m,2H),3.04-2.93 (m,2H),2.91(s,3H),2.40(s,3H),2.31-2.24(m,1H),2.22-2.16(m,1H),2.14-2.07(m,2H),2.06-1.88 (m,4H),1.74-1.67(m,1H),1.63-1.56(m,1H),1.50-1.19(m,27H),0.93-0.81(m,15H).LC-MS(ESI):m / z 1381.2[(M+2H) / 2] + .

[0562] Step 4.2. Synthesis of compound XZ1522A. Using azide compound II-7 (8.6 mg) as a starting material, the title compound XZ1522A was obtained via a general synthetic route. 1H NMR(400MHz,DMSO-d6)δ11.93(s,1H),10.05(s,1H),8.98(s,1H),8.50-8.36(m, 2H),8.11-8.05(m,3H),7.92(s,1H),7.89-7.77(m,2H),7.76(s,1H),7.68-7.58 (m,3H),7.52-7.42(m,2H),7.31-7.25(m,2H),7.09(s,1H),7.00(s,2H),6.92(d ,J=8.0Hz,1H),6.01-5.93(m,1H),5.35-5.31(m,1H),5.18(s,2H),4.66(s,2H),4 .61(s,2H),4.55(d,J=9.2Hz,1H),4.50-4.43(m,3H),4.42-4.30(m,3H),4.23-4 .14(m,2H),3.78(t,J=5.3Hz,2H),3.68-3.62(m,5H),3.51-3.43(m,16H),3.20( s,3H),3.17-3.12(m,2H),3.05-2.91(m,2H),2.91(s,3H),2.40(s,3H),2.31-1. 68(m,10H),1.46-1.23(m,27H),0.92(s,9H),0.86-0.82(m,6H).LC-MS(ESI):m / z 984.6[(M+2H) / 2] + .

[0563] Step 4.3. Synthesis of compound XZ1522B. Using azide compound II-8 (10.4 mg) as a starting material, the title compound XZ1522B was obtained via a general synthetic route. 1H NMR(400MHz,DMSO-d6)δ11.93(s,1H),10.05(s,1H),8.98(s,1H),8.51-8.36(m, 2H),8.11-8.05(m,3H),7.92(s,1H),7.88-7.78(m,2H),7.76(s,1H),7.68-7.56 (m,3H),7.52-7.42(m,2H),7.31-7.25(m,2H),7.09(s,1H),7.00(s,2H),6.92(d ,J=7.9Hz,1H),6.02-5.93(m,1H),5.34-5.32(m,1H),5.18(s,2H),4.66(s,2H), 4.61(s,2H),4.55(d,J=9.4Hz,1H),4.49-4.43(m,3H),4.42-4.30(m,3H),4.24- 4.12(m,2H),3.78(t,J=5.4Hz,2H),3.69-3.60(m,5H),3.50-3.41(m,36H),3.23 (s,3H),3.19-3.12(m,2H),3.06-2.94(m,2H),2.91(s,3H),2.40(s,3H),2.27-1 .99(m,8H),1.74-1.25(m,29H),0.92(s,9H),0.86-0.80(m,6H).LC-MS(ESI):m / z 1095.0[(M+2H) / 2] + .

[0564] Step 4.4. Synthesis of compound XZ1522C. Using azide compound II-9 (4.5 mg) as a starting material, the title compound XZ1522C was obtained via a general synthetic route. 1H NMR(400MHz,DMSO-d6)δ11.92(s,1H),10.05(s,1H),8.97(s,1H),8.50-8.37( m,2H),8.11-8.06(m,3H),7.92(s,1H),7.88-7.78(m,2H),7.76(s,1H),7.68-7 .57(m,3H),7.53-7.43(m,2H),7.30-7.25(m,2H),7.09(s,1H),7.00(s,2H),6. 92(d,J=7.9Hz,1H),6.01-5.93(m,1H),5.44-5.30(m,2H),5.18(s,2H),4.66(s ,2H),4.61(s,2H),4.55(d,J=9.3Hz,1H),4.49-4.43(m,3H),4.41-4.31(m,3H) ,4.22-4.13(m,2H),3.78(t,J=5.3Hz,2H),3.67-3.61(m,5H),3.54-3.42(m,96 H),3.24(s,3H),3.19-3.12(m,2H),3.05-2.93(m,2H),2.91(s,3H),2.40(s,3H ),2.22-1.94(m,8H),1.67-1.23(m,29H),0.93-0.81(m,15H).LC-MS(ESI):m / z 1425.2[(M+2H) / 2] + .

[0565] Step 5. General Synthetic Route for XZ1532A and XZ1532B

[0566] Compound II-4 (1.0 equivalent) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (1 mL) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed by LC-MS, the solution was concentrated under reduced pressure. The residue, compound II-5 (1.0 equivalent), and N,N-diisopropylethylamine (5.0 equivalent) were dissolved in dichloromethane (5 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.05 equivalent) was added. The reaction was carried out at room temperature for 20 minutes. After the reaction was completed by LC-MS, the solution was concentrated under reduced pressure. The residue, the corresponding azide compound (1.5 equivalent), copper sulfate pentahydrate (0.2 equivalent), and sodium ascorbate (0.4 equivalent) were dissolved in 1,4-dioxane (3 mL), tert-butanol (2 mL), and water (1 mL). The reaction was carried out at 50 °C for 2 hours under nitrogen protection. After the reaction was monitored to be complete by LC-MS, the title compound was obtained by preparative liquid chromatography.

[0567] Step 5.1. Synthesis of compound XZ1532A. Using azide compound II-9 (3.2 mg) as a starting material, the title compound XZ1532A was obtained via a general synthetic route. 1 H NMR(600MHz,DMSO-d6)δ11.92(s,1H),10.04(s,1H),8.97(s,1H),8.50-8.37(m,2H),8.14 -8.04(m,3H),7.92(s,1H),7.84(d,J=8.9Hz,2H),7.76(s,1H),7.69-7.57(m,3H),7.53-7 .43(m,2H),7.28(s,2H),7.09(s,1H),7.02-6.99(m,2H),6.93(d,J=7.8Hz,1H),5.99-5.9 4(m,1H),5.39(s,2H),5.18(s,2H),5.11(s,1H),4.66(s,2H),4.61(s,2H),4.55(d,J=9.2H z,1H),4.49-4.43(m,3H),4.41-4.32(m,3H),4.25-4.20(m,1H),4.19-4.15(m,1H),3.80- 3.75(m,2H),3.67-3.62(m,5H),3.56-3.43(m,100H),3.24(s,3H),3.19-3.10(m,2H),3.05 -2.99(m,1H),2.99-2.94(m,1H),2.91(s,3H),2.46-2.38(m,4H),2.35-2.23(m,2H),2.17 -2.06(m,1H),2.05-1.86(m,4H),1.70-1.23(m,23H),0.94-0.81(m,15H).LC-MS(ESI):m / z 1426.2[(M+2H) / 2] + .

[0568] Step 5.2. Synthesis of compound XZ1532B. Using azide compound II-10 (1.2 mg) as a starting material, the title compound XZ1532B was obtained via a general synthetic route. 1H NMR(600MHz,DMSO-d6)δ12.15(s,1H),11.92(s,1H),10.04(s,1H),8.98(s,1H),8.49-8.4 4(m,1H),8.42-8.37(m,1H),8.12-8.06(m,3H),7.92(s,1H),7.87-7.81(m,2H),7.76(s,1H ),7.69-7.59(m,3H),7.52-7.43(m,2H),7.30-7.26(m,2H),7.09(s,1H),7.01(s,2H),6.93 (d,J=7.7Hz,1H),5.99-5.93(m,1H),5.39(s,2H),5.18(s,2H),5.11(d,J=3.5Hz,1H),4.66 (s,2H),4.61(s,2H),4.55(d,J=9.4Hz,1H),4.48-4.45(m,3H),4.41-4.32(m,3H),4.24-4 .14(m,2H),3.78(t,J=5.4Hz,2H),3.68-3.63(m,5H),3.61-3.46(m,100H),3.18-3.11(m,2 H),3.05-2.93(m,2H),2.91(s,3H),2.46-2.38(m,7H),2.35-2.23(m,2H),2.14-2.08(m,1H ),2.05-1.87(m,5H),1.69-1.22(m,23H),0.92(s,9H),0.86-0.80(m,6H).LC-MS(ESI):m / z 1455.3[(M+2H) / 2] + .

[0569] Step 6. General synthetic route for compounds 51 and 51'

[0570] Compound 50 (1.0 equivalent) and propynyl alcohol / butynyl alcohol (5.0 equivalent) were dissolved in acetonitrile (15 mL), and cesium carbonate (3.0 equivalent) was added. The reaction was carried out overnight at room temperature. After the reaction was monitored by LC-MS until complete, the mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (ethyl acetate / petroleum ether = 0-20%) to obtain the corresponding title compound, which was directly used in the next step.

[0571] Step 7. General synthetic route for compounds II-1 and II-1'

[0572] Compound 51 / 51' (1.0 equivalent) was dissolved in methanol (10 mL) and water (2 mL), and iron powder (5.0 equivalent) and ammonium chloride (5.0 equivalent) were added. The mixture was refluxed overnight at 65 °C. After the reaction was complete as monitored by TLC, the mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in tetrahydrofuran (20 mL), and lithium aluminum hydride (2.5 equivalent) was added under ice bath conditions. The mixture was then reacted at room temperature for 4 hours. After the reaction was complete as monitored by TLC, water (1 mL) and saturated sodium hydroxide solution (1 mL) were added to quench the reaction. The mixture was dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The residue and compound 52 (1.0 equivalent) were dissolved in dichloromethane (10 mL), and EEDQ (2.0 equivalent) was added. The mixture was reacted overnight at room temperature. After the reaction was complete as monitored by TLC, the mixture was concentrated under reduced pressure. The residue was purified by column chromatography to obtain the corresponding title compound.

