Camptothecin compound and conjugate thereof, preparation method therefor, and use thereof

Abstract

A camptothecin compound and a conjugate thereof, or a tautomer thereof, an enantiomer thereof, a diastereomer thereof, or a mixture form thereof, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutical composition thereof, a preparation method therefor, and use thereof. The compound and the conjugate can be used for treating proliferative diseases related to abnormal cell activity.

Description

Camptothecin compounds and conjugates thereof, preparation methods and uses thereof Technical Field

[0001] The present invention belongs to the field of medical technology and relates to camptothecin compounds and conjugates thereof, their preparation methods and uses in preventing and / or treating proliferative diseases associated with abnormal cell activity (including but not limited to tumor diseases). Background Art

[0002] Camptothecin (CPT) is a cytotoxic alkaloid isolated from the plant Camptotheca acuminata of the Davidiaceae family. It can form a ternary complex with cellular DNA topoisomerase I, thereby inhibiting DNA unwinding, leading to blocked DNA replication and, in turn, cell death (Cancer Res. 1989, 49, 6365). It has broad-spectrum antiproliferative activity. However, due to its low solubility, instability, acquired tumor cell resistance and significant toxicity, it is not suitable for clinical development. Camptothecin derivatives can increase their water solubility and improve their drugability by introducing water-soluble groups or preparing prodrugs. Several camptothecin derivatives with significantly improved solubility have been approved for marketing (Med. Res. Rev. 2015, 35, 753), such as topotecan, irinotecan and belotecan, for the treatment of various types of cancer.

[0003] Camptothecin derivatives are also used to conjugate with antibodies as small molecule toxins, also known as payloads, in antibody-drug conjugates (ADCs). ADCs combine the high potency of cytotoxic small molecules with the high selectivity of antibodies for specific tumor cells. Compared to traditional chemotherapy drugs, ADCs can more precisely kill tumor cells while minimizing the effects on normal cells. In recent years, ADCs using camptothecin derivatives as small molecule toxins have made significant progress. Two camptothecin-based ADCs have been approved for cancer treatment: DS-8201a, in which the camptothecin analog Dxd is conjugated to the anti-HER2 antibody trastuzumab via a cleavable tetrapeptide-based linker, and Immu-132, in which the camptothecin analog SN-38 is conjugated to the anti-Trop-2 antibody satuzumab via a hydrolyzable, pH-sensitive linker.

[0004] However, ADCs using camptothecins or their derivatives as toxins generally have a large drug-to-antibody ratio (DAR), are difficult to manufacture, and can easily lead to ADC instability. Furthermore, camptothecin compounds often exhibit hematotoxicity, such as neutropenia, leukopenia, thrombocytopenia, and anemia, due to myelosuppression, as well as gastrointestinal side effects such as nausea, vomiting, and diarrhea.

[0005] Therefore, there is still a high clinical demand and application value in developing camptothecin compounds and their conjugates with novel structures that can enhance effectiveness and improve safety issues. Summary of the Invention

[0006] The present application provides a camptothecin compound or its tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, which may have one or more effects selected from the following groups: (1) having inhibitory activity on the proliferation of tumor cells in vitro; (2) having plasma stability; (3) having an in vivo tumor-suppressing effect; (4) having a bystander killing effect; (5) having an anti-transporter transport ability; (6) having an in vivo tumor targeting ability; and (7) having good in vivo safety.

[0007] In one aspect, the present application provides a ligand-drug conjugate, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein the ligand-drug conjugate comprises a ligand and a structure represented by Formula (Ia) or Formula (Ib):

[0008] in,

[0009] The wavy line indicates direct or indirect connection to the ligand through -NH- or X;

[0010] R 1a Selected from hydrogen, halogen, cyano, amino, hydroxyl, -C 1-6 Alkyl and -C 1-6 alkoxy;

[0011] Y is selected from -C 1-6 Alkylene, -C 2-6 Alkenylene, 3-10 membered cycloalkylene and 3-10 membered heterocyclylene; each of the alkylene, alkenylene, cycloalkylene and heterocyclylene is optionally substituted by one or more selected from hydrogen, halogen, hydroxyl, oxo, -C 1-6 Alkyl and -C 1-6 Substitution of alkoxy groups;

[0012] Z is absent or selected from -O-, -S-, -N(R 3a )-、-(C=O)-、-S(O)2-、-C 1-6 Alkylene, -OC 1-6 Alkylene, 3-10 membered cycloalkylene and 3-10 membered heterocyclylene; each of the alkylene, cycloalkylene and heterocyclylene is optionally substituted by one or more selected from hydrogen, halogen, hydroxyl, oxo, -C 1-6 Alkyl and -C 1-6 Substitution of alkoxy groups;

[0013] R 3a Selected from hydrogen, hydroxyl, -C 1-6 Alkyl, -C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, -(C=O)-C 1-6 Alkyl and -S(O)2-C 1-6 Alkyl; the alkyl, cycloalkyl and heterocycloalkyl groups are each optionally substituted by one or more radicals selected from hydrogen, halogen, hydroxy, oxo, -C 1-6 Alkyl and -C 1-6 Substitution of alkoxy groups;

[0014] R 2a Selected from hydrogen, hydroxy, amino, cyano, -C 1-6 Alkyl, -C 1-6 Alkoxy, -C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl; the alkyl, alkoxy, cycloalkyl and heterocycloalkyl are each optionally substituted by one or more selected from hydrogen, halogen, hydroxy, oxo, -C 1-6 Alkyl and -C 1-6 Substitution of alkoxy groups;

[0015] X is selected from: -O- and -N(R 3b-2 )-;

[0016] R 1b 、R 2b are independently selected from hydrogen, halogen, hydroxy, nitro, -C 1-6 Alkyl and -C 1-6 alkoxy;

[0017] or R 1b With R 2b and the atoms to which they are attached together form a 4-6 membered carbocyclic ring or a 4-6 membered heterocyclic ring;

[0018] t is an integer selected from 1 to 10;

[0019] R 3b-1 Each independently selected from hydrogen, halogen, hydroxy, amino, cyano, nitro, -C 1-6 Alkyl and -halogenated C 1-6 alkyl;

[0020] Or, any two R 3b-1 and the atoms to which it is attached together form an oxo group, a 3-6 membered carbocyclic ring or a 3-6 membered heterocyclic ring;

[0021] R 3b-2 Selected from hydrogen, hydroxyl, -C 1-6 Alkyl, -C 3-6Cycloalkyl and 3 to 6 membered heterocycloalkyl; said alkyl, cycloalkyl and heterocycloalkyl are each optionally substituted by one or more radicals selected from hydrogen, halogen, hydroxy, amino and -C 1-6 The alkyl group is substituted with a substituent.

[0022] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (Ia), wherein R 1a Selected from hydrogen, halogen, -C 1-6 Alkyl; preferably, R 1a is selected from hydrogen, halogen; further preferably, R 1a Selected from hydrogen.

[0023] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (Ia), wherein Y is selected from -C 1-6 Alkylene, 3-6 membered cycloalkylene; preferably, Y is selected from -C 1-6 Alkylene.

[0024] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (Ia), wherein Z is absent or selected from -O-, -N(R 3a )-、-S(O)2-、-C 1-6 Alkylene; preferably, Z is absent or selected from -O-.

[0025] In some embodiments, a ligand-drug conjugate comprising a ligand and a structure represented by formula (Ia) is provided, wherein R 3a Selected from hydrogen, -(C=O)-C 1-6 Alkyl, -S(O)2-C 1-6 Alkyl; preferably, R 3a Selected from -(C=O)-CH3 and -S(O)2-CH3.

[0026] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (Ia), wherein R 2a Selected from hydrogen, -C 1-6 Alkyl and -C 1-6 preferably, R 2a Selected from hydrogen and -C 1-6 alkyl.

[0027] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (Ib), wherein R 1b 、R 2b are each independently selected from hydrogen, halogen, -C 1-6 Alkoxy and -C 1-6 Alkyl; further preferably, R1b 、R 2b are each independently selected from hydrogen, F, Cl and methyl.

[0028] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (Ib), wherein R 1b With R 2b and the atoms to which it is attached together form a 5-membered heterocyclic ring; preferably, the 5-membered heterocyclic ring is

[0029] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (Ib), wherein R 3b-1 are each independently selected from hydrogen, halogen, -C 1-6 Alkyl; preferably, R 3b-1 are each independently selected from hydrogen and -C 1-6 Alkyl; further preferably, R 3b-1 are each independently selected from hydrogen and methyl.

[0030] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (Ib), wherein t is selected from an integer of 2-5; preferably, t is selected from 2 and 3.

[0031] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (Ib), wherein R 3b-2 Selected from hydrogen and -C 1-6 Alkyl; preferably, R 3b-2 Selected from hydrogen.

[0032] In some particularly preferred embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (Ia) or formula (Ib), wherein:

[0033] R 1a is selected from hydrogen, fluorine and chlorine;

[0034] Y is selected from -C 1-6 Alkylene, cyclopropylene, cyclobutylene;

[0035] Z is absent or selected from -O-, -N(R 3a )-、-S(O)2-、-C 1-6 alkylene;

[0036] R 3a Selected from hydrogen, -(C=O)-C 1-6 Alkyl, -S(O)2-C 1-6 Alkyl; preferably, R 3a Selected from -(C=O)-CH3 and -S(O)2-CH3;

[0037] R 2a Selected from hydrogen, -C 1-6 Alkyl and -C 1-6 alkyl halide;

[0038] R 1b 、R 2b are each independently selected from hydrogen, F, Cl and methyl;

[0039] Or, R 1b With R 2b and the atoms to which it is attached together form a 5-membered heterocyclic ring; preferably, the 5-membered heterocyclic ring is

[0040] t is selected from 2 and 3;

[0041] R 3b-1 are each independently selected from hydrogen and -C 1-6 alkyl;

[0042] X is selected from: -O- and -N(R 3b-2 )-;

[0043] R 3b-2 Selected from hydrogen and -C 1-6 alkyl;

[0044] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by Formula (Ia) or Formula (Ib), wherein the structure represented by Formula (Ia) and Formula (Ib) is any of the following structures:

[0045] In some embodiments, the present invention provides a ligand-drug conjugate, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein the ligand-drug conjugate comprises a ligand and a structure represented by Formula (IIa) or Formula (IIb):

[0046] in,

[0047] The wavy line indicates the L 1a linked to the ligand;

[0048] L 1a Selected from:

[0049] L 2 -(C(R L21 )2) n -;

[0050] Wherein, n is a natural number from 0 to 50, such as 0, 1, 2, 3, 4, 5, 6, 7 or 8;

[0051] L 2 Any-C(R L21 )2- units are each optionally replaced by the following structural units: -Cy-, -C(O)-, -NR L22 -, -O-, -S-, -SO-, -SO2-, -P(R L22 )-、-P(=O)(R L22 )-、-C(=S)-、-C(=NR L22 )-, -N=N-, -C=N-, -N=C-,

[0052] -Cy- is selected from phenylene, 5 to 8 membered heteroarylene, 3 to 10 membered heterocyclylene and 3 to 10 membered cycloalkylene, wherein said -Cy- is unsubstituted or independently substituted by one or more R cx replace;

[0053] R L21 、R L22 、R cx Each independently selected from hydrogen, deuterium, halogen, -NO2, -CN, -OR L2a 、-SR L2a 、-N(R L2a )2、-N + (R L2a )3, -C(O)R L2a 、-CO2R L2a 、-C(O)C(O)R L2a 、-C(O)CH2C(O)R L2a 、-S(O)R L2a 、-S(O)2R L2a 、-C(O)N(R L2a )2、-SO2N(R L2a )2、-OC(O)R L2a 、-N(R L2a )SO2R L2b 、-N(R L2a )COR L2b 、-(CH2) y -CO-(N(Me)CH2C(O)) m -OR L2a 、-(CH2) y -CO-(N(Me)CH2C(O)) m -NHR L2a 、-(CH2) y -CO-(N(Me)CH2C(O))m -N + (R L2a )3, -(CH2) y -NHCOCH2(OCH2CH2)OR L2a 、-(CH2) y -NH(COCH2(N(Me)) m -R L2a 、-(CH2) y -CONH-(CH2CH2O) m -R L2a 、-(CH2) y -NHCO-(CH2CH2O) m -R L2a 、-(CH2CH2O) m -R L2a 、-(COCH2N(Me)) m -R L2a 、-COCH2(OCH2CH2) m -OR L2a 、-CO-(CH2CH2O) m -R L2a 、-CO-(CH2) y -CONH-(CH2CH2O) m -R L2a 、-CO-(CH2) y -NHCO-(CH2CH2O) m -R L2a and R L2a Optionally substituted -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, 3-8 membered cycloalkyl, 4-10 membered heterocycloalkyl, 6-10 membered aryl, and 3-10 membered heteroaryl;

[0054] m and y are natural numbers from 0 to 50, such as 0, 1, 2, 3, 4, 5, 6, 7, or 8;

[0055] R L2a 、R L2b Each independently selected from hydrogen, deuterium, halogen, -NO2, -CN, -OH, -SH, -NH2, -N(Me)2, -CO2H, -S(O)2Me, -S(O)2OH, -C(O)NH2, -SO2NH2, -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, 3-8 membered cycloalkyl, 4-10 membered heterocycloalkyl, 6-10 membered aryl or 3-10 membered heteroaryl;

[0056] L 3 Does not exist or is an amino acid residue, a short peptide consisting of 2-10 amino acid residues, or any combination of the above groups, wherein the amino acid residue is a natural amino acid residue or a non-natural amino acid residue;

[0057] Tr does not exist or is or any combination of the above groups;

[0058] R Tr 、R Tr1 and R Tr2 Each independently selected from hydrogen, deuterium, halogen, -NO2, -CN, -OH, -SH, -NH2, -CO2H, -S(O)2OH, -C(O)NH2, -SO2NH2, -OC(O)NH2, -CH2CO-(N(Me)CH2C(O)) z -OR Tra 、-CH2CO-(N(Me)CH2C(O)) z -NHR Tra 、-(CH2CH2O) z -R Tra 、-CONH-(CH2CH2O) z -R Tra Or by R Tra Optionally substituted -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, 3-8 membered cycloalkyl, 4-10 membered heterocycloalkyl, 6-10 membered aryl or 3-10 membered heteroaryl;

[0059] R Tra For hydrogen, deuterium, halogen, -NO2, -CN, -OH, -SH, -NH2, -N(Me)2, -S(O)2Me, -CO2H, -S(O)2OH, -C(O)NH2, -SO2NH2, -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 alkynyl, 3-8 membered cycloalkyl, 4-10 membered heterocycloalkyl, 6-10 membered aryl or 3-10 membered heteroaryl,

[0060] z is independently a natural number from 0 to 50; for example, 0, 1, 2, 3, 4, 5, 6, 7 or 8;

[0061] X, Y, Z, R 1a 、R 2a 、R 1b 、R 2b 、R 3b-1and t are as described in any one of formula (Ia) or formula (Ib) of the present invention.

[0062] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (IIa) or formula (IIb), wherein L 1a Selected from: Preferably, L 1a Selected from

[0063] In some embodiments, a ligand-drug conjugate comprising a ligand and a structure represented by formula (IIa) or formula (IIb) is provided, wherein L 2 -(CHR L21 ) n -;

[0064] wherein n is selected from a natural integer from 0 to 50; for example, 0, 1, 2, 3, 4, 5, 6, 7 or 8;

[0065] L 2 Any CHR in L21 The units are each optionally replaced by the following structural units: -Cy-, -C(O)-, -NR L22 -、-O-、

[0066] -Cy- is selected from phenylene, 5- to 6-membered heteroarylene, 4- to 10-membered heterocyclylene, and 3- to 6-membered cycloalkylene, wherein said -Cy- is each independently substituted by 1 to 3 R cx replace;

[0067] R L21 、R L22 、R cx are each independently selected from hydrogen, halogen, -OR L2a 、-N(R L2a )2、-C(O)R L2a 、-S(O)2R L2a 、-C(O)N(R L2a )2、-SO2N(R L2a )2、-N(R L2a )SO2R L2b 、-N(R L2a )COR L2b 、-(CH2) y -CO-(N(Me)CH2C(O)) m -OR L2a 、-(CH2) y -CO-(N(Me)CH2C(O)) m -NHR L2a 、-(CH2)y -CONH-(CH2CH2O) m -R L2a 、-(CH2) y -NHCO-(CH2CH2O) m -R L2a 、-(CH2) y -NHCOCH2(OCH2CH2)OR L2a 、-(CH2) y -NH(COCH2(N(Me)) m -R L2a 、-(CH2) y -NHCO-(CH2CH2O) m -R L2a 、-(CH2CH2O) m -R L2a 、-(COCH2N(Me)) m -R L2a 、-COCH2(OCH2CH2) m -OR L2a 、-CO-(CH2CH2O) m -R L2a and R L2a Optionally substituted -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, 3-8 membered cycloalkyl, 4-10 membered heterocycloalkyl, 6-10 membered aryl or 3-10 membered heteroaryl;

[0068] m is a natural integer from 0 to 8;

[0069] y is selected from 0, 1, 2, 3 and 4;

[0070] R L2a 、R L2b Each is independently selected from hydrogen, halogen, -CN, -OH, -NH2, -N(Me)2, -CO2H, -C(O)NH2 and -C 1-6 alkyl.

