A steroid compound and its conjugate

Abstract

A steroidal compound and its conjugate, specifically the compound and its conjugate or its tautomer, mesomer, racemate, enantiomer, diastereoisomer, or mixture thereof, or its pharmaceutically acceptable salt; and the preparation method and application of the compound and its conjugate.

Description

Technical Field

[0001] The present application relates to the field of biomedicine, and specifically to a steroid compound and a conjugate thereof. Background Art

[0002] Inflammation is an adaptive response triggered by a variety of harmful stimuli and conditions, and it underlies many human immune system-related diseases. Steroidal compounds are a class of anti-inflammatory drugs that have the potential to influence immune system function and treat or prevent diseases and / or symptoms related to glucocorticoid receptor signaling. However, some existing steroidal compounds have weak anti-inflammatory effects, while others may have many undesirable side effects. Therefore, there is an urgent need to further develop antibody-drug conjugates and steroidal compounds based on multiple steroids as drugs with better efficacy or improved safety. Summary of the Invention

[0003] The present application provides a 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 the ability to affect the activity of immune cells; (2) having a targeting effect on its conjugate; (3) having plasma stability; (4) having biosafety; (5) having the ability to affect the release of cytokines from immune cells; (6) having the ability to affect the transcription of IFN signaling pathway response genes; (7) having the ability to affect the degree of skin fibrosis; (8) having the ability to affect the number and / or proportion of dendritic cells; (9) having the ability to affect the collagen content of the skin; (10) having the ability to affect the expression level of GRE; (11) having the ability to affect the release of monocyte cytokines; (12) having the ability to affect contact hypersensitivity; (13) having the ability to affect skin swelling and (14) having the ability to affect arthritis symptoms.

[0004] The application provides immunoconjugates containing glucocorticoid receptor agonists connected with proteins (e.g., antibodies or their Fabs and soluble receptor proteins). In certain embodiments, antibodies or their Fabs are people, humanized, chimeric or mouse. In certain embodiments, proteins (e.g., antibodies, their Fabs or soluble receptor proteins) can be combined with targets on cell surfaces and become internalized.

[0005] In one aspect, the present application provides a compound of formula I:

[0006]

[0007] or its tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein,

[0008] R1, R2, R3, R4 and R5 are each independently any group;

[0009] B does not exist or B is any group;

[0010] W does not exist or W is any group;

[0011] A1 is a substituted benzene ring;

[0012] Each CR4R5 unit is independently unsubstituted or substituted by a member selected from the group consisting of -O-, -S-, -NR-, -S(O)-, -S(O)2-, -C(=O)-, -C=C-, and -C≡C-, wherein R4 and R5 may together form an optionally substituted cycloalkyl, an optionally substituted heterocyclyl, an optionally substituted bridged ring group, an optionally substituted heterobridged ring group, an optionally substituted spirocyclyl, and an optionally substituted heterospirocyclyl, and n is any integer from 1 to 20;

[0013] X is selected from: -O-, -S-, -NR-;

[0014] Y1 is any group, m is any integer from 0 to 4;

[0015] wherein the substituents are selected from the group consisting of protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O)OR, -S(O)2NHR, -S(O) )2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy, wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0016] In another aspect, the present invention provides a compound represented by the following formula or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

[0017]

[0018]

[0019] In another aspect, the present application provides a compound of formula IIa or IIb:

[0020]

[0021] or its tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein,

[0022] R1, R2, R3, R4 and R5 are each independently any group;

[0023] B does not exist or B is any group;

[0024] W does not exist or W is any group;

[0025] A2 is a substituted benzene ring;

[0026] Each CR4R5 unit is independently unsubstituted or substituted by a member selected from the group consisting of -O-, -S-, -NR-, -S(O)-, -S(O)2-, -C(=O)-, -C=C-, and -C≡C-, wherein R4 and R5 may together form an optionally substituted cycloalkyl, an optionally substituted heterocyclyl, an optionally substituted bridged ring group, an optionally substituted heterobridged ring group, an optionally substituted spirocyclyl, and an optionally substituted heterospirocyclyl, and n is any integer from 1 to 20;

[0027] X is selected from: -O-, -S-, -NR-;

[0028] Y1 is any group, m is any integer from 0 to 4;

[0029] Y2 is selected from -O-, -S- and -NR-;

[0030] wherein the substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O)OR, -S(O)2NHR, -S( O)2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy; wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0031] On the other hand, the present application provides a conjugate comprising the aforementioned compound or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof.

[0032] On the other hand, the present application provides a pharmaceutical composition comprising the aforementioned compound or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof and / or the aforementioned conjugate, and optionally a pharmaceutically acceptable carrier.

[0033] On the other hand, the present application provides a method for influencing immune system function, comprising administering to a subject the aforementioned compound or its tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, the aforementioned conjugate and / or the aforementioned pharmaceutical composition.

[0034] Those skilled in the art can easily discern other aspects and advantages of the present application from the detailed description below. In the detailed description below, only exemplary embodiments of the present application are shown and described. As will be appreciated by those skilled in the art, the content of the present application enables those skilled in the art to modify the disclosed specific embodiments without departing from the spirit and scope of the invention to which the present application relates. Accordingly, the description in the specification of the present application is merely exemplary and not restrictive. DETAILED DESCRIPTION

[0035] The following describes the implementation of the present invention through specific embodiments. People familiar with this technology can easily understand other advantages and effects of the present invention from the contents disclosed in this specification.

[0036] Definition of terms

[0037] In this application, the term "glucocorticoid" generally refers to a naturally occurring steroid hormone or a synthetic steroid hormone that interacts with the glucocorticoid receptor.

[0038] In the present application, the term "halogen" generally refers to fluorine, chlorine, bromine, and iodine, for example, it can be fluorine or chlorine.

[0039] In the present application, the term "alkyl" generally refers to a residue derived from an alkane by removing a hydrogen atom. An alkyl group can be substituted or non-substituted, substituted or non-substituted. The term "alkyl" generally refers to a saturated straight or branched aliphatic hydrocarbon group having a residue derived from the same carbon atom or two different carbon atoms of the parent alkane by removing a hydrogen atom, and can be a straight or branched group containing 1 to 20 carbon atoms, for example, a chain alkyl group containing 1 to 12 carbon atoms, for example, 1 to 6 carbon atoms. Non-limiting examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, propyl, butyl, etc. The alkyl group may be substituted or unsubstituted, substituted or unsubstituted, for example, when substituted, the substituent may be substituted at any available point of attachment, and the substituent may be independently selected from one or more substituents selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo, for example, hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H or C 1-6 Aliphatic group.

[0040] In this application, the term "alkylene" generally refers to a saturated straight-chain or branched aliphatic hydrocarbon group having two residues derived from the same carbon atom or two different carbon atoms of an alkane group, and can be a straight-chain or branched group containing 1 to 20 carbon atoms. For example, the term "methylene" can refer to a residue derived from a group containing 1 carbon atom by removing two hydrogen atoms. The methylene group can be substituted or unsubstituted, substituted or unsubstituted; for example, an alkylene group containing 1 to 12 carbon atoms, for example, an alkylene group containing 1 to 6 carbon atoms. Non-limiting examples of alkylene groups include, but are not limited to, methylene (-CH2-), 1,1-ethylene (-CH(CH3)-), 1,2-ethylene (-CH2CH2)-, 1,1-propylene (-CH(CH2CH3)-), 1,2-propylene (-CH2CH(CH3)-), 1,3-propylene (-CH2CH2CH2-), 1,4-butylene (-CH2CH2CH2CH2-), and 1,5-butylene (-CH2CH2CH2CH2CH2-), and the like. Alkylene may be substituted or unsubstituted, substituted or unsubstituted, for example, when substituted, the substituent may be substituted at any available point of attachment, and the substituent may be independently selected from one or more substituents selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio and oxo, for example, hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H or C 1-6 Aliphatic groups. Methylene or alkylene groups may be substituted or unsubstituted.

[0041] In this application, the term "alkenyl" generally refers to a straight or branched hydrocarbon group containing one or more double bonds. Illustrative examples of alkenyl include allyl, homoallyl, vinyl, crotyl, butenyl, pentenyl, and hexenyl. Illustrative examples of C2-6 alkenyl groups with one or more double bonds include butadienyl, pentadienyl, hexadienyl, and hexatrienyl, as well as branched forms thereof. The position of the unsaturated bond (double bond) can be at any position of the carbon chain. Alkenyl groups can be substituted or unsubstituted.

[0042] In this application, the term "alkenylene" generally refers to a residue derived from an alkene carbon atom by removing two hydrogen atoms. For example, it can be allylene, vinylene, butenylene, pentenylene and hexenylene. Alkenylene can be substituted or unsubstituted.

[0043] In the present application, the term "alkynyl" generally refers to an unsaturated straight-chain or branched alkynyl group, such as ethynyl, 1-propynyl, propargyl, butynyl, etc. Alkynyl groups can be substituted or unsubstituted.

[0044] In this application, the term "alkynylene" generally refers to a residue derived from a carbon atom of an alkyne by removing two hydrogen atoms. For example, it may be ethynylene, propynylene, propargylene, butynylene, etc. Alkynylenes may be substituted or unsubstituted.

[0045] In this application, the term "aryl" generally refers to a residue derived from an aromatic ring by removing a hydrogen atom. The term "aromatic ring" can refer to a 6- to 14-membered all-carbon monocyclic ring or a fused polycyclic ring (i.e., a ring sharing adjacent pairs of carbon atoms) with a conjugated π electron system, which can be 6- to 10-membered, such as benzene and naphthalene. The aromatic ring can be fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, wherein the ring connected to the parent structure is the aryl ring. Aryl groups can be substituted or unsubstituted. When substituted, the substituents can be one or more of the following groups independently selected from the following groups: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio. Aryl groups can be substituted or unsubstituted.

[0046] In this application, the term "arylene" generally refers to a residue derived from a carbon atom in an aromatic ring by removing two hydrogen atoms. Examples include phenylene and naphthylene. Arylene can be substituted or unsubstituted.

[0047] As used herein, the term "heteroaryl" generally refers to a residue derived from a carbon atom in a heteroaromatic ring by removing a hydrogen atom. The term "heteroaromatic ring" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms may be selected from the group consisting of oxygen, sulfur, and nitrogen. Heteroaryl groups may be 5 to 10-membered, and may be 5- or 6-membered, such as furanyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, and the like. The heteroaryl ring may be fused to an aryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the heteroaryl ring. Heteroaryl can be optionally substituted or unsubstituted. When substituted, the substituent can be one or more of the following groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio. Heteroaryl can be substituted or unsubstituted.

[0048] In this application, the term "heteroarylene" generally refers to a residue derived from a carbon atom of a heteroaromatic ring by removing two hydrogen atoms. For example, it may be furanylene, thienylene, pyridylene, pyrrolylene, pyrimidylene, pyrazinylene, imidazolylene, tetrazolylene, etc. The heteroarylene group may be substituted or unsubstituted.

[0049] In this application, the terms "alicyclic" and "cycloalkyl" are used interchangeably. The term "alicyclic" generally refers to a residue derived by removing a hydrogen atom from the same carbon atom or multiple carbon atoms in an aliphatic ring. The term "cycloalkane" generally refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon, wherein the carbocyclic ring contains 3 to 20 carbon atoms, can contain 3 to 12 carbon atoms, can contain 3 to 10 carbon atoms, or can contain 3 to 8 carbon atoms. Non-limiting examples of alicyclic groups include cyclopropane, cyclobutane, cyclopentane, cyclopentenyl, cyclohexane, cyclohexenyl, cyclohexadienyl, cycloheptane, cycloheptatrienyl, cyclooctanyl, and the like; polycyclic carbocyclic rings can include spiro, fused, and bridged carbocyclic rings. Alicyclic groups can be substituted or unsubstituted. In this application, the term "carbocyclic" generally refers to a residue derived by removing a hydrogen atom from a carbon atom in the carbocyclic ring. The term "carbocycle" generally refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon ring, wherein the carbocycle contains 3 to 20 carbon atoms, may contain 3 to 12 carbon atoms, may contain 3 to 10 carbon atoms, and may contain 3 to 8 carbon atoms. Non-limiting examples of monocyclic carbocycles include cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexene, cyclohexadiene, cycloheptane, cycloheptatriene, cyclooctane, and the like; polycyclic carbocycles may include spirocyclic, fused, and bridged carbocycles. Carbocyclyl groups may be substituted or unsubstituted. In some cases, alicyclic and carbocyclic rings may be used interchangeably.

[0050] As used herein, the term "partially unsaturated" generally refers to a cyclic structure containing at least one double or triple bond between the ring molecules. The term "partially unsaturated" encompasses cyclic structures with multiple unsaturations, but is not intended to include aromatic or heteroaromatic rings as defined herein. The term "unsaturated" refers to a moiety having one or more degrees of unsaturation.

[0051] In this application, the terms "alicyclic group" and "cycloalkylene" are used interchangeably. The term "alicyclic group" generally refers to a residue derived from a carbon atom of an alicyclic ring by removing two hydrogen atoms. For example, it can be cyclopropylene, cyclobutane, cyclopentane, cyclopentenane, cyclohexane, cyclohexenane, cyclohexadienane, cycloheptanylene, cycloheptatrienylene, cyclooctanylene, etc.; polycyclic carbocycles can include spiro, fused, and bridged carbocycles. Alicyclic groups can be substituted or unsubstituted.

[0052] In this application, the terms "aliphatic heterocyclic group" and "heterocyclic group" can be used interchangeably. The term "aliphatic heterocyclic group" generally refers to a stable non-aromatic 3-membered to 7-membered monocyclic carbocyclic structure, a fused 7-membered to 10-membered bicyclic heterocyclic structure, or a bridged 6-membered to 10-membered bicyclic heterocyclic structure. These ring structures can be saturated or partially saturated. In addition to carbon atoms, these ring structures also contain one or more heteroatoms, wherein the heteroatoms can be selected from the following group: oxygen, sulfur, and nitrogen. For example, it contains 1-4 heteroatoms as defined above. When used to represent atoms in an aliphatic heterocyclic ring structure, the term "nitrogen" can include nitrogen that has undergone a substitution reaction. For example, a heterocyclic group may include a "heterocycloalkyl" group, which may refer to a stable non-aromatic 3- to 7-membered monocyclic alkane structure, a fused 7- to 10-membered bicyclic heterocyclic structure, or a bridged 6- to 10-membered bicyclic heterocyclic structure. In addition to carbon atoms, these cyclic structures may contain one or more heteroatoms, wherein the heteroatoms may be selected from the group consisting of oxygen, sulfur, and nitrogen. For example, the heterocycloalkyl group may contain 1 to 4 heteroatoms as defined above. The heterocycloalkyl group may be substituted or unsubstituted. The heterocyclic group may be substituted or unsubstituted.

[0053] In this application, the term "aliphatic heterocyclyl" and "heterocyclylene" can be used interchangeably. The term "aliphatic heterocyclyl" generally refers to a residue derived from a carbon atom of an aliphatic heterocycle by removing two hydrogen atoms. The aliphatic heterocyclyl group can be substituted or unsubstituted.

[0054] In this application, when describing cyclic structures such as cycloalkyl, cycloalkylene, heterocyclyl, heterocyclylene, aryl, arylene, heteroaryl, and heteroarylene, the prefix (e.g., C3-C 20 , C3-C7, C5-C6, etc.) generally refers to the number of ring atoms, or the range of the number of ring atoms, whether carbon atoms or heteroatoms. For example, as used in this application, the term "C5-C6 heterocyclyl" refers to a heterocyclyl group having 5 or 6 ring atoms. For another example, the term "C5-C6 heterocyclyl" as used in this application refers to a heterocyclyl group having 5 or 6 ring atoms. 10 "Heteroaryl" relates to heteroaryl groups having 5 to 10 ring atoms.

[0055] In this application, the term "ring-forming atoms" or "ring atoms" generally refers to atoms contained in a ring structure. For example, ring-forming atoms can be carbon atoms in a benzene ring or nitrogen atoms in a pyridine ring. When a ring-forming atom is attached to a hydrogen atom, the ring-forming atom can be substituted or unsubstituted.

[0056] As used herein, the term "each independently" generally refers to a variable being applicable in any instance, regardless of the presence or absence of variables with the same or different definitions in the same compound. For example, the variable may refer to the type or number of substituents in a compound, or the type of atoms in the compound. For example, if R occurs twice in a compound and R is defined as "independently carbon or nitrogen," both R may be carbon, both R may be nitrogen, or one R may be carbon and the other R may be nitrogen.

[0057] 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.

[0058] In this application, the term "substituted" generally refers to one or more hydrogen atoms in a group, for example up to 5, for example 1 to 3 hydrogen atoms, which 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.

[0059] When substituted, the substituents of an "optionally substituted" group may include, but are not limited to, one or more substituents independently selected from the following groups or groups specifically specified, alone or in combination: lower alkyl, lower alkenyl, lower alkynyl, lower alkanoyl, lower heteroalkyl, lower heterocycloalkyl, lower haloalkyl, lower haloalkenyl, lower haloalkynyl, lower perhaloalkyl, lower perhaloalkoxy, lower cycloalkyl, phenyl, aryl, aryloxy, lower alkoxy, lower haloalkoxy, oxo, lower acyloxy, carbonyl, carboxyl, thiazolin ... The substituents may be substituted or replaced with a substituted or replaced group. The substituents may be substituted or replaced with a substituted or replaced group. The substituents may be substituted or replaced with a substituted or replaced group. The substituents may be substituted or replaced with a substituted or replaced group. The substituents may be substituted or replaced with a substituted or replaced group. The substituents may be substituted or replaced with a substituted or replaced group. The substituents may be substituted or replaced with a substituted or replaced group. The substituents may be substituted or replaced with a substituted or replaced group. The substituents may be substituted or replaced with a substituted or replaced group. Optionally substituted groups can be incompletely substituted (e.g., -CH2CH3), completely substituted (e.g., -CF2CF3), monosubstituted (-CH2CH2F), or substituted at any level between complete and monosubstituted (e.g., -CH2CF3). When a substituent is listed without qualification as to substitution, both substituted and unsubstituted forms are included. When a substituent is identified as "substituted," the substituted form is specifically intended. Additionally, different groups of optional substituents for a particular moiety can be defined as needed; in these cases, the optional substitution is defined, usually immediately following the phrase "optionally substituted with..." The term "lower," when used in conjunction with an organic group or compound, means that the compound or group may be branched or unbranched, having no more than and including 7 carbon atoms, e.g., 1-4 carbon atoms. Lower alkyl represents, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl and branched pentyls, n-hexyl and branched hexyls.

[0060] In this application, the term 0 or more (e.g., 0 or more, 0 or 1, 0) methylene units are "replaced" generally means that when the structure contains 1 or more methylene units, the one or more methylene units may be unsubstituted or replaced by one or more groups that are not methylene (e.g., -NHC(O)-, -C(O)NH-, -C(O)-, -OC(O)-, -C(O)O-, -NH-, -O-, -S-, -SO-, -SO2-, -PH-, -P(=O)H-, -NHSO2-, -SONH-, -C(=S)-, -C(=NH)-, -N=N-, -C=N-, -N=C- or -C(=N2)-).

[0061] In this application, as known to those skilled in the art, terms such as "alkyl", "alkenyl", "cycloalkyl", etc. may be preceded by an identifier to indicate the number of atoms present in the group in a particular case, for example, C1-C4 alkyl, C3-C7 cycloalkyloxy, C1-C4 alkylcarbonylamino, etc., and the subscript number after "C" indicates the number of carbon atoms present in the group. For example, C3 alkyl refers to an alkyl group with three carbon atoms (e.g., n-propyl, isopropyl); C 1-10 In the examples, the members of the group may have any number of carbon atoms falling within the range of 1-10.

[0062] One or more hydrogen atoms, for example up to 5, for example 1 to 3, in a group are independently replaced by a corresponding number of substituents. The substituents are only in their possible chemical positions, and a person skilled in the art can determine (by experiment or theory) which substitutions are possible or impossible 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.

[0063] In this application, the term "compound" generally refers to a substance with 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 less than 500, a compound with a molecular weight of less than 1000, a compound with a molecular weight of more than 1000, or a compound with a molecular weight of more than 10,000 or more than 100,000. In this application, a compound can also refer to a compound connected by chemical bonds, for example, it can be a compound in which one or more molecules with a molecular weight of less than 1000 are connected to a biomacromolecule through a chemical bond, 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 less than 1000, a compound in which a protein is connected to one or more molecules with a molecular weight of less than 10,000, or a compound in which a protein is connected to one or more molecules with a molecular weight of less than 100,000.

[0064] The pharmaceutical composition may be in the form of a sterile injectable aqueous or oily suspension for intramuscular and subcutaneous administration. The suspension may be formulated using suitable dispersants or wetting agents and suspending agents as described above according to known techniques. Sterile injectable formulations may also be sterile injectable solutions or suspensions prepared in a nontoxic, parenterally acceptable diluent or solvent, such as a solution prepared in 1,3-butanediol. Furthermore, sterile fixed oils may conveniently be used as a solvent or suspending medium. For example, any blended fixed oil, including synthetic mono- or diglycerides, may be used. Furthermore, fatty acids, such as oleic acid, may also be used to prepare injectable formulations.

[0065] In the present application, the compound of the present application includes its tautomer, mesomorph, racemate, enantiomer and / or diastereomer of compound.In the present application, the term "diastereomer" generally refers to a stereoisomer with two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers can have different physical properties, for example, melting point, boiling point, spectral properties and reactivity. In the present application, the term "tautomer" or "tautomeric form" is used interchangeably, generally referring to structural isomers of different energies that can be converted to each other by low energy barriers (low energy barrier). For example, proton tautomers (protontautomers) (also referred to as prototropic tautomers (prototropic tautomers)) include the mutual conversions carried out by proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers (valence tautomers) include the mutual conversions carried out by the reorganization of some bonding electrons. As used herein, the term "meso" generally refers to a molecule containing asymmetric atoms but possessing symmetry such that the total optical rotation is zero. The term "racemate" or "racemic mixture" refers to a composition composed of equimolar amounts of two enantiomeric species.

[0066] In the present application, some atoms of the compounds of the present application may appear in more than one isotopic form. For example, hydrogen may appear in the form of protium ( 1 H), deuterium ( 2 H) and tritium ( 3 H), carbon can exist in three different isotopes ( 12 C. 13 C and 14 C) naturally occurring. Examples of isotopes that can be incorporated into the compounds of the present application also include, but are not limited to 15 N. 18 O. 17 O. 18 F. 32 P. 33 P. 129 I. 131 I. 123 I. 124 I. 125 I, or similar isotopes. Therefore, the compounds of the present application may be enriched in one or more of these isotopes relative to the natural abundance of these isotopes. As known to those skilled in the art, such isotopically enriched compounds can be used for a variety of purposes. For example, using heavy isotopes such as deuterium ( 2 Deuterium (H) substitution may offer certain therapeutic advantages, possibly due to greater metabolic stability. 2The natural abundance of deuterium (H) is about 0.015%. Therefore, there is one deuterium atom for approximately every 6,500 hydrogen atoms in nature. Therefore, the deuterium-containing compounds of the present invention have a deuterium abundance greater than 0.015% at one or more positions (as the case may be). 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 whose remaining portions are consistent with the present structure, except that hydrogen atoms are replaced by deuterium or tritium, or carbon atoms are replaced by carbon 13 or carbon 14, are within the scope of the present application.

[0067] In this application, "isomers" generally refer to different compounds with the same molecular formula. "Stereoisomers" generally refer to isomers that differ only in the arrangement of their atoms in space. In this application, the term "isomer" includes any and all geometric isomers and stereoisomers. For example, "isomers" include geometric double bond cis and trans isomers, also known as E- and Z-isomers; R- and S-enantiomers; diastereomers, (d)-isomers and (l)-isomers, racemic mixtures thereof; and other mixtures thereof falling within the scope of this disclosure.

[0068] In this application, "enantiomer" generally refers to a pair of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. Where appropriate, the term "(±)" is used to represent a racemic mixture. "Diastereomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. Absolute stereochemistry is defined according to the Cahn-Ingold-Prelog RS system. When a compound is a pure enantiomer, the stereochemistry at each chiral carbon can be specified by R or S. A split compound with unknown absolute configuration can be specified as (+) or (-) based on the direction (right-handed or left-handed) in which it rotates plane polarized light at the wavelength of the sodium D line. Certain compounds described herein contain one or more asymmetric centers and can therefore produce enantiomers, diastereomers, and other stereoisomeric forms that can be defined according to absolute stereochemistry, such as (R)- or (S)-. The chemical entities, pharmaceutical compositions and methods of the present application are intended to include all such possible isomers, including racemic mixtures, optically pure forms and intermediate mixtures. Optically active (R)- and (S)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. When the compounds described herein contain olefinic double bonds or other geometric asymmetric centers, and unless otherwise indicated, the compounds are intended to include both E and Z geometric isomers.

[0069] In this application, the term "enantiomeric purity" generally refers to the relative amount of a particular enantiomer present relative to the other enantiomer, expressed as a percentage. For example, if a compound that may have (R)- or (S)- isomeric configurations exists as a racemic mixture, then the enantiomeric purity with respect to either (R)- or (S)- isomer is about 50%. If one configuration of the compound is predominant over the other, for example, 80% (S)- and 20% (R)-, then the enantiomeric purity of the compound with respect to the (S)- isomer is 80%. The enantiomeric purity of a compound can be determined by a variety of methods known in the art, including but not limited to chromatography using a chiral support, polarization measurement of polarized light rotation, nuclear magnetic resonance spectroscopy using chiral shift reagents (including but not limited to chiral complexes containing lanthanides or Pirkle alcohols), or derivatization of the compound with a chiral compound such as Mosher's acid followed by chromatography or nuclear magnetic resonance spectroscopy.

[0070] In this application, the term "tautomer" generally refers to a type of isomer that includes two or more interconvertible compounds resulting from at least one formal migration of a hydrogen atom and at least one change in valence (e.g., a single bond to a double bond, a triple bond to a double bond, or a triple bond to a single bond, or vice versa). "Tautomerism" includes prototropic or proton-shift tautomerism, which is considered a subset of acid-base chemistry. "Prototropic tautomerism" or "proton-shift tautomerism" involves the migration of a proton with a change in bond order. The exact ratio of tautomers depends on a variety of factors, including temperature, solvent, and pH. When tautomerization is possible (e.g., in solution), a chemical equilibrium of tautomers can be reached. Tautomerism (i.e., a reaction that provides a pair of tautomers) can be catalyzed by an acid or base, or can occur without the action or presence of an external agent. Exemplary tautomerism includes, but is not limited to, keto-enol; amide-imide; lactam-lactim; enamine-imine; and enamine-(different)enamine tautomerism. A specific example of keto-enol tautomerism is the interconversion of pentane-2,4-dione and 4-hydroxypent-3-en-2-one tautomers. Another example of tautomerism is phenol-keto tautomerism. A specific example of phenol-keto tautomerism is the interconversion of pyridine-4-ol and pyridine-4(1H)-one tautomers.

[0071] As used herein, the term "pharmaceutical composition" generally refers to a mixture containing one or more compounds described herein, or their physiologically / pharmaceutically acceptable salts or prodrugs, and other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. Pharmaceutical compositions can facilitate administration to an organism, facilitating the absorption of the active ingredient and thereby exerting its biological activity. The preparation of conventional pharmaceutical compositions can be found in the Chinese Pharmacopoeia.

[0072] In this application, the term "pharmaceutically acceptable salt" or "pharmaceutically usable salt" generally refers to a salt of the compound or ligand-drug conjugate of the present application, or a salt of the compound described in the present application. Such salts may be safe and / or effective when used in mammals and may have the desired biological activity. The antibody-antibody drug conjugate compound of the present application may form a salt with an acid. Non-limiting examples of pharmaceutically acceptable salts include: hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, citrate, acetate, succinate, ascorbate, oxalate, nitrate, sorbate, hydrogen phosphate, dihydrogen phosphate, salicylate, hydrogen citrate, tartrate, maleate, fumarate, formate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, and p-toluenesulfonate.

[0073] In this application, the terms "immunoconjugate," "conjugate," "antibody-drug conjugate," or "ADC" generally refer to a compound or derivative thereof linked to a protein, such as a cell-binding agent (e.g., an anti-TNF-α antibody or fragment thereof), and defined by the following general formula: (SM-LQ)nA, wherein SM = a group derived from a small molecule glucocorticoid receptor agonist (e.g., a glucocorticoid), L = a linker, Q = a heterobifunctional group, a heterotrifunctional group, or absent, and A = a protein (e.g., an antibody or antigen-binding fragment thereof, an anti-TNF protein, an anti-TNF-α antibody or fragment thereof, a soluble receptor, or a soluble TNF receptor), and n = 1-10. An immunoconjugate can also be defined by the following general formula in reverse order: A-(QL-SM)n.

[0074] In the present application, the term "linker" generally refers to any chemical moiety that can be connected to a glucocorticoid by a protein (e.g., an antibody, an antibody fragment (e.g., Fab) or a functional equivalent). The linker may be sensitive to cutting ("cleavable linker"), thereby promoting the release of the glucocorticoid. For example, under conditions where glucocorticoid and / or antibody remain active, such cleavable linkers may be sensitive to acid-induced cutting, light-induced cutting, peptidase-induced cutting, esterase-induced cutting and disulfide bond cutting. Alternatively, the linker may be substantially resistant to cutting ("non-cleavable linker").

[0075] In this application, the term "drug-to-antibody ratio" or "DAR" refers to the number of SMs (i.e., groups derived from a small molecule glucocorticoid receptor agonist (e.g., a glucocorticoid)) attached to A (i.e., a protein, such as an antibody or antigen-binding fragment thereof, an anti-TNF protein, an anti-TNF-α antibody or fragment thereof, a soluble receptor, or a soluble TNF receptor). Thus, in an immunoconjugate having the general formula (SM-LQ)nA, the DAR is defined by the variable "n."

[0076] When referring to a compound of formula (SM-LQ)nA representing an individual immunoconjugate, DAR generally refers to the number of SMs attached to the individual A (eg, n is an integer from 1 to 10).

[0077] When referring to a compound of formula (SM-LQ)nA representing a plurality of immunoconjugates, DAR generally refers to the average number of SMs attached to A (e.g., n is an integer or fraction from 1 to 10). Thus, by way of example, a compound of formula (SM-LQ)nA comprising a first immunoconjugate containing 3 SMs per A and a second immunoconjugate containing 4 SMs per A has a DAR (i.e., "n") of 3.5.