[0573] Step 7.1. Synthesis of compound II-1. The title compound II-1 was obtained via a general synthetic route. 1 H NMR (400MHz, DMSO-d6) δ10.01 (s, 1H), 7.97 (d, J = 7.7Hz, 1H), 7.58-7.50 (m, 2H), 7.35-7.28 (m, 1H ),6.76(d,J=8.9Hz,1H),5.96(t,J=5.9Hz,1H),5.39(s,2H),5.05-4.99(m,1H),4.55(s,2H),4.5 2-4.37(m,3H),4.21(d,J=2.4Hz,2H),3.87-3.79(m,1H),3.50(t,J=2.4Hz,1H),3.07-2.88(m,2H ),2.02-1.90(m,1H),1.75-1.51(m,2H),1.45-1.36(m,11H),0.89-0.81(m,6H).LC-MS(ESI):m / z 548.3[M+H] + .

[0574] Step 7.2. Synthesis of compound II-1'. The title compound II-1' was obtained via a general synthetic route. 1H NMR(600MHz,DMSO-d6)δ9.99(s,1H),7.97(d,J=7.7Hz,1H),7.59-7.50(m,2H),7.32(d,J=8.8Hz,1H),6.76( d,J=8.9Hz,1H),5.96(s,1H),5.39(s,2H),5.02-4.95(m,1H),4.53-4.47(m,4H),4.43(d,J=7.6Hz,1H),3.8 7-3.81(m,1H),3.57-3.52(m,2H),3.06-2.91(m,2H),2.82-2.79(m,1H),2.48-2.43(m,2H),1.99-1.92(m,1 H),1.68(s,1H),1.63-1.56(m,1H),1.38(d,J=12.8Hz,11H),0.85(dd,J=24.1,6.7Hz,6H).LC-MS(ESI):m / z 562.3[M+H] + .

[0575] Scheme 3. Synthesis of XZ1525B, XZ1528A, XZ1528B and XZ1534

[0576]

[0577] Step 1. Synthesis of compound XZ1525B. Compound XZ1729-C11 (10 mg, 0.009 mmol), compound III-1 (8.2 mg, 0.011 mmol), N,N-diisopropylethylamine (30 μL, 0.182 mmol), and 1-hydroxybenzotriazole (1.5 mg, 0.011 mmol) were dissolved in dimethylformamide (2 mL) and reacted at room temperature for 2 hours. After the reaction was completed as monitored by LC-MS, the title compound XZ1525B (7.3 mg, 57%) was purified by preparative liquid chromatography. 1H NMR (600MHz, DMSO-d6) δ11.87(s,1H),9.99(s,1H),8.98(s,1H),8.56(t,J=6.2Hz,1H),8.13-8.05(m,2H),7.87-7.77(m,3H),7 .66(s,1H),7.63-7.51(m,2H),7.44-7.37(m,4H),7.26-7.13(m,3H),6.99(s,2H),5.98(s,1H),5.35-5.31(m,1H),4.97(s,1H), 4.64-4.52(m,2H),4.47-4.32(m,4H),4.25-4.17(m,2H),3.69-3.60(m,5H),3.07-2.92(m,7H),2.57-2.54(m,6H),2.44(s,3H) ,2.29-2.16(m,2H),2.15-2.06(m,2H),2.03-1.88(m,6H),1.57-1.23(m,25H),0.93(s,9H),0.85-0.80(m,6H).LC-MS(ESI):m / z 841.0[(M+2H) / 2] + .

[0578] Step 2. Synthesis of Compound III-3. Using compounds XZ1707-C11 and III-2 as starting materials, the title compound was obtained via the same synthetic route as XZ1525B. LC-MS (ESI): m / z 1600.6 [M+H] + .

[0579] Step 3. Synthesis of Compound III-5. Using compounds XZ1707-C11 and III-4 as starting materials, the title compound was obtained via the same synthetic route as XZ1525B. LC-MS (ESI): m / z 1668.7 [M+H] + .

[0580] Step 4. Synthesis of Compound III-6. Using compounds XZ1707-C10 and III-4 as starting materials, the title compound was obtained via the same synthetic route as XZ1525B. LC-MS (ESI): m / z 1654.7 [M+H] + .

[0581] Step 5. Synthesis of compound XZ1528A. Compound III-3 (40 mg, 0.02 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (1 mL) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure, and the residue was slurried with diethyl ether to obtain an intermediate. This intermediate, compound I-5 (26 mg, 0.05 mmol), and N,N-diisopropylethylamine (16 μL, 0.10 mmol) were dissolved in dimethyl sulfoxide (2 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (14 mg, 0.04 mmol) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was purified by preparative liquid chromatography to obtain the title compound XZ1528A (21 mg, 44% yield, 2 steps). 1 H NMR (600MHz, DMSO-d6) δ11.83(s,1H),9.98(d,J=51.6Hz,1H),8.99(s,1H),8.38(d,J=7.6Hz,1H),8 .12(d,J=17.8Hz,1H),8.01(t,J=5.8Hz,1H),7.87(s,1H),7.83-7.76(m,2H),7.68(s,1H),7.62(d, J=8.1Hz,1H),7.50(d,J=8.3Hz,1H),7.44(d,J=8.0Hz,2H),7.39(d,J=8.1Hz,2H),7.32(dd,J=20.3 ,7.6Hz,1H),7.25-7.17(m,2H),7.07(d,J=7.0Hz,1H),7.00(s,3H),6.59(d,J=50.9Hz,1H),6.21(s, 1H),5.99(s,1H),5.10-4.89(m,4H),4.63-4.19(m,11H),3.64-3.57(m,12H),3.51-3.49(m,22H),3 .15(q,J=5.8Hz,3H),3.10-2.92(m,7H),2.46(s,3H),2.33(t,J=7.3Hz,2H),2.28-2.22(m,1H),2.13 -2.06(m,1H),2.05-1.94(m,2H),1.84-1.77(m,1H),1.68(d,J=24.0Hz,1H),1.60(s,1H),1.53-1.2 9(m,10H),1.26-1.05(m,14H),0.93(s,10H),0.87-0.82(m,6H).LC-MS(ESI):m / z1016.2[(M+2H) / 2] + .

[0582] Step 6. Synthesis of compound XZ1528B. Compound III-5 (30 mg, 0.02 mmol), compound II-9 (42 mg, 0.04 mmol), copper sulfate pentahydrate (1 mg, 0.01 mmol), and sodium ascorbate (1 mg, 0.01 mmol) were dissolved in 1,4-dioxane (3 mL), tert-butanol (2 mL), and water (1 mL). The mixture was reacted at 50 °C for 1 hour under nitrogen protection. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue and trifluoroacetic acid (1 mL) were dissolved in dichloromethane (5 mL), and the mixture was reacted at room temperature for 30 minutes. After the reaction was completed under LC-MS monitoring, the solution was concentrated under reduced pressure. The residue, compound II-5 (37 mg, 0.02 mmol), and N,N-diisopropylethylamine (10 μL, 0.50 mmol) were dissolved in dimethyl sulfoxide (3 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (10 mg, 0.03 mmol) was added, and the mixture was reacted at room temperature for 20 minutes. After the reaction was completed under LC-MS monitoring, the solution was purified by preparative liquid chromatography to give the title compound XZ1528B (18 mg, 32% yield, 3 steps). 1 H NMR(600MHz,DMSO-d6)δ11.84(s,1H),10.00(d,J=47.6Hz,1H),8.99(s,1H),8.37 (d,J=7.8Hz,1H),8.06(d,J=31.3Hz,2H),7.88-7.75(m,2H),7.68(s,1H),7.62(s ,1H),7.54-7.42(m,2H),7.38(d,J=8.0Hz,2H),7.34-7.21(m,1H),7.17(s,1H),7 .00(s,3H),6.21(s,1H),5.96(s,1H),5.41(s,1H),5.04(d,J=17.2Hz,2H),4.96-4 .89(m,1H),4.66-4.16(m,16H),3.81(t,J=5.2Hz,2H),3.60(s,7H),3.52-3.44(m ,105H),3.24(s,3H),3.09(s,1H),3.04-2.87(m,5H),2.46(s,3H),2.33(s,1H),2 .28-2.21(m,1H),2.15-1.92(m,2H),1.83-1.78(m,1H),1.72-1.57(m,2H),1.52- 1.30(m,9H),1.25-1.04(m,13H),0.93(s,9H),0.86-0.79(m,6H).LC-MS(ESI):m / z 1439.3[(M+2H) / 2]+ .

[0583] Step 7. Synthesis of compound XZ1534. Compound III-6 (20 mg, 0.01 mmol), compound II-10 (28 mg, 0.02 mmol), copper sulfate pentahydrate (1 mg, 0.01 mmol), and sodium ascorbate (2 mg, 0.01 mmol) were dissolved in 1,4-dioxane (3 mL), tert-butanol (2 mL), and water (1 mL). The mixture was reacted at 50 °C for 1 hour under nitrogen protection. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure. The residue and trifluoroacetic acid (1 mL) were dissolved in dichloromethane (5 mL), and the mixture was reacted at room temperature for 30 minutes. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure. The residue, compound II-11 (4 mg, 0.01 mmol), and N,N-diisopropylethylamine (10 μL, 0.50 mmol) were dissolved in dimethyl sulfoxide (3 mL), and the mixture was reacted at room temperature for 20 minutes. After the reaction was monitored to be complete by LC-MS, the title compound XZ1534 (9 mg, yield 24%, 3 steps) was purified by preparative liquid chromatography. 1 H NMR(600MHz,DMSO-d6)δ11.83(d,J=10.8Hz,1H),10.01(d,J=53.6Hz,1H),8. 99(s,1H),8.38(s,1H),8.09(t,J=26.1Hz,2H),7.90-7.75(m,3H),7.65(d,J =31.7Hz,2H),7.50(d,J=17.6Hz,1H),7.44(d,J=8.0Hz,2H),7.39(d,J=8.0H z,2H),7.32(d,J=25.2Hz,1H),7.24(s,1H),7.18(s,2H),7.00(s,3H),6.57( d,J=68.8Hz,1H),6.21(s,1H),5.98(s,1H),5.76(s,1H),5.13-4.89(m,4H), 4.65-4.17(m,16H),3.81(t,J=5.3Hz,3H),3.64-3.53(m,54H),3.50-3.45(m ,42H),3.21-2.90(m,9H),2.48-2.42(m,6H),2.36-2.21(m,2H),2.13-1.91( m,4H),1.84-1.01(m,29H),0.93(s,9H),0.87-0.78(m,6H).LC-MS(ESI):m / z 1461.7[(M+2H) / 2] + .

