Selective drug release from conjugates of bioactive compounds

Abstract

This disclosure relates to compounds having formula (X), or pharmaceutically acceptable salts, tautomers, stereoisomers, enantiomers, diastereomers, or mixtures thereof, and conjugates containing such compounds. S-D (X)

Description

[0001] Cross-references to related applications

[0002] This application claims priority to PCT application PCT / CN2023 / 130153, filed on November 7, 2023, and PCT application PCT / CN2024 / 125035, filed on October 15, 2024, the contents of which are incorporated herein by reference in their entirety for all purposes. Technical Field

[0003] This disclosure relates to compounds having formula (X) or pharmaceutically acceptable salts thereof, tautomers, stereoisomers, enantiomers, diastereomers, or mixtures thereof, and conjugates containing such compounds. Background Technology

[0004] This invention relates to drug conjugate compounds and compositions thereof, including antibody-drug conjugates (ADCs) and ligand-drug conjugates (LDCs), which exhibit improved selectivity for targeted cells compared to untargeted cells. The invention also relates to drugs and drug-linkers that can be used as part of drug conjugate compounds, and compositions thereof.

[0005] Traditional ADCs and LDCs exhibit bioactivity in targeted cells by binding to the target moiety of the antibody or ligand exhibiting recognition of the conjugate: the target moiety binds to the target moiety and then enters the cell through internalization of the bound conjugate. Selectivity for targeted cells compared to non-targeted cells is primarily achieved by traditional ADCs and LDCs because the target moiety is more abundant on targeted cells than on non-targeted normal cells (which are expected to be unaffected by the conjugate). Conditional release of conjugated compounds with cytotoxicity in free form is influenced by the internalization of the conjugate by intracellular proteases, followed by enzymatic processing of the peptide-based linker unit of the conjugate. By optimizing the selectivity of specific lysosomal proteases that are believed to be upregulated in cancer cells, premature release of cytotoxic compounds from traditional peptide-based ADCs and LDCs has been reduced, which would otherwise lead to undesirable side effects. Since the proteases responsible for intracellular processing in conventional ADCs and LDCs are universal across all cells, the selectivity for targeted cells is primarily due to the higher abundance of the targeted moiety on cells expected to act on the conjugate, despite differences in intracellular activity levels of the proteases processed in targeted cancer cells and non-targeted normal cells. However, this approach does not account for potential differences in exposure to the released cytotoxic compounds between tumor and normal tissues, differences that the ADCs and LDCs of this invention currently utilize.

[0006] Therefore, the peptide sequences of conventional ADCs and LDCs, designed to selectively target intracellular proteases upregulated in cancer cells within tumor tissue, can still be affected by proteases restricted to normal tissues. Such effects can occur within the microenvironment of normal tissues or within cells after immune-specific or non-specific uptake, leading to on-target or off-target toxicity, respectively. These toxicities present a more acute challenge for the targeted delivery of highly cytotoxic compounds. Therefore, it is believed that ADCs and LDCs with improved peptide sequences will improve therapeutic tolerability, reducing exposure to normal tissues and thus exposure to released cytotoxic compounds compared to conventional peptide-based ADCs and LDCs, while maintaining the efficacy provided by these conventional conjugates.

[0007] Furthermore, it is believed that ADCs and LDCs with improved peptide sequences will also reduce exposure to released cytotoxic compounds, which could help improve therapy tolerability. These ADCs and LDCs are more readily proteased by tumor tissues than by normal tissues, compared to conventional peptide-based ADCs and LDCs. Determining these differences in proteolysis using tissue homogenates should capture those differences driven by the microenvironment of these tissues and / or after internalization. Summary of the Invention

[0008] To provide a solution to this problem in the art, this paper discloses ADCs and LDCs with peptide-based linker units whose sequences enable tumor-effect targeted cells to be selectively exposed to cytotoxic compounds released from the conjugates compared to normal tissue cells exposed to free cytotoxic substances, thereby improving conjugate tolerance while retaining the efficacy of conventional peptide-based conjugates. In cancer treatment in mammalian subjects, this differential exposure may be due to the greater selectivity of ADCs and LDCs with selective peptide sequences in proteolysis within tumor tissues compared to proteolysis in normal tissues, compared to proteolysis with conventional peptide-based conjugates. Because altering the peptide sequence can also affect the physicochemical properties of the conjugate compound, greater exposure occurs due to improved biodistribution to tumor tissues rather than normal tissues and / or improved disposal once distributed to these tissues, which can preferentially retain the conjugate compound in tumor tissues and / or preferentially eliminate the conjugate compound from normal tissues, respectively. These biodistribution effects may even become the dominant factor in preferential proteolysis, which is difficult to observe in vivo.

[0009] Therefore, conjugate compounds having peptide sequences that enhance the exposure of free cytotoxic compounds to tumor tissues compared to normal tissues should exhibit reduced undesirable toxicity because the peptide sequence is less sensitive to proteolysis in normal tissues or cells compared to proteolysis in tumor tissues or their cells, and / or due to the improved pharmacokinetic properties of conjugate compounds incorporating peptide sequences that are more favorable to tumor tissues than normal tissues.

[0010] Drug release from the tumor-targeting device disclosed herein can ideally occur within the tumor microenvironment, allowing the active payload to subsequently diffuse and internalize into neighboring tumor cells. Indeed, proteolytic enzymes such as cathepsin B, urokinase-type plasminogen activator (uPA), lysyl oxidase (LOX), and matrix metalloproteinases (MMPs) are involved in cancer progression features such as angiogenesis, invasion, and metastasis.

[0011] In a first aspect, a compound having formula (X) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof is provided:

[0012] SD (X)

[0013] in,

[0014] S is a linker, and the linker contains a peptide-cleavable unit that contains a tripeptide having the sequence -P3-P2-P1-.

[0015] P3 is selected from serine, tyrosine, or their analogues;

[0016] P2 is selected from glycine, serine, or their analogues;

[0017] P1 is selected from citrulline, arginine, glutamic acid or their analogues;

[0018] D represents the drug component.

[0019] On the other hand, a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof is provided:

[0020] LPYD (I)

[0021] in,

[0022] L is represented by L1-L2-X-;

[0023] L1 is the ligand covalently bound portion and can be alternatively selected from... and ;

[0024] -L2-X- is an extended subunit;

[0025] L2 is selected from C 1-20 Alkylene, -C 2-20 imide and C 2-20 Idemynyl group, in which C 1-20 The 1, 2, 3, 4, 5, 6, 7 or 8 non-adjacent carbon atoms in the alkylene group may optionally be replaced by O or S;

[0026] X either does not exist or is -C(O)-;

[0027] L can be arbitrarily divided by 1, 2, 3, 4 or 5 Rs. L Group substitution;

[0028] R L Selected from H, D, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0029] Y is the spacer subunit;

[0030] D represents the drug component;

[0031] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0032] P3 is selected from serine or its analogues;

[0033] P2 is selected from glycine or its analogues;

[0034] P1 is selected from citrulline, arginine, glutamic acid, or their analogues.

[0035] On the other hand, this disclosure provides a coupling having formula (II):

[0036] T-(S'-D) k (II)

[0037] in,

[0038] T stands for the target region;

[0039] S' is a linker, which is a divalent group formed by linking S and T;

[0040] The linker contains a peptide-cleavable unit that contains a tripeptide having the sequence -P3-P2-P1-, where P3, P2, and P1 are as defined in the context.

[0041] D represents the drug component;

[0042] k ranges from 1 to approximately 20.

[0043] On the other hand, this disclosure provides the conjugates disclosed herein, as well as optional pharmaceutically acceptable one or more carriers or one or more excipients.

[0044] On the other hand, this disclosure provides pharmaceutical compositions comprising the conjugates disclosed herein and one or more pharmaceutically acceptable excipients, the pharmaceutical compositions further comprising one or more other therapeutic agents.

[0045] On the other hand, this disclosure provides kits comprising pharmaceutical compositions including the conjugates disclosed herein, one or more other therapeutic agents, and one or more pharmaceutically acceptable carriers, one or more adjuvants, or one or more mediators.

[0046] On the other hand, this disclosure provides for the use of a compound or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, a conjugate thereof, or a composition disclosed herein in the manufacture of a medicament for the treatment and / or prevention of disease.

[0047] On the other hand, this disclosure provides a method for preventing and / or treating a disease in a subject in need, the method comprising administering the conjugate disclosed herein to the subject.

[0048] In another aspect, the method further includes administering a second therapeutic agent to the subject, preferably wherein the second therapeutic agent is selected from antibodies, chemotherapeutic agents, and small molecule drugs.

[0049] On the other hand, this disclosure provides compounds or pharmaceutically acceptable salts thereof, enantiomers, diastereomers thereof, mixtures thereof, conjugates thereof, or compositions disclosed herein for use in the treatment and / or prevention of diseases.

[0050] In a particular embodiment, the disease is selected from cancer, infectious diseases, inflammatory diseases, autoimmune diseases, and immunodeficiency diseases.

[0051] It should be understood that one, some, or all of the features of the various embodiments described herein can be combined to form other embodiments of this disclosure. These and other aspects of this disclosure will become apparent to those skilled in the art. These and other embodiments of this disclosure will be further described in the following detailed description.

[0052] definition

[0053] The following lists definitions of various terms used to describe the invention. Unless otherwise limited individually or as part of a larger group in a particular case, these definitions apply to the terminology used throughout the specification and claims.

[0054] Chemical definition

[0055] When listing a range of values, the aim is to cover every value within that range and its subranges. For example, "C 1-6 The term "alkyl" is intended to encompass C1, C2, C3, C4, C5, C6, and C6 alkyl groups. 1-6 C 1-5 C 1-4 C 1-3 C 1-2 C 2-6 C 2-5 C 2-4 C 2-3 C 3-6 C 3-5 C 3-4 C 4-6 C 4-5 And C 5-6 alkyl.

[0056] “C 1-20 "Alkyl" refers to a free radical of a straight-chain or branched saturated hydrocarbon group having 1 to 20 carbon atoms. 1-10 "Alkyl" refers to a free radical of a straight-chain or branched saturated hydrocarbon group having 1 to 10 carbon atoms. 1-6 "alkyl" refers to a free radical of a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms. In some embodiments, C 1-4 Alkyl groups are preferred. C 1-6 Examples of alkyl groups include methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentyl (C5), pentyl (C5), neopentyl (C5), 3-methyl-2-butyl (C5), tert-pentyl (C5), and n-hexyl (C6). The term "C" is used in conjunction with alkyl groups. 1-6 "Alkyl" also includes heteroalkyl, wherein one or more (e.g., 1, 2, 3, or 4) carbon atoms are substituted with heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus). The alkyl group may optionally be substituted with one or more substituents, such as 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. Common abbreviations for alkyl include Me (-CH3), Et (-CH2CH3), iPr (-CH(CH3)2), nPr (-CH2CH2CH3), n-Bu (-CH2CH2CH2CH3), or i-Bu (CH2CH(CH3)2).

[0057] Unless otherwise stated, "C" is used alone or in combination with other terms. 1-6"Alkoxy" refers to an alkyl group having 1 to 6 carbon atoms connected to the rest of the molecule by an oxygen atom, such as methoxy, ethoxy, 1-propoxy, 2-propoxy (isopropoxy), and higher homologues and isomers. In some embodiments, C 1-4 Alkyl groups are preferred.

[0058] “C 2-20 "Alkenyl" refers to a free radical of a straight-chain or branched hydrocarbon group having 2 to 20 carbon atoms and at least one carbon-carbon double bond. 2-10 "Alkenyl" refers to a free radical of a straight-chain or branched hydrocarbon group having 2 to 10 carbon atoms and at least one carbon-carbon double bond. In some embodiments, C 2-6 Alkenyl groups are preferred. In some embodiments, C is more preferred. 2-4 Alkenyl. C 2-6 Examples of alkenyl groups include vinyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), pentenyl (C5), pentadienyl (C5), hexenyl (C6), etc. The term "C" is used in conjunction with these groups. 2-6 "Alkenyl" also includes heteroalkenyl groups, wherein one or more (e.g., 1, 2, 3, or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus). The alkenyl group may optionally be substituted by one or more substituents, such as 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0059] “C 2-20 "Alkyne" refers to a free radical of a straight-chain or branched hydrocarbon group having 2 to 20 carbon atoms, at least one carbon-carbon triple bond, and optionally one or more carbon-carbon double bonds. 2-10 "Alkyne" refers to a free radical of a straight-chain or branched hydrocarbon group having 2 to 10 carbon atoms, at least one carbon-carbon triple bond, and optionally one or more carbon-carbon double bonds. In some embodiments, C 2-6 Alkyne groups are preferred. In some embodiments, C is more preferred. 2-4 Alkynyl group. C 2-6 Examples of alkynyl groups include, but are not limited to, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), pentyynyl (C5), and hexynyl (C6). The term "C" is used in conjunction with other alkynyl groups. 2-6 "Alkyne" also includes heteroyne, in which one or more (e.g., 1, 2, 3, or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus). The alkynyl group may be substituted by one or more substituents, such as 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0060] “C 1-20"Alkylene" refers to the process of removing C 1-20 The alkyl group is a divalent group formed by the other hydrogen atom of the alkyl group, and can be a substituted or unsubstituted alkylene group. In some embodiments, C 1-10 Alkylene is preferred. In some embodiments, C 1-6 Alkylene is preferred. In some embodiments, C 1-4 Alkylenes are particularly preferred. Unsubstituted alkylene groups include, but are not limited to, methylene (-CH2-), ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), butylene (-CH2CH2CH2CH2-), pentylene (-CH2CH2CH2CH2CH2-), and hexylene (-CH2CH2CH2CH2CH2CH2-). Examples of substituted alkylene groups, such as those substituted with one or more alkyl (methyl) groups, include, but are not limited to, substituted methylene (-CH(CH3)-, -C(CH3)2-), substituted ethylene (-CH(CH3)CH2-, -CH2CH(CH3)-, -C(CH3)2CH2-, -CH2C(CH3)2-), substituted propylene (-CH(CH3)CH2CH2-, -CH2CH(CH3)CH2-, -CH2CH2CH(CH3)-, -C(CH3)2CH2CH2-, -CH2C(CH3)2CH2-, -CH2CH2C(CH3)2-), etc.

[0061] “C 2-20 "Alkenyl" refers to the group formed by removing C 2-20 The other hydrogen atom of the alkenyl group forms a divalent group, and it can be a substituted or unsubstituted alkenyl group. In some embodiments, C 2-10 Alkenyl groups are preferred. In some embodiments, C 2-6 Alkenyl groups are preferred. In some embodiments, C 2-4 Alkenyl groups are particularly preferred. Exemplary unsubstituted alkenyl groups include, but are not limited to, vinylidene (-CH=CH-) and propenylidene (e.g., -CH=CHCH2-, -CH2-CH=CH-). Exemplary substituted alkenyl groups, such as alkenyl groups substituted with one or more alkyl (methyl) groups, include, but are not limited to, substituted vinylidene (-C(CH3)=CH-, -CH=C(CH3)-), substituted propenylidene (e.g., -C(CH3)=CHCH2-, -CH=C(CH3)CH2-, -CH=CHCH(CH3)-, -CH=CHC(CH3)2-, -CH(CH3)-CH=CH-, -C(CH3)2-CH=CH-, -CH2-C(CH3)=CH-, -CH2-CH=C(CH3)-), etc.

[0062] “C 2-20 "Alynyl group" refers to the group formed by removing C 2-20 The other hydrogen atom of the alkynyl group forms a divalent group, and it can be a substituted or unsubstituted alkynyl group. In some embodiments, C 2-10 Amyynyl group is preferred. In some embodiments, C 2-6 Amyynyl group is preferred. In some embodiments, C 2-4 The alkynyl group is particularly preferred. Exemplary alkynyl groups include, but are not limited to, ethynyl (-C≡C-), substituted or unsubstituted propynyl (-C≡CCH2-), etc.

[0063] "Halogen" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I).

[0064] “C 1-6 "Halogenated alkyl" refers to the above "C" group that has been substituted with one or more halogen groups. 1-6 Alkyl group. "C" 1-6 "Haloalkoxy" refers to the above "C" group that has been substituted with one or more halogen groups. 1-6 "Alkoxy". Examples include monohalogenated, dihalogenated, and polyhalogenated alkyl groups, including perhalogenated alkyl groups. The monohalogen substituent in this group can be an iodine, bromine, chlorine, or fluorine atom; the dihalogenated and polyhalogenated substituents can be two or more identical halogen atoms or combinations of different halogens. Preferred examples of haloalkyl groups include monofluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl, dichloromethyl, trichloromethyl, pentafluoroethyl, heptafluoropropyl, difluorochloromethyl, dichlorofluoromethyl, difluoroethyl, difluoropropyl, dichloroethyl, and dichloropropyl. The haloalkyl group can be substituted at any available attachment point, for example, substituted by 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0065] “C 3-10 "Cycloalkyl" refers to a free radical having 3 to 10 ring carbon atoms and zero heteroatoms, which is a non-aromatic cyclic hydrocarbon group. In some embodiments, C 3-7 Cycloalkyl groups are particularly preferred, C 4-6 Cycloalkyl groups are preferred, and C... 3-5Cycloalkyl is even more preferred. Cycloalkyl also includes a ring system in which the cycloalkyl ring as defined above is fused with one or more aryl or heteroaryl groups, wherein the attachment point is on the cycloalkyl ring, and in this case, the number of carbons continues to represent the number of carbons in the cycloalkyl ring system. Exemplary cycloalkyl groups include, but are not limited to, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cyclohepttrienyl (C7), etc.

[0066] The terms “heterocyclic,” “heterocyclic group,” or “heterocyclic alkyl” are used interchangeably and refer to a fused, bridged, or spirocyclic system of non-aromatic rings, bicyclic, or tricyclic groups, wherein (i) each ring system contains at least one heteroatom independently selected from oxygen, sulfur, and nitrogen; (ii) each ring system may be saturated or unsaturated; (iii) the nitrogen and sulfur heteroatoms may optionally be oxidized; (iv) the nitrogen heteroatom may optionally be quaternized; (v) any of the above rings may be fused to an aromatic ring; and (vi) the remaining ring atoms are carbon atoms that may optionally be oxo-substituted or optionally substituted with exocyclic alkenyl, imine, or oxime double bonds.

[0067] "3- to 10-membered heterocyclic groups" refer to free radicals of 3- to 10-membered non-aromatic ring systems having a cyclic carbon atom and 1 to 5 cyclic heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon. In heterocyclic groups containing one or more nitrogen atoms, the attachment point can be a carbon atom or a nitrogen atom, provided the valence allows. In some embodiments, 3- to 9-membered heterocyclic groups are preferred, being free radicals of 3- to 9-membered non-aromatic ring systems having a cyclic carbon atom and 1 to 5 cyclic heteroatoms; in some embodiments, 3- to 7-membered heterocyclic groups are preferred, being free radicals of 3- to 7-membered non-aromatic ring systems having a cyclic carbon atom and 1 to 4 cyclic heteroatoms; 3- to 6-membered heterocyclic groups are preferred, being free radicals of 3- to 6-membered non-aromatic ring systems having a cyclic carbon atom and 1 to 3 cyclic heteroatoms. The free radical of the ring system; 4- to 7-membered heterocyclic groups are preferred, which are free radicals of a 4- to 7-membered non-aromatic ring system having a ring carbon atom and 1 to 3 ring heteroatoms; 4- to 6-membered heterocyclic groups are preferred, which are free radicals of a 4- to 6-membered non-aromatic ring system having a ring carbon atom and 1 to 3 ring heteroatoms; 5- to 6-membered heterocyclic groups are more preferred, which are free radicals of a 5- to 6-membered non-aromatic ring system having a ring carbon atom and 1 to 3 ring heteroatoms. The heterocyclic group also includes a ring system in which the heterocyclic ring as defined above is fused with one or more cycloalkyl groups and the attachment point is on the cycloalkyl ring; or in which the heterocyclic ring as defined above is fused with one or more aryl or heteroaryl groups and the attachment point is on the heterocyclic ring; and in this case, the number of ring members continues to represent the number of ring members in the heterocyclic ring system. Exemplary 3-membered heterocyclic groups containing one heteroatom include, but are not limited to, aziridinyl, ethylene oxide, and thiorenyl. Exemplary 4-membered heterocyclic groups containing one heteroatom include, but are not limited to, azirrobutyl, oxetyl, and thiorenyl. Exemplary 5-membered heterocyclic groups containing one heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrophenylthio, dihydrophenylthio, pyrrolyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclic groups containing two heteroatoms include, but are not limited to, dioxolanecycloyl, oxetylfuranyl, dithiofuranyl, and oxazolidinyl-2-one. Exemplary 5-membered heterocyclic groups containing three heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclic groups containing one heteroatom include, but are not limited to, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thiorenyl. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to, piperazinyl, morpholinyl, dithiaalkyl, and dioxalyl. Exemplary 6-membered heterocyclic groups containing three heteroatoms include, but are not limited to, triazineyl. Exemplary 7-membered heterocyclic groups containing one heteroatom include, but are not limited to, azirheptanyl, oxaheptanyl, and thioheptanyl.Exemplary 5-membered heterocyclic groups fused to a C6 aryl ring (also referred to herein as 5,6-bicyclic heterocycles) include, but are not limited to: dihydroindolyl, isoindolyl, dihydrobenzofuranyl, dihydrobenzothiophenyl, benzoxazolinone, etc. Exemplary 6-membered heterocyclic groups fused to a C6 aryl ring (also referred to herein as 6,6-bicyclic heterocycles) include, but are not limited to, tetrahydroquinolinyl, tetrahydroisoquinolinyl, etc.

[0068] The 3- to 10-membered heterocyclic groups also include spiroheterocyclic groups, i.e., groups in which two rings (e.g., a heterocyclic group and a carbocyclic group) share a carbon atom, wherein at least one ring is a heterocyclic group as defined above. More specifically, the spiroheterocyclic group is a spirocycle formed by two 4-membered rings, two 5-membered rings, two 6-membered rings, one 4-membered ring and one 5-membered ring, one 4-membered ring and one 6-membered ring, or one 5-membered ring and one 6-membered ring, wherein at least one ring is a 4- to 6-membered heterocyclic group as defined above. 4- to 6-membered heterocyclic groups containing one, two, or three O, N, or S heteroatoms are preferred, and 4- to 6-membered heterocyclic groups containing one N heteroatom are more preferred. Specific spiroheterocyclic groups include, but are not limited to:

[0069] , , , , , , , , , , , , , , , , , , , , , , ,and .

[0070] “C 6-10 "Aryl" refers to a free radical of a monocyclic or polycyclic (e.g., bicyclic) 4n+2 aromatic ring system (e.g., having 6 or 10 π electrons shared in a ring array) having 6-10 ring carbon atoms and zero heteroatoms. In some embodiments, the aryl group has six ring carbon atoms ("C6 aryl"; for example, phenyl). In some embodiments, the aryl group has ten ring carbon atoms ("C6 aryl"). 10"Aryl"; for example, naphthyl, such as 1-naphthyl and 2-naphthyl). Aryl also includes ring systems in which the aryl ring as defined above is fused with one or more cycloalkyl or heterocyclic groups and the attachment point is on the aryl ring, and in this case, the number of carbon atoms continues to represent the number of carbon atoms in the aryl ring system.

[0071] "5- to 10-membered heteroaryl" refers to a free radical of a 5- to 10-membered monocyclic or bicyclic 4n+2 aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms (e.g., having 6 or 10 π electrons shared in a cyclic array), wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In heteroaryl groups containing one or more nitrogen atoms, the attachment point can be a carbon atom or a nitrogen atom, where the valence allows. A heteroaryl bicyclic system may contain one or more heteroatoms in one or both rings. Heteroaryl groups further include ring systems in which the heteroaryl ring as defined above is fused with one or more cycloalkyl or heterocyclic groups and the attachment point is on the heteroaryl ring, and in this case, the number of ring members continues to represent the number of ring members in the heteroaryl ring system. In some embodiments, 5- to 6-membered heteroaryl groups are particularly preferred, which are free radicals of a 5- to 6-membered monocyclic or bicyclic 4n+2 aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms. Exemplary 5-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyrroleyl, furanyl, and phenylthioyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl (e.g., 1,2,4-oxadiazolyl), and thiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyridinyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, but are not limited to, pyridazinyl, pyrazinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetraazinyl, respectively. Exemplary 7-membered heteroaryl groups containing one heteroatom include, but are not limited to, azepinyl, oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryl groups include, but are not limited to: indolyl, isoindolyl, indazole, benzotriazolyl, benzobenzylthio, isobenzobenzylthio, benzofuranyl, benzoisofuranyl, benzoimidazolyl, benzoxazolyl, benzoisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzoisothiazolyl, benzothiadiazolyl, indolazinyl, and purinel. Exemplary 6,6-bicyclic heteroaryl groups include, but are not limited to: naphridinyl, pteridinyl, quinolinyl, isoquinolinyl, cenolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.

[0072] It should be understood that any alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heterocyclic, etc., described herein may also be divalent or polyvalent groups when used as bonds connecting two or more groups or substituents (which may be one or more identical or different atoms). Those skilled in the art can readily determine the valence of any such group from the context in which it appears.

[0073] As used herein, the term "optionally substituted" means that the mentioned group may be substituted or unsubstituted. In one embodiment, the mentioned group is optionally substituted with zero substituents, i.e., the mentioned group is unsubstituted. In another embodiment, the mentioned group is optionally substituted with one or more additional groups, individually and independently selected from the groups described herein.

[0074] The term "hydrogen" includes both hydrogen and deuterium. Additionally, the description of an atom includes other isotopes of that atom, provided the resulting compound is pharmaceutically acceptable.

[0075] Alkyl, alkoxy, alkenyl, alkynylene, alkylene, alkenylene, alkynylene, cycloalkyl, heterocyclic, aryl, and heteroaryl groups as defined herein are optionally substituted groups. Generally, the term "substituted," whether or not preceded by the term "optionally," means that at least one hydrogen atom present on a group (e.g., a carbon or nitrogen atom) is replaced by a permitted substituent (e.g., a substituent that, upon substitution, produces a stable compound, such as a compound that does not spontaneously undergo transformation (e.g., by rearrangement, cyclization, elimination, or other reactions)). Unless otherwise stated, a "substituted" group has substituents at one or more substituted positions of the group, and when more than one position in any given structure is substituted, the substituents at each position can be the same or different. The term "substituted" is considered to include substitution with all permitted substituents of an organic compound (any substituent described herein that results in the formation of a stable compound). For the purposes of this invention, a heteroatom, such as nitrogen, may have a hydrogen substituent and / or any suitable substituent as described herein, which satisfies the valence of the heteroatom and results in the formation of a stable moiety.

[0076] Exemplary substituents on carbon atoms include, but are not limited to, halogens, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, and -OR. aa -ON(R) bb )2、-N(R bb )2、-N(R bb )3 + X - -N(OR) cc )R bb -SH, -SR aa -SSR cc -C(=O)Raa 、-CO2H、-CHO、-C(OR cc )2、-CO2R aa 、-OC(=O)R aa 、-OCO2R aa 、-C(=O)N(R bb )2、-OC(=O)N(R bb )2、-NR bb C(=O)R aa 、-NR bb CO2R aa 、-NR bb C(=O)N(R bb )2、-C(=NR bb )R aa 、-C(=NR bb )OR aa 、-OC(=NR bb )R aa 、-OC(=NR bb )OR aa 、-C(=NR bb )N(R bb )2、-OC(=NR bb )N(R bb )2、-NR bb C(=NR bb )N(R bb )2、-C(=O)NR bb SO2R aa 、-NR bb SO2R aa 、-SO2N(R bb )2、-SO2R aa 、-SO2OR aa 、-OSO2R aa 、-S(=O)R aa 、-OS(=O)R aa 、-Si(R aa )3、-OSi(R aa )3、-C(=S)N(R bb )2、-C(=O)SR aa 、-C(=S)SR aa 、-SC(=S)SR aa 、-SC(=O)SR aa 、-OC(=O)SR aa 、-SC(=O)OR aa 、-SC(=O)R aa 、-P(=O)2R aa 、-OP(=O)2Raa -P(=O)(R aa )2、-OP(=O)(R aa )2、-OP(=O)(OR cc )2、-P(=O)2N(R bb )2、-OP(=O)2N(R bb )2、-P(=O)(NR bb )2、-OP(=O)(NR bb )2、-NR bb P(=O)(OR cc )2、-NR bb P(=O)(NR bb )2、-P(R cc )2、-P(R cc )3、-OP(R cc )2、-OP(R cc )3、-B(R aa 2. -B(OR) cc )2、-BR aa (OR cc ), alkyl, haloalkyl, alkenyl, ynyl, carbocyclic, heterocyclic, aryl, and heteroaryl, wherein each alkyl, alkenyl, ynyl, carbocyclic, heterocyclic, aryl, and heteroaryl is independently bounded by 0, 1, 2, 3, 4, or 5 R groups. dd Group substitution;

[0077] Or two hydrogen atom on a carbon atom are surrounded by =O, =S, =NN(R) groups. bb )2、=NNR bb C(=O)R aa =NNR bb C(=O)OR aa =NNR bb S(=O)2R aa =NR bb or =NOR cc replace;

[0078] R aa Each instance is independently selected from alkyl, haloalkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl, and heteroaryl, or two R groups. aa Groups are linked to form heterocyclic or heteroaryl rings, wherein each alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl, and heteroaryl group is independently bounded by 0, 1, 2, 3, 4, or 5 R groups. dd Group substitution;

[0079] R bb Each instance is independently selected from hydrogen, -OH, -OR aa -N(R) cc)2、-CN、-C(=O)R aa -C(=O)N(R) cc )2、-CO2R aa -SO2R aa -C(=NR) cc OR aa -C(=NR) cc )N(R cc )2、-SO2N(R cc )2、-SO2R cc -SO2OR cc -SOR aa -C(=S)N(R) cc )2、-C(=O)SR cc -C(=S)SR cc -P(=O)2R aa -P(=O)(R aa )2、-P(=O)2N(R cc )2、-P(=O)(NR cc 2. Alkyl, haloalkyl, alkenyl, ynyl, carbocyclic, heterocyclic, aryl, and heteroaryl, or two R bb Groups are linked to form heterocyclic or heteroaryl rings, wherein each alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl, and heteroaryl group is independently bounded by 0, 1, 2, 3, 4, or 5 R groups. dd Group substitution;

[0080] R cc Each instance is independently selected from hydrogen, alkyl, haloalkyl, alkenyl, ynyl, carbocyclic, heterocyclic, aryl, and heteroaryl, or two R groups. cc Groups are linked to form heterocyclic or heteroaryl rings, wherein each alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl, and heteroaryl group is independently bounded by 0, 1, 2, 3, 4, or 5 R groups. dd Group substitution;

[0081] R dd Each instance is independently selected from halogens, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OR ee -ON(R) ff )2、-N(R ff )2、-N(R ff )3 + X - -N(OR) ee )R ff -SH, -SR ee -SSR ee -C(=O)R ee-CO2H, -CO2R ee -OC(=O)R ee -OCO2R ee -C(=O)N(R) ff )2、-OC(=O)N(R ff )2、-NR ff C(=O)R ee -NR ff CO2R ee -NR ff C(=O)N(R ff )2、-C(=NR ff OR ee -OC(=NR) ff )R ee -OC(=NR) ff OR ee -C(=NR) ff )N(R ff )2、-OC(=NR ff )N(R ff )2、-NR ff C(=NR ff )N(R ff )2、-NR ff SO2R ee -SO2N(R) ff )2、-SO2R ee -SO2OR ee -OSO2R ee -S(=O)R ee 、-Si(R ee 3. -OSi(R) ee 3. -C(=S)N(R) ff )2、-C(=O)SR ee -C(=S)SR ee -SC(=S)SR ee -P(=O)2R ee -P(=O)(R ee )2、-OP(=O)(R ee )2、-OP(=O)(OR ee 2. Alkyl, haloalkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl, and heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl, and heteroaryl group is independently bounded by 0, 1, 2, 3, 4, or 5 R groups. gg Group substitution, or two geminal R groups dd Substituents can connect to form =O or =S;

[0082] Ree Each instance is independently selected from alkyl, haloalkyl, alkenyl, alkynyl, carbocyclic, aryl, heterocyclic, and heteroaryl groups, wherein each alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl, and heteroaryl group is independently represented by 0, 1, 2, 3, 4, or 5 R groups. gg Group substitution;

[0083] R ff Each instance is independently selected from hydrogen, alkyl, haloalkyl, alkenyl, ynyl, carbocyclic, heterocyclic, aryl, and heteroaryl, or two R groups. ff Groups are linked to form heterocyclic or heteroaryl rings, wherein each alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl, and heteroaryl group is independently bounded by 0, 1, 2, 3, 4, or 5 R groups. gg Group substitution; and

[0084] R gg Each instance is independently a halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OC 1-6 Alkyl, -ON(C) 1-6 Alkyl)2, -N(C 1-6 Alkyl)2, -N(C 1-6 Alkyl)3 + X - -NH(C 1-6 Alkyl)2 + X - -NH2(C 1-6 alkyl) + X - -NH3 + X - -N(OC) 1-6 Alkyl)(C 1-6 Alkyl), -N(OH)(C 1-6 Alkyl groups, -NH(OH), -SH, -SC 1-6 Alkyl, -SS(C 1-6 Alkyl), -C(=O)(C 1-6 Alkyl group, -CO2H, -CO2(C 1-6 Alkyl), -OC (=O)(C 1-6 Alkyl), -OCO2(C 1-6 Alkyl groups, -C(=O)NH2, -C(=O)N(C 1-6 Alkyl)2、-OC(=O)NH(C 1-6 Alkyl), -NHC(=O)(C 1-6 alkyl), -N(C) 1-6 Alkyl)C(=O)(C 1-6 alkyl), -NHCO2(C 1-6Alkyl), -NHC(=O)N(C 1-6 alkyl)2、-NHC(=O)NH(C 1-6 Alkyl groups, -NHC(=O)NH2, -C(=NH)O(C 1-6 Alkyl group), -OC (=NH)(C 1-6 Alkyl group), -OC (=NH)OC 1-6 Alkyl group, -C(=NH)N(C 1-6 Alkyl)2、-C(=NH)NH(C 1-6 Alkyl groups, -C(=NH)NH2, -OC(=NH)N(C 1-6 Alkyl)2、-OC(NH)NH(C 1-6 Alkyl groups, -OC(NH)NH2, -NHC(NH)N(C 1-6 Alkyl)2, -NHC(=NH)NH2, -NHSO2(C 1-6 alkyl), -SO2N(C 1-6 alkyl)2、-SO2NH(C 1-6 Alkyl groups, -SO2NH2, -SO2C 1-6 Alkyl, -SO2OC 1-6 Alkyl, -OSO2C 1-6 Alkyl, -SOC 1-6 Alkyl, -Si(C) 1-6 Alkyl)3、-OSi(C 1-6 Alkyl)3-C(=S)N(C 1-6 Alkyl)2、C(=S)NH(C 1-6 Alkyl), C(=S)NH2, -C(=O)S(C 1-6 Alkyl), -C(=S)SC 1-6 Alkyl, -SC(=S)SC 1-6 Alkyl group, -P(=O)2(C 1-6 Alkyl), -P(=O)(C 1-6 Alkyl)2、-OP(=O)(C 1-6 Alkyl)2、-OP(=O)(OC 1-6 Alkyl)2, C 1-6 Alkyl, C 1-6 Haloalkyl, C 2- C6 alkenyl, C 2- C6 ynyl group, C 3- C7 carbon cyclogroup, C 6- C 10 Aryl, C3-C7 heterocyclic, C5-C 10 heteroaryl; or two ethryl groups gg Substituents can be linked to form =O or =S; where X - It is a counter ion.

[0085] Exemplary substituents on the nitrogen atom include, but are not limited to, hydrogen, -OH, and -OR. aa -N(R) cc )2、-CN、-C(=O)R aa -C(=O)N(R) cc )2、-CO2R aa -SO2R aa -C(=NR) bb )R aa -C(=NR) cc OR aa -C(=NR) cc )N(R cc )2、-SO2N(R cc )2、-SO2R cc -SO2OR cc -SOR aa -C(=S)N(R) cc )2、-C(=O)SR cc -C(=S)SR cc -P(=O)2R aa -P(=O)(R aa )2、-P(=O)2N(R cc )2、-P(=O)(NR cc 2. Alkyl, haloalkyl, alkenyl, ynyl, carbocyclic, heterocyclic, aryl, and heteroaryl, or two R atoms attached to a nitrogen atom. cc Groups are linked to form heterocyclic or heteroaryl rings, wherein each alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl, and heteroaryl group is independently bounded by 0, 1, 2, 3, 4, or 5 R groups. dd Group substitution, and wherein R aa R bb R cc and R dd As defined above.

[0086] Other definitions

[0087] As used herein, “cancer” means any disease caused or resulting from an inappropriately high level of cell division, an inappropriately low level of cell apoptosis, or both. Examples of cancer include, but are not limited to, leukemia (e.g., acute leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, chronic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia), polycythemia vera, lymphoma (Hodgkin's disease, non-Hodgkin's disease), Waldenström macroglobulinemia, heavy chain disease, and solid tumors.

[0088] Unless otherwise stated, the term "treatment" as used herein includes effects on a subject suffering from a particular disease, disorder, or condition that reduce the severity of the disease, disorder, or condition, or delay or slow its progression ("therapeutic treatment"). The term also includes effects that occur before a subject begins to suffer from a particular disease, disorder, or condition ("preventive treatment").

[0089] As used herein, the term “treating” refers to reversing, alleviating, inhibiting, or preventing the progression of the following: the disorder or condition to which the term applies, or one or more symptoms of such disorder or condition. The term “treating” as used herein refers to the action of treatment, which is a verb, and the latter is as defined above.

[0090] The terms “disease,” “disorder,” and “symptom” can be used interchangeably in this article.

[0091] Generally, an "effective amount" of a compound refers to an amount sufficient to elicit a target biological response. As those skilled in the art will understand, the effective amount of the compounds disclosed herein can vary depending on factors such as the desired biological endpoint, the pharmacokinetics of the compound, the disease being treated, the route of administration, and the age, health status, and symptoms of the subject. Effective amounts include therapeutically effective amounts and prophylacticly effective amounts.

[0092] Unless otherwise stated, the term "therapeuticly effective amount" as used herein refers to an amount sufficient to provide therapeutic benefit in the treatment of a disease, disorder, or condition, or to delay or minimize one or more symptoms associated with the disease, disorder, or condition. A therapeutically effective amount of a compound refers to the amount by which a therapeutic agent, when used alone or in combination with other therapies, provides therapeutic benefit in the treatment of a disease, disorder, or condition. The term "therapeuticly effective amount" may include amounts that improve overall treatment, reduce or prevent symptoms or causes of a disease or condition, or enhance the therapeutic effects of other therapeutic agents.

[0093] The term "combination" and related terms refer to the simultaneous or sequential administration of the disclosed compounds with other therapeutic agents. For example, the disclosed compounds may be administered simultaneously or sequentially with other therapeutic agents at separate unit doses, or simultaneously with other therapeutic agents at a single unit dose.

[0094] As used herein, the term “pharmaceutically acceptable” means that, within reasonable medical judgment, without undue toxicity, irritation, allergic response, etc., the substance is suitable for contact with a patient’s tissues and has a reasonable benefit / risk ratio and is effective for its intended use, including (where possible) the zwitterionic form of the compounds disclosed herein.

[0095] The term "salt" refers to the relatively non-toxic inorganic and organic acid addition salts of the compounds disclosed herein. These salts can be prepared in situ during the final separation and purification of the compounds, or by separating the salts produced by reacting the purified compounds in free base form with suitable organic or inorganic acids, respectively.

[0096] Pharmaceutically acceptable base addition salts are formed from metals or amines (e.g., alkali metal and alkaline earth metal hydroxides or organic amines). Examples of metals used as cations include sodium, potassium, magnesium, calcium, etc. Examples of suitable amines are N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucosamine, and procaine.