[0078] In the present application, non-cleavable linker is normally any chemical part that can connect glucocorticoid to antibody in a stable covalent manner, and can not come off under the classification listed for cleavable linker above.Therefore, non-cleavable linker is basically acid-resistant induced cutting, light-induced cutting, peptidase-induced cutting, esterase-induced cutting and disulfide bond cutting.In addition, non-cleavable refers to that under the condition that glucocorticoid and / or antibody do not lose its activity, the chemical bond in the joint or adjacent to the joint withstands the ability of the cutting induced by acid, light-labile cutting agent, peptidase, esterase or the chemistry of cutting disulfide bond or physiological compound.

[0079] Some cleavable linkers are cleaved by peptidases ("peptidase-cleavable linkers"). Only certain peptides are susceptible to cleavage inside or outside cells, see, for example, Trout et al., Proc. Natl. Acad. Sci. USA, 626-629 (1982) and Umemoto et al., Int. J. Cancer, 677-684 (1989). In addition, peptides are composed of α-amino acid units and peptide bonds, which are chemically amide bonds between the carboxylic acid of one amino acid and the amino group of a second amino acid. Other amide bonds (such as the bond between the carboxylic acid of lysine and the α-amino acid group) are understood not to be peptide bonds and are considered non-cleavable.

[0080] Some linkers are cleaved by esterases ("esterase-cleavable linkers"). Only certain esters can be cleaved by esterases present inside or outside the cell. Esters are formed by the condensation of carboxylic acids and alcohols. Simple esters are esters produced with simple alcohols (such as aliphatic alcohols) and small cyclic alcohols and small aromatic alcohols.

[0081] In some embodiments, the cleavable linker component may comprise a peptide containing 1 to 10 amino acid residues. In these embodiments, the peptide allows protease cleavage of the linker, thereby promoting the release of glucocorticoids when exposed to intracellular proteases (such as lysosomal enzymes) (Doronina et al. (2003) Nat. Biotechnol. [Nature Biotechnology] 21: 778-784). Exemplary peptides include, but are not limited to, dipeptides, tripeptides, tetrapeptides, and pentapeptides. Exemplary dipeptides include, but are not limited to, alanine-alanine (ala-ala); valine-citrulline (vc or val-cit); alanine-phenylalanine (af or ala-phe); phenylalanine-lysine (fk or phe-lys); phenylalanine-homolysine (phe-homolys); and N-methyl-valine-citrulline (Me-val-cit). Exemplary tripeptides include, but are not limited to, glycine-valine-citrulline (glv-val-cit) and glycine-glycine-glycine (gly-gly-gly).

[0082] The peptide can comprise naturally occurring amino acid residues and / or non-natural amino acid residues. The term "naturally occurring amino acid" generally refers to Ala, Asp, Cys, Glu, Phe, Gly, His, He, Lys, Leu, Met, Asn, Pro, Gin, Arg, Ser, Thr, Val, Trp and Tyr. By way of non-limiting example, "non-natural amino acids" (i.e., naturally occurring amino acids) include homoserine, homoarginine, citrulline, phenylglycine, taurine, iodotyrosine, selenocysteine, norleucine ("Nle"), norvaline ("Nva"), β-alanine, L-naphthylalanine or D-naphthylalanine, ornithine ("Orn"), etc. Peptides can be designed and optimized for enzymatic cleavage by specific enzymes (e.g., tumor-associated proteases, cathepsins B, C and D or plasmin proteases).

[0083] Amino acids can also include D- forms of natural amino acids and non-natural amino acids. "D-" refers to an amino acid with a "D" (dextrorotatory) configuration, as opposed to the configuration in naturally occurring ("L-") amino acids. Natural amino acids and non-natural amino acids can be commercially available (Sigma Chemical Co., Advanced Chemtech) or can be synthesized using methods known in the art.

[0084] In this application, the term "pharmaceutically acceptable carrier" generally refers to a carrier for administering therapeutic agents, such as antibodies or polypeptides, genes, and other therapeutic agents. The term refers to any pharmaceutical carrier that does not itself induce the production of antibodies harmful to the individual receiving the composition and that can be administered without excessive toxicity. Suitable carriers can be large, slowly metabolized macromolecules, such as proteins, polysaccharides, polylactic acids, polyglycolic acids, polyamino acids, amino acid copolymers, lipid aggregates, and inactivated viral particles. These carriers are well known to those skilled in the art. Pharmaceutically acceptable carriers in therapeutic compositions can include liquids such as water, saline, glycerol, and ethanol. Auxiliary substances, such as wetting agents or emulsifiers, pH buffering substances, etc., may also be present in these carriers.

[0085] In this application, the term "anti-TNFα protein" generally refers to a protein that is capable of (i) binding to TNFα and (ii) inhibiting the binding of soluble TNF-α to cell surface TNF receptors (p55 and / or p75) and / or lysing cells expressing surface TNFα or TNFα receptors in vitro in the presence of complement. Anti-TNFα proteins include, for example, anti-TNF antibodies or antigen-binding fragments thereof (e.g., adalimumab or infliximab) and soluble TNF receptors (e.g., etanercept).

[0086] In this application, the term "antibody" generally refers to an immunoglobulin that is reactive to a specified protein or peptide or a fragment thereof. The antibody can be an antibody from any class, including but not limited to IgG, IgA, IgM, IgD and IgE, and an antibody from any subclass (e.g., IgG1, IgG2, IgG3, and IgG4). The antibody can have a heavy chain constant region selected from, for example, IgG1, IgG2, IgG3, or IgG4. The antibody can also have a light chain selected from, for example, kappa (κ) or lambda (λ). The antibody of the present application can be derived from any species. The term "antibody" can include complete polyclonal antibodies, complete monoclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, fusion proteins comprising antibodies, and any other modified immunoglobulin molecules, as long as these antibodies exhibit the desired biological activity.

[0087] In this application, the term "antigen-binding fragment" generally refers to a portion of an antibody molecule that contains amino acids responsible for the specific binding between the antibody and the antigen. The portion of the antigen that is specifically recognized and bound by the antibody is referred to as an "epitope" as described above. As described above, an antigen-binding domain typically comprises an antibody light chain variable region (VL) and an antibody heavy chain variable region (VH); however, it does not necessarily comprise both. An Fd fragment, for example, has two VH regions and generally retains some antigen-binding functions of a complete antigen-binding domain. Examples of antigen-binding fragments of antibodies include (1) a Fab fragment, a monovalent fragment having a VL, VH, constant light chain (CL) and CH1 domains; (2) a F(ab′)2 fragment, a bivalent fragment having two Fab fragments connected by a disulfide bridge at the hinge region; (3) an Fd fragment having two VH and CH1 domains; (4) an Fv fragment having the VL and VH domains of a single arm of an antibody, (5) a dAb fragment (Ward et al., “Binding Activities of a Repertoire of Single Immunoglobulin Variable Domains Secreted From Escherichia coli,” Nature 341:544-546 (1989), which is incorporated herein by reference in its entirety), which has a VH domain; (6) isolated complementarity determining regions (CDRs); and (7) single-chain Fv (scFv), for example, derived from an scFV library.Although the two domains VL and VH of the Fv fragment are encoded by independent genes, they can be joined using recombinant methods by a synthetic linker, which allows them to be prepared as a single protein chain (called single-chain Fv (scFv)) in which the VL and VH regions pair to form a monovalent molecule (see, for example, Huston et al., "Protein Engineering of Antibody Binding Sites: Recovery of Specific Activity in an Anti-Digoxin Single-Chain Fv Analogue Produced in Escherichia coli," Proc. Natl. Acad. Sci. USA 85: 5879-5883 (1988)); (8) "VHH" refers to the variable antigen-binding domain of heavy chain antibodies from the Camelidae (camel, dromedary, llama, alpaca, etc.) (see Nguyen VK et al., 2000, The EMBO Journal, 19, 921-930; Muyldermans S., 2001, J. Biotechnol., 74, 277-302 and reviewed in Vanlandschoot P. et al., 2011, Antiviral Research 92, 389-407). VHHs are also referred to as nanobodies (Nb).

[0088] In this application, the term "variable region" or "variable domain" generally refers to the domain of the antibody heavy chain or light chain involved in the binding of the antibody to the antigen. In this application, the term "variable" generally refers to that some parts of the sequence of the variable domain of the antibody vary strongly, forming the binding and specificity of various specific antibodies to their specific antigens. Variability is not evenly distributed in the entire variable region of the antibody. It is concentrated in three segments in the light chain variable region and the heavy chain variable region, known as the complementary determining region (CDR) or hypervariable region (HVR), namely LCDR1, LCDR2, LCDR3, HCDR1, HCDR2 and HCDR3. The more highly conserved parts in the variable domain are called framework regions (FR). The variable domains of natural heavy and light chains each include four FR regions (H-FR1, H-FR2, H-FR3, H-FR4, L-FR1, L-FR2, L-FR3, L-FR4), most of which adopt a β-pleated sheet configuration and are connected by three CDR structure loop regions. The CDRs in each chain are held together by the FR regions and, together with the CDRs from the other chain, form the antibody antigen-binding site.

[0089] In the art, a variety of methods can be used to encode the variable region of an antibody or divide the CDR of an antibody, such as the Kabat numbering scheme and definition rules based on sequence variability (see, Kabat et al., Protein Sequences in Immunology, 5th ed., National Institutes of Health, Bethesda, MD (1991)), the Chothia numbering scheme and definition rules based on the position of the structural loop region (see, Al-Lazikani et al., J Mol Biol 273:927-48, 1997), the IMGT numbering scheme and definition rules based on the amino acid sequence alignment of germline V genes by efranc et al., Honneger's numbering scheme (AHo's), Martin numbering scheme, Gelfand numbering scheme, etc., see Mathieu Dondelinger et al., Understanding the Significance and Implications of Antibody Numbering and Antigen-Binding Surface / Residue Definition, Front. Immunol., 16 October 2018.

[0090] In this application, the term "percentage (%) sequence identity" generally refers to the matching ("hits") number of the amino acids that are consistent between two or more compared amino acid sequences compared to the number of amino acid residues that make up the total length of these amino acid sequences. In other words, using comparison, for two or more sequences, when these sequences are compared and compared for maximum correspondence (as measured using a sequence comparison algorithm known in the art), or when manually aligned and visually inspected, the percentage of identical amino acid residues (e.g., 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity) can be determined. Therefore, sequences that are compared to determine sequence identity can be distinguished by one or more amino acid substitutions, additions or deletions. Suitable programs for aligning protein sequences are well known to those skilled in the art. The percent sequence identity of a protein sequence can be determined, for example, using programs such as CLUSTALW, Clustal Omega, FASTA, or BLAST, for example using the NCBI BLAST algorithm (Altschul SF et al. (1997), Nucleic Acids Res. 25:3389-3402).

[0091] In this application, the term "antibody analog" is generally used in the broadest sense and specifically covers molecules that specifically bind to a target molecule with a single specificity and are structurally different from a natural antibody. For example, in the context of describing an anti-PD-1 antibody or an anti-PD-L1 antibody, the term "antibody analog" refers to an antibody that comprises a segment that is substantially identical to a portion of the amino acid sequence and has at least one of the following properties: (1) specifically binds to PD-1 or PD-L1 under appropriate binding conditions, (2) the ability to inhibit at least one biological activity of PD-1 or PD-L1. Typically, an antibody analog comprises conservative amino acid substitutions (or insertions or deletions) relative to the natural sequence. Analogs are typically at least 20 or 25 amino acids long, at least 50, 60, 70, 80, 90, 100, 150 or 200 amino acids long or longer, and can typically be as long as the full-length heavy or light chain of an antibody. Some examples include antibody analogs having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 substitutions compared to the germline amino acid sequence.

[0092] In this application, the term "effective amount" or "therapeutically effective amount" generally refers to an amount of a compound or pharmaceutical composition described herein that is sufficient to achieve the intended application described below, including but not limited to disease treatment. The therapeutically effective amount may vary depending on: the intended application (in vivo or in vitro); or the subject and disease condition being treated, for example, the weight and age of the subject, the severity of the disease condition; the mode of administration, etc., which can be readily determined by a person of ordinary skill in the art. The term also applies to doses that will induce a specific response in the target cells, such as platelet adhesion and / or cell migration. The specific dose will vary depending on, for example, the specific compound selected, the dosing regimen followed, whether it is administered in combination with other agents, the time of administration, the tissue to which it is administered, and the physical delivery system used to transport it.

[0093] As used herein, the term "in vivo" generally refers to events that occur within a subject's body.

[0094] As used herein, the term "in vitro" generally refers to an event occurring outside a subject. For example, an in vitro assay includes any assay performed outside a subject. In vitro assays include cell-based assays, which utilize living or dead cells. In vitro assays also include cell-free assays, which do not utilize intact cells.

[0095] In this application, the terms "treatment" and "treating" generally refer to a method of obtaining a beneficial or desired result, including but not limited to a therapeutic benefit. Therapeutic benefit includes but is not limited to eradication, inhibition, reduction or improvement of the underlying disorder being treated. In addition, therapeutic benefit is achieved by eradication, inhibition, reduction or improvement of one or more physiological symptoms associated with the underlying disorder, such that improvement is observed in the patient, but the patient may still suffer from the underlying disorder.

[0096] As used herein, the terms "prevention" and "preventing" generally refer to an approach to obtaining a beneficial or desired result, including, but not limited to, a prophylactic benefit. For prophylactic benefit, a pharmaceutical composition can be administered to a patient at risk for developing a particular disease or to a patient reporting one or more physiological symptoms of a disease, even if the disease has not yet been diagnosed.

[0097] In this application, the term "subject" or "patient" generally refers to humans (i.e., males or females of any age group, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or elderly adults)) and / or other primates (e.g., cynomolgus monkeys, rhesus monkeys); mammals, including commercially relevant mammals such as cattle, pigs, horses, sheep, goats, cats, and / or dogs; and / or birds, including commercially relevant birds such as chickens, ducks, geese, quail, and / or turkeys.

[0098] In this application, the term "about" or "approximately" generally refers to an acceptable error for a particular value determined by one of ordinary skill in the art, which depends in part on the manner in which the value is measured or determined. In certain embodiments, the term "about" or "approximately" generally refers to 1, 2, 3, or 4 standard deviations. In certain embodiments, the term "about" or "approximately" generally refers to within 50%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.05% of a given value or range.

[0099] In this application, the term "comprise" and its variations, including "contain", "include" and other forms, generally refer to the inclusion of other components, elements, values, steps, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0100] Figure 1: Inhibitory effects of the drug conjugates of the present invention on R848-induced IFNα, TNFα, IL-6, and IL-8 production in human peripheral blood mononuclear cells. 1a: R848-induced IFNα release in PBMCs; 1b: R848-induced TNFα release in PBMCs; 1c: R848-induced IL6 release in PBMCs; 1d: R848-induced IL8 release in PBMCs.

[0101] Figure 2 : Determination of bioactivity in a fluorescein isothiocyanate (FITC)-induced delayed-type IV hypersensitivity mouse model.

[0102] Figure 3: Arthritis score in the DBA / 1 mouse arthritis model induced by type II bovine collagen mixed with adjuvant. 3a: Arthritis score; 3b: AUC of arthritis score. Detailed Description of the Invention

[0104]

[0105] or its tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein,

[0106] R1, R2, R3, R4 and R5 are each independently any group;

[0107] B does not exist or B is any group;

[0108] W does not exist or W is any group;

[0109] A1 is a substituted benzene ring;

[0110] Each CR4R5 unit is independently unsubstituted or substituted by a member selected from the group consisting of -O-, -S-, -NR-, -S(O)-, -S(O)2-, -C(=O)-, -C=C-, and -C≡C-, wherein R4 and R5 may together form an optionally substituted cycloalkyl, an optionally substituted heterocyclyl, an optionally substituted bridged ring group, an optionally substituted heterobridged ring group, an optionally substituted spirocyclyl, and an optionally substituted heterospirocyclyl, and n is any integer from 1 to 20;

[0111] X is selected from: -O-, -S-, -NR-;

[0112] Y1 is any group, m is any integer from 0 to 4;

[0113] wherein the substituents are selected from the group consisting of protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O)OR, -S(O)2NHR, -S(O) )2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy, wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0114] In certain embodiments, the compound of Formula I or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt is selected from:

[0115]

[0116] in,

[0117] R1, R2, R3, R4 and R5 are each independently any group;

[0118] B does not exist or B is any group;

[0119] W does not exist or W is any group;

[0120] A1 is a substituted benzene ring;

[0121] Each CR4R5 unit is independently unsubstituted or substituted by a member selected from the group consisting of -O-, -S-, -NR-, -S(O)-, -S(O)2-, -C(=O)-, -C=C-, and -C≡C-, wherein R4 and R5 may together form an optionally substituted cycloalkyl, an optionally substituted heterocyclyl, an optionally substituted bridged ring group, an optionally substituted heterobridged ring group, an optionally substituted spirocyclyl, and an optionally substituted heterospirocyclyl, and n is any integer from 1 to 20;

[0122] X is selected from: -O-, -S-, -NR-;

[0123] Y1 is any group, m is 0 or 1;

[0124] wherein the substituents are selected from the group consisting of protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O)OR, -S(O)2NHR, -S(O) )2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy, wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0125] In certain preferred embodiments of the present invention, certain groups in the compounds of formula (I), (Ia), (Ib), (Ic) or pharmaceutically acceptable salts thereof are defined as follows, and the unmentioned groups are the same as those described in any embodiment of the present application (referred to as "in certain embodiments").

[0126] In certain embodiments, X is -O-, -S-, or -NH-.

[0127] In certain embodiments, each of the CR4R5 units is independently unsubstituted or substituted with a member selected from the group consisting of -O-, -S-, and -C(=O)-.

[0128] In certain embodiments, wherein said R4 and R5 are taken together to form an optionally substituted cycloalkyl or an optionally substituted heterocyclyl.

[0129] In certain embodiments, wherein said R4 and R5 are each independently selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; Wherein, when R4 and R5 contain methylene units, the methylene units of R4 and R5 are each independently not substituted, or the methylene units of R4 and R5 are each independently substituted by a group selected from the following groups: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene.

[0130] In certain embodiments, wherein said R4 and R5 are each independently selected from the following group: hydrogen, protium, deuterium, tritium, halogen, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R4 and R5 contain methylene units, said methylene units of said R4 and R5 are Each of the methylene units of R4 and R5 is independently not substituted, or each of the methylene units of R4 and R5 is independently substituted by a group selected from the following groups: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene.

[0131] In certain embodiments, the compound of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein R4 and R5 are each independently selected from: H, F, Cl, -OH, -NH2, C1-C6 alkyl, or R4 and R5 together form a C3-C6 cycloalkyl or a 3-6 membered heterocyclyl; and n is selected from 1, 2 or 3.

[0132] In certain embodiments, the compound of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein the -(CR4R5) n - are each independently selected from: -CH2-, -CH2CH2-,

[0133] In certain embodiments, wherein said Y1 is absent or selected from: protium, deuterium, tritium, halogen, -OR, -SR, -NHR, -N(R)2, -PHR, -P(R)2, -P(=O)HR, -P(=O)R2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0134] In certain embodiments, wherein said Y1 is absent or selected from: protium, deuterium, tritium, halogen, -OR, -SR, -NHR, -N(R)2, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy; wherein each R is independently selected from hydrogen, protium, deuterium, tritium, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0135] In certain embodiments, wherein said Y1 is absent or selected from the group consisting of: hydroxyl, thiol, amino, and optionally substituted C1-C6 alkyl.

[0136] For example, wherein the A1 (structural unit ) can be selected from:

[0137]

[0138]

[0139] In certain embodiments, wherein said R1, R2 and R3 are each independently selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, When R1, R2 and R3 contain methylene units, the methylene units of said R1, R2 and R3 are each independently not substituted, or the methylene units of said R1, R2 and R3 are each independently substituted by a group selected from the following: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene.

[0140] In certain embodiments, R1 and R2 are independently selected from the group consisting of H, F, Cl, Br and optionally substituted C1-C6 alkyl.

[0141] For example, wherein R1 and R2 can be independently selected from: H, F, Cl, Br and optionally substituted methyl.

[0142] In certain embodiments, wherein said R3 is selected from the group consisting of: hydrogen, optionally substituted -OH, optionally substituted -SH, and optionally substituted C1-C6 alkyl.

[0143] In certain embodiments, when the R3 is an optionally substituted methyl group, the R3 is replaced by R 31 Substituted, the R 31 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 31 When containing a methylene unit, the R 31 The methylene units are each independently not substituted, or the R 31The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0144] In certain embodiments, the R 31 is selected from the group consisting of hydrogen, halogen, optionally substituted -OH, and optionally substituted -SH.

[0145] In certain embodiments, when the R 31 When it is an optionally substituted -OH, the R 31 R 311 Substituted, the R 311 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 311 When containing a methylene unit, the R 311 The methylene units are each independently not substituted, or the R 311 The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0146] In certain embodiments, the R 311 is selected from the group consisting of hydrogen, optionally substituted -P(=O)H2, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted C1-C6 alkyl, and optionally substituted C2-C9 heterocyclylene; wherein, when R 311 When containing a methylene unit, the R 311 The methylene units are each independently not substituted, or the R 311wherein the methylene units are each independently selected from the group consisting of: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H2-, and optionally substituted -NH-; wherein the substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -N + (R)3, -PHR, -P(R)2, -P(=O)R2, -OP(=O)R2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O) OR, -S(O)2NHR, -S(O)2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1- C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy, wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0147] In certain embodiments, the R 311 Selected from the group consisting of hydrogen, optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted

[0148] In certain embodiments, when the R3 is optionally substituted -OH, the R3 is replaced by R 32 Substituted, the R 32is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 32 When containing a methylene unit, the R 32 The methylene units are each independently not substituted, or the R 32 The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0149] In certain embodiments, the R 32 is selected from the group consisting of hydrogen, and optionally substituted C1-C6 alkyl.

[0150] In certain embodiments, when the R 32 When R is an optionally substituted methyl group or an optionally substituted ethyl group, 32 R 321 Substituted, the R 321 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 321 When containing a methylene unit, the R 321 The methylene units are each independently not substituted, or the R 321The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0151] In certain embodiments, the R 321 is selected from the group consisting of hydrogen, halogen, -CN, and optionally substituted C1-C6 alkyl.

[0152] In certain embodiments, the R 321 is selected from the group consisting of hydrogen, F, Cl, -CN, and optionally substituted methyl.

[0153] In certain embodiments, when the R3 is optionally substituted -SH, the R3 is replaced by R 33 Substituted, the R 33 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 33 When containing a methylene unit, the R 33 The methylene units are each independently not substituted, or the R 33 The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0154] In certain embodiments, the R 33 is selected from the group consisting of hydrogen and optionally substituted C1-C6 alkyl.

[0155] In certain embodiments, the R 33 is an optionally substituted methyl group.

[0156] In certain embodiments, when the R 33When it is an optionally substituted methyl group, the R 33 R 331 Substituted, the R 331 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 331 When containing a methylene unit, the R 331 The methylene units are each independently not substituted, or the R 331 The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0157] In certain embodiments, the R 331 Selected from the group consisting of hydrogen, chlorine, fluorine and -CN.

[0158] For example, wherein said R3 can be selected from: optionally substituted -CH2Cl, optionally substituted -CH2SH, optionally substituted -CH2OH, optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted optionally substituted -OH, optionally substituted -OCH3, optionally substituted -OCH2F, optionally substituted -OCH2Cl, optionally substituted -OCH2CN, optionally substituted -OCH2CH3, optionally substituted thiol, optionally substituted -SCH2F, optionally substituted -SCH2Cl, optionally substituted -SCH2CF3, and optionally substituted -SCH2CN.

[0159] In certain embodiments, the compound of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein R1 and R2 are each independently selected from hydrogen, fluorine, chlorine and methyl, and R3 is selected from: -CH2Cl, -CH2SH, -CH2OH, -OCH3, -OCH2F, -OCH2Cl, -OCH2CN, -OCH2CH3, -SH, -SCH2F, -SCH2Cl, -SCH2CF3, and -SCH2CN.

[0160] In certain embodiments, wherein said R1 and R2 are each independently selected from hydrogen, protium, deuterium, tritium and halogen, said R3 is selected from -CH2OH, -SCH2F,

[0161] In certain embodiments, wherein said B is absent or selected from: optionally substituted C1-C6 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C3-C8 heterocyclyl, optionally substituted C6-C 10 Aryl, optionally substituted C5-C 10 heteroaryl, wherein the substituents are selected from the group consisting of protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O)OR, -S(O)2NHR, -S(O)2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl and halogen-substituted C1-C6 alkoxy, wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl and halogen-substituted C1-C6 alkoxy.

[0162] In certain embodiments, wherein said B is selected from: optionally substituted Optionally substituted Optionally substituted X1 is selected from the group consisting of N and optionally substituted CH.

[0163] In certain embodiments, when the X1 is optionally substituted CH, the X1 is substituted by R6, and the R6 is selected from the following group: hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted wherein, when R6 comprises a methylene unit, the methylene units of the R6 are each independently not substituted, or the methylene units of the R6 are each independently selected from the group consisting of: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

[0164] In certain embodiments, said R6 is selected from the group consisting of hydrogen and optionally substituted -OH.

[0165] In certain embodiments, when the R6 is optionally substituted -OH, the R6 is replaced by R 61 Substituted, the R 61 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 61 When containing a methylene unit, the R 61 The methylene units are each independently not substituted, or the R 61 The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0166] In certain embodiments, the R 61 is selected from the group consisting of H and optionally substituted C1-C6 alkyl.

[0167] In certain embodiments, the X1 is selected from the group consisting of CH, C(-O-CH3), and N.

[0168] In certain embodiments, wherein said B is selected from:

[0169] Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted X2 and X3 are each independently selected from the group consisting of O, S and optionally substituted NH.

[0170] In certain embodiments, when X2 or X3 is optionally substituted NH, X2 or X3 is substituted by R7, wherein R7 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted wherein, when R7 comprises a methylene unit, the methylene units of the R7 are each independently not substituted, or the methylene units of the R7 are each independently selected from the group consisting of: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

[0171] In certain embodiments, R7 is selected from the following groups: hydrogen and optionally substituted C1-C6 alkyl; wherein, when R7 contains a methylene unit, the methylene units of R7 are each independently not substituted, or the methylene units of R7 are each independently substituted by: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0172] In certain embodiments, when the R7 is an optionally substituted C1-C6 alkyl, the R7 is replaced by R 71 Substituted, the R 71 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 71 When containing a methylene unit, the R 71 The methylene units are each independently not substituted, or the R 71 The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0173] In certain embodiments, the R 71 is selected from the group consisting of H and optionally substituted C1-C6 alkyl.

[0174] In certain embodiments, said X2 or X3 is selected from the group consisting of NH, N(CH3), O, and S.

[0175] For example, wherein said B can be selected from:

[0176] Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted wherein the substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O)OR, -S(O)2NHR, -S (O)2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy, wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0177] In certain embodiments, the compound of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein B is selected from:

[0178] In certain embodiments, wherein said W is absent or W is selected from: -S(=O)-, -S(=O)2-, -O-, -S-, optionally substituted -NHC(=O)-, optionally substituted -C(=O)NH-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene; wherein, when W comprises an alkylene When the methylene units of W are each independently unsubstituted, or the methylene units of W are each independently substituted by a group selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

[0179] In certain embodiments, when W is an optionally substituted C1-C6 alkylene, W is substituted by R8, and R8 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted wherein, when R8 comprises a methylene unit, the methylene units of the R8 are each independently not substituted, or the methylene units of the R8 are each independently selected from the group consisting of: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted-PH-, optionally substituted-P(=O)H-, optionally substituted-NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

[0180] In certain embodiments, the R8 is selected from the group consisting of hydrogen, =0, optionally substituted C1-C6 alkyl, and optionally substituted cycloalkyl.

[0181] In certain embodiments, said R8 is optionally substituted methyl.

[0182] In certain embodiments, when the R8 is an optionally substituted methyl group, the R8 is replaced by R81 Substituted, the R 81 Selected from the following group: hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl.

[0183] In certain embodiments, the R 81 is selected from the group consisting of halogen and optionally substituted cycloalkyl.

[0184] In certain embodiments, the R 81 is selected from the group consisting of F and optionally substituted cyclopropyl.

[0185] In certain embodiments, wherein W is absent or W is selected from: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -CH2NHC(=O)-, optionally substituted -NHC(=O)CH2-, optionally substituted -C(=O)NH-, optionally substituted -NH-, optionally substituted -CH=CH-, -C≡C-, optionally substituted C1-C6 alkylene, optionally substituted -OCH2-, optionally substituted -CH2O-, optionally substituted -SCH2-, optionally substituted -CH2S-, optionally substituted -NHC(=O)-, optionally substituted -C(=O)CH2-, optionally substituted -CH2NH-, optionally substituted -NHCH2-, optionally substituted Optionally substituted Optionally substituted Optionally substituted and optionally substituted

[0186] For example, the W may be absent or W may be selected from:

[0187]

[0188] In certain embodiments, the compound of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein W is absent or W is

[0189] For another example, wherein said B may not exist or be selected from:

[0190] Said W may not exist or W is selected from:

[0191] Said A1 may be selected from:

[0192]

[0193] In certain embodiments, the compound of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, is selected from

[0194]

[0195] in,

[0196] Said X is selected from -O-, -S- and -NH-;

[0197] R4 and R5 are each independently selected from the group consisting of H, F, Cl, -OH, -NH2, and C1-C6 alkyl;

[0198] Said n is selected from 1, 2 or 3;

[0199] Said Y1 is absent or selected from: hydroxyl, thiol, amino and C1-C6 alkyl;

[0200] Said R1 and R2 are each independently selected from hydrogen, fluorine, chlorine and methyl;

[0201] The R3 is selected from: -CH2Cl, -CH2SH, -CH2OH, -OCH3, -OCH2F, -OCH2Cl, -OCH2CN, -OCH2CH3, -SH, -SCH2F, -SCH2Cl, -SCH2CF3, and -SCH2CN.

