Dual payload antibody drug conjugates

Abstract

The present invention relates to dual payload antibody drug conjugates comprising at least one topoisomerase inhibitor and at least one DNA damage response (DDR) inhibitor, pharmaceutical compositions thereof, and uses of the antibody drug conjugates and compositions thereof in the treatment of diseases and conditions, including proliferative diseases.

Description

[0001] Cross-references to related applications This application claims and enjoys priority to U.S. Provisional Application No. 63 / 590,365, filed October 13, 2023, the entire contents of which are incorporated herein by reference for all purposes.

[0002] Invention Field This invention relates to dual-load antibody-drug conjugates comprising at least one topoisomerase inhibitor and at least one DNA damage response (DDR) inhibitor, pharmaceutical compositions thereof, and the use of said antibody-drug conjugates and compositions thereof in the treatment of diseases and conditions, including proliferative diseases. Background of the Invention Topoisomerase 1 (Top1) is an important enzyme that helps unwind supercoiled double-stranded DNA. Under normal conditions, Top1 attacks the phosphodiester bonds on the DNA strand via tyrosine residues to form a covalent bond with the 3'-phosphate terminus, simultaneously releasing the 5'-hydroxyl terminus, thus "cutting" the DNA. This DNA break is transient; after unwinding, the 5'-hydroxyl terminus immediately attacks the 3'-phosphate terminus, causing the DNA to rejoin and releasing Top1.

[0004] In cancer cells, this breakage can trigger apoptosis. Topoisomerase I (Top1) inhibitors are a class of chemotherapeutic drugs that target and capture the Top1-DNA covalent complex (Top1cc), inhibiting the DNA rejoining step and thus causing single-strand breaks. Furthermore, the captured Top1cc is toxic and can cause double-strand breaks during replication. Known Top1 inhibitors include camptothecin and its derivatives, such as topotecan, irinotecan, rubitecan, exatecan, gly-exatecan, belotecan, and 7-ethyl-10-hydroxycamptothecin (SN-38). Top1 inhibitors interfere with DNA replication in cancer cells by stabilizing Top1cc, ultimately leading to apoptosis.

[0005] However, cancer cells typically activate DNA response pathways rapidly in response to topoisomerase I (Top1) inhibition to repair and mitigate damage. Cancer cells have evolved the ability to respond to both single-strand breaks (SSBs, which are more common) and double-strand breaks (DSBs, which are more lethal). These DNA damage response (DDR) pathways limit the toxicity of Top1 inhibition by repairing the damage and arresting the cell cycle during the repair process. For example, the presence of Top1cc can activate the ubiquitin-proteasome pathway to degrade Top1, resulting in a 3'-phosphate terminus linked to a small peptide and a free 5'-hydroxy terminus. Once such a single-strand break is detected, the enzyme poly(ADP-ribose) polymerase 1 (PARP1) binds to the damage site and activates downstream repair mechanisms to mitigate the toxic effects of the Top1cc complex in the single-strand break repair mechanism. et al. Biomedicine& Pharmacotherapy 2020, 125, 109875). If single-strand breaks are not repaired in time, they can lead to double-strand breaks. Typically, camptothecin-induced DNA damage is repaired via the homologous recombination (HR) pathway; this HR pathway is most efficient in the S and G2 phases, while camptothecin-induced DNA damage is S-phase specific. Upon detection of a double-strand break, repair mechanisms are triggered by ataxia-telangiectasia and Rad3-associated (ATR) kinases, as well as ataxia-telangiectasia mutant (ATM) kinases and their downstream targets (checkpoint kinase 1 (CHK1) and checkpoint kinase 2 (CHK2), respectively), thereby repairing the damage (Mei). et al. Biomedicine&Pharmacotherapy 2020, 125, 109875).

[0006] Cancers with defective DNA damage response (DDR) pathways often compensate by over-reliance on another DDR pathway. For example, PARP inhibitors are effective against cancers carrying specific DDR pathways. BRCA1 or BRCA2 ( BRCA1 / 2 Cancers with mutated genes (i.e., cancers with homologous recombination deficiency (HRD)) show significant therapeutic effects (Pilie). et al. Nat Rev Clin Oncol (2019, 16, 81-104). To treat cancer, this dependence can be exploited by specifically administering DNA damage response (DDR) inhibitors that target this particular pathway.

[0007] Synthetic lethality refers to the loss of function resulting from the simultaneous perturbation of two genes, while perturbation of only one gene is tolerable. One approach to treating cancer using synthetic lethality is the simultaneous administration of both a topoisomerase I (Top1) inhibitor and a DNA damage response (DDR) inhibitor. For example, studies have shown that inhibiting the catalytic activity of PARP1 can enhance the cytotoxicity of Top1 inhibitors (Bowman's Law). et al. British Journal of Cancer2001, 84, 106-112; Kinneer et al. Clin Cancer Res 2023, 29 (6): 1086–1101). Nevertheless, combination therapy with systemic Top1 inhibitors and PARP1 inhibitors has not yet been clinically realized due to dose-limiting toxicities. Myelosuppression is a serious toxic side effect that has severely limited dose escalation of both PARP1 inhibitors and chemotherapy drugs in multiple clinical trials (Thomas A. and Pommier Y.). Clin Cancer Res 2019, 25, 6581-6589. One example is that when the PARP1 inhibitor veliparib is used in combination with the Top1 inhibitor topotecan, both drugs require dose reduction due to myelosuppression (the maximum tolerated dose (MTD) of veliparib and topotecan in combination is only 3% and 40% of their respective single-agent MTDs, respectively (Thomas A. and Pommier Y.). Clin Cancer Res 2019, 25, 6581-6589; Kummar et al. Clin Cancer Res 2012, 71, 5626).

[0008] Therefore, there is a need for drug delivery platforms that can safely administer Top1 inhibitors and DNA response inhibitors to leverage their synergistic effects and simultaneously improve the therapeutic efficacy and therapeutic window of these two individual drugs.

[0009] Invention Abstract This invention describes antibody-drug conjugates (ADCs) comprising an antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment is covalently linked via a first linker to at least one topoisomerase inhibitor, and the antibody or antigen-binding fragment is further covalently linked via a second linker to at least one DNA damage response (DDR) inhibitor. In some embodiments, the synergistic ADCs disclosed herein exhibit superior efficacy and therapeutic index compared to the administration of either a topoisomerase inhibitor or a DDR inhibitor alone.

[0010] In some implementations, the ADC is structured as shown in equation (I): Formula (I) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof; Where Ab is the antibody or its antigen-binding fragment; L a It is the first connector; L b It is the second connector; Each DDRi is an independent residue of a DDR inhibitor; Each TOPOi is an independent residue of a topoisomerase inhibitor; The subscript n is an integer selected from 1 to 10; and The subscript m is an integer selected from 1 to 10.

[0011] On the other hand, the present invention provides a pharmaceutical composition comprising the antibody-drug conjugate (ADC) described herein. In a further aspect, the present invention provides a kit comprising the ADC described herein or a pharmaceutical composition thereof.

[0012] On the other hand, the present invention provides a method for treating a disease, symptom, or condition (including cancer and proliferative diseases), the method comprising administering the ADC or a pharmaceutical composition thereof described herein to a subject in need. In some embodiments, the disease, symptom, or condition is cancer.

[0013] In a further aspect, the present invention provides a connector payload comprising a DDR inhibitor. In other aspects, the present invention also provides a DDR inhibitor payload.

[0014] Brief description of the attached figures Figure 1 A comparison of the Kabat and Chothia numbering systems for CDR-H1 is provided. Adapted from Martin ACR (2010). Protein Sequence and Structure Analysis of Antibody Variable Domains. In R. Kontermann & S. Dübel (Eds.), Antibody Engineering vol. 2 (pp. 33-51). Springer-Verlag, Berlin Heidelberg.

[0015] Figure 2A It is the LCMS of the light chain that is coupled with DDRiLP108 to form ADC 5 in Table 1.

[0016] Figure 2B It is the LCMS of the heavy chain that is coupled with TOPOiLP201 to form ADC 5 in Table 1.

[0017] Figure 3A It is the LCMS of the light chain that is coupled with DDRiLP103 to form ADC 8 in Table 1.

[0018] Figure 3BIt is the LCMS of the heavy chain that is coupled with TOPOiLP201 to form ADC 8 in Table 1.

[0019] Figure 4A The MC38-h TF activities of the dual conjugates ADC7 and ADC8, as well as the comparative control ADC15, are shown.

[0020] Figure 4B The MC38-h TF activities of the dual conjugates ADC3, ADC4, and ADC5, as well as the comparative control ADC15, are shown.

[0021] Figure 5A This is the MC38-hTF tumor growth curve, which reflects the tumor response to a single intravenous injection of 5 mg / kg of TF-targeted dual ADC 3 and the control ADC 1. The arrows indicate the administration dates.

[0022] Figure 5B This is a scatter plot of tumor volumes on day 12 after treatment with dual ADC 8 and the control ADC 15, at which point the tumors in the control group reached the study endpoint. Arrows indicate the date of administration.

[0023] Figure 6 It is the aTF ADC in Table 7 2 Time curves of the conjugate and the corresponding ADC in mouse plasma after a single intravenous (IV) administration. Invention Details This invention describes a dual-load antibody-drug conjugate comprising at least one topoisomerase inhibitor and at least one DDR inhibitor. In some embodiments, the dual-load of this invention utilizes synthetic lethality to kill cancer cells by causing DNA damage (via the topoisomerase inhibitor) and inhibiting the repair of said DNA damage (via the DDR inhibitor).

[0025] definition Unless otherwise defined, all technical terms, symbols, and other scientific terms used in this invention are intended to have the meaning commonly understood by one of ordinary skill in the art to which this invention pertains. In many cases, terms having a commonly understood meaning are defined in this invention for clarity and / or ease of reference, and such definitions included herein do not necessarily imply a difference from their commonly understood meaning in the art. The techniques and procedures described or referred to in this invention are generally well understood by one of ordinary skill in the art and are frequently employed using conventional methods, such as, for example, the widely used Green & Sambrook... Molecular Cloning: A Laboratory Manual 4 thed.(2012), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; and Ausubelet al., Current Protocols in Molecular Biology The molecular cloning methods described in John Wiley & Sons. Methods involving the use of commercially available kits and reagents are generally performed according to manufacturer-defined protocols and conditions, where appropriate, unless otherwise stated.

[0026] When referring to the compounds and conjugates provided by this invention, unless otherwise stated, the following terms have the following meanings. Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art. If there are multiple definitions for terms in this invention, unless otherwise stated, the definitions in this section shall prevail.

[0027] The singular forms “a / a / item” and “the” used in this invention include plural references unless the context clearly indicates otherwise.

[0028] The term "about / approximately" indicates and covers the value shown and the range above and below it. In some embodiments, the term "about / approximately" means ±10%, ±5%, or ±1% of the specified value. In some embodiments, the term "about / approximately" means ± one standard deviation of the specified value. In some embodiments, for example, for logarithmic scales (e.g., pH), the term "about / approximately" means ±0.3, ±0.2, or ±0.1 of the specified value.

[0029] The term "the combination thereof" includes all possible combinations of the elements referred to by the term.

[0030] The term "immunoglobulin" refers to a class of structurally related proteins that typically consist of two pairs of polypeptide chains: a pair of light (L) chains and a pair of heavy (H) chains. In a "complete immunoglobulin," all four of these chains are interconnected by disulfide bonds. The structure of immunoglobulins has been characterized in detail. See, for example, Paul, Fundamental Immunology 7th ed., Ch. 5 (2013) Lippincott Williams & Wilkins, Philadelphia, PA. In short, each heavy chain generally contains a heavy chain variable region (V... H or VH) and heavy chain constant region (C H (or CH). The heavy chain constant region generally contains three structural domains, abbreviated as C. H 1 (or CH1), C H 2 (CH2) and C H 3 (CH3). Each light chain generally contains a light chain variable region (V).L (or VL) and the light chain constant region. The light chain constant region generally contains a structural domain, abbreviated as C. L Or CL.

[0031] The term "antibody" is used in its broadest sense throughout this invention. Antibodies include complete antibodies (e.g., complete immunoglobulins) and antibody fragments (e.g., antigen-binding fragments of antibodies). An antibody contains at least one antigen-binding domain. An example of an antigen-binding domain is a V... H -V L The antigen-binding domain formed by the dimer.

[0032] V H District and V L The region can be further subdivided into high-variance regions ("HVR", also known as "complementary determinant regions (CDR)") scattered with more conservative regions. These more conservative regions are called frame regions (FR). Each V H and V L It typically consists of three CDRs and four FRs (from N-terminus to C-terminus) arranged in the following order: FR1 - CDR1 - FR2 - CDR2 - FR3 - CDR3 - FR4. CDRs participate in antigen binding and influence antigen specificity and antibody binding affinity. See Kabat et al., Sequences of Proteins of Immunological Interest 5th ed. (1991) Public Health Service, National Institutes of Health, Bethesda, MD, the entire contents of which are incorporated herein by reference.

[0033] Light chains of any vertebrate species can be classified into one of two types, called κ and λ, based on the sequence of constant structural domains.

[0034] The heavy chains of any vertebrate species can be classified into one of five distinct types (or isotypes): IgA, IgD, IgE, IgG, and IgM. These types are also named α, δ, ε, γ, and µ, respectively. IgG and IgA types are further subdivided based on sequence differences and function. Humans express the following subtypes: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.

[0035] The amino acid sequence boundaries of a CDR can be determined by any of the known numbering schemes used by someone with ordinary knowledge in the relevant technical field, including Kabat et al., ibid. (“Kabat” numbering scheme); Al-Lazikani et al., 1997. J. Mol. Biol., 273:927-948 (“Chothia” numbering scheme); MacCallum et al., 1996, J. Mol. Biol. 262:732-745 (“Contact” numbering scheme); Lefranc et al., Dev. Comp. Immunol. , 2003, 27:55-77 (“IMGT” numbering scheme); and Honeggeand Plückthun, J. Mol. Biol. Those referenced in , 2001, 309:657-70 (“AHo” numbering scheme) are determined, and each reference is incorporated herein by reference in its entirety.

[0036] Table A provides the locations of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3 identified using the Kabat and Chothia schemes. For CDR-H1, residue numbering is provided using both the Kabat and Chothia numbering schemes.

[0037] Table A. Residues in CDRs according to the Kaba and Kosia numbering scheme CDR Kabat Chothia L1 L24-L34 L24-L34 L2 L50-L56 L50-L56 L3 L89-L97 L89-L97 H1 (Kabbar ID) H31-H35B H26-H32 or H34* H1 (Coxie ID) H31-H35 H26-H32 H2 H50-H65 H52-H56 H3 H95-H102 H95-H102 * When using the Kaspersky numbering convention, the C-terminal of CDR-H1 will vary between H32 and H34 depending on the length of the CDR, such as... Figure 1 As shown.

[0038] Unless otherwise stated, the numbering scheme used to identify a specific CDR in this invention is the Kabat / Chothia numbering scheme. When the residues covered by these two numbering schemes branch (CDR-H1 and / or CDR-H2), the numbering scheme will be indicated as Kabat or Chothia. For convenience, CDR-H3 is sometimes referred to as Kabat or Chothia in this invention. However, this is not intended to imply the absence of sequence differences, and those skilled in the art can easily determine whether the sequences are the same or different by examining the sequences.

[0039] CDRs can be assigned, for example, using antibody numbering software such as Abnum, which is available from www.bioinf.org.uk / abs / abnum / and is documented in Abhinandan and Martin. Immunology The entire contents of each of the references in this paper are incorporated herein by reference in 2008, 45:3832-3839.

[0040] When referring to residues in the constant region of the antibody heavy chain, the “EU numbering scheme” is typically used (e.g., Kabat et al., as reported above). Unless otherwise stated, the EU numbering scheme is used to refer to residues in the constant region of the antibody heavy chain as described in this invention.

[0041] An "antibody fragment" contains a portion of a complete antibody, such as the antigen-binding region or variable region of the complete antibody. Antibody fragments include, for example, Fv fragments, Fab fragments, F(ab')2 fragments, Fab' fragments, scFv (sFv) fragments, and scFv-Fc fragments.

[0042] The “Fv” fragment is a non-covalently linked dimer of a heavy chain variable domain and a light chain variable domain.

[0043] In addition to the variable structural domains of the heavy and light chains, the “Fab” segment contains the constant structural domain of the light chain and the first constant structural domain (C) of the heavy chain. H1 Fab fragments can be generated, for example, by digesting full-length antibodies with papain or through recombinant methods.

[0044] The “F(ab’)2” fragment contains two Fab’ fragments linked by disulfide bonds near the hinge region. The F(ab’)2 fragment can be generated, for example, by digesting an intact antibody with pepsin or by recombinant methods. The F(ab’) fragment can be dissociated, for example, by treatment with β-mercaptoethanol.

[0045] A "single-chain Fv" or "sFv" or "scFv" antibody fragment is a V-shaped fragment contained within a single polypeptide chain. H Domain and V L Domain. V H and V L Typically, the linker is connected using a peptide linker. See Plückthun A. (1994). In some embodiments, the linker is GGGGSGGGGSGGGGS (SEQ ID NO: 75). In some embodiments, the linker is AAGSDQEPKSS (SEQ ID NO: 76). Antibodies from Escherichia coli . In Rosenberg M.&Moore GP (Eds.), The Pharmacology of Monoclonal Antibodies Vol. 113 (pp. 269-315). Springer-Verlag, New York, the entire text of which is incorporated herein by reference.

[0046] The “scFv-Fc” segment contains an scFv that is attached to the Fc structure field. For example, the Fc structure field can be attached to the C end of the scFv. The Fc structure field can be located at V.H or V L Then, depending on the directionality of the variable structural domain in scFv (i.e., V) H -V L or V L -V H Any suitable Fc domain known in the art or described herein may be used. In some cases, the Fc domain comprises the IgG1 Fc domain. In some embodiments, the IgG1 Fc domain comprises SEQ ID NO: 77 or a portion thereof. SEQ ID NO: 77 provides the C0 of the human IgG1 constant region. H 1. C H 2 and C H The sequence of 3 is: AAGSDQEPKSSDKTHTCPPCSAPELLGGSSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

[0047] The term "monoclonal antibody" refers to an antibody derived from a family of substantially homogeneous antibodies. A family of substantially homogeneous antibodies comprises antibodies that are substantially similar and bind to the same epitopes, excluding variants that normally occur during the production of monoclonal antibodies. Such variants are typically present only in small amounts. Monoclonal antibodies are generally obtained through a process involving the selection of a single antibody from a plurality of antibodies. For example, the selection process may involve choosing a unique clone from a set of multiple clones such as hybridoma clones, phage clones, yeast clones, bacterial clones, or other recombinant DNA clones. The selected antibody may be further modified, for example, to improve affinity for the target ("affinity maturation"), humanize the antibody, improve its production in cell cultures, and / or reduce its immunogenicity in subjects.

[0048] The term "chimeric antibody" refers to an antibody in which a portion of the heavy chain and / or light chain is derived from a specific source or species, while the remainder of the heavy chain and / or light chain is derived from a different source or species.

[0049] A “humanized” form of a nonhuman antibody refers to a chimeric antibody containing a minimum sequence derived from a nonhuman antibody. Humanized antibodies are typically human immunoglobulins (recipient antibodies), in which one or more CDR residues are replaced by one or more CDR residues of a nonhuman antibody (donor antibody). The donor antibody can be any suitable nonhuman antibody, such as mouse, rat, rabbit, chicken, or nonhuman primate antibodies with the desired specificity, affinity, or biological effect. In some embodiments, selected frame region residues of the recipient antibody are replaced by residues of the corresponding frame region of the donor antibody. Humanized antibodies may also contain residues not present in either the recipient or donor antibody. Such modifications can be made to further improve antibody function. See Jones et al. for further details. Nature , 1986, 321:522-525; Riechmann et al., Nature , 1988, 332: 323-329; and Presta, Curr. Op. Struct. Biol. References, 1992, 2:593-596, are incorporated herein by reference in their entirety.

[0050] "Human antibody" is an amino acid sequence having an amino acid sequence corresponding to that of an antibody produced by a human or human cell, or an antibody derived from a non-human source using a human antibody repository or a human antibody coding sequence (e.g., obtained from a human source or redesigned). Human antibodies specifically exclude humanized antibodies.

[0051] "Isolated antibody" refers to an antibody that has been separated from and / or recovered from its components in its natural environment. Components in the natural environment may include enzymes, hormones, and other proteinaceous or non-proteinaceous substances. In some embodiments, the isolated antibody is purified to a degree sufficient, for example, by using a rotor sequencer, to obtain at least 15 N-terminal or internal amino acid sequence residues. In some embodiments, the isolated antibody is purified to a degree of homogeneity as detected by gel electrophoresis (e.g., SDS-PAGE) under reducing or non-reducing conditions by Coomassie blue or silver staining. Isolated antibodies include in situ antibodies within recombinant cells, since at least one component of the antibody's natural environment is absent. In some cases, the isolated antibody is prepared by at least one purification step.

[0052] In some embodiments, the isolated antibody is purified to at least 80%, 85%, 90%, 95%, or 99% by weight. In some embodiments, the isolated antibody is purified to at least 80%, 85%, 90%, 95%, or 99% by volume. In some embodiments, the isolated antibody is provided as a solution containing at least 85%, 90%, 95%, 98%, 99%, to 100% (by volume) antibody. In some embodiments, the isolated antibody is provided as a solution containing at least 85%, 90%, 95%, 98%, 99%, to 100% (by volume) antibody.

[0053] "Affinity" refers to the sum strength of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise stated, "binding affinity" as used in this invention refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of molecule X for its partner Y can be expressed by a dissociation constant (K). D Affinity can be determined by methods commonly known in the art, including those described in this invention. Affinity can be determined using, for example, surface plasmon resonance (SPR) techniques such as Biacore. ® The instrument is used for measurement. In some implementation schemes, affinity is measured at 25°C.

[0054] Regarding antibody binding to target molecules, the terms "specific binding," "specifically binding to," "specific to," "selectively binding," and "selectively to" refer to binding to a specific antigen (e.g., a peptide target) or an epitope of a specific antigen that is assayably different from nonspecific or nonselective interactions. Specific binding can be measured, for example, by measuring the binding of a molecule against that of a control molecule. Specific binding can also be measured by competition with a control molecule that simulates the antibody binding site against the target. In this case, if the binding of the antibody to the target is competitively inhibited by the control molecule, it indicates specific binding.

[0055] "Affinity-matured" antibodies refer to antibodies with one or more alterations in one or more CDRs or FRs, said alterations resulting in increased affinity of the antibody for its antigen compared to a parent antibody without the alterations. In some embodiments, affinity-matured antibodies have nanomolar or picomolar affinity for the target antigen. Affinity-matured antibodies can be generated using a variety of methods known in the art. For example, Marks et al. ( Bio / Technology (1992, 10:779-783, the entire text of which is incorporated herein by reference) describes the invention by V H and V LAffinity maturation resulting from domain substitution. The random mutagenicity of CDRs and / or framework residues is described, for example, in Barbas et al. ( Proc. Nat. Acad. Sci. USA , 1994, 91:3809-3813); Schier et al., Gene , 1995, 169:147-155; Yelton et al., J. Immunol. , 1995, 155:1994-2004; Jackson et al., J. Immunol. , 1995, 154:3310-33199; and Hawkins et al, J. Mol. Biol. In , 1992, 226:889-896, the full text of each document is incorporated herein by reference.