[0584] Step 8. General synthetic route for compounds III-4 and III-4'

[0585] Compound II-1 / II-1' (1.0 equivalent) was dissolved in dimethylformamide (5 mL), and diisopropylethylamine (4.0 equivalent) and di(p-nitrobenzene) carbonate (2.0 equivalent) were added. The reaction was carried out at room temperature for 3 hours. After the reaction was completed by TLC monitoring, the mixture was extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was slurried with diethyl ether to give the title compound.

[0586] Step 8.1. Synthesis of compound III-4. The title compound III-4 was obtained via a general synthetic route. 1 H NMR (400MHz, DMSO-d6) δ10.18(s,1H),8.32(d,J=9.0Hz,2H),8.02(d,J=7.5Hz,1H),7.71-7.61(m,2H),7.57(d,J=9 .0Hz,2H),7.42(d,J=8.1Hz,1H),6.75(d,J=8.9Hz,1H),5.97(t,J=5.9Hz,1H),5.42-5.37(m,2H),5.32(s,2H),4.6 2(s,2H),4.46-4.40(m,1H),4.23(d,J=2.4Hz,2H),3.84(t,J=7.7Hz,1H),3.55-3.49(m,1H),3.08-2.90(m,2H),2. 02-1.91(m,1H),1.75-1.53(m,2H),1.42-1.34(m,11H),0.85(dd,J=16.7,6.7Hz,6H).LC-MS(ESI):m / z713.4[(M+H] + .

[0587] Step 8.2. Synthesis of compound III-4'. The title compound III-4' was obtained via a general synthetic route. 1H NMR(600MHz,DMSO-d6)δ10.16(s,1H),8.35-8.29(m,2H),8.01(d,J=7.6Hz,1H),7.67-7.62(m,2H),7.60-7.56(m,2H) ,7.42(d,J=8.2Hz,1H),6.75(d,J=8.9Hz,1H),6.00-5.94(m,1H),5.40(s,2H),5.34(s,2H),4.60(s,2H),4.48-4.40( m,1H),3.87-3.82(m,1H),3.59-3.54(m,2H),3.05-2.92(m,2H),2.82-2.78(m,1H),2.48-2.45(m,2H),2.00-1.93(m, 1H),1.75-1.67(m,1H),1.65-1.57(m,1H),1.39(s,11H),0.85(dd,J=24.8,6.8Hz,6H).LC-MS(ESI):m / z727.3[(M+H] + .

[0588] Scheme 4. Synthesis of XZ1526A, XZ1539, XZ1541, XZ1526B, XZ1526C, XZ1533, XZ1537, XZ1540, XZ1542, XZ1543 and XZ1543B

[0589]

[0590] Step 1. Synthesis of compound XZ1525A. Compound XZ1717-C2 (10 mg, 0.013 mmol), compound III-1 (11.2 mg, 0.015 mmol), N,N-diisopropylethylamine (30 μL, 0.182 mmol), and 1-hydroxybenzotriazole (2.1 mg, 0.016 mmol) were dissolved in dimethylformamide (2 mL) and reacted at room temperature for 2 hours. After the reaction was completed as monitored by LC-MS, the title compound XZ1525A (9.5 mg, 54%) was purified by preparative liquid chromatography. 1H NMR(600MHz,DMSO-d6)δ11.88(s,1H),10.78(s,1H),9.99-9.95(m,1H),8.10-8.05(m,2H),7.84-7.78(m,2H),7.66-7.63(m,1H),7.57-7 .54(m,2H),7.25(d,J=2.6Hz,1H),7.22-7.17(m,3H),7.10-7.05(m,3H),6.99(s,2H),6.92-6.85(m,2H),6.68-6.63(m,1H),6.00-5.95(m ,1H),5.33(t,J=4.9Hz,1H),4.98(s,1H),4.42-4.36(m,1H),4.22-4.17(m,1H),4.04-3.96(m,1H),3.75(dd,J=11.3,4.9Hz,1H),3.61(s, 3H),3.58-3.53(m,2H),3.49-3.41(m,6H),3.26-2.92(m,7H),2.19-1.98(m,8H),1.75-1.25(m,15H),0.85-0.81(m,6H).LC-MS(ESI):m / z 1397.7[M+H] + .

[0591] Step 2. Synthesis of compound IV-1. Using compounds XZ1717-C2 and III-2 as starting materials, the title compound III-3 was obtained via the same synthetic route as XZ1525A. LC-MS (ESI): m / z 1304.5 [M+H] + .

[0592] Step 3. General synthetic route for compounds IV-2 and IV-2'.

[0593] Step 3.1. Synthesis of Compound IV-2. Using compounds XZ1717-C2 and III-4 as starting materials, the title compound IV-2 was obtained via the same synthetic route as XZ1525A. LC-MS (ESI): m / z 1372.5 [M+H] + .

[0594] Step 3.2. Synthesis of compound IV-2'. Using compounds XZ1717-C2 and III-4' as starting materials, the title compound IV-2' was obtained via the same synthetic route as XZ1525A. LC-MS (ESI): m / z 1386.5 [M+H] + .

[0595] Step 4. Synthesis of compound XZ1526A. Compound IV-1 (20 mg, 0.02 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (1 mL) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure, and the residue was slurried with diethyl ether to obtain an intermediate. This intermediate, compound I-5 (13 mg, 0.02 mmol), and N,N-diisopropylethylamine (10 μL, 0.06 mmol) were dissolved in dimethyl sulfoxide (2 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (12 mg, 0.03 mmol) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed by LC-MS, the title compound XZ1526A (9 mg, 33% yield, 2 steps) was purified by preparative liquid chromatography. 1 H NMR(400MHz,DMSO-d6)δ11.88(d,J=2.8Hz,1H),10.78(s,1H),9.97(s,1H), 8.12(d,J=7.4Hz,1H),8.06(d,J=2.5Hz,1H),8.01(t,J=5.6Hz,1H),7.87(d ,J=8.6Hz,1H),7.81(d,J=7.0Hz,1H),7.67-7.51(m,4H),7.25(d,J=2.7Hz, 1H),7.20(d,J=8.3Hz,2H),7.08(d,J=8.6Hz,3H),7.00(s,2H),6.89(t,J=9 .0Hz,2H),5.99(s,1H),4.98(s,2H),4.48(d,J=66.3Hz,5H),4.24(s,1H),3 .60(d,J=7.0Hz,10H),3.51-3.48(m,32H),3.18-2.98(m,14H),2.42-2.30( m,5H),2.21-2.08(m,1H),2.06-1.91(m,2H),1.77-1.54(m,2H),1.41(d,J= 25.4Hz,2H),0.86(d,J=6.8Hz,3H),0.83(d,J=6.8Hz,3H).LC-MS(ESI):m / z 1734.7[M+H] + .

[0596] Step 5. Synthesis of compounds XZ1526B and XZ1526C.

[0597] Step 5.1. Synthesis of compound XZ1526B. Compound IV-2 (20 mg, 0.015 mmol), compound II-9 (24 mg, 0.021 mmol), copper sulfate pentahydrate (0.73 mg, 0.003 mmol), and sodium ascorbate (1.16 mg, 0.006 mmol) were dissolved in 1,4-dioxane (3 mL), tert-butanol (2 mL), and water (1 mL). The mixture was reacted at 50 °C for 5 hours under nitrogen protection. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue and trifluoroacetic acid (1 mL) were dissolved in dichloromethane (5 mL), and the mixture was reacted at room temperature for 1 hour. After the reaction was completed as monitored by LC-MS, the solution was concentrated under reduced pressure. The residue, compound I-9 (2 mg, 0.009 mmol), and N,N-diisopropylethylamine (30 μL, 0.182 mmol) were dissolved in dimethyl sulfoxide (3 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (3.15 mg, 0.008 mmol) were added. The reaction was carried out at room temperature for 15 minutes. After the reaction was completed as monitored by LC-MS, the solution was purified by preparative liquid chromatography to give the title compound XZ1526B (13 mg, 28% yield, 3 steps). 1 H NMR(600MHz,DMSO-d6)δ11.87(s,1H),10.78(s,1H),10.05-9.97(m,1H),8.11-8.01(m,3H),7.85-7.77(m,2H),7.66-7.51(m,4H),7 .28-7.15(m,2H),7.12-7.05(m,3H),6.99(s,2H),6.93-6.85(m,2H),6.01-5.91(m,1H),5.46-5.31(m,2H),5.02(s,2H),4.58-4.45 (m,7H),4.40-4.34(m,1H),4.24-4.17(m,1H),4.06-3.92(m,1H),3.84-3.78(m,2H),3.74(dd,J=11.1,5.0Hz,1H),3.62-3.37(m,11 0H),3.24(s,3H),3.12-2.90(m,8H),2.23-2.07(m,3H),2.04-1.92(m,3H),1.74-1.17(m,11H),0.86-0.80(m,6H).LC-MS(ESI):m / z 1290.6[(M+2H) / 2] + .

