Antibody-drug conjugate and use thereof

Abstract

The present invention relates to an antibody-drug conjugate, and in particular, to an antibody-drug conjugate (ADC) carrying a Ras mutation inhibitor, a composition comprising the ADC molecule, and therapeutic use thereof.

Description

Antibody-drug conjugate and its use Technical Field

[0001] The present disclosure relates to antibody-drug conjugates (ADCs) based on Ras mutation inhibitors, pharmaceutical compositions comprising the same, and methods for their use in treating or preventing diseases associated with Ras mutations. Specifically, the present disclosure relates to compounds, uses, and methods for treating or preventing related diseases such as tumors or cancer by targeting oncogenic mutants of Ras. Background Art

[0002] Ras belongs to the GTPase protein family. Under normal physiological conditions, Ras is activated by growth factors and various other extracellular signals and is responsible for regulating functions such as cell growth, survival, migration, and differentiation. These regulatory functions of Ras are carried out through the conversion between an inactive GDP-bound state and an active GTP-bound state, a "molecular switch." GDP-bound Ras is an inactive form. When exposed to certain pro-growth stimuli such as guanine nucleotide exchange factors (GEFs), it is activated, releasing GDP and binding to GTP, thereby converting to an active GTP-bound state. It recruits and activates various downstream effectors, transmits signals, and thus controls many key cellular processes such as differentiation, survival, and proliferation.

[0003] Ras has GTPase activity, which can cleave the terminal phosphate of GTP and convert it to GDP, that is, convert itself to an inactive state. However, the endogenous GTPase activity of Ras is very low, and the exogenous protein GAP (GTPase activating protein) is required to convert GTP-Ras to GDP-Ras. GAP interacts with Ras and promotes the conversion of GTP to GDP. Therefore, any Ras gene mutation that affects the interaction between Ras and GAP or the conversion of GTP to GDP will cause Ras to be in an over-activated state, thereby continuously transmitting growth and division signals to cells, stimulating continuous cell proliferation, and ultimately leading to tumor formation and development. In fact, dysregulation of the Ras signaling pathway is almost always associated with disease. Over-activating somatic mutations of Ras are one of the most common lesions in human cancer.

[0004] Although mutations in any of the three Ras isoforms (K-Ras, N-Ras, or H-Ras) have been shown to lead to oncogenic transformation, KRas mutations are by far the most common mutations in human cancers, commonly found in pancreatic cancer, lung adenocarcinoma, colorectal cancer, gallbladder cancer, thyroid cancer, and bile duct cancer, and can also be seen in 25% of patients with non-small cell lung cancer. The vast majority of KRas mutations occur at codons G12, G13, and Q61, with approximately 80% of KRas mutations occurring at glycine codon 12, such as G12C, G12D, G12V, G12A, G12R, G12S, and G13D, with G12D being one of the most common mutations.

[0005] In light of this, mutant Ras proteins, such as KRas, have become highly attractive anticancer drug targets in the pharmaceutical field, and the development of inhibitors is considered a very promising research and development direction in anticancer / tumor drug development. However, decades of drug development targeting K-Ras mutations have shown that existing small molecule inhibitors of K-Ras mutations have many difficult-to-overcome drawbacks, such as ineffective delivery to certain lesions such as the colon, breast, and pancreas, severe drug resistance, poor membrane permeability, poor hydrophilicity, unsatisfactory oral pharmacokinetics, and high toxicity following intravenous administration. As a result, KRas has long been considered an "undruggable" target.

[0006] Therefore, the present inventors have been devoted to developing small molecule inhibitors of Ras, such as KRas mutations, with improved structural patterns. Due to the specifically designed structural fragments, these small molecule inhibitors have been shown to have enhanced Ras, such as KRas mutation inhibitory activity and inhibitory activity against related tumors compared to existing Ras, such as KRas mutation inhibitors in the prior art, while having good oral pharmacokinetic properties, thereby having good drugability, reduced toxic side effects, improved drug resistance and safety, and reduced risk of drug interactions.

[0007] Despite this, the field of cancer treatment still urgently needs new therapies that can more accurately deliver Ras, such as KRas mutant inhibitors, to the tumor environment, can be administered in a more friendly way, and prolong the efficacy and reduce toxic side effects and drug resistance. The present disclosure meets the above needs.

[0008] Targeted drug conjugates are a new, highly efficient and low-toxic drug delivery technology that demonstrates significant advantages in the development of anticancer drugs. Cytotoxic molecules are linked to targeting molecules such as antibodies and peptides via suitable linkers. These targeting molecules can selectively deliver the conjugated molecules to tumor tissues that highly express targeting receptors. Through internalization, the anticancer drugs are selectively introduced into cancer cells. The active anticancer molecules are then cut and released using differentially functional molecules enriched in tumor tissues and cancer cells. Because active anticancer molecules can only be released and concentrated within tumor tissues that highly express targeting receptors and specific "cutters," this dual-selective drug delivery method significantly reduces the toxicity of anticancer drugs and improves their efficacy.

[0009] The inventors utilized a targeted conjugation strategy to further link a developed set of Ras, such as KRas, mutant inhibitor compounds to targeting molecules such as antibodies and peptides via suitable linkers, thereby producing a set of ADC compounds based on targeted Ras, such as KRas, mutant inhibitors. The developed ADC compounds, due to the use of targeted Ras, such as K-Ras enzyme inhibitors, as ADC payloads, exhibit dual selectivity and significantly reduced toxicity. Specific modifications to the linker units enhance coupling efficiency and product purity, inhibit ADC macromolecular aggregation, and improve drug efficacy. Furthermore, these ADC macromolecules delay the metabolism of the loaded Ras, such as KRas, mutant inhibitor, further improving the drug's systemic stability and PK properties. This results in a longer in vivo half-life, sustained target inhibition, and prolonged drug efficacy, thereby delaying the development of drug resistance.

[0010] SUMMARY OF THE INVENTION

[0011] To meet the above needs in the art, the present inventors conducted in-depth research and coupled a group of Ras-targeting, such as KRas mutation inhibitor compounds, with target molecules through selected linker units to obtain antibody-drug conjugates as shown in the examples.

[0012] Currently under development, Ras inhibitors, such as KRas inhibitors, have shown many defects during the development and use process, including ineffective delivery to certain lesions, severe drug resistance, poor membrane permeability, poor hydrophilicity, poor oral PK properties, high toxicity after intravenous administration, etc.; the antibody-drug conjugates disclosed herein, by innovative structural modifications to the components of the linker unit of the ADC and using the linker unit for the first time to conjugate Ras small molecule inhibitors and antibody-directed units, can be quickly and effectively internalized by tumor cells, and have demonstrated significant tumor growth inhibition activity and good tolerability compared to single antibodies and single Ras inhibitors in animal models, thus overcoming the above defects. It is expected that more mature and friendly administration methods (such as oral and parenteral) can be adopted to provide enhanced and prolonged efficacy, good pharmacokinetic properties, reduced toxic side effects and drug resistance, as well as expanded indications and beneficiary populations.

[0013] Thus, in a first aspect, the present disclosure provides an antibody-drug conjugate (ADC) having the following formula (X) or a pharmaceutically acceptable salt or solvate thereof: [PL] q -Ab (X)

[0014] in,

[0015] P represents a Ras inhibitor, such as a Ras inhibitor as defined in the P unit portion of the drug disclosed herein (such as a compound of formula (I) and its various subformulas as defined in the disclosure);

[0016] L represents the linker unit that connects P to Ab;

[0017] q represents the number of [PL] linkers attached to Ab, for example, q = an integer or non-integer from 1 to 20, such as 1-10, 1-8, 2-8, 3-8, 4-8 or 6-8;

[0018] Ab stands for antibody or antigen-binding fragment.

[0019] In a second aspect, the present disclosure provides a pharmaceutical composition comprising an ADC of the present disclosure or a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof, optionally at least one other therapeutic agent, and optionally one or more pharmaceutically acceptable excipients.

[0020] In a third aspect, the present disclosure provides an ADC of the present disclosure or a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising the same, for use as a therapeutic agent for treating or preventing diseases mediated by Ras mutant proteins (such as, but not limited to, G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, and G13D mutant proteins), more specifically for treating or preventing hyperproliferative diseases, especially as an anti-tumor therapeutic agent. In some specific embodiments, the Ras mutant protein is a KRas mutant protein.

[0021] In a fourth aspect, the present disclosure provides the use of the ADC of the present disclosure or its stereoisomers, tautomers, stable isotopic variants, pharmaceutically acceptable salts or solvates, or pharmaceutical compositions comprising the same, in preventing or treating diseases mediated by Ras mutant proteins (such as, but not limited to, G12C mutations, G12D mutations, G12V mutations, G12A mutations, G12R mutations, G12S mutations and G13D mutant proteins), more specifically in treating or preventing hyperproliferative diseases, especially tumors.

[0022] In a fifth aspect, the present disclosure provides a method for treating or preventing a disease mediated by a Ras mutant protein (such as, but not limited to, a G12C mutation, a G12D mutation, a G12V mutation, a G12A mutation, a G12R mutation, a G12S mutation, and a G13D mutant protein) in a subject, the method comprising administering to a human or animal an ADC of the present disclosure, or a stereoisomer, tautomer, stable isotopic variant, a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising the same; specifically, the present disclosure provides a method for treating or preventing a hyperproliferative disease, especially a tumor, in a subject, the method comprising administering to a human or animal an ADC of the present disclosure, or a stereoisomer, tautomer, stable isotopic variant, a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising the same.

[0023] In a sixth aspect, the present disclosure provides use of the ADC of the present disclosure or its stereoisomers, tautomers, stable isotopic variants, pharmaceutically acceptable salts or solvates, or pharmaceutical compositions comprising the same, in the preparation of medicaments for preventing or treating diseases mediated by Ras mutant proteins (such as, but not limited to, G12C mutations, G12D mutations, G12V mutations, G12A mutations, G12R mutations, G12S mutations, and G13D mutant proteins); specifically, the present disclosure provides use of the ADC of the present disclosure or its stereoisomers, tautomers, stable isotopic variants, pharmaceutically acceptable salts or solvates, or pharmaceutical compositions comprising the same, in the preparation of medicaments for treating or preventing hyperproliferative diseases, especially tumors.

[0024] In a seventh aspect, the present disclosure provides a drug combination comprising an ADC of the present disclosure or a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof, and at least one other therapeutic agent; the combination is used for preventing or treating diseases mediated by Ras mutant proteins (such as, but not limited to, G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation and G13D mutant proteins), more specifically for treating or preventing hyperproliferative diseases, especially tumors; and a method for treating or preventing diseases mediated by Ras mutant proteins (such as G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation and G13D mutant proteins), more specifically hyperproliferative diseases, especially tumors in a subject, the method comprising administering the drug combination of the present disclosure to a human or animal.

[0025] In an eighth aspect, the present disclosure further provides a method for preparing an ADC of the present disclosure or a stereoisomer, tautomer, stable isotope variant, pharmaceutically acceptable salt or solvate thereof by conjugating a Ras inhibitor as defined herein to an antibody or antigen-binding fragment thereof via a linker unit. The Ras inhibitor compound as defined herein can be conjugated to an antibody or antigen-binding fragment via a cleavable or non-cleavable linker unit. In specific embodiments, upon cleavage of the linker unit, the Ras inhibitor is released into tumor cells, cancer-associated immune cells or the tumor microenvironment.

[0026] In some embodiments of the various aspects of the present disclosure above, the Ras mutant protein is a KRas mutant protein, specifically a KRas G12D mutant protein, and accordingly the P inhibitor compound is a KRas inhibitor, specifically a KRas G12D inhibitor; in other embodiments, the Ras mutant protein is not limited to a specific subtype and / or mutation site, that is, it is pan-Ras, and accordingly the P inhibitor compound is a pan-Ras inhibitor.

[0027] The present disclosure is further illustrated in the following drawings and specific embodiments. However, these drawings and specific embodiments should not be considered to limit the scope of the present disclosure, and modifications that are readily apparent to those skilled in the art will be included within the spirit of the present disclosure and the scope of protection of the appended claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] FIG1 shows the anti-tumor effects and body weight changes of some representative ADC compounds of the present disclosure in a KRAS-G12D mutant human colon cancer cell GP2D subcutaneous xenograft BALB / c Nude mouse animal model (dosage: 20 mg / kg, administration on D0 / 7, for 21 days).

[0029] Figure 2 shows the anti-tumor effects and body weight changes of some representative ADC compounds of the present disclosure in a subcutaneous xenograft BALB / c Nude mouse animal model of KRAS-G12D mutant human colon cancer cell GP2D in another batch of experiments (dosage: 10 mg / kg, administration on D0, for 21 days).

[0030] FIG3 shows the anti-tumor effects of some representative ADC compounds of the present disclosure in another batch of experiments in a BALB / c Nude mouse subcutaneous xenograft model of human colon cancer cells GP2D with KRAS-G12D mutation (dosage: 10 mg / kg, administration on D0, for 20 days).

[0031] Figures 4A, 4B, and 4C show the results from another batch of experiments, namely, the anti-tumor effects of some representative ADC compounds of the present disclosure in a KRAS-G12D mutant human colon cancer cell GP2D subcutaneous xenograft BALB / c Nude mouse animal model (dosage: 10 mg / kg, dosing on D0, for 20 days). For the purpose of clear display, the example compounds in this batch of experiments are separated and shown in Figures 4A, 4B, and 4C, respectively.

[0032] Figures 5 and 6 respectively show the anti-tumor effects of some representative ADC compounds, drug P units and antibodies of the present disclosure in a subcutaneous xenograft BALB / c Nude mouse animal model of KRAS-G12D mutant human colon cancer cell GP2D in another batch of experiments (dosage of 10 mg / kg, dosing on D0 and / or 7 and / or 14 days, for 21 days).

[0033] Detailed Description of the Invention

[0034] definition

[0035] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. For the purposes of this disclosure, the following terms are defined below.

[0036] When a trade name is used herein, unless the context indicates otherwise, the trade name includes the product formulation, generic drug, and active pharmaceutical ingredient of the trade name product.

[0037] As used herein, the term "substantially" refers to a majority, i.e., > about 50% of a population, mixture, sample, content or any other numerical value, preferably greater than about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%.

[0038] In this document, the term "and / or" should be understood to mean any one of the options or a combination of any two or more of the options.

[0039] As used herein, the term "ADC" or "conjugate" refers to an antibody-drug conjugate.

[0040] As used herein, the term "drug" refers to a substance that produces a beneficial preventive or therapeutic effect on diseases mediated by Ras mutant proteins, such as KRas mutant proteins (such as, but not limited to, G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation, and G13D mutant proteins).

[0041] As used herein, the term "Ras mutant" or "Ras mutant protein" refers to a protein encoded and expressed by a Ras gene in which one or more codons are mutated, typically including but not limited to a Ras protein, such as a mutant HRas, NRas, or KRas, in which a glycine residue at codon 12, a glycine residue at codon 13, or a glutamine residue at codon 61 of Ras is mutated. These residues are located in the active site of Ras, and their mutations can impair the intrinsic or GAP-catalyzed GTPase activity of Ras, resulting in the persistence of Ras bound to GTP.

[0042] For the purposes of this disclosure, "Ras mutation" or "Ras mutant protein" and "Ras" when describing inhibitory activity are used interchangeably, for example to refer to mutant KRas, such as but not limited to KRas-G12C (glycine to cysteine ​​mutation at codon G12), KRas-G12D (glycine to aspartic acid mutation at codon G12), HRas-G12D, NRas-G12D, KRas-G12V (glycine to valine mutation at codon G12), ... 3D (glycine to aspartic acid mutation at codon G13); in some embodiments, refers to a KRas mutant protein, more particularly refers to a KRas-G12C mutant protein, a KRas-G12D mutant protein, a KRas-G12V mutant protein, a G12A mutant protein, a G12R mutant protein, a G12S mutant protein, a KRas-G13D mutant protein, most particularly refers to KRas-G12D, and in other embodiments refers to a pan-RAS mutant protein, i.e., not limited to a specific subtype and mutation site.

[0043] As used herein, the term "Ras mutation-mediated disease" refers to a disease in which Ras mutations contribute to the onset and progression of the disease, or in which inhibition of Ras mutations reduces the incidence of the disease, reduces, or eliminates the symptoms of the disease. For purposes of this disclosure, "Ras mutation-mediated disease" refers in some embodiments to a KRas mutation-mediated disease, most preferably KRas-G12D, and in other embodiments to a pan-Ras-mediated disease, such as a hyperproliferative disease such as cancer or a tumor.

[0044] As used herein, the term "cancer" or "tumor" refers to abnormal cell growth and proliferation, including solid tumors and blood-borne tumors, whether malignant or benign, and all precancerous and cancerous cells and tissues. For various aspects of the present disclosure, the cancer or tumor includes, but is not limited to, lung adenocarcinoma, lung cancer (including squamous cell carcinoma of the lung and non-small cell lung cancer, small cell lung cancer (SCLC)), bone cancer, pancreatic cancer, pancreatic ductal adenocarcinoma, skin cancer, head and neck cancer (including head and neck squamous cell carcinoma), melanoma (including skin or intraocular melanoma), squamous cell carcinoma, cancer of the anal region, testicular cancer, urethral cancer, ureteral cancer, penile cancer, prostate cancer (including hormone-refractory prostate cancer), bladder cancer, uterine cancer, Ovarian cancer, ovarian epithelial cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, stomach cancer, gastric adenocarcinoma, colon cancer, rectal cancer, colorectal cancer, liver cancer, breast cancer (including metastatic breast cancer and triple-negative breast cancer (TNBC)), esophageal cancer, small intestine cancer, lip cancer, laryngeal cancer, nasopharyngeal cancer, oral cancer, salivary gland cancer, peritoneal cancer, gastrointestinal stromal tumor, gastroesophageal junction (GEJ) cancer, mesothelioma, biliary tract cancer, hepatoma, seminoma, soft tissue sarcoma, osteosarcoma, urothelial cancer Cancer, sweat gland cancer, endocrine system cancer, thyroid cancer, medullary thyroid cancer, follicular thyroid cancer, papillary thyroid cancer, parathyroid cancer, kidney cancer, renal parenchymal cancer, renal cell carcinoma, renal pelvis cancer, adrenal gland cancer, brain cancer such as glioblastoma, astrocytoma, meningioma, medulloblastoma, peripheral neuroectodermal tumor, glioblastoma (including glioblastoma multiforme), neuroblastoma; chronic or acute leukemia, Hodgkin's disease, lymphoma (including lymphocytic lymphoma, Hodgkin's lymphoma) , non-Hodgkin lymphoma, Burkitt lymphoma, adult T-cell lymphoma, diffuse large-scale lymphoma (DLBCL), primary CNS lymphoma), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CLL) and lymphocytic carcinoma, acute myeloid leukemia (AML), myeloid leukemia (chronic myeloid leukemia (CML), central nervous system tumors (CNS), spinal tumors, brainstem gliomas, or pituitary adenomas.

[0045] For various aspects of the present disclosure, preferably, the cancer or tumor is associated with a Ras mutation, such as a KRas mutation, including but not limited to the above-mentioned tumor types and their preferred ranges. Particularly preferred tumors of the present invention include lung cancer, lung adenocarcinoma, colon cancer, rectal cancer, pancreatic cancer, endometrial cancer, bile duct cancer, leukemia and ovarian cancer.

[0046] As used herein, the term "anti-tumor effect" refers to a biological effect that can be characterized in various ways, including but not limited to, for example, a decrease in tumor volume, a decrease in the number of tumor cells, a decrease in tumor cell proliferation, or a decrease in tumor cell survival.

[0047] As used herein, the terms "inhibit" and "reduce" or any variants of these terms refer to the ability of a bioactive agent to reduce the signaling activity of a target of interest by interacting directly or indirectly with the target, and refer to any measurable reduction or complete inhibition of the activity of a target of interest. For example, compared to normal, the activity (e.g., Ras activity, e.g., KRas activity) can be reduced by about, up to about, or at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99% or more, or any range derivable therein.

[0048] As used herein, the term "selective inhibition" refers to the ability of a bioactive agent to preferentially reduce the signaling activity of a target over off-target signaling activity by directly or indirectly interacting with the target. With respect to the Ras inhibitors and ADCs disclosed herein, the Ras inhibitors and ADCs thereof have the ability to selectively inhibit G12 or G13 mutations of the KRas protein, such as G12C mutations, G12D mutations, G12V mutations, G12A mutations, G12R mutations, G12S mutations, and G13D mutations, with respect to various mutations occurring in one or more codons of the Ras protein. Preferably, the inhibitors selectively inhibit the G12D mutation of the KRas protein. For example, the Ras inhibitors and ADCs thereof of the present disclosure may have at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or more, or any range derivable therein, against a particular mutation, such as KRas-G12D, as compared to another particular Ras mutation, or at least 0.1-, 0.5-, 1-, 2-, 3-, 4-, 5-, 10-, 25-, 50-, 100-, 250-, or 500-fold or more of the inhibitory activity against a particular mutation, such as KRas-G12D, as compared to another specific Ras mutation.

[0049] The term "dual selective inhibition" used in this article refers to the ability of the conjugated drug to, on the one hand, target and inhibit specific Ras mutations through small molecule targeted inhibitors, and on the other hand, to highly selectively deliver the loaded Ras targeted inhibitor target to tumor tissues with high expression of targeting receptors through the connected targeting antibody molecules, thereby exerting a dual selective inhibitory effect.

[0050] As used herein, the term "antigen" refers to an entity that specifically binds to an antibody.

[0051] As used herein, the term "antibody" refers to a polypeptide comprising at least a light chain or heavy chain immunoglobulin variable region that specifically recognizes and binds to an antigen. The term encompasses various antibody structures, including but not limited to monoclonal antibodies, single-chain antibodies or multi-chain antibodies, monospecific or multispecific antibodies (e.g., bispecific antibodies), chimeric or humanized antibodies, full-length antibodies, and antibody fragments, as long as they exhibit the desired antigen-binding activity. Antibodies can be of any class (e.g., IgG, IgE, IgM, IgD, and IgA), type (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subtype.

[0052] As used herein, the terms "antibody fragment" and "antigen-binding fragment" of an antibody are used interchangeably and refer to molecules that are not complete antibodies, but comprise portions of an intact antibody that bind to the antigen to which the intact antibody binds. As will be appreciated by those skilled in the art, for antigen binding purposes, antibody fragments typically comprise amino acid residues from "complementarity determining regions" or "CDRs." Antibody fragments can be prepared by recombinant DNA techniques, or by enzymatic or chemical cleavage of intact antibodies. Antigen-binding fragments include, but are not limited to, Fab, scFab, Fab', F(ab')2, Fab'-SH, Fv, single-chain Fv, diabodies, triabodies, tetrabodies, minibodies, single-domain antibodies (sdAbs), and multispecific antibodies formed from antibody fragments.

[0053] Herein, reference to an IgG antibody means that the antibody is a heterotetrameric protein having an IgG-class immunoglobulin structure. In an IgG antibody, typically the VH-CH1 of the heavy chain pairs with the VL-CL of the light chain to form a Fab fragment that specifically binds to an antigen. Thus, an IgG antibody essentially consists of two Fab molecules connected by an immunoglobulin hinge region and two dimerized Fc regions. In some embodiments, an IgG antibody is, for example, an IgG1, IgG2, IgG3, or IgG4 antibody. In other embodiments, an IgG antibody is an IgGκ or IgGλ antibody, for example, an IgG1κ or IgG1λ antibody.

[0054] As used herein, the terms "complementarity determining region" or "CDR region" or "CDR" or "hypervariable region" are used interchangeably and refer to regions in an antibody variable domain that are highly variable in sequence and form structurally defined loops ("hypervariable loops") and / or contain antigen contact residues ("antigen contact points"). CDRs are primarily responsible for binding to antigenic epitopes. As used herein, the CDRs of the antibody heavy and light chains are numbered sequentially from the N-terminus and are commonly referred to as CDR1, CDR2, and CDR3. The CDRs located within the antibody heavy chain variable domain are also referred to as HCDR1, HCDR2, and HCDR3, while the CDRs located within the antibody light chain variable domain are referred to as LCDR1, LCDR2, and LCDR3. In a given light chain variable region or heavy chain variable region amino acid sequence, the CDR sequence can be determined using a variety of methods well known in the art, including CDR sequences defined based on Kabat, AbM, Chothia, Contact, and IMGT. In addition, CDRs can also be determined based on having the same Kabat numbering position as a reference CDR sequence.

[0055] As used herein, a "variable region" or "variable domain" is a domain in the heavy or light chain of an antibody that is involved in the binding of an antibody to its antigen. The heavy chain variable region (VH) and light chain variable region (VL) can be further divided into hypervariable regions (HVRs, also known as complementarity determining regions (CDRs)), interspersed with relatively conserved regions (i.e., framework regions (FRs)). Each VH and VL consists of three CDRs and four FRs, arranged in the following order from amino terminus to carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In some aspects, antibody variable regions can be modified by CDR grafting. Since CDR sequences are responsible for most antibody-antigen interactions, recombinant antibody variants that mimic the properties of known antibodies can be constructed. In this antibody variant, the CDR sequences from a known antibody are grafted onto the framework regions of a different antibody with different properties. The properties of the mutated and / or modified antibodies or ADC conjugates comprising the same can be evaluated in in vitro or in vivo assays, such as target antigen binding properties or other desired functional properties, such as ADC internalization, pharmacokinetics, and in vivo tumor killing activity.

[0056] As used herein, the term "isotype" refers to the antibody type determined by the constant region of the antibody heavy chain. For example, the antibody portion of an ADC according to the present disclosure can be an IgA (e.g., IgA1 or IgA2), IgG1, IgG2 (e.g., IgG2a or IgG2b), IgG3, IgG4, IgE, IgM, and IgD antibody, and has a heavy chain constant region of the immunoglobulin type. In addition, the present disclosure contemplates not only antibodies employing native sequence constant regions, but also antibodies comprising variant sequence constant regions.

[0057] As used herein, the term "epitope" includes any protein determinant capable of specifically binding to an immunoglobulin or otherwise interacting with a molecule. Epitope determinants typically consist of chemically active surface groups of molecules, such as amino acids or carbohydrates or sugar side chains, and may have specific three-dimensional structural characteristics as well as specific charge characteristics. Epitopes may be "linear" or "conformational." The difference between conformational and linear epitopes is that binding to the former, but not the latter, is lost in the presence of denaturing solvents.

[0058] As used herein, the term "receptor-mediated endocytosis" refers to the process by which a ligand / receptor complex is internalized and delivered to the cytosol or translocated to an appropriate intracellular compartment, triggered by the binding of a ligand to a corresponding receptor on the cell surface. The receptor-mediated endocytosis activity of an antibody can be characterized by measuring the endocytosis rate.

[0059] In this article, " sequence identity " refers to the identical degree of sequence based on Nucleotide or amino acid by Nucleotide in a comparison window. " sequence identity percentage ratio " can be calculated in the following manner: the sequence of two optimal comparisons is compared in a comparison window, determine that there is identical nucleic acid base (for example A, T, C, G, I) or identical amino acid residue (for example, Ala, Pro, Ser, Thr, Gly, Val, Leu, Ile, Phe, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, Gln, Cys and Met) position in two sequences to obtain the number of matching positions, the number of matching positions is divided by the total number of positions (that is, window size) in the comparison window, and the result is multiplied by 100, to produce the sequence identity percentage ratio. For determining the optimal comparison carried out of sequence identity percentage ratio, can realize by several ways known in the art, for example, use publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared or over a region of interest.

[0060] As used herein, the term "isolated" antibody is one that has been separated from the components of its natural environment. In some embodiments, the antibody is purified to greater than 90%, 95%, or 99% purity, which can be determined by, for example, electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reversed-phase HPLC).

[0061] As used herein, the term "affinity" or "binding affinity" refers to the intrinsic binding affinity that reflects the interaction between members of a binding pair, such as the strength of the interaction between an antibody and an antigen at a single antigenic site. The stronger the interaction, the stronger the affinity. The affinity of a molecule for its partner can generally be expressed by the equilibrium dissociation constant (K D ) represents the equilibrium dissociation constant, which is the dissociation rate constant and the association rate constant (k dis and k on Affinity can be measured by common methods known in the art, such as ELISA assay based on antigen protein or cells, flow cytometry, biofilm optical interferometry (BLI) technology, etc.

[0062] In this article, the term "K D "(M)" in this article refers to the dissociation equilibrium constant of a specific antibody-antigen interaction. Affinity and K D The values ​​are inversely correlated, that is, the higher the affinity, the higher the K D The smaller the value, the lower the affinity. D The larger the value. Generally, K D The value depends on the dissociation rate constant (Kd or Kdis, sec -1 ) and the binding rate constant (Ka, M -1 ×sec -1 ).

[0063] As used herein, the term "binding" or "specific binding" refers to the ability of a single antibody combining site to react with one antigenic determinant but not with a different antigenic determinant. -6 M or smaller K D Value affinity binding, e.g., K D The value is about 10 -7 M or smaller, or about 10 -8 M or less. Binding K to nonspecific antigens (e.g., irrelevant antigens such as BSA) D The K value of the antibody binding to its cognate antigen is compared D Value, preferably at least 100 times or for example at least 1000 times lower. D The measurement of the value is known in the art, for example based on the biofilm layer interferometry (BLI) technique, for example in ForteBio In the instrument, the measurement is performed using antibodies as ligands and antigens as analytes.

[0064] As used herein, the term "effector function" refers to those biological activities attributable to the Fc region of an antibody, which vary with the antibody class. It is known that the IgG Fc region can mediate several important effector functions, such as cytokine induction, ADCC, phagocytosis, complement-dependent cytotoxicity (CDC), and the half-life / clearance rate of antibodies and antigen-antibody complexes. In some cases, depending on the therapeutic purpose, these effector functions are ideal for therapeutic antibodies, but may be unnecessary in other cases. Therefore, in one embodiment, the present disclosure provides antibodies having an Fc region that induces effector functions such as ADCC or CDC, thereby inducing tumor cell apoptosis, cell lysis, and / or inhibiting the proliferation, dissemination and / or metastasis of tumor cells carrying the antigen TF in tumor cells carrying the TF antigen. In other embodiments, the present disclosure also provides antibodies having an Fc region with altered effector functions. The effector functions can be altered by making sequence changes to the Fc region of the antibody. Alternatively, antibodies having an altered type of glycosylation in the Fc region can be prepared. Alteration of the glycosylation pattern of the Fc region can be conveniently accomplished by altering the amino acid sequence of the Fc region so as to create or remove one or more glycosylation sites.

[0065] As used herein, the term "Linker unit" or "Linker" refers to a bifunctional moiety that connects a drug to an antibody in an antibody-drug conjugate. The Linker unit of the present disclosure has multiple components, such as a self-immolative linker, a cleavable linker, a property-modifying unit, and an antibody linker.

[0066] As used herein, the term "self-immolative linker" refers to a temporary extender, spacer, or placeholder unit that connects two or more molecules together through chemical bonds that break under defined conditions to release the two molecules. Typically, a self-immolative linker can be linear or branched, and can connect two or more identical molecules together, or can connect two or more different molecules together. A self-immolative unit can be defined as a bifunctional chemical group that is capable of covalently linking two spaced chemical moieties together to form a generally stable molecule, releasing one of the spaced chemical moieties from the molecule by enzymatic cleavage; and, after the enzymatic cleavage, spontaneously cleaving from the remaining portion of the bifunctional chemical group to release the other of the spaced chemical moieties. In some embodiments, a self-immolative unit refers to a heterocyclic self-immolative moiety. Typical self-immolative linkers include, but are not limited to, His-Ala, p-aminobenzyloxycarbonyl (PABC), p-hydroxybenzyloxycarbonyl, 2,4-bis(hydroxymethyl)aniline, -NH-(CH2)4-C(O)-, and -NH-(CH2)3-C(O)-.

[0067] As used herein, the term "cleavable linker" refers to a portion of the linker unit of an ADC that is unstable in vivo. Preferably, a "cleavable linker" allows activation of a marker or therapeutic agent by cleaving the marker or agent from the rest of the conjugate. Operationally defined, a cleavable linker is preferably cleaved by the biological environment in vivo. Cleavage can be from any process without limitation, such as enzymatic, reduction, pH, etc. Preferably, the cleavable group is selected so that activation occurs at the desired site of action, which can be a site in or near a target cell (e.g., a cancer cell) or tissue, such as a therapeutic or marker active site. This cleavage can be enzymatic, and exemplary enzymatic cleavage groups include natural amino acids or peptide sequences ending in natural amino acids, and are attached to a linker unit or a self-degrading linker at its carboxyl terminus.

[0068] As used herein, the term "antibody linker" refers to any chemical group designed to facilitate the attachment of a drug conjugate to an antibody.

[0069] As used herein, the term "property-modifying unit" refers to a functional moiety attached to the linker unit of an ADC in a serial or branched manner, which is intended to modulate the properties of the ADC, such as stability in the blood circulation, improved hydrophilicity, etc. Commonly used property-modifying units of ADCs include, but are not limited to, polyethylene glycol (PEG), hydrophilic peptides, monosaccharides, oligosaccharides, polysaccharides, cyclodextrin units, polyamines, polyamides, dendrimers, and bifunctional hydrocarbon chains. In the ADCs of the present disclosure, the property-modifying unit can be attached to the linker unit as a separate component in a serial manner, or as a branched chain of each component in the linker unit, for example, the property-modifying unit can be attached to a self-immolative linker, a cleavable linker, and / or an antibody linker.

[0070] In this article, specific examples of the term "property-modulating unit" include "solubilizing sugar unit", which refers to a sugar unit connected to a linker unit, such as a self-degrading linker portion of a linker (directly connected or indirectly connected through other structural fragments). The sugar unit, for example, together with the self-degrading linker, is specifically hydrolyzed in vivo by enzymes such as β-glucuronidase or β-galactosidase, and releases the drug payload through a self-degradation reaction. In the antibody-drug conjugates of the present disclosure, the introduction of a sugar group into the linker unit helps to improve the hydrophilicity of the conjugate chain, increase the load rate to the theoretical maximum value, and at the same time improve the solubility and purity of the conjugate, reduce aggregation, improve the drugability of the conjugate, and ultimately enhance the effect of inhibiting tumor cell proliferation.

[0071] In this article, "monosaccharide" as a property-modifying unit refers to polyhydroxy aldehydes (aldose) or polyhydroxy ketones (ketose) containing 3 or more carbon atoms and their derivatives. It is the basic structural unit of carbohydrates and their complexes, which can no longer be hydrolyzed and is not connected to other similar units by glycosidic bonds. According to the number of carbon atoms in the monosaccharide, monosaccharides can also be divided into three-carbon sugars (triose), four-carbon sugars (tetraose), five-carbon sugars (pentose), six-carbon sugars (hexose), seven-carbon sugars (heptose), etc., and the ADC disclosed herein preferably carries five-carbon sugars or six-carbon sugars. Derivatives of the monosaccharide include, for example, phosphate esters, sugar alcohols, sugar acids, deoxy sugars, amino sugars, acylated amino sugars and glycosides of monosaccharides. Examples of monosaccharides or their derivatives include, but are not limited to, glyceraldehyde, glyceraldehyde phosphate, dihydroxyacetone phosphate, erythrose, erythrulose, threose, arabinose, ribose, ribulose, deoxyribose, ribitol, ribose phosphate, xylose, xylulose, xylitol, lyxose, glucose, glucosamine, acetylglucosamine, glucuronic acid, acetylglucosamine acid, glucose phosphate, mannose, mannitol, aminomannitol, acetylglucosamine, fructose, fructose phosphate, galactose, galactitol, aminogalactose, acetylgalactosamine, allose, deoxyallose, altrose, deoxyaltrose, chinchonose, rhamnose, psicose, sorbose, sorbitol, tagaose, gulose, deoxygulose, idose, talose, fucose, deoxytalose, and the like.

[0072] It should be noted that the monosaccharides and their derivatives described in the present disclosure include their D configuration, L configuration, racemate (DL) and meso form (meso), and also include any optically active form ((+), (-), (±)). The monosaccharides and their derivatives described in the present disclosure include their chain isomers, cyclic isomers or mixtures thereof, wherein the cyclic form includes α-type anomer and β-type anomer, and, for example, includes pyranose form or furanose form.

[0073] In this article, "disaccharide" as a property-modifying unit refers to a compound and its derivatives formed by two monosaccharides connected by a glycosidic bond. It is divided into two types according to the different connection methods: one is a non-reducing sugar formed by dehydrating the hemiacetal (hemiaketal) hydroxyl groups of two sugars into a glycoside, which is a glycosyl glycoside; the other is a reducing sugar formed by the hemiacetal (hemiaketal) hydroxyl group of one sugar and the non-hemiaacetal (hemiaketal) hydroxyl group of another sugar into a glycoside, which is a glycosyl sugar. The glycosidic bond can be an α-1,4 glycosidic bond, an α-1,6 glycosidic bond, a β-1,4 glycosidic bond, an α-1-2β glycosidic bond, a β-1,6 glycosidic bond, an α-1,1 glycosidic bond, a β-1,3 glycosidic bond, or a β-2,1 glycosidic bond. Specific examples of disaccharides include but are not limited to maltose, isomaltose, lactose, sucrose, chitobiose, rutinose, trehalose, xylobiose, gentiobiose, etc., wherein the monosaccharide units constituting the disaccharide can be in the form of various monosaccharide derivatives as described above.

[0074] As used herein, the term "oligosaccharide" refers to low-polymer sugars formed by 3 to 9 monosaccharides connected by glycosidic bonds and their derivatives, wherein the monosaccharides, glycosidic bonds and derivatives are as defined above, for example but not limited to mannotriose, selaginose, gentiotriose, plantaginose, stachyose, raffinose, etc.