[0056] The term "amino acid" refers to 20 common, naturally occurring amino acids. These include alanine (Ala; A), arginine (Arg; R), asparagine (Asn; N), aspartic acid (Asp; D), cysteine ​​(Cys; C); glutamic acid (Glu; E), glutamine (Gln; Q), glycine (Gly; G); histidine (His; H), isoleucine (Ile; I), leucine (Leu; L), lysine (Lys; K), methionine (Met; M), phenylalanine (Phe; F), proline (Pro; P), serine (Ser; S), threonine (Thr; T), tryptophan (Trp; W), tyrosine (Tyr; Y), and valine (V), as well as the less common pyrrolidone and selenocysteine. Citrulline is also included in the list of naturally occurring amino acids. Naturally encoded amino acids include 22 naturally occurring post-translational variants, such as pentadienylated amino acids, isopreneated amino acids, myristoylated amino acids, palmitoylated amino acids, etc. N -Links to glycosylated amino acids, O - Connects glycosylated amino acids, phosphorylated amino acids, and acylated amino acids. The term "amino acid" also includes "non-natural amino acids" and "modified amino acids." In this invention, the terms "non-natural amino acids" and "modified amino acids" are used interchangeably.

[0057] The term "non-natural amino acid" (or "unnatural amino acid") or "synthetic amino acid" refers to α-, β-, γ-, or δ-amino acids, and includes, but is not limited to, amino acids found in proteins, namely, glycine, alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, proline, serine, threonine, cysteine, tyrosine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, arginine, and histidine. In some embodiments, the amino acid is in the L-configuration. Alternatively, the amino acid may be a derivative of alanyl, valine, leucyl, isoleucyl, proline, phenylalanyl, tryptophanyl, methionine, glycyl, serine, threonyl, cysteine, tyrosine, asparagine, glutamine, aspartic acid, glutamate, lysine, arginine, histidine, β-alanyl, β-valine, β-leucyl, β-isoleucyl, β-proline, β-phenylalanyl, β-tryptophanyl, β-methionine, β-glycyl, β-serine, β-threonyl, β-cysteine, β-tyrosine, β-asparagine, β-glutamine, β-aspartic acid, β-glutamate, β-lysine, β-arginine, or β-histidine. Non-natural amino acids are not naturally occurring or chemically synthesized proteinogenic amino acids or their post-translational modified variants. Specifically, the term "non-natural amino acid" refers to an amino acid that is not one of the 20 common amino acids, or pyrrolidone, selenocysteine, or their post-translational modified variants. Non-limiting examples of non-natural amino acids include sulfoalanine, hydroxyproline (Hyp), β-alanine, citrulline (Cit), ornithine (Orn), leucine (Nle), 3-nitrotyrosine, nitroarginine, pyroglutamic acid (Pyr), naphthylalanine (Nal), 2,4-diaminobutyric acid (DAB), methionine sulfoxide, and methionine sulfone.

[0058] The term "modified amino acid" refers to an amino acid or non-natural amino acid containing a reactive group capable of forming a covalent bond with the linker payload. The reactive group can be amino, carboxyl, acetyl, hydrazine, acylhydrazine, hydroxylamine, aminourea, thio, azide, or alkynyl. Non-limiting examples of non-natural amino acids include: p-acetyl-L-phenylalanine, O-methyl-L-tyrosine, 3-methyl-phenylalanine, O-4-allyl-L-tyrosine, 4-propyl-L-tyrosine, fluorinated phenylalanine, isopropyl-L-phenylalanine, p-azido-L-phenylalanine, p-acyl-L-phenylalanine, p-benzoyl-L-phenylalanine, p-iodophenylalanine, p-bromophenylalanine, p-amino-L-phenylalanine, isopropyl-L-phenylalanine, p-propoxy-phenylalanine, and p-azidomethyl-L-phenylalanine.

[0059] The term "conjugate" or "antibody-drug conjugate" refers to an antibody linked to one or more payload group portions. The antibody can be any antibody described in this invention. The payload can also be any payload described in this invention. The antibody can be directly linked to the payload via a covalent bond, or the antibody can be indirectly linked to the payload via a linker. Typically, the linker is covalently linked to the antibody and also covalently linked to the payload. The term "antibody-drug conjugate" or "ADC" refers to a conjugate in which at least one payload is a therapeutic group portion, such as a drug.

[0060] The term "epitope" refers to a portion of an antigen that can specifically bind to an antibody. Epitopes often consist of surface-accessible amino acid residues and / or sugar side chains, and may possess specific three-dimensional structural features and specific charge characteristics. The difference between conformational and non-conformational epitopes is that binding to the former disappears in the presence of denaturing solvents, while binding to the latter does not. Epitopes may contain amino acid residues directly involved in binding and other amino acid residues not directly involved in binding. Epitopes bound by antibodies can be determined using known epitope assay techniques.

[0061] The percentage of "identity" between a polypeptide sequence and a reference sequence is defined as the percentage of amino acid residues in a polypeptide sequence that are identical to those in the reference sequence after alignment, and if necessary, the introduction of vacancies to achieve the maximum percentage of sequence identity. Alignment for determining the percentage of amino acid sequence identity can be achieved in various ways within the art, such as using publicly available computer software like BLAST, BLAST-2, ALIGN, MEGALIGN (DNASTAR), CLUSTALW, CLUSTAL OMEGA, or MUSCLE. Those skilled in the art can determine appropriate parameters for the alignment sequence, including any algorithm required to achieve maximum alignment of the full length of the comparison sequence.

[0062] The term "site-specific" refers to a modification of a polypeptide at a predetermined sequence position within the polypeptide. The modification is on a single, predictable residue of the polypeptide, with little or no change. In a particular embodiment, a modified amino acid is introduced at the sequence position, for example, through recombination or synthesis. Similarly, a group moiety may be "site-specifically" attached to residues at a specific sequence position in the polypeptide. In some embodiments, the polypeptide may contain more than one site-specific modification.

[0063] "Conservative substitution" or "conservative amino acid substitution" refers to the substitution of an amino acid with a chemically or functionally similar amino acid. The table of conserved substitutions provides similar amino acids well-known in the art. Peptide sequences having such substitutions are known as "conservatively modified variants." Such conservedly modified variants are complementary to polymorphic variants, interspecies homologs, and alleles, and do not exclude polymorphic variants, interspecies homologs, and alleles. For example, the amino acid groups provided in Tables B through D are considered to be conserved substitutions of each other in some embodiments.

[0064] Table B. Selected amino acid groups considered as conserved substitutions for each other in some embodiments. D and E K, R, and H S, T, N, and Q G, A, V, L and I C, M, and P F, Y and W S and T I, L, V, and M F, H, W, and Y A, C, F, G, H, I, L, M, R, T, V, W and Y D and E C, D, E, H, K, N, Q, R, S, and T H, K and R A, C, D, G, N, P, S, T and V A, G, and S A, C, D, E, G, H, K, N, Q, R, S, P and T Q, T, K, S, G, P, D, E, and R Table C. Other selected amino acid groups considered as conserved substitutions for each other in some implementation schemes. A, S, and T D and E N and Q R and K I, L and M F, Y and W Table D. Further selected groups of amino acids considered as conservative substitutions for each other in some embodiments. A and G D and E N and Q R, K and H I, L, M, V F, Y and W S and T C and M Other conservative substitutions can be found, for example, in Creighton. Proteins: Structures and Molecular Properties According to 2nd ed. (1993) WH Freeman & Co., New York, NY, antibodies produced by making one or more conserved substitutions of amino acid residues in parental antibodies are called "conservatively modified variants".

[0065] The “pAMF” mutation refers to the addition or substitution of a variant phenylalanine residue in a polypeptide, namely p-azidomethyl-L-phenylalanine.

[0066] The “pAzF” mutation refers to the addition or substitution of a variant phenylalanine residue in a polypeptide, namely p-azido-L-phenylalanine.

[0067] The "pAcF" or "pAcPhe" mutation refers to the addition or substitution of a variant phenylalanine residue, namely p-acetyl-L-phenylalanine, in a polypeptide.

[0068] The term "payload" refers to a molecular group portion that can be conjugated to an antibody. In a particular embodiment, the payload is selected from the group consisting of therapeutic group portions (e.g., topoisomerase inhibitors and DDR inhibitors described in this invention).

[0069] "Topoisomerase inhibitors" refer to any compound that interferes with topoisomerase I (Top1) or topoisomerase II (Top2), thereby inhibiting the rejoining step in the DNA unwinding process. Topoisomerase inhibitors are divided into two categories: topoisomerase I inhibitors, which include, but are not limited to, camptothecin and its derivatives (e.g., topotecan, irinotecan, rubitecan, exatecan, gly-exatecan, belotecan, and 7-ethyl-10-hydroxycamptothecin (SN-38)); and topoisomerase II inhibitors, which include, but are not limited to, etoposide, teniposide, doxorubicin, idarubicin, epirubicin, and mitoxantrone.

[0070] "DNA damage response (DDR) inhibitors" refer to any compound that can inhibit the repair of single-stranded (SSB) and / or double-stranded (DSB) DNA breaks. Cancer cells typically activate DDR pathways in their response to DNA damage, or in responses to DDR pathway defects, cancer cells may over-rely on other DDR pathways. Therefore, DDR inhibitors are widely used as anticancer drugs. DDR inhibitors include, but are not limited to, PARP1 inhibitors, ATR inhibitors, ATM inhibitors, and CHK1 / 2 inhibitors.

[0071] The term "synergistic effect" or "synergistic action" refers to the interaction of two or more drugs in which the combined effect of the two drugs is greater than the additive effect of each drug alone.

[0072] The term "linker" refers to a molecular group portion capable of forming at least two covalent bonds. Typically, a linker can form at least one covalent bond with an antibody and at least another covalent bond with a payload. In some embodiments, the linker can form more than one covalent bond with an antibody. In some embodiments, the linker can form more than one covalent bond with a payload, or multiple covalent bonds with more than one payload. After the linker forms bonds with an antibody or payload, or both, the remaining structure—that is, the residues of the linker after the formation of one or more covalent bonds—may still be referred to as the "linker" in this invention. The term "linker precursor" refers to a linker having one or more reactive groups capable of forming covalent bonds with an antibody or payload, or both. In some embodiments, the linker is a cleavable linker. For example, a cleavable linker may be a linker released through a bioinstability function, which may or may not be engineered. In some embodiments, the linker is an uncleavable linker. For example, an uncleavable linker may be a linker released after antibody degradation.

[0073] Triple-negative breast cancer (TNBC) is breast cancer characterized by estrogen receptor negativity, progesterone receptor negativity, and human epidermal growth factor receptor 2 negativity (HER2 negativity). TNBC can be BRCA1 / 2 wild-type or BRCA1 / 2 mutant. Those skilled in the art can readily determine the negative status of estrogen, progesterone, and Her2 / neu expression according to currently accepted guidelines. For example, guidelines developed by the American Society of Clinical Oncology (ASCO) and the College of American Pathologists (CAP) are widely accepted. ASCO / CAP recommends using immunohistochemistry (IHC) or in situ hybridization (ISH) techniques for detection. Furthermore, the cancer is Her2 negative if a single test (or all tests) on a tumor specimen meets any of the following criteria: (a) IHC negative, IHC 1+, or IHC 0; or (b) ISH negative using single-probe ISH or dual-probe ISH. Those skilled in the art should understand that the triple-negative cancers described in this invention do not include any cancers with significantly inconsistent histopathological features observed by a pathologist. Wolff, AC et al. J Clin Oncol. 2013 Nov. 1:31(31):3997-4013. If there is evidence that the sample expresses ER or PR and the rate of immune response in the tumor cell nucleus is <1% (see positive endogenous control), then the cancer is ER negative or PR negative.

[0074] "PARP inhibitor resistance" refers to a decrease in the efficacy of PARP inhibitors in treating, curing, or improving the condition of patients with triple-negative breast cancer. In some implementations, this PARP inhibitor resistance develops with prolonged exposure to one or more PARP inhibitors.

[0075] The "EC" used in this invention 50 "" refers to the dose, concentration, or amount of a specific test compound that elicits a dose-dependent response at 50% of the maximum expression of a specific response induced, triggered, or enhanced by the specific test compound.

[0076] The term "IC" used in this invention 50 "" refers to the amount, concentration, or dose of a specific test compound that achieves 50% inhibition of the maximum response in a test that measures this response.

[0077] The terms "subject" and "patient" used in this invention are interchangeable. The term "subject" refers to an animal, such as mammals including non-primates (e.g., cattle, pigs, horses, cats, dogs, rats, and mice) and primates (e.g., monkeys such as cynomolgus monkeys, chimpanzees, and humans), and, for example, humans. In some embodiments, the subject is refractory to or unresponsive to current treatments for hepatitis C infection. In another embodiment, the subject is a farm animal (e.g., a horse, cattle, pig, etc.) or a pet (e.g., a dog or cat). In some embodiments, the subject is a human.

[0078] As used in this invention, the terms "therapeutic agent" and "therapeutic medicine / pharmaceutical" refer to any medicine / pharmaceutical that can be used to treat or prevent a disease / condition or one or more symptoms thereof. In some embodiments, the term "therapeutic agent" includes compounds and / or antibody conjugates provided by this invention. In some embodiments, a therapeutic agent is a medicine / pharmaceutical that is known to be used, has been used, or is currently used to treat or prevent a disease / condition or one or more symptoms thereof.

[0079] As used in this invention, "therapeutic effective amount" or "effective amount" refers to the amount of an antibody or composition that, when administered to a subject, is capable of effectively treating the disease or condition. In some embodiments, the therapeutic effective amount or effective amount refers to the amount of an antibody or composition that, when administered to a subject, is capable of effectively preventing or alleviating the disease or disease progression, or alleviating symptoms. The "therapeutic effective amount" can vary depending on, in particular, the compound, the disease or condition and its severity, and the age, weight, etc., of the subject to be treated.

[0080] In some embodiments, “treatment” or “management” of any disease or condition means improving the disease or condition present in the subject. In another embodiment, “treatment” or “management” includes improving at least one physical parameter that may not be perceived by the subject. In yet another embodiment, “treatment” or “management” includes modulating the disease or condition, whether physically (e.g., stabilizing perceptible symptoms) or physiologically (e.g., stabilizing physiological parameters) or both. In another embodiment, “treatment” or “management” includes delaying or preventing the onset of the disease or condition, or delaying or preventing the recurrence of the disease or condition. In yet another embodiment, “treatment” or “management” includes reducing or eliminating the disease or condition, or delaying the progression of the disease or condition or one or more symptoms of the disease or condition, or reducing the severity of the disease or condition or one or more symptoms of the disease or condition.

[0081] The term "growth inhibition" as used in this invention (e.g., referring to cells such as tumor cells) is intended to include any measurable reduction in cell growth (e.g., tumor cell growth) upon contact with the antibody or antibody-drug conjugate described in this invention, compared to the growth of the same cells not in contact with the antibody or antibody-drug conjugate described in this invention. In some embodiments, growth may be inhibited by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99%, or 100%. The reduction in cell growth can occur through a variety of mechanisms, including but not limited to antibody internalization, apoptosis, cell necrosis, and / or effector function-mediated activity.

[0082] The terms "preventive agent" and "preventive medicine / pharmaceutical" used in this invention refer to any medicine / pharmaceutical that can be used to prevent a disease / symptom or one or more symptoms thereof. In some embodiments, the term "preventive agent" includes compounds provided by this invention. In some other embodiments, the term "preventive agent" does not refer to compounds provided by this invention. For example, a preventive agent is a medicine / pharmaceutical known to be used, or already used, or currently used to prevent or stop the onset, development, progression, and / or severity of a disease / symptom.

[0083] The phrase "preventive effective amount" as used in this invention refers to an amount of treatment / therapy (e.g., a preventive agent) sufficient to prevent or reduce the development, recurrence, or onset of one or more symptoms associated with the disease / condition, or an amount that enhances or improves the preventive effect of another treatment / therapy (e.g., another preventive agent).

[0084] The term "alkyl" as used in this invention, unless otherwise specified, refers to a saturated straight-chain or branched hydrocarbon. In some embodiments, the alkyl group is a primary, secondary, or tertiary hydrocarbon. In some embodiments, the alkyl group comprises having 1-10 carbon atoms, i.e., C1-C2. 10Alkyl. The term includes both substituted and unsubstituted group moieties, including halogenated alkyl groups. In some or any embodiments, the alkyl group is unsubstituted. In some or any embodiments, the alkyl group is substituted. In some embodiments, the alkyl group is a fluorinated alkyl group. Non-limiting examples of group moieties that can be used to replace alkyl groups are selected from the group consisting of: halogens (fluorine, chlorine, bromine, or iodine), hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate, wherein the groups are unprotected, or, if necessary, protected as known to those skilled in the art, for example in Greene, et al. The teachings of *Protective Groups in Organic Synthesis*, John Wiley and Sons, Second Edition, 1991, are incorporated herein by reference. In some embodiments, the alkyl group is selected from the group consisting of: methyl, CF3, CCl3, CFCl2, CF2Cl, ethyl, CH2CF3, CF2CF3, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl, isohexyl, 3-methylpentyl, 2,2-dimethylbutyl, and 2,3-dimethylbutyl.

[0085] As used in this invention, the term "lower alkyl" refers to a saturated straight-chain or branched hydrocarbon containing 1 to 6 carbon atoms, i.e., C1-C6 alkyl. In some embodiments, the lower alkyl group is a primary, secondary, or tertiary hydrocarbon. The term includes both substituted and unsubstituted group portions.

[0086] As used in this invention, the term "alkylene" refers to a divalent alkyl group, unless otherwise specified, as defined herein. "Substituted alkylene" refers to an alkylene group that has been substituted with respect to an alkyl group according to this invention. In some embodiments, the alkylene is unsubstituted.

[0087] As used in this invention, the term "alkenyl" refers to an olefinically unsaturated hydrocarbon group having, in some embodiments, up to about 11 carbon atoms or 2 to 6 carbon atoms (e.g., "lower alkenyl"), which may be straight-chain or branched, and has at least one or 1-2 olefinically unsaturated sites. "Substituted alkenyl" refers to an alkenyl group substituted according to the present invention with respect to an alkyl group.

[0088] As used in this invention, the term "alkenyl" refers to a divalent alkenyl group as defined herein. Lower alkenyl groups are, for example, C2-C6-alkenyl groups.

[0089] As used in this invention, the term "alkynyl" refers to an alkynyl unsaturated hydrocarbon group having, in some embodiments, up to about 11 carbon atoms or 2-6 carbon atoms (e.g., "lower alkynyl"), which may be straight-chain or branched, and has at least one or 1-2 alkynyl unsaturated sites. Non-limiting examples of alkynyl groups include alkynyl ethynyl (-C≡CH), alkynyl propyyl (-CH2C≡CH), etc. "Substituted alkynyl" refers to an alkynyl group substituted with respect to an alkyl group according to this invention.

[0090] When the term "C" x-y When used in conjunction with chemical groups such as alkyl, alkenyl, or alkynyl, it refers to a group containing x to y carbon atoms in the chain. For example, the term "C 1-6 "Alkyl" refers to a substituted or unsubstituted saturated hydrocarbon group containing 1 to 6 carbon atoms, including straight-chain alkyl and branched-chain alkyl groups. The term "C"... x-y "alkylene" refers to a substituted or unsubstituted alkylene chain containing x to y carbon atoms. For example, "-C 1-6 "alkylene-" can be selected from methylene, ethylene, propylene, butylene, pentylene, and hexylene, wherein any one of the alkylene groups may optionally be substituted. The term "C" x-y "Alkenyl" and "C" x-y "Alkyne" refers to an unsaturated aliphatic group, either substituted or unsubstituted, with a length and possible substitution similar to the aforementioned alkyl groups, but containing at least one double or triple bond. The term "-C"... x-y "-" refers to an substituted or unsubstituted alkenyl chain containing x to y carbon atoms. For example, "-C 2-6 The term "-alkenyl" can be selected from vinylene, propenene, butenene, pentenene, and hexenene, wherein any one of the alkenyl groups may be optionally substituted. The alkenyl chain may contain one or more double bonds. The term "-C" x-y "-" refers to an substituted or unsubstituted alkynyl chain containing x to y carbon atoms. For example, "-C 2-6 The '-' group can be selected from ethynylene, propynylene, butynylene, pentyynylene, and hexynylene, wherein any one of the ethynyl groups may be optionally substituted. The ethynyl chain may contain one or more triple bonds in the ethynyl chain.

[0091] The term "aryl" as used in this invention, unless otherwise specified, refers to phenyl, biphenyl, or naphthyl. The term includes both substituted and unsubstituted group portions. An aryl group may be substituted by any of the described group portions, including but not limited to one or more group portions selected from the group consisting of: halogens (fluorine (F), chlorine (Cl), bromine (Br), or iodine (I)), alkyl, haloalkyl, hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate, wherein each group is unprotected, or, if necessary, protected as known to those skilled in the art, for example in Greene, et al. , Protective Groups in Organic Synthesis As taught in John Wiley and Sons, Second Edition, 1991.

[0092] The term "aryl" as used in this invention, unless otherwise stated, refers to a divalent aryl group, wherein the aryl group is as defined in this invention.

[0093] "Alkoxy" and "alkoxy group" refer to the group –OR′, where R′ is an alkyl or cycloalkyl group. Examples of alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, etc.

[0094] "Aryloxy group" refers to the group –OR′, where R′ is an aryl group. Aryloxy groups include, for example, phenoxy groups.

[0095] "Arylalkyl" means that the alkyl group described in this invention is replaced by one or two aryl groups described in this invention.

[0096] "Alkoxycarbonyl" refers to the group -C(O)-alkoxy, wherein the alkoxy group is as defined in this invention.

[0097] "Amino" refers to the group –NH2.

[0098] As used in this invention, the term "alkylamino" unless otherwise specified refers to the group –NHR′, where R′ is C as defined in this invention. 1-10 Alkyl group. In some or any embodiment, the alkylamino group is C10. 1-6 Alkylamino.

[0099] The term "dialkylamino" as used in this invention, unless otherwise stated, refers to the group –NR′R′, wherein each R′ is independently C. 1-10 Alkyl group, as defined in this invention. In some or any embodiment, the dialkylamino group is a diC2- ... 1-6Alkylamino.

[0100] As used in this invention, the term "aminoalkyl" means, unless otherwise stated, an alkyl group as defined in this invention that is substituted with one or more amino groups. In some or any embodiments, the aminoalkyl is an alkyl group substituted with a –NH2 group (e.g., –R′(NH2), where R′ is an alkyl group as defined in this invention).

[0101] As used in this invention, "alkylaminoalkyl" refers to an alkyl group as defined in this invention, unless otherwise stated, that is substituted with one or more alkylamino groups as defined in this invention. In some embodiments, each alkyl group in alkylaminoalkyl is independently selected.

[0102] As used in this invention, "dialkylaminoalkyl" refers to an alkyl group as defined in this invention, unless otherwise stated, that is substituted with one or more dialkylamino groups as defined in this invention. In some embodiments, each alkyl group in the dialkylaminoalkyl group is independently selected.

