Antibody-drug conjugate with connection system

Abstract

Provided are antibody-drug conjugates (ADCs) with linking systems, preparation methods therefor, and uses thereof. These linking systems have high linking stability and enhanced solubility, allowing for efficient conjugation of hydrophobic drugs and effective intracellular delivery of drugs.

Description

[0001] The present invention claims priority to the following six patent applications filed with the China National Intellectual Property Administration: Patent Application No. 2024101754438, filed on February 7, 2024, Patent Application No. 2024105358331, filed on April 30, 2024, Patent Application No. 2024107726728, filed on June 14, 2024, Patent Application No. 202411432986X, filed on October 14, 2024, Patent Application No. 2024115803285, filed on November 6, 2024, and Patent Application No. 2025101207110, filed on January 24, 2025, all entitled "ANTIBODY-DRUG CONJUGATE WITH CONNECTION SYSTEM", and these earlier-filed applications are incorporated herein by reference in their entirety. TECHNICAL FIELD

[0002] The present invention belongs to the field of medicinal chemistry, and specifically relates to antibody-drug conjugates with linking systems. BACKGROUND

[0003] Antibody-drug conjugates (ADCs) can selectively target cancer cells, thereby killing them with minimal impact on normal cells, ushering in a new era of cancer treatment. Several antibody-drug conjugates (ADCs) have been approved for marketing by FDA. Examples include Mylotarg, a CD33 antibody conjugated to calicheamicin; Adcetris, a CD30 antibody conjugated to MMAE, indicated for treating patients with Hodgkin lymphoma and anaplastic large cell lymphoma; Kadcyla, a Her2 antibody conjugated to a maytansine derivative, indicated for treating patients with HER2-positive breast cancer; DS8201, a Her2 antibody conjugated to the camptothecin derivative Dxd, indicated for treating patients with HER2-positive breast cancer; and Sacituzumab govitecan, targeting the TROP-2 antigen, indicated for treating triple-negative breast cancer.

[0004] According to the classic ADC drug mechanism of action, antibody-drug conjugates can specifically bind to cell surface proteins, and the resulting conjugates are endocytosed by the cells, thereby achieving the effect of targeted delivery of drug molecules to tumor cells. Therefore, the concentration of intracellular drugs is directly related to the distribution density of target sites on the cell surface that can be specifically recognized by antibodies. However, the density of target sites that can be recognized by antibodies on the molecular surface is usually low, resulting in low drug concentrations within the target cells. To address this issue, a common approach is to increase the DAR value of the ADC, thereby increasing the amount of active drug entering the cell and thus improving efficacy. However, according to the study by Hamblett et al. (Clin Cancer Res. 2004,10, 7063), increasing the DAR value of ADC usually affects the pharmacokinetic properties of ADC in vivo. ADC molecules will aggregate, plasma stability will decrease, and small molecule toxins in the blood will be released more, causing toxic side effects.

[0005] Since 2013, Daiichi Sankyo has filed a number of patent applications (CN201380053256.2, CN201910768778.X, CN201980061665.4, etc.), disclosing a series of antibody-drug conjugates with specific linker-toxin structures, and specifically disclosing ADCs with the following typical structures that exhibit superior performance. However, the stability of the linker needs to be further improved to further expand the safety window of the ADC. \ 0

[0006] Therefore, developing a linker-toxin structure that can be stably released in plasma and specifically released in tumor cells, and then developing new ADCs to improve the safety window of antibody-drug conjugates in clinical use, remains of great significance. SUMMARY OF THE INVENTION

[0007] The present invention provides a compound represented by formula (Ia) or (Ib), a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof: M-Z1-Tr1-L1-D (Ia) or M-Z1-L2(Tr2)-D (Ib) wherein M is a linker site to an antibody or an antigen-binding fragment thereof; preferably, M comprises a thiol-reactive group, an amino-reactive group, a carboxyl-reactive group, a proline residue-reactive group, a tyrosine residue-reactive group, a disulfide bond-bridging group, etc.; for an antibody incorporating an unnatural amino acid, M also comprises a bioorthogonal reactive group; preferably a thiol-reactive group, such as a methylsulfonylpyrimidine group, a methylsulfonylpyridine group, and a maleimide group, wherein the pyrimidine group, pyridine group, and maleimide group are optionally substituted with one or more substituents selected from the group consisting of halogen, CN, C1-6 alkyl, C1-6 alkoxy, C1-6 alkylene-O-C1-6 alkyl, and -O-(CH2CH2O)n1-CH3; n1 is an integer from 1 to 36; Z1 is a chemical bond, a divalent group, or a trivalent group substituted with a hydrophilic group, such as NH or -(CH2)qCONH-; q is an integer from 0 to 6; Tr1 is a divalent trigger group, preferably a peptide residue or a modified peptide residue, wherein the peptide residue or modified peptide residue comprises an optionally substituted natural or unnatural amino acid, an L- or D-amino acid; and it is selected from glycine-glycine-phenylalanine-glycine (GGFG), glutamic acid-valine-citrulline (EVC), valine-citrulline (VC), valine-alanine (VA), aspartic acid-valine-citrulline (DVC), glutamic acid-glycine-glycine-phenylalanine-glycine (EGGFG), aspartic acid-glycine-glycine-phenylalanine-glycine (DGGFG), and lysine-glycine-glycine-phenylalanine-glycine (KGGFG); L1 is a linking moiety between Tr1 and a bioactive molecule structural fragment D; preferably a chemical bond, -NH-Ci-6 alkyl-, -N(CH3)-Ci-6 alkyl-, h             , h L1 is further preferably -NH-CH2- or D is selected from bioactive molecule structural fragments; L2 is a trivalent linking moiety connecting Z1 or M, Tr2, and the bioactive molecule structural fragment D; Tr2 is a monovalent trigger group, preferably a peptide residue, a p—glucuronide group, or a p— galactoside group; in formula (Ia) or formula (Ib), M or Z1 has at least one monovalent hydrophilic group Hp1 substituent; and / or Z1 has at least one divalent hydrophilic group Hp2 inserted or substituted, the monovalent hydrophilic group or divalent hydrophilic group is linked to other moieties of formula (Ia) or formula (Ib) via any chemical bond or linking group; the monovalent hydrophilic group Hp1 is selected from: (1) monovalent polyethylene glycol groups, preferably: -(CH2CH2O)n1-T1, -(CH2CH2O(CH2)m1CONH)n1-T1, or cyclic groups comprising -(CH2CH2O)- segments, the cyclic group optionally comprising 1 to 3 heteroatoms such as N; (2) monovalent polysarcosines: -(N(CH3)CH2CO)n2-T2; (3) monovalent polybetaines: -(CH2CH2N(CH3)(CH2COOH))n3-T3 (inner salt form: -(CH2CH2N+(CH3)(CH2COO-))n3-T3), or -(CH2CH2N(CH3)(CH2SO3H))n3-T3; (4) groups comprising carboxyl groups: -(CH2)m2-X-(CH2)m3-COOH; (5) the combinations -Hp2-Hp1 of (1)-(4) above with the following divalent hydrophilic groups are also regarded as monovalent hydrophilic groups in the present invention; the divalent hydrophilic group Hp2 is selected from divalent groups derived by removing the capping group from the above monovalent hydrophilic groups, or n4, n5 or n6 repeating units of -K(Hp1)-, preferably: (1) divalent polyethylene glycol groups, preferably: -(CH2CH2O)n1-, -(CH2CH2O(CH2)m1CONH)n1-, -[K-(CH2CH2O)n1-T1]n4-, -[K-(CH2CH2O(CH2)m1CONH)n1- T1]n4-, or cyclic groups comprising -(CH2CH2O)- segments, the cyclic group optionally comprising 1 to 3 heteroatoms such as N, for example: (CH2CH2O)n (CH2CH2O)n H2            wherein n1’ and n1’’ are each an integer from 1 to 36, and q1 and q2 are each an integer selected from 1, 2, or 3; (2) divalent polysarcosines: -(N(CH3)CH2CO)n2- or -[K-(N(CH3)CH2CO)n2-T2]n5-; (3) divalent polybetaines: -(CH2CH2N(CH3)(CH2COOH))n3- or -[K-(CH2)m4N(CH3)2CH2COOH]n6-, or -(CH2CH2N(CH3)(CH2SO3H))n3- or -[K-(CH2)m4N(CH3)2CH2SO3H]n6-; (4) any combination of (1)-(3) above; wherein n1-n6 are each an integer from 1 to 36; m1-m5 are each an integer from 1 to 10; X is O, S or NH; K is any trivalent group, preferably a trivalent group derived from an amino acid; T1, T2 or T3 is selected from any capping group, preferably H, OH, C1-6 alkyl, carboxyl, -(CH2)m5-CONH-(CH2)m4N(CH3)2CH2COOH, -(CH2)m4N(CH3)2CH2COOH, -C1-6 alkylene- COOH, -N(Ci-6 alkyl)2 or                  OH       ; the capping group may be linked to the hydrophilic group via any linking group.

[0008] In a specific embodiment of the present invention, T1, T2 or T3 is selected from hydrophilic groups comprising monosaccharides, disaccharides, or oligosaccharides; or selected from hydrophilic groups comprising multiple (2 or more) carboxyl groups, multiple (2 or more) sulfonic acid groups, or chelating groups.

[0009] In a specific embodiment of the present invention, T1, T2 or T3 is selected from H, OH, methyl, carboxyl, -CH2CH2COOH, -N(CH2)2, OH OH OH OH OH In a specific embodiment of the present invention, the monovalent hydrophilic group Hp1 is preferably: -(CH2CH2O)n1-H, -(CH2CH2O)n1-CH3, , -(CH2)m2-X-(CH2)m3-COOH; preferably, the divalent hydrophilic group Hp2 is selected from: OH -(CH2CH2O)n1-,

[0010] In a specific embodiment of the present invention, the formula (Ia) or formula (Ib) has the following structures: M(Hp1)-Z1-Tr1-L1-D (Ia-1) M-Z1-Hp2-Z2-Tr1-L1-D (Ia-2) M-Z1(Hp1)-Z2-Tr1-L1-D (Ia-3) M(Hp1)-Z1-Hp2-Z2-Tr1-L1-D (Ia-4) M-Z1(Hp1)-Hp2-Z2-Tr1-L1-D (Ia-5) M(Hp1)-Z1(Hp1)-Hp2-Z2-Tr1-L1-D (Ia-6) M(Hp1)-Z1-L2(Tr2)-D (Ib-1) M-Z1-Hp2-Z2-L2(Tr2)-D (Ib-2) M-Z1(Hp1)-Z2-L2(Tr2)-D (Ib-3) M(Hp1)-Z1-Hp2-Z2-L2(Tr2)-D (Ib-4) M-Z1(Hp1)-Hp2-Z2-L2(Tr2)-D (Ib-5) M(Hp1)-Z1(Hp1)-Hp2-Z2-L2(Tr2)-D (Ib-6); wherein Z2 is a chemical bond or a divalent group, and Z1 is a divalent group or a trivalent group substituted with Hp1.

[0011] In a specific embodiment of the present invention, M is selected from the following structures: (Rb)r Lg is absent or is a leaving group, and the leaving group is selected from halogen, sulfonyl, trifluoromethanesulfonyl, and methanesulfonyl; ring B is selected from 5-14-membered heteroaromatic rings, or 3-14-membered heterocyclic rings; each Rb is the same or different and is independently selected from the following groups: halogen, cyano, oxo (=O), C1-6 alkyl, halo-C1-6 alkyl, hydroxy-C1-6 alkyl, C1-6 alkoxy or C3-8 cycloalkyl, 3-8-membered heterocyclyl, C1-6 alkyl-O-C1-6 alkyl-, C1-6 alkyl-(5-6-membered) heteroaryl-, or a monovalent hydrophilic group; r is an integer from 0 to 4; Lm1 is absent, or is selected from the following groups that are unsubstituted or optionally substituted with one, two or more Rm1: C6-14 aryl, 5-14-membered heteroaryl, 3-14-membered heterocyclyl; each Rm1 is the same or different and is independently selected from H, halogen, cyano, C1-6 alkyl or HOOC-C1-3 alkylene; Lm2 is selected from the following groups that are unsubstituted or optionally substituted with one, two or more Rm2: -(CH2)s-(C=O)-, or -C=C-(CH2)t-(C=O)-; each Rm2 is the same or different and is independently selected from H, halogen, cyano, C1-6 alkyl or -C1-6 alkylene-COOH, wherein the alkylene is optionally interrupted by one, two or more of the following groups: O, NH; s and t are the same or different and are independently an integer from 0 to 10; According to some embodiments, ring B is selected from a 5-6-membered N-containing heteroaromatic ring, or a 3-6 membered N-containing heterocyclic ring; According to some embodiments, ring B is selected from pyrimidine rings, pyridine rings, N J Oh triazine rings (such as N ),          ; According to some embodiments, each Rb is the same or different and is independently selected from cyano, oxo (=O), methoxy, cyclopropyl, trifluoromethyl, N \ , or -O-(CH2CH2O)n1-CH3; n1 is an integer from 1 to 36; According to some embodiments, Lm1 is absent or is selected from the following groups that are unsubstituted or optionally substituted with one, two or more Rm1: phenyl, piperidinyl or piperazinyl; According to some embodiments, Lm1 is selected from On7 \iO nO , or According to some embodiments, Lm2 is selected from the following groups that are unsubstituted or optionally substituted with one, two, or more Rm2: -CH2-(C=O)-, -(CH2)2-(C=O)-, -(CH2)5-(C=O)-, -C=C-(CH2)3-(C=O)-; According to some embodiments, formula M is formula M-1 as follows: (M-1) wherein t is an integer from 0 to 10; Z is N or CR22; R21, R22 and R23 are each independently selected from H, halogen, cyano, C1-6 alkyl, halo-C1-6 alkyl, hydroxy C1-6 alkyl, C1-6 alkoxy, C3-8 cycloalkyl, 3-8-membered heterocyclyl, C1-6 alkyl-O-C1-6 alkylene, or a monovalent hydrophilic group; provided that when Z is N, R21 and R23 are not both H; According to some embodiments, t is selected from 1, 2, 3, 4, 5, or 6; According to some embodiments, Z is N or C-CN; According to some embodiments, R21 is selected from H, halogen, cyano, C1-6 alkoxy, or C1-6 alkyl-O-C1-6 alkylene; According to some embodiments, Z is N, and R21 is selected from halogen, cyano, C1-3 alkoxy, or C1-3 alkyl-O-C1-3 alkylene; According to some embodiments, Z is N, and R21 is selected from methoxy or CH3-O-CH2-; According to some embodiments, Z is C-CN, and R21 is selected from H; According to some embodiments, R23 is selected from H.

[0012] According to some embodiments, M is selected from

[0013] In a specific embodiment of the present invention, Z1-Hp2-Z2 has the following structures: -Hp2-; -NH-(CH2)m6-HP2-; -NH-(CH2)m6-HP2-(CH2)m7CO-; -CO-(CH2)m6-HP2-; -CO-(CH2)m6-HP2-(CH2)m7CO-; -CO-(CH2)m6-HP2-(CH2)m7O(CH2)m8CO-; wherein m5 is an integer from 1 to 10.

[0014] Preferably, M-Z1-Hp2-Z2 has the following structures:

[0015] In a specific embodiment of the present invention, Z1(Hp1)-Z2 has the following structures: N(-Z6-Hp1)- -N(-Z6-Hp1)-(CH2)m9CO- -N(-Z6-Hp1)-(CH2)m9O(CH2)m10CO- O CH2 CH2 CH2 h2c-.._ 2    NH Z4---Hpi Hp^ f5 HN. .0 0 wherein Z3-Z7 are chemical bonds or divalent groups; preferably, Z3-Z6 are selected from C1-10 alkylene,-O-C1-10 alkylene-, -O-C1-10 alkylene-O-,-O-C1-10 alkylene-CO-, -O-C1-10 alkylene-NH-, -NH-C1-10 alkylene-, -NH-C1-10 alkylene-O-, -NH-C1-10 alkylene-CO-, -C1-10 alkylene-CO-, or -NH-C1-10 alkylene-NH-; m9 and m10 are each an integer from 1 to 10; preferably, M-Z1(Hp1)-Z2 has the following structures:

[0016] In a specific embodiment of the present invention, Z1(Hp1)-Hp2-Z2 has the following structures: -N(Hp1)-Hp2- -N(Hp1)-Hp2-(CH2)m11CO- -N(Hp1)-Hp2-(CH2)m11O(CH2)m12CO-; m11 and m12 are each an integer from 1 to10.

[0017] Preferably, M-Z1(Hp1)-Hp2-Z2- has the following structures:

[0018] According to some embodiments, Tr1 is glycine-glycine-phenylalanine-glycine (GGFG), GGFG or it is valine-alanine (VA), i.e.:

[0019] In a specific embodiment of the present invention, the structure of L2 is: D, wherein Xi is O or NH.

[0020] According to some embodiments, Tr2 is a valine-alanine (Val-Ala) or ^-glucuronide group, i.e. [002i] In a specific embodiment of the present invention, the structure of M-Zi-Hp2-Z2-L2(Tr2)- is: OH

[0022] In a specific embodiment of the present invention, the compound represented by formula (Ia) or formula (Ib) is preferably: wherein -Z1(Hp1)- is preferably: N(-Z6-Hp1)- -N(-Z6-Hp1)-(CH2)m9CO- -N(-Z6-Hp1)-(CH2)m9O(CH2)m10CO- CH2 CH2 H2C NH Z4---Hpi Hpi^ , 5 HN. .0 0 wherein Z3-Z7 are chemical bonds or divalent groups; preferably, Z3-Z6 are selected from C1-10 alkylene, -O-C1-10 alkylene-, -O-C1-10 alkylene-O-, -O-C1-10 alkylene-CO-, -O-C1-10 alkylene-NH-, -NH-C1-10 alkylene-, -NH-C1-10 alkylene-O-, -NH-C1-10 alkylene-CO-, -C1-10 alkylene-CO-, or -NH-C1-10 alkylene-NH-; t2, m9 and m10 are each independently an integer from 1 to 10; Hp3 is H or Hp1, and the remaining groups are as defined herein.

[0023] In a specific embodiment of the present invention, the compound represented by formula (Ia) or formula (Ib) is preferably: \ O           \ o                     o               o              o                    p \ / /                     \ / / \ / / \ / /                     \ / /                                  \ / / 0 = 0)              0=0)                             C / y                     0)x                 OK                           ok kz                      nK "°           "°      N^\ ° Ph yk              ’kkz     ’Kf i" HN. .0 0 H               / n6~~Tr1—Ld-D , OH < HN^O 0 Tr2 H            416^ L2—D , (CH2CH2O)n1.(CH2)q1 ^NZ                   / N'^kpTr1-L1-D 'X                   ° X(CH2CH2O)nr(CH2)q2 , (CH2CH2O)n1.(CH2)q1 \ /       \ Tr2 N                                 N         l-2 D \ 0 X(CH2CH2O)nr(CH2)q2 , ?!—L!—D 1 , tr2 LrD Tl IE wherein t2, n7, m13, and m14 are each independently an integer from 1 to 10; In a specific embodiment of the present invention, the compound represented by formula (Ia) or formula (Ib) is preferably: wherein Z, R21, R23, n1, n2, Tr1, L1, T1, T2, m13, m14, and D are as defined herein.

[0024] In a specific embodiment of the present invention, the compound represented by formula (Ia) or formula (Ib) is preferably: or wherein Z, R21, R23, and n1 are as defined herein.

[0025] preferably, n1 is an integer from 2 to 12, such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12; more preferably, n1 is an integer from 4 to 10.

[0026] In a specific embodiment of the present invention, the compound represented by formula (Ia) or formula (Ib) is preferably:

[0027] In a specific embodiment of the present invention, preferably, D is:

[0028] In a specific embodiment of the present invention, the compound represented by formula (Ia) or formula (Ib) is further preferably 001-056 or 001'-003': 48 ?Me           H      0 NY'’^YrNAAl H J    0 J  AY 0- N            °^NH      0 J HOOC^^ HOOC^n-^       036 <N^-x HOOC     / N<^COOH O fA u :  0 u     f 0H YAUvYJU ho^J °\A                        ° #T'OH                          037 O OH     OH     OH     0H ° 1 / 0 | o^ oY rN^ rN^ rNC A) NY^ ohnV° ohnV° oHN A Y° c H       H       H       H 038 oj S\ Ax z Y 31 NyY   g        ° V ' ; " 1 H ( 039 Y°Y h 3 h ?          rN^N^N^yN- O V J 0        0 o. A             0 / S N II 0                                         040 Ho3) / / ° XXNr€ / n ^vA A\ / N A h r jy o ? 0 ^~0H    H 1              /           n N             N i          N °<A    o    AAn i            \ A A / ° OH                 O N ,S' oz x 5 A o ho / .L^ AC0 ' Y H ° ArNY A ' n i fN A ^i Y H Y H LJ ? A o HO / ..JA Yo YnYa " £ H A J H         H        \ / A \z----xAA □ H 5 A H jA YhYnTYYna° Y1 'W / 0 HO"jA\ A o

[0029] The present invention further provides an antibody-drug conjugate represented by formula (IIa) or (IIb) obtained from formula (Ia) or formula (Ib), Ab-[M’-Zi-Tri-Li-D]p (IIa) or Ab-[M’-Zi-L2(Tr2)-D]p (IIb) wherein Ab is an antibody or an antigen-binding fragment thereof, M' is a fragment formed by conjugating M with Ab; P is selected from integers or decimals between 1 and 10. Zi, Tri, L1,D, Tr2, and L2 are as defined herein.

[0030] According to some embodiments, Ab is an antibody or an antigen-binding fragment, and the antigen-binding fragment is selected from Fab, Fab', (Fab’)2, Fd, Fv, disulfide-linked Fv, scFv, di-scFv, (scFv)2, a diabody, and a single domain antibody (sdAb); and / or the antibody is a murine antibody, a humanized antibody, a chimeric antibody, a bispecific antibody, or a multispecific antibody. [003i] According to some embodiments, Ab is an anti-HER2 antibody or an antigen-binding fragment thereof;

[0032] Preferably, the heavy chain variable region (VH) of the antibody or antigen-binding fragment thereof may include the following CDRs or variants of these CDRs: VHCDRi as set forth in SEQ ID NO. i: GFNIKDTYIH VHCDR2 as set forth in SEQ ID NO. 2: RIYPTNGYTRYADSVKG VHCDR3 as set forth in SEQ ID NO. 3: WGGDGFYAMDY The light chain variable region (VL) of the antibody or antigen-binding fragment thereof may include the following CDRs or variants of these CDRs: VLCDRi as set forth in SEQ ID NO. 4: RASQDVNTAVA VLCDR2 as set forth in SEQ ID NO. 5: SASFLYS VLCDR3 as set forth in SEQ ID NO. 6: QQHYTTPPT

[0033] For example, Ab is trastuzumab or an antigen-binding fragment thereof, and the combination of the heavy chain variable region and the light chain variable region are as set forth in SEQ ID NO. 7 and SEQ ID NO. 8: SEQ ID NO. 7: EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYT RYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGT LVTVSSA SEQ ID NO. 8: DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVP SRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRT

[0034] According to some embodiments, 0 is selected from integers or decimals between 4 and 9 (e.g., 7, 7.71, 7.84, 7.92, 7.94, 7.97, 7.98, 7.99, 8, 8.02, 8.06, or 8.14).

[0035] According to some specific embodiments, the antibody-drug conjugate represented by formula (IIa) of the present invention has the following structures: or wherein Ab, Z, R21, R23, and n1 are as defined herein; preferably, n1 is an integer from 2 to 12, such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12; more preferably, n1 is an integer from 4 to 10.

[0036] The present invention further provides the following intermediates: wherein Z, R21, R23, n1, n2, Tr1, m13, and m14 are as defined herein; Y1 is selected from halogen, hydroxyl, OSu, or C1-6 alkoxy.

[0037] Preferably, Y1, Y3, and Y4 are the same or different and are each independently selected from hydroxyl, methoxy, ethoxy, isopropoxy, tert-butoxy, or OSu; According to some embodiments, Z is N or C-CN; According to some embodiments, R21 is selected from H, halogen, cyano, C1-6 alkoxy, or C1-6 alkyl-O-C1-6 alkylene; According to some embodiments, Z is N, and R21 is selected from halogen, cyano, C1-3 alkoxy, or C1-3 alkyl-O-C1-3 alkylene; According to some embodiments, Z is N, and R21 is selected from methoxy or CH3-O-CH2-; According to some embodiments, Z is C-CN, and R21 is selected from H; According to some embodiments, R23 is selected from H.

[0038] The present invention further provides a linker of formula (IIIa) or (IIIb): M-Z1-Tr1-L1’ (IIIa) or M-Z1-L2’(Tr2) (IIIb) wherein L1’ and L2’ are reactive forms of L1 and L2; preferably: when L1 is a chemical bond, L1’ is a reactive form of Tr1, for example, Tr1 is a peptide fragment having a carboxyl group or an active ester at the C-terminus; when L1 is -NH-CH2-, then L1’ is O^ CH3 , where the wavy line indicates the linking site to a peptide residue; for another example, ^'N when Li is H 0 0-¾ , then L1’ is a carbonate active ester form of p-aminobenzyl alcohol, for example, residue. , where the wavy line indicates the linking site to a peptide

[0039] preferably, the structure of the linker is: wherein Z, R21, R23, and n1 are as defined herein; Y2 is selected from halogen, hydroxyl, OSu, or C1-6 alkoxy; preferably, Y2 is selected from hydroxyl, methoxy, ethoxy, isopropoxy, tert-butoxy, or OSu; According to some embodiments, Z is N or C-CN; According to some embodiments, R21 is selected from H, halogen, cyano, C1-6 alkoxy, or C1-6 alkyl-O-C1-6 alkylene; According to some embodiments, Z is N, and R21 is selected from halogen, cyano, C1-3 alkoxy, or C1-3 alkyl-O-C1-3 alkylene; According to some embodiments, Z is N, and R21 is selected from methoxy or CH3-O-CH2-; According to some embodiments, Z is C-CN, and R21 is selected from H; According to some embodiments, R23 is selected from H.