[0598] Step 5.2. Synthesis of compound XZ1526C. Compound IV-2 (20 mg, 0.015 mmol), compound II-9 (24 mg, 0.021 mmol), copper sulfate pentahydrate (0.73 mg, 0.003 mmol), and sodium ascorbate (1.16 mg, 0.006 mmol) were dissolved in 1,4-dioxane (3 mL), tert-butanol (2 mL), and water (1 mL). The reaction was carried out at 50 °C for 5 hours under nitrogen protection. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue and trifluoroacetic acid (1 mL) were dissolved in dichloromethane (5 mL), and the mixture was reacted at room temperature for 1 hour. After the reaction was completed under LC-MS monitoring, the solution was concentrated under reduced pressure. The residue, compound II-5 (2 mg, 0.009 mmol), and N,N-diisopropylethylamine (30 μL, 0.182 mmol) were dissolved in dimethyl sulfoxide (3 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (3.15 mg, 0.008 mmol) was added, and the mixture was reacted at room temperature for 15 minutes. After the reaction was completed under LC-MS monitoring, the solution was purified by preparative liquid chromatography to give the title compound XZ1526C (8 mg, 21% yield, 3 steps). LC-MS (ESI): m / z 1291.2 [(M+2H) / 2] + .

[0599] Step 6. Synthesis of compounds XZ1539 and XZ1541.

[0600] Step 6.1. Synthesis of compound XZ1539. Compound IV-2 (20 mg, 0.015 mmol), compound II-9 (24 mg, 0.021 mmol), copper sulfate pentahydrate (0.73 mg, 0.003 mmol), and sodium ascorbate (1.16 mg, 0.006 mmol) were dissolved in 1,4-dioxane (3 mL), tert-butanol (2 mL), and water (1 mL). The mixture was reacted at 50 °C for 5 hours under nitrogen protection. After the reaction was monitored by LC-MS until complete, the mixture was concentrated under reduced pressure. The residue and trifluoroacetic acid (1 mL) were dissolved in dichloromethane (5 mL), and the mixture was reacted at room temperature for 1 hour. After the reaction was completed under LC-MS monitoring, the solution was concentrated under reduced pressure. The residue, compound IV-4 (3 mg, 0.009 mmol), and N,N-diisopropylethylamine (30 μL, 0.182 mmol) were dissolved in dimethyl sulfoxide (3 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (3.15 mg, 0.008 mmol) was added, and the mixture was reacted at room temperature for 15 minutes. After the reaction was completed under LC-MS monitoring, the solution was purified by preparative liquid chromatography to give the title compound XZ1539 (15 mg, 31% yield, 3 steps). 1H NMR(600MHz,DMSO-d6)δ11.88(s,1H),10.78(s,1H),10.02(d,J=13.9Hz,1H),9.12 (s,2H),8.17-8.11(m,1H),8.10-8.03(m,2H),7.92(d,J=8.6Hz,1H),7.81(d,J=19. 3Hz,1H),7.66-7.51(m,4H),7.26-7.16(m,2H),7.13-7.05(m,3H),6.88(dd,J=23.5 ,8.3Hz,2H),5.96(t,J=6.0Hz,1H),5.39(s,2H),5.02(s,2H),4.59-4.37(m,9H),4. 27-4.22(m,1H),4.00(d,J=28.6Hz,2H),3.86-3.71(m,3H),3.60(d,J=7.0Hz,3H), 3.53-3.40(m,104H),3.24(s,3H),3.10-2.94(m,9H),2.68-2.60(m,1H),2.55(t,J= 7.1Hz,2H),2.43-2.32(m,2H),2.18-2.09(m,1H),2.05-1.94(m,2H),1.88-1.78(m, 2H),1.70(s,2H),1.49-1.33(m,2H),0.85(dd,J=20.3,6.7Hz,6H).LC-MS(ESI):m / z 1318.7[(M+2H) / 2] + .

[0601] Step 6.2. Synthesis of compound XZ1541. Compound IV-2 (20 mg, 0.015 mmol), compound II-10 (24 mg, 0.021 mmol), copper sulfate pentahydrate (0.73 mg, 0.003 mmol), and sodium ascorbate (1.16 mg, 0.006 mmol) were dissolved in 1,4-dioxane (3 mL), tert-butanol (2 mL), and water (1 mL), and reacted at 50 °C for 1 hour under nitrogen protection. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure. The residue and trifluoroacetic acid (1 mL) were dissolved in dichloromethane (5 mL), and reacted at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure. The residue, compound IV-7 (3 mg, 0.009 mmol), and N,N-diisopropylethylamine (300 μL) were dissolved in dimethylformamide (3 mL), and reacted at room temperature for 3 hours. After the reaction was monitored to be complete by LC-MS, the title compound XZ1541 (11 mg, yield 27%, 3 steps) was purified by preparative liquid chromatography. 1H NMR(800MHz,DMSO-d6)δ11.87(s,1H),10.78(s,1H),10.02(d,J=19.0Hz,1H),9.11(s,2H) ,8.13(d,J=7.9Hz,1H),8.09-8.02(m,2H),7.92(d,J=8.6Hz,1H),7.81(d,J=26.0Hz,1H),7 .66-7.51(m,4H),7.26-7.16(m,2H),7.12-6.98(m,3H),6.88(dd,J=33.1,8.2Hz,2H),5.98 (s,2H),5.40(s,1H),5.02(s,2H),4.82-4.43(m,10H),4.40-4.36(m,1H),4.26-4.21(m,1H ),4.02(s,2H),3.82-3.78(m,2H),3.74(dd,J=11.2,4.9Hz,1H),3.62-3.58(m,6H),3.52-3 .45(m,101H),3.13-2.92(m,10H),2.56-2.53(m,2H),2.49-2.43(m,3H),2.43-2.38(m,1H) ,2.37-2.32(m,1H),2.18-2.09(m,1H),2.04-1.95(m,2H),1.87-1.78(m,2H),1.74-1.66(m ,1H),1.63-1.56(m,1H),1.48-1.33(m,2H),0.85(dd,J=26.6,6.7Hz,6H).LC-MS(ESI):m / z 1348.1[(M+2H) / 2] + .

[0602] Step 7. Synthesis of compounds XZ1533, XZ1537 and XZ1543.

[0603] Step 7.1. Synthesis of compound XZ1533. Using compounds XZ1717-C2 and III-4 as starting materials, the title compound XZ1533 was obtained via the same synthetic route as XZ1534. 1H NMR (400MHz, DMSO-d6) δ11.88(d,J=2.8Hz,1H),10.78(s,1H),10.00(d,J=9.5Hz,1H),8.14-8.02(m,3H),7.88-7.78(m,2H),7.67-7.4 9(m,4H),7.29-7.15(m,2H),7.12-7.06(m,2H),7.00(s,2H),6.95-6.84(m,2H),6.10-5.79(m,2H),5.02(s,2H),4.81-4.32(m,11H),4. 22(t,J=7.6Hz,1H),4.00(d,J=20.0Hz,2H),3.85-3.66(m,6H),3.63-3.52(m,37H),3.50-3.31(m,73H),3.14-2.90(m,9H),2.70-2.59 (m,1H),2.44(t,J=6.3Hz,3H),2.37-2.27(m,1H),2.22-1.89(m,3H),1.75-1.33(m,4H),0.83(dd,J=14.6,6.7Hz,6H).LC-MS(ESI):m / z 1320.6[(M+2H) / 2] + .

[0604] Step 7.2. Synthesis of compound XZ1537. Using compounds XZ1717-C2 and III-4 as starting materials, the title compound XZ1537 was obtained via the same synthetic route as XZ1534. LC-MS (ESI): m / z 1056.1 [(M+2H) / 2] + .

[0605] Step 7.3. Synthesis of compound XZ1543. Using compounds XZ1717-C2 and III-4' as starting materials, the title compound XZ1543 was obtained via the same synthetic route as XZ1534. 1H NMR (600MHz, DMSO-d6) δ11.87(s,1H),10.78(s,1H),9.98(d,J=15.0Hz,1H),8.09(d,J=7.3Hz,1H),8.04(s,1H),7.86-7.78(m ,3H),7.67-7.50(m,4H),7.26-7.16(m,2H),7.11-7.07(m,2H),7.00(s,2H),6.96-6.86(m,2H),6.05(s,2H),5.00(s,2H),4.8 2-3.52(m,80H),3.50-3.35(m,49H),3.14-2.85(m,11H),2.68-2.60(m,1H),2.49-2.41(m,4H),2.37-2.30(m,1H),2.21-2.10 (m,1H),2.07-1.92(m,2H),1.71(s,1H),1.65-1.57(m,1H),1.51-1.34(m,2H),0.83(dd,J=20.7,6.7Hz,6H).LC-MS(ESI):m / z 1327.2[(M+2H) / 2] + .

[0606] Step 8. Synthesis of compounds XZ1540, XZ1542 and XZ1543B.

[0607] Step 8.1. Synthesis of compound XZ1540. Compound IV-2 (20 mg, 0.015 mmol), compound II-10 (24 mg, 0.021 mmol), copper sulfate pentahydrate (0.73 mg, 0.003 mmol), and sodium ascorbate (1.16 mg, 0.006 mmol) were dissolved in 1,4-dioxane (3 mL), tert-butanol (2 mL), and water (1 mL), and reacted at 50 °C for 1 hour under nitrogen protection. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure. The residue and trifluoroacetic acid (1 mL) were dissolved in dichloromethane (5 mL), and reacted at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure. The residue, compound IV-5 (3 mg, 0.009 mmol), and N,N-diisopropylethylamine (300 μL) were dissolved in dimethylformamide (3 mL), and reacted at room temperature for 3 hours. After the reaction was monitored to be complete by LC-MS, the title compound XZ1541 (6 mg, yield 14%, 3 steps) was purified by preparative liquid chromatography. 1H NMR (600MHz, DMSO-d6) δ12.21(d,J=85.7Hz,1H),11.88(s,1H),10.78(s,1H),10.02(d,J=15.4Hz,1H),8.14-8.01(m,3H),7.85-7.78(m,2H),7.6 8-7.49(m,4H),7.33-7.15(m,12H),7.13-7.04(m,3H),6.94-6.82(m,2H) ,6.41(s,1H),5.97(s,1H),5.39(s,1H),5.00(d,J=19.4Hz,2H),4.64-4. 35(m,10H),4.24-4.18(m,1H),4.10-3.70(m,5H),3.62-3.58(m,4H),3. 53-3.43(m,100H),3.02(d,J=27.0Hz,10H),2.69-2.60(m,1H),2.46-2.4 2(m,2H),2.24-2.08(m,3H),2.05-1.92(m,2H),1.69(s,1H),1.65-1.31( m,8H),1.28-1.16(m,3H),0.83(dd,J=20.4,6.6Hz,6H).LC-MS(ESI):m / z 1427.6[(M+2H) / 2] + .