[0103] As used in this invention, the term "hydroxyalkyl" or "hydroxyalkyl group" refers to an alkyl group as defined in this invention, unless otherwise stated. In some or any embodiments, the hydroxyalkyl group is an alkyl group substituted with a –OH group (e.g., –R′(OH), where R′ is an alkyl group as defined in this invention).

[0104] The term "haloalkyl" refers to an alkyl group as defined in this invention that is substituted with one or more halogen atoms (e.g., 1, 2, 3, 4 or 5 in some embodiments), all of which are independently selected.

[0105] "Carboxyl group" or "carboxyl group" refers to the group -C(O)OH.

[0106] The term "cycloalkyl" as used in this invention, unless otherwise specified, refers to a saturated cyclic hydrocarbon. In some embodiments, the cycloalkyl group may be a saturated, and / or bridged, and / or unbridged, and / or fused bicyclic group. In some embodiments, the cycloalkyl group comprises 3 to 10 carbon atoms, i.e., C3-C4. 10 Cycloalkyl groups. In some embodiments, the cycloalkyl group has 3 to 15 (C) groups. 3-15 ), 3 to 10 (C) 3-10 ), or 3 to 7 (C) 3-7 ) carbon atom. In some embodiments, the cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexylmethyl, cycloheptyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, decahydronaphthyl, or adamantyl.

[0107] The term "carbocyclic ring" as used in this invention, unless otherwise specified, refers to a saturated, unsaturated, or aromatic ring in which all atoms are carbon atoms. In some embodiments, the carbocyclic group may be a saturated, and / or bridged, and / or unbridged, and / or fused bicyclic group, and / or a spirocyclic bicyclic group. In some embodiments, the carbocyclic ring comprises a monocyclic ring of 3 to 10 atoms, a bicyclic ring of 6 to 12 atoms, and / or a bridged ring of 6 to 12 atoms. In some embodiments, each ring of the bicyclic carbocyclic ring may be selected from a saturated ring, an unsaturated ring, and an aromatic ring. In some embodiments, the aromatic ring (e.g., phenyl) may be fused with a saturated or unsaturated ring (e.g., cyclohexane, cyclopentane, or cyclohexene). The bicyclic carbocyclic ring includes any combination of saturated bicyclic, unsaturated bicyclic, and aromatic bicyclic rings, provided that the valence allows. Bicyclic carbocyclic rings include any combination of ring sizes, such as 4-5 fused ring systems, 5-5 fused ring systems, 5-6 fused ring systems, 6-6 fused ring systems, 5-7 fused ring systems, 6-7 fused ring systems, 5-8 fused ring systems, and 6-8 fused ring systems. In some embodiments, the carbocyclic group comprises 3-10 carbon atoms (i.e., C3-C4). 10 (Carbon ring). In some embodiments, the carbocyclic group comprises 3-12 carbon atoms (i.e., C3-C4). 12 (Carbon ring). In some embodiments, the carbon ring has 3-15 carbon atoms (C). 3-15 ), 3-12 carbon atoms (C 3-12 ), 3-10 carbon atoms (C 3-10 ), 3-7 carbon atoms (C 3-7 ), or 3-6 carbon atoms (C3-C6). In some embodiments, the carbocyclic group is cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cyclohexylmethyl, cycloheptyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, decahydronaphthyl, phenyl, indanyl, naphthyl, or adamantyl.

[0108] The terms "heterocyclic group," "heterocyclic group," and "heterocycle" refer to a monovalent monocyclic nonaromatic ring system and / or a polycyclic ring system comprising at least one nonaromatic ring, wherein one or more nonaromatic ring atoms are heteroatoms independently selected from oxygen (O), sulfur (S), and nitrogen (N); and the remaining ring atoms of the nonaromatic ring are carbon atoms, and any aromatic ring atom is an optional heteroatom independently selected from O, S, and N, while the remaining ring atoms of the nonaromatic ring are carbon atoms. In some embodiments, the heterocyclic group or heterocyclic group has 3 to 20, 3 to 15, 3 to 10, 3 to 8, 4 to 7, 4 to 11, or 5 to 6 ring atoms. The heterocyclic group can be attached to the rest of the molecule via a nonaromatic ring. In some embodiments, the heterocyclic group is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems, and wherein the N or S atom may optionally be oxidized, the N atom may optionally be quaternized, and some rings may be partially or fully saturated, or have aromaticity. The heterocyclic group may be attached to any heteroatom of the host structure or to a carbon atom of its non-aromatic ring, thereby forming a stable compound. Heterocyclic alkyl refers to a heterocyclic compound of a monovalent, monocyclic, or polycyclic non-aromatic ring system. In some or any embodiments, the heterocyclic alkyl is a monovalent, monocyclic, or polycyclic fully saturated ring system. Examples of such heterocyclic groups and / or heterocyclic alkyl groups include, but are not limited to, 2,5-diazabicyclo[2.2.2]octyl, 3,9-diazabicyclo[3.3.2]decyl, azirmonoheptanyl, benzodioxane-hexyl, benzodioxane-pentyl, benzofuranone, benzopyranone, benzopyranyl, benzotetrahydrofuranyl, benzotetrahydrothiophene, benzothiaranyl, benzooxazinyl, β-carbolinyl, chromanyl, chromone, cinnamyl, coumarinyl, decahydroisoquinolinyl, dihydrobenzisothiazinyl, dihydrobenzisothiazinyl, dihydrofuranyl, dihydroisoindolyl, dihydropyranyl, dihydropyrazolyl, dihydropyrazinyl, dihydropyridyl, dihydropyrimidin ... The heterocyclic groups include: pyrrolyl, dioxacyclopentyl, 1,4-dithiohexacyclohexyl, furanone, imidazoalkyl, imidazolinyl, indololinyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothiophenyl, isochromyl, isocoumarinyl, isoindololinyl, isothiazolyl, isoxazolyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, oxazolidinone, oxazolyl, ethylene oxide (oxiranyl), piperazinyl, piperidinyl, 4-piperidinyl, pyrazolyl, pyrazololinyl, pyrrolyl, pyrrololinyl, quininecycloyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydropyranyl, tetrahydrothiophenyl, thiomorpholinyl, thiazoalkyl, tetrahydroquinolinyl, and 1,3,5-trithiohexacyclohexyl. In some embodiments, the heterocyclic groups may also be optionally substituted as described herein.In some or any of the embodiments, the heterocyclic group and the heterocyclic alkyl group are substituted with one, two, or three groups independently selected from halogens (fluorine (F), chlorine (Cl), bromine (Br), or iodine (I)), alkyl, haloalkyl, hydroxyl, amino, alkylamino, and alkoxy groups. In some embodiments, the heterocyclic alkyl group may contain one, two, three, or four heteroatoms. Those skilled in the art will recognize that a heterocyclic alkyl group consisting of four atoms typically contains one or two heteroatoms, a heterocyclic alkyl group consisting of five or six atoms typically contains one or two heteroatoms, and a heterocyclic alkyl group consisting of seven to ten atoms typically contains one, two, three, or four heteroatoms.

[0109] The term "heteroaryl" refers to a monovalent monocyclic aromatic group and / or a polycyclic aromatic group, wherein at least one aromatic ring contains one or more heteroatoms independently selected from O, S, and N. Each ring of the heteroaryl group may contain 1 or 2 O atoms, 1 or 2 S atoms, and / or 1-4 N atoms, provided that the total number of heteroatoms in each ring is 4 or less, and each ring contains at least one carbon atom. In some embodiments, the heteroaryl group has 5-20, 5-15, or 5-10 ring atoms. The heteroaryl group may be linked to the remainder of the molecule via a nitrogen or carbon atom. In some embodiments, the monocyclic heteroaryl group includes, but is not limited to, furanyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridinyl, pyrimidinyl, pyrroleyl, imidazolyl, triazolyl, thiadiazolyl, thiazolyl, thiophene, tetrazolyl, triazinyl, and triazolyl. Examples of bicyclic heteroaryl groups include, but are not limited to, benzofuranyl, benzoimidazolyl, benzoisoxazolyl, benzopyridyl, benzothiazolyl, benzothiophene, benzotriazolyl, benzooxazolyl, furanopyridyl, imidazopyridyl, imidazothiazolyl, indoleazinyl, indole, indole, inazolyl, isobenzofuranyl, isobenzothiaphene, isoindole, isoquinolinyl, isothiazolyl, naphridyl, oxazolopyridyl, phthalazinyl, pteridinyl, purine, pyridopyridyl, pyrrolopyridyl, quinolinyl, quinoxalinyl, quinazolinyl, thiadiazopyrimidinyl, and thienenopyridyl. Examples of tricyclic heteroaryl groups include, but are not limited to, acridinel, benzoindolyl, carbazolel, dibenzofuranyl, perimidinyl, phenanthrololinyl, phenanthidyl, phenarsazinyl, phenazinyl, phenthiazinyl, phenoxazinyl, and xanthonyl. In some embodiments, the heteroaryl group may optionally be substituted as described herein. "Substituted heteroaryl" refers to a heteroaryl group substituted as defined herein with respect to aryl groups.

[0110] A "partially saturated heteroaryl" refers to a polycyclic (e.g., bicyclic, tricyclic) fused ring system comprising at least one non-aromatic ring and at least one aromatic ring, wherein one or more non-aromatic ring atoms and / or one or more aromatic ring atoms are heteroatoms independently selected from O, S, and N; and the remaining ring atoms are carbon atoms. The partially saturated heteroaryl is connected to the rest of the molecule via an aromatic ring. In some embodiments, the partially saturated heteroaryl has 6 to 20, 6 to 15, 6 to 10, 6 to 8, or 8 to 11 ring atoms. In some embodiments, the partially saturated heteroaryl group has 8, 9, 10, or 11 ring atoms (9 or 10 in some embodiments). The partially saturated heteroaryl can be attached to the host structure at any heteroatom or carbon atom of its aromatic ring to form a stable compound. In some or any embodiments, an oxo group may be present as a substituent on one of the ring atoms. Partially saturated heteroaryl groups consist of or contain one or more of the following: benzodioxane, benzodioxanepentane, benzofuranone, benzopyranone, benzopyranyl, benzotetrahydrofuranyl, benzotetrahydrothiophene, benzothiaranyl, benzooxazinyl, chromanyl, chromone, cenyl, coumarinyl, decahydroisoquinolinyl, dihydrobenzisothiazinyl, dihydrobenzisothiazinyl, dihydrofuranyl, dihydroisoindolyl, dihydropyranyl, dihydropyrazolyl, dihydropyrazinyl, tetrahydropyrazinyl, dihydropyridinone, dihydropyridyl, tetrahydropyridyl, dihydropyrimidinyl, dihydropyrroleyl, furanone, imidazolinyl. Indololinyl, tetrahydroindolyl, isoindololinyl, tetrahydroisoindolyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothiophenyl, isochromyl, isocoumarinyl, isoindololinyl, dihydroisooxazolyl, oxazinyl, dihydrooxazinyl, oxoxazolyl, dihydropiperidinoneyl, dihydro-4-piperidinoneyl, dihydropyrazolyl, dihydropyrazolinyl, dihydropyrroleyl, azabicyclo[2.2.2]oct-2-enyl, dihydrofuranyl, tetrahydroisoquinolinyl, dihydropyranyl, pyranyl, dihydrothiophenyl, oxathiazinyl, dihydrothiazolinyl, tetrahydroquinolinyl, and 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3- a ]Pyrazinyl. In some embodiments, the partially saturated heteroaryl group is benzodioxane, benzodioxanepentyl, benzofuranone, benzopyranone, benzopyranyl, benzotetrahydrofuranyl, benzotetrahydrothiophene, benzothiophene, benzooxazinyl, chromanyl, chromone, coumarinyl, dihydrobenzoisothiazinyl, dihydrobenzoisooxazinyl, dihydroisoindolyl, indololinyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothiophene, isochromanyl, isocoumarinyl, isoindololinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, or 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3- a Pyrazinyl group. In some embodiments, the partially saturated heteroaryl group may also be optionally substituted as described in this invention.

[0111] Any suitable group may be present at the “substituted” or “optionally substituted” position forming a stable molecule, including but not limited to: halogen (which may independently be F, Cl, Br, or I); cyano; hydroxyl; nitro; azide; alkoxycarbonyl; alkyl; cycloalkyl; alkenyl; alkynyl; alkoxy; aryloxy (e.g., phenoxy); alkylamino; aryl; aralkyl; or heterocyclic groups as described in this invention. In some embodiments, “optionally substituted” includes one or more substituents independently selected from halogen, alkyl, haloalkyl, hydroxyl, amino, alkylamino, and alkoxy. When the group described in this invention is optionally substituted, the optional substituents attached to the group are unsubstituted unless otherwise specified.

[0112] As used in this invention, the term "protecting group," unless otherwise stated, refers to a group added to an oxygen, nitrogen, or phosphorus atom to prevent further reaction or for other purposes. A wide variety of oxygen-protecting and nitrogen-protecting groups are known to those skilled in the art of organic synthesis.

[0113] "Pharmaceutically acceptable salt" means any salt of the compounds provided in this invention that retains the biological properties of the compounds and is non-toxic or otherwise unnecessary for medicinal use. Such salts can be derived from a variety of organic and inorganic counterions known in the art. Such salts include, but are not limited to: (1) salts reacting with organic or inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, aminosulfonic acid, acetic acid, trifluoroacetic acid, trichloroacetic acid, propionic acid, hexanoic acid, cyclopentylpropionic acid, glycolic acid, glutaric acid, pyruvic acid, lactic acid, malonic acid, succinic acid, sorbic acid, ascorbic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, picric acid, cinnamic acid, mandelic acid, phthalic acid, lauric acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphoric acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]oct-2-ene-1-carboxylic acid, glucoheponic acid, 3-Phenylacetic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfate, gluconic acid, benzoic acid, glutamic acid, hydroxynaphthylcarboxylic acid, salicylic acid, stearic acid, cyclohexylsulfamic acid, quinic acid, and mucoconic acid and similar acids form an acid addition salt; or (2) when an acidic proton is present in the parent compound, then (a) a salt formed by substitution with a metal ion such as an alkali metal ion, an alkaline earth metal ion or an aluminum ion, or an alkali metal or alkaline earth metal hydroxide such as sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, aluminum hydroxide, lithium hydroxide, zinc hydroxide and barium hydroxide, or ammonia, or (b) a salt formed with an organic base, such as an aliphatic, alicyclic or aromatic organic amine, such as ammonia, methylamine, dimethylamine, diethylamine, methylpyridine, ethanolamine, diethanolamine, triethanolamine, ethylenediamine, lysine, arginine, ornithine, choline, N, N' -Dibenzylethylenediamine, Chloroprocaine, Diethanolamine, Procaine N -Benzylphenylethylamine, N α-Methylglucosamine, piperazine, tris(hydroxymethyl)-aminomethane, tetramethylammonium hydroxide, etc., coordinate to form basic addition salts.

[0114] By way of example only and not limitation, pharmaceutically acceptable salts further include sodium, potassium, calcium, magnesium, ammonium, and tetraalkylammonium salts; when the compound contains a basic functional group, it is a salt of a non-toxic organic or inorganic acid, such as hydrohalides like hydrochlorides and hydrobroms, sulfates, phosphates, aminosulfonates, nitrates, acetates, trifluoroacetates, trichloroacetates, propionates, hexanoates, cyclopentylpropionates, glycolates, glutarates, pyruvates, lactates, malonates, succinates, sorbates, ascorbic acid salts, malates, maleates, fumarates, tartrates, citrates, benzoates, 3-(4-hydroxybenzoyl)benzoate. Salts, picrates, cinnamates, mandelates, phthalates, laurates, methanesulfonates (methanesulfonates), ethanesulfonates, 1,2-ethanedisulfonates, 2-hydroxyethanesulfonates, benzenesulfonates (benzenesulfonates), 4-chlorobenzenesulfonates, 2-naphthalenesulfonates, 4-toluenesulfonates, camphorates, camphor sulfonates, 4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylate, gluconate, 3-phenylpropionate, trimethylacetate, tert-butylacetate, lauryl sulfate, gluconate, benzoates, glutamate, hydroxynaphthylcarboxate, salicylates, stearates, cyclohexylaminosulfonates, quinates, mucoconates, etc.

[0115] With respect to the composition, the terms "substantially free" or "substantially absent" mean that the composition contains at least 85% or 90% by weight, and in some embodiments, 95%, 98%, 99%, or 100% by weight of a specified enantiomer of the compound. In some embodiments, in the methods and compounds provided by the present invention, the compound is substantially free of enantiomers.

[0116] Similarly, with respect to a composition, the term "isolated" means that the composition comprises at least 85% to 100% by weight of the compound, with the remainder comprising other chemical substances or enantiomers.

[0117] "Solvate" refers to a compound or salt thereof provided by the present invention that further comprises a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces. When the solvent is water, the solvate is a hydrate.

[0118] "Isotopic composition" refers to the amount of each isotope present for a given atom, while "natural isotopic composition" refers to the naturally occurring isotopic composition or abundance of a given atom. Atoms containing their natural isotopic composition may also be referred to as "non-enriched" atoms in this invention. Unless otherwise specified, the atoms of the compounds described in this invention are intended to represent any stable isotope of the atom. For example, unless otherwise stated, when a position is specifically designated as "H" or "hydrogen," the position is understood to be hydrogen having its natural isotopic composition.

[0119] "Isotope enrichment" refers to the percentage of a specific isotope incorporated at a given atom in a molecule, replacing the natural isotopic abundance of that atom. For example, 1% deuterium (D) enrichment at a given position means that 1% of the molecules in a given sample contain deuterium at that specified position. Since the natural distribution of deuterium is approximately 0.0156%, the deuterium enrichment at any position in a compound synthesized using non-enriched starting materials is approximately 0.0156%. The isotopic enrichment of the compounds provided by this invention can be determined using conventional analytical methods known to those skilled in the art, including mass spectrometry and nuclear magnetic resonance spectroscopy.

[0120] "Isotope-enriched" means having atoms with an isotopic composition other than the natural isotopic composition of the atoms. "Isotope-enriched" can also mean a compound containing at least one atom with an isotopic composition other than the natural isotopic composition of the atoms.

[0121] The "alkyl", "alkylene", "alkylamino", "dialkylamino", "cycloalkyl", "aryl", "alkoxy", "alkoxycarbonyl", "amino", "carboxyl", "heterocyclic", "heterocyclic", "heteroaryl", "partially saturated heteroaryl", "carboxyl" and "amino acid" groups used in this invention optionally contain deuterium (D) at one or more positions where a hydrogen atom (H) is present, and the deuterium composition of said one or more atoms is not a natural isotopic composition.

[0122] Similarly, the "alkyl", "alkylamino", "dialkylamino", "cycloalkyl", "aryl", "alkoxy", "alkoxycarbonyl", "amino", "carboxyl", "heterocyclic", "heteroaryl", "carboxyl" and "amino acid" groups used in this invention optionally contain C-13 in amounts other than the natural isotopic composition. 13 C).

[0123] In some of the chemical structures shown in this invention, certain substituents, chemical groups, and atoms are represented by curves / wavy lines (e.g., The curve / wavy line is depicted as intersecting one or more bonds to represent atoms that connect the substituents, chemical groups, and atoms. For example, in some structures, such as, but not limited to... The curve / wavy line represents the connection points where the chemical entity is connected to atoms in the main chain of the coupling structure. In some structures, for example, but not limited to... The curves / wavy lines represent the connection points where the chemical entity is connected to the atoms in the antibody or antibody fragment, and the connection points where the chemical entity is connected to the atoms in the main chain of the conjugate or linker-payload structure.

[0124] As used in this invention, the illustrations showing substituents connected to the cyclic group (e.g., aromatic, heteroaromatic, fused-ring, and saturated or unsaturated cycloalkyl or heterocycloalkyl) via bonds between ring atoms are intended to illustrate that, unless otherwise stated, the cyclic group can be substituted by the substituent at any ring position in the cyclic group or on any ring in the fused-ring group, according to the technology described in this invention or technology known in the art to which this invention pertains. For example, groups The subscripts z and y are both integers, and the bases RT and R are substituents. A The positions are general descriptions, meaning they do not directly connect to any vertex of the bond line structure, i.e., they do not directly connect to any specific ring carbon atom, including the following substituents R. A Non-limiting examples of groups that are attached to a specific ring carbon atom: .

[0125] Coupled This invention provides a dual-load antibody-drug conjugate (ADC) comprising an antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment is covalently linked via a first linker to at least one topoisomerase inhibitor and via a second linker to at least one DDR inhibitor. The conjugate comprises an antibody or antigen-binding fragment thereof that recognizes a suitable antigen (e.g., a tumor antigen), the antibody or antigen-binding fragment being covalently linked via a linker to a payload (e.g., TOPOi and DDRi as described in this invention).

[0126] The payload can be any topoisomerase inhibitor, such as Top1 inhibitors and DDR inhibitors, whichever is deemed useful by those skilled in the art. Available payloads will be described in the following sections and examples. In some embodiments, the topoisomerase inhibitor is a topoisomerase I (Top1) inhibitor. In some embodiments, the topoisomerase I inhibitor is camptothecin or a derivative thereof. In some embodiments, the topoisomerase I inhibitor is selected from the group consisting of: camptothecin, irinotecan, SN-38, topotecan, rubitecan, MLN576, belootecan, exatecan, and derivatives thereof. In one embodiment, the topoisomerase I inhibitor is selected from the group consisting of: camptothecin, irinotecan, SN-38, topotecan, exatecan, and derivatives thereof. In some embodiments, the topoisomerase I inhibitor is exatecan or a derivative thereof. In some embodiments, the topoisomerase inhibitor is a topoisomerase II inhibitor. In some embodiments, the topoisomerase II inhibitor is selected from etoposide, teniposide, tafluposide, and derivatives thereof.

[0127] In some embodiments, including any of the foregoing embodiments, the DDR inhibitor is a poly(ADP-ribose) polymerase (PARP) inhibitor, a checkpoint kinase 1 (CHK1) inhibitor, an ataxia-telangiectasia and Rad3-associated (ATR) kinase inhibitor, a nicotinamide phosphoribosyltransferase 1 (NAMPT1) inhibitor, or a RAD51 inhibitor. In some embodiments, including any of the foregoing embodiments, the DDR inhibitor is a PARP inhibitor. In some embodiments, including any of the foregoing embodiments, the PARP inhibitor is selected from olaparib, veliparib, talazoparib, rucaparib, niraparib, AZD5305, talzenna, and derivatives thereof. In some embodiments, including any of the foregoing embodiments, the PARP inhibitor is talazoparib or a derivative thereof.

[0128] Both the first and second linkers can be any linker capable of forming at least one bond with the antibody and at least one bond with the payload, wherein the first linker forms at least one bond with either the topoisomerase inhibitor or the DDR inhibitor, and the second linker forms a bond with the other of the topoisomerase inhibitor or the DDR inhibitor. The first and second linkers may have the same structure. In some embodiments, the first and second linkers have different structures. In some embodiments, including any of the foregoing embodiments, the first and second linkers each independently comprise a protease-cleavable linker, an enzyme-cleavable linker, a pH-sensitive linker, or an uncleavable linker. In some embodiments, the first and second linkers each independently comprise a β-glucuronide cleavable by β-glucuronidase. In some embodiments, the first and second linkers each independently comprise a cathepsin-cleavable linker.