[0040] The present invention further provides a linker as shown below, which is used for obtaining an antibody-drug conjugate formed by linking an antibody to a drug via the linker, wherein Z, R21, R23, n1, n2, Tr1, L1, T1, T2, m13, and m14 are as defined herein; position 1 is linked to Ab, and position 2 is linked to D.

[0041] According to some embodiments, the linker is as shown below: 67 wherein position 1 is linked to Ab and position 2 is linked to D.

[0042] The present invention further provides a pharmaceutical composition, which comprises a therapeutically effective amount of the antibody-drug conjugate represented by formula (IIa) or (IIb).

[0043] According to an embodiment of the present invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

[0044] According to an embodiment of the present invention, the pharmaceutical composition may further comprise one or more additional therapeutic agents.

[0045] The present invention further provides a method for treating a tumor disease, which comprises administering to a patient a prophylactically or therapeutically effective amount of at least one of the antibody-drug conjugate represented by formula (IIa) or (IIb), a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof.

[0046] The present invention further provides a method for treating a tumor disease, which comprises administering to a patient a prophylactically or therapeutically effective amount of the above pharmaceutical composition.

[0047] The tumor disease is selected from breast cancer, gastric cancer, lung cancer, colorectal cancer, large intestine cancer, ovarian cancer, liver cancer, kidney cancer, esophageal cancer, cervical cancer, bladder cancer, pancreatic cancer, prostate cancer, nasopharyngeal carcinoma, melanoma, or leukemia.

[0048] In some embodiment, the patient includes a mammal, preferably a human.

[0049] The present invention further provides use of at least one of the antibody-drug conjugate represented by formula (IIa) or (IIb), a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof, or a pharmaceutical composition thereof in the treatment of a tumor disease.

[0050] The present invention further provides use of at least one of the antibody-drug conjugate represented by formula (IIa) or (IIb), a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof, or the above pharmaceutical composition in the manufacture of a topoisomerase I inhibitor and / or in the manufacture of a drug for preventing or treating a disease or condition associated with topoisomerase I.

[0051] In some embodiments, the disease or condition is tumor, comprising breast cancer, gastric cancer, lung cancer, colorectal cancer, large intestine cancer, ovarian cancer, liver cancer, kidney cancer, esophageal cancer, cervical cancer, bladder cancer, pancreatic cancer, prostate cancer, nasopharyngeal carcinoma, melanoma, or leukemia. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Fig. 1 Schematic diagram of free toxin release of ADC-2 in mouse, rat, monkey, and human plasma; Fig. 2 Schematic diagram of free toxin release of ADC-12 in mouse, rat, monkey, and human plasma; Fig. 3 Schematic diagram of free toxin release of DS8201 in mouse, rat, monkey, and human plasma; Fig. 4 Schematic diagram of efficacy evaluation in NCI-N87 tumor-bearing mice; Fig. 5 Schematic diagram of efficacy evaluation in JIMT-1 tumor-bearing mice; Fig. 6 Schematic diagram of efficacy evaluation in RT11284 tumor-bearing mice; Fig. 7 Schematic diagram of efficacy evaluation in HCT116 tumor-bearing mice; Fig. 8 Schematic diagram of efficacy evaluation in Capan-1 tumor-bearing mice; Fig. 9 Schematic diagram of efficacy evaluation in NCI-N87-Enhertu-resistant cell tumorbearing mice; Fig. 10 Schematic diagram of efficacy evaluation in human gastric cancer LD1-0017-411335 PDX tumor-bearing mice; Fig. 11 PK profile of ADC-2 in mice; Fig. 12 Comparison of in vivo tumor inhibitory effects of ADC-2 and ADC-58 in JIMT-1 xenograft model; Fig. 13 Comparison of in vivo tumor inhibitory effects of ADC-2 and ADC-58 in NCI-N87 xenograft model; Fig. 14. Results of ADC-58 plasma stability test; Fig. 15 Comparison of in vivo tumor inhibitory effects of ADC-2 and ADC-57 in 22RV1 xenograft model. Definition and Explanation of Terms

[0053] Unless otherwise indicated, the definitions of groups and terms recorded in the specification and claims of the present application, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, definitions of specific compounds in examples, etc., may be arbitrarily combined and associated with each other. Such combined and associated group definitions and compound structures should be understood to be within the scope of the specification and / or claims of the present application.

[0054] The "linker", "linker structure" or "connector" or "linking unit" mentioned in the present invention refers to a chemical structure fragment or bond that is connected to an antibody at one end and to a drug (pharmaceutical compound) at the other end, and can also be connected to other linkers before being connected to the pharmaceutical compound. The linker structure of the present invention can be synthesized by methods known in the art, or by the method described in the present invention.

[0055] As used herein, "polyethylene glycol" refers to a homogeneous compound containing 236 ethoxy units. In the present invention, "polyethylene glycol" includes homogeneous polyethers formed by direct linkage of ethoxy units, as well as homogeneous compounds obtained by chemical reactions of compounds containing ethoxy units (e.g., NH2-CH2CH2O-CH2CH2O-CH2-COOH). That is, any number of ethoxy units in the "polyethylene glycol" may be spaced or interrupted by optional chemical groups such as amide groups. Monovalent polyethylene glycol groups can be derived from homogeneous monofunctional polyethylene glycol derivatives (e.g., a-methoxy-w-carboxy oligoethylene glycol), and divalent polyethylene glycol groups can be derived from homogeneous bifunctional polyethylene glycol derivatives (e.g., a-amino-w-carboxy oligoethylene glycol).

[0056] The "divalent group" or "divalent linking group" mentioned in the present invention refers to any divalent organic group, preferably selected from: -O-, -NH-, -CO-, -O-C1-6 alkylene-, -N(R0)-C1-6 alkylene-, -C1-6 alkylene-O-, -C1-6 alkylene-N(R0)-, -CO-C1-6 alkylene-, -C1-6 alkylene-CO-, -O-C1-6 alkylene-CO-, -C1-6 alkylene-CO-N(R0)-, -C1-6 alkylene-N(R0)-CO-, -CO-C1-6 alkylene-N(R0)-CO-C1-6 alkylene-, -CO-C1-6 alkylene-CO-N(R0)-C1-6 alkylene-, -C1-6 alkylene-CO-N(R0)-C1-6 alkylene-, -C1-6 alkylene-N(R0)-CO-C1-6 alkylene-, -C1-6 alkylene-N(R0)-CO-C1-6 alkylene-COO-C1-6 alkylene-, -C1-6 alkylene-CO-N(R0)-C1-6 alkylene-COO-C1-6 alkylene-, -CO-C1-6 alkylene-N(R0)-CO-C1-6 alkylene-COO-C1-6 alkylene-, -CO-C1-6 alkylene-CO-N(R0)-C1-6 alkylene-COO-C1-6 alkylene-, etc.; R0 is selected from C1-4 alkyl, such as methyl; preferably, the alkyl is optionally interrupted by an O atom.

[0057] The "trivalent group" mentioned in the present invention refers to any trivalent organic group, preferably obtained by substitution of the aforementioned "divalent group", or preferably a trivalent group based on an amino acid, a benzene ring, or a heterocyclic ring.

[0058] The "linking group" mentioned in the present invention includes the aforementioned "divalent group" or "trivalent group". It functions as a divalent group when connecting two functional fragments (e.g., the linkage between a capping group and a hydrophilic fragment, or between an antibody linker and a peptide fragment), and functions as a trivalent group when connecting three functional fragments (e.g., linking group L2 connecting a hydrophilic fragment, a trigger group, and a drug).

[0059] The "trigger group" mentioned in the present invention refers to a group that triggers drug release in a cleavable ADC linker. Examples include peptide fragments cleavable by cathepsins, or fragments cleavable by P-glucuronidase and P-galactosidase. The trigger group is often conjugated to the drug via a self-immolative group, releasing the drug through a cascade reaction under specific enzymatic or other conditions.

[0060] The "antibody-drug conjugate" (ADC) mentioned in the present invention refers to a targeting moiety connected to a biologically active drug via a stable linking unit.

[0061] The "bioactive molecules" mentioned in the present invention refer to cytotoxic drugs, which are chemical molecules that can strongly destroy the normal growth of tumor cells.

[0062] Unless otherwise indicated, the numerical ranges recited in the present specification and claims are equivalent to at least reciting each specific integer value therein. For example, a numerical range of "1 to 36" shall be deemed to specifically disclose each integer value in the numerical range "1 to 36", namely 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, and 36. In addition, when a numerical range is defined as a "number," it should be understood that both endpoints of the range, each integer within the range, and each decimal within the range are recited.

[0063] The term "an integer from 0 to 10" denotes 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.

[0064] The term "halogen" represents fluorine, chlorine, bromine, and iodine.

[0065] The "C1-10 alkyl" represents a linear and branched alkyl having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, the "C1-8 alkyl" represents a linear and branched alkyl having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, and the "C1-6 alkyl" represents a linear and branched alkyl having 1, 2, 3, 4, 5, or 6 carbon atoms. The alkyl is, e.g., methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, or 1,2-dimethylbutyl, etc., or isomers thereof.

[0066] The "C2-10 alkenyl" should be construed to represent preferably a linear or branched monovalent hydrocarbon radical comprising one or more double bonds and having 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, and is more preferably "C2-8 alkenyl". The "C2-10 alkenyl" should be construed to represent preferably a linear or branched monovalent hydrocarbon radical comprising one or more double bonds and having 2, 3, 4, 5, 6, 7, or 8 carbon atoms, for example, having 2, 3, 4, 5, or 6 carbon atoms (namely, C2-6 alkenyl) or having 2 or 3 carbon atoms (namely, C2-3 alkenyl). It should be understood that in the case where the alkenyl comprises more than one double bond, the double bonds may be separated from each other or conjugated. The alkenyl is, e.g., vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, pent-4-enyl, (E)-pent-3-enyl, (Z)-pent-3-enyl, (E)-pent-2-enyl, (Z)-pent-2-enyl, (E)-pent-1-enyl, (Z)-pent-1-enyl, hex-5-enyl, (E)-hex-4-enyl, (Z)-hex-4-enyl, (E)-hex-3-enyl, (Z)-hex-3-enyl, (E)-hex-2-enyl, (Z)-hex-2-enyl, (E)-hex-1-enyl, (Z)-hex-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl, (E)-1-methylprop-1-enyl, (Z)-1-methylprop-1-enyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, 1- methylbut-3-enyl, 3-methylbut-2-enyl, (E)-2-methylbut-2-enyl, (Z)-2-methylbut-2-enyl, (E)-1-methylbut-2-enyl, (Z)-1-methylbut-2-enyl, (E)-3-methylbut-1-enyl, (Z)-3-methylbut-1-enyl, (E)-2-methylbut-1-enyl, (Z)-2-methylbut-1-enyl, (E)-1-methylbut-1-enyl, (Z)-1-methylbut-1-enyl, 1,1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propyvinyl, or 1-isopropylvinyl.

[0067] The "C2-10 alkynyl" should be construed to represent preferably a linear or branched monovalent hydrocarbon radical comprising one or more triple bonds and having 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, for example, having 2, 3, 4, 5, 6, 7, or 8 carbon atoms (namely, "C2-8 alkynyl"), having 2, 3, 4, 5, or 6 carbon atoms (namely, "C2-6 alkynyl"), or having 2 or 3 carbon atoms (namely, "C2-3 alkynyl"). The alkynyl is, e.g., ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl, pent-1-ynyl, pent-2-ynyl, pent-3-ynyl, pent-4-ynyl, hex-1-ynyl, hex-2-ynyl, hex-3-ynyl, hex-4-ynyl, hex-5-ynyl, 1-methylprop-2-ynyl, 2-methylbut-3-ynyl, 1-methylbut-3-ynyl, 1-methylbut-2-ynyl, 3-methylbut-1-ynyl, 1-ethylprop-2-ynyl, 3-methylpent-4-ynyl, 2-methylpent-4-ynyl, 1-methylpent-4-ynyl, 2-methylpent-3-ynyl, 1-methylpent-3-ynyl, 4-methylpent-2-ynyl, 1-methylpent-2-ynyl, 4-methylpent-1-ynyl, 3-methylpent-1-ynyl, 2-ethylbut-3-ynyl, 1-ethylbut-3-ynyl, 1-ethylbut-2-ynyl, 1-propylprop-2-ynyl, 1-isopropylprop-2-ynyl, 2,2-dimethylbut-3-ynyl, 1,1-dimethylbut-3-ynyl, 1,1-dimethylbut-2-ynyl, or 3,3-dimethylbut-1-ynyl. In particular, the alkynyl is ethynyl, prop-1-ynyl, or prop-2-ynyl.

[0068] The term "C3-10 cycloalkyl" should be construed to represent saturated monovalent monocyclic, bicyclic (e.g., bridged cyclic or spirocyclic) hydrocarbon ring or tricyclic alkane having 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. The C3-10 cycloalkyl may be a monocyclic hydrocarbon radical such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or cyclodecyl, or may be a bicyclic hydrocarbon radical such as bornyl, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonyl, or 2,6-diazaspiro[3,4]octyl, or may be a tricyclic hydrocarbon radical such as adamantyl.

[0069] Unless otherwise defined, the term "3-6-membered heterocyclyl" refers to a saturated or unsaturated non-aromatic ring or ring system, for example, it is a 4-, 5-, or 6-membered monocyclic ring, and contains at least one, for example, 1, 2, 3, 4, 5, or more heteroatoms selected from O, S, and N, wherein N and S may further be optionally oxidized to various oxidation states to form nitrogen oxides, -S(O)-, or -S(O)2- states. The heterocyclyl may comprise a fused or bridged ring and a spirocyclic ring. In particular, the heterocyclyl may include, but is not limited to: a 4-membered ring, such as azetidinyl or oxetanyl; a 5-membered ring, such as tetrahydrofuryl, dioxolyl, pyrrolidyl, imidazolidinyl, pyrazolidyl, or pyrrolinyl; or a 6-membered ring, such as tetrahydropyranyl, piperidyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, or trithianyl.

[0070] The term "C6-14 aryl" should be construed to represent preferably a monovalent aromatic or partially aromatic monocyclic, bicyclic, or tricyclic hydrocarbon ring having 6, 7, 8, 9, 10, 11, 12, 13, or 14 carbon atoms ("C6-14 aryl"), particularly a ring having 6 carbon atoms ("C6 aryl"), such as phenyl; or biphenyl, or a ring having 9 carbon atoms ("C9 aryl"), such as indanyl or indenyl, or a ring having 10 carbon atoms ("C10 aryl"), such as tetralyl, dihydronaphthyl, or naphthyl, or a ring having 13 carbon atoms ("C13 aryl"), such as fluorenyl, or a ring having 14 carbon atoms ("C14 aryl"), such as anthranyl. When the C6-20 aryl is substituted, it can be monosubstituted or polysubstituted. Further, the substitution site is not limited, for example, may be ortho-, para-, or meta-substitution.

[0071] The term "5-14-membered heteroaryl" should be construed to comprise such a monovalent monocyclic, bicyclic, or tricyclic aromatic ring system that has 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 ring atoms, particularly 5 or 6 or 9 or 10 carbon atoms, and comprises from 1 to 5, preferably from 1 to 3, heteroatoms each independently selected from N, O, and S, and, further, may be additionally benzo-fused in each case. "Heteroaryl" also means a group in which a heteroaromatic ring is fused with one or more aryl, alicyclic, or heterocyclic rings, wherein the radical or site of attachment is on the heteroaromatic ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7- or 8-indolizinyl, 1-, 3-, 4-, 5-, 6-, or 7-isoindolyl, 2-, 3-, 4-, 5-, 6- or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-indazolyl, 2-, 4-, 5-, 6-, 7- or 8-purinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-quinolizinyl, 2-, 3- , 4-, 5-, 6-, 7- or 8-quinolinyl, 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolinyl, 1-, 4-, 5-, 6-, 7- or 8-phthalazinyl, 2-, 3-, 4-, 5- or 6-naphthyridinyl, 2-, 3-, 5-, 6-, 7- or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7- or 8-cinnolinyl, 2-, 4-, 6- or 7-pteridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-4aH carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-carbazolylcarbazolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8- or 9-carbolinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenanthridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8- or 9-acridinyl, 1-, 2-, 4-, 5-, 6-, 7-, 8- or 9-pyridyl, 2-, 3-, 4-, 5-, 6-, 8-, 9- or 10-phenanthrolinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-phenazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenothiazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenazinyl , 2-, 3-, 4-, 5-, 6- or 1-, 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-benzoisoquinolyl, 2-, 3-, 4- or thieno[2,3-b]furanyl, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-7H-pyrazino[2,3-c]carbazolyl, 2-, 3-, 5-, 6- or 7-2H-furo[3,2-b]pyranyl , 2-, 3-, 4-, 5-, 7-, or 8-5H-pyrido[2,3-d]-o-oxazinyl, 1-, 3-, or 5-1H-pyrazolo[4,3-d]-oxazolyl, 2-, 4-, or 54H-imidazo[4,5-d]thiazolyl, 3-, 5-, or 8-pyrazino[2,3-d]pyridazinyl, 2-, 3-, 5-, or 6-imidazo[2,1-b]thiazolyl, 1-, 3-, 6-, 7-, 8-, or 9 -furo[3,4-c]cinnolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10 or 11-4H-pyrido[2,3-c]carbazolyl, 2-, 3-, 6- or 7-imidazo[1,2-b][1,2,4]triazinyl, 7-benzo[b]thienyl, 2-, 4-, 5-, 6- or 7-benzoxazolyl, 2-, 4-, 5-, 6- or 7-benzimidazolyl, 2-, 4- , 4-, 5-, 6- or 7-benzothiazolyl, 1-, 2-, 4-, 5-, 6-, 7-, 8- or 9-benzoxapinyl, 2-, 4-, 5-, 6-, 7- or 8-benzoxazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-4H-pyrrolo[1,2-b][2]benzazapinyl. A typical fused heteroaryl includes, but is not limited to, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, and 2-, 4-, 5-, 6-, or 7-benzothiazolyl. The 5-20-membered heteroaryl may be linked to other groups to form the compound of the present invention by linking a carbon atom on a 5-20-membered heteroaryl ring to the other groups, or by linking a heteroatom on a 520-membered heteroaryl ring to the other groups. When the 5-20-membered heteroaryl is substituted, it may be monosubstituted or polysubstituted. Further, the substitution site is not limited, for example, hydrogen attached to a carbon atom on the heteroaryl ring may be substituted, or hydrogen attached to a heteroatom on the heteroaryl ring may be substituted.

[0072] The term "spirocyclic ring" refers to a ring system in which two rings share 1 ringforming atom.

[0073] The term "fused ring" refers to a ring system in which two rings share 2 ring atoms.

[0074] The term "bridged ring" refers to a ring system in which two rings share more than 3 ring-forming atoms.

[0075] Wavy lines (  ) intersecting with chemical bonds are used to indicate positions where a group is linked to other atoms or groups in the general structure.

[0076] Unless otherwise specified, the heterocyclyl, heteroaryl, or heteroarylene comprises all possible isomeric forms thereof, such as positional isomers thereof. Therefore, for some illustrative non-limiting examples, the substitution or bonding to other radicals may be comprised at, e.g., 1, 2, or more of its 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-C, (if present), comprising pyridin-2-yl, pyridyliden-2-yl, pyridin-3-yl, pyridyliden-3-yl, pyridin-4-yl, and pyridyliden-4-yl; and the thienyl or thienylenyl comprises thien-2-yl, thienylen-2-yl, thien-3-yl, and thienylen-3-yl; pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, and pyrazol-5-yl.

[0077] The term "alkoxy" refers to -O-(alkyl), wherein the alkyl is as defined above. Nonlimiting examples of alkoxy comprise: methoxy, ethoxy, propoxy, and butoxy. The alkoxy may be optionally substituted or unsubstituted, and may be, when substituted, substituted with a substituent which is preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylamino, halogen, sulfydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, or heterocycloalkoxy.

[0078] The term "alkylamino" refers to -NH-(alkyl), wherein the alkyl is as defined above. Nonlimiting examples of alkylamino comprise, for example, methylamino, ethylamino, propylamino, isopropylamino, and butylamino.

[0079] The term "(alkyl)2amino" refers to -N-(alkyl)2, wherein the alkyl is as defined above. Non-limiting examples of (alkyl)2 amino comprise: for example, dimethylamino, methylethylamino, diethylamino, dipropylamino, methylpropylamino, diisopropylamino, and dibutylamino.

[0080] The "haloalkyl" refers to alkyl substituted with one or more halogens, wherein the alkyl is as defined above.