[0071] In some embodiments, L 2 -(CH2) n -;

[0072] wherein n is selected from a natural integer from 0 to 50; for example, 0, 1, 2, 3, 4, 5, 6, 7 or 8;

[0073] L 2 Any methylene unit in each of the following structural units is optionally replaced by a 4- to 6-membered heterocyclyl group, a 3- to 6-membered cycloalkylene group, -C(O)-, -NRL22 -、-O-、

[0074] R L22 are each independently selected from hydrogen, -OR L2a 、-C(O)R L2a 、-S(O)2R L2a 、-C(O)N(R L2a )2、-SO2N(R L2a )2、-(CH2) y -CO-(N(Me)CH2C(O)) m -OR L2a 、-(CH2) y -CO-(N(Me)CH2C(O)) m -NHR L2a 、-(CH2) y -CONH-(CH2CH2O) m -R L2a 、-(CH2) y -NHCOCH2(OCH2CH2)OR L2a 、-(CH2CH2O) m -R L2a 、-(COCH2N(Me)) m -R L2a 、-COCH2(OCH2CH2) m -OR L2a 、-CO-(CH2CH2O) m -R L2a and R L2a Optionally substituted -C 1-6 alkyl;

[0075] m is a natural integer from 0 to 8;

[0076] y is 0, 1, 2, 3, or 4;

[0077] R L2a Each is independently selected from hydrogen, halogen, -CN, -OH, -NH2, -N(Me)2, -CO2H, -C(O)NH2 and -C 1-6 alkyl.

[0078] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (IIa) or formula (IIb), wherein L 2 Selected from:

[0079] in,

[0080] n1, n2, n3, n4, m are each independently selected from a natural number from 0 to 8; for example, 0, 1, 2, 3, 4, 5, 6, 7 or 8;

[0081] n5 and n6 are each independently selected from 0 or 1;

[0082] -Cy- is a 4- to 6-membered heterocyclylene or a 3- to 6-membered cycloalkylene; preferably, -Cy- is More preferably, -Cy- is

[0083] More preferably, for

[0084] More preferably,

[0085] for Among them, c and L 1a Connected, d and L 3 connected.

[0086] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (IIa) or formula (IIb), wherein Selected from:

[0087] n1, n2, n3, n4, and m are each independently selected from natural numbers from 0 to 8;

[0088] n5 and n6 are each independently selected from 0 or 1;

[0089] -Cy- is a 4- to 6-membered heterocyclylene group or a 3- to 6-membered cycloalkylene group; preferably, -Cy- is selected from: More preferably, -Cy- is selected from

[0090] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (IIa)) or formula (IIb), wherein: Selected from:

[0091] More preferably, Selected from:

[0092] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (IIa)) or formula (IIb), wherein L 3 Selected from Val, D-Val, Phe, Lys, Leu, Ile, Gly, Ala, D-Ala, Cit, Asp, Asn, Glu, Gln, Val-Cit, Val-Ala, Val- Lys, Val-Lys(Ac), Phe-Lys, Phe-Lys(Ac), Leu-Lys, Leu-Lys(Ac), Ala-Ala, Ala-Lys, D-Ala-Ala, Gly-Glu, Glu-Gly, Gly-Asp, Asp-Gly, Gly-Asn, Val-Glu, Val-Asp, Asn-Asn, Asn-Asn-Gly, Asp-Gl u, Gly-Gly-Glu, Glu-Gly-Gly, Gly-Glu-Gly, Gly-Gly-Asp, Gly-Gly-Asn, Gly-Ala-Ala, Gly-Val-A la, Gly-Val-Cit, Glu-Val-Cit, Ala-Ala-Ala, Ala-(D-Ala)-Ala, Ala-Ala-Asn, Ala-(D-Ala)-Asn ,Ala-Ala-Asp,Val-Lys-Gly,D-Val-Leu-Lys,Gly-Gly-Arg,Gly-Gly-Gly,Lys-Ala-Asn,Lys-Ala- Ala, Gly-Phe-Gly, Gly-Gly-Phe, Asn-Pro-Val, Ala-Lys-Gly, Gly-Lys-Gly, (Gly)3-Gly, (Gly)2- Phe-Gly, (Gly)2-Glu-Gly, Lys-(Ala)2-Asn, Lys-(Ala)2-Asp, (Ala)2-Pro-Val, (Ala)2-Pro-Nva, or any combination of the above fragments.

[0093] In some embodiments, L 3 Selected from Lys, Gly, Asp, Asn, Glu, Gln, Val-Cit, Val-Ala, Ala-Ala, Gly-Glu, Glu-Gly, Gly-Asp, Gly-Asn, Asn-Asn, Asn -Asn-Gly, Asp-Glu, Asp-Gly, Gly-Glu-Gly, Glu-Gly-Gly, Lys-Ala-Ala, Gly-Gly-Phe-Gly, Gly-Gly-Glu-Gly, Or any combination of the above fragments; preferably, L 3 Selected from Lys, Gly, Val-Ala, Ala-Ala, Gly-Glu, Glu-Gly, Gly-Asp, Gly-Asn, Asn-Asn, Asp-Glu, Gly-Gly-Phe-Gly, Or any combination of the above fragments; More preferably, L 3 Selected from Val-Cit, Ala-Ala, Gly-Glu, Glu-Gly, Asn-Asn, Asn-Asn-Gly, Asp-Glu, Gly-Glu-Gly, Glu-Gly-Gly, Lys-Ala-Ala, Gly-Gly-Glu-Gly, Gly-Gly-Phe-Gly and More preferably, L 3 Selected from Ala-Ala, Glu-Gly.

[0094] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (IIa) or formula (IIb), wherein: Selected from:

[0095] More preferably, Selected from

[0096] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (IIa) or formula (IIb), wherein Tr is absent or is in

[0097] R Tr 、R Tr1 and R Tr2 independently selected from hydrogen, halogen, -NO2, -CN, -OH, -NH2, -CO2H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)NH2, -CH2CO-(N(Me)CH2C(O)) z -NHMe, -(CH2CH2O) z -H, -CONH-(CH2CH2O) z -H;

[0098] z is a natural number from 0 to 8; for example, 0, 1, 2, 3, 4, 5, 6, 7, or 8.

[0099] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by formula (IIa) or formula (IIb), wherein Tr is absent or is

[0100] In some embodiments, the present invention provides a ligand-drug conjugate comprising a ligand and a structure represented by Formula (IIa) or Formula (IIb), wherein the structure represented by Formula (IIa) or Formula (IIb) is selected from:

[0101] In some embodiments, the present invention provides a ligand-drug conjugate represented by formula (IIIa) or formula (IIIb), or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof:

[0102] in:

[0103] Ab is the ligand;

[0104] q is the drug loading capacity, which is an integer or decimal from 1 to 16;

[0105] R 1a , X, Y, Z, R 2a 、R 1b 、R 2b 、R 3b-1 and t is as described in any one of the present invention (Ia) or formula (Ib);

[0106] And L 1a , L 2 , L 3 and Tr are as described in any one of the present invention (IIa) or formula (IIb).

[0107] In some embodiments, the present invention provides ligand-drug conjugates of structures represented by Formula (IIIa) and Formula (IIIb), wherein Ab is a target-binding polypeptide, antibody, or antigen-binding fragment thereof.

[0108] In some preferred embodiments, the present invention provides ligand-drug conjugates having structures represented by formula (IIIa) and formula (IIIb), wherein Ab is an antibody or an antigen-binding fragment thereof.

[0109] In some embodiments, the present invention provides ligand-drug conjugates of structures represented by formula (IIIa) and formula (IIIb), wherein the antibody Ab is selected from one or more of the following:

[0110] (1) Fully human antibodies, humanized antibodies, mouse antibodies and chimeric antibodies;

[0111] (2) Probody;

[0112] (3) Bispecific antibodies and multispecific antibodies;

[0113] (4) Monoclonal antibodies and polyclonal antibodies;

[0114] (5)IgG antibodies.

[0115] In some embodiments, the present invention provides ligand-drug conjugates of structures represented by Formula (IIIa) and Formula (IIIb), wherein in Ab, the antigen-binding fragment is selected from the group consisting of: Fab, Fab', F(ab')2, Fv, scFv, Fd, dAb, VHH, and complementarity determining region (CDR) fragments.

[0116] In some embodiments, the present invention provides ligand-drug conjugates having structures represented by formula (IIIa) and formula (IIIb), wherein Ab is a monoclonal antibody.

[0117] In some preferred embodiments, the present invention provides ligand-drug conjugates of structures represented by Formula (IIIa) and Formula (IIIb), wherein the Ab targets an antigen selected from the group consisting of HER2, HER3, B7H3, B7H4, DLL3, TROP2, Claudin18.2, GPC-3, ADAM9, CD30, CD33, CD70, and EGFR.

[0118] In some embodiments, the present invention provides ligand-drug conjugates of structures represented by Formula (IIIa) and Formula (IIIb), wherein Ab is an antibody or antigen-binding fragment thereof targeting HER2, HER3, B7H3, TROP2, Claudin18.2, GPC-3, ADAM9, CD30, CD33, CD70, GPC3, CEACAM5 and EGFR.

[0119] In some preferred embodiments, the present invention provides ligand-drug conjugates of structures represented by Formula (IIIa) and Formula (IIIb), wherein Ab is an antibody or an antigen-binding fragment thereof targeting HER3; preferably an anti-HER3 antibody or an antigen-binding fragment thereof, for example, Patritumab or a variant thereof.

[0120] In some embodiments, the ligand-drug conjugates of the structures shown in Formula (IIIa) and Formula (IIIb), wherein the anti-Her3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 with amino acid sequences as shown in SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 with amino acid sequences as shown in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, respectively.

[0121] In some embodiments, the ligand-drug conjugates of formula (IIIa) and formula (IIIb), wherein the anti-HER3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region amino acid sequence is as shown in SEQ ID NO: 7 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity thereto, and the light chain variable region amino acid sequence is as shown in SEQ ID NO: 8 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity thereto;

[0122] In some embodiments, the ligand-drug conjugates of the structures represented by Formula (IIIa) and Formula (IIIb), wherein the anti-HER3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 7, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 8.

[0123] In some embodiments, the ligand-drug conjugates of the structures shown in Formula (IIIa) and Formula (IIIb), wherein the anti-HER3 antibody or antigen-binding fragment thereof comprises the heavy chain and light chain of the antibody, wherein the amino acid sequence of the heavy chain is as shown in SEQ ID NO: 9 or a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereto, and the amino acid sequence of the light chain is as shown in SEQ ID NO: 10 or a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereto.

[0124] In some embodiments, the ligand-drug conjugates of the structures represented by Formula (IIIa) and Formula (IIIb), wherein the anti-HER3 antibody or antigen-binding fragment thereof comprises a heavy chain and a light chain of the antibody, wherein the amino acid sequence of the heavy chain is as shown in SEQ ID NO: 9, and the amino acid sequence of the light chain is as shown in SEQ ID NO: 10.

[0125] In some embodiments, the ligand-drug conjugates of the structures represented by formula (IIIa) and formula (IIIb), wherein Ab is preferably an anti-GPC-3 antibody or an antigen-binding fragment thereof, for example, codrituzumab or a variant thereof.

[0126] In some embodiments, the ligand-drug conjugates of formula (IIIa) and formula (IIIb) contain an anti-GPC-3 antibody or antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3 with amino acid sequences as shown in SEQ ID NO: 11, SEQ ID NO: 12, and SEQ ID NO: 13, respectively, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3 with amino acid sequences as shown in SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16, respectively.

[0127] In some embodiments, the ligand-drug conjugates of formula (IIIa) and formula (IIIb) comprise a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region amino acid sequence is as shown in SEQ ID NO: 17 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity thereto, and the light chain variable region amino acid sequence is as shown in SEQ ID NO: 18 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity thereto;

[0128] In some embodiments, the ligand-drug conjugates of the structures represented by Formula (IIIa) and Formula (IIIb), wherein the anti-GPC-3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 17, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 18.

[0129] In some embodiments, the ligand-drug conjugates of formula (IIIa) and formula (IIIb) are characterized in that the anti-GPC-3 antibody or antigen-binding fragment thereof comprises a heavy chain and a light chain of the antibody, wherein the amino acid sequence of the heavy chain is as shown in SEQ ID NO: 19, or a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereto, and the amino acid sequence of the light chain is as shown in SEQ ID NO: 20, or a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereto.

[0130] In some embodiments, the ligand-drug conjugates of formula (IIIa) and formula (IIIb), wherein the anti-GPC-3 antibody or antigen-binding fragment thereof comprises a heavy chain and a light chain of the antibody, wherein the amino acid sequence of the heavy chain is as shown in SEQ ID NO: 19, and the amino acid sequence of the light chain is as shown in SEQ ID NO: 20.

[0131] In some embodiments, the ligand-drug conjugates of the structures represented by Formula (IIIa) and Formula (IIIb), wherein Ab is preferably an anti-CEACAM5 antibody or an antigen-binding fragment thereof, for example, Tusamitamab or a variant thereof.

[0132] In some embodiments, the ligand-drug conjugates of formula (IIIa) and formula (IIIb) contain an anti-CEACAM5 antibody or antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3 with amino acid sequences as shown in SEQ ID NO: 21, SEQ ID NO: 22, and SEQ ID NO: 23, respectively, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3 with amino acid sequences as shown in SEQ ID NO: 24, SEQ ID NO: 25, and SEQ ID NO: 26, respectively.

[0133] In some embodiments, the ligand-drug conjugates of Formula (IIIa) and Formula (IIIb), wherein the anti-CEACAM5 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region amino acid sequence is as shown in SEQ ID NO: 27 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity thereto, and the light chain variable region amino acid sequence is as shown in SEQ ID NO: 28 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity thereto.

[0134] In some embodiments, the ligand-drug conjugates of formula (IIIa) and formula (IIIb), wherein the anti-CEACAM5 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, the amino acid sequence of the heavy chain variable region is as shown in SEQ ID NO: 27, and the amino acid sequence of the light chain variable region is as shown in SEQ ID NO: 28;

[0135] In some embodiments, the ligand-drug conjugates of Formula (IIIa) and Formula (IIIb) contain an anti-CEACAM5 antibody or antigen-binding fragment thereof, wherein the heavy chain has an amino acid sequence as shown in SEQ ID NO: 29 or a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereto, and the light chain has an amino acid sequence as shown in SEQ ID NO: 30 or a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity thereto.

[0136] In some embodiments, the ligand-drug conjugates of the structures represented by Formula (IIIa) and Formula (IIIb), wherein the anti-CEACAM5 antibody or antigen-binding fragment thereof comprises a heavy chain and a light chain of the antibody, wherein the amino acid sequence of the heavy chain is shown in SEQ ID NO: 29, and the amino acid sequence of the light chain is shown in SEQ ID NO: 30.

[0137] In some embodiments, the ligand-drug conjugates of the structures represented by Formula (IIIa) and Formula (IIIb), wherein q is an integer or decimal of 1-32, preferably an integer or decimal of 1-16, more preferably an integer or decimal of 2-8, for example, 2, 3, 4, 5, 6, 7, 8, 7.81 or 7.62.

[0138] In some embodiments, the ligand-drug conjugates of formula (IIIa) and formula (IIIb) are selected from:

[0139] wherein Ab and q are as described in any one of formula (IIIa) and formula (IIIb) of the present invention.

[0140] In some preferred embodiments, the ligand-drug conjugates represented by formula (IIIa) and formula (IIIb) are selected from:

[0141] where q is as defined above.

[0142] In another aspect, the present invention provides a compound represented by formula (IVa) or formula (IVb), or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof:

[0143] in:

[0144] R 1a , X, Y, Z, R 2a 、R 1b 、R 2b 、R 3b-1 and t is as described in any one of the present invention (Ia) or formula (Ib);

[0145] L 2 、L 3 Tr is as described in any one of formula (IIa) or formula (IIb) of the present invention;

[0146] L 1 For the connection unit.