[0202] For example, the compound of formula I can be selected from the following structures:

[0203]

[0204]

[0205]

[0206]

[0207]

[0208]

[0209]

[0210]

[0211]

[0212]

[0213]

[0214]

[0215]

[0216]

[0217]

[0218]

[0219]

[0220]

[0221]

[0222]

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231]

[0232]

[0233]

[0234]

[0235]

[0236]

[0237]

[0238]

[0239]

[0240]

[0241]

[0242]

[0243]

[0244]

[0245]

[0246]

[0247]

[0248]

[0249]

[0250]

[0251]

[0252]

[0253]

[0254]

[0255]

[0256]

[0257]

[0258]

[0259]

[0260]

[0261]

[0262]

[0263]

[0264]

[0265]

[0266]

[0267]

[0268]

[0269]

[0270] In another aspect, the present invention provides a compound represented by the following formula or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

[0271]

[0272] In another aspect, the present application provides a compound of formula IIa or IIb:

[0273]

[0274] or its tautomers, mesomers, racemates, enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, wherein,

[0275] R1, R2, R3, R4 and R5 are each independently any group;

[0276] B does not exist or B is any group;

[0277] W does not exist or W is any group;

[0278] A2 is a substituted benzene ring;

[0279] Each CR4R5 unit is independently unsubstituted or substituted by a member selected from the group consisting of -O-, -S-, -NR-, -S(O)-, -S(O)2-, -C(=O)-, -C=C-, and -C≡C-, wherein R4 and R5 may together form an optionally substituted cycloalkyl, an optionally substituted heterocyclyl, an optionally substituted bridged ring group, an optionally substituted heterobridged ring group, an optionally substituted spirocyclyl, and an optionally substituted heterospirocyclyl, and n is any integer from 1 to 20;

[0280] X is selected from: -O-, -S-, -NR-;

[0281] Y1 is any group, m is any integer from 0 to 4;

[0282] Y2 is selected from -O-, -S- and -NR-;

[0283] wherein the substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O)OR, -S(O)2NHR, -S( O)2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy; wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0284] In certain embodiments, the wavy line in Formula IIa or IIb It indicates that the ligand is directly connected to the ligand through the X or Y2 group, or is connected to the ligand through a linker fragment.

[0285] In certain preferred embodiments of the present invention, certain groups in the compounds of formula (IIa) and (IIb) or their pharmaceutically acceptable salts are defined as follows, and the unmentioned groups are the same as those described in any scheme of the present application (referred to as "in certain embodiments").

[0286] In certain embodiments, each of the CR4R5 units is independently unsubstituted or substituted with a member selected from the group consisting of -O-, -S-, and -C(=O)-.

[0287] In certain embodiments, wherein said R4 and R5 together form an optionally substituted cycloalkyl or an optionally substituted heterocyclic radical. For example, said R4 and R5 together can form a cyclopropyl or cyclobutyl.

[0288] In certain embodiments, wherein said R4 and R5 are each independently selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; Wherein, when R4 and R5 contain methylene units, the methylene units of R4 and R5 are each independently not substituted, or the methylene units of R4 and R5 are each independently substituted by a group selected from the following groups: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene.

[0289] In certain embodiments, wherein said R4 and R5 are each independently selected from the following group: hydrogen, protium, deuterium, tritium, halogen, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R4 and R5 contain methylene units, said methylene units of said R4 and R5 are Each of the methylene units of R4 and R5 is independently not substituted, or each of the methylene units of R4 and R5 is independently substituted by a group selected from the following groups: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene.

[0290] In certain embodiments, the compound of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein R4 and R5 are each independently selected from the following groups: H, F, Cl, -OH, -NH2 and C1-C6 alkyl; and n is selected from 1, 2 or 3.

[0291] In certain embodiments, wherein said Y1 is selected from: protium, deuterium, tritium, halogen, -OR, -SR, -NHR, -N(R)2, -PHR, -P(R)2, -P(=O)HR, -P(=O)R2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0292] In certain embodiments, wherein said Y1 is selected from: protium, deuterium, tritium, halogen, -OR, -SR, -NHR, -N(R)2, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy; wherein each R is independently selected from hydrogen, protium, deuterium, tritium, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0293] In certain embodiments, Y1 is absent or selected from: -OR, -SR, -N(R)2 and optionally substituted C1-C6 alkyl; wherein each R is independently selected from hydrogen, protium, deuterium, tritium, C1-C6 alkyl, C1-C6 alkoxy.

[0294] In certain embodiments, the Y1 is absent or selected from the group consisting of: hydroxyl, thiol, amino, and C1-C6 alkyl.

[0295] In certain embodiments, Y2 comprises -NR-; wherein each R is independently selected from hydrogen, protium, deuterium, tritium, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0296] For example, the A2 (structural unit ) can be selected from:

[0297]

[0298]

[0299] In certain embodiments, wherein said R1, R2 and R3 are each independently selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, When R1, R2 and R3 contain methylene units, the methylene units of said R1, R2 and R3 are each independently not substituted, or the methylene units of said R1, R2 and R3 are each independently substituted by a group selected from the following: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene and optionally substituted heteroarylene.

[0300] In certain embodiments, R1 and R2 are independently selected from the group consisting of H, F, Cl, Br and optionally substituted C1-C6 alkyl.

[0301] In certain embodiments, R1 and R2 are independently selected from the group consisting of H, F, Cl, Br and optionally substituted methyl.

[0302] In certain embodiments, wherein said R3 is selected from the group consisting of: hydrogen, optionally substituted -OH, optionally substituted -SH, and optionally substituted C1-C6 alkyl.

[0303] In certain embodiments, when the R3 is an optionally substituted methyl group, the R3 is replaced by R 31 Substituted, the R 31 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 31 When containing a methylene unit, the R 31 The methylene units are each independently not substituted, or the R 31The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0304] In certain embodiments, the R 31 is selected from the group consisting of hydrogen, halogen, optionally substituted -OH, and optionally substituted -SH.

[0305] In certain embodiments, when the R 31 When it is an optionally substituted -OH, the R 31 R 311 Substituted, the R 311 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 311 When containing a methylene unit, the R 311 The methylene units are each independently not substituted, or the R 311 The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0306] In certain embodiments, the R 311 is selected from the group consisting of hydrogen, optionally substituted -P(=O)H2, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted C1-C6 alkyl, and optionally substituted C2-C9 heterocyclylene; wherein, when R 311 When containing a methylene unit, the R 311 The methylene units are each independently not substituted, or the R 311The methylene units are each independently selected from the group consisting of: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H2-, and optionally substituted -NH-; wherein the substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -N + (R)3, -PHR, -P(R)2, -P(=O)R2, -OP(=O)R2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O) OR, -S(O)2NHR, -S(O)2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1- C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy, wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0307] In certain embodiments, the R 311 Selected from the group consisting of hydrogen, optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted

[0308] In certain embodiments, when the R3 is optionally substituted -OH, the R3 is replaced by R 32 Substituted, the R 32is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 32 When containing a methylene unit, the R 32 The methylene units are each independently not substituted, or the R 32 The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0309] In certain embodiments, the R 32 is selected from the group consisting of hydrogen, and optionally substituted C1-C6 alkyl.

[0310] In certain embodiments, when the R 32 When R is an optionally substituted methyl group or an optionally substituted ethyl group, 32 R 321 Substituted, the R 321 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 321 When containing a methylene unit, the R 321 The methylene units are each independently not substituted, or the R 321The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0311] In certain embodiments, the R 321 is selected from the group consisting of hydrogen, halogen, -CN, and optionally substituted C1-C6 alkyl.

[0312] In certain embodiments, the R 321 is selected from the group consisting of hydrogen, F, Cl, -CN, and optionally substituted methyl.

[0313] In certain embodiments, when the R3 is optionally substituted -SH, the R3 is replaced by R 33 Substituted, the R 33 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 33 When containing a methylene unit, the R 33 The methylene units are each independently not substituted, or the R 33 The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0314] In certain embodiments, the R 33 is selected from the group consisting of hydrogen, and optionally substituted C1-C6 alkyl.

[0315] In certain embodiments, when the R 33 When R is an optionally substituted methyl group or an optionally substituted ethyl group, 33 R 331 Substituted, the R331 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 331 When containing a methylene unit, the R 331 The methylene units are each independently not substituted, or the R 331 The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0316] In certain embodiments, the R331 is selected from the group consisting of hydrogen, chlorine, fluorine, and -CN.

[0317] For example, wherein said R3 can be selected from: optionally substituted -CH2Cl, optionally substituted -CH2SH, optionally substituted -CH2OH, optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted optionally substituted -OH, optionally substituted -OCH3, optionally substituted -OCH2F, optionally substituted -OCH2Cl, optionally substituted -OCH2CN, optionally substituted -OCH2CH3, optionally substituted thiol, optionally substituted -SCH2F, optionally substituted -SCH2Cl, optionally substituted -SCH2CF3, and optionally substituted -SCH2CN.

[0318] In certain embodiments, the compound of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein R3 is selected from: -CH2Cl, -CH2SH, -CH2OH, -OCH3, -OCH2F, -OCH2Cl, -OCH2CN, -OCH2CH3, -SH, -SCH2F, -SCH2Cl, -SCH2CF3, and -SCH2CN.

[0319] In certain embodiments, wherein said R1 and R2 are each independently selected from hydrogen, protium, deuterium, tritium and halogen, said R3 is selected from -CH2-OH, -SCH2F,

[0320] In certain embodiments, wherein said B is absent or selected from: optionally substituted C1-C6 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C3-C8 heterocyclyl, optionally substituted C6-C 10 Aryl, optionally substituted C5-C 10 heteroaryl, wherein the substituents are selected from the group consisting of protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O)OR, -S(O)2NHR, -S(O)2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy, wherein each R is independently selected from hydrogen, protium, deuterium, tritium, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0321] In certain embodiments, wherein said B is selected from: optionally substituted Optionally substituted Optionally substituted X1 is selected from the group consisting of N and optionally substituted CH.

[0322] In certain embodiments, when the X1 is optionally substituted CH, the X1 is substituted by R6, and the R6 is selected from the following group: hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted wherein, when R6 comprises a methylene unit, the methylene units of the R6 are each independently not substituted, or the methylene units of the R6 are each independently selected from the group consisting of: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

[0323] In certain embodiments, said R6 is selected from the group consisting of hydrogen and optionally substituted -OH.

[0324] In certain embodiments, when the R6 is optionally substituted -OH, the R6 is replaced by R 61 Substituted, the R 61 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 61 When containing a methylene unit, the R 61 The methylene units are each independently not substituted, or the R 61 The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0325] In certain embodiments, the R 61 is selected from the group consisting of H and optionally substituted C1-C6 alkyl.

[0326] In certain embodiments, X1 is selected from the group consisting of CH, C(-O-CH3) and N.

[0327] In certain embodiments, wherein said B can be selected from: optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted X2 and X3 are each independently selected from the group consisting of O, S and optionally substituted NH.

[0328] In certain embodiments, when X2 or X3 is optionally substituted NH, X2 or X3 is substituted by R7, wherein R7 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted wherein, when R7 comprises a methylene unit, the methylene units of the R7 are each independently not substituted, or the methylene units of the R7 are each independently selected from the group consisting of: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

[0329] In certain embodiments, the R7 is selected from the group consisting of hydrogen and optionally substituted C1-C6 alkyl.

[0330] In certain embodiments, when the R7 is an optionally substituted C1-C6 alkyl, the R7 is replaced by R71 Substituted, the R 71 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein, when R 71 When containing a methylene unit, the R 71 The methylene units are each independently not substituted, or the R 71 The methylene units are each independently selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0331] In certain embodiments, the R 71 is selected from the group consisting of H and optionally substituted C1-C6 alkyl.

[0332] In certain embodiments, said X2 or X3 is selected from the group consisting of NH, N(CH3), O, and S.

[0333] In certain embodiments, wherein said B is selected from:

[0334] Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted wherein the substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O)OR, -S(O)2NHR, -S( O)2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy, wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0335] In certain embodiments, the compound of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein B is selected from:

[0336] In certain embodiments, wherein W is absent or W is selected from: -S(=O)-, -S(=O)2-, -O-, -S-, -NHC(=O)-, -C(=O)NH-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted arylene, and optionally substituted heteroarylene; wherein, when W comprises a methylene unit , the methylene units of the W are each independently unsubstituted, or the methylene units of the W are each independently substituted by a group selected from the group consisting of -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene, and optionally substituted heteroarylene.

[0337] In certain embodiments, when W is an optionally substituted C1-C6 alkylene, W is substituted by R8, wherein R8 is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH 2. optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl; wherein, when R8 contains a methylene unit, the methylene units of the R8 are each independently not substituted, or the methylene units of the R8 are each independently selected from the following groups: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -PH-, optionally substituted -P(=O)H-, optionally substituted -NH-, optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted cycloalkylene, optionally substituted heterocyclylene, optionally substituted arylene and optionally substituted heteroarylene.

[0338] In certain embodiments, the R8 is selected from the group consisting of hydrogen, =0, optionally substituted C1-C6 alkyl, and optionally substituted cycloalkyl.

[0339] In certain embodiments, said R8 is optionally substituted methyl.

[0340] In certain embodiments, when the R8 is an optionally substituted methyl group, the R8 is replaced by R 81 Substituted, the R 81 Selected from the following group: hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, =O, =S, optionally substituted -S(=O)H, optionally substituted -S(=O)2H, optionally substituted -C(=O)H, optionally substituted -OH, optionally substituted -SH, optionally substituted -PH2, optionally substituted -P(=O)H2, optionally substituted -NH2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and optionally substituted heteroaryl.

[0341] In certain embodiments, the R 81 is selected from the group consisting of halogen and optionally substituted cycloalkyl.

[0342] In certain embodiments, the R 81 is selected from the group consisting of F and optionally substituted cyclopropyl.

[0343] In certain embodiments, wherein W is absent or W is selected from: -S(=O)-, -S(=O)2-, -O-, -S-, -C(=O)-, optionally substituted -CH2NHC(=O)-, optionally substituted -NHC(=O)CH2-, optionally substituted -C(=O)NH-, optionally substituted -NH-, optionally substituted -CH=CH-, -C≡C-, optionally substituted C1-C6 alkylene, optionally substituted -OCH2-, optionally substituted -CH2O-, optionally substituted -SCH2-, optionally substituted -CH2S-, optionally substituted -NHC(=O)-, optionally substituted -COCH2-, optionally substituted -CH2NH-, optionally substituted -NHCH2-, optionally substituted -CH(CH3)-, optionally substituted Optionally substituted Optionally substituted Optionally substituted and optionally substituted

[0344] For example, wherein said W may be absent or W is selected from:

[0345]

[0346] In certain embodiments, the compound of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein W is absent or W is

[0347] For example, said B may be absent or selected from:

[0348] Said W may not exist or W is selected from:

[0349] Said A2 can be selected from:

[0350]

[0351]

[0352] In certain embodiments, the compound of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, is selected from

[0353]

[0354]

[0355] in,

[0356] Said X is selected from -O-, -S- and -NH-;

[0357] R4 and R5 are each independently selected from the group consisting of H, F, Cl, -OH, -NH2, and C1-C6 alkyl;

[0358] Said n is selected from 1, 2 or 3;

[0359] Said Y1 is absent or selected from: hydroxyl, thiol, amino and C1-C6 alkyl;

[0360] Said R1 and R2 are each independently selected from hydrogen, fluorine, chlorine and methyl;

[0361] The R3 is selected from: -CH2Cl, -CH2SH, -CH2OH, -OCH3, -OCH2F, -OCH2Cl, -OCH2CN, -OCH2CH3, -SH, -SCH2F, -SCH2Cl, -SCH2CF3, and -SCH2CN.

[0362] In certain embodiments, the compound of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

[0363]

[0364] In certain embodiments, the compound of Formula IIa or IIb, or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, further comprises a Trigger fragment (Tr), and the compound comprises the following structure:

[0365]

[0366] Wherein said Tr is selected from the following structures:

[0367]

[0368] R1, R2, R3, R4 and R5 are each independently any group;

[0369] B does not exist or B is any group;

[0370] W does not exist or W is any group;

[0371] Each CR4R5 unit is independently unsubstituted or substituted by a member selected from the group consisting of -O-, -S-, -NR-, -S(O)-, -S(O)2-, -C(=O)-, -C=C-, and -C≡C-, wherein R4 and R5 may together form an optionally substituted cycloalkyl, an optionally substituted heterocyclyl, an optionally substituted bridged ring group, an optionally substituted heterobridged ring group, an optionally substituted spirocyclyl, and an optionally substituted heterospirocyclyl, and n is any integer from 1 to 20;

[0372] X is selected from: -O-, -S-, -NR-;

[0373] Y1 is any group, m is any integer from 0 to 4;

[0374] Y2 is selected from -O-, -S- and -NR-;

[0375] wherein the substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O)OR, -S(O)2NHR, -S( O)2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy; wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0376] For example, wherein R1 and R2 can be independently selected from: H, F, Cl, Br and optionally substituted methyl.

[0377] For example, wherein said R3 can be selected from: optionally substituted -CH2Cl, optionally substituted -CH2SH, optionally substituted -CH2OH, optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted optionally substituted -OH, optionally substituted -OCH3, optionally substituted -OCH2F, optionally substituted -OCH2Cl, optionally substituted -OCH2CN, optionally substituted -OCH2CH3, optionally substituted thiol, optionally substituted -SCH2F, optionally substituted -SCH2Cl, optionally substituted -SCH2CF3, and optionally substituted -SCH2CN.

[0378] For example, wherein said B can be selected from:

[0379] Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted wherein the substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O)OR, -S(O)2NHR, -S (O)2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy, wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0380] For example, the W may be absent or W may be selected from:

[0381]

[0382] In certain embodiments, the linker fragment includes an L1 fragment, an L2 fragment, and / or an L3 fragment, and the compound has the following structure:

[0383]

[0384] in,

[0385] Tr does not exist or Tr is any group;

[0386] L3 is selected from a polypeptide fragment;

[0387] L2 is absent or selected from a connecting fragment;

[0388] L1 is selected from a coupling unit;

[0389] R1, R2, R3, R4 and R5 are each independently any group;

[0390] B does not exist or B is any group;

[0391] W does not exist or W is any group;

[0392] Each CR4R5 unit is independently unsubstituted or substituted by a member selected from the group consisting of -O-, -S-, -NR-, -S(O)-, -S(O)2-, -C(=O)-, -C=C-, and -C≡C-, wherein R4 and R5 may together form an optionally substituted cycloalkyl, an optionally substituted heterocyclyl, an optionally substituted bridged ring group, an optionally substituted heterobridged ring group, an optionally substituted spirocyclyl, and an optionally substituted heterospirocyclyl, and n is any integer from 1 to 20;

[0393] X is selected from: -O-, -S-, -NR-;

[0394] Y1 is any group, m is any integer from 0 to 4;

[0395] Y2 is selected from -O-, -S- and -NR-;

[0396] wherein the substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O)OR, -S(O)2NHR, -S( O)2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy; wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0397] In certain preferred embodiments of the present invention, certain groups in the compounds of formula (IVa) and (IVb) or their pharmaceutically acceptable salts are defined as follows, and the unmentioned groups are the same as those described in any embodiment of the present application (referred to as "in certain embodiments").

[0398] In certain embodiments, the L3 is selected from a dipeptide, a tripeptide, and a tetrapeptide.

[0399] In certain embodiments, the dipeptide is selected from the group consisting of: GA, GG, AG, EG, EA, GE, DG, DA, GD, VC, VA, AA, and VK.

[0400] In certain embodiments, the tripeptide is selected from the group consisting of: EAG, EGG, GEG, GEA, DAG, DGG, GDG, GDA, GGA, GAG, GFG, AAG, AAA, VAG, VCG, VKG.

[0401] In certain embodiments, the tetrapeptide is selected from the group consisting of: GGFG, GGAG, GGGG, GEGG, GEAG, GDGG, GDAG, AAAG, and EAGG.

[0402] In certain embodiments, the L3 is selected from the following groups: glycine-glycine-phenylalanine-glycine (GGFG), alanine-alanine-alanine-glycine (AAAG), glycine-glycine-glycine-glycine (GGGG), valine-alanine-glycine (VAG), valine-citrulline-glycine (VCG), alanine-alanine-glycine (AAG), alanine-alanine-alanine (AAA), valine-alanine (VA), valine-citrulline (VC), alanine-alanine-glycine (AAG), alanine-alanine-alanine (AAA), valine-alanine (VA), valine-citrulline (VC), alanine The amino acids glutamate-alanine (AA), glutamate-alanine-glycine-glycine (EAGG), glycine-glutamate-alanine-glycine (GEAG), glycine-glutamate-glycine-glycine (GEGG), glutamate-glycine-glycine (EGG), glutamate-alanine-glycine (EAG), valine-lysine-glycine (VKG), glycine-glutamate-glycine (GEG), glutamate-alanine (EA), glutamate-glycine (EG) and glycine-glutamate (GE).

[0403] In certain embodiments, wherein said L2 is absent.

[0404] In certain embodiments, the L2 comprises or does not comprise a PEG branched chain or a PEG linear chain.

[0405] In certain embodiments, when the L2 does not comprise PEG, the L2 is selected from:

[0406]

[0407] In certain embodiments, when the L2 comprises a PEG linear chain, the L2 is selected from:

[0408]

[0409] Wherein p is any integer from 1 to 20.

[0410] In certain embodiments, when the L2 comprises a PEG linear chain, the L2 is selected from:

[0411]

[0412] In certain embodiments, when the L2 comprises a PEG side chain, the L2 is selected from:

[0413]

[0414]

[0415] wherein q is selected from any integer between 1 and 30.

[0416] In certain embodiments, when the L2 comprises a PEG side chain, the L2 is selected from:

[0417]

[0418]

[0419]

[0420]

[0421] In certain embodiments, when L1 is coupled to a sulfhydryl group of a ligand, L1 is selected from:

[0422] Among them, the R L1a 、R L1b 、R L1c Each is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, alkyl, alkenyl, alkynyl, alicyclic, heterocyclic, aryl and heteroaryl.

[0423] In certain embodiments, the R L1a 、R L1b 、R L1c Each is independently selected from the group consisting of hydrogen, optionally substituted methyl, optionally substituted ethyl, optionally substituted aryl, and optionally substituted benzyl.

[0424] In certain embodiments, when L1 is coupled to the ligand via an amino group, the L1 is selected from:

[0425]

[0426] In certain embodiments, when L1 is coupled via click chemistry, the L1 is selected from:

[0427]

[0428] For example, the compound of formula IVa or IVb can be selected from the following structures:

[0429]

[0430]

[0431]

[0432]

[0433]

[0434]

[0435]

[0436]

[0437]

[0438]

[0439]

[0440]

[0441]

[0442]

[0443]

[0444]

[0445]

[0446]

[0447]

[0448]

[0449]

[0450]

[0451] In certain embodiments, the compound of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein the compound of formula IVa or IVb is selected from the following structures:

[0452]

[0453]

[0454] In certain embodiments, the compound of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein the compound further comprises a linker fragment, the compound of Formula IIa or IIb can be coupled to a ligand via the linker fragment, and the compound has the following structure:

[0455]

[0456] in,

[0457] Tr does not exist or Tr is any group;

[0458] L3 is selected from a polypeptide fragment;

[0459] L2 is absent or selected from a connecting fragment;

[0460] L1 is selected from a coupling unit; in the general formula IVa-1 and IVb-1, L1 is a connecting form;

[0461] R1, R2, R3, R4, R5, B, W, CR4R5, n, X, Y1 and Y2 are as described in any one of the present invention;

[0462] Wavy lines in the general formula Indicates that it is connected to the ligand through the L1 group.

[0463] In certain embodiments, the compound of the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein the structural unit -Tr-L3-L2-L1- is selected from:

[0464]

[0465] On the other hand, the present application provides a conjugate comprising the aforementioned compound or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof.

[0466] In certain embodiments, it comprises a ligand-drug conjugate. For example, the conjugate may comprise an antibody-drug conjugate.

[0467] In certain embodiments, the ligand comprises an antibody or an antigen-binding fragment thereof.

[0468] In certain embodiments, the antibody is selected from the group consisting of: a human antibody, a humanized antibody, a chimeric antibody, a multispecific antibody, a monoclonal antibody, and a polyclonal antibody.

[0469] In certain embodiments, the antigen-binding fragment is selected from the group consisting of: Fab, Fab', F(ab')2, Fv, scFv, diabody, Fd, dAb, VHH, big antibody and complementarity determining region (CDR) fragment.

[0470] In certain embodiments, the ligand specifically binds to an antigen selected from the group consisting of AXL, BAFFR, BCMA, BCR-list components, BDCA2, BDCA4, BTLA, BTNL2BTNL3, BTNL8, BTNL9, C10orf54, CCR1, CCR3, CCR4, CCR5, CCR6, CCR7, CCR9, CCR10, CD11c, CD137, CD138, CD14, CD163, CD168, CD 177, CD19, CD20, CD209, CD209L, CD22, CD226, CD248, CD25, CD27, CD274, CD276, CD28, CD30, CD300A, CD33, CD37, CD38, CD4, differentiation antigen cluster 40 (C D40), CD44, CD45, CD46, CD47, CD48, CD5, CD52, CD55, CD56, CD59, CD62E, CD68, CD69, CD70, CD74, CD79a, CD79b, CD8, CD80, CD86, CD90.2. CD96, CLEC12A, CLEC12B, CLEC7A, CLEC9A, CR1, CR3, CRTAM, CSF1R, CTLA4, CXCR1 / 2, CXCR4, C XCR5, DDR1, DDR2, DEC-205, DLL4, DR6, FAP, FCamR, FCMR, FcR's, Fire, GITR, HHLA2, HLA type II (HLA class II), HVEM, ICOSLG, IFNAR, type I interferon receptor subunit (IFNAR1), IFNLR1, IL10R1, IL10R2, IL12R, IL13RA1, IL13RA2, IL15R, IL17RA, IL17RB, IL17RC, IL17RE, IL20R1, IL20R2, IL21R, IL22R1, IL22RA, IL23R, IL27R, IL29R, IL2Rg, IL31R, IL36R, IL3RA, IL4R, IL6R, IL5R, IL7R, IL9R, Integrins, LAG3, LIFR, MAG / Siglec-4 (sialic acid-binding immunoglobulin-like lectin-4), MMR, MSR1, NCR3LG1, NKG2D, NKp30, NKp46, OX40 (CD134), PDCD1, PROKR1, PVR, PVRIG, PVRL2, PVRL3, RELT, SIGIRR, Siglec-1 (sialic acid-binding immunoglobulin-like lectin-1), Siglec-10, Siglec-5, Siglec-6, Siglec-7, Siglec-8, Siglec-9, SIRPA, SLAMF7, TACI, TCR-listcomponents / assoc, PTCRA, TCRb, CD3z, CD3, TEK, TGFBR1, TGFBR2, TGFBR3, TIGIT, TLR2, TLR4, tumor necrosis factor alpha (TNFα), TROY, TSLPR, TYRO, VLDLR, VSIG4, IL2R-y, and VTCN1.

[0471] In certain embodiments, the ligand specifically binds to: TNFα, CD40, and / or IFNAR1.

[0472] In certain embodiments, the ligand is selected from the group consisting of: an anti-TNFα antibody or an antigen-binding fragment thereof, an anti-CD40 antibody or an antigen-binding fragment thereof, and an anti-IFNAR1 antibody or an antigen-binding fragment thereof. In certain embodiments, the ligand is selected from the group consisting of: an anti-TNFα antibody or an antigen-binding fragment thereof, an anti-CD40 antibody or an antigen-binding fragment thereof, an anti-BDCA2 antibody or an antigen-binding fragment thereof, and an anti-IFNAR1 antibody or an antigen-binding fragment thereof.

[0473] For example, the ligand can be selected from: anti-TNFα monoclonal antibody, anti-CD40 monoclonal antibody, and anti-IFNAR1 monoclonal antibody.

[0474] The present disclosure also provides immunoconjugates containing glucocorticoid receptor agonists connected to anti-TNFα proteins. In certain embodiments, the anti-TNFα protein is an antibody or its antigen-binding fragment. In certain embodiments, the anti-TNFα protein is an antibody or its antigen-binding fragment in conjunction with TNFα (e.g., soluble TNFα and / or membrane-bound TNFα). In certain embodiments, the anti-TNFα protein is a soluble TNF receptor protein, such as a soluble TNF receptor protein fused to a heavy chain constant domain or its fragment (such as Fc). In certain embodiments, the anti-TNFα protein (e.g., an anti-TNF antibody, its antigen-binding fragment or a soluble TNF receptor) can be combined with TNFα on the cell surface and become internalized. For example, US2014 / 0294813 (which is incorporated herein by reference in its entirety) discloses an anti-TNF protein that displays internalization after being combined with cell surface human TNF. In certain embodiments, an antibody or its antigen-binding fragment is combined with human and / or mouse TNF-α. Antibodies and antigen-binding fragments in conjunction with TNF-α are known in the art.

[0475] Anti-TNF-α antibodies and antigen-binding fragments thereof include, for example, adalimumab, infliximab, certolizumab pegol, afelimomab, nerelimomab, ozoralizumab, placulumab, golimumab, and anti-murine TNFαmIgG2a. Additional anti-TNF-α antibodies and antigen-binding fragments are provided in, for example, WO 2013 / 087912, WO 2014 / 152247, and WO 2015 / 073884, each of which is incorporated herein by reference in its entirety.

[0476] Adalimumab is described in U.S. Patent No. 6,258,562, which is incorporated herein by reference in its entirety. Infliximab is described in U.S. Patent No. 5,656,272, which is incorporated herein by reference in its entirety. Certolizumab is discussed in WO 01 / 94585, which is incorporated herein by reference in its entirety. Afelimomab (also known as MAK195) is discussed in Vincent, Int. J. Clin. Pract. 54: 190-193 (2000), which is incorporated herein by reference in its entirety. Ozolanzumab (also known as ATN-103) is a nanobody. It contains three heavy chain variable regions fused by a GlySer linker. Variable regions 1 and 3 are identical, and ozolanzumab does not contain heavy chains. Ozolanzumab is discussed in WO 2012 / 131053, which is incorporated herein by reference in its entirety. Prakritumab (also known as CEP-37247) is a domain antibody composed of a dimer of VL-pCH1-CH2-CH3 or [V-κ]2-Fc and is discussed in Gay et al., Mabs 2: 625-638 (2010), which is incorporated herein by reference in its entirety. Golimumab (also known as CNTO 148) is discussed in WO2013 / 087912 and the sequences are provided in GenBank: DI496971.1 and GenBank DI 496970.1, each of which is incorporated herein by reference in its entirety. Anti-murine TNFαmIgG2a is described in the literature: McRae BL et al. J Crohns Colitis 10(1): 69-76 (2016), each of which is incorporated herein by reference in its entirety.