[0081] The term "antibody" refers to an immunoglobulin-derived molecule capable of specifically binding to a target antigen, and the immunoglobulin-derived molecule binds to the target antigen through at least one antigen-binding site located in a variable region thereof. When referring to the term "antibody", unless otherwise specified in the context, it includes not only an intact antibody but also an antigen-binding fragment capable of specifically binding to a target antigen. An "intact antibody" typically consists of two pairs of polypeptide chains, each pair having one light chain (LC) and one heavy chain (HC). Light chains of the antibody can be classified as kappa (k) and lambda (X) light chains. Heavy chains can be classified as p, 5, y, a or a, and the isotype of the antibody is respectively defined as IgM, IgD, IgG, IgA, and IgE. Within light and heavy chains, the variable and constant regions are joined by a "J" region of about 12 or more amino acids, and the heavy chain also comprises a "D" region of about 3 or more amino acids. Each heavy chain consists of a heavy chain variable region (VH) and a heavy chain constant region (CH). The heavy chain constant region consists of 3 domains (CH1, CH2, and CH3). Each light chain consists of a light chain variable region (VL) and a light chain constant region (CL). The light chain constant region consists of one domain CL. The constant domain is not directly involved in binding of an antibody to an antigen, but exhibits various effector functions, such as mediating the binding of immunoglobulins to host tissues or factors, including binding of various cells (e.g., effector cells) of the immune system and the first component (C1q) of the classical complement system. VH and VL regions can be further subdivided into regions with high variability (called complementarity determining regions (CDRs)) interspersed with relatively conserved regions called framework regions (FRs). Each of the VH and VL consists of 3 CDRs and 4 FRs arranged from an amino terminal to a carboxyl terminal in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions (VH and VL) of each heavy chain / light chain pair respectively form an antigen-binding site. The distribution of amino acids in each region or domain can be based on the definitions in Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987 and 1991)), or Chothia & Lesk (1987) J. Mol. Biol. 196:901-917; Chothia et al. (1989) Nature 342:878-883. As used herein, the term "complementarity determining region" or "CDR" refers to amino acid residues responsible for antigen binding in a variable region of an antibody. Each of the variable regions of the heavy and light chains comprises three CDRs, named CDR1, CDR2, and CDR3. The precise boundaries of these CDRs can be defined according to various numbering systems known in the art, such as the definitions in the Kabat numbering system (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991), the Chothia numbering system (Chothia & Lesk (1987) J. Mol. Biol. 196:901-917; Chothia et al. (1989) Nature 342:878-883), or the IMGT numbering system (Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003). For a given antibody, a person of skill in the art will readily identify the CDRs defined by each numbering system. Furthermore, the corresponding relationship between different numbering systems is well known to a person of skill in the art (for example, see Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003). In the present invention, the CDR comprised in the antibody or the antigen-binding fragment thereof of the present invention can be determined according to various numbering systems known in the art. In some embodiments, the CDR comprised in the antibody or antigen-binding fragment thereof of the present invention is preferably determined by the Kabat numbering system. As used herein, the term "framework region" or "FR" residues refer to those amino acid residues in variable regions of an antibody other than the CDR residues as defined above. The term "antibody" is not limited to any particular method of producing the antibody. For example, antibodies include recombinant antibodies, monoclonal antibodies, and polyclonal antibodies. Antibodies can be of different isotypes, for example, IgG (e.g., IgG1, IgG2, IgG3, or IgG4 subtype), IgA1, IgA2, IgD, IgE, or IgM antibodies. As used herein, the term "antigen-binding fragment" of an antibody refers to a polypeptide comprising a fragment of a full-length antibody that retains the ability to specifically bind to the same antigen that the fulllength antibody binds to and / or competes with the full-length antibody for specific binding to the antigen, which is also referred to as an "antigen-binding portion". Generally, see Fundamental Immunology, Ch. 7 (Paul, W., ed., 2nd ed., Raven Press, N.Y. (1989), which is incorporated herein by reference in its entirety for all purposes. The antigen-binding fragment of an antibody can be produced by recombinant DNA technologies or by enzymatic or chemical cleavage of an intact antibody. Non-limiting examples of the antigen-binding fragment include Fab, Fab', F(ab')2, Fd, Fv, disulfide-linked Fv, scFv, di-scFv, a diabody, a single-domain antibody, and a polypeptide comprising at least a portion of an antibody sufficient to confer a specific antigenbinding ability to the polypeptide. Engineered antibody variants are reviewed in Holliger et al., 2005; Nat Biotechnol, 23: 1126-1136. As used herein, the term "Fd" means an antibody fragment consisting of VH and CH1 domains; the term "Fab fragment" means an antibody fragment consisting of VL, VH, CL, and CH1 domains; the term "F(ab')2 fragment" means an antibody fragment comprising two Fab fragments linked by a disulfide bridge on a hinge region; and the term "Fab' fragment" means a fragment obtained after reducing disulfide bonds linking two heavy chain fragments in the F(ab')2 fragment, consisting of a complete light chain and the Fd fragment (consisting of VH and CH1 domains) of a heavy chain. As used herein, the term "Fv" means an antibody fragment consisting of the VL and VH domains of a single arm of an antibody. The Fv fragment is generally considered to be the smallest antibody fragment that capable of forming a complete antigen-binding site. It is generally believed that six CDRs confer an antigen-binding specificity to an antibody. However, a single variable region (e.g., the Fd fragment, which comprises only three CDRs specific for an antigen) also has the ability to recognize and bind to an antigen, although the affinity thereof may be lower than that of a complete binding site. As used herein, the term "Fc" means an antibody fragment formed by the second and third constant regions of a first heavy chain of an antibody binding to the second and third constant regions of a second heavy chain of the antibody through disulfide bonds. The Fc fragment of an antibody has a variety of different functions but is not involved in antigen binding. The "effector functions" mediated by the Fc domain comprise Fc receptor binding; Clq binding and complement-dependent cytotoxicity (CDC); antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g., B cell receptor); and B cell activation, or the like. The Fc domain may comprise both a native Fc region and a variant Fc region. A native Fc region comprises an amino acid sequence that is consistent with the amino acid sequence of an Fc region found in nature, for example, a native sequence human Fc region comprises a native sequence human IgG1 Fc region; a native sequence human IgG2 Fc region; a native sequence human IgG3 Fc region; and a native sequence human IgG4 Fc region, as well as naturally occurring variants thereof. A variant Fc region comprises an amino acid sequence which differs from the amino acid sequence of a native sequence Fc region by at least one amino acid modification. In some embodiments, a variant Fc region may possess altered effector functions (e.g., Fc receptor binding, antibody glycosylation, number of cysteine residues, effector cell function, or complement function) compared to a native Fc region. As used herein, the term "scFv" refers to a single polypeptide chain comprising VL and VH domains, wherein the VL and VH are linked by a linker. Such scFv molecules may have a general structure: NH2-VL-Linker-VH-COOH or NH2-VH-Linker-VL-COOH. A suitable linker in the prior art consists of repeated GGGGS amino acid sequences or variants thereof. For example, a linker having an amino acid sequence (GGGGS)4 may be used, but a variant thereof may also be used. In some cases, a disulfide bond may also exist between the VH and VL of the scFv. In some embodiments of the present invention, the scFv may form di-scFv, which refers to two or more single scFvs in series to form an antibody. In some embodiments of the present invention, the scFv may form (scFv)2, which refers to two or more single scFvs in parallel to form an antibody. As used herein, the term "diabody" means that the VH and VL domains thereof are expressed on a single polypeptide chain, but it uses a linker that is too short to allow pairing between two domains of the same chain, thereby forcing the domains to pair with the complementary domains of another chain and forming two antigen-binding sites (see, e.g. Holliger P. et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993), and Poljak R.J. et al., Structure 2:1121-1123 (1994)). As used herein, the term "single-domain antibody (sdAb)" has the meaning commonly understood by a person of skill in the art, and refers to an antibody fragment consisting of a single monomeric variable antibody domain (e.g., single heavy chain variable region), which retains the ability to specifically bind to the same antigen as the fulllength antibody. As used herein, the term "bispecific antibody" refers to an antibody having a binding specificity for two different antigens (or epitopes). The term "multispecific antibody" refers to an antibody having a binding specificity for at least more than two (e.g., three or four) different antigens (or epitopes). The bispecific antibody or the multispecific antibody comprises multiple antigen-binding domains having a binding specificity for different antigens (or epitopes), and thus is able to bind to at least two different binding sites and / or target molecules. Each antigen-binding domain comprised in the bispecific antibody or the multispecific antibody can be independently selected from a full-length antibody (e.g. IgG antibody) or an antigenbinding fragment thereof (e.g. Fv fragment, Fab fragment, F(ab')2 fragment, or scFv). In some cases, individual antigen-binding domains are linked by peptide linkers. Each of the above antibody fragments retains the ability to specifically bind to the same antigen as the full-length antibody, and / or competes with the full-length antibody for specific binding to the antigen. An antigen-binding fragment of an antibody (e.g., the above-mentioned antibody fragment) can be obtained from a given antibody (e.g., antibody provided in the present invention) by a conventional technology known to those skilled in the art (e.g., recombinant DNA technology or enzymatic or chemical cleavage method), and the antigen-binding fragment of the antibody can be screened for specificity in the same manner as for an intact antibody. As used herein, the term "humanized antibody" refers to a non-human antibody that is genetically engineered, and the amino acid sequence thereof is modified to increase the homology with the sequence of a human antibody. Generally speaking, all or part of CDR regions of a humanized antibody are derived from a non-human antibody (donor antibody), and all or part of non-CDR regions (e.g., variable region FR and / or constant region) are derived from a human immunoglobulin (receptor antibody). In some embodiments, the CDR regions of a humanized antibody are derived from a non-human antibody (donor antibody), and all or part of non-CDR regions (e.g., variable region FR and / or constant region) are derived from a human immunoglobulin (receptor antibody). A humanized antibody generally retains the desired properties of a donor antibody, including but not limited to, antigen specificity, affinity, reactivity, etc. In the present application, the donor antibody may be a murine antibody having desired properties (e.g., antigen specificity, affinity, reactivity). To prepare a humanized antibody, CDR regions of a donor antibody can be inserted into human framework sequences by a method known in the art. In some cases, the human framework sequence may comprise amino acid mutations that are substituted with corresponding non-human residues. In addition, a humanized antibody may also comprise residues that are not found in a variable region (e.g., light chain variable region or heavy chain variable region) of an initial donor antibody or in a human framework sequence to further improve or optimize the performance of the humanized antibody. As used herein, the term "chimeric antibody" refers to an antibody in which a portion of the light chain and / or the heavy chain thereof is derived from one antibody (which may be derived from a particular species or belong to a particular antibody type or subtype), and another portion of the light chain and / or the heavy chain is derived from another antibody (which may be derived from the same or different species or belong to the same or different antibody types or subtypes), but in any case, the binding activity to a target antigen is still retained. In some embodiments, the term "chimeric antibody" may include an antibody in which the heavy chain variable region and light chain variable region of the antibody are derived from a first antibody, and the heavy chain constant region and light chain constant region of the antibody are derived from a second antibody.

[0082] In the present invention, the compound involved also comprises an isotopically labeled compound, which is the same as the compound of formula I, but in which one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number usually occurring in nature. Exemplary isotopes that can be incorporated into the compound of the present invention comprise isotopes of H, C, N, O, S, F and Cl, such as 2H, 3H, 13C, 11C, 14C, 15N, 18O, 17O, 32P, 35S, 18F and 36Cl. The compound of the present invention containing the aforementioned isotopes and / or other isotopes of other atoms and a prodrug thereof, or a pharmaceutically acceptable salt of the compound or the prodrug are covered in the scope of the present invention. Some isotopically labeled compounds of the present invention, for example, compounds incorporated with radioactive isotopes (such as 3H and 14C) can be used in the distribution determination of drugs and / or substrates in tissues. The isotopes tritium (that is, 3H) and carbon 14 (that is, 14C) are particularly preferred because of their easy preparation and detectability. Furthermore, the substitution with a heavier isotope (such as deuterium, that is, 2H or D) can provide some therapeutic advantages due to the greater metabolic stability (for example, increased in-vivo half-life or reduced dosage requirement), and is thus preferred in some cases. The compound of the present invention as claimed in the claims can be particularly defined to be substituted with deuterium or tritium. In addition, in case that hydrogen in a substituent is not explicitly indicated to be deuterium or tritium, it does not mean that deuterium or tritium is excluded, but deuterium or tritium can be contained as well.

[0083] Those skilled in the art will appreciate that the compound represented by formula (I) may exist in the form of various pharmaceutically acceptable salts. If the compounds have basic centers, they can form acid addition salts; if they have acidic centers, they can form base addition salts; if the compounds contain both acidic centers (e.g. carboxyl) and basic centers (e.g. amino), they can also form inner salts.

[0084] The compounds of the present invention may be present in the form of solvates, such as hydrates, wherein the compounds of the present invention comprise polar solvents, such as water, methanol or ethanol, as structural elements of the lattice of the compounds. The polar solvent, especially water, may be present in a stoichiometric or non-stoichiometric amount.

[0085] Depending on their molecular structure, the compounds of the present invention may be chiral and may therefore exist in various enantiomeric forms. These compounds may therefore exist in racemic or optically active forms. The compounds of the present invention encompass isomers in which the chiral carbons are each in R or S configuration, or mixtures and racemates thereof. The compounds of the present invention or their intermediates can be separated into enantiomeric compounds by chemical or physical methods known to those skilled in the art, or used in synthesis in this form. In the case of racemic amines, diastereomers are prepared from the mixture by reaction with an optically active resolving reagent. Examples of suitable resolving reagents are optically active acids, such as the R and S forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, appropriate N-protected amino acids (e.g. N-benzoylproline or N-phenylsulfonylproline) or various optically active camphorsulfonic acids. Chromatographic enantiomeric resolution can also be advantageously carried out by means of optically active resolving reagents, such as dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chirally derivatized methacrylate polymers immobilized on silica gel. Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, for example, hexane / isopropanol / acetonitrile.

[0086] The corresponding stable isomers can be separated according to known methods, such as by extraction, filtration or column chromatography.

[0087] The term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, and most preferably humans.

[0088] The term "therapeutically effective amount" refers to the amount of an active compound or drug which causes a biological or medical response sought by a researcher, veterinarian, physician or other clinician in a tissue, system, animal, individual or human, and which comprises one or more of the following: (1) prevention of disease: for example, prevention of a disease, disorder or condition in an individual who is susceptible to the disease, disorder or condition but has not yet experienced or developed the pathology or symptoms of the disease. (2) Inhibition of disease: for example, inhibition of a disease, disorder or condition (i.e., prevention of further progression of pathology and / or symptoms) in an individual undergoing or exhibiting the pathology or symptoms of the disease, disorder or condition. (3) Alleviation of disease: for example, alleviation of a disease, disorder or condition (i.e., reversion of pathology and / or symptoms) in an individual undergoing or exhibiting the pathology or symptoms of the disease, disorder or condition. Abbreviation List PBS Phosphate buffered saline EDTA Ethylene diamine tetraacetic acid TCEP Tris(2-carboxyethyl)phosphine hydrochloride DMA Dimethylacetamide DPBS Dulbecco's phosphate buffered saline His / HAc Histidine-acetate buffer HIC Hydrophobic interaction chromatography DETAILED DESCRIPTION OF THE INVENTION

[0089] The following examples are provided to further illustrate the present disclosure but are not intended to limit its scope. Examples

[0090] The structures of the compounds are determined by nuclear magnetic resonance (NMR) or / and mass spectrometry (MS). NMR shifts (5) are given in 10-6 (ppm). NMR measurement is performed using a Bruker AVANCE-400 nuclear magnetic resonance spectrometer, with solvents of deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3) and deuterated methanol (CD3OD), and with an internal standard of tetramethylsilane (TMS).

[0091] MS measurement is conducted using an Agilent 1260 / 1290 Infinity II liquid chromatography system (manufacturer: Agilent; MS model: 1260 / 1290 Infinity II). Shimadzu Prominence UFLC + LCMS -2020 (manufacturer: Shimadzu, MS model: Prominence UFLC + LCMS -2020).

[0092] High performance liquid chromatography (HPLC) analysis is performed using Thermo UltiMate 3000 (manufacturer: Thermo, MS model: UltiMate 3000).

[0093] Chiral HPLC analysis is carried out using a YMC K-PrepLAB100G high-performance liquid chromatograph.

[0094] Preparative high-performance liquid chromatography is performed using Shimadzu LH-40 Liquid Handler, Shimadzu LC-20AP Pump, Shimadzu SPD-20AP UV Detector, and GX-281 Liquid Handler without pump, 4020 Syringe Pump, 333-H3 Pump, 334-H3 Pump, and 1741 UV Detector preparative chromatographs.

[0095] The flash chromatograph used is a Biotage Isolera One flash chromatograph.

[0096] Yantai Xinnuo GF254 silica gel plate is used as the silica gel plate for thin layer chromatography, the specification of the silica gel plate for thin layer chromatography (TLC) is 0.15 mm to 0.2 mm, and the specification for thin layer chromatography for separation and purification of products is 0.4 mm to 0.5 mm.

[0097] Yantai Huanghai silica gel of 200-300 mesh is generally used as the carrier for silica gel column chromatography.

[0098] The average kinase inhibition rate and IC50 value are determined using NovoStar microplate reader (BMG, Germany).

[0099] Known starting materials of the present disclosure may be synthesized using or in accordance with methods known in the art, or may be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, or Chembee Chemicals.

[0100] Unless otherwise specified in the examples, all reactions can be carried out under an argon atmosphere or a nitrogen atmosphere.

[0101] Argon atmosphere or the nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon with a volume of about 1L.

[0102] Hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon with a volume of about 1L.

[0103] The pressurized hydrogenation reaction is performed using a Parr 3916EKX hydrogenator and a Qinglan QL-500 hydrogen generator or an HC2-SS hydrogenator.

[0104] Vacuumization and filling with hydrogen are usually repeatedly operated 3 times prior to the hydrogenation reaction.

[0105] CEM Discover-S 908860 microwave reactor is used for a microwave reaction.

[0106] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0107] Unless otherwise specified in the examples, the reaction temperature is room temperature (20°C to 30°C).

[0108] The reaction progress in the examples is monitored by thin layer chromatography (TLC), the developing agent used in the reaction, the column chromatography eluent system used for purifying the compound and the developing agent system of the thin layer chromatography include: A: dichloromethane / methanol system, and B: n-hexane / ethyl acetate system, the volume ratio of the solvents is adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid can also be added for adjustment. Synthesis of intermediates 1. 2,5-dioxopyrrolidin-1-yl-6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynoate Int-1 Step 1 5-bromo-2-chloro-4-methoxypyrimidine Int-1b

[0109] 5-bromo-2,4-dichloropyrimidine Int-1a (5.0 g, 22 mmol) was dissolved in methanol (60 mL), and a solution of sodium methoxide in methanol (30%, 4.0 g, 22.0 mol) was added to the solution. The reaction mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was concentrated and extracted with ethyl acetate. The organic phase was dried and concentrated to give the compound Int-1b (5.0 g, yield: 100%).

[0110] MS m / z (ESI): 223.1 (M+1)+. Step 2 5-bromo-4-methoxy-2-(methylthio)pyrimidine Int-1c

[0111] To a solution of the compound Int-1b (5.0 g, 22.0 mmol) in N,N-dimethylformamide (25 mL) was added sodium thiomethoxide (1.52 g, 22 mmol), and the reaction mixture was stirred at 40°C for 1 h. After the reaction was completed, the reaction mixture was poured into water and extracted with ethyl acetate (50 mL x 3). The organic phases were combined and dried. The concentrated crude product was purified by silica gel column chromatography with system B to give the compound Int-1c (3.0 g, yield: 60%).

[0112] MS m / z (ESI): 235.0 (M+1)+. Step 3 6-(4-methoxy-2-(methylthio)pyrimidin-5-yl)hex-5-ynoic acid Int-1d

[0113] The compound Int-1c (3.0 g, 12.8 mmol) was dissolved in isopropanol (20 mL), and 5-hexynoic acid (1.43 g, 12.8 mmol), cuprous iodide (243 mg, 1.29 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (300 mg, 0.41 mmol) and sodium carbonate solution (5 M, 8 mL) were added. The reaction mixture was stirred under nitrogen atmosphere at 80°C for 16 h. After the reaction was completed, the reaction mixture was filtered and the filtrate was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, dried and concentrated. The resulting crude product was purified by silica gel column chromatography with system B to give the compound Int-1d (3.0 g, yield: 87%).

[0114] MS m / z (ESI): 267.1 (M+1)+. Step 4 6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynoic acid (Int-1e)

[0115] The compound Int-1d (3.0 g, 11.2 mmol) was dissolved in a mixed solvent of methanol and water (40 mL, V / V=1:1), potassium peroxodisulfate (11.6 g, 33.6 mmol) was added, and the reaction mixture was stirred at room temperature for 3 h. After the reaction was completed, the reaction mixture was poured into water and extracted with ethyl acetate (50 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate to give the crude product. The crude product was purified by silica gel column chromatography with system B to give the compound Int-1e (2.5 g, yield: 75%).

[0116] MS m / z (ESI): 299.1 (M+H)+. Step 5 2,5-dioxopyrrolidin-1-yl-6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynoate Int-1

[0117] To a solution of the compound Int-1e (700 mg, 2.3 mmol) and N-hydroxysuccinimide (396 mg, 3.45 mmol) in tetrahydrofuran (10 mL) was added N,N-diisopropylcarbodiimide (434 mg, 3.45 mmol), and the reaction mixture was stirred at room temperature for 3 h. After the reaction was completed, the reaction mixture was concentrated directly to give the crude product, and the crude product was purified by silica gel column chromatography with system B to give the compound Int-1 (600 mg, yield: 60%).

[0118] MS m / z (ESI): 395.5 (M+1)+. 2. 2,5-dioxopyrrolidin-1-yl-6-(5-cyano-6-(methylsulfonyl)pyridin-3-yl)hex-5-ynoate Int-2 Step 1 5-bromo-2-(methylthio)nicotinonitrile Int-2b

[0119] 5-bromo-2-chloronicotinonitrile Int-2a (2 g, 9.2 mmol) was dissolved in ethylene glycol dimethyl ether (20 mL), and sodium thiomethoxide (640 mg, 9.2 mmol) was added under an ice bath. The mixture was stirred for reaction at room temperature for 4 h. After the reaction was completed, the reaction was quenched with an aqueous ammonium chloride solution, the system was extracted with ethyl acetate three times, and dried over anhydrous sodium sulfate. The organic phases were combined and concentrated. The resulting concentrate was purified by silica gel column chromatography with system B to give the product Int-2b (1.5 g, yield: 71%).

[0120] MS m / z (ESI): 228.9, 230.9 (M+1)+. Step 2 6-(5-cyano-6-(methylthio)pyridin-3-yl)hex-5-ynoic acid Int-2c

[0121] The compound Int-2b (1.5 g, 6.6 mmol) was dissolved in a mixed solvent of tetrahydrofuran (5 mL) and triethylamine (5 mL), and 5-hexynoic acid (1.5 g, 13.2 mmol), bistriphenylphosphine palladium dichloride (913 mg, 1.3 mmol), and cuprous iodide (133 mg, 0.7 mmol) were added thereto. The reaction mixture was allowed to react with stirring at 70°C for 2 h. After the reaction was completed, the reaction mixture was filtered and the mother liquor was directly blended. The resulting crude product was purified by silica gel column chromatography with system B to give the compound Int-2c (0.7 g, yield: 41%).

[0122] MS m / z (ESI): 261.1 (M+1)+. Step 3 6-(5-cyano-6-(methylsulfonyl)pyridin-3-yl)hex-5-ynoic acid Int-2d

[0123] The compound Int-2c (0.7 g, 2.7 mmol) was dissolved in methanol (10 mL) and water (10 mL), and potassium peroxymonosulfate (9.3 g, 27 mmol) was added. The mixture was stirred for reaction at room temperature for two hours. After the reaction was completed, the reaction mixture was filtered, the mother liquor was poured into water, extracted with dichloromethane three times, and dried over anhydrous sodium sulfate. The organic phases were combined and concentrated. The filtrate was collected to give the crude product Int-2d (815 mg), which was directly used in the next reaction step without purification.

[0124] MS m / z (ESI): 293.0 (M+1)+. Step 4 2,5-dioxopyrrolidin-1-yl-6-(5-cyano-6-(methylsulfonyl)pyridin-3-yl)hex-5-ynoate Int-2

[0125] The crude compound Int-2d (815 mg) was dissolved in dichloromethane (10 mL), and N-hydroxysuccinimide (345 mg, 3.0 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (1.04 g, 5.4 mmol) were added and stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was poured into water, extracted with dichloromethane three times, dried over anhydrous sodium sulfate, and the organic phases were combined and concentrated. The resulting concentrate was purified by silica gel column chromatography with system B to give the product Int-2 (400 mg, yield: 37%).

[0126] MS m / z (ESI): 390.0 (M+1)+. 3. 2,5-dioxopyrrolidin-1-yl-6-(4-(methoxymethyl)-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynoate Int-3 Step 1 5-bromo-2-chloro-4-(methoxymethyl)pyrimidine Int-3b

[0127] 5-bromo-2-chloropyrimidine Int-3a (10 g, 0.05 mol), silver nitrate (36 g,0.2 mol), ammonium persulfate (57 g, 0.25 mol) and 2-methoxyacetic acid (5.4 g, 0.06 mol) were dissolved in acetonitrile (300 mL) and water (300 mL) and stirred at 60°C for 2 h. The reaction mixture was poured into water and extracted with ethyl acetate three times, washed once with saturated brine, and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography with system B to give the compound Int-3b (1.2 g, yield: 13%).

[0128] MS m / z (ESI): 236.9 (M+1)+. Step 2 6-(2-chloro-4-(methoxymethyl)pyrimidin-5-yl)hex-5-ynoic acid Int-3c

[0129] The compound Int-3b (1.2 g, 4.5 mmol) was dissolved in tetrahydrofuran (10 mL), and 5-hexynoic acid (0.76 g, 6.8 mmol), cuprous iodide (86 mg, 0.45 mmol), bistriphenylphosphine palladium dichloride (632 mg, 0.9 mmol) and triethylamine (1.4 g, 13.5 mmol) were added. The reaction mixture was stirred under nitrogen atmosphere at 60°C for 3 h. After the reaction was completed, the reaction mixture was filtered, and the filtrate was separated with ethyl acetate and water. The aqueous phase was extracted with ethyl acetate (20 mLx3). The organic phases were combined, dried and concentrated. The resulting crude product was purified by silica gel column chromatography with system A to give the compound Int-3c (600 mg, yield: 50%).

[0130] MS m / z (ESI): 269.1 (M+1)+. Step 3 6-(4-(methoxymethyl)-2-(methylthio)pyrimidin-5-yl)hex-5-ynoic acid Int-3d

[0131] The compound Int-3c (600 mg, 2.2 mmol) was dissolved in dimethyl sulfoxide (6 mL), sodium thiomethoxide (154 mg, 2.2 mmol) and anhydrous magnesium sulfate (528 mg, 4.4 mmol) were added, and the reaction mixture was stirred at 50°C for 1 h. After the reaction was completed, the reaction mixture was poured into water and extracted with ethyl acetate (50 mL x 3). The organic phases were combined, dried, and concentrated to give the crude product Int-3d (600 mg), which was directly used for the next reaction step without purification.

[0132] MS m / z (ESI): 281.1 (M+1)+. Step 4 6-(4-(methoxymethyl)-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynoic acid Int-3e

[0133] The compound Int-3d (600 mg) obtained in the previous step was dissolved in a mixed solvent of acetone and water (20 mL, V / V = 1:1), potassium peroxymonosulfonate (7.6 g, 22 mmol) was added, and the mixture was stirred at room temperature for 1 h. After the reaction was completed, the reaction mixture was poured into water and extracted with ethyl acetate three times, washed once with saturated brine, and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography with system A to give the compound Int-3e (200 mg, yield: 30%).

[0134] MS m / z (ESI): 313.0 (M+1)+. Step 5 2,5-dioxopyrrolidin-1-yl-6-(4-(methoxymethyl)-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynoate Int-3

[0135] The compound Int-3e (200 mg, 0.64 mmol) and N-hydroxysuccinimide (110 mg, 0.96 mmol) were dissolved in dichloromethane (10 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (246 mg, 1.28 mmol) was added, and the reaction was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was concentrated directly to give the crude product, and the crude product was purified by silica gel column chromatography with system B to give the compound Int-3 (100 mg, yield: 38%).

[0136] MS m / z (ESI): 410.1 (M+1)+. Example 1 N-(((S)-1-(((S)-1-([(S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl]amino)-1-oxopropan-2-yl)amino-3-methyl-1-oxobutan-2-yl)-1-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)-3,6,9,12,15,18-hexaoxahenicosan-21-amide (001) Step 1 (9H-fluoren-9-yl)methyl((S)-1-(((S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl(amino)-3-methyl-1-oxobutan-2-yl)carbamate (001b)

[0137] N,N,N',N'-tetramethylchloroformamidinium hexafluorophosphate (333.8 mg, 1.19 mmol) was added to a solution of the compound 001a (400.0 mg, 0.99 mmol), (((9H-fluoren-9- yl)methoxy)carbonyl)-L-valyl-L-alanine (407.0 mg, 0.99 mmol), and N-methylimidazole (170.9 mg, 2.08 mmol) in N,Ndimethylformamide (18.4 mL). The mixture was stirred at 23°C for 3 h. After the reaction was completed, the reaction solution was added dropwise to saturated brine (50 mL), and a yellow solid precipitated out. The system was filtered. The filter cake was dissolved in methanol and dichloromethane, washed twice with water, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the compound 001b (560.0 mg, 71%).

[0138] MS m / z (ESI): 796.2 (M+1). Step 2 (S)-2-amino-N-((S)-1-(((S,9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,2H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)-3-methylbutanamide (001c)

[0139] At 0°C, diethylamine (22.1 mg, 0.30 mmol) was added to a solution of the ompound 001b (80.0 mg, 0.10 mmol) in N,N-dimethylformamide (1 mL). The mixture was stirred at room temperature for 3 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to afford the compound 001c (70.0 mg), which was used in the next step without purification.

[0140] MS m / z (ESI): 574.2 (M+1). Step 3 Tert-butyl-28-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)-23-oxo-4,7,10,13,16,19-hexaoxa-22-azaoctacos-27-ynoate (001e)

[0141] N,N-diisopropylethylamine (98.1 mg, 0.76 mmol) was added to a solution of the compound Int-1 (100.0 mg, 0.25 mmol) and the compound 001d (103.6 mg, 0.25 mmol) in N,N-dimethylformamide (5 mL). The mixture was stirred at room temperature for 2 h. After the reaction was completed, the mixture was added to ethyl acetate (50 mL) and washed twice with water. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford the compound 001e (170.0 mg, 97%).