[0147] In some embodiments, the present invention provides compounds of formula (IVa) and formula (IVb), wherein L 1 Selected from:

[0148] In some embodiments, the present invention provides compounds of formula (IVa) and formula (IVb), wherein L 1 Selected from: Preferably, L 1 Selected from

[0149] In some embodiments, the present invention provides compounds of the structures shown in Formula (IVa) and Formula (IVb), wherein, Selected from:

[0150] n1, n2, n3, n4, and m are each independently selected from natural numbers from 0 to 8;

[0151] n5 and n6 are each independently selected from 0 or 1;

[0152] -Cy- is a 4- to 6-membered heterocyclylene or a 3- to 6-membered cycloalkylene; preferably, -Cy- is selected from: More preferably, -Cy- is selected from

[0153] Preferably, Selected from: More preferably, Selected from:

[0154] In some embodiments, the present invention provides compounds of the structures shown in Formula (IVa) and Formula (IVb), wherein, Selected from:

[0155] Preferably, Selected from:

[0156] In some embodiments, the compound represented by formula (IVa) is selected from:

[0157] In some embodiments, the compound represented by formula (IVb) is selected from:

[0158] On the other hand, the present invention also provides a compound represented by formula (Va) or formula (Vb), or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof:

[0159] in,

[0160] R 3b Selected from -(C(R 3b-1 )2) t -H、-(C(R 3b-1 )2) t -OR 3b-2 、-(C(R 3b-1 )2) t -N(R 3b-2 )2、-(C(R 3b-1 )2) t -SR 3b-2 and -(C(R 3b-1 )2) t -S(O)2R 3b-2 ;

[0161] Y, Z, R 1a 、R 2a 、R 1b 、R 2b 、R 3b-1 、R 3b-2 and t are as described in any one of formula (Ia) or formula (Ib) of the present invention.

[0162] In some embodiments, the present invention provides a compound as shown in formula (Va) or formula (Vb), wherein R 3b Selected from -(C(R 3b-1 )2) t -OR 3b-2 and -(C(R 3b-1 )2) t -N(R 3b-2 )2; preferably, R 3b Selected from -(C(R 3b-1 )2) t -OR 3b-2 .

[0163] In some embodiments, the present invention provides a compound as shown in formula (Va) or formula (Vb), wherein,

[0164] R 1a selected from hydrogen;

[0165] Y is selected from -C 1-6 alkylene;

[0166] Z is absent or selected from -O-;

[0167] R 2a Selected from hydrogen and -C 1-6 alkyl;

[0168] R 1b 、R 2b are each independently selected from hydrogen, F, Cl and methyl;

[0169] Or, R 1b With R 2b and the atoms to which it is attached together form a 5-membered heterocyclic ring, wherein the 5-membered heterocyclic ring is

[0170] R 3b Selected from -(C(R 3b-1 )2) t -OR 3b-2 ;

[0171] t is selected from 2 and 3;

[0172] R 3b-1 are each independently selected from hydrogen and methyl;

[0173] R 3b-2 Selected from hydrogen.

[0174] In some embodiments, the compounds represented by formula (Va) and formula (Vb) are selected from:

[0175] Preferably, the compounds represented by formula (Va) and formula (Vb) are selected from:

[0176] The present invention also provides a pharmaceutical composition comprising substance S1 or substance S2, and one or more pharmaceutical excipients;

[0177] The substance S1 is any ligand-drug conjugate as described in the above embodiments, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or solvate;

[0178] The substance S2 is any compound as described in the above embodiments, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or solvate.

[0179] The present invention also provides a use of substance S1 or substance S2 in preparing a medicament for preventing or treating cancer;

[0180] The substance S1 is any ligand-drug conjugate as described in the above embodiments, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or solvate;

[0181] The substance S2 is any compound as described in the above embodiments, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or solvate.

[0182] Preferably, the cancer is a solid tumor or a non-solid tumor, such as esophageal cancer (e.g., esophageal adenocarcinoma and esophageal squamous cell carcinoma), brain tumor, lung cancer (e.g., small cell lung cancer and non-small cell lung cancer), squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, kidney cancer, non-Hodgkin's lymphoma, central nervous system tumor (e.g., glioma, glioblastoma multiforme, glioma or sarcoma), prostate cancer or thyroid cancer.

[0183] The present invention also provides a use of substance S1 or substance S2 in the preparation of a medicament for preventing or treating diseases associated with abnormal cell activity;

[0184] The substance S1 is any ligand-drug conjugate as described in the above embodiments, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or solvate;

[0185] The substance S2 is any compound as described in the above embodiments, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof.

[0186] Preferably, the disease associated with abnormal cell activity may be cancer; more preferably, the definition of cancer is as described above.

[0187] The present invention also provides a method for preventing or treating cancer, comprising administering an effective amount of substance S1 or substance S2 to an individual in need thereof;

[0188] The substance S1 is any of the ligand-drug conjugates described above, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof;

[0189] The substance S2 is any compound as described above, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof;

[0190] The cancer is defined as described above.

[0191] The present invention also provides a method for preventing or treating a disease associated with abnormal cell activity, the method comprising administering an effective amount of substance S1 or substance S2 to an individual in need thereof;

[0192] The substance S1 is any ligand-drug conjugate as described in the above embodiments, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or solvate;

[0193] The substance S2 is any compound as described in the above embodiments, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or solvate.

[0194] Preferably, the disease associated with abnormal cell activity may be cancer, as defined above.

[0195] In another aspect, the present invention provides a substance S for treating cancer; the substance S is any ligand-drug conjugate as described in the above embodiment or any compound as described in the above embodiment, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, or the above-mentioned pharmaceutical composition.

[0196] The disease associated with abnormal cell activity may be cancer. The definition of cancer is as described above.

[0197] Definition of terms:

[0198] Throughout this application, unless otherwise indicated, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Furthermore, the laboratory procedures for cell culture, molecular genetics, nucleic acid chemistry, and immunology used herein are conventional procedures widely used in the relevant fields. To facilitate a better understanding of this disclosure, definitions and explanations of relevant terms are provided below.

[0199] In this application, the term "pharmaceutical excipients" refers to excipients and additives used in the production of pharmaceuticals and the preparation of prescriptions. These substances, in addition to the active ingredients, have been reasonably evaluated for safety and are included in pharmaceutical preparations. In addition to providing shape, acting as carriers, and improving stability, pharmaceutical excipients also have important functions such as solubilization, dissolution assistance, and sustained-release control. They are important ingredients that may affect the quality, safety, and efficacy of drugs. Based on their source, they can be divided into natural, semi-synthetic, and fully synthetic. According to their functions and uses, they can be divided into: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, antioxidants, chelating agents, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoaming agents, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculants, filter aids, release retardants, etc. According to their route of administration, they can be divided into oral, parenteral, mucosal, transdermal or topical administration, nasal or oral inhalation administration, and ocular administration, etc. The same pharmaceutical excipient can be used in pharmaceutical preparations for different routes of administration and have different functions and uses.

[0200] In this application, the term "pharmaceutical composition" refers to various dosage forms that can be prepared according to the route of administration, such as tablets, capsules, granules, oral solutions, oral suspensions, oral emulsions, powders, tinctures, syrups, injections, suppositories, ointments, creams, pastes, ophthalmic preparations, pills, implants, aerosols, powder sprays, sprays, etc.

[0201] The pharmaceutical composition can be administered in the form of an injection, including an injection solution, sterile powder for injection, and concentrated solution for injection. Among them, the carriers and solvents that can be used include water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils such as monoglycerides or diglycerides can also be used as solvents or suspending media.

[0202] In this application, the term "pharmaceutically acceptable salt" or "pharmaceutically usable salt" generally refers to a salt of a compound or ligand-drug conjugate of the present application, which salt may be safe and / or effective when used in mammals and may have the desired biological activity.

[0203] The term "drug loading" generally refers to the average amount of cytotoxic drug loaded per ligand and can also be expressed as the ratio of the amount of cytotoxic drug to the amount of antibody, for example, the drug / antibody ratio (DAR). The cytotoxic drug loading can range from 0 to 20, as an integer or decimal. In embodiments of the present application, the drug loading is expressed as q, which can be, for example, an integer or decimal of 1-2, 2-3, 3-4, 4-5, 5-6, 6-7, 7-8, 8-9, or 9-10. For example, the drug loading q is 7.8 or 7.9. The drug loading of each ADC molecule after the coupling reaction can be characterized by conventional methods such as UV / visible spectroscopy, mass spectrometry, HIC, ELISA assay, and HPLC.

[0204] In this application, the term "ligand-drug conjugate" generally refers to a ligand linked to a biologically active cytotoxic drug via a stable linker. In some embodiments of this application, the "ligand-drug conjugate" may be an antibody-drug conjugate (ADC), which may be a monoclonal antibody or antigen-binding fragment linked to a biologically active cytotoxic drug via a stable linker.

[0205] In this application, the term "ligand" generally refers to small molecules, polypeptides, RNA, DNA, carbohydrates and macromolecular compounds that can recognize and bind to antigens or receptors associated with target cells. The function of the ligand can be to present the drug to the target cell population bound to the ligand. These ligands include but are not limited to protein hormones, lectins, growth factors, antibodies or other molecules that can bind to cells, receptors and / or antigens. In this application, the ligand can be represented by Ab, and the ligand antigen forms a connection bond with a connecting unit (also called "linker" or "linker") through a heteroatom on the ligand. The ligand can be an antibody or an antigen-binding fragment thereof. The antibody can be selected from a chimeric antibody, a humanized antibody, a fully human antibody or a murine antibody; the antibody can be a monoclonal antibody. For example, the antibody can be an antibody targeting a target selected from the following targets: HER2, HER3, B7H3, TROP2, Claudin 18.2, CD30, CD33, CD70 and EGFR. For example, the antibody can be an antibody targeting a target selected from the group consisting of 5T4, AGS-16, ANGPTL4, ApoE, CD19, CTGF, CXCR5, FGF2, MCPT8, MFI2, MS4A7, NCA, Sema5b, SLITRK6, STC2, TGF, 0772P, 5T4, ACTA2, ADGRE1, AG-7, AIF1, AKR1C1, AKR1C2, ASLG659, Axl, B7H3, BAFF-R, BCMA, BMPR1B, BNIP3, C1QA, C1QB, CA6, CADM1, CCD79b, CCL5, CCR5, CCR7, CD11c, CD123, CD138, CD142, CD147, CD166, CD19, CD19,CD22, CD21, CD20, CD205, CD22, CD223, CD228, CD25, CD30, CD33, CD37, CD38, CD40, CD45, CD45(PTPRC), CD46, CD47, CD49D(ITGA4), CD56, CD66e, CD70, CD71, CD72, CD74, CD79a, CD79b, CD80, CDCP1, CDH11, CDllb, CEA, CEACAM5, c-Met, COL6A3, COL7A1, CRIPTO, CSF1R, CTSD, CTSS, CXCL11, CXCL10, DDIT4 , DLL3, DLL4, DR5, E16, EFNA4, EGFR, EGFRvIII, EGLN, EGLN3, EMR2, ENPP3, EpCAM, EphA2, EphB2R, ETBR, FcRH2, FcRHl, FGFR2, FGFR3, FLT3, FOLR-α, GD2 , GEDA, GPC-1, GPC-3, GPNMB, GPR20, GZMB, HER2, HER3, HLA-DOB, HMOX1, IFI6, IFNG, IGF-1R, IGFBP3, IL10RA1, IL-13R, IL-2, IL20Ra, IL-3, IL-4, IL-6, IRTA2, KISS1R, KRT33A, LIV-1, LOX, LRP-1, LRRC15, LUM, LY64, LY6E, Ly86, LYPD3, MDP, MMP10, MMP14, MMP16, MPF, MSG783, MSLN, MUC-1, NaPi2b, Napi3b, Nectin-4, NOG, P2X5, pCAD, P-Cadherin, PDGFRA, PDK1, PD-L1, PFKFB3, PGF, PGK1, PIK3AP1, PIK3CD, PLOD2, PSCA, PSCAhlg, PSMA, PTK7, P-Cadherin, RN F43, ROR1, ROR2, SERPINE1, SLC39A6, SLTRK6, STAT1, STEAP1, STEAP2, TCF4, TENB2, TGFB1, TGFB2, TGFBR1, TNFRSF21, TNFSF9, Trop-2, TrpM4, Tyro7, UPK1B, VEGFA, WNT5A, ADAM9, epidermal growth factor, brevican, mesothelin, sodium phosphate cotransporter 2B, Claudin18.2, endothrin receptor, mucins (such as mucin 1 and mucin 16), guanylate cyclase C, integrin α4β7, integrin αvβ6, trophoblast glycoprotein, and tissue factor.

[0206] In this application, the term "antibody or its antigen-binding fragment" generally refers to an immunological binder, which extends to all antibodies from all species, including dimers, trimers and multimer antibodies; bispecific antibodies; chimeric antibodies; fully human antibodies; humanized antibodies; recombinant and reconstructed antibodies and their fragments. The term "antibody or its antigen-binding fragment" can refer to any antibody-like molecule with an antigen-binding region, and the term includes small molecule fragments such as Fab', Fab, F(ab')2, single domain antibodies (DABs), Fv, scFv (single chain Fv), linear antibodies, diabodies, etc. The term "antigen-binding fragment" can refer to one or more fragments of an antibody that retain the ability to specifically bind to an antigen. For example, a fragment of a full-length antibody can be used to implement the antigen-binding function of an antibody. The techniques for preparing and using various antibody-based constructs and fragments are well known in the art. The antibodies may include: anti-HER2 (ErbB2) antibody, anti-EGFR antibody, anti-B7H3 antibody, anti-c-Met antibody, anti-HER3 (ErbB3) antibody, anti-HER4 (ErbB4) antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD44 antibody, anti-CD56 antibody, anti-CD70 antibody, anti-CD73 antibody, anti-CD105 antibody, anti-CEA antibody, anti-GPC3 antibody, anti-A33 antibody, anti-Cripto antibody, anti-EphA2 antibody, anti-G250 antibody, anti-MUCl antibody, anti-Lewis Y antibody, anti-TROP2 antibody, anti-Claudin antibody 18.2 antibody, anti-VEGFR antibody, anti-GPNMB antibody, anti-Integrin antibody, anti-PSMA antibody, anti-Tenascin-C antibody, anti-SLC44A4 antibody, anti-ADAM9 antibody and anti-Mesothelin antibody, such as Patritumab and / or Codrituzumab and / or Tusamitamab.

[0207] In this application, the term "chimeric antibody" generally refers to an antibody formed by fusing the variable region of a murine antibody with the constant region of a human antibody, which can reduce the immune response induced by the murine antibody. Methods for creating chimeric antibodies include, for example, establishing a hybridoma that secretes a murine-specific monoclonal antibody, cloning the variable region genes from the murine hybridoma cells, cloning the human antibody constant region genes as needed, and then ligating the murine variable region genes with the human constant region genes to form a chimeric gene, which is then inserted into an expression vector. Chimeric antibody molecules can then be expressed in eukaryotic or prokaryotic systems.

[0208] In this application, the term "humanized antibody", also referred to as CDR-grafted antibody, generally refers to an antibody produced by transplanting mouse CDR sequences into a human antibody variable region framework, i.e., into different types of human germline antibody framework sequences. Humanized antibodies can overcome the problem of chimeric antibodies inducing strong heterologous reactions due to carrying a large amount of mouse protein components. Such framework sequences can be obtained from public DNA databases or published references including germline antibody gene sequences. For example, the germline DNA sequences of human heavy chain variable region and light chain variable region genes can be found in the "VBase" human germline sequence database.

[0209] In this application, the terms "fully human antibody", "human antibody", "fully human antibody" or "completely human antibody" are used interchangeably, and both the variable and constant regions of the antibody may be of human origin, eliminating immunogenicity and toxic side effects.

[0210] The antibodies or ligands described herein may be fully human monoclonal antibodies. Relevant technologies for preparing fully human antibodies may include: human hybridoma technology, EBV-transformed B lymphocyte technology, phage display technology, transgenic mouse antibody preparation technology, and single B cell antibody preparation technology.