[0075] As used herein, the term "polysaccharide" refers to a group of 10 or more monosaccharide groups linked by glycosidic bonds, which may be α-type, β-type, or a mixed α / β type, and may be linear, branched, or cyclic in structure. A polysaccharide may be a homopolysaccharide composed of a single monosaccharide, wherein the sugar units of the homopolysaccharide are selected from triose, tetraose, pentose, hexose, heptose, and octose, or deoxysugar units, such as dextran, xylan, and polysialic acid; or a heteropolysaccharide composed of two or more monosaccharides, such as hyaluronic acid and heparin. Examples of polysaccharides include, but are not limited to, dextran, levulin, hyaluronic acid, cyclodextrin (α, β and γ, etc.), hydroxyethyl starch, xylan, water-soluble starch, water-soluble cellulose, carboxymethyl cellulose, aminogalactose, polysialic acid, rhamnose, ganoderma lucidum polysaccharide, lentinan, chitin, chitosan, alginate, carrageenan, gellan gum, pullulan, scleroglucan, xanthan gum, xyloglucan, amylose, etc. The polysaccharide may have, or be modified to have, one or more substituents selected from the group consisting of carboxylic acid group, carboxylate group, amino group, sulfonic acid group, sulfonate group, phosphate group, phosphate group, hydroxyethyl group, hydroxypropyl group, methyl group, acyl group, carboxymethyl group, natural amino acid group, non-natural amino acid group, etc.

[0076] In this document, the term "derivative" used to describe sugar units mainly refers to sugar phosphates (sugar compounds formed by esterification of one or more hydroxyl groups of a sugar with phosphoric acid), sugar alcohols (sugar compounds formed by reducing the carbonyl group of a sugar to OH), sugar acids (aldonic acids formed by oxidation of the aldehyde group of an aldose to a carboxyl group, uronic acids formed by oxidation of the primary alcohol group of an aldose to a carboxyl group, and saccharic acids in which both the aldehyde group and the primary alcohol group of an aldose are oxidized to carboxyl groups), deoxysugars (sugars in which one or two hydroxyl groups of a sugar molecule are replaced by hydrogen atoms), amino sugars (sugar derivatives in which one or more hydroxyl groups of a sugar molecule are replaced by amino groups), acylated amino sugars (sugar derivatives in which the amino group of an amino sugar is acylated), and glycoside forms (sugar-containing derivatives formed by condensation of a hemiacetal hydroxyl group of a sugar with a hydroxyl group, an amine group, or a thiol group of another molecule such as an alcohol, a sugar, a purine, or a pyrimidine, i.e., derivatives in which a sugar residue (sugar minus the hemiacetal hydroxyl group) and a ligand are linked by a glycosidic bond), preferably in the form of sugar acids, amino sugars, or acylated amino sugars.

[0077] Sugar derivatives suitable for ADCs of the present disclosure may also be derivatives formed by derivatizing sugar molecules in more than one of the above ways, such as acylated aminouronic acids, such as 2-acetylamino-2-deoxy-D-galacturonic acid, 2-acetylamino-2-deoxy-D-glucuronic acid, and 2-acetylamino-2-deoxy-D-mannuronic acid.

[0078] In this article, specific examples of the term "property-modulating unit" also include "PEG units". The term "PEG unit" refers to an organic portion comprising repeated ethyleneoxy subunits (PEG or PEG subunits), which can be polydisperse, monodisperse or discrete (i.e., having a discrete number of ethyleneoxy subunits). Polydisperse PEGs are heterogeneous mixtures of size and molecular weight, while monodisperse PEGs are typically purified from heterogeneous mixtures and therefore have a single chain length and molecular weight. Preferred PEG units include discrete PEGs, which are compounds synthesized in a stepwise manner rather than via a polymerization process. Discrete PEGs provide single molecules with defined and specified chain lengths.

[0079] The PEG unit provided herein includes one or more polyethylene glycol chains, each polyethylene glycol chain consisting of one or more alkene ethyleneoxy subunits, which are covalently linked to each other. The polyethylene glycol chains can be linked together, for example, in a linear, branched or star-shaped configuration. Typically, before being incorporated into the ADC conjugate, at least one polyethylene glycol chain is derived from an alkyl moiety substituted with an electrophilic group at one end to covalently link to the carbamate nitrogen of the methylene carbamate unit. Typically, the terminal ethyleneoxy group that does not participate in the covalent connection to the remainder of the linker unit in each polyethylene glycol chain is modified with a PEG end-capping unit, typically an optionally substituted alkyl group, such as -CH3, CH2CH3 or CH2CH2CO2H. A preferred PEG unit has a single polyethylene glycol chain with 2 to 24 -CH2CH2O- subunits covalently linked in series.

[0080] Herein, specific examples of the term "property regulating unit" also include "hydrophilic peptides". The term "hydrophilic peptide" can be connected to the self-degradable linker of the linker unit, or can be connected to the antibody linker, which can be directly connected to the linker unit, or connected to the linker unit via a suitable structural fragment (such as the formula (A) or (A1) fragment defined in the present disclosure). Specifically, "hydrophilic peptide" refers to an organic part comprising a repeating subunit -CO-CR'R"-NR-, wherein one or more amino acids that are identical or different from each other are connected via amide bonds, in a linear, branched or star-shaped configuration, and each hydrophilic peptide generally contains 1-20 amino acids, preferably 4-14 amino acids, and more preferably 6-12 amino acids.

[0081] Herein, the amino acid monomers constituting the hydrophilic peptide may be natural amino acids, such as alanine (Ala), arginine (Arg), aspartic acid (Asp), asparagine (Asn), histidine (His), glycine (Gly), glutamic acid (Glu), glutamine (Gln), phenylalanine (Phe), lysine (Lys), leucine (Leu), serine (Ser), tyrosine (Tyr), threonine (Thr), isoleucine (Ile), proline (Pro), tryptophan (Trp), valine (Val), cysteine ​​(Cys), and methionine (Met), and accordingly, R in the repeating subunit -CO-CR'R"-NR- is H and one of R' and R" is H, and the other corresponds to the corresponding group or fragment in each natural amino acid; preferably polar natural amino acids, such as arginine, serine, threonine, tyrosine, cysteine, aspartic acid, asparagine, glutamic acid, glutamine, lysine, histidine, glycine, and tryptophan.

[0082] In this article, the amino acid monomers constituting the hydrophilic peptide can also be amino acids other than the above-mentioned twenty natural amino acids, such as ornithine (Orn), β-alanine (β-Ala), citrulline (Cit), sarcosine (Sar), and those amino acids in which R, R', and R" in the repeating subunit -CO-CR'R"-NR- are different from the corresponding groups or fragments in natural amino acids, such as R, R', and R" are selected from alkyl, aryl, acyl, amide, ketone, azido, hydroxyl, sulfhydryl, hydrazine, cyano, quaternary ammonium, halogen, hydrazide, alkenyl, alkynyl, ether, thiol, seleno, sulfonic acid, sulfate, borate, phosphono, phosphate, heterocycle, enone, imine, aldehyde, ester, thioacid, hydroxylamine, amino, etc. or any thereof Combinations, or groups comprising alkyl, aryl, acyl, amide, ketone, azido, hydroxyl, sulfhydryl, hydrazine, cyano, quaternary ammonium, halogen, hydrazide, alkenyl, alkynyl, ether, thiol, seleno, sulfonic acid, sulfate, borate, phosphono, phosphate, heterocycle, enone, imine, aldehyde, ester, thioacid, hydroxylamine, amino, etc.; preferably, those amino acids in which R, R' and / or R" comprise a hydrophilic group, for example, R, R', R" are each independently carboxyl, sulfonic acid, sulfate, phosphate, amino, amide, quaternary ammonium, oxygen-containing group, ether, sulfhydryl or hydroxyl, or groups comprising carboxyl, sulfonic acid, sulfate, phosphate, amino, amide, quaternary ammonium, oxygen-containing group, ether, sulfhydryl and / or hydroxyl, for example, alkyl, for example, C 1-6 alkyl.

[0083] For the ADC of the present disclosure, when the linker unit carries a hydrophilic peptide, at least 50% to 100% of the amino acids constituting the hydrophilic peptide are hydrophilic amino acids, such as 80%-100%, for example 60%, 70%, 80%, 90% or 100%, and the amino acids are preferably arginine, serine, threonine, tyrosine, cysteine, aspartic acid, asparagine, glutamic acid, glutamine, lysine, histidine, glycine, tryptophan, ornithine, citrulline (Cit), and sarcosine (Sar).

[0084] For the ADC of the present disclosure, the hydrophilic peptide carried by the linker unit is preferably 4-14 units of polysarcosine, polyarginine, or polyglycine, more preferably 6-12 units of polysarcosine.

[0085] As used herein, the term "drug:antibody ratio" or "DAR" refers to the ratio of the drug moiety (D) conjugated to the Ab moiety described herein to the Ab moiety in an ADC conjugate. In some embodiments described herein, the DAR can be determined by q in Formula I, for example, the DAR can be an integer or non-integer of at least 1, for example, about 1 to 20, for example, about 2-18, 4-16, 5-12, 6-10, 1-10, 1-8, 2-8, 3-8, 2-6, 4-6, 4-8, 6-8, 6-10, for example, about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. DAR can also be calculated as the average DAR of the population of molecules in the product, i.e., the total ratio of the small molecule drug moiety (D) coupled to the Ab moiety described herein to the Ab moiety in the product as measured by a detection method (e.g., by conventional methods such as mass spectrometry, ELISA assay, electrophoresis and / or HPLC), which DAR is referred to herein as the average DAR. In some embodiments, the average DAR value of the conjugates of the present disclosure is about 1 to 20, for example, about 2-18, 4-16, 5-12, 1-10, 1-8, 6-10, 2-8, 3-8, 2-6, 4-6, 4-8, 6-8, 6-10, for example, 1.0-8.0, 2.0-6.0, for example, about 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4 , 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9 or 10.0, and ranges having two of these values ​​as endpoints.

[0086] In this document, the term "halogen" generally refers to fluorine, chlorine, bromine, and iodine, for example, fluorine and chlorine.

[0087] As used herein, the term "alkyl" refers to a straight or branched aliphatic saturated hydrocarbon group having the specified number of carbon atoms. Specifically, the alkyl group may have 1 to 14, 1 to 12, 1 to 10, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2 carbon atoms. Suitable C 1-14 Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, dimethylmethyl, dipropylmethyl, ethylbutylmethyl, diethylmethyl, methylethylmethyl, ethylpropylmethyl, diethylethyl, diethylpropyl, dipropylethyl, etc. Specific alkyl groups have 1 to 7 carbon atoms, e.g., 1 to 6 carbon atoms, 1 to 4 carbon atoms.

[0088] As used herein, the term "-O-alkyl" or "alkoxy" means an alkyl group as defined herein that is attached to the remainder of the molecule through an oxygen atom. Specifically, the -O-alkyl group has 1-10, e.g., 1 to 8, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2 carbon atoms. For example, as used herein, the term "-OC 1-6 The term "alkyl" refers to a straight or branched saturated hydrocarbon group having 1 to 6 carbon atoms which is attached to the rest of the molecule via an oxygen atom, and examples thereof include -O-methyl, -O-ethyl, -O-propyl (including -O-n-propyl and -O-isopropyl), -O-butyl (including -O-n-butyl, -O-isobutyl, -O-sec-butyl or -O-tert-butyl), -O-pentyl (including -O-n-pentyl, -O-isopentyl and -O-neopentyl), -O-n-hexyl, 2-methylpentyl-O-, and the like.

[0089] In this context, the term "C optionally substituted by halogen" 1-6 "Alkyl" refers to the C 1-6 Alkyl, wherein one or more (e.g., 1, 2, 3, 4, or 5) hydrogen atoms are optionally replaced by halogen. It will be understood by those skilled in the art that when there are more than one halogen substituent, the halogens may be the same or different and may be located on the same or different C atoms. 1-6 Examples of "alkyl" are -CH2F, -CHF2, -CF3, -CCl3, -C2F5, -C2Cl5, -CH2CF3, -CH2Cl, -CH2CH2CF3 or -CF(CF3)2, etc.

[0090] In this article, the term "alkenyl" refers to a straight or branched unsaturated hydrocarbon group containing at least one double bond consisting of carbon atoms and hydrogen atoms. Specifically, the alkenyl group has 2-8, such as 2 to 6, 2 to 5, 2 to 4 or 2 to 3 carbon atoms. For example, as used herein, the term "C2-C6 alkenyl" refers to a straight or branched alkenyl group with 2 to 6 carbon atoms, such as vinyl, propenyl, allyl, 1-butenyl, 2-butenyl, 1,3-butadienyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 1,3-pentadienyl, 1,4-pentadienyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 1,4-hexadienyl, etc. The carbon atom connected to the rest of the molecule in the alkenyl group can be saturated or an olefinic carbon atom.

[0091] As used herein, the term "alkynyl" refers to a linear or branched unsaturated hydrocarbon group consisting of carbon atoms and hydrogen atoms and containing at least one triple bond. Specifically, the alkynyl group has 2-8, such as 2 to 6, 2 to 5, 2 to 4 or 2 to 3 carbon atoms. For example, as used herein, the term "C 2-6 "Alkynyl" refers to a straight or branched chain alkynyl group having 2 to 6 carbon atoms, such as ethynyl, propynyl, propargyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-methyl-1-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 5-methyl-2-hexynyl, etc. The carbon atom in the alkynyl group that is connected to the rest of the molecule can be saturated or an acetylenic bond carbon atom.

[0092] As used herein, the term "alkylene" refers to a divalent group obtained by removing two hydrogen atoms from the same or two different carbon atoms of a straight or branched saturated alkane. Specifically, the alkylene group has 1-10 carbon atoms, such as 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2 carbon atoms. For example, as used herein, the term "C 1-6 The term "alkylene" refers to a straight or branched chain alkylene group having 1 to 6 carbon atoms, including but not limited to methylene, ethylene, propylene, butylene, etc.

[0093] In this context, a specific "alkylene" refers to a divalent group obtained by removing two hydrogen atoms from the same carbon atom of a straight-chain or branched saturated alkane, such as the ═C(R d )2 groups, such as structural fragments shown.

[0094] As used herein, the term "alkenylene" refers to a divalent group derived from the removal of two hydrogen atoms from the same or two different carbon atoms of a straight or branched unsaturated olefin containing at least one double bond. Specifically, the alkenylene group has 2-8, such as 2 to 6, 2 to 5, 2 to 4, or 2 to 3 carbon atoms. For example, as used herein, the term "C 2-6 The term "alkenylene" refers to a straight or branched alkenylene group having 2 to 6 carbon atoms, for example, ethenylene, propenylene, allylene, butenylene, pentenylene, and hexenylene.

[0095] As used herein, the term "alkynylene" refers to a divalent group obtained by removing two hydrogen atoms from the same or two different carbon atoms of a straight or branched unsaturated alkyne containing at least one triple bond. Specifically, the alkynylene group has 2-8, such as 2 to 6, 2 to 5, 2 to 4, or 2 to 3 carbon atoms. For example, as used herein, the term "C2-C6 alkynylene" refers to a straight or branched alkynylene group having 2 to 6 carbon atoms, such as ethynylene, propynylene, propargylene, butynylene, pentynylene, and hexynylene.

[0096] As used herein, the term "cycloalkyl" refers to a monocyclic, fused polycyclic, bridged polycyclic, or spirocyclic non-aromatic monovalent hydrocarbon ring structure having the specified number of ring atoms, which may be saturated or unsaturated, e.g., containing one or more double bonds. A cycloalkyl group may contain 3 or more, e.g., 3-18, 3-10, or 3-8 carbon atoms in the ring, e.g., C 3-10 Cycloalkyl, C 3-8 Cycloalkyl, C 3-6 Cycloalkyl, C 5-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

[0097] In this article, the term "heterocycle" or "heterocyclyl" refers to an aromatic or non-aromatic monocyclic, bicyclic or polycyclic ring system with 1-4 heteroatom ring members independently selected from N, O or S. One or more N, C or S atoms in the heterocycle can be oxidized. Preferably, the heterocycle is a 5-10 ring system, which is a monocyclic or fused bicyclic ring. Representative examples include but are not limited to pyrrolidine, azetidine, piperidine, morpholine, tetrahydrofuran, tetrahydropyran, benzofuran, benzothiophene, indole, benzopyrazole, pyrrole, thiophene (thiophene), furan, thiazole, imidazole, pyrazole, pyrimidine, pyridine, pyrazine, pyridazine, isothiazole and isoxazole.

[0098] As used herein, the term "heterocycloalkyl" refers to a monocyclic, fused polycyclic, spirocyclic, or bridged polycyclic non-aromatic saturated ring structure comprising one or more (e.g., 1, 2, 3, or 4) heteroatoms independently selected from O, N, and S and the specified number of ring atoms, or an N-oxide, S-oxide, or S-dioxide thereof. A heterocycloalkyl group may have 3 to 12 ring members (referred to as a 3-12-membered heterocycloalkyl group), for example, 3 to 10 ring members, 3 to 8 ring members, 3 to 7 ring members, 4 to 7 ring members, 4 to 6 ring members, or 5 to 6 ring members. A heterocycloalkyl group typically contains up to 4 (e.g., 1, 2, 3, or 4) heteroatoms, such as a 4-7-membered heterocycloalkyl group containing 1 to 3 heteroatoms selected from N, O, and S. Examples of suitable heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl (e.g., 1-pyrrolidinyl, 2-pyrrolidinyl, and 3-pyrrolidinyl), tetrahydrofuranyl (e.g., 1-tetrahydrofuranyl, 2-tetrahydrofuranyl, and 3-tetrahydrofuranyl), tetrahydrothiophenyl (e.g., 1-tetrahydrothiophenyl, 2-tetrahydrothiophenyl, and 3-tetrahydrothiophenyl), piperidinyl (e.g., 1-piperidinyl), , 2-piperidinyl, 3-piperidinyl and 4-piperidinyl), tetrahydropyranyl (e.g., 4-tetrahydropyranyl), tetrahydrothiopyranyl (e.g., 4-tetrahydrothiopyranyl), morpholinyl (e.g., morpholino), thiomorpholinyl, dioxanyl, piperazinyl or azepanyl, diazepanyl such as 1,4-diazacycloheptyl, 3,6-diaza-bicyclo[3.1.1]heptyl or 3-aza-bicyclo[3.2.1]octyl. The atom in the heterocycloalkyl group that is attached to the rest of the compound can be a carbon atom or a heteroatom, as long as it is chemically feasible.

[0099] In this context, the term "aryl" refers to a monocyclic or polycyclic aromatic hydrocarbon group having 6 to 20, for example 6 to 12, carbon atoms in the ring portion. Preferably, aryl is (C6-C 10 ) aryl. Non-limiting examples include phenyl, biphenyl, naphthyl, or tetrahydronaphthyl, each of which may be optionally substituted with 1 to 4 substituents such as alkyl, trifluoromethyl, cycloalkyl, halogen, hydroxy, alkoxy, acyl, alkyl-C(O)-O-, aryl-O-, heteroaryl-O-, amino, mercapto, alkyl-S-, aryl-S-, nitro, cyano, carboxyl, alkyl-OC(O)-, carbamoyl, alkyl-S(O)-, sulfonyl, sulfonamido, heterocyclyl, and the like.

[0100] As used herein, the term "heteroaryl" refers to a 5-20 membered (e.g., 5-14 membered, 5-8 membered, 5-6 membered) aromatic monocyclic or polycyclic ring system containing 1-4 heteroatoms selected from N, O or S, which may be substituted or unsubstituted. Preferably, the heteroaryl is a 5-10 membered ring system, which is a monocyclic or fused bicyclic ring. Representative heteroaryl groups include 2- or 3-thienyl, 2- or 3-furyl, 2- or 3-pyrrolyl, 2-, 4-, or 5-imidazolyl, 3-, 4-, or 5-pyrazolyl, 2-, 4-, or 5-thiazolyl, 3-, 4-, or 5-isothiazolyl, 2-, 4-, or 5-oxazolyl, 3-, 4-, or 5-isoxazolyl, 3- or 5-1,2,4-triazolyl, 4- or 5-1,2,3-triazolyl, tetrazolyl, 2-, 3-, or 4-pyridyl, 3- or 4-pyridazinyl, 3-, 4-, or 5-pyrazinyl, 2-pyrazinyl, 2-, 4-, or 5-pyrimidinyl.

[0101] As used herein, the term "heteroalkyl" refers to a stable straight or branched chain hydrocarbon that is fully saturated or contains from 1 to 3 degrees of unsaturation, and consists of the indicated number of carbon atoms and from one to ten, preferably from one to three, heteroatoms selected from O, N, Si, and S, wherein the nitrogen and sulfur atoms may be optionally oxidized and the nitrogen heteroatom may be optionally quaternized. The heteroatoms O, N, Si, and S may be placed at any interior position of the heteroalkyl group or at the position at which the heteroalkyl group is attached to the rest of the molecule. Representative examples of heteroalkyl groups include -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2-S(O)-CH3, -NH-CH2-CH2-NH-C(O)-CH2-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=NO-CH3, and -CH=CH-N(CH3)-CH3. Up to two heteroatoms may be consecutive, such as, for example, -CH2-NH-OCH3 and -CH2-O-Si(CH3)3. Typically, a C1 to C4 heteroalkyl or heteroalkylene group has 1 to 4 carbon atoms and 1 or 2 heteroatoms, and a C1 to C3 heteroalkyl or heteroalkylene group has 1 to 3 carbon atoms and 1 or 2 heteroatoms. In some aspects, the heteroalkyl and heteroalkylene groups are saturated.

[0102] Unless otherwise indicated, the term "substituted" as used in the definition of various groups herein means that the corresponding group may be substituted by, for example but not limited to, the following groups as defined herein or conventional in the art: alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, halogen, cyano, nitro, azido, carboxyl, hydroxyl, thiol, amino, mono- or dialkylamino, mono- or dicycloalkylamino, mono- or diarylamino, mono- or diheterocyclylamino, mono- or diheteroarylamino, alkyl- or cycloalkyl- or heterocyclyl- or heteroaryl- or aryl-oxy, alkyl- or cycloalkyl- or heterocyclyl- or heteroaryl- or aryl-thio, Alkyl- or cycloalkyl- or heterocyclyl- or heteroaryl- or aryl-acyl, alkyl- or cycloalkyl- or heterocyclyl- or heteroaryl- or aryl-acylamino, alkyl- or cycloalkyl- or heterocyclyl- or heteroaryl- or aryl-acyloxy, alkyl- or cycloalkyl- or heterocyclyl- or heteroaryl- or aryl-sulfonyl, alkyl- or cycloalkyl- or heterocyclyl- or heteroaryl- or aryl-sulfonyloxy, alkyl- or cycloalkyl- or heterocyclyl- or heteroaryl- or aryl-sulfonylamino, or the above optionally substituted amino-formyl, and each of which is further substituted by remaining optional substituents, wherein each type of group is as defined herein. Examples of substituents include, but are not limited to, one or more groups independently selected from the group consisting of halogen, OH, SH, CN, NH2, NHCH3, N(CH3)2, NO2, N3, C(O)CH3, COOH, C(O)-amino, OCOCH3, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, cyclopropyl, methoxy, ethoxy, propoxy, oxo, trifluoromethyl, difluoromethyl, sulfonylamino, methylsulfonylamino, SO, SO2, phenyl, piperidinyl, piperazinyl, and pyrimidinyl.

[0103] As used herein, the term "substituted" or "substituted" means that one or more (e.g., 1, 2, 3, or 4) hydrogen atoms on the designated atom are replaced with the designated radical, provided that the normal valence of the designated atom in the present context is not exceeded and a stable compound is formed. Combinations of substituents and variables are permissible only if such combinations result in stable compounds.

[0104] As used herein, the term "optionally substituted" means, unless otherwise indicated, that a group may be unsubstituted or substituted with one or more (e.g., 1, 2, 3, 4, or 5 or more, or any range derivatizable therein) of the substituents listed for that group, wherein the substituents may be the same or different. In one embodiment, the optionally substituted group has 1 substituent. In another embodiment, the optionally substituted group has 2 substituents that are the same or different. In another embodiment, the optionally substituted group has 3 substituents that are the same or different. In another embodiment, the optionally substituted group has 4 substituents that are the same or different. In another embodiment, the optionally substituted group has 5 substituents that are the same or different.

[0105] Many of the groups defined herein are optionally substituted, and the listing of substituents given in the definition of a particular group is not intended to limit the substituents defined elsewhere in the specification and claims.

[0106] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that retains the biological effects and properties of the ADC conjugates of the present disclosure and is not biologically or otherwise undesirable. The ADC conjugates of the present disclosure may exist in the form of their pharmaceutically acceptable salts, including acid addition salts and base addition salts. In the present disclosure, a pharmaceutically acceptable, non-toxic acid addition salt refers to a salt formed between the ADC conjugates of the present disclosure and an organic or inorganic acid, including but not limited to hydrochloric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, nitric acid, perchloric acid, acetic acid, oxalic acid, maleic acid, fumaric acid, tartaric acid, benzenesulfonic acid, methanesulfonic acid, salicylic acid, succinic acid, citric acid, lactic acid, propionic acid, benzoic acid, p-toluenesulfonic acid, malic acid, and the like. Pharmaceutically acceptable non-toxic base addition salts refer to salts formed by the ADC conjugates of the present disclosure with organic or inorganic bases, including but not limited to alkali metal salts, such as lithium, sodium or potassium salts; alkaline earth metal salts, such as calcium or magnesium salts; organic base salts, such as ammonium salts formed with organic bases containing N groups.

[0107] As used herein, the term "solvate" refers to an association formed between one or more solvent molecules and the ADC conjugates disclosed herein. Solvents that form solvates include, but are not limited to, water, methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran, N,N-dimethylformamide, dimethyl sulfoxide, and the like. It should be understood that such solvates of the compounds of the invention also include solvates of pharmaceutically acceptable salts of the compounds of the invention.

[0108] As used herein, the term "isotopic variant" refers to a compound wherein one or more atoms constituting the compound are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that may be incorporated into one or more atoms of the disclosed compounds include, for example2 H. 3 H. 13 C. 14 C. 15 N. 17 O. 18 O. 31 P. 32 P. 35 S and 18 F, thereby forming isotopic variations of the compounds of the present disclosure, whether or not they are radioactive, are intended to be included within the scope of the present disclosure. In certain embodiments, the compounds of the present disclosure are provided in an unlabeled form, and in other embodiments, the compounds of the present disclosure are provided in an isotopically labeled form, such as a form labeled with a hydrogen isotope D. In particular, R9, R 10 and R 11 One or more H in the group may be replaced by an isotope D, such as R9 and R 10 are each independently H or D, R 11 Can be substituted with one or more D, especially -C substituted with one or more D 1-6 alkyl.

[0109] As used herein, the term "isomer" refers to any stereoisomer, mixture of enantiomers, including racemates, diastereomeric mixtures, geometric isomers, atropisomers and / or tautomers that may exist in the structure of a compound. The determination and separation methods of the stereochemistry of the isomers are well known to those skilled in the art (SP Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994). The present disclosure encompasses all possible isomeric forms of the compounds defined herein, and pharmaceutically acceptable salts or solvates thereof, unless otherwise indicated. In addition, the compounds of the present disclosure may exist as mixtures of two or more different structural forms in rapid equilibrium (commonly referred to as tautomers). It is to be understood that the scope of this application encompasses all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%).

[0110] Certain compounds of the present disclosure contain at least one (eg, 1, 2, 3, or 4) asymmetric center and can therefore be prepared as (R)- or (S)-stereoisomers, individually or as mixtures thereof.

[0111] The structural formula or structural fragments of the compounds disclosed herein are used Indicates the configuration of a stereocenter, i.e., a chiral center. Accordingly, in the names of the compounds or intermediates provided by the present invention, R or S is used to indicate the configuration of the chiral center. Indicates the racemic form where two configurations of the chiral center exist simultaneously, such as express In some definitions of compounds disclosed herein, axial chirality may also be used to represent compound configurations. These configurations are determined using the Cahn-Ingold-Prelog rules well known to those skilled in the art. The absolute axial chirality configurations of the two example structures shown below are described as follows:

[0112] When the linking axial chiral bond is marked with "*", it means that the compound has a single chiral configuration and is obtained by SFC separation, but the absolute configuration is uncertain, for example, express or

[0113] It should be understood that when a person skilled in the art can determine, based on the structure of the compound shown herein, that the compound exists as a pair of chiral isomers and that the compound can be easily separated based on conventional methods in the art, then the disclosure of the racemate of the compound herein (whether in terms of structural formula or chemical name) should be deemed to have disclosed each isomer of the compound separately.

[0114] In this document, the number of groups attached to each atom in a compound definition, compound formula, or structural fragment depends on the chemical valence of the atom and does not necessarily need to be shown. Generally, only non-hydrogen groups are shown in a group definition, structural formula, or structural fragment. Unshown groups generally represent H. The presence and number of unshown groups can be easily determined by one skilled in the art.

[0115] The structural fragments used in this article The bonds indicated to be cross-linked are the bonds connecting this structural fragment to the rest of the molecule.

[0116] The substituents shown as crossing chemical bonds in the cyclic structure fragments referred to herein are, for example, -(R 12 ) m , refers to the m R 12Substituents may be substituted at any chemically feasible substitution site or sites in the ring, including X where chemically feasible.

[0117] Unless otherwise specified, C in the definition of compounds of the present invention is n-n+m or C n -C m Including various cases from n to n+m carbons, such as C 1-6 Including C1, C2, C3, C4, C5 and C6, and also including any range from n to n+m, such as C 0-6 Including C1, C2, C3, C4, C5, C6, C 0-1 、C 0-2 、C 0-3 、C 0-4 、C 0-5 、C 1-2 、C 1-3 、C 1-4 、C 2-3 etc., C. 1-6 Including C 1-2 、C 1-3 、C 1-4 、C 2-6 、C 3-6 wait.

[0118] As used herein, the word "about" used to modify a numerical value, numerical range, or parameter means that the numerical value or parameter used therewith fluctuates by ±10%, such as ±5%, ±2%, or ±1%. For example, the expression "about 100" as used herein includes 90 and 110 and all values ​​therebetween (e.g., 90.5, 95, 101, 105, 109.95, etc.). For a ratio, the term "about" is used to qualify each digit of the given ratio, for example, a ratio of "about 1:1" means a ratio of (0.9-1.1):(0.9-1.1), and for example, a range of "about nm" or "about n-about m" means 90% n-110% n to 90% m-110% m.

[0119] As used herein, "pharmaceutically acceptable" and "pharmaceutically acceptable" are used interchangeably unless there is any contradiction in the context.

[0120] As used herein, the term "pharmaceutical composition" refers to a composition that is in a form that permits the biological activity of the active ingredient contained therein to be effective, and that contains no additional ingredients that are unacceptably toxic to a subject to which the composition would be administered.

[0121] As used herein, the terms "pharmaceutically acceptable excipient," "pharmaceutically acceptable carrier," and "therapeutically inert excipient" are used interchangeably and refer to any pharmaceutically acceptable ingredient in a pharmaceutical composition that is not therapeutically active and is non-toxic to the subject to which it is administered, such as a disintegrant, binder, filler, solvent, buffer, tonicity agent, stabilizer, antioxidant, surfactant, carrier, diluent, or lubricant used to formulate a drug product.

[0122] As used herein, the term "pharmaceutical combination" refers to a non-fixed combination product or a fixed combination product, including but not limited to a kit and a pharmaceutical composition. The term "non-fixed combination" means that the active ingredients, such as (i) the ADC conjugate of the present invention, and (ii) other therapeutic agents, are administered to a patient simultaneously, without specific time restrictions, or sequentially at the same or different time intervals, as separate entities, wherein such administration provides two or more active agents at prophylactic or therapeutically effective levels in the patient. In some embodiments, the ADC conjugate of the present invention and other therapeutic agents used in the pharmaceutical combination are administered at levels no greater than when they are used alone. The term "fixed combination" means that two or more active agents are administered to a patient simultaneously in the form of a single entity. The dosage and / or time interval of the two or more active agents are preferably selected so that the combined use of the components can produce an effect greater than that achieved by using any one component alone when treating a disease or condition. Each component can be in the form of a separate formulation, which can be the same or different.

[0123] As used herein, the term "combination therapy" refers to the administration of two or more therapeutic agents or treatment modalities, such as radiotherapy or surgery to treat diseases described herein. This administration includes administering these therapeutic agents in a substantially simultaneous manner, such as in a single capsule with a fixed ratio of active ingredients. Alternatively, this administration includes administering each active ingredient in a variety of or separate containers (such as tablets, capsules, powders, and liquids). The powder and / or liquid can be reconstituted or diluted to the desired dose before administration. In addition, this administration also includes using each type of therapeutic agent in a sequential manner at approximately the same time or at different times. In either case, the therapeutic regimen will provide a beneficial effect of the drug combination in treating the illness or condition described herein.

[0124] As used herein, the terms "individual" or "subject" are used interchangeably to refer to mammals. Mammals include, but are not limited to, domesticated animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats). In particular, the subject is a human.

[0125] As used herein, the term "treat," ...

[0126] As used herein, the term "prevent" includes the inhibition of the development or progression of a disease or condition or symptoms of a particular disease or condition. In some embodiments, subjects with a family history of cancer are candidates for a preventative regimen. Generally, in the context of cancer, the term "prevent" refers to the administration of a drug before the development of signs or symptoms of cancer, particularly in a subject at risk for cancer.

[0127] As used herein, the term "effective amount" refers to an amount or dosage of an antibody drug conjugate of the present disclosure, or a composition or combination thereof, that produces the desired effect in a patient in need of treatment or prevention after administration in single or multiple doses to the patient.

[0128] As used herein, the term "therapeutically effective amount" refers to an amount effective to achieve the desired therapeutic outcome at the desired dosage and for the desired period of time. A therapeutically effective amount is also an amount in which any toxic or deleterious effects of the antibody drug conjugates of the present disclosure, or compositions or combinations thereof, are outweighed by the therapeutically beneficial effects. A "therapeutically effective amount" preferably achieves at least about 30%, even more preferably at least about 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or even 100% inhibition of a measurable parameter (e.g., tumor volume) relative to an untreated individual.

[0129] As used herein, the term "prophylactically effective amount" refers to an amount effective to achieve the desired preventive result at the required dosage and for the required period of time. Typically, a prophylactic effective amount will be less than a therapeutically effective amount because a prophylactic dose is used in a subject prior to or at an earlier stage of the disease.

[0130] As used in this specification and the claims that follow, the term "comprising" and variations of the term such as "including" and "comprising" mean "including but not limited to," and are not intended to exclude, for example, other additives, ingredients, integers, or steps. When an element is described as comprising a plurality of ingredients, steps, or conditions, it should be understood that the element may also be described as comprising any combination of the plurality of ingredients, steps, or conditions, or "consisting of" or "consisting essentially of" the plurality or combination of ingredients, steps, or conditions.

[0131] I: Antibody-Drug Conjugates

[0132] In one aspect, the present disclosure provides an antibody-drug conjugate (ADC) having the following formula (X) or a pharmaceutically acceptable salt or solvate thereof: [PL] q -Ab (X)

[0133] in,

[0134] P represents a Ras inhibitor, such as a KRas inhibitor;

[0135] L represents the linker unit that connects P to Ab;

[0136] q represents the number of [PL] units attached to Ab, and is an integer of at least 1 or a non-integer, e.g., q = 1 to 20, e.g., about 1-10, 1-8, 2-8, 3-10, 3-8, 4-10, 4-8, 6-8, or 6-10;

[0137] Ab stands for antibody or antigen-binding fragment.

[0138] In some embodiments of Formula X, q represents an integer selected from 1 to 20, including 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. In some cases, q ranges from 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 1 to 11, 1 to 12, 1 to 13, 1 to 14, 1 to 15, 1 to 16, 1 to 17, 1 to 18, 1 to 19, or 1 to 20, or a range consisting of any two values ​​between 1 and 20, for example, 2 to 10, 2 to 8, 2 to 6, 2 to 4, 3 to 8, 3 to 10, 4 to 6, 4 to 8, 4 to 10, 6 to 8, or 6 to 10. In other embodiments, Formula X describes an ADC in a mixture of ADCs that exhibits a q value ranging from 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 1 to 11, 1 to 12, 1 to 13, 1 to 14, 1 to 15, 1 to 16, 1 to 17, 1 to 18, 1 to 19, or 1 to 20, or a range consisting of any two values ​​between 1 and 20, for example, 2 to 10, 2 to 8, 2 to 6, 2 to 4, 3 to 8, 3 to 10, 4 to 6, 4 to 8, 4 to 10, 6 to 8, or 6 to 10.

[0139] In certain embodiments, Formula X describes the ADCs in a mixture of ADCs such that more than 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of the ADCs in the mixture have a q value of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. In some embodiments, Formula X describes the ADCs in a mixture of ADCs such that more than 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of the ADCs in the mixture have q values ​​in the range of 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 1 to 11, 1 to 12, 1 to 13, 1 to 14, 1 to 15, 1 to 16, 1 to 17, 1 to 18, 1 to 19, or 1 to 20, or a range consisting of any two values ​​between 1 and 20, e.g., 2 to 10, 2 to 8, 2 to 6, 2 to 4, 3 to 8, 3 to 10, 4 to 6, 4 to 8, 4 to 10, 6 to 8, or 6 to 10.

[0140] In other embodiments, Formula X describes an ADC mixture, where q is "q 平均 ”, which represents the average of the q values ​​of the mixture, or the average DAR, i.e., the average number of Linker units (L) attached to a given antibody (Ab) in the mixture. In such an embodiment, q 平均 or average DAR represents an integer or non-integer value from 1 to 20, for example, about 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 1 to 11, 1 to 12, 1 to 13, 1 to 14, 1 to 15, 1 to 16, 1 to 17, 1 to 18, 1 to 19 or 1 to 20, or an integer or decimal within a range consisting of any two values ​​between 1 and 20, for example, about 2 to 10, 2 to 8, 2 to 6, 2 to 4, 3 to 8, 3 to 10, 4 to 6, 4 to 8, 4 to 10, 6 to 8 or 6 to 10.

[0141] In a preferred embodiment, q in Formula (X) of the present disclosure is an integer or non-integer from 1 to 10, or a range consisting of any two values ​​between 1 and 10, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, about 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 2 to 10, 2 to 8, 2 to 6, 2 to 4, 4 to 6, 3 to 8, 3 to 10, 4 to 8, 4-10, 6 to 8, or 6 to 10.

[0142] In some embodiments, q represents an average DAR of about 3. In some embodiments, q represents an average DAR of about 6. In some embodiments, q represents an average DAR of about 8.

[0143] In some embodiments, q represents an average DAR of about 1-10, 1-8, 2-8, 3-10, 3-8, 4-10, 4-8, 6-8, or 6-10.