[0142] MS m / z (ESI): 634.2 (M-56+1). Step 4 28-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)-23-oxo-4,7,10,13,16,19-hexaoxa-22-azaoctacos-27-ynoic acid (001f)

[0143] Trifluoroacetic acid (132.2 mg, 1.16 mmol) was added to a solution of the compound 001e (80.0 mg, 0.12 mmol) in dichloromethane (5 mL) at 0°C, and the mixture was stirred at room temperature for 1 h. After the reaction was completed, the system was concentrated under reduced pressure to afford the compound 001f (80.0 mg), and the product was used in the next step without purification.

[0144] MS m / z (ESI): 634.2 (M+1). Step 5 N-(((S)-1-(((S)-1-([(S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl]amino)-1-oxopropan-2-yl)amino-3-methyl-1-oxobutan-2-yl)-1-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)-3,6,9,12,15,18-hexaoxahenicosan-21-amide (001)

[0145] The compound 001c (57.7 mg, 0.10 mmol) was added to a solution of the compound 001f (70.1 mg, 0.11 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (57.3 mg, 0.15 mmol), and N,N-diisopropylethylamine (39.0 mg, 0.30 mmol) in N,N-dimethylformamide (3 mL). The mixture was stirred at room temperature for 2 h. After completion of the reaction, the mixture was concentrated under reduced pressure. The residue was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector; column: Xtimate C18, 21.2 x 250 mm; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 20%-45%; flow rate: 20 mL / min) to afford the compound 001 (17.3 mg, 14%).

[0146] MS m / z (ESI): 595.5 (M / 2+1).

[0147] 1H NMR (400 MHz, DMSO-d6) 6 9.75 (s, 1H), 8.80 (s, 1H), 8.27 (d, 1H), 7.97 -7.89 (m, 3H), 7.82 (d, 1H), 7.30 (s, 1H), 6.51 (s, 1H), 5.43 (s, 2H), 5.26 (s, 2H), 4.53 (t, 1H), 4.25 (dd, 1H), 4.07 (s, 3H), 3.59 (t, 2H), 3.49 - 3.47 (m, 21H), 3.39 (s, 4H), 3.22 - 3.14 (m, 6H), 2.99 -2.96 (m, 2H), 2.42 - 2.32 (m, 2H), 2.24 (t, 2H), 2.05 - 1.96 (m, 3H), 1.90 - 1.83 (m, 2H), 1.80 -1.75 (m, 2H), 1.39 (d, 3H), 0.90 - 0.84 (m, 9H). Example 2 N-((S)-1-((S)-1-((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-1-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5- ynamido)-3,6,9,12,15,18,21,24-octaoxaheptacosan-27-amide (002) Step 1 Tert-butyl 34-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)-29-oxo- 4,7,10,13,16,19,22,25-octaoxa-28-tetraazanonatriacont-33-ynoate (002b)

[0148] The compound 002a (100 mg, 0.201 mmol) and the compound Int-1 (87 mg, 0.221 mmol) were dissolved in N,N-dimethylformamide (1 mL), and N,N-diisopropylethylamine (78 mg, 0.603 mol) was added dropwise to the reaction system at 0°C. The reaction mixture was heated to 25°C and stirred to react for 2 h. After the reaction was completed, the solvent was removed by rotary evaporation. The residue was dissolved in water (20 mL) and extracted three times with dichloromethane (30 mL). The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and rotary evaporated to dryness to afford the compound 002b (142 mg, yield: 91%), which was used in the next step without purification.

[0149] MS m / z (ESI): 722.3 (M+1-56). Step 2 34-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)-29-oxo-4,7,10,13,16,19,22,25-octaoxa-28-tetraazanonatriacont-33-ynoic acid (002c)

[0150] The compound 002b (60 mg, 0.077 mmol) was dissolved in dichloromethane (1 mL), and then trifluoroacetic acid (0.2 mL, 298 mg, 2.612 mmol) was added at 0 °C. The mixture was then heated to 25°C and stirred for 1 h. After the reaction was completed, the reaction mixture was rotary evaporated to dryness to afford the compound 002c (50 mg), and the product was used in the next step without purification.

[0151] MS m / z (ESI): 722.2 (M+1). Step 3 N-((S)-1-((S)-1-((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-1-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)-3,6,9,12,15,18,21,24-octaoxaheptacosan-27-amide (002)

[0152] The compound 002c (50 mg, 0.069 mmol), HATU (40 mg, 0.104 mmol), and N,N-diisopropylethylamine (27 mg, 0.208 mmol) were dissolved in N,N-dimethylformamide (1 mL). The compound 001c (40 mg, 0.070 mmol) was added at 0°C, and the mixture was stirred to react at 25°C for 2 h. After the reaction was completed, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by preparative high-performance liquid chromatography (Shimadzu LH-40 Liquid Handler, Shimadzu LC-20AP Pump, Shimadzu SPD-20AP UV Detector; column: Xtimate C18, 21.2*250 mm, 5 gm; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 20%-45%; flow rate: 20 mL / min) to afford the compound 002 (22.6 mg, yield: 26%).

[0153] MS m / z (ESI): 639.4 (M / 2+1).

[0154] 1H NMR (400 MHz, DMSO-d6) 5 9.75 (s, 1H), 8.80 (s, 1H), 8.27 (d, 1H), 7.97 - 7.89 (m, 3H), 7.82 (d, 1H), 7.31 (s, 1H), 6.50 (s, 1H), 5.43 (s, 2H), 5.26 (s, 2H), 4.57 - 4.50 (m, 1H), 4.27 - 4.24 (m, 1H), 4.08 (s, 3H), 3.60 (t, 2H), 3.50 - 3.48 (m, 30H), 3.42 - 3.39 (m, 5H), 3.22 - 3.14 (m, 4H), 2.99 - 2.96 (m, 2H), 2.47 - 2.35 (m, 2H), 2.26 - 2.22 (m, 2H), 2.06 - 1.95 (m, 3H), 1.93 - 1.84 (m, 2H), 1.82 - 1.74 (m, 2H), 1.40 (d, 3H), 0.90 - 0.84 (m, 9H). Example 3 N-(((S)-1-(((S)-1-([(S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl]amino)-1-oxopropan-2-yl)amino-3-methyl-1-oxobutan-2-yl)-1-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-amide (003) Step 1 Tert-butyl 40-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)-35-oxo-4,7,10,13,16,19,22,25,28,31-decaoxa-34-tetraazatritetracont-39-ynoate (003b)

[0155] The compound 003a (100 mg, 0.17 mmol) and the compound Int-1 (67.5 mg, 0.17 mmol) were dissolved in N,N-dimethylformamide (5 mL), N,N-diisopropylethylamine (66.1 mg, 0.51 mmol) was added, and the mixture was stirred at room temperature for 2 h. After the reaction was completed, water and ethyl acetate were added for extraction. The organic phase was washed twice with saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give the crude product, which was purified by silica gel column chromatography with system A to afford the compound 003b (110 mg, 74.4%).

[0156] MS m / z (ESI): 886.4 (M+1). Step 2 40-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)-35-oxo-4,7,10,13,16,19,22,25,28,31-decaoxa-34-tetraazatritetracont-39-ynoic acid (003c)

[0157] The compound 003b (100 mg, 0.17 mmol) was dissolved in dichloromethane (2 mL). The solution was cooled to 0°C, trifluoroacetic acid (0.4 mL) was added slowly, and the mixture was stirred at room temperature for 1 h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to give the crude compound 003c (90 mg), which was directly used in the next reaction step without purification.

[0158] MS m / z (ESI): 810.4 (M+1). Step 3 N-(((S)-1-(((S)-1-([(S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl]amino)-1-oxopropan-2-yl)amino-3-methyl-1-oxobutan-2-yl)-1-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-amide 003)

[0159] The compound 003c (80 mg, 0.10 mmol) and the compound 001c (56.7 mg, 0.10 mmol) were dissolved in N,N-dimethylformamide (2 mL), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (56.3 mg, 0.15 mmol) and N,N-diisopropylethylamine (38.2 mg, 0.30 mmol) were added, and the mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was filtered. The crude product was purified by preparative high-performance liquid chromatography (Shimadzu LH-40 Liquid Handler, Shimadzu LC-20AP Pump, Shimadzu SPD-20AP UV Detector; column: Xtimate 5 pm C18 250 x 21.2 mm; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 21-min gradient, gradient ratio: acetonitrile phase 20%-48%; flow rate: 20 mL / min) to afford the compound 003 (24.1 mg, 17.9%).

[0160] MS m / z (ESI): 683.4 (M / 2+1).

[0161] 1H NMR (400 MHz, DMSO-d6) 5 9.78 (s, 1H), 8.82 (s, 1H), 8.29 (d, 1H), 8.02 - 7.81 (m, 4H), 7.33 (s, 1H), 6.53 (s, 1H), 5.45 (s, 2H), 5.28 (s, 2H), 4.55 (t, 1H), 4.31 - 4.22 (m, 1H), 4.09 (s, 3H), 3.61 (t, 2H), 3.51 - 3.50 (m, 34H), 3.43 - 3.41 (m, 5H), 3.24 - 3.18 (m, 5H), 3.01 - 2.98 (m, 2H), 2.70 - 2.68 (m, 1H), 2.28 - 2.24 (m, 2H), 2.09 - 1.96 (m, 4H), 1.93 - 1.76 (m, 5H), 1.41 (d, 3H), 1.25 (s, 2H), 0.92 - 0.86 (m, 9H). Example 4 1-(6-(5-cyano-6-(methylsulfonyl)pyridin-3-yl)hex-5-ynamido)-N-((S)-1-(((S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H- benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1 -oxobutan-2-yl)-3,6,9,12,15,18-hexaoxahenicosan-21 -amide (004)

[0162] Following the synthetic route of Example 1, the starting material in step 1 was replaced with Int-2. The crude product was purified by preparative high-performance liquid chromatography (Shimadzu LH-40 Liquid Handler, Shimadzu LC-20AP Pump, Shimadzu SPD-20AP UV Detector; column: Xtimate C18, 21.2*250 mm, 5 pm; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 20%-45%; flow rate: 20 mL / min), and freeze-dried to afford the compound 004 (10.7 mg, 17%).

[0163] MS m / z (ESI): 592.4 (M+2 / 2).

[0164] 1H NMR (400 MHz, DMSO-d6) 8 9.82 (s, 1H), 9.04 (d, 1H), 8.78 (d, 1H), 8.33 (d, 1H), 8.04 - 7.94 (m, 3H), 7.88 (d, 1H), 7.37 (s, 1H), 6.56 (s, 1H), 5.49 (s, 2H), 5.32 (s, 2H), 4.62 -4.56 (m, 1H), 4.33 - 4.29 (m, 1H), 3.67 - 3.64 (m, 2H), 3.57 - 3.53 (m, 20H), 3.51 (s, 3H), 3.49 - 3.45 (m, 4H), 3.28 - 3.26 (m, 2H), 3.24 - 3.21 (m, 2H), 3.05 - 3.02 (m, 2H), 2.48 - 2.39 (m, 2H), 2.34 - 2.31 (m, 2H), 2.08-2.03 (m, 2H) 1.99-1.91 (m, 2H)1.86 - 1.82 (m, 2H), 1.46 (d, 3H), 0.97 - 0.89 (m, 9H). Example 5 1-(6-(5-cyano-6-(methylsulfonyl)pyridin-3-yl)hex-5-ynamido)-N-((S)-1-(((S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-3,6,9,12,15,18,21,24-octaoxaheptacosan-27-amide (005)

[0165] Following the synthetic route of Example 2, the starting material in step 1 was replaced with Int-2. The crude product was purified by preparative high-performance liquid chromatography (Shimadzu LH-40 Liquid Handler, Shimadzu LC-20AP Pump, Shimadzu SPD-20AP UV Detector; column: Xtimate C18, 21.2*250 mm, 5 pm; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 20%-46%; flow rate: 20 mL / min), and freeze-dried to afford the compound 005 (10.7 mg, 13.7%).

[0166] MS m / z (ESI): 1271.4 (M+1).

[0167] 1H NMR (400 MHz, DMSO-d6) 8 9.76 (s, 1H), 8.98 (d, 1H), 8.72 (d, 1H), 8.27 (d, 1H), 7.99 - 7.90 (m, 3H), 7.83 (d, 1H), 7.31 (s, 1H), 6.51 (s, 1H), 5.44 (s, 2H), 5.27 (s, 2H), 4.55 -4.52 (m, 1H), 4.27 - 4.24 (m, 1H), 3.61 - 3.58 (m, 2H), 3.50 (s, 28H), 3.45 (s, 3H), 3.42 - 3.39 (m, 4H), 3.23 - 3.21 (m, 2H),3.18-3.16 (m, 2H) 2.99 - 2.97 (m, 2H), 2.42-2.33 (m, 2H), 2.29 -2.25 (m, 2H), 2.02 - 1.97 (m, 3H), 1.91 - 1.84 (m, 2H), 1.82-1.76 (m, 2H)1.40 (d, 3H), 0.91 -0.83 (m, 9H). Example 6 1-(6-(5-cyano-6-(methylsulfonyl)pyridin-3-yl)hex-5-ynamido)-N-((S)-1-(((S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H- benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-amide (006)

[0168] Following the synthetic route of Example 3, the starting material in step 1 was replaced with Int-2. After the reaction was completed, the reaction mixture was directly purified by preparative high-performance liquid chromatography (Shimadzu LH-40 Liquid Handler, Shimadzu LC-20AP Pump, Shimadzu SPD-20AP UV Detector; column: Xtimate C18, 21.2*250 mm, 5 gm; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 21%-46%; flow rate: 20 mL / min) to afford the compound 006 (11.2 mg, yield: 18%).

[0169] MS m / z (ESI): 1360.5 (M+1).

[0170] 1H NMR (400 MHz, CD3OD) 5 8.85 (d, 1H), 8.47 (d, 1H), 7.97 (d, 1H), 7.82 (d, J = 9.2 Hz, 1H), 7.66 (s, 1H), 5.62 (d, 1H), 5.42(d, 1H), 5.26 (s, 2H), 4.72 - 4.58 (m, 2H), 4.27 (d, 1H), 3.77 -3.74 (m, 2H),3.63 -3.61 (m, 38H) 3.56-3.53 (m, 2H), 3.39-3.33 (m, 4H), 3.23-3.20 (m, 2H), 3.08-3.04 (m, 2H), 2.62 - 2.55 (m, 4H), 2.43-2.39 (m, 2H), 2.21 - 2.12 (m, 3H), 2.03 - 1.92 (m, 4H), 1.58-1.56 (m, 3H), 1.05-1.00 (m, 9H). Example 7 N-(((S)-1-(((S)-1-([(S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-1-(6-(4-(methoxymethyl)-2-(methylsulfonyl)pyrimidin-5- yl)hex-5-ynamido)-3,6,9,12,15,18-hexaoxahenicosan-amide (007)

[0171] Following the synthetic route of Example 1, the starting material in step 1 was replaced with Int-3. The crude product was concentrated under reduced pressure. The residue was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector; column: Xtimate C18, 21.2 x 250 mm; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 20%-45%; flow rate: 20 mL / min) to afford the compound 007 (32.0 mg, 26%).

[0172] MS m / z (ESI): 602.5 (M / 2+1).

[0173] 1H NMR (400 MHz, DMSO-d6) 5 9.75 (s, 1H), 9.04 (s, 1H), 8.27 (d, 1H), 7.97 - 7.90 (m, 3H), 7.82 (d, 1H), 7.30 (s, 1H), 6.50 (s, 1H), 5.43 (s, 2H), 5.26 (s, 2H), 4.70 (s, 2H), 4.53 (t, 1H), 4.25 (dd, 1H), 3.59 (t, 2H), 3.49 - 3.47 (m, 20H), 3.41 - 3.38 (m, 8H), 3.22 - 3.14 (m, 4H), 2.97 (t, 2H), 2.57 (t, 2H), 2.46 - 2.35 (m, 2H), 2.27 (t, 2H), 2.06 - 1.96 (m, 3H), 1.91 - 1.78 (m, 4H), 1.39 (d, 3H), 0.90 - 0.84 (m, 9H). Example 8 N-((S)-1-((S)-1-((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-1-(6-(4-(methoxymethyl)-2-(methylsulfonyl)pyrimidin-5-yl)hexyl)-3,6,9,12,15,18,21,24-octaoxaheptacosan-27-amide (008)

[0174] Following the synthetic route of Example 2, the starting material in step 1 was replaced with Int-3. After the reaction was completed, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by preparative high-performance liquid chromatography (Shimadzu LH-40 Liquid Handler, Shimadzu LC-20AP Pump, Shimadzu SPD-20AP UV Detector; column: Xtimate C18, 21.2*250 mm, 5 gm; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 20%-45%; flow rate: 20 mL / min) to afford the compound 008 (18.1 mg, yield: 21%).

[0175] MS m / z (ESI): 646.4 (M / 2+1).

[0176] 1H NMR (400 MHz, DMSO) 8 9.76 (s, 1H), 9.05 (s, 1H), 8.28 (d, 1H), 7.97 - 7.91 (m, 3H), 7.83 (d, 1H), 7.31 (s, 1H), 6.51 (s, 1H), 5.43 (s, 2H), 5.26 (s, 2H), 4.71 (s, 2H), 4.57 - 4.50 (m, 1H), 4.28 - 4.24 (m, 1H), 3.60 (t, 2H), 3.50 - 3.48 (m, 28H), 3.42 - 3.39 (m, 8H), 3.23 - 3.14 (m, 4H), 2.98 (t, 2H), 2.58 (t, 2H), 2.49 - 2.36 (m, 2H), 2.28 (t, 2H), 2.08 - 1.96 (m, 3H), 1.93 - 1.79 (m, 4H), 1.40 (d, 3H), 0.91 - 0.85 (m, 9H). Example 9 N-(((S)-1-(((S)-1-((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1, 2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-1-(6-(4-(methoxymethyl)-2-(methylsulfonyl)pyrimidin-5- yl)hex-5-ynamido)-3,6,9,12,15,18,21,24,27,30-decaoxatritriacontan-33-amide (009)

[0177] Following the synthetic route of Example 3, the starting material in step 1 was replaced with Int-3. The crude product was purified by preparative high-performance liquid chromatography (Shimadzu LH-40 Liquid Handler, Shimadzu LC-20AP Pump, Shimadzu SPD-20AP UV Detector; column: Xtimate 5 ^m C18 250 x 21.2 mm; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 20%-48%; flow rate: 20 mL / min) to afford the compound 009 (23.2 mg, 19.8%).

[0178] MS m / z (ESI): 690.4 (M / 2+1).

[0179] 1H NMR (400 MHz, DMSO-d6) 5 9.78 (s, 1H), 9.07 (s, 1H), 8.29 (d, 1H), 7.99 - 7.83 (m, 4H), 7.33 (s, 1H), 6.53 (s, 1H), 5.45 (s, 2H), 5.29 (s, 2H), 4.72 (s, 2H), 4.57 - 4.53 (m, 1H), 4.29 - 4.25 (m, 1H), 3.61 (t, 2H), 3.52 - 3.50 (m, 34H), 3.43 - 3.42 (m, 7H), 3.21 - 3.18 (m, 4H), 3.01 - 2.98 (m, 2H), 2.70 - 2.68 (s, 1H), 2.62 - 2.56 (m, 3H), 2.36 - 2.27 (m, 3H), 2.08 - 1.98 (m, 3H), 1.91 - 1.82 (m, 4H), 1.41 (d, 3H), 1.27 - 1.24 (m, 2H), 0.92 - 0.86 (m, 9H). Example 10 N-(2-(3-(((S)-1-(((S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethyl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)-N-(2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-28-yl)hex-5-ynamide Step 1 2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-28-al 011b

[0180] 011a (500.0 mg, 1.17 mmol) and Dess-Martin periodinane (742.3 mg, 1.75 mmol) were dissolved in dichloromethane (10 mL) and stirred at room temperature for 18 h. After the reaction was completed, the mixture was washed with saturated sodium bicarbonate solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to afford 011b as a colorless oil (400.0 mg, 80%).

[0181] MS m / z (ESI): 427.4 (M+1). Step 2 Tert-butyl 2,5,8,11,14,17,20,23,26,32-decaoxa-29-aza-32-oxapentatriacontan-35-oate 011c

[0182] 011b (100.0 mg, 0.23 mmol), tert-butyl 3-(2-aminoethoxy)propanoate (39.9 mg, 0.21 mmol), and acetic acid (6.9 mg, 0.12 mmol) were dissolved in methanol (5 mL) and stirred at room temperature for 30 min. Sodium cyanoborohydride (29.5 mg, 0.47 mmol) was added, the system was purged with nitrogen three times, and the mixture was stirred at room temperature for 18 h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with system A to afford 011c as a colorless oil (35.0 mg, 25%).

[0183] MS m / z (ESI): 600.4 (M+1). Step 3 Tert-butyl 29-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynyl)-2,5,8,11,14,17,20,23,26,32-decaoxa-29-aza-32-oxapentatriacontan-35-oate 011d

[0184] 011c (35.0 mg, 0.06 mmol), 2,5-dioxopyrrolidin-1-yl 6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynoate Int-1 (23.1 mg, 0.06 mmol), and N,N-diisopropylethylamine (22.6 mg, 0.18 mmol) were dissolved in N,N-dimethylformamide (2 mL), heated to 50°C, and stirred for 18 h. After the reaction was complete, ethyl acetate and water were added for extraction. The organic phase was concentrated under reduced pressure, and the residue was purified by column chromatography with system A to afford 011d as a colorless oil (17.0 mg, 33%).

[0185] MS m / z (ESI): 880.5 (M+1). Step 4 29-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynyl)-2,5,8,11,14,17,20,23,26,32-decaoxa-29-aza-32-oxapentatriacontan-35-oic acid 011e

[0186] 011d (17.0 mg, 0.02 mmol) was dissolved in dichloromethane (2 mL). Trifluoroacetic acid (44.1 mg, 0.39 mmol) was added at 0°C, and the mixture was stirred at room temperature for 2 h. After the reaction was completed, the mixture was concentrated under reduced pressure to afford crude 011e (20.0 mg). The product was used directly in the next step without purification.

[0187] MS m / z (ESI): 824.3 (M+1). Step 5 N-(2-(3-(((S)-1-(((S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethyl)-6-(4-methoxy-2- (methylsulfonyl)pyrimidin-5-yl)-N-(2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-28-yl)hex-5-ynamide 011

[0188] The compound 011e (20.0 mg, 0.020 mmol), the compound 001c (11.5 mg, 0.02 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11.4 mg, 0.03 mmol) were dissolved in N,N-dimethylformamide (2 mL). N,N-diisopropylethylamine (7.8 mg, 0.06 mmol) was added, and the mixture was stirred at room temperature for 18 h. After the reaction was completed, the reaction mixture was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector; column: Xtimate C18, 21.2 x 250 mm; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 20%-45%; flow rate: 20 mL / min) to afford the compound 011 (3.4 mg, 12%).

[0189] MS m / z (ESI): 690.6 (M / 2+1).

[0190] 1H NMR (400 MHz, DMSO-d6) 5 9.75 (s, 1H), 8.81 - 8.79 (m, 1H), 8.27 (d, 1H), 7.95 (d, 1H), 7.94 - 7.90 (m, 1H), 7.82 (d, 1H), 7.30 (s, 1H), 6.50 (s, 1H), 5.43 (s, 2H), 5.26 (s, 2H), 4.57 - 4.50 (m, 1H), 4.28 - 4.23 (m, 1H), 4.07 (d, 3H), 3.61 - 3.54 (m, 2H), 3.54 - 3.46 (m, 29H), 3.46 - 3.39 (m, 6H), 3.38 (s, 3H), 3.35 - 3.34 (m, 1H), 3.30 - 3.26 (m, 2H), 3.23 (s, 3H), 3.18 - 3.12 (m, 2H), 3.00 - 2.94 (s, 2H), 2.56 - 2.52 (m, 3H), 2.48 - 2.43 (m, 2H), 2.40 - 2.32 (m, 2H), 2.06 - 1.94 (m, 4H), 1.94 - 1.83 (m, 2H), 1.81 - 1.76 (m, 2H), 1.39 (d, 3H), 0.90 - 0.84 (m, 9H). Example 11 N-(4-(((S)-1-(((S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-4-oxobutyl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)-N-(2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-28-yl)hex-5-ynamide 012 Step 1 Tert-butyl 2,5,8,11,14,17,20,23,26-nonaoxa-29-azatritriacontan-33-oate 012a

[0191] 011a (100.0 mg, 0.23 mmol), tert-butyl 4-aminobutanoate hydrochloride (41.3 mg, 0.21 mmol), and acetic acid (6.9 mg, 0.12 mmol) were dissolved in methanol (5 mL) and stirred at room temperature for 30 min. Sodium cyanoborohydride (29.5 mg, 0.47 mmol) was added, the system was purged with nitrogen three times, and the mixture was stirred at room temperature for 18 h. After the reaction was completed, the reaction mixture was purified by column chromatography with system A to afford the compound 012a (77.0 mg, 58%).

[0192] MS m / z (ESI): 570.4 (M+1). Step 2 Tert-butyl 29-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynyl)-2,5,8,11,14,17,20,23,26-nonaoxa-29-azatritriacontan-33-oate 012b

[0193] The compound 012a (77.0 mg, 0.14 mmol), 2,5-dioxopyrrolidin-1-yl-6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynoate Int-1 (53.4 mg, 0.14 mmol), and N,N-diisopropylethylamine (52.4 mg, 0.41 mmol) were dissolved in N,N-dimethylformamide (5 mL), heated to 50°C, and stirred for 18 h. After the reaction was completed, ethyl acetate and water were added for extraction. The organic phase was concentrated under reduced pressure, and the residue was purified by column chromatography with system A to afford the compound 012b (60.0 mg, 52%).