[0211] In this application, the term "CDR" generally refers to one of the six hypervariable regions within the variable domain of an antibody that primarily contributes to antigen binding. The most commonly used definitions of the six CDRs are provided, for example, by Kabat EA et al. (1991) Sequences of proteins of immunological interest. NIH Publication 91-3242), Chothia et al., "Canonical Structures For the Hypervariable Regions of Immunoglobulins," J. Mol. Biol. 196: 901 (1987); and MacCallum et al., "Antibody-Antigen Interactions: Contact Analysis and Binding Site Topography," J. Mol. Biol. 262: 732 (1996). As used in this application, the Kabat definition of CDRs can be applied to CDR1, CDR2 and CDR3 of the light chain variable domain (CDRL1, CDRL2, CDRL3 or L1, L2, L3), and CDR1, CDR2 and CDR3 of the heavy chain variable domain (CDR H1, CDRH2, CDRH3 or H1, H2, H3).

[0212] The term "one or more" or the similar expression "at least one" may mean, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more;

[0213] When the lower limit and upper limit of a numerical range are disclosed, any value and any included range falling within the range is specifically disclosed. In particular, every range of values ​​disclosed herein should be understood to mean every value and range encompassed within the broader range;

[0214] For example, the statement "C 1-6 " should be understood to include any sub-ranges therein and each point value, such as C 2-5 、C 3-4 、C 1-2 、C 1-3 、C 1-4 、C 1-5 etc., as well as C1, C2, C3, C4, C5, C6, etc. For example, the expression “C 3-10 ” should also be understood in a similar manner, for example, any sub-ranges and point values ​​contained therein may be included, for example, C 3-9 、C 6-9 、C 6-8 、C6-7 、C 7-10 、C 7-9 、C 7-8 、C 8-9 etc. and C3, C4, C5, C6, C7, C8, C9, C 10 etc. For another example, the expression "3-10 yuan" should be understood to include any sub-ranges and point values ​​therein, such as 3-4 yuan, 3-5 yuan, 3-6 yuan, 3-7 yuan, 3-8 yuan, 3-9 yuan, 4-5 yuan, 4-6 yuan, 4-7 yuan, 4-8 yuan, 5-7 yuan, 5-8 yuan, 6-7 yuan, etc., as well as 3, 4, 5, 6, 7, 8, 9, 10 yuan, etc. For another example, the expression "5-10 yuan" should also be understood in a similar manner, such as it can include any sub-ranges and point values ​​contained therein, such as 5-6 yuan, 5-7 yuan, 5-8 yuan, 5-9 yuan, 5-10 yuan, 6-7 yuan, 6-8 yuan, 6-9 yuan, 6-10 yuan, 7-8 yuan, etc., as well as 5, 6, 7, 8, 9, 10 yuan, etc.

[0215] In the present application, the term "natural number" is, for example, 0-50, 0-40, 0-30, 0-20, 0-10, 0-8, 0-6, 0-6, 0-4 or 0-2; or 0, 1, 2, 3, 4, 5, 6, 7 or 8.

[0216] In this application, the term "alkyl" refers to a saturated straight or branched hydrocarbon group. 1-6 "Alkyl" refers to a saturated straight or branched chain hydrocarbon group having 1 to 6 carbon atoms (e.g., 1, 2, 3, 4, 5 or 6 carbon atoms). 1-6 The term "alkyl" refers to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl or n-hexyl.

[0217] In this application, the term "C 0-6 In the case of "alkylene", when C0 alkylene is 0, it is a connecting bond. For example, in the compounds of formula (Va) and formula (Vb) of the present invention, when L 2 -(CH2) n - and n = 0, L 2 Indicates a connection key.

[0218] In this application, the term "C 0-6 In the case of "alkyl", when C0 alkyl is 0, it is hydrogen. For example, in the compound of formula (Ib) of the present invention, when R 3b Yes-(C(R 3b-1 )2) t -H and t=0, R 3b Represents hydrogen.

[0219] In this application, the term "alkylene" refers to a saturated straight-chain or branched divalent hydrocarbon group. For example, the term "C 1-6 "Alkylene" refers to a saturated straight or branched divalent hydrocarbon group having 1 to 6 carbon atoms. 1-6 Examples of "alkylene" include, but are not limited to, methylene, ethylene, propylene, or butylene.

[0220] In this application, the term "alkenyl" refers to a straight-chain or branched aliphatic hydrocarbon group having one or more carbon-carbon double bonds. 2-6 The term "alkenyl" refers to an alkenyl group having 2 to 6 carbon atoms and one, two or three (preferably one) carbon-carbon double bonds (e.g., ethenyl, 1-propenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl, 4-methyl-3-pentenyl, etc.).

[0221] In this application, the term "alkynyl" refers to a straight-chain or branched aliphatic hydrocarbon group having one or more carbon-carbon triple bonds. 2-6 The term "alkynyl" refers to an alkynyl group having 2 to 6 carbon atoms and one, two or three (preferably one) carbon-carbon triple bonds (e.g., ethynyl, 1-propynyl, 2-propynyl, 2-butynyl, 3-butynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl, etc.).

[0222] In this application, the term "aryl" refers to a monocyclic or condensed aromatic hydrocarbon group having a conjugated π electron system. For example, the term "C 6-10 The term "aryl" refers to an aromatic group having 6 to 10 carbon atoms (e.g., phenyl, naphthyl, etc.).

[0223] As used herein, the term "heteroaryl" or "heteroaromatic ring" refers to monocyclic and fused heterocyclic ring systems having one or more conjugated π-electron systems, wherein one or more (e.g., 1, 2, or 3) ring atoms are heteroatoms selected from N, O, P, and S, and the remaining ring atoms are C. A heteroaryl group or heteroaromatic ring can be characterized by the number of ring atoms. For example, a 5-12 membered heteroaryl group can contain 5-12 (e.g., 5, 6, 7, 8, 9, 10, 11, or 12) ring atoms, particularly 5, 6, 9, or 10 ring atoms. Examples of heteroaryl groups include thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, pyridinyl, pyrazinyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, indolyl, and the like.

[0224] In this application, the term "cycloalkyl" refers to a saturated or partially saturated, monocyclic or polycyclic (such as bicyclic) non-aromatic hydrocarbon group. For example, "C 3-12 "Cycloalkyl" or "3-12 membered cycloalkyl" refers to a cycloalkyl group having 3-12 ring carbon atoms (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12). Common cycloalkyl groups include, but are not limited to, monocyclic cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclobutene, cyclopentene, cyclohexene, etc.; or bicyclic cycloalkyl groups, including fused rings, bridged rings or spiro rings, such as bicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl, bicyclo[5.2.0]nonyl, decahydronaphthyl, etc.

[0225] In this application, the term "cycloalkylene" refers to a saturated or partially saturated, monocyclic or polycyclic (such as bicyclic) non-aromatic divalent cyclic group. For example, "C 3-12 "Cycloalkylene" or "3-12 membered cycloalkylene" refers to a cycloalkylene group having 3-12 ring carbon atoms (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12). Common cycloalkylene groups include, but are not limited to, monocyclic cycloalkylene groups such as cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, cycloheptylene, cyclobutene, cyclopentene, cyclohexene, etc.; or bicyclic cycloalkylene groups including fused, bridged or spiro rings such as bicyclo[1.1.1]pentylene, bicyclo[2.2.1]heptylene, bicyclo[3.2.1]octylene, bicyclo[5.2.0]nonylene, decahydronaphthylene, etc.

[0226] The term "heterocycloalkyl" refers to a saturated or partially saturated non-aromatic cyclic group containing at least one heteroatom selected from N, O, P, and S. Preferably, the number of heteroatoms is 1, 2, 3, or 4. Examples include 3-8-membered and 3-6-membered heterocycloalkyl groups. Specific examples include, but are not limited to, oxiranyl, oxocyclobutane, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, and homopiperazinyl.

[0227] The term "heterocycloalkylene" refers to a saturated or partially saturated, non-aromatic, divalent cyclic group containing at least one ring member selected from N, O, P, and S, wherein the number of heteroatoms is preferably 1, 2, 3, or 4. Examples include 3-8 membered and 3-6 membered heterocycloalkylene groups. Specific examples include, but are not limited to, oxiranylene, oxocyclobutanylene, pyrrolidinylene, tetrahydrofuranylene, piperidinylene, piperazinylene, tetrahydropyranylene, and homopiperazinylene.

[0228] The term "fused ring (fused ring system)" refers to a chemically feasible polycyclic structure formed by two or more (e.g., 3, 4, or 5) carbocyclic or heterocyclic rings with shared ring edges or shared atoms, wherein the carbocyclic ring includes a cycloalkyl group and an aryl group, and the heterocyclic ring includes a heteroaromatic ring and a heterocycloalkyl group. The fused ring system includes, but is not limited to, a fused ring system formed by a cycloalkyl group and a cycloalkyl group, a fused ring system formed by a cycloalkyl group and a heterocycloalkyl group, a fused ring system formed by a cycloalkyl group and an aromatic ring, a fused ring system formed by a cycloalkyl group and a heteroaromatic ring, a fused ring system formed by a heterocycloalkyl group and a heteroaromatic ring, a fused ring system formed by a heterocycloalkyl group and an aromatic ring, a fused ring system formed by a heteroaromatic ring and a heteroaromatic ring, a fused ring system formed by a heteroaromatic ring and an aromatic ring, and the like.

[0229] In this application, the term "halogen" generally refers to fluorine, chlorine, bromine, iodine, for example fluorine, chlorine.

[0230] In this application, the term "each independently" means that at least two groups (or fragments) present in a structure with the same or similar value ranges can have the same or different meanings in specific circumstances. For example, if substituent X and substituent Y are each independently hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl, then when substituent X is hydrogen, substituent Y can be either hydrogen, or halogen, hydroxyl, cyano, alkyl, or aryl; similarly, when substituent Y is hydrogen, substituent X can be either hydrogen, or halogen, hydroxyl, cyano, alkyl, or aryl.

[0231] In this application, the term "optional" or "optionally" generally means that the subsequently described event or circumstance can but need not occur, and the description includes instances where the event or circumstance occurs or does not occur. For example, "a heterocyclic group optionally substituted with an alkyl group" means that the alkyl group can but need not be present, and the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

[0232] In the present application, the term "substituted" and its other variant forms in this article refer to that one or more (such as 1, 2, 3 or 4) atoms or atomic groups (such as hydrogen atoms) on the specified atom are replaced by other equivalents, provided that the normal valence of the specified atom or atomic group in the current situation is not exceeded, and a stable compound can be formed. If a certain atom or atomic group is described as "optionally substituted by ... ", it can be substituted or unsubstituted. Unless otherwise indicated, the attachment site of a substituent herein can be from any suitable position of a substituent. When the link in a substituent is shown as a chemical bond between two atoms connected to each other in a ring system, it means that the substituent can be connected to any ring-forming atom in the ring system.

[0233] This article uses wavy lines The bonds in the structural formulae represented are intended to indicate that the structure represents either a cis or trans isomer, or a mixture of cis and trans isomers in any ratio.

[0234] The term "oxo," as used herein alone or in combination with other groups, refers to =0.

[0235] In the present application, one or more hydrogen atoms in a group, for example up to 5, for example 1 to 3 hydrogen atoms, are independently replaced by a corresponding number of substituents. The substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) whether substitution is possible or not without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[0236] In this application, the term "amino acid" includes natural amino acids and non-natural amino acids, and the writing of conventional amino acids follows conventional usage. See, for example, Immunology-A Synthesis (2nd Edition, ES Golub and DRGren, Eds., Sinauer Associates, Sunderland, Mass. (1991)), which is incorporated herein by reference. In this article, the terms "polypeptide" and "protein" have the same meaning and are used interchangeably. And in this application, amino acids are generally represented by single-letter and three-letter abbreviations known in the art. For example, alanine can be represented by A or Ala; arginine can be represented by R or Arg; glycine can be represented by G or Gly; glutamine can be represented by Q or Gln;

[0237] As used herein, the term "unnatural amino acid" has the following structure: wherein r is selected from 0, 1, 2, 3, 4 and 5; wherein R a 、R b Each independently selected from -C 1-6 Alkylene-NH2, -C 1-6 Alkylene-NH-C 1-6 Alkyl, -C 1-6 Alkylene-N(C 1-6 Alkyl)2, -C 1-6 Alkylene-NH-C 3-10 Cycloalkyl, -C 1-6 Alkylene-N(3-10 membered cycloalkyl)(C 1-6 Alkyl), -C 1-6 Alkylene-C 3-10 Cycloalkyl, -C 1-6 Alkylene-(3-10 membered heterocycloalkyl), -C 1-6Alkylene-NHCOC 1-6 Alkyl, -C 1-6 Alkylene-NHCOOC 1-6 Alkyl, -C 1-6 Alkylene-NHS(O)2C 1-6 Alkyl, -C 1-6 Alkylene-S(O)2-C 1-6 Alkyl, -C 1-6 Alkylene-S(O)2-C 3-10 Cycloalkyl, -C 1-6 Alkylene-S(O)2-NH2, -C 1-6 Alkylene-COOH, -C 1-6 Alkylene-CONH2, -C 1-6 Alkylene-CONHC 1-6 Alkyl, -C 1-6 Alkylene-CO (3-10 membered heterocycloalkyl), and The alkyl, alkylene, cycloalkyl, and heterocycloalkyl groups are each independently optionally substituted by one or more substituents selected from H, halogen, -OH, -NH2, -SH, -NO2, CN, -COOH, and oxo; or any R a 、R b Together with the atoms to which it is attached, it forms a 3-10 membered heterocycloalkyl or a 3-10 membered cycloalkyl; each of the cycloalkyl and heterocycloalkyl groups is optionally substituted with one or more substituents selected from H, halogen, -OH, -NH2, -SH, -NO2, CN, -COOH and oxo;

[0238] In this application, the term "compound" generally refers to a substance having two or more different elements. For example, the compound of the present application can be an organic compound, for example, the compound of the present application can be a compound with a molecular weight of 500 Daltons or less, a compound with a molecular weight of 1000 Daltons or less, a compound with a molecular weight of 1000 Daltons or more, or a compound with a molecular weight of 1000 Daltons or more, or a compound with a molecular weight of 10000 Daltons or more, or a compound with a molecular weight of 100000 Daltons or more. In this application, a compound can also refer to a compound connected by chemical bonds, for example, a compound in which one or more molecules with a molecular weight of 1000 Daltons or less are connected to a biomacromolecule by chemical bonds, and the biomacromolecule can be a polysaccharide, protein, nucleic acid, polypeptide, etc. For example, the compound of the present application can include a compound in which a protein is connected to one or more molecules with a molecular weight of 1000 Daltons or less, a compound in which a protein is connected to one or more molecules with a molecular weight of 10000 Daltons or less, or a compound in which a protein is connected to one or more molecules with a molecular weight of 100000 Daltons or less.

[0239] In this application, the term "stereoisomer" refers to an isomer formed by at least one asymmetric center. In compounds with one or more (e.g., one, two, three, or four) asymmetric centers, racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers can be produced. Specific individual molecules can also exist as geometric isomers (cis / trans). Similarly, the compounds of the present invention can exist as mixtures of two or more structurally different forms in rapid equilibrium (commonly referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-ketone tautomers, nitroso-oxime tautomers, imine-enamine tautomers, etc. It is to be understood that the scope of this application encompasses all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%).

[0240] In this article, solid lines can be used Solid wedge or virtual wedge The carbon-carbon bonds of the compounds of the present invention are depicted. The use of solid lines to depict bonds to asymmetric carbon atoms is intended to indicate that all possible stereoisomers at that carbon atom are included (e.g., specific enantiomers, racemic mixtures, etc.). The use of solid or dashed wedges to depict bonds to asymmetric carbon atoms is intended to indicate that the indicated stereoisomers exist. When present in a racemic mixture, solid and dashed wedges are used to define relative stereochemistry, not absolute stereochemistry. Unless otherwise indicated, the compounds of the present invention are intended to exist as stereoisomers, including cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotational isomers, conformational isomers, atropisomers, and mixtures thereof. The compounds of the present invention may exhibit more than one type of isomerism and consist of mixtures thereof (e.g., racemic mixtures and diastereomeric pairs).

[0241] It will be understood by those skilled in the art that the structural formulas used in the present invention to describe groups are based on the conventions used in the art. This means that the corresponding group R is connected to other fragments or groups in the compound through this site. A "-" at the end of a group indicates that the group is connected to another fragment in the molecule through this site. For example, CH3-C(=O)- means that the C(=O) in the acetyl group is connected to another fragment in the molecule.

[0242] In this application, the term "comprising" generally refers to including the features specified but not excluding other elements. The terms "above" and "below" generally refer to including the number.

[0243] Unless otherwise indicated, the structures described herein may also include compounds that differ only in the presence or absence of one or more isotopically enriched atoms. For example, compounds identical to the structures described herein except for the replacement of a hydrogen atom by deuterium or tritium, or the replacement of a carbon atom by carbon-13 or carbon-14, are within the scope of this application.