[0477] Anti-TNF-α antibodies and antigen-binding fragments thereof also include antibodies and antigen-binding fragments thereof that competitively inhibit the binding of adalimumab, infliximab, certolizumab plucatum, afelimomab, nerimumab, ozolazumab, prukumab, or golimumab to TNF-α. Anti-TNF-α antibodies and antigen-binding fragments thereof also include antibodies and antigen-binding fragments that bind to the same TNF-α epitope as adalimumab, infliximab, certolizumab plucatum, afelimomab, nerimumab, ozolazumab, prukumab, or golimumab.

[0478] In certain embodiments, the anti-TNF-α antibody or antigen-binding fragment thereof competitively inhibits the binding of adalimumab to TNF-α. In certain embodiments, the anti-TNF-α antibody or antigen-binding fragment thereof binds to the same TNF-α epitope as adalimumab. In certain embodiments, the anti-TNF-α antibody or antigen-binding fragment thereof is adalimumab or an antigen-binding fragment thereof. In certain embodiments, the anti-TNF-α antibody or antigen-binding fragment thereof is adalimumab.

[0479] In certain embodiments, the anti-TNF-α antibody or antigen-binding fragment thereof comprises a sequence of adalimumab, infliximab, certolizumab plucatumumab, afelimomab, nerimumab, ozolazumab, prukumab, or golimumab, such as the complementarity determining regions (CDRs), variable heavy chain domain (VH), and / or variable light chain domain (VL).

[0480] In certain embodiments, the anti-TNFα antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) of an antibody, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, each of which has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the HCDR1, HCDR2, and HCDR3 of the following molecules, respectively: adalimumab, infliximab, afelimomab, or golimumab.

[0481] In certain embodiments, the anti-TNFα antibody or antigen-binding fragment thereof comprises a heavy chain variable region of an antibody, wherein the heavy chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the heavy chain variable region of the following molecules, respectively: Adalimumab, Infliximab, Afelimomab, or Golimumab.

[0482] In certain embodiments, the anti-TNFα antibody or antigen-binding fragment thereof comprises a heavy chain of an antibody, wherein the heavy chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the heavy chain of adalimumab, infliximab, afelimomab, or golimumab, respectively.

[0483] In certain embodiments, the anti-TNFα antibody or antigen-binding fragment thereof comprises a light chain variable region (VL) of an antibody, wherein the light chain variable region comprises LCDR1, LCDR2, and LCDR3, each of which has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the LCDR1, LCDR2, and LCDR3 of the following molecules, respectively: adalimumab, infliximab, afelimomab, or golimumab.

[0484] In certain embodiments, the anti-TNFα antibody or antigen-binding fragment thereof comprises a light chain variable region of an antibody, wherein the light chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the light chain variable region of the following molecules, respectively: Adalimumab, Infliximab, Afelimomab, or Golimumab.

[0485] In certain embodiments, the anti-TNFα antibody or antigen-binding fragment thereof comprises a light chain of an antibody, wherein the light chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the light chain of adalimumab, infliximab, afelimomab, or golimumab, respectively.

[0486] In certain embodiments, the present application provides an antibody or antigen-binding fragment thereof that can specifically bind to TNF-α and comprises the Chothia VL CDRs of the VL of adalimumab, infliximab, certolizumab plumol, afelimomab, nerimumab, ozolazumab, prukumab, or golimumab. In certain aspects, the present application provides an antibody or antigen-binding fragment thereof that specifically binds to TNF-α and comprises the Chothia VH CDRs of the VH of adalimumab, infliximab, certolizumab plumol, afelimomab, nerimumab, ozolazumab, prukumab, or golimumab. In certain aspects, provided herein are antibodies or antigen-binding fragments thereof that specifically bind to TNF-α and comprise the Chothia VL CDRs of the VL of adalimumab, infliximab, certolizumab pruzimab, afelimomab, nerimumab, ozolazumab, pruzilumab, or golimumab and comprise the Chothia VH CDRs of the VH of adalimumab, infliximab, certolizumab pruzilumab, afelimomab, nerimumab, ozolazumab, pruzilumab, or golimumab.

[0487] In certain embodiments, the anti-TNFα antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) of an antibody, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, each of which has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity with the HCDR1, HCDR2, and HCDR3 of the following molecules, respectively: Adalimumab, Infliximab, Afelimomab, Golimumab, or anti-murine TNFαmIgG2a 8c11.

[0488] In certain embodiments, the anti-TNFα antibody or antigen-binding fragment thereof comprises a heavy chain variable region of an antibody, wherein the heavy chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity with the heavy chain variable region of the following molecules, respectively: Adalimumab, Infliximab, Afelimomab, golimumab, or anti-murine TNFαmIgG2a8c11.

[0489] In certain embodiments, the anti-TNFα antibody or antigen-binding fragment thereof comprises a heavy chain of an antibody, wherein the heavy chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the heavy chain of the following molecules, respectively: Adalimumab, Infliximab, Afelimomab, golimumab, or anti-murine TNFαmIgG2a 8c11.

[0490] In certain embodiments, the anti-TNFα antibody or antigen-binding fragment thereof comprises a light chain variable region (VL) of an antibody, wherein the light chain variable region comprises LCDR1, LCDR2, and LCDR3, each of which has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity with the LCDR1, LCDR2, and LCDR3 of the following molecules, respectively: adalimumab, infliximab, afelimomab, golimumab, or anti-murine TNFαmIgG2a 8c11.

[0491] For example, the anti-TNFα antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL) of an antibody, wherein the heavy chain variable region comprises HCDR1, HCDR2, HCDR3, and the light chain variable region comprises LCDR1, LCDR2, LCDR3, each of which has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity with the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of the following molecules, respectively: Adalimumab, Infliximab, Afelimomab, Golimumab, or anti-murine TNFαmIgG2a 8c11.

[0492] In certain embodiments, the anti-TNFα antibody or antigen-binding fragment thereof comprises a light chain variable region of an antibody, wherein the light chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity with the light chain variable region of the following molecules, respectively: Adalimumab, Infliximab, Afelimomab, golimumab, or anti-murine TNFαmIgG2a8c11.

[0493] In certain embodiments, the anti-TNFα antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region of an antibody, wherein the heavy chain variable region and the light chain variable region have at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity with the light chain variable region of the following molecules, respectively: Adalimumab, Infliximab, Afelimomab, golimumab, or anti-murine TNFαmIgG2a 8c11.

[0494] In certain embodiments, the anti-TNFα antibody or antigen-binding fragment thereof comprises a light chain of an antibody, wherein the light chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the light chain of adalimumab, infliximab, afelimomab, golimumab, or anti-murine TNFαmIgG2a 8c11, respectively.

[0495] For example, the anti-TNFα antibody or antigen-binding fragment thereof comprises a heavy chain and a light chain of an antibody, wherein the heavy chain and the light chain have at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity with the light chain of the following molecules, respectively: Adalimumab, Infliximab, Afelimomab, golimumab, or anti-murine TNFαmIgG2a 8c11.

[0496] Sequences of exemplary anti-TNF-α antibodies or antigen-binding fragments thereof are provided in Tables 1-2.

[0497] Table 1: Adalimumab amino acid sequence

[0498]

[0499]

[0500] Table 2: Anti-mouse TNFαmIgG2a 8c11 amino acid sequence

[0501]

[0502] In one embodiment, the antibody, or antigen-binding portion thereof, is an antagonist antibody, or antigen-binding portion thereof, that causes a decrease in CD40 activity or function compared to CD40 activity or function in the absence of the antibody, or antigen-binding portion thereof. In specific embodiments, the antibody, or antigen-binding portion thereof, is substantially free of agonist activity, i.e., the antibody, or antigen-binding portion thereof, does not cause an increase in the amount of CD40 activity or function compared to CD40 activity or function in the absence of the antibody, or antigen-binding portion thereof. In certain embodiments, the anti-CD40 antibody is a polyclonal antibody, a monoclonal antibody, a chimeric antibody, a humanized antibody, a human antibody, or an antigen-binding portion thereof.

[0503] In certain embodiments, the anti-CD40 antibody is iscalimab (CFZ533) (Novartis; as described in U.S. Pat. Nos. 8,828,396 and 9,221,913); the anti-CD40 antibody is lucatumumab (Novartis; as described in U.S. Pat. No. 8,277,810); antibodies 5D12, 3A8, and 3C6, or humanized versions thereof (Novartis; as described in U.S. Pat. No. 5,874,082); antibody 15B8 (Novartis; as described in U.S. Pat. No. 7,445,780); antibody 4D11 (Kyowa Hakko Kirin; as described in U.S. Pat. No. 7,193,064); temeliximab (Bristol Myers Squibb; and others). Squibb; as described in U.S. Pat. No. 6,051,228); antibody PG102 (PanGenetics; as described in U.S. Pat. No. 8,669,352); antibody 2C10 (Primatope; U.S. Patent Application Publication No. 20140093497); anti-CD40 antibodies described in U.S. Pat. Nos. 8,591,900 and 8,778,345 (Boehringer Ingelheim); anti-CD40 antibodies described in U.S. Pat. No. 5,801,227 (Amgen); or APX005 (Boehringer Ingelheim; as described in U.S. Pat. App. Pub. No. 20120301488).

[0504] In certain embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) of an antibody, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, each of which has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the HCDR1, HCDR2, and HCDR3 of the following molecule, respectively: Iscalimab (CFZ533).

[0505] In certain embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises a heavy chain variable region of an antibody, wherein the heavy chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the heavy chain variable region of the following molecule: Iscalimab (CFZ533), respectively.

[0506] In certain embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises a heavy chain of an antibody, wherein the heavy chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity, respectively, to the heavy chain of Iscalimab (CFZ533).

[0507] In certain embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises a light chain variable region (VL) of an antibody, wherein the light chain variable region comprises LCDR1, LCDR2, and LCDR3, each of which has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the LCDR1, LCDR2, and LCDR3 of the following molecules, respectively: Iscalimab (CFZ533).

[0508] In certain embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises a light chain variable region of an antibody, wherein the light chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the light chain variable region of the following molecule: Iscalimab (CFZ533), respectively.

[0509] In certain embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises a light chain of an antibody, wherein the light chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the light chain of Iscalimab (CFZ533), respectively.

[0510] Sequences of exemplary anti-CD40 antibodies or antigen-binding fragments thereof are provided in Table 3.

[0511] Table 3: CFZ533 mAb amino acid sequence (Kabat)

[0512]

[0513] In one embodiment, the antibodies of the present invention are specific for (i.e., specifically bind to) IFNAR1. Such antibodies may also be referred to herein as "anti-IFNAR1 antibodies of the present invention." In another embodiment, the antibodies of the present invention are specific for human IFNAR1. In another embodiment, the anti-IFNAR1 antibodies of the present invention may cross-react with IFNAR1 of species other than human or other proteins structurally related to human IFNAR1 (e.g., human IFNAR1 homologs). In other embodiments, the anti-IFNAR1 antibodies of the present invention may only be specific for human IFNAR1 and may not exhibit species or other types of cross-reactivity.

[0514] Selected sequences for anti-IFNAR1 antibodies can be found in US Patent No. 5,235,038, US Patent Application No. 5,919,453. The entire contents of US Patent Nos. 10831,459, 10 / 182,058, 11157,494 and 11521,102 are incorporated herein by reference in their entirety for all purposes.

[0515] In certain embodiments, the anti-IFNAR1 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) of an antibody, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, each of which has at least about 80% sequence identity with the HCDR1, HCDR2, and HCDR3 of the following molecule, respectively: Anifrolumab (MEDI-546).

[0516] In certain embodiments, the anti-IFNAR1 antibody or antigen-binding fragment thereof comprises a heavy chain variable region of an antibody, wherein the heavy chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity with the heavy chain variable region of the following molecules, respectively: Anifrolumab (MEDI-546).

[0517] In certain embodiments, the anti-IFNAR1 antibody or antigen-binding fragment thereof comprises a heavy chain of an antibody, wherein the heavy chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity, respectively, to the heavy chain of Anifrolumab (MEDI-546).

[0518] In certain embodiments, the anti-IFNAR1 antibody or antigen-binding fragment thereof comprises a light chain variable region (VL) of an antibody, wherein the light chain variable region comprises LCDR1, LCDR2, and LCDR3, each of which has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity with the LCDR1, LCDR2, and LCDR3 of the following molecules, respectively: Anifrolumab (MEDI-546).

[0519] In certain embodiments, the anti-IFNAR1 antibody or antigen-binding fragment thereof comprises a light chain variable region of an antibody, wherein the light chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the light chain variable region of the following molecules, respectively: Anifrolumab (MEDI-546).

[0520] In certain embodiments, the anti-IFNAR1 antibody or antigen-binding fragment thereof comprises a light chain of an antibody, wherein the light chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the light chain of the following molecule, respectively: Anifrolumab (MEDI-546).

[0521] Sequences of exemplary anti-IFNAR1 antibodies or antigen-binding fragments thereof are provided in Table 4.

[0522] Table 4: Anifrolumab monoclonal antibody amino acid sequence (Kabat)

[0523]

[0524]

[0525] In certain embodiments, the anti-BDCA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region of an antibody, wherein the heavy chain variable region has at least about 80% sequence identity with the heavy chain variable region of the following molecule: litifilimab (BIIB059). In certain embodiments, the anti-BDCA2 antibody or antigen-binding fragment thereof comprises a light chain variable region of an antibody, wherein the light chain variable region has at least about 80% sequence identity with the light chain variable region of the following molecule: litifilimab (BIIB059).

[0526] In certain embodiments, the anti-BDCA2 antibody or antigen-binding fragment thereof comprises a heavy chain of an antibody, wherein the heavy chain has at least about 80% sequence identity with the heavy chain of the following molecule: Litifilimab (BIIB059). In certain embodiments, the anti-BDCA2 antibody or antigen-binding fragment thereof comprises a light chain of an antibody, wherein the light chain has at least about 80% sequence identity with the light chain of the following molecule: Litifilimab (BIIB059).

[0527] In certain embodiments, the anti-BDCA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) of an antibody, wherein the heavy chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the heavy chain variable region of Litifilimab (BIIB059), respectively.

[0528] In certain embodiments, the anti-BDCA2 antibody or antigen-binding fragment thereof comprises a light chain variable region (VL) of an antibody, wherein the light chain variable region has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the light chain variable region of Litifilimab (BIIB059), respectively.

[0529] In certain embodiments, the anti-BDCA2 antibody or antigen-binding fragment thereof comprises a heavy chain of an antibody, wherein the heavy chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the heavy chain of Litifilimab (BIIB059), respectively.

[0530] In certain embodiments, the anti-BDCA2 antibody or antigen-binding fragment thereof comprises a light chain of an antibody, wherein the light chain has at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to the light chain of Litifilimab (BIIB059), respectively.

[0531] Sequences of exemplary anti-BDCA2 antibodies or antigen-binding fragments thereof are provided in Table 5.

[0532] Table 5: Litifilimab (BIIB059) mAb amino acid sequence

[0533]

[0534]

[0535] The present application also encompasses variants and equivalents that are substantially homologous to the anti-TNF-α antibodies, anti-CD40 antibodies, or anti-IFNAR1 antibodies described herein. These variants and equivalents may contain, for example, conservative substitution mutations, i.e., one or more amino acids are replaced by similar amino acids. For example, a conservative substitution refers to an amino acid being replaced by another amino acid within the same general category, such as an acidic amino acid being replaced by another acidic amino acid, a basic amino acid being replaced by another basic amino acid, or a neutral amino acid being replaced by another neutral amino acid. The purpose of conservative amino acid substitutions is well known in the art. The antibody may be a recombinant polypeptide, a natural polypeptide, or a synthetic polypeptide of the antibody. It will be understood in the art that some amino acid sequences of the present application may be changed without significantly affecting the structure or function of the protein. Therefore, the present application further includes variations of the polypeptide that exhibit significant activity or that include regions of the antibody. Such mutants include deletions, insertions, inversions, duplications, and type substitutions.

[0536] In certain embodiments, the ligand is selected from: Adalimumab, Iscalimab (CFZ533), Anifrolumab (MEDI-546), Infliximab, Afelimomab, Golimumab, Anti-mouse TNFαmIgG2a 8c11, Litifilimab (BIIB059), derivatives thereof, and biosimilars thereof.

[0537] For example, the ligand is selected from the group consisting of: Adalimumab, Iscalimab (CFZ533) and Anifrolumab (MEDI-546).

[0538] In certain embodiments, the ligand-drug conjugate has the following structure:

[0539]

[0540] in,

[0541] Ab represents a ligand capable of binding to a target, including but not limited to antibodies and antigen-binding fragments thereof;

[0542] N a-I Any number from 1 to 10;

[0543] Tr does not exist or Tr is any group;

[0544] L3 is selected from a polypeptide fragment;

[0545] L2 is absent or selected from a connecting fragment;

[0546] L1 is selected from a coupling unit;

[0547] R1, R2, R3, R4 and R5 are each independently any group;

[0548] B does not exist or B is any group;

[0549] W does not exist or W is any group;

[0550] Each CR4R5 unit is independently unsubstituted or substituted by a member selected from the group consisting of -O-, -S-, -NR-, -S(O)-, -S(O)2-, -C(=O)-, -C=C-, and -C≡C-, wherein R4 and R5 may together form an optionally substituted cycloalkyl, an optionally substituted heterocyclyl, an optionally substituted bridged ring group, an optionally substituted heterobridged ring group, an optionally substituted spirocyclyl, and an optionally substituted heterospirocyclyl, and n is any integer from 1 to 20;

[0551] X is selected from: -O-, -S-, -NR-;

[0552] Y1 is any group, m is any integer from 0 to 4;

[0553] Y2 is selected from -O-, -S- and -NR-;

[0554] wherein the substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O)OR, -S(O)2NHR, -S( O)2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy; wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0555] The term "ligand" generally means any molecule that is capable of specifically binding to and or reactively binding to or complexing with a target molecule, such as a receptor, substrate, antigenic determinant, or other binding site on a target cell or tissue. Examples of ligands include antibodies and fragments thereof (e.g., monoclonal antibodies or fragments thereof), enzymes (e.g., fibrinolytic enzymes), biological response modifiers (e.g., interleukins, interferons, erythropoietins, or colony stimulating factors), peptide hormones and antigen-binding fragments thereof, polysaccharides, lipids, oligonucleotides, polynucleotides, synthetic molecules, inorganic molecules, organic molecules, and any combination thereof.

[0556] The term "target" or "target molecule" can include a wide variety of substances and molecules ranging from simple molecules to complex targets. A target molecule can be a protein, nucleic acid, lipid, carbohydrate, or any other molecule that can be recognized by a polypeptide domain. For example, a target molecule can include a compound (i.e., a non-biological compound, e.g., an organic molecule, an inorganic molecule, or a molecule with organic and inorganic atoms, but not including polynucleotides and proteins), a mixture of a compound, an array of spatially located compounds, a biomacromolecule, a phage peptide display library, a polyribosomal peptide display library, an extract made from a biological material (e.g., bacteria, plants, fungi, or animal (e.g., mammalian) cells or tissues), a protein, a toxin, a peptide hormone, a cell, a virus, etc. Other target molecules include, for example, whole cells, whole tissues, a mixture of related or unrelated proteins, a mixture of viruses or bacterial strains, etc.

[0557] The term "specific binding" or "specific" generally refers to a measurable and reproducible interaction, such as binding between a target and an antibody, that can determine the presence of a target in the presence of a heterogeneous population of molecules (including biomolecules). For example, an antibody that specifically binds to a target (which may be an epitope) may be one that binds to that target with greater affinity, avidity, more readily, and / or for a greater duration than it binds to other targets. In certain embodiments, an antibody specifically binds to an epitope on a protein that is conserved among proteins of different species. In certain embodiments, specific binding may include, but does not require, exclusive binding.

[0558] The structure of the coupling unit L1 may change before and after coupling with the ligand, that is, in the linker-payload structure (such as Formula IVa or Formula IVb) and the drug-ligand conjugate structure (such as Formula Va or Formula Vb), the structure of L1 may change, and such changes can be easily determined by a person skilled in the art.

[0559] For example, when L1 is coupled to the ligand via a sulfhydryl group, the structure of L1 changes as follows:

[0560]

[0561]

[0562] Wherein, the thiol group can be derived from a ligand, and the R L1a 、R L1b 、R L1c Each is independently selected from: hydrogen, optionally substituted alkyl and optionally substituted aryl; for example, the R L1a 、R L1b 、R L1c and optionally substituted benzyl.

[0563] For example, when L1 is coupled to the ligand via the amino group, the structure of L1 changes as follows:

[0564]

[0565]

[0566] The amino group may be derived from a ligand.

[0567] For example, when L1 is coupled to the ligand via click chemistry, the structure of L1 changes as follows:

[0568]

[0569]

[0570] For example, wherein R1 and R2 can be independently selected from: H, F, Cl, Br and optionally substituted methyl.

[0571] For example, wherein said R3 can be selected from: optionally substituted -CH2Cl, optionally substituted -CH2SH, optionally substituted -CH2OH, optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted optionally substituted -OH, optionally substituted -OCH3, optionally substituted -OCH2F, optionally substituted -OCH2Cl, optionally substituted -OCH2CN, optionally substituted -OCH2CH3, optionally substituted thiol, optionally substituted -SCH2F, optionally substituted -SCH2Cl, optionally substituted -SCH2CF3, and optionally substituted -SCH2CN.

[0572] For example, wherein said B can be selected from:

[0573] Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted Optionally substituted wherein the substituents are selected from the group consisting of: protium, deuterium, tritium, halogen, -CN, =O, =N-OH, =N-OR, =NR, -OR, -C(O)R, -C(O)OR, -OC(O)R, -OC(O)OR, -C(O)NHR, -C(O)NR2, -OC(O)NHR, -OC(O)NR2, -SR, -S(O)R, -S(O)2R, -NHR, -N(R)2, -NHC(O)R, -NRC(O)R, -NHC(O)OR, -NRC(O)OR, -S(O)2NHR, -S (O)2N(R)2, -NHS(O)2NR2, -NRS(O)2NR2, -NHS(O)2R, -NRS(O)2R, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy, wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0574] For example, the W may be absent or W may be selected from:

[0575]

[0576] For example, the conjugate can be selected from the following structures:

[0577]

[0578]

[0579]

[0580]

[0581]

[0582]

[0583]

[0584]

[0585]

[0586]

[0587]

[0588]

[0589]

[0590]

[0591]

[0592]

[0593]

[0594]

[0595]

[0596]

[0597]

[0598]

[0599]

[0600]

[0601]

[0602]

[0603]

[0604]

[0605]

[0606]

[0607]

[0608]

[0609]

[0610]

[0611]

[0612]

[0613]

[0614]

[0615]

[0616]

[0617]

[0618]

[0619]

[0620]

[0621]

[0622]

[0623]

[0624]

[0625]

[0626]

[0627]

[0628]

[0629]

[0630]

[0631]

[0632]

[0633]

[0634]

[0635]

[0636] ; where N a-I Any number from 1 to 10.

[0637] In certain embodiments, the conjugate of the present invention, wherein the ligand-drug conjugate has the following structure:

[0638]

[0639] in,

[0640] Ab represents a ligand capable of binding to a target, preferably an antibody or an antigen-binding fragment thereof;

[0641] N a-I Any number from 1 to 10;

[0642] Said X is selected from -O-, -S- and -NH-;

[0643] R4 and R5 are each independently selected from the group consisting of H, F, Cl, -OH, -NH2, and C1-C6 alkyl;

[0644] Said n is selected from 1, 2 or 3;

[0645] Said Y1 is absent or selected from: hydroxyl, thiol, amino and C1-C6 alkyl;

[0646] Said R1 and R2 are each independently selected from hydrogen, fluorine, chlorine and methyl;

[0647] The R3 is selected from: -CH2Cl, -CH2SH, -CH2OH, -OCH3, -OCH2F, -OCH2Cl, -OCH2CN, -OCH2CH3, -SH, -SCH2F, -SCH2Cl, -SCH2CF3, and -SCH2CN.

[0648] In certain embodiments, the conjugate of the present invention is selected from the following structures:

[0649]

[0650]

[0651] ; where N a-I is any number from 1 to 10, and Ab is selected from an antibody or an antigen-binding fragment thereof.

[0652] In certain embodiments, the conjugate of the present invention is selected from the following structures:

[0653]

[0654]

[0655]

[0656] where N a-I Any number from 1 to 10.

[0657] In certain embodiments, the active metabolite of the conjugate comprises the aforementioned compound.

[0658] On the other hand, the present application provides a pharmaceutical composition comprising the aforementioned compound or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof and / or the aforementioned conjugate, and optionally a pharmaceutically acceptable carrier.

[0659] On the other hand, the present application provides a method for influencing immune system function, comprising administering to a subject the aforementioned compound or its tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, the aforementioned conjugate and / or the aforementioned pharmaceutical composition.

[0660] In certain embodiments, affecting immune system function comprises affecting immune cell function.

[0661] In certain embodiments, the immune cells are selected from the group consisting of granulocytes and agranulocytes.

[0662] In certain embodiments, the immune cell is selected from the group consisting of neutrophils, eosinophils, and basophils.

[0663] In certain embodiments, the immune cell is selected from the group consisting of a lymphocyte and a phagocyte.

[0664] In certain embodiments, the immune cell is selected from the group consisting of a B cell, a T cell, a natural killer cell, a monocyte, a macrophage, a mast cell, and a dendritic cell.

[0665] On the other hand, the present application provides the use of the aforementioned compound or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, the aforementioned conjugate and / or the aforementioned pharmaceutical composition in the preparation of a medicament for preventing and / or treating a disease and / or symptom.

[0666] In certain embodiments, the disease and / or condition comprises a disease and / or condition associated with glucocorticoid receptor signaling.

[0667] In certain embodiments, the disease and / or condition is selected from the group consisting of a proliferative disease and / or condition, a metabolic disease and / or condition, an inflammatory disease and / or condition, and a neurodegenerative disease and / or condition.

[0668] In certain embodiments, the disease and / or symptom is selected from the following groups: systemic autoimmune diseases and / or symptoms, blood system-related diseases and / or symptoms, neuromuscular system-related diseases and / or symptoms, digestive system-related diseases and / or symptoms, urinary system-related diseases and / or symptoms, endocrine gland system-related diseases and / or symptoms, skin and muscle system-related diseases and / or symptoms, and respiratory system-related diseases and / or symptoms.

[0669] In certain embodiments, the disease and / or condition is selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosus, scleroderma, Sjögren's syndrome, ankylosing spondylitis, Wegener's granulomatosis, and systemic sclerosis.

[0670] In certain embodiments, the disease and / or condition is selected from the group consisting of autoimmune hemolytic anemia, pernicious anemia, idiopathic thrombocytopenic purpura, idiopathic thrombocytopenia, and vasculitis.

[0671] In certain embodiments, the disease and / or condition is selected from the group consisting of multiple sclerosis, myasthenia gravis, and Guillain-Barré syndrome.

[0672] In certain embodiments, the disease and / or condition is selected from the group consisting of ulcerative colitis, Crohn's disease, autoimmune liver disease, and atrophic gastritis.

[0673] In certain embodiments, the disease and / or condition is selected from the group consisting of IgA nephropathy, primary nephrotic syndrome, autoimmune glomerulonephritis, good-of-bowel syndrome, and lupus nephritis.

[0674] In certain embodiments, the disease and / or condition is selected from the group consisting of type I diabetes, Grave's disease, Hashimoto's thyroiditis, primary adrenal atrophy, and chronic thyroiditis.

[0675] In certain embodiments, the disease and / or condition is selected from the group consisting of psoriasis, herpes vulgaris, cutaneous lupus erythematosus, dermatomyositis, and polymyalgia rheumatica.

[0676] In certain embodiments, the disease and / or condition is asthma.

[0677] On the other hand, the present application provides a method for preventing and / or treating a disease and / or symptom, comprising administering to a subject in need thereof the aforementioned compound or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, the aforementioned conjugate and / or the aforementioned pharmaceutical composition.

[0678] In certain embodiments, the disease and / or condition comprises a disease and / or condition associated with glucocorticoid receptor signaling.

[0679] In certain embodiments, the disease and / or condition is selected from the group consisting of a proliferative disease and / or condition, a metabolic disease and / or condition, an inflammatory disease and / or condition, and a neurodegenerative disease and / or condition.

[0680] In certain embodiments, the disease and / or symptom is selected from the following groups: systemic autoimmune diseases and / or symptoms, blood system-related diseases and / or symptoms, neuromuscular system-related diseases and / or symptoms, digestive system-related diseases and / or symptoms, urinary system-related diseases and / or symptoms, endocrine gland system-related diseases and / or symptoms, skin and muscle system-related diseases and / or symptoms, and respiratory system-related diseases and / or symptoms.

[0681] In certain embodiments, the disease and / or condition is selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosus, scleroderma, Sjögren's syndrome, ankylosing spondylitis, Wegener's granulomatosis, and systemic sclerosis.

[0682] In certain embodiments, the disease and / or condition is selected from the group consisting of autoimmune hemolytic anemia, pernicious anemia, idiopathic thrombocytopenic purpura, idiopathic thrombocytopenia, and vasculitis.

[0683] In certain embodiments, the disease and / or condition is selected from the group consisting of multiple sclerosis, myasthenia gravis, and Guillain-Barré syndrome.

[0684] In certain embodiments, the disease and / or condition is selected from the group consisting of ulcerative colitis, Crohn's disease, autoimmune liver disease, and atrophic gastritis.

[0685] In certain embodiments, the disease and / or condition is selected from the group consisting of IgA nephropathy, primary nephrotic syndrome, autoimmune glomerulonephritis, good-of-bowel syndrome, and lupus nephritis.

[0686] In certain embodiments, the disease and / or condition is selected from the group consisting of type I diabetes, Grave's disease, Hashimoto's thyroiditis, primary adrenal atrophy, and chronic thyroiditis.