[0194] MS m / z (ESI): 850.4 (M+1). Step 3 29-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynyl)-2,5,8,11,14,17,20,23,26-nonaoxa-29-azatritriacontan-33-oic acid 012c

[0195] 012b (60.0 mg, 0.07 mmol) was dissolved in dichloromethane (2 mL). Trifluoroacetic acid (161.0 mg, 1.41 mmol) was added at 0°C, and the mixture was stirred at room temperature for 2 h. After the reaction was completed, the mixture was concentrated under reduced pressure to afford crude 012c (65 mg). The product was used directly in the next step without purification.

[0196] MS m / z (ESI): 794.2 (M+1). Step 4 N-(4-(((S)-1-(((S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-4-oxobutyl)-6-(4-methoxy-2- (methylsulfonyl)pyrimidin-5-yl)-N-(2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-28-yl)hex-5-ynamide 012

[0197] The compound 012c (60.0 mg, 0.07 mmol), the compound 001c (40.5 mg, 0.07 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (40.3 mg, 0.11 mmol) were dissolved in N,N-dimethylformamide (2 mL). N,Ndiisopropylethylamine (27.4 mg, 0.21 mmol) was added, and the mixture was stirred at room temperature for 18 h. After the reaction was completed, the reaction mixture was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector; column: Xtimate C18, 21.2 x 250 mm; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 20%-45%; flow rate: 20 mL / min) to afford the compound 012 (29.9 mg, 31%).

[0198] MS m / z (ESI): 675.6 (M / 2+1)o

[0199] 1H NMR (400 MHz, DMSO-d6) 6 9.73 (d, 1H), 8.79 (s, 1H), 8.27 (d, 1H), 7.95 (d, 1H), 7.92 - 7.89 (m, 1H), 7.81 (d, 1H), 7.30 (s, 1H), 6.50 (s, 1H), 5.43 (s, 2H), 5.26 (s, 2H), 4.54 -4.51 (m, 1H), 4.26 - 4.18 (m, 1H), 4.07 (d, 3H), 3.51 - 3.47 (m, 32H), 3.43 -3.40 (m, 4H), 3.38 (s, 3H), 3.23 (s, 3H), 3.18 -3.18 (m, 2H), 3.00 - 2.93 (m, 2H), 2.56 -2.54 (m, 3H), 2.46 -2.43 (m, 2H), 2.23 - 2.12 (m, 2H), 2.07 - 2.12 (m, 4H), 1.92 - 1.83 (m, 2H), 1.82 - 1.73 (m, 3H), 1.68 - 1.64 (m, 1H), 1.39 (d, 3H), 0.90 - 0.84 (m, 9H). Example 12 N-((5S,14S)-5-benzyl-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-14-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)-1,4,7,10,13-pentaoxo-3,6,9,12-tetraazaoctadecan-18-yl)-2,5,8,11,14,17,20,23-octaoxahexacosanamide 018

[0200] The synthetic route of Example 13 was followed. The residue was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with QDA detector; column: Xtimate C18, 21.2 x 250 mm; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 20%-45%; flow rate: 20 mL / min) to afford the compound 018 (5.8 mg, 15.8%).

[0201] MS m / z (ESI): 762.5 (M / 2+1)+.

[0202] 1H NMR (400 MHz, DMSO-d6) 8 9.64 (s, 1H), 8.79 (s, 1H), 8.52 - 8.46 (m, 1H), 8.24 -8.16 (m, 2H), 8.04 - 8.02 (m, 2H), 7.99 - 7.96 (m, 1H), 7.90 - 7.86 (m, 1H), 7.79 - 7.77 (m, 1H), 7.32 (s, 1H), 7.27 - 7.25 (m, 3H), 7.23 - 7.15 (m, 2H), 6.51 (s, 1H), 5.44 (s, 2H), 5.27 (s, 2H), 4.60 - 4.54 (m, 1H), 4.24 - 4.17 (m, 1H), 4.07 (s, 3H), 4.03 - 3.99 (m, 2H), 3.78 - 3.66 (m, 4H), 3.59 - 3.55 (m, 2H), 3.50 - 3.47 (m, 26H), 3.25 - 3.21 (m, 5H), 3.19 - 3.14 (m, 4H), 3.01 - 2.96 (m, 5H), 2.88 - 2.82 (m, 2H), 2.31 - 2.27 (m, 4H), 2.07 - 2.01 (m, 2H), 1.90 - 1.86 (m, 2H), 1.80 - 1.76 (m, 2H), 1.72 - 1.57 (m, 2H), 1.54 - 1.41 (m, 2H), 1.38 - 1.32 (m, 2H), 1.27 - 1.23 (m, 2H), 0.89 (t, 3H). Example 13 N-((5S,14S)-5-benzyl-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-14-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)-1,4,7,10,13-pentaoxo-3,6,9,12- tetraazaoctadecan-18-yl)-2,5,8,11,14,17,20,23,26,29,35-dodecaoxatetratriacontanamide 019 Step 1 (9H-fluoren-9-yl)methyl (2-((((S)-1-((2-([(((R)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-2-oxoethyl)amino)-1-oxo-3-phenylpropan-2-yl)amino (019a)

[0203] 001 (100 mg, 0.248 mmol), (((9H-fluoren-9-yl)methoxy)carbonyl)glycylglycyl-L-phenylalanylglycine (138.46 mg, 0.248 mmol), and 1-methylimidazole (42.73 mg, 0.521 mmol) were added to DMF (5 mL). N,N,N',N'-tetramethylchloroformamidinium hexafluorophosphate (83.46 mg, 0.297 mmol) was then added. The reaction mixture was stirred at room temperature under nitrogen protection for 18 h. The mixture was poured into saturated aqueous sodium chloride solution (25 mL) and filtered. The yellow solid was dissolved in DCM / CH3OH. The organic layer was washed twice with H2O and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure to give the crude product 019a (132 mg, 56.4%).

[0204] MS m / z (ESI): 943.6 (M+1)+. Step 2 (S)-2-(2-(2-aminoacetamido)acetamido)-N-(2-(((R)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-2-oxoethyl)-3-phenylpropanamide (019b)

[0205] At 20°C, diethylamine (0.087 mL, 0.839 mmol) was added to a solution of 019a (132 mg, 0.140 mmol) in DMF (2 mL). The mixture was then stirred at 20°C for 1 h. The residue was purified by HPLC (TFA, H2O:MeCN = 57%) to give the product 019b (76 mg, 75.31%).

[0206] MS m / z (ESI): 721.6 (M+1)+. Step 3 Tert-butyl (S)-44-(((9H-fluoren-9-yl)methoxy)carbonylamino)-38-oxo-2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxa-39-tetraazaoctatetracontan-45-oate (019d)

[0207] At 25°C, to a solution of 019c (250 mg, 0.589 mmol) in DMF (4ml) were added N,N-diisopropylethylamine (0.195 mL, 1.178 mmol), HATU (291.09 mg, 0.766 mmol), and undecaethylene glycol monomethyl ether propanoic acid (346.66 mg, 0.589 mmol). The mixture was stirred at 25°C under nitrogen protection for 18 h. After the reaction was completed, the mixture was partitioned between ethyl acetate (20 mL) and water (20 mL). The aqueous layer was extracted with ethyl acetate (20 mL * 2), the combined organic layer was dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography with system A to give the product 019d (467 mg, 79.69%).

[0208] MS m / z (ESI): 995.4 (M+1)+. Step 4 Tert-butyl (S)-44-amino-38-oxo-2,5,8,11,14,17,20,23,26,29,35-dodecaoxa-39-tetraazacyclopentatetracontanoate 019e

[0209] To a solution of 019d (417 mg, 0.419 mmol) in DCM (2 mL) was added diethylamine (0.087 mL, 0.838 mmol) at 25°C. The reaction mixture was then stirred at 25°C for 1 h. The solvent was removed under reduced pressure to afford 019e (310 mg, 95.7%). The product was used directly in the next step without purification.

[0210] MS m / z (ESI): 773.6 (M+1)+. Step 5 Tert-butyl (S)-44-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)-38-oxo-2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxa-39-tetraazaoctatetracontan-45-oate 019f

[0211] To a solution of 019e (310.0 mg, 0.401 mmol) in DMF (5 mL) were added N,N-diisopropylethylamine (0.133 mL, 0.802 mmol) and Int 1 (158.57 mg, 0.40 mmol) at 25°C. The reaction mixture was then stirred at 25°C for 2 h. The residue was purified by reversed-phase column chromatography to give the title product 019f (267 mg, 63.2%) as a colorless oil.

[0212] MS m / z (ESI): 1052.6 (M+1)+. Step 6 (S)-44-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)-38-oxo-2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxa-39-tetraazaoctatetracontenoic acid (019g)

[0213] 019f (110 mg, 0.104 mmol) was added to a mixed solution of DCM (1 mL) and TFA (0.2 mL) at 20°C. The reaction mixture was then stirred at 20°C for 2 h. The reaction solution was concentrated under vacuum to give the crude product 019g (104 mg, 99.8%). The crude product was directly used in the next reaction step without purification.

[0214] MS m / z (ESI): 996.6 (M+1). Step 7 N-((5S,14S)-5-benzyl-1-(((R)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-14-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)-1,4,7,10,13-pentaoxo-3,6,9,12-tetraazaoctadecan-18-yl)-2,5,8,11,14,17,20,23,26,29,35-dodecaoxaoctatriacontan-38-amide (019)

[0215] To a solution of 019b (70 mg, 0.097 mmol) in DMF (1 mL) were added N,N-diisopropylethylamine (0.048 mL, 0.291 mmol), 019g (106.38 mg, 0.107 mmol), and HATU (55.32 mg, 0.145 mmol) at 0°C. The mixture was then stirred at 20°C for 1 h. After the reaction was completed, the reaction mixture was directly purified by preparative high-performance liquid chromatography (GX-281 Liquid Handler without pump, 4020 Syringe pump, 333-H3 Pump, 334-H3 Pump, 1741 U; column: Bonnasil-BS C18,20*250 mm,8um; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 20%-50%; flow rate: 20 mL / min) to afford the title compound 019 (16 mg, 0.009 mmol, 9.70%).

[0216] MS m / z (ESI): 1700.7 (M+1)+.

[0217] 1H NMR (400 MHz, DMSO-d6) 5 9.64 (s, 1H), 8.79 (s, 1H), 8.49 (d, 1H), 8.22 - 8.15 (m, 2H), 8.04 (d, 2H), 7.97 (d, 1H), 7.87 (d, 1H), 7.80 - 7.76 (m, 1H), 7.31 (s, 1H), 7.29 - 7.24 (m, 4H), 7.19 (dd, 1H), 6.52 (s, 1H), 5.44 (s, 2H), 5.27 (s, 2H), 4.57 (dd, 1H), 4.26 - 4.16 (m, 1H), 4.07 (s, 3H), 4.04 - 3.88 (m, 3H), 3.83 - 3.60 (m, 5H), 3.60 - 3.54 (m, 3H), 3.50 - 3.38 (m, 26H), 3.33 (s, 7H), 3.26 - 3.06 (m, 9H), 2.99 (d, 5H), 2.85 (dd, 2H), 2.54 (d, 2H), 2.32 - 2.24 (m, 5H), 2.04 (s, 2H), 1.95 - 1.72 (m, 5H), 1.71 - 1.44 (m, 3H), 1.43 - 1.03 (m, 6H), 0.92 - 0.84 (m, 3H). Example 14 6-(4-((2,5,8,11,14-pentaoxahexadecan-16-yl)oxy)-2-(methylsulfonyl)pyrimidin-5-yl)-N-((2S,5S)-1-((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-5-isopropyl-2-methyl-1,4,7- trioxo-10,13-dioxa-3,6-diazapentadecan-15-yl)hex-5-ynamide 028 Step 1: Methyl 6-(2,4-dichloropyrimidin-5-yl)hexanoate (028b)

[0218] The compound 028a (6000 mg, 21.669 mmol), methyl 5-hexynoate (4100 mg, 32.504 mmol), triethylamine (6.0 mL, 43.339 mmol), cuprous iodide (825 mg, 4.334 mmol), and bistriphenylphosphine palladium dichloride (1686 mg, 2.167 mmol) were dissolved in THF (25 mL) and stirred to react at 70°C for 2 h. After the reaction was completed, H2O (30 mL) was added. The mixture was extracted with EA (50mL x3). The organic phase was washed with brine, dried over anhydrous Na2SO4, concentrated under reduced pressure, and purified by column chromatography (PE:EA=10 / 1) to afford the compound 028b (3.6 g, yield: 61%).

[0219] MS m / z (ESI): 272.6 (M+1). Step 2 Methyl 6-(4-((2,5,8,11,14-pentaoxahexadecan-16-yl)oxy)-2-chloropyrimidin-5-yl)hexanoate (028c)

[0220] The compound 028b (1100 mg, 4.028 mmol), 2,5,8,11,14-pentaoxahexadecan-16-ol (1016 mg, 4.028 mmol), and potassium carbonate (1113 mg, 8.056 mmol) were dissolved in DMF (6 mL) and stirred at 70°C under nitrogen protection for 18 h. The reaction mixture was purified by column chromatography with system A to afford the compound 028c (600 mg, yield: 30%).

[0221] MS m / z (ESI): 489.0 (M+1). Step 3 Methyl 6-(4-((2,5,8,11,14-pentaoxahexadecan-16-yl)oxy)-2-(methylthio)pyrimidin-5-yl)hexanoate (028d)

[0222] The compound 028c (400 mg, 0.818 mmol) and sodium thiomethoxide (114 mg, 1.636 mmol) were dissolved in DMF (4 mL) and stirred at 25°C under nitrogen for 2 h. The reaction mixture was purified by column chromatography with system A to afford the compound 028d (200 mg, yield: 49%).

[0223] MS m / z (ESI): 501.1 (M+1). Step 4 6-(4-((2,5,8,11,14-Pentaoxahexadecan-16-yl)oxy)-2-(methylthio)pyrimidin-5-yl)hexanoic acid (028e)

[0224] The compound 028d (300 mg, 0.599 mmol) and LiOH (50 mg, 1.198 mmol) were dissolved in THF (3 mL) / H2O (1 mL) and stirred at 25°C under nitrogen for 2 h. The reaction mixture was purified by column chromatography with system A to afford the compound 028e (270 mg, yield: 93%).

[0225] MS m / z (ESI): 508.7 (M+23)+. Step 5 6-(4-((2,5,8,11,14-pentaoxahexadecan-16-yl)oxy)-2-(methylsulfonyl)pyrimidin-5-yl)hexanoic acid (028f)

[0226] The compound 028e (250 mg, 0.514 mmol) was dissolved in THF (5 mL). Metachloroperoxybenzoic acid (354 mg, 2.055 mmol) was slowly added at 0°C, and the reaction mixture was stirred at 0°C for 3 h. The reaction mixture was purified by column chromatography with system A to afford the compound 028f (245 mg, yield: 92%).

[0227] MS m / z (ESI): 519.2 (M+1)+. Step 6 Tert-butyl 3-(2-(2-(6-(4-((2,5,8,11,14-pentaoxahexadecan-16-yl)oxy)-2-(methylsulfonyl)pyrimidin-5-yl)hexadecyl)ethoxy)propanoate (028g)

[0228] The compound 028f (50 mg, 0.096 mmol), HATU (55 mg, 0.145 mmol), N,N-diisopropylethylamine (37 mg, 0.289 mmol) were dissolved in N,N-dimethylformamide (1 mL), tert-butyl 3-(2-(2-aminoethoxy)ethoxy)propanoate (25 mg, 0.106 mmol) was added at 0°C, and the reaction mixture was stirred at 25°C for 18 h. After the reaction was completed, most of the solvent was removed by sprin-drying. Water (10 mL) was added, and the mixture was extracted with dichloromethane (20 mL) three times. The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and rotary evaporated to dryness to afford the crude compound 028g (70 mg). The product was used directly in the next step without purification.

[0229] MS m / z (ESI): 734.3 (M+1)+. Step 7 3-(2-(2-(6-(4-((2,5,8,11,14-pentaoxahexadecan-16-yl)oxy)-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)ethoxy)propanoic acid (028h)

[0230] The compound 028g (70 mg, 0.095 mmol) was dissolved in dichloromethane (1 mL). Trifluoroacetic acid (0.2 mL, 298 mg, 2.612 mmol) was added at 0°C, and the mixture was warmed to 25°C and stirred for 1 h. After the reaction was completed, the reaction mixture was rotary evaporated to dryness to afford the crude compound 028h (60 mg).

[0231] MS m / z (ESI): 678.2 (M+1)+. Step 8 6-(4-((2,5,8,11,14-pentaoxahexadecan-16-yl)oxy)-2-(methylsulfonyl)pyrimidin-5-yl)-N-((2S,5S)-1-((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-5-isopropyl-2-methyl-1,4,7-trioxo-10,13-dioxa-3,6-diazapentadecan-15-yl)hex-5-ynamide (028) 028h (60 mg, 0.089 mmol), HATU (50 mg, 0.133 mmol), and N,N-diisopropylethylamine (34 mg, 0.266 mmol) were dissolved in N,N-dimethylformamide (1 mL). The compound 001c (51 mg, 0.089 mmol) was added at 0°C, and the reaction mixture was warmed to 25°C and stirred for 1 h. After the reaction was completed, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by preparative high-performance liquid chromatography (Shimadzu LH-40 Liquid Handler, Shimadzu LC-20AP Pump, Shimadzu SPD-20AP UV Detector; column: Xtimate C18,21.2*250 mm,5 um; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 20min gradient, gradient ratio: acetonitrile phase 20%-45%; flow rate: 20 mL / min) to afford the title compound 028 (28.2 mg, yield: 26%).

[0232] MS m / z (ESI): 1233.4 (M+1)+.

[0233] 1H NMR (400 MHz, DMSO-d6) 6 9.77 (s, 1H), 8.81 (s, 1H), 8.28 (d, 1H), 7.98 - 7.82 (m, 4H), 7.31 (s, 1H), 6.51 (s, 1H), 5.44 (s, 2H), 5.27 (s, 2H), 4.63 - 4.61 (m, 2H), 4.55 - 4.52 (m, 1H), 4.28 - 4.24 (m, 1H), 3.82 - 3.80 (m, 2H), 3.62 - 3.58 (m, 4H), 3.53 - 3.48 (m, 16H), 3.42 -3.39 (m, 10H), 3.23 - 3.15 (m, 8H), 3.00 - 2.96 (m, 2H), 2.26 (t, 2H), 2.06 - 1.97 (m, 3H), 1.91 - 1.84 (m, 2H), 1.82 - 1.75 (m, 2H), 1.40 (d, 3H), 0.91 - 0.84 (m, 9H). Example 15 6-(4-((2,5,8,11,14,17,20-heptaoxadocosan-22-yl)oxy)-2-(methylsulfonyl)pyrimidin-5-yl)-N-((2S,5S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H- benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-5-isopropyl-2-methyl-1,4,7-trioxo-10,13-dioxa-3,6-diazapentadecan-15-yl)hex-5-ynamide 029

[0234] Following the synthetic method in Example 14, the compound 029 was prepared according to the above synthetic route. The final product was purified by preparative high-performance liquid chromatography (Shimadzu LH-40 Liquid Handler, Shimadzu LC-20AP Pump, Shimadzu SPD-20AP UV Detectol; column: Xtimate C18,21.2*250 mm,5 um; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 20%-45%; flow rate: 20 mL / min) to afford the title compound 029 (10.5 mg, yield: 17%).

[0235] MS m / z (ESI): 661.4 (M / 2+1)+.

[0236] 1H NMR (400 MHz, DMSO-d6) 6 9.77 (s, 1H), 8.81 (s, 1H), 8.28 (d, 1H), 7.99 - 7.82 (m, 4H), 7.31 (s, 1H), 6.51 (s, 1H), 5.44 (s, 2H), 5.27 (s, 2H), 4.66 - 4.60 (m, 2H), 4.58 - 4.49 (m, 1H), 4.31 - 4.23 (m, 1H), 3.85 - 3.78 (m, 2H), 3.61-3.58 (m, 4H), 3.52-3.48 (m, 22H), 3.44 -3.38 (m, 7H), 3.23 - 3.15 (m, 7H), 2.98 (s, 2H),2.68 - 2.66 (m, 1H), 2.47 - 2.31 (m, 4H), 2.272.24 (m, 2H), 2.10 - 1.75 (m, 8H), 1.40 (d, 3H), 0.91 - 0.84 (m, 9H). Example 16 6-(4-((2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-28-yl)oxy)-2-(methylsulfonyl)pyrimidin-5-yl)-N-((2S,5S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-5-isopropyl-2-methyl-1,4,7-trioxo-10,13-dioxa-3,6-diazapentadecan-15-yl)hex-5-ynamide 030

[0237] Following the synthetic method in Example 14, the compound 030 was prepared according to the above synthetic route. The final product was purified by preparative high-performance liquid chromatography (Shimadzu LH-40 Liquid Handler, Shimadzu LC-20AP Pump, Shimadzu SPD-20AP UV Detector; column: Xtimate C18, 21.2*250 mm, 5 um; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 20%-45%; flow rate: 20 mL / min) to afford the title compound 030 (22.7 mg, 25%).

[0238] MS m / z (ESI): 1408.5 (M+1)+.

[0239] 1H NMR (400 MHz, DMSO-d6) 8 9.76 (s, 1H), 8.81 (s, 1H), 8.28 (d, 1H), 7.98 - 7.81 (m, 4H), 7.30 (s, 1H), 5.43 (s, 2H), 5.26 (s, 2H), 4.64 - 4.59 (m, 2H), 4.56 - 4.49 (m, 1H), 4.31 -4.22 (m, 1H), 3.83 - 3.78 (m, 2H), 3.61-3.58 (m, 5H), 3.52-3.50 (m, 4H), 3.49 - 3.37 (m, 30H), 3.27 - 3.14 (m, 8H), 2.97 (s, 2H), 2.53-2.50 (m, 2H), 2.48 - 2.33 (m, 4H), 2.27-2.23 (m, 2H), 2.07 - 1.74 (m, 8H), 1.40-1.38 (m, 3H), 0.90 - 0.84 (m, 9H). Example 17 (S)-N-(2-(((R)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H- benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-2-(2-(2-(10-(6-(4-methoxy-2- (methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)-1,7-dioxo-4,10-diazacyclododecan-4- yl)acetamido)-3-phenylpropanamide 040 Step 1 Tert-butyl 2-(1,7-dioxo-4,10-diazacyclododecan-4-yl)acetate 040b

[0240] The compound 040a (700 mg, 4.017 mmol), tert-butyl bromoacetate (705 mg, 3.616 mmol), and potassium carbonate (500 mg, 3.616 mmol) were dissolved in DMF (5 mL) and stirred at 25°C under nitrogen protection for 2 h. The reaction mixture was directly purified by reversed-phase chromatography (MeCN / H2O=1 / 1) to afford the title compound 040b (280 mg, 24.2%).

[0241] MS m / z (ESI): 288.9 (M+1) +. Step 2 Tert-butyl 2-(10-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynyl)-1,7-dioxo- 4,10-diazacyclododecan-4-yl)acetate 040c

[0242] The compound 040b (150 mg, 0.520 mmol), Int1 (206 mg, 0.520 mmol), and DIPEA (0.258 mL, 1.560 mmol) were dissolved in DMF (3 mL) and stirred at 25°C under N2 for 18 h. The reaction mixture was directly purified by reversed-phase chromatography (MeCN / H2O=1 / 1) to afford the title compound 040c (20 mg, 6.8%).

[0243] MS m / z (ESI): 569.2 (M+1) +. Step 3 2-(10-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynyl)-1,7-dioxo-4,10-diazacyclododecan-4-yl)acetic acid 040d

[0244] The compound 040c (20 mg, 0.035 mmol) was dissolved in a mixed solvent of DCM and TFA (1 mL, V / V = 1:1) and stirred at 25°C under nitrogen for 3 h. The reaction mixture was directly concentrated to afford the crude compound 040d (18 mg, 99.8%).

[0245] MS m / z (ESI): 512.7 (M+1)+. Step 4 (S)-N-(2-(((R)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[d]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-2-(2-(2-(10-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-ynyl)-1,7-dioxo-4,10-diazacyclododecan-4-yl)acetamido)-3-phenylpropanamide 040

[0246] The compound 040d (18 mg, 0.035 mmol), 019b (25 mg, 0.035 mmol), HATU (20 mg, 0.053 mmol), and DIPEA (0.017 mL, 0.105 mmol) were dissolved in DMF (2 mL) and stirred at 25°C for 1 h. After the reaction was completed, the reaction mixture was directly purified by preparative high-performance liquid chromatography (GX-281 Liquid Handler without pump, 4020 Syringe pump, 333-H3 Pump, 334-H3 Pump, 1741 U; column: Bonnasil-BS C18,20*250 mm,8um; mobile phase 1: water (containing 0.1% TFA); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 20%-50%; flow rate: 20 mL / min) to afford the compound 040 (5 mg, 12%).

[0247] MS m / z (ESI): 1215.3 (M+1)+.