[0129] Available linkers include those described in this invention. In some embodiments, the first and second linkers are each independently any divalent or multivalent linker known to those skilled in the art. Available divalent linkers include alkylene, substituted alkylene, heteroalkylene, substituted heteroalkylene, arylene, substituted arylene, heteroarylene, and substituted heteroarylene. In some embodiments, the first linker and / or the second linker are C10 and C20, respectively. 1-10 Alkylene or C 1-10 Heteroalkylene. In some embodiments, the C 1-10 Heteroalkylene compounds are polyethylene glycol (PEG).

[0130] In some embodiments, both the first and / or second linkers are hydrolyzed stable. A hydrolyzed stable linker is one that is substantially stable in water and does not react with water at a useful pH (including, but not limited to, physiological conditions), and this stability can persist for a considerable period, possibly indefinitely. In some embodiments, both the first and / or second linkers are hydrolyzed unstable. A hydrolyzed unstable or degradable linker is one that is degradable in water or aqueous solutions (including, for example, blood). An enzymatically unstable or degradable linker is one that can be degraded by one or more enzymes.

[0131] As understood in the art, polyethylene glycol (PEG) and related polymers may contain degradable linkers in the polymer backbone or in linker groups between the polymer backbone and one or more terminal functional groups of the polymer molecule. For example, ester bonds formed by the reaction of PEG carboxylic acid or activated PEG carboxylic acid with alcohol groups on a bioactive agent typically hydrolyze under physiological conditions, thereby releasing the drug.

[0132] Other hydrolyzable bonds include, but are not limited to, carbonate bonds; imine bonds formed by the reaction of amines and aldehydes; phosphate ester bonds formed by the reaction of alcohols and phosphate groups; hydrazone bonds formed by the reaction of hydrazides and aldehydes; acetal bonds formed by the reaction of aldehydes and alcohols; orthoester bonds formed by the reaction of formate esters and alcohols; peptide bonds formed by amine groups (including, but not limited to, amine groups at the end of polymers (e.g., PEG)) and carboxyl groups of peptides; and oligonucleotide bonds formed by phosphoramidite groups (including, but not limited to, phosphoramidite groups at the end of polymers) and 5' hydroxyl groups of oligonucleotides.

[0133] Various cleavable linkers are known to those skilled in the art. See U.S. Patent Nos. 4,618,492, 4,542,225, and 4,625,014. Mechanisms for releasing drugs from these linker groups include, for example, irradiation of the photosensitive bond and acid-catalyzed hydrolysis. For example, U.S. Patent No. 4,671,958 describes an immunoconjugate comprising linkers that can be cleaved by proteolytic enzymes of the patient's complement system at a target site in vivo. The length of the linker can be predetermined or selected based on the desired spatial relationship between the peptide and the molecule to which it is linked. Given the numerous methods reported for linking various radiodiagnostic compounds, radiotherapeutic compounds, drugs, toxins, and other agents to peptides, those skilled in the art will be able to determine suitable methods for linking a given drug to a peptide.

[0134] The linkers described in this invention can have a wide range of molecular weights or molecular lengths. Linkers with larger or smaller molecular weights can be used to achieve the desired spatial relationship or conformation between the peptide and the linked entity. Linkers with longer or shorter molecular lengths can also be used to achieve the desired spatial or flexible relationship between the peptide and the linked entity. Similarly, linkers with specific shapes or conformations can be used to impart a specific shape or conformation to the peptide or the linked entity before or after it reaches its target site. The functional groups at each end of the linker can be selected as needed to regulate the release of the peptide or payload under desired conditions. This optimization of the spatial relationship between the peptide and the linked entity can endow the molecule with new, tunable, or desired properties.

[0135] In some embodiments, the present invention provides a water-soluble bifunctional linker having a dumbbell-shaped structure, comprising: a) a portion of an azide, alkyne, hydrazine, acylhydrazine, hydroxylamine, or carbonyl-containing group attached to at least a first end of a polymer backbone; and b) a portion of at least a second functional group attached to a second end of the polymer backbone. The second functional group may be the same as or different from the first functional group. In some embodiments, the second functional group does not react with the first functional group. In some embodiments, the present invention also provides a water-soluble compound comprising at least one branched molecular structure arm. For example, the branched molecular structure may be a dendritic structure.

[0136] In some implementations, the first and / or second connectors are each independently derived from a connector precursor, which is selected from the group consisting of: N 3-Succinimide-3-(2-pyridinedithio)propionate (SPDP) N succinimide-4-(2-pyridinedithio)valerate (SPP) N succinimide-4-(2-pyridinedithio)butyrate (SPDB) N -Succinimide-4-(2-pyridinedithio)-2-sulfonylbutyrate (sulfon-SPDB) N - Succinimide iodoacetate (SIA) N succinimide (4-iodoacetyl)aminobenzoate (SIAB), maleimide PEG NHS, N 4-(maleimide methyl)cyclohexane carboxylate (SMCC) N -Sulfosuccinimide-4-(maleimide-methyl)cyclohexanecarboxylate (sulfon-SMCC), or 2,5-dioxopyrrolidone-1-yl17-(2,5-dioxo-2,5-dihydro-1-yl) H (-pyrrolo-1-yl)-5,8,11,14-tetraoxo-4,7,10,13-tetraazaheptadecane-1-ester (CX1-1). In a particular embodiment, the linker is derived from a linker precursor. N -Succinimide-4-(maleimide-methyl)cyclohexanecarboxylate (SMCC).

[0137] In some embodiments, the first and / or second linkers are each derived from a linker precursor selected from the group consisting of dipeptides, tripeptides, tetrapeptides, and pentapeptides. In such embodiments, the linkers are cleavable by a protease. Exemplary dipeptides include, but are not limited to, valine-citrulline (VC or val-cit), alanine-phenylalanine (AF or ALA-Phe), phenylalanine-lysine (FK or Phe-lys), phenylalanine-homolysine (Phe-homolys), and... N -Methyl-valine-citrulline (Me-val-cit). Exemplary tripeptides include, but are not limited to, gly-valine-citrulline (gly-val-cit), gly-gly-gly (gly-gly-gly), and gly-(methoxyethoxyethyl)serine-valine (gly-val-citalanine OMESerValAla).

[0138] In some embodiments, the first and / or second linkers of the present invention each comprise a self-immolative spacer group. In some embodiments, the self-immolative spacer group comprises a p-aminobenzyl group. In some embodiments, p-aminobenzyl alcohol is linked to the amino acid unit via an amide bond, and a carbamate, methyl carbamate, or carbonate (Hamann's ester) is formed between the benzyl alcohol and the payload. et al (2005) Expert Opinion. Ther. Patents (2005) 15:1087-1103. In some embodiments, the linker comprises a p-aminobenzyloxycarbonyl (PAB). Other examples of self-cleaving spacer groups include, but are not limited to, aromatic compounds with electronic properties similar to the PAB group, such as 2-aminoimidazolium-5-methanol derivatives (US Patent No. US7,375,078; Hay). et al (1999) Bioorg. Med. Chem. Lett. 9:2237) and o-aminobenzyl acetal or p-aminobenzyl acetal. In some embodiments, spacer groups that undergo cyclization upon hydrolysis of the amide bond may be used, such as substituted and unsubstituted 4-aminobutyric acid amides (Rodrigues). et al. (1995) Chemistry Biology 2:223), appropriately substituted bicyclic [2.2.1] and bicyclic [2.2.2] ring systems (Storm) et al. (1972) J. Amer. Chem. Soc. 94:5815) and 2-aminophenylpropionic acid amides (Amsberry, et al.(1990) J. Org. Chem. 55:5867). Linking a drug to the α-carbon of a glycine residue is another example of a self-cleaving spacer group, which may be useful for conjugates (Kingsbury). et al. (1984) J. Med. Chem. 27:1447).

[0139] In some embodiments, the linker precursors can be combined to form larger linkers. For example, in some embodiments, the linker comprises a dipeptide valine-citrulline and a p-aminobenzyloxycarbonyl group. These linkers are also referred to as citValCit-PAB linkers.

[0140] Antibodies are typically proteins comprising multiple polypeptide chains. In some embodiments, antibodies are heterotetramers comprising two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain via a covalent disulfide bond. Each heavy chain is linked to another heavy chain via one or more covalent disulfide bonds. Each heavy chain and each light chain may also have one or more intrachain disulfide bonds. As known to those skilled in the art, each heavy chain typically contains a variable domain (V... H Then there are multiple constant structural domains. Each light chain typically contains a variable structural domain (V0) at one end. L ), and contains constant structural domains. As is known to those skilled in the art, antibodies typically have selective affinity for their target molecules, namely antigens.

[0141] The antibodies provided by this invention can be in any antibody form known to those skilled in the art. They can be full-length or fragments. Exemplary full-length antibodies include IgA, IgA1, IgA2, IgD, IgE, IgG, IgG1, IgG2, IgG3, IgG4, IgM, etc. Exemplary fragments include Fv, Fab, Fc, scFv, scFv-Fc, etc.

[0142] In some embodiments, the antibody of the conjugate comprises 1, 2, 3, 4, 5, or 6 CDR sequences as described in this invention. In some embodiments, the antibody of the conjugate comprises the heavy chain variable domain (V) as described in this invention. H In some embodiments, the antibody of the conjugate comprises the light chain variable domain (V) described in this invention. L In some embodiments, the antibody of the conjugate comprises the heavy chain variable domain (V) described in this invention. H ) and the light chain variable structural domain (V) described in this invention L In some embodiments, the antibody of the conjugate comprises the paired heavy chain variable domain and light chain variable domain (V) as described in this invention. H - VL (Yes). In some embodiments, the antibody of the conjugate comprises three heavy chain CDRs and three light chain CDRs, the sequence of which consists of heavy chain and light chain CDRs of any paired heavy chain variable region and light chain variable region as described in this invention (any V). H -V L (The six CDRs are correct).

[0143] In some embodiments, the antibody of the conjugate comprises any amino acid sequence of the antibody of the present invention. In some embodiments, the antibody comprises any of the above-described amino acid sequences having up to 10 amino acid substitutions. In some embodiments, the antibody comprises any of the above-described amino acid sequences having up to 9 amino acid substitutions. In some embodiments, the antibody comprises any of the above-described amino acid sequences having up to 8 amino acid substitutions. In some embodiments, the antibody comprises any of the above-described amino acid sequences having up to 7 amino acid substitutions. In some embodiments, the antibody comprises any of the above-described amino acid sequences having up to 6 amino acid substitutions. In some embodiments, the antibody comprises any of the above-described amino acid sequences having up to 5 amino acid substitutions. In some embodiments, the antibody comprises any of the above-described amino acid sequences having up to 4 amino acid substitutions. In some embodiments, the antibody comprises any of the above-described amino acid sequences having up to 3 amino acid substitutions. In some embodiments, the antibody comprises any of the above-described amino acid sequences having up to 2 amino acid substitutions. In some embodiments, the antibody comprises any of the above-described amino acid sequences having up to 1 conserved amino acid substitution. In some embodiments, all amino acid substitutions are conserved amino acid substitutions. For example, in some embodiments, the antibody comprises any of the above-described amino acid sequences having up to 10 conserved amino acid substitutions. In some embodiments, the antibody comprises any of the above-mentioned amino acid sequences having up to 9 conserved amino acid substitutions. In some embodiments, the antibody comprises any of the above-mentioned amino acid sequences having up to 8 conserved amino acid substitutions. In some embodiments, the antibody comprises any of the above-mentioned amino acid sequences having up to 7 conserved amino acid substitutions. In some embodiments, the antibody comprises any of the above-mentioned amino acid sequences having up to 6 conserved amino acid substitutions. In some embodiments, the antibody comprises any of the above-mentioned amino acid sequences having up to 5 conserved amino acid substitutions. In some embodiments, the antibody comprises any of the above-mentioned amino acid sequences having up to 4 conserved amino acid substitutions. In some embodiments, the antibody comprises any of the above-mentioned amino acid sequences having up to 3 conserved amino acid substitutions. In some embodiments, the antibody comprises any of the above-mentioned amino acid sequences having up to 2 conserved amino acid substitutions. In some embodiments, the antibody comprises any of the above-mentioned amino acid sequences having up to 1 conserved amino acid substitution.

[0144] In some embodiments, the antibody conjugate may be formed from an antibody comprising one or more reactive groups. In some embodiments, the antibody conjugate may be formed from an antibody comprising all naturally encoded amino acids. Those skilled in the art will recognize that several naturally encoded amino acids include active groups capable of being conjugated to a payload or a linker. These active groups include cysteine ​​side chains, lysine side chains, and amino-terminal groups suitable for conjugation. In these embodiments, the antibody conjugate may comprise a payload or linker linked to residues of an antibody active group. In these embodiments, the payload precursor or linker precursor comprises an active group capable of forming a bond with the antibody active group. Typical active groups include maleimide groups, activated carbonates (including but not limited to p-nitrobenzene esters), and activated esters (including but not limited to...). N -Hydroxysuccinimide, p-nitrobenzene ester, and aldehydes). Particularly useful active groups include maleimide and succinimide, for example... N -Hydroxysuccinimide is used to form bonds with cysteine ​​and lysine side chains. Other active groups include alkynes (e.g., strained alkynes), azides, and aminooxy groups, used to form bonds with non-natural amino acids incorporated into the antibody polypeptide chain. Further active groups are described in the following sections and examples.

[0145] In some embodiments, the antibody comprises one or more modified amino acids or non-natural amino acids having an active group, as described in this invention. Typically, the modified amino acids or non-natural amino acids are not naturally encoded amino acids. These modified amino acids may contain an active group for forming a covalent bond with a linker precursor or a payload precursor. Those skilled in the art can use the active group to link peptides to any molecular entity capable of forming a covalent bond with the modified amino acid. Therefore, this invention provides conjugates comprising antibodies containing modified amino acid residues directly or indirectly linked to a payload via a linker. Exemplary modified amino acids are described in the following sections. Typically, the modified amino acid has an active group capable of forming a bond with a linker or payload having a complementary active group.

[0146] Non-natural or modified amino acids are located at selected positions in the antibody's polypeptide chain. These positions are identified as optimal sites for providing non-natural or modified amino acid substitutions. Each site is capable of carrying a non-natural or modified amino acid that provides optimal structure, function, and / or method of antibody generation.

[0147] In some embodiments, the site-specific location used for substitution provides a stable antibody. Stability can be determined by any technique readily apparent to those skilled in the art.

[0148] In some embodiments, the site-specific position used for substitution provides an antibody with optimal functional properties. For example, the antibody exhibits little or no loss of binding affinity to its target antigen compared to antibodies lacking site-specific non-natural amino acids or modified amino acids. In some embodiments, the antibody may show enhanced binding compared to antibodies lacking site-specific non-natural amino acids or modified amino acids.

[0149] In some embodiments, the site-specific position used for substitution provides an antibody that can be advantageously produced. For example, in some embodiments, the antibody exhibits advantageous properties in its synthesis method, as described below. In some embodiments, the antibody shows little or no loss in production yield compared to antibodies without site-specific non-natural amino acids. In some embodiments, the antibody may show increased production yield compared to antibodies without site-specific non-natural amino acids or modified amino acids. In some embodiments, the antibody may show little or no loss of tRNA inhibition compared to antibodies without site-specific non-natural amino acids or modified amino acids. In some embodiments, the antibody may show enhanced tRNA inhibition during production compared to antibodies without site-specific non-natural amino acids or modified amino acids.

[0150] In some embodiments, the site-specific position for substitution provides an antibody with favorable solubility. In some embodiments, the antibody exhibits little or no loss of solubility compared to antibodies containing site-specific non-natural amino acids or modified amino acids. In some embodiments, the antibody may show enhanced solubility compared to antibodies containing site-specific non-natural amino acids or modified amino acids.

[0151] In some embodiments, the site-specific position used for substitution provides an antibody with favorable expression. In some embodiments, the antibody may show little or no loss of expression compared to antibodies that do not contain site-specific non-natural amino acids or modified amino acids. In some embodiments, the antibody may show enhanced expression compared to antibodies that do not contain site-specific non-natural amino acids or modified amino acids.

[0152] In some embodiments, the site-specific position for substitution provides an antibody with favorable folding properties. In some embodiments, the antibody may exhibit little or no loss in proper folding compared to antibodies without site-specific non-natural amino acids or modified amino acids. In some embodiments, the antibody may exhibit enhanced folding properties compared to antibodies without site-specific non-natural amino acids or modified amino acids.

[0153] In some embodiments, the site-specific position for substitution provides an antibody capable of favorable conjugation. As described below, several non-natural or modified amino acids have side chains or functional groups that can directly or indirectly facilitate antibody conjugation to a second drug via a linker. In some embodiments, the antibody may exhibit enhanced conjugation efficiency compared to antibodies that do not have the same or other non-natural or modified amino acids at other positions. In some embodiments, the antibody may exhibit enhanced conjugation yield compared to antibodies that do not have the same or other non-natural or modified amino acids at other positions. In some embodiments, the antibody may exhibit enhanced conjugation specificity compared to antibodies that do not have the same or other non-natural or modified amino acids at other positions.

[0154] One or more non-natural or modified amino acids are located at selected site-specific positions in at least one polypeptide chain of the antibody. The polypeptide chain can be any polypeptide chain of the antibody, without limitation, including light or heavy chains. The site-specific positions can be located in any domain of the antibody, including any variable domain and any constant domain.

[0155] In some embodiments, the antibody provided by the present invention contains one non-natural or modified amino acid at a site-specific position. In some embodiments, the antibody provided by the present invention contains two non-natural or modified amino acids at a site-specific position. In some embodiments, the antibody provided by the present invention contains three non-natural or modified amino acids at a site-specific position. In some embodiments, the antibody provided by the present invention contains three or more non-natural or modified amino acids at a site-specific position. In some embodiments, the antibody provided by the present invention contains four non-natural or modified amino acids at a site-specific position.

[0156] In some embodiments, the antibody provided by the present invention comprises one or more non-natural amino acids or modified amino acids, each located at a position selected from the group consisting of the following heavy chain residues or light chain residues: HC-F404, HC-K121, HC-Y180, HC-F241, HC-221, HC-Y391, LC-T22, LC-S7, LC-N152, LC-K42, LC-E161, LC-D170, HC-S136, HC-S25, HC-A40, HC-S119, HC-S190, HC-K222, HC-R19, HC-Y52, or HC-S70, or their translated modified variants, according to the Kabat, Chothia, or EU numbering schemes. In some embodiments, the antibody provided by the present invention comprises one or more non-natural amino acids or modified amino acids, each located at a position selected from the group consisting of the following heavy chain residues or light chain residues: HC-F404, HC-Y180, HC-F241, HC-Y391, LC-K42, and LC-E161, or their translated modified variants, according to the Kabat, Chothia, or EU numbering schemes. In some embodiments, the antibody provided by the present invention comprises one or more non-natural amino acids or modified amino acids, each located at a position selected from the group consisting of the following heavy chain residues or light chain residues: HC-F404, HC-F241, and LC-K42, or their translated modified variants, according to the Kabat, Chothia, or EU numbering schemes. In some embodiments, the antibody provided by the present invention comprises one or more non-natural or modified amino acids, each located at a position selected from the group consisting of the following heavy chain residues or light chain residues: HC-F404 and HC-F241 according to the Kabat, Chothia, or EU numbering schemes, or their translated modified variants. In some embodiments, the antibody provided by the present invention comprises one or more non-natural or modified amino acids, each located at a position selected from the group consisting of the following heavy chain residues or light chain residues: HC-F404 and LC-K42 according to the Kabat, Chothia, or EU numbering schemes, or their translated modified variants.In some embodiments, the antibody provided by the present invention comprises one or more non-natural amino acids or modified amino acids, each located at a position selected from the group consisting of heavy chain residues or light chain residues: HC-F241 and LC-K42 according to the Kabat, Chothia, or EU numbering schemes, or post-translational modified variants thereof. In some embodiments, the antibody provided by the present invention comprises a non-natural amino acid or modified amino acid at position HC-F404 according to the Kabat, Chothia, or EU numbering schemes, or a post-translational modified variant thereof. In some embodiments, the antibody provided by the present invention comprises a non-natural amino acid or modified amino acid at position HC-F241 according to the Kabat, Chothia, or EU numbering schemes, or a post-translational modified variant thereof. In some embodiments, the antibody provided by the present invention comprises a non-natural amino acid or modified amino acid at position HC-Y180 according to the Kabat, Chothia, or EU numbering schemes, or a post-translational modified variant thereof. In some embodiments, the antibody provided by the present invention comprises a non-natural amino acid or a modified amino acid, or a post-translational variant thereof, at positions HC-F404 and HC-Y180 according to the Kabat, Chothia, or EU numbering scheme. In some embodiments, the antibody provided by the present invention comprises a non-natural amino acid or a modified amino acid, or a post-translational variant thereof, at position HC-F241 according to the Kabat, Chothia, or EU numbering scheme. In some embodiments, the antibody provided by the present invention comprises a non-natural amino acid or a modified amino acid, or a post-translational variant thereof, at position HC-Y391 according to the Kabat, Chothia, or EU numbering scheme. In some embodiments, the antibody provided by the present invention comprises a non-natural amino acid or a modified amino acid, or a post-translational variant thereof, at position LC-K42 according to the Kabat, Chothia, or EU numbering scheme. In some embodiments, the antibody provided by the present invention comprises a non-natural amino acid or a modified amino acid, or a post-translational variant thereof, at position LC-E161 according to the Kabat, Chothia, or EU numbering scheme. In some embodiments, the antibody provided by the present invention comprises a non-natural amino acid or a modified amino acid, or a post-translational variant thereof, at positions HC-F404, HC-Y180, and LC-K42 according to the Kabat, Chothia, or EU numbering scheme.In some embodiments, the antibody provided by the present invention comprises non-natural or modified amino acids, or post-translational variants thereof, at positions HC-F404, HC-Y180, LC-K42, and LC-E161 according to the Kabat, Chothia, or EU numbering scheme. In some embodiments, the antibody provided by the present invention comprises non-natural or modified amino acids, or post-translational variants thereof, at positions HC-F404, HC-Y180, and HC-F241 according to the Kabat, Chothia, or EU numbering scheme. In some embodiments, the antibody provided by the present invention comprises non-natural or modified amino acids, or post-translational variants thereof, at positions HC-F404, HC-Y180, HC-F241, and LC-K42 according to the Kabat, Chothia, or EU numbering scheme. In some embodiments, the antibodies provided by this invention comprise non-natural or modified amino acids, or post-translational variants thereof, at positions HC-F404, HC-Y180, HC-F241, and LC-K42 according to the Kabat, Chothia, or EU numbering schemes. In these designations, HC represents heavy chain residues, and LC represents light chain residues.

[0157] In some embodiments, the antibody conjugate has the structure shown in Formula I: Formula (I) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof; Where Ab is the antibody or its antigen-binding fragment; L a It is the first connector; L b It is the second connector; Each DDRi is an independent residue of a DDR inhibitor; Each TOPOi is an independent residue of a topoisomerase inhibitor; The subscript n is an integer selected from 1 to 10; and The subscript m is an integer selected from 1 to 10.