[0687] In certain embodiments, the disease and / or condition is selected from the group consisting of psoriasis, herpes vulgaris, cutaneous lupus erythematosus, dermatomyositis, and polymyalgia rheumatica.

[0688] In certain embodiments, the disease and / or symptom is asthma. Without being bound by any theory, the following examples are merely for illustrating the compounds, preparation methods and uses of the present application, and are not intended to limit the scope of the present invention.

[0689] Example

[0690] In this application,

[0691] Reagents providing bromination conditions include, but are not limited to, bromine water, N-bromosuccinimide, dibromohydantoin, phosphorus tribromide, liquid bromine, liquid bromine / triphenylphosphine, hydrobromic acid, and carbon tetrabromide;

[0692] Titanium catalysts include, but are not limited to, tetraisopropyl titanate, triisopropoxytitanium chloride, titanium tetrachloride, and methyl titanium triisopropoxide;

[0693] Palladium catalysts include, but are not limited to, tetrakistriphenylphosphine palladium, palladium acetate, palladium chloride, bis(triphenylphosphine)palladium dichloride, tris(dibenzylideneacetone)dipalladium, bis(dibenzylideneacetone)dipalladium, bis(acetonitrile)palladium dichloride, [1,1'-bis{diphenylphosphino}ferrocene]palladium dichloride, [1,1'-bis{diphenylphosphino}ferrocene]palladium dichloride dichloromethane complex, dibenzonitrilepalladium dichloride, 1,4-bis(diphenylphosphino)butane-palladium chloride, allylpalladium chloride dimer, and allylcyclopentadienylpalladium;

[0694] Boronate dimers include, but are not limited to, bis(2-methyl-2,4-pentanediol) borate, bis(diisopropyl-L-tartrate diethyl) borate, bis[(-)pinanediol]diboronate, bis(1S,2S,3R,5S)(+)-pinanediol diboronate, tetramethylaminodiborane, bis(N,N,N',N'-tetramethyl-D-tartrate)amide diboron, tetrahydroxydiboron, bis(N,N,N',N'-tetramethyl-L-tartrate diolate)diboron, bis(diisopropyl-D-tartaric acid glycolate)diboron, bis(D-tartrate diethyl)borate, bis(2,4-dimethyl-2,4-pentanediol)borate, bis(L-tartrate diethyl)borate, 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-ylboronic acid;

[0695] Metal copper salts include, but are not limited to, copper sulfate, copper sulfate pentahydrate, cuprous sulfate, cupric chloride, cuprous chloride, copper carbonate, copper phosphate, copper acetate and its hydrates, copper oxalate, copper fluoroborate and its hydrates, copper dimethoxide, copper tartrate, copper formate, cuprous iodide, copper trifluoroacetate, copper trifluoromethanesulfonate, basic copper carbonate, copper bromide, cuprous bromide, and cuprous oxide;

[0696] The ligand can be selected from any ligand commonly used in the Ullmann reaction, including but not limited to L-proline, tyrosine, phenylalanine, 1,10-phenanthroline, N,N'-dimethylethylenediamine, ethylene glycol, 1,1'-binaphthyl-2,2'-diol, ethyl 2-carbonylcyclohexylcarboxylate, and salicylaldehyde hydrazone;

[0697] The condensing agent can be selected from 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride, 1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N,N'-dicyclohexylcarbodiimide, N,N'-diisopropylcarbodiimide, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazole, O-benzotriazole-N,N, N',N'-tetramethyluronium hexafluorophosphate, 2-(7-azobenzotriazole))-N,N,N',N'-tetramethyluronium hexafluorophosphate, benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate or benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate, preferably 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride or 1-hydroxybenzotriazole and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride;

[0698] Reagents providing alkaline conditions include organic bases and inorganic bases, the organic bases include but are not limited to triethylamine, diethylamine, N-methylmorpholine, pyridine, hexahydropyridine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, potassium acetate, sodium tert-butoxide, potassium tert-butoxide, etc.; the inorganic bases include but are not limited to sodium hydride, potassium carbonate, sodium carbonate, cesium carbonate, sodium hydroxide, lithium hydroxide, sodium phosphate, potassium phosphate;

[0699] Reagents providing acidic conditions include protonic acids and Lewis acids, the protonic acids include but are not limited to hydrochloric acid, sulfuric acid, nitric acid, nitrous acid, sulfurous acid, phosphoric acid, phosphorous acid, formic acid, acetic acid, propionic acid, butyric acid, citric acid, benzoic acid, p-toluenesulfonic acid, p-nitrobenzoic acid, methanesulfonic acid, trifluoromethanesulfonic acid, trifluoroacetic acid; the Lewis acids include but are not limited to boron trifluoride, zinc chloride, magnesium chloride, aluminum chloride, tin chloride, ferric chloride;

[0700] Hydrogenation conditions include, but are not limited to: Pb / C / hydrogen, Pt / C / hydrogen, palladium chloride / hydrogen, Raney nickel / hydrogen, palladium hydroxide on carbon / hydrogen, palladium hydroxide / hydrogen;

[0701] Reagents providing oxidation conditions include, but are not limited to, Dess-Martin periodinane, hydrogen peroxide, sodium chlorite, sodium hypochlorite, and potassium perchlorate;

[0702] Reagents providing reducing conditions include, but are not limited to, sodium hydride, calcium hydride, lithium hydride, lithium aluminum hydride, sodium borohydride, lithium borohydride, sodium triethylborohydride, sodium triacetoxyborohydride, and sodium cyanoborohydride;

[0703] Reagents providing oxidative conditions include, but are not limited to, Dess-Martin periodinane, hydrogen peroxide, sodium chlorite, sodium hypochlorite, and potassium perchlorate.

[0704] In addition, in this application

[0705] The structures of the compounds were determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR measurements were performed using a Quantum-I NMR spectrometer using deuterated dimethyl sulfoxide (DMSO-D), deuterated chloroform (CDC13), or deuterated methanol (CD3OD) as the solvents, with tetramethylsilane (TMS) as the internal standard. Chemical shifts are given in units of 10_6 (ppm).

[0706] MS was measured using an Angilent 6230 ESI-TOF mass spectrometer (manufacturer: Agilent, model: 6230).

[0707] UPLC analysis was performed using a Waters Acquity UPLC SQD liquid chromatography-mass spectrometer (Poroshell 120 EC-C18, 2.1 mm x 50 mm, 1.9 μm column).

[0708] HPLC analysis was performed using an Agilent 1260 high pressure liquid chromatograph (TOSOH G3000 SW SEC column).

[0709] UV was measured using a Thermonanodrop 2000 UV spectrophotometer.

[0710] Enzyme-linked immunosorbent assay was performed using EnVision microplate reader (PerkinElmer).

[0711] The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The specifications of the silica gel plate used in thin layer chromatography (TLC) are 0.15mm0.2mm, and the specifications used for thin layer chromatography separation and purification products are 0.4mm0.5mm silica gel plates.

[0712] Column chromatography generally uses Yantai Huanghai 200-300 mesh silica gel as the carrier.

[0713] The known starting materials of the present application can be synthesized by methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organnics, Aldrich Chemical Company, Accela Chem Bio Inc, Darui Chemicals and other companies.

[0714] Unless otherwise specified in the examples, all reactions were carried out under an argon or nitrogen atmosphere.

[0715] Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a capacity of about 1 L.

[0716] Hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon with a capacity of about 1L.

[0717] Unless otherwise specified in the examples, the solution in the reaction refers to an aqueous solution.

[0718] Unless otherwise specified in the examples, the reaction temperature is room temperature, which is the most suitable reaction temperature and ranges from 20°C to 30°C.

[0719] The eluent system for column chromatography and the developing solvent system for thin-layer chromatography used for purifying compounds include: A: dichloromethane and isopropanol system, B: dichloromethane and methanol system, and C: petroleum ether and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of triethylamine and acidic or alkaline reagents can also be added for adjustment.

[0720] Some compounds of the present disclosure were characterized by TOF-LC / MS using an Agilent 6230 time-of-flight mass spectrometer and an Agilent 1290-Infinity ultra-performance liquid chromatograph.

[0721] The exemplary preparation route of the present application is as follows:

[0722] 1. Preparation of payload and linker-payload

[0723] Preparation Route 1

[0724]

[0725] Step 1: introducing a bromine atom onto the benzene ring of the compound of formula (P1) under optional bromination conditions;

[0726] Step 2: The compound of formula (P2) reacts with a borate dimer under optional palladium reagent catalysis to obtain a compound of formula (P3);

[0727] Step 3: reacting the compound of formula (P3) with the compound of formula (Y1) under optional palladium reagent catalysis conditions to obtain a compound of formula (P4);

[0728] Step 4: reacting the compound of formula (P4) with the compound of formula (Y2) under optional acidic or alkaline conditions to obtain a compound of formula (P5);

[0729] Step 5: The compound of formula (P5) is subjected to acidic or alkaline conditions to remove the protective group PG, and separated by preparative HPLC to obtain the compound of formula (P6).

[0730] in:

[0731] PG can be a protecting group for common hydroxyl groups;

[0732] Y is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -OR, -SR, -NHR, -N(R)2, -PHR, -P(R)2, -P(=O)HR, -P(=O)R2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl;

[0733] Ring B is an optionally substituted aryl or heteroaryl group;

[0734] -B(OR)2 is a borate-based monomer, wherein two Rs may be linked to form a heterocyclic ring, a heterobridged ring, or a heterospirocyclic ring, and the ring may be optionally substituted with a C1-C6 alkyl group, an aryl group, a heteroaryl group, a carboxyl group, or an acyloxy C1-C6 alkyl group;

[0735] X is chlorine, bromine or iodine;

[0736] wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy;

[0737] R, R1, R2 and R3 can be respectively defined as R, R1, R2 and R3 shown in any one of Formula (I), Formula (IIa), Formula (IIb), Formula (IIIa), Formula (IIIb), Formula (IVa) or Formula (IVb) of the present application.

[0738] Preparation Route 2

[0739]

[0740] Step 1: reacting a compound of formula (Y1) with a compound (P1) under optional alkaline conditions with the catalysis of an optional metal copper salt and an optional ligand to obtain a compound of formula (P2);

[0741] Step 2: reacting the compound of formula (P2) under optional reducing conditions to obtain the compound of formula (P3);

[0742] Step 3: The compound of formula (P3) is oxidized under optional conditions to obtain the compound of formula (P4);

[0743] Step 4: reacting the compound of formula (P4) with the compound of formula (Y2) under optional acidic or alkaline conditions to obtain the compound of formula (P5);

[0744] Step 5: The compound of formula (P5) is subjected to acidic or alkaline conditions to remove the protective group PG, and separated by preparative HPLC to obtain the compound of formula (P6).

[0745] in:

[0746] PG can be a protecting group for common hydroxyl groups;

[0747] Y is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -OR, -SR, -NHR, -N(R)2, -PHR, -P(R)2, -P(=O)HR, -P(=O)R2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl;

[0748] Ring B is an optionally substituted aryl or heteroaryl group;

[0749] R1, R2 and R3 can be defined as R1, R2 and R3 shown in any one of Formula (I), Formula (IIa), Formula (IIb), Formula (IIIa), Formula (IIIb), Formula (IVa) or Formula (IVb) of the present application, respectively.

[0750] Preparation Route 3

[0751]

[0752] The first step: the compound of formula (P1) undergoes carbonyl insertion reaction under optional palladium reagent catalysis conditions to obtain a compound of formula (P2);

[0753] Step 2: reacting the compound of formula (P2) under optional reducing conditions to obtain the compound of formula (P3);

[0754] Step 3: The compound of formula (P3) is optionally brominated to obtain the compound of formula (P4);

[0755] Step 4: reacting the compound of formula (P4) with the compound of formula (Y1) under optional alkaline conditions to obtain a compound of formula (P5);

[0756] Step 5: reacting the compound of formula (P5) under optional reducing conditions to obtain the compound of formula (P6);

[0757] Step 6: The compound of formula (P6) is subjected to optional oxidation conditions to obtain the compound of formula (P7);

[0758] Step 7: reacting the compound of formula (P7) with the compound of formula (Y2) under optional acidic or alkaline conditions to obtain the compound of formula (P8);

[0759] Step 8: The compound of formula (P8) is subjected to acidic or alkaline conditions to remove the protective group PG, and separated by preparative HPLC to obtain the compound of formula (P9).

[0760] in:

[0761] PG1 and PG2 can be common hydroxyl or ester protecting groups;

[0762] Y is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -OR, -SR, -NHR, -N(R)2, -PHR, -P(R)2, -P(=O)HR, -P(=O)R2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxy, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy;

[0763] R may be an optionally substituted C1-C6 alkyl group;

[0764] Ring B is an optionally substituted aryl or heteroaryl group;

[0765] R, R1, R2 and R3 can be respectively defined as R, R1, R2 and R3 shown in any one of Formula (I), Formula (IIa), Formula (IIb), Formula (IIIa), Formula (IIIb), Formula (IVa) or Formula (IVb) of the present application.

[0766] Preparation Route 4

[0767]

[0768] Step 1: introducing a bromine atom at the methyl group of the compound of formula (P1) under optional bromination conditions to obtain a compound of formula (P2);

[0769] Step 2: The compound of formula (P2) reacts with the compound of formula (Y1) under optional palladium reagent catalysis conditions to obtain a compound of formula (P3);

[0770] Step 3: reacting the compound of formula (P3) with the compound of formula (Y2) under optional acidic or alkaline conditions to obtain a compound of formula (P4);

[0771] Step 4: The compound of formula (P4) is subjected to acidic or alkaline conditions to remove the protective group PG, and separated by preparative HPLC to obtain the compound of formula (P5).

[0772] in:

[0773] PG1 can be a common hydroxyl protecting group;

[0774] Y is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -OR, -SR, -NHR, -N(R)2, -PHR, -P(R)2, -P(=O)HR, -P(=O)R2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0775] Ring B is an optionally substituted aryl or heteroaryl group;

[0776] R1, R2 and R3 can be defined as R1, R2 and R3 shown in any one of Formula (I), Formula (IIa), Formula (IIb), Formula (IIIa), Formula (IIIb), Formula (IVa) or Formula (IVb) of the present application, respectively;

[0777] Preparation Route 5

[0778]

[0779] Step 1: reacting the compound of formula (P1) under optional reducing conditions to obtain the compound of formula (P2);

[0780] Step 2: The compound of formula (P2) is treated with a protective group PG under optional acidic or basic conditions to obtain the compound of formula (P3);

[0781] Step 3: reacting the compound of formula (P3) with the compound of formula (Y1) under optional alkaline conditions to obtain a compound of formula (P4);

[0782] Step 4: reacting the compound of formula (P4) under optional reducing conditions to obtain the compound of formula (P5);

[0783] Step 5: The compound of formula (P5) is subjected to optional oxidation conditions to obtain the compound of formula (P6);

[0784] Step 6: reacting the compound of formula (P6) with the compound of formula (Y2) under optional acidic or alkaline conditions to obtain the compound of formula (P7);

[0785] Step 7: The compound of formula (P7) is subjected to acidic or alkaline conditions to remove the protective group PG, and separated by preparative HPLC to obtain the compound of formula (P8).

[0786] in:

[0787] Y is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -OR, -SR, -NHR, -N(R)2, -PHR, -P(R)2, -P(=O)HR, -P(=O)R2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0788] Ring B is an optionally substituted aryl or heteroaryl group;

[0789] PG1 and PG2 can be common hydroxyl groups or protective groups of hydroxyl groups;

[0790] R1, R2 and R3 can be defined as R1, R2 and R3 shown in any one of Formula (I), Formula (IIa), Formula (IIb), Formula (IIIa), Formula (IIIb), Formula (IVa) or Formula (IVb) of the present application, respectively;

[0791] Preparation Route 6

[0792]

[0793] Step 1: The compound of formula (P1) reacts with the compound of formula (Y1) under optional condensation conditions to obtain the compound of formula (P2)

[0794] Step 2: reacting the compound of formula (P2) under optional reducing conditions to obtain a compound of formula (P3);

[0795] Step 3: reacting the compound of formula (P3) under optional oxidation conditions to obtain a compound of formula (P4);

[0796] Step 4: reacting the compound of formula (P4) with the compound of formula (Y2) under optional acidic or alkaline conditions to obtain the compound of formula (P5);

[0797] Step 5: reacting the compound of formula (P5) under optional reducing conditions to obtain a compound of formula (P6);

[0798] in:

[0799] PG1 and PG2 can be common hydroxyl or ester protecting groups;

[0800] Y is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -OR, -SR, -NHR, -N(R)2, -PHR, -P(R)2, -P(=O)HR, -P(=O)R2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0801] Ring B is an optionally substituted aryl or heteroaryl group;

[0802] R1, R2 and R3 can be defined as R1, R2 and R3 shown in any one of Formula (I), Formula (IIa), Formula (IIb), Formula (IIIa), Formula (IIIb), Formula (IVa) or Formula (IVb) of the present application, respectively;

[0803] Preparation Route 7

[0804]

[0805] Step 1: introducing a nitro group onto the benzene ring of the compound of formula (P1) under nitration conditions to obtain a compound of formula (P2);

[0806] Step 2: The compound of formula (P2) reacts with a borate dimer under optional palladium reagent catalysis to obtain a compound of formula (P3);

[0807] Step 3: reacting the compound of formula (P3) with the compound of formula (Y1) under optional palladium reagent catalysis conditions to obtain a compound of formula (P4);

[0808] Step 4: reacting the compound of formula (P4) with the compound of formula (Y2) under optional acidic or alkaline conditions to obtain a compound of formula (P5);

[0809] Step 5: The compound of formula (P5) is subjected to optional hydrogenation conditions to obtain a compound of formula (P6);

[0810] Step 6: The compound of formula (P6) is subjected to acidic or alkaline conditions to remove the protective group PG, and separated by preparative HPLC to obtain the compound of formula (P7).

[0811] in:

[0812] PG can be a protecting group for common hydroxyl groups;

[0813] -B(OR)2 is a borate-based monomer, wherein two Rs may be linked to form a heterocyclic ring, a heterobridged ring, or a heterospirocyclic ring, and the ring may be optionally substituted with a C1-C6 alkyl group, an aryl group, a heteroaryl group, a carboxyl group, or an acyloxy C1-C6 alkyl group;

[0814] wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy;

[0815] Ring B is an optionally substituted aryl or heteroaryl group;

[0816] R1, R2 and R3 can be defined as R1, R2 and R3 shown in any one of Formula (I), Formula (IIa), Formula (IIb), Formula (IIIa), Formula (IIIb), Formula (IVa) or Formula (IVb) of the present application, respectively;

[0817] Preparation Route 8

[0818]

[0819] Step 1: introducing a nitro group onto the benzene ring of the compound of formula (P1) under nitration conditions to obtain a compound of formula (P2);

[0820] Step 2: The compound of formula (P2) reacts with an ethyl Grignard reagent under optional titanium catalysis conditions to obtain a compound of formula (P3);

[0821] Step 3: The compound of formula (P2) reacts with a borate dimer under optional palladium reagent catalysis to obtain a compound of formula (P3);

[0822] Step 4: reacting the compound of formula (P3) with the compound of formula (Y1) under optional palladium reagent catalysis conditions to obtain a compound of formula (P4);

[0823] Step 5: reacting the compound of formula (P4) with the compound of formula (Y2) under optional acidic or alkaline conditions to obtain a compound of formula (P5);

[0824] Step 6: The nitro group of the compound of formula (P5) is reduced under optional hydrogenation conditions, and the mixture is separated by preparative HPLC to obtain the compound of formula (P6).

[0825] in:

[0826] Ring B is an optionally substituted aryl or heteroaryl group;

[0827] -B(OR)2 is a borate-based monomer, wherein two Rs may be linked to form a heterocyclic ring, a heterobridged ring, or a heterospirocyclic ring, and the ring may be optionally substituted with a C1-C6 alkyl group, an aryl group, a heteroaryl group, a carboxyl group, or an acyloxy C1-C6 alkyl group;

[0828] wherein each R is independently selected from hydrogen, protium, deuterium, tritium, oxygen, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy;

[0829] R1, R2 and R3 can be defined as R1, R2 and R3 shown in any one of Formula (I), Formula (IIa), Formula (IIb), Formula (IIIa), Formula (IIIb), Formula (IVa) or Formula (IVb) of the present application, respectively;

[0830] Preparation Route 9

[0831]

[0832] Step 1: reacting a compound of formula (P1) with a compound of formula (Y1) under optionally alkaline conditions to obtain a compound of formula (P2);

[0833] Step 2: reacting the compound of formula (P2) under optional reducing conditions to obtain the compound of formula (P3);

[0834] Step 3: The compound of formula (P3) is oxidized under optional conditions to obtain the compound of formula (P4);

[0835] Step 4: The compound of formula (P4) reacts with the compound of formula (Y2) under optional acidic or alkaline conditions to obtain the compound of formula (P5)

[0836] Step 5: The compound of formula (P5) is subjected to acidic or alkaline conditions to remove the protective group PG, and separated by preparative HPLC to obtain the compound of formula (P6).

[0837] in:

[0838] PG can be a protecting group for common hydroxyl groups;

[0839] Y is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -OR, -SR, -NHR, -N(R)2, -PHR, -P(R)2, -P(=O)HR, -P(=O)R2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0840] Ring B is an optionally substituted aryl or heteroaryl group;

[0841] R1, R2 and R3 can be defined as R1, R2 and R3 shown in any one of Formula (I), Formula (IIa), Formula (IIb), Formula (IIIa), Formula (IIIb), Formula (IVa) or Formula (IVb) of the present application, respectively;

[0842] Preparation Route 10

[0843]

[0844] Step 1: reacting a compound of formula (P1) with a compound of formula (Y1) under optional palladium catalysis to obtain a compound of formula (P2);

[0845] Step 2: reacting the compound of formula (P2) under optional reducing conditions to obtain the compound of formula (P3);

[0846] Step 3: The compound of formula (P3) is oxidized under optional conditions to obtain the compound of formula (P4);

[0847] Step 4: The compound of formula (P4) reacts with the compound of formula (Y2) under optional acidic or alkaline conditions to obtain the compound of formula (P5)

[0848] Step 5: The compound of formula (P5) is subjected to acidic or alkaline conditions to remove the protective group PG, and separated by preparative HPLC to obtain the compound of formula (P6).

[0849] in:

[0850] X is NR or -O-, R is an optionally substituted C1-C6 alkyl group;

[0851] PG can be a protecting group for common hydroxyl groups;

[0852] Y is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -OR, -SR, -NHR, -N(R)2, -PHR, -P(R)2, -P(=O)HR, -P(=O)R2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0853] Ring B is an optionally substituted aryl or heteroaryl group;

[0854] R1, R2 and R3 can be defined as R1, R2 and R3 shown in any one of Formula (I), Formula (IIa), Formula (IIb), Formula (IIIa), Formula (IIIb), Formula (IVa) or Formula (IVb) of the present application, respectively;

[0855] Preparation Route 11

[0856]

[0857] Step 1: The compound of formula (P1) reacts with the compound of formula (Y1) under optional condensation conditions to obtain the compound of formula (P2)

[0858] Step 2: reacting the compound of formula (P2) under optional reducing conditions to obtain a compound of formula (P3);

[0859] Step 3: reacting the compound of formula (P3) under optional oxidation conditions to obtain a compound of formula (P4);

[0860] Step 4: reacting the compound of formula (P4) with the compound of formula (Y2) under optional acidic or alkaline conditions to obtain the compound of formula (P5);

[0861] Step 5: reacting the compound of formula (P5) under optional reducing conditions and separating by preparative HPLC to obtain the compound of formula (P6);

[0862] in:

[0863] PG can be a protecting group for common hydroxyl groups;

[0864] Y is selected from the group consisting of hydrogen, protium, deuterium, tritium, halogen, -OR, -SR, -NHR, -N(R)2, -PHR, -P(R)2, -P(=O)HR, -P(=O)R2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl; wherein each R is independently selected from the group consisting of hydrogen, protium, deuterium, tritium, oxygen, hydroxyl, halogen, C1-C6 alkyl, C1-C6 alkoxy, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, halogen-substituted C1-C6 alkyl, and halogen-substituted C1-C6 alkoxy.

[0865] R1, R2 and R3 can be defined as R1, R2 and R3 shown in any one of Formula (I), Formula (IIa), Formula (IIb), Formula (IIIa), Formula (IIIb), Formula (IVa) or Formula (IVb) of the present application, respectively;

[0866] Preparation Route 12

[0867]

[0868] Step 1: reacting the compound of formula (P1) with the compound of formula (Y1) under optional condensation conditions to obtain a compound of formula (P2);

[0869] Step 2: The compound of formula (P2) is subjected to removal of the protecting group PG1 under optional acidic or alkaline conditions to obtain the compound of formula (P3).

[0870] Step 3: reacting the compound of formula (P3) with the compound of formula (Y2) under optional condensation conditions to obtain a compound of formula (P4);

[0871] Step 4: removing the protecting group PG1 from the compound of formula (P4) under optional acidic or basic conditions to obtain a compound of formula (P5);

[0872] Step 5: reacting the compound of formula (P5) with the compound of formula (Y3) under optional condensation conditions to obtain a compound of formula (P6);

[0873] Step 6: Removing the protective group PG1 from the compound of formula (P6) under optional acidic or basic conditions to obtain a compound of formula (P7);

[0874] Step 7: The compound of formula (P7) is reacted with the compound of formula (Y3) under optional condensation conditions, and separated by preparative HPLC to obtain a compound of formula (P8).

[0875] in:

[0876] Ring B is an optionally substituted aryl or heteroaryl group;

[0877] W does not exist or W is any group;

[0878] X is -CH2-, -CH(CH3)-, -CH2CH2- or

[0879] Y1 is hydrogen, hydroxy, fluorine, chlorine, methyl, hydroxy or methoxy;

[0880] R1, R2 and R3 may be defined as R1, R2 and R3 shown in any one of formula (I), formula (II), formula (III) or formula (IV) of the present application, respectively;

[0881] LG1, LG2, LG3, and LG4 can be hydroxyl groups of common activated esters or carboxyl groups;

[0882] PG1, PG2 and PG3 can be common amino protecting groups;

[0883] R1, R2 and R3 can be defined as R1, R2 and R3 shown in any one of Formula (I), Formula (IIa), Formula (IIb), Formula (IIIa), Formula (IIIb), Formula (IVa) or Formula (IVb) of the present application, respectively;

[0884] Tr does not exist or Tr is any group;

[0885] L3 is selected from a polypeptide fragment;

[0886] L2 is absent or selected from a connecting fragment;

[0887] L1 is selected from coupling units.

[0888] Preparation Route 13

[0889]

[0890] Step 1: The compound of formula (P1) is reacted with the compound of formula (Y1) under optional condensation conditions, and separated by preparative HPLC to obtain the compound of formula (P2).

[0891] in:

[0892] Ring B is an optionally substituted aryl or heteroaryl group;

[0893] W does not exist or W is any group;

[0894] X is -CH2-, -CH(CH3)-, -CH2CH2- or

[0895] Y is -O-, -S- or -NH-;

[0896] R1, R2 and R3 can be defined as R1, R2 and R3 shown in any one of Formula (I), Formula (IIa), Formula (IIb), Formula (IIIa), Formula (IIIb), Formula (IVa) or Formula (IVb) of the present application, respectively;

[0897] LG1 can be a common activated ester or a hydroxyl group of a carboxyl group;

[0898] Tr does not exist or Tr is any group;

[0899] L3 is selected from a polypeptide fragment;

[0900] L2 is absent or selected from a connecting fragment;

[0901] L1 is selected from coupling units.

[0902] Preparation Route 14

[0903]

[0904] Step 1: reacting the compound of formula (P1) with the compound of formula (Y1) under optional condensation conditions to obtain a compound of formula (P2);

[0905] Step 2: The compound of formula (P2) is subjected to removal of the protecting group PG1 under optional acidic or alkaline conditions to obtain the compound of formula (P3).

[0906] Step 3: reacting the compound of formula (P3) with the compound of formula (Y2) under optional condensation conditions to obtain a compound of formula (P4);

[0907] Step 4: removing the protecting group PG1 from the compound of formula (P4) under optional acidic or basic conditions to obtain a compound of formula (P5);

[0908] Step 5: The compound of formula (P5) is reacted with the compound of formula (Y3) under optional condensation conditions and separated by preparative HPLC to obtain a compound of formula (P6).

[0909] in:

[0910] Ring B is an optionally substituted aryl or heteroaryl group;

[0911] W does not exist or W is any group;

[0912] X is -CH2-, -CH(CH3)-, -CH2CH2- or

[0913] Y is -O-, -S- or -NH-;

[0914] LG1, LG2, and LG3 can be hydroxyl groups of common activated esters or carboxyl groups;

[0915] PG1, PG2 and PG3 can be common amino protecting groups;

[0916] R1, R2 and R3 can be defined as R1, R2 and R3 shown in any one of Formula (I), Formula (IIa), Formula (IIb), Formula (IIIa), Formula (IIIb), Formula (IVa) or Formula (IVb) of the present application, respectively;

[0917] Tr does not exist or Tr is any group;

[0918] L3 is selected from a polypeptide fragment;

[0919] L2 is absent or selected from a connecting fragment;

[0920] L1 is selected from coupling units.

[0921] 2. Preparation of Antibody Drug Conjugates (ADCs)

[0922] Preparation Route 15

[0923]

[0924] The ligand Ab and the compound represented by any of the formulas (IVa) or (IVb) of the present application are reacted in an acidic, neutral or alkaline buffer to obtain a compound represented by formula (Va) or (Vb);

[0925] wherein: Ab is a ligand containing at least one free sulfhydryl group (-SH), wherein the free sulfhydryl group can be obtained by reducing the ligand with a reducing agent; the reducing agent includes but is not limited to tris(2-carboxyethyl)phosphine, mercaptoethanol, dithiothreitol, cysteine, reduced glutathione, etc.; in particular, the disulfide bond (-SS-) between the ligand chains can be reduced to form a free sulfhydryl group; wherein the S atom in the compound represented by formula (IC) or formula (II-C) can be derived from the sulfhydryl group of Ab;

[0926] Tr, L1, L2, L3 can be defined as Tr, L1, L2, L3 in the compound represented by any formula (IVa) or formula (IVb) of the present application; Ab, N a-I Ab, N in the compound represented by any formula (Va) or formula (Vb) of the present application can be respectively a-I Defined; L 1x Represents the structure after coupling with thiol-coupled L1.