[0097] Salts can be prepared from inorganic acids, including sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, chlorides, bromides, and iodides. Examples of acids include hydrochloric acid, nitric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, and phosphoric acid. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, toluenesulfonate, citrate, maleate, fumarate, succinate, tartrate, naphthalate, methanesulfonate, glucono-p-ethyl, lactobionate, lauryl sulfonate, and hydroxyethyl sulfonate. Salts can also be prepared from organic acids, including aliphatic monocarboxylic and dicarboxylic acids, phenyl-substituted alkanes, hydroxyalkanes, alkanedioides, aromatic acids, aliphatic sulfonic acids, and aromatic sulfonic acids. Representative salts include acetates, propionates, octanoates, isobutyrates, oxalates, malonates, succinates, octanoates, sebacic acids, fumarates, maleates, mandelates, benzoates, chlorobenzoates, methyl benzoate, dinitrobenzoates, naphthates, benzenesulfonates, toluenesulfonates, phenylacetates, citrates, lactates, maleates, tartrates, and methanesulfonates. Pharmaceutically acceptable salts may include cations based on alkali metals and alkaline earth metals (e.g., sodium, lithium, potassium, calcium, magnesium, etc.) as well as non-toxic ammonium, quaternary ammonium, and amine cations (including but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, etc.). This also includes salts of amino acids, such as arginine salts, gluconates, galacturons, etc. (see, for example, Berge S. M. et al., “Pharmaceutical Salts,” J. Pharm., 1977;66: 1-19, which is incorporated herein by reference).

[0098] As used herein, the term "pharmaceutically acceptable salt" means that salts suitable for contact with tissues of humans and lower animals to the extent of reasonable medical judgment without undue toxicity, irritation, allergic reactions, etc., and in proportion to a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. described pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977). Salts can be prepared in situ during the final isolation and purification of the compounds of the present invention, or separately by reacting a free base functional group with a suitable organic acid. Examples of pharmaceutically acceptable salts include, but are not limited to, salts with amino groups formed from inorganic acids (e.g., hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid) or organic acids (e.g., acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid), or salts with amino groups formed by using other methods used in the art (e.g., ion exchange). Other pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogen sulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, disglucuronate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucono-heptahydrate, glyceryl phosphate, gluconate, hemisulfate, heptahydrate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, dihydroxynaphthalate, pectate, persulfate, 3-phenylpropionate, phosphate, picrate, neopentanoate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium. When appropriate, other pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyl groups having 1 to 6 carbon atoms, sulfonates, and aryl sulfonates.

[0099] In some embodiments, the compound of each formula herein is defined as including an isotopically labeled compound. An "isotopically labeled compound" is a compound in which at least one atomic position is enriched with respect to a specific isotope of a specified element to a level significantly greater than the natural abundance of that isotope. For example, one or more hydrogen atom positions in a compound may be enriched with deuterium to a level significantly greater than the natural abundance of deuterium, such as enrichment to at least 1%, preferably at least 20%, or at least 50%. For example, such deuterated compounds may be metabolized more slowly than their non-deuterated analogues, and thus exhibit a longer half-life when administered to a subject. Such compounds can be synthesized using methods known in the art, such as by employing deuterated starting materials. Unless otherwise stated, isotopically labeled compounds are pharmaceutically acceptable.

[0100] As used herein, the term "hydroxyl protecting group" refers to an unstable chemical moiety known in the art that protects the hydroxyl group from undesirable reactions during the synthetic procedure. The hydroxyl protecting group described herein may be selectively removed after one or more of the synthetic procedures. Hydroxyl protecting groups known in the art are generally described in TH Greene and PGM Wuts, Protective Groups in Organic Synthesis [, 3rd edition, John Wiley & Sons, New York (1999). Examples of hydroxyl protecting groups include benzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, tert-butoxy-carbonyl, isopropoxycarbonyl, diphenylmethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, tert-butyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, allyl, benzyl, triphenyl-methyl (triphenylmethyl), methoxymethyl, methylthiomethyl, benzyloxymethyl, 2-(trimethylsilyl)-ethoxymethyl, methanesulfonyl, trimethylsilyl, triisopropylsilyl, etc.

[0101] As used herein, the term "amino protecting group" refers to an unstable chemical moiety known in the art that protects an amino group from undesirable reactions during the synthetic procedure. Following one or more of the synthetic procedures, the amino protecting group described herein may be selectively removed. Amino protecting groups known in the art are generally described in TH Greene and PGM Wuts, Protective Groups in Organic Synthesis [, 3rd edition, John Wiley & Sons, New York (1999). Examples of amino protecting groups include, but are not limited to, methoxycarbonyl, tert-butoxycarbonyl, 9-fluorenyl-methoxycarbonyl, benzyloxycarbonyl, etc.]

[0102] The term "amino acid" refers to naturally occurring and synthetic α, β, γ, or δ amino acids, and includes, but is not limited to, amino acids found in proteins or intermediates of amino acid or protein metabolism, namely, glycine, alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, proline, serine, threonine, cysteine, tyrosine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, citrulline, arginine, and histidine. In some embodiments, the amino acid is in the L-configuration. In some embodiments, the amino acid is in the D-configuration. In some embodiments, amino acids are provided as substituents in the compounds described herein, wherein the amino acids are residues selected from the group consisting of: alanyl, valine, leucyl, isoleucyl, prolyl, phenylalanyl, tryptophanyl, methionineyl, glycyl, serinel, threonyl, cysteyl, tyrosineyl, asparagineyl, glutamineyl, asparagineyl, glutaryl, lysineyl, arginineyl, and others. Aminoacyl, β-alanyl, β-valine, β-leucyl, β-isoleucyl, β-prolyl, β-phenylalanyl, β-tryptophanyl, β-methionyl, β-glycyl, β-seryl, β-threonyl, β-cysteyl, β-tyrosyl, β-asparagineyl, β-glutamineyl, β-asparagine, β-glutamate, β-lysine, β-arginineyl, and β-histyl.

[0103] The combinations of substituents and variables contemplated by this invention are only those combinations that result in the formation of stable compounds. As used herein, the term "stable" means a compound that has sufficient stability to allow for manufacture and maintains the integrity of the compound for a sufficient period of time for use for the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

[0104] The synthesized compounds can be isolated from the reaction mixture and further purified by methods such as column chromatography, high-performance liquid chromatography, or recrystallization. Other methods for synthesizing compounds having the formula described herein will be apparent to those skilled in the art, as will be understood by those skilled in the art. Furthermore, various synthetic steps can be performed in an alternating sequence or order to give the desired compound. Synthetic chemical transformations and protecting group methodologies (protection and deprotection) used to synthesize the compounds described herein are known in the art and include, for example, those described in the following literature: R. Larock, Comprehensive Organic Transformations , 2nd edition Wiley-VCH [Wiley-VCH Publishers] (1999); TW Greene and PGM Wuts, Protective Groups in Organic Synthesis [ Protecting groups in organic synthesis], 3rd edition, John Wiley and Sons (1999); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis [Fieser and Fieser's Organic Synthesis Reagents] John Wiley and Sons (1994); and L. Paquette, editor. Encyclopedia of Reagents for Organic Synthesis [An Encyclopedia of Organic Synthesis Reagents] John Wiley and Sons (1995) and its subsequent editions.

[0105] The compounds of the present invention can be modified to enhance selective biological properties by adding appropriate functionalities. Such modifications are known in the art and may include those that increase biopermeability to a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral bioavailability, increase solubility to allow administration by injection, alter metabolism, and change the rate of excretion.

[0106] The compounds described herein contain one or more asymmetric centers, thus yielding enantiomers, diastereomers, and other stereoisomers, which can be defined according to absolute stereochemistry as (R)- or (S)- or mixtures thereof, or as (D)- or (L)- or mixtures thereof of amino acids. This invention aims to include all such possible isomers, as well as their racemic and optically pure forms. Optical isomers can be prepared from their respective optically active precursors by the procedures described above or by resolving racemic mixtures. Resolution can be performed in the presence of a resolving agent by chromatography or by repeated crystallization or by some combination of these techniques known to those skilled in the art. For further details on resolution, see Jacques, et al. Enantiomers, Racemates, and Resolutions Racemate and Split (John Willie & Sons, 1981). When the compounds described herein contain an alkene double bond, other unsaturation, or other geometrically asymmetric centers, and unless otherwise stated, the compounds are intended to include E and Z geometric isomers or cis and trans isomers. Likewise, all tautomers are intended to be included. Tautomers can be cyclic or acyclic. Any configuration of carbon-carbon double bonds appearing herein is chosen for convenience only and is not intended to specify a particular configuration unless stated herein; therefore, any carbon-carbon double bond or carbon-heteroatom double bond arbitrarily described herein as trans can be cis, trans, or a mixture of both in any proportion.

[0107] Some compounds of the present invention may also exist in separable, different stable conformations. Torsional asymmetry due to restricted rotation around asymmetric single bonds (e.g., due to steric hindrance or ring strain) may allow for the separation of different conformations. The present invention includes each conformational isomer of these compounds and mixtures thereof.

[0108] abbreviation

[0109] The abbreviations that may be used in the scheme description and the following examples are: Ac represents acetyl; AcOH represents acetic acid; AIBN represents azobisisobutyronitrile; aq. represents aqueous; BINAP represents 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl; Boc2O represents di-tert-butyl dicarbonate; Boc represents tert-butoxycarbonyl; Bpoc represents 1-methyl-1-(4-biphenyl)ethylcarbonyl; Bz represents benzoyl; Bn represents benzyl; BocNHOH represents tert-butyl N-hydroxycarbamate; t-BuOK represents potassium tert-butoxide; Bu3SnH represents tributyltin hydride; BO P represents (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate; brine represents sodium chloride aqueous solution; BSA represents N,O-bis-(trimethylsilyl)acetamide; Bz represents benzoyl; CDI represents carbonyl diimidazole; CH2Cl2 represents dichloromethane; CH3 represents methyl; CH3CN represents acetonitrile; CO2 represents carbon dioxide; COMU represents (1-cyano-2-ethoxy-2-oxoethyleneaminooxy)dimethylamino-morpholino-carbomonium hexafluorophosphate; Cs2CO3 represents cesium carbonate; CuCl represents copper chloride (I); CuI represents copper iodide ( I); dba represents dibenzylacetone; dppb represents diphenylphosphine-phosphonobutane; DBU represents 1,8-diazabicyclo[5.4.0]-undecyl-7-ene; DCC represents N,N'-dicyclohexyl-carbodiimide; DCDMH represents 1,3-dichloro-5,5-dimethylhydantoin; DCE represents 1,1-dichloroethane; DEAD represents diethyl azodicarboxylate; DIAD represents diisopropyl azodicarboxylate; DIPEA (DIEA) or (i-Pr)2EtN represents N,N,-diisopropylethylamine; DIS-Martin periodoylane represents 1,1,1-tris(acetoxy) -1,1-dihydro-1,2-benziodoyl-3-(1H)-one; DMAP represents 4-dimethylamino-pyridine; DME represents 1,2-dimethoxyethane; DMF represents N,N-dimethylformamide; DMSO represents dimethyl sulfoxide; DMT represents di(p-methoxyphenyl)phenylmethyl or dimethoxytriphenylmethyl; DMTMM represents 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholin-4-onium chloride; DPPA represents diphenylphosphoazide; EDC represents N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide; EDC HCl represents N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride; EDTA represents ethylenediaminetetraacetic acid; EtOAc represents ethyl acetate; EtOH represents ethanol; Et2O represents diethyl ether; Fmoc represents 9-fluorenylmethoxycarbonyl; HATU represents O-(7-azabenzotriazol-1-yl)-N,N,N',N',-tetramethylureonium hexafluorophosphate; HCl represents hydrogen chloride.HOBT (HOBt) represents 1-hydroxybenzotriazole; hr represents hours; K2CO3 represents potassium carbonate; K4[Fe(CN)6] represents tetrapotassium hexacyanoferrate trihydrate; n-BuLi represents n-butyllithium; i-BuLi represents isobutyllithium; t-BuLi represents tert-butyllithium; PhLi represents phenyllithium; LDA represents diisopropylaminolithium; LiTMP represents 2,2,6,6-tetramethylpiperidinium; MeOH represents methanol; Mg represents magnesium; min represents minutes; MOM represents methoxymethyl; Ms represents methanesulfonyl or -SO2-CH3; Ms2O represents methanesulfonic anhydride or methanesulfonyl anhydride; MTBE represents tert-butyl methyl ether; NaN(TMS)2 represents sodium bis(trimethylsilyl)amide; NaCl represents sodium chloride; NaH represents sodium hydride; NaHCO3 represents sodium bicarbonate or sodium bicarbonate. Na₂CO₃ represents sodium carbonate; NaOH represents sodium hydroxide; Na₂SO₄ represents sodium sulfate; NaHSO₃ represents sodium bisulfite. sulfite); Na2S2O3 represents sodium thiosulfate; NBS represents N-bromosuccinimide; NH2NH2 represents hydrazine; NH4HCO3 represents ammonium bicarbonate; NH4Cl represents ammonium chloride; NMO represents N-methylmorpholine N-oxide; NaIO4 represents sodium periodate; Ni represents nickel; NSFI represents N-fluorobenzenesulfonylimide; OH represents hydroxyl; o / n represents overnight; OsO4 represents osmium tetroxide; PE represents petroleum ether; PTSA represents p-toluenesulfonic acid; PPTS represents pyridinium p-toluenesulfonic acid; TBAF represents tetrabutylammonium fluoride; TBDMS (TBS) represents tert-butyldimethylsilyl; TBDPS represents tert-butyldiphenylsilyl; TCFH represents N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate; TEA or Et3N represents triethylamine; TES represents triethylsilyl; TESCl represents triethylsilyl chloride; TESOTf represents triethylsilyl trifluoromethanesulfonate; TFA represents trifluoroacetic acid; THF represents tetrahydrofuran; TMEDA represents N,N,N',N'-tetramethylethylenediamine; TPP or PPh3 represents triphenylphosphine; Troc represents 2,2,2-trichloroethylcarbonyl; Ts represents toluenesulfonyl or –SO2-C6H4CH3; Ts2O represents toluenesulfonic anhydride or toluenesulfonyl anhydride; TsOH represents p-toluenesulfonic acid; Pd represents palladium; Ph represents phenyl; PdOH represents palladium hydroxide; POPd represents dichlorobis(di-tert-butylphosphine-P)palladium ester (II); Pd2(dba)3 represents tris(dibenzylideneacetone)dipalladium (O); Pd(PPh3)4 represents tetra(triphenylphosphine)palladium (O);PdCl2(PPh3)2 represents trans-dichlorobis(triphenylphosphine)palladium(II); Pt represents platinum; Rh represents rhodium; rt represents room temperature; Ru represents ruthenium; satd. represents saturation; SFC represents supercritical fluid chromatography; TBS represents tert-butyldimethylsilyl; TMS represents trimethylsilyl; or TMSCl represents trimethylsilyl chloride; Zn represents zinc (powder); Zn(CN)2 represents zinc cyanide.

[0110] “PG” is an abbreviation for protecting groups, such as hydroxyl protecting groups or amino protecting groups, as defined above.

[0111] The abbreviation "PABC" refers to a self-destructing spacer. .

[0112] The abbreviation "MC" refers to the extended maleimide hexanoyl group: .

[0113] The abbreviation "MsP" refers to the extension child. . Attached Figure Description

[0114] Figure 1 The protein blots show the detection of the enzyme uPA in A431 tumor leachate and homogenate.

[0115] Figure 2 The tripeptide-based ADC was shown to have cytotoxic effects on Ag+ cancer cells (with / without uPA enzyme).

[0116] Figure 3 The tripeptide-based ADC was demonstrated to exhibit cytotoxicity against Ag-cancer cells (with / without uPA enzyme).

[0117] Figure 4 Human plasma stability of the following ADCs was demonstrated: a) RP007T2-LP13; b) RP007T2-LP17; and c) RP007T2-MC-GGFG-DXd.

[0118] Figure 5 The pharmacokinetics of the ADCs ADC1 (RP007T2-LP13) and ADC2 (RP007T2-LP17) are shown.

[0119] Figure 6 The following data show the changes in tumor volume and body weight in an A431 cell mouse tumor model following administration of ADCs of RP007T2-LP13, RP007T2-LP17, or RP007T2-MC-GGFG-DXd.

[0120] Figure 7Human plasma stability of the following ADCs was demonstrated: a) RP007T2-LP35; b) RP007T2-LP36; and c) RP007T2-MC-GGFG-DXd.

[0121] Figure 8 The pharmacokinetics of the ADCs for RP007T2-LP35 and RP007T2-LP36 are shown.

[0122] Figure 9 The following data show the changes in tumor volume and body weight in an A431 cell mouse tumor model following administration of ADCs to RP007T2-LP35, RP007T2-LP36, and RP007T2-MC-GGFG-DXd.

[0123] Figure 10 The changes in body weight of female and male BALB / c mice after administration of ADCs of RP007T2-LP35 or RP007T2-LP36 are shown.

[0124] Figure 11 Hematological test results of female and male BALB / c mice after administration of ADCs of RP007T2-LP35 or RP007T2-LP36 are shown. Detailed Implementation

[0125] As used herein, the term “compound disclosed herein” means a compound having formula (I), or a pharmaceutically acceptable salt, enantiomer, diastereomer, or mixture thereof.

[0126] In this disclosure, compounds are generally named using standard nomenclature. For compounds with asymmetric centers, it should be understood that, unless otherwise stated, all optical isomers and mixtures thereof are included. Furthermore, unless otherwise stated, all isomers and carbon-carbon double bonds included in this disclosure may exist in Z and E forms. Compounds existing in different tautomer forms, one of which is not limited to any particular tautomer, are intended to cover all tautomer forms.

[0127] I. Compounds

[0128] In one embodiment, this disclosure relates to a compound having formula (X) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof:

[0129] SD (X)

[0130] in,

[0131] S is a linker, and the linker contains a peptide-cleavable unit that contains a tripeptide having the sequence -P3-P2-P1-.

[0132] P3 is selected from serine, tyrosine, or their analogues;

[0133] P2 is selected from glycine, serine, or their analogues;

[0134] P1 is selected from citrulline, arginine, glutamic acid or their analogues;

[0135] D represents the drug component.

[0136] In another embodiment, this disclosure relates to compounds having formula (I) or pharmaceutically acceptable salts thereof, enantiomers, diastereomers, or mixtures thereof:

[0137] LPYD (I)

[0138] in,

[0139] L is represented by L1-L2-X-;

[0140] L1 is the ligand covalently bound portion and can be alternatively selected from... and ;

[0141] -L2-X- is an extended subunit;

[0142] L2 is selected from C 1-20 Alkylene, -C 2-20 imide and C 2-20 Idemynyl group, in which C 1-20 The 1, 2, 3, 4, 5, 6, 7 or 8 non-adjacent carbon atoms in the alkylene group may optionally be replaced by O or S;

[0143] X either does not exist or is -C(O)-;

[0144] L can be arbitrarily divided by 1, 2, 3, 4 or 5 Rs. L Group substitution;

[0145] R L Selected from H, D, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0146] Y is the spacer subunit;

[0147] D represents the drug component;

[0148] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0149] P3 is selected from serine or its analogues;

[0150] P2 is selected from glycine or its analogues;

[0151] P1 is selected from citrulline, arginine, glutamic acid, or their analogues.

[0152] 1.1 Variables

[0153] S

[0154] In a particular embodiment, S is a linker, and the linker comprises a peptide-cleavable unit comprising a tripeptide having the sequence -P3-P2-P1-.

[0155] In a particular embodiment, the connector S further includes a cuttable connector or a non-cuttable connector.

[0156] In a particular embodiment, the cleavable linker includes an acid-instable linker, a hydrophilic linker, a protease-sensitive linker, a light-instable linker, a hydrazone linker, a dimethyl linker, or a disulfide-containing linker.

[0157] L

[0158] In a particular embodiment, L is represented by L1-L2-X-.

[0159] In a particular embodiment, L is , where m is 1, 2, 3, 4, 5, 6, 7 or 8; alternatively, m is selected from 4, 5 or 6, and alternatively, 5.

[0160] In a particular embodiment, L is , where m is 1, 2, 3, 4, 5, 6, 7 or 8; alternatively, m is selected from 3, 4, 5 or 6, and alternatively, 4.

[0161] In a particular embodiment, L is , where n is 1, 2, 3, 4, 5, 6, 7 or 8; alternatively, n is selected from 2, 3 or 4, and alternatively, 3.

[0162] In one particular embodiment, L is not replaced; in another particular embodiment, L is replaced by one R. L Group substitution; in another particular embodiment, L is replaced by 2 R groups. L Group substitution; in another particular embodiment, L is replaced by 3 R groups. L Group substitution; in another particular embodiment, L is replaced by 4 R groups. L Group substitution; in another particular embodiment, L is replaced by 5 R groups. L Group substitution.

[0163] In a particular embodiment, RL It is H; in another particular embodiment, R L It is D; in another particular embodiment, R L It is C 1-6 Alkyl; in another particular embodiment, R L It is C 1-4 Alkyl; in another particular embodiment, R L It is C 1-6 Halogenated alkyl; in another particular embodiment, R L It is C 1-4 Halogenated alkyl groups.

[0164] L1

[0165] In a particular embodiment, L1 is a ligand covalently bound portion; in a particular embodiment, L1 is capable of forming a covalent bond with a functional group of an amino acid; in a particular embodiment, L1 is capable of forming an amide bond or a thioether bond with a functional group of an amino acid; in another particular embodiment, L1 is In another particular embodiment, L1 is .

[0166] L2 and X

[0167] In a particular embodiment, -L2-X- is an extended subunit.

[0168] In a particular embodiment, L2 is selected from C. 1-20 Alkylene, -C 2-20 imide and C 2-20 Idemynyl group, in which C 1-20 The 1, 2, 3, 4, 5, 6, 7, or 8 non-adjacent carbon atoms in the alkylene group may optionally be replaced by O or S; in another particular embodiment, L2 is C 1-10 Alkylene; in another particular embodiment, L2 is -C 0-4 Alkylene-(C 1-4 alkylene-O) 1-10 -C 0-4 Alkylene; in another particular embodiment, L2 is -C 2-10 Alkenyl; in another particular embodiment, L2 is C 2-10 Idemynyl; in another particular embodiment, L2 is -C 0-4 Alkylene-(OC) 1-4 Alkylene) 1-10 -C 0-4 Alkylene; in another particular embodiment, L2 is -(CH2). 0-4 -(CH2CH2-O) 1-10 -(CH2) 0-4 -; In another particular embodiment, L2 is -(CH2).0-4 -(O-CH2CH2) 1-10 -(CH2) 0-4 -

[0169] In a particular embodiment, L2 is C 1-10 Alkylene, wherein C 1-10 The 1, 2, 3, 4, or 5 non-adjacent carbon atoms in the alkylene group may optionally be replaced by O or S; in another particular embodiment, L2 is -(C 1-4 alkylene-O) 1-6 -; In another particular embodiment, L2 is -(OC 1-4 Alkylene) 1-6 -; In another particular embodiment, L2 is -(CH2). 0-4 -(CH2CH2-O) 1-6 -(CH2) 0-4 -; In another particular embodiment, L2 is -(CH2). 0-4 -(O-CH2CH2) 1-6 -(CH2) 0-4 -; In another particular embodiment, L2 is -C 2-10 Alkenyl; in another particular embodiment, L2 is C 2-10 Alynyl group.

[0170] In one particular embodiment, X is absent; in another particular embodiment, X is -C(O)-.

[0171] P

[0172] In a particular embodiment, P is a peptide cleavable unit, wherein the peptide cleavable unit comprises a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y.

[0173] P3

[0174] In a particular embodiment, P3 is selected from serine, tyrosine, or analogues thereof.

[0175] In a particular embodiment, P3 is selected from serine or its analogues.

[0176] In a particular embodiment, P3 is or ,in," "Represents a chiral center, which is selected from the (S) or (R) configuration or a mixture thereof.

[0177] In one particular embodiment, Rc1 is H; in another particular embodiment, Rc1 is OH; in another particular embodiment, Rc1 is CN; in another particular embodiment, Rc1 is NH2; in another particular embodiment, Rc1 is a halogen; in another particular embodiment, Rc1 is O-PG; in another particular embodiment, Rc1 is C. 1-6 Alkyl; in another particular embodiment, Rc1 is C 1-4 Alkyl; in another particular embodiment, Rc1 is C 1-6 alkoxy group; in another particular embodiment, Rc1 is C 1-4 alkoxy group; in another particular embodiment, Rc1 is C 1-6 Haloalkyl; in another particular embodiment, Rc1 is C 3-10 Cycloalkyl; in another particular embodiment, Rc1 is a 3- to 10-membered heterocyclic group; in another particular embodiment, Rc1 is a 5- to 10-membered heteroaryl group, for example, a 5- to 6-membered heteroaryl group; in another particular embodiment, Rc1 is C 6-10 Aryl groups, such as phenyl groups.

[0178] In one particular embodiment, Rc2 is H; in another particular embodiment, Rc2 is OH; in another particular embodiment, Rc2 is CN; in another particular embodiment, Rc2 is NH2; in another particular embodiment, Rc2 is a halogen; in another particular embodiment, Rc2 is O-PG; in another particular embodiment, Rc2 is C. 1-6 Alkyl; in another particular embodiment, Rc2 is C 1-4 Alkyl; in another particular embodiment, Rc2 is C 1-6 alkoxy group; in another particular embodiment, Rc2 is C 1-4 alkoxy group; in another particular embodiment, Rc2 is C 1-6 Halogenated alkyl; in another particular embodiment, Rc2 is C 3-10 Cycloalkyl; in another particular embodiment, Rc2 is a 3- to 10-membered heterocyclic group; in another particular embodiment, Rc2 is a 5- to 10-membered heteroaryl group, for example, a 5- to 6-membered heteroaryl group; in another particular embodiment, Rc2 is C 6-10 Aryl groups, such as phenyl groups.

[0179] In one particular embodiment, Rc3 is H; in another particular embodiment, Rc3 is OH; in another particular embodiment, Rc3 is CN; in another particular embodiment, Rc3 is NH2; in another particular embodiment, Rc3 is a halogen; in another particular embodiment, Rc3 is O-PG; in another particular embodiment, Rc3 is C. 1-6Alkyl; in another particular embodiment, Rc3 is C 1-4 Alkyl; in another particular embodiment, Rc3 is C 1-6 alkoxy group; in another particular embodiment, Rc3 is C 1-4 alkoxy group; in another particular embodiment, Rc3 is C 1-6 Haloalkyl; in another particular embodiment, Rc3 is C 3-10 Cycloalkyl; in another particular embodiment, Rc3 is a 3- to 10-membered heterocyclic group; in another particular embodiment, Rc3 is a 5- to 10-membered heteroaryl group, for example, a 5- to 6-membered heteroaryl group; in another particular embodiment, Rc3 is C 6-10 Aryl groups, such as phenyl groups.

[0180] In a particular embodiment, Rc2 and Rc3 together with the carbon atoms to which they are attached form C 3-10 Cycloalkylene; in another particular embodiment, Rc2 and Rc3 together with the carbon atoms to which they are attached form C 3-7 Cycloalkylene; in another particular embodiment, Rc2 and Rc3 together with the carbon atoms to which they are attached form C 3-5 Cycloalkylene groups; in another particular embodiment, Rc2 and Rc3 together with the carbon atoms to which they are attached form 3- to 10-membered heterocyclic groups.

[0181] In one particular embodiment, Rc1 is not substituted; in another particular embodiment, Rc1 is substituted by one Rc group; in another particular embodiment, Rc1 is substituted by two Rc groups; in yet another particular embodiment, Rc1 is substituted by three Rc groups.

[0182] In one particular embodiment, Rc2 is not substituted; in another particular embodiment, Rc2 is substituted by one Rc group; in another particular embodiment, Rc2 is substituted by two Rc groups; in yet another particular embodiment, Rc2 is substituted by three Rc groups.

[0183] In one particular embodiment, Rc3 is not substituted; in another particular embodiment, Rc3 is substituted by one Rc group; in another particular embodiment, Rc3 is substituted by two Rc groups; in yet another particular embodiment, Rc3 is substituted by three Rc groups.

[0184] In a particular embodiment, c is 0, 1, 2, 3, 4, 5, or 6.

[0185] In one particular embodiment, P3 is not substituted; in another particular embodiment, P3 is substituted by one Rc group; in another particular embodiment, P3 is substituted by two Rc groups; in another particular embodiment, P3 is substituted by three Rc groups; in another particular embodiment, P3 is substituted by four Rc groups; in another particular embodiment, P3 is substituted by five Rc groups.

[0186] In one particular embodiment, Rc is H; in another particular embodiment, Rc is OH; in another particular embodiment, Rc is CN; in another particular embodiment, Rc is NH2; in another particular embodiment, Rc is a halogen; in another particular embodiment, Rc is PG; in another particular embodiment, Rc is C. 1-6 Alkyl; in another particular embodiment, Rc is C 1-4 Alkyl; in another particular embodiment, Rc is C 1-6 Haloalkyl; in another particular embodiment, Rc is C 1-4 Haloalkyl; in another particular embodiment, Rc is C 1-6 Alkoxy groups, such as C 1-4 alkoxy group; in another particular embodiment, Rc is C 1-6 Halogenated alkoxy groups.

[0187] In a particular embodiment, P3 is In another particular embodiment, P3 is In another particular embodiment, P3 is In another particular embodiment, P3 is In another particular embodiment, P3 is In another particular embodiment, P3 is In another particular embodiment, P3 is In another particular embodiment, P3 is In another particular embodiment, P3 is In another particular embodiment, P3 is In another particular embodiment, P3 is .

[0188] P2

[0189] In a particular embodiment, P2 is selected from glycine, serine, or analogues thereof.

[0190] In a particular embodiment, P2 is selected from glycine or its analogues.

[0191] In a particular embodiment, P2 is or ,in," "Represents a chiral center, which is selected from the (S) or (R) configuration or a mixture thereof.

[0192] In one particular embodiment, Rb1 is H; in another particular embodiment, Rb1 is C. 1-6 Alkyl; in another particular embodiment, Rb1 is C 1-4 Alkyl; in another particular embodiment, Rb1 is C 1-6 Halogenated alkyl group; in another particular embodiment, Rb1 is -(CH2). 1-6 -OH; in another particular embodiment, Rb1 is -(CH2). 1-4 -OH.

[0193] In one particular embodiment, Rb2 is H; in another particular embodiment, Rb2 is C. 1-6 Alkyl; in another particular embodiment, Rb2 is C 1-4 Alkyl; in another particular embodiment, Rb2 is C 1-6 Halogenated alkyl group; in another particular embodiment, Rb2 is -(CH2). 1-6 -OH; in another particular embodiment, Rb2 is -(CH2). 1-4 -OH.

[0194] In a particular embodiment, Rb1 and Rb2 together with the carbon atoms to which they are attached form C 3-10 Cycloalkylene; in another particular embodiment, Rb1 and Rb2 together with the carbon atoms to which they are attached form C 3-7 Cycloalkylene; in another particular embodiment, Rb1 and Rb2 together with the carbon atoms to which they are attached form C 3-5 Cycloalkylene groups; in another particular embodiment, Rb1 and Rb2 together with the carbon atoms to which they are attached form 3- to 10-membered heterocyclic groups.

[0195] In one particular embodiment, P2 is not substituted; in another particular embodiment, P2 is substituted by one Rb group; in another particular embodiment, P2 is substituted by two Rb groups; in another particular embodiment, P2 is substituted by three Rb groups; in another particular embodiment, P2 is substituted by four Rb groups; in another particular embodiment, P2 is substituted by five Rb groups.

[0196] In one particular embodiment, Rb is H; in another particular embodiment, Rb is a halogen; in yet another particular embodiment, Rb is PG; in yet another particular embodiment, Rb is C. 1-6 Alkyl; in another particular embodiment, Rb is C 1-6 Halogenated alkyl groups.

[0197] In a particular embodiment, P2 is In another particular embodiment, P2 is In another particular embodiment, P2 is In another particular embodiment, P2 is .

[0198] P1

[0199] In a particular embodiment, P1 is selected from citrulline, arginine, glutamic acid, or analogues thereof.

[0200] In a particular embodiment, P1 is selected from... or ,in," "Represents a chiral center, which is selected from the (S) or (R) configuration or a mixture thereof.

[0201] In a particular embodiment, La is -C 1-8 alkylene-, where -C 1-8 The 1, 2, 3, or 4 non-adjacent carbon atoms in the alkylene group can optionally be replaced by O or S; in another particular embodiment, La is -(CH2). 1-6 -, where -(CH2) 1-6 - One, two, or three non-adjacent carbon atoms in La may optionally be replaced by O; in another particular embodiment, La is -CH2-; in another particular embodiment, La is -CH2CH2-; in another particular embodiment, La is CH2CH2CH2-; in another particular embodiment, La is -CH2OCH2CH2-.

[0202] In one particular embodiment, Xa is a key; in another particular embodiment, Xa is -NR-.

[0203] In one particular embodiment, R is H; in another particular embodiment, R is C. 1-6 Alkyl; in another particular embodiment, R is C 1-4 alkyl.

[0204] In one particular embodiment, R1 is H; in another particular embodiment, R1 is Boc; in yet another particular embodiment, R1 is NR'R'', such as NH2; in yet another particular embodiment, R1 is C. 1-6 Alkyl; in another particular embodiment, R1 is C 1-4 Alkyl; in another particular embodiment, R1 is C 1-6Halogenated alkyl group; in another particular embodiment, R1 is -OR'; in another particular embodiment, R1 is -C(O)R'; in another particular embodiment, R1 is -C(O)OR'; in another particular embodiment, R1 is -C(O)OH; in another particular embodiment, R1 is -C(O)OC 1-4 Alkyl; in another particular embodiment, R1 is -C(NH)NR'R''; in another particular embodiment, R1 is -C(NH)NH2; in another particular embodiment, R1 is -C(O)NR'R''; in another particular embodiment, R1 is -C(O)NH2; in another particular embodiment, R1 is -C(O)NH-C 1-4 Alkyl; in another particular embodiment, R1 is -C(O)NH-C 1-4 Alkylene-OH.

[0205] In one particular embodiment, R' is H; in another particular embodiment, R' is a halogen; in yet another particular embodiment, R' is C. 1-6 Alkyl; in another particular embodiment, R' is C 1-4 Alkyl; in another particular embodiment, R' is C 1-6 Halogenated alkyl; in another particular embodiment, R' is -C 1-6 Alkylene-CN; in another particular embodiment, R' is -C 1-6 Alkylene-NH2; in another particular embodiment, R' is -C 1-6 alkylene-OH; in another particular embodiment, R' is -C 1-4 Alkylene-OH.

[0206] In one particular embodiment, R'' is H; in another particular embodiment, R'' is a halogen; in yet another particular embodiment, R'' is C. 1-6 Alkyl; in another particular embodiment, R'' is C 1-4 Alkyl; in another particular embodiment, R'' is C 1-6 Halogenated alkyl; in another particular embodiment, R'' is -C 1-6 Alkylene-CN; in another particular embodiment, R'' is -C 1-6 Alkylene-NH2; in another particular embodiment, R'' is -C 1-6 alkylene-OH; in another particular embodiment, R'' is -C 1-4 Alkylene-OH.

[0207] In one particular embodiment, P1 is not substituted; in another particular embodiment, P1 is substituted by one Ra group; in another particular embodiment, P1 is substituted by two Ra groups; in another particular embodiment, P1 is substituted by three Ra groups; in another particular embodiment, P1 is substituted by four Ra ​​groups; in another particular embodiment, P1 is substituted by five Ra groups.

[0208] In one particular embodiment, Ra is H; in another particular embodiment, Ra is a halogen; in yet another particular embodiment, Ra is PG; in yet another particular embodiment, Ra is C. 1-6 Alkyl; in another particular embodiment, Ra is C 1-6 Halogenated alkyl groups.

[0209] In a particular embodiment, P1 is In another particular embodiment, P1 is In another particular embodiment, P1 is In another particular embodiment, P1 is In another particular embodiment, P1 is In another particular embodiment, P1 is In another particular embodiment, P1 is In another particular embodiment, P1 is In another particular embodiment, P1 is In another particular embodiment, P1 is In another particular embodiment, P1 is In another particular embodiment, P1 is In another particular embodiment, P1 is In another particular embodiment, P1 is .

[0210] In some embodiments, PG is selected from: trimethylsilyl group (TMS), triethylsilyl group (TES), dimethylisopropylsilyl group (DMIPS), diethylisopropylsilyl group (DEIPS), tert-butyldimethylsilyl group (TBDMS), tert-butyldiphenylsilyl group (TBDPS), triisopropylsilyl group (TIPS), acetyl group (Ac), chloroacetyl group, dichloroacetyl group, trichloroacetyl group, trifluoroacetyl group (TFA), benzoyl group, p-methoxybenzoyl group, 9-fluorenylmethoxycarbonyl group (Fmoc), allyloxycarbonyl group (All). oc), 2,2,2-trichloroethoxycarbonyl (Troc), carboxybenzyl (Cbz), tert-butoxycarbonyl (Boc), benzyl (Bn), p-methoxybenzyl (PMB), allyl, triphenylmethyl (Tr), bis-p-methoxytriphenylmethyl (DMTr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), p-methoxybenzyloxymethyl (PMBM), -C(O)CH2CH2C(O)OH or 4,4'-dimethoxytriphenylmethyl.

[0211] Y

[0212] In a particular embodiment, Y is a spacer subunit.

[0213] In a particular embodiment, Y includes a self-destructing spacer subunit.

[0214] In a particular embodiment, Y is In another particular embodiment, Y is .

[0215] In a particular embodiment, R Y1 It is H; in another particular embodiment, R Y1 It is a halogen; in another particular embodiment, R Y1 It is OH; in another particular embodiment, R Y1 It is CN; in another particular embodiment, R Y1 It is C 1-6 Alkyl, such as C 1-4 Alkyl; in another particular embodiment, R Y1 It is C 1-6 Alkoxy groups, such as C 1-4 Alkoxy; in another particular embodiment, R Y1 It is C 1-6 Haloalkyl, such as C 1-4 Halogenated alkyl; in another particular embodiment, R Y1 It is C 1-6 Halogenated alkoxy groups, such as C 1-4Halogenated alkoxy groups.

[0216] In a particular embodiment, R Y2 It is H; in another particular embodiment, R Y2 It is a halogen; in another particular embodiment, R Y2 It is OH; in another particular embodiment, R Y2 It is CN; in another particular embodiment, R Y2 It is C 1-6 Alkyl, such as C 1-4 Alkyl; in another particular embodiment, R Y2 It is C 1-6 Alkoxy groups, such as C 1-4 Alkoxy; in another particular embodiment, R Y2 It is C 1-6 Haloalkyl, such as C 1-4 Halogenated alkyl; in another particular embodiment, R Y2 It is C 1-6 Halogenated alkoxy groups, such as C 1-4 Halogenated alkoxy groups.

[0217] In a particular embodiment, p is 0, 1, or 2.

[0218] In a particular embodiment, q is 0, 1, 2, 3, or 4.

[0219] In a particular embodiment, Y is (AM); in another particular embodiment, Y is (PABC).

[0220] D

[0221] In a particular embodiment, the pharmaceutical portion D is selected from the group consisting of: microtubule inhibitors and microtubule polymerization inhibitors, antibiotics, DNA synthesis inhibitors, topoisomerase I inhibitors, immunomodulators, and chelating ligands.

[0222] In a particular embodiment, the pharmaceutical part D is a tubulin inhibitor and a microtubule polymerization inhibitor selected from the group consisting of: olistatins (e.g., MMAE, MMAF, and MMAD), maytansines (e.g., DM1, DM2, DM3, and DM4), tubulolysin, cryptophycins, eribulin, and rhizoxin.

[0223] In a particular embodiment, the pharmaceutical part D is an antibiotic selected from the group consisting of calicheamicin, doxorubicin, and anthracyclines.

[0224] In a particular embodiment, the pharmaceutical part D is a DNA synthesis inhibitor selected from the group consisting of duocarmycin, PBD (pyrrolobenzodiazepine), and IGN (indolinobenzodiazepine).

[0225] In a particular embodiment, the pharmaceutical part D is a topoisomerase I inhibitor selected from the group consisting of camptothecin and camptothecin analogues (e.g., DXd, SN38, and eczetidine).

[0226] In a particular embodiment, the pharmaceutical part D is an immunomodulator selected from the group consisting of: TLR7 agonists, TLR8 agonists, STING agonists, and RIG-I agonists.

[0227] In a particular embodiment, the pharmaceutical portion D is In another particular embodiment, the pharmaceutical portion D is... In another particular embodiment, the pharmaceutical portion D is... .

[0228] In a particular embodiment, pharmaceutical portion D is derived from a compound having the formula (DI) or (D-II):

[0229] (DI) or (D-II)

[0230] The variables Rd1, Rd2, and Rd3 are defined as in the context.