[0144] It will also be understood by those skilled in the art that the various ADCs described herein may be in the form of salts or solvates, and in some specific embodiments, are pharmaceutically acceptable salts.

[0145] The following describes in detail each component of the ADC conjugates of the present disclosure and the ADC conjugates of the present disclosure composed thereof. Those skilled in the art will appreciate that the ADCs disclosed herein are "modular" in nature, as each comprises the aforementioned modular components Ab, L, and P. Throughout this disclosure, various specific non-limiting embodiments and examples of these modular components are described, and this disclosure encompasses specific combinations of specific embodiments of all modules, as if each specific combination were specifically described separately.

[0146] Ab—antibody or antigen-binding fragment

[0147] In the ADC of the present disclosure, the antibody is an antibody or antigen-binding fragment that specifically binds to the target antigen, and its function is to target and present the KRas mutation inhibitor compound to a specific target cell population. Due to the presence of its targeting component or molecule, the antibody or antigen-binding fragment interacts with the specific target cell population and subsequently releases the free drug within the target cell (intracellular manner) or releases the free drug near the target cell (extracellular manner).

[0148] In one set of embodiments, the antibody or antigen binding fragment is bonded to a linker unit that includes a releasable peptide component. As described above, other connecting components may also be present in the conjugates described herein to provide additional space between Ras, such as a KRas mutation inhibitor compound, and the antibody unit, or to provide the composition with properties that increase solubility. In some of these embodiments, the antibody or antigen binding fragment is bonded to the linker unit via its heteroatoms. The heteroatoms that may be present on the antibody or antigen binding fragment for this bonding include sulfur (in one embodiment, from a sulfhydryl group of a targeting ligand), oxygen (in one embodiment, from a carboxyl or hydroxyl group of a targeting ligand), and optionally substituted nitrogen (in one embodiment, from a primary or secondary amine functional group of a targeting ligand, or in another embodiment, from an optionally substituted amide nitrogen). These heteroatoms may be present on the antibody unit in the natural state of the ligand, such as in naturally occurring antibodies, or may be introduced into the antibody unit by chemical modification or bioengineering.

[0149] The conjugation site on the antibody affects the stability, pharmacokinetic, and pharmacodynamic properties of the ADC. Excessive drug loading can sometimes lead to rapid plasma clearance, while ADCs with a low DAR (drug-antibody ratio) may exhibit weak activity. By selecting the conjugation strategy and the conjugation site on the antibody, the drug loading of the antibody can be controlled while maintaining the structural integrity and homogeneity of the antibody.

[0150] In some embodiments, the antibody or antigen-binding fragment has a sulfhydryl functional group such that it is bonded to the Linker unit via the sulfur atom of the sulfhydryl functional group. In other embodiments, the sulfhydryl group is generated by reducing the interchain disulfides of the antibody. Thus, in some embodiments, the Linker unit is conjugated to a cysteine ​​residue from the reduced interchain disulfide in the antibody. In other embodiments, the sulfhydryl group is chemically introduced into the antibody, for example, by introducing a cysteine ​​residue. Accordingly, in some embodiments, the Linker unit is conjugated to the antibody or antigen-binding fragment via a cysteine ​​residue introduced into the antibody or antigen-binding fragment.

[0151] In other embodiments, the antibody or antigen-binding fragment has one or more lysine residues that are capable of reacting with an activated ester (including but not limited to N-hydroxysuccinimide, pentafluorophenyl, and p-nitrophenyl esters) in the Linker unit, thereby providing an amide bond consisting of a nitrogen atom of the antibody or antigen-binding fragment and the C=O of the Linker unit.

[0152] In other embodiments, the antibody or antigen-binding fragment has one or more lysine residues that can be chemically modified to introduce one or more sulfhydryl groups. In these embodiments, the antibody or antigen-binding fragment is covalently attached to the linker unit via the sulfur atom of the sulfhydryl functional group. Reagents that can be used to modify lysine in this manner include, but are not limited to, N-succinimidyl-S-acetylthioacetate (SATA) and 2-iminothiolane hydrochloride (Traut's reagent).

[0153] In other embodiments, the antibody or antigen-binding fragment has one or more carbohydrate groups that can be modified to provide one or more sulfhydryl functional groups. The chemically modified antibody or antigen-binding fragment in the ADC is bonded to the linker unit via the sulfur atom of the sulfhydryl functional group.

[0154] In other embodiments, the antibody or antigen-binding fragment has one or more carbohydrate groups that can be oxidized to provide an aldehyde (-CHO) functional group. In these embodiments, the corresponding aldehyde interacts with a reactive site on the Linker unit to form a chemical bond between the Linker unit and the Antibody unit.

[0155] In other embodiments, artificial attachment sites are introduced into antibodies to achieve more site-specific conjugation.

[0156] Additional protocols for modifying proteins to attach to linker units or related substances can be found in Coligan et al., Current Protocols in Protein Science, Vol. 2, John Wiley & Sons (2002), which is incorporated herein by reference.

[0157] In some embodiments, the antibody or Fab is able to form a covalent bond between the Linker unit and the antibody or Fab corresponding to the Antibody unit by interacting with the reactive functional group on the Linker unit. The functional group with the ability to interact with the Antibody unit will depend on the properties of the antibody or Fab. In some embodiments, the reactive group is a maleimide. The covalent connection of the antibody or Fab to the Linker unit is achieved by the interaction of the sulfhydryl functional group of the antibody or Fab with the maleimide functional group of the Linker unit to form a sulfur-substituted succinimide. The sulfhydryl functional group can be present in the natural state of the antibody or Fab, such as in a naturally occurring residue, or can be introduced into the antibody or Fab by chemical modification or by bioengineering.

[0158] The antibodies comprising the ADCs of the present disclosure may be polyclonal, monoclonal, genetically engineered, and / or otherwise modified, which are suitable for administration to humans, such as humanized antibodies or fully human antibodies.

[0159] In some embodiments, the Ab unit of the ADC of the present disclosure is a monospecific antibody. In some embodiments, the Ab unit of the ADC of the present disclosure is a multispecific antibody. In some embodiments, the Ab unit, upon binding to an antigen receptor expressed on the surface of a tumor cell, triggers antigen receptor-mediated endocytosis, thereby effectively delivering the anti-tumor drug of the ADC into the tumor cell.

[0160] In some embodiments, the Ab unit of the disclosed ADCs may be a bispecific antibody, a dual variable domain antibody, a multi-chain or single-chain antibody, a single domain antibody, a camelized antibody, a scFv-Fc antibody, a surrogate antibody, or the like. In some embodiments in which the Ab unit is a bispecific antibody, one specificity of the antibody may target a tumor-associated antigen to promote specific binding of the ADC to tumor cells; while the other specificity of the antibody may target a tumor cell surface receptor to further promote ADC internalization and degradation. Examples of such bispecific target combinations include, but are not limited to, the dual targeting combination of HER2 and PRLR on breast cancer cells. In other embodiments, the two specificities of an antibody may each target different tumor-associated antigens to provide a mechanism to combat drug resistance. Examples of such bispecific target combinations include, but are not limited to, the dual targeting combination of EGFR and MET on lung cancer cells. In other embodiments, the two specificities of an antibody may each target different epitopes of the same tumor-associated antigen to increase the antibody's selectivity for cancer cells and / or enhance internalization and trafficking to lysosomes by inducing antigen clustering and cross-linking on the tumor cell surface. Examples of such tumor-associated antigens include, but are not limited to, HER2 on breast cancer cells.

[0161] The antibody portion constituting the ADC of the present disclosure can be in the form of a full-length antibody, which can have or be derived from any antibody isotype, including, for example, IgA, IgD, IgE, IgG, IgM, or IgY. In some embodiments, the antibody constituting the ADC is IgG (e.g., IgG1, IgG2, IgG3, or IgG4). In some embodiments, the antibody constituting the ADC comprises all or part of the constant region of an IgG immunoglobulin.

[0162] The antibody portion constituting the ADC of the present disclosure can be a functionally active fragment, derivative, or analog of an antibody that immunospecifically binds to a target cell (e.g., a cancer cell antigen, a viral antigen, or a microbial antigen). In this regard, "functionally active" means that the fragment, derivative, or analog is able to immunospecifically bind to a target cell.

[0163] Useful antibody fragments include, but are not limited to, F(ab')2 fragments, Fab fragments, Fvs, single-chain antibodies, diabodies, three-chain antibodies, four-chain antibodies, scFv, scFv-FV, or any other molecule with the same specificity as an antibody. The fragments can be obtained by molecular engineering or by chemical or enzymatic treatment of intact antibodies or antibody chains or by recombinant means.

[0164] Useful modified antibody analogs and derivatives include, but are not limited to, derivatives and analogs of antibodies obtained by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, linkage to cellular antibody units or other proteins, etc. Any of a variety of chemical modifications can be performed by known techniques, including but not limited to specific chemical cleavages, acetylation, formylation, metabolic synthesis in the presence of tunicamycin, etc.

[0165] Target antigen and antibody

[0166] The antibody portion of the disclosed ADCs can target any suitable target molecule presented on the surface of target cells, such as a polypeptide, protein, polysaccharide, or lipid molecule. The binding of the antibody to the target molecule should be highly specific to ensure that the ADC specifically binds to the target cell and minimize off-target toxicity. In some embodiments, the binding affinity of the antibody to the target molecule can be selected to be in the nanomolar or subnanomolar range.

[0167] Suitable target antigens can be selected by searching for cell surface proteins that are highly expressed in tumors but lowly expressed or even barely expressed in non-malignant tissues. In some embodiments, such target molecules are membrane antigens expressed on the surface of target tumor cells, such as tumor-specific antigens or tumor-associated antigens, including hematologic tumors and solid tumors, including primary and metastatic tumors. In particularly preferred embodiments, the antibodies or antigen-binding fragments of the ADCs disclosed herein specifically bind to one or more tumor-specific antigens or tumor-associated antigens, or immune cell-associated antigens.

[0168] In some embodiments, the tumor-specific antigen or tumor-associated antigen targeted by the antibody portion of the ADC of the present disclosure is selected from the group consisting of: HER2, Her3, HER1 (ErbB1), HER4 (ErbB4), TROP2, Nectin-4, tissue factor, PD-L1, PD-1, PD-L1 / PD-L2, MET, CLDN18.2, KIT, CTLA-4, RPR1, adrenergic A2 receptor (EphA2), folate receptor (FRa), mesothelin, endothelin receptor, GCPII, IL-13Ra, BCMA, GD2, CLL-1, CA-IX, MUC1, 5T4, AOC3, ALK, AXL, C242, CA-125, CCL11, CCR5, CD2, CD3, CD4, CDS, CD15, CA15-3, CD16, CD18, CD19, CD20, CD21, CD22, CD2 5. CD30, CD32, CD33, CD37, CD38, CD44, CD52, CD56, CD64, CD66e, CD70, CD72, CD74, CD79a, CD79b, CD123, CD138, CD142, CD174, CD276, CDH3, CCD79b, CLDN9 / CLDN6, CA19-9, DPEP3, AGS-16, IGF1R, I GF2R, VEGFR1, VEGFR2, VEGFR3, PDGFR-α, PDGFR-β, EGFR, EGFRvIII, ENPP3, FcRH5, FRα, KAAG1, LIV-1, Mesothelin, cMet, ROR1, SLTRK6, TF, BMPR1B, E16, TOP1, STEAP1, 0772P, MUC16, Napi3b, Sema 5b. PSCAhIg, ETBR, RNF124, prostate cancer-associated gene 1, TrpM4, teratoma-derived growth factor 1, C3DR, FcRH2, NCA, MDP, IL20R-α, Brevican, EphB2R, ASLG659, prostate stem cell antigen precursor, GEDA, BAFF-R, CXCR5, HLA-DOB, P2X5, LY64, FcRH1, IRTA2, TENB2, integrin α5β6, integrin α4β7, FGF2, FGFR1, FGFR2, FGFR3, FGFR4, PSMA, somatostatin receptor, RANK, SLAMF7, ITGB6, CEACAM5, CA9, EG FRvlll, IL2RA, AXL receptor tyrosine kinase, TGF-βR, TNFRSF8, cancer / testis-associated antigen, CLEC14A, GRP78, stem cell-specific antigen, ASG-5, PRR4, GUCY2C, SLC39A6, TPBG, medium tumor-associated antigen CA242, FOLR1, GPNMB, HAVCR1, prostate tumor target Mindin, VTCN1, PTK7 protein tyrosine kinase 7, macrophage-stimulating 1 receptor, TACSTD2, CA6, DLL3, DLL4, EpCAM, FAP, DKK-1, Endoglin, VCAM1, GPC3, DR5, ASCT2, B7H1, B7H3, B7H4.

[0169] In certain embodiments, the tumor-associated antigen and the immune cell antigen are T cell co-inhibitory molecules. In some embodiments, the antibody or its antigen-binding fragment specifically binds to a tumor-associated antigen selected from PD-L1, PD-L2, CD47, CD80, CD86, HVEM, UL144, CD155, CD112, CD113, Galectin-1, Galectin-3, Galectin-9, CD48, LIGHT, BTLA, and CD160. In some embodiments, the tumor-associated antigen is a molecule that binds to a T cell molecule selected from BTLA, Tim-3, PD-1, CTLA-4, TIGIT, CD244, and CD223.

[0170] In some embodiments, the antibody is an anti-PD-L1 antibody, such as atezolizumab, durvalumab, avelumab, or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino acid sequence equivalent thereto.

[0171] In some embodiments, the antibody is an anti-PD-1 antibody, such as nivolumab, pembrolizumab, cemiplimab, anti-mouse PD-1 antibody clone J43, anti-mouse PD-1 antibody clone RMP1-14, mouse anti-PD-1 antibody clone EH12, ANB011, MDX-1106, AMP-514, AMP-224 or Pidilizumab. In some embodiments, the anti-PD-1 antibody is pembrolizumab or nivolumab.

[0172] In some embodiments, the antibody is an anti-CTLA-4 antibody, such as ipilimumab, clone 9H10, tremelimumab, or clone BNI3.

[0173] In some embodiments, the antibody is an anti-CD47 antibody, such as Hu5F9-G4, IBI188, CC-90002, ZL1201, TTI-621, AO-176, an antibody to SGN-CD47M, an ALX148 antigen-binding domain, or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino acid sequence equivalent thereto.

[0174] In other embodiments, the antibody or its antigen-binding fragment specifically binds to a tumor-associated antigen, which is a growth factor receptor (GFR). In certain embodiments, the tumor-associated antigen is an EGFR / ErbB / HER family GFR. In some embodiments, the tumor-associated antigen is selected from EGFR / HER1 (ErbB1), HER2 / c-Neu (ErbB2), Her3 (ErbB3) and Her4 (ErbB4) receptors. In certain embodiments, the tumor-associated antigen is an IGFR family GFR. In some embodiments, the cancer-associated tumor antigen is an IGF1R or IGF2R receptor. In certain embodiments, the tumor-associated antigen is a TGF-βR (TβR) family GFR. In some embodiments, the cancer-associated tumor antigen is a TβR I or TβR II receptor. In certain embodiments, the tumor-associated antigen is a VEGFR family GFR. In some embodiments, the cancer-associated tumor antigen is a VEGFR1, VEGFR2 or VEGFR3 receptor. In certain embodiments, the tumor-associated antigen is a PDGFR family GFR. In some embodiments, the cancer-associated tumor antigen is a PDGFR-α or PDGFR-β receptor. In certain embodiments, the tumor-associated antigen is a FGFR family GFR. In some embodiments, the cancer-associated tumor antigen is a FGFR1, FGFR2, FGFR3, or FGFR4 receptor.

[0175] In some embodiments, the antibody is an anti-EGFR / HER1 (ErbBl) antibody, such as cetuximab, panitumumab, necitumumab, or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino acid sequence equivalent thereto. In some embodiments, the antibody is an anti-HER2 (ErbB2) antibody, such as trastuzumab, pertuzumab, or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino sequence equivalent thereto. In some embodiments, the antibody is an anti-VEGFR2 antibody, such as ramucirumab, or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino sequence equivalent thereto. In some embodiments, the antibody is an anti-PDGFR-α antibody, such as olaratumab, or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino sequence equivalent thereto.

[0176] In other embodiments, the antibody or its antigen-binding fragment specifically binds to a lymphoma-associated antigen. In certain embodiments, the lymphoma-associated antigen is CD20, CD30, CD19 / CD3, CD22, or CD33. In some embodiments, the antibody is an anti-CD20 antibody, such as rituximab, ibritumomab, ofatumumab, obinutuzumab, or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino acid sequence equivalent thereto. In some embodiments, the antibody is an anti-CD30 antibody, such as brentuximab, or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino acid sequence equivalent thereto. In some embodiments, the antibody is an anti-CD19 / CD3 antibody, such as blinatumomab, or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino acid sequence equivalent thereto. In some embodiments, the antibody is an anti-CD22 antibody, such as inotuzumab or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino acid sequence equivalent thereto. In some embodiments, the antibody is an anti-CD33 antibody, such as gemtuzumab or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino acid sequence equivalent thereto.

[0177] In other embodiments, the antibody or its antigen-binding fragment specifically binds to a myeloma-associated antigen. In certain embodiments, the myeloma-associated antigen is SLAMF7 or CD38. In some embodiments, the antibody is an anti-SLAMF7 antibody, such as elotuzumab or its antigen-binding fragment, or an antibody or its antigen-binding fragment having an amino acid sequence equivalent thereto. In some embodiments, the antibody is an anti-CD38 antibody, such as daratumumab or its antigen-binding fragment, or an antibody or its antigen-binding fragment having an amino acid sequence equivalent thereto.

[0178] In other embodiments, the antibody or antigen-binding fragment thereof specifically binds to a germ cell tumor-associated antigen. In certain embodiments, the germ cell tumor-associated antigen is GD2. In some embodiments, the antibody is an anti-GD2 antibody, such as dinutuximab or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino acid sequence equivalent thereto.

[0179] In other embodiments, the antibody or antigen-binding fragment thereof specifically binds to a RANK ligand. In some embodiments, the antibody is an anti-RANK ligand antibody, such as denosumab or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino acid sequence equivalent thereto.

[0180] In other embodiments, the antibody or its antigen-binding fragment specifically binds to TROP2. In some embodiments, the antibody is an anti-TROP2 antibody, such as certolizumab and datopotamab, or an antigen-binding fragment thereof or an antibody or antigen-binding fragment thereof having an amino acid sequence equivalent thereto.

[0181] In other embodiments, the antibody or antigen-binding fragment thereof specifically binds to Claudin 18.2. In some embodiments, the antibody is an anti-Claudin 18.2 antibody, such as Zolbetuximab, Osemitamab (TST001), CMG901, ASKB589, ZL-1211, or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino acid sequence equivalent thereto.

[0182] In other embodiments, the antibody or its antigen-binding fragment specifically binds to Met (also known as c-Met, or hepatocyte growth factor receptor (HGFR)). A pathogenic mutation in the Met gene encodes an abnormal Met receptor, which transmits abnormal signals and causes multiple effects, including cell growth, survival, invasion, metastasis, angiogenesis, etc. Tumors involved in Met include non-small cell lung cancer, colorectal cancer, gastric cancer, esophageal cancer, glioma, etc. In some embodiments, the antibody is an anti-Met antibody, such as onartuzumab or its antigen-binding fragment, or an antibody or antigen-binding fragment having an amino acid sequence equivalent thereto.

[0183] In other embodiments, the antibody or its antigen-binding fragment specifically binds to the adrenergic receptor Eph, particularly the EphA2 receptor. EphA2 is not only a biomarker of malignant characteristics, but also an active participant in malignant progression. It has been shown to play an important role in the regulation of cancer occurrence and tumor progression. Human EphA2 is abundantly expressed in prostate cancer, lung cancer, esophageal cancer, colorectal cancer, cervical cancer, ovarian cancer, breast cancer, and skin cancer, which is associated with poor prognosis, increased metastatic potential, and reduced survival in tumor patients. In some embodiments, the antibody is an anti-EphA2 antibody or its antigen-binding fragment, or an antibody or its antigen-binding fragment having an amino acid sequence equivalent thereto.

[0184] In other embodiments, the antibody or antigen-binding fragment thereof specifically binds to nectin-4. In some embodiments, the antibody is an anti-nectin-4 antibody, such as enfortumab or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino acid sequence equivalent thereto.

[0185] In other embodiments, the antibody or antigen-binding fragment thereof specifically binds to both EGFR and Met (also known as c-Met). In some embodiments, the antibody is an anti-EGFR and Met bispecific antibody, such as Amivantamab or an antigen-binding fragment thereof, or an antibody or antigen-binding fragment thereof having an amino acid sequence equivalent thereto.

[0186] In some embodiments, the tumor-associated antigen to which the antibody or antigen-binding fragment thereof specifically binds is selected from AXL, B7H1, B7H3, B7H4, BCMA, CD16, CD19, CD22, CD25, CD30, CD32, CD33, CD44, CD64, CD70, CD74, CD79, CD138, CD142, CD276, CDH3, CEACAM5, Claudin 18.2, CLDN9 / CLDN6, DPEP3, EGFR, ENPP3, EphA, FcRH5, FOLR1, FRα, GCPII, HER2, HER3, KAAG1, KIT, LIV-1, Mesothelin, c-Met, MUC1, Nectin-4, PD-L1, PD-L1 / PD-L2, PSMA, ROR1, RPR1, TF, TOP1, TROP2, etc. The corresponding antibodies are commercially available or can be prepared by techniques known in the art.

[0187] In preferred embodiments, the antibodies are those that bind to antigens that are preferentially expressed or overexpressed in cancer cells, such as HER2 (ErbB2), PD-1, PD-L1, EGFR, TROP2, Claudin 18.2, EphA-2, Nectin-4, and Met, more preferably HER2 (ErbB2), EGFR, TROP2, Claudin 18.2, Nectin-4, or a combination of EGFR and Met.

[0188] Targeting the tumor-associated antigens mentioned above, the ADC compounds disclosed herein can treat tumors associated with the expression of the above-mentioned antigens. The specific list of tumors can be known or determined by those skilled in the art based on existing technologies.

[0189] Antibodies immunospecific for tumor-associated antigens can be obtained commercially or produced by any method known to those skilled in the art, such as recombinant expression techniques. Nucleotide sequences encoding antibodies immunospecific for cancer cell antigens can be obtained, for example, from the GenBank database or similar databases, literature publications, or by conventional cloning and sequencing.

[0190] HER2-targeting Ab unit

[0191] In a particularly preferred embodiment, the antibody conjugates provided herein include antibodies or antigen-binding fragments (anti-HER2 antibodies) that specifically bind to human HER2, i.e., the antibody portion specifically targets the tumor-associated antigen HER2. Therefore, in some aspects, the disclosure provides an antibody-drug conjugate (ADC) comprising an antibody or antigen-binding fragment thereof that specifically binds to HER2 as the Ab unit of the ADC.

[0192] HER2 is a receptor tyrosine protein kinase, ErbB2, which is overexpressed or amplified in approximately 20-30% of breast cancers. Increased activation of HER2 triggers multiple downstream pathways, leading to abnormal proliferation of cancer cells (Treish I, Schwartz R, Lindley C: Pharmacology and therapeutic use of trastuzumab in breast cancer. Am J Health Syst Pharm. 2000 Nov 15; 57(22): 2063-76; quiz 2077-9). HER2 is also overexpressed in many other types of cancer, such as gastric cancer, esophageal cancer, colon cancer, rectal cancer, breast cancer, ovarian cancer, cervical cancer, uterine cancer, endometrial cancer, bladder cancer, pancreatic cancer, lung cancer, prostate cancer, osteosarcoma, neuroblastoma, or head and neck cancer.

[0193] In some embodiments, the antibody or antibody fragment thereof (e.g., antigen-binding fragment) that specifically binds to human HER2 can be selected from trastuzumab, pertuzumab, margetuximab or HT-19 or an antibody fragment thereof or a site-specific mutant thereof, or other anti-human HER2 antibodies that recognize the same epitope or compete for binding to human HER2.

[0194] Trastuzumab (trade name Herceptin or Herclon) is a humanized monoclonal antibody against human epidermal growth factor 2 used to treat HER2-positive breast cancer, gastrointestinal cancer, and gastric cancer. It binds to the membrane-proximal portion of the extracellular domain of the HER2 receptor. The amino acid sequences of its heavy and light chain variable regions are described in U.S. Patent 5,821,337. It interacts with three loop regions formed by human HER2 residues 557–561, 570–573, and 593–603 (Cho et al., Nature 421:756-760, 2003). It may interfere with HER2 signaling by preventing HER2 receptor dimerization, promoting HER2 receptor endocytosis, and inhibiting the shedding of the extracellular domain (Hudis CA, N Engl J Med. 2007; 357(1):39-51). Another important mechanism of action of anti-HER2 antibodies is the mediation of antibody-dependent cellular cytotoxicity (ADCC). In ADCC, anti-HER2 antibodies bind to tumor cells and then recruit immune cells, such as macrophages, through Fcε receptor (FcεR) interactions. Trastuzumab was approved by the U.S. FDA in September 1998 for the treatment of patients with metastatic breast cancer.

[0195] Pertuzumab (also known as 2C4, Omnitarg, Perjeta) is a humanized monoclonal antibody that binds to the extracellular domain of the HER2 receptor and inhibits the dimerization of HER2 with other HER receptors. The amino acid sequences of its heavy and light chains are described in U.S. Patent 7,560,111. Pertuzumab interacts primarily with residues within the 245–333 region of human HER2, particularly residues His 245, Val 286, Ser 288, Leu 295, His 296, or Lys 311 (Franklin et al., Cancer Cell 5:317–328, 2004). Pertuzumab has been shown to be more effective than trastuzumab in disrupting HER1-HER2 and HER3-HER2 complex formation in breast and prostate cancer cell lines (Agus et al., J Clin Oncol. 2005; 23(11): 2534-43. Epub Feb 7, 2005). For efficacy, pertuzumab does not require antibody-dependent cellular cytotoxicity because it does not require an intact Fc region for activity (Agus et al., J Clin Oncol. 2005; 23(11): 2534-43. Epub Feb 7, 2005). Pertuzumab is approved by the U.S. FDA for use in combination with trastuzumab and docetaxel for the treatment of patients with HER2-positive metastatic breast cancer.

[0196] Margetuximab (also known as MGAH22) is another anti-HER2 monoclonal antibody (see http: / / www.macrogenics.com / products-margetuximab.html). The Fc region of margetuximab has been optimized to increase its binding to activating FcεRs. Margetuximab is currently in clinical trials for the treatment of patients with recurrent or refractory advanced breast cancer whose tumors express HER2 2+ by immunohistochemistry and lack evidence of HER2 gene amplification by FISH.

[0197] HT-19 is another anti-HER2 monoclonal antibody that binds to an epitope in human HER2 that is distinct from that of trastuzumab or pertuzumab and has been shown to inhibit HER2 signaling comparable to trastuzumab and, in combination with trastuzumab and pertuzumab, promotes HER2 degradation (Bergstrom DA et al., Cancer Res. 2015;75:LB-231).

[0198] Exemplary antibodies targeting HER2 that can be used in the ADC of the present disclosure can be antibodies or antigen-binding fragments comprising all six CDR sequences of an antibody selected from the group consisting of: trastuzumab (Herceptin, Genentech, US6,054,297); ATCC Accession Nos. PTA-10355, PTA-10356, PTA-10357, PTA 10358 (US20100119511); ATCC Accession No. CRL-10463 (Genentech); ATCC Accession Nos. HB-12215, HB-12216, CRL 10463, HB-12697; Pertuzumab (Genentech, US20110117097); ATCC Accession Nos. HB-12215, HB-12216, CRL 10463, HB-12697; 10463, HB-12698 (US20090202546); ATCC accession number HB-12215, HB-12216 (US20060088523); ATCC accession number (7C2) HB-12215, (7F3) HB-12216, (4D5) CRL-10463, (2C4) HB-12697 (US20060018899); TrasGEX (Glycotope: http: / / www.glycotope.conn / pipeline). The present disclosure also contemplates anti-HER2 antibodies disclosed in the following documents: US201 10177095, US20100119511, US20110117097, US20090285837, US20090202546, US20060088523, US20060018899, US2011 / 0159014, US20090187007, US20110217305.

[0199] >Trastuzumab light chain amino acid sequence (SEQ ID NO: 1)

[0200] >Trastuzumab heavy chain amino acid sequence (SEQ ID NO: 2)

[0201] >Trastuzumab light chain LCDR1 sequence (SEQ ID NO: 3)

[0202] >Trastuzumab light chain LCDR2 sequence (SEQ ID NO: 4)

[0203] >Trastuzumab light chain LCDR3 sequence (SEQ ID NO: 5)

[0204] >Trastuzumab heavy chain HCDR1 sequence (SEQ ID NO: 6)

[0205] >Trastuzumab heavy chain HCDR2 sequence (SEQ ID NO: 7)

[0206] >Trastuzumab heavy chain HCDR3 sequence (SEQ ID NO: 8)

[0207] >Trastuzumab light chain variable region VL (SEQ ID NO: 9)

[0208] >Trastuzumab heavy chain variable region VH (SEQ ID NO: 10)

[0209] In one embodiment, the antibody portion used in the ADC of the present disclosure comprises all six CDR sequences of trastuzumab. In another embodiment, the antibody portion used in the ADC of the present disclosure comprises the heavy chain variable region sequence and the light chain variable region sequence of trastuzumab. In yet another embodiment, the antibody portion used in the ADC of the present disclosure comprises the heavy chain sequence and the light chain sequence of trastuzumab.

[0210] In some embodiments, the Ab unit of the ADC of the present disclosure comprises three CDRs of the heavy chain variable region (VH) sequence of SEQ ID NO: 10 and three CDRs of the light chain variable region (VL) sequence of SEQ ID NO: 9, and preferably, wherein the CDRs are defined according to Kabat or IMGT or a combination thereof.

[0211] In some embodiments, the Ab unit of the disclosed ADC comprises three heavy chain complementarity determining regions (HCDRs) and three light chain complementarity determining regions (LCDRs), wherein:

[0212] According to the Kabat definition, HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 6, HCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 7, HCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 8, LCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 3, LCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 4, and LCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 5.

[0213] In one embodiment, the Ab unit of the ADC of the present disclosure comprises a heavy chain variable region, wherein the heavy chain variable region comprises: the amino acid sequence shown in SEQ ID NO: 10, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0214] In one embodiment, the Ab unit of the ADC of the present disclosure comprises a light chain variable region, wherein the light chain variable region comprises: the amino acid sequence shown in SEQ ID NO: 9, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0215] In some preferred embodiments, the Ab unit of the ADC of the present disclosure comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 10, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 9.

[0216] In some embodiments, the Ab unit of the ADC of the present disclosure preferably further comprises a heavy chain constant region and / or a light chain constant region of an antibody. Preferably, the heavy chain constant region is a heavy chain constant region derived from a human immunoglobulin. Preferably, the light chain constant region is a light chain constant region derived from a human immunoglobulin. In some aspects, the heavy chain constant region contained in the Ab unit can be any isotype or subtype, such as a heavy chain constant region of an IgG1, IgG2, IgG3 or IgG4 isotype, and preferably an IgG1, IgG2 or IgG4 heavy chain constant region, especially a human IgG1 heavy chain constant region. In further aspects, the light chain constant region contained in the Ab unit can be a kappa light chain constant region or a lambda light chain constant region, especially a human kappa light chain constant region.

[0217] In some embodiments, the Ab unit of the ADC of the present disclosure comprises a human IgG1 heavy chain constant region. Preferably, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 11, or an amino acid sequence comprising at least one, two, or three, but not more than 20, 10, or 5 amino acid changes relative to the amino acid sequence of SEQ ID NO: 11, or a sequence having at least 95-99% identity with the amino acid sequence of SEQ ID NO: 11.

[0218] An exemplary amino acid sequence of a human IgG1 heavy chain constant region (SEQ ID NO: 11)

[0219] In some embodiments, the Ab unit of the ADC of the present disclosure comprises a human kappa light chain constant region. Preferably, the light chain constant region comprises the amino acid sequence of SEQ ID NO: 12, or an amino acid sequence comprising at least one, two, or three, but not more than 20, 10, or 5 amino acid changes relative to the amino acid sequence of SEQ ID NO: 12, or a sequence having at least 95-99% identity with the amino acid sequence of SEQ ID NO: 12.

[0220] An exemplary amino acid sequence of a human kappa light chain constant region (SEQ ID NO: 12)

[0221] In some embodiments, the Ab unit of the ADC of the present disclosure is a full-length antibody comprising a heavy chain constant region and a light chain constant region. In some embodiments, the Ab unit is a tetrameric structure formed by two light chains and two heavy chains. In further embodiments, the Ab unit is an IgG antibody, particularly an IgG1 antibody.

[0222] In some preferred embodiments, the Ab unit of the ADC of the present disclosure comprises a heavy chain and a light chain, wherein: the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 2, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or consists thereof. In other preferred embodiments, the Ab unit of the ADC of the present disclosure comprises a heavy chain and a light chain, wherein: the light chain comprises the amino acid sequence shown in SEQ ID NO: 1, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0223] In some more preferred embodiments, the Ab unit of the ADC of the present disclosure comprises:

[0224] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 2, and

[0225] (b) A light chain comprising the amino acid sequence of SEQ ID NO: 1.

[0226] Ab unit targeting TROP2

[0227] In a particularly preferred embodiment, the antibody conjugate provided herein includes an antibody or antigen-binding fragment (anti-TROP2 antibody) that specifically binds to human TROP2, i.e., the antibody portion specifically targets the tumor-associated antigen TROP2. Therefore, in some aspects, the present disclosure provides an antibody-drug conjugate (ADC) comprising an antibody or antigen-binding fragment thereof that specifically binds to TROP2 as the Ab unit of the ADC.

[0228] Trophoblast cell surface antigen 2 (TROP2), also known as tumor-associated calcium signal transducer 2 (TACSTD2), is a type I transmembrane cell surface glycoprotein. The human TROP2 sequence is available at UniProtKB accession number P09758. TROP2 has been shown to be overexpressed in many solid tumors, including, but not limited to, various human epithelial cancers, such as cervical, endometrial, breast, urothelial, lung, gastric, prostate, colorectal, and pancreatic cancers. Furthermore, TROP2 can play a role in tumor cell proliferation, invasion, migration, apoptosis, and therapeutic resistance by binding to or interacting with a variety of molecules. These characteristics make TROP2 an attractive pan-cancer target for cancer therapy. Ying Wen et al., A literature review of the promising future of TROP2: a potential drug therapy target, Ann Transl Med. 2022Dec; 10(24):1403, doi:10.21037 / atm-22-5976.

[0229] Antibodies targeting human TROP2 that can be used in the ADCs of the present disclosure can be prepared using antibody preparation processes known in the art. For example, anti-TROP2 antibodies can be obtained by immunizing an animal with human TROP2 (UniProtKB Accession No. P09758) or a polypeptide comprising the amino acid sequence of the extracellular domain of TROP2, harvesting antibodies from the immunized animal, and purifying and preferably humanizing the antibodies. In addition, fully human anti-human TROP2 antibodies can be obtained using yeast display libraries expressing human immunoglobulin sequences or transgenic animals.

[0230] In some embodiments, the antibody or antibody fragment thereof (e.g., antigen-binding fragment) that specifically binds to human TROP2 can be selected from Sacituzumab, Datopotamab, or an antibody fragment thereof, or other anti-human TROP2 antibodies that recognize the same epitope or compete for binding to human TROP2.

[0231] Sacituzumab, also known as certolizumab pegol, is a humanized version of the murine monoclonal antibody RS7, developed by Immunomedics. It is a humanized IgG1κ monoclonal antibody targeting TROP2. This antibody can directly bind to cancer cells expressing TROP2 and trigger its internalization. The sequence of sacituzumab can be found in US10179171B2.

[0232] Datopotamab is an IgG1 anti-TROP2 antibody. It was generated by humanizing a mouse mAb that specifically binds to human TROP2. The sequence of datopotamab can be found in Daisuke Okajima et al., Datopotamab Deruxtecan, a Novel TROP2-directed Antibody–drug Conjugate, Demonstrates Potent Antitumor Activity by Efficient Drug Delivery to Tumor Cells, Mol Cancer Ther (2021) 20(12): 2329–2340.

[0233] Exemplary antibodies targeting human TROP2 that can be used for the ADC of the present disclosure can be antibodies or antigen-binding fragments comprising all 6 CDR sequences of an antibody selected from the group consisting of Sacituzumab and Datopotamab (preferably, comprising heavy chain variable region and light chain variable region sequences of an antibody selected from the group consisting of Sacituzumab and Datopotamab). The present disclosure also contemplates anti-human TROP2 antibodies disclosed in the following documents: WO2010089782A1; US2021 / 0393792 A1; WO2008 / 144891, WO2011 / 145744, WO2011 / 155579, WO2013 / 077458, WO2003 / 074566, WO2011 / 068845, WO2013 / 068946, US 2023 / 0270870A1.

[0234] >Sacituzumab light chain amino acid sequence (SEQ ID NO: 13)

[0235] >Sacituzumab heavy chain amino acid sequence (SEQ ID NO: 14)

[0236] >Sacituzumab light chain LCDR1 sequence (SEQ ID NO: 15)

[0237] >Sacituzumab light chain LCDR2 sequence (SEQ ID NO: 16)

[0238] >Sacituzumab light chain LCDR3 sequence (SEQ ID NO: 17)

[0239] >Sacituzumab heavy chain HCDR1 sequence (SEQ ID NO: 18)

[0240] >Sacituzumab heavy chain HCDR2 sequence (SEQ ID NO: 19)

[0241] >Sacituzumab heavy chain HCDR3 sequence (SEQ ID NO: 20)

[0242] >Sacituzumab light chain variable region VL (SEQ ID NO: 21)

[0243] >Sacituzumab heavy chain variable region VH (SEQ ID NO: 22)

[0244] In one embodiment, the antibody portion used in the ADC of the present disclosure comprises all 6 CDR sequences of Sacituzumab or Datopotamab. In another embodiment, the antibody portion used in the ADC of the present disclosure comprises the heavy chain variable region sequence and the light chain variable region sequence of Sacituzumab or Datopotamab. In yet another embodiment, the antibody portion used in the ADC of the present disclosure comprises the heavy chain sequence and the light chain sequence of Sacituzumab or Datopotamab.