[0927] The buffer is selected from the following buffers with a pH of 2 to 12, citric acid-sodium citrate buffer, phosphoric acid-sodium phosphate buffer, phosphoric acid-potassium phosphate buffer, sodium dihydrogen phosphate-disodium hydrogen phosphate buffer, potassium dihydrogen phosphate-dipotassium hydrogen phosphate buffer, succinic acid-sodium succinate buffer, acetic acid-sodium acetate buffer, boric acid-borax buffer, boric acid-potassium borate buffer, borax-sodium hydroxide buffer, histidine-hydrochloric acid buffer, glycine-sodium hydroxide buffer, arginine-hydrochloric acid buffer, sodium bicarbonate-sodium carbonate buffer, potassium bicarbonate-potassium carbonate buffer, Tris-hydrochloric acid buffer, ammonia-ammonium chloride buffer, sodium barbital-hydrochloric acid buffer, borax-sodium carbonate buffer, boric acid-potassium chloride buffer, or a combination of two or more of the above buffers.

[0928] Preparation Route 16

[0929]

[0930] In the first step, the ligand Ab and the compound represented by any of the formulas (IVa) or (IVb) of the present application are reacted in an acidic, neutral or alkaline buffer to obtain a compound represented by the formula (Va) or (Vb);

[0931] wherein: Ab is a ligand containing at least one free sulfhydryl group (-SH), wherein the free sulfhydryl group can be obtained by reducing the ligand with a reducing agent; the reducing agent includes but is not limited to tris(2-carboxyethyl)phosphine, mercaptoethanol, dithiothreitol, cysteine, reduced glutathione, etc.; in particular, the disulfide bond (-SS-) between the ligand chains can be reduced to form a free sulfhydryl group; wherein the S atom in the compound represented by formula (Va) or formula (Vb) can be derived from the sulfhydryl group of Ab;

[0932] In the second step, the compound represented by any of the formulas (Va) or (Vb) of the present application is incubated in an alkaline buffer at a selected temperature for a certain period of time to obtain another compound represented by any of the formulas (Va) or (Vb) of the present application;

[0933] Tr, L2, L3 can be defined as Tr, L2, L3 in the compound represented by any formula (IVa) or formula (IVb) of the present application; Ab, N a-I Ab, N in the compound represented by any formula (Va) or formula (Vb) of the present application can be respectively a-I defined.

[0934] The buffer is selected from the following buffers with a pH of 2 to 12, citric acid-sodium citrate buffer, phosphoric acid-sodium phosphate buffer, phosphoric acid-potassium phosphate buffer, sodium dihydrogen phosphate-disodium hydrogen phosphate buffer, potassium dihydrogen phosphate-dipotassium hydrogen phosphate buffer, succinic acid-sodium succinate buffer, acetic acid-sodium acetate buffer, boric acid-borax buffer, boric acid-potassium borate buffer, borax-sodium hydroxide buffer, histidine-hydrochloric acid buffer, glycine-sodium hydroxide buffer, arginine-hydrochloric acid buffer, sodium bicarbonate-sodium carbonate buffer, potassium bicarbonate-potassium carbonate buffer, Tris-hydrochloric acid buffer, ammonia-ammonium chloride buffer, sodium barbital-hydrochloric acid buffer, borax-sodium carbonate buffer, boric acid-potassium chloride buffer, or a combination of two or more of the above buffers; the selected temperature can be 5 to 60° C.; and the incubation time can be 0 to 72 hours.

[0935] Preparation Route 17

[0936]

[0937] The ligand Ab and a compound having a group capable of coupling with two thiol groups represented by any of formula (IVa) or formula (IVb) of the present application are reacted in an acidic, neutral or alkaline buffer to obtain a compound represented by formula (Va) or formula (Vb);

[0938] wherein: Ab is a ligand containing at least two free sulfhydryl groups (-SH), wherein the free sulfhydryl groups can be obtained by reducing the ligand with a reducing agent; the reducing agent includes but is not limited to tris(2-carboxyethyl)phosphine, mercaptoethanol, dithiothreitol, cysteine, reduced glutathione, etc.; in particular, the disulfide bonds (-SS-) between the ligand chains can be reduced to form free sulfhydryl groups; wherein the S atom in the compound represented by formula (Va) or formula (Vb) can be derived from the sulfhydryl groups of Ab;

[0939] Tr, L1, L2, L3 can be defined as Tr, L1, L2, L3 in the compound represented by any formula (IVa) or formula (IVb) of the present application; Ab, N a-I Ab, N in the compound represented by any formula (Va) or formula (Vb) of the present application can be respectively a-I Defined; L 1y Represents the structure after coupling with two thiol groups of L1.

[0940] The compound represented by formula (Va) or formula (Vb) having a group capable of coupling with two thiol groups may contain L selected from the following group: 1y Group: Each of the R L1a , R L1b and R L1cEach is independently selected from the following group: hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, alkyl, alkenyl, alkynyl, alicyclic, heterocyclic, aryl and heteroaryl.

[0941] The buffer is selected from the following buffers with a pH of 2 to 12, citric acid-sodium citrate buffer, phosphoric acid-sodium phosphate buffer, phosphoric acid-potassium phosphate buffer, sodium dihydrogen phosphate-disodium hydrogen phosphate buffer, potassium dihydrogen phosphate-dipotassium hydrogen phosphate buffer, succinic acid-sodium succinate buffer, acetic acid-sodium acetate buffer, boric acid-borax buffer, boric acid-potassium borate buffer, borax-sodium hydroxide buffer, histidine-hydrochloric acid buffer, glycine-sodium hydroxide buffer, arginine-hydrochloric acid buffer, sodium bicarbonate-sodium carbonate buffer, potassium bicarbonate-potassium carbonate buffer, Tris-hydrochloric acid buffer, ammonia-ammonium chloride buffer, sodium barbital-hydrochloric acid buffer, borax-sodium carbonate buffer, boric acid-potassium chloride buffer, or a combination of two or more of the above buffers.

[0942] Preparation Route 18

[0943]

[0944] The ligand Ab and a compound having a group capable of coupling with two thiol groups represented by any of formula (IVa) or formula (IVb) of the present application are reacted in an acidic, neutral or alkaline buffer to obtain a compound represented by formula (Va) or formula (Vb);

[0945] wherein: Ab is a ligand containing at least one free sulfhydryl group (-SH), wherein the free sulfhydryl group can be obtained by reducing the ligand with a reducing agent; the reducing agent includes but is not limited to tris(2-carboxyethyl)phosphine, mercaptoethanol, dithiothreitol, cysteine, reduced glutathione, etc.; in particular, the disulfide bond (-SS-) between the ligand chains can be reduced to form a free sulfhydryl group; wherein the S atom in the compound represented by formula (Va) or formula (Vb) can be derived from the sulfhydryl group of Ab;

[0946] Tr, L1, L2, L3 can be defined as Tr, L1, L2, L3 in the compound represented by any formula (IVa) or formula (IVb) of the present application; Ab, N a-I Ab, N in the compound represented by any formula (Va) or formula (Vb) of the present application can be respectively a-I Defined; L 1x Represents the structure after coupling with thiol-coupled L1.

[0947] The buffer is selected from the following buffers with a pH of 2 to 12, citric acid-sodium citrate buffer, phosphoric acid-sodium phosphate buffer, phosphoric acid-potassium phosphate buffer, sodium dihydrogen phosphate-disodium hydrogen phosphate buffer, potassium dihydrogen phosphate-dipotassium hydrogen phosphate buffer, succinic acid-sodium succinate buffer, acetic acid-sodium acetate buffer, boric acid-borax buffer, boric acid-potassium borate buffer, borax-sodium hydroxide buffer, histidine-hydrochloric acid buffer, glycine-sodium hydroxide buffer, arginine-hydrochloric acid buffer, sodium bicarbonate-sodium carbonate buffer, potassium bicarbonate-potassium carbonate buffer, Tris-hydrochloric acid buffer, ammonia-ammonium chloride buffer, sodium barbital-hydrochloric acid buffer, borax-sodium carbonate buffer, boric acid-potassium chloride buffer, or a combination of two or more of the above buffers.

[0948] Preparation Route 19

[0949]

[0950] In the first step, the ligand Ab and the compound represented by any of the formulas (IVa) or (IVb) of the present application are reacted in an acidic, neutral or alkaline buffer to obtain a compound represented by the formula (Va) or (Vb);

[0951] wherein: Ab is a ligand containing at least one free sulfhydryl group (-SH), wherein the free sulfhydryl group can be obtained by reducing the ligand with a reducing agent; the reducing agent includes but is not limited to tris(2-carboxyethyl)phosphine, mercaptoethanol, dithiothreitol, cysteine, reduced glutathione, etc.; in particular, the disulfide bond (-SS-) between the ligand chains can be reduced to form a free sulfhydryl group; wherein the S atom in the compound represented by formula (Va) or formula (Vb) can be derived from the sulfhydryl group of Ab;

[0952] In the second step, the compound represented by any of the formulas (Va) or (Vb) of the present application is incubated in an alkaline buffer at a selected temperature for a certain period of time to obtain another compound represented by any of the formulas (Va) or (Vb) of the present application;

[0953] Tr, L2, L3 can be defined as Tr, L2, L3 in the compound represented by any formula (IVa) or formula (IVb) of the present application; Ab, N a-I Ab, N in the compound represented by any formula (Va) or formula (Vb) of the present application can be respectively a-I defined.

[0954] The buffer is selected from the following buffers with a pH of 2 to 12, citric acid-sodium citrate buffer, phosphoric acid-sodium phosphate buffer, phosphoric acid-potassium phosphate buffer, sodium dihydrogen phosphate-disodium hydrogen phosphate buffer, potassium dihydrogen phosphate-dipotassium hydrogen phosphate buffer, succinic acid-sodium succinate buffer, acetic acid-sodium acetate buffer, boric acid-borax buffer, boric acid-potassium borate buffer, borax-sodium hydroxide buffer, histidine-hydrochloric acid buffer, glycine-sodium hydroxide buffer, arginine-hydrochloric acid buffer, sodium bicarbonate-sodium carbonate buffer, potassium bicarbonate-potassium carbonate buffer, Tris-hydrochloric acid buffer, ammonia-ammonium chloride buffer, sodium barbital-hydrochloric acid buffer, borax-sodium carbonate buffer, boric acid-potassium chloride buffer, or a combination of two or more of the above buffers; the selected temperature can be 5 to 60° C.; and the incubation time can be 0 to 72 hours.

[0955] Preparation Route 20

[0956]

[0957] The ligand Ab and a compound having a group capable of coupling with two thiol groups represented by any of formula (IVa) or formula (IVb) of the present application are reacted in an acidic, neutral or alkaline buffer to obtain a compound represented by formula (Va) or formula (Vb);

[0958] wherein: Ab is a ligand containing at least two free sulfhydryl groups (-SH), wherein the free sulfhydryl groups can be obtained by reducing the ligand with a reducing agent; the reducing agent includes but is not limited to tris(2-carboxyethyl)phosphine, mercaptoethanol, dithiothreitol, cysteine, reduced glutathione, etc.; in particular, the disulfide bonds (-SS-) between the ligand chains can be reduced to form free sulfhydryl groups; wherein the S atom in the compound represented by formula (Va) or formula (Vb) can be derived from the sulfhydryl groups of Ab;

[0959] Tr, L1, L2, L3 can be defined as Tr, L1, L2, L3 in the compound represented by any formula (IVa) or formula (IVb) of the present application; Ab, N a-I Ab, N in the compound represented by any formula (Va) or formula (Vb) of the present application can be respectively a-I Defined; L 1y Represents the structure after coupling with two thiol groups of L1.

[0960] The compound represented by formula (Va) or formula (Vb) having a group capable of coupling with two thiol groups may contain L selected from the following group: 1y Group: Each of the R L1a , R L1b and R L1cEach is independently selected from the following group: hydrogen, protium, deuterium, tritium, halogen, -NO2, -CN, -OH, -SH, -NH2, -C(O)H, -CO2H, -C(O)C(O)H, -C(O)CH2C(O)H, -S(O)H, -S(O)2H, -C(O)NH2, -SO2NH2, -OC(O)H, -N(H)SO2H, alkyl, alkenyl, alkynyl, alicyclic, heterocyclic, aryl and heteroaryl.

[0961] The buffer is selected from the following buffers with a pH of 2 to 12, citric acid-sodium citrate buffer, phosphoric acid-sodium phosphate buffer, phosphoric acid-potassium phosphate buffer, sodium dihydrogen phosphate-disodium hydrogen phosphate buffer, potassium dihydrogen phosphate-dipotassium hydrogen phosphate buffer, succinic acid-sodium succinate buffer, acetic acid-sodium acetate buffer, boric acid-borax buffer, boric acid-potassium borate buffer, borax-sodium hydroxide buffer, histidine-hydrochloric acid buffer, glycine-sodium hydroxide buffer, arginine-hydrochloric acid buffer, sodium bicarbonate-sodium carbonate buffer, potassium bicarbonate-potassium carbonate buffer, Tris-hydrochloric acid buffer, ammonia-ammonium chloride buffer, sodium barbital-hydrochloric acid buffer, borax-sodium carbonate buffer, boric acid-potassium chloride buffer, or a combination of two or more of the above buffers.

[0962] Example 1

[0963] Compound 1

[0964]

[0965] Step 1: Compound 1A (500 mg, 3.67 mmol) was dissolved in dichloromethane (10 mL), and DIEA (1.42 g, 10.99 mmol) was added. TBSCl (830 mg, 5.51 mmol) in dichloromethane (5 mL) was added to the reaction mixture and stirred overnight for 17 hours. TLC (PE / EA = 20 / 1) indicated the reaction was complete. The reaction mixture was directly dried under reduced pressure and filtered through a column chromatography (PE:EA = 100:0-100:1) to obtain 800 mg of 1B as a milky white oil (yield: 87%).

[0966] Step 2: Compound 1B (125 mg, 0.50 mmol) and Compound 1C (188 mg, 0.50 mmol) were dissolved in ultra-dry acetonitrile (8 mL), replaced with nitrogen, and cooled to 0°C. Methanesulfonic acid (48 mg, 0.50 mmol) and MeCN (2 mL) were added via syringe and stirred overnight for 16 hours. TLC (PE / EA = 3 / 1) indicated the reaction was complete. Sodium bicarbonate was added, filtered, and dried to give the crude product, which was then subjected to preparative purification to afford 62 mg of Compound 1 as a white powder and 7 mg of Compound 1S as a white powder, in yields of 25% and 3%, respectively.

[0967] Compound 1:

[0968] MS-ESI: m / z 495.3[M+H] + .

[0969] 1 H NMR(400MHz,DMSO-d6)δ7.41(d,J=8.1Hz,2H),7.34-7.28(m,3H),6.16(dd,J=10.1,1.9Hz ,1H),5.93(s,1H),5.43(s,1H),4.93(d,J=4.9Hz,1H),4.79(brs,1H),4.52(d,J=19.5Hz, 1H),4.48(s,2H),4.34-4.25(m,1H),4.19(d,J=19.5Hz,1H),2.61-2.52(m,1H),2.35-2.27 (m,1H),2.19-1.98(m,2H),1.84-1.58(m,5H),1.39(s,3H),1.13-0.95(m,2H),0.87(s,3H).

[0970] Compound 1S:

[0971] MS-ESI: m / z 495.3[M+H] + .

[0972] 1 H NMR (400MHz, DMSO-d6) δ7.35-7.27(m,3H),7.22(d,J=8.1Hz,2H),6.17(dd,J=10.1,1.9Hz,1H),6 .09(s,1H),5.95(s,1H),5.30(d,J=6.8Hz,1H),4.81-4.76(m,1H),4.48(s,2H),4.30(s,1H),4.25 (d,J=19.2Hz,1H),4.01(d,J=19.2Hz,1H),2.59-2.53(m,1H),2.36-2.28(m,1H),2.12-1.97(m,2 H),1.90-1.69(m,5H),1.39(s,3H),1.26-1.13(m,1H),1.06(dd,J=11.0,3.4Hz,1H),0.89(s,3H).

[0973] Compound 2

[0974]

[0975] Step 1: Compound 2A (5.0 g, 25.4 mmol) and imidazole (2.59 g, 38.0 mmol) were dissolved in dichloromethane (50 mL). TBSCl (4.9 g, 32.5 mmol) was added and allowed to react at 25°C for 2 hours. TLC indicated the reaction was complete. The mixture was extracted twice with 100 mL of water and 100 mL of dichloromethane. The combined dichloromethanes were dried over anhydrous sodium sulfate and then spin-dried. The mixture was directly passed through a column and purified by PE:EA (20:1) to obtain 3 g of a yellow solid 2B (yield: 38%). MS-ESI: m / z 312.2 [M+H] + .

[0976] Step 2: Compound 2B (1.0 g, 3.2 mmol) was dissolved in THF (10 mL). After N2 substitution, the temperature was lowered to 0°C and BH3 / THF (6.4 mL, 6.4 mmol) was added. After 17 hours, LCMS indicated the reaction was complete. The reaction solution was added to methanol (40 mL) (maintained at 0°C in an ice bath), the BH3 was quenched, and the mixture was dried to obtain 500 mg of a colorless oil, 2C, in a 52% yield. MS-ESI: m / z 298.1 [M+H] + .

[0977] Step 3: Compound 2C (500 mg, 1.68 mmol) was dissolved in dichloromethane (8 mL). The mixture was cooled to 0°C and Dess-Martin reagent (1.43 g, 3.36 mmol) was added. After 1.5 hours, LCMS indicated that the reaction was complete. The reaction solution was diluted with water (100 mL) and extracted twice with ethyl acetate (100 mL + 50 mL). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and then spin-dried to obtain 250 mg of a white solid 2D (yield: 50%). MS-ESI: m / z 296.0 [M+H] + .

[0978] Step 4: Compound 2D (100 mg, 0.34 mmol), compound 1C (127 mg, 0.34 mmol), and MgSO4 (250 mg, 2.08 mmol) were dissolved in acetonitrile (10 mL). The mixture was cooled to 0°C and purged with nitrogen. Trifluoromethanesulfonic acid (150 mg, 1.0 mmol) was added and the mixture was kept in an ice bath for 2 hours. LCMS indicated the reaction was complete. The reaction solution was filtered directly, and the filter cake was rinsed three times with acetonitrile. The mother liquor was then dried and purified by column chromatography (DCM:MeOH = 30:1) to afford 100 mg of 2E as a white solid in a 55% yield. MS-ESI: m / z 540.0 [M+H] + .

[0979] Step 5: Compound 2E (100 mg, 0.18 mmol) and iron powder (100 mg, 1.8 mmol) were added to ethanol (4 mL). NH4Cl (96 mg, 1.8 mmol) was dissolved in water and added to the ethanol. The mixture was replaced with N2 and heated to 60°C for 2 hours. LCMS showed that the reaction was complete. The reaction solution was filtered through a pad of celite, the filter cake was rinsed three times with ethanol, and the mother liquor was dried and sent to the preparation. After freeze-drying, 40 mg of white solid powder 2 and 5 mg of white solid powder 2S were obtained, with yields of 42% and 5%, respectively.

[0980] Compound 2:

[0981] MS-ESI: m / z 532.2[M+Na] + .

[0982] 1 H NMR (400MHz, DMSO-d6) δ7.32(d,J=10.1Hz,1H),7.20(d,J=7.7Hz,1H),6.94(s,1H),6.89(d,J=7.7Hz,1H),6.16 (dd,J=10.1,1.9Hz,1H),5.93(s,1H),5.34(s,1H),4.95-4.89(m,1H),4.80(brs,1H),4.49(d,J=19.4Hz,1H),4. 44(s,2H),4.33-4.27(m,1H),4.18(d,J=19.4Hz,1H),2.62-2.52(m,1H),2.35-2.28(m,1H),2.19-2.07(m,1H),2 .07-1.99(m,1H),1.80-1.60(m,5H),1.40(s,3H),1.15-1.02(m,1H),0.99(dd,J=11.2,3.6Hz,1H),0.86(s,3H).

[0983] Compound 2S:

[0984] MS-ESI: m / z 532.3[M+Na] + .

[0985] 1H NMR(400MHz, DMSO-d6)δ7.32(d,J=10.1Hz,1H),7.15-7.09(m,1H),6.70(s,1H),6.63(d,J=7.6Hz, 1H),6.17(dd,J=10.0,1.9Hz,1H),5.99(s,1H),5.94(s,1H),5.25(d,J=6.8Hz,1H),4.78(brs,1H), 4.40(s,2H),4.33-4.22(m,2H),4.02(d,J=19.2Hz,1H),2.60-2.53(m,1H),2.36-2.28(m,1H),2.1 1-1.97(m,2H),1.90-1.69(m,5H),1.39(s,3H),1.26-1.13(m,1H),1.09-1.02(m,1H),0.88(s,3H).

[0986] Compound 3

[0987]

[0988] Step 1: Compound 3A (6.58 g, 33.08 mmol, 1.0 eq) and compound 3B (5.03 g, 33.08 mmol, 1.0 eq) were placed in a 250 mL three-necked flask. THF (50 mL), water (5 mL), and KCO (13.71 g, 99.24 mmol, 3.0 eq) were then added. Pd(dppf)Cl (1.21 g, 1.65 mmol, 0.05 eq) was then added under nitrogen. The atmosphere was replaced with nitrogen three times. The mixture was heated to 80°C and refluxed. After stirring for 3-4 hours, TLC (PE / EA = 5 / 1) indicated the reaction was complete. The reaction solution was cooled to room temperature and filtered. The filtrate was poured into 300 mL of water and extracted with EA (200 mL). The organic phase was washed with water (100 mL × 3), washed once with saturated NaCl, dried over anhydrous Na2SO4, and dried by spin drying. The sample was mixed with silica gel and the product was purified by column chromatography (PE / EA = 7 / 1). The product was dried by spin drying to obtain 2.5 g of white solid 3C, with a yield of 33%.

[0989] Step 2: Compound 1D (3 g, 7.97 mmol, 1.0 eq), compound 3C (1.8 g, 7.97 mmol, 1.0 eq), and MgSO4 (2.88 g, 23.91 mmol, 3.0 eq) were placed in a 100 mL three-necked flask. Acetonitrile (30 mL) was added and, under nitrogen protection, trifluoromethanesulfonic acid (1.2 g, 7.97 mmol, 1.0 eq) was added under an ice-water bath. After addition, the mixture was warmed to room temperature and reacted for 2 hours. TLC (PE / EA = 3 / 1) indicated completion of the reaction. The reaction solution was filtered, the filtrate was spin-dried, and the product was purified by silica gel column chromatography (PE / EA = 2 / 1). The product was spin-dried to give 3.7 g of a white solid 3, in a 79% yield.

[0990] MS-ESI: m / z 585.3[M+H] + .

[0991] 1 H NMR (400MHz, DMSO-d6) δ7.36(d,J=7.8Hz,2H),7.30(d,J=10.1Hz,1H),7.24(d,J=7.7Hz,2H),7.20(d,J=7.5Hz,1H),7.15 (s,1H),7.11(d,J=7.6Hz,1H),7.07(d,J=7.6Hz,1H),6.16(dd,J=10.1,1.9Hz,1H),5.92(s,1H),5.39(s,1H),4.91(d,J= 4.8Hz,1H),4.78(brs,1H),4.49(d,J=19.4Hz,1H),4.43(s,2H),4.32-4.25(m,1H),4.17(d,J=19.5Hz,1H),3.90(s,2H), 2.59-2.51(m,1H),2.35-2.26(m,1H),2.18-1.95(m,2H),1.82-1.56(m,5H),1.39(s,3H),1.11-0.96(m,4H),0.85(s,3H).

[0992] Compound 4

[0993]

[0994] Step 1: Compound 4A (500 mg, 2.51 mmol) was dissolved in THF / H₂O (10 / 2 mL), followed by the addition of compound 4B (573 mg, 3.77 mmol) and K₂CO₃ (1.04 g, 7.53 mmol). After nitrogen was replaced, Pd(dppf)Cl₂ (367 mg, 0.50 mmol) was added and the mixture was heated to 80°C under nitrogen for 2 hours. The reaction mixture was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (PE / EA = 1:0 to 3:1) to afford 270 mg of 4C as a yellow solid (yield: 48%).

[0995] Step 2: Compound 4C (150 mg, 0.66 mmol) was dissolved in MeCN (10 mL), followed by the addition of compound 1C (250 mg, 0.66 mmol) and MgSO4 (160 mg, 1.32 mmol). Trifluoromethanesulfonic acid (299 mg, 1.99 mmol) was added under nitrogen at 0°C. The reaction was continued at 0°C for 1 hour. The reaction solution was diluted with water (20 mL) and extracted with dichloromethane (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL), dried, and then spin-dried to obtain a crude product. 120 mg of a white powdery solid 4 and 13 mg of a white powdery solid 4S were obtained by the preparation method, with yields of 31% and 3%, respectively.

[0996] Compound 4:

[0997] MS-ESI: m / z 585.4[M+H] + .

[0998] 1 H NMR (400MHz, DMSO-d6) δ7.35(d,J=8.0Hz,2H),7.30(d,J=10.1Hz,1H),7.24-7.16(m,4H),7.14(d,J=8 .1Hz,2H),6.16(dd,J=10.1,1.9Hz,1H),5.92(s,1H),5.38(s,1H),4.90(d,J=4.8Hz,1H),4.48(d,J=1 9.4Hz,1H),4.42(s,2H),4.31-4.25(m,1H),4.16(d,J=19.4Hz,1H),3.89(s,2H),2.59-2.52(m,1H),2 .36-2.25(m,1H),2.18-1.96(m,2H),1.82-1.55(m,5H),1.38(s,3H),1.10-0.94(m,2H),0.85(s,3H).

[0999] Compound 4S:

[1000] MS-ESI: m / z 585.3[M+H] + .

[1001] 1 H NMR(400MHz,Chloroform-d)δ7.31-7.26(m,3H),7.21-7.11(m,6H),6.31(dd,J=10.1,1.9Hz, 1H),6.08(s,2H),5.43-5.38(m,1H),4.65(s,2H),4.52-4.45(m,1H),4.27(d,J=19.9Hz,1H),4 .05(d,J=19.9Hz,1H),3.95(s,2H),2.66-2.54(m,1H),2.23-2.07(m,3H),1.95-1.83(m,2H), 1.80-1.68(m,1H),1.63(dd,J=14.1,2.6Hz,1H),1.46(s,3H),1.31-1.13(m,2H),0.99(s,3H).

[1002] Compound 5

[1003]

[1004] Step 1: Compound 5A (50 g, 216 mmol, 1.0 eq) and imidazole (22 g, 323 mmol, 1.5 eq) were added to a 2 L three-necked flask and dissolved in dichloromethane (700 mL). TBSCl (44 g, 292 mmol, 1.35 eq) was then slowly added. TLC (PE / EA = 9 / 1) confirmed the completion of the reaction. The reaction solution was cooled to room temperature and filtered. The product was extracted twice with dichloromethane (100 mL) and H₂O (90 mL). The organic phase was dried over anhydrous sodium sulfate, dried, and purified by a normal phase column (PE:EA = 10:1) to obtain 65 g of 5B as a white solid in an 87% yield.

[1005] Step 2: Compound 5B (60 g, 173 mmol, 1.0 eq), pinacol diboronate (44 g, 173 mmol, 1.0 eq), Pd(dppf)Cl2 (6.3 g, 8.6 mmol, 0.05 eq), and potassium acetate (51 g, 520 mmol, 3.0 eq) were added to a 2 L three-necked flask and dissolved in 1 L of dioxane. Under N2 protection, the mixture was stirred at 110°C for 12 hours. TLC (PE:EA = 9:1) indicated the reaction was complete. The reaction solution was cooled to room temperature and filtered. The mixture was extracted twice with ethyl acetate (1 L) and H2O (1 L). The organic phase was dried over anhydrous sodium sulfate, dried, and purified by a normal phase column (PE:EA = 10:1) to obtain 60 g of 5C as a white solid in an 88% yield.

[1006] Step 3: Compound 5C (30 g, 76 mmol, 1.0 eq), compound 5D (15.2 g, 76 mmol, 1 eq), and Pd(dppf)Cl2 (3 g, 4 mmol, 0.05 eq) were added to a 1 L three-necked flask, followed by the addition of 600 mL of dioxane and Cs2CO3 (74 g, 228 mmol, 3.0 eq) dissolved in 60 mL of water. The mixture was stirred at 110° C. for 2 hours under nitrogen protection. TLC (PE:EA=9:1) indicated the reaction was complete. The reaction solution was cooled to room temperature and filtered. The mixture was extracted twice with ethyl acetate (1 L) and H2O (1 L). The organic phase was dried over anhydrous sodium sulfate, dried, and purified by a normal phase column (PE:EA=10:1) to obtain 20 g of 5E as a white solid in a yield of 68%.

[1007] Step 4: Compound 5E (2.0 g, 5.2 mmol, 1.0 eq), compound 1C (1.96 g, 5.2 mmol, 1.0 eq), and MgSO4 (1.88 g, 15.6 mmol, 3.0 eq) were placed in a 250 mL three-necked flask. 50 mL of acetonitrile was added and cooled to -20°C with dry ice ethanol. Trifluoromethanesulfonic acid (2.34 g, 15.6 mmol, 3.0 eq) was slowly added and stirred at 0°C for 2 hours. TLC (PE:EA = 1:1) showed that the reaction was complete. Sodium bicarbonate solution was added to the reaction solution to adjust the pH to 7-8. The acetonitrile was evaporated completely, extracted, and slurried with PE:EA = 3:1 to obtain 3.2 g of crude compound 5F (yield: 98%), which was used directly in the next reaction. MS-ESI: m / z 630.1 [M+H] + .

[1008] Step 5: Compound 5F (3.2 g, 5.08 mmol, 1 eq) and Fe (2.84 g, 50.8 mmol, 10 eq) were placed in a 250 mL three-necked flask. 80 mL of ethanol was added. NH4Cl (2.72 g, 50.8 mmol, 10 eq) dissolved in 80 mL of water was added to the flask and reacted at 60°C for 2 hours. TLC (DCM:MOH = 10:1) showed the reaction was complete. The ethanol was evaporated under reduced pressure, the pH was adjusted to 8 with aqueous sodium bicarbonate solution, and the mixture was extracted twice with 80 mL of dichloromethane and water. The organic phase was dried over magnesium sulfate and filtered. The organic phase was spin-dried to obtain 2.6 g of a yellow solid. 400 mg of the crude product was removed and passed through a reverse phase column to obtain 43.9 mg of compound 5 with a purity of more than 95%.