[0231] In a particular embodiment, pharmaceutical portion D is derived from the following compounds:

[0232] , , , , , , , , , , , or .

[0233] In a particular embodiment, the pharmaceutical portion D is selected from compounds having the formula (DI) or (D-II):

[0234] (D-III) (D-IV)

[0235] in, Indicates the attachment site with the rest of the compound;

[0236] Variables Rd1, Rd2, and Rd3' are defined as in the context.

[0237] In a particular embodiment, pharmaceutical portion D is selected from the following compounds:

[0238] , , , , , , , , , , , or .

[0239] In a particular embodiment, the pharmaceutical portion D is .

[0240] Rd1 and Rd2

[0241] In a particular embodiment, Rd1 is C 1-6 Alkyl; in another particular embodiment, Rd1 is C 1-6 alkoxy group; in another particular embodiment, Rd1 is C 1-4 Alkyl; in another particular embodiment, Rd1 is C 1-4 Alkyl groups, such as Me or OMe.

[0242] In one particular embodiment, Rd2 is a halogen; in another particular embodiment, Rd2 is F.

[0243] In a particular embodiment, Rd1 and Rd2 together with the carbon atoms to which they are attached form a 5-10 membered heterocyclic group or a 5-10 membered heteroarylene group, which is optionally surrounded by one, two, or three atoms selected from H, D, halogens, and C. 1-6 Alkyl substituents; in another particular embodiment, Rd1 and Rd2 together with the carbon atoms to which they are attached form a 5-6 membered heterocyclic cycloidene or a 5-6 membered heteroarylene (preferably forming a 5-6 membered heterocyclic cycloidene), which is optionally replaced by one, two or three atoms selected from H, D, halogens and C. 1-4 Alkyl substituents; in another particular embodiment, Rd1, Rd2 together with the carbon atoms to which they are attached form , or .

[0244] Rd3

[0245] In a particular embodiment, Rd3 is -(CH2). p -ORd a In another particular embodiment, Rd3 is -(CH2). p -C(O)ORd a In another particular embodiment, Rd3 is -(CH2). p -NHC(O)-Rd a In another particular embodiment, Rd3 is -(CH2). p -CH2NRd b Rd c .

[0246] In a particular embodiment, Rd3 is In another particular embodiment, Rd3 is In another particular embodiment, Rd3 is In another particular embodiment, Rd3 is In another particular embodiment, Rd3 is In another particular embodiment, Rd3 is ;

[0247] In a particular embodiment, Rd3 is In another particular embodiment, Rd3 is In another particular embodiment, Rd3 is In another particular embodiment, Rd3 is In another particular embodiment, Rd3 is In another particular embodiment, Rd3 is In another particular embodiment, Rd3 is In another particular embodiment, Rd3 is In another particular embodiment, Rd3 is In another particular embodiment, Rd3 is In another particular embodiment, Rd3 is .

[0248] Rd3'

[0249] In a particular embodiment, Rd3' is -(CH2). p -ORd a In another particular embodiment, Rd3' is -(CH2). p -C(O)ORd a In another particular embodiment, Rd3' is -(CH2). p -NHC(O)-Rd aIn another particular embodiment, Rd3' is -(CH2). p -CH2NRd b Rd c '-.

[0250] In a particular embodiment, Rd3' is In another particular embodiment, Rd3' is In another particular embodiment, Rd3' is In another particular embodiment, Rd3' is In another particular embodiment, Rd3' is In another particular embodiment, Rd3' is .

[0251] In a particular embodiment, Rd3' is In another particular embodiment, Rd3' is In another particular embodiment, Rd3' is In another particular embodiment, Rd3' is In another particular embodiment, Rd3' is In another particular embodiment, Rd3' is In another particular embodiment, Rd3' is In another particular embodiment, Rd3' is In another particular embodiment, Rd3' is In another particular embodiment, Rd3' is In another particular embodiment, Rd3' is .

[0252] Rd a 、Rd b and Rd c

[0253] In a particular embodiment, Rd a It is H; in another particular embodiment, Rd a It is C 1-6 Alkyl; in another particular embodiment, Rd a It is C 1-6 alkylene-OH; in another particular embodiment, Rd a It is C 1-6 Alkylene-NH2; in another particular embodiment, Rd a It is CH3; in another particular embodiment, Rd a It is CH2OH; in another particular embodiment, Rd aIt is CH2CH2OH; in another particular embodiment, Rd a It is CH2CH2NH2.

[0254] In a particular embodiment, Rd b It is H; in another particular embodiment, Rd b It is C 1-6 Alkyl groups, such as CH3.

[0255] In a particular embodiment, Rd c It is H; in another particular embodiment, Rd c It is C 1-6 Alkyl groups, such as CH3.

[0256] Rd a 'and Rd c '

[0257] In a particular embodiment, Rd a 'is a key; in another particular embodiment, Rd a 'Is C 1-6 Alkylene; in another particular embodiment, Rd a 'is-C 1-6 alkylene-O-; in another particular embodiment, Rd a 'is-C 1-6 alkylene-NH-; in another particular embodiment, Rd a 'is -CH2-; in another particular embodiment, Rd a 'is -CH2O-; in another particular embodiment, Rd a 'is -CH2CH2O-; in another particular embodiment, Rd a 'It is -CH2CH2NH-.

[0258] In a particular embodiment, Rd c 'is a key; in another particular embodiment, C 1-6 Alkylene, for example -CH2-.

[0259] p

[0260] In a particular embodiment, each p is independently 0, 1, 2, 3, 4, 5, or 6.

[0261] Any technical solution or any combination thereof in any of the above special embodiments can be combined with any technical solution or any combination thereof in other special embodiments. For example, any technical solution or any combination thereof in L can be combined with any technical solution or any combination thereof in Y, P, and D. This disclosure is intended to include all combinations of such technical solutions, and for the sake of brevity, they will not be listed in detail here.

[0262] 1.2 Examples

[0263] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein:

[0264] L is represented by L1-L2-X-;

[0265] L1 is the ligand covalently bound portion and can be alternatively selected from... and ;

[0266] -L2-X- is an extended subunit;

[0267] L2 is selected from C 1-10 Alkylene, -(C 1-4 alkylene-O) 1-6 -、-(OC 1-4 Alkylene) 1-6 -、-C 2-10 imide and C 2-10 Ethyne group;

[0268] X either does not exist or is -C(O)-;

[0269] L can be arbitrarily divided by 1, 2, 3, 4 or 5 Rs. L Group substitution;

[0270] R L Selected from H, D, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups.

[0271] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein:

[0272] L is represented by L1-L2-X-;

[0273] L1 is selected from and ;

[0274] L2 is selected from C 1-10 Alkylene, -(CH2) 0-4 -(CH2CH2-O) 1-6 -(CH2) 0-4 -、-(CH2) 0-4 -(O-CH2CH2) 1-6 -(CH2) 0-4 -、C 2-10 imide and C 2-10 Ethyne group;

[0275] X either does not exist or is -C(O)-;

[0276] L can be optionally divided by 1, 2 or 3 Rs L Group substitution;

[0277] R L Selected from H, D, C 1-4 Alkyl and C 1-4 Halogenated alkyl groups.

[0278] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein:

[0279] L is selected from , and ;

[0280] m and n are independently selected from 1, 2, 3, 4, 5, 6, 7 or 8;

[0281] Alternatively, m can be selected from 4, 5, or 6, and can be 5;

[0282] Alternatively, n can be selected from 2, 3, or 4, and 3 is an alternative.

[0283] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein:

[0284] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0285] Among them, P3 is ,

[0286] Rc1, Rc2, and Rc3 are independently selected from H, OH, CN, NH2, halogens, O-PG, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 3-10 Cycloalkyl, 3- to 10-membered heterocyclic, 5- to 10-membered heteroaryl and C 6-10 Aryl;

[0287] Or Rc2 and Rc3 together with the carbon atoms to which they are attached form C 3-10 Cycloalkylene or 3- to 10-membered heterocyclic cycloidene groups;

[0288] c is 0, 1, 2, 3 or 4;

[0289] P3 may optionally be further substituted with 1, 2, 3, 4 or 5 Rc groups;

[0290] Rc is selected from H, OH, CN, NH2, halogens, PG, and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0291] P2 is Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl, C 1-6 Halogenated alkyl groups and -(CH2) 1-6 -OH, or Rb1 and Rb2 together with the carbon atoms to which they are attached, form C 3-10 Cycloalkylene or 3- to 10-membered heterocyclic cycloidene groups;

[0292] P2 may optionally be further substituted with 1, 2, 3, 4 or 5 Rb groups;

[0293] Rb is selected from H, halogens, PG, and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0294] P1 is selected from ;

[0295] La is -C 1-8 alkylene-, where -C 1-8 The 1, 2, 3 or 4 non-adjacent carbon atoms in the alkylene group can be optionally replaced by O or S;

[0296] Xa is a bond or -NR-;

[0297] R is H or -C 1-4 alkyl;

[0298] R1 is selected from H, Boc, and C. 1-6 Alkyl, -OR', NR'R'', -C(O)R', -C(O)OR', -C(NH)NR'R'' and -C(O)NR'R'';

[0299] R' and R'' are independently selected from H, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, -C 1-6 Alkylene-CN, -C 1-6 Alkyl-NH2 and -C 1-6 alkylene-OH;

[0300] P1 may optionally be further substituted with 1, 2, 3, 4 or 5 Ra groups;

[0301] Ra is selected from H, halogen, PG, C 1-6 Alkyl and C1-6 Halogenated alkyl groups;

[0302] “ "Represents a chiral center, which is selected from the (S) or (R) configuration;

[0303] PG is selected from: trimethylsilyl group (TMS), triethylsilyl group (TES), dimethylisopropylsilyl group (DMIPS), diethylisopropylsilyl group (DEIPS), tert-butyldimethylsilyl group (TBDMS), tert-butyldiphenylsilyl group (TBDPS), triisopropylsilyl group (TIPS), acetyl group (Ac), chloroacetyl group, dichloroacetyl group, trichloroacetyl group, trifluoroacetyl group (TFA), benzoyl group, p-methoxybenzoyl group, 9-fluorenylmethoxycarbonyl group (Fmoc), allyloxycarbonyl group (Alloc). 2,2,2-Trichloroethoxycarbonyl (Troc), carboxybenzyl (Cbz), tert-butoxycarbonyl (Boc), benzyl (Bn), p-methoxybenzyl (PMB), allyl, triphenylmethyl (Tr), bis-p-methoxytriphenylmethyl (DMTr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), p-methoxybenzyloxymethyl (PMBM), -C(O)CH2CH2C(O)OH or 4,4'-dimethoxytriphenylmethyl.

[0304] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein:

[0305] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0306] Among them, P3 is ,

[0307] Rc1 is selected from OH, O-PG, and C. 1-4 The alkoxy groups, Rc2 and Rc3, are independently selected from H, halogens, and C. 1-4 Alkyl and C 6-10 Aryl groups (such as phenyl), or Rc2 and Rc3, together with the carbon atoms to which they are attached, form C 3-5 Cycloalkylene;

[0308] Rc1, Rc2 and Rc3 can be independently and optionally replaced by 1, 2 or 3 Rc;

[0309] Rc is selected from H, OH, C 1-4 Alkyl and C 1-4Halogenated alkyl groups;

[0310] c is 0, 1, or 2;

[0311] P2 is Rb1 and Rb2 are independently selected from H and C. 1-4 Alkyl groups and -(CH2) 1-4 -OH, or Rb1 and Rb2 together with the carbon atoms to which they are attached, form C 3-5 Cycloalkylene;

[0312] P1 is selected from ;

[0313] La is -(CH2) 1-6 -, where -(CH2) 1-6 The 1, 2, or 3 non-adjacent carbon atoms in - can be optionally replaced by O; alternatively, La is selected from -CH2-, -CH2CH2-, CH2CH2CH2-, and -CH2OCH2CH2-;

[0314] Xa is a bond or -NR-;

[0315] R is H or -C 1-4 alkyl;

[0316] R1 is selected from H, Boc, NR'R'', and C. 1-4 Alkyl groups, -C(NH)NR'R'', -C(O)NR'R'', and -C(O)OR';

[0317] R' and R'' are independently selected from H and -C. 1-4 Alkyl and -C 1-4 Alkylene-OH.

[0318] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein:

[0319] R1 is selected from Boc, NH2, and C. 1-4 Alkyl groups, -C(NH)NH2, -C(O)NH2, -C(O)NH-C 1-4 Alkyl group, -C(O)NH-C 1-4 Alkylenes -OH, -C(O)OH and -C(O)OC 1-4 alkyl;

[0320] “ "Represents a chiral center, which is selected from the (S) or (R) configuration;

[0321] Alternatively,

[0322] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0323] P3 is selected from: , , , , , and ;

[0324] P2 is selected from: , , and ;

[0325] P1 is selected from: , , , , , , , , , , , , and .

[0326] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein:

[0327] Y is selected from: and ;

[0328] R Y1 Selected from H, halogens, OH, CN, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl and C 1-6 Halogenated alkoxy groups;

[0329] R Y2 Selected from H, halogens, OH, CN, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl and C 1-6 Halogenated alkoxy groups;

[0330] p is 0, 1, or 2;

[0331] q is 0, 1, 2, 3, or 4;

[0332] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein:

[0333] Y is selected from: and ;

[0334] R Y1 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0335] R Y2 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0336] p is 0, 1, or 2;

[0337] q is 0, 1, or 2.

[0338] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein:

[0339] Y is selected from: (AM) and (PABC).

[0340] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein:

[0341] The drug part D is selected from the following groups: microtubule inhibitors and microtubule polymerization inhibitors, antibiotics, DNA synthesis inhibitors and topoisomerase I inhibitors.

[0342] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein pharmaceutical part D is a tubulin inhibitor and a microtubule polymerization inhibitor selected from the group consisting of: olistatins (e.g., MMAE, MMAF, and MMAD), maytansines (e.g., DM1, DM2, DM3, and DM4), tubulolysin, candidacin, and rhizobium.

[0343] This antibiotic was selected from the group consisting of: chalcone, doxorubicin, and anthracyclines;

[0344] The DNA synthesis inhibitor was selected from the group consisting of: betamethasone, PBD (pyrrolobenzodiazepine), and IGN (indolinobenzodiazepine).

[0345] The topoisomerase I inhibitors are selected from the group consisting of: camptothecin analogs (e.g., DXd, SN38 and ixotecan).

[0346] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein:

[0347] D is selected from: , and .

[0348] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein:

[0349] L is represented by L1-L2-X-;

[0350] L1 is the ligand covalently bound portion and can be alternatively selected from... and ;

[0351] -L2-X- is an extended subunit;

[0352] L2 is selected from C 1-10 Alkylene, -(C 1-4 alkylene-O) 1-6 -、-(OC 1-4 Alkylene) 1-6 -、-C 2-10 imide and C 2-10 Ethyne group;

[0353] X either does not exist or is -C(O)-;

[0354] L can be arbitrarily divided by 1, 2, 3, 4 or 5 Rs. L Group substitution;

[0355] R L Selected from H, D, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0356] Y is selected from: and ;

[0357] R Y1 Selected from H, halogens, OH, CN, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl and C 1-6 Halogenated alkoxy groups;

[0358] RY2 Selected from H, halogens, OH, CN, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl and C 1-6 Halogenated alkoxy groups;

[0359] p is 0, 1, or 2;

[0360] q is 0, 1, 2, 3, or 4;

[0361] Part D of this drug is selected from the following groups: microtubule inhibitors and microtubule polymerization inhibitors, antibiotics, DNA synthesis inhibitors and topoisomerase I inhibitors;

[0362] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0363] Among them, P3 is ,

[0364] Rc1, Rc2, and Rc3 are independently selected from H, OH, CN, NH2, halogens, O-PG, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 3-10 Cycloalkyl, 3- to 10-membered heterocyclic, 5- to 10-membered heteroaryl and C 6-10 Aryl;

[0365] Or Rc2 and Rc3 together with the carbon atoms to which they are attached form C 3-10 Cycloalkylene or 3- to 10-membered heterocyclic cycloidene groups;

[0366] c is 0, 1, 2, 3 or 4;

[0367] P3 may optionally be further substituted with 1, 2, 3, 4 or 5 Rc groups;

[0368] Rc is selected from H, OH, CN, NH2, halogens, PG, and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0369] P2 is Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl, C 1-6 Halogenated alkyl groups and -(CH2) 1-6 -OH, or Rb1 and Rb2 together with the carbon atoms to which they are attached, form C 3-10 Cycloalkylene or 3- to 10-membered heterocyclic cycloidene groups;

[0370] P2 may optionally be further substituted with 1, 2, 3, 4 or 5 Rb groups;

[0371] Rb is selected from H, halogens, PG, and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0372] P1 is selected from ;

[0373] La is -C 1-8 alkylene-, where -C 1-8 The 1, 2, 3 or 4 non-adjacent carbon atoms in the alkylene group can be optionally replaced by O or S;

[0374] Xa is a bond or -NR-;

[0375] R is H or -C 1-4 alkyl;

[0376] R1 is selected from H, Boc, and C. 1-6 Alkyl, -OR', NR'R'', -C(O)R', -C(O)OR', -C(NH)NR'R'' and -C(O)NR'R'';

[0377] R' and R'' are independently selected from H, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, -C 1-6 Alkylene-CN, -C 1-6 Alkyl-NH2 and -C 1-6 alkylene-OH;

[0378] P1 may optionally be further substituted with 1, 2, 3, 4 or 5 Ra groups;

[0379] Ra is selected from H, halogen, PG, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0380] “ "Represents a chiral center, which is selected from the (S) or (R) configuration;

[0381] PG is selected from: trimethylsilyl group (TMS), triethylsilyl group (TES), dimethylisopropylsilyl group (DMIPS), diethylisopropylsilyl group (DEIPS), tert-butyldimethylsilyl group (TBDMS), tert-butyldiphenylsilyl group (TBDPS), triisopropylsilyl group (TIPS), acetyl group (Ac), chloroacetyl group, dichloroacetyl group, trichloroacetyl group, trifluoroacetyl group (TFA), benzoyl group, p-methoxybenzoyl group, 9-fluorenylmethoxycarbonyl group (Fmoc), allyloxycarbonyl group (Alloc). 2,2,2-Trichloroethoxycarbonyl (Troc), carboxybenzyl (Cbz), tert-butoxycarbonyl (Boc), benzyl (Bn), p-methoxybenzyl (PMB), allyl, triphenylmethyl (Tr), bis-p-methoxytriphenylmethyl (DMTr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), p-methoxybenzyloxymethyl (PMBM), -C(O)CH2CH2C(O)OH or 4,4'-dimethoxytriphenylmethyl.

[0382] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein:

[0383] L is represented by L1-L2-X-;

[0384] L1 is selected from and ;

[0385] L2 is selected from C 1-10 Alkylene, -(CH2) 0-4 -(CH2CH2-O) 1-6 -(CH2) 0-4 -、-(CH2) 0-4 -(O-CH2CH2) 1-6 -(CH2) 0-4 -、C 2-10 imide and C 2-10 Ethyne group;

[0386] X either does not exist or is -C(O)-;

[0387] L can be optionally divided by 1, 2 or 3 Rs L Group substitution;

[0388] R L Selected from H, D, C 1-4 Alkyl and C 1-4 Halogenated alkyl groups;

[0389] Y is selected from: and ;

[0390] R Y1 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0391] R Y2 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0392] Part D of this drug is selected from the following groups: microtubule inhibitors and microtubule polymerization inhibitors, antibiotics, DNA synthesis inhibitors and topoisomerase I inhibitors;

[0393] The microtubule inhibitors and microtubule polymerization inhibitors are selected from the following groups: olistatins (e.g., MMAE, MMAF, and MMAD), maytansine (e.g., DM1, DM2, DM3, and DM4), tubulosyntheticin, candidin, and rhizobium.

[0394] This antibiotic was selected from the group consisting of: chalcone, doxorubicin, and anthracyclines;

[0395] The DNA synthesis inhibitor was selected from the group consisting of: betamethasone, PBD (pyrrolobenzodiazepine), and IGN (indolinobenzodiazepine).

[0396] Topoisomerase I inhibitors are selected from the group consisting of: camptothecin analogs (e.g., DXd, SN38, and eczetidine);

[0397] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0398] Among them, P3 is ,

[0399] Rc1 is selected from OH, O-PG, and C. 1-4 The alkoxy groups, Rc2 and Rc3, are independently selected from H, halogens, and C. 1-4 Alkyl and C 6-10 Aryl groups (such as phenyl), or Rc2 and Rc3, together with the carbon atoms to which they are attached, form C 3-5 Cycloalkylene;

[0400] Rc1, Rc2 and Rc3 can be independently and optionally replaced by 1, 2 or 3 Rc;

[0401] Rc is selected from H, OH, C 1-4 Alkyl and C 1-4 Halogenated alkyl groups;

[0402] c is 0, 1, or 2;

[0403] P2 is Rb1 and Rb2 are independently selected from H and C. 1-4 Alkyl groups and -(CH2) 1-4 -OH, or Rb1 and Rb2 together with the carbon atoms to which they are attached, form C 3-5 Cycloalkylene;

[0404] P1 is selected from ;

[0405] La is -(CH2) 1-6 -, where -(CH2) 1-6 The 1, 2, or 3 non-adjacent carbon atoms in - can be optionally replaced by O; alternatively, La is selected from -CH2-, -CH2CH2-, CH2CH2CH2-, and -CH2OCH2CH2-;

[0406] Xa is a bond or -NR-;

[0407] R is H or -C 1-4 alkyl;

[0408] R1 is selected from H, Boc, NR'R'', and C. 1-4 Alkyl groups, -C(NH)NR'R'', -C(O)NR'R'', and -C(O)OR';

[0409] R' and R'' are independently selected from H and -C. 1-4 Alkyl and -C 1-4 alkylene-OH;

[0410] Alternatively, R1 can be selected from Boc, NH2, and C. 1-4 Alkyl groups, -C(NH)NH2, -C(O)NH2, -C(O)NH-C 1-4 Alkyl group, -C(O)NH-C 1-4 Alkylenes -OH, -C(O)OH and -C(O)OC 1-4 alkyl;

[0411] “ "Represents a chiral center, which is selected from the (S) or (R) configuration.

[0412] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein:

[0413] L is selected from , and ;

[0414] m and n are independently selected from 1, 2, 3, 4, 5, 6, 7 or 8;

[0415] Alternatively, m can be selected from 4, 5, or 6, and can be 5;

[0416] Alternatively, n can be selected from 2, 3, or 4, and 3 is an alternative.

[0417] Y is selected from: (AM) and (PABC);

[0418] D is selected from: , and ;

[0419] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0420] P3 is selected from: , , , , , and ;

[0421] P2 is selected from: , , and ;

[0422] P1 is selected from: , , , , , , , , , , , , and .

[0423] In some embodiments, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is selected from:

[0424] ;

[0425] ;

[0426] ;

[0427] ;

[0428] ;

[0429] ;as well as

[0430] ,

[0431] Where m, n, P3, P2, and P1 are as defined in the context;

[0432] Alternatively,

[0433] m is selected from 4, 5 or 6, and can be 4 or 5;

[0434] n is selected from 2, 3, or 4, and can be 3 instead.

[0435] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein the compound is MC-VGR-DXD:

[0436] .

[0437] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-SGR-DXD:

[0438] .

[0439] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-VGR-PABC-MMAE:

[0440] .

[0441] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-SGR-PABC-MMAE:

[0442] .

[0443] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-SGC-DXD:

[0444] .

[0445] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-SAC-DXD:

[0446] .

[0447] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MsP-SGC-DXD:

[0448] .

[0449] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-S(tBu)GC-DXD:

[0450] .

[0451] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-SGOrn(Boc)-DXD:

[0452] .

[0453] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-SGC-PABC-Ecinotecan:

[0454] .

[0455] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein the compound is MC-SGO(DEt)-PABC-ecientam:

[0456] .

[0457] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein the compound is MC-SGE-PABC-Ecinotecan:

[0458] .

[0459] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein the compound is MC-SGE(tBu)-PABC-ecientam:

[0460] .

[0461] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein the compound is MC-SGC(Me)-PABC-eccinotecan:

[0462] .

[0463] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein the compound is MC-SGC(Peg)-PABC-Ecinotecan:

[0464] .

[0465] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-TGC-PABC-Ecinotecan:

[0466] .

[0467] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein the compound is MC-SGhC(O)-PABC-ecientam:

[0468] .

[0469] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-YSR-PABC-Ecinotecan:

[0470] .

[0471] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-SGE-DXD:

[0472] .

[0473] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-SGC(Peg)-DXD:

[0474] .

[0475] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-TSR-DXD:

[0476] ,and

[0477] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-YSR-DXD:

[0478] .

[0479] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-SGO-DXD:

[0480] .

[0481] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-SGK-DXD:

[0482] .

[0483] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is Mal-PEG4-VGR-DXD:

[0484] .

[0485] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is Mal-PEG4-SGR-DXD:

[0486] .

[0487] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein the compound is MC-SGhR-DXD:

[0488] .

[0489] In a particular embodiment, this disclosure relates to a compound having formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MC-SGN-DXD:

[0490] .

[0491] 1.3 Preferred Embodiments

[0492] In some embodiments, this disclosure relates to a compound having formula (X) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof:

[0493] SD (X)

[0494] in,

[0495] S is a linker, and the linker contains a peptide-cleavable unit that contains a tripeptide having the sequence -P3-P2-P1-.

[0496] P3 is selected from serine, tyrosine, or their analogues;

[0497] P2 is selected from glycine, serine, or their analogues;

[0498] P1 is selected from citrulline, arginine, glutamic acid or their analogues;

[0499] D represents the drug component.

[0500] In some embodiments, this disclosure relates to a compound having formula (X) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein,

[0501] The connector S further includes a cuttable connector or a non-cuttable connector;

[0502] Alternatively, the cleavable linker may comprise an acid-instable linker, a hydrophilic linker, a protease-sensitive linker, a light-instable linker, a hydrazone linker, a dimethyl linker, or a disulfide-containing linker.

[0503] In some embodiments, this disclosure relates to compounds having formula (X), wherein the compound has the structure of formula (I), or a pharmaceutically acceptable salt, enantiomer, diastereomer, or mixture thereof:

[0504] LPYD (I)

[0505] L is represented by L1-L2-X-;

[0506] L1 is the ligand covalently bound portion.

[0507] -L2-X- is an extended subunit;

[0508] Y is the spacer subunit;

[0509] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0510] P3 is selected from serine, tyrosine, or their analogues;

[0511] P2 is selected from glycine, serine, or their analogues;

[0512] P1 is selected from citrulline, arginine, glutamic acid, or their analogues.

[0513] D represents the drug component.

[0514] In some embodiments, this disclosure relates to compounds having formula (X) or formula (I), or pharmaceutically acceptable salts, enantiomers, diastereomers, or mixtures thereof:

[0515] Among them, P3 is ,

[0516] Rc1, Rc2, and Rc3 are independently selected from H, OH, CN, NH2, halogens, O-PG, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 3-10 Cycloalkyl, 3- to 10-membered heterocyclic, 5- to 10-membered heteroaryl and C 6-10 Aryl;

[0517] Or Rc2 and Rc3 together with the carbon atoms to which they are attached form C 3-10 Cycloalkylene or 3- to 10-membered heterocyclic cycloidene groups;

[0518] c is 0, 1, 2, 3 or 4;

[0519] P3 may optionally be further substituted with 1, 2, 3, 4 or 5 Rc groups;

[0520] Rc is selected from H, OH, CN, NH2, halogens, PG, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl and C 1-6 The haloalkoxy group can be alternatively selected from H, OH, CN, NH2, halogen, PG, C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0521] P2 is Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl, C 1-6 Halogenated alkyl groups and -(CH2) 1-6 -OH, or Rb1 and Rb2 together with the carbon atoms to which they are attached, form C 3-10 Cycloalkylene or 3- to 10-membered heterocyclic cycloidene groups;

[0522] P2 may optionally be further substituted with 1, 2, 3, 4 or 5 Rb groups;

[0523] Rb is selected from H, halogens, PG, and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0524] P1 is selected from ;

[0525] La is -C 1-8 alkylene-, where -C 1-8 The 1, 2, 3 or 4 non-adjacent carbon atoms in the alkylene group can be optionally replaced by O or S;

[0526] Xa is a bond or -NR-;

[0527] R is H or -C 1-4 alkyl;

[0528] R1 is selected from H, Boc, and C. 1-6 Alkyl groups, -OR', NR'R'', -C(O)R', -C(O)OR', -C(NH)NR'R'', and -C(O)NR'R'', preferably selected from H, Boc, C 1-6 Alkyl, -OR', -C(O)R', -C(O)OR', -C(NH)NR'R'' and -C(O)NR'R'';

[0529] R' and R'' are independently selected from H, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, -C 1-6 Alkylene-CN, -C 1-6 Alkyl-NH2 and -C 1-6 alkylene-OH;

[0530] P1 may optionally be further substituted with 1, 2, 3, 4 or 5 Ra groups;

[0531] Ra is selected from H, halogen, PG, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0532] “ "Represents a chiral center, which is selected from the (S) or (R) configuration;

[0533] PG is selected from: trimethylsilyl group (TMS), triethylsilyl group (TES), dimethylisopropylsilyl group (DMIPS), diethylisopropylsilyl group (DEIPS), tert-butyldimethylsilyl group (TBDMS), tert-butyldiphenylsilyl group (TBDPS), triisopropylsilyl group (TIPS), acetyl group (Ac), chloroacetyl group, dichloroacetyl group, trichloroacetyl group, trifluoroacetyl group (TFA), benzoyl group, p-methoxybenzoyl group, 9-fluorenylmethoxycarbonyl group (Fmoc), allyloxycarbonyl group (Alloc), 2,2 2-Trichloroethoxycarbonyl (Troc), carboxybenzyl (Cbz), tert-butoxycarbonyl (Boc), benzyl (Bn), p-methoxybenzyl (PMB), allyl, triphenylmethyl (Tr), bis-p-methoxytriphenylmethyl (DMTr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), p-methoxybenzyloxymethyl (PMBM), -C(O)CH2CH2C(O)OH or 4,4'-dimethoxytriphenylmethyl, alternatively,

[0534] Alternatively,

[0535] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0536] Among them, P3 is ,

[0537] Rc1 is selected from H, OH, O-PG, and C. 1-4 Alkyl groups, Rc2 and Rc3, are independently selected from H, halogens, C. 1-4Alkyl, 5- to 10-membered heteroaryl (e.g., 5- to 6-membered heteroaryl) and C 6-10 Aryl groups (such as phenyl), or Rc2 and Rc3, together with the carbon atoms to which they are attached, form C 3-5 Cycloalkylene;

[0538] Rc1, Rc2 and Rc3 can be independently and optionally replaced by 1, 2 or 3 Rc;

[0539] Rc is selected from H, OH, C 1-4 Alkyl, C 1-4 Alkoxy and C 1-4 Haloalkyl groups, which may be selected from H, OH, C. 1-4 Alkyl and C 1-4 Halogenated alkyl groups;

[0540] c is 0, 1, or 2;

[0541] P2 is Rb1 and Rb2 are independently selected from H and C. 1-4 Alkyl groups and -(CH2) 1-4 -OH, or Rb1 and Rb2 together with the carbon atoms to which they are attached, form C 3-5 Cycloalkylene;

[0542] P1 is selected from ;

[0543] La is -(CH2) 1-6 -, where -(CH2) 1-6 The 1, 2, or 3 non-adjacent carbon atoms in - can be optionally replaced by O; alternatively, La is selected from -CH2-, -CH2CH2-, CH2CH2CH2-, CH2CH2CH2CH2-, and -CH2OCH2CH2-;

[0544] Xa is a bond or -NR-;

[0545] R is H or -C 1-4 alkyl;

[0546] R1 is selected from H, Boc, NR'R'', and C. 1-4 Alkyl groups, -C(NH)NR'R'', -C(O)NR'R'', and -C(O)OR', preferably selected from H, Boc, and C 1-4 Alkyl groups, -C(NH)NR'R'', -C(O)NR'R'', and -C(O)OR';

[0547] R' and R'' are independently selected from H and -C. 1-4 Alkyl and -C 1-4 alkylene-OH;

[0548] Alternatively, R1 can be selected from Boc, NH2, and C. 1-4 Alkyl groups, -C(NH)NH2, -C(O)NH2, -C(O)NH-C 1-4 Alkyl group, -C(O)NH-C 1-4 Alkylenes -OH, -C(O)OH and -C(O)OC 1-4 alkyl;

[0549] “ "Represents a chiral center, which is selected from the (S) or (R) configuration;

[0550] Alternatively,

[0551] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0552] P3 is selected from: , , , , , , , , , and Alternatively, it can be selected from: , , , , , and ;

[0553] P2 is selected from: , , and ;

[0554] P1 is selected from: , , , , , , , , , , , , and .

[0555] In some embodiments, this disclosure relates to compounds having formula (I) or pharmaceutically acceptable salts thereof, enantiomers, diastereomers, or mixtures thereof, wherein,

[0556] L is represented by L1-L2-X-;

[0557] L1 is the ligand covalently bound part. Alternatively, L1 can form a covalent bond with the functional group of an amino acid, and even more alternatively, L1 can form an amide bond or a thioether bond with the functional group of an amino acid.

[0558] -L2-X- is an extended subunit;

[0559] L2 is selected from C 1-20 Alkylene, -C 2-20 imide and C 2-20 Idemynyl group, in which C 1-20 The 1, 2, 3, 4, 5, 6, 7 or 8 non-adjacent carbon atoms in the alkylene group may optionally be replaced by O or S;

[0560] X either does not exist or is -C(O)-;

[0561] L can be arbitrarily divided by 1, 2, 3, 4 or 5 Rs. L Group substitution;

[0562] R L Selected from H, D, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups.

[0563] Alternatively,

[0564] L is represented by L1-L2-X-;

[0565] L1 is a ligand covalently bonded moiety capable of forming amide or thioether bonds with the functional groups of amino acids, and can alternatively be selected from... and ;

[0566] -L2-X- is an extended subunit;

[0567] L2 is selected from C 1-10 Alkylene, -C 0-4 Alkylene-(C 1-4 alkylene-O) 1-10 -C 0-4 alkylene-, -C 0-4 Alkylene-(OC) 1-4 Alkylene) 1-10 -C 0-4 alkylene-, -C 2-10 imide and C 2-10 Ethyne group;

[0568] X either does not exist or is -C(O)-;

[0569] L can be arbitrarily divided by 1, 2, 3, 4 or 5 Rs. L Group substitution;

[0570] R L Selected from H, D, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0571] Alternatively,

[0572] L is represented by L1-L2-X-;

[0573] L1 is selected from and ;

[0574] L2 is selected from C 1-10 Alkylene, -(CH2) 0-4 -(CH2CH2-O) 1-10 -(CH2) 0-4 -、-(CH2) 0-4 -(O-CH2CH2) 1-10 -(CH2) 0-4 -、C 2-10 imide and C 2-10 Ethyne group;

[0575] X either does not exist or is -C(O)-;

[0576] L can be optionally divided by 1, 2 or 3 Rs L Group substitution;

[0577] R L Selected from H, D, C 1-4 Alkyl and C 1-4 Halogenated alkyl groups;

[0578] Alternatively,

[0579] L is selected from , and ;

[0580] m and n are independently selected from 1, 2, 3, 4, 5, 6, 7 or 8;

[0581] Alternatively, m can be selected from 4, 5, 6, 7 or 8, and can be 4, 5 or 8;

[0582] Alternatively, n can be selected from 2, 3, or 4, and 3 is an alternative.

[0583] In some embodiments, this disclosure relates to compounds having formula (I) or pharmaceutically acceptable salts thereof, enantiomers, diastereomers, or mixtures thereof, wherein,

[0584] Y is selected from: and ;

[0585] R Y1 Selected from H, halogens, OH, CN, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl and C 1-6 Halogenated alkoxy groups;

[0586] R Y2 Selected from H, halogens, OH, CN, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl and C 1-6 Halogenated alkoxy groups;

[0587] p is 0, 1, or 2;

[0588] q is 0, 1, 2, 3, or 4;

[0589] Alternatively,

[0590] Y is selected from: and ;

[0591] R Y1 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0592] R Y2 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0593] p is 0, 1, or 2;

[0594] q is 0, 1, or 2;

[0595] Alternatively,

[0596] Y is selected from: (AM) and (PABC).

[0597] In some embodiments, this disclosure relates to compounds having formula (X) or formula (I), or pharmaceutically acceptable salts, enantiomers, diastereomers, or mixtures thereof:

[0598] Part D of this drug is selected from the following groups: microtubule inhibitors and microtubule polymerization inhibitors, antibiotics, DNA synthesis inhibitors, topoisomerase I inhibitors, immunomodulators, and chelating ligands;

[0599] Alternatively,

[0600] The microtubule inhibitors and microtubule polymerization inhibitors are selected from the group consisting of: olistatins (e.g., MMAE, MMAF, and MMAD), maytansines (e.g., maytansine, maytansinol, DM1, DM2, DM3, and DM4), tubulosyntheticin, candidin, eribulin, and rhizobium.

[0601] This antibiotic was selected from the group consisting of: chalcone, doxorubicin, and anthracyclines;

[0602] The DNA synthesis inhibitor was selected from the group consisting of: betamethasone, PBD (pyrrolobenzodiazepine), and IGN (indolinobenzodiazepine).

[0603] The topoisomerase I inhibitor is selected from the group consisting of: camptothecin and camptothecin analogues (e.g., DXd, SN38 and ixotecan).

[0604] The immunomodulator was selected from the following group: TLR7 agonists, TLR8 agonists, STING agonists, and RIG-I agonists;

[0605] Alternatively,

[0606] D is selected from: , and .

[0607] In some embodiments, this disclosure relates to compounds having formula (I) or pharmaceutically acceptable salts, enantiomers, diastereomers, or mixtures thereof:

[0608] L is represented by L1-L2-X-;

[0609] L1 is a ligand covalently bonded moiety capable of forming amide or thioether bonds with the functional groups of amino acids, and can alternatively be selected from... and ;

[0610] -L2-X- is an extended subunit;

[0611] L2 is selected from C 1-10 Alkylene, -(C 1-4 alkylene-O) 1-10 -、-(OC 1-4 Alkylene) 1-10-、-C 2-10 imide and C 2-10 Ethyne group;

[0612] X either does not exist or is -C(O)-;

[0613] L can be arbitrarily divided by 1, 2, 3, 4 or 5 Rs. L Group substitution;

[0614] R L Selected from H, D, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0615] Y is selected from: and ;

[0616] R Y1 Selected from H, halogens, OH, CN, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl and C 1-6 Halogenated alkoxy groups;

[0617] R Y2 Selected from H, halogens, OH, CN, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl and C 1-6 Halogenated alkoxy groups;

[0618] p is 0, 1, or 2;

[0619] q is 0, 1, 2, 3, or 4;

[0620] Part D of this drug is selected from the following groups: microtubule inhibitors and microtubule polymerization inhibitors, antibiotics, DNA synthesis inhibitors, topoisomerase I inhibitors, immunomodulators, and chelating ligands;

[0621] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0622] Among them, P3 is ,

[0623] Rc1, Rc2, and Rc3 are independently selected from H, OH, CN, NH2, halogens, O-PG, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 3-10 Cycloalkyl, 3- to 10-membered heterocyclic, 5- to 10-membered heteroaryl and C 6-10 Aryl;

[0624] Or Rc2 and Rc3 together with the carbon atoms to which they are attached form C 3-10 Cycloalkylene or 3- to 10-membered heterocyclic cycloidene groups;

[0625] c is 0, 1, 2, 3 or 4;

[0626] P3 may optionally be further substituted with 1, 2, 3, 4 or 5 Rc groups;

[0627] Rc is selected from H, OH, CN, NH2, halogens, PG, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl and C 1-6 The haloalkoxy group can be alternatively selected from H, OH, CN, NH2, halogen, PG, C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0628] P2 is Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl, C 1-6 Halogenated alkyl groups and -(CH2) 1-6 -OH, or Rb1 and Rb2 together with the carbon atoms to which they are attached, form C 3-10 Cycloalkylene or 3- to 10-membered heterocyclic cycloidene groups;

[0629] P2 may optionally be further substituted with 1, 2, 3, 4 or 5 Rb groups;

[0630] Rb is selected from H, halogens, PG, and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0631] P1 is selected from ;

[0632] La is -C 1-8 alkylene-, where -C 1-8 The 1, 2, 3 or 4 non-adjacent carbon atoms in the alkylene group can be optionally replaced by O or S;

[0633] Xa is a bond or -NR-;

[0634] R is H or -C 1-4 alkyl;

[0635] R1 is selected from H, Boc, and C. 1-6 Alkyl groups, -OR', NR'R'', -C(O)R', -C(O)OR', -C(NH)NR'R'', and -C(O)NR'R'', preferably selected from H, Boc, C 1-6Alkyl, -OR', -C(O)R', -C(O)OR', -C(NH)NR'R'' and -C(O)NR'R'';

[0636] R' and R'' are independently selected from H, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, -C 1-6 Alkylene-CN, -C 1-6 Alkyl-NH2 and -C 1-6 alkylene-OH;

[0637] P1 may optionally be further substituted with 1, 2, 3, 4 or 5 Ra groups;

[0638] Ra is selected from H, halogen, PG, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0639] “ "Represents a chiral center, which is selected from the (S) or (R) configuration;

[0640] PG is selected from: trimethylsilyl group (TMS), triethylsilyl group (TES), dimethylisopropylsilyl group (DMIPS), diethylisopropylsilyl group (DEIPS), tert-butyldimethylsilyl group (TBDMS), tert-butyldiphenylsilyl group (TBDPS), triisopropylsilyl group (TIPS), acetyl group (Ac), chloroacetyl group, dichloroacetyl group, trichloroacetyl group, trifluoroacetyl group (TFA), benzoyl group, p-methoxybenzoyl group, 9-fluorenylmethoxycarbonyl group (Fmoc), allyloxycarbonyl group (Alloc). 2,2,2-Trichloroethoxycarbonyl (Troc), carboxybenzyl (Cbz), tert-butoxycarbonyl (Boc), benzyl (Bn), p-methoxybenzyl (PMB), allyl, triphenylmethyl (Tr), bis-p-methoxytriphenylmethyl (DMTr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), p-methoxybenzyloxymethyl (PMBM), -C(O)CH2CH2C(O)OH or 4,4'-dimethoxytriphenylmethyl.