[0158] In some embodiments of Formula (I), the topoisomerase inhibitor TOPOi is a topoisomerase I (Top1) inhibitor. In some embodiments of Formula (I), the topoisomerase I inhibitor is camptothecin or a derivative thereof. In some embodiments of Formula (I), the topoisomerase I inhibitor is selected from the group consisting of: camptothecin, irinotecan, SN-38, topotecan, rubitecan, MLN576, belotetcan, ixenonotecan, and derivatives thereof. In some embodiments of Formula (I), the topoisomerase I inhibitor is selected from the group consisting of: camptothecin, irinotecan, SN-38, topotecan, ixenonotecan, and derivatives thereof. In some embodiments of Formula (I), the topoisomerase I inhibitor is ixenonotecan or a derivative thereof. In some embodiments of Formula (I), the topoisomerase inhibitor is a topoisomerase II inhibitor. In some embodiments of Formula (I), the topoisomerase II inhibitor is selected from etoposide, teniposide, tafluposide, and derivatives thereof.

[0159] In some embodiments of Formula (I), including any of the foregoing embodiments, the DDR inhibitor DDRi is a poly(ADP-ribose) polymerase (PARP) inhibitor, a checkpoint kinase 1 (CHK1) inhibitor, an ataxia-telangiectasia and Rad3-associated (ATR) kinase inhibitor, a nicotinamide phosphoribosyltransferase 1 (NAMPT1) inhibitor, or a RAD51 inhibitor, or a derivative thereof. In some embodiments of Formula (I), including any of the foregoing embodiments, the DDR inhibitor is a PARP inhibitor. In some embodiments of Formula (I), including any of the foregoing embodiments, the PARP inhibitor is selected from olaparib, veliparib, talazoparib, rucaparib, niraparib, AZD5305, talzenna, and derivatives thereof. In some embodiments of Formula (I), including any of the foregoing embodiments, the PARP inhibitor is talazoparib or a derivative thereof.

[0160] In some embodiments of formula (I), including any of the foregoing embodiments, the connector L a and L b Each linker independently comprises a protease-cleavable linker, an enzyme-cleavable linker, a pH-sensitive linker, or a non-cleavable linker. In some embodiments, the first and second linkers each independently comprise a β-glucuronide cleavable by β-glucuronidase. In some embodiments, the first and second linkers each independently comprise a cathepsin-cleavable linker.

[0161] In some embodiments, the antibody conjugate represented by formula (I) is as shown in formula (Ia-1): Equation (Ia-1) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof; wherein: Ab is the antibody or its antigen-binding fragment; Each DDRi is an independent residue of a DDR inhibitor; Each TOPOi is an independent residue of a topoisomerase inhibitor; Each SP 1 SP 2 and SP 3 Each time it appears, it is either independently absent or independently a divalent spacer group; Each W 1 Each time it appears, it either does not exist independently, or it is independently... Wherein -NH- is connected to W 2 ; and each These respectively represent the connection points that connect to the rest of the general formula; L 1 Independently as a key, or L 1 C can be substituted independently. 1-6 alkylene, wherein the C 1-6 The alkylene group is optionally substituted by one, two or three substituents selected from halogen, alkyl, haloalkyl, hydroxyl, amino, alkylamino and alkoxy groups; Each W 2 Each time it appears, it is either independently absent or independently an amino acid residue or a peptide residue, wherein the amino acid residue or the peptide residue is optionally represented by HP. 2 Group substitution; Each HP 1 Each time it appears, it is either independently absent or independently a divalent hydrophilic group; Each HP 2 When present, they are monovalent hydrophilic groups; Each R A Each time it appears, it is independently represented by an arbitrarily substituted C. 1-6 Alkyl, wherein the C 1-6 The alkyl group is optionally substituted by one, two or three substituents selected from halogens, alkyl groups, haloalkyl groups, hydroxyl groups, amino groups, alkylamino groups and alkoxy groups; Each RT is either independently absent or independently a release trigger group each time it appears. Each RL is a reactive linker. y is an integer independently selected from 0, 1, and 2; and z is an integer independently selected from 0 and 1.

[0162] In some embodiments, the antibody conjugate represented by formula (Ia-1) is as shown in formula (Ia): Formula (Ia) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof; wherein: Ab is the antibody or its antigen-binding fragment; Each DDRi is an independent residue of a DDR inhibitor; Each TOPOi is an independent residue of a topoisomerase inhibitor; Each SP 1 SP 2 and SP 3 Each time it appears, it is either independently absent or independently a divalent spacer group; Each W 1 Each time it appears, it either does not exist independently, or it is independently... Wherein -NH- is connected to W 2 ; and each These represent the connection points that connect to the rest of the general formula.

[0163] L 1 Independently as a key, or L 1 C can be substituted independently. 1-6 alkylene, wherein the C 1-6 The alkylene group is optionally substituted by one, two or three substituents selected from halogen, alkyl, haloalkyl, hydroxyl, amino, alkylamino and alkoxy groups; Each W 2 Each time it appears, it is either independently absent or independently an amino acid residue or a peptide residue, wherein the amino acid residue or the peptide residue is optionally represented by HP. 2 Group substitution; Each HP 1 Each time it appears, it is either independently absent or independently a divalent hydrophilic group; Each HP 2 When present, they are monovalent hydrophilic groups; Each R A Each time it appears, it is independently represented by an arbitrarily substituted C. 1-6Alkyl, wherein the C 1-6 The alkyl group is optionally substituted by one, two or three substituents selected from halogens, alkyl groups, haloalkyl groups, hydroxyl groups, amino groups, alkylamino groups and alkoxy groups; Each RT is either independently absent or independently a release trigger group each time it appears. Each RL is a reactive linker. y is an integer independently selected from 0, 1, and 2; and z is an integer independently selected from 0 and 1.

[0164] In some embodiments, the antibody conjugate shown in formula (I) is as shown in formula (Ib): Formula (Ib) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof; wherein: L 2 It is -C 1-6 alkylene-, the -C 1-6 The alkylene group is optionally substituted with one or more substituents selected from the following: halogen, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; Y is –X 1 -C 1-6 Alkylene-[X] 1 -C 1-6 [alkylene] a -[X 1 ] b –、–X 1 -C 2-6 imide-[X] 1 -C 2-6 [Alkenyl] a -[X 1 ] b – or –X 1 -C 2-6 Ethyne-[X] 1 -C 2-6 [Iso-ynyl]a -[X 1 ] b – wherein at least one alkylene, alkenylene, or ynylene group in Y is selected from one or more groups chosen from R. 50 Substituents and The alkylene, alkenylene, or ynylene group in Y is optionally selected from one or more groups selected from R. 51 Substituents of the substituents; R 50 Yes –C 1-6 Alkylene-X 2 -[C 1-6 [alkylene] c -HP 2 –C 2-6 imide-X 2 -[C 2-6 [Alkenyl] c -HP 2 、or –C 2-6 Ethyne-X 2 -[C 2-6 [Iso-ynyl] c -HP 2 , where R 50 Each alkylene, alkenylene, or ynylene group may optionally be substituted by one or more substituents selected from the following: halogen, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; R 51 Independently selected from halogens, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; X 1 and X 2 Each is independently selected from –N(R) 10–, –C(O)–, and –N(R)– 10 )C(O)–; R 10 Each time it appears, it is independently selected from H and C. 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; a is an integer selected from 0, 1, 2, and 3; b is an integer selected from 0 and 1; c is an integer selected from 0 and 1; Su is the hexose form of the monosaccharide; and DDRi, SP 1 W 1 W 2 HP 1 HP 2 SP 3 , RL, Ab and TOPOi all have the definition described in equation (Ia).

[0165] In some embodiments, the antibody conjugate represented by formula (Ib) is as shown in formula (Ib-A): Formula (Ib-A) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

[0166] In some embodiments, the antibody conjugate represented by formula (Ib) is as shown in formula (Ib-B): Formula (Ib-B) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

[0167] In some embodiments, the antibody conjugate shown in formula (I) is as shown in formula (Ic): Formula (Ic) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof; wherein: DDRi, SP 1 W 1 W 2 HP 1 HP 2 SP 3RL, Y, L 2 Su, Ab, TOPOi, m, and n all have the definitions described in equation (Ib).

[0168] In some embodiments, the antibody conjugate represented by formula (Ic) is as shown in formula (Ic-A): Formula (Ic-A) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

[0169] In some embodiments, the antibody conjugate represented by formula (Ic) is as shown in formula (Ic-B): Formula (Ic-B) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

[0170] In some embodiments, the antibody conjugate shown in formula (I) is as shown in formula (Id): Formula(Id) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof; wherein: DDRi, SP 1 W 1 W 2 HP 1 HP 2 SP 3 RL, Y, L 2 Su, Ab, TOPOi, m, and n all have the definitions described in equation (Ib).

[0171] In some embodiments, the antibody conjugate represented by formula (Id) is as shown in formula (Id-A): Formula (Id-A) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

[0172] In some embodiments, the antibody conjugate represented by formula (Id) is as shown in formula (Id-B): Formula (Id-B) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

[0173] In some implementation schemes, SP 2 and SP 3 These are divalent linking groups that facilitate the introduction of elimination groups, release triggering groups, hydrophobic groups, spacer groups, and / or coupling groups into the compound. Spacer groups, which help separate the coupling group from other groups in the compound, can more effectively couple the compound of the present invention to a second compound and more effectively cleave active catabolic metabolites. Spacer groups can also stabilize the coupling group, thereby improving the overall properties of the antibody-drug conjugate. Useful linking groups are known and obvious to those skilled in the art. Examples of useful linking groups are provided in this invention. In some embodiments, the linking group may comprise a divalent ketone, divalent ester, divalent ether, divalent amide, divalent amine, alkylene, arylene, sulfide, disulfide, carbonylene, or a combination thereof. In some embodiments, the linking group may comprise –C(O)–, –O–, –C(O)NH–, –C(O)NH-alkyl–, –OC(O)NH–, –SC(O)NH–, –NH–, –NH-alkyl–, –C(O)N(CH3)–, –C(O)N(CH3)-alkyl–, –N(CH3)–, –N(CH3)-alkyl–, –N(CH3)CH2CH2N(CH3)–, –C(O)CH2CH2CH2C(O)–, –S–, –SS–, –OCH2CH2O–, or their opposites (e.g., –NHC(O)–), or combinations thereof.

[0174] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), SP 1 and SP 3 Each exists independently, or each is independently selected from -C. 1-6 Alkylene -; -C(O)-; -C 1-6 alkylene-C(O)-, wherein the -C(O)- is respectively connected to W 2 or HP 1 ;-C(O)-C 1-6 Alkylene -C(O)-;-C(O)(C 1-6 alkylene)NR 1 C(O)-;-C(O)(C 1-6 Alkylene)OC(O)-; and -C(O)(C 1-6 (alkylene)SC(O)-; where R 1 It is H or C that is optionally substituted. 1-6 Alkyl groups; and the C 1-6Alkylene, either alone or as part of another group, is optionally substituted with one, two or three substituents selected from halogen, alkyl, haloalkyl, hydroxyl, amino, alkylamino and alkoxy groups, respectively.

[0175] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 2 It does not exist independently, or SP 2 Independently for C 1-6 alkylene-C(O)-, wherein the -C(O)- is attached to W 2 In certain embodiments of formulas (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, SP 2 Independently for C 1-6 Alkylene-C(O)-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 2 Independently for C 1-2 Alkylene-C(O)-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 2 Independently for C 3-4 Alkylene-C(O)-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 2 Independently for C 5-6 Alkylene-C(O)-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 2 Independently -CH2-C(O)-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 2Independently -CH2CH2-C(O)-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 2 Independently -(CH2)5-C(O)-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 2 It does not exist.

[0176] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 3 It does not exist independently, or SP 3 Independently -C(O)-C 1-6 Alkylene-C(O)-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 3 Not present. In certain embodiments of equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), or equation (Ic-B), including any of the foregoing embodiments, SP 3 Independently -C(O)-C 1-6 Alkylene-C(O)-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 3 Independently -C(O)-C 1-4 Alkylene-C(O)-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 3 Independently -C(O)-C4 alkylene-C(O)-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 3 Independently -C(O)(C1-6 alkylene)NR 1C(O)-, where R 1 It is H.

[0177] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 2 It does not exist independently, or SP 2 Independently for C 1-6 alkylene-C(O)- and SP 3 It does not exist independently, or SP 3 Independently -C(O)-C 1-6 Alkylene-C(O)-, where SP 2 The -C(O)- is connected to W 2 In certain embodiments of formulas (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, SP 2 It does not exist independently, or SP 2 Independently for C 1-6 alkylene-C(O)- and SP 3 It does not exist independently, where SP 2 The -C(O)- is connected to W 2 In certain embodiments of formulas (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, SP 2 It does not exist independently, or SP 2 Independently for C 1-6 Alkylene-C(O)-, where SP 2 The -C(O)- is connected to W 2 SP 3 It does not exist independently, or SP 3 Independently -C(O)-C 1-6 alkylene-C(O)-; and SP 1 It does not exist.

[0178] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 1 It does not exist independently, or SP 1 Independently selected from -NR 5 CH2- and -NR 4 -triaryl-CH2-, where R5 It is C 1-6 Alkyl-OCH3, and R 4 Is it H or C? 1-6 Alkyl. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 1 Not present. In certain embodiments of equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), or equation (Ic-B), including any of the foregoing embodiments, SP 1 Independently for -NR 5 CH2-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 1 It is -N(CH2CH2OCH3)CH2-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 1 Independently for -NR 4 -Arylidene-CH2-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 1 Independently -NH-aryl-CH2-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 1 yes ; and each These are the connection points that connect to the rest of the compound.

[0179] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 2 It does not exist independently, or SP 2 Independently for C 1-6 Alkylene-C(O)-, where SP 2 The -C(O)- is connected to W 2 , and SP 1Not present. In certain embodiments of equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), or equation (Ic-B), including any of the foregoing embodiments, SP 2 It does not exist independently, or SP 2 Independently for C 1-6 Alkylene-C(O)-, where SP 2 The -C(O)- is connected to W 2 , and SP 1 Independently selected from -NR 5 CH2- and -NR 4 -Arylene-CH2-.

[0180] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 2 It is a C2 alkylene-C(O)-, and SP 1 It is -N(CH2CH2OCH3)CH2-, where SP 2 The -C(O)- is connected to W 2 In certain embodiments of formulas (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, SP 2 It is a C2 alkylene-C(O)-, and SP 1 yes SP 2 The -C(O)- is connected to W 2 .

[0181] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, SP 2 It does not exist independently, or SP 2 Independently for C 1-6 Alkylene-C(O)-, where SP 2 The -C(O)- is connected to W 2 SP 3 It does not exist independently, or SP 3 Independently -C(O)-C 1-6 alkylene-C(O)-; and SP 1 It does not exist independently, or SP 1 Independently selected from -NR 5CH2- and -NR 4 -Arylene-CH2-.

[0182] In some implementation schemes, W 1 This refers to an eliminator group. Eliminator groups facilitate the separation of the bioactive group portion of the compound or conjugate described in this invention from the rest of the compound or conjugate in vivo and / or in vitro. Eliminator groups can also be used in conjunction with release-initiating groups to further promote the separation of the bioactive group portion of the compound or conjugate described in this invention. For example, an eliminator group and a release-initiating group can undergo a release reaction, thereby releasing the bioactive group portion of the compound or conjugate described in this invention from the compound or conjugate in vivo and / or in vitro. After a release reaction is initiated by a release-initiating group, the eliminator group cleaves the bioactive group portion or its prodrug form, forming a stable, non-toxic entity that does not further affect the activity of the bioactive group portion.

[0183] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 1 Independently In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, W 1 Independently In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, W 1 Independently In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, W 1 Independently .

[0184] In certain embodiments of formulas (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, z is 0. In certain embodiments of formulas (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, z is 1. When z is 1, the conjugate contains a release triggering group (designated RT in this invention), which facilitates the separation of the bioactive group portion of the compound or conjugate of this invention from the remaining portion of the compound or conjugate in vivo and / or in vitro. The release triggering group may also be used in combination with an elimination group to further facilitate the separation of the bioactive group portion of the compound or conjugate of this invention. For example, the elimination group and the release triggering group can react in a release reaction to release the bioactive group portion of the compound or conjugate of the present invention from the compound or conjugate in vivo and / or in vitro. In some embodiments, the release triggering group can function through a biodriven reaction with high tumor:non-tumor specificity (e.g., proteolytic action of an enzyme overexpressed in the tumor microenvironment).

[0185] The hydrophilic groups are designated as HP in this invention. 1 and HP 2 This contributes to improving the hydrophilicity of the compounds described in this invention. It is believed that increased hydrophilicity can enhance the solubility of the compounds in aqueous solutions (e.g., aqueous solutions in biological systems). The hydrophilic group can also serve as a spacer group, which will be described in further detail in this invention. Available hydrophilic groups include those described in this invention. In some embodiments, HP 1 Independently, it is divalent polyethylene glycol. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 1 Independently , where R 2 It is H or methyl, and x1 is an integer between 1 and 50, inclusive; and each These are connection points that connect to the remainder of the compound. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, R 2It is H. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, R 2 It is a methyl group.

[0186] In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, x1 is independently an integer between 1 and 10, inclusive. In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, x1 is independently an integer between 1 and 5, inclusive. In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, x1 is 4.

[0187] In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, x1 is independently an integer between 1 and 10, inclusive; and R 2 It is H. In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, x1 is independently an integer between 1 and 5, inclusive; and R 2 It is H. In certain embodiments of equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), or equation (Ic-B), including any of the foregoing embodiments, x1 is 4; and R 2 It is H.

[0188] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 1 It does not exist.

[0189] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, RT is a β-glucuronide that can be cleaved by β-glucuronidase. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, RT is... , in It is the connection point that connects to the rest of the compound. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, RT is... .

[0190] In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, y is 0. In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, y is 1. In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, y is 2.

[0191] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, L 1 Independently as a key. In certain embodiments of equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), or equation (Ic-B), including any of the foregoing embodiments, L 1 Independently for unreplaced C 1-6 Alkylene. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, L 1 C can be substituted independently. 1-6 alkylene, wherein the C 1-6The alkylene group is optionally substituted by one, two or three substituents selected from halogens, alkyl groups, haloalkyl groups, hydroxyl groups, amino groups, alkylamino groups and alkoxy groups.

[0192] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, R A Each time it appears, it is independently represented as an unsubstituted C. 1-6 Alkyl. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, R A Each time it appears, it is independently methyl. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, R A Each time it appears, it is independently a methyl group, and y is 1. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, R A Each time it appears, it is independently assigned the number C. 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted by one, two or three substituents selected from halogens, alkyl groups, haloalkyl groups, hydroxyl groups, amino groups, alkylamino groups and alkoxy groups.

[0193] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 1 It does not exist independently, or W 1 Independently or In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, W 1 It does not exist independently, or W 1 Independently or In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, W 1 It does not exist independently, or W 1 Independently or In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, W 1 It does not exist. In certain embodiments of equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), or equation (Ic-B), including any of the foregoing embodiments, W 1 yes In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, W 1 yes In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, W 1 yes In certain embodiments of formula (Ia), formula (Ib), formula (Ib-A), or formula (Ib-B), including any of the foregoing embodiments, W 1 yes In some embodiments of equation (Ia), including any of the foregoing embodiments, W 1 yes In certain embodiments of formula (Ia), formula (Ib), formula (Ib-A), or formula (Ib-B), including any of the foregoing embodiments, W 1 yes In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, W 1 yes In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, W 1 yes In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, W 1 yes In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, W 1 yes In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, W 1 It does not exist independently, or W 1 Independently ,or In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, W 1 It does not exist independently, or W 1 Independently , ,or .

[0194] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 It does not exist. In certain embodiments of equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), or equation (Ic-B), including any of the foregoing embodiments, W 2 Independently, it is an amino acid residue or a peptide residue, wherein said amino acid residue or said peptide residue is optionally HP 2 Replacement. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently, it is an unsubstituted amino acid residue. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2Independently, these are amino acid residues, wherein the amino acid residues are HP 2 Replacement. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently, it is an unsubstituted peptide residue. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently, as peptide residues, wherein the peptide residues are HP 2 replace.

[0195] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently, it is a peptide residue, and said peptide residue contains at least one non-natural amino acid. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently, it is a peptide residue, and said peptide residue contains a non-natural amino acid. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently, it is a peptide residue, and said peptide residue contains two non-natural amino acids. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently, the peptide residue comprises at least one non-natural amino acid and the peptide residue is optionally modified by HP. 2 replace.

[0196] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, the at least one non-natural amino acid is selected from 3-sulfoalanine, hydroxyproline (Hyp), citrulline (Cit), ornithine (Orn), normal leucine (Nle), 3-nitrotyrosine, nitroarginine, pyroglutamic acid (Pyr), naphthylalanine (Nal), 2,4-diaminobutyric acid (DAB), methionine sulfoxide, methionine sulfone, and 2,3-diaminopropionic acid. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, the at least one non-natural amino acid is selected from 3-sulfoalanine, citrulline (Cit), 2,3-diaminopropionic acid, and β-alanine.

[0197] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently, these are peptide residues, wherein the peptide residues are optionally converted by HP. 2 The substitutions, and the peptide residues include valine, citrulline (Cit), 3-sulfoalanine, alanine, 2,3-diaminopropionic acid, or β-alanine.

[0198] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, the peptide residue comprises citrulline (Cit). In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, the peptide residue comprises β-alanine. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, the peptide residue comprises 2,3-diaminopropionic acid.

[0199] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, the peptide residues comprise citrulline (Cit) and 3-sulfoalanine. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, the peptide residues comprise β-alanine and 2,3-diaminopropionic acid.

[0200] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, the peptide residues comprise valine and citrulline (Cit). In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, the peptide residues comprise valine, citrulline (Cit), and 3-sulfoalanine. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, the peptide residues comprise valine and alanine. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, the peptide residues comprise valine, citrulline (Cit), and 2,3-diaminopropionic acid. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, the peptide residues comprise β-alanine and 2,3-diaminopropionic acid.

[0201] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently, these are amino acid residues, wherein said amino acid residues are optionally HP 2 Replacement. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2Independently, it is an amino acid residue, and the amino acid residue is a β-amino acid. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 It is β-alanine independently.

[0202] In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, W 2 Independently selected from -(C(O)CHR 3 NR 4 ) a -、-(C(O)CH2CHR 3 NR 4 ) b -、-(C(O)CHR 3 CH2NR 4 ) b -、-(C(O)CH2CHR 3 NR 4 ) b -(C(O)CHR 3 NR 4 ) a -、-(C(O)CHR 3 CH2NR 4 ) b -(C(O)CHR 3 NR 4 ) a -、-(C(O)CHR 3 NR 4 ) a -(C(O)CH2CHR 3 NR 4 ) b -、and-(C(O)CHR 3 NR 4 ) a -(C(O)CHR 3 CH2NR 4 ) b -; Where R 3 Independently, these are amino acid side chain residues, which optionally are HP-dependent. 2 replace; R 4 Independently H or C 1-6 alkyl; a and b are each independent integers between 1 and 10, inclusive; and W 2 The -C(O)- is connected to W 1 .

[0203] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently -(C(O)CHR 3 NR 4 ) a - In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently -(C(O)CHR 3 NH) a -, where a is 1. In certain embodiments of equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), or equation (Ic-B), including any of the foregoing embodiments, W 2 Independently -(C(O)CHR 3 NH) a -, where a is 2. In certain embodiments of equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), or equation (Ic-B), including any of the foregoing embodiments, W 2 Independently -(C(O)CHR 3 NH) a -, where a is 3. In certain embodiments of equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), or equation (Ic-B), including any of the foregoing embodiments, W 2 Independently -(C(O)CHR 3 NR 4 ) a -, and R 3 It is an amino acid side chain residue independently selected from valine, citrulline, alanine, glycine, 3-sulfoalanine, and 2,3-diaminopropionic acid.