[0245] In some embodiments, the Ab unit of the ADC of the present disclosure comprises three CDRs of the heavy chain variable region (VH) sequence of SEQ ID NO: 22 and three CDRs of the light chain variable region (VL) sequence of SEQ ID NO: 21, and preferably, wherein the CDRs are defined according to Kabat or IMGT or a combination thereof.

[0246] In some embodiments, the Ab unit of the disclosed ADC comprises three heavy chain complementarity determining regions (HCDRs) and three light chain complementarity determining regions (LCDRs), wherein:

[0247] According to the Kabat definition, HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 18, HCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 19, HCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 20, LCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 15, LCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 16, and LCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 17.

[0248] In one embodiment, the Ab unit of the ADC of the present disclosure comprises a heavy chain variable region, wherein the heavy chain variable region comprises: the amino acid sequence shown in SEQ ID NO: 22, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0249] In one embodiment, the Ab unit of the ADC of the present disclosure comprises a light chain variable region, wherein the light chain variable region comprises: the amino acid sequence shown in SEQ ID NO: 21, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0250] In some preferred embodiments, the Ab unit of the ADC of the present disclosure comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 22, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 21.

[0251] In some embodiments, the Ab unit of the ADC of the present disclosure preferably further comprises a heavy chain constant region and / or a light chain constant region of an antibody. Preferably, the heavy chain constant region is a heavy chain constant region derived from a human immunoglobulin. Preferably, the light chain constant region is a light chain constant region derived from a human immunoglobulin. In some aspects, the heavy chain constant region contained in the Ab unit can be any isotype or subtype, such as a heavy chain constant region of an IgG1, IgG2, IgG3 or IgG4 isotype, and preferably an IgG1, IgG2 or IgG4 heavy chain constant region, especially a human IgG1 heavy chain constant region. In further aspects, the light chain constant region contained in the Ab unit can be a kappa light chain constant region or a lambda light chain constant region, especially a human kappa light chain constant region.

[0252] In some embodiments, the Ab unit of the ADC of the present disclosure comprises a human IgG1 heavy chain constant region. Preferably, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 23, or an amino acid sequence comprising at least one, two, or three, but not more than 20, 10, or 5 amino acid changes relative to the amino acid sequence of SEQ ID NO: 23, or a sequence having at least 95-99% identity with the amino acid sequence of SEQ ID NO: 23.

[0253] An exemplary amino acid sequence of a human IgG1 heavy chain constant region (SEQ ID NO: 23)

[0254] In some embodiments, the Ab unit of the ADC of the present disclosure comprises a human kappa light chain constant region. Preferably, the light chain constant region comprises the amino acid sequence of SEQ ID NO: 12, or an amino acid sequence comprising at least one, two, or three, but not more than 20, 10, or 5 amino acid changes relative to the amino acid sequence of SEQ ID NO: 12, or a sequence having at least 95-99% identity with the amino acid sequence of SEQ ID NO: 12.

[0255] In some embodiments, the Ab unit of the ADC of the present disclosure is a full-length antibody comprising a heavy chain constant region and a light chain constant region. In some embodiments, the Ab unit has a tetrameric structure formed by two light chains and two heavy chains. In further embodiments, the Ab unit is an IgG antibody, particularly an IgG1 antibody.

[0256] In some preferred embodiments, the Ab unit of the ADC of the present disclosure comprises a heavy chain and a light chain, wherein: the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 14, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or consists thereof. In other preferred embodiments, the Ab unit of the ADC of the present disclosure comprises a heavy chain and a light chain, wherein: the light chain comprises the amino acid sequence shown in SEQ ID NO: 13, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0257] In some more preferred embodiments, the Ab unit of the ADC of the present disclosure comprises:

[0258] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 14, and

[0259] (b) A light chain comprising the amino acid sequence of SEQ ID NO: 13.

[0260] Cancers that can be treated with the disclosed ADCs targeting TROP2 include, but are not limited to, adenocarcinoma, squamous cell carcinoma, non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), colorectal cancer, gastric adenocarcinoma, esophageal cancer, hepatocellular carcinoma, ovarian epithelial cancer, breast cancer, metastatic breast cancer, triple-negative breast cancer (TNBC), prostate cancer, hormone-refractory prostate cancer, pancreatic ductal adenocarcinoma, head and neck cancer, renal cell carcinoma, bladder tumors, cervical cancer, endometrial cancer, uterine cancer, follicular thyroid cancer, and glioblastoma multiforme.

[0261] Ab unit targeting Claudin18.2

[0262] In a particularly preferred embodiment, the antibody conjugates provided herein include antibodies or antigen-binding fragments that specifically bind to human Claudin18.2 (anti-Claudin18.2 antibodies), i.e., the antibody portion specifically targets the tumor-associated antigen Claudin18.2. Therefore, in some aspects, the present disclosure provides antibody-drug conjugates (ADCs) comprising an antibody or antigen-binding fragment thereof that specifically binds to Claudin18.2 as the Ab unit of the ADC.

[0263] Claudin 18.2 (also abbreviated as CLDN18.2) is a tight junction membrane protein belonging to the claudin family. Claudin 18.2 sequences from humans and various mammals can be found in UniProtKB. For example, the human claudin 18.2 sequence can be found in UniProtKB accession number P56856-2. Expression of this protein in healthy tissues is primarily restricted to differentiated gastric mucosal epithelial cells, but it exhibits abnormal overexpression in a range of malignant tumors, particularly those of the digestive system. Therefore, claudin 18.2 has been proposed as a promising target for the development of antibody-based ADC cancer therapeutics. Daisuke Kyuno et al., Claudin-18.2 as a therapeutic target in cancers: cumulative findings from basic research and clinical trials, Tissue Barriers. 2022; 10(1):1967080.doi:10.1080 / 21688370.2021.1967080; Jinxia Chen, Targeting CLDN18.2 in cancers of the gastrointestinal tract: New drugs and new indications, Front Oncol. 2023;13:1132319, doi:10.3389 / fonc.2023.1132319.

[0264] Antibodies targeting human Claudin18.2 that can be used in the ADCs of the present disclosure can be prepared using antibody preparation processes known in the art. For example, anti-Claudin18.2 antibodies can be obtained by immunizing animals with human Claudin18.2 (UniProtKB Accession No. P56856-2) or a polypeptide comprising the amino acid sequence of the extracellular domain of Claudin18.2, harvesting antibodies from the immunized animals, and purifying and preferably humanizing them. In addition, fully human anti-human Claudin18.2 antibodies can be obtained using yeast display libraries or transgenic animals expressing human immunoglobulin sequences.

[0265] In some embodiments, the antibody or antibody fragment thereof (e.g., antigen-binding fragment) that specifically binds to human Claudin18.2 can be selected from Zolbetuximab, Osemitamab (TST001), CMG901, ASKB589, ZL-1211, or an antibody fragment thereof, or other anti-human Claudin18.2 antibodies that recognize the same epitope or compete for binding to human Claudin18.2.

[0266] Zolbetuximab (also known as GC-182, IMAB-362, IMAB362, claudiximab) is an IgG1 antibody derived from a murine monoclonal antibody that has been chimerized to display human IgG1 constant regions for clinical use. This antibody can directly bind to cancer cells expressing CLDN18.2 and trigger its internalization. The sequence of zolbetuximab can be found in WO2007059997 and WO2016 / 165762.

[0267] Osemitamab (TST001) is a high-affinity humanized anti-Claudin18.2 antibody. It exhibits enhanced antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), and demonstrates potent anti-tumor activity in tumor xenograft models. The sequence of this antibody is available from Inxight Drugs and INN (code 11927).

[0268] CMG901, ASKB589, and ZL-1211 are exemplary antibodies against human CLDN18.2 currently in clinical trials (see DOI: 10.1200 / JCO.2023.41.4_suppl.352; DOI: 10.1200 / JCO.2023.41.4_suppl.397; DOI: 10.1200 / JCO.2023.41.16_suppl.2537).

[0269] Exemplary antibodies targeting human CLDN18.2 that can be used in the ADC of the present disclosure can be antibodies or antigen-binding fragments comprising all six CDR sequences of an antibody selected from the group consisting of Zolbetuximab, Osemitamab (TST001), and CMG901 (preferably, comprising heavy chain variable region and light chain variable region sequences of an antibody selected from the group consisting of).

[0270] The present disclosure also contemplates anti-human Claudin18.2 antibodies disclosed in the following documents: WO2007059997A1, CN107667118A, WO2016 / 166122, US11555070B2, WO2020 / 135674, WO2018 / 006882, CN109762067, WO2019 / 242505, WO2020 / 038404, WO2020 / 043044, WO2020 / 063988, WO2020 / 082209, WO2020 / 018852, WO2020 / 023679, WO2020 / 135674, WO2020 / 135201, WO2020 / 139956, WO2020 / 025792, WO2020160560, CN111808194, and WO2020200196.

[0271] >Zolbetuximab light chain amino acid sequence (SEQ ID NO: 24)

[0272] >Zolbetuximab heavy chain amino acid sequence (SEQ ID NO: 25)

[0273] >Zolbetuximab light chain LCDR1 sequence (SEQ ID NO: 26)

[0274] >Zolbetuximab light chain LCDR2 sequence (SEQ ID NO: 27)

[0275] >Zolbetuximab light chain LCDR3 sequence (SEQ ID NO: 28)

[0276] >Zolbetuximab heavy chain HCDR1 sequence (SEQ ID NO: 29)

[0277] >Zolbetuximab heavy chain HCDR2 sequence (SEQ ID NO: 30)

[0278] >Zolbetuximab heavy chain HCDR3 sequence (SEQ ID NO: 31)

[0279] >Zolbetuximab light chain variable region VL (SEQ ID NO: 32)

[0280] >Zolbetuximab heavy chain variable region VH (SEQ ID NO: 33)

[0281] In one embodiment, the antibody portion used in the ADC of the present disclosure comprises all six CDR sequences of Zolbetuximab. In another embodiment, the antibody portion used in the ADC of the present disclosure comprises the heavy chain variable region sequence and the light chain variable region sequence of Zolbetuximab. In yet another embodiment, the antibody portion used in the ADC of the present disclosure comprises the heavy chain sequence and the light chain sequence of Zolbetuximab.

[0282] In some embodiments, the Ab unit of the ADC of the present disclosure comprises three CDRs of the heavy chain variable region (VH) sequence of SEQ ID NO: 33 and three CDRs of the light chain variable region (VL) sequence of SEQ ID NO: 32, and preferably, wherein the CDRs are defined according to Kabat or IMGT or a combination thereof.

[0283] In some embodiments, the Ab unit of the disclosed ADC comprises three heavy chain complementarity determining regions (HCDRs) and three light chain complementarity determining regions (LCDRs), wherein:

[0284] According to the Kabat definition, HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 29, HCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 30, HCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 31, LCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 26, LCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 27, and LCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 28.

[0285] In one embodiment, the Ab unit of the ADC of the present disclosure comprises a heavy chain variable region, wherein the heavy chain variable region comprises: the amino acid sequence shown in SEQ ID NO: 33, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0286] In one embodiment, the Ab unit of the ADC of the present disclosure comprises a light chain variable region, wherein the light chain variable region comprises: the amino acid sequence shown in SEQ ID NO: 32, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0287] In some preferred embodiments, the Ab unit of the ADC of the present disclosure comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 33, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 32.

[0288] In some embodiments, the Ab unit of the ADC of the present disclosure preferably further comprises a heavy chain constant region and / or a light chain constant region of an antibody. Preferably, the heavy chain constant region is a heavy chain constant region derived from a human immunoglobulin. Preferably, the light chain constant region is a light chain constant region derived from a human immunoglobulin. In some aspects, the heavy chain constant region contained in the Ab unit can be any isotype or subtype, such as a heavy chain constant region of an IgG1, IgG2, IgG3 or IgG4 isotype, and preferably an IgG1, IgG2 or IgG4 heavy chain constant region, especially a human IgG1 heavy chain constant region. In further aspects, the light chain constant region contained in the Ab unit can be a kappa light chain constant region or a lambda light chain constant region, especially a human kappa light chain constant region.

[0289] In some embodiments, the Ab unit of the ADC of the present disclosure comprises a human IgG1 heavy chain constant region. Preferably, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 34, or an amino acid sequence comprising at least one, two, or three, but not more than 20, 10, or 5 amino acid changes relative to the amino acid sequence of SEQ ID NO: 34, or a sequence having at least 95-99% identity with the amino acid sequence of SEQ ID NO: 34.

[0290] An exemplary amino acid sequence of a human IgG1 heavy chain constant region (SEQ ID NO: 34)

[0291] In some embodiments, the Ab unit of the ADC of the present disclosure comprises a human kappa light chain constant region. Preferably, the light chain constant region comprises the amino acid sequence of SEQ ID NO: 12, or an amino acid sequence comprising at least one, two, or three, but not more than 20, 10, or 5 amino acid changes relative to the amino acid sequence of SEQ ID NO: 12, or a sequence having at least 95-99% identity with the amino acid sequence of SEQ ID NO: 12.

[0292] In some embodiments, the Ab unit of the ADC of the present disclosure is a full-length antibody comprising a heavy chain constant region and a light chain constant region. In some embodiments, the Ab unit is a tetrameric structure formed by two light chains and two heavy chains. In further embodiments, the Ab unit is an IgG antibody, particularly an IgG1 antibody.

[0293] In some preferred embodiments, the Ab unit of the ADC of the present disclosure comprises a heavy chain and a light chain, wherein: the heavy chain comprises the amino acid sequence of SEQ ID NO: 25, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or consists thereof. In other preferred embodiments, the Ab unit of the ADC of the present disclosure comprises a heavy chain and a light chain, wherein: the light chain comprises the amino acid sequence of SEQ ID NO: 24, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0294] In some more preferred embodiments, the Ab unit of the ADC of the present disclosure comprises:

[0295] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 25, and

[0296] (b) a light chain comprising the amino acid sequence of SEQ ID NO: 24.

[0297] Cancers that can be treated with the disclosed ADCs targeting Claudin18.2 include, but are not limited to, various digestive tract cancers, such as gastric cancer (GC), gastroesophageal junction (GEJ) cancer, esophageal cancer, and pancreatic cancer.

[0298] EGFR-targeting Ab unit

[0299] In a particularly preferred embodiment, the antibody conjugates provided herein include antibodies or antigen-binding fragments (anti-EGFR antibodies) that specifically bind to human EGFR, i.e., the antibody portion specifically targets the tumor-associated antigen EGFR. Therefore, in some aspects, the present invention provides antibody-drug conjugates (ADCs) comprising an antibody or antigen-binding fragment thereof that specifically binds to EGFR as the Ab unit of the ADC.

[0300] Epidermal growth factor receptor, also abbreviated herein as EGFR, is a member of the epidermal growth factor receptor (HER) family and is encoded by the c-erbB proto-oncogene (also known as HER-1 or Erb-B1). EGFR sequences for humans and various mammals can be found in UniProtKB. For example, the human EGFR sequence is found in UniProtKB accession number P00533. EGFR is overexpressed in many solid tumors, including lung cancer, head and neck cancer, breast cancer, kidney cancer, gastric cancer, colon cancer, pancreatic cancer, ovarian cancer, prostate cancer, and bladder cancer; and EGFR can induce tumor proliferation by homodimerization.

[0301] EGFR has been proposed as a promising target for antibody-based cancer therapeutics. To date, five EGFR-targeting monoclonal antibodies have been approved for clinical cancer treatment, namely cetuximab ( ), panitumumab ( ), nimotuzumab (nimotuzumab, BIOMAB- ), necitumuma and amivantamab (amivantamab-vmjw; In addition, three EGFR-based ADC drugs are in clinical trials. However, no EGFR-based ADC has yet been approved for treatment. See Jinfeng Yu et al., Antibody-Drug Conjugates Targeting the Human Epidermal Growth Factor Receptor Family in Cancers, Front Mol Biosci. 2022;9:847835, doi:10.3389 / fmolb.2022.847835.

[0302] A variety of anti-EGFR monoclonal antibodies have been developed for binding to the extracellular domain of the receptor to block the interaction between the receptor and the ligand or its dimerization. These antibodies are all suitable for the present invention. In addition, antibodies targeting human EGFR that can be used for the ADC of the present invention can also be prepared using antibody preparation processes known in the art. For example, anti-EGFR antibodies can be obtained in the following manner: immunizing animals with human EGFR (UniProtKB accession number P00533) or a polypeptide comprising the amino acid sequence of the EGFR extracellular domain, harvesting antibodies from the immunized animals, and purifying and preferably humanizing them. In addition, yeast display libraries or transgenic animals expressing human immunoglobulin sequences can be used to obtain antibodies against human EGFR with complete human sequences.

[0303] The antibody targeting human EGFR used in the ADC of the present invention can be a monospecific antibody that binds to EGFR. In some cases, the antibody targeting human EGFR used in the ADC of the present invention can also be a multispecific antibody, especially a bispecific antibody, such as a bispecific antibody targeting EGFR and MET; or a bispecific antibody targeting MUC1 and EGFR.

[0304] In some embodiments, the anti-human EGFR antibody or antibody fragment thereof (eg, antigen-binding fragment) used in the ADC of the present invention can be selected from cetuximab ( ), panitumumab ( ), nimotuzumab (nimotuzumab, ), necitumuma Depatuxizumab (ABT-806), NECITUMUMAB, IZALONTAMAB, BAFISONTAMAB, Petosemtamab, PIMURUTAMAB, FUTUXIMAB, MODOTUXIMAB (Zatuximab) or their antibody fragments, or other anti-human EGFR antibodies that recognize the same epitope or compete for binding to human EGFR. The sequences of the above antibodies can be found in Inxight Drugs and International Nonproprietary Names (INN) codes 7906, 8499, 8545, 9083, 11030, 10263, 9083, 12022, 11851, 11136, 11309, 9612, and 9613.

[0305] Cetuximab (cetuximab, ) is a recombinant chimeric human / mouse IgG1 monoclonal antibody. This antibody binds to the extracellular domain of the inactive EGFR with a much higher affinity than the endogenous ligand. This antibody competitively blocks ligand-receptor binding, inhibiting the ligand's agonistic effect on the receptor. It also promotes EGFR internalization and downregulates EGFR expression on the cell membrane. Furthermore, this antibody can activate antibody-dependent cell-mediated cytotoxicity (ADCC), resulting in further cell-killing effects. The sequence of cetuximab can be found in WO2007092453 and INN code 7906.

[0306] Panitumumab (panitumumab, ) is a recombinant humanized IgG2 monoclonal antibody. Panitumumab specifically binds to EGFR on tumor cells and competitively inhibits EGFR ligand binding. Preclinical studies have shown that panitumumab binding to EGFR prevents ligand-induced receptor autophosphorylation and activation of receptor-associated kinases, thereby inhibiting cell growth, inducing apoptosis, reducing the production of proinflammatory cytokines and angiogenic factors, and inducing EGFR internalization. The sequence of panitumumab can be found at INN code 8499.

[0307] Exemplary antibodies targeting human EGFR that can be used in the ADC of the present invention can be antibodies or antigen-binding fragments comprising all six CDR sequences of an antibody selected from the group consisting of cetuximab, panitumumab, nimotuzumab, necitumuma, depatuxizumab, NECITUMUMAB, IZALONTAMAB, BAFISONTAMAB, Petosemtamab, PIMURUTAMAB, FUTUXIMAB and MODOTUXIMAB.

[0308] The present invention also contemplates the anti-human EGFR antibodies disclosed in the following documents: WO2023040941A1, WO2022271722A1, WO2022159576A1, WO2022128716A1, WO2022105878A1, WO2021247798A1, WO2022104697A1, WO2021066869A1, WO2020233534A1, WO2020130125A1, WO2019046858A1, WO2019046859A1, WO2019035630A2, WO2019035630A3, WO2018098035A1, WO201721423 3A1, WO2017214282A1, WO2017214301A1, WO2017161206A1, WO2017139623A1, WO2017136581A1, WO2017076492A1, WO2017060322A3, WO2017025458A1, WO2017008169A1, WO2016065456A1, WO2015143382A1, WO2014143765A1, WO2014143765A8, WO2014152199A1, WO2014094355A1, WO2012143495A2, and WO2012143495A3.

[0309] >Cetuximab light chain amino acid sequence (SEQ ID NO: 35)

[0310] >Cetuximab heavy chain amino acid sequence (SEQ ID NO: 36)

[0311] >Cetuximab light chain LCDR1 sequence (SEQ ID NO: 37)

[0312] >Cetuximab light chain LCDR2 sequence (SEQ ID NO: 38)

[0313] >Cetuximab light chain LCDR3 sequence (SEQ ID NO: 39)

[0314] >Cetuximab heavy chain HCDR1 sequence (SEQ ID NO: 40)

[0315] >Cetuximab heavy chain HCDR2 sequence (SEQ ID NO:41)

[0316] >Cetuximab heavy chain HCDR3 sequence (SEQ ID NO: 42)

[0317] >Cetuximab light chain variable region VL (SEQ ID NO: 43)

[0318] >Cetuximab heavy chain variable region VH (SEQ ID NO: 44)

[0319] In one embodiment, the antibody portion used in the ADC of the present invention comprises all six CDR sequences of Cetuximab. In another embodiment, the antibody portion used in the ADC of the present invention comprises the heavy chain variable region sequence and the light chain variable region sequence of Cetuximab. In yet another embodiment, the antibody portion used in the ADC of the present invention comprises the heavy chain sequence and the light chain sequence of Cetuximab.

[0320] In some embodiments, the Ab unit of the ADC of the present invention comprises three CDRs of the heavy chain variable region (VH) sequence of SEQ ID NO: 44 and three CDRs of the light chain variable region (VL) sequence of SEQ ID NO: 43, and preferably, wherein the CDRs are defined according to Kabat or IMGT or a combination thereof.

[0321] In some embodiments, the Ab unit of the ADC of the present invention comprises three heavy chain complementarity determining regions (HCDRs) and three light chain complementarity determining regions (LCDRs), wherein:

[0322] According to the Kabat definition, HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO:40, HCDR2 comprises or consists of the amino acid sequence of SEQ ID NO:41, HCDR3 comprises or consists of the amino acid sequence of SEQ ID NO:42, LCDR1 comprises or consists of the amino acid sequence of SEQ ID NO:37, LCDR2 comprises or consists of the amino acid sequence of SEQ ID NO:38, and LCDR3 comprises or consists of the amino acid sequence of SEQ ID NO:39.

[0323] In one embodiment, the Ab unit of the ADC of the present invention comprises a heavy chain variable region, wherein the heavy chain variable region comprises: an amino acid sequence as shown in SEQ ID NO: 44, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0324] In one embodiment, the Ab unit of the ADC of the present invention comprises a light chain variable region, wherein the light chain variable region comprises: an amino acid sequence as shown in SEQ ID NO: 43, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0325] In some preferred embodiments, the Ab unit of the ADC of the present invention comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 44, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 43.

[0326] In some embodiments, the Ab unit of the ADC of the present invention preferably further comprises a heavy chain constant region and / or a light chain constant region of an antibody. Preferably, the heavy chain constant region is a heavy chain constant region derived from a human immunoglobulin. Preferably, the light chain constant region is a light chain constant region derived from a human immunoglobulin. In some aspects, the heavy chain constant region contained in the Ab unit can be of any isotype or subtype, such as a heavy chain constant region of the IgG1, IgG2, IgG3, or IgG4 isotype, and preferably an IgG1, IgG2, or IgG4 heavy chain constant region, particularly a human IgG1 heavy chain constant region. In further aspects, the light chain constant region contained in the Ab unit can be a kappa light chain constant region or a lambda light chain constant region, particularly a human kappa light chain constant region.

[0327] In some embodiments, the Ab unit of the ADC of the present invention comprises a human IgG1 heavy chain constant region. Preferably, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 34, or an amino acid sequence comprising at least one, two, or three, but not more than 20, 10, or 5 amino acid changes relative to the amino acid sequence of SEQ ID NO: 34, or a sequence having at least 95-99% identity with the amino acid sequence of SEQ ID NO: 34.

[0328] In some embodiments, the Ab unit of the ADC of the present invention comprises a human kappa light chain constant region. Preferably, the light chain constant region comprises the amino acid sequence of SEQ ID NO: 12, or an amino acid sequence comprising at least one, two, or three, but not more than 20, 10, or 5 amino acid changes relative to the amino acid sequence of SEQ ID NO: 12, or a sequence having at least 95-99% identity with the amino acid sequence of SEQ ID NO: 12.

[0329] In some embodiments, the Ab unit of the ADC of the present invention is a full-length antibody comprising a heavy chain constant region and a light chain constant region. In some embodiments, the Ab unit has a tetrameric structure formed by two light chains and two heavy chains. In further embodiments, the Ab unit is an IgG antibody, particularly an IgG1 antibody.

[0330] In some preferred embodiments, the Ab unit of the ADC of the present invention comprises a heavy chain and a light chain, wherein: the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 36, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or consists thereof. In other preferred embodiments, the Ab unit of the ADC of the present invention comprises a heavy chain and a light chain, wherein: the light chain comprises the amino acid sequence shown in SEQ ID NO: 35, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0331] In some more preferred embodiments, the Ab unit of the ADC of the present invention comprises:

[0332] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 36, and

[0333] (b) A light chain comprising the amino acid sequence of SEQ ID NO: 35.

[0334] Cancers that can be treated with the ADCs of the present disclosure that target EGFR include, but are not limited to, various primary and metastatic solid tumors, such as lung cancer (e.g., lung adenocarcinoma, lung squamous cell carcinoma, and non-small cell lung cancer), head and neck cancer (e.g., head and neck squamous cell carcinoma), nasopharyngeal cancer, esophageal cancer, biliary tract cancer, colon cancer, colorectal cancer, pancreatic cancer, gastric cancer, and glioblastoma.

[0335] Ab units targeting EGFR and MET

[0336] In a particularly preferred embodiment, the antibody conjugates provided herein include antibodies or antigen-binding fragments (anti-EGFR / MET antibodies) that specifically bind to human EGFR and MET (also known as c-Met), i.e., the antibody portion specifically targets the tumor-associated antigens EGFR and MET. Therefore, in some aspects, the present disclosure provides antibody-drug conjugates (ADCs) comprising an antibody or antigen-binding fragment thereof that specifically binds to EGFR and MET as the Ab unit of the ADC.

[0337] Drug resistance is a major challenge facing targeted cancer therapy. For example, MET amplification or protein overexpression has been identified as an important mechanism of clinical resistance to EGFR inhibitors. Similarly, emerging evidence suggests that activation of the EGFR pathway may lead to resistance to c-MET targeted inhibitors. Benedettini, E., et al., Met activation in non-small cell lung cancer is 2010 associated with de novo resistance to EGFR inhibitors and the development of brain metastasis. The American journal of pathology, 2010. 177(1): p. 415-423. Bertotti, 2015 Bertotti, A. and F. Sassi, Molecular Pathways: Sensitivity and Resistance to Anti-EGFR Antibodies. Clinical Cancer Research, 2015. In addition, co-expression of cMet and EGFR has been observed in a variety of cancers, including non-small cell lung cancer, pancreatic cancer, colorectal cancer, head and neck squamous cell carcinoma, breast cancer, and esophageal-gastric cancer. Therefore, drug molecules based on multispecific antibodies (such as bispecific antibodies) targeting EGFR and c-MET have been proposed as a promising treatment option for EGFR / cMET single-positive and double-positive tumors. Examples of EGFR / cMET dual-targeting antibodies include, but are not limited to, Amivantamab, AZD9592, Pamvatamig (MCLA-129), Bafisontamab (EMB01), and LY-3164530, which have been approved or are in clinical development.

[0338] In some embodiments, antibodies or antibody fragments thereof (e.g., antigen-binding fragments) that specifically bind to human EGFR and MET are selected from the group consisting of: Amivantamab, AZD9592, Pamvatamig (MCLA-129), Bafisontamab (EMB01), and LY-3164530, or antibody fragments or site-specific mutants thereof, or other anti-human EGFR and MET antibodies that recognize the same epitope or competitively bind to human EGFR and MET. These antibodies can promote endocytosis at relevant targets and enter tumor cells for degradation, and therefore are suitable drug delivery vehicles.

[0339] Amivantamab, also known as JNJ-61186372, is an anti-EGFR-MET bispecific antibody. The antibody has an EGFR antibody arm consisting of a heavy chain and a light chain, and a MET antibody arm consisting of a heavy chain and a light chain, with a 1+1 asymmetric IgG-like structure. The antibody binds to EGFR and MET, acts as an EGFR antagonist and MET inhibitor, and induces the internalization of EGFR and MET on the cell surface. Amivantamab is suitable for the treatment of various cancers with EGFR exon 20 insertion mutations, especially non-small cell lung cancer. Patients with non-small cell lung cancer often develop resistance to drugs that target EGFR and MET alone. The development of Amivantamab reduces the possibility of resistance development by targeting both EGFR and MET, and has shown superior efficacy to single EGFR inhibitors and single MET inhibitors in animal models and clinical trials. The FDA has approved this antibody for the treatment of patients with metastatic non-small cell lung cancer (NSCLC) harboring EGFR exon 20 insertion mutations that have progressed after platinum-based chemotherapy. The antibody sequence of Amivantamab can be found in US20230174677A1 and CAS No. 2171511-58-1.

[0340] AZD9592 is a drug molecule based on a MET-EGFR dual-target antibody currently in Phase 1 clinical research and is suitable for colorectal cancer, head and neck tumors, and non-small cell lung cancer. The bispecific antibody RAA22 / B09-57 targeting EGFR and c-Met involved in this drug molecule simultaneously targets both EGFR and c-Met and is a 1+1 asymmetric bispecific antibody. The antibody works by inhibiting the hepatocyte growth factor receptor MET and the epidermal growth factor receptor EGFR. The antibody sequence of RAA22 / B09-57 can be found in US2023 / 0183358A1.

[0341] Pamvatamig, also known as MCLA-129, is a 1+1 asymmetric bispecific antibody. This bispecific antibody utilizes a DEKK mutation to prevent heavy chain mispairing and a common light chain to prevent light chain mispairing. Functionally, MCLA-129 not only inhibits the phosphorylation of cMet and EGFR in tumors but also reverses tumor resistance to erlotinib. This bispecific antibody is suitable for targeting advanced malignant solid tumors, colorectal cancer, esophageal squamous cell carcinoma, metastatic non-small cell lung cancer, head and neck squamous cell carcinoma, gastric cancer, advanced non-small cell lung cancer, and non-small cell lung cancer with EGFR mutations. The antibody sequence of Pamvatamig is disclosed under US Pat. No. 11773170B2 and CAS No. 2750004-05-6.

[0342] Bafisontamab (EMB01) is a EMB-01 is a bispecific antibody targeting EGFR and c-Met, constructed using a bispecific antibody platform. Structurally, EMB-01 has a symmetrical 2+2 bispecific antibody structure. EMB-01 induced significantly stronger anti-tumor activity in PDX and CDX tumor models than either anti-EGFR or anti-c-Met monoclonal antibodies alone, and demonstrated significant anti-tumor activity in anti-EGFR monoclonal antibody-resistant tumor models. The antibody sequence of EMB-01 is disclosed in WO2017136820 (as molecule FIT013a) and under INN code 11851.

[0343] LY-3164530 is a c-MET / EGFR-targeting bispecific antibody. It fuses the scFv (cetuximab) to the N-terminus of the heavy chain of emibetuzumab (LY2875358), forming a 2+2 symmetrical bispecific antibody. This antibody is of the IgG4 subtype and, therefore, lacks effector functions such as ADCC. In a mouse tumor model, this bispecific antibody demonstrated modest inhibition of tumor cell growth. The antibody sequence of LY-3164530 is publicly available under CAS 2069210-01-9.

[0344] Exemplary antibodies targeting EGFR and MET that can be used in the ADC of the present disclosure can be antibodies or antigen-binding fragments comprising all six CDR sequences that specifically bind to EGFR and all six CDR sequences that specifically bind to MET of an antibody selected from the group consisting of Amivantamab, AZD9592, Pamvatamig (MCLA-129), Bafisontamab (EMB01), and LY-3164530. The present disclosure also contemplates the anti-EGFR and MET antibodies disclosed in WO2019031965A1, WO2022104236A2, WO2023069888A1, WO2023122588A2, WO2023172133A1, WO2023172134A1, WO2024002938A1, and WO2024153168A2.

[0345] >Anti-EGFR heavy chain amino acid sequence of ivantuzumab (SEQ ID NO: 45)

[0346] >Anti-EGFR light chain amino acid sequence of ivantuzumab (SEQ ID NO: 46)

[0347] >Anti-MET heavy chain amino acid sequence of ivantuzumab (SEQ ID NO: 47)

[0348] >Anti-MET light chain amino acid sequence of ivantuzumab (SEQ ID NO: 48)

[0349] >Anti-EGFR heavy chain HCDR1 amino acid sequence of ivantuzumab (SEQ ID NO: 49)

[0350] >Anti-EGFR heavy chain HCDR2 amino acid sequence of ivantuzumab (SEQ ID NO: 50)

[0351] >Anti-EGFR heavy chain HCDR3 amino acid sequence of ivantuzumab (SEQ ID NO: 51)

[0352] >Anti-EGFR light chain LCDR1 amino acid sequence of ivantuzumab (SEQ ID NO: 52)

[0353] >Anti-EGFR light chain LCDR2 amino acid sequence of ivantuzumab (SEQ ID NO: 53)

[0354] >Anti-EGFR light chain LCDR3 amino acid sequence of ivantuzumab (SEQ ID NO: 54)

[0355] >Anti-MET heavy chain HCDR1 amino acid sequence of ivantuzumab (SEQ ID NO: 55)

[0356] >Anti-MET heavy chain HCDR2 amino acid sequence of ivantuzumab (SEQ ID NO: 56)

[0357] >Anti-MET heavy chain HCDR3 amino acid sequence of ivanovumab (SEQ ID NO: 57)

[0358] >Anti-MET light chain LCDR1 amino acid sequence of ivantuzumab (SEQ ID NO: 58)

[0359] >Anti-MET light chain LCDR2 amino acid sequence of ivantuzumab (SEQ ID NO: 59)

[0360] >Anti-MET light chain LCDR3 amino acid sequence of ivantuzumab (SEQ ID NO: 60)

[0361] >Anti-EGFR heavy chain variable region VH amino acid sequence of ivantuzumab (SEQ ID NO: 61)

[0362] >Anti-EGFR light chain variable region VL amino acid sequence of ivantuzumab (SEQ ID NO: 62)

[0363] >Anti-MET heavy chain variable region VH amino acid sequence of ivantuzumab (SEQ ID NO: 63)

[0364] >Anti-MET light chain variable region VL amino acid sequence of ivantuzumab (SEQ ID NO: 64)

[0365] In one embodiment, the antibody portion used in the ADC of the present invention comprises all 6 CDR sequences in the anti-EGFR arm of entuzumab and all 6 CDR sequences in the anti-MET arm. In another embodiment, the antibody portion used in the ADC of the present invention comprises the heavy chain variable region sequence and the light chain variable region sequence of the anti-EGFR arm of entuzumab and the heavy chain variable region sequence and the light chain variable region sequence of the anti-MET arm. In yet another embodiment, the antibody portion used in the ADC of the present invention comprises the anti-EGFR heavy chain sequence and the light chain sequence of entuzumab and the anti-MET heavy chain sequence and the light chain sequence.

[0366] In some embodiments, the Ab unit of the ADC of the present invention comprises a first set of complementarity determining regions that bind to EGFR and a second set of complementarity determining regions that bind to MET, wherein the first set of complementarity determining regions comprises three CDRs of the heavy chain variable region (VH) sequence of SEQ ID NO: 61 and three CDRs of the light chain variable region (VL) sequence of SEQ ID NO: 62, and the second set of complementarity determining regions comprises three CDRs of the heavy chain variable region (VH) sequence of SEQ ID NO: 63 and three CDRs of the light chain variable region (VL) sequence of SEQ ID NO: 64. Preferably, the CDRs are defined according to Kabat or IMGT or a combination thereof.

[0367] In some embodiments, the Ab unit of the ADC of the present invention comprises a first set of complementarity determining regions that bind to EGFR and a second set of complementarity determining regions that bind to MET, wherein:

[0368] The first set of complementarity determining regions comprises three heavy chain complementarity determining regions (HCDRs) and three light chain complementarity determining regions (LCDRs), wherein:

[0369] According to the Kabat definition, HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO:49, HCDR2 comprises or consists of the amino acid sequence of SEQ ID NO:50, HCDR3 comprises or consists of the amino acid sequence of SEQ ID NO:51, LCDR1 comprises or consists of the amino acid sequence of SEQ ID NO:52, LCDR2 comprises or consists of the amino acid sequence of SEQ ID NO:53, and LCDR3 comprises or consists of the amino acid sequence of SEQ ID NO:54;

[0370] The second set of complementarity determining regions comprises three heavy chain complementarity determining regions (HCDRs) and three light chain complementarity determining regions (LCDRs), wherein:

[0371] According to the Kabat definition, HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 55, HCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 56, HCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 57, LCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 58, LCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 59, and LCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 60.

[0372] In one embodiment, the Ab unit of the ADC of the present invention comprises a first heavy chain variable region and a first light chain variable region that bind to EGFR and a second heavy chain variable region and a second light chain variable region that bind to MET. In some embodiments, the first heavy chain variable region comprises: the amino acid sequence set forth in SEQ ID NO: 61, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto. In some embodiments, the first light chain variable region comprises: the amino acid sequence set forth in SEQ ID NO: 62, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto. In some embodiments, the second heavy chain variable region comprises: the amino acid sequence set forth in SEQ ID NO: 63, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto. In some embodiments, the second light chain variable region comprises: the amino acid sequence shown in SEQ ID NO:64, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0373] In some embodiments, the Ab unit of the ADC of the present invention comprises a first heavy chain variable region and a first light chain variable region that bind to EGFR and a second heavy chain variable region and a second light chain variable region that bind to MET, wherein:

[0374] The first heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 61, and the first light chain variable region comprises the amino acid sequence of SEQ ID NO: 62; and

[0375] The second heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 63, and the second light chain variable region comprises the amino acid sequence shown in SEQ ID NO: 64.