[1009] MS-ESI: m / z 600.1[M+H] + .

[1010] 1 H NMR (400MHz, DMSO-d6) δ7.38(d,J=8.0Hz,2H),7.31(d,J=10.1Hz,1H),7.24(d,J=7.9Hz,2H),6.96(s,1H),6.9 0(s,1H),6.76(s,1H),6.16(dd,J=10.1,1.8Hz,1H),5.93(s,1H),5.40(s,1H),4.92(d,J=4.8Hz,1H),4.79(brs ,1H),4.49(d,J=19.4Hz,1H),4.43(s,2H),4.32-4.26(m,1H),4.17(d,J=19.5Hz,1H),3.90(s,2H),2.62-2.52 (m,1H),2.36-2.26(m,1H),2.18-1.97(m,2H),1.81-1.55(m,5H),1.39(s,3H),1.10-0.94(m,2H),0.86(s,3H).

[1011] Compound 6

[1012]

[1013] Step 1: Compound 6A (10 g, 43.1 mmol) was dissolved in dichloromethane (100 mL). DIEA (16.7 g, 129.3 mmol) was added. TBSCl (9.7 g, 64.6 mmol) in dichloromethane (100 mL) was added to the reaction mixture and stirred overnight for 3 days. TLC (PE / EA = 40 / 1) indicated the reaction was complete. The reaction mixture was directly dried under reduced pressure and filtered through a column with pure petroleum ether to obtain 12.0 g of 6B as a light yellow oil (yield: 80%).

[1014] Step 2: Compound 6B (1 g, 2.89 mmol), pinacol diboronate (883 mg, 3.48 mmol), 1,4-dioxane (10 mL), KOAc (853 mg, 8.69 mmol), and Pd(dppf)Cl2 (212 mg, 0.29 mmol) were dissolved in 1,4-dioxane (10 mL), purged with nitrogen, and heated (110°C) with stirring overnight for 17 hours. TLC (PE:EA = 40:1) indicated the reaction was complete. The reaction solution was directly spin-dried and the sample was passed through a column chromatography column. 183 mg of 6C was obtained from a 50:1 mixture of petroleum ether and ethyl acetate (yield: 16%) as a light yellow oil. MS-ESI: m / z 394.3 [M+H] + .

[1015] Step 3: Compound 6C (180 mg, 0.46 mmol), Pd(dppf)Cl2 (33.5 mg, 0.046 mmol), compound 5D (90.6 mg, 0.46 mg), Cs2CO3 (298 mg, 0.92 mmol), 1,4-dioxane (8 mL), and water (2 mL). The atmosphere was replaced with N2, heated (110°C), and stirred overnight (20 hours). TLC (PE / EA = 10 / 1) indicated the reaction was complete. The reaction solution was directly spin-dried and the sample was passed through a column chromatography column. 95.8 mg of 6D was obtained as a light yellow oily liquid from a 100:1 mixture of petroleum ether and ethyl acetate, with a yield of 54%. MS-ESI: m / z 386.1 [M+H] + .

[1016] Step 4: Compound 6D (500 mg, 1.30 mmol), compound 1C (488 mg, 1.30 mmol), MeCN (10 mL), and MgSO4 (930 mg, 7.73 mmol) were placed in a three-necked flask. The atmosphere was replaced with N2 and cooled to 0°C. Trifluoromethanesulfonic acid (585 mg, 3.90 mmol) was added using a constant pressure liquid funnel and stirred for 3 hours. TLC (DCM / MeOH = 10 / 1) indicated the reaction was complete. The reaction solution was directly spin-dried and the sample was passed through a column chromatography column. 300 mg of 6E was obtained as a white solid from a 50:1 dichloromethane:MeOH mixture, yield 37%. MS-ESI: m / z 630.1 [M+H] + .

[1017] Step 5: Compound 6E (4 g, 6.35 mmol) and ethanol (100 mL) were added to a single-necked flask. Iron powder (2.36 g, 42.1 mmol) and NH4Cl (3.4 g, 63.6 mmol) were added and dissolved in water (40 mL). The mixture was then added to the above single-necked flask, the atmosphere was replaced with nitrogen, and the mixture was heated and stirred at 60°C for 2 hours. TLC (dichloromethane / CH3OH = 10 / 1) indicated the reaction was complete. Aqueous NaHCO3 solution was added to adjust the pH to between 7 and 8, stirred for 10 minutes, and filtered. The filtrate was then dried and dissolved in dichloromethane and stirred before column chromatography. 3.5 g of white powder 6 was obtained from a 30:1 mixture of dichloromethane and methanol, yield: 92%. 50 mg of the white powder was filtered, dried, and lyophilized after addition of acetonitrile and water (purified water) to obtain 38 mg of product 6 with a purity of 90.08%.

[1018] MS-ESI: m / z 622.3[M+Na] + .

[1019] 1 H NMR (400MHz, DMSO-d6) δ7.35(d,J=8.0Hz,2H),7.30(d,J=10.2Hz,1H),7.22-7.16(m,2H),6.93(d,J=7.6Hz,1H),6. 45-6.40(m,1H),6.40-6.34(m,1H),6.16(dd,J=10.1,1.9Hz,1H),5.93(s,1H),5.39(s,1H),5.14-5.04(m,1H),4.9 8-4.83(m,3H),4.78(d,J=3.4Hz,1H),4.54-4.45(m,1H),4.34-4.26(m,3H),4.22-4.12(m,1H),3.75(s,2H),2.60- 2.53(m,1H),2.36-2.26(m,1H),2.17-1.96(m,2H),1.81-1.60(m,5H),1.39(s,3H),1.09-0.97(m,2H),0.85(s,3H).

[1020] Compound 7

[1021]

[1022] Step 1: Compound 7A (65 g, 280 mmol, 1.0 eq) and imidazole (29 g, 420 mmol, 1.5 eq) were added to a 2 L three-necked flask and dissolved in dichloromethane (700 mL). TBSCl (59 g, 392 mmol, 1.4 eq) was then slowly added. TLC (PE:EA = 9:1) confirmed the complete reaction. The reaction solution was cooled to room temperature and filtered. The product was extracted twice with dichloromethane (100 mL) and H2O (90 mL). The organic phase was dried over anhydrous sodium sulfate, dried, and purified by a normal phase column (PE:EA = 10:1) to obtain 80 g of 7B as a white solid (yield: 82%).

[1023] Step 2: Compound 7B (70 g, 203 mmol, 1.0 eq), bis(pyrazolyl)boronic acid (51 g, 203 mmol, 1.0 eq), Pd(dppf)Cl2 (7.4 g, 10.1 mmol, 0.05 eq), and potassium acetate (60 g, 609 mmol, 3.0 eq) were added to a 2 L three-necked flask. 1 L of dioxane was added for dissolution. Under N2 protection, the mixture was stirred at 110°C for 12 hours. TLC (PE:EA = 10:1) indicated the reaction was complete. The reaction solution was cooled to room temperature and filtered. The mixture was extracted twice with ethyl acetate (1 L) and H2O (1 L). The organic phase was dried over anhydrous sodium sulfate, dried, and purified by normal phase column chromatography (PE:EA = 10:1) to obtain 60 g of 7C as a white solid in a 75% yield.

[1024] Step 3 Compound 7C (15 g, 38 mmol, 1.0 eq), compound 5D (7.6 g, 38 mmol, 1 eq), and Pd(dppf)Cl2 (1.4 g, 1.9 mmol, 0.05 eq) were added to a 1 L three-necked flask, followed by the addition of 600 mL of dioxane and Cs2CO3 (37 g, 114 mmol, 3.0 eq) dissolved in 50 mL of water. The mixture was stirred at 110° C. for 2 hours under nitrogen protection. TLC (PE:EA=10:1) indicated the reaction was complete. The reaction solution was cooled to room temperature and filtered. The mixture was extracted twice with ethyl acetate (1 L) and H2O (1 L). The organic phase was dried over anhydrous sodium sulfate, dried, and purified by a normal phase column (PE:EA=10:1) to obtain 10 g of 7D as a white solid in a yield of 68%.

[1025] Step 4: Compound 7D (1 g, 2.6 mmol, 1.0 eq), compound 1C (0.98 g, 2.6 mmol, 1.0 eq), and MgSO4 (0.94 g, 7.8 mmol, 3.0 eq) were placed in a 250 mL three-necked flask. 50 mL of acetonitrile was added and the mixture was cooled to -20°C with dry ice ethanol. Trifluoromethanesulfonic acid (1.17 g, 7.8 mmol, 3.0 eq) was slowly added and stirred at 0°C for 2 hours. TLC (dichloromethane:MeOH = 10:1) indicated that the reaction was complete. Sodium bicarbonate solution was added to adjust the pH to 7-8. The acetonitrile was evaporated completely, extracted, and slurried with PE:EA = 3:1 to obtain 1.4 g of compound 7E, in an 86% yield. MS-ESI: m / z 630.1 [M+H] + .

[1026] Step 5: Compound 7E (200 mg, 0.317 mmol, 1 eq) and iron powder (177 mg, 3.17 mmol, 10 eq) were placed in a 25 mL three-necked flask. 6 mL of ethanol was added. NH4Cl (170 mg, 3.17 mmol, 10 eq) was dissolved in 6 mL of water and added to the flask. The mixture was reacted at 60°C for 2 hours. TLC (DCM:MeOH = 10:1) showed that the reaction was complete. The ethanol was evaporated under reduced pressure, the pH was adjusted to 8 with aqueous sodium bicarbonate solution, and the mixture was extracted twice with 80 mL of dichloromethane and water. The organic phase was dried over magnesium sulfate and filtered. The organic phase was spin-dried to obtain 200 mg of a yellow solid, which was then filtered through a reverse phase column to obtain 42.6 mg of compound 7, a yield of 22%.

[1027] MS-ESI: m / z 600.8[M+H] + .

[1028] 1H NMR (400MHz, DMSO-d6) δ7.36(d,J=8.0Hz,2H),7.31(d,J=10.1Hz,1H),7.22(d,J=7.9Hz,2H),7.20-7.14(m,1H) ,7.07(d,J=8.0Hz,1H),6.98(d,J=8.0Hz,1H),6.16(dd,J=10.1,1.9Hz,1H),5.93(s,1H),5.39(s,1H),4.91(d,J =4.7Hz,1H),4.79(s,1H),4.54-4.45(m,3H),4.31-4.26(m,1H),4.17(d,J=19.5Hz,1H),3.87(s,2H),2.60-2.5 3(m,1H),2.36-2.26(m,1H),2.18-1.96(m,2H),1.81-1.54(m,5H),1.39(s,3H),1.12-0.94(m,2H),0.86(s,3H).

[1029] Compound 8

[1030]

[1031] Step 1: Compound 3 (2 g, 3.42 mmol, 1.0 eq) and compound 8A (2.52 g, 6.84 mmol, 2.0 eq) were added to a reaction flask and dissolved in THF (30 mL). TsOH (0.63 g, 3.66 mmol, 1.07 eq) was added under nitrogen protection and allowed to react at 5-10°C for 30-40 minutes. TLC (PE / EA = 1 / 2) indicated completion of the reaction. The reaction solution was added to water, and the product was extracted with EA (100 mL). The organic phase was washed with water (50 mL x 3) and dried over anhydrous sodium sulfate. The sample was concentrated on silica gel and purified by column chromatography (PE / EA = 2 / 1). The product was then dried to give 2.3 g of 8B as a white solid, yield: 75%. MS-ESI: m / z 893.4 [M+H] + .

[1032] Step 2: Compound 8B (2 g, 2.24 mmol) was dissolved in DCM (20 mL). DEA (6 mL) was added dropwise under nitrogen in an ice-water bath. The mixture was allowed to react at room temperature for 2 hours. TLC (PE / EA = 1 / 2) indicated the reaction was complete. The reaction solution was spin-dried, washed three times with DCM, passed through a reverse-phase column, and lyophilized to obtain 600 mg of an off-white solid, 8C, in a 40% yield. MS-ESI: m / z 671.3 [M+H] + .

[1033] Step 3: Compound 8C (300 mg, 0.447 mmol, 1.0 eq) and compound 8D (230 mg, 0.492 mmol, 1.1 eq) were added to a reaction flask and dissolved in DMF (3.5 mL). Under nitrogen, DIEA (173 mg, 1.341 mmol, 3.0 eq) was added in an ice-water bath. A 1 mL DMF solution of HATU (221 mg, 0.581 mmol, 1.3 eq) was added dropwise. After addition, the mixture was stirred at 0-5°C for 30 minutes. LC-MS indicated the reaction was complete. The reaction solution was poured into ice water, and EA (100 mL) was added to extract the product. The organic phase was washed with water (50 mL x 3), dried over anhydrous sodium sulfate, and spin-dried to obtain 430 mg of a yellow solid 8E (yield: 86%). MS-ESI: m / z 1119.4 [M+H] + .

[1034] Step 4: Under nitrogen, compound 8E (300 mg, 0.268 mmol, 1 eq), Pd(PPh3)4 (62 mg, 0.0536 mmol, 0.2 eq), and N-methylmorpholine (452 ​​mg, 4.47 mmol, 16.7 eq) were added to a reaction flask. The mixture was reacted at room temperature for 1 hour under nitrogen. TLC indicated the reaction was complete. The reaction solution was directly purified by reverse-phase column chromatography to obtain 210 mg of 8F as a white solid in a 72% yield. MS-ESI: m / z 1079.3 [M+H] + .

[1035] Step 5: Under nitrogen, compound 8F (200 mg, 0.185 mmol) was dissolved in DCM (2.4 mL) and placed in an ice-water bath. DEA (0.8 mL) was added dropwise. After addition, the mixture was warmed to room temperature and reacted for 2 hours. LCMS indicated completion of the reaction. The reaction solution was directly extracted and washed three times with PE (60 mL x 3). The product precipitated and adhered to the wall of the vial. It was dissolved in acetonitrile (3 mL) and purified on a reverse-phase column to obtain 62 mg of 8G as a white solid, yield: 39%. MS-ESI: m / z 857.6 [M+H] + .

[1036] Step 6: Compound 8G (60 mg, 0.07 mmol, 1.0 eq) was dissolved in THF (3 mL), and water (0.6 mL) was added. The mixture was placed in an ice-water bath, and TEA (414 mg, 4.1 mmol, 58 eq) was added. The mixture was stirred for 1 minute, and bromoacetyl bromide (113 mg, 0.56 mmol, 8.0 eq) was added. The mixture was stirred at 0-5°C for 5 minutes. LC-MS showed that the reaction was complete. The reaction solution was stored in dry ice ethanol and sent to the preparation. After lyophilization, 11 mg of white solid 8 was obtained, with a yield of 16%.

[1037] MS-ESI: m / z 977.3[M+H] + .

[1038] 1 H NMR (400MHz, DMSO-d6) δ8.62(t,J=6.6Hz,1H),8.47(t,J=5.6Hz,1H),8.24(t,J=5.9Hz,1H),8.18(d,J=7.7Hz,1H),7.40-7.34(m,2H),7.3 1(d,J=10.1Hz,1H),7.27-7.20(m,3H),7.16(brs,1H),7.14-7.07(m,2H),6.16(dd,J=10.1,1.9Hz,1H),5.93(s,1H),5.39(s,1H),4.91(d, J=5.1Hz,1H),4.77(brs,1H),4.65-4.55(m,2H),4.49(d,J=19.4Hz,1H),4.39(s,2H),4.33-4.25(m,2H),4.17(d,J=19.5Hz,1H),3.94-3. 89(m,4H),3.80(d,J=5.6Hz,2H),3.72(d,J=5.8Hz,2H),2.57-2.52(m,2H),2.30-2.24(m,2H),2.18-2.06(m,1H),2.06-1.89(m,2H),1.83- 1.59(m,6H),1.39(s,3H),1.12-0.97(m,2H),0.85(s,3H).

[1039] Compound 9

[1040]

[1041] Step 1: Compound 4 (2.4 g, 4.1 mmol, 1.0 eq) and compound 8A (4.5 g, 12.3 mmol, 3.0 eq) were added to a reaction flask and dissolved in THF (30 mL). TsOH (1.1 g, 6.6 mmol, 1.6 eq) was added under nitrogen protection and allowed to react at 5-10°C for 30-40 minutes. TLC (PE / EA = 1 / 2) indicated completion of the reaction. The reaction solution was added to water, and the product was extracted with EA (100 mL). The organic phase was washed with water (50 mL × 3), dried over anhydrous sodium sulfate, concentrated on silica gel, and purified by column chromatography (PE / EA = 2 / 1). The product was then dried to obtain 1.5 g of 9A as a white solid, yield: 41%. MS-ESI: m / z 893.4 [M+H] + .

[1042] Step 2: Compound 9A (1.3 g, 1.46 mmol) was dissolved in DCM (13 mL). DEA (2.6 mL) was added dropwise under nitrogen in an ice-water bath. The mixture was allowed to react at room temperature for 2 hours. TLC (PE / EA = 1 / 2) indicated the reaction was complete. The reaction solution was spin-dried, washed three times with DCM, passed through a reverse-phase column, and lyophilized to obtain 350 mg of an off-white solid, 9B, in a 36% yield. MS-ESI: m / z 670.8 [M+H] + .

[1043] Step 3: Compound 9B (350 mg, 0.52 mmol, 1.0 eq) and compound 8D (243 mg, 0.52 mmol, 1.0 eq) were added to a reaction flask and dissolved in DMF (3.5 mL). Under nitrogen protection, DIEA (168 mg, 1.3 mmol, 2.5 eq) was added under an ice-water bath. A 1 mL DMF solution of HATU (240 mg, 0.63 mmol, 1.21 eq) was added dropwise. After addition, the mixture was stirred at 0-5°C for 30 minutes. LC-MS showed that the reaction was complete. The reaction solution was poured into ice water, EA (100 mL) was added, and the product was extracted. The organic phase was washed with water (50 mL × 3), dried over anhydrous sodium sulfate, and dried to obtain 600 mg of a foamy yellow solid 9C. The excess crude product was not counted in the yield. MS-ESI: m / z 1119.4 [M+H] + .

[1044] Step 4: Under nitrogen, compound 9C (400 mg, 0.35 mmol, 1.0 eq), Pd(PPh3)4 (82 mg, 0.071 mmol, 0.2 eq), and N-methylmorpholine (353 mg, 3.5 mmol, 10.0 eq) were added to a reaction flask. The mixture was reacted at room temperature for 1 hour under nitrogen. LC-MS indicated the reaction was complete. The reaction solution was directly purified by reverse phase column chromatography to obtain 150 mg of 9D as a white solid in a 39% yield. MS-ESI: m / z 1079.4 [M+H] + .

[1045] Step 5: Under nitrogen, compound 9D (150 mg, 0.14 mmol) was dissolved in DCM (1.5 mL) and placed in an ice-water bath. DEA (0.3 mL) was added dropwise. The mixture was warmed to room temperature and allowed to react for 2 hours. LCMS indicated the reaction was complete. The reaction solution was directly spin-dried, washed three times with DCM, and purified by reverse-phase column chromatography to obtain 60 mg of 9E as a white solid in a 50% yield. MS-ESI: m / z 857.4 [M+H] + .

[1046] Step 6: Compound 9E (50 mg, 0.058 mmol, 1.0 eq) was dissolved in THF (3 mL) and 3 drops of water were added to dissolve the mixture. The mixture was placed in an ice-water bath, and TEA (117 mg, 1.16 mmol, 20.0 eq) was added. The mixture was stirred for 1 minute. Bromoacetyl bromide (93 mg, 0.46 mmol, 8.0 eq) was added and stirred at 0-5°C for 5 minutes. LC-MS indicated the reaction was complete. The reaction solution was stored on dry ice and sent for preparation. After lyophilization, 11 mg of white solid 9 was obtained (yield: 19%).

[1047] MS-ESI: m / z 977.3[M+H] + .

[1048] 1 H NMR (400MHz, DMSO-d6) δ8.62(t,J=6.6Hz,1H),8.49(t,J=5.6Hz,1H),8.26(t,J=5.8Hz,1H),8.19(d,J=7.5Hz,1H),7.36(d,J=7.9Hz,2H),7.30(d ,J=10.1Hz,1H),7.26-7.14(m,6H),6.16(dd,J=10.1,1.9Hz,1H),5.93( s,1H),5.39(s,1H),4.91(d,J=4.9Hz,1H),4.79(brs,1H),4.61-4.54(m, 2H),4.49(d,J=19.4Hz,1H),4.37(s,2H),4.32-4.24(m,2H),4.16(d,J= 19.4Hz,1H),3.95-3.88(m,4H),3.79(d,J=5.5Hz,2H),3.72(d,J=5.8Hz, 2H),2.58-2.52(m,2H),2.30-2.24(m,2H),2.17-2.05(m,1H),2.05-1.89 (m,2H),1.82-1.58(m,6H),1.39(s,3H),1.11-0.96(m,2H),0.85(s,3H).

[1049] Compound 10

[1050]

[1051] Step 1: Compound 5 (1.9 g, 3.21 mmol, 1.0 eq), compound 8D (1.5 g, 3.21 mmol, 1.0 eq), and HATU (1.6 g, 4.24 mmol, 1.3 eq) were placed in a 100 mL three-necked flask. DMF (45 mL) was added to dissolve the mixture. 2,6-Lutidine (1.4 g, 12.72 mmol, 4.0 eq) was then slowly added. LCMS analysis indicated the reaction was complete. The reaction mixture was slowly added dropwise to 300 mL of water and filtered to obtain the product. The product was vacuum-dried using an oil pump to obtain 3.3 g of a yellow solid 10A, in a 94% yield. MS-ESI: m / z 1048.4 [M+H] + .

[1052] Step 2: Compound 10A (1.8 g, 1.72 mmol, 1.0 eq) and Pd(PPh3)4 (392 mg, 0.34 mmol, 0.2 eq) were added to a 50 mL three-necked flask and dissolved in dichloromethane (30 mL). Under nitrogen protection, N-methylmorpholine (868 mg, 8.6 mmol, 5.0 eq) was slowly added and allowed to react for half an hour. LCMS showed that the reaction was complete. Diethylamine (6 mL) was slowly added to the three-necked flask and stirred at room temperature for 1 hour. LCMS showed that the reaction was complete. 20 mL of DMF was added to the reaction solution and vacuum evaporated at room temperature until 20 mL of solvent remained. The vacuum evaporation was stopped and the solution was passed through a reverse phase column (ACN in water from 30% to 70%) and lyophilized to obtain 455 mg of a light yellow solid 10B (yield: 34%).

[1053] Step 3: Compound 10B (150 mg, 0.19 mmol, 1.0 eq) was dissolved in THF (5 mL) and water (0.2 mL), and triethylamine (190 mg, 1.9 mmol, 10 eq) was added. The mixture was cooled to 0°C in an ice-water bath. Bromoacetyl bromide (153 mg, 0.76 mmol, 4.0 eq) was dissolved in 1 mL of THF and then slowly added to the reaction solution. The mixture was reacted at 0°C for 15 minutes. LCMS showed that the reaction was complete, to give 20 mg of white solid 10, in a yield of 12%.

[1054] MS-ESI: m / z 906.3[M+H] + .

[1055] 1H NMR (400MHz, DMSO-d6) δ9.87(s,1H),8.52(s,1H),8.22(d,J=7.6Hz,1H),7.43(s,1H),7.38(d,J=7.9Hz,2H),7.31(d,J=9.7Hz,2 H),7.23(d,J=7.9Hz,2H),6.87(s,1H),6.16(d,J=10.3Hz,1H),5.93(s,1H),5.39(s,1H),4.91(d,J=5.1Hz,1H),4.78(s,1H),4.4 9(d,J=19.2Hz,1H),4.41(s,2H),4.39-4.32(m,1H),4.29(s,1H),4.17(d,J=19.5Hz,1H),3.93(s,2H),3.88(s,2H),3.82-3.77( m,2H),2.59-2.53(m,1H),2.35-2.21(m,4H),2.04-1.92(m,2H),1.84-1.63(m,6H),1.39(s,3H),1.09-0.99(m,2H),0.86(s,3H).

[1056] Compound 11

[1057]

[1058] Step 1: Compound 6 (300 mg, 0.5 mmol, 1.0 eq), compound 8D (233 mg, 0.5 mmol, 1.0 eq), and HATU (228 mg, 0.6 mmol, 1.2 eq) were placed in a 25 mL three-necked flask. After dissolution with DMF (5 mL), 2,6-lutidine (150 mg, 1.4 mmol, 2.8 eq) was slowly added. LCMS analysis indicated the reaction was complete. The reaction solution was slowly added dropwise to 100 mL of water and filtered to obtain the product. The product was vacuum-dried using an oil pump to obtain 450 mg of a yellow solid 11A, yield: 86%. MS-ESI: m / z 1048.4 [M+H] + .

[1059] Step 2: Compound 11A (450 mg, 0.43 mmol, 1.0 eq) and Pd(PPh3)4 (92 mg, 0.08 mmol, 0.2 eq) were placed in a 50 mL three-necked flask and dissolved in THF (10 mL). Under nitrogen, N-methylmorpholine (200 mg, 1.98 mmol, 4.6 eq) was slowly added. The mixture was allowed to react for half an hour. LCMS indicated completion of the reaction. The product was purified by a reverse phase column (ACN in water from 30% to 70%) and lyophilized to afford 250 mg of a pale yellow solid, 11B, in a 58% yield. MS-ESI: m / z 1008.3 [M+H] + .

[1060] Step 3: Compound 11B (250 mg, 0.25 mmol) was placed in a 25 mL three-necked flask and dissolved in DCM (10 mL). Under nitrogen, 2 mL of diethylamine was added and allowed to react for half an hour. LCMS indicated the reaction was complete. The product was purified by a reverse-phase column (ACN in water from 20% to 50%) and lyophilized to afford 130 mg of a pale yellow solid, 11C, in a 67% yield. MS-ESI: m / z 786.4 [M+H] + .

[1061] Step 4: Compound 11C (120 mg, 0.153 mmol, 1.0 eq) was dissolved in THF (5 mL) and water (0.2 mL). Triethylamine (155 mg, 1.53 mmol, 10.0 eq) was added, and the mixture was cooled to 0°C in an ice-water bath. Bromoacetyl bromide (123 mg, 0.61 mmol, 4.0 eq) was dissolved in 1 mL of THF and slowly added to the reaction mixture. The mixture was reacted at 0°C for 15 minutes. LCMS indicated the reaction was complete. Prep-HPLC analysis yielded 12.2 mg of 11 as a white solid (yield: 9%).

[1062] MS-ESI: m / z 906.3[M+H] + .

[1063] 1H NMR (400MHz, DMSO-d6) δ9.51(s,1H),8.58-8.45(m,1H),8.37(d,J=7.6Hz,1H),7.51-7.46(m,1H),7.40-7.34(m,2H),7.31(d,J=1 0.0Hz,1H),7.28-7.19(m,3H),7.01-6.94(m,1H),6.16(dd,J=10.0,1.9Hz,1H),5.93(s,1H),5.39(s,1H),4.91(d,J=5.0Hz,1H),4 .77(brs,1H),4.49(d,J=19.5Hz,1H),4.42(d,J=3.7Hz,1H),4.39-4.32(m,1H),4.29(brs,1H),4.17(d,J=19.5Hz,1H),3.98-3.77 (m,6H),2.60-2.52(m,2H),2.34-2.28(m,2H),2.18-1.96(m,3H),1.88-1.55(m,6H),1.39(s,3H),1.13-0.98(m,2H),0.85(s,3H).

[1064] Compound 12

[1065]

[1066] Step 1: To a solution of compound 4C (10.0 g, 44.2 mmol) in DCM (70 mL) was added TEA (8.94 g, 88.39 mmol, 12.30 mL), the temperature was lowered to 0°C, MsCl (6.08 g, 53.0 mmol, 4.10 mL) was added to the reaction, and the mixture was stirred at 25°C for 12 hours. LCMS showed that the starting material had been consumed. DCM (50 mL) was added to dilute the reaction solution and quenched with water (200 mL). The mixture was extracted with DCM (100 mL × 3), the organic phases were combined, washed with saturated brine (100 mL), and dried over anhydrous Na2SO4. The mixture was filtered and the filtrate was dried to give compound 12A (8.60 g, crude product) as a yellow oil.

[1067] Step 2: To a solution of compound 12A (8.60 g, 31.58 mmol) in DMF (51.6 mL) was added thioglycolic acid (7.21 g, 63.15 mmol) and stirred at 25°C for 5 hours. TLC (PE / EA = 5:1) showed that the starting material had reacted completely. Water (100 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phases were combined, washed with saturated brine, and then washed with a 5% LiCl aqueous solution. The organic phase was dried over anhydrous Na2SO4, filtered, and the filtrate was dried to give compound 12B (5.75 g, yield: 57.82%, purity: 90.2%) as a brown oil. MS-ESI: m / z 285.2 [M+H] + .

[1068] Step 3: Compound 12B (12.0 g, 42.2 mmol) was dissolved in methanol (240 mL), and K2CO3 (8.75 g, 63.3 mmol) was added. The mixture was stirred at 20°C for 3 hours. LCMS showed that the starting material had been completely converted. The reaction solution was poured into DCM (240 mL), and 0.5 M HCl (100 mL) was added. The liquid was separated, and the organic phase was dried over anhydrous sodium sulfate. After concentration, an off-white solid 12C (10.0 g, yield: 97.7%) was obtained. MS-ESI: m / z 483.2 [M+H] + .

[1069] Step 4: Compound 12C (9.00 g, 37.1 mmol), compound 1C (13.9 g, 37.1 mmol) and anhydrous magnesium sulfate (13.4 g, 111 mmol) were added to acetonitrile (270 mL). TfOH (16.7 g, 111 mmol, 9.84 mL) was added dropwise at 0°C. After the addition, the temperature was gradually raised to 20°C and stirred for 3 hours. LCMS showed that the starting material had been completely converted. The reaction solution was quenched with saturated aqueous sodium bicarbonate solution (60 mL) and extracted with ethyl acetate (100 mL × 2). The organic phase was washed with saturated brine (80 mL) and dried over anhydrous sodium sulfate. After concentration, an off-white solid 12D (15.0 g, yield: 33.6%) was obtained. MS-ESI: m / z 1199.4 [M+H] + .