[0608] Step 8.2. Synthesis of compound XZ1542. Compound IV-2 (20 mg, 0.015 mmol), compound II-10 (24 mg, 0.021 mmol), copper sulfate pentahydrate (0.73 mg, 0.003 mmol), and sodium ascorbate (1.16 mg, 0.006 mmol) were dissolved in 1,4-dioxane (3 mL), tert-butanol (2 mL), and water (1 mL), and reacted at 50 °C for 1 hour under nitrogen protection. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure. The residue and trifluoroacetic acid (1 mL) were dissolved in dichloromethane (5 mL), and reacted at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure. The residue, compound IV-6 (3 mg, 0.009 mmol), and N,N-diisopropylethylamine (300 μL) were dissolved in dimethylformamide (3 mL), and reacted at room temperature for 3 hours. After the reaction was completed as monitored by LC-MS, the title compound XZ1541 (10 mg, yield 23%, 3 steps) was purified by preparative liquid chromatography. LC-MS (ESI): m / z 1428.2 [(M+2H) / 2] + .

[0609] Step 8.3. Synthesis of compound XZ1543B. Compound IV-2' (20 mg, 0.015 mmol), compound II-10 (24 mg, 0.021 mmol), copper sulfate pentahydrate (0.73 mg, 0.003 mmol), and sodium ascorbate (1.16 mg, 0.006 mmol) were dissolved in 1,4-dioxane (3 mL), tert-butanol (2 mL), and water (1 mL), and reacted at 50 °C for 1 hour under nitrogen protection. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure. The residue and trifluoroacetic acid (1 mL) were dissolved in dichloromethane (5 mL), and reacted at room temperature for 1 hour. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure. The residue, compound IV-6 (3 mg, 0.009 mmol), and N,N-diisopropylethylamine (300 μL) were dissolved in dimethylformamide (3 mL), and reacted at room temperature for 3 hours. After the reaction was monitored to be complete by LC-MS, the title compound XZ1541 (11 mg, yield 26%, 3 steps) was purified by preparative liquid chromatography. LC-MS (ESI): m / z 1435.6 [(M+2H) / 2] + .

[0610] Scheme 5. Synthesis of XZ1535 and XZ1536

[0611]

[0612] Step 1. Synthesis of compound V-1. Using compounds XZ1740-C2 and III-4 as starting materials, the title compound V-1 (60.0 mg, 66% yield) was obtained via the same synthetic route as XZ1525A. LC-MS (ESI): m / z 1357.5 [(M+2H) / 2] + .

[0613] Step 2. Synthesis of compound XZ1535. Compound V-1 (30 mg, 0.02 mmol), compound II-10 (52 mg, 0.05 mmol), copper sulfate pentahydrate (3 mg, 0.01 mmol), and sodium ascorbate (7 mg, 0.03 mmol) were dissolved in 1,4-dioxane (3 mL), tert-butanol (2 mL), and water (1 mL). The mixture was reacted at 50 °C for 1 hour under nitrogen protection. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure. The residue and trifluoroacetic acid (1 mL) were dissolved in dichloromethane (5 mL), and the mixture was reacted at room temperature for 30 minutes. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure. The residue, compound II-11 (7 mg, 0.03 mmol), and N,N-diisopropylethylamine (10 μL, 0.06 mmol) were dissolved in dimethyl sulfoxide (3 mL), and the mixture was reacted at room temperature for 20 minutes. After the reaction was completed as monitored by LC-MS, the title compound XZ1535 (13 mg, yield 21%, 3 steps) was purified by preparative liquid chromatography. LC-MS (ESI): m / z 1313.2 [(M+2H) / 2] + .

[0614] Step 3. Synthesis of compound V-2. Using compounds XZ1723B and III-4 as starting materials, the title compound V-2 (36.0 mg, 73% yield) was obtained via the same synthetic route as XZ1525A. LC-MS (ESI): m / z 1373.5 [M+H] + .

[0615] Step 4. Synthesis of compound XZ1536. Compound V-2 (35 mg, 0.03 mmol), compound II-10 (60 mg, 0.05 mmol), copper sulfate pentahydrate (6 mg, 0.03 mmol), and sodium ascorbate (10 mg, 0.05 mmol) were dissolved in 1,4-dioxane (3 mL), tert-butanol (2 mL), and water (1 mL). The reaction was carried out at 50 °C for 1 hour under nitrogen protection. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure. The residue and trifluoroacetic acid (1 mL) were dissolved in dichloromethane (5 mL), and the mixture was reacted at room temperature for 30 minutes. After the reaction was completed by LC-MS, the mixture was concentrated under reduced pressure. The residue, compound II-11 (4 mg, 0.05 mmol), and N,N-diisopropylethylamine (13 μL, 0.10 mmol) were dissolved in dimethyl sulfoxide (3 mL), and the mixture was reacted at room temperature for 20 minutes. After the reaction was completed as monitored by LC-MS, the title compound XZ1536 (14 mg, 20% yield, 3 steps) was purified by preparative liquid chromatography. LC-MS (ESI): m / z 1321.2 [(M+2H) / 2] + .

[0616] Example 3. Preparation and characterization of conjugates of novel degradation agent conjugates

[0617] Option 1. A preparation procedure for a coupling agent (lower DAR)

[0618] Antibody reduction: 5 mg / mL antibody solution was treated with 2.5 equivalents of tri-(2-carboxyethyl)phosphine (TCEP) in 10 mM PBS pH 7.4 buffer and incubated at 37 °C for 1 hour to reduce interchain disulfide bonds.

[0619] Antibody-drug conjugation: Add 6 equivalents of dimethyl sulfoxide (DMSO) solution of the linker-degrader conjugate to the above-described reducing antibody solution, and add an appropriate amount of DMSO to bring the final concentration of DMSO to 15% (v / v). Incubate the resulting reaction mixture at room temperature for 1 hour. Filter the resulting conjugate through a 0.20 μm PES membrane and concentrate it using an Amicon Millipore ultracentrifuge with a molecular weight cutoff of 30 kD.

[0620] Concentration was detected using Nanodrop at 280 nm.

[0621] Size exclusion chromatography (SEC) was used to determine the purity and aggregation degree of the conjugates: A 7.8 × 300 mm TSKGel 3000SWXL column (Tosoh Bioscience) with 5 μm particles was used. Elution was performed isocratically with a mobile phase of 0.3 M potassium dihydrogen phosphate (pH 6.5) and 15% (v / v) isopropanol at a rate of 1.0 mL / min for 30 min. The purity of the conjugates of the new degradation agent was determined based on the retention times and peak areas at 280 nm and 254 nm.

[0622] DAR was analyzed by reversed-phase liquid chromatography (RPLC): A 2.1 × 100 mm BioREsolve™ RP Polyphenyl column with 2.7 μm particles was used. Mobile phase A was 0.1% aqueous trifluoroacetic acid (TFA), and mobile phase B was an acetonitrile solution containing 0.1% TFA. The analyte was eluted with a linear gradient of 35%–80% B over 27 min at a flow rate of 0.2 mL / min and a column temperature of 80 °C. Detection was performed at 280 nm and 254 nm.

[0623] Hydrophobic interaction chromatography (HIC) analysis of DAR: A 4.6 × 100 mm TSKgelButyl-NPR column with 2.5 μm particles was used. Mobile phase A was an aqueous solution containing 1.5 mM ammonium sulfate and 25 mM disodium hydrogen phosphate at pH 7.0, and mobile phase B was an aqueous solution containing 20% ​​isopropanol and 25 mM disodium hydrogen phosphate at pH 7.0. The analyte was eluted with a linear gradient of 30%–70% B over 30 min at a flow rate of 0.5 mL / min and a column temperature of 25 °C. Detection was performed at 280 nm and 254 nm.

[0624] Sample purification: In vivo test samples can be purified using a 5 mL strong cation exchange chromatography column as needed. Mobile phase A is 20 mM PB solution at pH 6.5, and mobile phase B is 10 mM PBS buffer containing 1 M sodium chloride at pH 7.4. A gradient purification of the new degradation agent conjugate is then performed. After pooling, the samples are concentrated using an Amicon Millipore ultracentrifuge with a molecular weight cutoff of 30 kDa.

[0625] Option 2. A preparation procedure for a coupling compound (higher DAR)

[0626] Antibody reduction: 5 mg / mL antibody solution was treated with 10 equivalents of tri-(2-carboxyethyl)phosphine (TCEP) in 10 mM PBS pH 7.4 buffer and incubated at 37°C for 1 hour to reduce interchain disulfide bonds.

[0627] Antibody-drug conjugation: 12 equivalents of dimethyl sulfoxide (DMSO) solution of the linker-degrader conjugate were added to the above-described reducing antibody solution, and an appropriate amount of DMSO was added to bring the final concentration of DMSO to 15% (v / v). The resulting reaction mixture was allowed to stand at room temperature for 1 hour. The resulting new degrader conjugate was filtered through a 0.20 μm PES membrane and concentrated using an Amicon Millipore ultracentrifuge with a molecular weight cutoff of 30 kD.

[0628] Concentration was detected by Nanodrop at 280 nm, and the sample characterization and purification methods were the same as in Scheme 1.

[0629] Option 3. A preparation procedure for a coupling agent (fixed DAR)

[0630] Antibody reduction: 5 mg / mL antibody solution was treated with 10 equivalents of tri-(2-carboxyethyl)phosphine (TCEP) in 10 mM PBS pH 7.4 buffer and incubated at 37°C for 1 hour to reduce interchain disulfide bonds.