[0641] In some embodiments, this disclosure relates to compounds having formula (I) or pharmaceutically acceptable salts thereof, enantiomers, diastereomers, or mixtures thereof, wherein,

[0642] L is represented by L1-L2-X-;

[0643] L1 is selected from and ;

[0644] L2 is selected from C 1-10 Alkylene, -(CH2) 0-4 -(CH2CH2-O) 1-10 -(CH2) 0-4 -、-(CH2) 0-4 -(O-CH2CH2) 1-10 -(CH2) 0-4 -、C 2-10 imide and C 2-10 Ethyne group;

[0645] X either does not exist or is -C(O)-;

[0646] L can be optionally divided by 1, 2 or 3 Rs L Group substitution;

[0647] R L Selected from H, D, C 1-4 Alkyl and C 1-4 Halogenated alkyl groups;

[0648] Y is selected from: and ;

[0649] R Y1 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0650] R Y2 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0651] Part D of this drug is selected from the following groups: microtubule inhibitors and microtubule polymerization inhibitors, antibiotics, DNA synthesis inhibitors, topoisomerase I inhibitors, immunomodulators, and chelating ligands;

[0652] The microtubule inhibitors and microtubule polymerization inhibitors are selected from the group consisting of: olistatins (e.g., MMAE, MMAF, and MMAD), maytansine (e.g., DM1, DM2, DM3, and DM4), tubulosyntheticin, candidin, eribulin, and rhizobium.

[0653] This antibiotic was selected from the group consisting of: chalcone, doxorubicin, and anthracyclines;

[0654] The DNA synthesis inhibitor was selected from the group consisting of: betamethasone, PBD (pyrrolobenzodiazepine), and IGN (indolinobenzodiazepine).

[0655] The topoisomerase I inhibitor is selected from the group consisting of camptothecin and camptothecin analogues (e.g., DXd, SN38, and ixotecan).

[0656] The immunomodulator was selected from the following group: TLR7 agonists, TLR8 agonists, STING agonists, and RIG-I agonists;

[0657] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0658] Among them, P3 is ,

[0659] Rc1 is selected from OH, O-PG, and C. 1-4 Alkyl groups, Rc2 and Rc3, are independently selected from H, halogens, C. 1-4 Alkyl, 5- to 10-membered heteroaryl (e.g., 5- to 6-membered heteroaryl) and C 6-10 Aryl groups (such as phenyl), or Rc2 and Rc3, together with the carbon atoms to which they are attached, form C 3-5 Cycloalkylene;

[0660] Rc1, Rc2 and Rc3 can be independently and optionally replaced by 1, 2 or 3 Rc;

[0661] Rc is selected from H, OH, C 1-4 Alkyl, C 1-4 Alkoxy and C 1-4 Haloalkyl groups, which may be selected from H, OH, C. 1-4 Alkyl and C 1-4 Halogenated alkyl groups;

[0662] c is 0, 1, or 2;

[0663] P2 is Rb1 and Rb2 are independently selected from H and C. 1-4 Alkyl groups and -(CH2) 1-4 -OH, or Rb1 and Rb2 together with the carbon atoms to which they are attached, form C 3-5 Cycloalkylene;

[0664] P1 is selected from ;

[0665] La is -(CH2) 1-6 -, where -(CH2) 1-6 The 1, 2, or 3 non-adjacent carbon atoms in - can be optionally replaced by O; alternatively, La is selected from -CH2-, -CH2CH2-, CH2CH2CH2-, CH2CH2CH2CH2-, and -CH2OCH2CH2-;

[0666] Xa is a bond or -NR-;

[0667] R is H or -C 1-4 alkyl;

[0668] R1 is selected from H, Boc, NR'R'', and C. 1-4 Alkyl groups, -C(NH)NR'R'', -C(O)NR'R'', and -C(O)OR', preferably selected from H, Boc, and C 1-4 Alkyl groups, -C(NH)NR'R'', -C(O)NR'R'', and -C(O)OR';

[0669] R' and R'' are independently selected from H and -C. 1-4 Alkyl and -C 1-4 alkylene-OH;

[0670] Alternatively, R1 can be selected from Boc, NH2, and C. 1-4 Alkyl groups, -C(NH)NH2, -C(O)NH2, -C(O)NH-C 1-4 Alkyl group, -C(O)NH-C 1-4 Alkylenes -OH, -C(O)OH and -C(O)OC 1-4 alkyl;

[0671] “ "Represents a chiral center, which is selected from the (S) or (R) configuration.

[0672] In some embodiments, this disclosure relates to compounds having formula (I) or pharmaceutically acceptable salts thereof, enantiomers, diastereomers, or mixtures thereof, wherein,

[0673] L is selected from , and ;

[0674] m and n are independently selected from 1, 2, 3, 4, 5, 6, 7 or 8;

[0675] Alternatively, m can be selected from 4, 5, 6, 7 or 8, and can be 4, 5 or 8;

[0676] Alternatively, n can be selected from 2, 3, or 4, and 3 is an alternative.

[0677] Y is selected from: (AM) and (PABC);

[0678] D is selected from: , and ;

[0679] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0680] P3 is selected from: , , , , , , , , , and Alternatively, it can be selected from: , , , , , , and ;

[0681] P2 is selected from: , , and ;

[0682] P1 is selected from: , , , , , , , , , , , , and Preferably selected from , , , , , , , , and .

[0683] In some embodiments, this disclosure relates to compounds having formula (I) or pharmaceutically acceptable salts thereof, enantiomers, diastereomers, or mixtures thereof, wherein,

[0684] L, Y, and D are as defined in the context;

[0685] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0686] -P3-P2-P1- is ,

[0687] “ "Represents a chiral center, which is selected from the (S) or (R) configuration;

[0688] Rc1, Rc2, and Rc3 are independently selected from H, OH, CN, NH2, halogens, and C. 1-6 Alkyl, C 1-6 Alkoxy and C 1-6 Halogenated alkyl groups;

[0689] c is 0, 1, 2, 3 or 4;

[0690] P3 may optionally be further substituted with 1, 2, 3, 4 or 5 Rc groups;

[0691] Rc is selected from H, OH, CN, NH2, halogens, PG, and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0692] Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl, C 1-6 Halogenated alkyl groups and -(CH2) 1-6 -OH;

[0693] P2 may optionally be further substituted with 1, 2, 3, 4 or 5 Rb groups;

[0694] Rb is selected from H, halogens, PG, and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0695] La is -C 1-8 alkylene-, where -C 1-8 The 1, 2, 3 or 4 non-adjacent carbon atoms in the alkylene group can be optionally replaced by O or S;

[0696] Xa is a bond or -NR-;

[0697] R is H or -C 1-4 alkyl;

[0698] R1 is selected from Boc, C 1-6 Alkyl groups, -C(O)R', -C(O)OR', -C(NH)NR'R'' and -C(O)NR'R'';

[0699] R' and R'' are independently selected from H, halogen, C1-6 Alkyl, C 1-6 Halogenated alkyl, -C 1-6 Alkylene-CN, -C 1-6 Alkyl-NH2 and -C 1-6 alkylene-OH;

[0700] P1 may optionally be further substituted with 1, 2, 3, 4 or 5 Ra groups;

[0701] Ra is selected from H, halogen, PG, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0702] alternative

[0703] “ "Represents a chiral center, which is selected from the (S) or (R) configuration;

[0704] Rc1 is selected from H, OH, CN, NH2, halogens, and C. 1-6 Alkyl, C 1-6 Alkoxy and C 1-6 Halogenated alkyl groups;

[0705] Rc2 and Rc3 are independently selected from H and C. 1-6 Alkyl, C 1-6 Alkoxy and C 1-6 Halogenated alkyl groups;

[0706] c is 0, 1, 2, or 3;

[0707] Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0708] La is -C 1-6 alkylene-;

[0709] Xa is -NR-;

[0710] R is H or -C 1-4 alkyl;

[0711] R1 is selected from -C(O)R', -C(O)OR', -C(NH)NR'R'' and -C(O)NR'R'';

[0712] R' and R'' are independently selected from H, halogen, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0713] Alternatively,

[0714] “ "Represents a chiral center, which is selected from the (S) or (R) configuration;

[0715] Rc1 is selected from H, OH, CN, NH2 and halogens, preferably OH;

[0716] Rc2 and Rc3 are independently selected from H and C. 1-6 Alkyl groups, preferably H;

[0717] c is 0, 1, or 2, preferably 0;

[0718] Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl groups, preferably H;

[0719] La is -C 1-6 alkylene-;

[0720] Xa is -NH-;

[0721] R1 is selected from -C(NH)NR'R'' and -C(O)NR'R'', preferably -C(NH)NR'R'';

[0722] R' and R'' are independently selected from H and C. 1-6 Alkyl groups, preferably H;

[0723] Alternatively,

[0724] -P3-P2-P1- is (SGR).

[0725] In some embodiments, this disclosure relates to compounds having formula (I) or pharmaceutically acceptable salts thereof, enantiomers, diastereomers, or mixtures thereof, wherein,

[0726] L, Y, and D are as defined in the context;

[0727] P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y;

[0728] -P3-P2-P1- is ,

[0729] “ "Represents a chiral center, which is selected from the (S) or (R) configuration;

[0730] Rc1, Rc2, and Rc3 are independently selected from H, OH, CN, NH2, halogens, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl groups, 5- to 10-membered heteroaryl groups and C6-10 Aryl;

[0731] c is 0, 1, 2, 3 or 4;

[0732] P3 may optionally be further substituted with 1, 2, 3, 4 or 5 Rc groups;

[0733] Rc is selected from H, OH, CN, NH2, halogens, PG, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl and C 1-6 Halogenated alkoxy groups;

[0734] Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl, C 1-6 Halogenated alkyl groups and -(CH2) 1-6 -OH;

[0735] P2 may optionally be further substituted with 1, 2, 3, 4 or 5 Rb groups;

[0736] Rb is selected from H, halogens, PG, and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0737] La is -C 1-8 alkylene-, where -C 1-8 The 1, 2, 3 or 4 non-adjacent carbon atoms in the alkylene group can be optionally replaced by O or S;

[0738] Xa is a bond or -NR-;

[0739] R is H or -C 1-4 alkyl;

[0740] R1 is selected from H, Boc, and C. 1-6 Alkyl groups, -C(O)R', -C(O)OR', -C(NH)NR'R'' and -C(O)NR'R'';

[0741] R' and R'' are independently selected from H, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, -C 1-6 Alkylene-CN, -C 1-6 Alkyl-NH2 and -C 1-6 alkylene-OH;

[0742] P1 may optionally be further substituted with 1, 2, 3, 4 or 5 Ra groups;

[0743] Ra is selected from H, halogen, PG, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0744] alternative

[0745] “ "Represents a chiral center, which is selected from the (S) or (R) configuration;

[0746] Rc1 and Rc2 are independently selected from H, halogens, OH and C. 1-6 Alkoxy;

[0747] Rc3 is selected from H, halogen, 5- to 10-membered heteroaryl (e.g., 5- to 6-membered heteroaryl), and C. 6-10 Aryl (e.g., phenyl);

[0748] c is 0, 1, 2, or 3;

[0749] Rc1, Rc2 and Rc3 are independently and optionally substituted by 1, 2 or 3 Rc groups;

[0750] Rc is selected from H, OH, CN, NH2, halogens, PG, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl and C 1-6 Halogenated alkoxy groups;

[0751] Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl groups and -(CH2) 1-6 -OH;

[0752] La is -C 1-6 alkylene-;

[0753] Xa is -NR-;

[0754] R is H or -C 1-4 alkyl;

[0755] R1 is selected from -C(O)R', -C(O)OR', -C(NH)NR'R'' and -C(O)NR'R'';

[0756] R' and R'' are independently selected from H, halogen, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups;

[0757] Alternatively,

[0758] “ "Represents a chiral center, which is selected from the (S) or (R) configuration;

[0759] Rc1 and Rc2 are independently selected from H, OH and C. 1-4 Alkyl group, preferably H;

[0760] Rc3 is selected from H, halogen, 5- to 10-membered heteroaryl (e.g., 5- to 6-membered heteroaryl), and C. 6-10 Aryl (e.g., phenyl), preferably phenyl;

[0761] c is 0, 1, or 2, preferably 0;

[0762] Rc1, Rc2 and Rc3 are independently and optionally substituted by 1, 2 or 3 Rc groups;

[0763] Rc is selected from H, OH, C 1-6 Alkyl and C 1-6 Alkoxy;

[0764] Rb1 and Rb2 are independently selected from H and -(CH2). 1-4 -OH, preferably H or -CH2-OH;

[0765] La is -C 1-6 alkylene-;

[0766] Xa is -NH-;

[0767] R1 is selected from -C(NH)NR'R'' and -C(O)NR'R'', preferably -C(NH)NR'R'';

[0768] R' and R'' are independently selected from H and C. 1-6 Alkyl groups, preferably H;

[0769] Alternatively,

[0770] -P3-P2-P1- is (YSR).

[0771] In some embodiments, this disclosure relates to compounds having formula (X) or formula (I) or pharmaceutically acceptable salts, enantiomers, diastereomers, or mixtures thereof, wherein the compound is selected from:

[0772] ;

[0773] ;

[0774] ;

[0775] ;

[0776] ;

[0777] ;

[0778] ;

[0779] ;as well as

[0780] ,

[0781] Where m, n, P3, P2, and P1 are as defined in the context;

[0782] Alternatively,

[0783] m is selected from 4, 5, 6, 7 or 8, and can be 4, 5 or 8;

[0784] n is selected from 2, 3 or 4, and can be 3;

[0785] In some embodiments, this disclosure relates to a compound having formula (X) or formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MsP-SGR-DXD:

[0786] .

[0787] In some embodiments, this disclosure relates to a compound having formula (X) or formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MsP-YSR-DXD:

[0788] .

[0789] In some embodiments, this disclosure relates to a compound having formula (X) or formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MsP-SGR-ecientacan:

[0790] .

[0791] In some embodiments, this disclosure relates to a compound having formula (X) or formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MsP-YSR-ecientacan:

[0792] .

[0793] In some embodiments, this disclosure relates to a compound having formula (X) or formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein the compound is MsP-SGR-PABC-Ecinotecan:

[0794] .

[0795] In some embodiments, this disclosure relates to a compound having formula (X) or formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein the compound is MsP-YSR-PABC-Ecinotecan:

[0796] .

[0797] In some embodiments, this disclosure relates to a compound having formula (X) or formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MsP-SGE-DXD:

[0798] .

[0799] In some embodiments, this disclosure relates to a compound having formula (X) or formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MsP-TSR-DXD:

[0800] .

[0801] In some embodiments, this disclosure relates to a compound having formula (X) or formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer thereof, or a mixture thereof, wherein the compound is MsP-SGK-DXD:

[0802] .

[0803] In some embodiments, this disclosure relates to a compound having formula (X) or formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein the compound is MsP-Y(Me)SR-DXD:

[0804] .

[0805] In some embodiments, this disclosure relates to a compound having formula (X) or formula (I) or a pharmaceutically acceptable salt thereof, an enantiomer, a diastereomer, or a mixture thereof, wherein the compound is Mal-PEG8-SGR-PABC-Ecinotecan:

[0806] .

[0807] In some embodiments, this disclosure relates to compounds having formula (X) or formula (I), or pharmaceutically acceptable salts, enantiomers, diastereomers, or mixtures thereof, wherein,

[0808] The drug is derived from compounds having the formula (DI) or (D-II):

[0809] (DI) or (D-II)

[0810] Rd1 is C 1-6 Alkyl or C 1-6 Alkoxy;

[0811] Rd2 is a halogen;

[0812] Or preferably, Rd1 and Rd2 together with the carbon atoms to which they are attached form a 5-10 membered heterocyclic group or a 5-10 membered heteroarylene group, which is optionally surrounded by one, two or three atoms selected from H, D, halogens and C. 1-6 Alkyl substituents;

[0813] Rd3 is selected from -(CH2). p -ORd a -(CH2) p -C(O)ORd a -(CH2) p -NHC(O)-Rd a and -(CH2) p -CH2NRd b Rd c ;

[0814] Rd3 is preferably selected from -(CH2). p -ORd a -(CH2) p -C(O)ORd a and -(CH2) p -NHC(O)-Rd a ;

[0815] Rd a Selected from H, C 1-6 Alkyl, C 1-6 alkylene-OH, C 1-6 Alkylene-NH2, such as H, CH3, CH2OH, CH2CH2OH and CH2CH2NH2;

[0816] Rd b and Rd c Independently selected from H and C 1-6 Alkyl groups, such as H and CH3;

[0817] Each p is independently 0, 1, 2, 3, 4, 5, or 6;

[0818] Alternatively,

[0819] Rd1 is C 1-6 Alkyl or C 1-6 Alkoxy;

[0820] Rd2 is a halogen;

[0821] Or preferably, Rd1 and Rd2 together with the carbon atoms to which they are attached form a 5-6 membered heterocyclic cycloidene or a 5-6 membered heteroarylene (preferably forming a 5-6 membered heterocyclic cycloidene), which is optionally surrounded by one, two or three atoms selected from H, D, halogens and C. 1-4 Alkyl substituents;

[0822] Rd3 is selected from , , , , or Preferably ;

[0823] Rd a 、Rd b and Rd c Independently selected from H and C 1-6 alkyl;

[0824] Each p is independently 0, 1, 2, 3, 4, 5, or 6;

[0825] Alternatively,

[0826] Rd1 is C 1-4 Alkyl or C 1-4 Alkyl groups, such as Me or OMe;

[0827] Rd2 is a halogen, such as F;

[0828] Or preferably, Rd1, Rd2 together with the carbon atoms to which they are attached form , or ;

[0829] Rd3 is selected from , , , , , , , , , and ;

[0830] Alternatively, part D of the drug is derived from the following compounds:

[0831] , , , , , , , , , , , or .

[0832] In some embodiments, this disclosure relates to compounds having formula (X) or formula (I), or pharmaceutically acceptable salts, enantiomers, diastereomers, or mixtures thereof, wherein the pharmaceutical portion D is selected from compounds having formulas (D-III) or (D-IV):

[0833] (D-III) (D-IV)

[0834] in,

[0835] Indicates the attachment site with the rest of the compound;

[0836] Rd1 and Rd2 are as defined in the context;

[0837] Rd3' is selected from -(CH2) p -ORd a '-、-(CH2) p -C(O)ORd a '-、-(CH2) p -NHC(O)-Rd a '- and -(CH2) p -CH2NRd b Rd c '-;

[0838] Rd3' is preferably selected from -(CH2). p -ORd a '-、-(CH2) p -C(O)ORd a '- and -(CH2) p -NHC(O)Rd a '-;

[0839] Rd a 'Selected from key, C 1-6 Alkylene, -C 1-6 alkylene-O- and -C 1-6Alkylene -NH-, such as bond, -CH2-, -CH2O-, -CH2CH2O- and -CH2CH2NH-;

[0840] Rd b Selected from H and C 1-6 Alkyl groups, such as H and CH3;

[0841] Rd c 'Selected from key and C' 1-6 Alkyl groups, such as bonds and -CH2-;

[0842] Each p is independently 0, 1, 2, 3, 4, 5, or 6;

[0843] Alternatively,

[0844] Rd3' is selected from , , , , and ,For example , , , , , , , , , and Preferably ;

[0845] Rd a 'and Rd c 'Independently selected from key and C 1-6 Alkylene;

[0846] Rd b Selected from H and C 1-6 alkyl;

[0847] Each p is independently 0, 1, 2, 3, 4, 5, or 6.

[0848] Alternatively,

[0849] Part D of this drug is selected from:

[0850] , , , , , , , , , , , or .

[0851] In some embodiments, this disclosure relates to compounds having formula (X) or formula (I) or pharmaceutically acceptable salts, enantiomers, diastereomers, or mixtures thereof, wherein the compound is selected from:

[0852] , ,

[0853] , ,

[0854] , ,

[0855] Where m, n, P3, P2, and P1 are as defined in the context;

[0856] Alternatively,

[0857] m is selected from 4, 5, 6, 7 or 8, and can be 4, 5 or 8;

[0858] n is selected from 2, 3 or 4, and can be 3;

[0859] Alternatively, the compound may be selected from:

[0860] ;

[0861] .

[0862] II. Couplings

[0863] In one aspect of this disclosure, a coupling having formula (II) is provided:

[0864] T-(S'-D)k (II)

[0865] in,

[0866] T stands for the target region;

[0867] S' is a linker, which is a divalent group formed by linking S and T;

[0868] The linker contains a peptide-cleavable unit that contains a tripeptide having the sequence -P3-P2-P1-, where P3, P2, and P1 are as defined in the context.

[0869] D represents the drug component;

[0870] k ranges from 1 to approximately 20.

[0871] In some embodiments, the targeting portion is selected from an antibody or its antigen-binding fragment, ligand, or targeting peptide.

[0872] In some embodiments, the antibody or its antigen-binding fragment binds to a tumor antigen, a tumor-associated antigen, or an immune cell antigen or a T cell antigen;

[0873] The ligand binds to a receptor expressed on tumor cells or immune cells, or to a receptor expressed in the tumor microenvironment; and / or;

[0874] The targeting peptide binds to target molecules expressed on tumor cells or immune cells, or to receptors expressed in the tumor microenvironment.

[0875] In some embodiments, the antibody or its antigen-binding fragment, ligand or targeting peptide binds to a protein, including but not limited to CD163, CD74, CXCR4, CD11, CD71, CD70, TNFRSF13c, CD30, IL-6, FRβ and TNFβ.

[0876] In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a bispecific antibody. In some embodiments, the antibody is a multispecific antibody.

[0877] In some embodiments, the antibody is a mouse antibody. In some embodiments, the antibody is a chimeric antibody. In some embodiments, the antibody is a humanized antibody. In some embodiments, the antibody is a human antibody.

[0878] In some embodiments, the antibody belongs to an isotype selected from the group consisting of IgG, IgA, IgM, IgE, and IgD. In some embodiments, the antibody belongs to a subtype selected from the group consisting of IgG1, IgG2, IgG3, and IgG4.

[0879] In some embodiments, the antigen-binding fragment is selected from the group consisting of Fab, Fab', F(ab')2, Fv, scFv and ds-scFv.

[0880] In some embodiments, k ranges from 1 to about 20; in other embodiments, k ranges from 1 to about 10; in other embodiments, k ranges from 2 to about 9; in other embodiments, k ranges from 4 to about 8; in other embodiments, k is about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or about 10.

[0881] In some embodiments, -S'-D is represented by equation (III):

[0882] (III)

[0883] in Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0884] L' is represented by the formula -L1'-L2-X-, where L1' is a divalent group formed by connecting L1 and T;

[0885] L1, L2, P, Y, and D are as defined in the context;

[0886] Alternatively, L1' is selected from , , and Alternatively, L1' is selected from and .

[0887] In some embodiments, -S'-D is:

[0888] ,

[0889] in, Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0890] m, P3, P2, and P1 are as defined in the context.

[0891] In some embodiments, -S'-D is:

[0892] ,

[0893] in, Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0894] m, P3, P2, and P1 are as defined in the context.

[0895] In some embodiments, -S'-D is:

[0896] ,

[0897] in, Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0898] m, P3, P2, and P1 are as defined in the context.

[0899] In some embodiments, -S'-D is:

[0900] ,

[0901] in, Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0902] m, P3, P2, and P1 are as defined in the context.

[0903] In some embodiments, -S'-D is:

[0904] ,

[0905] in, Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0906] m, P3, P2, and P1 are as defined in the context.

[0907] In some embodiments, -S'-D is:

[0908] ,

[0909] in, Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0910] n, P3, P2, and P1 are defined as in the context.

[0911] In some embodiments, -S'-D is:

[0912] ,

[0913] in, Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0914] n, P3, P2, and P1 are defined as in the context.

[0915] In some embodiments, -S'-D is:

[0916] ,

[0917] in, Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0918] n, P3, P2, and P1 are defined as in the context.

[0919] In some embodiments, -S'-D is:

[0920] ,

[0921] in, Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0922] n, P3, P2, and P1 are defined as in the context.

[0923] In some embodiments, -S'-D is:

[0924] ,

[0925] in, Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0926] m, P3, P2, and P1 are as defined in the context.

[0927] In some embodiments, -S'-D is:

[0928] ,

[0929] in, Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0930] n, P3, P2, and P1 are defined as in the context.

[0931] In some embodiments, -S'-D is:

[0932] ,

[0933] in, Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0934] n, P3, P2, and P1 are defined as in the context.

[0935] In some embodiments, -S'-D is:

[0936] ,

[0937] in, Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0938] n, P3, P2, and P1 are defined as in the context.

[0939] In some embodiments, -S'-D is:

[0940] ,

[0941] in, Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0942] n, P3, P2, and P1 are defined as in the context.

[0943] In some embodiments, -S'-D is:

[0944] ,

[0945] in, Indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context;

[0946] n, P3, P2, and P1 are defined as in the context.

[0947] In some specific embodiments, -S'-D is -MC-VGR-DXD:

[0948] ,

[0949] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0950] In some specific embodiments, -S'-D is -MC-SGR-DXD:

[0951] ,

[0952] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0953] In some specific embodiments, -S'-D is -MC-VGR-PABC-MMAE:

[0954] ,

[0955] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0956] In some specific embodiments, -S'-D is -MC-SGR-PABC-MMAE:

[0957] ,

[0958] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0959] In some specific embodiments, -S'-D is -MC-SGC-DXD:

[0960] ,

[0961] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0962] In some specific embodiments, -S'-D is -MC-SAC-DXD:

[0963] ,

[0964] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0965] In some specific embodiments, -S'-D is -MsP-SGC-DXD:

[0966] ,

[0967] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0968] In some specific embodiments, -S'-D is -MC-S(tBu)GC-DXD:

[0969] ,

[0970] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0971] In some specific embodiments, -S'-D is -MC-SGOrn(Boc)-DXD:

[0972] ,

[0973] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0974] In some specific embodiments, -S'-D is -MC-SGC-PABC-Eciticon:

[0975] ,

[0976] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0977] In some specific embodiments, -S'-D is -MC-SGO(DEt)-PABC-Eciticon:

[0978] ,

[0979] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0980] In some specific embodiments, -S'-D is -MC-SGE-PABC-Eciticon:

[0981] ,

[0982] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0983] In some specific embodiments, -S'-D is -MC-SGE(tBu)-PABC-Eciticon:

[0984] ,

[0985] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0986] In some specific embodiments, -S'-D is -MC-SGC(Me)-PABC-Ecitidine:

[0987] ,

[0988] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0989] In some specific embodiments, -S'-D is -MC-SGC(Peg)-PABC-Ecitidine:

[0990] ,

[0991] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0992] In some specific embodiments, -S'-D is -MC-TGC-PABC-Eciticon:

[0993] ,

[0994] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0995] In some specific embodiments, -S'-D is -MC-SGhC(O)-PABC-Ecitidine:

[0996] ,

[0997] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[0998] In some specific embodiments, -S'-D is -MC-YSR-PABC-Ecitidine:

[0999] ,

[1000] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1001] In some specific embodiments, -S'-D is -MC-SGE-DXD:

[1002] ,

[1003] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1004] In some specific embodiments, -S'-D is -MC-SGC(Peg)-DXD:

[1005] ,

[1006] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1007] In some specific embodiments, -S'-D is -MC-TSR-DXD:

[1008] ,

[1009] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1010] In some specific embodiments, -S'-D is -MC-YSR-DXD:

[1011] ,

[1012] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1013] In some specific embodiments, -S'-D is -MC-SGO-DXD:

[1014] ,

[1015] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1016] In some specific embodiments, -S'-D is -MC-SGK-DXD:

[1017] ,

[1018] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1019] In some specific embodiments, -S'-D is -Mal-PEG4-VGR-DXD:

[1020] ,

[1021] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1022] In some specific embodiments, -S'-D is -Mal-PEG4-SGR-DXD:

[1023] ,

[1024] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1025] In some specific embodiments, -S'-D is -MC-SGhR-DXD:

[1026] ,

[1027] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1028] In some specific embodiments, -S'-D is -MC-SGN-DXD:

[1029] ,

[1030] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1031] In some specific embodiments, -S'-D is -MsP-SGR-DXD:

[1032] ,

[1033] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1034] In some specific embodiments, -S'-D is -MsP-YSR-DXD:

[1035] ,

[1036] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1037] In some specific embodiments, -S'-D is -MsP-SGR-Ecinotecan:

[1038] ,

[1039] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1040] In some specific embodiments, -S'-D is -MsP-YSR-Ecinotecan:

[1041] ,

[1042] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1043] In some specific embodiments, -S'-D is -MsP-SGR-PABC-Ecinotecan:

[1044] ,

[1045] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1046] In some specific embodiments, -S'-D is -MsP-YSR-PABC-Ecinotecan:

[1047] ,

[1048] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1049] In some specific embodiments, -S'-D is -MsP-SGE-DXD:

[1050] ,

[1051] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1052] In some specific embodiments, -S'-D is -MsP-TSR-DXD:

[1053] ,

[1054] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1055] In some specific embodiments, -S'-D is -MsP-SGK-DXD:

[1056] ,

[1057] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1058] In some specific embodiments, -S'-D is -MsP-Y(Me)SR-DXD:

[1059] ,

[1060] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1061] In some specific embodiments, -S'-D is -Mal-PEG8-SGR-PABC-Ecinotecan:

[1062] ,

[1063] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1064] In some specific embodiments, -S'-D is:

[1065] ,

[1066] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1067] In some specific embodiments, -S'-D is:

[1068] .

[1069] in, This indicates the attachment site for an antibody or its antigen-binding fragment as defined in the context.

[1070] In some specific embodiments, the coupling element is:

[1071] ,

[1072] Wherein, Ab is an antibody or its antigen-binding fragment.

[1073] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1074] In some specific embodiments, the coupling element is:

[1075] ,

[1076] Wherein, Ab is an antibody or its antigen-binding fragment.

[1077] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1078] In some specific embodiments, the coupling element is:

[1079] ,

[1080] Wherein, Ab is an antibody or its antigen-binding fragment.

[1081] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1082] In some specific embodiments, the coupling element is:

[1083] ,

[1084] Wherein, Ab is an antibody or its antigen-binding fragment.

[1085] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1086] In some specific embodiments, the coupling element is:

[1087] ,

[1088] Wherein, Ab is an antibody or its antigen-binding fragment.

[1089] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1090] In some specific embodiments, the coupling element is:

[1091] ,

[1092] Wherein, Ab is an antibody or its antigen-binding fragment.

[1093] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1094] In some specific embodiments, the coupling element is:

[1095] ,

[1096] Wherein, Ab is an antibody or its antigen-binding fragment.

[1097] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1098] In some specific embodiments, the coupling element is:

[1099] ,

[1100] Wherein, Ab is an antibody or its antigen-binding fragment.

[1101] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1102] In some specific embodiments, the coupling element is:

[1103] ,

[1104] Wherein, Ab is an antibody or its antigen-binding fragment.

[1105] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1106] In some specific embodiments, the coupling element is:

[1107] ,

[1108] Wherein, Ab is an antibody or its antigen-binding fragment.

[1109] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1110] In some specific embodiments, the coupling element is:

[1111] ,

[1112] Wherein, Ab is an antibody or its antigen-binding fragment.

[1113] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1114] In some specific embodiments, the coupling element is:

[1115] ,

[1116] Wherein, Ab is an antibody or its antigen-binding fragment.

[1117] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1118] In some specific embodiments, the coupling element is:

[1119] ,

[1120] Wherein, Ab is an antibody or its antigen-binding fragment.

[1121] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1122] In some specific embodiments, the coupling element is:

[1123] ,

[1124] Wherein, Ab is an antibody or its antigen-binding fragment.

[1125] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1126] In some specific embodiments, the coupling element is:

[1127] ,

[1128] Wherein, Ab is an antibody or its antigen-binding fragment.

[1129] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1130] In some specific embodiments, the coupling element is:

[1131] ,

[1132] Wherein, Ab is an antibody or its antigen-binding fragment.

[1133] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1134] In some specific embodiments, the coupling element is:

[1135] ,

[1136] Wherein, Ab is an antibody or its antigen-binding fragment.

[1137] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1138] In some specific embodiments, the coupling element is:

[1139] ,

[1140] Wherein, Ab is an antibody or its antigen-binding fragment.

[1141] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1142] In some specific embodiments, the coupling element is:

[1143] ,

[1144] Wherein, Ab is an antibody or its antigen-binding fragment.

[1145] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1146] In some specific embodiments, the coupling element is:

[1147] ,

[1148] Wherein, Ab is an antibody or its antigen-binding fragment.

[1149] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1150] In some specific embodiments, the coupling element is:

[1151] ,

[1152] Wherein, Ab is an antibody or its antigen-binding fragment.

[1153] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1154] In some specific embodiments, the coupling element is:

[1155] ,

[1156] Wherein, Ab is an antibody or its antigen-binding fragment.

[1157] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1158] In some specific embodiments, the coupling element is:

[1159] ,

[1160] Wherein, Ab is an antibody or its antigen-binding fragment.

[1161] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1162] In some specific embodiments, the coupling element is:

[1163] ,

[1164] Wherein, Ab is an antibody or its antigen-binding fragment.

[1165] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1166] In some specific embodiments, the coupling element is:

[1167] ,

[1168] Wherein, Ab is an antibody or its antigen-binding fragment.

[1169] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1170] In some specific embodiments, the coupling element is:

[1171] ,

[1172] Wherein, Ab is an antibody or its antigen-binding fragment.

[1173] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1174] In some specific embodiments, the coupling element is:

[1175] ,

[1176] In some specific embodiments, the coupling element is:

[1177] ,

[1178] Wherein, Ab is an antibody or its antigen-binding fragment.

[1179] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1180] In some specific embodiments, the coupling element is:

[1181] ,

[1182] Wherein, Ab is an antibody or its antigen-binding fragment.

[1183] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1184] In some specific embodiments, the coupling element is:

[1185] ,

[1186] Wherein, Ab is an antibody or its antigen-binding fragment.

[1187] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1188] In some specific embodiments, the coupling element is:

[1189] ,

[1190] Wherein, Ab is an antibody or its antigen-binding fragment.

[1191] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1192] In some specific embodiments, the coupling element is:

[1193] ,

[1194] Wherein, Ab is an antibody or its antigen-binding fragment.

[1195] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1196] In some specific embodiments, the coupling element is:

[1197] ,

[1198] Wherein, Ab is an antibody or its antigen-binding fragment.

[1199] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1200] In some specific embodiments, the coupling element is:

[1201] ,

[1202] Wherein, Ab is an antibody or its antigen-binding fragment.

[1203] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1204] In some specific embodiments, the coupling element is:

[1205] ,

[1206] Wherein, Ab is an antibody or its antigen-binding fragment.

[1207] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1208] In some specific embodiments, the coupling element is:

[1209] ,

[1210] Wherein, Ab is an antibody or its antigen-binding fragment.

[1211] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1212] In some specific embodiments, the coupling element is:

[1213] ,

[1214] Wherein, Ab is an antibody or its antigen-binding fragment.

[1215] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1216] In some specific embodiments, the coupling element is:

[1217] ,

[1218] Wherein, Ab is an antibody or its antigen-binding fragment.

[1219] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1220] In some specific embodiments, the coupling element is:

[1221] ,

[1222] Wherein, Ab is an antibody or its antigen-binding fragment.

[1223] k is an integer selected from 1 to 10, preferably 4 to 8, and more preferably k is 4 or 8.

[1224] In some specific embodiments, the coupling element is:

[1225] ,

[1226] Among them, Ab is an antibody or its antigen-binding fragment that binds to tumor antigens, tumor-associated antigens, or immune cell antigens or T cell antigens;

[1227] And k is an integer selected from 1 to 10, preferably 4 to 8, more preferably k is 4 or 8.

[1228] In some specific embodiments, the coupling element is:

[1229] ,

[1230] Among them, Ab is an antibody or its antigen-binding fragment that binds to tumor antigens, tumor-associated antigens, or immune cell antigens or T cell antigens;

[1231] And k is an integer selected from 1 to 10, preferably 4 to 8, more preferably k is 4 or 8.

[1232] In some embodiments of any of the above conjugates, Ab is an antibody or antigen-binding fragment thereof that binds to a tumor antigen, tumor-associated antigen, or immune cell antigen or T cell antigen.

[1233] In some embodiments of any of the above conjugates, Ab is an antibody or antigen-binding fragment thereof that binds to a tumor-associated antigen (TAA); the tumor-associated antigen (TAA) includes, but is not limited to, HER2, TROP2, EGFR, FLOR2, MUC16, CEACAM, nectin4, CD19, CD33, MSLN, EpCAM, NaPi2b, ROR1, CLDN6, CLDN18.2, ROR2, LY6G6D, fibronectin, B7H3, B7H4, BCMA, TF, MET, and others.

[1234] In some embodiments of any of the above conjugates, Ab is a TROP2 antibody.

[1235] In some embodiments, the Trop2 antibody comprises a heavy chain (HC) and a light chain (LC), wherein:

[1236] The HC contains an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity with SEQ ID NO: 1, and the LC contains an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity with SEQ ID NO: 2.

[1237] III. Antibodies

[1238] As used herein, the term "antibody" refers to an immunoglobulin molecule that has the ability to specifically bind to an antigen. Antibodies typically include a variable region and a constant region in each of their heavy and light chains. The variable regions of both the heavy and light chains contain binding domains that interact with the antigen. The constant regions of an antibody mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (such as effector cells) and components of the complement system (such as C1q, the first component in the classical pathway of complement activation). Most antibodies have a heavy chain variable region (VH) and a light chain variable region (VL), which together form the antigen-binding portion of the antibody.

[1239] The light chain variable region (VL) or heavy chain variable region (VH) consists of four frame regions separated by three complementarity-determining regions (CDRs). The frame regions align the CDRs to specifically bind to the epitopes of the antigen. The CDRs contain the amino acid residues of the antibody primarily responsible for antigen binding. From the amino terminus to the carboxyl terminus, both the VL and VH domains contain the following frame (FR) regions and CDRs: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. CDRs 1, 2, and 3 of the VL domain are also referred to herein as LCDR1, LCDR2, and LCDR3, respectively; and CDRs 1, 2, and 3 of the VH domain are also referred to herein as HCDR1, HCDR2, and HCDR3, respectively.