[0244] The terms "active ingredient," "therapeutic agent," "active substance," or "active agent" refer to a chemical entity that is effective in treating one or more symptoms of a target disorder or condition.

[0245] As used herein, unless otherwise indicated, the terms "treat," ...

[0246] As used herein, "subject" includes humans and non-human animals. Exemplary human subjects include human subjects suffering from diseases (e.g., the diseases described herein) (referred to as patients) or normal individuals. "Non-human animals" herein include all vertebrates, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, livestock and / or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.).

[0247] Those skilled in the art will appreciate that, since nitrogen requires an available lone pair of electrons to be oxidized to oxides, not all nitrogen-containing heterocycles can form nitrogen oxides. Those skilled in the art will recognize nitrogen-containing heterocycles that can form nitrogen oxides. Those skilled in the art will also recognize that tertiary amines can form nitrogen oxides. Synthetic methods for preparing nitrogen oxides of heterocycles and tertiary amines are well known to those skilled in the art and include oxidizing heterocycles and tertiary amines with peroxyacids such as Peracetic Acid and Metachloroperbenzoic Acid (m-CPBA), hydrogen peroxide, alkyl hydroperoxides such as tert-butyl hydroperoxide, sodium perborate, and dioxirane such as dimethyldioxirane. These methods for preparing nitrogen oxides have been extensively described and reviewed in the literature, see for example: TL Gilchrist, Comprehensive Organic Synthesis, vol. 7, pp 748-750 (AR Katritzky and AJ Boulton, Eds., Academic Press); and GWH Cheeseman and ESGWerstiuk, Advances in Heterocyclic Chemistry, vol. 22, pp 390-392 (AR Katritzky and AJ Boulton, Eds., Academic Press).

[0248] Those skilled in the art will understand that the compounds covered by the present invention are all chemically feasible compounds; and all chemical bonds are connected in chemically feasible ways.

[0249] The present invention also encompasses compounds of the present invention that contain protecting groups. During any process for preparing the compounds of the present invention, it may be necessary and / or desirable to protect sensitive or reactive groups on any of the molecules involved, thereby forming a chemically protected form of the compounds of the present invention. This can be achieved using conventional protecting groups, for example, those described in TW Greene & P.G.M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 2006, which references are incorporated herein by reference. Protecting groups can be removed at an appropriate subsequent stage using methods known in the art.

[0250] The present invention also encompasses methods for preparing the compounds described herein. It should be understood that the compounds of the present invention can be synthesized using the methods described below, as well as synthetic methods known in the art of synthetic organic chemistry or variations thereof known to those skilled in the art. Preferred methods include, but are not limited to, those described below. The reaction can be carried out in a solvent or solvent mixture that is appropriate for the reagents and materials used and suitable for the transformation to be achieved.

[0251] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention.

[0252] The reagents and raw materials used in the present invention are commercially available.

[0253] The positive progress of the present invention is that the compounds of the present invention have one or more of the following advantages:

[0254] (1) It has inhibitory activity on the proliferation of tumor cells in vitro;

[0255] (2) Having plasma stability;

[0256] (3) Has tumor-suppressing effect in vivo;

[0257] (4) Has a bystander effect;

[0258] (5) possessing anti-transporter transport capability;

[0259] (6) Possesses the ability to target tumors in vivo;

[0260] (7) Good in vivo safety;

[0261] In addition, the coupling method disclosed herein has a wide range of applications and can be widely used for coupling with bioactive molecules such as antibodies or targeted small molecule ligands. In summary, the protein degraders, linkers, antibodies and ADCs of the present invention have significant clinical value. DETAILED DESCRIPTION

[0262] The present invention includes all combinations of the described specific embodiments. Further embodiments of the present invention and the full scope of applicability will become apparent from the detailed description provided below. However, it should be understood that although the detailed description and specific examples indicate preferred embodiments of the present invention, these descriptions and examples are provided by way of illustration only, because various changes and modifications within the spirit and scope of the present invention will become apparent to those skilled in the art from this detailed description. For all purposes, all publications, patents and patent applications cited herein, including citations, will be incorporated herein by reference in their entirety. The present invention is further illustrated below by way of example, but the present invention is not limited to the scope of the described embodiments. The experimental methods for which specific conditions are not specified in the following examples are selected according to conventional methods and conditions, or according to the product specifications.

[0263] Mass spectrometry (MS) was measured using an Agilent (ESI) mass spectrometer, manufactured by Agilent, model: Agilent 6120B.

[0264] Preparative high performance liquid chromatography (HPLC) was performed using a Shimadzu LC-8A preparative liquid chromatograph (YMC, ODS, 250 × 20 mm column).

[0265] Thin layer chromatography purification was performed using GF 254 (0.4-0.5 nm) silica gel plates produced in Yantai.

[0266] The reaction is monitored by thin layer chromatography (TLC) or liquid chromatography-mass spectrometry (LC-MS). The developing solvent systems used include, but are not limited to, dichloromethane and methanol systems, n-hexane and ethyl acetate systems, and petroleum ether and ethyl acetate systems. The volume ratio of the solvents is adjusted according to the polarity of the compounds or by adding triethylamine.

[0267] Column chromatography generally uses Qingdao Ocean 200-300 mesh silica gel as the stationary phase. Eluent systems include, but are not limited to, dichloromethane and methanol systems and n-hexane and ethyl acetate systems. The volume ratio of the solvents is adjusted according to the polarity of the compound, and a small amount of triethylamine can also be added for adjustment.

[0268] Unless otherwise specified in the examples, the reaction temperature is room temperature (20°C to 30°C).

[0269] Unless otherwise specified, the reagents used in the examples were purchased from Acros Organics, Aldrich Chemical Company, Nanjing Yaoshi Technology, or Shanghai Shuya Pharmaceutical Technology.

[0270] The above embodiments do not limit the solutions of the present application in any way. In addition to those described herein, various modifications of the present invention will be apparent to those skilled in the art based on the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. Each reference cited in this application (including all patents, patent applications, journal articles, books and any other disclosures) is incorporated by reference in its entirety.

[0271] In conventional synthesis methods, preparation examples, embodiments and intermediate synthesis examples, the meanings of the abbreviations are shown in the following table.

[0272] Example

[0273] Example 1: Preparation of camptothecin compounds

[0274] Example 1.1: Preparation of Compound A1

[0275] Step 1: Compound A1-1 (6 g, 36.13 mmol) and compound A1-2 (6 g, 22.81 mmol) were placed in a 2 L three-necked flask under nitrogen. Glacial acetic acid (600 mL) was added to dissolve the mixture. Concentrated hydrochloric acid (120 mL) was then added and refluxed at 125°C for 16 hours. The reaction mixture was dried and the crude product was purified by column chromatography to obtain compound A1-3 (8.6 g, yield: 95.9%).

[0276] Step 2: Compound A1-3 (8.60 g, 21.87 mmol) was placed in a 500 mL three-necked flask and dissolved in glacial acetic acid (123 mL). Acetyl chloride (20.60 g, 262.52 mmol) was added under nitrogen and heated at 75°C for 1 hour. The reaction solution was spin-dried and the crude product was purified by column chromatography to obtain compound A1-4 (9.4 g, 98.7%).

[0277] Step 3: Compound A1-4 (9.4 g, 21.6 mmol) was placed in a 250 mL single-necked flask and dissolved in DCM (108 mL). The mixture was cooled to 0°C and m-CPBA (13.15 g, 64.81 mmol) was added. The mixture was returned to room temperature and allowed to react for 1 hour. Saturated aqueous sodium bicarbonate solution was added dropwise to the reaction solution to adjust the pH to approximately 8. The layers were separated and the organic phase was collected. The aqueous phase was extracted with DCM (80 mL x 3). The combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure and the resulting crude product was purified by column chromatography to afford compound A1-5 (7.5 g, yield: 78.9%).

[0278] Step 4: Compound A1-5 (3 g, 6.65 mmol) was placed in a 100 mL single-necked flask and dissolved in DMF (38 mL). The mixture was cooled to 0°C and slowly added dropwise with a 2 M solution of oxalyl chloride in DCM (8.3 mL). After the addition, the mixture was returned to room temperature and allowed to react for 2 hours. The reaction solution was poured into water (400 mL) and extracted with DCM (100 mL x 4). The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure, and the crude product was purified by column chromatography to afford compound A1-6 (2.4 g, yield: 77.1%).

[0279] Step 5: Compound A1-6 (300 mg, 0.84 mmol) was dissolved in 1,4-dioxane (6 mL), and isoamyl alcohol A1-7 (5 mL) and trifluoromethanesulfonic acid (287 mg, 1.9 mmol) were added. The reaction solution was purged with nitrogen three times and reacted at 60°C for 4 hours. LCMS monitoring of the reaction showed that the starting material was completely reacted. The reaction solution was directly concentrated, most of the organic solvent was removed by vortexing, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed three times with saturated sodium chloride, and dried over anhydrous sodium sulfate. After concentration, the mixture was dry-mixed and purified by silica gel column chromatography (100% Hep ~ 67% EA in EA) to obtain A1-8 (50 mg, yield: 15.0%). MS-ESI: m / z 522.2 = M+H +

[0280] Step 6: Dissolve compound A1-8 (50 mg, 0.096 mmol) in tetrahydrofuran (2 mL), add wet Pd / C (5%, 20 mg), and replace the reaction solution with hydrogen three times. Under hydrogen protection, react at room temperature 25°C for 6 hours. LCMS monitors the completion of the reaction. The reaction solution is directly filtered, and the filtrate is concentrated to dryness to obtain A1-9 (46 mg, yield: 97.9%), which is directly used in the next reaction. MS-ESI: m / z 492.2 = M+H +

[0281] Step 7: Compound A1-9 (48 mg, 0.10 mmol) was dissolved in anhydrous methanol (2 mL), and a 1 mol / L solution of MeONa (0.20 mL, 0.20 mmol) in methanol was added. The atmosphere was purged with nitrogen three times and the mixture was reacted at room temperature (25°C) for 2 hours. The reaction was monitored by LCMS. After cooling the reaction solution with ice water, the pH of the system was adjusted to 3-4 with 1 mol / L hydrochloric acid. The quenched reaction solution was directly purified using a preparative column (40% MeCN in H2O containing 0.1% FA) and lyophilized to obtain A1 (4.12 mg, yield: 5.21%).

[0282] MS-ESI: m / z 450.3=M+H +

[0283] 1 H NMR (400MHz, DMSO-d6) δ7.76(d,J=8.9Hz,1H),7.21(m,2H),7.14(s,1H),6.49(s,1H),5.85(s,2H),5.54(s,2H), 5.39(d,J=4.1Hz,2H),4.63(t,2H),1.94(m,1H),1.85(m,2H),1.76(m,2H),1.04–0.91(m,6H),0.89–0.84(m,3H).

[0284] Example 1.2: Preparation of Compound A12

[0285] Step 1: Dissolve compound A1-6 (400 mg, 0.85 mmol) in 1,4-dioxane (10 mL), add A12-1 (9 mL, 4.2 mmol) and trifluoromethanesulfonic acid (287 mg, 1.9 mmol), replace the reaction solution with nitrogen three times, and place it at 60°C to react for 4 hours. LCMS monitoring reaction shows that the raw material is completely reacted. The reaction solution is directly concentrated, most of the organic solvent is removed by rotation, extracted with ethyl acetate three times, washed with saturated sodium chloride three times, and dried over anhydrous sodium sulfate. After concentration, dryness is mixed and purified by silica gel column chromatography (100% DCM ~ 10% MeOH in DCM) to obtain A12-2 (109 mg, yield: 23.8%). MS-ESI: m / z 538.2M+H +

[0286] Step 2: Compound A12-2 (109 mg, 0.20 mmol) was dissolved in tetrahydrofuran (6 mL), 10% wet Pd / C (60 mg) was added, and the reaction solution was replaced with hydrogen three times. Under hydrogen protection, the reaction was carried out at room temperature of 25°C for 6 hours. The reaction was monitored by LCMS. The reaction solution was directly filtered, and the filtrate was concentrated to dryness to obtain A12-3 (87 mg, yield: 84.5%), which was directly used in the next reaction. MS-ESI: m / z 508.1M+H +

[0287] Step 3: Compound A12-3 (87 mg, 0.17 mmol) was dissolved in anhydrous methanol (4 mL), and a 1 mol / L methanol solution of MeONa (0.35 mL, 0.35 mmol) was added. The reaction solution was purged with nitrogen three times and reacted at room temperature (25°C) for 2 hours. After cooling the reaction solution with ice water, the pH of the system was adjusted to 3-4 with 1 mol / L hydrochloric acid. The quenched reaction solution was directly purified using a preparative column (40% MeCN in H2O containing 0.1% TFA) and lyophilized to obtain A12 (4.12 mg, yield: 5.21%) as a yellow solid.

[0288] MS-ESI: m / z 466.1M+H +

[0289] 1H NMR (400MHz, DMSO) δ7.77(d,J=9.0Hz,1H),7.25–7.11(m,3H),6.46(s,1H),5.89(s,2H),5.54(s,2H),5. 40(s,2H),4.69(m,2H),3.83(t,2H),3.73–3.68(m,1H),1.86(m,2H),1.15(d,J=6.1Hz,6H),0.87(t,3H).

[0290] The following compounds were synthesized by referring to the methods of Examples 1.1 and 1.2 using appropriate starting materials.

[0291] Example 1.3: Preparation of Compound B-4

[0292] Step 1: Compound A1-2 (3 g, 11.39 mmol) and compound B4-2 (2.5 g, 18.22 mmol) were placed in a 1 L three-necked flask under nitrogen. Glacial acetic acid (300 mL) was added to dissolve the mixture, followed by concentrated hydrochloric acid (60 mL). The mixture was refluxed at 125°C for 16 hours. The reaction mixture was dried and the crude product was purified by column chromatography to obtain compound B4-3 (3.8 g, yield: 91.1%). MS-ESI: m / z 367.1 = M+H +

[0293] Step 2: AcCl (3.21 g, 40.95 mmol) was added dropwise to a solution of B4-3 (3 g, 8.19 mmol) in AcOH (30 mL) and heated to 75°C for 4 hours. The reaction mixture was poured into ice water (30 mL) and extracted with DCM (20 mL*3). The organic phases were combined, dried over anhydrous Na2SO4, and concentrated to give a crude product, which was purified by column chromatography (0-30% dichloromethane / EtOAc) to give compound B4-4 (1.8 g, yield: 53.8%). MS-ESI: m / z 409.1 = M+H +

[0294] Step 3: Compound B4-4 (1.6 g, 3.92 mmol) was dissolved in DCM (30 mL), cooled to 0°C, and m-CPBA (2.03 g, 11.76 mmol) was added. The mixture was stirred at room temperature overnight. The reaction mixture was washed with saturated Na2S2O5 (50 mL) and saturated NaHCO3 (50 mL). The organic phase was concentrated to afford compound B4-5 (1.1 g, yield: 66.1%).

[0295] Step 4: Compound B4-5 (1.0 g, 2.36 mmol) was dissolved in DMF (200 mL), cooled to 0°C, and (COCl) (0.75 g, 5.91 mmol) was added. The mixture was reacted at room temperature for 2 hours. The reaction solution was poured into water (500 mL) and extracted with EA (300 mL*4). The organic phases were combined, dried over anhydrous Na2SO4, and concentrated to give a crude product, which was purified by column chromatography (0-50% ethyl acetate / EtOAc) to give compound B4-6 (0.77 g, 73.7%).

[0296] Step 5: Dissolve B4-6 (600 mg, 1.35 mmol) in ethylene glycol (10 mL) and add TfOH (2 mL). Stir the reaction at 50°C for 48 hours. LCMS analysis indicates that the reaction is complete. Pour the reaction solution into water, and a brown solid precipitates. The filter cake is collected by filtration to obtain 480 mg of compound B4-7. The crude product is not included in the yield. MS-ESI: m / z 469.1 = M+H +

[0297] Step 6: Compound B4-7 (200 mg, 0.43 mmol), methanol (10 mL), and K2CO3 (84 mg) in water (2 mL) were added to a single-necked flask and stirred at room temperature for 3 hours. The pH was then adjusted to 3-4 with 0.5 M dilute hydrochloric acid and stirred for 10 minutes. The reaction solution was extracted three times with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by preparative HPLC to obtain B4 (20 mg, yield: 10.9%) as a yellow solid.