[0248] 1H NMR (400 MHz, DMSO-d6) 8 9.69 (s, 1H), 8.90 (s, 1H), 8.78 (s, 1H), 8.52 (s, 1H), 8.27 - 8.19 (m, 2H), 7.99 - 7.86 (m, 3H), 7.31 (s, 1H), 7.28 - 7.25 (m, 3H), 7.20 - 7.17 (m, 1H), 6.51 (s, 1H), 5.43 (s, 2H), 5.27 (s, 2H), 4.59 (s, 1H), 4.19 - 4.00 (m, 7H), 3.90 - 3.49 (m, 18H), 3.25 - 2.93 (m, 8H), 2.84 - 2.82 (m, 1H), 2.67-2.66 (m, 1H), 2.33 - 2.31(m, 1H), 2.11 - 1.72 (m, 8H), 1.23 (s, 2H), 0.90-0.85 (m, 3H). Example 18 (S)-N-(2-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H- benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-2-oxoethyl)-2-(2-(2-(2-(16-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynyl)-1,4,10,13-tetraoxa-7,16- diazacyclooctadecan-7-yl)acetamido)-3-phenylpropanamide 041

[0249] The synthetic route of Example 17 was followed. The final product was purified by preparative high-performance liquid chromatography (Shimadzu LH-40 Liquid Handler, Shimadzu LC-20AP Pump, Shimadzu SPD-20AP UV Detector; column: Xtimate C18, 21.2*250 mm, 5 um; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; 20-min gradient, gradient ratio: acetonitrile phase 15%-40%; flow rate: 20 mL / min) to afford the title compound 041 (4.7 mg, yield: 4%).

[0250] MS m / z (ESI): 653.0 (M / 2+1)+.

[0251] 1H NMR (400 MHz, DMSO-d6) 6 9.63 (s, 1H), 8.80 (s, 1H), 8.51 - 8.47 (m, 1H), 8.22 -8.18 (m, 1H), 8.09 - 8.05 (m, 1H), 7.99 - 7.95 (m, 2H), 7.91 - 7.87 (m, 1H), 7.31 (s, 1H), 7.27 -7.22 (m, 4H), 7.21 - 7.14 (m, 1H), 6.51 (s, 1H), 5.44 (s, 2H), 5.27 (s, 2H), 4.59 - 4.55 (m, 1H), 4.07 (s, 3H), 4.03 - 4.00 (m, 2H), 3.78 - 3.70 (m, 3H), 3.61 - 3.56 (m, 6H), 3.56 - 3.47 (s, 22H), 3.17 - 3.11 (m, 4H), 3.11 - 3.07 (m, 4H), 3.03 - 2.96 (s, 2H), 2.88 - 2.82 (m, 2H), 2.07 - 2.00 (m, 2H), 1.91 - 1.84 (m, 2H), 1.80 - 1.77 (m, 2H), 0.89 (t, 3H). Example 19 N-(2-(((S)-1-(((S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-2-oxoethyl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)- N-(2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-28-yl)hex-5-ynamide 042

[0252] Following the synthetic route of Example 20, the starting material in step 1 was replaced with 042a. The final product was purified by preparative high performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge 5u C18 150 x 19 mm, 5 pm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 15-min gradient, gradient ratio: acetonitrile phase 35%-45%, flow rate: 20 mL / min) to give the compound 042 (16 mg, yield: 38%).

[0253] MS m / z (ESI): 1321.5 (M+1)+.

[0254] 1H NMR (400 MHz, DMSO-d6) 5 9.41 (d, 1H), 8.71 (s, 1H), 8.02 (d, 1H), 7.93 (d, 1H), 7.84 (d, 1H), 7.68 - 7.55(m, 1H), 7.32 (s, 1H), 6.16 (s, 1H), 5.40 (m, 2H), 5.22 (s, 2H), 4.56 (s, 1H), 4.30 - 4.20 (m, 1H), 3.54 - 3.47 (m, 34H), 3.47 - 3.38 (m, 5H), 3.33 (s,3H), 3.24 (s, 3H), 3.14 - 3.10 (m, 2H), 3.00 - 2.97 (m, 2H), 2.55 - 2.52 (m, 2H), 2.42 - 2.34 (m, 1H), 2.07 - 2.01 (m, 3H), 1.94 - 1.85 (m, 2H), 1.83 - 1.76(m, 2H), 1.44 - 1.38 (m, 3H), 1.25 (s, 2H), 0.92-0.87 (m, 9H). Example 20 N-(3-(((S)-1-(((S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H- benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropyl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)- N-(2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-28-yl)hex-5-ynamide 043 Step 1 Benzyl 29-((benzyloxy)carbonyl)-2,5,8,11,14,17,20,23,26-nonaoxa-29-azatriazetidin-32- oate 043b

[0255] Benzyl P-alanine p-toluenesulfonate 043a (300 mg, 0.85 mmol) was dissolved in acetonitrile (5 mL), and 28-bromo-2,5,8,11,14,17,20,23,26-nonaoxaoctacosane (419.5 mg, 0.85 mmol), potassium carbonate (352 mg, 2.55 mmol), and potassium iodide (141 mg, 0.85 mmol) were added. The mixture was stirred for reaction at 80°C for 16 h. After the reaction was completed, the mixture was cooled to room temperature, and benzyl chloroformate (290 mg, 1.7 mmol) was added and stirred for 30 min. After the reaction was completed, water was added, and the system was extracted with ethyl acetate. The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, and rotary evaporated to dryness to remove the solvent, thus giving a crude product. The crude product was purified by column chromatography with system B to afford 043b (400 mg, yield: 65%).

[0256] MS m / z (ESI): 724.3 (M+1)+. Step 2 2,5,8,11,14,17,20,23,26-nonaoxa-29-azatricyclopentadecan-32-oic acid 043c

[0257] The compound 043b (400 mg, 0.55 mmol) was dissolved in methanol (10 mL), and Pd / C (200 mg) was added. Then, the reaction was stirred at 25°C for 2 h. After the reaction was completed, the solvent was rotary evaporated to dryness to afford crude product 2,5,8,11,14,17,20,23,26-nonaoxa-29-azatricyclopentadecan-32-oic acid 043c (260 mg). The crude product was directly used in the next reaction step without purification.

[0258] MS m / z (ESI): 500.3 (M+1)+. Step 3 29-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamoyl)-2,5,8,11,14,17,20,23,26-nonaoxa-29-azadotriacontan-32-oic acid 043d

[0259] The crude product 2,5,8,11,14,17,20,23,26-nonaoxa-29-azatricyclopentadecan-32-oic acid 043c (130 mg, 0.26 mmol) was dissolved in DMF (2 mL). The compound Int1 (103 mg, 0.26 mmol) and DIPEA (50 mg, 0.39 mmol) were added. The mixture was stirred for reaction at room temperature for 1 h. After the reaction was completed, the reaction mixture was purified by preparative high-performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: XBridge 5u C18 100 x 19 mm, 5 pm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 10-min gradient, gradient ratio: acetonitrile phase 27%-37%, flow rate: 20 mL / min) to afford 29-(6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamoyl)-2,5,8,11,14,17,20,23,26-nonaoxa-29-azadotriacontan-32-oic acid 043d (71 mg, yield: 35%).

[0260] MS m / z (ESI):780.4 (M+1)+. Step 4 N-(3-(((S)-1-(((S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropyl)-6-(4-methoxy-2- (methylsulfonyl)pyrimidin-5-yl)-N-(2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-28-yl)hex-5-ynamide 043

[0261] The compound 043d (25 mg, 0.032 mmol) was dissolved in N,N-dimethylformamide (1 mL), and the compound 001c (25 mg, 0.044 mmol), HATU (18 mg, 0.048 mmol), and DIPEA (9 mg, 0.64 mmol) were added. The mixture was stirred for reaction at room temperature for 1 h. After the reaction was completed, the reaction mixture was purified by preparative high performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge 5u C18 150 x 19 mm, 5pm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 10-min gradient, gradient ratio: acetonitrile phase 38%-48%, flow rate: 20 mL / min) to give the compound 043 (8.7 mg, yield: 20%).

[0262] MS m / z (ESI): 668.5 (M / 2+1)+.

[0263] 1H NMR (400 MHz, DMSO-d6) 8 9.75 (d, 1H), 8.80 (d, 1H), 8.29 (dd, 1H), 8.11 - 7.98 (m, 1H), 7.97 - 7.93 (m, 1H), 7.82 (d, 1H), 7.30 (s, 1H), 6.50 (s, 1H), 5.43 (s, 2H), 5.26 (s, 2H), 4.57 - 4.48 (m,1H), 4.34 - 4.18 (m, 1H), 4.07 (s, 3H), 3.51 - 3.48 (m, 30H), 3.47 - 3.45 (m, 5H), 3.43 - 3.41 (m, 3H), 3.39 (s, 3H), 3.23 (s, 3H), 3.18 - 3.13 (m, 2H), 3.01 - 2.94 (m, 2H), 2.06 - 1.95 (m, 4H), 1.92 - 1.83 (m, 2H), 1.82 - 1.74 (m, 2H), 1.42 - 1.36 (m , 3H), 1.28 - 1.21 (m, 2H), 0.91 - 0.81 (m, 12H). Example 21 N-(5-(((S)-1-(((S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-5-oxopentyl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)- N-(2,5,8,11,14,17,20,23,26-nonaoxaoctacosan-28-yl)hex-5-ynamide 044 BocHN 044a

[0264] Following the synthetic route of Example 20, the starting material in step 1 was replaced with 044a. The final product was purified by preparative high performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge 5um C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 15-min gradient, gradient ratio: acetonitrile phase 35%-45%, flow rate: 20 mL / min) to give the compound 044 (20 mg, yield: 35%).

[0265] MS m / z (ESI): 1364.5 (M+1)+.

[0266] 1H NMR (400 MHz, CD3OD) 59.77 (s, 1H), 8.80 (s, 1H), 8.26 (t, 1H), 8.02 - 7.93 (m, 1H), 7.91 - 7.80 (m, 2H), 7.31 (s, 1H), 6.52 (s, 1H), 5.44 (s, 2H), 5.27 (s, 2H), 4.57 - 4.49 (m, 1H), 4.27 - 4.20 (m, 1H), 4.11 - 4.04 (m, 3H), 3.53 - 3.46 (m, 32H), 3.44 - 3.41 (m, 10H), 3.25 - 3.23 (m, 3H), 3.20 - 3.14 (m, 2H), 3.01 - 2.95 (m, 2H), 2.27 - 2.14 (m, 2H), 2.12 - 1.93 (m, 4H), 1.91 - 1.74 (m, 4H), 1.57 - 1.32 (m, 8H), 1.24 (s, 1H), 0.92 - 0.84 (m, 9H). Example 22 N-(4-(((S)-1-(((S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-4-oxobutyl)-6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)- N-(2,5,8,11,14,17,20,23,26,29,32,35,38-tridecaoxatetracontan-40-yl)hex-5-ynamide 045 Step 1 2,5,8,11,14,17,20,23,26,29,32,35,38-tridecaoxatetracontan-40-yl-4-methylbenzenesulfonate 045b

[0267] 2,5,8,11,14,17,20,23,26,29,32,35,38-tridecaoxatetracontan-40-ol 045a (1 g, 1.7 mmol) was dissolved in dichloromethane (20 mL), and p-toluenesulfonyl chloride (0.65 g, 3.4 mmol), triethylamine (340 mg, 3.4 mmol), and 4-dimethylaminopyridine (1.04 g, 8.5 mmol) were added sequentially. The mixture was stirred at room temperature for 16 h. After the reaction was completed, the solvent was rotary evaporated to dryness, the mixture was concentrated under reduced pressure to afford the crude product. The crude product was purified by silica gel column chromatography to afford the compound 045b (1.2 g, yield: 94%).

[0268] MS m / z(ESI): 759.3 (M+1)+. Step 2 Benzyl 2,5,8,11,14,17,20,23,26,29,35,38-tridecaoxa-41-azapentatetracontan-45-oate 045d

[0269] Benzyl 4-aminobutanoate 045c (112 mg, 0.58 mmol) was dissolved in acetonitrile (10 mL). Potassium carbonate (146 mg, 1.05 mmol) and 2,5,8,11,14,17,20,23,26,29,32,35,38-tridecaoxatetracontan-40-yl-4-methylbenzenesulfonate 045b (400 mg, 0.53 mmol) were added. The reaction mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was used directly in the next step without purification to afford the crude product 045d (411 mg).

[0270] MS m / z(ESI): 780.4(M+1)+. Step 3 Benzyl 41-((benzyloxy)carbonyl)-2,5,8,11,14,17,20,23,26,29,32,35,38-tridecaoxa-41-azapentatetracontan-45-oate 045e

[0271] The compound 045d (411 mg, 0.53 mmol) was dissolved in acetonitrile (10 mL). Benzyl chloroformate (108 mg, 0.64 mmol) was added, and the mixture was stirred at room temperature for 1 h. After the reaction was completed, the solvent was rotary evaporated to dryness, the mixture was concentrated under reduced pressure to afford the crude compound 045e (180 mg, yield: 37%).

[0272] MS m / z(ESI): 931.4(M+18)+. Step 4 2,5,8,11,14,17,20,23,26,29,32,35,38-tridecaoxa-41-azapentatetracontan-45-oic acid 045f

[0273] The compound 045e (180 mg, 0.2 mmol) was dissolved in methanol (10 mL). Palladium on carbon (35 mg, 0.2 mmol) was added. The reaction mixture was stirred under a hydrogen atmosphere at room temperature for 1 h. After the reaction was completed, the mixture was filtered directly to afford the compound 045f (120 mg, 88%).

[0274] MS m / z(ESI): 690.4 (M+1)+. Step 5 41-(6-(4-methoxy-2-(methanesulfonyl)pyrimidin-5-yl)hex-5-ynoyl)-2,5,8,11,14,17,20,23,26,29,32,35,38-tridecaoxa-41-azapentatetracontan-45-oic acid 045g

[0275] The compound 045f (120 mg, 0.3 mmol) was dissolved in N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (78 mg, 0.6 mmol) and 2,5-dioxopyrrolidin-1-yl 6-(4-methoxy-2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynoate Int-1 (230 mg, 0.33 mmol) were added thereto. The reaction mixture was stirred at room temperature for 1 h. Upon reaction completion, the crude product was purified via preparative high performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge 5u C18 150 x 19 mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; gradient ratio: acetonitrile phase 30%-40%, flow rate: 20 mL / min) to afford the compound 045g (60 mg, yield: 20%).

[0276] MS m / z(ESI): 970.3(M+1)+. Step 6 N-(4-(((S)-1-(((S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-4-oxobutyl)-6-(4-methoxy-2- (methylsulfonyl)pyrimidin-5-yl)-N-(2,5,8,11,14,17,20,23,26,29,32,35,38-tridecaoxatetracontan-40-yl)hex-5-ynamide 045

[0277] The compound 045g (50 mg, 0.05 mmol) was dissolved in N,N-dimethylformamide (10 mL). The compound 001c (32.5 mg, 0.06 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N',N'- tetramethyluronium hexafluorophosphate (30 mg, 0.08 mmol), and N,N-diisopropylethylamine (14 mg, 0.1 mmol) were added thereto. The mixture was stirred at room temperature for 2 h. After the reaction was completed, the solvent was rotary evaporated to dryness to afford the crude product. The crude product was purified by preparative high performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge 5u C18 150 x 19 mm; mobile phase 1: water (containing 0.1% formic acid); mobile phase 2: acetonitrile; gradient ratio: acetonitrile phase 37%-47%, flow rate: 20 mL / min) to give the compound 045 (25.9 mg, yield: 33%).

[0278] MS m / z(ESI): 763.8(M / 2+1)+.

[0279] 1H NMR (400 MHz, CDCI3) 8 8.97 (s, 1H), 8.56 (s, 1H), 8.02 (s, 1H), 7.93 - 7.81 (m, 2H), 7.61 (s, 1H), 6.63 (s, 1H), 5.69 (d, 1H), 5.30 - 5.23(m, 1H), 5.14 - 5.01 (m, 2H), 4.84 -4.76 (m, 1H), 4.28 (s, 1H), 4.17 (s, 3H), 3.66 - 3.62 (m, 48H), 3.39 - 3.37 (m, 4H), 3.34 - 3.33 (m, 3H), 3.11 - 2.94 (m, 5H), 2.66 - 2.58 (m, 4H), 2.40 - 2.29 (m, 2H), 2.14 (s, 4H), 1.98 - 1.87 (m, 6H), 1.70 (s, 1H), 1.57 - 1.50 (m, 3H), 1.08 - 0.94 (m, 12H). Example 23 N-((S)-1-(((S)-1-(((S)-9-ethyl-9-hydroxy-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-1-(6-(2-(methylsulfonyl)pyrimidin-5-yl)hex-5-ynamido)-3,6,9,12,15,18,21,24-octaoxaheptacosan-27-amide

[0280] The compound 057 was obtained using the same synthesis method as in the examples.

[0281] MS m / z(ESI): 624.3 (M / 2+1)+.

[0282] 1H NMR (400 MHz, CD3OD) 5 8.95 (s, 1H), 8.94 (s, 1H), 7.96 (d, 1H), 7.81 (d, 1H), 7.65 (s, 1H), 5.58 (d, 1H), 5.38 (d, 1H), 5.33 (t, 1H), 5.25 (s, 2H), 4.64-4.57 (m, 2H), 4.26-4.22 (m, 1H), 3.73 (t, 3H), 3.62-3.58 (m, 30H), 3.54-3.50 (m, 2H), 3.36-3.33 (m, 2H), 3.22-3.17 (m, 2H), 3.09-3.01 (m, 2H), 2.60-2.53 (m, 2H), 2.38 (t, 2H), 2.21-2.10 (m, 2H), 2.04-1.99 (m, 3H), 1.98-1.88 (m, 4H), 1.54 (d, 3H), 1.04-0.97 (m, 9H).

[0283] Following the synthesis method described in patent WO2020200880A1, the following compound 058 was synthesized: Preparation of antibody-drug conjugates Example A-1 Preparation of ADC-1

[0284] Trastuzumab (16 mg) was placed in a 50 mL centrifuge tube. 50 mM PBS, PH7.97, was added to adjust the antibody concentration to 5 mg / mL. 0.2 M EDTA was added to achieve a final EDTA concentration of 2 mM, followed by the addition of 15 molar equivalents of TCEP. The mixture was incubated at 37°C for 2 h with continuous mixing. Under ice bath conditions, a solution of compound 001 (formulated in DMA) was added at a final drug-to-antibody molar ratio of 15:1. DMA was supplemented to account for 10% of the total reaction volume. After vortex mixing, the reaction was incubated at 22°C for 3 h, followed by reaction at 4°C for 18 h with continuous mixing. The reaction mixture was purified using a Zeba desalting column. The sample was concentrated using Amicon and exchanged to a 30 mM His / HAc buffer system with a pH of 5.5, resulting in ADC-1 (14 mg). The average DAR value (P) of the ADC was determined to be 8 by HIC analysis.

[0285] Following the procedure of Example A-1 and replacing the compound 001 with compounds from other examples of the present application, the following antibody-drug conjugates were prepared. Their structures and DAR values (P) are shown in the table below. ADC Structure ADC mass (mg) DAR value (P) ADC-2 Trastuzumab Ho'X / y g                      S Y h j nNT w’ l / , / / A, / , ,o, / / / , ,o, / , / , ..o., / / / , ,o, / / / , A, A, „NjsiX AL, / L, A-o A H                            H iX H U _                                                ADC-2                                                   J 12 8 ADC-3 Tratuzumab. HO'. L ^jy 0                                               0X"h 0 0Me^' / ^'''XAA''' / 'CAA'A^'' / 'CAA'A^^^                                                    ° H                                                                                                                            H II i H 1 I ADC-3 0 13 8 ADC-5 Trastuzumab 'A ° CN                                                                                             HO'.l—# I                                                 yV ° N<X    g                          jYh L fXNTX-X H                                             H     i ” XJ _                                                ADC-5                                                   J 10 8 ADC-8 Trastuzumab HoXX N                                                    Y\'X NgX, ?                            ? A h s / / A / / o / ' / / o, / / q / ^^—° OH                      H H i H |^J _                                                ADC-8                                                   J 11 8 ADC- 11 Trastuzumab _,N. NyX H        h 2 1 h V OMe    ''' -'AA ''°^                            ,0 j    5 a H o U X / X r                       N v / ~\ r°                              Ai / 0 HO<A\ / o ADC-11 3 10 8 ADC- 12 Trastuzumab. 1 < X Pa ( : > )         oV □ / \ o o            \ zi » >     > v O           IZ (         X— ) ° \ O              ZI Y \.XA i2 l O r°         i 3 12 8 ADC- 42 Trastuzumab- NvX,      || H u I H XP OMe        N - Ar N   N Xy N Y^yY'Ai / P o' VA UpA r°                    >a0 L                           HOYA ~ o'YA' / Ar                     A 0 , / / / ,0, / ^ / / ,0.    ADC-42 0               0    ■' p 13 8 ADC- 43 A H 9 I H n lMe                                     f o J a H S U X / M A J                N tA Trastuzumab' N                                                            ( o ADC-43                           / / 0 p 14 8 ADC- 57 Trastuzumab. HO'.L^ n                                                            aa2 / ° °                                       ° Y h °           -4 / aj.n^»^o^«^o^"^o^»^an1Ln«anaan h                       h A h A -                                               ADC-57 p 49 8 ADC- 58 Trastuzumab _                                              ADC-58 0 16 7.7

[0286] The results show that the cytotoxic drug-linker compounds of the present invention can be smoothly conjugated with antibodies to give antibody-drug conjugates. Biological Evaluation Test Example 1 ADC biological activity assay 1. Test purpose

[0287] The purpose of this experiment is to detect the inhibitory activity of ADC compounds on the proliferation of HER2-expressing NCI-N87 cells, SK-BR-3 cells and HER2-negative MDA-MB-468 cells in vitro. Cells were treated with different concentrations of the compound in vitro. After 5 days of culture, cell proliferation was detected using CTG CellTiter-Glo® Luminescent Cell Viability Assay, and the in vitro activity of the compound was evaluated based on the IC50 value. 2. Test method (1) On the first day, tumor cells were plated in a 96-well plate, with 5000 cells / 100 pL culture medium in each well, and 100 pL of DPBS was seeded in each empty well at the edge. The cells were incubated overnight in a 37°C incubator. (2) On the second day, the old culture medium (50 pL / well) was removed by pipetting; ADC of different concentration gradients was added, with the starting concentration of ADC being 200 nM, and 5-fold dilutions for 9 concentrations. The dosing volume was 50 pL / well. (3) On the sixth day, CellTiter-Glo Buffer and CellTiter-Glo Substrate reagents were thawed at 4°C. Before use, 10 ml of buffer was taken by pipetting and added to the substrate, the system was mixed well, and equilibrated to room temperature. (4) On the seventh day, the 96-well plate was equilibrated at room temperature for 30 min and 100 pL of Cell-Titer-Glo was added to each well. After shaking at room temperature in the dark for 5 min, the system was incubated for 10 min, 100 pL of the liquid in the well was transferred to the blank plate, and then detected for chemiluminescence with a microplate reader. 3. Data analysis

[0288] The data were processed and analyzed using Microsoft Excel and Graphpad Prism 5. The inhibitory activity of the ADC compounds against the proliferation of NCI-N87 cells, SK-BR-3 cells, and MDA-MB-468 cells in vitro were tested, and the results are shown in Table 1 below. Table 1 Compound IC50 (nM) NCI-N87 SK-BR-3 MDA-MB-468 DS8201 0.36 0.06 >50 ADC-1 0.31 0.03 >50 ADC-2 0.33 0.03 >50 ADC-3 0.28 0.02 >50 ADC-5 0.26 0.02 >50

[0289] Conclusion: The antibody-drug conjugates targeting HER2 of the present invention have obvious proliferation inhibitory activity against HER2-positive cells SK-BR-3 and NCI-N87; meanwhile, they have weak proliferation inhibitory activity against HER2-negative cells MDA-MB-468; therefore, they have good selectivity. Test Example 2 ADC plasma stability experiment

[0290] The mice used in this experiment were CD-1 mice, the rats used were SD rats, and the monkeys used were cynomolgus monkeys. (1) Free toxin release assay and results

[0291] The DS8201 sample, ADC-2, ADC-12, and ADC-58 were individually added to the above sterile mouse plasma, sterile rat plasma, sterile human plasma, and sterile monkey plasma at a final concentration of 200 pgmL. The mixtures were incubated in a 37°C cell incubator. The day of incubation initiation was designated as Day 0. Samples were subsequently taken out on Day 1, Day 4, Day 7, Day 14, and Day 21 for detection of free toxin content.

[0292] The free toxin release rates are shown in Fig. 1 to Fig. 3 and Fig. 14. The results indicate that ADC-2 and ADC-12 exhibit excellent stability in mouse, rat, human, and monkey plasma, with the maximum free toxin release rate not exceeding 0.2%. Furthermore, their stability was significantly superior to that of the reference DS8201 and ADC-58. (2) ADC DAR value test and results

[0293] The DS8201 sample, ADC-2, ADC-12, and ADC-58 were added to the above-mentioned sterile human plasma at a final concentration of 200 ug / mL and incubated in a 37°C cell culture incubator. The day when incubation started was recorded as day 0, and then samples were taken out on day 1, day 4, day 7, day 14, and day 21, respectively, to detect changes in DAR values.