[1240] As used herein, the term "antibody" should be understood in its broadest sense and includes monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, antibody fragments, and multispecific antibodies (e.g., bispecific antibodies) containing at least two distinct antigen-binding regions. Antibodies may contain additional modifications, such as non-naturally occurring amino acids, mutations in the Fc region, and mutations in glycosylation sites. Antibodies also include post-translational modified antibodies, fusion proteins containing antibody antigenic determinants, and any other modified immunoglobulin molecules containing antigen recognition sites, provided that these antibodies exhibit the desired biological activity.

[1241] As used herein, the term "antigen-binding fragment" of an antibody refers to one or more fragments of an antibody that retain the ability to bind specifically to an antigen. It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody.

[1242] Examples of antigen-binding fragments of antibodies include (i) Fab fragments, monovalent fragments consisting of VL, VH, CL, and CH1 domains; (ii) F(ab')2 fragments, bivalent fragments comprising two Fab fragments linked by disulfide bonds through a hinge region; (iii) Fab' fragments, which are essentially Fab fragments with a partially hinge region (see Fundamental Immunology (Paul, ed., 3.sup.rd ed. 1993)); (iv) Fd fragments consisting of VH and CH1 domains; (v) Fd' fragments having VH and CH1 domains and one or more cysteine ​​residues at the C-terminus of the CH1 domain; (vi) Fv fragments consisting of the VL and VH domains of a single antibody arm; and (vii) dAb fragments consisting of the VH domain (Ward et al., (1989) Nature). (viii) Separate complementarity-determining regions (CDRs); and (ix) nanobodies comprising a heavy chain variable region containing a single variable domain and two constant domains. Furthermore, although the two domains VL and VH of the Fv fragment are encoded by separate genes, they can be linked by synthetic linkers using recombinant methods, enabling them to be made into a single protein chain in which the VL and VH regions pair to form a monovalent molecule (called a single-chain Fv (scFv); see, for example, Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). Such single-chain antibodies are also intended to be encompassed in the antibody terminology of “antigen-binding fragment”. In addition, the term also includes “linear antibodies” comprising a pair of tandem Fd fragments (VH-CH1-VH-CH1) that together with a complementary light chain polypeptide form an antigen-binding region, as well as modified forms of any of the aforementioned fragments that retain antigen-binding activity.

[1243] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous population of antibodies. That is, each antibody that makes up the population is identical, except for a small number that may mutate naturally. Monoclonal antibodies are highly specific and target a single antigen. The term "monoclonal antibody" as used herein is not limited to antibodies produced by hybridoma technology and should not be construed as requiring antibodies to be produced by any particular method.

[1244] In the context of this invention, the term "bispecific antibody" is understood to refer to an antibody having two distinct antigen-binding regions defined by different antibody sequences. This can be understood as binding to different targets, but also includes binding to different epitopes within a single target.

[1245] IV. Connector

[1246] The terms “linker” and “drug linker” are used interchangeably in this article and refer to the chemical or biological part that links an antibody to a cytotoxic payload.

[1247] One or more drug moieties can be indirectly coupled to an antibody (e.g., via a linker having direct covalent or non-covalent interactions). The linker can be a chemical linker, such as homo- and hetero-bifunctional crosslinkers available from many commercial sources.

[1248] In some embodiments, the linker S further comprises a cleavable linker or an uncleavable linker; in some embodiments, the cleavable linker comprises an acid-instable linker, a hydrophilic linker, a protease-sensitive linker, a light-instable linker, a hydrazone linker, a dimethyl linker, or a disulfide-containing linker. In some embodiments, the linker S is derived from the following groups: sulfonyl-SPDB (N-succinimide-4-(2-pyridyldithio)-2-sulfonyl-butyrate), SPDB, MC (6-maleimide-hexanoyl), dimethylethylamine (DMEA), CL2A, Mal-PEG8, MsP, AcBut (4-(4-acetylphenoxy)-butyric acid), dibenzocyclooctylene (DBCO), bicyclo[6.1.0]nonyne (BCN), BCN-PEG3-, dimethylhydrazine (3-methyl-3-mercaptobutanehydrazine), AcBut-dimethylhydrazine, or SMCC (N-succinimide-4-(N-maleimide-methyl)cyclohexane-formate).

[1249] In some embodiments, the linker S further comprises MC (6-maleimide hexanoyl): In some embodiments, the connector S further includes MsP: In some embodiments, the linker S further comprises sulfonyl-SPDB (N-succinimide-4-(2-pyridyldithio)-2-sulfonyl-butyrate): In some embodiments, the connection sub-S further includes an SPDB: In some embodiments, linker S further comprises CL2A, which is an incleavable, complex PABC-peptide-mc linker containing PEG8 and triazole; in some embodiments, linker S further comprises dibenzocyclooctylene (DBCO). In some embodiments, the connector S further comprises bicyclic [6.1.0]nonyne (BCN): In some embodiments, the linker S further comprises AcBut-dimethylhydrazine: In some embodiments, the linker S further comprises SMCC (N-succinimide-4-(N-maleimide-methyl)cyclohexane-formate): .

[1250] In some embodiments, the linker S is derived from or includes one or more of the following groups: C 1-10 Alkyl, C 1-10 alkenyl, C 1-10 alkynyl group, C 1-10 Alkylene, C 2-10 imidene group, C 2-10 Ethyne group, (OC) 1-10 Alkylene) 1-10 -O-, -S-, -SH, -N3, -C(O)-, -C(O)NH-, -OC(O)NH-, cycloalkyl, heteroaryl, aryl, heteroaryl, cycloalkylene, heteroarylene, arylene, heteroarylene , , , , , (BCN) (BCN-PEG3) (DBCO) , , , , , , , , , , , , , , , , , , , and their combinations,

[1251] Among them, R s It is hydrogen, halogen, OR, -NO2, -CN, -S(O)2R, C 1-24 Alkyl or 6-24 aryl or heteroaryl, wherein C 1-24 Alkyl or 6-24 aryl or heteroaryl groups are optionally substituted with one or more aryl or heteroaryl groups; or two R groups are substituted with R groups.s Together they form 6-24 membered cycloalkyl, heterocycloalkyl, aryl, or heteroaryl rings;

[1252] R S 'Is C(R) s )2, O, S or NR; and

[1253] R is hydrogen, alkyl, heteroalkyl, cycloalkyl, or heterocycloalkyl;

[1254] s1 is an integer 1, 2, 3, 4, 5 or 6;

[1255] s2 is an integer 1, 2, 3, 4, 5 or 6.

[1256] In some embodiments, the linker (S) in this disclosure includes a ligand covalently binding portion (L1).

[1257] In some embodiments, L1 can form a covalent bond with the functional group of an amino acid. In some embodiments, L1 can form an amide bond or a thioether bond with the functional group of an amino acid. In some particular embodiments, the ligand covalently bound portion (L1) is In other specific embodiments, the ligand covalently binding portion (L1) is In other specific embodiments, the ligand covalently binding portion (L1) is In other specific embodiments, the ligand covalently binding portion (L1) is .

[1258] After Ab is connected to the linker, L1' serves as the connecting portion of the linker and is a divalent group of L1. In some embodiments, L1' is... In other embodiments, L1' is .

[1259] The connector in this disclosure includes an extension subunit (-L2-X-). In some embodiments, L2 is selected from C. 1-10 Alkylene, -C 2-10 imide and C 2-10 Idemynyl group, in which C 1-10 The 1, 2, 3, 4 or 5 non-adjacent carbon atoms in the alkylene group may optionally be replaced by O or S; X is absent or -C(O)-;

[1260] In some embodiments, the connectors in this disclosure include In other embodiments, the connector in this disclosure includes In other embodiments, the connector in this disclosure includes .

[1261] The linker (S) comprises a peptide-cleavable unit having a tripeptide having the sequence -P3-P2-P1-. In some embodiments, the linker (S) further comprises a cleavable linker or an uncleavable linker. In some embodiments, the cleavable linker comprises an acid-labile linker, a hydrophilic linker, a protease-sensitive linker, a light-labile linker, a hydrazone linker, a dimethyl linker, or a disulfide-containing linker.

[1262] The linker in this disclosure comprises a peptide-cleavable unit (P). In some embodiments, P contains a tripeptide directly attached to the drug moiety, and therefore Y is absent. When Y is present and is a self-destructive spacer unit, the tripeptide is attached to the self-destructive spacer unit and thereby cleaved by a protease, providing a drug linker fragment of formula YD, wherein Y undergoes self-destruction to completely release the free drug.

[1263] The peptide cleavable unit comprises a tripeptide having the sequence -P3-P2-P1-, where P3 is linked to X, P1 is linked to Y, and P1, P2, and P3 are amino acid residues that impart selectivity to protease cleavage of tumor tissue homogenates compared to normal tissue homogenates as described herein.

[1264] In some embodiments, P3 is selected from serine, tyrosine, or analogues thereof, wherein a serine analogue refers to a chemically or biologically modified serine, and a tyrosine analogue refers to a chemically or biologically modified tyrosine. In some embodiments, P3 is selected from serine or analogues thereof, wherein a serine analogue refers to a chemically or biologically modified serine.

[1265] In some embodiments, P3 is or It may optionally be further substituted with 1, 2, 3, 4 or 5 Rc groups, wherein these variables are defined as in the context.

[1266] In some specific embodiments, P3 is selected from: , , , , , , , , , and .

[1267] In some specific embodiments, P3 is selected from: , , , , , , and .

[1268] In some embodiments, P2 is selected from glycine, serine, or analogues thereof, wherein a glycine analogue refers to chemically or biologically modified glycine, and a serine analogue refers to chemically or biologically modified serine. In some embodiments, P2 is selected from glycine or analogues thereof, wherein a glycine analogue refers to chemically or biologically modified glycine.

[1269] In some embodiments, P2 is or It may optionally be further replaced by 1, 2, 3, 4 or 5 Rb groups, wherein these variables are defined in the context.

[1270] In some specific embodiments, P2 is selected from: , , and .

[1271] In some embodiments, P1 is selected from citrulline, arginine, glutamic acid, or analogues thereof, wherein an amino acid analogue refers to a chemically or biologically modified amino acid.

[1272] In some embodiments, P1 is or It may optionally be further replaced by 1, 2, 3, 4 or 5 Ra groups, wherein these variables are defined as in the context.

[1273] In some specific embodiments, P1 is selected from: , , , , , , , , , , , , and .

[1274] The linkers in this disclosure comprise spacer units (Y). Each spacer unit may comprise one or more self-destructive spacer units. As used herein, the term "self-destructive spacer" refers to a bifunctional chemical moiety capable of covalently linking two spacer portions (e.g., a coagulation factor or procoagulant peptide and a protein-cleavable substrate) together to form a normally stable tripartite molecule. If the bond between the self-destructive spacer and the first portion (e.g., the protein-cleavable substrate) is cleaved, the self-destructive spacer will spontaneously dissociate from the second portion (e.g., the coagulation factor or procoagulant peptide).

[1275] In some respects, examples of self-destructive spacers are optionally substituted p-aminobenzyl alcohol (PAB) moiety, o-aminobenzyl acetal or p-aminobenzyl acetal, or other aromatic compounds that are electronically similar to the PAB group (i.e., PAB type) such as 2-aminoimidazol-5-methanol derivatives (see, for example, Hay et al., 1999, Bioorg. Med. Chem. Lett. [Bioorganic and Medicinal Chemistry Letters] 9:2237) or those in which the phenyl group of the p-aminobenzyl alcohol (PAB) moiety is replaced by a heteroarylene.

[1276] In some embodiments, Y is Wherein, the variables are as defined in the context; in other embodiments, Y is , where the variables are defined as in the context.

[1277] In some embodiments, Y is selected from: (AM); in other embodiments, Y is (PABC).

[1278] V. Drugs

[1279] The terms “drug fraction,” “drug payload,” “cytotoxic payload,” “therapeutic molecule,” “therapeutic payload,” “therapeutic agent,” and “therapeutic fraction,” which are used interchangeably in this article, refer to the chemical or biological fraction conjugated to an antibody.

[1280] In some respects, the free drug incorporated into the drug moiety is a cytotoxic compound, typically a cytotoxic compound with an aliphatic secondary amine as a coupling stalk.

[1281] The following provides examples of drugs that can be used with ADCs, namely, drugs that can be conjugated with antibodies, including antibiotics, DNA synthesis inhibitors, RNA polymerase II inhibitors and RNA splice inhibitors, mitotic inhibitors (e.g., tubulin inhibitors and microtubule polymerization inhibitors), antitumor antibiotics, immunomodulators, gene therapy vectors, alkylating agents, antiangiogenic agents, antimetabolites, boron-containing agents, chemoprotective agents, hormonal agents, glucocorticoids, photoactive therapeutic agents, oligonucleotides, radioisotopes, radiosensitizers, topoisomerase inhibitors (e.g., topoisomerase I inhibitors), tyrosine kinase inhibitors, immunomodulators, and chelating ligands and combinations thereof.

[1282] In some embodiments, microtubule inhibitors and microtubule polymerization inhibitors, antibiotics, DNA synthesis inhibitors, topoisomerase I inhibitors, RNA polymerase II inhibitors, and RNA spliceosome inhibitors are preferred.

[1283] In some embodiments, the pharmaceutical portion is a tubulin inhibitor and a microtubule polymerization inhibitor. In some particular embodiments, the pharmaceutical portion is oligristatin. In some particular embodiments, the pharmaceutical portion is a maytansine. In some particular embodiments, the pharmaceutical portion is tubulolysin. In some particular embodiments, the pharmaceutical portion is candidacin. In some particular embodiments, the pharmaceutical portion is eribulin. In some particular embodiments, the pharmaceutical portion is rhizobium.

[1284] In some embodiments, the pharmaceutical portion is an antibiotic. In some particular embodiments, the pharmaceutical portion is chachomycosis. In some particular embodiments, the pharmaceutical portion is doxorubicin. In some particular embodiments, the pharmaceutical portion is an anthracycline.

[1285] In some embodiments, the pharmaceutical portion is a DNA synthesis inhibitor. In some particular embodiments, the pharmaceutical portion is pyromalacin. In some particular embodiments, the pharmaceutical portion is PBD (pyrrolobenzodiazepine). In some particular embodiments, the pharmaceutical portion is IGN (indolinobenzodiazepine).

[1286] In some embodiments, the pharmaceutical portion is a topoisomerase I inhibitor. In some particular embodiments, the pharmaceutical portion is camptothecin. In some particular embodiments, the pharmaceutical portion is a camptothecin analogue.

[1287] In some embodiments, the pharmaceutical portion is an RNA polymerase II inhibitor. In some particular embodiments, the pharmaceutical portion is muscarine.

[1288] In some embodiments, the pharmaceutical portion is an RNA spliceosome inhibitor selected from the group consisting of sprisstatin and tylansstatin.

[1289] In some embodiments, cinnamic acid alkaloids (maytansin, maytanol, DM1, DM2, DM3, DM4, maytansin and anthraquinone) and their analogues are preferred.

[1290] In some embodiments, the pharmaceutical component is an immunomodulator.

[1291] In some embodiments, the immunomodulator is a TLR7 agonist. In some embodiments, the TLR7 agonist is imidazoquinoline, imidazoquinolineamine, thiazoquinoline, aminoquinoline, aminoquinazoline, pyrido[3,2-d]pyrimidine-2,4-diamine, pyrimidine-2,4-diamine, 2-aminoimidazole, 1-alkyl-1H-benzimidazole-2-amine, tetrahydropyridopyrimidine, heteroarylthiadiazine-2,2-dioxide, benzonaphthylidine, guanosine analog, adenosine analog, thymidine homopolymer, ssRNA, CpG-A, polyG10, or polyG3.

[1292] In some embodiments, the immunomodulator is a TLR8 agonist. In some embodiments, the TLR8 agonist is selected from imidazoquinoline, thiazoquinoline, aminoquinoline, aminoquinazoline, pyrido[3,2-d]pyrimidine-2,4-diamine, pyrimidine-2,4-diamine, 2-aminoimidazole, 1-alkyl-1H-benzimidazole-2-amine, tetrahydropyridopyrimidine, or ssRNA.

[1293] In some embodiments, the immunomodulator is a STING agonist.

[1294] In some embodiments, the immunomodulator is a RIG-I agonist.

[1295] In some embodiments, the RIG-I agonist is selected from KIN1148, SB-9200, KIN700, KIN600, KIN500, KIN100, KIN101, KIN400 and KIN2000.

[1296] In some embodiments, the drug portion is a chelating ligand.

[1297] In some embodiments, the chelating ligand is selected from platinum (Pt), ruthenium (Ru), rhodium (Rh), gold (Au), silver (Ag), copper (Cu), molybdenum (Mo), titanium (Ti), or iridium (Ir); radioactive isotopes, such as yttrium-88, yttrium-90, technetium-99, copper-67, rhenium-188, rhenium-186, gallium-66, gallium-67, indium-111, indium-114, indium-115, lutetium-177, strontium-89, samarium-153, and lead-212.

[1298] In a particular embodiment, the pharmaceutical portion is a DXD. The DXD is represented by the following structural formula, wherein... This indicates the attachment site to the connector.

[1299] DXD

[1300] In a particular embodiment, the pharmaceutical component is eczemab. Ecinotemab is represented by the following structural formula, wherein... This indicates the attachment site to the connector.

[1301] Iciticon

[1302] In a particular embodiment, the pharmaceutical portion is monomethylolpropamine E (also known as MMAE). MMAE is represented by the following structural formula, wherein... This indicates the attachment site to the connector.

[1303] MMAE

[1304] In a particular embodiment, the pharmaceutical component is monomethylolpropamine F (also known as MMAF). MMAF is represented by the following structural formula, wherein... This indicates the attachment site to the connector.

[1305] MMAF

[1306] In a particular embodiment, the pharmaceutical component is SG3199, which is a cytotoxic DNA minor groove interstrand crosslinked pyrrolobenzodiazepine (PBD) dimer. SG3199 is represented by the following structural formula, wherein... This indicates the attachment site to the connector.

[1307] SG3199

[1308] In a particular embodiment, the pharmaceutical part is 7-ethyl-10-hydroxycamptothecin (also known as SN38). SN38 is represented by the following structural formula, wherein... This indicates the attachment site to the connector.

[1309] or SN38

[1310] In a particular embodiment, the pharmaceutical part is N2'-deacetylated-N2'-(3-mercapto-1-oxopropyl)-matansine (also known as DM1). DM1 is represented by the following structural formula, wherein... This indicates the attachment site to the connector.

[1311] DM1

[1312] In some specific embodiments, the maytansin alkaloid pharmaceutical moiety is N2'-deacetylated-N2'-(4-methyl-4-mercapto-1-oxopentyl)-matansin (also known as DM4). DM4 is represented by the following structural formula, wherein... This indicates the attachment site to the connector.

[1313] DM4.

[1314] VI. Pharmaceutical Compositions

[1315] This disclosure also provides compositions, such as pharmaceutical compositions, containing conjugates of this disclosure formulated with pharmaceutically acceptable carriers.

[1316] The therapeutic formulations disclosed herein can be prepared by mixing a conjugate having the desired purity with an optional physiologically acceptable carrier, excipient, or stabilizer in the form of a lyophilized formulation or an aqueous solution. The acceptable carrier, excipient, or stabilizer is non-toxic to the receptor at the dose and concentration used and includes buffers (such as phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid and methionine); preservatives (e.g., octadecyl dimethyl benzyl ammonium chloride; hexamethyl ammonium chloride; benzalkonium chloride; benzyl chloride; phenol, butanol, or benzyl alcohol; alkyl esters of p-hydroxybenzoate, such as methylparaben or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); and low molecular weight (less than about 10 amino acids). Proteins containing amino acid residues, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates; chelating agents, such as EDTA; sugars, such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions, such as sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants, such as Tween, Pluronics, or PEG.

[1317] The formulation may also contain one or more active compounds necessary for the specific indication being treated, preferably those with complementary activities that will not adversely affect each other. For example, the formulation may further contain another antibody or bispecific antibody, cytotoxic agent, chemotherapeutic agent, or ADC. Such molecules are suitably present in a combination in amounts effective for the intended purpose.

[1318] Active ingredients can also be encapsulated in microcapsules, for example, prepared by coagulation techniques or by interfacial polymerization, such as hydroxymethyl cellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules) or in crude emulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences, 16th edition, Osol, A. (ed. 1980).

[1319] The pharmaceutical compositions disclosed herein can be used in combination therapy, i.e., in combination with other pharmaceutical agents. Examples of therapeutic agents that can be used in combination therapy are described in more detail below.

[1320] Preparations intended for in vivo administration must be sterile. This can be readily achieved through filtration using a sterile filter membrane. Sterile injectable solutions can be prepared by incorporating the desired amount of the active compound with a combination of one or more of the aforementioned components (as needed) into a suitable solvent, followed by sterile microfiltration. Typically, dispersions are prepared by incorporating the active compound into a sterile medium containing a base dispersion medium and other desired components from those listed above. In the case of sterile powders used to prepare sterile injectable solutions, preferred methods of preparation include vacuum drying and freeze-drying (lyophilization), which produce powders of the active ingredient and any other desired components from their previously sterile filtered solutions.

[1321] VII. Dosage

[1322] The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending on the subject being treated and the specific route of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form is typically the amount of the composition that produces the therapeutic effect. Generally, this amount ranges from about 0.01% to about 99% of the active ingredient, preferably from about 0.1% to about 70%, and most preferably from about 1% to about 30% of the active ingredient combined with a pharmaceutically acceptable carrier, within one hundred percent.

[1323] Dosing regimens are adjusted to provide the best expected response (e.g., a therapeutic response). For example, the medication may be administered as a single bolus, administered in multiple doses over time, or the dose may be proportionally reduced or increased depending on the urgency of the treatment situation. Formulating parenteral compositions in unit dosage form is particularly advantageous for ease of administration and uniform dosing. As used herein, unit dosage form refers to a physically discrete unit suitable as a single dose to a subject to be treated; each unit contains a predetermined amount of the active compound, calculated to produce the desired therapeutic effect associated with the desired drug delivery. The specifications of the unit dosage form disclosed herein are determined by and directly depend on: (a) the unique characteristics of the active compound and the specific therapeutic effect to be achieved, and (b) the inherent limitations in the area of ​​sensitivity of the individual to which such active compound is combined for treatment.

[1324] For the administration of the conjugates disclosed herein, the dosage range is from about 0.0001 to 100 mg / kg of host body weight, and more typically from 0.01 to 50 mg / kg of host body weight. For example, the dosage may be 0.3 mg / kg body weight, 1 mg / kg body weight, 3 mg / kg body weight, 5 mg / kg body weight, or 10 mg / kg body weight, or in the range of 1-10 mg / kg. Exemplary treatment regimens require daily administration, twice weekly, once weekly, once every two weeks, once every three weeks, once every four weeks, once monthly, once every three months, or once every three to six months. Preferred administration regimens of the conjugates disclosed herein include intravenous administration of 1 mg / kg body weight or 3 mg / kg body weight, wherein the conjugate is administered using one of the following administration regimens: (i) once every four weeks for a total of six doses, followed by once every three months; (ii) once every three weeks; (iii) once 3 mg / kg body weight, followed by 1 mg / kg body weight every three weeks.

[1325] Alternatively, the conjugate can be administered as a sustained-release formulation, in which case a lower frequency of administration is required. The dosage and frequency vary depending on the half-life of the conjugate in the patient. Typically, human antibodies exhibit the longest half-life, followed by humanized antibodies, chimeric antibodies, and non-human antibodies. The dosage and frequency of administration can vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic use, relatively low doses are administered at relatively infrequent intervals over a long period. Some patients continue treatment for the rest of their lives. In therapeutic use, relatively high doses are sometimes required at relatively short intervals until disease progression decreases or ceases, and preferably until the patient shows partial or complete improvement in disease symptoms. Thereafter, a prophylactic regimen can be administered to the patient.

[1326] The actual dose level of the active ingredient in the pharmaceutical compositions disclosed herein can be varied to obtain an amount of active ingredient that is effective in achieving the desired therapeutic response and is non-toxic to the patient for a particular patient, composition, and route of administration. The selected dose level will depend on a variety of pharmacokinetic factors, including the activity of the particular composition used herein, the route of administration, the time of administration, the excretion rate of the particular compound used, the duration of treatment, other drugs, compounds, and / or materials used in combination with the particular composition used, and factors well known in the medical field such as the age, sex, weight, disease condition, general health status, and medical history of the patient being treated.

[1327] The "therapeutic effective dose" of the ADC disclosed herein preferably results in a reduction in the severity of disease symptoms, an increase in the frequency and duration of symptom-free periods, or prevention of injury or disability due to disease suffering. For example, in the treatment of tumors, the "therapeutic effective dose" preferably inhibits cell growth or tumor growth or metastasis by at least about 20%, more preferably at least about 40%, even more preferably at least about 60%, and still more preferably at least about 80%, relative to an untreated subject. The ability of the agent or compound to inhibit tumor growth can be evaluated in animal model systems that predict efficacy against human tumors. Alternatively, this property of the composition can be evaluated by examining the ability of the compound to inhibit (such in vitro inhibition) using assays known to those skilled in the art. A therapeutically effective amount of a therapeutic compound can reduce tumor size, metastasis, or otherwise improve symptoms in a subject. Those skilled in the art will be able to determine such amounts based on factors such as the subject's body size, the severity of the subject's symptoms, and the specific composition or route of administration chosen.

[1328] VIII. Application

[1329] The compositions disclosed herein can be administered via one or more routes of administration using one or more of a variety of methods known in the art. As will be understood by those skilled in the art, the route and / or manner of administration will vary depending on the desired results. Preferred routes of administration for the conjugates disclosed herein include intravenous, intramuscular, intradermal, intraperitoneal, subcutaneous, spinal, or other parenteral administration routes, such as by injection or infusion. As used herein, the phrase “parenteral administration” refers to administration modes that are typically administered by injection, other than enteral and local administration, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrasheath, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, tracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions. Alternatively, the conjugates disclosed herein can be administered via non-parenteral routes, such as local, epidermal, or mucosal administration routes, such as intranasal, oral, vaginal, rectal, sublingual, or local administration.

[1330] These active compounds can be prepared using carriers that protect the compound from rapid release, such as controlled-release formulations, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. Many methods for preparing such formulations are patented or well known to those skilled in the art. See, for example, Sustained and Controlled Release Drug Delivery Systems, JR Robinson, editor, Marcel Dekker, Inc., New York, 1978.

[1331] The therapeutic compositions can be administered using medical devices known in the art. For example, the therapeutic compositions disclosed herein can be administered using needle-free subcutaneous injection devices, such as those disclosed in US 5399163, US 5383851, US 5312335, US5064413, US 4941880, US 4790824, and US 4596556. Examples of well-known implants and modules used in this disclosure include those described in US 4487603, US 4486194, US 4447233, US 4447224, US4439196, and US 4475196. These patents are incorporated herein by reference. Many other such implants, delivery systems, and modules are known to those skilled in the art.

[1332] IX. Treatment Methods

[1333] In one aspect, this disclosure relates to the use of the aforementioned conjugate in vivo to treat a disease in a subject. In one embodiment, this disclosure provides a method for preventing and / or treating a disease in a subject in need, the method comprising administering to the subject a therapeutically effective amount of the conjugate or a composition comprising the conjugate.

[1334] In some embodiments, the disease is selected from cancer, infectious diseases, inflammatory diseases, autoimmune diseases, and immunodeficiency diseases.

[1335] In one particular embodiment, the disease is cancer.

[1336] Non-limiting examples of preferred cancers for treatment include head and neck cancer, esophageal cancer, lung cancer, colon cancer, rectal cancer, stomach cancer, pancreatic cancer, ovarian cancer, prostate cancer, breast cancer, leukemia, myeloma, squamous cell carcinoma, melanoma, brain cancer, cervical cancer, liver cancer, bladder cancer, breast cancer, kidney cancer, testicular cancer, and thyroid cancer.

[1337] Non-limiting examples of preferred autoimmune diseases to be treated include systemic sclerosis and atherosclerosis.

[1338] On the other hand, this disclosure relates to treating a subject in vivo with the aforementioned conjugate to inhibit the growth and / or metastasis of cancerous tumors. In one embodiment, this disclosure provides a method for inhibiting the growth of tumor cells in a subject and / or limiting the metastatic spread of tumor cells, the method comprising administering a therapeutically effective amount of the conjugate to the subject.

[1339] As used herein, the term "subject" is intended to include both humans and non-human animals. Non-human animals include all vertebrates, such as mammals and non-mammals, such as non-human primates, sheep, dogs, cats, cattle, horses, chickens, amphibians, and reptiles, although mammals are preferred, such as non-human primates, sheep, dogs, cats, cattle, and horses. Preferred subjects include human patients who require enhanced immune responses. These methods are particularly suitable for treating human patients with disorders that can be treated by enhancing immune responses.

[1340] The above treatments can also be combined with standard cancer treatments. For example, it can be effectively combined with chemotherapy regimens. In these cases, the dosage of the chemotherapeutic agents can be reduced (Mokyr, M. et al. Cancer Res., 1998, 58, 5301–5304).

[1341] Other antibodies that can activate host immune responses can be used in or in conjunction with the bispecific molecular drug conjugates disclosed herein. These include molecules that target the surface of dendritic cells, activating DC function and antigen presentation. For example, anti-CD40 antibodies can effectively displace T cell helper activity (Ridge, J. et al., Nature, 1998, 393, 474–478) and can be used in combination with the bispecific molecular drug conjugates disclosed herein (Ito, N. et al., Immunobiology, 2000, 201, 527–540). Similarly, antibodies targeting T cell co-stimulatory molecules, such as CTLA-4 (US 5811097), CD28 (Haan, J. et al. Immunol. Lett. [Immunology Letters], 2014, 162, 103–112), OX-40 (Weinberg, A. et al. J. Immunol. [Journal of Immunology], 2000, 164, 2160–2169), 4-1BB (Melero, I. et al. Nature Med. [Nature Medicine], 1997, 3, 682–685) and ICOS (Hutloff, A. et al. Nature [Nature], 1999, 397, 262–266) or antibodies targeting PD-1 (US 8008449) and antibodies targeting PD-L1 (US 8008449) are also effective. 7943743; US8168179) can also provide increased T-cell activation levels. In another instance, the bispecific molecular drug conjugates disclosed herein can be used in combination with antitumor antibodies such as Rituxan (rituximab), Herceptin (trastuzumab), BEXXAR (tosimomab), ZEVALIN (teimomab), CAMPATH (alemumab), LYMPHOCIDE (epizumab), AVASTIN (bevacizumab), and TARCEVA (erlotinib).

[1342] Synthesis method

[1343] The compounds and methods of the present invention will be better understood in conjunction with the following description.

[1344] Example

[1345] The compounds and methods of the present invention will be better understood in conjunction with the following examples, which are for illustrative purposes only and not for limiting the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art, and such changes and modifications (including, but not limited to, those involving the chemical structures, substituents, derivatives, formulations, and / or methods of the present invention) can be made without departing from the spirit of the invention and the scope of the appended claims.

[1346] General methods

[1347] Will 1 ¹H NMR and other NMR spectra were recorded on a Bruker Ascend 400 spectrometer. Chemical shifts are expressed in parts per million (ppm). Coupling constants are in Hertz (Hz). Splitting modes describe the apparent multiplicity and are designated as s (singleton), d (doublet), t (triplet), q (quartet), quint (quintet), m (multiplet), and br (broad peak).

[1348] Analytical low-resolution mass spectrometry (MS) was performed on an Agilent 1290 equipped with an SQ detector using a Waters Xbridge C18, 4.6 x 50 mm, 3.5 µm, and recorded using gradient elution.

[1349] Mobile phase A: Solvent A: 0.1% TFA in water; Solvent B: 0.1% TFA in acetonitrile; 5%-95% B for 1.3 min.

[1350] Mobile phase B: Solvent A: 0.1% FA in water; Solvent B: 0.1% FA in acetonitrile; 5%-95% B for 2.0 min.

[1351] Run preparative high-performance liquid chromatography (Prep-HPLC) on Gilson or Waters under the following conditions:

[1352] Method A: Waters SunFire 10 pm C18 column (100 A, 250 x 19 mm). Solvent A was water / 0.01% trifluoroacetic acid (TFA) and solvent B was acetonitrile. Elution conditions were as follows: solvent B was increased linearly from 5% to 100% at a flow rate of 30 mL / min over 20 minutes.

[1353] Method B: Waters SunFire 10 pm C18 column (100 A, 250 x 19 mm). Solvent A was water / 0.05% formic acid (FA) and solvent B was acetonitrile. Elution conditions were as follows: solvent B was increased linearly from 5% to 100% at a flow rate of 30 mL / min over 20 minutes.

[1354] Method C: Waters Xbridge 10 pm C18 column (100 A, 250 x 19 mm). Solvent A was water / 10 mM ammonium bicarbonate (NH4HCO3) and solvent B was acetonitrile. Elution conditions were as follows: solvent B was increased linearly from 5% to 100% at a flow rate of 30 mL / min over 20 minutes.

[1355] The compounds and methods of the present invention will be better understood in conjunction with the following examples, which are for illustrative purposes only and not for limiting the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art, and such changes and modifications (including, but not limited to, those involving the chemical structures, substituents, derivatives, formulations, and / or methods of the present invention) can be made without departing from the spirit of the invention and the scope of the appended claims.

[1356] Example 1: Preparation of LP1 (MC-VGR-DXD)

[1357]

[1358] 6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-N-((7S,13S)-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl)amino)-7-(3-guanidinopropyl)-14-methyl-1,6,9,12-tetraoxo-3-oxa-5,8,11-triazapentadecan-13-yl)hexamamide

[1359]

[1360] Step 1a. N,N,N',N'-tetramethylchloromethacin hexafluorophosphate (8.4 g, 30.0 mmol) and 1-methylimidazole (4.1 g, 50.0 mmol) were added to a mixture of (((9H-fluorene-9-yl)methoxy)carbonyl)-L-arginine (CAS: 91000-69-0) (8.0 g, 20.0 mmol) and glycine tert-butyl hydrochloride (2.62 g, 20.0 mmol) in DMF (120 mL). The mixture was stirred at 30 °C for 12 h and concentrated under reduced pressure. The residue was purified to provide the desired compound (4.32 g, 41% yield) as a white solid. LC-MS: (ESI) m / z (M+H), 510.3.

[1361] Step 1b. A mixture of the compound from Step 1a (8.0 g) and TFA (30 mL) in DCM (80 mL). The mixture was stirred at 30 °C for 3 h. The filtrate was concentrated under reduced pressure to give a crude product (8.0 g) as a gray solid. LC-MS: (ESI) m / z (M+H), 454.2.

[1362] Step 1c. The mixture of the compound from Step 1b (4.4 g, 9.73 mmol) and Cu(OAc)₂ (1.14 g, 0.973 mmol) and Pb(OAc)₄ (6.5 g, 14.6 mmol) in THF (40 mL) was stirred at 30 °C for 4 h. The organic solvent was removed under reduced pressure. The residue was then subjected to EtOAc (3 Divide between 100 mL of water and 300 mL of water. The residue was washed with 10 mL of aqueous NH4Cl (saturated 200 mL). The combined organic matter was dried over anhydrous Na2SO4. The filtrate was concentrated under vacuum to give the desired compound (5.0 g) as a white solid. LC-MS: (ESI) m / z (M+H), 468.2.

[1363] Step 1d. The mixture of the compound from Step 1c (6.82 g, 14.6 mmol), benzyl 2-hydroxyacetate (12.2 g, 73 mmol), and pyridinium p-toluenesulfonate (3.7 g, 14.6 mmol) in THF (120 mL) was stirred at 25 °C for 24 h. The organic solvent was removed under reduced pressure. The residue was purified to provide the desired compound (800 mg, 10% yield) as a white solid. LC-MS: (ESI) m / z (M+H), 574.2.

[1364] Step 1e. The compound (800 mg) from Step 1d was stirred at 30°C for 5 h in a mixture of EtOAc (10 mL), methanol (10 mL), and 10% Pd / C (0.5 g). The filtrate was rinsed with methanol (20 mL). 3) Wash and concentrate under reduced pressure to provide a crude product (1.0 g) as a white solid. LC-MS: (ESI) m / z (M+H), 484.2.

[1365] Step 1f. A mixture of the compound from Step 1e (966 mg, 2.04 mmol), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholino-4-onium chloride (452 ​​mg, 1.632 mmol), eczetidine mesylate (CAS169869-90-3) (870 mg, 1.632 mmol), and DIPEA (792 mg, 6.12 mmol) in DMF (8 mL) was stirred at 30 °C for 1 h. The mixture was quenched with water (0.5 mL), concentrated, and the residue was purified by preparative HPLC (Xbridge-C18, CH3CN / H2O (0.1% NH3)) to give the desired compound (990 mg, 55% yield) as a white solid. LC-MS: (ESI) m / z (M+H), 910.4.

[1366] Step 1g. The mixture of the compound from Step 1f (628 mg, 0.698 mmol) and diethylamine (2 mL) in DCM (10 mL) was stirred at 30 °C for 2 h. It was quenched with water (0.5 mL), concentrated, and the residue was purified by preparative HPLC (Xbridge-C18, CH3CN / H2O (0.1% NH3)) to give the desired compound (350 mg, 74% yield) as a pale yellow oil. LC-MS: (ESI) m / z (M+H), 679.4.

[1367] Step 1 h. A mixture of 2-[(2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-3-methylbutyramide]acetic acid (CAS 142810-19-3) (1.0 g, 2.52 mmol) in DMF (80 mL) and DBU (403 mg) was stirred at 30 °C for 3 h. It was concentrated to give a crude product, which could proceed to the next step without further purification. A mixture of the crude product, 2,5-dioxopyrrolidone-1-yl 6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)hexanoate (932 mg) and DIPEA (423 mg) in DMF (4 mL) was stirred at 30 °C for 2 h. The residue was concentrated and purified by preparative HPLC (Xbridge-C18, CH3CN / H2O (0.1% NH3)) to give the desired compound as a white solid (466 mg, 50% yield). LC-MS: (ESI) m / z (M+H), 368.2.

[1368] Step 1i. A mixture of the compound from Step 1g (25 mg, 0.036 mmol), the compound from Step 1h (15 mg, 0.041 mmol), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholino-4-onium chloride (10 mg, 0.036 mmol), and triethylamine (19 mg, 0.18 mmol) in DMF (1 mL) was stirred at 30 °C for 18 h. The filtrate was concentrated under reduced pressure and purified by preparative HPLC (C18, H2O (containing 0.1% formic acid) in 30%–53% acetonitrile) to provide the title compound (12 mg, 32% yield) as a pale yellow solid. LC-MS: (ESI) m / z (M+H), 1028.4. 1HNMR (400 MHz, DMSO+D2O) δ 7.79 (d, J = 8.0 Hz, 1H), 7.34 (s, 1H), 6.97 (s,2H), 5.60-5.58 (m, 1H), 5.41 (q, J = 7.6 Hz, 2H), 5.20 (q, J = 7.2 Hz, 2H),4.58-4.65 (m, 2H), 4.22 (t, J = 8.0 Hz, 1H), 4.01-4.03 (m, 3H), 3.72 (s, 2H),3.36 (t, J = 4.0 Hz, 2H), 3.08-3.16 (m, 2H), 3.07 (t, J = 4.0 Hz, 2H), 2.40(s, 3H), 2.02-2.19 (m, 3H), 1.87 – 1.66 (m, 3H), 1.52-1.45 (m, 7H), 1.24 –1.17 (m, 4H), 0.89-0.82 (m, 9H).

[1369] Example 2: Preparation of LP2 (MC-SGR-DXD)

[1370]

[1371] 6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-N-((7S,13S)-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl)amino)-7-(3-guanidinopropyl)-14-hydroxy-1,6,9,12-tetraoxo-3-oxa-5,8,11-triazatetradecane-13-yl)hexanoamide

[1372]

[1373] Step 2a. A mixture of benzyloxycarbonyl-O-benzyl-L-serine (CAS 20806-43-3) (6.0 g, 18.2 mmol), glycine tert-butyl ester (2.4 g, 18.2 mmol), HATU (10.4 g, 27.3 mmol), and DIPEA (6.36 mL, 36.4 mmol) in DMF (80 mL) was stirred at 30 °C for 1 h. The mixture was quenched with water (150 mL), concentrated, and TFA (20 mL) was added to the residue in DCM (80 mL). The mixture was stirred at 30 °C for 12 h. It was concentrated to give the desired compound (6.5 g, 92% yield) as a yellow solid. LC-MS: (ESI) m / z (M+H), 387.2.