[0298] MS-ESI: m / z 427.1=M+H +

[0299] 1 H NMR (400MHz, DMSO-d6) δ7.43(t,J=8.1Hz,1H),7.25(s,1H),7.17(d,J=8.1Hz,1H),6.70(d,J=7.6Hz,1H),6.51(s, 2H),5.54(s,2H),5.42(s,2H),4.67(t,J=4.4Hz,2H),3.90–3.85(m,2H),1.92–1.80(m,2H),0.87(t,J=7.3Hz,3H).

[0300] The following compounds were synthesized by referring to the method of Example 1.3 using appropriate starting materials.

[0301] Example 2: Preparation of linker intermediates

[0302] Example 2.1: Preparation of intermediate Int9

[0303] Compound Int9-1 (500 mg, 1.42 mmol) was dissolved in DMF (5 ml), and HATU (810 mg, 2.13 mmol) and DIEA (549 mg, 4.26 mmol) were added. After stirring at room temperature for 30 minutes, compound Int9-2 (369 mg, 1.42 mmol) was added and the reaction continued for 2 hours. After completion of the reaction, ethyl acetate was added, and the pH was adjusted to approximately 6 with 2M citric acid under stirring. The layers were separated, and the organic phase was washed twice with saturated brine, dried, concentrated, and purified by reverse-phase HPLC to obtain compound Int9 (422 mg, 50% yield).

[0304] The following intermediates were synthesized by referring to the method of Example 2.1:

[0305] Example 3: Preparation of linker-payload for ligand drug conjugates

[0306] Preparation of compound AA1

[0307] Compound Int2 (100 mg, 0.28 mmol), compound A1 (125 mg, 0.28 mmol), HATU (159 mg, 0.42 mmol), and DIPEA (108 mg, 0.84 mmol) were dissolved in DMF (5 mL) and stirred at room temperature for 12 hours. The mixture was quenched with water and extracted with ethyl acetate. The organic phase was separated and dried over anhydrous sodium sulfate and concentrated to obtain the crude product, which was purified by preparative HPLC to obtain compound AA1 (110 mg, 50% yield). MS: [M+H] + , 785.3.

[0308] Preparation of compound AA18

[0309] Step 1: To a DMF (3 mL) solution of compound A12 (50 mg, 0.1 mmol), compound int17 (97.5 mg, 0.2 mmol), HATU (82 mg, 0.2 mmol), and HOAT (30 mg, 0.2 mmol) was added TMP (20 mg, 0.4 mmol). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was spin-dried and purified by reverse phase column chromatography (H2O:Acetonitrile = 0-100%) to obtain AA18-1 as a yellow solid (60 mg, yield: 62.5%). LCMS: [M+H] = 901.3

[0310] Step 2: To a solution of compound AA18-1 (30 mg, 0.033 mmol) in THF (1 mL) was added 6 M HCl (1 mL). The reaction mixture was stirred at room temperature for 48 hours. The reaction mixture was spin-dried and purified by reverse-phase column chromatography (H₂O:Acetonitrile = 0-100%) to afford AA18 (4 mg, yield: 14.2%) as a white solid.

[0311] LCMS: [M+1] + =845.2

[0312] 1 H NMR (400MHz, DMSO) δ12.15(s,1H),10.21(s,1H),8.70(s,1H),8.37(t,J=5.7Hz,1H),8.18(s,1H),8.06–7.95(m,2H) ,7.28(s,1H),6.98(s,2H),6.52(s,1H),5.63(s,2H),5.43(s,2H),4.86–4.67(m,2H),4.23(dd,J=13.8,7.5Hz,1H), 3.95(s,2H),3.89–3.81(m,2H),3.73(dt,J=12.2,6.2Hz,1H),2.36–2.24(m,2H),2.22–2.08(m,2H),2.02–1.72(m,4 H), 1.48 (ddd, J = 21.9, 14.9, 7.4Hz, 4H), 1.21 (dd, J = 15.2, 8.2Hz, 2H), 1.13 (d, J = 6.1Hz, 6H), 0.88 (t, J = 7.3Hz, 4H).

[0313] Example 4: Preparation of Ligand Drug Conjugates

[0314] Antibodies serving as ligands are prepared according to conventional methods. For example, vector construction can be performed, followed by transfection into eukaryotic cells such as HEK293 or CHO cells, followed by purification and expression. Ligand-drug conjugates were prepared using the anti-CEACAM5 antibody Tusamitamab (prepared with reference to WO2014079886A1), the anti-HER3 antibody Patritumab (prepared with reference to WO2007077028A2), and the anti-GPC3 antibody Codrituzumab (prepared with reference to WO2006006693) as examples.

[0315] Anti-HER3 antibody Patritumab heavy chain amino acid sequence

[0316] Anti-HER3 antibody Patritumab light chain amino acid sequence

[0317] Anti-GPC3 antibody Codrituzumab heavy chain amino acid sequence

[0318] Anti-GPC3 antibody Codrituzumab light chain amino acid sequence

[0319] Anti-CEACAM5 antibody Tusamitamab heavy chain amino acid sequence

[0320] Anti-CEACAM5 antibody Tusamitamab light chain amino acid sequence

[0321] Example 4.1: Preparation of ADC-1A

[0322] At 37°C, prepared tris(2-carbonylethyl)phosphine hydrochloride (10 mM, 0.135 mL, 1.35 μmol) was added to the buffer of the antibody Tusamitamab (14.0 mM succinic acid-sodium hydroxide + 108 mM NaCl pH 6.0; 20 mg, 10.0 mg / mL, 0.135 μmol), and the mixture was placed in a water bath shaker and shaken at 37°C for 3 hours. After stopping the reaction, the excess TCEP was removed by ultrafiltration using 14.0 mM succinic acid-sodium hydroxide + 108 mM NaCl pH 6.0 buffer;

[0323] Compound AA1 (1.29 mg, 1.65 μmol) was dissolved in 0.13 mL of DMSO and added to the above solution. The mixture was placed in a water bath shaker and shaken at 22° C. for 2 hours to stop the reaction. The reaction solution was desalted and purified using a Sephadex G25 gel column (elution phase: 20 mM histidine-hydrochloric acid pH 5.5) to obtain a solution of the exemplary product ADC-1A (20 mM histidine-hydrochloric acid pH 5.5; 18.4 mg, 4.96 mg / mL, yield: 92%), which was stored at 4° C.

[0324] The DAR value q was calculated by LC-MS analysis and detection = 7.82.

[0325] Example 4.2: Preparation of ADC-18A

[0326] At 37°C, prepared tris(2-carbonylethyl)phosphine hydrochloride (10 mM, 0.135 mL, 1.35 μmol) was added to the buffer of the antibody Tusamitamab (14.0 mM succinic acid-sodium hydroxide + 108 mM NaCl pH 6.0; 20 mg, 10.0 mg / mL, 0.135 μmol), and the mixture was placed in a water bath shaker and shaken at 37°C for 3 hours. After stopping the reaction, the excess TCEP was removed by ultrafiltration using 14.0 mM succinic acid-sodium hydroxide + 108 mM NaCl pH 6.0 buffer;

[0327] Compound AA18 (1.39 mg, 1.65 μmol) was dissolved in 0.14 mL of DMSO and added to the above solution. The mixture was placed in a water bath shaker and shaken at 22° C. for 2 hours to stop the reaction. The reaction solution was desalted and purified using a Sephadex G25 gel column (elution phase: 20 mM histidine-hydrochloric acid pH 5.5) to obtain a solution of the exemplary product ADC-18A (20 mM histidine-hydrochloric acid pH 5.5; 17.2 mg, 4.26 mg / mL, yield: 86%), which was stored at 4° C.

[0328] The DAR value q was calculated by LC-MS analysis and detection = 7.91.

[0329] Example 5: In vitro tumor cell proliferation inhibition test of compounds

[0330] Purpose of the test

[0331] In order to detect the inhibitory activity of drug compounds on the proliferation of NCI-N87 cells, JIMT-1 and MBA-MB-231 tumor cells in vitro, cells were treated with different concentrations of compounds in vitro and cultured for 6 days. Luminescent Cell Viability Assay (Promega, Catalog No. G7558) was used to detect cell proliferation. 50 The in vitro activity of the compound was evaluated.

[0332] Experimental methods

[0333] The following uses the in vitro proliferation inhibition test method of NCI-N87 cells as an example to illustrate the method for testing the in vitro proliferation inhibition activity of the compounds of the present application on tumor cells. This method is also applicable to, but not limited to, testing the in vitro proliferation inhibition activity of other tumor cells.

[0334] 1. Cell culture: NCI-N87 cells were cultured in RPMI-1640 medium supplemented with 10% FBS.

[0335] 2. Cell preparation: Take NCI-N87 cells in the logarithmic growth phase, wash once with PBS, add 2-3 ml of trypsin to digest for 2-3 minutes, wait until the cells are completely digested, add 10-15 ml of cell culture medium to elute the digested cells, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, and then add 10-20 ml of cell culture medium to resuspend the cells to make a single-cell suspension.

[0336] 3. Cell plating: Mix the NCI-N87 single cell suspension and adjust the viable cell density to 6×10 4 After adjusting the density, the cell suspension was mixed and added to a 96-well cell culture plate at 50 μl / well. The culture plate was cultured in an incubator for 18 hours (37°C, 5% CO2).

[0337] 4. Compound Preparation: Dissolve the compound in DMSO and prepare a stock solution with an initial concentration of 10 mM. Eight concentrations of small molecule compounds are used: 300, 100, 30, 10, 3, 1, 0.3, and 0.1 nM.

[0338] 5. Sample addition: Add different concentrations of the test sample to the culture plate, with two replicates for each sample. Incubate the culture plate in an incubator for 6 days (37°C, 5% CO2).

[0339] 6. Color development: Take out the 96-well cell culture plate, add 50ul CTG reagent to each well, and incubate at room temperature for 10 minutes.

[0340] 7. Plate reading operation: Take out the 96-well cell culture plate, place it in a microplate reader, and measure the chemiluminescence using the microplate reader.

[0341] Data Analysis

[0342] The data were processed and analyzed using Microsoft Excel and Graphpad Prism 5.

[0343] Table 3. IC values ​​of the small molecule fragments in this application for inhibition of NCI-N87 cell proliferation in vitro 50 value.

[0344] Conclusion: According to the results in Table 3, the small molecule fragments in this application have obvious proliferation inhibitory activity on NCI-N87 cells, JIMT-1 cells, and MDA-MB-231 cells.

[0345] Example 6: In vitro cell proliferation inhibition activity test of antibody-drug conjugates

[0346] Implementation: 6.1: In vitro proliferation inhibition activity test of NCI-N87 / JIMT-1 cells

[0347] use The chemiluminescent cell viability assay (CTG method) was used to evaluate the inhibitory effect of the anti-Her2 antibody trastuzumab (ADC) coupled with a camptothecin compound on cell proliferation in Her2-positive human gastric cancer cells NCI-N87 and human breast cancer cells JIMT-1 after incubation for 6 days.

[0348] Cells in the logarithmic growth phase were collected and plated in a 96-well cell culture plate at a density of 6,000 cells / well. The cell plates were placed in a 37°C, 5% CO2 incubator for overnight culture. On the second day of the experiment, the ADC drug of the camptothecin compound was diluted 3-fold with complete culture medium to obtain 9 concentration gradients (starting with the highest concentration of 300nM). After the drug was added, 100μL / well was added to the cell culture plate. The complete culture medium was used as a blank control, and 3 replicates were set up; the plate was continued to be incubated in a 37°C, 5% CO2 incubator for 6 days. After the incubation was completed, the cell culture plate was removed and equilibrated to room temperature. 50μL CTG detection reagent (Promega, Cat#: G7573) was added to each well. After shaking and mixing, it was placed in the dark for 10 minutes, and the signal value was read using an enzyme-linked microplate reader. GraphPad Prism software was used to draw a S-shaped dose-response curve using a nonlinear regression model and calculate the IC 50 Cell viability calculation formula = (Lum 待测药 -Lum 空白对照 ) / (Lum 溶剂空白对照 -Lum 空白对照 )×100%.

[0349] Experimental conclusion: The antibody-drug conjugate of the present application has significant proliferation inhibitory activity on Her2-positive human gastric cancer cell NCI-N87 and human breast cancer JIMT-1 cells.

[0350] Example 7: In vivo tumor inhibition test of antibody drug conjugates

[0351] To evaluate the inhibitory effect of the ADC drug of the present invention on tumor formation in vivo, after forming transplanted tumors in mice using Her2-positive human breast cancer cells JIMT-1, the anti-tumor effect of the ADC drug of the present invention was evaluated.

[0352] Evaluation of the efficacy of antibody-drug conjugates in JIMT-1 human breast cancer cell-bearing mice

[0353] 1. Test drugs and materials

[0354] Blank control group (control group): normal saline

[0355] ADC (treatment group): 5 mg / kg, single dose

[0356] 2. Preparation method: All samples were diluted with physiological saline.

[0357] 3. Experimental animals: 8-week-old female BALB / c-nude mice were purchased from Jicui Yaokang Biotechnology Co., Ltd.

[0358] 4. Test method:

[0359] 1×10 7 JIMT-1 cells were inoculated subcutaneously at the right anterior scapula of 8-week-old female BALB / c-nude mice. 3 Tumor-bearing mice were randomly divided into groups using StudyDirect™. Starting on day 0, the ADC of the present invention was injected intravenously (iv) once every 7 days for a total of two injections at a dose of 5 mg / kg. Tumor volume and body weight were measured twice weekly and the data were recorded.

[0360] Each vehicle control group or treatment group consisted of 5 mice. Tumor inhibition rate (TGI%) was calculated by measuring tumor volume. Tumor inhibition rate (TGI%) = 100% - (tumor volume of treatment group on the day of measurement - tumor volume of treatment group on day 0) / (tumor volume of control group on the day of measurement - tumor volume of control group on day 0).

[0361] The experimental results show that the antibody-drug conjugate of the present invention exhibits significant tumor-suppressing activity after a single administration.

[0362] Example 8: Plasma stability test of antibody drug conjugates

[0363] To evaluate the plasma stability of the antibody drug conjugates of the present invention, the antibody drug conjugates of the present invention were incubated in human, rat, and monkey plasma for 21 days. Samples were taken at 0 hours, 8 hours, 1 day, 4 days, 7 days, 14 days, and 21 days to detect the drug-ligand coupling rate (DAR value) and free drug.

[0364] Incubation of Antibody Drug Conjugates in Plasma

[0365] The antibody drug conjugate of the present invention was diluted with plasma to a final concentration of 150 μg / mL and incubated at 37°C in the dark for 21 days. Samples were collected at T0 (the sample was immediately frozen to -70°C after dilution within 30 minutes), 2 hours, 8 hours, day 1 (24 hours), day 4, day 7, day 14, and day 21. All samples were stored in a -70°C freezer before analysis. Only the DAR values ​​were analyzed for samples collected at 2 hours and 8 hours.

[0366] Analysis of free drugs by LC-MS / MS

[0367] Protein precipitation

[0368] Plasma samples were thawed, and 291.9 μL was collected for free drug analysis. A precipitant (400 μL of 0.1% formic acid, 200 ng / mL tolbutamide, and 200 ng / mL labetalol in acetonitrile) was added to each sample and mixed thoroughly. The mixture was shaken for at least 20 minutes to precipitate plasma proteins. The samples were centrifuged at 4°C, 4000 rpm, and 150 μL of the supernatant was collected for LC-MS / MS analysis.

[0369] The equipment used for free drug analysis is shown in the table below:

[0370] The LC and MS parameters are as follows:

[0371] LC-MS determination of drug-ligand coupling rate

[0372] Immobilized magnetic beads capture antibody drug conjugates

[0373] 25 μL of streptavidin magnetic beads were added to each well of a 96-well plate, and the stock buffer was discarded. The beads were eluted with 200 μL of PBS buffer and treated with 80 μL of biotinylated antigen protein and another 100 μL of PBS buffer at room temperature for 120 minutes. After complete immobilization of the antigen protein, the beads were eluted twice with 200 μL of PBS buffer. 20 μL of antibody-drug conjugate plasma sample and 180 μL of PBS buffer were added to each well, followed by shaking at room temperature for 120 minutes to ensure complete capture of the antibody-drug conjugate in the plasma, and the supernatant was removed. The beads were washed twice with PBS buffer, followed by the addition of 50 μL of elution buffer (1% formic acid in water) and treatment at room temperature for 20 minutes. Subsequently, 5 μL of neutralization buffer (1 M NH4HCO3, pH 8.5) and 5 μL of DTT (1 M) were added, and the captured antibody-drug conjugate was reduced at room temperature for 60 minutes before analysis by LC-MS.