[0204] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2Independently -(C(O)CH2CHR 3 NR 4 ) b - In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently -(C(O)CH2CHR 3 NH)-. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 It is independently -(C(O)CH2CH2NH)-.

[0205] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently -(C(O)CH2CHR 3 NR 4 ) b -(C(O)CHR 3 NR 4 ) a - In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently -(C(O)CH2CHR 3 NH) b -(C(O)CHR 3 NH) a - In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently -(C(O)CH2CH2NH)-(C(O)CHR 3 NH) a - In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 Independently -(C(O)CH2CH2NH)-(C(O)CHR 3 NH)-, where R3 It is an amino acid side chain residue independently selected from valine, citrulline, alanine, glycine, 3-sulfoalanine, and 2,3-diaminopropionic acid.

[0206] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 It independently has the structure shown in the following formula: , or ; and each These are the connection points that connect to the rest of the compound.

[0207] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 It independently has the structure shown in the following formula: , or .

[0208] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 It independently has the structure shown in the following formula: .

[0209] In certain embodiments of equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), or equation (Ic-B), including any of the foregoing embodiments, -W 1 -W 2 - Independently possesses the structure shown in the following formula: ,or .

[0210] In certain embodiments of equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), or equation (Ic-B), including any of the foregoing embodiments, -W 1 -W 2 - Independently possesses the structure shown in the following formula: .

[0211] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 It includes polyethylene glycol (PEG), methoxy polyethylene glycol (mPEG), poly(propylene glycol) (PPG), copolymers of ethylene glycol and propylene glycol, poly(oxyethylated polyols), poly(enols), poly(vinylpyrrolidone), poly(hydroxyalkyl methylacrylamide), poly(hydroxyalkyl methacrylates), poly(sugars), poly(α-hydroxy acids), poly(vinyl alcohol), polyphosphazenes, polyoxazoline compounds (POZ), and poly( N -Acryloylmorpholine), polysarcosine, or combinations thereof. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 It is polyethylene glycol (PEG). In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 It is methoxy polyethylene glycol (mPEG). In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 It is poly(propylene glycol) (PPG). In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 It is a copolymer of ethylene glycol and propylene glycol. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 It is a poly(oxyethylated polyol). In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 It is poly(enol). In some implementations, HP 2It is poly(vinylpyrrolidone). In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 It is poly(hydroxyalkylmethylacrylamide). In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 It is poly(hydroxyalkyl methacrylate). In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 It is a polysaccharide. In some implementations, HP 2 It is a poly(α-hydroxy acid). In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 It is poly(vinyl alcohol). In some implementations, HP 2 It is polyphosphazene. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 It is a polyoxazoline (POZ). In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, POLY is a poly( N -Acryloylmorpholine). In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 It is polysarcosine. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 It is a non-peptide water-soluble polymer.

[0212] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 Independently, it is monovalent polyethylene glycol. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, HP 2 Independently , where R 2 It is H or methyl, and x2 is an integer between 1 and 50, inclusive; and each These are connection points that connect to the remainder of the compound. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, R 2 It is H. In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, R 2 It is a methyl group.

[0213] In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, x2 is independently an integer between 1 and 20, inclusive. In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, x2 is independently an integer between 10 and 20, inclusive. In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, x2 is 12.

[0214] In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, x2 is independently an integer between 1 and 20, inclusive; and R 2It is H. In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, x2 is independently an integer between 10 and 20, inclusive; and R 2 It is H. In certain embodiments of equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), or equation (Ic-B), including any of the foregoing embodiments, x2 is 12; and R 2 It is H.

[0215] In certain embodiments of formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), or formula (Ic-B), including any of the foregoing embodiments, W 2 It independently has the structure shown in the following formula: .

[0216] In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, L 2 It is -C 1-6 alkylene-, the -C 1-6 The alkylene group is optionally substituted with one or more substituents selected from the following: halogen, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Haloalkyl. In certain embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, L 2 It is unsubstituted -C 1-6 Alkylene-. In certain embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, L 2 It is -C 1-3 alkylene-, the -C 1-3The alkylene group is optionally substituted with one or more substituents selected from the following: halogen, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Haloalkyl. In certain embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, L 2 It is -CH2-. In some implementations, L 2 It is -CH2CH2-. In certain embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, L 2 It is -CH2CH2CH2-.

[0217] In some embodiments of equations (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, b is 1. In some embodiments of equations (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Y is –X. 1 -C 1-6 Alkylene-[X] 1 -C 1-6 [alkylene] a -X 1 – wherein at least one alkylene group in Y is selected from one or more of R 50 Substituent substitution. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Y is –X. 1 -C 2-6 imide-[X] 1 -C 2-6 [Alkenyl] a -X 1 – where at least one alkenyl group in Y is selected from one or more groups of R50 Substituent substitution. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Y is –X. 1 -C 2-6 Ethyne-[X] 1 -C 2-6 [Iso-ynyl] a -X 1 – where at least one alkyne group in Y is selected from one or more groups from R 50 Substituents are substituted. In some embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, a is 0. In some embodiments, a is 1. In some embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, a is 2. In some embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, a is 3.

[0218] In some embodiments of equations (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, b is 1. In some embodiments of equations (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Y is –X. 1 -C 1-4 Alkylene-[X] 1 -C 1-4 [alkylene] a -X 1 – wherein at least one alkylene group in Y is selected from one or more of R 50Substituents are substituted. In some embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, a is 0. In some embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, a is 1. In some embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, a is 2. In some embodiments, a is 3.

[0219] In some embodiments of equations (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, b is 1. In some embodiments of equations (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Y is –X. 1 -C 1-4 Alkylene-X 1 -C 1-4 Alkylene-X 1 -C 1-4 Alkylene-X 1 -C 1-4 Alkylene-X 1 – wherein at least one alkylene group in Y is selected from one or more of R 50 Substituent substitution. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Y is –X. 1 -C 1-4 Alkylene-X 1 -C 1-4 Alkylene-X 1 -C 1-4 Alkylene-X 1 – wherein at least one alkylene group in Y is selected from one or more of R 50 Substituent substitution. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Y is –X. 1 -C 1-4 Alkylene-X1 -C 1-4 Alkylene-X 1 – wherein at least one alkylene group in Y is selected from one or more of R 50 Substituents are substituted.

[0220] In some embodiments of equations (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, b is 0. In some embodiments of equations (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Y is –X. 1 -C 1-6 Alkylene-[X] 1 -C 1-6 [alkylene] a – wherein at least one alkylene group in Y is selected from one or more of R 50 The substituents in Y are substituted, and the alkylene group in Y is optionally replaced by one or more substituents selected from R. 51 Substituent substitution. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Y is –X. 1 -C 1-6 imide-[X] 1 -C 1-6 [Alkenyl] a – where at least one alkenyl group in Y is selected from one or more groups of R 50 Substituents of Y, and wherein the alkenyl group in Y is optionally replaced by one or more selected from R. 51 Substituent substitution. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Y is –X. 1 -C 2-6 Ethyne-[X] 1 -C 2-6 [Iso-ynyl] a – where at least one alkyne group in Y is selected from one or more groups from R 50 Substituents of Y, and wherein the ynynyl group in Y is optionally replaced by one or more selected from R 51Substituents are substituted. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, a is 0. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, a is 1. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, a is 2. In some embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, a is 3.

[0221] In some embodiments of equations (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, b is 0. In some embodiments of equations (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Y is –X. 1 -C 1-6 Alkylene-[X] 1 -C 1-6 [alkylene] a – wherein at least one alkylene group in Y is selected from one or more of R 50 The substituents in Y are substituted, and the alkylene group in Y is optionally replaced by one or more substituents selected from R. 51 Substituent substitution. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Y is –X. 1 -C 1-4 Alkylene-[X] 1 -C 1-4 [alkylene] a – wherein at least one alkylene group in Y is selected from one or more of R 50 The substituents in Y are substituted, and the alkylene group in Y is optionally replaced by one or more substituents selected from R. 51Substituent substitution. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Y is –X. 1 -C 1-4 Alkylene-X 1 -C 1-4 Alkylene-X 1 -C 1-4 Alkylene – wherein at least one alkylene in Y is selected from one or more of R 50 The substituents in Y are substituted, and the alkylene group in Y is optionally replaced by one or more substituents selected from R. 51 Substituents are substituted. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, a is 0. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, a is 1. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, a is 2. In some embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, a is 3.

[0222] In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, R 51 Independently selected from halogens, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)-. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, R 51 It is a halogen. In certain embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, R 51 It is -CN. In some implementations, R 51It is -NO2. In some embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, R 51 It is -OH. In some implementations, R 51 It is -NH2. In some implementations, R 51 It is -C(O)NH2. In certain embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, R 51 It is -C(O)-.

[0223] In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, X 1 and X 2 Each is independently selected from –N(R) 10 –, –C(O)–, and –N(R)– 10 )C(O)–. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, X 1 and X 2 Each is independently selected from –NH–, –C(O)–, and –N(R)–. 10 )C(O)–. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, X 1 and X 2 Each is independently selected from –C(O)– and –N(R)–. 10 )C(O)–. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, X 1 and X 2 Each is independently selected from –C(O)– and –NHC(O)–.

[0224] In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, R 50Yes –C 1-6 Alkylene-X 2 -[C 1-6 [alkylene] c -HP 2 , where R 50 Each alkylene group is optionally bonded by one or more elements selected from halogens, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Substituents of haloalkyl groups. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, R 50 Yes –C 1-4 Alkylene-X 2 -[C 1-4 [alkylene] c -HP 2 , where R 50 Each alkylene group is optionally bonded by one or more elements selected from halogens, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Substituents of haloalkyl groups. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, R 50 Each alkylene group is optionally bonded by one or more elements selected from halogens, -OH, -N(R) 10 )2、-C(O)N(R 10 )2、-C(O)-、C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C1-10 The alkyl halogroup is substituted. In some embodiments, c is 0. In some embodiments, c is 1.

[0225] In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, R 50 Yes –C 2-6 imide-X 2 -[C 2-6 [Alkenyl] c -HP 2 , where R 50 Each of the alkenyl groups is optionally surrounded by one or more elements selected from halogen, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Substituents of haloalkyl groups. In certain embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, c is 0. In certain embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, c is 1.

[0226] In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, R 50 Yes –C 2-6 Ethyne-X 2 -[C 2-6 [Iso-ynyl] c -HP 2 , where R 50 Each of the alkyne groups is optionally surrounded by one or more groups selected from halogen, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Substituents of haloalkyl groups. In certain embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, c is 0. In certain embodiments of formula (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, c is 1.

[0227] In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, -L 2 -Y- is -C 1-6 Alkylene-NHC(O)-C 1-6 Alkylene-[NHC(O)-C 1-6 [alkylene]2-[C(O)]–, wherein at least one alkylene in Y is selected from one or more of R 50 Substituent substitution. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, -L 2 -Y- is -CH2-NHC(O)-C2 alkylene-NHC(O)-CH2-NHC(O)-C4 alkylene-[C(O)]–, wherein at least one alkylene in Y is selected from one or more of R. 50 Substituent substitution. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, -L 2 -Y- is –CH2-NHC(O)-C2 alkylene-NHC(O)-CH(R) 50 )-NHC(O)-C4 alkylene-[C(O)]–, where R 50 Yes –C 1-6 Alkylene-X 2 -[C 1-6 [alkylene] c -HP 2In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, -L 2 -Y- is –CH2-NHC(O)-C2 alkylene-NHC(O)-CH(R) 50 )-NHC(O)-C4 alkylene-[C(O)]–, where R 50 It is –CH2-NHC(O)-HP 2 .

[0228] In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, -L 2 -Y- is -C 1-6 Alkylene-NHC(O)-C 1-6 Alkylene-[NHC(O)-C 1-6 [alkylene]2–, wherein at least one alkylene in Y is selected from one or more of R 50 Substituent substitution. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, -L 2 -Y- is -CH2-NHC(O)-C2 alkylene-NHC(O)-CH2-NHC(O)-C4 alkylene-, wherein at least one alkylene in Y is selected from one or more of R. 50 Substituent substitution. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, -L 2 -Y- is –CH2-NHC(O)-C2 alkylene-NHC(O)-CH(R) 50 )- NHC(O)- C4 alkylene-, wherein R 50 Yes –C 1-6 Alkylene-X 2 -[C 1-6 [alkylene] c -HP 2 In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, -L 2-Y- is –CH2-NHC(O)-C2 alkylene-NHC(O)-CH(R) 50 )-NHC(O)-C4 alkylene-[C(O)]–, where R 50 It is –CH2-NHC(O)-HP 2 .

[0229] In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Su is a sugar group moiety. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Su is a hexose form of a monosaccharide. Su can be a glucuronic acid residue or a mannose residue. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Su is ,in This indicates a connection point that connects to the remainder of the compound. In certain embodiments of formulas (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, Su is ,in This indicates the connection point that connects to the rest of the compound.

[0230] In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, RL is independently selected from the group consisting of: ;in Each of these is a connection point that connects to the rest of the compound.

[0231] In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, RL is or In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, RL is In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, RL is In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, RL is In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, RL is In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, RL is .

[0232] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the topoisomerase inhibitor is a topoisomerase I inhibitor. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the topoisomerase I inhibitor is a camptothecin derivative. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the topoisomerase I inhibitor is independently selected from camptothecin, irinotecan, SN-38, topotecan, and exatecan. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the topoisomerase I inhibitor is exatecan.

[0233] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the topoisomerase inhibitor is independently selected from the group consisting of: ; or its derivatives; and each These are the connection points that connect to the rest of the compound.

[0234] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the topoisomerase inhibitor is independently selected from the group consisting of: ,and ; or its derivatives.

[0235] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the topoisomerase inhibitor is independently selected from the group consisting of: ; or its derivatives.

[0236] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the topoisomerase inhibitor is a topoisomerase II inhibitor. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the topoisomerase II inhibitor is independently selected from etoposide, teniposide, and tafluposide.

[0237] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is a poly(ADP-ribose) polymerase (PARP) inhibitor, a checkpoint kinase 1 (CHK1) inhibitor, an ataxia-telangiectasia and Rad3-associated (ATR) kinase inhibitor, a nicotinamide phosphoribosyltransferase 1 (NAMPT1) inhibitor, or a RAD51 inhibitor.

[0238] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is a poly(ADP-ribose) polymerase (PARP) inhibitor. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is a checkpoint kinase 1 (CHK1) inhibitor. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is a checkpoint kinase 2 (CHK2) inhibitor. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is a checkpoint kinase 1 / 2 (CHK1 / 2) inhibitor. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is an ataxia-telangiectasia and Rad3-associated (ATR) kinase inhibitor. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is an ataxia-telangiectasia mutant (ATM) kinase inhibitor. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is a nicotinamide phosphoribosyltransferase 1 (NAMPT1) inhibitor.In some embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is a RAD51 inhibitor.

[0239] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is a poly(ADP-ribose) polymerase (PARP) inhibitor, and the PARP inhibitor is independently selected from olaparib, veliparib, talazoparib, rucaparib, niraparib, AZD5305, talzenna, and derivatives thereof. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the PARP inhibitor is olaparib or a derivative thereof. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the PARP inhibitor is veliparib or a derivative thereof. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the PARP inhibitor is talazoparib or a derivative thereof. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the PARP inhibitor is rucaparib or a derivative thereof. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the PARP inhibitor is niraparib or a derivative thereof.In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the PARP inhibitor is AZD5305 or a derivative thereof. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the PARP inhibitor is talzenna or a derivative thereof.

[0240] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the PARP inhibitor is independently selected from... or its derivatives; and each These are the connection points that connect to the rest of the compound.

[0241] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the PARP inhibitor is independently selected from... ; or its derivatives; and each These are the connection points that connect to the rest of the compound.

[0242] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the PARP inhibitor is independently selected from... ; or its derivatives; and each These are the connection points that connect to the rest of the compound.

[0243] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the PARP inhibitor is independently selected from... ,and ; or its derivatives.

[0244] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the PARP inhibitor is or its derivatives. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the PARP inhibitor is or its derivatives.

[0245] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is an independently selected CHK1 inhibitor of 7-hydroxystaurosporine, AZD7762, rabusertib, MK-8776, prexasertib, GDC-575, and CCT245737.

[0246] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is independently selected from the following CHK1 inhibitors: ; or its derivatives; and each These are the connection points that connect to the rest of the compound.

[0247] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is independently selected from the following CHK1 inhibitors: ; or its derivatives.

[0248] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is independently selected from the following CHK1 inhibitors: ,and ; or its derivatives.

[0249] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is or its derivatives.

[0250] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the DDR inhibitor is an ataxia-telangiectasia and Rad3-associated (ATR) kinase inhibitor, wherein the ataxia-telangiectasia and Rad3-associated (ATR) kinase inhibitor is independently selected from bezosertib, ceralasertib, elimusertib, M4344 (VX-803), BAY1895344, schisandrin B, etc. B), NU6027, dactolisib, EP-46464, Torin-2, VE-821, AZ20, IMP9064, ATG-018, ATRN-119, and ART0380.

[0251] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the ATR inhibitor is independently selected from... ; or its derivatives; and each These are the connection points that connect to the rest of the compound.

[0252] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the ATR inhibitor is independently selected from... ; or its derivatives; and each These are the connection points that connect to the rest of the compound.

[0253] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the ATR inhibitor is or its derivatives.

[0254] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the nicotinamide phosphoribosyltransferase 1 (NAMPT1) inhibitor is independently selected from GMX1777, FK866, CHS828, OT-82, APO866, KPT-9274, and ATG-019.

[0255] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the NAMPT1 inhibitor is independently selected from... , ; or its derivatives; and each These are the connection points that connect to the rest of the compound.

[0256] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the NAMPT1 inhibitor is independently selected from... , ; or its derivatives.

[0257] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the NAMPT1 inhibitor is independently selected from... ,and , or its derivatives.

[0258] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the NAMPT1 inhibitor is independently selected from... ,and , or its derivatives.

[0259] In certain embodiments of equations (I), (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, m and n are each independently integers between 1 and 10, inclusive. In certain embodiments of equations (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, m and n are each independently integers between 1 and 8, inclusive. In certain embodiments of equations (I), (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, m and n are each independently integers between 1 and 5, inclusive of 1 and 5. In certain embodiments of equations (I), (Ia-1), (Ia), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, m and n are each independently integers between 1 and 4, inclusive of 1 and 4.

[0260] In certain embodiments of equation (I), equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), equation (Ic-B), equation (Id), equation (Id-A), or equation (Id-B), including any of the foregoing embodiments, m is 2 and n is 4. In certain embodiments of equation (I), equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), equation (Ic-B), equation (Id), equation (Id-A), or equation (Id-B), including any of the foregoing embodiments, m is 4 and n is 4. In certain embodiments of equation (I), equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), equation (Ic-B), equation (Id), equation (Id-A), or equation (Id-B), including any of the foregoing embodiments, m is 8 and n is 4. In certain embodiments of equation (I), equation (Ia-1), equation (Ia), equation (Ib), equation (Ib-A), equation (Ib-B), equation (Ic), equation (Ic-A), equation (Ic-B), equation (Id), equation (Id-A), or equation (Id-B), including any of the foregoing embodiments, m is 8 and n is 8.

[0261] In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the molar ratio of m to n is 1:1. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the molar ratio of m to n is 2:1. In certain embodiments of formula (I), formula (Ia-1), formula (Ia), formula (Ib), formula (Ib-A), formula (Ib-B), formula (Ic), formula (Ic-A), formula (Ic-B), formula (Id), formula (Id-A), or formula (Id-B), including any of the foregoing embodiments, the molar ratio of m to n is 1:2.

[0262] In some embodiments of equations (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, yes ,in This indicates a connection point connected to the remainder of the general formula. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, yes In some embodiments of equations (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, yes In some embodiments of equations (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, yes In some embodiments of equations (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, yes In some embodiments of equations (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, yes In some embodiments of equations (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, yes In some embodiments of equations (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, yes .

[0263] In some embodiments of formulas (Ia), (Ia-1), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, yes .

[0264] In some embodiments of equations (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, Choose from the following groups: SP 1 SP 3 ,RL,R 3 x1, a, b, and m are all as defined in this invention; Indicates the connection point connected to DDRi; and This indicates the connection point that connects to the rest of the general formula.

[0265] In some embodiments of equations (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, Choose from the following groups: SP 1 SP 3 ,RL,R 3 x1, a, b, and m are all as defined in this invention; Indicates the connection point connected to DDRi; and This indicates the connection point that connects to the rest of the general formula.

[0266] In some embodiments of equations (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, Choose from the following groups: .

[0267] In some embodiments of formulas (Ia), (Ia-1), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, yes SP 1 SP 3 ,RL,R 3 a, b, and m are all as defined in this invention; Indicates the connection point to TOPOi; and This indicates the connection point that connects to the rest of the general formula.

[0268] In some embodiments of formulas (Ia), (Ia-1), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, yes or ; Or a mixture thereof.

[0269] In some embodiments of equations (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, Choose from the following groups: .

[0270] In some embodiments of equations (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, Choose from the following groups: and .

[0271] In some embodiments of equations (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, Choose from the following groups: .

[0272] In some embodiments of equations (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, yes .

[0273] In some embodiments of formulas (Ia), (Ia-1), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, yes or ; Or a mixture thereof.

[0274] In some embodiments of formula (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, yes or ; Or a mixture thereof.

[0275] In some embodiments of formula (Ia) or (Ia-1), including any of the foregoing embodiments, yes or ; or mixtures thereof; and yes .

[0276] In some embodiments of formula (Ia) or (Ia-1), including any of the foregoing embodiments, yes or ; or mixtures thereof; and yes .

[0277] In some embodiments of formula (Ia) or (Ia-1), including any of the foregoing embodiments, yes or ; or mixtures thereof; and yes .

[0278] In some embodiments of formula (Ia) or (Ia-1), including any of the foregoing embodiments, yes or ; or mixtures thereof; and yes .

[0279] In some embodiments of formula (Ia) or (Ia-1), including any of the foregoing embodiments, yes or ; or mixtures thereof; and yes .

[0280] In some embodiments of formula (Ia) or (Ia-1), including any of the foregoing embodiments, yes or mixtures thereof; and yes .

[0281] In some embodiments of formula (Ia) or (Ia-1), including any of the foregoing embodiments, yes or mixtures thereof; and yes .

[0282] In some embodiments of formula (Ia) or (Ia-1), including any of the foregoing embodiments, yes or mixtures thereof; and yes .

[0283] In some embodiments of formula (Ia) or (Ia-1), including any of the foregoing embodiments, yes or mixtures thereof; and yes .

[0284] In some embodiments of formula (Ia) or (Ia-1), including any of the foregoing embodiments, yes or mixtures thereof; and yes .

[0285] In some embodiments of formula (Ia) or (Ia-1), including any of the foregoing embodiments, yes or mixtures thereof; and yes .

[0286] In some embodiments of formula (Id), (Id-A), or (Id-B), including any of the foregoing embodiments, yes or mixtures thereof; and yes .