[0376] In some embodiments, the Ab unit of the ADC of the present disclosure preferably further comprises a heavy chain constant region and / or a light chain constant region of an antibody. Preferably, the heavy chain constant region is a heavy chain constant region derived from a human immunoglobulin. Preferably, the light chain constant region is a light chain constant region derived from a human immunoglobulin. In some aspects, the heavy chain constant region contained in the Ab unit can be any isotype or subtype, such as a heavy chain constant region of an IgG1, IgG2, IgG3 or IgG4 isotype, and preferably an IgG1, IgG2 or IgG4 heavy chain constant region, especially a human IgG1 heavy chain constant region. In further aspects, the light chain constant region contained in the Ab unit can be a kappa light chain constant region or a lambda light chain constant region, especially a human kappa light chain constant region.

[0377] In some embodiments, the Ab unit of the ADC of the present disclosure comprises a human IgG1 heavy chain constant region. Preferably, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 65 or 66, or an amino acid sequence comprising at least one, two, or three, but not more than 20, 10, or 5 amino acid changes relative to the amino acid sequence of SEQ ID NO: 65 or 66, or a sequence having at least 95-99% identity with the amino acid sequence of SEQ ID NO: 65 or 66.

[0378] An exemplary amino acid sequence of a human IgG1 heavy chain constant region (SEQ ID NO: 65)

[0379] An exemplary amino acid sequence of a human IgG1 heavy chain constant region (SEQ ID NO: 66)

[0380] In some embodiments, the Ab unit of the ADC of the present disclosure comprises a human kappa light chain constant region. Preferably, the light chain constant region comprises the amino acid sequence of SEQ ID NO: 12, or an amino acid sequence comprising at least one, two, or three, but not more than 20, 10, or 5 amino acid changes relative to the amino acid sequence of SEQ ID NO: 12, or a sequence having at least 95-99% identity with the amino acid sequence of SEQ ID NO: 12.

[0381] In some embodiments, the Ab unit of the ADC of the present disclosure comprises or consists of a first heavy chain, a first light chain, a second heavy chain, and a second light chain, wherein from N-terminus to C-terminus,

[0382] - the first heavy chain comprises an anti-EGFR heavy chain variable region and a first heavy chain constant region;

[0383] - the first light chain comprises an anti-EGFR light chain variable region and a first light chain constant region;

[0384] - the second heavy chain comprises an anti-MET heavy chain variable region and a second heavy chain constant region;

[0385] The second light chain comprises an anti-MET light chain variable region and a second light chain constant region.

[0386] In some embodiments, the first and second heavy chain constant regions are heavy chain constant regions as defined above, preferably human IgG1 constant regions; the first and second light chain constant regions are light chain constant regions as defined above, preferably human kappa light chain constant regions. In some embodiments, the first heavy chain constant region comprises or consists of the amino acid sequence of SEQ ID NO: 65, and the second heavy chain constant region comprises or consists of the amino acid sequence of SEQ ID NO: 66. In some embodiments, the first and second light chain constant regions each comprise or consist of the amino acid sequence of SEQ ID NO: 12.

[0387] In some embodiments, the Ab unit of the ADC of the present disclosure is a full-length antibody comprising a heavy chain constant region and a light chain constant region. In some embodiments, the Ab unit has a tetrameric structure formed by two light chains and two heavy chains. In further embodiments, the Ab unit is an IgG antibody, particularly an IgG1 antibody.

[0388] In some preferred embodiments, the Ab unit of the ADC of the present disclosure comprises a first heavy chain, a first light chain, a second heavy chain, and a second light chain, wherein:

[0389] The first heavy chain comprises, or consists of, the amino acid sequence of SEQ ID NO:45, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto; and the first light chain comprises, or consists of, the amino acid sequence of SEQ ID NO:46, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto;

[0390] The second heavy chain comprises, or consists of, the amino acid sequence of SEQ ID NO:47, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto; and the second light chain comprises, or consists of, the amino acid sequence of SEQ ID NO:48, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0391] In some more preferred embodiments, the Ab unit of the ADC of the present disclosure comprises:

[0392] (a) an anti-EGFR heavy chain comprising the amino acid sequence of SEQ ID NO: 45 and an anti-EGFR light chain comprising the amino acid sequence of SEQ ID NO: 46, and

[0393] (b) an anti-MET heavy chain comprising the amino acid sequence of SEQ ID NO:47 and an anti-MET light chain comprising the amino acid sequence of SEQ ID NO:48.

[0394] Cancers that can be treated with the disclosed ADCs targeting EGFR and MET are EGFR and / or MET-positive cancers, such as solid tumors and hematological tumors, including but not limited to: non-small cell lung cancer, breast cancer, lung cancer, colorectal cancer, pancreatic cancer, ovarian cancer, gastric cancer, prostate cancer, head and neck tumors, liver cancer, bladder cancer, melanoma, esophageal cancer, kidney tumors, thyroid cancer, cervical cancer, lymphoma, uterine cancer, skin tumors, colon cancer, sarcoma, leukemia, glioblastoma, etc.

[0395] Ab unit targeting Nectin4

[0396] In a particularly preferred embodiment, the antibody conjugates provided herein include an antibody or antigen-binding fragment that specifically binds to human nectin-4 (anti-nectin-4 antibody), i.e., the antibody portion specifically targets the tumor-associated antigen nectin-4. Therefore, in some aspects, the present disclosure provides an antibody-drug conjugate (ADC) comprising an antibody or antigen-binding fragment thereof that specifically binds to nectin-4 as the Ab unit of the ADC.

[0397] Nectin-4 (Nectin cell adhesion molecule 4), also known as poliovirus receptor-related (PRR) protein, is a type I transmembrane cell adhesion molecule belonging to the Nectin family. Nectin-4 forms physical connections between adjacent cells and is essential for intercellular communication, migration, and other important cellular processes. Nectin-4 protein is specifically expressed in the embryo and placenta, and is only expressed at low levels in a few normal adult tissues (including the skin), while it is abnormally highly expressed in tumor tissues. Overexpression of nectin-4 in a variety of tumor cells is used as a marker for cancer recurrence and metastasis and is associated with poor prognosis in a variety of cancers. Cancers that overexpress nectin4 include, for example, breast cancer, cervical cancer, ovarian cancer, gastric cancer, esophageal cancer, head and neck cancer, lung cancer, non-small cell lung cancer, melanoma, bladder cancer, thyroid cancer, as well as hepatocellular carcinoma and urothelial carcinoma. Due to the target-specific expression of nectin-4, nectin-4 has become a potential biomarker and a promising therapeutic target. See, e.g., Jeffrey L Wong et al., Expert Opin Biol Ther. 2021 May 24; 21(7): 863–873. doi: 10.1080 / 14712598.2021.1929168, “Targeting nectin-4 by antibody-drug conjugates for the treatment of urothelial carcinoma.”

[0398] Antibodies targeting human nectin-4 that can be used in the ADCs of the present disclosure can be prepared using antibody preparation processes known in the art. For example, anti-nectin-4 antibodies can be obtained by immunizing animals with human nectin-4 (UniProtKB Accession No. Q96NY8) or a polypeptide comprising the amino acid sequence of the nectin-4 extracellular domain, harvesting antibodies from the immunized animals, and purifying and preferably humanizing the antibodies. Furthermore, fully human anti-human nectin-4 antibodies can be obtained using yeast display libraries expressing human immunoglobulin sequences or transgenic animals.

[0399] In some embodiments, the antibody or antibody fragment thereof (e.g., antigen-binding fragment) that specifically binds to human nectin-4 can be selected from Enfortumab, LY4052031, 9MW2821, BAT8007, SBT6290 or an antibody fragment thereof, or other anti-human nectin4 antibodies that recognize the same epitope or compete for binding to human nectin4.

[0400] Exemplary antibodies targeting human Nectin4 that can be used in the ADC of the present disclosure can be antibodies or antigen-binding fragments comprising all six CDR sequences of an antibody selected from the following group (preferably, comprising heavy chain variable region and light chain variable region sequences of an antibody selected from the following group): Enfortumab, LY4052031, 9MW2821, BAT8007, SBT6290. The present disclosure also considers the anti-human Nectin 4 antibodies disclosed in the following documents: WO2012047724, WO2022228406, WO2022228563, WO2024088390, WO2024012536, WO2024038075, WO2024017992, US12049500B2, US11179473B2, US11292837B2, US10675357B2, CN119013302A.

[0401] Enfortumab is a monospecific, bivalent IgG antibody that targets nectin 4. It directly binds to nectin 4-expressing cancer cells and triggers internalization of the antibody. Its sequence is available at NCATS Inxight Drugs and under FDA registration numbers UNII U1HUE4W970 and CAS No. 1448664-46-7.

[0402] Enfortumab light chain amino acid sequence (SEQ ID NO: 67)

[0403] >Enfortumab heavy chain amino acid sequence (SEQ ID NO: 68)

[0404] Enfortumab light chain LCDR1 sequence (SEQ ID NO: 69)

[0405] Enfortumab light chain LCDR2 sequence (SEQ ID NO: 70)

[0406] Enfortumab light chain LCDR3 sequence (SEQ ID NO: 71)

[0407] Enfortumab heavy chain HCDR1 sequence (SEQ ID NO: 72)

[0408] Enfortumab heavy chain HCDR2 sequence (SEQ ID NO: 73)

[0409] >Enfortumab heavy chain HCDR3 sequence (SEQ ID NO: 74)

[0410] Enfortumab light chain variable region VL (SEQ ID NO: 75)

[0411] Enfortumab heavy chain variable region VH (SEQ ID NO: 76)

[0412] In one embodiment, the antibody portion used in the ADC of the present disclosure comprises all six CDR sequences of Enfortumab. In another embodiment, the antibody portion used in the ADC of the present disclosure comprises the heavy chain variable region sequence and the light chain variable region sequence of Enfortumab. In yet another embodiment, the antibody portion used in the ADC of the present disclosure comprises the heavy chain sequence and the light chain sequence of Enfortumab.

[0413] In some embodiments, the Ab unit of the ADC of the present disclosure comprises three CDRs of the heavy chain variable region (VH) sequence of SEQ ID NO: 76 and three CDRs of the light chain variable region (VL) sequence of SEQ ID NO: 75, and preferably, wherein the CDRs are defined according to Kabat or IMGT or a combination thereof.

[0414] In some embodiments, the Ab unit of the disclosed ADC comprises three heavy chain complementarity determining regions (HCDRs) and three light chain complementarity determining regions (LCDRs), wherein:

[0415] According to the Kabat definition, HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 72, HCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 73, HCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 74, LCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 69, LCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 70, and LCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 71.

[0416] In one embodiment, the Ab unit of the ADC of the present disclosure comprises a heavy chain variable region, wherein the heavy chain variable region comprises: the amino acid sequence shown in SEQ ID NO: 76, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0417] In one embodiment, the Ab unit of the ADC of the present disclosure comprises a light chain variable region, wherein the light chain variable region comprises: the amino acid sequence shown in SEQ ID NO: 75, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0418] In some preferred embodiments, the Ab unit of the ADC of the present disclosure comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 76, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 75.

[0419] In some embodiments, the Ab unit of the ADC of the present disclosure preferably further comprises a heavy chain constant region and / or a light chain constant region of an antibody. Preferably, the heavy chain constant region is a heavy chain constant region derived from a human immunoglobulin. Preferably, the light chain constant region is a light chain constant region derived from a human immunoglobulin. In some aspects, the heavy chain constant region contained in the Ab unit can be any isotype or subtype, such as a heavy chain constant region of an IgG1, IgG2, IgG3 or IgG4 isotype, and preferably an IgG1, IgG2 or IgG4 heavy chain constant region, especially a human IgG1 heavy chain constant region. In further aspects, the light chain constant region contained in the Ab unit can be a kappa light chain constant region or a lambda light chain constant region, especially a human kappa light chain constant region.

[0420] In some embodiments, the Ab unit of the ADC of the present disclosure comprises a human IgG1 heavy chain constant region. Preferably, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 23, or an amino acid sequence comprising at least one, two, or three, but not more than 20, 10, or 5 amino acid changes relative to the amino acid sequence of SEQ ID NO: 23, or a sequence having at least 95-99% identity with the amino acid sequence of SEQ ID NO: 23.

[0421] In some embodiments, the Ab unit of the ADC of the present disclosure comprises a human kappa light chain constant region. Preferably, the light chain constant region comprises the amino acid sequence of SEQ ID NO: 12, or an amino acid sequence comprising at least one, two, or three, but not more than 20, 10, or 5 amino acid changes relative to the amino acid sequence of SEQ ID NO: 12, or a sequence having at least 95-99% identity with the amino acid sequence of SEQ ID NO: 12.

[0422] In some embodiments, the Ab unit of the ADC of the present disclosure is a full-length antibody comprising a heavy chain constant region and a light chain constant region. In some embodiments, the Ab unit has a tetrameric structure formed by two light chains and two heavy chains. In further embodiments, the Ab unit is an IgG antibody, particularly an IgG1 antibody.

[0423] In some preferred embodiments, the Ab unit of the ADC of the present disclosure comprises a heavy chain and a light chain, wherein: the heavy chain comprises the amino acid sequence of SEQ ID NO: 68, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto, or consists thereof. In other preferred embodiments, the Ab unit of the ADC of the present disclosure comprises a heavy chain and a light chain, wherein: the light chain comprises the amino acid sequence of SEQ ID NO: 67, or an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity thereto.

[0424] In some more preferred embodiments, the Ab unit of the ADC of the present disclosure comprises:

[0425] (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 68, and

[0426] (b) A light chain comprising the amino acid sequence of SEQ ID NO: 67.

[0427] Cancers that can be treated with the disclosed ADCs targeting nectin-4 are nectin-4-positive cancers, including but not limited to breast cancer, cervical cancer, ovarian cancer, gastric cancer, esophageal cancer, head and neck cancer, lung cancer, non-small cell lung cancer, melanoma, bladder cancer, thyroid cancer, as well as hepatocellular carcinoma and urothelial carcinoma.

[0428] Drug P unit

[0429] The drug P unit of the antibody-drug conjugate is also referred to herein as the payload of the ADC drug.

[0430] In some embodiments, the drug P unit that can be used in the ADC of the present disclosure is a KRas mutation inhibitor, including but not limited to G12C mutation, G12D mutation, G12V mutation, G12A mutation, G12R mutation, G12S mutation and G13D mutation inhibitor. In some embodiments, the drug P unit that can be used in the ADC of the present disclosure is those KRas mutation inhibitors described in the PCT application PCT / CN2023 / 122129 previously completed and filed by the applicant, more specifically the KRas mutation inhibitors defined herein below.

[0431] In other embodiments, the drug P unit that can be used in the ADC of the present disclosure is a pan-Ras inhibitor that can inhibit almost all Ras subtypes regardless of the mutation status, such as some of the drug P units implemented or exemplified below in the present disclosure, including but not limited to P8, P9, P12, P13, P16-23, P25-37, etc.

[0432] Embodiment 1: The drug P unit that can be used in the ADC of the present disclosure is specifically a compound of formula (I), a stereoisomer, a tautomer, a stable isotope variant, a pharmaceutically acceptable salt or a solvate thereof,

[0433] in:

[0434] M is selected from N or C-R1;

[0435] M' is selected from N or C-R1';

[0436] R1 and R1' are each independently selected from H, halogen, CN, -C optionally substituted by halogen 1-6 Alkyl and optionally halogen-substituted -OC 1-6 alkyl;

[0437] R a is selected from H, halogen, CN, -C optionally substituted by halogen 1-6 Alkyl, optionally substituted by halogen -C 2-6 Alkynyl and -OC optionally substituted by halogen or deuterium 1-6 alkyl;

[0438] R b and R c Together with the N they are connected to form wherein X is selected from CH2, N and O;

[0439] R2 and R2' are each independently selected from H, OH or -C optionally substituted by halogen 1-6 Alkyl; or R2 and R2' attached to non-adjacent ring carbon atoms together form an intracyclic bridge -(CH2) 1-2 - or -CH2=CH2-; or R2 and R2' attached to the same ring carbon atom together with the ring carbon atom to which they are attached form a 4-6 membered spirocycloalkyl or a 4-6 membered spiroheterocycloalkyl containing 1 or 2 heteroatoms selected from N and O;

[0440] Ar is selected from

[0441] R3 is selected from H, -NH2, -NHC 1-6 Alkyl, -N(C 1-6 Alkyl)2 and -OC 1-6 alkyl;

[0442] R4 is selected from -CN, halogen, -NO2, -C optionally substituted by halogen 2-6 Alkynyl and optionally halogen-substituted -C 1-6 alkyl;

[0443] R5 is selected from H, -CN, halogen, -NO2 and halogen substituted -C 1-6 alkyl;

[0444] R6 is selected from H, halogen, CN, -C 1-6 Alkyl and -C 2-6 Alkynyl, where -C 1-6 Alkyl and -C 2-6 each alkynyl group is independently optionally substituted with halogen;

[0445] R7, R7', R8 and R8' are each independently selected from H, halogen, CN, -NO2 and -C optionally substituted by halogen 1-6 alkyl;

[0446] V and W are each independently selected from H, halogen, -C 1-6 Alkyl, OH and NH2;

[0447] Z is selected from O, N and CH2;

[0448] R is

[0449] R9 and R 10 Each independently selected from H, deuterium, -C 1-6 Alkyl and -(CH2) n -C 3-6 Cycloalkyl, wherein -C 1-6 Alkyl and -C 3- 6 cycloalkyl groups are each independently optionally substituted with deuterium, halogen or -OC 1-6 Alkyl substituted, or R9 and R attached to the same carbon atom 10 Together with the carbon atom to which they are attached, they form C 3-4 Cycloalkyl;

[0450] R 11 Selected from H, -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl and -(CH2) n -C 3-6 Cycloalkyl, where C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl or C 3-6 The cycloalkyl groups are each independently optionally substituted with deuterium, halogen, CN or -OC 1-6Alkyl substitution;

[0451] R 12 Selected from H, halogen, -CN, -OH, -NH2, -NHC 1-6 Alkyl, -N(C 1-6 Alkyl)2, -OC 1-6 Alkyl, -OC 3-6 Cycloalkyl, -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, -(CH2) n -C 3-6 Cycloalkyl and =C(R d )2, where R d are each independently selected from H, halogen and -C 1-6 Alkyl, where each occurrence of C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl or C 3-6 The cycloalkyl groups are each independently optionally substituted with halogen, CN or -OC 1-6 Alkyl substitution;

[0452] R 13 Selected from H, -C 1-6 Alkyl and -(CH2) n -C 3-6 Cycloalkyl, wherein -C 1-6 Alkyl and -C 3-6 The cycloalkyl groups are each independently optionally substituted with halogen or -OC 1-6 Alkyl substitution;

[0453] k is an integer selected from 0 or 1

[0454] m is selected from an integer from 0 to 6; and

[0455] n is an integer selected from 0 to 2;

[0456] Therefore, formula (I) can be specifically expressed as:

[0457] Embodiment 1.1: A compound of Formula (I) according to Embodiment 1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein M is C-R1.

[0458] Embodiment 1.1.1: A compound of Formula (I) according to Embodiment 1.1, wherein R1 is H, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof.

[0459] Embodiment 1.1.2: A compound of Formula (I) according to Embodiment 1.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R1 is halogen, eg, F, Cl.

[0460] Embodiment 1.1.3: A compound of Formula (I) according to Embodiment 1.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R1 is CN.

[0461] Embodiment 1.1.4: A compound of Formula (I) according to Embodiment 1.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R1 is -C 1-6 Alkyl, preferably -C 1-3 Alkyl, more preferably -C 1-3 Alkyl, most preferably -C substituted with 1-3 F 1- 3 alkyl, such as -CF3; examples of R1 include but are not limited to -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)(CH3), -CH2CH2CH2CH3, -CH2CH(CH3)CH3, -C(CH3)3, -CH2Cl, -CH2F, -CHF2, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, -C2F5.

[0462] Embodiment 1.1.5: A compound of Formula (I) according to Embodiment 1.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R1 is -OC optionally substituted with halogen 1-6 Alkyl, preferably -OC optionally substituted by halogen 1-3 Alkyl groups, such as -OC 1-3 Alkyl, such as -OCH3, -OCH2CH3.

[0463] Embodiment 1.1.6: A compound of Formula (I) according to Embodiment 1.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R1 is selected from halogen and -C 1-6 Alkyl, preferably halogen and -C substituted by halogen 1-3 Alkyl groups, such as F, Cl and -CF3.

[0464] Embodiment 1.1.7: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1.1 to 1.1.6, wherein R a For H.

[0465] Embodiment 1.1.8: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1.1 to 1.1.6, wherein R a is CN or halogen, such as F, Cl.

[0466] Embodiment 1.1.9: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1.1 to 1.1.6, wherein R a is -C optionally substituted by halogen 1-6 Alkyl is as generally or specifically defined in Embodiment 1.1.4.

[0467] Embodiment 1.1.10: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1.1 to 1.1.6, wherein R a is -C optionally substituted by halogen 2-6 Alkynyl, preferably -C 2-4 Alkynyl, more preferably -C 2-4 Alkynyl groups, such as but not limited to

[0468] Embodiment 1.1.11: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1.1 to 1.1.6, wherein R a is -OC optionally substituted by halogen or deuterium 1-6 Alkyl, preferably -OC optionally substituted by halogen or deuterium 1-3 Alkyl, more preferably -OC optionally substituted by deuterium 1-3 Alkyl, such as -OCH3, -OCD3, -OCH2CH3, most preferably -OCH3.

[0469] Embodiment 1.1.12: A compound of Formula (I) according to any one of Embodiments 1.1 to 1.1.11, a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof, wherein M' is N.

[0470] Embodiment 1.1.13: A compound of Formula (I), stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof according to any one of Embodiments 1.1 to 1.1.11 wherein M' is C-R1'.

[0471] Embodiment 1.1.14: A compound of formula (I) according to Embodiment 1.1.13, wherein R1′ is selected from halogen, such as F, Cl, preferably F, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof.

[0472] Embodiment 1.1.15: A compound of Formula (I) according to Embodiment 1.1.13, a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof, wherein R1' is CN.

[0473] Embodiment 1.1.16: A compound of formula (I) according to Embodiment 1.1.13, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R1' is selected from -C 1-6 Alkyl and optionally halogen-substituted -OC 1-6 Alkyl is as generally or specifically defined in each of Embodiments 1.1.4 and 1.1.5.

[0474] Embodiment 1.2: A compound of Formula (I) according to Embodiment 1, wherein M is N, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof.

[0475] Embodiment 1.2.1: A compound of Formula (I) according to Embodiment 1.2, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R a For H.

[0476] Embodiment 1.2.2: A compound of Formula (I) according to Embodiment 1.2, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R a CN, or R a is a halogen, such as F, Cl.

[0477] Embodiment 1.2.3: A compound of Formula (I) according to Embodiment 1.2, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R a is -C optionally substituted by halogen 1-6 Alkyl is as generally or specifically defined in Embodiment 1.1.4.

[0478] Embodiment 1.2.4: A compound of Formula (I) according to Embodiment 1.2, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R a is -C optionally substituted by halogen 2-6 Alkynyl, preferably -C 2-4 Alkynyl, more preferably -C 2-4 Alkynyl groups, such as but not limited to

[0479] Embodiment 1.2.5: A compound of Formula (I) according to Embodiment 1.2, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R a is -OC optionally substituted by halogen or deuterium 1-6 Alkyl, preferably -OC optionally substituted by halogen or deuterium 1-3 Alkyl, more preferably -OC optionally substituted by deuterium 1-3 Alkyl, such as -OCH3, -OCD3, -OCH2CH3, most preferably -OCH3.

[0480] Embodiment 1.2.6: A compound of Formula (I), stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof according to any one of Embodiments 1.2 to 1.2.5, wherein M' is N.

[0481] Embodiment 1.2.7: A compound of Formula (I), stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof according to any one of Embodiments 1.2 to 1.2.5, wherein M' is C-R1'.

[0482] Embodiment 1.2.8: A compound of formula (I) according to Embodiment 1.2.7, wherein R1′ is selected from halogen, such as F, Cl, preferably F, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof.

[0483] Embodiment 1.2.9: A compound of Formula (I) according to Embodiment 1.2.7, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R1' is CN.

[0484] Embodiment 1.2.10: A compound of formula (I) according to Embodiment 1.2.7, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R1' is selected from -C 1-6 Alkyl and optionally halogen-substituted -OC 1-6 Alkyl is as generally or specifically defined in each of Embodiments 1.1.4 and 1.1.5.

[0485] Embodiment 1.3: The compound of formula (I) of embodiment 1, its stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate, wherein the structural fragment For example, Specifically,

[0486] M is C-R1, wherein R1 is selected from halogen (preferably F or Cl) or -C 1-6 Alkyl (preferably halogen-substituted -C 1-3 Alkyl, more preferably -CF3), R a is H, and M' is C-R1' and R1' is halogen, preferably F; or M is N, R a Selected from H, -C 2-6 Alkynyl (preferably -C 2-4 Alkynyl, more preferably -C≡CH) and -OC 1-6 Alkyl (preferably -OC optionally substituted by deuterium 1-3 alkyl, more preferably -OCH3, -OCD3, -OCH2CH3), and M' is C-R1' and R1' is halogen, preferably F; specific examples include but are not limited to:

[0487] Embodiment 2.1: A compound of formula (I), a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof according to any one of Embodiments 1 to 1.3, wherein Ar is

[0488] Embodiment 2.1.1: A compound of formula (I) according to Embodiment 2.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R3 is H; or R3 is selected from -NH2, -NHC 1-6 Alkyl and N(C 1- 6 alkyl) 2, preferably selected from -NH2, -NHC 1-3 Alkyl and N(C 1-3 alkyl)2, most preferably -NH2.

[0489] Embodiment 2.1.2: A compound of Formula (I) according to Embodiment 2.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R3 is -OC 1-6 Alkyl, preferably -OC 1-3 Alkyl, such as -OCH3, -OCH2CH3.

[0490] Embodiment 2.1.3: A compound of formula (I), stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof according to any one of Embodiments 2.1 to 2.1.2, wherein R4 is CN.

[0491] Embodiment 2.1.4: A compound of formula (I) according to any one of Embodiments 2.1 to 2.1.2, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R4 is a halogen selected from F, Cl, Br, I; or R4 is NO2.

[0492] Embodiment 2.1.5: A compound of formula (I) according to any one of Embodiments 2.1 to 2.1.2, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R4 is -C optionally substituted with halogen 2-6 Alkynyl, preferably -C 2-4 Alkynyl groups, such as but not limited to

[0493] Embodiment 2.1.6: A compound of formula (I) according to any one of Embodiments 2.1 to 2.1.2, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R4 is -C optionally substituted with halogen 1-6 Alkyl, preferably -C 1-3 Alkyl groups, such as but not limited to -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)(CH3), -CH2CH2CH2CH3, -CH2CH(CH3)CH3, -C(CH3)3, -CH2Cl, -CH2F, -CHF2, -CF3, -CCl3, -CH2CH2F, -CH2CHF2, -CH2CF3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, -C2F5.

[0494] Embodiment 2.1.7: A compound of Formula (I) according to any one of Embodiments 2.1 to 2.1.6, a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof, wherein R7 and R7' are each H; or R7 and R7' are each halogen, preferably F.

[0495] Embodiment 2.1.8: A compound of formula (I) according to any one of Embodiments 2.1 to 2.1.6, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein one of R7 and R7' is H and the other is selected from halogen, CN and NO2, wherein the halogen is preferably F; for example, R7 is H and R7' is halogen, preferably F, or R7' is H and R7 is halogen, preferably F.

[0496] Embodiment 2.1.9: A compound of formula (I) according to any one of Embodiments 2.1 to 2.1.6, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein one of R7 and R7' is H and the other is selected from -C 1-6 Alkyl, preferably -C 1-3 Alkyl groups, such as but not limited to -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)(CH3), -CH2CH2CH2CH3, -CH2CH(CH3)CH3, -C(CH3)3, -CH2Cl, -CH2F, -CHF2, -CF3, -CCl3, -CH2CH2F, -CH2CHF2, -CH2CF3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, -C2F5.

[0497] Embodiment 2.1.10: A compound of formula (I) according to any one of Embodiments 2.1 to 2.1.6, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein one of R7 and R7' is selected from halogen, -NO2, CN and -C optionally substituted by halogen 1-6 alkyl, the other selected from -C 1-6 Alkyl, wherein -C 1-6 Alkyl is preferably -C 1-3 Alkyl groups, as specifically exemplified in Embodiment 2.1.9.

[0498] Embodiment 2.1.11: A compound of Formula (I) according to any one of Embodiments 2.1 to 2.1.10, a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof, wherein V is H.

[0499] Embodiment 2.1.12: A compound of Formula (I), stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof according to any one of Embodiments 2.1 to 2.1.10, wherein V is -OH, or V is -NH2.

[0500] Embodiment 2.1.13: A compound of formula (I) according to any one of Embodiments 2.1 to 2.1.10, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein V is halogen; or V is -C 1-6 Alkyl, preferably -C 1-3 alkyl.

[0501] Embodiment 2.1.14: A compound of Formula (I) according to Embodiment 2.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein Ar is Wherein R3 is H or halogen, R4 is selected from halogen, -C 2-6 Alkynyl (preferably -C 2-4 Alkynyl) and -C 1-6 Alkyl (preferably -C 1-3 Alkyl), such as but not limited to

[0502] Embodiment 2.1.15: A compound of Formula (I) according to Embodiment 2.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein Ar is Wherein R3 is selected from -NH2, -NHC 1-6 Alkyl and N(C 1-6 Alkyl) 2, preferably selected from -NH2, -NHC 1-3 Alkyl and N(C 1-3 Alkyl) 2, most preferably -NH2; R4 is selected from -CN, halogen, -NO2 and -C substituted by halogen 2-6 Alkynyl; V is selected from H, -C 1-6 Alkyl (preferably -C 1-3 alkyl) and halogen, R7 and R7' are each H, or each halogen, or one of them is H and the other is halogen or halogen-substituted C 1-6 Alkyl (preferably halogen-substituted -C 1-3 alkyl), or one of which is halogen and the other is halogen-substituted C 1-6 Alkyl (preferably halogen-substituted -C 1-3 Alkyl), wherein the halogen is preferably F;

[0503] Preferably, Ar is V is H, R7 and R7' are each H, or one of them is H and the other is halogen, wherein halogen is preferably F;

[0504] For example, but not limited to:

[0505] Embodiment 2.1.16: A compound of Formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 2.1 to 2.1.15, wherein The asterisked positions may have axial chirality as appropriate (e.g., depending on the values ​​of R1 and R1'), including or

[0506] Embodiment 2.2: A compound of formula (I), a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof according to any one of Embodiments 1 to 1.3, wherein Ar is

[0507] Embodiment 2.2.1: The compound of formula (I) of embodiment 2.2, its stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate, wherein R5 is H; or R5 is halogen selected from F, Cl, Br, I; preferably R5 is halogen, most preferably F; or R5 is NO2; or R5 is halogen-substituted -C 1-3 alkyl.

[0508] Embodiment 2.2.2: A compound of formula (I) according to Embodiment 2.2 or 2.2.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R6 is H; or R6 is a halogen selected from F, Cl, Br, I.

[0509] Embodiment 2.2.3: A compound of Formula (I) according to Embodiment 2.2 or 2.2.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R6 is -C optionally substituted with halogen 1-6 Alkyl, preferably -C 1-3 Alkyl, more preferably -C 1-3 Alkyl groups, such as but not limited to -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)(CH3), -CH2CH2CH2CH3, -CH2CH(CH3)CH3, -C(CH3)3, -CH2Cl, -CH2F, -CHF2, -CF3, -CCl3, -CH2CH2F, -CH2CHF2, -CH2CF3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, -C2F5.

[0510] Embodiment 2.2.4: A compound of Formula (I) according to Embodiment 2.2 or 2.2.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R6 is -C 2-6 Alkynyl, preferably -C 2-4 Alkynyl, more preferably -C 2-4 Alkynyl groups, such as but not limited to Preferred

[0511] Embodiment 2.2.5: A compound of Formula (I) according to any one of Embodiments 2.2 to 2.2.4, a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof, wherein R8 and R8' are each H; or R8 and R8' are each halogen, preferably F.

[0512] Embodiment 2.2.6: A compound of formula (I) according to any one of Embodiments 2.2 to 2.2.4, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein one of R8 and R8' is H and the other is selected from halogen, CN and NO2, wherein the halogen is preferably F.

[0513] Embodiment 2.2.7: A compound of formula (I) according to any one of Embodiments 2.2 to 2.2.4, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein one of R8 and R8' is H and the other is selected from -C 1-6 Alkyl, preferably -C 1-3 Alkyl is as generally or specifically defined in Embodiment 2.1.9.

[0514] Embodiment 2.2.8: A compound of formula (I) according to any one of Embodiments 2.2 to 2.2.4, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein one of R8 and R8' is selected from H, halogen, -NO2, CN and -C optionally substituted by halogen 1-6 alkyl, the other selected from -C 1-6 Alkyl, wherein -C 1-6 Alkyl is preferably -C 1-3 Alkyl is as generally or specifically defined in Embodiment 2.1.9.

[0515] Embodiment 2.2.9: A compound of Formula (I), stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof according to any one of Embodiments 2.2 to 2.2.8, wherein W is -OH.

[0516] Embodiment 2.2.10: A compound of Formula (I), stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof according to any one of Embodiments 2.2 to 2.2.8, wherein W is -NH2.

[0517] Embodiment 2.2.11: A compound of Formula (I) according to Embodiment 2.2, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein Ar is Wherein R5 is H or halogen, R6 is selected from halogen, -C 2-6 Alkynyl and -C 1-6 R8 and R8 'are each H; or R8 and R8 'are each halogen, or one of R8 and R8 'is H, the other is selected from halogen, CN and NO2, or one of R8 and R8 'is H, the other is selected from -C optionally substituted by halogen 1-6 Alkyl, or one of R8 and R8' is selected from H, halogen, -NO2, CN and -C optionally substituted by halogen 1- 6 alkyl, the other selected from -C 1-6 alkyl;

[0518] Preferably, Ar is Wherein R5 is halogen, preferably F, and R6 is selected from -C 2-6 Alkynyl (preferably -C 2-4 Alkynyl), such as but not limited to

[0519] Embodiment 2.2.12: A compound of Formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 2.2 to 2.2.11, wherein The * may have axial chirality as appropriate, as exemplified in the definition section above.

[0520] Embodiment 3.1: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof according to any one of Embodiments 1 to 2.2.12, wherein R b and R c Together with the N they are connected to form In which k is 0 and X is selected from CH2, N and O, that is, R b and R c Together with the N they are connected to form or

[0521] k is 1 and X is selected from CH2, N and O, that is, R band R c Together with the N they are connected to form

[0522] Embodiment 3.1.1: A compound of Formula (I) according to Embodiment 3.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R b and R c Together with the N they are connected to form wherein R2 and R2' are each independently selected from H, OH and -C optionally substituted by halogen 1-6 Alkyl, preferably OH and optionally substituted by halogen -C 1-6 Alkyl, more preferably selected from OH and -C 1-3 or R2 and R2' attached to the same ring carbon atom together with the ring carbon atom to which they are attached form a 4-6 membered spirocycloalkyl or a 4-6 membered spiroheterocycloalkyl containing 1 or 2 heteroatoms selected from N and O.

[0523] Embodiment 3.1.1.1: A compound of formula (I) according to embodiment 3.1.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R2 and R2' are each independently attached to any ring carbon atom, preferably More preferred For example Further, one of R2 and R2' is -OH and the other is -C 1-6 Alkyl, preferably -C 1-3 Alkyl, more preferably methyl; or R2 and R2' together with the ring carbon atoms to which they are attached form a 4-6 membered spirocycloalkyl or a 4-6 membered spiroheterocycloalkyl containing 1 or 2 heteroatoms selected from N and O;

[0524] Further, where chemically feasible, the substituents R2 and R2' may each be in a stereoisomeric form, such as the R or S configuration;

[0525] Specific examples include, but are not limited to:

[0526] Embodiment 3.1.2: A compound of Formula (I) according to Embodiment 3.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R b and R c Together with the N they are connected to form wherein R2 and R2' attached to non-adjacent ring carbon atoms together form an intracyclic bridge -CH2-, -CH2CH2-, or -CH2=CH2-;

[0527] Specific examples include Preferred

[0528] Embodiment 4.1: A compound of formula (I) according to any one of Embodiments 1 to 3.1.2, a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof, wherein Z is O; or Z is N; or Z is CH2; preferably Z is O.

[0529] Embodiment 5.1: A compound of formula (I), a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof according to any one of Embodiments 1 to 4.1, wherein R is More preferably

[0530] Embodiment 5.1.1: A compound of formula (I) according to any one of Embodiments 1 to 5.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R9 and R 10 Both are H; or one or both are deuterium.

[0531] Embodiment 5.1.1.1: A compound of formula (I) according to any one of Embodiments 1 to 5.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R9 and R 10 Each independently selected from H and -C 1-6 Alkyl, preferably -C 1-3 Alkyl, optionally deuterated, halogen or -OC 1-6 Alkyl substituted, for example, one of them is H and the other is a defined alkyl, or both are defined alkyl; alkyl groups such as, but not limited to, -CH3, -CD3, -CH2CH3, -CH2CH2CH3, -CH(CH3)(CH3), -CH2-OCH3, -CH2-O-CH2CH3, -CH2CH2-O-CH3, -CH2CH2-O-CH2CH3, -CH2F, -CH2Cl, -CHF2, -CF3, -CCl3, -CH2CH2F, -CH2CHF2, -CH2CF3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3.

[0532] Embodiment 5.1.1.2: A compound of formula (I) according to any one of Embodiments 1 to 5.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R9 and R 10 are each independently selected from H and -(CH2) n -C 3-6Cycloalkyl (preferably -C 3-6 cycloalkyl), wherein C 3-6 Cycloalkyl is optionally substituted with halogen or C 1-6 Alkoxy substituted, for example, one of which is H and the other is a cycloalkyl as defined; for example but not limited to

[0533] Embodiment 5.1.1.3: A compound of formula (I) according to any one of Embodiments 1 to 5.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein R9 and R 10 Together with the carbon atom to which they are attached, they form C 3-4 Cycloalkyl, for example cyclopropyl, cyclobutyl.

[0534] Embodiment 5.1.2: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1 to 5.1.1.3, wherein R 11 For H.