[0631] Antibody-drug conjugation: Add 6 equivalents of dimethyl sulfoxide (DMSO) solution of the linker-degrader conjugate to the above-described reducing antibody solution, and add an appropriate amount of DMSO to bring the final concentration of DMSO to 15% (v / v). Incubate the resulting reaction mixture at room temperature for 1 hour. Filter the resulting new degrader conjugate through a 0.20 μm PES membrane and concentrate it using an Amicon Millipore ultracentrifuge with a molecular weight cutoff of 30 kD.

[0632] Concentration was detected by Nanodrop at 280 nm, and the sample characterization and purification methods were the same as in Scheme 1.

[0633] Option 4. A preparation procedure for a coupling compound (low DAR)

[0634] Antibody reduction: 5 mg / mL antibody solution was treated with 1.5 equivalents of tri-(2-carboxyethyl)phosphine (TCEP) in 10 mM PBS pH 7.4 buffer and incubated at 37 °C for 1 hour to reduce interchain disulfide bonds.

[0635] Antibody-drug conjugation: Add 3 equivalents of dimethyl sulfoxide (DMSO) solution of the linker-degrader conjugate to the above-described reducing antibody solution, and add an appropriate amount of DMSO to bring the final concentration of DMSO to 15% (v / v). Incubate the resulting reaction mixture at room temperature for 1 hour. Filter the resulting new degrader conjugate through a 0.20 μm PES membrane and concentrate it using an Amicon Millipore ultracentrifuge with a molecular weight cutoff of 30 kD.

[0636] Concentrations were detected using Nanodrop at 280 nm. Sample characterization and purification methods were the same as in Scheme 1. Purification of specific DAR samples: If necessary, a 10 mL UniHR Butyl-30L hydrophobic chromatography column was used. Mobile phase A was an aqueous solution containing 1 M ammonium sulfate and 25 mM disodium hydrogen phosphate at pH 7.0, and mobile phase B was an aqueous solution containing 20% ​​isopropanol and 25 mM disodium hydrogen phosphate at pH 7.0. A linear gradient was employed to purify the new degradation agent conjugate. After pooling, the samples were concentrated using an Amicon ultracentrifuge (Millipore) with a molecular weight cutoff of 30 kD.

[0637] Table 1. Characterization of conjugates of the new degradation agent conjugates

[0638]

[0639]

[0640]

[0641] [a] Aggregation test: A: <5%; B: 5%-20%; C: >20%; [b] JMC12 is a GNE-987-derived conjugate (linker-degrader conjugate). The structures of JMC12 and its corresponding active degrader are shown below. JMC12 cell activity is significantly lower than that of GNE987. [c] The HIC method was used for analysis; [d] WO2022254377; [e] Molecular Cancer Therapeutics 23.10(2024):1367-1377.

[0642]

[0643] As shown in Table 1 above, the conjugates (linker-degrader conjugates) with specific structures in this application provide a significantly improved DAR. The amount of degrader in the antibody conjugates obtained using the same conjugation method (with the same equivalent amount of linker-degrader conjugate) is significantly increased compared to the conjugates in this application. Furthermore, most of the antibody conjugates in this application still exhibit low aggregation degree and higher physical stability even at high DAR.

[0644] Example 4. In vitro antitumor activity test of the new degradation agent

[0645] Human breast cancer cells BT-474 (Her2+) and MDA-MB-231 (triple-negative breast cancer) in the logarithmic growth phase were seeded into 96-well culture plates at an appropriate density of 100 μL per well. After overnight culture, different concentrations of drugs were added and treated for 3 days. Each concentration was set up in triplicate wells, and corresponding concentrations of solvent control and cell-free zeroing wells were also set up. After treatment, adherent cells were decanted from the culture medium and fixed with 10% (w / v) trichloroacetic acid (100 μL / well) at 4°C for 1 h. They were then washed five times with distilled water and dried at room temperature. 100 μL of SRB solution (Sigma, St. Louis, MO, USA) (4 mg / mL, dissolved in 1% glacial acetic acid) was added to each well, and the cells were incubated for staining at room temperature for 15 min. Unbound SRB was washed five times with 1% glacial acetic acid, and the cells were dried at room temperature. 150 μL of 10 mM Tris solution was added to each well, and the optical density (OD value) at 560 nm was measured using a SpectraMax 190 microplate reader. The inhibition rate of the compound on cell proliferation was calculated using the following formula: Inhibition rate (%) = [1 - (OD administration wells - OD negative control wells) / (OD positive control wells - OD negative control wells)] × 100%. The IC50 was estimated using the four-parameter method. 50 Each experiment was repeated three times independently, with three replicates for each concentration each time.

[0646] The results showed that the novel degrading agents disclosed herein exhibited certain inhibitory activity against the tested cell lines in vitro, and the inhibitory activity was significantly better than that of the existing representative BET degrading agent MZ-1. The inhibitory activity of several novel degrading agents was comparable to that of the potent BET degrading agent GNE-987 (Table 2).

[0647] Table 2. In vitro antitumor activity of the new degradation agent

[0648]

[0649]

[0650] [a] A: IC 50 <10nM; B: 10nM <IC 50 <100nM; C: 100nM <IC 50 <300nM; D:IC 50 >300nM; ND: Not detected.

[0651] Example 5. Protein degradation activity of the new degrading agent

[0652] Human hepatocellular carcinoma cells (Huh-7) in logarithmic growth phase were seeded in six-well plates and cultured overnight. After treatment with different concentrations of compounds for 16 hours, cells were collected. Cells were washed once with pre-cooled PBS and lysed with 1×SDS loading buffer (50mM Tris, pH 6.8, 100mM DTT, 2% SDS, 0.1% bromophenol blue, 10% glycerol). Cell lysates were collected, heated in a boiling water bath for 10 min, and then centrifuged at 12000 rpm for 5 min at 4°C. The supernatant was used for SDS-PAGE electrophoresis. After electrophoresis, proteins were transferred to nitrocellulose membranes using a rapid wet transfer apparatus. The target bands were blocked for 1 h at room temperature in a blocking buffer containing 5% skim milk powder (5% skim milk powder, 20mM Tris-HCl, pH 7.2-7.4, 150mM NaCl, 0.1% Tween-20). The membranes were then incubated overnight at 4°C with the corresponding primary antibody. Wash three times at room temperature for 10 min each time with TBST washing buffer (100 mM Tris-HCl pH 7.2-7.4, 0.9% NaCl, 0.2% Tween-20). Add horseradish peroxidase-labeled secondary antibody and incubate on a shaker at room temperature for 1 h. Wash three more times with TBST for 10 min each time, then develop color and expose to light. BRD4 (#13440) was purchased from Abconal; Actin (#60008-1-Ig) was purchased from Proteintech; horseradish peroxidase-labeled goat anti-mouse secondary antibody (#401215) and horseradish peroxidase-labeled goat anti-rabbit secondary antibody (#401353) were purchased from Calbiochem.

[0653] Test results show that the representative novel degrading agents of this disclosure, after co-incubation with cells for 16 hours, can selectively degrade intracellular BRD4 in a dose-dependent manner. Figure 2 Among them, XZ1757A and XZ1757B also showed significant BRD4 protein degradation effects at low concentrations (0.08 nM), and the half-maximal concentration (DC) of degradation was also high. 50 The levels were significantly lower than those of the control compound GNE-987 (see [link]). Figure 2 ).

[0654] Example 6. In vitro antitumor activity of conjugates of novel degradation agent conjugates

[0655] Human breast cancer cells BT-474 (Her2+), JIMT-1 (Her2+, B7H3+), MDA-MB-231 (triple-negative breast cancer, B7H3+), human liver cancer cells PLC-PRF-5 (B7H3+), human leukemia cells CCRF-CEM (B7H3-), human non-small cell lung cancer cells HCC827 (EGFR+), and NCI-H1581 (EGFR-) in logarithmic growth phase were seeded at appropriate densities into 96-well plates, 150 μL per well. After overnight culture, different concentrations of the drug were added for 6 days of treatment, with each concentration in triplicate. Corresponding solvent controls and cell-free nulling wells were also included. After treatment, the viability of BT-474, JIMT-1, MDA-MB-231, and PLC-PRF-5 cells was assessed using the SRB assay. The culture medium was immediately discarded, and 10% (w / v) trichloroacetic acid (100 μL / well) was added for fixation at 4°C for 1 h. The cells were then washed five times with distilled water and dried at room temperature. 100 μL of SRB solution (Sigma, St. Louis, MO, USA) (4 mg / mL, dissolved in 1% glacial acetic acid) was added to each well, and the cells were incubated for staining at room temperature for 15 min. Unbound SRB was washed five times with 1% glacial acetic acid, and the cells were dried at room temperature. 150 μL of 10 mM Tris solution was added to each well, and the optical density (OD value) at 560 nm was measured using a SpectraMax 190 microplate reader. CCRF-CEM, NCI-H1581, and HCC827 cells were assessed for cell proliferation inhibition using CCK8 assay. After the treatment with the test compound, 10 μL of CCK-8 reagent was added to each well, and the wells were incubated at 37°C for 2-4 hours. The optical density (OD value) at 450 nm was then measured using a SpectraMax 190 microplate reader. The inhibition rate of the compound on cell proliferation was calculated using the following formula: Inhibition rate (%) = [1 - (OD administration wells - OD negative control wells) / (OD positive control wells - OD negative control wells)] × 100%. The IC50 was estimated using the four-parameter method. 50Each experiment was repeated three times independently, with three replicates for each concentration each time.

[0656] The results showed that the novel degradation agent conjugate conjugates disclosed herein exhibited certain inhibitory activity against the tested cell lines in vitro, and the inhibitory activity showed significant receptor-related selectivity (Tables 3-6). For example, the IC50 of the HER2-overexpressing cell lines BT-474 or JIMT-1 was significantly increased. 50 The inhibitory effect was far superior to that on the proliferation of human triple-negative breast cancer cells MDA-MB-231, with most conjugates showing orders of magnitude difference (Table 3).

[0657] Table 3. In vitro antitumor activity of HER2-new degradation agent conjugates

[0658]

[0659]

[0660] [a] Data source: https: / / depmap.org / portal / , log2(TPM+1); [b] A: IC 50 <10nM; B: 10nM <IC 50 <100nM; C: 100nM <IC 50 <300nM; D:IC 50 >300nM; ND: Not detected.