[0287] Other connector payloads and couplings are described in PCT / US23 / 26338, entitled “β-GLUCURONIDE LINKER-PAYLOADS, PROTEIN CONJUGATES THEREOF, AND METHODS THEREOF”, the entire contents of which are incorporated herein by reference for all purposes.

[0288] In some embodiments of equations (Ia), (Ia-1), (Ib), (Ib-A), or (Ib-B), including any of the foregoing embodiments, Choose the group consisting of the following equations 1101-1114 and 1115-1118: .

[0289] In some embodiments of formulas (Ia), (Ia-1), (Ic), (Ic-A), or (Ic-B), including any of the foregoing embodiments, It is formula 1201A or 1201B, or a mixture thereof: .

[0290] In some embodiments of formula (Ia) or (Ia-1), including any of the foregoing embodiments, or It binds to p-azidomethyl-L-phenylalanine (pAMF) at one or more sites, and and Another one binds to p-azidomethyl-L-phenylalanine (pAMF) at one or more sites.

[0291] In some embodiments of formula (Ia) or (Ia-1), including any of the foregoing embodiments, It binds to p-azidomethyl-L-phenylalanine (pAMF) at one or more sites, and It binds to p-azidomethyl-L-phenylalanine (pAMF) at one or more sites. In some embodiments of formula (Ia) or (Ia-1), including any of the foregoing embodiments, It binds to p-azidomethyl-L-phenylalanine (pAMF) at HC-F241 and HC-404, and It binds to p-azidomethyl-L-phenylalanine (pAMF) at LC-K42.

[0292] In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1101. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1102. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1103. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1104. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1105. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1106. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1107. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1108. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1109. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1110.In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1111. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1112. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1113. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1114. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1115. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1116. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1117. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1118.

[0293] In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1201A or 1201B, or a mixture thereof.

[0294] In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1101 and formula 1201A or 1201B, or a mixture thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1102 and formula 1201A or 1201B, or a mixture thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1103 and formula 1201A or 1201B, or a mixture thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1104 and formula 1201A or 1201B, or a mixture thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1105 and formula 1201A or 1201B, or a mixture thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1106 and formula 1201A or 1201B, or a mixture thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1107 and formula 1201A or 1201B, or a mixture thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1108 and formula 1201A or 1201B, or a mixture thereof.In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1109 and formula 1201A or 1201B, or a mixture thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1110 and formula 1201A or 1201B, or a mixture thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1111 and formulas 1201A or 1201B, or mixtures thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1112 and formulas 1201A or 1201B, or mixtures thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1113 and formula 1201A or 1201B, or a mixture thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1114 and formula 1201A or 1201B, or a mixture thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1115 and formula 1201A or 1201B, or a mixture thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1116 and formula 1201A or 1201B, or a mixture thereof.In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1115 and formula 1201A or 1201B, or a mixture thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1117 and formula 1201A or 1201B, or a mixture thereof. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1118 and formulas 1201A or 1201B or a mixture thereof.

[0295] In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1101 and formulas 1201A or 1201B, or mixtures thereof, wherein m is 2 and n is 4. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1102 and formulas 1201A or 1201B, or mixtures thereof, wherein m is 2 and n is 4. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1103 and formulas 1201A or 1201B, or mixtures thereof, wherein m is 2 and n is 4. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1104 and formulas 1201A or 1201B, or mixtures thereof, wherein m is 2 and n is 4. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1105 and formulas 1201A or 1201B or mixtures thereof, wherein m is 2 and n is 4. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1106 and formulas 1201A or 1201B or mixtures thereof, wherein m is 2 and n is 4. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1107 and formulas 1201A or 1201B, or mixtures thereof, wherein m is 2 and n is 4. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1108 and formulas 1201A or 1201B, or mixtures thereof, wherein m is 2 and n is 4.In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1109 and formulas 1201A or 1201B, or mixtures thereof, wherein m is 2 and n is 4. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1110 and formulas 1201A or 1201B, or mixtures thereof, wherein m is 2 and n is 4. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1111 and formulas 1201A or 1201B, or mixtures thereof, wherein m is 2 and n is 4. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1112 and formulas 1201A or 1201B, or mixtures thereof, wherein m is 2 and n is 4. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1113 and formulas 1201A or 1201B, or mixtures thereof, wherein m is 2 and n is 4. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises formula 1114 and formulas 1201A or 1201B, or mixtures thereof, wherein m is 2 and n is 4.

[0296] In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises: (a) a linker payload selected from formulas 1101–1118, and (b) a linker payload selected from formulas 1201A or 1201B or mixtures thereof, wherein m is 4 and n is 4. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises: (a) a linker payload selected from formulas 1101–1118, and (b) a linker payload selected from formulas 1201A or 1201B or mixtures thereof, wherein m is 8 and n is 4. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises: (a) a linker payload selected from formulas 1101–1118, and (b) a linker payload selected from formulas 1201A or 1201B or mixtures thereof, wherein m + n is at least 8. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises: (a) a linker payload selected from formulas 1101–1118, and (b) a linker payload selected from formulas 1201A or 1201B or mixtures thereof, wherein m+n is at least 10. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises: (a) a linker payload selected from formulas 1101–1118, and (b) a linker payload selected from formulas 1201A or 1201B or mixtures thereof, wherein m + n is at least 12. In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises: (a) a linker payload selected from formulas 1101–1118, and (b) a linker payload selected from formulas 1201A or 1201B or mixtures thereof, wherein m + n is at least 14.

[0297] In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises a structure selected from formulas 1101, 1102, 1103, 1104, 1105, 1106, 1107, 1108, 1109, 1110, 1111, 1112, 1113, 1114, 1115, 1116, 1117, and 1118, wherein the structure binds to p-azidomethyl-L-phenylalanine (pAMF) at one or more sites.

[0298] In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises a structure selected from formula 1201A or formula 1201B or a mixture thereof, wherein the structure binds to p-azidomethyl-L-phenylalanine (pAMF) at one or more sites.

[0299] In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises: (a) a product selected from formulas 1101, 1102, 1103, 1104, 1105, 1106, 1107, 1108, 1109, ... (a) Structures of formulas 1110, 1111, 1112, 1113, 1114, 1115, 1116, 1117, and 1118, wherein said structure binds to p-azidomethyl-L-phenylalanine (pAMF) at one or more sites; and (b) a structure selected from formula 1201A or formula 1201B or a mixture thereof, wherein said structure binds to p-azidomethyl-L-phenylalanine (pAMF).

[0300] In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises: (a) selected from formulas 1101, 1102, 1103, 1104, 1105, 1106, 1107, 1108, 1109, 1110, and 1101. 111, the structures of formulas 1112, 1113, 1114, 1115, 1116, 1117 and 1118, wherein the structures are bound to p-azidomethyl-L-phenylalanine (pAMF) at HC-F241 and HC-404; and (b) the structures selected from formula 1201A or formula 1201B or mixtures thereof, wherein the structures are bound to p-azidomethyl-L-phenylalanine (pAMF) at LC-K42.

[0301] In some embodiments of formulas (Ia), (Ia-1), (Ib), (Ib-A), (Ib-B), (Ic), (Ic-A), (Ic-B), (Id), (Id-A), or (Id-B), the antibody conjugate comprises: (a) selected from formulas 1101, 1102, 1103, 1104, 1105, 1106, 1107, 1108, 1109, and 11... 10. Structures of formulas 1111, 1112, 1113, 1114, 1115, 1116, 1117, and 1118, wherein the structures are bonded to p-azidomethyl-L-phenylalanine (pAMF) at HC-F241 and HC-404; and (b) structures selected from formula 1201A or formula 1201B or mixtures thereof, wherein m and n are each independently integers between 1 and 8, including 1 and 8.

[0302] In some embodiments, the antibody comprises a light chain. In some states, the light chain is a κ light chain. In some states, the light chain is a λ light chain.

[0303] In some embodiments, the antibody comprises a heavy chain. In some samples, the heavy chain is IgA. In some samples, the heavy chain is IgD. In some samples, the heavy chain is IgE. In some samples, the heavy chain is IgG. In some samples, the heavy chain is IgM. In some samples, the heavy chain is IgG1. In some samples, the heavy chain is IgG2. In some samples, the heavy chain is IgG3. In some samples, the heavy chain is IgG4. In some samples, the heavy chain is IgA1. In some samples, the heavy chain is IgA2.

[0304] In some embodiments, the antibody is an antibody fragment. In some samples, the antibody fragment is an Fv fragment. In some samples, the antibody fragment is a Fab fragment. In some samples, the antibody fragment is an F(ab′)2 fragment. In some samples, the antibody fragment is a Fab′ fragment. In some samples, the antibody fragment is an scFv (sFv) fragment. In some samples, the antibody fragment is an scFv-Fc fragment.

[0305] In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a polyclonal antibody.

[0306] In some embodiments, the antibody is a chimeric antibody. In some embodiments, the antibody is a humanized antibody. In some embodiments, the antibody is a human antibody.

[0307] In some embodiments, the antibody is an affinity-matured antibody. In some embodiments, the antibody is an affinity-matured antibody derived from the exemplary sequences provided in this invention.

[0308] In some embodiments, the antibody is an anti-tissue factor antibody. In some embodiments, the antibody comprises three heavy chain CDRs of SEQ ID NO: 2752 and three light chain CDRs of SEQ ID NO: 3061, comprising a light chain variable region. In some embodiments, the antibody comprises a heavy chain variable region according to SEQ ID NO: 2752 and a light chain variable region according to SEQ ID NO: 3061. In some embodiments, the antibody comprises heavy chain CDRs according to SEQ ID NO: 130 or SEQ ID NO: 455, SEQ ID NO: 780 or SEQ ID NO: 1105 and SEQ ID NO: 1430 or SEQ ID NO: 1755 and light chain CDRs according to SEQ ID NO: 299, SEQ ID NO: 302 and SEQ ID NO: 305.

[0309] In some embodiments, the antibody is an anti-folate antibody. In some embodiments, the antibody is as described in the following literature: U.S. Patent No. 10,596,270, issued March 24, 2020, entitled "Anti-folate receptor antibody conjugates, compositions comprising anti-folate receptor antibody conjugates, and methods of making and using anti-folate receptor antibody conjugates," the entire contents of which are incorporated herein by reference. In some embodiments, the antibody is identified as antibody 1848-H01 as described in U.S. Patent No. 10,596,270. In some embodiments, the antibody comprises three heavy chain CDRs of the heavy chain variable region SEQ ID NO: 362 as described in U.S. Patent No. 10,596,270 and three light chain CDRs comprising the light chain variable region SEQ ID NO: 367. In some embodiments, the antibody comprises the heavy chain variable region according to SEQ ID NO: 362 as described in U.S. Patent No. US10,596,270, and the light chain variable region according to SEQ ID NO: 367. In some embodiments, the antibody comprises the heavy chain CDR according to SEQ ID NO: 117, SEQ ID NO: 235, and SEQ ID NO: 294 as described in U.S. Patent No. US10,596,270, and the light chain CDR according to SEQ ID NO: 299, SEQ ID NO: 302, and SEQ ID NO: 305. In some embodiments, the antibody comprises the heavy chain CDR according to SEQ ID NO: 58, SEQ ID NO: 176, and SEQ ID NO: 294 as described in U.S. Patent No. US10,596,270, and the light chain CDR according to SEQ ID NO: 299, SEQ ID NO: 302, and SEQ ID NO: 305.

[0310] In some embodiments, the antibody is an anti-HER2 antibody. In some embodiments, the antibody is trastuzumab. In some embodiments, the antibody comprises three heavy chain CDRs of SEQ ID NO: 362 and three light chain CDRs of SEQ ID NO: 402, comprising a heavy chain variable region. In some embodiments, the antibody comprises a heavy chain variable region according to SEQ ID NO: 403 and a light chain variable region according to SEQ ID NO: 367. In some embodiments, the antibody comprises the heavy chain CDRs of SEQ ID NO: 400, SEQ ID NO: 401 and SEQ ID NO: 402 provided in this invention, and the light chain CDRs of SEQ ID NO: 299, SEQ ID NO: 302 and SEQ ID NO: 305 as described in U.S. Patent No. US10,596,270.

[0311] For any of the foregoing embodiments, the scope of consideration for the embodiments of the present invention also includes antibody-drug conjugates, wherein the antibody is selected from various therapeutic antibodies that have been approved for clinical trials or are under development for clinical use. In some embodiments, the antibody comprises three heavy chain CDRs and three light chain CDRs of a therapeutic antibody. In some embodiments, the antibody comprises a heavy chain variable region and a light chain variable region of a therapeutic antibody. In some embodiments, the antibody comprises a heavy chain and a light chain of a therapeutic antibody. In some embodiments, the antibody further comprises one or more amino acid mutations. For example, in some embodiments, the antibody comprises one or more amino acid substitutions having modified amino acids to facilitate linkage with the linker-payload described in the present invention. Exemplary therapeutic antibodies include, but are not limited to, rituximab (Rituxan®, IDEC / Genentech / Roche) (see, for example, US Patent No. 5,736,137), a chimeric anti-CD20 antibody approved for the treatment of non-Hodgkin's lymphoma; HuMax-CD20, an anti-CD20 currently being developed by Genmab, an anti-CD20 antibody described in US Patent No. 5,500,362, AME-133 (Applied Molecular Evolution), and hA20 (Immunomedics, Inc.), HumaLYM (Intracel), and PRO70769 (PCT application number PCT / US2003 / 040426); trastuzumab (Herceptin®, Genentech) (see, for example, US Patent No. US5,677,171), a humanized anti-Her2 / neu antibody approved for the treatment of breast cancer; pertuzumab (rhuMab-2C4, Omnitarg®), currently being developed by Genentech; anti-Her2 antibody (US Patent No. US4,753,894); cetuximab (Erbitux®, Imclone) (US Patent No. US4,943,533; PCT Publication No. WO 96 / 40210), a chimeric anti-EGFR antibody in clinical trials for various cancers; ABX-EGF (US Patent No. US6,235,883), currently being developed by Abgenix-Immunex-Amgen; HuMax-EGFR (US Patent No. US7,247,301), currently being developed by Genmab; 425, EMD55900, EMD62000 and EMD72000 (Merck KGaA) (US Patent No. US5,558,864; Murthy, et al. (1987) Arch. Biochem. Biophys. 252(2): 549-60;Rodeck, et al. (1987) J. Cell. Biochem. 35(4): 315-20; Kettleborough, et al. (1991) Protein Eng. 4(7):773-83); ICR62 (Institute of Cancer Research) (PCT Publication No. WO 95 / 20045; Modjtahedi, et al. (1993) J. Cell. Biophys. 22(I-3): 129-46; Modjtahedi, et al. (1993) Br. J. Cancer 67(2): 247-53; Modjtahedi, et al. (1996) Br. J. Cancer 73(2): 228-35; Modjtahedi, et al. (2003) Int. 105(2): 273-80); TheraCIMhR3 (YMBiosciences, Canada; and Centro de Immunologia Molecular, Cuba) (US Patent No. US5,891,996; US Patent No. US6,506,883; Mateo, et al. (1997) Immunotechnol. 3(1): 71-81); mAb-806 (Ludwig Institue for Cancer Research, Memorial Sloan-Kettering) (Jungbluth, et al. (2003) Proc. Natl. Acad. Sci. USA. 100(2): 639-44); KSB-102 (KSBiomedix); MR1-1 (IVAX, National Cancer Institute) (PCT Publication No. WO 01 / 62931A2); and SC100 (Scancell) (PCT Publication No. WO 01 / 88138); alemtuzumab (Campath®, Millenium), currently approved for the treatment of B-cell chronic lymphocytic leukemia; moromumab CD3 (Orthoclone OKT3®), an anti-CD3 antibody developed by Ortho Biotech / Johnson & Johnson; teimumab (Zevalin®), an anti-CD20 antibody developed by IDEC / Schering AG;Gelatinumab / Ozolmicin (Mylotarg®), an anti-CD33 (P67 protein) antibody developed by Celltech / Wyeth; Amevive®, an anti-LFA-3Fc fusion protein developed by Biogen; ReoPro®, abciximab developed by Centocor / Eliley; Simulect®, baliximab developed by Novartis; Synagis®, palizumab developed by Medimmune; Remicade®, an anti-TNFα antibody developed by Centocor; Humira®, an anti-TNFα antibody developed by Abbott; Humicade®, an anti-TNFα antibody developed by Celltech; Golimumab (CNTO-148), a fully human TNF antibody developed by Centocor; Enbrel®, p75 developed by Immunex / Amgen. TNF receptor Fc fusion protein; Ienercept, a p55TNF receptor Fc fusion protein previously developed by Roche; ABX-CBL, an anti-CD147 antibody being developed by Abgenix; ABX-IL8, an anti-IL8 antibody being developed by Abgenix; ABX-MA1, an anti-MUC18 antibody being developed by Abgenix; Pemtumomab (R1549, 90Y-muHMFG1), an anti-MUC1 antibody being developed by Antisoma; Therex (R1550), an anti-MUC1 antibody being developed by Antisoma; AngioMab (AS1405), being developed by Antisoma; HuBC-1, being developed by Antisoma; Thioplatin (AS1407), being developed by Antisoma; Antegren® (natezumab), an anti-α-4-β-1 (VLA-4) and α-4-β-7 antibody being developed by Biogen; VLA-1 mAb, an anti-VLA-1 integrin antibody being developed by Biogen; LTBR mAb, an anti-lymphotoxin β receptor (LTBR) antibody being developed by Biogen; CAT-152, an anti-TGF-β antibody being developed by Cambridge Antibody Technology; ABT 874 (J695), an anti-IL-12 p40 antibody being developed by Abbott; CAT-192, an anti-TGFβ1 antibody being developed by Cambridge Antibody Technology and Genzyme; CAT-213, an anti-Eotaxin1 antibody being developed by Cambridge Antibody Technology.LymphoStat-B®, an anti-Blys antibody being developed by Cambridge Antibody Technology and Human Genome Sciences Inc.; TRAIL-R1 mAb, an anti-TRAIL-R1 antibody being developed by Cambridge Antibody Technology and Human Genome Sciences Inc.; Avastin® bevacizumab, rhuMAb-VEGF, an anti-VEGF antibody being developed by Genentech; an anti-HER receptor family antibody being developed by Genentech; anti-tissue factor (ATF), an anti-tissue factor antibody being developed by Genentech; Xolair® (omalizumab), an anti-IgE antibody being developed by Genentech; Raptiva® (efalizumab), an anti-CD11a antibody being developed by Genentech and Xoma; MLN-02 antibody (formerly LDP-02), being developed by Genentech and Millenium Pharmaceuticals; HuMax CD4, an anti-CD4 antibody being developed by Genmab; HuMax-IL15, an anti-IL15 antibody being developed by Genmab and Amgen; HuMax-Inflam, being developed by Genmab and Medarex; HuMax-Cancer, an anti-heparinase I antibody being developed by Genmab, Medarex, and Oxford GcoSciences; HuMax-Lymphoma, being developed by Genmab and Amgen; HuMax-TAC, being developed by Genmab; IDEC-131, an anti-CD40L antibody being developed by IDEC Pharmaceuticals; IDEC-151 (Clenoliximab), an anti-CD4 antibody being developed by IDEC Pharmaceuticals; IDEC-114, an anti-CD80 antibody being developed by IDEC Pharmaceuticals; IDEC-152, an anti-CD23 antibody being developed by IDEC Pharmaceuticals; and more. The following are also mentioned: an anti-macrophage migration factor (MIF) antibody developed by Pharmaceuticals; BEC2, an anti-idiotype antibody being developed by Imclone; IMC-1C11, an anti-KDR antibody being developed by Imclone; DC101, an anti-flk-1 antibody being developed by Imclone; and an anti-VE cadherin antibody being developed by Imclone.CEA-Cide® (Iabetuzumab), an anti-carcinoembryonic antigen (CEA) antibody being developed by Immunomedics; LymphoCide® (epazolizumab), an anti-CD22 antibody being developed by Immunomedics; AFP-Cide being developed by Immunomedics; MyelomaCide being developed by Immunomedics; LkoCide being developed by Immunomedics; ProstaCide being developed by Immunomedics; MDX-010, an anti-CTLA4 antibody being developed by Medarex; MDX-060, an anti-CD30 antibody being developed by Medarex; MDX-070 being developed by Medarex; MDX-018 being developed by Medarex; Osidem® (IDM-1), a antibody being developed by Medarex and Immuno-Designed... Molecules developed an anti-Her2 antibody; HuMax®-CD4, an anti-CD4 antibody being developed by Medarex and Genmab; HuMax-IL15, an anti-IL15 antibody being developed by Medarex and Genmab; CNTO 148, an anti-TNFα antibody being developed by Medarex and Centocor / J&J; CNTO 1275, an anti-cytokine antibody being developed by Centocor / J&J; MOR101 and MOR102, anti-intercellular adhesion molecule-1 (ICAM-1) (CD54) antibodies being developed by MorphoSys; MOR201, an anti-fibroblast growth factor receptor 3 (FGFR-3) antibody being developed by MorphoSys; Nuvion® (vesizumab), an anti-CD3 antibody being developed by Protein Design Labs; HuZAF®, an anti-CD3 antibody being developed by Protein Design Labs. The following are described: an anti-gamma interferon antibody developed by ProteinDesign Labs; an anti-α5β1 interferon antibody being developed by ProteinDesign Labs; an anti-IL-12 antibody being developed by ProteinDesign Labs; ING-1, an anti-Ep-CAM antibody being developed by Xoma; Xolair® (omalizumab), a humanized anti-IgE antibody being developed by Genentech and Novartis; and MLN01, an anti-β2 integrin antibody being developed by Xoma. In another embodiment, the therapeutic agents include KRN330 (Kirin); huA33 antibody (A33, Ludwig Cancer Institute); CNTO 95 (αV integrin, Centocor); and MEDI-522 (αVβ3 integrin, Medimmune).Voloximab (αVβ1 integrin, Biogen / PDL); Human mAb216 (B cell glycosylated antigenic epitope, NCL); BiTEMT103 (bispecific CD19×CD3, MedImmune); 4G7×H22 (bispecific B cell×FcγR1, Medarex / Merck KGa); rM28 (bispecific CD28×MAPG, European Patent No. EP1444268); MDX447 (EMD 82633) (bispecific CD64×EGFR, Medarex); Caputoxumab (removab) (bispecific EpCAM×anti-CD3, Trion / Fres); Ertuxomab (bispecific HER2 / CD3, Fresenius Biotech); Oregomab (OvaRex) (CA-125, ViRexx); Rencarex® (WX G250) (carbonic anhydrase IX, Wiex); CNTO 888 (CCL2, Centocor); TRC105 (CD105 (endothelial glycoprotein), Tracon); BMS-663513 (CD137 agonist, Brystol Myers Squibb); MDX-1342 (CD19, Medarex); Siplizumab (MEDI-507) (CD2, Medimmune); Humax-CD20 (CD20, Genmab); Rituximab (CD20, Genentech); Veltuzumab (hA20) (CD20, Immunomedics); Iprazumab (CD22, Amgen); Luxizumab (IDEC 152) (CD23, Biogen); Moromumab-CD3 (CD3, Ortho); HuM291 (CD3 fc receptor, PDL) Biopharma); HeFi-1 (CD30, NCI); MDX-060 (CD30, Medarex); MDX-1401 (CD30, Medarex); SGN-30 (CD30, Seattle Genentics); SGN-33 (Lintozumab) (CD33, Seattle Genentics); Zanomumab (HuMax-CD4) (CD4, Genmab); HCD122 (CD40, Novartis); SGN-40 (CD40, Seattle Genentics); Camppath 1h (Alemtuzumab) (CD52, Genzyme); MDX-1411 (CD70, Medarex);hLL1 (EPB-1) (CD74.38, Immunomedics); Galiximab (IDEC-144) (CD80, Biogen); MT293 (TRC093 / D93) (cleaved collagen, Tracon); HuLuc63 (CS1, PDLPharma); ipilimumab (MDX-010) (CTLA4, Brystol MyersSquibb); Tremelimumab (Ticilimumab, CP-675,2) (CTLA4, Pfizer); HGS-ETR1 (mapamumumab) (DR4TRAIL-R1 agonist, Human Genome Science / Glaxo Smith Kline); AMG-655 (DR5, Amgen); Apomab (DR5, Genentech); CS-1008 (DR5, Daiichi Sankyo); HGS-ETR2 (Lesamumab) (DR5TRAIL-R2 agonist, HGS); Cetuximab (Erbitux) (EGFR, Imclone); IMC-11F8 (EGFR, Imclone); Nimotuzumab (EGFR, YM) Bio); Panitumumab (Vectabix) (EGFR, Amgen); Zaltumumab (HuMaxEGFr) (EGFR, Genmab); CDX-110 (EGFRvIII, AVANTImmunotherapeutics); Adecatumumab (MT201) (Epcam, Merck); Panorex (17-1A) (Epcam, Glaxo / Centocor); MORAb-003 (Folic Acid Receptor α, Morphotech); KW-2871 (Gangioside GD3, Kyowa); MORAb-009 (GP-9, Morphotech); CDX-1307 (MDX-1307) (hCGb, Celldex); Trastuzumab (Herceptin) (HER2, Celldex); Pertuzumab (rhuMAb) 2C4)(HER2(DI), Genentech); apolizumab (HLA-DRβ chain, PDL Pharma); AMG-479 (IGF-1R, Amgen); anti-IGF-1R R1507 (IGF1-R, Roche); CP751871 (IGF1-R, Pfizer); IMC-A12 (IGF1-R, Imclone);BIIB022 (IGF-1R, Biogen); Mik-β-1 (IL-2Rb (CD122), Hoffman / LaRoche); CNTO328 (IL6, Centocor); Anti-KIR (1-7F9) (Killer Ig-like Receptor (KIR), Novo); Hu3S193 (Lewis(y), Wyeth, Ludwig Cancer Institute); hCBE-11 (LTβR, Biogen); HuHMFG1 (MUC1, Antisoma / NCL); RAV12 (N-linked carbohydrate antigen epitope, Raven); CAL (PTH-rP, University of California); CT-011 (PD1, CureTech); MDX-1106 (ono-4538) (PD1, Medarex / Ono); Mab CT-011 (PD1, Curetech); IMC-3G3 (PDGFRa, Imclone); bavituximab (phosphatidylserine, Peregrine); huJ591 (PSMA, Cornell Research Foundation); muJ591 (PSMA, Cornell Research Foundation); GC1008 (TGFb (pan-)inhibitor (IgG4), Genzyme); infliximab (Remicade) (TNFa, Centocor); A27.15 (transferrin receptor, Salk Institute, INSERN WO 2005 / 111082); E2.3 (transferrin receptor, Salk Institute); bevacizumab (Avastin) (VEGF, Genentech); HuMV833 (VEGF, Tsukuba Research Lab, PCT Publication No. WO / 2000 / 034337, University) ofTexas); IMC-18F1 (VEGFR1, Imclone); IMC-1121 (VEGFR2, Imclone). ;