[0535] Embodiment 5.1.2.1: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof, according to any one of Embodiments 1 to 5.1.1.3, wherein R 11 -C 1-6 Alkyl, preferably -C 1-3 Alkyl, optionally substituted by deuterium, halogen, CN or -C 1-6 Alkoxy is substituted, preferably optionally with deuterium, halogen or -C 1-3 Alkoxy substitutions; for example, but not limited to -CH3, -CD3, -CH2CH3, -CH2CD3, -CH2CH2CH3, -CH(CH3)(CH3), -CH2CH2CH2CH3, -CH2CH(CH3)CH3, -C(CH3)3, -CH2-OCH3, -CH2-O-CH2CH3, -CH2CH2-O-CH3, -CH2CH2-O-CH2CH3, -CH(CH3)CH2-OC H3, -CH2CH(CH3)-OCH3, -CH2F, -CH2Cl, -CHF2, -CF3, -CH2CH2F, -CH2CHF2, -CH(CH3)F, -CH(CH3)CH2F, -CH2CH(CH3)F, -CH2CF3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, -C2F5, -CH2CN, -CH2CH2CN; more preferably, R 11 -C 1-3Alkyl groups wherein hydrogen atoms are optionally replaced by one or more deuterium isotopes, e.g., -CH3, -CD3.

[0536] Embodiment 5.1.2.2: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1 to 5.1.1.3, wherein R 11 -C 2-6 Alkenyl or -C 2-6 Alkynyl, preferably -C 2-4 Alkenyl or -C 2-4 Alkynyl, optionally substituted by halogen, CN or -C 1-6 Alkoxy substituted; for example, but not limited to vinyl, propenyl, ethynyl, each optionally substituted with halogen or -C 1-6 Alkoxy substitution.

[0537] Embodiment 5.1.2.3: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1 to 5.1.1.3, wherein R 11 -(CH2) n -C 3-6 Cycloalkyl, preferably -C 3-6 Cycloalkyl, the -C 3-6 The cycloalkyl group is optionally substituted with deuterium, halogen, CN or -C 1-6 Alkoxy substitution; for example but not limited to

[0538] Embodiment 5.1.3: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1 to 5.1.2.3, wherein R 12 For H.

[0539] Embodiment 5.1.3.1: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof, according to any one of Embodiments 1 to 5.1.2.3, wherein R 12 is halogen, such as F, Cl, Br, I, preferably F; or R 12 For CN.

[0540] Embodiment 5.1.3.2: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1 to 5.1.2.3, wherein R 12 -NH2, -NHC 1-6 Alkyl, -N(C 1-6 alkyl)2, wherein -C1-6 The alkyl group is preferably -C 1-3 Alkyl, optionally substituted by halogen, CN or -OC 1-6 Alkyl substitution.

[0541] Embodiment 5.1.3.3: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1 to 5.1.2.3, wherein R 12 is -OH; or R 12 For-OC 1-6 Alkyl, preferably -OC 1-3 Alkyl, wherein the alkyl group is optionally substituted by halogen, CN or -OC 1-6 Alkyl substitution; for example, but not limited to -O-CH3, -O-CH2CH3, -O-CH2CH2CH3, -O-CH(CH3)(CH3), -O-CH2CH2CH2CH3, -O-CH2CH(CH3)CH3, -OC(CH3)3, -O-CH2Cl, -O-CH2CN, -O-CH2F, -O-CHF2, -O-CF3, -O-CCl3, -O-CH2CH2F , -O-CH2CH2CN, -O-CH2CHF2, -O-CH2CF3, -O-CH2CH2CH2F, -O-CH2CH2CHF2, -O-CH2CH2CF3, -OC( CH3)2CF3, -O-C2F5, -O-CH2-OCH3, -O-CH2-O-CH2CH3, -O-CH2CH2-O-CH3, -O-CH2CH2-O-CH2CH3.

[0542] Embodiment 5.1.3.4: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1 to 5.1.2.3, wherein R 12 -C 1-6 Alkyl, preferably -C 1-3 Alkyl, optionally substituted by halogen, CN or -OC 1-6Alkyl substitutions, such as but not limited to -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)(CH3), -CH2CH2CH2CH3, -CH2CH(CH3)CH3, -C(CH3)3, -CH2-OCH3, -CH2-O-CH2CH3, -CH2CH2-O-CH3, -CH2CH2-O-CH2CH3, -CH(CH3)CH2-OCH3, -CH2CH(CH3)-OCH3, -CH2F, -CH2Cl, -CHF2, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, -C2F5, -CH2CN, -CH2CH2CN.

[0543] Embodiment 5.1.3.5: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1 to 5.1.2.3, wherein R 12 For-OC 3-6 Cycloalkyl, wherein C 3- 6 Cycloalkyl is optionally substituted with halogen, CN or -OC 1-6 Alkyl substitution; for example but not limited to

[0544] Embodiment 5.1.3.6: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1 to 5.1.2.3, wherein R 12 -(CH2) n -C 3-6 Cycloalkyl, preferably -C 3-6 Cycloalkyl, the -C 3-6 Cycloalkyl is optionally substituted with halogen, CN or -C 1-6 Alkoxy substitution; as exemplified in Embodiment 5.1.2.3.

[0545] Embodiment 5.1.3.7: A compound of Formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof, according to any one of Embodiments 1 to 5.1.2.3, wherein R 12 -C 2-6 Alkenyl or -C 2-6 Alkynyl, preferably -C 2-4 Alkenyl or -C 2-4 Alkynyl, optionally substituted by halogen, CN or -C 1-6 Alkoxy substituted; for example, but not limited to vinyl, propenyl, ethynyl, each optionally substituted with halogen or -C1-6 Alkoxy substitution.

[0546] Embodiment 5.1.3.8: A compound of Formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1 to 5.1.2.3, wherein R 12 =C(R d )2, where R d Each independently selected from H, F, Cl, Br, I, -C 1-6 Alkyl (preferably -C 1-3 alkyl); for example, but not limited to, =CH2, =CF2, =CHF, =CCl2, =C(CH3)2, =C(CF3)2.

[0547] Embodiment 5.1.3.9: A compound of Formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1 to 5.1.2.3, wherein R 12 Selected from -C optionally substituted by halogen 1- 6 alkyl (preferably -C 1-3 alkyl) and =C(R d )2, where R d are each independently selected from H and halogen (preferably F).

[0548] Embodiment 5.1.3.10: A compound of Formula (I) according to any one of Embodiments 1 to 5.1.2.3, a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof, wherein m is 0, 1 or 2, preferably 1.

[0549] Embodiment 5.1.3.11: A compound of Formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof, according to any one of Embodiments 1 to 5.1.2.3, wherein R 12 The attached ring carbon atoms may be chiral, having either the R or S configuration, as appropriate.

[0550] Embodiment 5.1.3.12: A compound of Formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 5.1.3.8 to 5.1.3.9, wherein R 12 =C(R d )2 may have cis-trans isomerism, including E-type and Z-type, with E-type isomer being preferred.

[0551] Embodiment 5.1.4: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof, according to any one of Embodiments 1 to 5.1.3.12, wherein R 13 For H.

[0552] Embodiment 5.1.4.1: A compound of Formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof, according to any one of Embodiments 1 to 5.1.3.12, wherein R 13 is halogen, preferably F

[0553] Embodiment 5.1.4.2: A compound of Formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof, according to any one of Embodiments 1 to 5.1.3.12, wherein R 13 -C 1-6 Alkyl, preferably -C 1-3 Alkyl, optionally substituted with halogen or -OC 1-6 Alkyl substitutions, such as but not limited to -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)(CH3), -CH2CH2CH2CH3, -CH2CH(CH3)CH3, -C(CH3)3, -CH2-OCH3, -CH2-O-CH2CH3, -CH2CH2-O-CH3, -CH2CH2-O-CH2CH3, -CH(CH3)CH2-OCH3, -CH2CH(CH3)-OCH3, -CH2F, -CH2Cl, -CHF2, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, -C2F5.

[0554] Embodiment 5.1.4.3: A compound of Formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof, according to any one of Embodiments 1 to 5.1.3.12, wherein R 13 -(CH2) n -C 3-6 Cycloalkyl, where C 3-6 The cycloalkyl group is optionally substituted with halogen or -OC 1-6 Alkyl substitution; for example but not limited to

[0555] Embodiment 5.1.4.4.: A compound of Formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof, according to any one of Embodiments 1 to 5.1.3.12, wherein R13 -C 1-6 Alkyl, preferably -C 1-3 Alkyl, more preferably -CH3.

[0556] Embodiment 5.2: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof, according to any one of Embodiments 1 to 5.1.4.4, wherein R is More preferably Among them R9 and R 10 Both are H, or one or both are deuterium; R 11 -C 1-3 Alkyl, wherein the hydrogen atoms are optionally replaced by one or more isotopes of deuterium; R 12 Selected from -C optionally substituted by halogen 1-6 Alkyl (preferably -C 1-3 alkyl) and =C(R d )2, where R d are each independently selected from H and halogen (preferably F); R 13 -C 1-6 Alkyl (preferably -C 1-3 alkyl); m is 1 or 2;

[0557] Specifically, R is where R 11 、R 12 and m have the meanings defined above, R 11 Examples of include, but are not limited to, methyl, ethyl, isopropyl, -CD3, -CH2CD3; and / or R 12 Examples include, but are not limited to, fluoromethyl, difluoromethyl, methyl, fluoromethylene, difluoromethylene, methylene.

[0558] Embodiment 5.3: A compound of formula (I), a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof according to any one of Embodiments 1 to 5.1.4.4, wherein R is Preferably Among them R9 and R 10 Both are H, or one or both are deuterium; R 11 -C 1-3 Alkyl, wherein the hydrogen atoms are optionally replaced by one or more isotopes of deuterium; R 12 Selected from -C optionally substituted by halogen 1-6 Alkyl (preferably -C 1-3 alkyl) and =C(R d )2, where R d are each independently selected from H and halogen (preferably F); R 13 -C1-6 Alkyl (preferably -C 1-3 alkyl); m is 1 or 2;

[0559] Specifically, R is where R 11 、R 12 and m have the meanings defined above, R 11 Examples of include, but are not limited to, methyl, ethyl, isopropyl, -CD3, -CH2CD3; and / or R 12 Examples include, but are not limited to, fluoromethyl, difluoromethyl, methyl, fluoromethylene, difluoromethylene, methylene.

[0560] Embodiment 5.4: A compound of formula (I) according to any one of Embodiments 1 to 5.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein examples of R include but are not limited to

[0561] Preferred

[0562] Embodiment 6.1: The compound of formula (I) according to Embodiment 1, its stereoisomers, tautomers, stable isotopic variants, pharmaceutically acceptable salts or solvates, which has the sub-formula shown in Table 1 below:

[0563] wherein each substituent has the meaning generally or specifically defined above for each corresponding embodiment, and any combination of the generally or specifically defined meanings of each substituent is also encompassed;

[0564] Preferably, wherein

[0565] When M is C-R1, R1 is selected from halogen (preferably F or Cl) or -C 1-6 Alkyl (preferably halogen-substituted -C 1- 3 alkyl, more preferably -CF3), R a is H, and M' is C-R1' and R1' is halogen (preferably F);

[0566] When M is N, R a Selected from H, -C 2-6 Alkynyl (preferably -C 2-4 Alkynyl, more preferably -C≡CH) and -OC 1-6 Alkyl (preferably -OC optionally substituted by deuterium 1-3alkyl, more preferably -OCH3, -OCD3, -OCH2CH3), and M' is C-R1' and R1' is halogen (preferably F);

[0567] R2 and R2 attached to non-adjacent ring carbon atoms ’ Together they form an intra-ring bridge -(CH2) 1-2 - or -CH2=CH2-;

[0568] R3 is selected from -NH2, -NHC 1-6 Alkyl and N(C 1-6 Alkyl)2 (preferably selected from -NH2, -NHC 1-3 Alkyl and N(C 1-3 Alkyl) 2, more preferably -NH2); R4 is selected from -CN, halogen, -NO2 and -C substituted by halogen 2-6 Alkynyl; V is selected from H, -C 1-6 Alkyl (preferably -C 1-3 R7 and R7' are each H, or each halogen, or one of them is H and the other is halogen or halogen-substituted C 1-6 Alkyl (preferably halogen-substituted -C 1-3 alkyl), or one of which is halogen and the other is halogen-substituted C 1- 6 alkyl (preferably halogen-substituted -C 1-3 Alkyl), wherein the halogen is preferably F;

[0569] Preferably R3 is -NH2, R4 is -CN, V is H, R7 and R7' are each H, or one of them is H and the other is halogen (preferably F);

[0570] R5 is H or halogen (preferably F); R6 is selected from halogen, -C 2-6 Alkynyl (preferably -C 2-4 Alkynyl) and -C 1-6 Alkyl (preferably -C 1-3 R8 and R8' are each H, or R8 and R8' are each halogen, or one of R8 and R8' is H, the other is selected from halogen, CN and NO2, or one of R8 and R8' is H, the other is selected from -C optionally substituted by halogen 1-6 Alkyl, or one of R8 and R8' is selected from H, halogen, -NO2, CN and -C optionally substituted by halogen 1-6 alkyl, the other selected from -C 1-6 Alkyl; W is OH;

[0571] Preferably R5 is halogen (preferably F), R6 is selected from -C 2-6 Alkynyl (preferably -C 2-4alkynyl), R8 and R8' are each H, and W is OH;

[0572] Z is selected from O, N and CH2;

[0573] R9 and R 10 Both are H; or one or both are deuterium;

[0574] R 11 -C 1-6 Alkyl (preferably -C 1-3 alkyl) wherein the hydrogen atoms are optionally replaced by one or more isotopes of deuterium;

[0575] R 12 Selected from -C optionally substituted by halogen 1-6 Alkyl (preferably -C 1-3 alkyl) and =C(R d )2, where R d are each independently selected from H and halogen (preferably F);

[0576] m is 0, 1 or 2 (preferably 1 or 2);

[0577] R 13 -C 1-6 Alkyl (preferably -C 1-3 alkyl, more preferably -CH3).

[0578] Embodiment 6.2: The compound of formula (I) according to Embodiment 1, its stereoisomers, tautomers, stable isotopic variants, pharmaceutically acceptable salts or solvates, which has the sub-formula shown in Table 2 below:

[0579] wherein each substituent has the meaning generally or specifically defined above for each corresponding embodiment, and any combination of the generally or specifically defined meanings of each substituent is also encompassed;

[0580] Preferably, wherein

[0581] When M is C-R1, R1 is selected from halogen (preferably F or Cl) or -C 1-6 Alkyl (preferably halogen-substituted -C 1- 3 alkyl, more preferably -CF3), R a is H, and M' is C-R1' and R1' is halogen (preferably F);

[0582] When M is N, R a Selected from H, -C 2-6 Alkynyl (preferably -C 2-4 Alkynyl, more preferably -C≡CH) and -OC 1-6Alkyl (preferably -OC optionally substituted by deuterium 1-3 alkyl, more preferably -OCH3, -OCD3, -OCH2CH3), and M' is C-R1' and R1' is halogen (preferably F);

[0583] Structural fragments Selected from wherein X is selected from C, N and O; one of R2 and R2' is -OH and the other is -C 1-6 Alkyl, or R2 and R2' together with the ring carbon atoms to which they are attached form a 4-6 membered spirocycloalkyl or a 4-6 membered spiroheterocycloalkyl containing 1 or 2 heteroatoms selected from N and O;

[0584] Preferred Selected from wherein one of R2 and R2' is -OH and the other is -C 1- 3 alkyl, preferably methyl, more preferably

[0585] R3 is selected from -NH2, -NHC 1-6 Alkyl and N(C 1-6 Alkyl)2 (preferably selected from -NH2, -NHC 1-3 Alkyl and N(C 1-3 Alkyl) 2, more preferably -NH2); R4 is selected from -CN, halogen, -NO2 and -C substituted by halogen 2-6 Alkynyl; V is selected from H, -C 1-6 Alkyl (preferably -C 1-3 R7 and R7' are each H, or each halogen, or one of them is H and the other is halogen or halogen-substituted C 1-6 Alkyl (preferably halogen-substituted -C 1-3 alkyl), or one of which is halogen and the other is halogen-substituted C 1- 6 alkyl (preferably halogen-substituted -C 1-3 Alkyl), wherein the halogen is preferably F;

[0586] Preferably R3 is -NH2, R4 is -CN, V is H, R7 and R7' are each H, or one of them is H and the other is halogen (preferably F);

[0587] R5 is H or halogen (preferably F); R6 is selected from halogen, -C 2-6 Alkynyl (preferably -C 2-4 Alkynyl) and -C 1-6 Alkyl (preferably -C 1-3R8 and R8' are each H, or R8 and R8' are each halogen, or one of R8 and R8' is H, the other is selected from halogen, CN and NO2, or one of R8 and R8' is H, the other is selected from -C optionally substituted by halogen 1-6 Alkyl, or one of R8 and R8' is selected from H, halogen, -NO2, CN and -C optionally substituted by halogen 1-6 alkyl, the other selected from -C 1-6 Alkyl; W is OH;

[0588] Preferably R5 is halogen (preferably F), R6 is selected from -C 2-6 Alkynyl (preferably -C 2-4 alkynyl), R8 and R8' are each H, and W is OH;

[0589] Z is selected from O, N and CH2;

[0590] R9 and R 10 Both are H; or one or both are deuterium;

[0591] R 11 -C 1-6 Alkyl (preferably -C 1-3 alkyl) wherein the hydrogen atoms are optionally replaced by one or more isotopes of deuterium;

[0592] R 12 Selected from -C optionally substituted by halogen 1-6 Alkyl (preferably -C 1-3 alkyl) and =C(R d )2, where R d are each independently selected from H and halogen (preferably F);

[0593] m is 0, 1 or 2 (preferably 1 or 2);

[0594] R 13 -C 1-6 Alkyl (preferably -C 1-3 alkyl, more preferably -CH3).

[0595] Embodiment 6.3: A compound of formula (I'), a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, having the sub-formula shown in Table 2-1 below:

[0596] in

[0597] Among them, M, M', X, Z, V, W, R1, R1', R3, R4, R5, R6, R7, R7', R8, R8', R9, R 10 、R11 、R 12 、R 13 , k, m, n are as defined above for each embodiment of the compound of formula (I);

[0598] Structural fragments As defined above in any one of Embodiments 2.1 to 2.1.16 for compounds of formula (I);

[0599] Structural fragments As defined above in any one of Embodiments 2.2 to 2.2.12 for compounds of formula (I);

[0600] Structural fragments As defined above for any one of Embodiments 5.1 to 5.4 of the compounds of formula (I);

[0601] Y is selected from -CH2-, -CH2CH2-, -CH2OCH2- or is absent;

[0602] R 14 and R 15 Each independently selected from H and -C 1-6 alkyl.

[0603] Embodiment 6.3.1: A compound of Formula (I') according to Embodiment 6.3, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein:

[0604] exist In the fragment, when M is C-R1, R1 is selected from halogen (preferably F or Cl) or -C 1-6 Alkyl (preferably halogen-substituted -C 1-3 alkyl, more preferably -CF3), and M' is C-R1' and R1' is halogen (preferably F); or when M is N, M' is C-R1' and R1' is halogen (preferably F);

[0605] exist In the fragment, both M are C-R1, or one M is N and the other M is C-R1, and R1 is independently selected from H, halogen (preferably F) and halogen-substituted -C 1-6 Alkyl (preferably one or more F substituted -C 1-3 Alkyl, more preferably -CF3), preferably the M at the ortho position of R3 is selected from N and C-R1 (preferably C-halogen, more preferably CF), and the M at the ortho position of R6 is C-R1 (preferably C-halogen substituted -C 1-3 Alkyl, more preferably -C-CF3); R3 is selected from -NH2, -NHC 1-6 Alkyl and N(C 1-6Alkyl)2 (preferably selected from -NH2, -NHC 1-3 Alkyl and N(C 1-3 alkyl) 2, more preferably -NH2); R5 is H or halogen (preferably F); R6 is selected from halogen (preferably Cl) and -C 1-6 Alkyl (preferably -C 1-3 alkyl);

[0606] exist In the fragment, R3 is selected from -NH2, -NHC 1-6 Alkyl and N(C 1-6 Alkyl)2 (preferably selected from -NH2, -NHC 1-3 Alkyl and N(C 1-3 Alkyl) 2, more preferably -NH2); R4 is selected from -CN, halogen, -NO2 and -C substituted by halogen 2-6 Alkynyl; V is selected from H, -C 1-6 Alkyl (preferably -C 1-3 R7 and R7' are each H, or each halogen, or one of them is H and the other is halogen or halogen-substituted C 1-6 Alkyl (preferably halogen-substituted -C 1-3 alkyl), or one of which is halogen and the other is halogen-substituted C 1-6 Alkyl (preferably halogen-substituted -C 1-3 Alkyl), wherein the halogen is preferably F;

[0607] Preferably R3 is -NH2, R4 is -CN, V is H, R7 and R7' are each H, or one of them is H and the other is halogen (preferably F);

[0608] exist In the fragment, R5 is H or halogen (preferably F); R6 is selected from halogen, -C 2-6 Alkynyl (preferably -C 2-4 Alkynyl) and -C 1- 6 alkyl (preferably -C 1-3 R8 and R8' are each H, or R8 and R8' are each halogen, or one of R8 and R8' is H, the other is selected from halogen, CN and NO2, or one of R8 and R8' is H, the other is selected from -C optionally substituted by halogen 1-6 Alkyl, or one of R8 and R8' is selected from H, halogen, -NO2, CN and -C optionally substituted by halogen 1-6 alkyl, the other selected from -C 1-6 Alkyl; W is OH;

[0609] Preferably R5 is halogen (preferably F), R6 is selected from -C 2-6 Alkynyl (preferably -C2-4 alkynyl), R8 and R8' are each H, and W is OH;

[0610] X is selected from O and NH;

[0611] Y is selected from -CH2CH2- and absent;

[0612] Z is selected from O;

[0613] R9 and R 10 Both are H; or one or both are deuterium;

[0614] R 11 -C 1-6 Alkyl (preferably -C 1-3 alkyl) wherein the hydrogen atoms are optionally replaced by one or more isotopes of deuterium;

[0615] R 12 Selected from -C optionally substituted by halogen 1-6 Alkyl (preferably -C 1-3 alkyl) and =C(R d )2, where R d are each independently selected from H and halogen (preferably F);

[0616] m is 0, 1 or 2 (preferably 1 or 2);

[0617] n and k are 0 or 1 (preferably 0);

[0618] R 13 -C 1-6 Alkyl (preferably -C 1-3 alkyl, more preferably -CH3);

[0619] R 14 and R 15 H or -C 1-3 Alkyl, more preferably -CH3.

[0620] Embodiment 6.4: A compound of Embodiments 6.1 to 6.3.1, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein the R fragment Selected from

[0621] Embodiment 6.5: A compound according to any one of Embodiments 6.1 to 6.4, a stereoisomer, a tautomer, a stable isotopic variant, a pharmaceutically acceptable salt or a solvate thereof, wherein the Ar portion, i.e., the left-hand fragment, as appropriate, has axial chirality, as shown below:

[0622] For example

[0623] For example

[0624] For example

[0625] Embodiment 7: A KRas mutation inhibitor compound selected from the following compounds, stereoisomers, tautomers, stable isotopic variants, pharmaceutically acceptable salts or solvates thereof,

[0626] or a pharmaceutically acceptable salt or solvate thereof.

[0627] It should be noted that the KRas mutation inhibitor compounds suitable for the ADC disclosed herein as defined above are The NH ring when present in the fragment and / or the OH ring when present on the side chain naphthalene ring or benzothiophene ring are bonded to the linker unit L of the ADC.

[0628] Embodiment 8: A Ras mutation inhibitor compound selected from the following compounds, stereoisomers, tautomers, stable isotopic variants, pharmaceutically acceptable salts or solvates thereof,

[0629] Wherein, each Ras mutation inhibitor compound is connected to the linker unit L of ADC through -NH2, -NH-, N or -OH in its structure; or selected from the Ras inhibitor compounds disclosed in the following patents, the representative compounds are as follows: Ras protein degrader

[0630] Ras molecular glue

[0631] Tricyclic / tetracyclic Ras inhibitors

[0632] Small molecule Ras inhibitors

[0633] Other types of small molecule Ras inhibitors

[0634] It should be noted that the Ras mutation inhibitor compounds disclosed herein encompass the above-mentioned independent embodiments or specific embodiments, as well as embodiments consisting of any combination or sub-combination of the above-mentioned embodiments or specific embodiments, and embodiments consisting of any combination of any of the above-mentioned preferred or exemplary embodiments.

[0635] It should be noted that the present disclosure covers ADC compounds formed by any of the above-mentioned general, specific or preferred Ras mutation inhibitor compounds, the linker units defined herein, and the antibody drug.

[0636] Connector unit L

[0637] In the ADC described herein, a Ras inhibitor, such as a KRas mutation inhibitor, is connected to an antibody or antigen-binding fragment via a linker unit. The linker unit forms a covalent bond with a Ras inhibitor, such as a KRas mutation inhibitor, at one position thereof and forms a covalent bond with an antibody or antigen-binding fragment at another position thereof. The linker unit can be monovalent relative to the Ras inhibitor, such as a KRas mutation inhibitor, so that they covalently link a single Ras inhibitor, such as a KRas mutation inhibitor, to a single site on an antibody or its fragment, or can be multivalent relative to the Ras inhibitor, such as a KRas mutation inhibitor, so that they covalently link more than one Ras inhibitor, such as a KRas mutation inhibitor, to a single site on an antibody or its fragment. As used herein, the expression "linker unit" is intended to include unconjugated, partially conjugated (i.e., only coupled to a Ras inhibitor, such as a KRas mutation inhibitor or only coupled to an Ab) and fully conjugated forms of the linker unit (i.e., coupled to a Ras inhibitor, such as a KRas mutation inhibitor and an Ab).

[0638] The number of Ras inhibitors, e.g., KRas mutant inhibitors, attached to the antibody or antigen-binding fragment thereof of the ADC can vary (referred to as the "drug-to-antibody ratio" or "DAR") and will be limited by the number of available attachment sites on the antibody or antigen-binding fragment thereof and the number of Ras inhibitors, e.g., KRas mutant inhibitors, attached to a single linker. In an ADC comprising multiple Ras inhibitors, e.g., KRas mutant inhibitors, each Ras inhibitor, e.g., KRas mutant inhibitor may be the same or different. ADCs with a DAR of 10 or even higher are contemplated as long as the ADC does not exhibit unacceptable levels of aggregation under conditions of use and / or storage. In some embodiments, the ADCs described herein may have a DAR in the range of about 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, or 1 to 4. In some embodiments, the ADCs described herein may have a DAR in the range of about 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, or 2 to 4. In certain specific embodiments, the DAR of the ADC may be about 1, 2, 3, or 4. In other specific embodiments, the DAR of the ADC may be about 5, 6, 7, or 8. In some specific embodiments, the DAR of the ADC may be about 1.

[0639] The linker unit L suitable for the ADC of the present disclosure can be any linker that can achieve the coupling of the drug of the present disclosure to the antibody. Suitably, the addition of the linker should ensure the sufficient stability of the ADC of the present disclosure in the circulatory system, should not be prematurely cleaved in the circulation to trigger off-target toxicity, and can provide rapid and effective release of KRas mutation inhibitors in the target site (e.g., tumor cells or tumor environment). For example, the linker unit can be stable to the extracellular environment and serum chemistry, or can include an intentionally unstable linker unit and can release Ras inhibitors, such as KRas mutation inhibitors, in the extracellular environment or tumor microenvironment.

[0640] In some embodiments, the Linker unit comprises a bond designed to release a Ras inhibitor, such as a KRas mutant inhibitor, upon internalization of the ADC within a cell. In some specific embodiments, the Linker unit comprises a bond designed to be cleaved and / or digested or otherwise specifically or non-specifically degraded within a cell.

[0641] In some embodiments, the linker unit in the ADC of Formula (X) of the present disclosure is a non-degradable linker. Examples of non-degradable linker units include, but are not limited to, thioether linkers, N-succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), and maleimidocaproyl (MC). Generally, such linkers are more stable, and ADCs containing such linkers must be internalized by cells, where lysosomal proteases degrade the antibody portion of the ADC, releasing the active pharmaceutical ingredient.

[0642] In still other embodiments, the linker unit in the ADC of Formula (X) of the present disclosure is a degradable linker unit comprising one or more chemically or enzymatically degradable bonds. Drug release from ADCs comprising such linkers is triggered by the properties of the cleavage site in the linker. Thus, the cleavage site of such linkers can be designed based on the characteristics of the target therapeutic site (e.g., tumor cell lysosomes and / or the tumor environment).

[0643] In some embodiments, the degradable linker unit comprises a chemically labile group that exploits the differential properties between plasma and certain cytoplasmic compartments, such as the acidic environment of endosomes or lysosomes or the high concentration of sulfhydryl groups (e.g., glutathione) in the cytosol; in some cases, the plasma stability of a linker comprising a chemically labile group can be increased or decreased by altering the steric hindrance near the group using substituents.

[0644] In some embodiments, the chemically labile group of the degradable linker unit is an acid-labile group that can remain intact during the neutral pH circulation of the blood and hydrolyze under acidic conditions to release the Ras inhibitor, such as the KRas mutant inhibitor, for example, in the acidic tumor environment or when internalized into the endosomal (pH 5.0-6.5) and lysosomal (pH 4.5-5.0) cell compartments. This pH-dependent release mechanism can be optimized by chemical modification to fine-tune the release of the Ras inhibitor, such as the KRas mutant inhibitor, for a specific pH. Examples of such acid-labile groups include hydrazone, hydrazine, acetal, orthoester, or imine groups.

[0645] In some embodiments, the degradable linker unit comprises a reducible group, such as a disulfide group. When the ADC is internalized into cells, this group is reduced due to the presence of a more reducing environment (e.g., reduced glutathione) in the cytosol, thereby releasing the drug. Tumor cells can be hypoxic due to irregular blood flow, leading to increased reductase activity and elevated glutathione concentrations, thus facilitating the selective release of the drug in tumor cells by the disulfide-containing linker.

[0646] In some embodiments, the degradable linker unit is an enzymatically degradable linker unit and is more stable in plasma and the extracellular environment than chemically unstable linkers. Such linkers can be based on peptides or include peptide regions, or non-peptide linkers, such as peptide mimetics, or carbohydrates, esters, and amides. Such linkers can be cleaved by tumor-specific enzymes, such as tumor-specific proteases that have increased abundance in tumors and / or tumor environments, including but not limited to lysosomal proteases, such as cathepsins (e.g., cathepsin B), legumain, MMP-2 / 9, plasmin, esterases, amidases, glutathione, and the like.

[0647] Generally, the enzymatically degradable linker unit can be composed of a self-immolative linker, a cleavable linker, an optional property-modifying unit, an optional linker unit, and an antibody linker portion. The self-immolative linker connects the drug P to the cleavable linker, promoting the release of the active drug molecule from the rest of the ADC, such as p-aminobenzyl, p-hydroxybenzyl, p-aminobenzyloxyacyl, p-hydroxybenzyloxyacyl, etc.; the cleavable linker, under an enzyme-based release mechanism, will include peptides or peptide analogs that can be recognized by the enzyme, esters (such as carbamates, sulfates), amides, disulfide-containing moieties, carbohydrates, etc.; the addition of a property-modifying unit may be beneficial to improve the properties of the ADC, such as stability in the blood circulation, the efficacy of the ADC at the target site, and optimizing the hydrophilicity of the ADC. For example, when the drug is highly hydrophobic, the addition of a PEG unit can be considered (but not required) to optimize the hydrophilicity of the ADC, such as reducing precipitation and aggregation; the antibody linker connects the antigen-targeting antibody or antigen-binding fragment to the rest of the conjugate, and has a functional group that can form a bond with a functional group on the antibody.

[0648] In one aspect, the linker unit L in the ADC of formula (X) of the present disclosure has the structure of the following formula (II): -A(S)-B(D 1 )-DE(D 1 )-G- (II),

[0649] in

[0650] A is a self-immolative linker;

[0651] S is an optional solubilizing sugar unit;

[0652] B is a cleavable linker, which is absent or selected from a peptide residue of 2-8 amino acids, preferably a dipeptide, tripeptide or tetrapeptide; an amide bond-containing fragment; a carbamate bond-containing fragment; a thioether bond-containing fragment;

[0653] D and D 1each independently an optional property-modifying unit, which, when present, is selected from polyethylene glycol (PEG), a hydrophilic peptide, a cyclodextrin unit, a polyamine, a polyamide, a polysaccharide, a dendrimer, and a bifunctional hydrocarbon chain;

[0654] E is an optional linker unit;

[0655] G is the antibody linker connected to Ab.

[0656] Accordingly, the composition of the ADC of the present disclosure can be expressed as: [PA(S)-B(D 1 )-DE(D 1 )-G] q -Ab.

[0657] On the other hand, the linker unit L in the ADC of formula (X) of the present disclosure has the structure of the following formula (II'): -JB(D 1 )-DE(D 1 )-G- (II'),

[0658] in

[0659] J is a self-immolative linker;

[0660] B is a cleavable linker, which is absent or selected from a peptide residue of 2-8 amino acids, preferably a dipeptide, tripeptide or tetrapeptide; an amide bond-containing fragment; a carbamate bond-containing fragment; a thioether bond-containing fragment;

[0661] D and D 1 each independently an optional property-modifying unit, which, when present, is selected from polyethylene glycol (PEG), a hydrophilic peptide, a cyclodextrin unit, a polyamine, a polyamide, a polysaccharide, a dendrimer, and a bifunctional hydrocarbon chain;

[0662] E is an optional linker unit;

[0663] G is the antibody linker connected to Ab.

[0664] Accordingly, the composition of the ADC of the present disclosure can be expressed as: [PJB(D 1 )-DE(D 1 )-G] q -Ab.

[0665] The following provides general, specific, or preferred embodiments for each possible component of the linker unit L. It should be noted that the present disclosure encompasses any combination of the general, specific, or preferred embodiments of each component with the general, specific, or preferred embodiments of any other component or components to obtain the linker unit L; accordingly, the present disclosure encompasses any combination of the linker unit obtained with the general, specific, or preferred embodiments of the antibody and drug moiety defined in this disclosure to obtain the ADC compound.

[0666] G—Antibody Linker

[0667] The role of the antibody linker is to connect the antigen-targeting antibody or antigen-binding fragment to the rest of the conjugate. It has functional groups that can form bonds with functional groups on the antibody.

[0668] In some embodiments, the antibody linker has a nucleophilic group that can interact with a reactive electrophilic group on the antibody to form a covalent bond between the antibody and the linker unit. The electrophilic groups on the antibody include, but are not limited to, aldehyde and ketone carbonyl groups, and the nucleophilic groups on the antibody linker include, but are not limited to, hydrazide, hydroxylamine, amino, hydrazine, thiosemicarbazone, carboxylic acid hydrazine, and aryl hydrazide.

[0669] In other embodiments, the antibody linker has an electrophilic group that can interact with a reactive nucleophilic group on the antibody to form a covalent bond between the antibody and the linker unit. The nucleophilic groups on the antibody include, but are not limited to, sulfhydryl, hydroxyl, or amino functional groups, and the electrophilic groups on the antibody linker include, but are not limited to, maleimide, haloacetamide groups, activated disulfides, active esters such as NHS esters or HOBt esters, haloformates, acyl halides, alkyl halides, or benzyl halides such as haloacetamides. In some embodiments, the bond formed between the antibody linker and the antibody is a thioether, amide, ester, carbamate, carbonate, urea, disulfide, or ether.

[0670] The G in the linker unit L of formula (II) or formula (II') of the ADC of the present disclosure has the following structure:

[0671] -G1-G2-G3-, where:

[0672] G1 is a bonding atom from Ab, such as an S, N, or C atom;

[0673] G2 is selected from:

[0674] 5-10 membered heterocyclyl containing 1 or 2 heteroatoms selected from N, S and O, wherein the ring carbon atoms are optionally oxidized;

[0675] ·

[0676] ·

[0677] ·

[0678] ·

[0679] ·

[0680] The left * indicates the connection point with G1, and the right * indicates the connection point with G1. Indicates the connection point with G3;

[0681] G3 is selected from a bond, -C 1-10 Alkylene-C(=O)-, -C 3-10 Alkynylidene-C(=O)-, -C 3-10 Alkenylene-C(=O)-, -C 1-10 Heteroalkylene-C(=O)-, -C 3-8 Cycloalkylene-C(=O)-, -OC 1-10 Alkylene-C(=O)-, -C 6-10 Arylene-C(=O)-, -C 1-10 Alkylene-C 6-10 Arylene-C(=O)-, -C 6-10 Arylene-C 1-10 Alkylene-C(=O)-, -C 1-10 Alkylene-C 3-8 Cycloalkylene-C(=O)-, -C 3-8 Cycloalkylene-C 1-10 Alkylene-C(=O)-, -C 3-8 Heterocyclylene-C(=O)-, -C 1-10 Alkylene-C 3-8 Heterocyclylene-C(=O)-, -C 3-8 Heterocyclylene-C 1-10 Alkylene-C(=O)-, -C 1-10 Alkylene-C(=O)-NH-C 1-10 Alkylene-OC 1-10 Alkylene-C(=O)-, -C 3-10 Alkynylidene-C(=O)-NH-C 1-10 Alkylene-OC 1-10 Alkylene-C(=O)-, -C 3-10 Alkenylene-C(=O)-NH-C 1-10 Alkylene-OC 1-10 Alkylene-C(=O)-, -C 1-10 Heteroalkylene-C(=O)-NH-C 1-10 Alkylene-OC1-10 Alkylene-C(=O)-, -C 3-8 Cycloalkylene-C(=O)-NH-C 1-10 Alkylene-OC 1-10 Alkylene-C(=O)-, -OC 1-10 Alkylene-C(=O)-NH-C 1-10 Alkylene-OC 1-10 Alkylene-C(=O)-, -C6- 10 Arylene-C(=O)-NH-C 1-10 Alkylene-OC 1-10 Alkylene-C(=O)-, -C 1-10 Alkylene-C 6-10 Arylene-C(=O)-NH-C 1-10 Alkylene-OC 1-10 Alkylene-C(=O)-, -C 6-10 Arylene-C 1-10 Alkylene-C(=O)-NH-C 1-10 Alkylene-OC 1-10 Alkylene-C(=O)-, -C 1-10 Alkylene-C 3-8 Cycloalkylene-C(=O)-NH-C 1-10 Alkylene-OC 1-10 Alkylene-C(=O)-, -C 3-8 Cycloalkylene-C 1-10 Alkylene-C(=O)-NH-C 1-10 Alkylene-OC 1-10 Alkylene-C(=O)-, -C 3-8 Heterocyclylene-C(=O)-NH-C 1-10 Alkylene-OC 1-10 Alkylene-C(=O)-, -C 1-10 Alkylene-C 3-8 Heterocyclylene-C(=O)-NH-C 1-10 Alkylene-OC 1-10 Alkylene-C(=O)-, -C 3-8 Heterocyclylene-C 1-10 Alkylene-C(=O)-NH-C 1-10 Alkylene-OC 1-10 Alkylene-C(=O)-, -C 1-10 Alkylene-NH-, -C 1-10 Heteroalkylene-NH-, -C 3-8 Cycloalkylene-NH-, -OC 1-10 Alkylene-NH-, -C 6-10 Arylene-NH-, -C1-10 Alkylene-C 6-10 Arylene-NH-, -C 6-10 Arylene-C 1-10 Alkylene-NH-, -C 1-10 Alkylene-C 3-8 Cycloalkylene-NH-, -C 3-8 Cycloalkylene-C 1-10 Alkylene-NH-, -C 3- 8-heterocyclylene-NH-, -C 1-10 Alkylene-C 3-8 Heterocyclylene-NH- and -C 3-8 Heterocyclylene-C 1-10 Alkylene-NH-, wherein each group connected to -C(=O)- or -NH- in G3 is optionally substituted by Bu, and G3 is connected to the linker unit E (when present) or the property adjustment unit D (when present, and E is not present) or the cleavable linker B (when both E and D are not present) through its -C(=O)- or -NH-, and the other end group is connected to G2.