[0374] Drug-ligand coupling rate analysis

[0375] The LC-MS parameters are as follows:

[0376] The results showed that after 21 days of incubation in plasma, no drug or only a very small amount of drug was shed from the antibody-drug conjugate of the present invention, and there was no significant change in the drug-ligand coupling rate. The antibody-drug conjugate of the present invention has extremely high plasma stability, indicating that the antibody-drug conjugate of the present invention has better safety.

[0377] Example 9: Antibody Drug Conjugate Stock Solution (Liquid Preparation) Stability

[0378] To evaluate the stability of the stock solution of the antibody-drug conjugate of the present invention, the ADC stock solution of the present invention was concentrated to 20 mg / mL, dispensed into 2 mL cryovials, and incubated in a 40°C incubator. Samples were taken at 0 hour, 1 week, 2 weeks, 1 month, and 2 months for concentration, drug-ligand coupling rate (DAR value), SEC purity and aggregation, CE-SDS (non-reducing) purity, CE-SDS (reducing) purity, charge heterogeneity, and free drug detection. At the same time, the appearance of the liquid was visually observed for clarity and the formation of precipitation.

[0379] The sample concentration was detected by UV method.

[0380] The drug-ligand coupling rate (DAR value) was detected by hydrophobic HPLC (HIC) or reverse phase HPLC (RP-HPLC).

[0381] The drug-ligand coupling rate (DAR value) was detected by LC-MS.

[0382] Reverse phase HPLC (RP-HPLC) was used to detect free drug.

[0383] The SEC purity and aggregation of the samples were determined by SEC-HPLC (TOSOH G3000 SW SEC column).

[0384] Maurice was used to detect the purity of CE-SDS (non-reduced) and CE-SDS (reduced).

[0385] Imaging capillary isoelectric focusing (iCIEF) was used to detect charge heterogeneity.

[0386] The results showed that after incubation at 40°C for one week to two months, the antibody-drug conjugate stock solution of the present invention showed no significant changes in concentration, drug-ligand coupling ratio (DAR), SEC purity and aggregation, CE-SDS (non-reducing) purity, CE-SDS (reducing) purity, charge heterogeneity, and free drug. The antibody-drug conjugate stock solution of the present invention has extremely high stability.

[0387] Example 10: Pharmacokinetic and toxicological studies of single or multiple doses in monkeys

[0388] After a single or multiple intravenous injections of the antibody drug conjugate of the present invention are given to monkeys, the pharmacokinetic properties of the drug in the monkeys are investigated, and the toxicity of the animals is observed.

[0389] Test methods

[0390] Pharmacokinetics: After a single intravenous infusion of different doses of the antibody-drug conjugate of the present invention into monkeys, blood samples were collected at multiple time points and the concentration of the drug in the blood was measured using an appropriate specific detection method.

[0391] Toxicology studies: After single or multiple intravenous infusions of different doses of the antibody-drug conjugate of the invention in monkeys, the animals' tolerance to the antibody-drug conjugate of the invention, as well as drug-related toxicity, were assessed through clinical observation, body weight and food intake, hematology, blood biochemistry, urine, gross anatomy, and histopathology.

[0392] Results showed that after single or multiple intravenous infusions of the antibody-drug conjugate of this invention in monkeys, the pharmacokinetic properties of the total antibody and ADC were similar, with very low free toxin concentrations, demonstrating that the antibody-drug conjugate of this invention has high stability and good pharmacokinetic properties in vivo. The animals were well tolerated, with no severe or intolerable drug-related toxicities, demonstrating that the safety of the antibody-drug conjugate of this invention is controllable.

[0393] Sequences of this application (Kabat numbering scheme):

Claims

1. A ligand-drug conjugate, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein the ligand-drug conjugate comprises a ligand and a structure represented by formula (Ia) or formula (Ib): in, The wavy line indicates direct or indirect connection to the ligand through -NH- or X; R 1a Selected from hydrogen, halogen, cyano, amino, hydroxyl, -C 1-6 Alkyl and -C 1-6 alkoxy; Y is selected from -C 1-6 Alkylene, -C 2-6 Alkenylene, 3-10 membered cycloalkylene and 3-10 membered heterocyclylene; each of the alkylene, alkenylene, cycloalkylene and heterocyclylene is optionally substituted by one or more selected from hydrogen, halogen, hydroxyl, oxo, -C 1-6 Alkyl and -C 1-6 Substitution of alkoxy groups; Z is absent or selected from -O-, -S-, -N(R 3a )-、-(C=O)-、-S(O)2-、-C 1-6 Alkylene, -OC 1-6 Alkylene, 3-10 membered cycloalkylene and 3-10 membered heterocyclylene; each of the alkylene, cycloalkylene and heterocyclylene is optionally substituted by one or more selected from hydrogen, halogen, hydroxyl, oxo, -C 1-6 Alkyl and -C 1-6 Substitution of alkoxy groups; R 3a Selected from hydrogen, hydroxyl, -C 1-6 Alkyl, -C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, -(C=O)-C 1-6 Alkyl and -S(O)2-C 1-6 Alkyl; the alkyl, cycloalkyl and heterocycloalkyl groups are each optionally substituted by one or more radicals selected from hydrogen, halogen, hydroxy, oxo, -C 1-6 Alkyl and -C 1-6 Substitution of alkoxy groups; R 2a Selected from hydrogen, hydroxy, amino, cyano, -C 1-6 Alkyl, -C 1-6 Alkoxy, -C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl; the alkyl, alkoxy, cycloalkyl and heterocycloalkyl are each optionally substituted by one or more selected from hydrogen, halogen, hydroxy, oxo, -C 1-6 Alkyl and -C 1-6 Substitution of alkoxy groups; X is selected from: -O- and -N(R 3b-2 )-; R 1b 、R 2b are independently selected from hydrogen, halogen, hydroxy, nitro, -C 1-6 Alkyl and -C 1-6 alkoxy; or R 1b With R 2b and the atoms to which they are attached together form a 4-6 membered carbocyclic ring or a 4-6 membered heterocyclic ring; t is an integer selected from 1 to 10; R 3b-1 Each independently selected from hydrogen, halogen, hydroxy, amino, cyano, nitro, -C 1-6 Alkyl and -halogenated C 1-6 alkyl; Or, any two R 3b-1 and the atoms to which it is attached together form an oxo group, a 3-6 membered carbocyclic ring or a 3-6 membered heterocyclic ring; R 3b-2 Selected from hydrogen, hydroxyl, -C 1-6 Alkyl, -C 3-6 Cycloalkyl and 3 to 6 membered heterocycloalkyl; said alkyl, cycloalkyl and heterocycloalkyl are each optionally substituted by one or more radicals selected from hydrogen, halogen, hydroxy, amino and -C 1-6 The alkyl group is substituted with a substituent.

2. The ligand-drug conjugate according to claim 1, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts or solvates thereof, wherein the ligand-drug conjugate comprises a ligand and a structure represented by formula (IIa) or formula (IIb): in, The wavy line indicates the L 1a linked to the ligand; L 1a Selected from: L 2 -(C(R L21 )2) n -; Wherein, n is a natural number from 0 to 50, such as 0, 1, 2, 3, 4, 5, 6, 7 or 8; L 2 Any-C(R L21 )2- units are each optionally replaced by the following structural units: -Cy-, -C(O)-, -NR L22 -, -O-, -S-, -SO-, -SO2-, -P(R L22 )-、-P(=O)(R L22 )-、-C(=S)-、-C(=NR L22 )-, -N=N-, -C=N-, -N=C-, -Cy- is selected from phenylene, 5 to 8 membered heteroarylene, 3 to 10 membered heterocyclylene and 3 to 10 membered cycloalkylene, wherein said -Cy- is unsubstituted or independently substituted by one or more R cx replace; R L21 、R L22 、R cx Each independently selected from hydrogen, deuterium, halogen, -NO2, -CN, -OR L2a 、-SR L2a 、-N(R L2a )2, -N + (R L2a )3, -C(O)R L2a 、-CO2R L2a 、-C(O)C(O)R L2a 、-C(O)CH2C(O)R L2a 、-S(O)R L2a 、-S(O)2R L2a 、-C(O)N(R L2a )2、-SO2N(R L2a )2、-OC(O)R L2a 、-N(R L2a )SO2R L2b 、-N(R L2a )COR L2b 、-(CH2) y -CO-(N(Me)CH2C(O)) m -OR L2a 、-(CH2) y -CO-(N(Me)CH2C(O)) m -NHR L2a 、-(CH2) y -CO-(N(Me)CH2C(O)) m -N + (R L2a )3, -(CH2) y -NHCOCH2(OCH2CH2)OR L2a 、-(CH2) y -NH(COCH2(N(Me)) m -R L2a 、-(CH2) y -CONH-(CH2CH2O) m -R L2a 、-(CH2) y -NHCO-(CH2CH2O) m -R L2a 、-(CH2CH2O) m -R L2a 、-(COCH2N(Me)) m -R L2a 、-COCH2(OCH2CH2) m -OR L2a 、-CO-(CH2CH2O) m -R L2a 、-CO-(CH2) y -CONH-(CH2CH2O) m -R L2a 、-CO-(CH2) y -NHCO-(CH2CH2O) m -R L2a and R L2a Optionally substituted -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, 3-8 membered cycloalkyl, 4-10 membered heterocycloalkyl, 6-10 membered aryl, and 3-10 membered heteroaryl; m and y are natural numbers from 0 to 50, such as 0, 1, 2, 3, 4, 5, 6, 7, or 8; R L2a 、R L2b Each independently selected from hydrogen, deuterium, halogen, -NO2, -CN, -OH, -SH, -NH2, -N(Me)2, -CO2H, -S(O)2Me, -S(O)2OH, -C(O)NH2, -SO2NH2, -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, 3-8 membered cycloalkyl, 4-10 membered heterocycloalkyl, 6-10 membered aryl or 3-10 membered heteroaryl; L 3 Does not exist or is an amino acid residue, a short peptide consisting of 2-10 amino acid residues, or any combination of the above groups, wherein the amino acid residue is a natural amino acid residue or a non-natural amino acid residue; Tr does not exist or is or any combination of the above groups; R Tr 、R Tr1 and R Tr2 Each independently selected from hydrogen, deuterium, halogen, -NO2, -CN, -OH, -SH, -NH2, -CO2H, -S(O)2OH, -C(O)NH2, -SO2NH2, -OC(O)NH2, -CH2CO-(N(Me)CH2C(O)) z -OR Tra 、-CH2CO-(N(Me)CH2C(O)) z -NHR Tra 、-(CH2CH2O) z -R Tra 、-CONH-(CH2CH2O) z -R Tra Or by R Tra Optionally substituted -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, 3-8 membered cycloalkyl, 4-10 membered heterocycloalkyl, 6-10 membered aryl or 3-10 membered heteroaryl; R Tra For hydrogen, deuterium, halogen, -NO2, -CN, -OH, -SH, -NH2, -N(Me)2, -S(O)2Me, -CO2H, -S(O)2OH, -C(O)NH2, -SO2NH2, -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 alkynyl, 3-8 membered cycloalkyl, 4-10 membered heterocycloalkyl, 6-10 membered aryl or 3-10 membered heteroaryl, z is independently a natural number from 0 to 50; for example, 0, 1, 2, 3, 4, 5, 6, 7 or 8; X, Y, Z, R 1a 、R 2a 、R 1b 、R 2b 、R 3b-1 and t as defined in claim 1.

3. The ligand-drug conjugate according to claim 2, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts or solvates thereof, wherein the ligand-drug conjugate has a structure represented by Formula (IIIa) or Formula (IIIb): in: Ab is the ligand; q is the drug loading, which is an integer or decimal from 1 to 32, preferably an integer or decimal from 1 to 16; R 1a , X, Y, Z, R 2a 、R 1b 、R 2b 、R 3b-1 and t as defined in claim 1; and And L 1a 、L 2 、L 3 and Tr as defined in claim 2.

4. The ligand-drug conjugate according to any one of claims 1 to 3, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts or solvates, wherein R 1a Selected from hydrogen, halogen, -C 1-6 Alkyl; preferably, R 1a is selected from hydrogen, halogen; further preferably, R 1a Selected from hydrogen.

5. The ligand-drug conjugate according to any one of claims 1 to 4, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts or solvates thereof, wherein Y is selected from -C 1-6 Alkylene, 3-6 membered cycloalkylene; preferably, Y is selected from -C 1-6 Alkylene.

6. The ligand-drug conjugate according to any one of claims 1 to 5, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts or solvates thereof, wherein Z is absent or selected from -O-, -N(R 3a )-、-S(O)2-、-C 1-6 Alkylene; preferably, Z is absent or selected from -O-.

7. The ligand-drug conjugate according to any one of claims 1 to 6, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts or solvates, wherein R 3a Selected from hydrogen, -(C=O)-C 1-6 Alkyl, -S(O)2-C 1-6 Alkyl; preferably, R 3a Selected from -(C=O)-CH3 and -S(O)2-CH3.

8. The ligand-drug conjugate according to any one of claims 1 to 7, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts or solvates, wherein R 2a Selected from hydrogen, -C 1-6 Alkyl and -C 1-6 preferably, R 2a Selected from hydrogen and -C 1-6 alkyl.

9. The ligand-drug conjugate according to any one of claims 1 to 8, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts or solvates, wherein R 1b 、R 2b are each independently selected from hydrogen, halogen, -C 1-6 Alkoxy and -C 1-6 Alkyl; further preferably, R 1b 、R 2b are each independently selected from hydrogen, F, Cl and methyl.

10. The ligand-drug conjugate according to any one of claims 1 to 8, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts or solvates, wherein: R 1b With R 2b and the atoms to which it is attached together form a 5-membered heterocyclic ring; preferably, the 5-membered heterocyclic ring is 11. The ligand-drug conjugate according to any one of claims 1 to 10, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts or solvates, wherein: R 3b-1 are each independently selected from hydrogen, halogen, -C 1-6 Alkyl; preferably, R 3b-1 are each independently selected from hydrogen and -C 1-6 Alkyl; further preferably, R 3b-1 are each independently selected from hydrogen and methyl.

12. The ligand-drug conjugate according to any one of claims 1 to 11, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts or solvates, wherein: t is selected from an integer of 2-5; preferably, t is selected from 2 and 3.

13. The ligand-drug conjugate according to any one of claims 1 to 12, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts or solvates, wherein: R 3b-2 Selected from hydrogen and -C 1-6 Alkyl; preferably, R 3b-2 Selected from hydrogen.

14. The ligand-drug conjugate according to any one of claims 1 to 13, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: R 1a is selected from hydrogen, fluorine and chlorine; Y is selected from -C 1-6 Alkylene, cyclopropylene, cyclobutylene; Z is absent or selected from -O-, -N(R 3a )-、-S(O)2-、-C 1-6 alkylene; R 3a Selected from hydrogen, -(C=O)-C 1-6 Alkyl, -S(O)2-C 1-6 Alkyl; preferably, R 3a Selected from -(C=O)-CH3 and -S(O)2-CH3; R 2a Selected from hydrogen, -C 1-6 Alkyl and -C 1-6 alkyl halide; R 1b 、R 2b are each independently selected from hydrogen, F, Cl and methyl; Or, R 1b With R 2b and the atoms to which it is attached together form a 5-membered heterocyclic ring; preferably, the 5-membered heterocyclic ring is t is selected from 2 and 3; R 3b-1 are each independently selected from hydrogen and -C 1-6 alkyl; X is selected from: -O- and -N(R 3b-2 )-; R 3b-2 Selected from hydrogen and -C 1-6 alkyl.