[1374] Step 2b. Under a hydrogen atmosphere, the mixture of the compound from step 2a (3.8 g, 9.8 mmol) in methanol (35 mL) and 10% Pd / C (0.38 g) was stirred at 30 °C for 18 h. The filtrate was then treated with MeCN / water (1:1, 10 mL). 3) Wash and concentrate under reduced pressure to provide the desired compound as a white solid (1.1 g, 68% yield).

[1375] Step 2c. The compound from step 2b (1.1 g, 6.8 mmol), 2,5-dioxopyrrolidone-1-yl 6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)hexanoate (3.13 g, 10.2 mmol), and DIPEA (2.36 g, 13.6 mmol) were stirred at 30 °C for 6 h in a mixture of DMF (3 mL), MeCN (3 mL), and water (3 mL). The mixture was concentrated, and the residue was purified by preparative HPLC (Xbridge-C18, CH3CN / H2O (0.1% NH3)) to give the desired compound (1.0 g, 42% yield) as a white solid. LC-MS: (ESI) m / z (M+H), 356.2.

[1376] Step 2d. A mixture of the compound from step 1g (25 mg, 0.0393 mmol), the compound from step 2c (18 mg, 0.0472 mmol), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholine-4-onium chloride (18 mg, 0.0472 mmol), and DIPEA (16 mg, 0.12 mmol) in DMF (1 mL) was stirred at 30 °C for 4 h. The filtrate was concentrated under reduced pressure and purified by preparative HPLC (C18, H2O (containing 0.1% formic acid) in 30%–53% acetonitrile) to provide the title compound (11 mg, 28% yield) as a yellow solid. LC-MS: (ESI) m / z (M+H), 1016.4. 1 H NMR(400 MHz, DMSO+D2O) δ 8.76 (t, J = 8.0 Hz, 1H), 8.55 (d, J = 8.0 Hz, 1H), 7.95 (d, J = 8.0 Hz, 1H), 7.77 (d, J = 12.0 Hz, 1H), 7.38 – 7.30 (m, 1H),7.34 (s, 1H), 6.97 (s, 2H), 5.58 (q, J = 8.0 Hz, 1H), 5.41 (d, J = 4.0 Hz,2H), 5.19 (d, J = 4.0 Hz, 2H), 4.63 (d, J = 4.0 Hz, 2H) 4.19 – 4.21 (m, 2H),4.01 (s, 2H), 3.45 – 3.39 (m, 2H), 3.25 – 3.11 (m, 2H), 3.02-3.24 (m, 3H),2.42 (s, 3H), 2.11 – 2.18 (m, 3H), 1.85-1.88 (m, 2H), 1.53 – 1.46 (m, 7H), 1.24 – 1.18 (m, 4H), 0.87 (t, J = 7.6 Hz, 3H).

[1377] Example 3: Preparation of LP3 (MC-SGC-DXD)

[1378]

[1379] 6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-N-((7S,13S)-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl)amino)-14-hydroxy-1,6,9,12-tetraoxo-7-(3-ureopropyl)-3-oxa-5,8,11-triazatetradecane-13-yl)hexamamide

[1380]

[1381]

[1382] Step 3a. At 0 °C, add [chloro(dimethylamino)methylene]dimethylammonium hexafluoro-λ5-phosphine salt (7.77 g, 27.678 mmol) and 1-methylimidazole (4.212 mL, 52.839 mmol) to a solution of (2S)-5-[(aminocarbonyl)amino]-2-({[(9H-fluorene-9-ylmethyl)oxy]carbonyl}amino)valerate (10.0 g, 25.2 mmol) in DMF (100 mL). Stir the mixture for 10 minutes. Add 2-methylpropyl-2-ylaminoacetic acid ester (3.47 g, 26.420 mmol) to the mixture and stir the mixture at 15 °C for 12 hours. Concentrate the mixture under reduced pressure and partition the residue between DCM (100 mL) and water (50 mL). Dilute the aqueous layer with DCM (50 mL) 2) Extraction was performed, and the combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid chromatography (160 g silica column, DCM / MeOH, where MeOH ranged from 0–8%) to provide the desired product (8.8 g, white solid, yield: 69%). LC-MS: (ESI) m / z (M+H), 511.2.

[1383] Step 3b. At 0 °C, 2,2,2-trifluoroacetic acid (19.8 mL, 258.5 mmol) in DCM (80 mL) was added to a solution of the compound from Step 3a (8.8 g, 17.2 mmol) for 5 minutes. The mixture was allowed to warm naturally to 20 °C and then stirred for 12 hours. The mixture was concentrated under reduced pressure and the residue was dissolved in DMF (20 mL). The solution was purified by reversed-phase column chromatography (220 g C18 gel column, H2O / CH3CN, where CH3CN ranged from 0–16%) to provide the desired product (4.5 g, white solid, yield: 58%). LC-MS: (ESI) m / z (M+H), 455.2.

[1384] Step 3c. Add saline tetraacetate (1.6 g, 3.6 mmol) to a mixture of the compound from step 3b (1.1 g, 2.4 mmol), copper(II) oxoalkyl diacetate (140 mg, 0.73 mmol), and acetic acid (1.0 mL, 17.0 mmol) in THF (40 mL). Stir the mixture at 30 °C under N2 for 2 hours. Filter the solution and purify the filtrate by passing it through a reversed-phase column (120 g C18 gel column, H2O / CH3CN, where CH3CN ranges from 0–35%) to provide the desired product (1.0 g, white solid, yield: 79%). LC-MS: (ESI) m / z (M+Na), 491.2.

[1385] Step 3d. At 0 °C, benzyl glycolate (709.4 mg, 4.3 mmol) in DCM (4 mL) was added to a mixture of the compound from Step 3c (400 mg, 0.85 mmol) and 2,2,2-trifluoroacetic acid (3.3 mL, 42.7 mmol) in DCM (4 mL) over 10 minutes. The mixture was stirred at 20 °C for 4 hours. The solution was filtered and the filtrate was purified by passing it through a reversed-phase column (220 g C18 gel column, H2O / CH3CN, where CH3CN ranged from 0–47%) to provide the desired product (210 mg, white solid, yield: 42.8%). LC-MS: (ESI) m / z (M+Na), 597.2.

[1386] Step 3e. Palladium (0) (111.12 mg, 0.104 mmol) was added to a mixture of the compound from step 3d (200 mg, 0.348 mmol) in MeOH (10 mL) and EA (10 mL). The mixture was stirred at 15 °C for 2 hours. The mixture was concentrated to obtain the title compound, which was ready for use in the next step without purification (0.16 g, white solid, quantitative yield). LC-MS: (ESI) m / z (M+Na), 507.2.

[1387] Step 3f. The mixture of the compound from Step 3e (150.40 mg, 0.310 mmol), (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,10,12,13,15-octahydrocyclohexane[1,2,3-de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-10,13-dione methanesulfonic acid (150 mg, 0.282 mmol) and DIPEA (0.14 mL, 0.847 mmol) in DMF (6 mL) was stirred at 0 °C for 2 min. 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-1,4-oxazinane-4-onium chloride hydrate (91.43 mg, 0.310 mmol) was added to the mixture. The mixture was stirred at 0 °C for 0.5 h. The solution was filtered and the filtrate was purified by passing it through a reversed-phase column (80 g C18 gel column, H2O / CH3CN, where CH3CN ranged from 0–46%) to provide the desired product (180 mg, white solid, yield: 70.7%). LC-MS: (ESI) m / z (M+H), 902.2.

[1388] Step 3g. Diethylamine (0.097 mL, 0.942 mmol) in DMF (1 mL) was added to a solution of the compound from Step 3f (85.0 mg, 0.094 mmol) at 0 °C for 2 h. The mixture was then allowed to warm naturally to 15 °C and stirred for 60 min. The mixture was concentrated to obtain the desired compound, which was ready for use in the next step without purification (64 mg, yellow solid, quantitative yield). LC-MS: (ESI) m / z (M+H), 680.4.

[1389] Step 3h. A mixture of N-[(2S)-2-{[6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-1-oxohexyl]amino}-3-hydroxypropionyl]glycine (60.0 mg, 0.17 mmol) and 1-hydroxytetrahydropyrrolo-2,5-dione (29.2 mg, 0.25 mmol) in DMF (1 mL) was stirred at 0 °C for 5 min. DCC (55.8 mg, 0.27 mmol) was added to the mixture. The mixture was naturally warmed to 15 °C and stirred for 12 h. The mixture was concentrated to obtain the desired compound, which was ready for use in the next step without purification (76.4 mg, white solid, quantitative yield). LC-MS: (ESI) m / z (M+H), 453.2.

[1390] Step 3i. Add the compound from step 3h (64.90 mg, 0.143 mmol) to a mixture of the compound from step 3g (65 mg, 0.096 mmol) and DIPEA (0.032 mL, 0.192 mmol) in DMF (2 mL). Stir the mixture at 15 °C for 30 min. Remove the solvent under vacuum and purify the residue by preparative HPLC (C18, H2O (containing 0.1% formic acid) in 30%–53% acetonitrile) to provide the title compound as a white solid (18.2 mg, 18.7% yield). LC-MS: (ESI) m / z (M+H), 1017.4. 1H NMR (400 MHz, DMSO) δ 8.65 (t, J = 6.4Hz, 1H), 8.44 (d, J = 8.8 Hz, 1H), 8.07 (t, J = 5.6 Hz, 1H), 7.84 (t, J = 6.8Hz, 2H), 7.72 (d, J = 10.8 Hz, 1H), 7.25 (s, 1H), 6.94-6.92 (m, 2H), 6.45 (s,1H), 5.83 (d, J = 6.4 Hz, 1H), 5.53-5.51 (m, 1H), 5.36 (s, 2H), 5.30 (s, 2H),5.14 (s, 2H), 4.92-4.89 (m, 1H), 4.57-4.51 (m, 2H), 4.15-4.11 (m, 2H), 3.92(s, 2H), 3.67-3.65 (m, 2H), 3.49-3.45 (m, 2H), 3.07-3.05 (m, 2H), 2.84-2.82(m, 2H), 2.33 (s, 3H), 2.08-2.03 (m, 3H), 1.86-1.74 (m, 3H), 1.57-1.55 (m,2H), 1.42-1.38 (m, 4H), 1.26-1.24 (m, 2H), 1.18-1.03 (m, 4H), 0.80 (t, J =7.2 Hz, 3H).

[1391] Example 4: Preparation of LP4 (MC-SAC-DXD)

[1392]

[1393] 6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-N-((7S,10S,13S)-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl)amino)-14-hydroxy-10-methyl-1,6,9,12-tetraoxo-7-(3-ureopropyl)-3-oxa-5,8,11-triazatetradecane-13-yl)hexamamide

[1394]

[1395] Step 4a. At 0 °C, HATU (2.97 g, 7.824 mmol), DIPEA (2.586 mL, 15.648 mmol), and 2-methylpropyl-2-yl alanine ester (1.25 g, 8.606 mmol) were slowly added to a solution of (2S)-2-({[(9H-fluorene-9-ylmethyl)oxy]carbonyl}amino)-3-[(2-methylprop-2-yl)oxy]propionic acid (3 g, 7.824 mmol) in DMF (20 mL). The resulting mixture was stirred at 15 °C for about 16 h. It was concentrated under reduced pressure and the residue was partitioned between EA (100 mL) and water (50 mL). The aqueous layer was extracted with EA (50 mL × 2), and the combined organic matter was dried (Na2SO4), filtered, and concentrated. The residue was purified by chromatography (silica, 0-10% DCM in MeOH) to provide the desired compound (3.5 g, white solid, 87% yield). LC-MS: (ESI) m / z (M+Na), 533.2.

[1396] Step 4b. Diethylamine (2.006 mL, 19.388 mmol) was slowly added to a solution of the compound from step 4a (3.3 g, 6.463 mmol) in DMF (20 mL) at 0 °C. The resulting mixture was stirred at room temperature for about 1 h. It was concentrated under reduced pressure, and the mixture was concentrated to obtain the desired compound, which could be used in the next step without purification (1.86 g, pale yellow oil, quantitative yield). LC-MS: (ESI) m / z (M+H), 289.4.

[1397] Step 4c. At 0 °C, DIPEA (2.063 mL, 12.483 mmol) and 1-{1-[(2,5-dioxotetrahydro-1H-pyrrolo-1-yl)oxy]-1-oxohexyl-6-yl}pyrrolo-2,5-dione (2.31 g, 7.490 mmol) were slowly added to a solution of the compound from Step 4b (1.8 g, 6.242 mmol) in DCM (20 mL). The resulting mixture was stirred at room temperature for about 16 h. It was concentrated under reduced pressure and the residue was partitioned between EA (200 mL) and water (100 mL). The aqueous layer was extracted with EA (100 mL × 2), and the combined organic matter was dried (Na2SO4), filtered, and concentrated. The residue was purified by chromatography (0-10% DCM in silica, MeOH) to provide the desired compound (2.2 g, white solid, 73% yield). LC-MS: (ESI) m / z (M+Na), 504.2.

[1398] Step 4d. At 0 °C, TFA (3 mL, 10.694 mmol) was slowly added to a solution of the compound from step 4c (500 mg, 1.038 mmol) in DCM (10 mL). The resulting mixture was stirred at 15 °C for approximately 16 h. It was concentrated under reduced pressure and the residue was purified by RP-FC (C18, 5%–95% MeCN in H2O) to provide the desired compound (340 mg, white solid, 88% yield). LC-MS: (ESI) m / z (M+1), 370.2.

[1399] Step 4e. At 0 °C, 1-hydroxytetrahydropyrrole-2,5-dione (97.16 mg, 0.844 mmol) and DCC (174.19 mg, 0.844 mmol) were slowly added to a solution of the compound from Step 4d (150 mg, 0.422 mmol) in DMF (0.5 mL). The resulting mixture was stirred at 15 °C for approximately 2 h. The mixture was concentrated to obtain the desired compound, which was ready for use in the next step without purification (190 mg, white solid, quantitative yield). LC-MS: (ESI) m / z(M+Na), 467.2.

[1400] Step 4f. At 0 °C, DIPEA (0.033 mL, 0.199 mmol) and the compound from step 6e (61.76 mg, 0.132 mmol) were slowly added to a solution of the compound from step 3g (45 mg, 0.066 mmol) in DMF (1 mL). The resulting mixture was stirred at 15 °C for about 2 h. It was concentrated under reduced pressure and the residue was purified by preparative HPLC (C18, 20%–60% acetonitrile in H2O (containing 0.1% formic acid)) to provide the title compound (10.6 mg, yellow solid, 15.5% yield). LC-MS: (ESI) m / z (M+H), 1031.3. 1H NMR (400 MHz, DMSO) δ 8.55-8.52(m, 1H), 8.44-8.42 (m, 1H), 8.06-8.04 (m, 1H), 7.8-7.79 (m, 2H), 7.72-7.70(m, 1H), 7.24 (s, 1H), 6.92 (s, 2H), 6.58-6.24(m, 1H) 5.87 (s, 1H), 5.55-5.50(m, 1H), 5.36 (s, 2H), 5.13 (s, 2H), 4.59-4.50 (m, 2H), 4.24-4.22 (m, 1H),4.14-4.11 (m, 1H), 4.06-4.03 (m, 1H), 3.92 (s, 2H), 3.31-3.2 1H NMR (400 MHz, DMSO) δ 8.55-8.52 (m, 1H), 8.44-8.42 (m, 1H), 8.06-8.04 (m, 1H), 7.8-7.79 (m,2H), 7.72-7.70 (m, 1H), 7.24 (s, 1H), 6.92 (s, 2H), 6.58-6.24(m, 1H) 5.87 (s,1H), 5.55-5.50 (m, 1H), 5.36 (s, 2H), 5.13 (s, 2H), 4.59-4.50 (m, 2H), 4.24-4.22 (m, 1H), 4.14-4.11 (m, 1H), 4.06-4.03 (m, 1H), 3.92 (s, 2H), 3.31-3.27(m, 3H), 3.19-3.03 (m, 3H), 2.84-2.83 (m, 2H), 2.70 (s, 1H), 2.33 (s, 3H),2.14-2.08 (m, 2H), 2.05-2.02 (m, 2H), 1.81-1.77 (m, 2H), 1.55-1.52 (m, 1H),1.41-1.37 (m, 6H), 1.22-1.08 (m, H), 0.80 (t, J = 7.2 Hz, 3H).7 (m, 3H),3.19-3.03 (m, 3H), 2.84-2.83 (m, 2H), 2.70 (s, 1H), 2.33 (s, 3H), 2.14-2.08(m, 2H), 2.05-2.02 (m, 2H), 1.81-1.77 (m, 2H), 1.55-1.52 (m, 1H), 1.41-1.37(m, 6H), 1.22-1.08 (m, H), 0.80 (t, J = 7.2 Hz, 3H).

[1401] Example 5: LP5 (MC-S) t Preparation of Bu)GC-DXD

[1402]

[1403] 6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-N-((7S,13S)-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl)amino)-16,16-dimethyl-1,6,9,12-tetraoxo-7-(3-ureopropyl)-3,15-dioxa-5,8,11-triazaheptadecane-13-yl)hexamamide

[1404]

[1405] Step 5a. At 0 °C, DCC (75.22 mg, 0.36 mmol) and 1-hydroxytetrahydropyrrole-2,5-dione (41.96 mg, 0.36 mmol) in DMF (3 mL) were added to a solution of (S)-2-(3-(tert-butoxy)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)hexamido)propamido)acetic acid (100 mg, 0.24 mmol) for 6 hours. The solution was filtered and concentrated to obtain the desired compound (100 mg, white oil, quantitative yield), which could be used in the next step without purification. LC-MS: (ESI) m / z (M+Na), 531.2.

[1406] Step 5b. Add the compound from step 5a (45 mg, 0.08 mmol) to a mixture of the compound from step 3g (30 mg, 0.04 mmol), DIPEA (17.1 mg, 0.13 mmol), and DMF (2 mL). Stir the mixture at 0 °C for 2 hours. Remove the solvent under vacuum and purify the residue by preparative HPLC (C18, 20%–60% acetonitrile in H2O (containing 0.1% TFA)) to provide the title compound as a yellow solid (17.2 mg, 36.3% yield). LC-MS: (ESI) m / z (M+H), 1074.6. 1H NMR (400 MHz, DMSO) δ 8.77–8.70 (m, 1H), 8.55–8.51(m, 1H), 8.10–8.07 (m, 1H), 8.00–7.85 (m, 2H), 7.79–7.75 (m, 1H), 7.31 (s,1H), 6.99 (s, 2H), 5.95–5.92 (m, 1H), 5.62–5.58 (m, 1H), 5.43 (s, 2H), 5.21(s, 2H), 4.65–4.61 (m, 2H), 4.31 – 4.14 (m, 2H), 3.99 (s, 2H), 3.81 – 3.66(m, 2H), 3.29–3.18 (m, 8H), 2.91 (s, 3H), 2.45–2.40 (m, 3H), 2.20–2.12 (m,4H), 1.92 – 1.81 (m, 2H), 1.56 – 1.39 (m, 6H), 1.39–1.32 (m, 2H), 1.21 – 1.15 (m, 2H), 1.09 (s, 9H), 0.87 (t, J = 7.2 Hz, 3H).

[1407] Example 6: Preparation of LP6 (MC-SGO(Boc)-DXD)

[1408]

[1409] ((S)-4-(2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)hexamido)-3-hydroxypropamido)acetamido)-5-(((2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl)amino)-2-oxoethoxy)methyl)amino)-5-oxopentyl)tert-butyl carbamate

[1410]

[1411] Step 6a. At 20°C, DIPEA (4.27 g, 33.0 mmol) was added in a single batch to a solution of (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-5-((tert-butoxycarbonyl)amino)valerate (5.0 g, 11.0 mmol), benzyl 2-aminoacetate (1.82 g, 11.0 mmol), EDCI (2.53 g, 13.2 mmol), and HOBT (1.78 g, 13.2 mmol) in DCM (150 mL). The mixture was stirred at 20°C for 12 hours. The mixture was concentrated under reduced pressure and the residue was partitioned between DCM (100 mL) and water (50 mL). The aqueous layer was then separated with DCM (50 mL). 2) Extraction was performed, and the combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid chromatography (80 g silica column, DCM / MeOH, where MeOH ranged from 0–10%) to provide the desired product (6.0 g, white solid, yield: 90.7%). LC-MS: (ESI) m / z (M+Na), 624.2.

[1412] Step 6b. At 20°C and under H2, add Pd / C (80 mg, 0.75 mmol) in CH3OH (30 mL) to a solution of the compound from Step 6a (1.5 g, 2.49 mmol). Stir the mixture for 12 hours. Concentrate the mixture to obtain the desired compound, which can be used in the next step without purification (1.1 g, white solid, yield: 86.35%). LC-MS: (ESI) m / z (M+Na), 534.2.

[1413] Step 6c. At 30°C, acetic acid (0.70 g, 11.7 mmol) was added in a single addition to a solution of the compound from Step 6b (1.5 g, 2.93 mmol), copper bis(acetic acid) hydrate (0.18 g, 0.88 mmol), and Pb(CH3COO)4 (4.69 g, 4.39 mmol) in THF (50 mL). The mixture was stirred at 30°C for 2 hours. The mixture was concentrated under reduced pressure and the residue was partitioned between DCM (100 mL) and water (50 mL). The aqueous layer was separated with DCM (50 mL). 2) Extraction was performed, and the combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid chromatography (45 g silica column, PE / EA, where EA ranged from 0-55%) to provide the desired product (1.1 g, white solid, yield: 71.4%). LC-MS: (ESI) m / z (M+Na), 548.4.

[1414] Step 6d. Add 4-methylbenzenesulfonic acid (0.04 g, 0.21 mmol) to a mixture of the compound from step 6c (1.1 g, 2.09 mmol) and benzyl glycolate (1.74 g, 10.46 mmol) in THF (20 mL). Stir the mixture at 25 °C for 2 hours. Concentrate the mixture under reduced pressure and use DCM (50 mL). 2) Extraction was performed, and the combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid chromatography (30 g silica column, PE / EA, where EA ranged from 0-60%) to provide the desired product (0.49 g, white solid, yield: 37.1%). LC-MS: (ESI) m / z (M+Na), 654.2.

[1415] Step 6e. At 20 °C and under H2, Pd / C (20 mg, 0.23 mmol) was added to a mixture of the compound from step 6d (0.49 g, 0.78 mmol) in MeOH (20 mL). The resulting mixture was then stirred at 20 °C for 2 hours. The solution was filtered and concentrated to obtain the desired compound (0.45 g, white solid, quantitative yield), which could be used in the next step without purification. LC-MS: (ESI) m / z (M+Na), 564.3.

[1416] Step 6f. At 0°C, add DIPEA (200 mg, 1.55 mmol) in 10 mL of DMF to a solution of the compound from Step 6e (280 mg, 0.52 mmol), (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,10,12,13,15-octahydrocyclohexane[1,2,3-de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-10,13-dione methanesulfonic acid (CAS 171335-80-1) (250 mg, 0.46 mmol), and DMTMM (250 mg, 0.52 mmol) for 5 minutes. Allow the mixture to warm naturally to 20°C and stir for 2 hours. Dilute the aqueous layer with DCM (50 mL). 2) Extracted with H2O (50 mL), the combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid chromatography (10 g silica column, DCM / MeOH, where MeOH ranged from 0–10%) to provide the desired product (0.27 g, brown solid, yield: 54.5%). LC-MS: (ESI) m / z (M+H), 960.2.

[1417] Step 6g. Diethylamine (0.08 mL, 0.78 mmol) in DMF (2 mL) was added to a solution of the compound (50 mg, 0.05 mmol) from Step 6f at 0 °C for 2 hours. The solution was filtered and concentrated to obtain the desired compound (42 mg, white oil, quantitative yield), which could be used in the next step without purification. LC-MS: (ESI) m / z (M+H), 737.2.

[1418] Step 6h. Add the compound from step 6g (42 mg, 0.057 mmol) to a mixture of the compound from step 3h (100 mg, 0.22 mmol), DIPEA (22.1 mg, 0.17 mmol), and DMF (5 mL). Stir the mixture at 0°C for 2 hours. Remove the solvent under vacuum and purify the residue by preparative HPLC (C18, H2O (containing 0.1% formic acid) in 20%–60% acetonitrile) to provide the title compound as a yellow solid (7.1 mg, 11.6% yield). LC-MS: (ESI) m / z (M+H), 1074.4. 1H NMR (400 MHz, DMSO-d6) δ 8.80-8.70 (m, 1H), 8.51(d, J = 8.4 Hz, 1H), 8.30-8.13 (m, 2H), 7.90 (d, J = 7.6 Hz, 1H), 7.79 (d, J= 10.8 Hz, 1H), 7.32 (s, 1H), 6.99 (s, 2H), 6.80-6.75 (m, 1H), 6.59-6.51 (m,1H), 5.65-5.59 (m, 1H), 5.42 (s, 2H), 5.21 (s, 2H), 4.65-4.55 (m, 2H), 4.28 –4.10 (m, 3H), 3.99 (s, 2H), 3.75-3.71 (m, 2H), 3.58-3.55 (m, 8H), 2.90-2.86(m, 2H), 2.40 (s, 3H), 2.20-2.18 (m, 2H), 2.16 – 2.10 (m, 4H), 1.90-1.85 (m, 2H), 1.65-1.61 (m, 2H), 1.50-1.46 (m, 4H), 1.35 (s, 9H), 0.87 (t, J = 7.2 Hz, 3H).

[1419] Example 7: Preparation of LP7 (MC-SGE-DXD)

[1420]

[1421] (4S)-4-{[(5S)-12-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-5-(hydroxymethyl)-1,4,7-trioxo-3,6-diazadodecane-1-yl]amino}-5-({[(2-{[(1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,12,15-hexahydro-1H-cyclohexane[1,2,3-de]pyrano[3',4':6,7]inzizo[1,2-b]quinolino-1-yl]amino}-2-oxoethyl)oxy]methyl}amino)-5-oxopentanoic acid

[1422]

[1423] Step 7a. At 20°C, DIEA (4.27 g, 33.0 mmol) was added in a single dose to a solution of (2S)-2-({[(9H-fluorene-9-ylmethyl)oxy]carbonyl}amino)-5-[(2-methylprop-2-yl)oxy]-5-oxovalerate (5000 mg, 11.751 mmol), benzyl aminoacetate (1941.22 mg, 11.751 mmol), EDCI (2703.30 mg, 14.102 mmol), and HOBt (1905.57 mg, 14.102 mmol) in DCM (150 mL). The mixture was stirred at 20°C for 16 hours. The mixture was concentrated under reduced pressure and the residue was partitioned between DCM (100 mL) and water (50 mL). The aqueous layer was separated with DCM (50 mL). 2) Extraction was performed, and the combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid chromatography (80 g silica gel column, DCM / MeOH, where MeOH ranged from 0–10%) to provide the desired product as a yellow solid (5500 mg, 9.604 mmol, 81.73%). LC-MS: (ESI) m / z (M-55), 517.3.

[1424] Step 7b. At 20°C and under H2, add 10% Pd / C (37.17 mg, 0.349 mmol) in MeOH (20 mL) to the mixture from Step 7a (2000 mg, 3.492 mmol). Stir the mixture for 12 hours. Concentrate the mixture to obtain the desired compound (1600 mg, 2.487 mmol, 71.21%) as a yellow solid, which can be used in the next step without purification. LC-MS: (ESI) m / z (M-55), 427.2.

[1425] Step 7c. At 30°C, pyridine (0.367 mL, 4.559 mmol) was added in a single step to a solution of the compound from step 7b (1100 mg, 2.280 mmol), copper bis(acetic acid) hydrate (227.57 mg, 1.140 mmol), lead tetraacetate (1516.13 mg, 3.419 mmol), in THF (15 mL), and toluene (15 mL). The mixture was stirred at 30°C for 16 hours. The reaction mixture was concentrated under vacuum and purified by rapid chromatography (PE / EA, where EA ranged from 0–40%) to provide the desired compound as a white solid (280 mg, 0.564 mmol, 24.7%). LC-MS: (ESI) m / z (M+Na), 519.2.

[1426] Step 7d. A solution of the compound from step 7c (280 mg, 0.564 mmol), benzyl glycolate (468.53 mg, 2.819 mmol), and 4-methylbenzenesulfonic acid (9.71 mg, 0.056 mmol) in THF (20 mL) was stirred at 30 °C for 16 h. The reaction mixture was concentrated under vacuum and purified by rapid chromatography (PE / EA, where EA ranged from 0–50%) to provide the desired compound (280 mg, 0.465 mmol, 82.39%). LC-MS: (ESI) m / z (M+Na), 625.4.

[1427] Step 7e. At 30°C and under H2, 10% Pd / C (280 mg, 0.465 mmol) in MeOH (15 mL) was added to a solution of the compound from step 7d over 16 hours. The mixture was concentrated to obtain the desired compound (230 mg, 96.6% yield) as a yellow solid, which could be used in the next step without purification. LC-MS: (ESI) m / z (M+Na), 535.2.

[1428] Step 7f. Add DMTMM (159 mg, 0.54 mmol) to the mixture of the compound from Step 7e (0.23 g, 0.49 mmol), eczemab mesylate (CAS: 169869-90-3) (190 mg, 0.36 mmol), and DIPEA (174 mg, 1.35 mmol) in DMF (10 mL). Stir the mixture at 0 °C for 2 hours. Concentrate the mixture under reduced pressure and partition the residue between DCM (20 mL) and water (20 mL). Dilute the aqueous layer with DCM (20 mL) 2) Extraction was performed, and the combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid chromatography (10 g silica column, DCM / CH3OH, where CH3OH ranged from 0–5%) to provide the desired product (200 mg, white solid, yield: 47.9%). LC-MS: (ESI) m / z (M+H), 930.2.

[1429] Step 7g. At 0°C, TFA (0.32 mL, 4.3 mmol) was added to a solution of the compound from Step 7f (200 mg, 0.21 mmol) in DCM (1.5 mL). After stirring at 25°C for 3 hours, the solution was concentrated, and the residue was purified by rapid chromatography (10 g silica column, DCM / CH3OH, where CH3OH ranged from 0–5%) to provide the desired product (15 mg, yellow solid, yield: 7.9%). LC-MS: (ESI) m / z (M+H), 946.2.

[1430] Step 7h. Diethylamine (18.8 mg, 0.25 mmol) was added to a solution of the compound from step 7g (15 mg, 0.017 mmol) in DMF (0.5 mL) at 0 °C. After stirring at 0 °C for 2 hours, the solution was filtered and concentrated to obtain the desired compound (11 mg, yellow oil, quantitative yield), which could be used in the next step without purification. LC-MS: (ESI) m / z (M+H), 652.2.

[1431] Step 7i. At 0 °C, DIEA (6.55 mg, 0.05 mmol) was added to a solution of the compound from step 7h (11 mg, 0.017 mmol) and the compound from step 3h (7.64 mg, 0.017 mmol) in DMF (0.5 mL). After stirring at 25 °C for 2 hours, the solvent was removed under vacuum and the residue was purified by preparative HPLC (C18, 20%–60% acetonitrile in H2O (containing 0.1% FA)) to provide the title compound (3.2 mg, 19% yield) as a white solid. LC-MS: (ESI) m / z (M+H), 989.5. 1H NMR (400 MHz, DMSO) δ12.15 (s, 1H), 8.73 – 8.69 (m,1H), 8.55 – 8.51 (m, 1H), 8.20 – 8.16 (m, 1H), 8.04 – 7.95 (m, 2H), 7.81 –7.79 (m, 1H), 7.31 (s, 1H), 6.99 (s, 2H), 6.51 (s, 1H), 5.64 – 5.60 (m, 1H), 5.42 (s, 2H), 5.22 (s, 2H), 4.65 – 4.61 (m, 2H), 4.23 – 4.18 (m, 2H), 4.00(s, 2H), 3.73 – 3.70 (m, 1H), 3.58 – 3.55 (m, 2H), 3.20 – 3.14 (m, 2H), 2.40(s, 3H), 2.23 – 2.13 (m, 8H), 1.91 – 1.87 (m, 4H), 1.48 – 1.43 (m, 4H), 1.27– 1.20 (m, 4H), 0.87 (t, J = 7.2 Hz, 3H).

[1432] Example 8: Preparation of LP8 (MC-SGC(PEG)-DXD)

[1433]

[1434] 6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-N-((9S,15S)-9-(((2-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl)amino)-2-oxoethoxy)methyl)carbamoyl)-1,16-dihydroxy-4,11,14-trioxo-3,5,10,13-tetraazahexadecane-15-yl)hexanoamide

[1435]

[1436] Step 8a. A mixture of 2-{[(2-methylpropyl-2-yl)diphenylsilyl]oxy}eth-1-amine (5 g, 16.69 mmol) and 1-{[(2,5-dioxotetrahydro-1H-pyrrolo-1-yl)oxy]carbonyl}oxy)tetrahydropyrrolo-2,5-dione (4.28 g, 16.69 mmol) in DCM (50 mL) was stirred at 25 °C for 2 hours. The solution was filtered and concentrated to obtain the desired compound (5 g, colorless oil, yield: 67.9%), which was ready for use in the next step without further purification. LC-MS: (ESI) m / z (M+Na), 463.2.

[1437] Step 8b. A mixture of the compound from step 8a (5 g, 11.34 mmol), (2S)-5-amino-2-({[(9H-fluorene-9-ylmethyl)oxy]carbonyl}amino)valeric acid (3.42 g, 9.67 mmol), and DIPEA (4.40 g, 34.0 mmol) in DMF (50 mL) was stirred at 25 °C for 18 hours. The mixture was concentrated, and the residue was purified by reversed-phase column chromatography (H2O / CH3CN, where CH3CN ranged from 0–50%) to provide the desired product (0.8 g, white solid, yield: 10.4%). LC-MS: (ESI) m / z (M+H), 680.2.

[1438] Step 8c. The mixture of the compound from step 8b (0.8 g, 1.17 mmol), benzyl aminoacetate hydrochloride (0.31 g, 1.53 mmol), EDCI (0.29 g, 1.53 mmol), HOBT (0.21 g, 1.53 mmol), and DIEA (0.46 g, 3.53 mmol) in DCM (20 mL) was stirred at 25 °C for 18 hours. The reaction was quenched with water (20 mL). The aqueous layer was separated and treated with DCM (20 mL). 2) Extraction. The combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (PE / EA, where EA ranged from 0-50%) to provide the desired product (0.5 g, white solid, yield: 51.4%). LC-MS: (ESI) m / z (M+H), 827.6.

[1439] Step 8d. The mixture of the compound from step 8c (500 mg, 4.69 mmol) and Pd / C (100 mg, 0.03 mmol) in CH3OH (15 mL) was stirred at 25 °C under a H2 atmosphere for 18 hours. The mixture was filtered and concentrated to obtain the desired compound (0.4 g, white solid, yield: 89.9%), which could be used in the next step without further purification. LC-MS: (ESI) m / z (M+H), 737.4.

[1440] Step 8e. The mixture of the compound from step 8d (400 mg, 0.54 mmol), acetic acid (130 mg, 2.17 mmol), acetic acid (32.5 mg, 0.16 mmol), and Pb(OAc)4 (721 mg, 1.6 mmol) in THF (15 mL) was stirred at 30 °C for 2 hours. The mixture was diluted with DCM (20 mL) and washed with water (10 mL). The aqueous layer was separated and washed with DCM (20 mL). 4) Extraction. The combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (PE / EA, where EA ranged from 0-50%) to provide the desired product (180 mg, yellow solid, yield: 44.16%). LC-MS: (ESI) m / z (M+Na), 773.4.

[1441] Step 8f. The mixture of the compound from step 8e (180 mg, 0.24 mmol), benzyl glycolate (199 mg, 1.20 mmol), and 4-methylbenzenesulfonic acid hydrate (23 mg, 0.12 mmol) in THF (5 mL) was stirred at 0 °C for 2 hours. The mixture was diluted with DCM (20 mL) and washed with water (10 mL). The aqueous layer was separated and washed with DCM (20 mL). 4) Extraction. The combined organic phases were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (PE / EA, where EA ranged from 0-60%) to provide the desired product (60 mg, white solid, 29.2% yield). LC-MS: (ESI) m / z (M+Na), 857.6.

[1442] Step 8g. The mixture of the compound from Step 8f (60 mg, 0.07 mmol) and Pd / C (20 mg, 0.19 mmol) in MeOH (8 mL) was stirred at 25 °C under a H2 atmosphere for 18 hours. The solution was filtered and concentrated to obtain the desired compound (50 mg, white solid, 93.1% yield), which could be used in the next step without further purification. LC-MS: (ESI) m / z (M+H), 767.4.

[1443] Step 8h. At 0°C, DMTMM (46 mg, 0.15 mmol) and DIEA (30 mg, 0.23 mmol) were added to a mixture of the compound from step 8g (60 mg, 0.08 mmol), eczemab mesylate (CAS: 169869-90-3) (29 mg, 0.07 mmol) in DMF (8 mL). The solution was diluted with DCM (20 mL) and washed with water (10 mL), and the phases were separated. The aqueous layer was extracted four times with DCM (20 mL), and the combined organic phases were dried over sodium sulfate and the solvent was removed under reduced pressure. The residue was purified by reversed-phase column chromatography (H2O / CH3CN, where CH3CN ranged from 0-60%) to provide the desired product (30 mg, yellow solid, yield: 32.6%). LC-MS: (ESI) m / z (1 / 2M+H), 593.0.

[1444] Step 8i. The mixture of the compound from step 8h (25 mg, 0.02 mmol) and fluoropyridine (5 mg, 0.04 mmol) in pyridine (2 mL) was stirred at 25 °C for 18 h. The solution was concentrated, and the residue was purified by preparative HPLC (C18, 20%–60% acetonitrile in H2O (containing 0.1% FA)) to provide the desired product (10 mg, yellow solid, yield: 50.1%). LC-MS: (ESI) m / z (M+H), 946.2.

[1445] Step 8j. The mixture of the compound from Step 8i (10 mg, 0.01 mmol) and diethylamine (15 mg, 0.21 mmol) in DMF (0.5 mL) was stirred at 0 °C for 2 hours. The solution was filtered and concentrated to obtain the desired compound, which could be used in the next step without purification (8 mg, colorless oil, quantitative yield). LC-MS: (ESI) m / z (M+H), 724.4.

[1446] Step 8k. A mixture of the compound from Step 8j (8 mg, 0.01 mmol), the compound from Step 3h (5 mg, 0.01 mmol), and DIEA (4.3 mg, 0.03 mmol) in DMF (0.5 mL) was stirred at 0 °C for 2 h. The solvent was removed under vacuum, and the residue was purified by preparative HPLC (C18, 20%–60% acetonitrile in H2O (containing 0.1% FA)) to provide the title compound (4.0 mg, 34.1% yield) as a white solid. LC-MS: (ESI) m / z (1 / 2 M + H), 531.4. 1 H NMR (400 MHz, DMSO) δ 8.74 – 8.71 (m, 1H), 8.55 – 8.51 (m, 1H), 8.20 – 8.14 (m, 1H), 7.95 – 7.91 (m, 2H), 7.82 – 7.79 (m, 1H), 7.32 (s, 1H), 6.99(s, 2H), 6.52 (s, 1H), 5.95 – 5.93 (s, 1H), 5.84 – 5.82 (s, 1H), 5.58 – 5.55(m, 1H), 5.43 (s, 2H), 5.21 (s, 2H), 4.98 – 4.95 (m, 1H), 4.69 – 4.63 (m,3H), 4.26 – 4.20 (m, 2H), 3.99 (s, 2H), 3.75 – 3.71 (m, 2H), 3.58 – 3.54 (m,3H), 3.17 – 3.15 (m, 1H), 3.05 – 3.01 (m, 2H), 2.95 – 2.92 (m, 2H), 2.40 (s,3H), 2.20 – 2.10 (m, 4H), 1.91 – 1.84 (m, 2H), 1.70 – 1.62 (m, 2H), 1.52 –1.46 (m, 6H), 1.39 – 1.25 (m, 6H), 0.87 (t, J = 7.2 Hz, 3H).