[0682] In some embodiments, each group in G3 that is attached to -C(=O)- or -NH- is optionally substituted with a Bu group: H, deuterium, halogen, NO2, CN, -OR h 、-OR h 、-N(R h )2. -COR h 、-CO2R h 、-C-(O)C(O)R h 、-C(O)CH2C(O)R h 、-S(O)R h 、-SO2R h 、C(O)N(R h )2、-SO2N(R h )2、-OC(O)R h 、-N(R h )SO2R h and -C optionally substituted by a prostaglandin group 1-6 Alkyl, where R h is H or -C optionally substituted by halogen 1-6 Alkyl, or two R h The groups and the nitrogen to which they are attached together form a 4-7 membered heterocyclic group. Preferably, the substituent of G3 is an aminoalkyl moiety, such as -(CH2) 1-6 NH2, -(CH2) 1-6 NHR h or -(CH2) 1-6 N(R h)2, or two R attached to the same N atom h The groups together with the nitrogen to which they are attached form an azetidinyl, pyrrolidinyl or piperidinyl group.

[0683] In other embodiments, each group in G3 that is attached to -C(=O)- or -NH- is optionally substituted with a Bu group of the following formula (A): in,

[0684] T0 is -C 1-6 Alkylene-;

[0685] T is selected from -C(R a )2-、-O-、-NR a - or does not exist;

[0686] Q is selected from -CO-, -O-, -NR a - or does not exist;

[0687] U is selected from glycosyl or its derivatives, and contains Hydrophilic peptides;

[0688] R a Selected from H or -C 1-6 Alkyl, preferably H or -CH3;

[0689] Subunit selected from natural amino acid residues and unnatural amino acid residues;

[0690] p is an integer from 0 to 4, such as 0-2, 1-2, 2-4;

[0691] t is an integer from 0 to 20, for example, 0-8, 0-6, 0-4, 0-2, 1-20, 1-10, 2-10, 2-8, 2-6, 2-4, 4-14, 6-10, 6-12, 8-10, 8-14.

[0692] The sugar group or its derivative in the substituent carried by G3 of the antibody linker of the ADC of the present disclosure is selected from a monosaccharide group, a disaccharide group, an oligosaccharide group or a polysaccharide group or a derivative thereof, preferably a monosaccharide or a disaccharide or a derivative thereof as generally, specifically or preferably defined for the solubilizing sugar unit S in the "Linker Unit L" section herein, more preferably a monosaccharide or a disaccharide group or a derivative thereof as exemplified in Tables 3 and 4.

[0693] In some embodiments, U is a monosaccharide or disaccharide as defined herein or a derivative thereof and is linked by a glycosidic bond (-O-sugar residue); in other embodiments, U is an amino sugar as defined herein or a derivative thereof and is linked by an amino group, preferably an amino sugar derivative of a monosaccharide or disaccharide; in other embodiments, U is a sugar acid or uronic acid as defined herein or a derivative thereof and is linked by a carbonyl group.

[0694] In the hydrophilic peptide carried by the ADC of the present disclosure, the amino acid residues as subunits can be the conventional 20 natural amino acids. Accordingly, the value combination of R, R', and R" in the repeating subunit -CO-CR'R"-NR- corresponds to the corresponding values ​​of alanine (Ala), arginine (Arg), aspartic acid (Asp), asparagine (Asn), histidine (His), glycine (Gly), glutamic acid (Glu), glutamine (Gln), phenylalanine (Phe), lysine (Lys), leucine (Leu), serine (Ser), tyrosine (Tyr), threonine (Thr), isoleucine (Ile), proline (Pro), tryptophan (Trp), valine (Val), cysteine ​​(Cys), and methionine (Met). Preferably, the amino acid residues are selected from polar natural amino acids, such as arginine, serine, threonine, tyrosine, cysteine, aspartic acid, asparagine, glutamic acid, glutamine, lysine, histidine, glycine, and tryptophan. The hydrophilic peptide can be a repeat of a single unit or a mixed repeat of different subunits.

[0695] In the hydrophilic peptide carried by the ADC of the present invention, the amino acid residues as subunits may also be amino acids other than the conventional 20 natural amino acids, such as ornithine (Orn), β-alanine (β-Ala), citrulline (Cit), sarcosine (Sar), and those amino acids in which R, R', and R" in the repeating subunit -CO-CR'R"-NR- are different from the corresponding groups or fragments in the 20 amino acids, such as R, R', and R" are selected from alkyl, aryl, acyl, amide, ketone, azido, hydroxyl, sulfhydryl, hydrazine, cyano, quaternary ammonium, halogen, hydrazide, alkenyl, alkynyl, ether, thiol, seleno, sulfonic acid, sulfate, borate, phosphono, phosphate, heterocycle, enone, imine, aldehyde, ester, thioacid , hydroxylamine, amino, etc. or any combination thereof, or a group comprising an alkyl group, an aryl group, an acyl group, an amide group, a ketone group, an azido group, a hydroxyl group, a sulfhydryl group, a hydrazine group, a cyano group, a quaternary ammonium group, a halogen group, a hydrazide group, an alkenyl group, an alkynyl group, an ether, a thiol group, a selenoyl group, a sulfonic acid group, a sulfate group, a borate group, a phosphono group, a phosphate group, a heterocycle, an enone group, an imine group, an aldehyde, an ester, a thioacid, a hydroxylamine, an amino group, etc.; preferably, those amino acids in which R, R' and / or R" comprise a hydrophilic group, for example, R, R', R" are each independently carboxyl, sulfonic acid group, sulfate group, phosphate group, amino, amide group, quaternary ammonium group, ether group, sulfhydryl group or hydroxyl group, or a group comprising carboxyl, sulfonic acid group, sulfate group, phosphate group, amino, amide group, quaternary ammonium group, ether group, sulfhydryl group and / or hydroxyl group, for example, an alkyl group or an aryl group, for example, C 1-6 alkyl.

[0696] In some embodiments, the amino acid unit of the hydrophilic peptide comprises a natural amino acid repeating unit, and the amino acid is preferably selected from arginine, serine, threonine, tyrosine, cysteine, aspartic acid, asparagine, glutamic acid, glutamine, lysine, histidine, glycine, and tryptophan; in other embodiments, the hydrophilic peptide comprises an amino acid repeating unit other than the 20 natural amino acids listed above, such as ornithine (Orn), citrulline (Cit), and sarcosine (Sar); in other embodiments, the hydrophilic peptide is a mixed hydrophilic peptide comprising the natural amino acids and the unconventional amino acids.

[0697] In some embodiments, in the subunit In the embodiment, preferably, R, R', and R" are each independently H, -C1-6 alkyl, carboxyl, sulfonic acid, sulfate, phosphoric acid, amino, amide, quaternary ammonium, ether, sulfhydryl or hydroxyl, or a group containing a hydrophilic group such as carboxyl, sulfonic acid, sulfate, phosphoric acid, amino, amide, quaternary ammonium, ether, sulfhydryl and / or hydroxyl, such as aryl or C 1-6 More preferably, R, R', R" are each independently H, -C 1-6Alkyl, carboxyl, sulfonic acid, phosphoric acid, amino, amide, quaternary ammonium, sulfhydryl or hydroxyl, or a group containing a hydrophilic group such as carboxyl, sulfonic acid, phosphoric acid, amino, amide, quaternary ammonium, sulfhydryl and / or hydroxyl, such as an aryl or C 1-6 alkyl.

[0698] In some embodiments, the G3 portion of the antibody linker of the ADC of the present disclosure is substituted with a Bu group of formula (A), wherein U of formula (A) is generally or specifically defined above and includes hydrophilic peptides.

[0699] In some embodiments, the hydrophilic peptide is linked via the carboxyl terminus, and in other embodiments, the hydrophilic peptide is linked via the amino terminus.

[0700] It should be noted that the present disclosure involves hydrophilic peptides in the definition of multiple structural fragments, and their definitions are all subject to the definition given herein for the fragment of formula (A).

[0701] For the ADC of the present disclosure, the hydrophilic peptide carried is, for example but not limited to, or through the amino-terminus wherein R is as defined above, including but not limited to the amino residues of various amino acids listed herein for hydrophilic peptides, R a Selected from H or -C 1-6 Alkyl, preferably H or -CH3; preferably polysarcosine, polyarginine, polyglycine containing 4-14 units, more preferably polysarcosine containing 6-12 units.

[0702] It should be noted that each example of hydrophilic peptides is given below. Hydrophilic peptides, especially A specific example of wherein R on N in the repeating unit a When it is a methyl group, it represents the polysarcosine preferably carried by the ADC of the present disclosure.

[0703] In some embodiments, U is selected from and said hydrophilic peptide linked via the amino terminus, e.g. In other embodiments, U is selected from and said hydrophilic peptide linked via the carboxyl terminus, e.g. where R a Selected from H or -C 1-6 Alkyl, preferably H or -CH3.

[0704] In some embodiments, T0 is -C 1-4 Alkylene-, preferably -C1-2 Alkylene-, more preferably methylene.

[0705] In some embodiments, In the fragment, T is -O-, e.g. Or T is -C(R a )2-, for example Or T is -NR a -,For example or T does not exist, e.g.

[0706] In some embodiments, In the fragment, T does not exist, p is 0-4, for example 0-2, and Q is -CO-, i.e. Or T is -C(R a )2-, p is 0-4, such as 0-2, and Q is -CO-, that is For example but not limited to Further, in such embodiments, U is preferably selected from and hydrophilic peptides such as or an amino sugar or a derivative thereof as defined herein and linked with an amino group, preferably an amino sugar derivative of a monosaccharide or a disaccharide, more preferably an amino sugar or a derivative thereof as shown in Table 3 or Table 4.

[0707] In some embodiments, In the fragment, T is absent, p is 0-4, such as 0-2, and Q is selected from -O- or -NR a -,Right now Or T is -C(R a )2-, p is 0-4, such as 0-2, and Q is selected from -O- and -NR a -,Right now For example but not limited to Further, in such embodiments, U is preferably selected from sugar acids or derivatives thereof as defined herein and is linked to a carbonyl group, preferably sugar acid derivatives of monosaccharides or disaccharides, more preferably sugar acid derivatives shown in Table 3 or Table 4; and hydrophilic peptides such as

[0708] In some embodiments, Q is absent, Excerpts from wherein p is 0-4, preferably 1-4, such as 1-2, for example Further, in this type of embodiment, U can be a saccharide group or a derivative thereof and is linked by a glycosidic bond, for example The sugar residues are linked via ethylene glycol; or U is an amino sugar or a derivative thereof and is linked via an amino group, for example

[0709] In some embodiments, t is an integer from 1 to 20, for example, 2-10, 2-8, 2-6, 2-4, 4-10, 4-14, 6-10, 6-12, 8-10, 8-14.

[0710] In some embodiments, G1 is an S atom; in other embodiments, G1 is an N atom; in other embodiments, G1 is a C atom. In the ADCs disclosed herein, preferably G1 is a sulfur atom of Ab.

[0711] In some embodiments, G2 is a 5-10 membered heterocyclic group, preferably a 5-6 membered heterocyclic group containing 1 or 2 heteroatoms selected from N, O and S, wherein the ring carbon atoms are optionally oxidized, such as but not limited to The * on the left indicates the connection point with G1, and the wavy line on the right indicates the connection point with G3.

[0712] In some embodiments, G2 is selected from It is the acid-amide part formed after hydrolysis of the succinimide part.

[0713] In a preferred embodiment, G2 is a maleimido group In another preferred embodiment, G2 is

[0714] In some embodiments, G2 is selected from Preferably selected from

[0715] In some embodiments, G3 is selected from -C 1-10 Alkylene-C(=O)-, -C 3-10 Alkynylidene-C(=O)-, -C 3-10 Alkenylene-C(=O)-, -C 1-10 Heteroalkylene-C(=O)-, -C 3-8 Cycloalkylene-C(=O)-, -OC 1-10 Alkylene-C(=O)-, -C6- 10 Arylene-C(=O)-, -C 1-10 Alkylene-C 6-10 Arylene-C(=O)-, -C 6-10 Arylene-C 1-10Alkylene-C(=O)-, -C 1-10 Alkylene-C 3-8 Cycloalkylene-C(=O)-, -C 3-8 Cycloalkylene-C 1-10 Alkylene-C(=O)-, -C 3-8 Heterocyclylene-C(=O)-, -C 1-10 Alkylene-C 3-8 Heterocyclylene-C(=O)-, -C 3-8 Heterocyclylene-C 1-10 Alkylene-C(=O)-, -C 1-10 Alkylene-NH-, -C 1-10 Heteroalkylene-NH-, -C 3-8 Cycloalkylene-NH-, -OC 1-10 Alkylene-NH-, -C 6-10 Arylene-NH-, -C 1-10 Alkylene-C 6-10 Arylene-NH-, -C 6-10 Arylene-C 1-10 Alkylene-NH-, -C 1-10 Alkylene-C 3-8 Cycloalkylene-NH-, -C 3-8 Cycloalkylene-C 1-10 Alkylene-NH-, -C 3-8 Heterocyclylene-NH-, -C 1-10 Alkylene-C 3-8 Heterocyclylene-NH- and -C 3-8 Heterocyclylene-C 1-10 Alkylene-NH-; preferably selected from -C 1-10 Alkylene-C(=O)-, -C 3-10 Alkynylidene-C(=O)-, -C 3-10 Alkenylene-C(=O)-, -C 1-10 Heteroalkylene-C(=O)-, -C 1-10 Alkylene-C 6-10 Arylene-C(=O)-, -C 6-10 Arylene-C 1-10 Alkylene-C(=O)-, -C 1-10 Alkylene-C 3-8 Cycloalkylene-C(=O)-, -C 3-8 Cycloalkylene-C 1-10 Alkylene-C(=O)-, -C 1-10 Alkylene-C 3-8 Heterocyclylene-C(=O)-, -C 3-8 Heterocyclylene-C 1-10 Alkylene-C(=O)-; more preferably -C1-10 Alkylene-C(=O)-; wherein each group in G3 connected to -C(=O)- or NH is optionally substituted by Bu as defined above for G3, and G3 is connected to the linker unit E (when present) or the property adjustment unit D (when present, and E is not present) or the cleavable linker B (when both E and D are not present) through its -C(=O)- or NH, and the other end group is connected to G2.

[0716] In some embodiments, G3 is -C 1-10 Alkylene-C(=O)-, preferably -C 1-5 Alkylene-C(=O)-, wherein the alkylene moiety is optionally substituted with the above-mentioned Bu moiety.

[0717] In some embodiments, G3 is -C 1-10 Alkylene-NH, preferably -C 1-5 Alkylene-NH-, wherein the alkylene moiety is optionally substituted with a Bu moiety as described above.

[0718] In an exemplary embodiment, -G2-G3- may be the following groups:

[0719] For example

[0720] For example

[0721] wherein G3 and Bu are each as generally or preferably defined above;

[0722] G3′ and G3″ are each independently selected from -C 1-10 Alkylene, -C 3-10 Alkynylidene, -C 3-10 Alkenylene, -C 1-10 Heteroalkylene, -C 3-8 Cycloalkylene, -C 6-10 Arylene, -C 1-10 Alkylene-C 6-10 Arylene, -C 6-10 Arylene-C 1-10 Alkylene, -C 1-10 Alkylene-C 3-8 Cycloalkylene, -C 3-8 Cycloalkylene-C 1-10 Alkylene, -C 3-8 Heterocyclylene, -C 1-10 Alkylene-C 3-8 Heterocyclylene, -C 3-8 Heterocyclylene-C1-10 alkylene;

[0723] Preferably, G3' and G3" are each independently selected from -C 1-10 Alkylene, -C 3-10 Alkynylidene, -C 3-10 Alkenylene, -C 1-10 Heteroalkylene, -C 1-10 Alkylene-C 6-10 Arylene, -C 6-10 Arylene-C 1-10 Alkylene, -C 1-10 Alkylene-C 3-8 Cycloalkylene, -C 3-8 Cycloalkylene-C 1-10 Alkylene, -C 1-10 Alkylene-C 3-8 Heterocyclylene, -C 3-8 Heterocyclylene-C 1-10 alkylene;

[0724] More preferably, G3' is -C 1-10 Alkylene-, preferably -C 1-5 Alkylene-; G3" is -C 1-5 Alkylene-, preferably -C 1-2 Alkylene-; wherein G3' and G3" are optionally substituted with Bu moieties as generally or preferably defined above;

[0725] The left * indicates the connection point with G1, and the right * indicates the connection point with G1. Indicates the point of attachment to E (when present) or property modulating unit D (when present and E is absent) or cleavable linker B (when both E and D are absent).

[0726] In some embodiments, -G2-G3- is Where G3' is -C 1-10 Alkylene-, preferably -C 1-5 Alkylene-, Bu is absent or is a structural fragment of formula (A).

[0727] In a specific embodiment, -G2-G3- can be For example Or -G2-G3- can be For example wherein T0, T, Q, U, p and the fragment of formula (A) are as generally or specifically defined above for formula (A).

[0728] In a further specific embodiment, -G2-G3- is For example Where T0 is -C 1-4 Alkylene-(e.g. -C 1-2 alkylene-, such as methylene), T is absent and p is 0-4 (e.g. 0-2) and Q is selected from -O- or -NR a -, or T0 is -C 1-4 Alkylene-(e.g. -C 1-2 Alkylene-, such as methylene), T is -C(R a )2-, p is 0-4 (e.g., 0-2) and Q is -NR a - or -O-, and U is preferably selected from sugar acids (preferably monosaccharide or disaccharide sugar acids) or their derivatives and is linked to a carbonyl group, Hydrophilic peptides such as or where T0 is -C 1-4 Alkylene-(e.g. -C 1-2 alkylene-, such as methylene), T is absent and p is 0-4 (such as 0-2) and Q is -CO-, or T0 is -C 1-4 Alkylene-(e.g. -C 1-2 Alkylene-, such as methylene), T is -C(R a )2-, p is 0-4 (e.g., 0-2) and Q is -CO-, and U is preferably selected from amino sugars (preferably amino monosaccharides or amino disaccharides) or derivatives thereof and is linked to an amino group, or hydrophilic peptides such as or

[0729] Where T0 is -C 1-4 Alkylene-(e.g. -C 1-2 Alkylene-, such as methylene), T is selected from -C(R a )2-、-O-、-NR a - or absent, Q absent, p is 0-4 (e.g. 0-2, 1-2), and U is preferably selected from a glycosyl (preferably a monosaccharide or disaccharide) or a derivative thereof and linked with a glycosidic bond, or an amino sugar (preferably an aminomonosaccharide or an aminodisaccharide) or a derivative thereof and linked with an amino group;

[0730] wherein t is an integer from 1 to 20, for example, 2-10, 2-8, 2-6, 2-4, 4-10, 4-14, 6-10, 6-12, 8-10, 8-14.

[0731] In some embodiments, -G1-G2-G3- is selected from the following groups:

[0732] For example Specifically

[0733] For example Specifically wherein G3, G3' and Bu are generally or preferably defined above; the left side * indicates the connection point with the rest of the antibody, the right side Indicates the point of attachment to the Linker unit E (when present) or the Property Modifying Unit D (when present and E is absent) or the Cleavable Linker B (when both E and D are absent).

[0734] In some embodiments, -G1-G2-G3- is Where G3' is -C 1-10 Alkylene-, preferably -C 1-5 Alkylene-, Bu is absent or is a structural fragment of formula (A).

[0735] In a specific embodiment, -G1-G2-G3- can be For example Or -G1-G2-G3- can be For example wherein T0, T, Q, U, p and the fragment of formula (A) are as generally or specifically defined above for formula (A).

[0736] In a further specific embodiment, -G1-G2-G3- is For example Where T0 is -C 1-4 Alkylene-(e.g. -C 1-2 alkylene-, such as methylene), T is absent and p is 0-4 (e.g. 0-2) and Q is selected from -O- or -NR a -, or T0 is -C 1-4 Alkylene-(e.g. -C 1-2 Alkylene-, such as methylene), T is -C(R a )2-, p is 0-4 (e.g., 0-2) and Q is -NR a - or -O-, and U is preferably selected from sugar acids (preferably monosaccharide or disaccharide sugar acids) or their derivatives and is linked to a carbonyl group, Hydrophilic peptides such as or where T0 is -C 1-4 Alkylene-(e.g. -C 1-2 alkylene-, such as methylene), T is absent and p is 1-4 (such as 1-2) and Q is -CO-, or T0 is -C 1-4 Alkylene-(e.g. -C 1-2 Alkylene-, such as methylene), T is -C(Ra )2-, p is 0-4 (e.g., 0-2) and Q is -CO-, and U is preferably selected from amino sugars (preferably amino monosaccharides or amino disaccharides) or derivatives thereof and is linked to an amino group, or hydrophilic peptides such as or

[0737] Where T0 is -C 1-4 Alkylene-(e.g. -C 1-2 Alkylene-, such as methylene), T is selected from -C(R a )2-、-O-、-NR a - or absent, Q absent, p is 0-4 (e.g. 0-2, 1-2), and U is preferably selected from a glycosyl (preferably a monosaccharide or disaccharide) or a derivative thereof and linked with a glycosidic bond, or an amino sugar (preferably an aminomonosaccharide or an aminodisaccharide) or a derivative thereof and linked with an amino group;

[0738] wherein t is an integer from 1 to 20, for example, 2-10, 2-8, 2-6, 2-4, 4-10, 4-14, 6-10, 6-12, 8-10, 8-14.

[0739] In an exemplary embodiment, G (-G1-G2-G3-) in Formula (II) or Formula (II') of the present disclosure has the following structure:

[0740] In some preferred embodiments, G in Formula (II) or Formula (II') of the present disclosure has the following structure:

[0741] and wherein the maleimide moiety is The various fragments shown above, such as wait.

[0742] E—Connector unit

[0743] The linker unit E in the linker unit L of formula (II) or formula (II') of the ADC of the present disclosure connects the antibody linker G with the property-modifying unit D, or connects the antibody linker G with the cleavable linker B when D is absent, and is used to increase additional distance between the antibody linker and the cleavable linker B to potentially facilitate activation of B.

[0744] In some embodiments, the linker unit E is selected from a directly connected bond, a -C 1-5 Alkylene-, -NH-, -NH-C 1-5 Alkylene-heteroaryl (e.g., 5-membered or 6-membered nitrogen-containing heteroaryl, such as triazolyl), and the following groups:

[0745] wherein E, when present, is connected to the property-modifying unit D (when present) or the cleavable linker B via its -C(=O)- or -NH-terminus, as appropriate, and the other terminus is also connected to G3 via -C(=O)- or -NH-terminus, as appropriate;

[0746] R g Each independently selected from H and -C 1-6 Alkyl, preferably H or -C 1-3 alkyl;

[0747] R f Selected from -C 1-6 Alkylene-, -arylene-, -C 1-10 Heteroalkylene-, -C 3-8 Heterocyclylene-, -C 1-10 Alkylene-C 6-10 Arylene-, -C 6-10 Arylene-C 1-10 Alkylene-, -C 1-10 Alkylene-C 3-8 Cycloalkylene-, -C 3-8 Cycloalkylene-C 1-10 Alkylene-, -C 1-10 Alkylene-C 3-8 Heterocyclic-, -C 3-8 Heterocyclylene-C 1-10 Alkylene-;

[0748] It should be noted that, when present, each specific definition of E above can be repeated 1-10 times, preferably 1-4 times, such as 1 time, 2 times, 3 times and 4 times, end to end in different directions.

[0749] In a specific embodiment, the linker unit E is where R f Each independently selected from -C 1-6 Alkylene-, preferably -C 2-4 Alkylene, R g H or C 1-3 Alkyl groups, e.g. The wavy line on the left is connected to the cleavable linker B, and the wavy line on the right is connected to the antibody linker G.

[0750] In another specific embodiment, the linker unit E is a directly connected bond; in another specific embodiment, the linker unit E is -C 1-4 Alkylene-; In another specific embodiment, the linker unit E is -NH-.

[0751] D and D 1 —Property adjustment unit

[0752] The property-modifying unit D or D in the linker unit L of formula (II) or formula (II') of the ADC of the present disclosure 1 The addition of may be beneficial to improve the properties of ADC, such as stability in blood circulation, improved hydrophilicity, resulting in reduced clearance and increased exposure. However, D or D 1 An increase in the number of D2 will also lead to an increase in the molecular weight of the ADC and an increase in the hydrodynamic radius, which will lead to a decrease in diffusion. The decreased diffusion rate may reduce the ability of the ADC to penetrate into the tumor. Due to these two competing pharmacokinetic effects, a moderate D2 or D3 should be used. 1 The goal is to reduce the clearance of the ADC, thereby increasing plasma exposure, but not so much as to reduce its diffusivity as to interfere with the ADC's ability to reach the intended target cell population.

[0753] The property regulation unit D exists in the linker unit in a serial form. 1 It exists in a branched form. D can be directly connected between B and E, or can be connected between B and E through another connecting group, such as amino, carbonyl, alkylcarbonyl, amide, ester, urea, disulfide bridge, carbamate, hydrazone, imine, oxime, triazolyl, maleimide, alkenyl, alkynyl or alkylene. For example, D can be connected through -NH-, -C(=O)-, -C 1-4 Alkylene, -C 1-4 Alkylene-C(=O)-, -NH-C(=O)-(CH2OCH2)-C(=O)-, -C 1-4 Alkylene-NH-C(=O)-(CH2OCH2)-C(=O)- is attached to the cleavable linker B.

[0754] In some embodiments, D or D 1 Does not exist.

[0755] In some embodiments, D or D 1 Selected from polyamines, such as but not limited to polyethyleneimine, polylysine, spermine, dimeric polyamines, arginine, amidine, protamine, cationic lipids, cationic porphyrins, quaternary ammonium salts of polyamines, α-helical peptides.

[0756] In some embodiments, D or D 1 Selected from peptides, preferably hydrophilic peptides.

[0757] In some embodiments, D or D 1 It is a cyclodextrin unit.

[0758] In some embodiments, D or D 1 For polyamide.

[0759] In some embodiments, D or D 1It is a polysaccharide, a dendrimer or a bifunctional hydrocarbon chain.

[0760] In some embodiments, D or D 1 For PEG.

[0761] In the embodiments of the present disclosure, polydisperse PEG (a heterogeneous mixture of size and molecular weight), monodisperse PEG (a single chain length and molecular weight), and discrete PEG can be used as part of the property-modulating unit in the ADC of the present disclosure. Preferred PEGs are discrete PEGs, which are compounds synthesized in a stepwise manner rather than by a polymerization process, and are single molecules with a defined chain length.

[0762] The ADC of the present disclosure may comprise one or more PEG chains consisting of at least two ethylene oxide (CH2CH2O) subunits. The PEG chains may be linked together, for example, in a linear, branched, or star-shaped configuration. Typically, at least one PEG chain is functionalized so that it can be covalently linked to other components in the linker unit, and the functionalization method includes, for example, via an amine, thiol, NHS ester, maleimide, alkyne, azide, carbonyl, or other functional group. For example, one end of the PEG chain may be functionally derivatized to covalently link to an appropriate site of the linker unit, and the other end (or ends) may be free and unconstrained, i.e., not connected to other components in the ADC, and may take the form of a methoxyl group, a carboxylic acid, an alcohol, or other suitable functional group; or the PEG chain may be covalently linked in series between two components of two linker units, for example, between a linker unit E and a cleavable linker B. The connection between the PEG chain and the components within the linker unit is a cleavable bond, which may be a bond that is substantially insensitive to cleavage when circulating in plasma but sensitive to cleavage in the intracellular or intratumoral environment. Exemplary linkages include, but are not limited to, amide bonds, ether bonds, ester bonds, hydrazone bonds, oxime bonds, disulfide bonds, peptide bonds, or triazole bonds, among others.

[0763] Methods for attaching PEG units to the linker units of ADCs are well known to those skilled in the art. For example, PEG can be covalently bound to amino acid residues via reactive groups. Reactive groups are those to which activated PEG molecules can bind, such as free amino groups or carboxyl groups. For example, thiol groups on cysteine ​​residues can also serve as reactive groups for attaching PEG. In some embodiments, methoxylated PEG ("mPEG") with different reactive moieties can also be used to attach PEG to amino groups. Non-limiting examples of such reactive moieties include succinimidyl succinate (SS), succinimidyl carbonate (SC), mPEG-imidate, p-nitrophenyl carbonate (NPC), succinimidyl propionate (SPA), and cyanuric chloride. Accordingly, non-limiting examples of the mPEG include mPEG-succinimidylsuccinate (mPEG-SS), mPEG-succinimidyl carbonate (mPEG-SC), mPEG-imidate, mPEG-p-nitrophenyl carbonate (mPEG-NPC), mPEG-succinimidyl propionate (mPEG-SPA), mPEG-N-hydroxy-succinimide (mPEG-NHS), mPEG-cyanuric chloride, mPEG2-lysinol-NPC and mPEG2-Lys-NHS.

[0764] In some embodiments, the PEG unit comprises one or more linear PEG chains, each PEG chain having 2-12 ethylene oxide (CH2C2O) subunits, e.g., at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, or at least 12 ethylene oxide (CH2C2O) subunits. In preferred embodiments, the PEG unit comprises at least 2, at least 4, at least 6, at least 8, at least 10, or at least 12 ethylene oxide (CH2C2O) subunits, e.g., the PEG unit comprises 2 to 12, 2 to 10, 2 to 8, 2 to 6, 2 to 4, 4 to 12, 4 to 10, 4 to 8, 4 to 6, 6 to 12, 6 to 10, 6 to 8, 8 to 12, or 8 to 10 subunits.

[0765] Exemplary embodiments of the PEG fragment D linked to the cleavable linker B and the linker unit E include, but are not limited to, the following:

[0766] Exemplary embodiments of linear PEG units (i.e., one end connected to a linker unit and the other end being a terminal cap) are:

[0767] Wherein the left wavy line indicates the attachment site to E or G3 (or to E or B, as appropriate, in the case of branched PEG), the right wavy line indicates the attachment site to B, and each subscript c is independently selected from an integer between 2 and 12. In some embodiments, c is 2, 4, 8, or 12. In some embodiments, c is 2. In some embodiments, c is 4. In some embodiments, c is 8. In some embodiments, c is 12.

[0768] B—cleavable linker

[0769] In one aspect, the cleavable linker B in the linker unit L of Formula (II) or Formula (II') of the disclosed ADC is an enzymatically cleavable unit, i.e., a substrate that can be cleaved or broken by an enzyme. To limit or minimize the non-target toxicity of the ADC while ensuring the release of the toxin molecule at the target tumor site, the cleavable linker is designed to be a substrate specifically cleaved by an enzyme present near or within the target cell. Preferably, the level or activity of the enzyme near or within the target cell is higher than that in other parts of the body. This ensures that the ADC is specifically recognized, cleaved, and released by the enzyme in the target cell / tissue, thereby exerting its biological function.

[0770] Therefore, the ADCs of the present disclosure are stable in biological fluids until they reach their target, such as tumor cells / tissues. Due to the specific release of the ADCs, administration of the ADCs of the present disclosure will produce less toxicity than administration of a single KRas mutant inhibitor, and the activity of the KRas mutant inhibitor is maximized.

[0771] In some embodiments, B in the linker unit L of formula (II) or formula (II') of the ADC of the present disclosure is a dipeptide, tripeptide or oligopeptide consisting of an amino acid sequence specifically recognized and cleaved by a protease, preferably a non-mammalian protease or an endogenous mammalian protease present near or inside the target cell, such as but not limited to: β-site APP-cleaving enzyme 1 (BACE1), cathepsin D (CTSD), calpain-1 (CAPN1), aspartic endo-aspartate enzyme (Lugemain), caspase 1 (Casp1), caspase 2 (Casp2), caspase 3 ( Caspase 3), caspase 5 (CASPS), caspase 6 (Casp6), caspase 7 (Casp7), caspase 8 (Casp8), caspase 9 (Casp9), cathepsin B (CTSB), cathepsin K (CTSK), cathepsin L (CTSL), cathepsin S (CTSS), esterases (e.g., cholinesterase, alkaline phosphatase, phosphodiesterase, sulfatase), amidase angiotensin I converting enzyme (ACE), angiotensin I converting enzyme 2 (ACE2), ADAM metallopeptidase domain 10 (ADAM10), dipeptidyl peptidase 3 (DP3), phosphodiesterase (PS3), phosphodiesterase (PS4), phosphodiesterase (PS5), phosphodiesterase (PS6), phosphodiesterase (PS7), phosphodiesterase (PS8), phosphodiesterase (PS9), phosphodiesterase (PS1), phosphodiesterase (PS1), phosphodiesterase (PS2), phosphodiesterase (PS3 ... P3), insulin degrading enzyme (IDE), matrix metalloproteinase 1 (MMP1), matrix metalloproteinase 12 (MMP12), matrix metalloproteinase 13 (MMP13), matrix metalloproteinase 14 (membrane inserted) (MMP14), matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 3 (MMP3), matrix metalloproteinase 7 (MMP7), matrix metalloproteinase 8 (MMP8), matrix metalloproteinase 9 (MMP9), membrane metalloendopeptidase (NEPRILYSIN) (MME), ADAM metallopeptidase domain 17 (TACE) (ADAM17), dipeptidyl peptidase 4 (D PP4), dipeptidyl peptidase 8 (DPP8), dipeptidyl peptidase 9 (DPP9), coagulation factor Xa (factor Xa), coagulation factor VILA (factor VII), fibroblast activation protein α (FAP), furin (paired basic amino acid cleaving enzyme), granzyme A (granzyme 1, cytotoxic T lymphocyte-associated (GZMA), serine esterase 3), granzyme B (granzyme 2, cytotoxic T lymphocyte-associated (GZMB), serine esterase 1), granzyme K (granzyme 3, trypsin II) (GZMK), kallikrein-1 (KLK1), kallikrein-2 (KLK2), plasma kallikrein (PSA,KLK3), kallikrein-11 (KLK11), kallikrein-13 (KLK13), kallikrein-15 (KLK15), matrix enzyme (ST14), spinesin (TMPRSSS), plasmin (PLG), prolyl oligopeptidase (PREP), thrombin (F2), tPA, plasminogen activator tissue type (PLAT), UPA, plasminogen activator urokinase (PLAU), HtrA serine peptidase 2 (hTRA2), caseinolytic mitochondrial matrix peptidase proteolytic subunit (CIPP / X), constitutive proteasome chymotrypsin-like form (PSMBS, β5), constitutive proteasome trypsin-like form (PSMB7, β7), constitutive proteasome caspase-like form (PSMB6, β6), immunoproteasome chymotrypsin-like form (PSMB8, LMP7), immunoproteasome trypsin-like form (PSMB, 10 , MECL1), immunoproteasome caspase-like forms (PSMB9, LMP2).

[0772] In some embodiments, B in the linker unit L of formula (II) or formula (II') of the ADC of the present disclosure is specifically recognized and cleaved by cathepsin B, such as Cit-Val, Ala-Val, Gly-Phe-Gly-Gly, or Gly-Phe-Leu-Gly.

[0773] In some embodiments, B in the linker unit L of formula (II) or formula (II') of the ADC of the present disclosure is specifically recognized and cleaved by aspartyl endonuclease (Legumain), such as Ala-Ala, Asn-Ala-Ala.

[0774] In some embodiments, B in the linker unit L of formula (II) or formula (II') of the ADC of the present disclosure is specifically recognized and cleaved by matrix metalloproteinase 2 / 9 (MMP-2 / 9), for example, which is present at high concentrations in tumors, such as Pro-Leu-Gly-Leu-Ala-Gly (PLGLAG), Gly-Pro-Leu-Gly-Ile-Ala-Gly-Gln (GPLGIAGQ), Gly-Pro-Val-Gly-Leu-Ile-Gly-Lys (GPVGLIGK).

[0775] In some embodiments, B as part of the linker unit L of formula (II) or formula (II') of the ADC of the present disclosure is a cleavable peptide containing two or more (e.g., 2-12) continuous or discontinuous amino acids, for example, a dipeptide, tripeptide, tetrapeptide, pentapeptide, hexapeptide, heptapeptide, octapeptide, nonapeptide, decapeptide, undecapeptide, or dodecapeptide unit or consisting thereof. Each amino acid containing peptide unit can be independently selected from natural amino acids or non-natural amino acids and / or be a D- or L-isomer, provided that B contains a cleavable bond that, when cleaved, triggers the release of the KRas mutation inhibitor. In certain embodiments, B consists only of natural amino acids. In other embodiments, B consists ...