[0294] Table 2 shows the experimental results of ADC DAR value change. According to the results, the change of DAR value of the conjugate formed by the small molecule linker of the present invention in human plasma is significantly smaller than that of DS8201 and ADC-58, showing better plasma stability, which further confirms the stability of the small molecule linker of the present invention. Table 2: ADC plasma stability (DAR value change) Compound DAR value (0 Day) DAR value (1 Day) DAR value (4 Day) DAR value (7 Day) DAR value (14 Day) DAR value (21 Day) DS8201 7.62 5.96 4.75 4.31 3.74 3.58 ADC-2 7.73 7.73 7.76 7.73 7.71 7.73 ADC-12 7.85 7.79 7.78 7.77 7.76 7.74 ADC-58 7.58 6.52 6.38 6.34 6.25 6.27 Test Example 3: Efficacy evaluation in NCI-N87 tumor-bearing mice 3.1 Test objective

[0295] To evaluate the efficacy of the ADCs in the present application by investigating whether tumor growth is inhibited, delayed or cured in Balb / c nude mice as test animals. 3.2 Experimental steps 3.2.1 Test drugs

[0296] Blank control / or Vehicle: PBS Reference ADC (DS8201): 3 mg / kg ADC-2: 3 mg / kg ADC-12: 3 mg / kg ADC-43: 1 mg / kg 3.2.2 Preparation method: All of them were diluted with PBS. 3.2.3 Experimental method

[0297] NCI-N87 cells were seeded subcutaneously in the right rib of mice. After the tumors grew for 7 days, the animals were randomly divided into groups, 6 animals per group, for a total of 5 groups (4 treatment groups + 1 blank control group). The drug was administered once by tail vein injection. Tumor volume (diameter) and body weight were measured twice a week for four weeks, and the data were recorded. Data statistics were calculated using Excel 2023 statistical software: the mean value was calculated as avg; the SD value was calculated as STDEV; the SEM value was calculated as STDEV / SQRT; and the P value of the difference between groups was calculated as TTEST.

[0298] Tumor volume calculation formula: V = 0.5 a x b2 where a and b represent the long diameter and short diameter of the tumor, respectively.

[0299] The anti-tumor efficacy of the compound was evaluated by TGI (%), and the tumor growth inhibition rate was calculated using the following formula: TGI (%) = [1 - (Ti - T0) / (Ci - C0)] x 100%, where Ti is the average tumor volume of a treatment group on a given day, T0 is the average tumor volume of the same treatment group at the start of drug administration; Ci is the average tumor volume of the vehicle control group on a given day (the same day as Ti), and C0 is the average tumor volume of the vehicle control group at the start of drug administration. 3.3 Experimental results and conclusions

[0300] The schematic diagram of efficacy evaluation in NCI-N87 tumor-bearing mice is shown in Fig. 4. The in vivo tumor growth inhibition (TGI) effects of the test drugs in the treatment groups on the NCI-N87 xenograft model are shown in Table 3. The results show that the ADC molecules of the present application can significantly reduce the tumor volume and have better tumor inhibition effects than the reference ADC (DS8201). Table 3: In vivo tumor growth inhibition effects of ADCs on NCI-N87 transplant model Group Test drug TGI (%) Group 1 / (Vehicle) / Group 2 Reference ADC (DS8201) (3mpk) 105 Group 3 ADC-2 (3mpk) 115 Group 4 ADC-12 (3mpk) 110 Group 5 ADC-43 (1mpk) 115 Test Example 4: Efficacy evaluation in JIMT-1 tumor-bearing mice 4.1 Test objective

[0301] To evaluate the efficacy of the ADCs in the present application by investigating whether tumor growth is inhibited, delayed or cured in SCID Beige mice as test animals. 4.2 Experimental steps 4.2.1 Test drugs

[0302] Blank control / or Vehicle: PBS Reference ADC (DS8201): 1 mg / kg ADC-2: 1 mg / kg ADC-5: 1 mg / kg ADC-8: 1 mg / kg ADC-11: 1 mg / kg ADC-12: 1 mg / kg 4.2.2 Preparation method: All of them were diluted with PBS. 4.2.3 Experimental method

[0303] JIMT-1 cells were seeded subcutaneously in the right rib of mice. After the tumors grew for 8 days, the animals were randomly divided into groups, 6 animals per group, for a total of 7 groups (6 treatment groups + 1 blank control group). The drug was administered once by tail vein injection. Tumor volume (diameter) and body weight were measured twice a week for four weeks, and the data were recorded. Data statistics were performed using Excel 2023 statistical software: the mean value was calculated as avg; the SD value was calculated as STDEV; the SEM value was calculated as STDEV / SQRT; and the P value of the difference between groups was calculated as TTEST.

[0304] Tumor volume calculation formula: V = 0.5 a x b2 where a and b represent the long diameter and short diameter of the tumor, respectively.

[0305] The anti-tumor efficacy of the compound was evaluated by TGI (%), and the tumor growth inhibition rate was calculated using the following formula: TGI (%) = [1 - (Ti - T0) / (Ci - C0)] x 100%, where Ti is the average tumor volume of a treatment group on a given day, T0 is the average tumor volume of the same treatment group at the start of drug administration; Ci is the average tumor volume of the vehicle control group on a given day (the same day as Ti), and C0 is the average tumor volume of the vehicle control group at the start of drug administration. 4.3 Experimental results and conclusions

[0306] The in vivo tumor growth inhibition effects of the treatment groups on the JIMT-1 xenograft model are shown in Table 4, and the schematic diagram of efficacy evaluation in JIMT-1 tumor-bearing mice is shown in Fig. 5. The results show that the ADC molecules of the present application can significantly reduce the tumor volume and have better tumor inhibition effects than the reference ADC (DS8201). Table 4: In vivo tumor growth inhibition effects of ADCs on JIMT-1 transplant model Group Test drug TGI (%) Group 1 / (Vehicle) / Group 2 Reference ADC (DS8201) (1mpk) 62 Group 3 ADC-2 (1mpk) 111 Group 4 ADC-5 (1mpk) 114 Group 5 ADC-8 (1mpk) 103 Group 6 ADC-11 (1mpk) 112 Group 7 ADC-12 (1mpk) 117 Test Example 5: Efficacy evaluation in RT11284 tumor-bearing mice 5.1 Test objective

[0307] To evaluate the efficacy of the ADCs in the present application by investigating whether tumor growth is inhibited, delayed or cured in BALB / c Nude mice as test animals. 5.2 Experimental steps 5.2.1 Test drugs

[0308] Blank control / or Vehicle: PBS Reference ADC (DS8201): 3 mg / kg Reference ADC (DS8201): 10 mg / kg ADC-2: 3 mg / kg ADC-2: 10 mg / kg ADC-12: 3 mg / kg ADC-42: 3 mg / kg ADC-43: 3 mg / kg 5.2.2 Preparation method: All of them were diluted with PBS. 5.2.3 Experimental method

[0309] RT11284 cells were seeded subcutaneously in the right rib of mice. After the tumors grew for 10 days, the animals were randomly divided into groups, 6 animals per group, for a total of 8 groups (7 treatment groups + 1 blank control group). The drug was administered once by tail vein injection. Tumor volume (diameter) and body weight were measured twice a week for four weeks, and the data were recorded. Data statistics were performed using Excel 2023 statistical software: the mean value was calculated as avg; the SD value was calculated as STDEV; the SEM value was calculated as STDEV / SQRT; and the P value of the difference between groups was calculated as TTEST.

[0310] Tumor volume calculation formula: V = 0.5 a x b2 where a and b represent the long diameter and short diameter of the tumor, respectively.

[0311] The anti-tumor efficacy of the compound was evaluated by TGI (%), and the tumor growth inhibition rate was calculated using the following formula: TGI (%) = [1 - (Ti - T0) / (Ci - C0)] x 100%, where Ti is the average tumor volume of a treatment group on a given day, T0 is the average tumor volume of the same treatment group at the start of drug administration; Ci is the average tumor volume of the vehicle control group on a given day (the same day as Ti), and C0 is the average tumor volume of the vehicle control group at the start of drug administration. 5.3 Experimental results and conclusions

[0312] The schematic diagram of efficacy evaluation in RT11284 tumor-bearing mice is shown in Fig. 6. The in vivo tumor growth inhibition effects of the test drugs in the treatment groups on the RT11284 xenograft model are shown in Table 5. The results show that the ADC molecules of the present application can significantly reduce the tumor volume and have better tumor inhibition effects than the reference ADC (DS8201). Table 5: In vivo tumor growth inhibition effects of ADCs on RT11284 transplant model Group Test drug TGI (%) Group 1 / (Vehicle) / Group 2 Reference ADC (DS8201) (3mpk) 56 Group 3 Reference ADC (DS8201) (10mpk) 79 Group 4 ADC-2 (3mpk) 99 Group 5 ADC-2 (10mpk) 117 Group 6 ADC-12 (3mpk) 94 Group 7 ADC-42 (3mpk) 94 Group 8 ADC-43 (3mpk) 89 Test Example 6: Efficacy evaluation in HCT116 tumor-bearing mice 6.1 Test objective

[0313] To evaluate the efficacy of the ADCs in the present application by investigating whether tumor growth is inhibited, delayed or cured in BALB / c Nude mice as test animals. 6.2 Experimental steps 6.2.1 Test drugs

[0314] Blank control / or Vehicle: PBS Reference ADC (DS8201): 3 mg / kg Reference ADC (DS8201): 10 mg / kg ADC-43: 3 mg / kg ADC-43: 10 mg / kg ADC-2: 3 mg / kg ADC-2: 10 mg / kg 6.2.2 Preparation method: All of them were diluted with PBS. 6.2.3 Experimental method

[0315] HCT116 cells were seeded subcutaneously in the right rib of mice. After the tumors grew for 14 days, the animals were randomly divided into groups, 6 animals per group, for a total of 7 groups (6 treatment groups + 1 blank control group). The drug was administered once by tail vein injection. Tumor volume (diameter) and body weight were measured twice a week for four weeks, and the data were recorded. Data statistics were performed using Excel 2023 statistical software: the mean value was calculated as avg; the SD value was calculated as STDEV; the SEM value was calculated as STDEV / SQRT; and the P value of the difference between groups was calculated as TTEST.

[0316] Tumor volume calculation formula: V = 0.5 a x b2 where a and b represent the long diameter and short diameter of the tumor, respectively.

[0317] The anti-tumor efficacy of the compound was evaluated by TGI (%), and the tumor growth inhibition rate was calculated using the following formula: TGI (%) = [1 - (Ti - T0) / (Ci - C0)] x 100%, where Ti is the average tumor volume of a treatment group on a given day, T0 is the average tumor volume of the same treatment group at the start of drug administration; Ci is the average tumor volume of the vehicle control group on a given day (the same day as Ti), and C0 is the average tumor volume of the vehicle control group at the start of drug administration. 6.3 Experimental results and conclusions

[0318] The schematic diagram of efficacy evaluation in HCT116 tumor-bearing mice is shown in Fig. 7. The in vivo tumor growth inhibition effects of the test drugs in the treatment groups on the HCT116 xenograft model are shown in Table 6. The results show that the ADC molecules of the present application can significantly reduce the tumor volume and have better tumor inhibition effects than the reference ADC (DS8201). Table 6: In vivo tumor growth inhibition effects of ADCs on HCT116 transplant model Group Test drug TGI (%) Group 1 / (Vehicle) / Group 2 Reference ADC (DS8201) (3mpk) 26 Group 3 Reference ADC (DS8201) (10mpk) 55 Group 4 ADC-43 (3mpk) 62 Group 5 ADC-43 (10mpk) 94 Group 6 ADC-2 (3mpk) 68 Group 7 ADC-2 (10mpk) 91 Test Example 7: Efficacy evaluation in Capan-1 tumor-bearing mice 7.1 Test objective

[0319] To evaluate the efficacy of the ADCs in the present application by investigating whether tumor growth is inhibited, delayed or cured in SCID Beige mice as test animals. 7.2 Experimental steps 7.2.1 Test drugs

[0320] Blank control / or Vehicle: PBS ADC (DS8201): 3 mg / kg ADC (DS8201): 10 mg / kg ADC-2: 3 mg / kg ADC-2: 10 mg / kg ADC-12: 3 mg / kg ADC-12: 10 mg / kg ADC-42: 3 mg / kg ADC-43: 3 mg / kg 7.2.2 Preparation method: All of them were diluted with PBS. 7.2.3 Experimental method

[0321] Capan-1 cells were seeded subcutaneously in the right rib of mice. After the tumors grew for 8 days, the animals were randomly divided into groups, 6 animals per group, for a total of 9 groups (8 treatment groups + 1 blank control group). The drug was administered once by tail vein injection. Tumor volume (diameter) and body weight were measured twice a week for four weeks, and the data were recorded. Data statistics were performed using Excel 2023 statistical software: the mean value was calculated as avg; the SD value was calculated as STDEV; the SEM value was calculated as STDEV / SQRT; and the P value of the difference between groups was calculated as TTEST.

[0322] Tumor volume calculation formula: V = 0.5 a x b2 where a and b represent the long diameter and short diameter of the tumor, respectively.

[0323] The anti-tumor efficacy of the compound was evaluated by TGI (%), and the tumor growth inhibition rate was calculated using the following formula: TGI (%) = [1 - (Ti - T0) / (Ci - C0)] x 100%, where Ti is the average tumor volume of a treatment group on a given day, T0 is the average tumor volume of the same treatment group at the start of drug administration; Ci is the average tumor volume of the vehicle control group on a given day (the same day as Ti), and C0 is the average tumor volume of the vehicle control group at the start of drug administration. 7.3 Experimental results and conclusions

[0324] The schematic diagram of efficacy evaluation in Capan-1 tumor-bearing mice is shown in Fig. 8. The in vivo tumor growth inhibition (TGI) effects of the test drugs in the treatment groups on the Capan-1 xenograft model are shown in Table 7. The results show that the ADC molecules of the present application can significantly reduce the tumor volume and have better tumor inhibition effects than the reference ADC (DS8201). Table 7: In vivo tumor growth inhibition effects of ADCs on Capan-1 transplant model Group Test drug TGI (%) Group 1 / (Vehicle) / Group 2 Reference ADC (DS8201) (3mpk) 83 Group 3 Reference ADC (DS8201) (10mpk) 104 Group 4 ADC-2 (3mpk) 104 Group 5 ADC-2 (10mpk) 105 Group 6 ADC-12 (3mpk) 104 Group 7 ADC-12 (10mpk) 105 Group 8 ADC-42 (3mpk) 104 Group 9 ADC-43 (3mpk) 104 Test Example 8: Efficacy evaluation in NCI-N87- Enhertu-resistant cell tumor-bearing mice 8.1 Test objective

[0325] To evaluate the efficacy of the ADCs in the present application by investigating whether tumor growth is inhibited, delayed or cured in NOD SCID mice as test animals. 8.2 Experimental steps 8.2.1 Test drugs

[0326] Blank control / or Vehicle: PBS Reference ADC (DS8201): 3 mg / kg Reference ADC (DS8201): 10 mg / kg ADC-2: 3 mg / kg ADC-2: 10 mg / kg ADC-43: 3 mg / kg 8.2.2 Preparation method: All of them were diluted with PBS. 8.2.3 Experimental method

[0327] NCI-N87-Enhertu-resistant cells were seeded subcutaneously in the right rib of mice. After the tumors grew for 12 days, the animals were randomly divided into groups, 6 animals per group, for a total of 6 groups (5 treatment groups + 1 blank control group). The drug was administered once by tail vein injection. Tumor volume (diameter) and body weight were measured twice a week for four weeks, and the data were recorded. Data statistics were performed using Excel 2023 statistical software: the mean value was calculated as avg; the SD value was calculated as STDEV; the SEM value was calculated as STDEV / SQRT; and the P value of the difference between groups was calculated as TTEST.

[0328] Tumor volume calculation formula: V = 0.5 a x b2 where a and b represent the long diameter and short diameter of the tumor, respectively.

[0329] The anti-tumor efficacy of the compound was evaluated by TGI (%), and the tumor growth inhibition rate was calculated using the following formula: TGI (%) = [1 - (Ti - T0) / (Ci - C0)] x 100%, where Ti is the average tumor volume of a treatment group on a given day, T0 is the average tumor volume of the same treatment group at the start of drug administration; Ci is the average tumor volume of the vehicle control group on a given day (the same day as Ti), and C0 is the average tumor volume of the vehicle control group at the start of drug administration. 8.3 Experimental results and conclusions

[0330] The schematic diagram of efficacy evaluation in NCI-N87- Enhertu-resistant cell tumorbearing mice is shown in Fig. 9. The in vivo tumor growth inhibition (TGI) effects of the test drugs in the treatment groups on the NCI-N87- Enhertu-resistant cell xenograft model are shown in Table 8. The results show that the ADC molecules of the present application can significantly reduce the tumor volume and have better tumor inhibition effects than the reference ADC (DS8201). Table 8: In vivo tumor growth inhibition effects of ADCs on NCI-N87- Enhertu-resistant cell transplant model Group Test drug TGI (%) Group 1 / (Vehicle) / Group 2 Reference ADC (DS8201) (3mpk) 5 Group 3 Reference ADC (DS8201) (10mpk) 55 Group 4 ADC-2 (3mpk) 17 Group 5 ADC-2 (10mpk) 94 Group 6 ADC-43 (3 mpk) 21 Test Example 9: Efficacy evaluation in human gastric cancer LD1-0017-411335 PDX tumor-bearing mice 9.1 Test objective

[0331] To evaluate the efficacy of the ADCs in the present application by investigating whether tumor growth is inhibited, delayed or cured in NU / NU mice as test animals. 9.2 Experimental steps 9.2.1 Test drugs

[0332] Blank control / or Vehicle: PBS Reference ADC (DS8201): 10 mg / kg ADC-2: 10 mg / kg 9.2.2 Preparation method: All of them were diluted with PBS. 9.2.3 Experimental method

[0333] LD1-0017-411335 human gastric cancer tumor tissue was uniformly cut into tumor fragments of approximately 3 mm x 3 mm x 3 mm (about 30-60 mg) and implanted subcutaneously into the right side of NU / NU mice. The mice were then observed after inoculation, and their weight changes and tumor growth were monitored. When the average tumor volume of the mice reached 140.81 mm3, they were randomly stratified and grouped according to the tumor size into 3 groups (2 treatment groups + 1 blank control group), with 6 mice in each group. The grouping day was defined as Day 0, and a single dose was administered via tail vein injection.

[0334] Tumor volume and body weight were measured twice a week for four weeks, and the data were recorded. Data statistics were performed using Excel 2023 statistical software: the mean value was calculated as avg; the SD value was calculated as STDEV; the SEM value was calculated as STDEV / SQRT; and the P value of the difference between groups was calculated as TTEST.

[0335] Tumor volume calculation formula: V = 0.5 a x b2 where a and b represent the long diameter and short diameter of the tumor, respectively.

[0336] The anti-tumor efficacy of the compound was evaluated by TGI (%), and the tumor growth inhibition rate was calculated using the following formula: TGI (%) = [1 - (Ti - T0) / (Ci - C0)] x 100%, where Ti is the average tumor volume of a treatment group on a given day, T0 is the average tumor volume of the same treatment group at the start of drug administration; Ci is the average tumor volume of the vehicle control group on a given day (the same day as Ti), and C0 is the average tumor volume of the vehicle control group at the start of drug administration. 9.3 Experimental results and conclusions

[0337] The schematic diagram of efficacy evaluation IN the human gastric cancer LD1-0017-411335 PDX tumor-bearing mice is shown in Fig. 10. The in vivo tumor growth inhibition (TGI) effects of the test drugs in the treatment groups on the human gastric cancer LD1-0017-411335 PDX model are shown in Table 9. The results show that the ADC-2 molecules of the present application can significantly reduce the tumor volume and have better tumor inhibition effects than the reference ADC. Table 9: In vivo tumor growth inhibition effects of ADCs on human gastric cancer LD1-0017-411335 PDX model Group Test drug TGI (%) Group 1 / (Vehicle) / Group 2 Reference ADC (DS8201) (10mpk) 66 Group 3 ADC-2 (10mpk) 105 Test Example 10: Pharmacokinetic evaluation in vivo in mice 10.1 Test objective

[0338] To evaluate the in vivo pharmacokinetic properties of ADCs in the present application in C57 mice as test animals. 10.2 Experimental steps 10.2.1 Test drugs

[0339] ADC-2: 10 mg / kg 10.2.2 Preparation method: All of them were diluted with PBS. 10.2.3 Experimental method

[0340] For each test drug, 15 C57 mice were adopted. ADC (10 mg / kg) was administered via tail vein bolus injection over approximately 1 min ± 10 s, with an injection volume of 5 mL / kg. After dosing, blood samples were collected at time points of 0.083 h, 2 h, 8 h, 24 h, 48 h, 96 h, 168 h, 336 h, 504 h, and 672 h. Serum was separated by centrifugation within 30-120 min. The free small-molecule toxin in blood samples was quantified by conventional LC-MS analysis. The concentrations of Total antibody and ADC in blood samples were quantified by conventional ELISA assays.

[0341] The PK curve of ADC-2 in mice is shown in Fig. 11, and the pharmacokinetic data are shown in Table 10. No free toxin small-molecules were detected at any time point, indicating that the ADC of the present application has good in vivo stability and good in vivo pharmacokinetic properties. Table 10: In vivo pharmacokinetic data of ADC-2 in mice PK parameters ADC-2 (Total antibody) ADC-2 (ADC) Co (pg / ml) 267.8 263.7 T1 / 2  (h) 312 231 AUCo-iast (ug.h / mL) 40610 29260 Cl (mL / h / kg) 0.1975 0.2933 Test Example 11: Efficacy evaluation in JIMT-1 tumor-bearing mice 11.1 Test objective

[0342] To evaluate the efficacy of the ADCs in the present application by investigating whether tumor growth is inhibited, delayed or cured in SCID Beige mice as test animals. 11.2 Experimental steps 11.2.1 Test drugs

[0343] Blank control / or Vehicle: PBS ADC-2: 1 mg / kg ADC-58: 1 mg / kg 11.2.2 Preparation method: All of them were diluted with PBS. 11.2.3 Experimental method

[0344] JIMT-1 cells were seeded subcutaneously in the right rib of mice. After the tumors grew for 8 days, the animals were randomly divided into groups, 6 animals per group, for a total of 3 groups (2 treatment groups + 1 blank control group). The drug was administered once by tail vein injection. Tumor volume (diameter) and body weight were measured twice a week for four weeks, and the data were recorded. Data statistics were performed using Excel 2023 statistical software: the mean value was calculated as avg; the SD value was calculated as STDEV; the SEM value was calculated as STDEV / SQRT; and the P value of the difference between groups was calculated as TTEST.

[0345] Tumor volume calculation formula: V = 0.5 a x b2 where a and b represent the long diameter and short diameter of the tumor, respectively.

[0346] The anti-tumor efficacy of the compound was evaluated by TGI (%), and the tumor growth inhibition rate was calculated using the following formula: TGI (%) = [1 - (Ti - T0) / (Ci - C0)] x 100%, where Ti is the average tumor volume of a treatment group on a given day, T0 is the average tumor volume of the same treatment group at the start of drug administration; Ci is the average tumor volume of the vehicle control group on a given day (the same day as Ti), and C0 is the average tumor volume of the vehicle control group at the start of drug administration. 11.3 Experimental results and conclusions

[0347] The in vivo tumor inhibitory effects of the treatment group on the JIMT-1 xenograft model are shown in Table 11, and the schematic diagram of the efficacy evaluation in JIMT-1 tumor-bearing mice is illustrated in Fig. 12. The results demonstrate that, compared with the control, the ADC molecule of the present application can significantly reduce the tumor volume to a greater extent. Table 11: In vivo tumor growth inhibition effects of ADCs on JIMT-1 transplant model Group Test drug TGI (%) Group 1 (Vehicle) / Group 2 ADC-2 (1mpk) 111 Group 3 ADC-58 (1mpk) 95 Test Example 12: Efficacy evaluation in NCI-N87 tumor-bearing mice 12.1 Test objective

[0348] To evaluate the efficacy of the ADCs in the present application by investigating whether tumor growth is inhibited, delayed or cured in Balb / c nude mice as test animals. 12.2 Experimental steps 12.2.1 Test drugs

[0349] Blank control / or Vehicle: PBS ADC-2: 1 mg / kg ADC-58: 1 mg / kg 12.2.2 Preparation method: All of them were diluted with PBS. 12.2.3 Experimental method

[0350] NCI-N87 cells were seeded subcutaneously in the right rib of mice. After the tumors grew for 7 days, the animals were randomly divided into groups, 6 animals per group, for a total of 3 groups (2 treatment groups + 1 blank control group). The drug was administered once by tail vein injection. Tumor volume (diameter) and body weight were measured twice a week for four weeks, and the data were recorded. Data statistics were calculated using Excel 2023 statistical software: the mean value was calculated as avg; the SD value was calculated as STDEV; the SEM value was calculated as STDEV / SQRT; and the P value of the difference between groups was calculated as TTEST.

[0351] Tumor volume calculation formula: V = 0.5 a x b2 where a and b represent the long diameter and short diameter of the tumor, respectively.

[0352] The anti-tumor efficacy of the compound was evaluated by TGI (%), and the tumor growth inhibition rate was calculated using the following formula: TGI (%) = [1 - (Ti - T0) / (Ci - C0)] x 100%, where Ti is the average tumor volume of a treatment group on a given day, T0 is the average tumor volume of the same treatment group at the start of drug administration; Ci is the average tumor volume of the vehicle control group on a given day (the same day as Ti), and C0 is the average tumor volume of the vehicle control group at the start of drug administration. 12.3 Experimental results and conclusions

[0353] The schematic diagram of efficacy evaluation in NCI-N87 tumor-bearing mice is shown in Fig. 13. The in vivo tumor growth inhibition (TGI) effects of the test drugs in the treatment groups on the NCI-N87 xenograft model are shown in Table 12. The results show that, compared with the control group, the ADC molecules of the present application can significantly reduce the tumor volume to a greater extent. Table 12: In vivo tumor growth inhibition effects of ADCs on NCI-N87 transplant model Group Test drug TGI (%) Group 1 (Vehicle) / Group 2 ADC-2 (1 mpk) 68 Group 3 ADC-58 (1 mpk) 62 Test Example 13: Efficacy evaluation in 22RV1 tumor-bearing mice 13.1 Test objective

[0354] To evaluate the efficacy of the ADCs in the present application by investigating whether tumor growth is inhibited, delayed or cured in Balb / c nude mice as test animals. 13.2 Experimental steps 13.2.1 Test drugs

[0355] Blank control / or Vehicle: PBS ADC-2: 5mg / kg ADC-57: 5 mg / kg 13.2.2 Preparation method: All of them were diluted with PBS. 13.2.3 Experimental method

[0356] 22RV1 cells were seeded subcutaneously in the right rib of mice. After the tumors grew for 7 days, the animals were randomly divided into groups, 6 animals per group, for a total of 3 groups (2 treatment groups + 1 blank control group). The drug was administered once by tail vein injection. Tumor volume (diameter) and body weight were measured twice a week for three weeks, and the data were recorded. Data statistics were calculated using Excel 2023 statistical software: the mean value was calculated as avg; the SD value was calculated as STDEV; the SEM value was calculated as STDEV / SQRT; and the P value of the difference between groups was calculated as TTEST.