[1447] The following examples 9-11 were prepared using a procedure similar to that described above:

[1448]

[1449] LP9 (MC-TSR-DXD)

[1450] LP10 (MC-SGO-DXD)

[1451] LP11 (MC-SGK-DXD)

[1452] Example 12: Preparation of LP12 (MsP-SGC-DXD)

[1453]

[1454] N-((7S,13S)-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl)amino)-14-hydroxy-1,6,9,12-tetraoxo-7-(3-ureopropyl)-3-oxa-5,8,11-triazatetradecane-13-yl)-6-(2-(methylsulfonyl)pyrimidin-5-yl)hex-5-yneamide

[1455]

[1456] Step 12a. At 0 °C, HATU (1.49 g, 3.912 mmol), 2-methylpropyl-2-ylaminoacetic acid (0.41 g, 3.130 mmol), and DIPEA (1.293 mL, 7.824 mmol) were slowly added to a solution of (2S)-2-({[(9H-fluorene-9-ylmethyl)oxy]carbonyl}amino)-3-[(2-methylprop-2-yl)oxy]propionic acid (1 g, 2.608 mmol) in DMF (20 mL). The resulting mixture was stirred at 15 °C for about 16 h. It was concentrated under reduced pressure and the residue was partitioned between EA (50 mL) and water (20 mL). The aqueous layer was extracted with EA (20 mL × 2), and the combined organic matter was dried (Na2SO4), filtered, and concentrated. The residue was purified by chromatography (silica, 0-50% PE in EA) to provide the desired compound (1.1 g, white solid, 84% yield). LC-MS: (ESI) m / z (M+Na), 519.4.

[1457] Step 12b. At 0 °C, TFA (10 mL, 2.215 mmol) was slowly added to a solution of the compound from Step 12a (3.5 g, 7.048 mmol) in DCM (50 mL). The resulting mixture was stirred at 15 °C for approximately 16 h. It was concentrated under reduced pressure and the residue was purified by RP-FC (C18, 5%–95% MeCN in H2O) to provide the desired compound (2.2 g, white solid, 81% yield). LC-MS: (ESI) m / z (M+1), 385.4.

[1458] Step 12c. At 0 °C, HATU (113.28 mg, 0.298 mmol), DIPEA (0.098 mL, 0.596 mmol), and the compound from step 3g (135 mg, 0.199 mmol) were slowly added to a solution of the compound from step 12b (91.61 mg, 0.238 mmol) in DMF (5 mL). The resulting mixture was stirred at 20 °C for about 4 h. It was concentrated under reduced pressure and the residue was partitioned between EA (20 mL) and water (10 mL). The aqueous layer was extracted with EA (10 mL × 2), and the combined organic matter was dried (Na2SO4), filtered, and concentrated. The residue was purified by chromatography (0-10% DCM in silica, MeOH) to provide the desired compound (100 mg, white solid, 48% yield). LC-MS: (ESI) m / z (M+H), 1047.2.

[1459] Step 12d. Diethylamine (0.045 mL, 0.430 mmol) was slowly added to a solution of the compound from step 12c (150 mg, 0.143 mmol) in DMF (5 mL) at 0 °C. The resulting mixture was stirred at 20 °C for about 2 h. It was concentrated under reduced pressure, and the mixture was concentrated to obtain the desired compound, which could be used in the next step without purification (118 mg, pale yellow oil, quantitative yield). LC-MS: (ESI) m / z (M+H), 824.2.

[1460] Step 12e. At 0 °C, HATU (83.08 mg, 0.218 mmol), DIPEA (0.072 mL, 0.437 mmol), and the compound from Step 12d (120 mg, 0.146 mmol) were slowly added to a solution of 6-[2-(methyldioxo-λ6-thioalkyl)pyrimidin-5-yl]hex-5-ynyleneic acid (CAS2356229-58-6) (58.62 mg, 0.218 mmol) in DMF (5 mL). The resulting mixture was stirred at 20 °C for about 4 h. It was concentrated under reduced pressure and the residue was purified by preparative HPLC (C18, 20%–60% acetonitrile in H2O (containing 0.1% formic acid)) to provide the title compound (30.6 mg, yellow solid, 19% yield). LC-MS: (ESI) m / z (M+H), 1075.2. 1 H NMR (400 MHz, DMSO) δ 9.11 (s, 2H), 8.74 (t, J = 6.4 Hz, 1H), 8.52 (d, J = 8.8 Hz, 1H), 8.21 (t, J = 5.6 Hz, 1H), 8.05 (d, J = 7.2 Hz, 1H), 7.94 (d, J = 7.6 Hz, 1H), 7.79-7.75 (m, 1H), 7.31(s, 1H), 5.95 (s, 1H), 5.61-5.58 (m, 1H), 5.43 (s, 2H), 5.20 (s, 2H), 4.63-4.61 (m, 2H), 4.27-4.16 (m, 3H), 4.00 (s, 3H), 3.74 - 3.71 (m, 3H), 3.59-3.57(m, 3H), 3.25-3.11 (m, 2H), 2.95-2.89 (m, 2H), 2.58-2.54 (m, 2H), 2.53-2.51(m, 3H), 2.38-2.33 (m, 5H), 2.23- 2.13 (m, 2H), 1.92-1.77 (m, 4H), 1.71 –1.62 (m, 1H), 1.52-1.45 (m, 1H), 1.40-1.25 (m, 2H), 0.85 (t, J = 7.2 Hz, 3H).

[1461] Example 13: Preparation of LP13 (MsP-SGR-DXD)

[1462]

[1463] N-((7S,13S)-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl)amino)-7-(3-guanidinopropyl)-14-hydroxy-1,6,9,12-tetraoxo-3-oxa-5,8,11-triazatetradecane-13-yl)-6-(2-(methylsulfonyl)pyrimidin-5-yl)hex-5-yneamide

[1464]

[1465] Step 13a. The mixture of the compound from step 12b and the compound from step 1g (80 mg, 0.12 mmol), DIEA (45 mg, 0.35 mmol), and DMTMM (70 mg, 0.24 mmol) in DMF (2 mL) was stirred at 25 °C for 18 hours. The mixture was concentrated under vacuum, and the residue was purified by reversed-phase column chromatography (C18, H2O / CH3CN, where CH3CN ranged from 0–50%) to provide the desired product (40 mg, 32% yield) as a yellow solid. LC-MS: (ESI) m / z (M+H), 1044.6.

[1466] Step 13b. The mixture of the compound from step 13a (40 mg, 0.038 mmol) and DEA (79 μL, 0.76 mmol) in DMF (2 mL) was stirred at 0 °C for 2 hours. The solution was filtered and concentrated under vacuum to obtain the desired compound (32 mg) as a brown solid, which could be used in the next step without purification. LC-MS: (ESI) m / z(M+H), 822.7.

[1467] Step 13c. A mixture of the compound from step 13b (32 mg, 0.039 mmol), 6-(2-(methanesulfonyl)pyrimidin-5-yl)hex-5-alkynic acid (CAS: 2356229-58-6) (12 mg, 0.043 mmol), HATU (29 mg, 0.078 mmol), and DIEA (15 mg, 0.117 mmol) in DMF (3 mL) was stirred at 25 °C for 2 hours. The mixture was concentrated under vacuum, and the residue was purified by preparative HPLC (C18, 20%–60% acetonitrile in H2O (containing 0.1% TFA)) to provide the title compound (9.1 mg, 23% yield) as a yellow solid. LC-MS: (ESI) m / z (M+H), 1073.2. 1 HNMR (400 MHz, DMSO) δ 9.11 (s, 2H), 8.77 – 8.72 (m, 1H), 8.56 – 8.54 (m, 1H), 8.25 – 8.21 (m, 1H), 8.12 – 8.06 (m 1H), 8.03 – 7.99 (m, 1H), 7.83 – 7.80 (m,1H), 7.46 – 7.42 (m, 1H), 7.33 (s, 1H), 7.11 – 6.78 (m, 2H), 6.55 (s, 1H),5.63 – 5.59 (m, 1H), 5.43 (s, 2H), 5.21 (s, 2H), 5.10 – 4.99 (m, 1H), 4.70 –4.64 (m, 2H), 4.29 – 4.25 (m, 2H), 4.01 (s, 2H), 3.80 – 3.74 (m, 2H), 3.66 –3.60 (m, 2H), 3.41 (s, 3H), 3.22 – 3.19 (m, 2H), 3.15 – 3.07 (m, 3H), 2.60 –2.58 (m, 2H), 2.40 (s, 3H), 2.39 – 2.37 (m, 2H), 2.23 – 2.18 (m, 2H), 1.88 –1.79 (m, 4H), 1.51 – 1.46 (m, 4H), 0.88 (t, J = 7.2 Hz, 3H).

[1468] The following examples 14-18 were prepared using a procedure similar to that described above:

[1469]

[1470]

[1471] LP14 (MsP-SGE-DXD)

[1472] LP15 (MsP-TSR-DXD)

[1473] LP16 (MsP-SGK-DXD)

[1474] LP17 (MsP-YSR-DXD)

[1475] LP18 (MsP-Y(Me)SR-DXD)

[1476] Example 19: Preparation of LP19 (MC-SGC-PABC-Ecinotecan)

[1477]

[1478] 4-((S)-2-(2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)hexamido)-3-hydroxypropamido)acetamyl)-5-ureidopentamido)benzyl((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl)carbamate

[1479]

[1480] Step 19a. At 0 °C, EEDQ (5.90 g, 23.88 mmol) and (4-aminophenyl)methanol (2.94 g, 23.88 mmol) were slowly added to a solution of (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-5-ureidovalerate (10.0 g, 19.9 mmol) in DCM (100 mL). The resulting mixture was stirred at 15 °C for about 16 hours. It was concentrated under reduced pressure and the residue was partitioned between EA (300 mL) and water (100 mL). The aqueous layer was extracted with EA (200 mL × 2), and the combined organic matter was dried (Na2SO4), filtered, and concentrated. The residue was purified by chromatography (0-10% DCM in silica, MeOH) to provide the desired compound (7.0 g, white solid, 70% yield). LC-MS: (ESI) m / z (M+H), 503.2.

[1481] Step 19b. At 0 °C, 4-nitrophenyl carbonate (510 mg, 2.38 mmol) and pyridine (310 mg, 3.98 mmol) were slowly added to a solution of the compound from Step 19a (1.0 g, 1.99 mmol) in DMF (20 mL). The resulting mixture was stirred at 15 °C for about 16 h. It was concentrated under reduced pressure and the residue was partitioned between EA (50 mL) and water (20 mL). The aqueous layer was extracted with EA (30 mL × 2), and the combined organic matter was dried (Na2SO4), filtered, and concentrated. The residue was purified by chromatography (0-10% DCM in silica, MeOH) to provide the desired compound (270 mg, white solid, 20% yield). LC-MS: (ESI) m / z (M+H), 668.2.

[1482] Step 19c. At 0 °C, (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,10,12,13,15-octahydrocyclohexane[1,2,3-de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-10,13-dione (107.61 mg, 0.247 mmol) and DIPEA (0.111 mL, 0.674 mmol) were slowly added to a solution of the compound from Step 19b (150 mg, 0.225 mmol) in DMF (5 mL). The resulting mixture was stirred at 15 °C for about 3 h. It was concentrated under reduced pressure and the residue was purified by RP-FC (5%-95% MeCN in C18, H2O) to provide the desired compound (120 mg, yellow solid, 55% yield). LC-MS: (ESI) m / z (M+1), 964.4.

[1483] Step 19d. Diethylamine (27.2 mg, 0.372 mmol) was slowly added to a solution of the compound from step 19c (120 mg, 0.124 mmol) in DMF (0.5 mL) at 0 °C. The resulting mixture was stirred at 15 °C for about 1 h. The mixture was concentrated to obtain the desired compound, which could be used in the next step without purification (90 mg, pale yellow solid, quantitative yield). LC-MS: (ESI) m / z (M+1), 742.4.

[1484] Step 19e. At 0 °C, the compound from step 3 h (109.784 mg, 0.242 mmol) and DIPEA (46.917 mg, 0.363 mmol) were slowly added to a solution of the compound from step 19 d (90 mg, 0.121 mmol) in DMF (0.5 mL). The resulting mixture was stirred at 15 °C for about 3 h. It was concentrated under reduced pressure and the residue was purified by preparative HPLC (C18, 20%–60% acetonitrile in H2O (containing 0.1% formic acid)) to provide the title compound (10.3 mg, yellow solid, 7.8% yield). LC-MS: (ESI) m / z (M+H), 1079.4. 1H NMR (400 MHz, DMSO) δ 9.87 (s,1H), 8.14 (m, 1H), 7.99 (d, J = 8.8 Hz, 1H), 7.93 (d, J = 7.6 Hz, 1H), 7.86(d, J = 7.2 Hz, 1H), 7.71 (d, J = 10.8 Hz, 1H), 7.56 (m, 2H), 7.30 (d, J =8.4 Hz, 2H), 7.25 (s, 1H), 6.92 (s, 2H), 6.56-6.30 (m, 1H), 5.90 (s, 1H),5.38 (s, 2H), 5.22 (s, 3H), 5.01 (s, 2H), 4.33 (m, 1H), 4.19-4.14 (m, 1H), 3.68-3.67 (m, 2H), 3.54-3.49 (m, 3H), 3.10-2.99 (m, 2H), 2.82-2.88 (m, 4H), 2.60-2.58 (m, 1H), 2.31 (s, 3H), 2.28-2.24 (m, 1H), 2.08-2.05 (m, 3H), 1.86-1.76 (m, 2H), 1.66-1.59 (m, 1H), 1.54-1.52 (m, 1H), 1.49-1.36(m, 6H), 1.17-1.08 (m, 3H), 0.81 (t, J = 7.2 Hz, 3H).

[1485] Example 20: Preparation of LP20 (MC-SGO(dEt)-PABC-Ecinotecan)

[1486]

[1487] 4-((S)-5-(diethylamino)-2-(2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)hexamido)-3-hydroxypropamido)acetamyl)pentamido)benzyl((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-1,2,3,9,10,12,13,15-octahydrobenzo[de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl)carbamate

[1488]

[1489] Step 20a. At 20 °C, sodium cyanoborohydride (3.55 mL, 56.4 mmol) in EtOH (50 mL) was added to a solution of (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-5-aminopentanoic acid (5 g, 14.1 mmol), acetaldehyde (4.97 g, 112.8 mmol), and acetic acid (2.54 g, 42.3 mmol) over 4 hours. The solution was filtered and the filtrate was purified by passing it through a reversed-phase column (120 g C18 gel column, H2O / CH3CN, where CH3CN ranged from 0–40%) to provide the desired product (2 g, white solid, yield: 34.5%). LC-MS: (ESI) m / z (M+H), 411.2.

[1490] Step 20b. The compound from Step 20a (0.44 g, 5.36 mmol) was dissolved in DMF (15 mL) at 25 °C for 18 hours. The mixture was concentrated under reduced pressure, and the residue was partitioned between DCM (100 mL) and water (50 mL). The aqueous layer was then separated with DCM (50 mL). 2) Extraction was performed, and the combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid chromatography (30 g silica column, DCM / MeOH, where MeOH ranged from 0–10%) to provide the desired product (0.4 g, white solid, yield: 31.8%). LC-MS: (ESI) m / z (M+H), 516.4.

[1491] Step 20c. Add DIPEA (300 mg, 2.33 mmol) to a mixture of the compound from Step 20b (0.4 mg, 0.77 mmol), 4-nitrophenyl[(4-nitrophenyl)oxy]carbamate (0.47 mg, 1.55 mmol), and DMAP (10 mg, 0.08 mmol) in DMF (8 mL). Stir the mixture at 0 °C for 2 hours. Filter the solution and purify the filtrate by passing it through a reversed-phase column (40 g C18 gel column, H2O / CH3CN, where CH3CN ranges from 0–50%) to provide the desired product (200 mg, yellow solid, yield: 37.8%). LC-MS: (ESI) m / z (M+H), 681.2.

[1492] Step 20d. DIPEA (85 mg, 0.66 mmol) was added to a mixture of (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,10,12,13,15-octahydrocyclohexane[1,2,3-de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-10,13-dione methanesulfonic acid (140 mg, 0.26 mmol) and the compound from step 20c (150 mg, 0.22 mmol) in DMF (5 mL). The mixture was stirred at 25 °C for 2 hours. The solution was filtered and the filtrate was purified by passing it through a reversed-phase column (40 g C18 gel column, H2O / CH3CN, where CH3CN ranged from 0-80%) to provide the desired product (110 mg, yellow solid, yield: 51.1%). LC-MS: (ESI) m / z (M+H), 978.2.

[1493] Step 20e. Diethylamine (60 mg, 0.76 mmol) was added to a mixture of the compound from step 20d (50 mg, 0.05 mmol) in DMF (0.5 mL) at 0 °C. The resulting mixture was then stirred at 0 °C for 2 hours. The solution was filtered and concentrated to obtain the desired compound (35 mg, white solid, yield: 90.6%), which was ready for use in the next step without purification. LC-MS: (ESI) m / z (M+H), 755.4.

[1494] Step 20f. Add the compound from step 3h (40 mg, 0.09 mmol) to a mixture of the compound from step 20e (35 mg, 0.046 mmol) and DIPEA (0.023 mL, 0.14 mmol) in DMF (1 mL). Stir the mixture at 20 °C for 2 hours. Remove the solvent under vacuum and purify the residue by preparative HPLC (C18, 20%–60% acetonitrile in H2O (containing 0.1% TFA)) to provide the title compound as a yellow solid (17.7 mg, 34.9% yield). LC-MS: (ESI) m / z (M+H), 1093.6. 1H NMR (400 MHz, DMSO) δ 9.98 (s, 1H), 8.96 (s,1H), 8.30 - 8.25 (m, 1H), 8.06 (d, J = 8.0 Hz, 1H), 8.01 - 7.97 (m, 1H), 7.81- 7.79 (m, 1H), 7.63 (d, J = 8.0 Hz, 2H), 7.40 (s, 2H), 7.32 (s, 1H), 7.00(s, 2H), 6.50 (m, 1H), 5.45 (s, 2H), 5.35 - 5.29 (m, 3H), 5.10 (s, 2H), 4.50- 4.47 (m, 1H), 4.27 - 4.18 (m, 1H), 3.81 - 3.77 (m, 2H), 3.65 - 3.60 (m,2H), 3.32 - 3.20 (m, 4H), 3.15 - 3.09 (m, 8H), 2.39 (s, 3H), 2.25 - 2.11 (m,5H), 1.98 -1.75 (m, 4H), 1.70 - 1.67 (m, 2H), 1.50 - 1.48 (m, 4H), 1.18 (t, J= 6.8 Hz, 6H), 0.89 (t, J = 7.2 Hz, 3H).

[1495] Example 21: Preparation of LP21 (MC-SGE-PABC-Ecinotecan)

[1496]

[1497] (S)-4-(2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)hexamido)-3-hydroxypropamido)acetamido)-5-((4-(((((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-1,2,3,9,10,12,13,15-octahydrobenzo[de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl)carbamoyl)oxy)methyl)phenyl)amino)5-oxopentanoic acid

[1498] LP21 was prepared according to step 21 of Example 22.

[1499] Example 22: LP22 (MC-SGE) t Preparation of Bu)-PABC-Ecinotecan

[1500]

[1501] (4-{[(2S,8S)-15-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)-8-(hydroxymethyl)-2-{3-[(2-methylpropyl-2-yl)oxy]-3-oxopropyl}-1,4,7,10-tetraoxo-3,6,9-triazapentadecan-1-yl]amino}phenyl)methyl{[(1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,12,15-hexahydro-1H-cyclohexane[1,2,3-de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl]amino}carbamate

[1502]

[1503] Step 22a. Add 1-methylimidazolium (1.359 mL, 17.051 mmol) to a mixture of (2S)-2-({[(9H-fluoren-9-ylmethyl)oxy]carbonyl}amino)-5-[(2-methylprop-2-yl)oxy]-5-oxovalerate (3.97 g, 9.337 mmol) and (4-aminophenyl)methanol (1 g, 8.120 mmol) in DMF (18.34 mL). Add TCFH (2.73 g, 9.743 mmol) to the mixture. Stir the mixture at 23 °C for 2 hours. Concentrate the mixture under reduced pressure and partition the residue between EA (100 mL) and water (50 mL). Dilute the aqueous layer with EA (30 mL) 2) Extraction was performed, and the combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid chromatography (160 g silica column, PE / EA, where EA ranged from 0-63%) to provide the desired product (3.2 g, white solid, yield: 74.3%). LC-MS: (ESI) m / z (M+Na), 553.3.

[1504] Step 22b. At 0°C, 4-nitrophenyl chloroformate (8.55 g, 42.403 mmol) was added to a mixture of the compound from step 22a (1.5 g, 2.827 mmol) and pyridine (1.137 mL, 14.134 mmol) in THF (50 mL) and DMF (25 mL). The mixture was stirred at 20°C for 3 hours. The mixture was concentrated under reduced pressure and the residue was partitioned between EA (100 mL) and water (50 mL). The aqueous layer was separated with EA (20 mL). 2) Extraction was performed, and the combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid chromatography (80 g silica column, PE / EA, where EA ranged from 0-55%) to provide the title product (1.46 g, colorless oil, yield: 74.2%). LC-MS: (ESI) m / z (M+Na), 718.4.

[1505] Step 22c. Add the compound from step 22b (225 mg, 0.323 mmol) to a mixture of (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,10,12,13,15-octahydrocyclohexane[1,2,3-de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-10,13-dione methanesulfonic acid (257.86 mg, 0.485 mmol), DIPEA (0.160 mL, 0.970 mmol), and DMF (6 mL). Stir the mixture at 20 °C for 12 hours. The solution was filtered and the filtrate was purified by passing it through a reversed-phase column (80 g C18 gel column, H2O / CH3CN, where CH3CN ranged from 0-84%) to provide the desired product (140 mg, yellow solid, yield: 43.6%). LC-MS: (ESI) m / z (M+H), 992.4.

[1506] Step 22d. Diethylamine (154.8 mg, 2.11 mmol) in DMF (5 mL) was added to a solution of the compound (140 mg, 0.14 mmol) from step 22c at 0 °C for 2 hours. The solution was filtered and concentrated to obtain the desired compound (0.12 g, white solid, quantitative yield), which could be used in the next step without purification. LC-MS: (ESI) m / z (M+H), 770.4.

[1507] Step 22. Add the compound from step 5h (145 mg, 0.32 mmol) to a mixture of the compound from step 22d (124 mg, 0.16 mmol) and DIPEA (62.3 mg, 0.48 mmol) in DMF (3 mL). Stir the mixture at 20 °C for 2 h. Remove the solvent under vacuum and purify the residue by preparative HPLC (C18, 20%–60% acetonitrile in H2O (containing 0.1% TFA)) to provide the title compound LP22 (278 mg, 15.7% yield) as a yellow solid. LC-MS: (ESI) m / z (M+H), 1108.6. 1H NMR (400 MHz, DMSO) δ 9.80 (s, 1H), 8.20 -8.18 (m, 1H), 8.00 - 7.99 (m, 1H), 7.96 - 7.86 (m, 2H), 7.75 - 7.72 (m, 1H),7.60 - 7.56 (m, 2H), 7.33 - 7.30 (m, 2H), 7.24 (s, 1H), 6.93 (s, 2H), 6.45(s, 1H), 5.38 (s, 2H), 5.25 - 5.23 (m, 3H), 5.01 (s, 2H), 4.39 - 4.31 (m,1H), 4.20 - 4.14 (m, 1H), 3.70 - 3.67 (m, 2H), 3.53 - 3.51 (m, 3H), 2.31 (s,3H), 2.10 - 2.04 (m, 6H), 1.85 - 1.72 (m, 6H), 1.43 - 1.40 (m, 6H), 1.30 (s,9H), 1.14 - 1.10 (m, 2H), 0.81 (t, J = 7.2 Hz, 3H).

[1508] Step 21. Add TFA (0.04 mL, 0.54 mmol) to a mixture of the compound from Step 22 (i.e., Example 22, 20 mg, 0.018 mmol) in DCM (0.4 mL). Stir the mixture at 0 °C for 4 hours. Remove the solvent under vacuum and purify the residue by preparative HPLC (C18, 30%–53% acetonitrile in H2O (containing 0.1% TFA)) to provide the title compound LP21 (i.e., Example 21, 6.4 mg, 34.2% yield) as a yellow solid. LC-MS: (ESI) m / z (M+H), 1052.4. 1H NMR (400 MHz, DMSO) δ 12.13 (s, 1H), 9.89 (s, 1H), 8.30 - 8.26 (m,1H), 8.09 - 8.07 (m, 1H), 8.05 - 7.99 (m, 2H), 7.81 - 7.79 (m, 1H), 7.65 -7.63 (m, 2H), 7.40 - 7.38 (m, 2H), 7.32 (s, 1H), 6.99 (s, 2H), 6.53 (s, 1H), 5.45 (s, 2H), 5.30 (s, 3H), 5.09 (s, 2H), 4.45 - 4.39 (m, 1H), 4.25 - 4.22(m, 1H), 3.78 - 3.75 (m, 2H), 3.62 - 3.58 (m, 2H), 3.14 (s, 1H), 2.38 (s,3H), 2.32 - 2.11 (m, 8H), 2.10 - 1.87 (m, 6H), 1.50 - 1.47 (m, 4H), 1.21 –1.15 (m, 2H), 0.88 (t, J = 7.2 Hz, 3H).

[1509] Example 23: Preparation of LP23 (MC-SGC(Me)-PABC-Ecinotecan)

[1510]

[1511] 4-((S)-2-(2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)hexamido)-3-hydroxypropamido)acetamyl)-5-(3-methylurea)pentamido)benzyl((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl)carbamate

[1512]

[1513] Step 23a. At 0 °C, TEA (3.530 mL, 25.394 mmol) and 1-(chlorooxyalkyl)-N-methylformamide (1.85 g, 16.930 mmol) were slowly added to a solution of (2S)-5-amino-2-({[(9H-fluorene-9-ylmethyl)oxy]carbonyl}amino)valerate (3 g, 8.465 mmol) in DCM (50 mL). The resulting mixture was stirred at 20 °C for about 3 h. It was concentrated under reduced pressure and the residue was partitioned between EA (50 mL) and water (20 mL). The aqueous layer was extracted with EA (20 mL × 2), and the combined organic matter was dried (Na2SO4), filtered, and concentrated. The residue was purified by chromatography (0-10% DCM in silica, MeOH) to provide the desired compound (2.2 g, white solid, 63% yield). LC-MS: (ESI) m / z (M+Na), 412.2.

[1514] Step 23b. At 0 °C, (4-aminophenyl)methanol (0.66 g, 5.347 mmol), 1-methyl-1H-imidazole (0.92 g, 11.228 mmol), and TCFH (1.80 g, 6.416 mmol) were slowly added to a solution of the compound from step 23a (2.2 g, 5.347 mmol) in DMF (20 mL). The resulting mixture was stirred at 20 °C for about 16 h. It was concentrated under reduced pressure and the residue was partitioned between EA (50 mL) and water (30 mL). The aqueous layer was extracted with EA (30 mL × 2), and the combined organic matter was dried (Na2SO4), filtered, and concentrated. The residue was purified by chromatography (silica, 0-10% MeOH in DCM) to provide the desired compound (450 mg, white solid, 16% yield). LC-MS: (ESI) m / z (M+H), 517.2.

[1515] Step 23c. At 0 °C, DIPEA (0.288 mL, 1.742 mmol) and 4-nitrophenyl[(4-nitrophenyl)oxy]carbamate (1059.97 mg, 3.484 mmol) were slowly added to a solution of the compound from Step 23b (300 mg, 0.581 mmol) in DMF (10 mL). The resulting mixture was stirred at 20 °C for about 16 h. It was concentrated under reduced pressure and the residue was partitioned between EA (20 mL) and water (10 mL). The aqueous layer was extracted with EA (10 mL × 2), and the combined organic matter was dried (Na2SO4), filtered, and concentrated. The residue was purified by chromatography (silica, 0-10% MeOH in DCM) to provide the desired compound (170 mg, yellow solid, 43% yield). LC-MS: (ESI) m / z (M+H), 680.0.

[1516] Step 23d. At 0 °C, DIPEA (0.048 mL, 0.293 mmol) and (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,10,12,13,15-octahydrocyclohexane[1,2,3-de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-10,13-dione (127.76 mg, 0.293 mmol) were slowly added to a solution of the compound from step 23c (200 mg, 0.293 mmol) in DMF (5 mL). The resulting mixture was stirred at 20 °C for about 1 h. It was concentrated under reduced pressure and the residue was purified by RP-FC (5%-95% MeCN in C18, H2O) to provide the desired compound (250 mg, yellow solid, 87% yield). LC-MS: (ESI) m / z (M+H), 978.4.

[1517] Step 23e. Diethylamine (0.063 mL, 0.613 mmol) was slowly added to a solution of the compound from step 23d (200 mg, 0.204 mmol) in DMF (5 mL) at 0 °C. The resulting mixture was stirred at 20 °C for about 2 h. It was concentrated under reduced pressure, and the mixture was concentrated to obtain the desired compound, which could be used in the next step without purification (154 mg, yellow solid, quantitative yield). LC-MS: (ESI) m / z (M+H), 756.2.

[1518] Step 23f. At 0 °C, DIPEA (0.098 mL, 0.595 mmol) and the compound from step 3h (89.79 mg, 0.198 mmol) were slowly added to a solution of the compound from step 23e (150 mg, 0.198 mmol) in DMF (5 mL). The resulting mixture was stirred at 25 °C for about 1 h. It was concentrated under reduced pressure and the residue was purified by preparative HPLC (C18, 20%–60% acetonitrile in H2O (containing 0.1% formic acid)) to provide the title compound (28.3 mg, yellow solid, 13% yield). LC-MS: (ESI) m / z (M+H), 1094.4. 1 H NMR (400 MHz, DMSO) δ 9.93 (s, 1H),8.21 (t, J = 5.6 Hz, 1H), 8.05 (d, J = 8.8 Hz, 1H), 7.99 (d, J = 7.6 Hz, 1H),7.93 (d, J = 7.2 Hz, 1H), 7.77 (d, J = 10.8 Hz, 1H), 7.62 (d, J = 8.4 Hz,2H), 7.37 (d, J = 8.4 Hz, 2H), 7.31 (s, 1H), 6.98 (s, 2H), 6.52 (s, 1H),5.95-5.91 (m, 1H), 5.70-5.60 (m, 1H), 5.45 (s, 2H), 5.28-5.27 (m, 3H), 5.08(s, 2H), 4.41-4.36 (m, 1H), 4.25-4.21 (m, 1H), 3.75-3.74 (m, 2H), 3.58-3.56(m, 2H), 3.28-3.19 (m, 2H), 3.09-2.89 (m, 4H), 2.37 (s, 4H), 2.22-2.11 (m,5H), 1.92-1.83 (m, 2H), 1.74-1.65 (m, 1H), 1.61-1.57 (m, 1H), 1.56-1.48 (m,6H), 1.37-1.31 (m, 1H), 1.22-1.15 (m, 3H), 0.88 (t, J = 7.2 Hz, 3H).

[1519] Example 24: Preparation of LP24 (MC-SGC(PEG)-PABC-Ecinotecan)

[1520]

[1521] 4-((S)-2-(2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)hexamido)-3-hydroxypropamido)acetamyl)-5-(3-(2-hydroxyethyl)ureo)pentamido)benzyl((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-1,2,3,9,10,12,13,15-octahydrobenzo[de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl)carbamate

[1522]

[1523] Step 24a. At 20°C, TBDPSCl (14.18 g, 51.57 mmol) was added to a solution of 2-aminoethanol (3 g, 49.16 mmol) and 1H-imidazole (6.69 g, 98.23 mmol) in DCM (50 mL). After stirring for 18 hours, the mixture was concentrated under reduced pressure and the residue was partitioned between DCM (100 mL) and water (50 mL). The aqueous layer was then separated with DCM (50 mL). 2) Extraction was performed, and the combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid chromatography (80 g silica gel column, DCM / MeOH, where MeOH ranged from 0–5%) to provide the desired product (7.5 g, colorless oil, yield: 53.57%). LC-MS: (ESI) m / z (M+H), 300.2.

[1524] Step 24b. At 25 °C, the compound from step 24a (3.08 g, 12.02 mmol) was added to a solution of 2-((tert-butyldiphenylsilyl)oxy)ethylamine (3 g, 10.01 mmol) in DCM (30 mL). After stirring for 3 hours, the solution was concentrated and the residue was purified by rapid chromatography (80 g silica gel column, DCM / MeOH, where MeOH ranged from 0–5%) to provide the desired product (3 g, colorless oil, yield: 67.98%). LC-MS: (ESI) m / z (M+Na), 463.2.

[1525] Step 24c. Add DIPEA (2.64 g, 20.43 mmol) to a mixture of the compound from Step 24b (3 g, 6.81 mmol) and (2S)-5-amino-2-({[(9H-fluorene-9-ylmethyl)oxy]carbonyl}amino)valeric acid (2.9 g, 8.17 mmol) in DMF (8 mL). Stir the mixture at 25 °C for 18 hours. Filter the solution and purify the filtrate by passing it through a reversed-phase column (80 g C18 gel column, H2O / CH3CN, where CH3CN ranges from 0–80%) to provide the desired product (0.36 g, white solid, yield: 7.78%). LC-MS: (ESI) m / z (M+H), 680.2.

[1526] Step 24d. At 0°C, 1-methylimidazole (80 mg, 0.97 mmol) was added to a mixture of the compound from step 24c (300 mg, 0.44 mmol), (4-aminophenyl)methanol (70 mg, 0.52 mmol), and TCFH (150 mg, 0.52 mmol) in DMF (5 mL). The resulting mixture was then stirred at 0°C for 2 hours, concentrated under reduced pressure, and the residue was partitioned between DCM (10 mL) and water (5 mL). The aqueous layer was then separated with DCM (5 mL). 2) Extraction was performed, and the combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid chromatography (10 g silica column, DCM / MeOH, where MeOH ranged from 0–5%) to provide the desired product (0.24 g, yellow solid, yield: 69.29%). LC-MS: (ESI) m / z (M+Na), 785.2.

[1527] Step 24e. At 0 °C, DIPEA (74.09 g, 0.57 mmol) was added to a solution of the compound from Step 24d (150 mg, 0.19 mmol), 4-nitrophenyl[(4-nitrophenyl)oxy]carbamate (116.25 g, 0.38 mmol), and 4-(dimethylamino)pyridine (11.67 mg, 8.96 mmol) in DMF (5 mL). After stirring at 25 °C for 18 hours, the solution was diluted with DCM (20 mL) and washed with water (10 mL), and the phases were separated. The aqueous layer was extracted four times with DCM (20 mL), and the combined organic phases were dried over sodium sulfate and the solvent was removed under reduced pressure. The residue was purified by rapid chromatography (10 g silica gel column, DCM / MeOH, where MeOH ranged from 0–5%) to provide the desired product (0.24 g, yellow solid, yield: 69.29%). LC-MS: (ESI) m / z (M+H), 951.4.

[1528] Step 24f. At 20°C, DIPEA (49 mg, 0.38 mmol) was added to a solution of the compound from step 24e (120 mg, 0.23 mmol), (1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,10,12,13,15-octahydrocyclohexane[1,2,3-de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-10,13-dione methanesulfonic acid (80 mg, 0.15 mmol) in DMF (10 mL). After stirring at 20°C for 4 hours, the solution was concentrated and the residue was purified by rapid chromatography (10 g silica gel column, DCM / MeOH, where MeOH ranged from 0–5%) to provide the desired product (0.11 g, yellow solid, yield: 69.87%). LC-MS: (ESI) m / z (1 / 2M+H), 624.0.

[1529] Step 24g. At 0°C, 1N HCl (0.08 ml, 0.08 mmol) was added to a solution of the compound from Step 24f (100 mg, 0.08 mmol), CH3COOH (0.08 ml, 1.39 mmol), and THF (5 mL). After stirring for 18 hours, the solution was filtered, and the filtrate was purified by preparative HPLC to provide the desired product (40 mg, yellow solid, yield: 49.46%). LC-MS: (ESI) m / z (M+H), 1009.4.

[1530] Step 24h. Diethylamine (43.5 mg, 0.59 mmol) was added to a solution of the compound from step 24g (40 mg, 0.04 mmol) in DMF (5 mL) at 0 °C for 2 hours. After stirring for 2 hours, the solution was filtered and concentrated to obtain the desired compound (50 mg, yellow oil, quantitative yield), which could be used in the next step without purification. LC-MS: (ESI) m / z (M+H), 786.4.

[1531] Step 24i. At 25 °C, DIPEA (24.7 mg, 0.19 mmol) and the compound from step 3 h (57.57 mg, 0.13 mmol) were added to a solution of the compound from step 24 h (50 mg, 0.064 mmol) in DMF (3 mL). After stirring at 25 °C for 2 hours, the solvent was removed under vacuum and the residue was purified by preparative HPLC (C18, H2O (containing 0.1% TFA) in 20%–60% acetonitrile) to provide the title compound as a yellow solid (18.5 mg, 25.9% yield). LC-MS: (ESI) m / z (M+H), 1124.6. 1H NMR (400 MHz, DMSO) δ 9.93 (s, 1H), 8.21 (s,1H), 8.10 - 8.06 (m, 1H), 8.05 - 8.00 (m, 1H), 7.95 - 7.93 (m, 1H), 7.80 -7.78 (m, 1H), 7.65 - 7.62 (m, 2H), 7.41 - 7.37 (m, 2H), 7.31 (s, 1H), 7.00(s, 2H), 6.51 (s, 1H), 5.97 (s, 1H), 5.84 (s, 1H), 5.45 (s, 2H), 5.30 - 5.29(m, 3H), 5.08 (s, 2H), 4.40 - 4.38 (m, 1H), 4.25 - 4.21 (m, 1H), 3.80 - 3.75(m, 2H), 3.59 - 3.56 (m, 2H), 3.25 - 3.14 (m, 4H), 3.03 - 2.95 (m, 0.88 (t, J = 7.2 Hz, 3H).

[1532] Example 25: Preparation of LP25 (MC-TGC-PABC-Ecinotecan)

[1533]

[1534] 4-((S)-2-(2-((2S,3S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrolo-1-yl)hexamido)-3-hydroxybutamido)acetamido)-5-ureidopentamido)benzyl((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]inzizo[1,2-b]quinoline-1-yl)carbamate

[1535]

[1536] Step 25a. At 0 °C, HATU (2.87 g, 7.553 mmol), DIPEA (2.488 mL, 15.105 mmol), and glycine tert-butyl ester (660 mg, 5.035 mmol) were slowly added to a solution of N-(((9H-fluorene-9-yl)methoxy)carbonyl)-O-(tert-butyl)-L-allothreonine (2.0 g, 5.035 mmol) in DMF (20 mL). The resulting mixture was stirred at 25 °C for about 16 h. It was concentrated under reduced pressure and the residue was partitioned between EA (10 mL) and water (5 mL). The aqueous layer was extracted with EA (10 mL × 2), and the combined organic matter was dried (Na2SO4), filtered, and concentrated. The residue was purified by chromatography (silica, 0-10% MeOH in DCM) to provide the desired compound (1.8 g, white solid, 70% yield). LC-MS: (ESI) m / z (M+Na), 533.4.

[1537] Step 25b. At 0 °C, TFA (3 mL) was slowly added to a solution of the compound from step 25a (700 mg, 1.372 mmol) in DCM (15 mL). The resulting mixture was stirred at 25 °C for approximately 16 h. It was concentrated under reduced pressure and the residue was purified by RP-FC (C18, 5%–95% MeCN in H2O) to provide the desired compound (300 mg, white solid, 54% yield). LC-MS: (ESI) m / z (M+Na), 399.0.

[1538] Step 25c. At 0 °C, HATU (115.34 mg, 0.303 mmol), DIPEA (0.100 mL, 0.607 mmol), and the compound from step 25b (96.68 mg, 0.243 mmol) were slowly added to a solution of the compound from step 19d (150 mg, 0.202 mmol) in DMF (5 mL). The resulting mixture was stirred at 25 °C for about 8 h. It was concentrated under reduced pressure and the residue was partitioned between EA (30 mL) and water (15 mL). The aqueous layer was extracted with EA (20 mL × 2), and the combined organic matter was dried (Na2SO4), filtered, and concentrated. The residue was purified by RP-FC (C18, 5%–95% MeCN in H2O) to provide the desired compound (60 mg, white solid, 26% yield). LC-MS: (ESI) m / z (M+H), 1122.4.

[1539] Step 25d. Diethylamine (0.017 mL, 0.160 mmol) was slowly added to a solution of the compound from step 25c (60 mg, 0.053 mmol) in DMF (0.5 mL) at 0 °C. The resulting mixture was stirred at 0 °C for about 2 h. It was concentrated under reduced pressure and the residue was purified by RP-FC (C18, 5%–95% MeCN in H2O) to provide the desired compound (35 mg, white solid, 72% yield). LC-MS: (ESI) m / z (M+H), 900.2.