[0661] Table 4. In vitro antitumor activity of B7H3-new degradation agent conjugates

[0662]

[0663] [a] Data source: https: / / depmap.org / portal / , log2(TPM+1); [b] A: IC 50 <10nM; B: 10nM <IC 50 <100nM; C: 100nM <IC 50 <300nM; D:IC 50 >300nM; ND: Not detected.

[0664] Table 5. In vitro antitumor activity of EGFR-new degradation agent conjugates

[0665]

[0666] [a]Data source: https: / / depmap.org / portal / , log2(TPM+1)

[0667] [b] A: IC 50 <10nM; B: 10nM <IC 50 <100nM; C: 100nM <IC 50 <300nM; D:IC 50 >300nM.

[0668] Table 6. In vitro antitumor activity of Trop2-new degradation agent conjugates

[0669]

[0670]

[0671] [a] Data source: https: / / depmap.org / portal / , log2(TPM+1); [b] A: IC 50 <10nM; B: 10nM <IC 50 <100nM; C: 100nM <IC 50 <300nM; D:IC 50 >300nM; ND: Not detected.

[0672] Example 7. In vitro protein degradation activity of conjugates of novel degradation agent conjugates

[0673] Taking novel HER2-targeting degradative conjugates as an example, the degradation ability of these conjugates on the target protein was investigated in cell lines with significant differences in receptor expression. Human breast cancer cells in logarithmic growth phase, MDA-MB-231 (Her2-), BT-474 (Her2+), and JIMT-1 (Her2+), were seeded in six-well plates and cultured overnight before being added to… Figure 3Cells were treated with different concentrations of compounds XZ1522A-b, XZ1522B-a, XZ1522C-a, and GNE-987 for 24 hours, and then collected. Cells were washed once with pre-cooled PBS, and then lysed with 1×SDS loading buffer (50mM Tris pH 6.8, 100mM DTT, 2% SDS, 0.1% bromophenol blue, 10% glycerol). Cell lysates were collected, heated in a boiling water bath for 10 min, and then centrifuged at 12000 rpm for 5 min at 4℃. The supernatant was used for SDS-PAGE electrophoresis. After electrophoresis, proteins were transferred to a nitrocellulose membrane using a rapid wet transfer apparatus. After transfer, Ponceau S staining was used to confirm the transfer status and the position of the protein bands on the nitrocellulose membrane. After labeling, the target band was blocked for 1 hour at room temperature on a shaker with blocking buffer containing 5% skim milk (5% skim milk, 20 mM Tris-HCl, pH 7.2-7.4, 150 mM NaCl, 0.1% Tween-20). Then, the membrane was incubated overnight at 4°C with primary antibody. It was washed three times at room temperature for 10 minutes each with TBST washing buffer (100 mM Tris-HCl, pH 7.2-7.4, 0.9% NaCl, 0.2% Tween-20). Horseradish peroxidase-labeled secondary antibody was added, and the membrane was incubated for 1 hour at room temperature on a shaker. After washing three more times with TBST for 10 minutes each, the membrane was developed and exposed to sunlight.

[0674] Western blot results showed that, similar to GNE-987, the three novel HER2-degrading conjugates we detected significantly degraded BRD4 in a concentration-dependent manner and downregulated the expression of its downstream key signal c-Myc. Moreover, this degradation effect was more pronounced in Her2+ human breast cancer cells BT-474 and JIMT-1 cells. Figure 3 ).

[0675] Example 8. In vivo antitumor activity of conjugates of degradation agents

[0676] We used the Her2+ human breast cancer JIMT-1 subcutaneous xenograft model to investigate the in vivo activity of Her2-degrading agent conjugates. These new degrading agent conjugates also showed good in vivo antitumor activity.

[0677] Under aseptic conditions, tumor tissue in its vigorous growth phase was cut into 1.5 mm pieces. 3 Subcutaneous xenografts were inoculated into the right axilla of SCID mice. The diameter of the subcutaneous xenografts in SCID mice was measured using calipers. The xenografts were allowed to grow to an average volume of approximately 150 mm². 3Animals were randomly divided into groups. Each treatment group received a single intravenous injection via tail vein on the day of grouping (d0), while the solvent control group received an equal volume of PBS solution. Throughout the experiment, tumor diameter was measured twice weekly, and mouse weight was also measured. Tumor volume (TV) was calculated using the formula: TV = 1 / 2 × a × b 2 Where a and b represent length and width, respectively. The relative tumor volume (RTV) is calculated based on the measurement results using the formula: RTV = V t / V0. Where V0 is the tumor volume measured at the time of administration (d0), and Vt is the tumor volume at each measurement. The tumor growth inhibition rate (%) is used as the evaluation index of antitumor activity, calculated using the following formula: Tumor growth inhibition rate (%) = 100 - (T) RTV / C RTV )×100%, T RTV Treatment group RTV; C RTV Negative control group RT, TVt represents tumor volume measured at each time in the treatment group. Results are shown in Table 7 (tumor growth inhibition rate on day 14). Positive control T-DXd (5 mg / kg), as reported in the literature, significantly inhibited the growth of JIMT-1 subcutaneous xenografts with a single intravenous injection. In this experiment, each test compound also showed varying degrees of inhibitory effect on tumor growth, and this effect was dose-dependent (Table 7).

[0678] Table 7. In vivo antitumor activity of the new degradation agent conjugate conjugates

[0679]

[0680] [a] In all treatment groups with tumor growth inhibition rates greater than 50%, the t-test showed a significant difference compared to the control group.

[0681] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A protein-binding conjugate or a pharmaceutically acceptable salt thereof, characterized in that, The protein-binding conjugate is shown in Formula (I); in, a is a value between 1 and 8; L 1 For the connector part; L P It can be a cleavable or incuttable linking group; L 2 Groups that are absent or contain one or more PEG unit structures and / or polysarcosine unit structures; A is R E3 -R Linker -; where R E3 For the E3 ligase ligand moiety, R Linker For use with or without the E3 ligase ligand moiety and the BET family protein binding moiety as shown in Formula IA; L 3 Selected from the following group: amide, carbonyl, C1-8 alkylene amide, C1-8 alkylene carbonyl and C1-15 alkylene; Ar is an aromatic ring or a heteroaromatic ring; Bm is the binding moiety that can specifically bind to the target protein.

2. The protein-binding conjugate of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The protein-binding conjugate has one or more of the following characteristics: (a)L 1 -M Bm -L 1a -;in, M Bm Selected from the following group: The connection point for Bm; L 1a It is a chain-like linking group; (b)L P -M P2 -M P1 -M P3 -; where M P1 It is a polypeptide fragment; M P2 and M P3 Each independently is either non-NHCO-C(R) a )R b -CO-; where R a and R b Each is independently H or C1-4 alkyl; or R a and R b Together with the carbon atoms attached to them, they form C4-6 cycloalkyl groups; (c)L 2 In this context, the group containing one or more PEG unit structures and / or polysarcosine unit structures is -M. Ph -M 2L -M 2a -(CH2CH2O) q -M 2b or -M Ph -M 2L -M 2a -[CH2N(Me)CO] q -M 2b ;in, M Ph Selected from the following group: none, C1-6 alkylene, 1 to 6 heteroalkylene; M 2L Selected from the following group: none, -NHCO-, -N(C1-4 alkyl)CO-, linking groups formed by click chemistry; M 2a It is either non-alkyl or C1-4 alkylene; M 2b Selected from the group consisting of: none, C1-2 alkyl, C1-2 alkylene-acid groups; and q is an integer between 1 and 50; (d)L 3 Selected from the following groups: -NHCO-, -(CH2) m - and -CO-; where m is an integer from 1 to 12; (e)R Linker For none or as shown in the general formulas selected from Table 4.1a; Table A4.1a in, For The connection point of the methylene group; n is an integer from 0 to 10; W is non-native, carbonyl, C1-3 alkylene, or C1-3 alkylene-carbonyl; W L It is free of, NH or N (C1-3 alkyl); R L1 and R L2 Each is independently selected from the following group: H, C1-3 alkyl, C3-6 cycloalkyl, or R. L1 and R L2 And together with the carbon atoms connected to them, they form C3-6 cycloalkyl groups; W R It is a C3-10 cycloalkylene group or a 4- to 10-membered heteroalkylene group; (f)R E3 As shown in the general formula selected from Table 4.2a; Table A4.2a in, For The junction of the middle methylene group; X is either N or CH; Ring B is an optionally substituted C3-20 cycloalkyl or an optionally substituted 4- to 20-membered heterocyclic alkyl; The optional substitution refers to a group that is unsubstituted or one or more H atoms in the group being substituted by R atoms; R is selected from the following group: hydroxyl, amino, C1-4 alkyl; (g)Ar is a C6-10 aromatic ring or a 5- to 10-membered heteroaryl ring; (h) The binding portion is selected from the group consisting of: antibodies, antibody fragments of the antibody, or antigen-binding fragments of the antibody; (i) The target protein is a surface antigen.

3. The protein-binding conjugate as described in claim 2, or a pharmaceutically acceptable salt thereof, characterized in that, The protein-binding conjugate has one or more of the following characteristics: (a)L 1 -M Bm -M 1b -(M 1a ) p -M LP -;in, M Bm As defined in claim 2; M 1a Selected from the following group: C1-3 alkylene, C1-2 alkylene-O (preferably -CH2CH2O-); M 1b and M LP Each is independently selected from the group consisting of: none, C1-6 alkylene, C1-4 alkylene-CONH, C1-4 alkylene-CONH-C1-4 alkylene, C1-4 alkylene-NHCO, C1-2 alkylene-NHCO-C1-2 alkylene, C1-4 alkylene-CO, C1-4 alkylene-CO-C1-4 alkylene, C1-4 alkylene-NH, and C1-2 alkylene-NH-C1-2 alkylene; and p is an integer from 0 to 10; (b) Ring B is as shown in equation IB-1, IB-2 or IB-3: in, X 1 X 2 X 3 and X 4 Each independently as X a X a -X b or X a -X b -X c X 5 and X 6 Each independently is C(R) 1 ) or N; X 7 None (single key), X a or X a -X b ;and X a X b and X c Each is independently selected from the following group: C(R) 1 )2、N(R 1 ), O and S; R 1 Each can be H or R independently; (c)L P for Among them, the better, To be with L 1 The connection point; Z is the connection point with the aniline moiety. 1 and Z 2 Each is independently an empty or amino acid residue, Z 3 and Z 4 Each is an amino acid residue.