15. The ligand-drug conjugate according to any one of claims 1 and 3 to 14, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: The structures represented by formula (Ia) and formula (Ib) are any of the following structures:

16. The ligand-drug conjugate according to any one of claims 2 to 14, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts or solvates, wherein L 1a Selected from: Preferably, L 1a Selected from 17. The ligand-drug conjugate according to any one of claims 2 to 14 and 16, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts or solvates, wherein L 2 -(CHR L21 ) n -; in, n is a natural integer selected from 0 to 50; for example, 0, 1, 2, 3, 4, 5, 6, 7 or 8; L 2 Any CHR in L21 The units are each optionally replaced by the following structural units: -Cy-, -C(O)-, -NR L22 -、-O-、 -Cy- is selected from phenylene, 5- to 6-membered heteroarylene, 4- to 10-membered heterocyclylene, and 3- to 6-membered cycloalkylene, wherein said -Cy- is each independently substituted by 1 to 3 R cx replace; R L21 、R L22 、R cx are each independently selected from hydrogen, halogen, -OR L2a 、-N(R L2a )2、-C(O)R L2a 、-S(O)2R L2a 、-C(O)N(R L2a )2、-SO2N(R L2a )2、-N(R L2a )SO2R L2b 、-N(R L2a )COR L2b 、-(CH2) y -CO-(N(Me)CH2C(O)) m -OR L2a 、-(CH2) y -CO-(N(Me)CH2C(O)) m -NHR L2a 、-(CH2) y -CONH-(CH2CH2O) m -R L2a 、-(CH2) y -NHCO-(CH2CH2O) m -R L2a 、-(CH2) y -NHCOCH2(OCH2CH2)OR L2a 、-(CH2) y -NH(COCH2(N(Me)) m -R L2a 、-(CH2) y -NHCO-(CH2CH2O) m -R L2a 、-(CH2CH2O) m -R L2a 、-(COCH2N(Me)) m -R L2a 、-COCH2(OCH2CH2) m -OR L2a 、-CO-(CH2CH2O) m -R L2a and R L2a Optionally substituted -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, 3-8 membered cycloalkyl, 4-10 membered heterocycloalkyl, 6-10 membered aryl or 3-10 membered heteroaryl; m is a natural integer from 0 to 8; y is selected from 0, 1, 2, 3 and 4; Each R L2a 、R L2b Each is independently selected from hydrogen, halogen, -CN, -OH, -NH2, -N(Me)2, -CO2H, -C(O)NH2 and -C 1-6 alkyl.

18. The ligand-drug conjugate according to any one of claims 2 to 14 and 16, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts or solvates, wherein L 2 -(CH2) n -; in, n is a natural integer from 0 to 50; for example, 0, 1, 2, 3, 4, 5, 6, 7 or 8; L 2 Any methylene unit in each of the following structural units is optionally replaced by a 4- to 6-membered heterocyclyl group, a 3- to 6-membered cycloalkylene group, -C(O)-, -NR L22 -、-O-、 R L22 are each independently selected from hydrogen, -OR L2a 、-C(O)R L2a 、-S(O)2R L2a 、-C(O)N(R L2a )2、-SO2N(R L2a )2、-(CH2) y -CO-(N(Me)CH2C(O)) m -OR L2a 、-(CH2) y -CO-(N(Me)CH2C(O)) m -NHR L2a 、-(CH2) y -CONH-(CH2CH2O) m -R L2a 、-(CH2) y -NHCOCH2(OCH2CH2)OR L2a 、-(CH2CH2O) m -R L2a 、-(COCH2N(Me)) m -R L2a 、-COCH2(OCH2CH2) m -OR L2a 、-CO-(CH2CH2O) m -R L2a and R L2a Optionally substituted -C 1-6 alkyl; m is a natural integer from 0 to 8; y is 0, 1, 2, 3, or 4; R L2a Each is independently selected from hydrogen, halogen, -CN, -OH, -NH2, -N(Me)2, -CO2H, -C(O)NH2 and -C 1-6 alkyl.

19. The ligand-drug conjugate according to any one of claims 2 to 14 and 16, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein L 2 Selected from: in, n1, n2, n3, n4, m are each independently selected from a natural number from 0 to 8; for example, 0, 1, 2, 3, 4, 5, 6, 7 or 8; n5 and n6 are each independently selected from 0 or 1; -Cy- is a 4- to 6-membered heterocyclylene or a 3- to 6-membered cycloalkylene; preferably, -Cy- is More preferably, -Cy- is More preferably, for More preferably, for Among them, c and L 1a Connected, d and L 3 connected.

20. The ligand-drug conjugate according to any one of claims 2 to 14, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts or solvates, wherein Selected from: n1, n2, n3, n4, and m are each independently selected from natural numbers from 0 to 8; n5 and n6 are each independently selected from 0 or 1; -Cy- is a 4- to 6-membered heterocyclylene group or a 3- to 6-membered cycloalkylene group; preferably, -Cy- is selected from: More preferably, -Cy- is selected from 21. The ligand-drug conjugate according to any one of claims 2 to 14, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts or solvates, wherein Selected from: More preferably, Selected from:

22. The ligand-drug conjugate according to any one of claims 2 to 14 and 16 to 21, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein L 3 Selected from Val, D-Val, Phe, Lys, Leu, Ile, Gly, Ala, D-Ala, Cit, Asp, Asn, Glu, Gln, Val-Cit, Val-Ala, Val- Lys, Val-Lys(Ac), Phe-Lys, Phe-Lys(Ac), Leu-Lys, Leu-Lys(Ac), Ala-Ala, Ala-Lys, D-Ala-Ala, Gly-Glu, Glu-Gly, Gly-Asp, Asp-Gly, Gly-Asn, Val-Glu, Val-Asp, Asn-Asn, Asn-Asn-Gly, Asp-Gl u, Gly-Gly-Glu, Glu-Gly-Gly, Gly-Glu-Gly, Gly-Gly-Asp, Gly-Gly-Asn, Gly-Ala-Ala, Gly-Val-A la, Gly-Val-Cit, Glu-Val-Cit, Ala-Ala-Ala, Ala-(D-Ala)-Ala, Ala-Ala-Asn, Ala-(D-Ala)-Asn ,Ala-Ala-Asp,Val-Lys-Gly,D-Val-Leu-Lys,Gly-Gly-Arg,Gly-Gly-Gly,Lys-Ala-Asn,Lys-Ala- Ala, Gly-Phe-Gly, Gly-Gly-Phe, Asn-Pro-Val, Ala-Lys-Gly, Gly-Lys-Gly, (Gly)3-Gly, (Gly)2- Phe-Gly, (Gly)2-Glu-Gly, Lys-(Ala)2-Asn, Lys-(Ala)2-Asp, (Ala)2-Pro-Val, (Ala)2-Pro-Nva, or any combination of the above fragments.

23. The ligand-drug conjugate according to any one of claims 2 to 14 and 16 to 21, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein L 3 Selected from Lys, Gly, Asp, Asn, Glu, Gln, Val-Cit, Val-Ala, Ala-Ala, Gly-Glu, Glu-Gly, Gly-Asp, Gly-Asn, Asn-Asn, As n-Asn-Gly, Asp-Glu, Asp-Gly, Gly-Glu-Gly, Glu-Gly-Gly, Lys-Ala-Ala, (Gly)2-Phe-Gly, (Gly)2-Glu-Gly, Or any combination of the above fragments; preferably, L 3 Selected from Lys, Gly, Val-Ala, Ala-Ala, Gly-Glu, Glu-Gly, Gly-Asp, Gly-Asn, Asn-Asn, Asp-Glu, Gly-Gly-Phe-Gly, Or any combination of the above fragments; More preferably, L 3 Selected from Val-Cit, Ala-Ala, Gly-Glu, Glu-Gly, Asn-Asn, Asn-Asn-Gly, Asp-Glu, Gly-Glu-Gly, Glu-Gly-Gly, Lys-Ala-Ala, (Gly)2-Glu-Gly, (Gly)2-Phe-Gly More preferably, L 3 Selected from Ala-Ala, Glu-Gly.

24. The ligand-drug conjugate according to any one of claims 2 to 14, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, in, Selected from: Preferably, Selected from More preferably, Selected from 25. The ligand-drug conjugate according to any one of claims 2 to 14 and 16 to 24, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: Tr does not exist or is in R Tr 、R Tr1 and R Tr2 independently selected from hydrogen, halogen, -NO2, -CN, -OH, -NH2, -CO2H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)NH2, -CH2CO-(N(Me)CH2C(O)) z -NHMe, -(CH2CH2O) z -H, -CONH-(CH2CH2O) z -H; z is a natural number from 0 to 8; for example, 0, 1, 2, 3, 4, 5, 6, 7, or 8.

26. The ligand-drug conjugate according to any one of claims 2 to 14 and 16 to 24, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein Tr is absent or is 27. The ligand-drug conjugate according to any one of claims 2 to 14 and 16 to 26, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein the structure represented by formula (IIa) or formula (IIb) is selected from:

28. The ligand-drug conjugate according to any one of claims 3 to 27, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein the ligand-drug conjugate is selected from: wherein Ab and q are as defined in claim 3.

29. The ligand-drug conjugate according to any one of claims 3 to 28, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein Ab is a target-binding polypeptide, antibody, or antigen-binding fragment thereof, in particular an antibody or antigen-binding fragment thereof.

30. The ligand-drug conjugate according to any one of claims 3 to 28, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: In Ab, the antibody is selected from one or more of the following: (1) Fully human antibodies, humanized antibodies, mouse antibodies and chimeric antibodies; (2) front resistance; (3) Bispecific antibodies and multispecific antibodies; (4) monoclonal antibodies and polyclonal antibodies; and (5) IgG antibodies.

31. The ligand-drug conjugate according to any one of claims 3 to 28, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: In Ab, the antigen-binding fragment is selected from the group consisting of: Fab, Fab', F(ab')2, Fv, scFv, Fd, dAb, VHH and complementarity determining region (CDR) fragments.

32. The ligand-drug conjugate according to any one of claims 3 to 28, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein Ab is a monoclonal antibody.

33. The ligand-drug conjugate according to any one of claims 3 to 28, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein Ab is an antibody or antigen-binding fragment thereof targeting HER2, HER3, B7H3, TROP2, Claudin18.2, GPC-3, ADAM9, CD30, CD33, CD70, GPC3, CEACAM5, and EGFR; preferably an anti-HER3 antibody or antigen-binding fragment thereof, for example, Patritumab or a variant thereof; an anti-GPC-3 antibody or antigen-binding fragment thereof, for example, Codrituzumab or a variant thereof; an anti-CEACAM5 antibody or antigen-binding fragment thereof, for example, Tusamitamab or a variant thereof.

34. The ligand-drug conjugate according to any one of claims 3 to 33, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: Ab contains: (I) HCDR1, HCDR2 and HCDR3 with amino acid sequences as shown in SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, respectively, and LCDR1, LCDR2 and LCDR3 with amino acid sequences as shown in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, respectively; or (II) HCDR1, HCDR2, and HCDR3 with amino acid sequences as shown in SEQ ID NO: 11, SEQ ID NO: 12, and SEQ ID NO: 13, respectively, and LCDR1, LCDR2, and LCDR3 with amino acid sequences as shown in SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16, respectively; or (III) the HCDR1, HCDR2 and HCDR3 of the amino acid sequences are shown in SEQ ID NO: 21, SEQ ID NO: 22 and SEQ ID NO: 23, respectively, and the light chain variable region comprises the LCDR1, LCDR2 and LCDR3 of the amino acid sequences are shown in SEQ ID NO: 24, SEQ ID NO: 25 and SEQ ID NO: 26, respectively.

35. The ligand-drug conjugate according to any one of claims 3 to 34, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: Ab contains: (I) a heavy chain variable region having an amino acid sequence as set forth in SEQ ID NO:7, or a sequence having at least 95%, 96%, 97%, 98% or 99% identity thereto, and a light chain variable region having an amino acid sequence as set forth in SEQ ID NO:8, or a sequence having at least 95%, 96%, 97%, 98% or 99% identity thereto; or (II) a heavy chain variable region having an amino acid sequence as set forth in SEQ ID NO: 17, or a sequence at least 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region having an amino acid sequence as set forth in SEQ ID NO: 18, or a sequence at least 95%, 96%, 97%, 98% or 99% identical thereto; or (III) a heavy chain variable region having an amino acid sequence as set forth in SEQ ID NO: 27, or a sequence having at least 95%, 96%, 97%, 98% or 99% identity thereto, and a light chain variable region having an amino acid sequence as set forth in SEQ ID NO: 28, or a sequence having at least 95%, 96%, 97%, 98% or 99% identity thereto.

36. The ligand-drug conjugate according to any one of claims 3 to 35, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: Ab contains: (I) a heavy chain variable region with an amino acid sequence as shown in SEQ ID NO: 7, and a light chain variable region with an amino acid sequence as shown in SEQ ID NO: 8; or (II) a heavy chain variable region with an amino acid sequence as shown in SEQ ID NO: 17, and a light chain variable region with an amino acid sequence as shown in SEQ ID NO: 18; or (III) a heavy chain variable region with an amino acid sequence as shown in SEQ ID NO: 27, and a light chain variable region with an amino acid sequence as shown in SEQ ID NO:

28.

37. The ligand-drug conjugate according to any one of claims 3 to 36, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: q is an integer or decimal from 2 to 8, for example, 2, 3, 4, 5, 6, 7, 8, 7.81 or 7.

62.

38. The ligand-drug conjugate according to any one of claims 3 to 37, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts or solvates, selected from wherein q is as defined in claim 3 or 37 above.

39. A compound represented by formula (IVa) or (IVb), or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof: in: R 1a , X, Y, Z, R 2a 、R 1b 、R 2b 、R 3b-1 and t as defined in claim 1 and any one of claims 4 to 14; L 2 、L 3 and Tr is as defined in claim 2 and any one of claims 17-19, 22-23; L 1 For the connection unit.

40. A compound according to formula (IVa) or (IVb) according to claim 39, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein L 1 Selected from:

41. A compound according to formula (IVa) or (IVb) according to claim 39, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein L 1 Selected from: L 1 Selected from: Preferably, L 1 Selected from 42. A compound of formula (IVa) or (IVb) according to claim 39, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: Selected from: n1, n2, n3, n4, and m are each independently selected from natural numbers from 0 to 8; n5 and n6 are each independently selected from 0 or 1; -Cy- is a 4- to 6-membered heterocyclylene or a 3- to 6-membered cycloalkylene; preferably, -Cy- is selected from: More preferably, -Cy- is selected from Preferably, Selected from: More preferably, Selected from:

43. A compound of formula (IVa) or (IVb) according to claim 39, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: Selected from: More preferably, Selected from:

44. A compound of formula (IVa) or (IVb) according to claim 39, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, selected from 45. A compound represented by formula (Va) or formula (Vb), or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof: in, R 3b Selected from -(C(R 3b-1 )2) t -H、-(C(R 3b-1 )2) t -OR 3b-2 、-(C(R 3b-1 )2) t -N(R 3b-2 )2、-(C(R 3b-1 )2) t -SR 3b-2 and -(C(R 3b-1 )2) t -S(O)2R 3b-2 ; Y, Z, R 1a 、R 2a 、R 1b 、R 2b 、R 3b-1 、R 3b-2 and t as defined in claim 1 and any one of claims 4 to 14.

46. The compound of formula (Va) or (Vb) according to claim 45, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: R 3b Selected from -(C(R 3b-1 )2) t -OR 3b-2 and -(C(R 3b-1 )2) t -N(R 3b-2 )2; preferably, R 3b Selected from -(C(R 3b-1 )2) t -OR 3b-2 ; Optionally, wherein R 1a selected from hydrogen; Y is selected from -C 1-6 alkylene; Z is absent or selected from -O-; R 2a Selected from hydrogen and -C 1-6 alkyl; R 1b 、R 2b are each independently selected from hydrogen, F, Cl and methyl; Or, R 1b With R 2b and the atoms to which it is attached together form a 5-membered heterocyclic ring, wherein the 5-membered heterocyclic ring is R 3b selected from -(C(R 3b-1 )2) t -OR 3b-2 ; t is selected from 2 and 3; R 3b-1 are each independently selected from hydrogen and methyl; R 3b-2 Selected from hydrogen.

47. A compound of formula (Va) or (Vb) according to claim 45, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: The compound is selected from:

48. A compound of formula (Va) or (Vb) according to claim 45, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: The compound is selected from:

49. A pharmaceutical composition comprising the ligand-drug conjugate of any one of claims 1-38, the compound of any one of claims 39-48, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

50. A pharmaceutical formulation comprising the ligand-drug conjugate of any one of claims 1 to 38, the compound of any one of claims 39 to 48, or a tautomer, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof.

51. Use of the ligand-drug conjugate according to any one of claims 1 to 38, the compound according to any one of claims 39 to 48, or its tautomers, enantiomers, diastereomers, or mixtures thereof, or its pharmaceutically acceptable salts or solvates, or the pharmaceutical composition according to claim 49, or the pharmaceutical preparation according to claim 50 in the preparation of a drug for preventing or treating a disease associated with abnormal cell activity.

52. The use according to claim 51, wherein the disease associated with abnormal cell activity is cancer.

53. The method of claim 52, wherein the cancer is a solid tumor or a non-solid tumor, for example, selected from esophageal cancer (e.g., esophageal adenocarcinoma and esophageal squamous cell carcinoma), brain tumor, lung cancer (e.g., small cell lung cancer and non-small cell lung cancer), squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, kidney cancer, non-Hodgkin's lymphoma, central nervous system tumors (e.g., glioma, glioblastoma multiforme, glioma or sarcoma), prostate cancer or thyroid cancer.

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