[1540] Step 25e. At 0 °C, DIPEA (0.019 mL, 0.117 mmol) and 1-{6-[(2,5-dioxotetrahydro-1H-pyrrolo-1-yl)oxy]-6-oxohexyl}pyrrolo-2,5-dione (17.98 mg, 0.058 mmol) were slowly added to a solution of the compound from Step 25d (35 mg, 0.039 mmol) in DMF (1 mL). The resulting mixture was stirred at 25 °C for about 1 h. It was concentrated under reduced pressure and the residue was purified by preparative HPLC (C18, H2O (containing 0.1% formic acid) in 20%–60% acetonitrile) to prov...

Claims

1. A compound having formula (X), or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, diastereomer, or mixture thereof: SD (X) in, S is a linker, and the linker contains a peptide-cleavable unit that contains a tripeptide having the sequence -P3-P2-P1-. P3 is selected from serine, tyrosine, or their analogues; P2 is selected from glycine, serine, or their analogues; P1 is selected from citrulline, arginine, glutamic acid or their analogues; D represents the drug component.

2. The compound of claim 1, wherein, The connector S further includes a cuttable connector or a non-cuttable connector; Alternatively, the cleavable linker may comprise an acid-instable linker, a hydrophilic linker, a protease-sensitive linker, a light-instable linker, a hydrazone linker, a dimethyl linker, or a disulfide-containing linker.

3. The compound according to claim 1 or 2, wherein, The compound has the structure of formula (I), or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, diastereomer, or mixture thereof: LPYD (I) L is represented by L1-L2-X-; L1 is the ligand covalently bound portion. -L2-X- is an extended subunit; Y is the spacer subunit; P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y; P3 is selected from serine, tyrosine, or their analogues; P2 is selected from glycine, serine, or their analogues; P1 is selected from citrulline, arginine, glutamic acid, or their analogues. D represents the drug component.

4. The compound according to any one of claims 1-3, in, P3 is , Rc1, Rc2, and Rc3 are independently selected from H, OH, CN, NH2, halogens, O-PG, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 3-10 Cycloalkyl, 3- to 10-membered heterocyclic, 5- to 10-membered heteroaryl and C 6-10 Aryl; Or Rc2 and Rc3 together with the carbon atoms to which they are attached form C 3-10 Cycloalkylene or 3- to 10-membered heterocyclic cycloidene groups; c is 0, 1, 2, 3 or 4; P3 may optionally be further substituted with 1, 2, 3, 4 or 5 Rc groups; Rc is selected from H, OH, CN, NH2, halogens, PG, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl groups and C 1-6 The haloalkoxy group can be alternatively selected from H, OH, CN, NH2, halogen, PG, C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups; P2 is Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl, C 1-6 Halogenated alkyl groups and -(CH2) 1-6 -OH, or Rb1 and Rb2 together with the carbon atoms to which they are attached, form C 3-10 Cycloalkylene or 3- to 10-membered heterocyclic cycloidene groups; P2 may optionally be further substituted with 1, 2, 3, 4 or 5 Rb groups; Rb is selected from H, halogens, PG, and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups; P1 is selected from ; La is -C 1-8 alkylene-, where -C 1-8 The 1, 2, 3 or 4 non-adjacent carbon atoms in the alkylene group can be optionally replaced by O or S; Xa is a bond or -NR-; R is H or -C 1-4 alkyl; R1 is selected from H, Boc, and C. 1-6 Alkyl groups, -OR', NR'R'', -C(O)R', -C(O)OR', -C(NH)NR'R'', and -C(O)NR'R'', preferably selected from H, Boc, C 1-6 Alkyl, -OR', -C(O)R', -C(O)OR', -C(NH)NR'R'' and -C(O)NR'R''; R' and R'' are independently selected from H, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, -C 1-6 Alkylene-CN, -C 1-6 Alkyl-NH2 and -C 1-6 alkylene-OH; P1 may optionally be further substituted with 1, 2, 3, 4 or 5 Ra groups; Ra is selected from H, halogen, PG, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; " "Represents a chiral center, which is selected from the (S) or (R) configuration; PG is selected from: trimethylsilyl group (TMS), triethylsilyl group (TES), dimethylisopropylsilyl group (DMIPS), diethylisopropylsilyl group (DEIPS), tert-butyldimethylsilyl group (TBDMS), tert-butyldiphenylsilyl group (TBDPS), triisopropylsilyl group (TIPS), acetyl group (Ac), chloroacetyl group, dichloroacetyl group, trichloroacetyl group, trifluoroacetyl group (TFA), benzoyl group, p-methoxybenzoyl group, 9-fluorenylmethoxycarbonyl group (Fmoc), allyloxycarbonyl group (Alloc), 2,2 2-Trichloroethoxycarbonyl (Troc), carboxybenzyl (Cbz), tert-butoxycarbonyl (Boc), benzyl (Bn), p-methoxybenzyl (PMB), allyl, triphenylmethyl (Tr), bis-p-methoxytriphenylmethyl (DMTr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), p-methoxybenzyloxymethyl (PMBM), -C(O)CH2CH2C(O)OH or 4,4'-dimethoxytriphenylmethyl, alternatively, Alternatively, P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y; Among them, P3 is , Rc1 is selected from H, OH, O-PG, and C. 1-4 The alkoxy groups, Rc2 and Rc3, are independently selected from H, halogens, and C. 1-4 Alkyl, 5- to 10-membered heteroaryl (e.g., 5- to 6-membered heteroaryl) and C 6-10 Aryl groups (such as phenyl), or Rc2 and Rc3, together with the carbon atoms to which they are attached, form C 3-5 Cycloalkylene; Rc1, Rc2 and Rc3 can be independently and optionally replaced by 1, 2 or 3 Rc; Rc is selected from H, OH, C 1-4 Alkyl, C 1-4 Alkoxy and C 1-4 Haloalkyl groups, which may be selected from H, OH, C. 1-4 Alkyl and C 1-4 Halogenated alkyl groups; c is 0, 1, or 2; P2 is Rb1 and Rb2 are independently selected from H and C. 1-4 Alkyl groups and -(CH2) 1-4 -OH, or Rb1 and Rb2 together with the carbon atoms to which they are attached, form C 3-5 Cycloalkylene; P1 is selected from ; La is -(CH2) 1-6 -, where -(CH2) 1-6 The 1, 2, or 3 non-adjacent carbon atoms in - can be optionally replaced by O; alternatively, La is selected from -CH2-, -CH2CH2-, CH2CH2CH2-, CH2CH2CH2CH2-, and -CH2OCH2CH2-; Xa is a bond or -NR-; R is H or -C 1-4 alkyl; R1 is selected from H, Boc, NR'R'', and C. 1-4 Alkyl groups, -C(NH)NR'R'', -C(O)NR'R'', and -C(O)OR', preferably selected from H, Boc, and C 1-4 Alkyl groups, -C(NH)NR'R'', -C(O)NR'R'', and -C(O)OR'; R' and R'' are independently selected from H and -C. 1-4 Alkyl and -C 1-4 alkylene-OH; Alternatively, R1 can be selected from Boc, NH2, and C. 1-4 Alkyl groups, -C(NH)NH2, -C(O)NH2, -C(O)NH-C 1-4 Alkyl group, -C(O)NH-C 1-4 Alkylenes -OH, -C(O)OH and -C(O)OC 1-4 alkyl; " "Represents a chiral center, which is selected from the (S) or (R) configuration; Alternatively, P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y; P3 is selected from: , , , , , , , , , and Alternatively, it can be selected from: , , , , , , and ; P2 is selected from: , , and ; P1 is selected from: , , , , , , , , , , , , and .

5. The compound according to claim 3 or 4, wherein, L is represented by L1-L2-X-; L1 is the ligand covalently bound part. Alternatively, L1 can form a covalent bond with the functional group of an amino acid, and even more alternatively, L1 can form an amide bond or a thioether bond with the functional group of an amino acid. -L2-X- is an extended subunit; L2 is selected from C 1-20 Alkylene, C 2-20 imide and C 2-20 Idemynyl group, in which C 1-20 The 1, 2, 3, 4, 5, 6, 7 or 8 non-adjacent carbon atoms in the alkylene group may optionally be replaced by O or S; X either does not exist or is -C(O)-; L can be arbitrarily divided by 1, 2, 3, 4 or 5 Rs. L Group substitution; R L Selected from H, D, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; Alternatively, L is represented by L1-L2-X-; L1 is a ligand covalently bonded moiety capable of forming amide or thioether bonds with the functional groups of amino acids, and can alternatively be selected from... and ; -L2-X- is an extended subunit; L2 is selected from C 1-10 Alkylene, -C 0-4 Alkylene-(C 1-4 alkylene-O) 1-10 -C 0-4 alkylene-, -C 0-4 Alkylene-(OC) 1-4 Alkylene) 1-10 -C 0-4 Alkylene-, C 2-10 imide and C 2-10 Ethyne group; X either does not exist or is -C(O)-; L can be arbitrarily divided by 1, 2, 3, 4 or 5 Rs. L Group substitution; R L Selected from H, D, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; Alternatively, L is represented by L1-L2-X-; L1 is selected from and ; L2 is selected from C 1-10 Alkylene, -(CH2) 0-4 -(CH2CH2-O) 1-10 -(CH2) 0-4 -、-(CH2) 0-4 -(O-CH2CH2) 1-10 -(CH2) 0-4 -、C 2-10 imide and C 2-10 Ethyne group; X either does not exist or is -C(O)-; L can be optionally divided by 1, 2 or 3 Rs L Group substitution; R L Selected from H, D, C 1-4 Alkyl and C 1-4 Halogenated alkyl groups; Alternatively, L is selected from , and ; m and n are independently selected from 1, 2, 3, 4, 5, 6, 7 or 8; Alternatively, m can be selected from 4, 5, 6, 7 or 8, and can be 4, 5 or 8; Alternatively, n can be selected from 2, 3, or 4, and 3 is an alternative.

6. The compound according to any one of claims 3-5, wherein, Y is selected from: and ; R Y1 Selected from H, halogens, OH, CN, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl groups and C 1-6 Halogenated alkoxy groups; R Y2 Selected from H, halogens, OH, CN, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl groups and C 1-6 Halogenated alkoxy groups; p is 0, 1, or 2; q is 0, 1, 2, 3, or 4; Alternatively, Y is selected from: and ; R Y1 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; R Y2 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; p is 0, 1, or 2; q is 0, 1, or 2; Alternatively, Y is selected from: (AM) and (PABC).

7. The compound according to any one of claims 1-6, wherein, Part D of this drug is selected from the following groups: microtubule inhibitors and microtubule polymerization inhibitors, antibiotics, DNA synthesis inhibitors, topoisomerase I inhibitors, immunomodulators, and chelating ligands; Alternatively, The microtubule inhibitors and microtubule polymerization inhibitors are selected from the group consisting of: olistatins (e.g., MMAE, MMAF, and MMAD), maytansines (e.g., maytansine, maytanol, DM1, DM2, DM3, and DM4), tubulosyntheticin, candidin, eribulin, and rhizobium. This antibiotic was selected from the group consisting of: chalcone, doxorubicin, and anthracyclines; The DNA synthesis inhibitor was selected from the group consisting of: betamethasone, PBD (pyrrolobenzodiazepine), and IGN (indolinobenzodiazepine). The topoisomerase I inhibitor is selected from the group consisting of: camptothecin and camptothecin analogues (e.g., DXd, SN38 and ixotecan). The immunomodulator was selected from the following group: TLR7 agonists, TLR8 agonists, STING agonists, and RIG-I agonists; Alternatively, D is selected from: , and .

8. The compound according to any one of claims 3-7, wherein, L is represented by L1-L2-X-; L1 is a ligand covalently bonded moiety capable of forming amide or thioether bonds with the functional groups of amino acids, and can alternatively be selected from... and ; -L2-X- is an extended subunit; L2 is selected from C 1-10 Alkylene, -(C 1-4 alkylene-O) 1-10 -、-(OC 1-4 Alkylene) 1-10 -、-C 2-10 imide and C 2-10 Ethyne group; X either does not exist or is -C(O)-; L can be arbitrarily divided by 1, 2, 3, 4 or 5 Rs. L Group substitution; R L Selected from H, D, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; Y is selected from: and ; R Y1 Selected from H, halogens, OH, CN, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl groups and C 1-6 Halogenated alkoxy groups; R Y2 Selected from H, halogens, OH, CN, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl groups and C 1-6 Halogenated alkoxy groups; p is 0, 1, or 2; q is 0, 1, 2, 3, or 4; Part D of this drug is selected from the following groups: microtubule inhibitors and microtubule polymerization inhibitors, antibiotics, DNA synthesis inhibitors, topoisomerase I inhibitors, immunomodulators, and chelating ligands; P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y; Among them, P3 is , Rc1, Rc2, and Rc3 are independently selected from H, OH, CN, NH2, halogens, O-PG, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 3-10 Cycloalkyl, 3- to 10-membered heterocyclic, 5- to 10-membered heteroaryl and C 6-10 Aryl; Or Rc2 and Rc3 together with the carbon atoms to which they are attached form C 3-10 Cycloalkylene or 3- to 10-membered heterocyclic cycloidene groups; c is 0, 1, 2, 3 or 4; P3 may optionally be further substituted with 1, 2, 3, 4 or 5 Rc groups; Rc is selected from H, OH, CN, NH2, halogens, PG, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl groups and C 1-6 The haloalkoxy group can be alternatively selected from H, OH, CN, NH2, halogen, PG, C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups; P2 is Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl, C 1-6 Halogenated alkyl groups and -(CH2) 1-6 -OH, or Rb1 and Rb2 together with the carbon atoms to which they are attached, form C 3-10 Cycloalkylene or 3- to 10-membered heterocyclic cycloidene groups; P2 may optionally be further substituted with 1, 2, 3, 4 or 5 Rb groups; Rb is selected from H, halogens, PG, and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups; P1 is selected from ; La is -C 1-8 alkylene-, where -C 1-8 The 1, 2, 3 or 4 non-adjacent carbon atoms in the alkylene group can be optionally replaced by O or S; Xa is a bond or -NR-; R is H or -C 1-4 alkyl; R1 is selected from H, Boc, and C. 1-6 Alkyl groups, -OR', NR'R'', -C(O)R', -C(O)OR', -C(NH)NR'R'', and -C(O)NR'R'', preferably selected from H, Boc, C 1-6 Alkyl, -OR', -C(O)R', -C(O)OR', -C(NH)NR'R'' and -C(O)NR'R''; R' and R'' are independently selected from H, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, -C 1-6 Alkylene-CN, -C 1-6 Alkyl-NH2 and -C 1-6 alkylene-OH; P1 may optionally be further substituted with 1, 2, 3, 4 or 5 Ra groups; Ra is selected from H, halogen, PG, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; " "Represents a chiral center, which is selected from the (S) or (R) configuration; PG is selected from: trimethylsilyl group (TMS), triethylsilyl group (TES), dimethylisopropylsilyl group (DMIPS), diethylisopropylsilyl group (DEIPS), tert-butyldimethylsilyl group (TBDMS), tert-butyldiphenylsilyl group (TBDPS), triisopropylsilyl group (TIPS), acetyl group (Ac), chloroacetyl group, dichloroacetyl group, trichloroacetyl group, trifluoroacetyl group (TFA), benzoyl group, p-methoxybenzoyl group, 9-fluorenylmethoxycarbonyl group (Fmoc), allyloxycarbonyl group (Alloc). 2,2,2-Trichloroethoxycarbonyl (Troc), carboxybenzyl (Cbz), tert-butoxycarbonyl (Boc), benzyl (Bn), p-methoxybenzyl (PMB), allyl, triphenylmethyl (Tr), bis-p-methoxytriphenylmethyl (DMTr), methoxymethyl (MOM), phenoxymethyl (BOM), 2,2,2-trichloroethoxymethyl, 2-methoxyethoxymethyl (MEM), methylthiomethyl (MTM), p-methoxybenzyloxymethyl (PMBM), -C(O)CH2CH2C(O)OH or 4,4'-dimethoxytriphenylmethyl.

9. The compound according to any one of claims 3-8, wherein, L is represented by L1-L2-X-; L1 is selected from and ; L2 is selected from C 1-10 Alkylene, -(CH2) 0-4 -(CH2CH2-O) 1-10 -(CH2) 0-4 -、-(CH2) 0-4 -(O-CH2CH2) 1-10 -(CH2) 0-4 -、C 2-10 imide and C 2-10 Ethyne group; X either does not exist or is -C(O)-; L can be optionally divided by 1, 2 or 3 Rs L Group substitution; R L Selected from H, D, C 1-4 Alkyl and C 1-4 Halogenated alkyl groups; Y is selected from: and ; R Y1 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; R Y2 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; Part D of this drug is selected from the following groups: microtubule inhibitors and microtubule polymerization inhibitors, antibiotics, DNA synthesis inhibitors, topoisomerase I inhibitors, immunomodulators, and chelating ligands; The microtubule inhibitors and microtubule polymerization inhibitors are selected from the group consisting of: olistatins (e.g., MMAE, MMAF, and MMAD), maytansine (e.g., DM1, DM2, DM3, and DM4), tubulosyntheticin, candidin, eribulin, and rhizobium. This antibiotic was selected from the group consisting of: chalcone, doxorubicin, and anthracyclines; The DNA synthesis inhibitor was selected from the group consisting of: betamethasone, PBD (pyrrolobenzodiazepine), and IGN (indolinobenzodiazepine). The topoisomerase I inhibitor is selected from the group consisting of camptothecin and camptothecin analogues (e.g., DXd, SN38, and ixotecan). The immunomodulator was selected from the following group: TLR7 agonists, TLR8 agonists, STING agonists, and RIG-I agonists; P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y; Among them, P3 is , Rc1 is selected from OH, O-PG, and C. 1-4 The alkoxy groups, Rc2 and Rc3, are independently selected from H, halogens, and C. 1-4 Alkyl, 5- to 10-membered heteroaryl (e.g., 5- to 6-membered heteroaryl) and C 6-10 Aryl groups (such as phenyl), or Rc2 and Rc3, together with the carbon atoms to which they are attached, form C 3-5 Cycloalkylene; Rc1, Rc2 and Rc3 can be independently and optionally replaced by 1, 2 or 3 Rc; Rc is selected from H, OH, C 1-4 Alkyl, C 1-4 Alkoxy and C 1-4 Haloalkyl groups, which may be selected from H, OH, C. 1-4 Alkyl and C 1-4 Halogenated alkyl groups; c is 0, 1, or 2; P2 is Rb1 and Rb2 are independently selected from H and C. 1-4 Alkyl groups and -(CH2) 1-4 -OH, or Rb1 and Rb2 together with the carbon atoms to which they are attached, form C 3-5 Cycloalkylene; P1 is selected from ; La is -(CH2) 1-6 -, where -(CH2) 1-6 The 1, 2, or 3 non-adjacent carbon atoms in - can be optionally replaced by O; alternatively, La is selected from -CH2-, -CH2CH2-, CH2CH2CH2-, CH2CH2CH2CH2-, and -CH2OCH2CH2-; Xa is a bond or -NR-; R is H or -C 1-4 alkyl; R1 is selected from H, Boc, NR'R'', and C. 1-4 Alkyl groups, -C(NH)NR'R'', -C(O)NR'R'', and -C(O)OR', preferably selected from H, Boc, and C 1-4 Alkyl groups, -C(NH)NR'R'', -C(O)NR'R'', and -C(O)OR'; R' and R'' are independently selected from H and -C. 1-4 Alkyl and -C 1-4 alkylene-OH; Alternatively, R1 can be selected from Boc, NH2, and C. 1-4 Alkyl groups, -C(NH)NH2, -C(O)NH2, -C(O)NH-C 1-4 Alkyl group, -C(O)NH-C 1-4 Alkylenes -OH, -C(O)OH and -C(O)OC 1-4 alkyl; " "Represents a chiral center, which is selected from the (S) or (R) configuration.

10. The compound according to any one of claims 3-9, wherein, L is selected from , and ; m and n are independently selected from 1, 2, 3, 4, 5, 6, 7 or 8; Alternatively, m can be selected from 4, 5, 6, 7 or 8, and can be 4, 5 or 8; Alternatively, n can be selected from 2, 3, or 4, and 3 is an alternative. Y is selected from: (AM) and (PABC); D is selected from: , and ; P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y; P3 is selected from: , , , , , , , , , and Alternatively, it can be selected from: , , , , , , and ; P2 is selected from: , , and ; P1 is selected from: , , , , , , , , , , , , and .

11. The compound according to any one of claims 1-10, wherein, L, Y and D are as defined in any one of claims 3 and 5-10; P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y; -P3-P2-P1- is , " "Represents a chiral center, which is selected from the (S) or (R) configuration; Rc1, Rc2, and Rc3 are independently selected from H, OH, CN, NH2, halogens, and C. 1-6 Alkyl, C 1-6 Alkoxy and C 1-6 Halogenated alkyl groups; c is 0, 1, 2, 3 or 4; P3 may optionally be further substituted with 1, 2, 3, 4 or 5 Rc groups; Rc is selected from H, OH, CN, NH2, halogens, PG, and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups; Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl, C 1-6 Halogenated alkyl groups and -(CH2) 1-6 -OH; P2 may optionally be further substituted with 1, 2, 3, 4 or 5 Rb groups; Rb is selected from H, halogens, PG, and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups; La is -C 1-8 alkylene-, where -C 1-8 The 1, 2, 3 or 4 non-adjacent carbon atoms in the alkylene group can be optionally replaced by O or S; Xa is a bond or -NR-; R is H or -C 1-4 alkyl; R1 is selected from Boc, C 1-6 Alkyl groups, -C(O)R', -C(O)OR', -C(NH)NR'R'' and -C(O)NR'R''; R' and R'' are independently selected from H, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, -C 1-6 Alkylene-CN, -C 1-6 Alkyl-NH2 and -C 1-6 alkylene-OH; P1 may optionally be further substituted with 1, 2, 3, 4 or 5 Ra groups; Ra is selected from H, halogen, PG, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; alternative " "Represents a chiral center, which is selected from the (S) or (R) configuration; Rc1 is selected from H, OH, CN, NH2, halogens, and C. 1-6 Alkyl, C 1-6 Alkoxy and C 1-6 Halogenated alkyl groups; Rc2 and Rc3 are independently selected from H and C. 1-6 Alkyl, C 1-6 Alkoxy and C 1-6 Halogenated alkyl groups; c is 0, 1, 2, or 3; Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups; La is -C 1-6 alkylene-; Xa is -NR-; R is H or -C 1-4 alkyl; R1 is selected from -C(O)R', -C(O)OR', -C(NH)NR'R'' and -C(O)NR'R''; R' and R'' are independently selected from H, halogen, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; Alternatively, " "Represents a chiral center, which is selected from the (S) or (R) configuration; Rc1 is selected from H, OH, CN, NH2 and halogens, preferably OH; Rc2 and Rc3 are independently selected from H and C. 1-6 Alkyl groups, preferably H; c is 0, 1, or 2, preferably 0; Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl groups, preferably H; La is -C 1-6 alkylene-; Xa is -NH-; R1 is selected from -C(NH)NR'R'' and -C(O)NR'R'', preferably -C(NH)NR'R''; R' and R'' are independently selected from H and C. 1-6 Alkyl groups, preferably H; Alternatively, -P3-P2-P1- is (SGR).

12. The compound according to any one of claims 1-10, wherein, L, Y and D are as defined in any one of claims 3 and 5-10; P is a peptide cleavable unit, wherein the peptide cleavable unit contains a tripeptide having the sequence -P3-P2-P1-, P3 is linked to X, and P1 is linked to Y; -P3-P2-P1- is , " "Represents a chiral center, which is selected from the (S) or (R) configuration; Rc1, Rc2, and Rc3 are independently selected from H, OH, CN, NH2, halogens, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl groups, 5- to 10-membered heteroaryl groups and C 6-10 Aryl; c is 0, 1, 2, 3 or 4; P3 may optionally be further substituted with 1, 2, 3, 4 or 5 Rc groups; Rc is selected from H, OH, CN, NH2, halogens, PG, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl groups and C 1-6 Halogenated alkoxy groups; Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl, C 1-6 Halogenated alkyl groups and -(CH2) 1-6 -OH; P2 may optionally be further substituted with 1, 2, 3, 4 or 5 Rb groups; Rb is selected from H, halogens, PG, and C. 1-6 Alkyl and C 1-6 Halogenated alkyl groups; La is -C 1-8 alkylene-, where -C 1-8 The 1, 2, 3 or 4 non-adjacent carbon atoms in the alkylene group can be optionally replaced by O or S; Xa is a bond or -NR-; R is H or -C 1-4 alkyl; R1 is selected from H, Boc, and C. 1-6 Alkyl groups, -C(O)R', -C(O)OR', -C(NH)NR'R'' and -C(O)NR'R''; R' and R'' are independently selected from H, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, -C 1-6 Alkylene-CN, -C 1-6 Alkyl-NH2 and -C 1-6 alkylene-OH; P1 may optionally be further substituted with 1, 2, 3, 4 or 5 Ra groups; Ra is selected from H, halogen, PG, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; alternative " "Represents a chiral center, which is selected from the (S) or (R) configuration; Rc1 and Rc2 are independently selected from H, halogens, OH and C. 1-6 Alkoxy; Rc3 is selected from H, halogen, 5- to 10-membered heteroaryl (e.g., 5- to 6-membered heteroaryl), and C. 6-10 Aryl (e.g., phenyl); c is 0, 1, 2, or 3; Rc1, Rc2 and Rc3 are independently and optionally substituted by 1, 2 or 3 Rc groups; Rc is selected from H, OH, CN, NH2, halogens, PG, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl groups and C 1-6 Halogenated alkoxy groups; Rb1 and Rb2 are independently selected from H and C. 1-6 Alkyl groups and -(CH2) 1-6 -OH; La is -C 1-6 alkylene-; Xa is -NR-; R is H or -C 1-4 alkyl; R1 is selected from -C(O)R', -C(O)OR', -C(NH)NR'R'' and -C(O)NR'R''; R' and R'' are independently selected from H, halogen, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; Alternatively, " "Represents a chiral center, which is selected from the (S) or (R) configuration; Rc1 and Rc2 are independently selected from H, OH and C. 1-4 Alkyl group, preferably H; Rc3 is selected from H, halogen, 5- to 10-membered heteroaryl (e.g., 5- to 6-membered heteroaryl), and C. 6-10 Aryl (e.g., phenyl), preferably phenyl; c is 0, 1, or 2, preferably 0; Rc1, Rc2 and Rc3 are independently and optionally substituted by 1, 2 or 3 Rc groups; Rc is selected from H, OH, C 1-6 Alkyl and C 1-6 Alkoxy; Rb1 and Rb2 are independently selected from H and -(CH2). 1-4 -OH, preferably H or -CH2-OH; La is -C 1-6 alkylene-; Xa is -NH-; R1 is selected from -C(NH)NR'R'' and -C(O)NR'R'', preferably -C(NH)NR'R''; R' and R'' are independently selected from H and C. 1-6 Alkyl groups, preferably H; Alternatively, -P3-P2-P1- is (YSR).

13. The compound according to any one of claims 1-12, wherein, This compound was selected from: ； ； ； ； ； ； ； ； as well as ， Wherein, m, n, P3, P2 and P1 are as defined in any one of claims 1, 3-5 and 11-12; Alternatively, m is selected from 4, 5, 6, 7 or 8, and can be 4, 5 or 8; n is selected from 2, 3 or 4, and can be 3; Alternatively, the compound may be selected from: MC-VGR-DXD: 、 MC-SGR-DXD: 、 MC-VGR-PABC-MMAE: 、 MC-SGR-PABC-MMAE: 、 MC-SGC-DXD: 、 MC-SAC-DXD: 、 MsP-SGC-DXD: 、 MC-S(tBu)GC-DXD: 、 MC-SGOrn(Boc)-DXD: 、 MC-SGC-PABC-Eciticon: 、 MC-SGO(DEt)-PABC-Ecitidine: 、 MC-SGE-PABC-Ecitidine: 、 MC-SGE(tBu)-PABC-Eciticon: 、 MC-SGC(Me)-PABC-Eciticon: 、 MC-SGC(Peg)-PABC-Eciticon: 、 MC-TGC-PABC-Ecitidine: 、 MC-SGhC(O)-PABC-Ecitidine: 、 MC-YSR-PABC-Eciticon: 、 MC-SGE-DXD: 、 MC-SGC(Peg)-DXD: 、 MC-TSR-DXD: 、 MC-YSR-DXD: 、 MC-SGO-DXD: 、 MC-SGK-DXD: 、 Mal-PEG4-VGR-DXD: 、 Mal-PEG4-SGR-DXD: 、 MC-SGhR-DXD: 、 MC-SGN-DXD: 、 MsP-SGR-DXD: 、 MsP-YSR-DXD: 、 MsP-SGR-Eciticon: 、 MsP-YSR-Eciticon: 、 MsP-SGR-PABC-Eciticon: 、 MsP-YSR-PABC-Eciticon: 、 MsP-SGE-DXD: 、 MsP-TSR-DXD: 、 MsP-SGK-DXD: 、 MsP-Y(Me)SR-DXD: ,and Mal-PEG8-SGR-PABC-Eciticon: 。 14. The compound according to any one of claims 1-6, wherein, The drug is derived from compounds having the formula (DI) or (D-II): (DI) or (D-II) Rd1 is C 1-6 Alkyl or C 1-6 Alkoxy; Rd2 is a halogen; Or preferably, Rd1 and Rd2 together with the carbon atoms to which they are attached form a 5-10 membered heterocyclic group or a 5-10 membered heteroarylene group, which is optionally surrounded by one, two or three atoms selected from H, D, halogens and C. 1-6 Alkyl substituents; Rd3 is selected from -(CH2). p -ORd a -(CH2) p -C(O)ORd a -(CH2) p -NHC(O)-Rd a and -(CH2) p -CH2NRd b Rd c ; Rd3 is preferably selected from -(CH2). p -ORd a -(CH2) p -C(O)ORd a and -(CH2) p -NHC(O)-Rd a ; Rd a Selected from H, C 1-6 Alkyl, C 1-6 alkylene-OH, C 1-6 Alkylene-NH2, such as H, CH3, CH2OH, CH2CH2OH and CH2CH2NH2; Rd b and Rd c Independently selected from H and C 1-6 Alkyl groups, such as H and CH3; Each p is independently 0, 1, 2, 3, 4, 5, or 6; Alternatively, Rd1 is C 1-6 Alkyl or C 1-6 Alkoxy; Rd2 is a halogen; Or preferably, Rd1 and Rd2 together with the carbon atoms to which they are attached form a 5-6 membered heterocyclic group or a 5-6 membered heteroarylene group (preferably forming a 5-6 membered heterocyclic group), which is optionally surrounded by one, two or three atoms selected from H, D, halogens and C. 1-4 Alkyl substituents; Rd3 is selected from , , , , or Preferably ; Rd a 、Rd b and Rd c Independently selected from H and C 1-6 alkyl; Each p is independently 0, 1, 2, 3, 4, 5, or 6; Alternatively, Rd1 is C 1-4 Alkyl or C 1-4 Alkyl groups, such as Me or OMe; Rd2 is a halogen, such as F; Or preferably, Rd1, Rd2 together with the carbon atoms to which they are attached form , or ; Rd3 is selected from , , , , , , , , , and ; Alternatively, part D of the drug is derived from the following compounds: , , , , , , , , , , , or .

15. The compound of claim 14, wherein, The drug fraction D is selected from compounds having formula (D-III) or (D-IV): (D-III) (D–IV) in, Indicates the attachment site with the rest of the compound; Rd1 and Rd2 are as defined in claim 14; Rd3' is selected from -(CH2) p -ORd a '-、-(CH2) p -C(O)ORd a '-、-(CH2) p -NHC(O)-Rd a '- and -(CH2) p -CH2NRd b Rd c '-; Rd3' is preferably selected from -(CH2). p -ORd a '-、-(CH2) p -C(O)ORd a '- and -(CH2) p -NHC(O)Rd a '-; Rd a 'Selected from key, C 1-6 Alkylene, -C 1-6 alkylene-O- and -C 1-6 Alkylene -NH-, such as bond, -CH2-, -CH2O-, -CH2CH2O- and -CH2CH2NH-; Rd b Selected from H and C 1-6 Alkyl groups, such as H and CH3; Rd c 'Selected from key and C' 1-6 Alkyl groups, such as bonds and -CH2-; Each p is independently 0, 1, 2, 3, 4, 5, or 6; Alternatively, Rd3' is selected from , , , , and ,For example , , , , , , , , and Preferably ; Rd a 'and Rd c 'Independently selected from key and C 1-6 Alkylene; Rd b Selected from H and C 1-6 alkyl; Each p is independently 0, 1, 2, 3, 4, 5, or 6; Alternatively, Part D of this drug is selected from: , , , , , , , , , , , or .

16. The compound according to any one of claims 1-6 and 14-15, wherein the compound is selected from: 、 、 、 、 、 ， in, m, n, P3, P2 and P1 are as defined in any one of claims 1, 3-5 and 11-12; Alternatively, m is selected from 4, 5, 6, 7 or 8, and can be 4, 5 or 8; n is selected from 2, 3 or 4, and can be 3; Alternatively, the compound may be selected from: 。 17. A coupling having formula (II): T-(S’-D) k (II) in, T stands for the target region; S' is a linker, which is a divalent group formed by linking S and T; The linker comprises a peptide-cleavable unit, which comprises a tripeptide having the sequence -P3-P2-P1-, wherein P3, P2 and P1 are as defined in any one of claims 1-16. D represents the drug component; k ranges from 1 to approximately 20.

18. The conjugate of claim 17, wherein the targeting portion is selected from an antibody or its antigen-binding fragment, ligand, or targeting peptide.

19. The conjugate of claim 18, wherein the antibody or its antigen-binding fragment binds to a tumor antigen, a tumor-associated antigen, an immune cell antigen, or a T-cell antigen; The ligand binds to a receptor expressed on tumor cells or immune cells, or to a receptor expressed in the tumor microenvironment; and / or The targeting peptide binds to target molecules expressed on tumor cells or immune cells, or to receptors expressed in the tumor microenvironment.

20. The conjugate of claim 18 or claim 19, wherein the antibody is a monoclonal antibody, a bispecific antibody, or a multispecific antibody.

21. The conjugate according to any one of claims 18-20, wherein the antibody is a mouse antibody, a chimeric antibody, a humanized antibody, or a human antibody.

22. The conjugate according to any one of claims 18-21, wherein the antibody belongs to the isotype selected from the group consisting of IgG, IgA, IgM, IgE and IgD, preferably wherein the antibody belongs to the subtype selected from the group consisting of IgG1, IgG2, IgG3 and IgG4.

23. The conjugate according to any one of claims 18-22, wherein the antigen-binding fragment is selected from the group consisting of: Fab, Fab', F(ab')2, Fv, scFv and ds-scFv.

24. The coupling as described in any one of claims 17-23, wherein, -S'-D is represented by equation (III): (III) in This indicates an attachment site for an antibody or antigen-binding fragment thereof as defined in any one of claims 19-23; L' is represented by the formula -L1'-L2-X-, where L1' is a divalent group formed by connecting L1 and T; L1, L2, P, Y and D are as defined in any one of claims 1-16; Alternatively, L1' is selected from , , and .

25. The coupling as described in any one of claims 17-24, wherein, -S'-D is selected from: ； ； ； ； ； ； ； ； as well as ， in, This indicates the attachment site of the antibody or its antigen-binding fragment as defined in any one of claims 19-23; m, n, P3, P2 and P1 are as defined in any one of claims 1, 3-5 and 11-12; Alternatively, -S'-D is selected from: -MC-VGR-DXD: 、 -MC-SGR-DXD: 、 -MC-VGR-PABC-MMAE: 、 -MC-SGR-PABC-MMAE: 、 -MC-SGC-DXD: 、 -MC-SAC-DXD: 、 -MsP-SGC-DXD: 、 -MC-S(tBu)GC-DXD: 、 -MC-SGOrn(Boc)-DXD: 、 -MC-SGC-PABC-Eciticon: 、 -MC-SGO(DEt)-PABC-Eciticon: 、 -MC-SGE-PABC-Eciticon: 、 -MC-SGE(tBu-PABC-Eciticon: 、 -MC-SGC(Me)-PABC-Eciticon: 、 -MC-SGC(Peg)-PABC-Eciticon: 、 -MC-TGC-PABC-Eciticon: 、 -MC-SGhC(O)-PABC-Eciticon: 、 -MC-YSR-PABC-Eciticon: 、 -MC-SGE-DXD: 、 -MC-SGC(Peg)-DXD: 、 -MC-TSR-DXD: 、 -MC-YSR-DXD: 、 -MC-SGO-DXD: 、 -MC-SGK-DXD: 、 -Mal-PEG4-VGR-DXD: 、 -Mal-PEG4-SGR-DXD: 、 -MC-SGhR-DXD: ,and -MC-SGN-DXD: 、 -MsP-SGR-DXD: 、 -MsP-YSR-DXD: 、 -MsP-SGR-Eciticon: 、 -MsP-YSR-Eciticon: 、 -MsP-SGR-PABC-Eciticon: 、 -MsP-YSR-PABC-Eciticon: 、 -MsP-SGE-DXD: 、 -MsP-TSR-DXD: 、 -MsP-SGK-DXD: 、 -MsP-Y(Me)SR-DXD: ,and -Mal-PEG8-SGR-PABC-Eciticon: ， in, This indicates the attachment site of the antibody or its antigen-binding fragment as defined in any one of claims 19-23.

26. The coupling as described in any one of claims 17-25, wherein, The coupling element is selected from: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 ,and 、 、 、 、 、 、 、 、 、 、 ,and ， Among them, Ab is an antibody or its antigen-binding fragment that binds to tumor antigens, tumor-associated antigens, or immune cell antigens or T cell antigens; And k is an integer selected from 1 to 10, preferably 4 to 8, more preferably k is 4 or 8.

27. The coupling compound according to any one of claims 17-24, wherein -S'-D is selected from: 、 、 、 、 、 ， in, This indicates the attachment site of the antibody or its antigen-binding fragment as defined in any one of claims 19-23; m, n, P3, P2 and P1 are as defined in any one of claims 1, 3-5 and 11-12; Alternatively, -S'-D is selected from: ； in, This indicates the attachment site of the antibody or its antigen-binding fragment as defined in any one of claims 19-23; Alternatively, the coupling element may be selected from: 、 ， Among them, Ab is an antibody or its antigen-binding fragment that binds to tumor antigens, tumor-associated antigens, or immune cell antigens or T cell antigens; And k is an integer selected from 1 to 10, preferably 4 to 8, more preferably k is 4 or 8.

28. The conjugate of claim 26 or 27, wherein the antibody is a Trop2 antibody.

29. A composition comprising the conjugate as described in any one of claims 17-28 and optionally a pharmaceutically acceptable carrier or excipient.

30. The use of any compound of claims 1-16, or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, diastereomer, or mixture thereof, a conjugate of any one of claims 17-28, or a composition of claim 29, in the manufacture of a medicament for treating and / or preventing disease.

31. The compound of any one of claims 1-16, or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, diastereomer, or mixture thereof, a conjugate of any one of claims 17-28, or a composition of claim 29, for use in the treatment and / or prevention of disease.

32. A method for preventing and / or treating a disease in a subject in need, the method comprising administering to the subject the conjugate as described in any one of claims 17-28 or the composition as described in claim 29.

33. The method of claim 32, wherein the method further comprises administering a second therapeutic agent to the subject, preferably wherein the second therapeutic agent is selected from antibodies, chemotherapeutic agents, and small molecule drugs.

34. The use as described in claim 30, or the compound, conjugate, or composition for use as described in claim 31, or the method as described in claim 32 or 33, wherein the disease is selected from cancer, infectious diseases, inflammatory diseases, autoimmune diseases, and immunodeficiency diseases; Preferably, the cancer is selected from head and neck cancer, esophageal cancer, lung cancer, colon cancer, rectal cancer, stomach cancer, pancreatic cancer, ovarian cancer, prostate cancer, breast cancer, leukemia, myeloma, squamous cell carcinoma, melanoma, brain cancer, cervical cancer, liver cancer, bladder cancer, breast cancer, kidney cancer, testicular cancer, and thyroid cancer. Preferably, the autoimmune disease is selected from systemic sclerosis and atherosclerosis.

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