[0357] Tumor volume calculation formula: V = 0.5 a x b2 where a and b represent the long diameter and short diameter of the tumor, respectively.

[0358] The anti-tumor efficacy of the compound was evaluated by TGI (%), and the tumor growth inhibition rate was calculated using the following formula: TGI (%) = [1 - (Ti - T0) / (Ci - C0)] x 100%, where Ti is the average tumor volume of a treatment group on a given day, T0 is the average tumor volume of the same treatment group at the start of drug administration; Ci is the average tumor volume of the vehicle control group on a given day (the same day as Ti), and C0 is the average tumor volume of the vehicle control group at the start of drug administration. 13.3 Experimental results and conclusions

[0359] The schematic diagram of efficacy evaluation in 22RV1 tumor-bearing mice is shown in Fig. 15. The in vivo tumor growth inhibition (TGI) effects of the test drugs in the treatment groups on the 22RV1 xenograft model are shown in Table 13. The results show that the efficacy of ADC-2 in the present application is superior to that of ADC-57. Table 13: In vivo tumor growth inhibition effects of ADCs on 22RV1 transplant model Group Test drug TGI (%) Group 1 (Vehicle) / Group 2 ADC-2 (5 mpk) 63 Group 3 ADC-57 (5 mpk) 52 Test Example 14: Stability test of compounds under antibody conjugation conditions 14.1 Test objective

[0360] To evaluate the stability of compounds under conjugation conditions and conjugation temperature by incubating the compounds in conjugation buffer. 14.2 Experimental steps

[0361] The compounds 002 and 057 (2.1 mg, dissolved in 0.1 mL of DMA) were added to 3.2 mL of 50 mM PBS (PH7.97), followed by the addition of 0.1 mL of 0.1 M EDTA and supplementation with 0.33 mL of DMA. The mixture was incubated to react at 22°C for 6 h and then at 4°C for 18 h. Samples were collected at 1 h, 2 h, 3 h, 6 h and 24 h respectively, and the purity changes of the compounds 002 and 057 were detected by HPLC. The detection results are shown in Table 14. The compound 002 exhibited superior stability in the buffer compared with the compound 057, indicating that the compound 002 possesses advantages in production process and quality control. Table 14: Stability test results of compounds in conjugation buffer Compound 1 h 2 h 3 h 6 h 24 h 002 99% 98% 95% 87% 86% 057 93% 90% 88% 76% 75% Test Example 15: Thermal stability test of ADCs 15.1 Test objective

[0362] To test the thermal stability of ADCs conjugated with different linker toxins and evaluate the effects of different linker-toxins on the thermal stability of ADCs. 15.2 Test compound

[0363] ADC-2: Formulation buffer 20 mM Histidine-HCl, 8% Sucrose, pH 5.5 ADC-57: Formulation buffer 20 mM Histidine-HCl, 8% Sucrose, pH 5.5 15.3 Experimental steps

[0364] The thermal stability of proteins was determined using the Protein Thermal Shift Dye Kie™ Kit (Cat. No. 4461146). The 1000* Protein Thermal Shift™ Dye was diluted 8-fold. A 20 pL detection system was prepared as follows: 2.5 pL / well of 8X dye, 5 pLwell of Protein Thermal Shift™ Buffer, and 12.5 pL / well of test sample. After mixing, the system was briefly centrifuged at 4000 rpm for 1 min. Detection was performed using a real-time fluorescent quantitative PCR instrument (Thermo ABI7500) with Real-Time PCR Software v2.4. The detection results are shown in Table 15. The results indicate that ADC-2 exhibits superior thermal stability compared with ADC-57. Table 15: Thermal stability test of ADCs Compound Tm (°C) ADC-2 77.4 ADC-57 76.0

[0365] The embodiments of the technical solutions of the present invention are described in an illustrative manner above. It is to be understood that the scope of protection of the present invention is not limited to the above-mentioned embodiments. Any modifications, equivalent substitutions, improvements, etc. made by those skilled in the art within the spirit and principles of the present invention should be included in the scope of protection of the claims of the present application.

Claims

1. A compound represented by formula (Ia) or (Ib), racemate, stereoisomer, tautomer, solvate,polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof: M-Z1-Tr1-L1-D (Ia) or M-Z1-L2(Tr2)-D (Ib)wherein M is a linker site to an antibody or an antigen-binding fragment thereof; preferably, M comprises a thiol-reactive group, an amino-reactive group, a carboxyl-reactive group, a proline residue-reactive group, a tyrosine residue-reactive group, a disulfide bond-bridging group, etc.; for an antibody incorporating an unnatural amino acid, M also comprises a bioorthogonal reactive group; preferably a thiol-reactive group, such as a methylsulfonylpyrimidine group, a methylsulfonylpyridine group, and a maleimide group, wherein the pyrimidine group, pyridine group, and maleimide group are optionally substituted with one or more substituents selected from the group consisting of halogen, CN, C1-6 alkyl, C1-6 alkoxy, C1-6 alkylene-O-C1-6 alkyl, and -O-(CH2CH2O)n1-CH3; n1 is an integer from 1 to 36;Z1 is a chemical bond, a divalent group, or a trivalent group substituted with a hydrophilic group, such as NH or -(CH2)qCONH-; q is an integer from 0 to 6;Tr1 is a divalent trigger group, preferably a peptide residue or a modified peptide residue, wherein the peptide residue or modified peptide residue comprises an optionally substituted natural or unnatural amino acid, an L- or D-amino acid; and it is selected from glycine-glycine-phenylalanine-glycine (GGFG), glutamic acid-valine-citrulline (EVC), valine-citrulline (VC), valine-alanine (VA), aspartic acid-valine-citrulline (DVC), glutamic acid-glycine-glycine-phenylalanine-glycine (EGGFG), aspartic acid-glycine-glycine-phenylalanine-glycine (DGGFG), and lysine-glycine-glycine-phenylalanine-glycine (KGGFG);L1 is a linking moiety between Tr1 and a bioactive molecule structural fragment D; preferably a0                      Hchemical bond, -NH-C1-6 alkyl-, -N(CH3)-Ci-6 alkyl-, H             , H                  , / nXX' 1         / 'N'^-N'irX z'n'^n'itXH                                           0 n         O 1        0 n         0  ■,,  ,,  ;Li is further preferably -NH-CH2- or H             ;D is selected from bioactive molecule structural fragments;L2 is a trivalent linking moiety connecting Z1 or M, Tr2, and the bioactive molecule structural fragment D;Tr2 is a monovalent trigger group, preferably a peptide residue, a p-glucuronide group, or a p-galactoside group;in formula (Ia) or formula (Ib), M or Z1 has at least one monovalent hydrophilic group Hp1 substituent; and / or Z1 has at least one divalent hydrophilic group Hp2 inserted or substituted, the monovalent hydrophilic group or divalent hydrophilic group is linked to other moieties of formula (Ia) or formula (Ib) via any chemical bond or linking group;the monovalent hydrophilic group Hp1 is selected from:(1) monovalent polyethylene glycol groups, preferably: -(CH2CH2O)n1-T1, -(CH2CH2O(CH2)m1CONH)n1-T1, or cyclic groups comprising -(CH2CH2O)- segments, the cyclic group optionally comprising 1 to 3 heteroatoms such as N;(2) monovalent polysarcosines: -(N(CH3)CH2CO)n2-T2;(3) monovalent polybetaines: -(CH2CH2N(CH3)(CH2COOH))n3-T3 or -(CH2CH2N(CH3)(CH2SO3H))n3-T3;(4) groups comprising carboxyl groups: -(CH2)m2-X-(CH2)m3-COOH;(5) the combinations -Hp2-Hp1 of (1)-(4) above with the following divalent hydrophilic groups are also regarded as monovalent hydrophilic groups in the present invention;the divalent hydrophilic group Hp2 is selected from divalent groups derived by removing the capping group from the above monovalent hydrophilic groups, or n4, n5 or n6 repeating units of -K(Hp1)-, preferably:(1) divalent polyethylene glycol groups, preferably: -(CH2CH2O)n1-, -(CH2CH2O(CH2)m1CONH)n1-, -[K-(CH2CH2O)n1-T1]n4-, -[K-(CH2CH2O(CH2)m1CONH)n1-T1]n4-, or cyclic groups comprising -(CH2CH2O)- segments, the cyclic group optionally comprising 1 to 3 heteroatoms such as N, for example:H. (CHjCHjOK-f-Q'(CH2CH20)n1"—I—' h2wherein n1’ and n1’’ are each an integer from 1 to 36, and q1and q2 are each an integer from 1 to 10;(2) divalent polysarcosines: -(N(CH3)CH2CO)n2- or -[K-(N(CH3)CH2CO)n2-T2]n5-;(3) divalent polybetaines: -(CH2CH2N(CH3)(CH2COOH))n3- or -[K-(CH2)m4N(CH3)2CH2COOH]n6-, or -(CH2CH2N(CH3)(CH2SO3H))n3- or -[K-(CH2)m4N(CH3)2CH2SO3H]n6-;(4) any combination of (1)-(3) above;whereinn1-n6 are each an integer from 1 to 36; m1-m5 are each an integer from 1 to 10;X is O, S or NH;K is any trivalent group, preferably a trivalent group derived from an amino acid;T1, T2 or T3 is selected from any end-capping group, preferably H, OH, C1-6 alkyl, C1-6 alkoxy, carboxyl, -(CH2)m5-CONH-(CH2)m4N(CH3)2CH2COOH, -(CH2)m4N(CH3)2CH2COOH, -C1-6alkylene-COOH, -N(C1-6 alkyl)2,, or selected from hydrophilicgroups containing monosaccharides, disaccharides, or oligosaccharides, or selected from hydrophilic groups containing multiple (2 or more) carboxyl groups, multiple (2 or more) sulfonic acid groups, or chelating groups;preferably, T1, T2 or T3 is selected from H, OH, methyl, carboxyl, -CH2CH2COOH, -N(CH2)2,OHOHY J OHOH OHT1, T2 or T3 is connected to the hydrophilic group by any linking group.

2. The compound represented by formula (Ia) or (Ib) according to claim 1, a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof, wherein Hp1 is:-(CH2CH2O)n1-H, -(CH2CH2O)n1-CH3,-(CH2)m2-X-(CH2)m3-COOH;preferably, the divalent hydrophilic group Hp2 is selected from:-(CH2CH2O)n1-,orOH3. The compound represented by formula (Ia) or (Ib), racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound thereof according to claim 1 or 2, wherein the formula (Ia) or formula (Ib) has a structure as shown in the following (Ia-1) to (Ia-6) or (Ib-1) to (Ib-6):M(Hp1)-Z1-Tr1-L1-D (Ia-1)M-Z1-Hp2-Z2-Tr1-L1-D (Ia-2)M-Z1(Hp1)-Z2-Tr1-L1-D (Ia-3)M(Hp1)-Z1-Hp2-Z2-Tr1-L1-D (Ia-4)M-Z1(Hp1)-Hp2-Z2-Tr1-L1-D (Ia-5)M(Hp1)-Z1(Hp1)-Hp2-Z2-Tr1-L1-D (Ia-6)M(Hp1)-Z1-L2(Tr2)-D (Ib-1)M-Z1-Hp2-Z2-L2(Tr2)-D (Ib-2)M-Z1(Hp1)-Z2-L2(Tr2)-D (Ib-3)M(Hp1)-Z1-Hp2-Z2-L2(Tr2)-D (Ib-4)M-Z1(Hp1)-Hp2-Z2-L2(Tr2)-D (Ib-5)M(Hp1)-Z1(Hp1)-Hp2-Z2-L2(Tr2)-D (Ib-6);wherein Z2 is a chemical bond or a divalent group, and Z1 is a divalent group or a trivalent group substituted with Hp1.

4. The compound represented by formula (Ia) or (Ib) according to any one of claims 1 to 3, a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof, wherein M is selected from the following structures:(Rb)rLg is absent or is a leaving group, and the leaving group is selected from halogen, sulfonyl, trifluoromethanesulfonyl, and methanesulfonyl;ring B is selected from 5-14-membered heteroaromatic rings, or 3-14-membered heterocyclic rings;each Rb is the same or different and is independently selected from the following groups: halogen, cyano, oxo (=O), C1-6 alkyl, halo-C1-6 alkyl, hydroxy-C1-6 alkyl, C1-6 alkoxy or C3-8 cycloalkyl, 3-8-membered heterocyclyl, C1-6 alkyl-O-C1-6 alkyl-, C1-6 alkyl-(5-6-membered) heteroaryl-, or a monovalent hydrophilic group; r is an integer from 0 to 4;Lm1 is absent, or is selected from the following groups that are unsubstituted or optionally substituted with one, two or more Rm1: C6-14 aryl, 5-14-membered heteroaryl, 3-14-membered heterocyclyl; each Rm1 is the same or different and is independently selected from H, halogen, cyano, C1-6 alkyl or HOOC-C1-3 alkylene;Lm2 is selected from the following groups that are unsubstituted or optionally substituted with one, two or more Rm2: -(CH2)s-(C=O)-, or -C=C-(CH2)t-(C=O)-; each Rm2 is the same or different and is independently selected from H, halogen, cyano, C1-6 alkyl or -C1-6 alkylene-COOH, wherein the alkylene is optionally interrupted by one, two or more of the following groups: O, NH; s and t are the same or different and are each independently an integer from 0 to 10;preferably, ring B is selected from 5-6-membered N-containing heteroaromatic rings, 3-6 membered N-containing heterocyclic rings;preferably, ring B is selected from pyrimidine rings, pyridine rings, triazine rings (such asNH .;preferably, each Rb is the same or different and is independently selected from cyano, oxo (=O),Ln'Nmethoxy, cyclopropyl, trifluoromethyl,          , / \ / o\,     '--O or       \ ;preferably, Lm1 is absent or is selected from the following groups that are unsubstituted or optionally substituted with one, two or more Rm1: phenyl, piperidinyl or piperazinyl;(  N-^     / \—preferably, Lmi is selected from \= /   ,    \— /    or    \— /    ;preferably, Lm2 is selected from the following groups that are unsubstituted or optionally substituted with one, two, or more Rm2: -CH2-(C=O)-, -(CH2)2-(C=O)-, -(CH2)5-(C=O)-, -C=C-(CH2)3-(C=O)-;preferably, M is represented by the following formula M-i:(M-i)wherein t is an integer from 0 to i0; Z is N or CR22; R2i, R22 and R23 are each independently selected from H, halogen, cyano, Ci-6 alkyl, halo-Ci-6 alkyl, hydroxy Ci-6 alkyl, Ci-6 alkoxy, C3-8 cycloalkyl, 3-8-membered heterocyclyl, Ci-6 alkyl-O-Ci-6 alkylene, or a monovalent hydrophilic group Hpi; provided that when Z is N, R2i and R23 are not both H;preferably, t is selected from i, 2, 3, 4, 5 or 6;preferably, Z is N or C-CN;preferably, R2i is selected from H, halogen, cyano, Ci-6 alkoxy, or Ci-6 alkyl-O-Ci-6 alkylene;preferably, Z is N, and R2i is selected from halogen, cyano, Ci-3 alkoxy, or Ci-3 alkyl-O-Ci-3 alkylene;preferably, Z is N, and R2i is selected from methoxy or CH3-O-CH2-;preferably, Z is C-CN, and R2i is selected from H;preferably, R23 is selected from H;5. The compound represented by formula (Ia) or (Ib) according to any one of claims 1 to 4, a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof, wherein Z1-Hp2-Z2 has the following structures:-Hp2-;-NH-(CH2)m6-HP2-;-NH-(CH2)m6-HP2-(CH2)m7CO-;-CO-(CH2)m6-HP2-;-CO-(CH2)m6-HP2-(CH2)m7CO-;-CO-(CH2)m6-HP2-(CH2)m7O(CH2)m8CO-wherein m6, m7, and m8 are each independently an integer from 0 to 10;preferably, M-Z1-Hp2-Z2 has the following structures:

6. The compound represented by formula (Ia) or (Ib) according to any one of claims 1 to 5, a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof, wherein Z1(Hp1)-Z2 has the following structures:-N(-Z6-Hp1)--N(-Z6-Hp1)-(CH2)m9CO--N(-Z6-Hp1)-(CH2)m9O(CH2)m10CO-CH2CH2CH2H2C^2    NH__Z4 HP1,HpiZ5HNwherein Z3-Z7 are chemical bonds or divalent groups; preferably, Z3-Z6 are selected from C1-10 alkylene, -O-C1-10 alkylene-, -O-C1-10 alkylene-O-, -O-C1-10 alkylene-CO-, -O-C1-10 alkylene-NH-, -NH-C1-10 alkylene-, -NH-C1-10 alkylene-O-, -NH-C1-10 alkylene-CO-, -C1-10 alkylene-CO-, or -NH-C1-10 alkylene-NH-; m9 and m10 are each an integer from 1 to 10;preferably, M-Z1(Hp1)-Z2 has the following structures:

7. The compound represented by formula (Ia) or (Ib) according to any one of claims 1 to 6, a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof, wherein Z1(Hp1)-Hp2-Z2 has the following structures: -N(Hp1)-Hp2--N(Hp1)-Hp2-(CH2)m11CO--N(Hp1)-Hp2-(CH2)m11O(CH2)m12CO-; m11 and m12 are each an integer from 1 to10;preferably, M-Z1(Hp1)-Hp2-Z2- has the following structures:0N .COOH8. The compound represented by formula (Ia) or (Ib) according to any one of claims 1 to 7, a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof, wherein Tr1 is glycine-glycine-phenylalanine-glycine (GGFG), i.e.:GGFGor it is valine-alanine (VA), i.e.:

9. The compound represented by formula (Ia) or (Ib) according to any one of claims 1 to 7, aracemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof,or a prodrug compound thereof, wherein the structure of L2 is:, wherein X1 is O or NH;preferably, Tr2 is a valine-alanine (Val-Ala) or p-glucuronide group, i.e.,preferably, the structure of M-Z1-Hp2-Z2-L2(Tr2)- is:

10. The compound represented by formula (Ia) or (Ib) according to any one of claims 1 to 9, a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof, wherein the compound has the following structures:wherein -Z1(Hp1)- is preferably:-N(-Z6-Hp1)--N(-Z6-Hp1)-(CH2)m9CO--N(-Z6-Hp1)-(CH2)m9O(CH2)m10CO-OCH2CH2Z4---HpiH2CNHwherein Z3-Z7 are chemical bonds or divalent groups; preferably, Z3-Z6 are selected from C1-10 alkylene, -O-C1-10 alkylene-, -O-C1-10 alkylene-O-, -O-C1-10 alkylene-CO-, -O-C1-10 alkylene-NH-, -NH-C1-10 alkylene-, -NH-C1-10 alkylene-O-, -NH-C1-10 alkylene-CO-, -C1-10 alkylene-CO-, or -NH-C1-10 alkylene-NH-; t2, m9 and m10 are each independently an integer from 1 to 10;Hp3 is H or Hp1, and the remaining groups are as defined in claims 1 to 9.

11. The compound represented by formula (Ia) or (Ib) according to any one of claims 1 to 10, a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof, wherein the compound has the following structures:wherein t2, n7, m13 and m14 are each independently an integer from 1 to 10; the remaining groupsare as defined in claims 1 to 10; the compound represented by formula (Ia) or formula (Ib) isfurther preferably:where m13 and m14 are each independently an integer from 1 to 10; and the remaining groups areas defined in claims 1 to 10;the compound represented by formula (Ia) or formula (Ib) is further preferably:orwherein Z, R21, R23, and n1 are as defined in any one of claims 1 to 10;preferably, n1 is an integer from 2 to 12, such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12; more preferably, n1 is an integer from 4 to 10.

12. The compound represented by formula (Ia) or (Ib) according to any one of claims 1 to 11, a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof, wherein the compound has the following structures:

013. The compound represented by formula (Ia) or (Ib), racemate, stereoisomer, tautomer,solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound thereof according toany one of claims 1 to 12, wherein preferably, D is:

14. The compound represented by formula (Ia) or (Ib) according to any one of claims 1 to 13, a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof, wherein the compound represented by formula (Ia) or formula (Ib) is of structures 001-056 or 001’-003’:L\Z15. An antibody-drug conjugate represented by formula (IIa) or (IIb) obtained from the compound represented by formula (Ia) or (Ib) according to claims 1 to 14, a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof:Ab-[M’-Zi-Tri-Li-D]p (Ila) or Ab-[M’-Zi-L2(Tr2)-D]p (Ilb)wherein Ab is an antibody or an antigen-binding fragment thereof, M' is a fragment formed by conjugating M with Ab; 0 is selected from integers or decimals between 1 and 10;preferably, Ab is an antibody or an antigen-binding fragment, the antigen-binding fragment isselected from Fab, Fab’, (Fab’)2, Fd, Fv, disulfide-linked Fv, scFv, di-scFv, (scFv)2, a diabody, and a single domain antibody (sdAb); and / or the antibody is a murine antibody, a humanized antibody, a chimeric antibody, a bispecific antibody, or a multispecific antibody;preferably, Ab is an anti-HER2 antibody or an antigen-binding fragment thereof, for example, Ab is trastuzumab or an antigen-binding fragment thereof;preferably, P is selected from integers or decimals between 4 and 9 (e.g., 7, 7.71, 7.84, 7.92, 7.94, 7.97, 7.98, 7.99, 8, 8.02, 8.06, or 8.14);preferably, the antibody-drug conjugate represented by formula (IIa) has the followingstructures:orwherein Z, R21, R23, and n1 are as defined in any one of claims 1 to 11.

16. A linker represented by formula (IIIa) or (IIIb):M-Z1-Tr1-L1’ (IIIa) or M-Z1-L2’(Tr2) (IIIb)wherein L1’ and L2’ are reactive forms of L1 and L2;preferably:when L1 is a chemical bond, L1’ is a reactive form of Tr1, for example, Tr1 is a peptide fragmenthaving a carboxyl group or an active ester at the C-terminus;when L1 is -NH-CH2-, then L1’ isHO^CH3 , where the wavy line indicates the linking siteto a peptide residue;^'Nwhen Li is H0 0-¾, then L1’ is a carbonate active ester form of p-aminobenzyl alcohol,for example, residue;, where the wavy line indicates the linking site to a peptidepreferably, the linker is:orwherein Z, R21, R23, and n1 are as defined in any one of claims 1 to 11; Y2 is selected fromhalogen, hydroxyl, or C1-6 alkoxy; preferably, Y2 is selected from hydroxyl, methoxy, ethoxy,isopropoxy, tert-butoxy.

17. A linker as shown below, being used for obtaining an antibody-drug conjugate formed bylinking an antibody to a drug via the linker,wherein Z, R21, R23, n1, n2, Tr1, L1, T1, T2, m13, and m14 are as defined in any one of claims 1 to 11; position 1 is linked to Ab, and position 2 is linked to D;preferably, the linker is selected from:OEZwherein position 1 is linked to Ab and position 2 is linked to D.

18. A pharmaceutical composition, comprising a therapeutically effective amount of the antibody-drug conjugate represented by formula (IIa) or (IIb) according to claim 15;preferably, the pharmaceutical composition further comprises one or more pharmaceuticallyacceptable excipients;preferably, the pharmaceutical composition further comprises one or more additional therapeuticagents.

19. A method for treating a tumor disease, comprising administering to a patient a prophylactically or therapeutically effective amount of at least one of the antibody-drug conjugate represented by formula (IIa) or (IIb) according to claim 15, a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof.

20. A method for treating a tumor disease, comprising administering to a patient a prophylactically or therapeutically effective amount of the pharmaceutical composition according to claim 18;preferably, the tumor disease is selected from breast cancer, gastric cancer, lung cancer, colorectal cancer, large intestine cancer, ovarian cancer, liver cancer, kidney cancer, esophageal cancer, cervical cancer, bladder cancer, pancreatic cancer, prostate cancer, nasopharyngeal carcinoma, melanoma, or leukemia;preferably, the patient includes a mammal, preferably a human.

21. Use of at least one of the antibody-drug conjugate represented by formula (IIa) or (IIb) according to claim 15, a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof, or a pharmaceutical composition thereof in the treatment of a tumor disease.

22. Use of at least one of the antibody-drug conjugate represented by formula (IIa) or (IIb) according to claim 15, a racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt thereof, or a prodrug compound thereof, or the pharmaceutical composition according to claim 18 in the manufacture of a topoisomerase I inhibitor and / or in the manufacture of a drug for preventing or treating a disease or condition associated with topoisomerase I;preferably, the disease or condition is tumor, including breast cancer, gastric cancer, lung cancer, colorectal cancer, large intestine cancer, ovarian cancer, liver cancer, kidney cancer, esophageal cancer, cervical cancer, bladder cancer, pancreatic cancer, prostate cancer, nasopharyngeal carcinoma, melanoma, or leukemia.

23. An intermediate:wherein Z, R21, R23, n1, n2, Tr1, m13, and m14 are as defined in any one of claims 1 to 11; Y1, Y3, and Y4 are the same or different and are each independently selected from halogen, hydroxyl, OSu, or C1-6 alkoxy;preferably, Y1, Y3, and Y4 are the same or different and are each independently selected from hydroxyl, methoxy, ethoxy, isopropoxy, tert-butoxy, or OSu.

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