4. The protein-binding conjugate of claim 3 or a pharmaceutically acceptable salt thereof, characterized in that, The protein-binding conjugate has one or more of the following characteristics: (a) Ring B is a ring as shown in equation IA-1 or IA-2. (b)Z 1 For the absence of glycine residues; and / or, Z 2 The following amino acid residues are absent or selected: L-glutamine residue, D-glutamine residue, L-glutamate residue, D-glutamate residue, L-aspartic acid residue, D-aspartic acid residue, L-alanine residue, D-alanine residue, and glycine residue; and / or, Z 3 Selected from the group consisting of: L-valine residues, D-valine residues, L-alanine residues, D-alanine residues, L-phenylalanine residues, D-phenylalanine residues, and glycine residues; and / or, Z 4 Selected from the following group: L-alanine residue, D-alanine residue, L-citrulline residue, D-citrulline residue, L-asparagine residue, D-asparagine residue, L-lysine residue, D-lysine residue, L-phenylalanine residue, D-phenylalanine residue, and glycine residue.

5. The protein-binding conjugate of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The surface antigens include: 5T4, ACE, ADRB3, AKAP-4, ALK, androgen receptor, AOC3, APP, axonin 1, AXL, B7H3, B7-H4, BCL2, BCMA, bcr-abl, BORIS, BST2, C242, C4.4a, and CA. 125, CA6, CA9, CAIX, CCL11, CCR5, CD123, CD133, CD138, CD142, CD15, CD15-3, CD171, CD179a, CD18, CD19, CD19-9, CD2, CD20, CD22, CD23, CD24, CD25, CD27L, CD28, CD3, CD30, CD31, CD300LF, CD33, CD352, CD37, CD38, CD4, CD40, CD41, CD44, CD44v6, CD5 CD51, CD52, CD54, CD56, CD62E, CD62P, CD62L, CD70, CD71, CD72, CD74, CD79a, CD79b, CD80, CD90, CD97, CD125, CD138, CD141, CD147, CD152, CD154, CD326, CEA, CEACAM5, CFTR, condensation factor, cKit, closure protein 3, closure protein 18.2, CLDN6, CLEC12A, CLL-1, cll3, c-MET, Crypto 1. Growth factors, CS1, CTLA-4, CXCR2, CXORF61, cyclin B1, CYP1B1, cadherin-3, cadherin-6, DLL3, E7, EDNRB, EFNA4, EGFR, EGFRvIII, ELF2M, EMR2, ENPP3, EPCAM, EphA2, liver glycoside A4, liver glycoside B2, EPHB4, ERBB2 (Her2 / neu), ErbB3, ERG (TMPRSS2) ETS fusion gene), ETBR, ETV6-AML, FAP, FCAR, FCRL5, FGFR1, FGFR2, FGFR3, FGFR4, FLT3, folate receptor α, folate receptor β, FOLR1, Fos-associated antigen 1, fucose GM1, GCC, GD2, GD3, GloboH, GM3, GPC1, GPC2, GPC3, gplOO, GPNMB, GPR20, GPRC5D, GUCY2C, HAVCR1, HER2, HER3, HGF, HMI.

24. HMWMAA, HPV E6, hTERT, human telomerase reverse transcriptase, ICAM, ICOS-L, IFN-α, IFN-γ, IGF-I receptor, IGLL1, IL-2 receptor, IL-4 receptor, IL-13Ra2, IL-11Ra, IL-1, IL-12, IL-23, IL-13, IL-22, IL-4, IL-5, IL-6, interferon receptor, integrins (including α4, α). v β3, α v β5, α v β6, α1β4, α4β1, α4β7, α5β1, α6β4, α IIb β3 integrin), integrin αV, intestinal carboxyl esterase, KIT, LAGE-1a, LAIR1, LAMP-1, LCK, podin, LewisY, LFA-1 (CD11a), L-selectin (CD62L), LILRA2, LIV-1, LMP2, LRRC15, LY6E, LY6K, LY75, MAD-CT-1, MAD-CT-2, MAGE A1, MelanA / MART1, mesothelin, MLIAP, MSLN, mucin, MUC1, MUC16, mut hsp70-2, MYCN, myostatin, NA17, NaPi2b, NCA-90, NCAM, connexin-4, NGF, NOTCH1, NOTCH2, NOTCH3, NOTCH4, NY-BR-1, NY-ESO-1, o-acetyl-GD2, OR51E2, OY-TES1, p53, p53 mutant, PANX3, PAP, PAX3, PAX5, p-CAD, PCTA-1 / galactagoguerin 8, PD-L1, PD-L2, PDGFR, PDGFR-β, phosphatidylserine, PIK3CA, PLAC1, polysialic acid, prostate enzymes, prostate cancer cells, prostaglandins, copper *Pseudomonas aeruginosa*, rabies virus, survivin and telomerase, PRSS21, PSCA, PSMA, PTK7, RAGE-1, RANKL, Ras mutant, respiratory syncytial virus, rhesus monkey factor, RhoC, RON, ROR1, ROR2, RU1, RU2, sarcoma translocation breakpoint, SART3, SLAMF7, SLC44A4, sLe, SLITRK6, spermin 17, sphingosine 1-phosphate, SSEA-4, SSX2, STEAP1, TAG72, TARP, TCRβ, TEM1 / CD248, TEM7R, tendinin C, TF, TGF-1, TGF-β2, TNF-α, TGS5, Tie 2. TIM-1, TnAg, TRAC, TRAIL-R1, TRAIL-R2, TROP-2, TRP-2, TRPV1, TSHR, tumor antigen CTAA16.88, tyrosinase, UPK2, VEGF, VEGFR1, VEGFR2, vimentin, WT1, XAGE1, or combinations thereof.

6. The protein-binding conjugate of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The protein-binding conjugate has one or more of the following characteristics: (a)L 1 Selected from Table A1 below: Table A1 Where p is an integer from 1 to 10; For L P The connection point; The connection point for Bm; (b)L P Selected from Table A2 below; Table A2 in, To be with L 1 The connection point; and This is the connection point with the aniline portion; (c)L 2 Selected from Table A3 below Table A3 Where q is an integer between 1 and 50, and q' is an integer between 1 and 6; (d)A is as shown in the general formula selected from Table A5a below; Table A5a in, To be with L 3 The connection point; For The junction of the middle methylene group; X is either N or CH; n is an integer between 0 and 10; W is a carbonyl group or a C1-3 alkylene group; and Cycle B is an optionally substituted C3-C20 cycloalkyl or an optionally substituted 4- to 20-membered heterocyclic alkyl; The optional substitution refers to a group that is unsubstituted or one or more H atoms in the group being substituted by R atoms; R is selected from the following group: hydroxyl, amino, C1-4 alkyl; (e)Ar is a phenyl or a 5- or 6-membered nitrogen-containing heteroaromatic ring; (f) Bm is an antibody or its antibody fragment or its antigen-binding fragment selected from the following group: trastuzumab, pertuzumab, decituzumab, datopotamab, sacituzumab, vobramitamab, ifinatamab, depatuxizumab, cetuximab, panitumumab.

7. The protein-binding conjugate of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The protein-binding conjugates mentioned are selected from Table B; Table B: Protein Conjugates 8. A pharmaceutical composition, characterized in that, It comprises (i) the protein binding conjugate as claimed in claim 1 or a pharmaceutically acceptable salt thereof, and (ii) one or more pharmaceutically acceptable carriers.

9. Use of a protein binding conjugate as described in claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cancer.

10. The use as described in claim 1, characterized in that, The cancers mentioned are selected from the following group: breast cancer, lung cancer, stomach cancer, hepatocellular carcinoma, lymphoma, leukemia, pancreatic cancer, head and neck cancer, squamous cell carcinoma, urethral cancer, colorectal cancer, prostate cancer, ovarian cancer, bladder cancer, gastrointestinal stromal tumor, cervical cancer, esophageal cancer, peritoneal cancer, liver cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, vulvar cancer, thyroid cancer, penile cancer, plasmacytoma, myeloma, sarcoma, or combinations thereof.

11. The use as described in claim 1, characterized in that, The cancer treatment drug can be used in combination with an adjuvant; and the adjuvant is a cytotoxic agent or an immune response modulator.

12. A degrading agent or a pharmaceutically acceptable salt thereof, characterized in that, The degrading agent is shown in formula (II); in, A i For R i E3 -R i Linker -; where R i E3 For the E3 ligase ligand moiety, R i Linker For use with or without the E3 ligase ligand moiety and the BET family protein binding moiety as shown in Formula IA; Ar and L 3 The definition is the same as that in equation (I).

13. The degrading agent as described in claim 12, or a pharmaceutically acceptable salt thereof, characterized in that, The degrading agents are selected from Table C below; Table C: Degrading Agents 14. A connector-degrader coupling compound or a pharmaceutically acceptable salt thereof, characterized in that, The connector-degrader coupling is shown in formula (III); Among them, Ar, L 3 A and L 2 As defined in equation (I); L ii Bm It is a linker that has a binding moiety that can be further coupled to specifically bind to the target protein.

15. The connector-degrader conjugate as claimed in claim 14, or a pharmaceutically acceptable salt thereof, characterized in that, The connector-degrader couplings are selected from Table D below. Table D: Connector-Degrader Couplings 16. Use of the linker-degrader conjugate of claim 14 in the preparation of the protein binding conjugate of claim 1 or a pharmaceutically acceptable salt thereof.

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