[1070] Step 5: Compound 12D (290 mg, crude) was added to DMF (4 mL), followed by DL-dithiothreitol (200 mg, 1.30 mmol), and the mixture was allowed to react at room temperature under nitrogen for 16 hours. LCMS indicated the reaction was complete. The reaction solution was directly purified by HPLC to afford 12 as a white powdery solid (20 mg, yield: 13.8%). MS-ESI: m / z 601.2 [M+H] + .

[1071] 1 H NMR (400MHz, DMSO) δ7.37(d,J=8.1Hz,2H),7.31(d,J=10.1Hz,1H),7.23(dd,J=8.1,3.3Hz,4H),7.14(d,J=8.0Hz,2 H),6.16(dd,J=10.1,1.7Hz,1H),5.93(s,1H),5.39(s,1H),4.91(d,J=5.0Hz,1H),4.78(s,1H),4.49(d,J=19.4Hz,1 H),4.29(s,1H),4.17(d,J=19.5Hz,1H),3.88(d,J=11.3Hz,2H),3.67(d,J=7.6Hz,2H),2.77(t,J=7.6Hz,1H),2.50- 2.55(m,1H),2.45-2.25(m,1H),2.25-2.05(m,2H),1.80–1.53(m,5H),1.38(s,3H),1.14–0.94(m,2H),0.86(s,3H).

[1072] Compound 13

[1073]

[1074] Step 1 Compound 12 (7.00 g, 11.6 mmol) and compound 8A (4.29 g, 11.6 mmol) were added to DMF (49.0 mL), TfOH (3.50 g, 23.3 mmol, 2.06 mL) was added, and the mixture was stirred at 25 ° C for 16 hours. LCMS showed that the raw material had been completely converted. The reaction solution was quenched with saturated aqueous sodium bicarbonate solution (20 mL), extracted three times with ethyl acetate (100 mL×3), and the organic phase was washed with saturated brine (100 mL) and dried over anhydrous sodium sulfate. Concentration gave a yellow oil compound 13A (15.0 g, the crude product was not calculated in yield). MS-ESI: m / z 909.3 [M+H] + .

[1075] Step 2 Compound 13A (15.0 g, 16.5 mmol) was added to acetonitrile (105 mL), and diethylamine (6.03 g, 82.5 mmol, 8.50 mL) was added and stirred at 25°C for 16 hours. LCMS showed that the starting material had been completely converted. The reaction solution was slurried with acetonitrile (20.0 mL), filtered, and the resulting filter cake was drained. The crude product was prepared by reverse phase (water:acetonitrile = 25%-55%, 20 min) and lyophilized to obtain a white solid compound 13B (0.900 g, yield: 7.94%). MS-ESI: m / z 687.3 [M+H]+ .

[1076] Step 3: Compound 13B (500 mg, 727 μmol) and compound 8D (526 mg, 1.09 mmol) were added to DMF (5 mL), and HATU (830 mg, 2.18 mmol) and 2,6-lutidine (56.0 mg, 1.46 mmol, 169 μl) were added. The mixture was stirred at 20°C for 3 hours. LCMS showed that the starting material had been completely converted. The reaction solution was evacuated to obtain 550 mg of crude product. 250 mg of the crude product was prepared by reverse phase to obtain compound 13C (100 mg, yield: 26.2%) as a white solid. MS-ESI: m / z 1151.4 [M+H] + .

[1077] Step 4: Compound 13C (50.0 mg, 43.4 μmol) was added to acetonitrile (1 mL), followed by diethylamine (15.8 mg, 217 μmol, 22.3 μL), and the mixture was stirred at 25°C for 6 hours. LCMS showed complete conversion of the starting material. The reaction solution was spin-dried and slurried with methyl tert-butyl ether (3.00 mL) to afford Compound 13D (40.0 mg, yield: 99.1%) as a yellow solid. MS-ESI: m / z 929.2 [M+H] + .

[1078] Step 5: EEDQ (31.9 mg, 129 μmol) and bromoacetic acid (11.3 mg, 81.8 μmol) were dissolved in DMF (0.8 mL) and stirred at room temperature for 1 hour. Compound 13D (40.0 mg, 43.0 μmol) was added and stirred at 25°C for 2 hours. LCMS showed complete conversion of the starting material. After spin drying, the crude product was used directly in the next step to obtain a yellow solid, Compound 13E. MS-ESI: m / z 1049.2 [M+H] + .

[1079] Step 6: Compound 13E (45.0 mg, 42.8 μmol) was dissolved in DCM (1.50 mL), trifluoroacetic acid (770 mg, 6.75 mmol, 500 μL) was added, and the mixture was stirred at 25°C for 1 hour. LCMS showed that the starting material had been completely converted. After being spin-dried, the crude product was purified by reverse phase preparative purification to obtain compound 13 (3.06 mg, yield: 7.18%) as a yellow solid. MS-ESI: m / z 993.2 [M+H] + .

[1080] 1HNMR (400MHz, DMSO-d6) δ7.83(br d,1H,J=7.6Hz),7.63(br d,1H,J=7.2Hz),7.4(m,2H),7.26(br d,2H,J=8.0Hz),7.16(br d,1H,J=7.6Hz),4.92(br s,1H),4.48(brs,1H),4.30(br s,2H),4.1-4.3(m,2H),4.02(br d,1H,J=13.6Hz),3.92(br d,2H,J=8.4Hz),3.81(br d,2H,J=5.2Hz),3.74(br s,3H),3.63(br s,2H),2.91(s,2H),2.75(brs,1H),2.3-2.4(m,2H),2.27(br s,2H),2.1-2.1(m,1H),2.02(br s,1H),1.92(br s,1H),1.77(br s,3H),1.3-1.5(m,8H),1.25(s,2H),1.18(br d,1H,J=7.2Hz),1.02(br s,1H),0.8-1.0(m,2H).

[1081] Compound 14 (reference compound P1):

[1082]

[1083] It was synthesized according to patent CN109476699A.

[1084] Compound 15 (reference compound LP1):

[1085]

[1086] It was synthesized according to patent CN111465399A.

[1087] Compound 16 (reference compound LP2):

[1088]

[1089] It was synthesized according to patent CN111417410A.

[1090] Example 2

[1091] In this application, a 20 mM Histidine buffer solution at pH 5.5 is prepared by adjusting a 20 mM Histidine solution to pH 5.5 with HOAc. Stock solution 1: 10 mM EDTA buffer, pH 4.7; Stock solution 2: 800 mM Sodium citrate buffer, pH 8.4; Stock solution 3: 500 mM Urea solution; Stock solution 4: 10 mM tris(2-carboxyethyl)phosphine solution.

[1092] The protein purity of ADC in this application was detected by SEC method, and the parameters are shown below.

[1093]

[1094] The DAR value of the ADC in this application can be detected by LC-MS. The experimental procedure is as follows: 75.0 μL of 8.0 mol / L guanidine hydrochloride (Gdn-HCl), 5.0 μL of 1.0 mol / L Tris-HCl, and 2.0 μL of 1 mol / L DTT are added to a 40.0 μg ADC sample solution. The solution is diluted to 100.0 μL with ultrapure water, mixed thoroughly, and incubated at 22°C for 30 minutes. The drug / antibody ratio (DAR) is then analyzed using LC-MS. The LC parameters are shown below.

[1095]

[1096] The MS parameters are shown below.

[1097] instrument Agilent Technologies 6224 TOF LC / MS Ionization mode Positive Gas temperature 350℃ airflow 13L / min Sprayer pressure 45psig Capillary voltage 5000V Crushing voltage 250V Cone voltage 65V Peak-to-peak RF voltage on the octopole 750V Acquisition Mode MS(seg) Quality range 500-8000 m / z Acquisition rate 1spectra / s Collection time 1.4-7.0min

[1098] The DAR value of ADC in this application can also be detected by the HIC method, and the parameters are expressed as follows.

[1099]

[1100] Murine anti-TNFαmIgG2a 8c11: the light chain amino acid sequence is shown in SEQ ID NO: 31, and the heavy chain amino acid sequence is shown in SEQ ID NO: 32, prepared with reference to WO2015191783A2.

[1101] Antibody Adalimumab (Humira): The light chain amino acid sequence is shown in SEQ ID NO: 9, and the heavy chain amino acid sequence is shown in SEQ ID NO: 10, which were prepared with reference to US6090382A.

[1102] Antibody Litifilimab (BIIB059): The light chain amino acid sequence is shown in SEQ ID NO: 42, and the heavy chain amino acid sequence is shown in SEQ ID NO: 37, and was prepared with reference to CN105452295B.

[1103] Antibody Iscalimab: The light chain amino acid sequence is shown in SEQ ID NO: 19, and the heavy chain amino acid sequence is shown in SEQ ID NO: 20, which were prepared with reference to CN105949314B.

[1104] 2.1 Conjugate 1 (8c11-conjugate 1)

[1105]

[1106] To a solution of anti-mouse TNFαmIgG2a 8c11 in PB buffer (pH 6.5, 0.05 M PBS buffer; 2.5 ml, 9.96 mg / ml, 0.168 nmol) was added a prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37°C. The mixture was shaken in a water bath at 37°C for 3 hours to stop the reaction. The reaction solution was cooled to 25°C in a water bath, diluted to 5.0 mg / ml, and 2.0 ml of the solution was removed for further reaction.

[1107] Compound IV-1 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, added to the above 2.0 mL solution, and placed in a water bath shaker. The reaction was shaken at 25°C for 3 hours to stop the reaction. The reaction solution was desalted and purified using a Sephadex G25 gel column (elution phase: 0.05 M PBS buffer solution at pH 6.5, containing 0.001 M EDTA) to obtain an exemplary product of formula (Va) in PBS buffer (5.0 mg / mL, 1.1 mL).

[1108] The pH value of the ADC PBS buffer was adjusted to pH = 9 with 1M Tris buffer, incubated at 37°C for 24 hours, and then adjusted to pH 6.5 with 1mol / L citric acid buffer to obtain conjugate 1 (3.3mg / mL, 5.5ml). a-I Can be detected by LC-MS.

[1109] 2.2 Conjugate 2 (8c11-conjugate 2)

[1110]

[1111] To a PB buffered aqueous solution of antibody 8c11 (0.05 M PBS buffered aqueous solution, pH = 6.5; 2.5 ml, 9.96 mg / ml, 0.168 nmol) was added a prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37°C. The mixture was shaken in a water bath at 37°C for 3 hours to stop the reaction. The reaction solution was cooled to 25°C in a water bath, diluted to 5.0 mg / ml, and 2.0 ml of the solution was removed for further reaction.

[1112] Compound IV-12 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA and added to the above 2.0 mL solution. The mixture was placed in a water bath shaker and shaken at 25°C for 3 hours to stop the reaction. The reaction solution was desalted and purified using a Sephadex G25 gel column (elution phase: 0.05 M histidine buffer solution at pH 5.5, containing 0.001 M EDTA) to obtain conjugate 2 (5.0 mg / mL, 1.1 mL).

[1113] N a-I Can be detected by LC-MS.

[1114] 2.3 Conjugate 3 (8c11-conjugate 3)

[1115]

[1116] To a PB buffered aqueous solution of antibody 8c11 (0.05 M PBS buffered aqueous solution, pH = 6.5; 2.5 ml, 9.96 mg / ml, 0.168 nmol) was added a prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37°C. The mixture was shaken in a water bath at 37°C for 3 hours to stop the reaction. The reaction solution was cooled to 25°C in a water bath, diluted to 5.0 mg / ml, and 2.0 ml of the solution was removed for further reaction.

[1117] Compound IV-11 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, added to the above 2.0 mL solution, and placed in a water bath shaker. The reaction was shaken at 25°C for 3 hours, and the reaction was stopped. The reaction solution was desalted and purified using a Sephadex G25 gel column (elution phase: 0.05 M histidine buffer solution at pH 5.5, containing 0.001 M EDTA) to obtain conjugate 3 (5.3 mg / mL, 1.5 mL).

[1118] N a-I Can be detected by LC-MS.

[1119] 2.4 Conjugate 4 (8c11-conjugate 4)

[1120]

[1121] To a PB buffered aqueous solution of antibody 8c11 (0.05 M PBS buffered aqueous solution, pH = 6.5; 2.5 ml, 9.96 mg / ml, 0.168 nmol) was added a prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37°C. The mixture was shaken in a water bath at 37°C for 3 hours to stop the reaction. The reaction solution was cooled to 25°C in a water bath, diluted to 5.0 mg / ml, and 2.0 ml of the solution was removed for further reaction.

[1122] Compound IV-60 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, added to the above 2.0 mL solution, and placed in a water bath shaker. The reaction was shaken at 25°C for 3 hours, and the reaction was stopped. The reaction solution was desalted and purified using a Sephadex G25 gel column (elution phase: 0.05 M histidine buffer solution at pH 5.5, containing 0.001 M EDTA) to obtain conjugate 4 (4.7 mg / mL, 2.2 mL).

[1123] N a-I Can be detected by LC-MS.

[1124] 2.5 Conjugate 5 (8c11-conjugate 5)

[1125]

[1126] To a PB buffered aqueous solution of antibody 8c11 (0.05 M PBS buffered aqueous solution, pH = 6.5; 2.5 ml, 9.96 mg / ml, 0.168 nmol) was added a prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37°C. The mixture was shaken in a water bath at 37°C for 3 hours to stop the reaction. The reaction solution was cooled to 25°C in a water bath, diluted to 5.0 mg / ml, and 2.0 ml of the solution was removed for further reaction.

[1127] Compound IV-81 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, added to the above 2.0 mL solution, and placed in a water bath shaker. The reaction was shaken at 25°C for 3 hours, and the reaction was stopped. The reaction solution was desalted and purified using a Sephadex G25 gel column (elution phase: 0.05 M histidine buffer solution at pH 5.5, containing 0.001 M EDTA) to obtain conjugate 5 (2.9 mg / mL, 4.3 mL).

[1128] N a-I Can be detected by LC-MS.

[1129] 2.6 Conjugate 6 (8c11-conjugate 6)

[1130]

[1131] To a PB buffered aqueous solution of antibody 8c11 (0.05 M PBS buffered aqueous solution, pH = 6.5; 2.5 ml, 9.96 mg / ml, 0.168 nmol) was added a prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol) at 37°C. The mixture was shaken in a water bath at 37°C for 3 hours to stop the reaction. The reaction solution was cooled to 25°C in a water bath, diluted to 5.0 mg / ml, and 2.0 ml of the solution was removed for further reaction.

[1132] Compound IV-91 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA and added to the above 2.0 mL solution. The mixture was placed in a water bath shaker and shaken at 25°C for 3 hours to stop the reaction. The reaction solution was desalted and purified using a Sephadex G25 gel column (elution phase: 0.05 M histidine buffer solution at pH 5.5, containing 0.001 M EDTA) to obtain conjugate 6 (3.6 mg / mL, 5.5 mL).

[1133] N a-I Can be detected by LC-MS.

[1134] 2.7 Conjugate V-6

[1135]

[1136] At 37°C, a solution of antibody Adalimumab (Humira) in PB buffer (pH 6.5, 0.05 M PBS buffer; 2.5 ml, 9.96 mg / ml, 0.168 nmol) was added with a prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol). The mixture was placed in a water bath shaker and shaken at 37°C for 3 hours to stop the reaction. The reaction solution was cooled to 25°C in a water bath, diluted to 5.0 mg / ml, and 2.0 ml of the solution was removed for further reaction.

[1137] Compound IV-1 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, added to the above 2.0 mL solution, and placed in a water bath shaker. The reaction was shaken at 25°C for 3 hours to stop the reaction. The reaction solution was desalted and purified using a Sephadex G25 gel column (elution phase: 0.05 M PBS buffer solution at pH 6.5, containing 0.001 M EDTA) to obtain the exemplary product of formula (Va) in PBS buffer (2.8 mg / mL, 4.7 mL).

[1138] The pH value of the ADC PBS buffer was adjusted to pH = 9 with 1M Tris buffer, incubated at 37°C for 24 hours, and then adjusted to pH 6.5 with 1mol / L citric acid buffer to obtain conjugate V-6 (3.3mg / mL, 5.5ml). a-I Can be detected by LC-MS.

[1139] 2.8 Conjugate V-13

[1140]

[1141] At 37°C, a solution of Humira antibody in PB buffer (0.05 M PBS buffer at pH 6.5; 2.5 ml, 9.96 mg / ml, 0.168 nmol) was added with a prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol). The solution was placed in a water bath shaker and shaken at 37°C for 3 hours to stop the reaction. The reaction solution was cooled to 25°C in a water bath, diluted to 5.0 mg / ml, and 2.0 ml of the solution was removed for further reaction.

[1142] Compound IV-12 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, added to the above 2.0 mL solution, and placed in a water bath shaker. The reaction was shaken at 25°C for 3 hours to stop the reaction. The reaction solution was desalted and purified using a Sephadex G25 gel column (elution phase: 0.05 M histidine buffer solution at pH 5.5, containing 0.001 M EDTA) to obtain conjugate V-13 (3.5 mg / mL, 5.6 mL).

[1143] N a-I Can be detected by LC-MS.

[1144] 2.9 Conjugate V-12

[1145]

[1146] At 37°C, a solution of Humira antibody in PB buffer (0.05 M PBS buffer at pH 6.5; 2.5 ml, 9.96 mg / ml, 0.168 nmol) was added with a prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol). The solution was placed in a water bath shaker and shaken at 37°C for 3 hours to stop the reaction. The reaction solution was cooled to 25°C in a water bath, diluted to 5.0 mg / ml, and 2.0 ml of the solution was removed for further reaction.

[1147] Compound IV-11 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA, added to the above 2.0 mL solution, and placed in a water bath shaker. The reaction was shaken at 25°C for 3 hours, and the reaction was stopped. The reaction solution was desalted and purified using a Sephadex G25 gel column (elution phase: 0.05 M histidine buffer solution at pH 5.5, containing 0.001 M EDTA) to obtain conjugate V-12 (4.4 mg / mL, 4.3 mL).

[1148] N a-I Can be detected by LC-MS.

[1149] 2.10 Conjugate V-49

[1150]

[1151] At 37°C, a solution of Humira antibody in PB buffer (0.05 M PBS buffer at pH 6.5; 2.5 ml, 9.96 mg / ml, 0.168 nmol) was added with a prepared aqueous solution of tris(2-carboxyethyl)phosphine (10 mM, 0.054 mL, 0.54 nmol). The solution was placed in a water bath shaker and shaken at 37°C for 3 hours to stop the reaction. The reaction solution was cooled to 25°C in a water bath, diluted to 5.0 mg / ml, and 2.0 ml of the solution was removed for further reaction.

[1152] Compound IV-60 (1.67 mg, 2.02 nmol) was dissolved in 0.10 mL of DMA and added to the above 2.0 mL solution. The mixture was placed in a water bath shaker and shaken at 25°C for 3 hours to stop the reaction. The reaction solution was desalted and purified using a Sephadex G25 gel column (elution phase: 0.05 M histidine buffer solution at pH 5.5, containing 0.001 M EDTA) to obtain conjugate V-53 (2.7 mg / mL, 4.7 mL).

[1153] N a-I Can be detected by LC-MS.

[1154] 2.11 Conjugate V-70 (Humira-coujugate 5)

[1155]

[1156] To a PB bu...

Claims

1. Compounds of formula I: or a pharmaceutically acceptable salt thereof, wherein R1, R2 are each independently selected from: H, F, Cl and methyl; R3 is selected from: -CH2Cl, -CH2SH, -CH2OH, -OCH3, -OCH2F, -OCH2Cl, -OCH2CN, -OCH2CH3, -SH, -SCH2F, -SCH2Cl, -SCH2CF3 and -SCH2CN; R4 and R5 are each independently selected from the group consisting of hydrogen, protium, deuterium, F, Cl, -OH, -NH2, and C1-C6 alkyl; n is selected from 1, 2 or 3; B is W is A1 is a benzene ring; X is -O-, -S- or -NH-; Y1 is absent or selected from the group consisting of: protium, deuterium, hydroxyl, thiol, amino and C1-C6 alkyl; m is 0 or 1.

2. A compound of formula I according to claim 1 or a pharmaceutically acceptable salt thereof, selected from: in, R1, R2, R3, R4, R5, B, W, A1, X, Y1, n and m are as defined in claim 1.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein said R4 and R5 are each independently selected from: H, F, Cl, -OH, -NH2 and C1-C6 alkyl.

4. The compound according to claim 3 or a pharmaceutically acceptable salt thereof, wherein the -(CR4R5) n - are each independently selected from: -CH2-, and -CH2CH2-.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Selected from:

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is selected from in, Said X is selected from -O-, -S- and -NH-; R4 and R5 are each independently selected from the following group: hydrogen, protium, deuterium, F, Cl, -OH, -NH2 and C1-C6 alkyl; said n is selected from 1, 2 or 3; The Y1 is absent or selected from: hydroxyl, thiol, amino and C1-C6 alkyl; Said R1 and R2 are each independently selected from hydrogen, fluorine, chlorine and methyl; The R3 is selected from: -CH2Cl, -CH2SH, -CH2OH, -OCH3, -OCH2F, -OCH2Cl, -OCH2CN, -OCH2CH3, -SH, -SCH2F, -SCH2Cl, -SCH2CF3 and -SCH2CN.

7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

8. Compounds of formula IVa or IVb: or a pharmaceutically acceptable salt thereof, wherein R1, R2 are each independently selected from: H, F, Cl and methyl; R3 is selected from: -CH2Cl, -CH2SH, -CH2OH, -OCH3, -OCH2F, -OCH2Cl, -OCH2CN, -OCH2CH3, -SH, -SCH2F, -SCH2Cl, -SCH2CF3 and -SCH2CN; R4 and R5 are each independently selected from the group consisting of hydrogen, protium, deuterium, F, Cl, -OH, -NH2, and C1-C6 alkyl; The n is selected from 1, 2 or 3; B is W is Y1 is absent or selected from the group consisting of: protium, deuterium, hydroxyl, thiol, amino and C1-C6 alkyl, m is 0 or 1; X is selected from: -O-, -S- and -NH-; Y2 is selected from -O-, -S- and -NH-; Tr is absent or selected from the following structures: and (1) When L1 is coupled to the ligand via a thiol group, L1 is selected from: Among them, the R L1a , R L1b , R L1c Each is independently selected from: hydrogen, methyl, ethyl, phenyl and benzyl; (2) When L1 is coupled to the ligand via an amino group, L1 is selected from: (3) When L1 is coupled by click chemistry, L1 is selected from: Among them, the R L1a , R L1b , R L1c Each is independently selected from: hydrogen, methyl, ethyl, phenyl and benzyl; L2 is absent, or L2 contains a PEG branch or a PEG linear chain; L3 is selected from the following group: glycine-glycine-phenylalanine-glycine (GGFG), alanine-alanine-alanine-glycine (AAAG), glycine-glycine-glycine-glycine (GGGG), valine-alanine-glycine (VAG), valine-citrulline-glycine (VCG), alanine-alanine-glycine (AAG), alanine-alanine-alanine (AAA), valine-alanine (VA), valine-citrulline (VC), alanine-alanine-glycine (AAG), alanine-alanine-alanine (AAA), valine-alanine (VA), valine-citrulline (VC), alanine-alanine (AA), glutamate-alanine-glycine-glycine (EAGG), glycine-glutamate-alanine-glycine (GEAG), glycine-glutamate-glycine-glycine (GEGG), glutamate-glycine-glycine (EGG), glutamate-alanine-glycine (EAG), valine-lysine-glycine (VKG), glycine-glutamate-glycine (GEG), glutamate-alanine (EA), glutamate-glycine (EG), and glycine-glutamate (GE).

9. The compound of formula IVa or IVb or a pharmaceutically acceptable salt thereof according to claim 8, wherein said R4 and R5 are each independently selected from the following group: H, F, Cl, -OH, -NH2 and C1-C6 alkyl; and said n is selected from 1, 2 or 3.

10. The compound of formula IVa or IVb or a pharmaceutically acceptable salt thereof according to claim 8, wherein Y1 is absent or selected from the group consisting of: hydroxyl, thiol, amino and C1-C6 alkyl.

11. A compound of formula IVa or IVb according to claim 8 or a pharmaceutically acceptable salt thereof, wherein Selected from 12. The compound of formula IVa or IVb or a pharmaceutically acceptable salt thereof according to claim 8, wherein the compound of formula IVa or IVb is selected from:

13. A ligand-drug conjugate or a pharmaceutically acceptable salt thereof, wherein the ligand-drug conjugate has the following structure: in, R1, R2 are each independently selected from: H, F, Cl and methyl; R3 is selected from: -CH2Cl, -CH2SH, -CH2OH, -OCH3, -OCH2F, -OCH2Cl, -OCH2CN, -OCH2CH3, -SH, -SCH2F, -SCH2Cl, -SCH2CF3 and -SCH2CN; R4 and R5 are each independently selected from the group consisting of H, protium, deuterium, F, Cl, -OH, -NH2, and C1-C6 alkyl; The n is selected from 1, 2 or 3; B is W is Y1 is absent or selected from the group consisting of: protium, deuterium, hydroxyl, thiol, amino and C1-C6 alkyl, m is 0 or 1; X is selected from: -O-, -S- and -NH-; Y2 is selected from -O-, -S- and -NH-; Tr, L3, L2 and L1 are as defined in claim 8; Ab represents the ligand that can bind to the target, N a-I Any number from 1 to 10.

14. The ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to claim 13, wherein said R4 and R5 are each independently selected from the following group: H, F, Cl, -OH, -NH2 and C1-C6 alkyl; and said n is selected from 1, 2 or 3.

15. The ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to claim 13, wherein the Y1 is absent or selected from the group consisting of: hydroxyl, thiol, amino and C1-C6 alkyl.

16. The ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to claim 13, wherein Selected from 17. The ligand-drug conjugate according to claim 13 or a pharmaceutically acceptable salt thereof, in, Said X is selected from -O-, -S- and -NH-; R4 and R5 are each independently selected from the group consisting of H, protium, deuterium, F, Cl, -OH, -NH2, and C1-C6 alkyl; The n is selected from 1, 2 or 3; The Y1 is absent or selected from: hydroxyl, thiol, amino and C1-C6 alkyl; Said R1 and R2 are each independently selected from hydrogen, fluorine, chlorine and methyl; The R3 is selected from: -CH2Cl, -CH2SH, -CH2OH, -OCH3, -OCH2F, -OCH2Cl, -OCH2CN, -OCH2CH3, -SH, -SCH2F, -SCH2Cl, -SCH2CF3 and -SCH2CN.

18. The ligand-drug conjugate according to claim 13 or a pharmaceutically acceptable salt thereof, in, The structural unit -Tr-L3-L2-L1- is selected from:

19. The ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 13 to 18, wherein the ligand is selected from the group consisting of: anti-TNFα antibody or an antigen-binding fragment thereof, anti-CD40 antibody or an antigen-binding fragment thereof, and anti-IFNAR1 antibody or an antigen-binding fragment thereof.

20. The ligand-drug conjugate of claim 19 or a pharmaceutically acceptable salt thereof, wherein the antibody is selected from the group consisting of human antibodies, humanized antibodies, chimeric antibodies, multispecific antibodies, monoclonal antibodies and polyclonal antibodies; and the antigen-binding fragment is selected from the group consisting of Fab, Fab', F(ab')2, Fv, scFv, diabodies, Fd, dAb, VHH, large antibodies and complementarity determining region (CDR) fragments.

21. The ligand-drug conjugate according to any one of claims 13 to 18, or a pharmaceutically acceptable salt thereof, wherein the ligand is selected from the group consisting of: Adalimumab, Iscalimab (CFZ533), Anifrolumab (MEDI-546), Infliximab, Afelimomab, Golimumab, BIIB059, 8c11.

22. The ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 13 to 18, wherein the ligand-drug conjugate has the following structure: in, Ab represents the ligand that can bind to the target, N a-I Any number from 1 to 10; Said X is selected from -O-, -S- and -NH-; R4 and R5 are each independently selected from the group consisting of H, protium, deuterium, F, Cl, -OH, -NH2, and C1-C6 alkyl; The n is selected from 1, 2 or 3; The Y1 is absent or selected from: hydroxyl, thiol, amino and C1-C6 alkyl; Said R1 and R2 are each independently selected from hydrogen, fluorine, chlorine and methyl; The R3 is selected from: -CH2Cl, -CH2SH, -CH2OH, -OCH3, -OCH2F, -OCH2Cl, -OCH2CN, -OCH2CH3, -SH, -SCH2F, -SCH2Cl, -SCH2CF3 and -SCH2CN.

23. The ligand-drug conjugate or a pharmaceutically acceptable salt thereof according to any one of claims 13 to 18, wherein the ligand-drug conjugate is selected from the following structures: Where N a-I is any number from 1 to 10, and Ab is selected from an antibody or an antigen-binding fragment thereof.

24. The ligand-drug conjugates represented by the following formula or their pharmaceutically acceptable salts: Where N a-I Any number from 1 to 10.

25. A pharmaceutical composition comprising the compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof and / or the ligand-drug conjugate according to any one of claims 13 to 24, and optionally a pharmaceutically acceptable carrier.

26. Use of a compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, a ligand-drug conjugate according to any one of claims 13 to 24, and / or a pharmaceutical composition according to claim 25 in the preparation of a medicament for preventing and / or treating diseases and / or symptoms associated with glucocorticoid receptor signal transduction, wherein the diseases and / or symptoms are selected from the following group: rheumatoid arthritis, systemic lupus erythematosus, scleroderma, Sjögren's syndrome, ankylosing spondylitis, Wegener's granulomatosis and systemic sclerosis, autoimmune hemolytic anemia Blood, pernicious anemia, idiopathic thrombocytopenic purpura, idiopathic thrombocytopenia and vasculitis, multiple sclerosis, myasthenia gravis and Guillain-Barre syndrome, ulcerative colitis, Crohn's disease, autoimmune diseases and atrophic gastritis, IgA nephropathy, primary nephrotic syndrome, autoimmune glomerulonephritis, pulmonary hemorrhagic syndrome and lupus nephritis, type I diabetes, Grave's disease, Hashimoto's thyroiditis, primary adrenal cortical atrophy and chronic thyroiditis, psoriasis, herpes vulgaris, cutaneous lupus erythematosus, dermatomyositis and polymyalgia rheumatica and asthma.

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