Claims

1. Antibody-drug conjugate of formula (X) or a pharmaceutically acceptable salt or solvate thereof: [PL] q -Ab (X) in, Ab represents an antibody or antigen-binding fragment that binds to the target antigen; q represents the number of [PL] units connected to Ab, which is an integer of at least 1 or a non-integer; L represents the linker unit that connects P to Ab; P represents a Ras mutation inhibitor compound.

2. The antibody-drug conjugate according to claim 1, wherein the Ras mutation inhibitor compound is a compound represented by the following formula (I): or a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, prodrug or solvate thereof, wherein: M is selected from N or C-R1; M' is selected from N or C-R1'; R1 and R1' are each independently selected from H, halogen, CN, -C optionally substituted by halogen 1-6 Alkyl and optionally halogen-substituted -OC 1-6 alkyl; R a is selected from H, halogen, CN, -C optionally substituted by halogen 1-6 Alkyl, optionally substituted by halogen -C 2-6 Alkynyl and -OC optionally substituted by halogen or deuterium 1-6 alkyl; X is selected from CH2, N and O; R2 and R2' are each independently selected from H, OH or -C optionally substituted by halogen 1-6 Alkyl; or R2 and R2' attached to non-adjacent ring carbon atoms together form an intracyclic bridge -(CH2) 1-2 - or -CH2=CH2-; or R2 and R2' attached to the same ring carbon atom together with the ring carbon atom to which they are attached form a 4-6 membered spirocycloalkyl or a 4-6 membered spiroheterocycloalkyl containing 1 or 2 heteroatoms selected from N and O; R3 is selected from H, -NH2, -NHC 1-6 Alkyl, -N(C 1-6 Alkyl)2 and -OC 1-6 alkyl; R4 is selected from -CN, halogen, -NO2, -C optionally substituted by halogen 2-6 Alkynyl and optionally halogen-substituted -C 1-6 alkyl; R5 is selected from H, -CN, halogen, -NO2 and halogen substituted -C 1-6 alkyl; R6 is selected from H, halogen, CN, -C 1-6 Alkyl and -C 2-6 Alkynyl, where -C 1-6 Alkyl and -C 2-6 each alkynyl group is independently optionally substituted with halogen; R7, R7', R8 and R8' are each independently selected from H, halogen, CN, -NO2 and -C optionally substituted by halogen 1-6 alkyl; V and W are each independently selected from H, halogen, -C 1-6 Alkyl, OH and NH2; Z is selected from O, N and CH2; R9 and R 10 Each independently selected from H, deuterium, -C 1-6 Alkyl and -(CH2) n -C 3-6 Cycloalkyl, wherein -C 1-6 Alkyl and -C 3- 6 cycloalkyl groups are each independently optionally substituted with deuterium, halogen or -OC 1-6 Alkyl substituted, or R9 and R attached to the same carbon atom 10 Together with the carbon atom to which they are attached, they form C 3-4 Cycloalkyl; R 11 Selected from H, -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl and -(CH2) n -C 3-6 Cycloalkyl, where C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl or C 3-6 The cycloalkyl groups are each independently optionally substituted with deuterium, halogen, CN or -OC 1-6 Alkyl substitution; R 12 Selected from H, halogen, -CN, -OH, -NH2, -NHC 1-6 Alkyl, -N(C 1-6 Alkyl)2, -OC 1-6 Alkyl, -OC 3-6 Cycloalkyl, -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, -(CH2) n -C 3-6 Cycloalkyl and =C(R d )2, where R d are each independently selected from H, halogen and -C 1-6 Alkyl, where each occurrence of C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl or C 3-6 The cycloalkyl groups are each independently optionally substituted with halogen, CN or -OC 1-6 Alkyl substitution; R 13 Selected from H, -C 1-6 Alkyl and -(CH2) n -C 3-6 Cycloalkyl, wherein -C 1-6 Alkyl and -C 3-6 The cycloalkyl groups are each independently optionally substituted with halogen or -OC 1-6 Alkyl substitution; k is an integer selected from 0 or 1 m is an integer selected from 0 to 6; n is an integer selected from 0 to 2; The compound of formula (I) is The NH ring when present in the fragment and / or the OH ring when present on the side chain naphthalene ring or benzothiophene ring are bonded to the linker unit L.

3. The antibody-drug conjugate of claim 2, wherein the structural fragment in formula (I) for Preferably, M is C-R1 and M' is C-R1', wherein R1 is selected from halogen or -C 1-6 Alkyl, R a is H and R1' is halogen, or M is N and M' is C-R1', wherein R a Selected from H, -C 2-6 Alkynyl and -OC 1-6 alkyl, and R1' is halogen, More preferably, M is C-R1 and M' is C-R1', wherein R1 is selected from F or Cl, or -C substituted by halogen 1-3 Alkyl, R a is H, and R1' is F, or M is N and M' is C-R1', wherein R a Selected from H, -C 2-4 Alkynyl and optionally deuterium-substituted -OC 1-3 Alkyl, and R1' is F; and / or The structural fragment in formula (I) In which R3 is selected from -NH2, -NHC 1-6 Alkyl and N(C 1-6 Alkyl) 2; R4 is selected from -CN, halogen, -NO2 and -C substituted by halogen 2-6 Alkynyl; V is selected from H, -C 1-6 Alkyl and halogen, R7 and R7' are each H, or each is halogen, or one of them is H and the other is halogen or halogen substituted C 1-6 alkyl, or C 1-6 alkyl, Preferably, the structural fragment for V is H, R7 and R7' are each H, or one of them is H and the other is halogen, or The structural fragment in formula (I) wherein W is OH; R5 is H or halogen, and R6 is selected from halogen, -C 2-6 Alkynyl and -C 1-6 R8 and R8 'are each H; or R8 and R8 'are each halogen, or one of R8 and R8 'is H, the other is selected from halogen, CN and NO2, or one of R8 and R8 'is H, the other is selected from -C optionally substituted by halogen 1-6 Alkyl, or one of R8 and R8' is selected from H, halogen, -NO2, CN and -C optionally substituted by halogen 1-6 alkyl, the other selected from -C 1-6 alkyl, Preferably, the structural fragment for Wherein R5 is halogen, R6 is selected from -C 2-6 Alkynyl; and / or The structural fragment in formula (I) for wherein R2 and R2' attached to non-adjacent ring carbon atoms together form an intracyclic bridge -CH2-, -CH2CH2- or -CH2=CH2-, preferably and / or Wherein Z is O, and the structural fragment connected to Z is Preferred More preferred 4. The antibody-drug conjugate of claim 2 or 3, wherein R9 and R 10 Both are H, or one or both are deuterium; and / or where R 11 -C 1-6 Alkyl groups wherein the hydrogen atoms are optionally replaced by one or more isotopes of deuterium; and / or where R 12 Selected from -C optionally substituted by halogen 1-6 Alkyl and =C(R d )2, where R d are each independently selected from H and halogen, and m is 1 or 2; and / or where R 13 -C 1-6 Alkyl, preferably -C 1-3 Alkyl, more preferably -CH3.

5. The antibody-drug conjugate according to any one of claims 1 to 4, wherein the structural fragment Selected from 6. The antibody-drug conjugate according to any one of claims 1 to 5, wherein the compound of formula (I) has any of the following sub-formulas: Among them, R1, R1', R a , R3, R4, R5, R6, R7, R7', W, V, R9, R 10 、R 11 、R 12 、R 13 , m are each as defined in claims 1 to 11; preferably When M is C-R1 and M' is C-R1', R1 is selected from F or Cl, or -C substituted by halogen. 1-3 Alkyl, R a is H and R1' is F, or M is N and M' is C-R1', wherein R a Selected from H, -C 2-4 Alkynyl and optionally deuterium-substituted -OC 1-3 Alkyl, and R1' is F; R3 is -NH2, R4 is -CN, V is H, R7 and R7' are each H, or one of them is H and the other is halogen; R5 is halogen, R6 is selected from -C 2-6 Alkynyl, R8 and R8' are each H, and W is OH; R9 and R 10 Both are H, or one or both are deuterium; R 11 -C 1-3 Alkyl groups wherein hydrogen atoms are optionally replaced by one or more isotopes of deuterium; R 12 Selected from -C optionally substituted by halogen 1-3 Alkyl and =C(R d )2, where R d are each independently selected from H and halogen; m is 1 or 2; R 13 -C 1-3 alkyl.

7. The antibody-drug conjugate of any one of claims 1 to 6, wherein the Ras inhibitor compound is selected from: stereoisomers, tautomers, stable isotopic variants, pharmaceutically acceptable salts or solvates thereof.

8. The antibody-drug conjugate of claim 1, wherein the Ras inhibitor compound has any of the following sub-formulas: or a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof, wherein M, M', X, Z, V, W, R1, R1', R3, R4, R5, R6, R7, R7', R8, R8', R9, R 10 、R 11 、R 12 、R 13 , k, m, n, structural fragments As defined in any one of claims 2 to 6 respectively; Y is selected from -CH2-, -CH2CH2-, -CH2OCH2- or is absent; R 14 and R 15 Each independently selected from H and -C 1-6 alkyl; Preferably, exist In the fragment, when M is C-R1, R1 is selected from halogen (preferably F or Cl) or -C 1-6 Alkyl (preferably halogen-substituted -C 1-3 alkyl, more preferably -CF3), and M' is C-R1' and R1' is halogen (preferably F); or when M is N, M' is C-R1' and R1' is halogen (preferably F); exist In the fragment, both M are C-R1, or one M is N and the other M is C-R1, and R1 is independently selected from H, halogen (preferably F) and halogen-substituted -C 1-6 Alkyl (preferably one or more F substituted -C 1-3 Alkyl, more preferably -CF3), preferably the M at the ortho position of R3 is selected from N and C-R1 (preferably C-halogen, more preferably CF), and the M at the ortho position of R6 is C-R1 (preferably C-halogen substituted -C 1-3 Alkyl, more preferably -C-CF3); R3 is selected from -NH2, -NHC 1-6 Alkyl and N(C 1-6 Alkyl)2 (preferably selected from -NH2, -NHC 1-3 Alkyl and N(C 1-3 alkyl) 2, more preferably -NH2); R5 is H or halogen (preferably F); R6 is selected from halogen (preferably Cl) and -C 1-6 Alkyl (preferably -C 1-3 alkyl); exist In the fragment, R3 is selected from -NH2, -NHC 1-6 Alkyl and N(C 1-6 Alkyl)2 (preferably selected from -NH2, -NHC 1-3 Alkyl and N(C 1-3 Alkyl) 2, more preferably -NH2); R4 is selected from -CN, halogen, -NO2 and -C substituted by halogen 2-6 Alkynyl; V is selected from H, -C 1-6 Alkyl (preferably -C 1-3 R7 and R7' are each H, or each halogen, or one of them is H and the other is halogen or halogen-substituted C 1-6 Alkyl (preferably halogen-substituted -C 1-3 alkyl), or one of which is halogen and the other is halogen-substituted C 1-6 Alkyl (preferably halogen-substituted -C 1-3 Alkyl), wherein the halogen is preferably F; Preferably R3 is -NH2, R4 is -CN, V is H, R7 and R7' are each H, or one of them is H and the other is halogen (preferably F); exist In the fragment, R5 is H or halogen (preferably F); R6 is selected from halogen, -C 2-6 Alkynyl (preferably -C 2-4 Alkynyl) and -C 1- 6 alkyl (preferably -C 1-3 R8 and R8' are each H, or R8 and R8' are each halogen, or one of R8 and R8' is H, the other is selected from halogen, CN and NO2, or one of R8 and R8' is H, the other is selected from -C optionally substituted by halogen 1-6 Alkyl, or one of R8 and R8' is selected from H, halogen, -NO2, CN and -C optionally substituted by halogen 1-6 alkyl, the other selected from -C 1-6 Alkyl; W is OH; Preferably R5 is halogen (preferably F), R6 is selected from -C 2-6 Alkynyl (preferably -C 2-4 alkynyl), R8 and R8' are each H, and W is OH; X is selected from O and NH; Y is selected from -CH2CH2- and absent; Z is selected from O; R9 and R 10 Both are H; or one or both are deuterium; R 11 -C 1-6 Alkyl (preferably -C 1-3 alkyl), wherein the hydrogen atoms are optionally replaced by one or more deuterium; R 12 Selected from -C optionally substituted by halogen 1-6 Alkyl (preferably -C 1-3 alkyl) and =C(R d )2, where R d are each independently selected from H and halogen (preferably F); m is 0, 1 or 2 (preferably 1 or 2); n and k are 0 or 1 (preferably 0); R 13 -C 1-6 Alkyl (preferably -C 1-3 alkyl, more preferably -CH3); R 14 and R 15 H or -C 1-3 Alkyl (preferably -CH3); 9. The antibody-drug conjugate of claim 1, wherein the Ras inhibitor compound is selected from the group consisting of: or a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt, or solvate thereof.

10. The antibody-drug conjugate of any one of claims 1 to 9, wherein the linker unit L has: -A(S)-BG-(II-3) or -BG-(II-5) in: A is selected from Preferred wherein one end marked with * is connected to drug P, and N or O marked with * is an atom from drug P; Y1 is independently NH or O; R 16 H; R 17 is H; S is selected from the group consisting of glucose, galactose, mannose, glucosamine, galactosamine, mannosamine, galacturonic acid, glucuronic acid, mannuronic acid, acetylgalactose, acetylglucosamine, acetylmannose, acetylgalactosamine, acetylglucosamine, acetylmannosamine, glucosamine, galactosamine, mannosamine; lactose, maltose or their respective amino derivatives, acetylamino derivatives, uronic acid derivatives, aminouronic acid derivatives, acetylaminouronic acid derivatives; preferably, S is selected from the group consisting of D-glucose Sugar, D-galactose, D-mannose, D-galacturonic acid, D-glucuronic acid, D-mannuronic acid, 2-acetylamino-2-deoxy-D-galactose, 2-acetylamino-2-deoxy-D-glucose, 2-acetylamino-2-deoxy-D-mannose, 2-acetylamino-2-deoxy-D-galacturonic acid, 2-acetylamino-2-deoxy-D-glucuronic acid, 2-acetylamino-2-deoxy-D-mannuronic acid; D-lactose, D-maltose, D-sucrose or their respective 2-acetylamino and / or 6-carboxyl derivatives; B is the formula (B-2): -(AA) d -,in d is an integer from 2 to 12, preferably an integer from 2 to 4; AA is an amino acid selected from the group consisting of alanine, glycine, asparagine, valine, phenylalanine, citrulline, and glutamic acid; preferably -(AA) d - is a polypeptide from the following C-terminus to the N-terminus: Ala-Ala, Cit-Val, Ala-Val, Glu-Gly, Asn-Ala-Ala, Cit-Ala-Glu, Gly-Phe-Gly-Gly; wherein the N-terminus of the peptide is linked to the G moiety and the C-terminus of the peptide is linked to the self-immolative linker A; G is selected from The * end indicates the connection point with the antibody Ab, End represents the connection point with B; Specifically, the linker unit L has the formula (II-3-1): And the maleimide portion thereof is The corresponding general formula fragment, wherein -O- in -OS is O in the solubilizing sugar unit that forms a glycosidic bond with the benzene ring, or Specifically, the linker unit L has the following formula when the solubilizing sugar unit S is absent: And the maleimide portion thereof is or The linker unit L has the following formula: And the maleimide portion thereof is The corresponding general formula fragment of .

11. The antibody-drug conjugate of claim 10, wherein the solubilizing sugar unit in the linker unit is selected from the group consisting of: in, The wavy line shows the connection between the solubilizing sugar unit and the self-immolative linker A. The connected O is the O in the solubilizing sugar unit that forms a glycosidic bond with the benzene ring. The dotted line of the 1-position bond of the solubilizing sugar unit indicates that the configuration of the glycosidic bond can be α-type, β-type or a mixture thereof.

12. The antibody-drug conjugate of any one of claims 10-11, wherein B in the Linker unit is selected from the following polypeptides: The left end carbonyl group is connected to the self-degradable linker A or drug P, and the right end N is connected to the G part.

13. The antibody-drug conjugate of claim 1, wherein the [PL] conjugated fragment has any of the following general formulas: And the maleimide portion thereof is The corresponding general formula fragment of or its stereoisomers, tautomers, stable isotopic variants, pharmaceutically acceptable salts or solvates, wherein M, M', R a , R3, R4, R5, R6, R7, R7', R8, R8', R9, R 10 、R 11 、R 12 、R 13 , W, V, m, and each structural fragment have the meanings defined in claims 2-9, respectively, and A, S, B, G, AA, and d in the linker unit L have the meanings defined in claims 10-12, respectively; The wavy line indicates the connection point with Ab; the atom marked with * is the site in the drug P unit that is connected to the linker unit.

14. The antibody-drug conjugate of claim 1, having any general formula selected from the group consisting of: And the maleimide portion thereof is The corresponding conjugates or a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof, wherein: M, M', R in drug P unit a , R3, R4, R5, R6, R7, R7', R8, R8', R9, R 10 、R 11 、R 12 、R 13 , W, V, m, and each structural fragment have the meanings defined in claims 2-9, respectively, and A, S, B, G, AA, and d in the linker unit L have the meanings defined in claims 10-12, respectively; The atom marked with * is the site where the drug P unit is connected to the linker unit; q is an integer or non-integer from 1 to 10, or a range consisting of any two values ​​between 1 and 10; preferably, q is an average DAR value from 1 to 8, or a range consisting of any two values ​​between 1 and 8.

15. The antibody-drug conjugate of any one of claims 1 to 9, wherein the linker unit L has the structure of formula (II'-3), -JBG-(II'-3) in, J is selected from -J1-, -J2-, -J1-J2-, -J2-J1-, wherein -J1-Selected Wherein, * indicates the connection point with drug P or -J2-, and the wavy line indicates the connection point with cleavable linker B; -J2-Selected Wherein, * indicates the connection point with drug P or -J1-, and the wavy line indicates the connection point with cleavable linker B; Each occurrence of X' is independently selected from CH or N; Each occurrence of Y1' is independently selected from -NH-, -NC 1-6 Alkyl- or -O-; Each occurrence of R 16 ' are each independently selected from H and -C 1-6 Alkyl, preferably H or C 1-3 Alkyl, more preferably H; Each occurrence of R 17 ' are each independently selected from H, -NO2, -NH2, -CF3 or a fragment of formula (A1): Each occurrence of R 18 Each independently selected from -C 1-6 Alkyl, preferably C 1-3 Alkyl, more preferably methyl; T is selected from -C(R a )2-、-O-、-NR a - or does not exist; T1 is selected from -C(R a )2-、-O-、-NR a -; T2 selected from -CR a -and-N-; T3 is a peptide consisting of 2 to 6 amino acids connected to N at the carboxyl terminus; Q is selected from -CO-, -O-, -NR a - or does not exist; U is selected from glycosyl or its derivatives, and contains Hydrophilic peptides; Subunit selected from natural amino acid residues and unnatural amino acid residues; R a Each independently selected from H or -C 1-6 Alkyl, preferably H or -CH3; p is an integer from 0 to 4, such as 0-2, 1-4, 1-2, 2-4; t is an integer from 0 to 10, for example, 0-8, 0-6, 0-4, 0-2, 1-10, 2-10, 2-8, 2-6, 2-4, 6-10, 8-10; B is the formula (B-2): -(AA) d -, wherein d is an integer from 2 to 12, preferably an integer from 2 to 4, preferably -(AA) d - is a polypeptide from C-terminus to N-terminus of the following: Ala-Ala, Cit-Val, Ala-Val, Glu-Gly, Asn-Ala-Ala, Cit-Ala-Glu, Gly-Phe-Gly-Gly, wherein the N-terminus of the peptide is linked to the G portion of the Linker unit and the C-terminus of the peptide is linked to J; G is Where G1 is the atom in Ab responsible for connecting to L, G3' is -C 1-10 Alkylene-, preferably -C 1-5 Alkylene-, Bu does not exist or is of formula (A) The structural fragment, wherein T0 is -C 1-6 Alkylene-, preferably -C 1-2 Alkylene-, more preferably methylene, p, Q, U are each as defined above.

16. The antibody-drug conjugate of claim 15, wherein J is selected from The * indicates the connection point with the drug P, and the wavy line indicates the connection point with the rest of the linker unit; Each occurrence of X' is independently selected from -N- or -CH-; Each occurrence of Y1' is independently selected from -NH- or -O-; wherein Y1' attached to the six-membered aromatic ring is attached to the para or ortho position of the other attachment point of the ring; Each occurrence of R 16 ' are each independently selected from H or -C 1-3 Alkyl, preferably H; Each occurrence of R 17 ' are each independently selected from H or a fragment of formula (A1) Each occurrence of R 18 Each independently selected from -C 1-3 Alkyl, preferably methyl; Each occurrence of T is selected from -O-, -NR a - or does not exist; Each occurrence of T1 is selected from -CH2-, -O-, -NCH3-; Each occurrence of T2 is selected as -N-; Each occurrence of T3 is a 2-4 peptide linked to N at its carboxyl terminus, selected from Ala-Ala, Cit-Val, Ala-Val, Glu-Gly, Asn-Ala-Ala, Cit-Ala-Glu, Gly-Phe-Gly-Gly; Each occurrence of Q does not exist or is -NR a -or-O-; Each occurrence of U is selected from Hydrophilic peptides such as or a saccharide (preferably a monosaccharide or disaccharide or its uronic acid) or a derivative thereof, preferably a hydrophilic peptide, wherein t is 2-14, such as 4-14, 6-12, preferably 10; each occurrence of p is 0-4, preferably 0-2; For example, the self-immolative linker J is selected from:

17. The antibody-drug conjugate of claim 15 or 16, wherein G is Where T0 is -C 1-4 Alkylene-(e.g. -C 1-2 Alkylene-, such as methylene), T is absent or is -C(R a )2-, p is 0-4 (e.g., 0-2) and Q is selected from -O- or -NR a -, and U is selected from uronic acid (preferably uronic acid of monosaccharide or disaccharide) or its derivatives and is linked to a carbonyl group, Hydrophilic peptides such as or where T0 is -C 1-4 Alkylene-(e.g. -C 1-2 Alkylene-, such as methylene), T is absent or is -C(R a )2-, p is 0-4 (e.g., 0-2) and Q is -CO-, U is selected from amino sugars (preferably amino monosaccharides or amino disaccharides) or derivatives thereof and is linked to an amino group, or hydrophilic peptides such as or where T0 is -C 1-4 Alkylene-(e.g. -C 1-2 Alkylene-, such as methylene), T is selected from -C(R a )2-、-O-、-NR a - or does not exist, Q does not exist, p is 0-4 (e.g., 0-2, 1-2), and U is selected from a glycosyl (preferably a monosaccharide or disaccharide) or a derivative thereof and is linked by a glycosidic bond, or an amino sugar (preferably an aminomonosaccharide or an aminodisaccharide) or a derivative thereof and is linked by an amino group; wherein t is an integer from 1 to 20, for example, 2-10, 2-8, 2-6, 2-4, 4-10, 4-14, 6-10, 6-12, 8-10, 8-14.

18. The antibody-drug conjugate of any one of claims 15 to 17, wherein B is as defined in claim 12, and / or a subunit of a hydrophilic peptide a residue corresponding to arginine, serine, threonine, tyrosine, cysteine, aspartic acid, asparagine, glutamic acid, glutamine, lysine, histidine, glycine, tryptophan, ornithine, citrulline, sarcosine, or R, R′, R″ are each independently additionally a group containing a carboxyl group, a sulfonic acid group, a phosphate group, an amino group, an amide group, a quaternary ammonium group, a sulfhydryl group and / or a hydroxyl group; Preferably, the hydrophilic peptide comprises 4-14 units of polysarcosine, polyarginine, polyglycine, preferably 6-12 units of polysarcosine.

19. The antibody-drug conjugate of any one of claims 1-9 and 15-18, wherein the Linker unit has the formula: wherein each variable is as defined in any one of claims 15-18; For example, the linker unit L is selected from the structural fragments shown in Table 7 of the specification.

20. The antibody-drug conjugate of claim 1, wherein the [PL] conjugated fragment has any of the following general formulas: wherein the -JBG- fragment is as defined in any one of claims 15 to 19, preferably as defined in claim 19, and the drug P unit is a Ras inhibitor compound as defined in any one of claims 1 to 9, preferably a Ras inhibitor compound as defined in claim 7 or 9, or a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof; Preferably, the [PL] conjugated fragment is the conjugated fragment shown in Table 8-1 of the specification.

21. The antibody-drug conjugate of claim 1 , having a general formula selected from the group consisting of: wherein the -JBG- fragment is as defined in any one of claims 15 to 19, preferably as defined in claim 19; The drug P unit is a Ras inhibitor compound as defined in any one of claims 1 to 9, preferably a Ras inhibitor compound as defined in claim 7 or 9, or a stereoisomer, tautomer, stable isotope variant, pharmaceutically acceptable salt or solvate thereof; q is 1 to 10, or the average DAR value of any two values ​​between 1 and 10, for example, about 1-10, 1-8, 2-8, 3-10, 3-8, 4-10, 4-8, 6-8 or 6-10.

22. The drug-antibody conjugate of any one of claims 1 to 21, wherein Ab is an antibody or antigen-binding fragment thereof that binds to a tumor-specific antigen or tumor-associated antigen, wherein the tumor-specific antigen or tumor-associated antigen is selected from the group consisting of: HER2, Her3, HER1 (ErbB1), HER4 (ErbB4), TROP2, Nectin-4, tissue factor, PD-L1, PD-1, PD-L1 / PD-L2, MET, CLDN18.2, KI T, CTLA-4, RPR1, adrenergic A2 receptor (EphA2), folate receptor (FRa), mesothelin, endothelin receptor, GCPII, IL-13Ra, BCMA, GD2, CLL-1, CA-IX, MUC1, 5T4, AOC3, ALK, AXL, C242, CA-125, CCL11, CCR5, CD2, CD3, CD4, CDS, CD15, CA15-3, CD16, CD18, CD1 9. CD20, CD21, CD22, CD25, CD30, CD32, CD33, CD37, CD38, CD44, CD52, CD56, CD64, CD66e, CD70, CD72, CD 74. CD79a, CD79b, CD123, CD138, CD142, CD174, CD276, CDH3, CCD79b, CLDN9 / CLDN6, CA19-9, DPEP3, AGS- 16. IGF1R, IGF2R, VEGFR1, VEGFR2, VEGFR3, PDGFR-α, PDGFR-β, EGFR, EGFRvIII, ENPP3, FcRH5, FRα, KAA G1, LIV-1, Mesothelin, cMet, ROR1, SLTRK6, TF, BMPR1B, E16, TOP1, STEAP1, 0772P, MUC16, Napi3b, Sema 5b. PSCAhIg, ETBR, RNF124, prostate cancer-associated gene 1, TrpM4, teratoma-derived growth factor 1, C3DR, FcRH2, NCA, MDP, IL20R-α, Brevican, EphB2R, ASLG659, prostate stem cell antigen precursor, GEDA, BAFF-R, CXCR5, HLA-DOB, P2X5, LY64, FcRH1, IRTA2, TENB2, integrin α5β6, integrin α4β7, FGF2, FGFR1, FGFR2, FGFR3, FGFR4, PSMA, somatostatin receptor, RANK, SLAMF7, ITGB6, CEACAM5, CA9, EGFRvl 11, IL2RA, AXL receptor tyrosine kinase, TGF-βR, TNFRSF8, cancer / testis-associated antigen, CLEC14A, GRP78, stem cell-specific antigen, ASG-5, PRR4, GUCY2C, SLC39A6, TPBG, tumor-associated antigen CA242, FOLR1, GPNMB, HAVCR1, prostate tumor target Mindin, VTCN1, PTK7 protein tyrosine kinase 7, macrophage stimulating 1 receptor, TACSTD2, CA6, DLL3, DLL4, EpCAM, FAP, DKK-1, Endoglin, VCAM1, GPC3, DR5, ASCT2, B7H1, B7H3, B7H4, or any combination thereof; Preferably, the tumor-specific antigen or tumor-associated antigen is selected from AXL, B7H1, B7H3, B7H4, BCMA, CD16, CD19, CD22, CD25, CD30, CD32, CD33, CD44, CD64, CD70, CD74, CD79, CD138, CD142, CD276, CDH3, CEACAM5, Claudin 18.2, CLDN9 / CLDN6, DPEP3, EGFR, ENPP3, EphA, FcRH5, FOLR1, FRα, GCPII, HER2, HER3, KAAG1, KIT, LIV-1, Mesothelin, cMet, MUC1, Nectin-4, PD-L1, PD-L1 / PD-L2, PSMA, ROR1, RPR1, TF, TOP1, TROP2, or any combination thereof; More preferably, the tumor-specific antigen or tumor-associated antigen is selected from HER2, Claudin 18.2, EGFR, TROP2, Nectin-4, or a combination of EGFR and Met.

23. The drug-antibody conjugate of any one of claims 1 to 22, wherein: The Ab comprises three heavy chain complementary determining regions (HCDRs) and three light chain complementary determining regions (LCDRs) that specifically bind to HER2, wherein: according to the Kabat definition, HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 6, HCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 7, HCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 8, LCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 3, LCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 4, and LCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 5; Or the Ab comprises three heavy chain complementary determining regions (HCDRs) and three light chain complementary determining regions (LCDRs) that specifically bind to Trop2, wherein: according to the Kabat definition, HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 18, HCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 19, HCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 20, LCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 15, LCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 16, and LCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 17; Or the Ab comprises three heavy chain complementary determining regions (HCDRs) and three light chain complementary determining regions (LCDRs) that specifically bind to Claudin18.2, wherein: according to the Kabat definition, HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 29, HCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 30, HCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 31, LCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 26, LCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 27, and LCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 28; Or the Ab comprises three heavy chain complementary determining regions (HCDRs) and three light chain complementary determining regions (LCDRs) that specifically bind to EGFR, wherein: according to the Kabat definition, HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 40, HCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 41, HCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 42, LCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 37, LCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 38, and LCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 39; Or the Ab comprises three heavy chain complementary determining regions (HCDRs) and three light chain complementary determining regions (LCDRs) that specifically bind to Nectin4, wherein: according to the Kabat definition, HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 72, HCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 73, HCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 74, LCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 69, LCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 70, and LCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 71, Or the Ab comprises a first set of complementarity determining regions that specifically bind to EGFR and a second set of complementarity determining regions that specifically bind to MET, wherein, The first set of complementarity determining regions comprises three heavy chain complementarity determining regions (HCDRs) and three light chain complementarity determining regions (LCDRs), wherein: according to the Kabat definition, HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 49, HCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 50, HCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 51, LCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 52, LCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 53, and LCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 54, The second set of complementarity determining regions comprises three heavy chain complementarity determining regions (HCDRs) and three light chain complementarity determining regions (LCDRs), wherein: according to the Kabat definition, HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 55, HCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 56, HCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 57, LCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 58, LCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 59, and LCDR3 comprises or consists of the amino acid sequence of SEQ ID NO:

60.

24. The drug-antibody conjugate of any one of claims 1 to 22, wherein Ab comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 10, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 9; or wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 22, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 21; or wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 33, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 32; or wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 44, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 43; or wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 76, and wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO: 75; or The Ab comprises a first heavy chain variable region and a first light chain variable region and a second heavy chain variable region and a second light chain variable region, wherein the first heavy chain variable region and the first light chain variable region comprise the amino acid sequences of SEQ ID NO: 61 and SEQ ID NO: 62, respectively, and wherein the second heavy chain variable region and the second light chain variable region comprise the amino acid sequences of SEQ ID NO: 63 and SEQ ID NO: 64, respectively.

25. The drug-antibody conjugate of any one of claims 1 to 22, wherein Ab comprises: (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 2, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 1; or (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 14, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 13; or (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 25, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 24; or (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 36, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 35; or (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 76, and (b) a light chain comprising the amino acid sequence of SEQ ID NO: 75, or (a) a first heavy chain comprising the amino acid sequence of SEQ ID NO: 45, (b) a first light chain comprising the amino acid sequence of SEQ ID NO: 46, (c) a second heavy chain comprising the amino acid sequence of SEQ ID NO: 47, and (d) a second light chain comprising the amino acid sequence of SEQ ID NO:

48.

26. The drug-antibody conjugate according to any one of claims 1 to 22, wherein Ab is selected from trastuzumab or an antibody fragment thereof, or other anti-human HER2 antibodies that recognize the same epitope or competitively bind to human HER2, preferably trastuzumab; or wherein Ab is selected from certolizumab pegol or its antibody fragment, or other anti-human TROP2 antibodies that recognize the same epitope or competitively bind to human TROP2, preferably certolizumab pegol; or wherein Ab is selected from zotuximab or its antibody fragment, or other anti-human Claudin18.2 antibodies that recognize the same epitope or competitively bind to human Claudin18.2, preferably zotuximab; or wherein Ab is selected from cetuximab or its antibody fragment, or other anti-human EGFR antibodies that recognize the same epitope or competitively bind to human EGFR, preferably cetuximab; or wherein the Ab is selected from Enfortumab or its antibody fragment, or other anti-human Nectin4 antibodies that recognize the same epitope or competitively bind to human Nectin4, preferably Enfortumab; or The Ab is selected from ervantumab or its antibody fragment, or other anti-human EGFR and MET antibodies that recognize the same epitope or competitively bind to human EGFR and MET, preferably ervantumab.

27. A drug-antibody conjugate selected from the group consisting of: or a stereoisomer, tautomer, stable isotopic variant, pharmaceutically acceptable salt or solvate thereof; Preferred are the drug-antibody conjugates of Examples 1-94 or stereoisomers, tautomers, stable isotope variants, pharmaceutically acceptable salts or solvates thereof.

28. A pharmaceutical composition comprising the drug-antibody conjugate according to any one of claims 1 to 27 and one or more pharmaceutically acceptable excipients.

29. The pharmaceutical composition according to claim 28, which is administered intravenously, intratumorally, subcutaneously, intramuscularly, orally, intranasally, intrathecally, transdermally or topically, preferably intravenously, intraperitoneally, subcutaneously or intramuscularly.

30. The antibody-drug conjugate of any one of claims 1 to 27 or the pharmaceutical composition of claim 28 or 29, for use in treating or preventing diseases mediated by Ras mutant proteins, such as KRas mutant proteins, particularly KRas G12D mutant proteins, preferably for treating or preventing hyperproliferative diseases, more preferably for treating or preventing tumors.

31. Use of the antibody-drug conjugate of any one of claims 1 to 27 or the pharmaceutical composition of claim 28 or 29 in treating or preventing diseases mediated by Ras mutant proteins, such as KRas mutant proteins, particularly KRas G12D mutant proteins, preferably hyperproliferative diseases, more preferably tumors.

32. Use of the antibody-drug conjugate of any one of claims 1 to 27 or the pharmaceutical composition of claim 28 or 29 in the preparation of a medicament for treating or preventing a disease mediated by a Ras mutant protein, such as a KRas mutant protein, particularly a KRas G12D mutant protein, preferably a hyperproliferative disease, more preferably a tumor.

33. A method for treating or preventing a disease mediated by a Ras mutant protein, such as a KRas mutant protein, particularly a KRas G12D mutant protein, preferably a hyperproliferative disease, more preferably a tumor, comprising administering to a human or animal the antibody-drug conjugate of any one of claims 1 to 27 or the pharmaceutical composition of claim 28 or 29.

34. The antibody-drug conjugate of claim 30, or the pharmaceutical composition of claim 28 or 29, the use of claim 31 or 32, or the method of claim 33, wherein the hyperproliferative disease or tumor includes solid tumors and blood-borne tumors, and all precancerous and cancerous cells and tissues selected from the group consisting of lung adenocarcinoma, lung cancer (including squamous cell carcinoma of the lung and non-small cell lung cancer, small cell lung cancer), bone cancer, pancreatic cancer, pancreatic ductal adenocarcinoma, skin cancer, head and neck cancer (including head and neck squamous cell carcinoma), melanoma (including skin or intraocular melanoma), squamous cell carcinoma, anal region cancer, testicular cancer, urethra cancer, ureteral cancer, penile cancer, prostate cancer (including hormone-refractory prostate cancer), bladder cancer, uterine cancer, ovarian cancer, epithelial ovarian cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, stomach cancer, gastric adenocarcinoma, colon cancer, rectal cancer, colorectal cancer, liver cancer, breast cancer (including metastatic breast cancer, triple-negative breast cancer (TNBC)), esophageal cancer, small intestine cancer, lip cancer, laryngeal cancer, nasopharyngeal cancer, oral cancer, salivary gland cancer, peritoneal cancer, gastrointestinal stromal tumors, gastroesophageal junction (GEJ) J) carcinoma, mesothelioma, biliary tract cancer, hepatoma, seminoma, soft tissue sarcoma, osteosarcoma, urothelial cancer, sweat gland cancer, endocrine system cancer, thyroid cancer, medullary thyroid cancer, follicular thyroid cancer, papillary thyroid cancer, parathyroid cancer, kidney cancer, renal parenchymal cancer, renal cell carcinoma, renal pelvis cancer, adrenal gland cancer, brain cancer such as glioblastoma, astrocytoma, meningioma, medulloblastoma, peripheral neuroectodermal tumor, glioblastoma (including glioblastoma multiforme), neuroblastoma; chronic or acute leukemia, Hodgkin's disease, lymphoma (including lymphocytic lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, Burkitt lymphoma, adult T-cell lymphoma, diffuse large-scale lymphoma (DLBCL), primary CNS lymphoma), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CLL) and lymphocytic carcinoma, acute myeloid leukemia (AML), myeloid leukemia (chronic myeloid leukemia (CML), central nervous system tumors (CNS), spinal tumors, brainstem gliomas or pituitary adenomas.

35. The antibody-drug conjugate or pharmaceutical composition, use, or method of claim 34, wherein the tumor is selected from the group consisting of lung cancer, lung adenocarcinoma, colon cancer, rectal cancer, pancreatic cancer, endometrial cancer, bile duct cancer, leukemia, and ovarian cancer.

36. A compound fragment having any of the general formulae (II'-3'-1) to (II'-3'-12) and (II'-4'-1) as defined in claim 19, preferably a structural fragment shown in Table 7 of the specification.

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