[0312] Examples of useful bispecific antibodies include, but are not limited to, those that have one antibody against a tumor cell antigen and another against a cytotoxic triggering molecule, such as anti-FcγRI / anti-CD15, anti-p185. HER2 / FcγRIII (CD16), anti-CD3 / anti-malignant B cell (1D10), anti-CD3 / anti-p185 HER2Anti-CD3 / anti-p97, anti-CD3 / anti-renal cell carcinoma, anti-CD3 / anti-OVCAR-3, anti-CD3 / L-D1 (anti-colon cancer), anti-CD3 / anti-melanocyte-stimulating hormone analog, anti-EGF receptor / anti-CD3, anti-CD3 / anti-CAMA1, anti-CD3 / anti-CD19, anti-CD3 / MoV18, anti-neuronal cell adhesion molecule (NCAM) / anti-CD3, anti-folate-binding protein (FBP) / anti-CD3, anti-pan-cancer-associated antigen (AMOC-31) / anti-CD3; with Bispecific antibodies, which contain one antibody that specifically binds to tumor antigens and another antibody that binds to toxins, such as anti-saponin / anti-Id-1, anti-CD22 / anti-saponin, anti-CD7 / anti-saponin, anti-CD38 / anti-saponin, anti-CEA / anti-ricin A chain, anti-interferon α (IFN-α) / anti-hybridoma idiotype, and anti-CEA / anti-vinca alkaloids; bispecific antibodies used as prodrugs for converting enzyme activation, such as anti-CD30 / anti-alkaline phosphatase (which converts mitomycin phosphate). Bispecific antibodies that can be used as fibrinolytic agents include: prodrugs catalyzed for conversion to mitomycinol (phosphate); antifibrinogen / antitissue plasminogen activator (tPA), antifibrinogen / antiurokinase plasminogen activator (uPA); bispecific antibodies targeting cell surface receptors such as anti-low-density lipoprotein (LDL) / anti-Fc receptors (e.g., FcγRI, FcγRII, or FcγRIII); bispecific antibodies for treating infectious diseases such as anti-CD3 / anti-herpes simplex virus (HSV), anti-T cell receptor: CD3 complex / anti-influenza, anti-FcγR / anti-HIV; and bispecific antibodies for in vitro or in vivo tumor detection such as anti-CEA / anti-EOTUBE, anti-CEA / anti-DPTA, and anti-p185. HER2 / Anti-half-antibody; bispecific antibodies as vaccine adjuvants (see Fanger, MW et al., Crit Rev Immunol. 1992; 12(34):101-24, which is incorporated herein by reference); and bispecific antibodies as diagnostic tools, such as anti-rabbit IgG / antiferritin, anti-hormone peroxidase (HRP) / antihormone, anti-growth accretastatin / antisubstance P, anti-HRP / anti-FITC, anti-CEA / anti-β-galactosidase (see Nolan, O et R. O'Kennedy, Biochim Biophys Acta. 1990 Aug. 1; 1040(1):1-11, which is incorporated herein by reference). Examples of trispecific antibodies include anti-CD3 / anti-CD4 / anti-CD37, anti-CD3 / anti-CD5 / anti-CD37, and anti-CD3 / anti-CD8 / anti-CD37.

[0313] Modified amino acids and non-natural amino acids In some embodiments, the amino acid residues have a structure shown in one of the following general formulas: .

[0314] Those skilled in the art will understand that antibodies are typically composed of L-amino acids. However, for non-natural and modified amino acids, the methods and compositions of this invention provide practitioners with the ability to use L-, D-, or racemic non-natural amino acids at site-specific locations. In some embodiments, the non-natural or modified amino acids described in this invention include both the D-form and racemic forms of natural amino acids.

[0315] In the above general formula, the wavy line represents the bond connecting to the remainder of the antibody polypeptide chain. These non-natural or modified amino acids can be incorporated into the polypeptide chain in the same way as natural or modified amino acids are incorporated into the same polypeptide chain. In some embodiments, the non-natural or modified amino acids are incorporated into the polypeptide chain via amide bonds as shown in the general formula.

[0316] In the above general formula, R refers to any unrestricted functional group, as long as the amino acid residue is not identical to a native amino acid residue. In some embodiments, R can be a hydrophobic group, hydrophilic group, polar group, acidic group, basic group, chelating group, active group (reactive group), therapeutic group portion, or labeling group portion. In some embodiments, R is selected from the group consisting of: R 1a NR 2a R 3a R 1a C(=O)R 2a R 1a C(=O)OR 2a R 1a N3, and R 1a C(≡CH). In these implementation schemes, R 1a Choose from the group consisting of: bond, alkylene, heteroalkylene, arylene, and heteroarylene. R 2a and R 3a Each is independently selected from the following groups: H, alkyl, and heteroalkyl.

[0317] In some embodiments, the non-naturally encoded amino acid or modified amino acid includes a side-chain functional group that reacts effectively and selectively with functional groups (including but not limited to azido, ketone, aldehyde, and aminooxy groups) not present in 20 common amino acids to form a stable conjugate. For example, an antigen-binding polypeptide containing a non-naturally encoded amino acid with an azido functional group can be reacted with a polymer (including but not limited to polyethylene glycol) or with a second polypeptide containing an alkyne moiety to form a stable conjugate, i.e., through the selective reaction of the azido group and the alkyne functional group, thereby forming a Huisgen [3+2] cycloaddition product.

[0318] Exemplary non-naturally encoded amino acids or modified amino acids suitable for use in the present invention and capable of reacting with water-soluble polymers include, but are not limited to, those that react with carbonyl, aminooxy, hydrazine, acylhydrazine, aminourea, azide, and alkyne active groups. In some embodiments, the non-naturally encoded amino acids or modified amino acids comprise a glycosidic moiety. Examples of such amino acids include... N -Acetyl-L-glucosinolate-L-serine N -Acetyl-L-galactosaminyl-L-serine N -Acetyl-L-glucosyl-L-threonine, N -acetyl-L-glucosyl-L-asparagine, and O -mannosaminyl-L-serine. Examples of this type of amino acid also include those naturally occurring between amino acids and sugars. N -or O - Examples where the linkage is replaced by a covalent bond that is not commonly found in nature (including but not limited to alkenes, oximes, thioethers, amides, etc.). Examples of such amino acids also include sugars that are not commonly found in naturally occurring proteins, such as 2-deoxyglucose and 2-deoxygalactose.

[0319] Many of the non-naturally encoded amino acids or modified amino acids provided in this invention are commercially available, for example, from Sigma-Aldrich (St. Louis, Mo., USA), Novabiochem (a division of EMD Biosciences, Darmstadt, Germany), or Peptech (Burlington, Mass., USA). For amino acids that are not commercially available, they may optionally be synthesized according to the methods described in this invention, or synthesized using standard methods known to those skilled in the art. For organic synthesis techniques, see, for example, *Organic Chemistry* by Fessendon and Fessendon, (1982, Second Edition, Willard Grant Press, Boston Mass.); *Advanced Organic Chemistry* by March (Third Edition, 1985, Wiley and Sons, New York); and *Advanced Organic Chemistry* by Carey and Sundberg (Third Edition, Parts A and B, 1990, Plenum Press, New York). See also U.S. Patent Application Publications US2003 / 0082575 and US2003 / 0108885, both of which are incorporated herein by reference. In addition to non-natural amino acids containing non-natural side chains, the non-natural amino acids to which this invention is applicable may optionally comprise modified main-chain structures, including but not limited to structures shown in Formulas II-1 and III-1: Z typically contains OH, NH2, SH, NH-R', or S-R'; X b and Y bThe R and R' groups can be the same or different, and typically contain S or O respectively; R and R' can be the same or different, and typically selected from the same member list of R groups of non-natural amino acids having the structure shown in Formula II as described in this invention, and H respectively. For example, the non-natural amino acids or modified amino acids of this invention may optionally include substitutions on the amino or carboxyl groups, as shown in Formulas II-I and III-I. Such non-natural amino acids or modified amino acids include, but are not limited to, α-hydroxy acids, α-thio acids, and α-aminothiocarboxylic acid esters, whose side chains include, but are not limited to, side chains corresponding to the 20 common natural or non-natural amino acids. In addition, substitutions on the α-carbon may optionally include, but are not limited to, L-, D-, or α-α-disubstituted amino acids, such as D-glutamic acid, D-alanine, D-methyl-O-tyrosine, and GABA. Other structural substitutes include cyclic amino acids, such as proline analogs and cyclic proline analogs consisting of 3, 4, 6, 7, 8, and 9 atoms, and P and γ amino acids, such as substituted β-alanine and γ-aminobutyric acid.

[0320] Many non-natural amino acids or modified amino acids are based on natural amino acids (e.g., tyrosine, glutamine, phenylalanine, etc.) and are all suitable for this invention. Tyrosine analogues include, but are not limited to, para-substituted tyrosine, ortho-substituted tyrosine, and meta-substituted tyrosine, wherein the substituted tyrosine includes, but is not limited to, ketone groups (including but not limited to acetyl groups), benzoyl groups, amino groups, hydrazine, hydroxylamine, thiol groups, carboxyl groups, isopropyl groups, methyl groups, C6-C... 20Straight-chain or branched hydrocarbons, saturated or unsaturated hydrocarbons, O-methyl groups, polyether groups, nitro groups, alkynyl groups, etc. Furthermore, multi-substituted aryl rings are also contemplated. Glutamine analogs suitable for this invention include, but are not limited to, α-hydroxy derivatives, γ-substituted derivatives, cyclic derivatives, and amide-substituted glutamine derivatives. Examples of phenylalanine analogs suitable for this invention include, but are not limited to, para-substituted phenylalanines, ortho-substituted phenylalanines, and meta-substituted phenylalanines, wherein the substituents include, but are not limited to, hydroxyl groups, methoxy groups, methyl groups, allyl groups, aldehyde groups, azide groups, iodo groups, bromo groups, ketone groups (including but not limited to acetyl groups), benzoyl groups, alkynyl groups, etc. Specific examples of non-natural amino acids applicable to this invention include, but are not limited to, p-acetyl-L-phenylalanine, O-methyl-L-tyrosine, L-3-(2-naphthyl)alanine, 3-methylphenylalanine, O-4-allyl-L-tyrosine, 4-propyl-L-tyrosine, tri-O-acetyl-GlcNAcβ-serine, L-DOPA, fluorinated phenylalanine, isopropyl-L-phenylalanine, p-azido-L-phenylalanine, p-azido-methyl-L-phenylalanine, p-acyl-L-phenylalanine, p-benzoyl-L-phenylalanine, L-phosphoserine, phosphonoserine, phosphonotyrosine, p-iodophenylalanine, p-bromophenylalanine, p-amino-L-phenylalanine, isopropyl-L-phenylalanine, and p-propynoxy-phenylalanine. Structural examples of various non-natural amino acids applicable to this invention are provided, for example, see WO 2002 / 085923, entitled “In vivo incorporation of unnatural amino acids”. See also Kiick et al., (2002) Incorporation of azides into recombinant proteins for chemoselective modification by the Staudingerligation, PNAS 99:19-24, which also includes other methionine analogs.

[0321] Many of the non-natural or modified amino acids suitable for use in this invention are commercially available, for example from Sigma (USA) or Aldrich (Milwaukee, Wisconsin, USA). Those compounds that are not commercially available may optionally be synthesized as provided in this invention or in various publications, or using standard methods known to those skilled in the art. For information on organic synthesis techniques, see, for example, Fessendon and Fessendon, “Organic Chemistry” (1982, Second Edition, Willard Grant Press, Boston Mass.); March, “Advanced Organic Chemistry” (Third Edition, 1985, Wiley and Sons, New York); and Carey and Sundberg, “Advanced Organic Chemistry” (Third Edition, Parts A and B, 1990, Plenum Press, New York). Other publications describing the synthesis of unnatural amino acids include, for example, WO 2002 / 085923, entitled “Invivo incorporation of Unnatural Amino Acids”; Matsoukas et al., (1995) J. Med. Chem., 38, 4660-4669; King, FE & Kidd, DAA (1949) A New Synthesis of Glutamine and of γ-Dipeptides of Glutamic Acid from Phthylated Intermediates. J. Chem. Soc., 3315-3319; Friedman, OM & Chatterrji, R. (1959) Synthesis of Derivatives of Glutamine as Model Substrates for Anti-Tumor Agents. J. Am. Chem. Soc. 81, 3750-3752; Craig, JC et al.(1988) Absolute Configuration of the Enantiomers of 7-Chloro-4 [[4-(diethylamino)-1-methylbutyl]amino]quinoline (Chloroquine). J. Org. Chem. 53,1167-1170; Azoulay,M., Vilmont, M.&Frappier, F. (1991) Glutamine analogues asPotential Antimalarials, Eur. J. Med. Chem. 26, 201-5; Koskinen,A. M. P.&Rapoport, H. (1989) Synthesis of 4-Substituted Prolines as ConformationallyConstrained Amino Acid Analogues. J. Org. Chem. 54, 1859-1866; Christie, B.D.&Rapoport, H. (1985) Synthesis of Optically Pure Pipecolates from L-Asparagine. Application to the TotalSynthesis of (+)-Apovincamine throughAmino Acid Decarbonylation and Iminium Ion Cyclization. J. Org. Chem. 1989:1859-1866; Bartonet al., (1987) Synthesis of Novel a-Amino-Acids andDerivatives Using Radical Chemistry: Synthesis of L- and D-a-Amino-AdipicAcids, L-a-aminopimelic Acidand Appropriate Unsaturated Derivatives.Tetrahedron Lett. 43:4297-4308; and Subasinghe et al.(1992) Quisqualic acid analogues: synthesis of beta-heterocyclic 2-aminopropanoic acid derivatives and their activity at a novel quisqualate-sensitized site. J. Med. Chem. 35:4602-7. See also U.S. patent applications entitled "Protein Arrays," filed December 22, 2003, Serial No. 10 / 744,899, and December 22, 2002, Serial No. 60 / 435,821.

[0322] Amino acids with carbonyl active (reactive) groups can be linked into molecules (including but not limited to PEG or other water-soluble molecules) via a variety of reactions such as nucleophilic addition or aldol condensation.

[0323] Examples of carbonyl-containing amino acids are shown below: Where n3 is 0-10; R 1b It is an alkyl, aryl, substituted alkyl, or substituted aryl; R 2b It is H, alkyl, aryl, substituted alkyl, or substituted aryl; and R 3b It is H, amino acid, polypeptide, or amino-terminal modifying group, and R 4b It is an H, amino acid, polypeptide, or carboxyl-terminal modifying group. In some embodiments, n3 is 1, R 1b It is phenyl, and R 2b It is a simple alkyl group (i.e., methyl, ethyl, or propyl), and the ketone group is located at the para position relative to the alkyl side chain. In some embodiments, n3 is 1, R 1b It is phenyl, and R 2b It is a simple alkyl group (i.e., methyl, ethyl, or propyl), and the ketone group is located in the meta position relative to the alkyl side chain.

[0324] In some embodiments, non-naturally encoded amino acids or modified amino acids with adjacent hydroxyl and amino groups can be introduced into the peptide as "masked" aldehyde functional groups. For example, 5-hydroxylysine has a hydroxyl group adjacent to ε-amino. The reaction conditions for generating the aldehyde typically involve adding an excess of sodium periodate under mild conditions to prevent oxidation at other sites within the peptide. The pH of the oxidation reaction is typically about 7.0. A typical reaction involves adding about 1.5 molar excess sodium periodate to a buffer solution of the peptide and then incubating in the dark for about 10 minutes. See, for example, U.S. Patent No. US6,423,685, which is incorporated herein by reference.

[0325] Under mild aqueous conditions, the carbonyl functional group can selectively react with reagents containing hydrazine, acylhydrazine, hydroxylamine, or aminourea to form corresponding hydrazone, oxime, or saccharin bonds, all of which are stable under physiological conditions. See, for example, Jencks, WP, J. Am. Chem. Soc. 81, 475-481 (1959); Shao, J. and Tam, JP, J. Am. Chem. Soc. 117:3893-3899 (1995). Furthermore, the unique reactivity of the carbonyl group allows for selective modification in the presence of other amino acid side chains. See, for example, Cornish, VW, et al., J.Am. Chem. Soc. 118:8150-8151 (1996); Geoghegan, KF & Stroh, JG, Bioconjug. Chem. 3:138-146 (1992); Mahal, LK, et al., Science 276:1125-1128 (1997).

[0326] Non-naturally encoded amino acids or modified amino acids containing nucleophilic groups (such as hydrazine, acylhydrazine, or aminourea) can react with various electrophilic groups to form couplings (including, but not limited to, reactions with PEG or other water-soluble polymers).

[0327] Examples of amino acids containing hydrazine, acylhydrazine, or aminourea can be represented as follows: Where n3 is 0-10; R 1c It is an alkyl, aryl, substituted alkyl, or substituted aryl, or R 1c Does not exist; X c Is it O, N, S, or X? c Does not exist; R 2c It is H, amino acid, polypeptide, or amino-terminal modifying group; and R3c It is an H, amino acid, polypeptide, or carboxyl terminator.

[0328] In some implementations, n3 is 4, R 1c It does not exist, and X is N. In some implementations, n3 is 2, and R... 1c It does not exist, and X does not exist. In some implementations, n3 is 1, R 1c It is a phenyl group, X is O, and the oxygen atom is located at the para position of an aliphatic group on the aromatic ring.

[0329] Amino acids containing acylhydrazides, hydrazides, and aminoureas are available from commercial sources. For example, L-glutamic acid-γ-acylhydrazides are available from Sigma Chemical (St. Louis, Mo.). Other amino acids that are not commercially available can be prepared by those skilled in the art. See, for example, U.S. Patent No. 6,281,211, which is incorporated herein by reference.

[0330] Peptides containing non-naturally encoded amino acids or modified amino acids (with acylhydrazide, hydrazine, or aminourea functional groups) can react efficiently and selectively with a variety of molecules containing aldehydes or other functional groups with similar chemical reactivity. See, for example, Shao, J. and Tam, J., J. Am. Chem. Soc. 117:3893-3899 (1995). The unique reactivity of acylhydrazide, hydrazine, and aminourea functional groups makes them significantly more reactive to aldehydes, ketones, and other electrophilic groups than to nucleophilic groups on 20 common amino acids (including, but not limited to, the hydroxyl groups of serine or threonine, or the amino groups and N-termini of lysine).

[0331] Non-naturally encoded amino acids or modified amino acids containing an aminooxy group (also known as hydroxylamine) can react with a variety of electrophilic groups to form couplings (including, but not limited to, reactions with PEG or other water-soluble polymers). Similar to hydrazine, acylhydrazine, and aminourea, the enhanced nucleophilicity of the aminooxy group allows it to react efficiently and selectively with a variety of molecules containing aldehydes or other functional groups with similar chemical reactivity. See, for example, Shao, J. and Tam, J., J. Am.Chem. Soc. 117:3893-3899 (1995); H. Hang and C. Bertozzi, Acc. Chem. Res. 34:727-736 (2001). The product of reaction with a hydrazine group is the corresponding hydrazone, while the product of reaction with an aminooxy group and a carbonyl-containing group (e.g., a ketone) is typically an oxime.

[0332] An example amino acid containing an amino group can be represented as follows: Where n3 is 0-10; R 1c It is an alkyl, aryl, substituted alkyl, or substituted aryl, or R 1c Does not exist; X c Is it O, N, S, or X? c It does not exist; m3 is 0-10; Y c It is ═C(O), or Y c Does not exist; R 2c It is H, amino acid, polypeptide, or amino-terminal modifying group; and R 3c It is an H, amino acid, polypeptide, or carboxyl-terminal modifying group. In some embodiments, n3 is 1, R 1a It is phenyl, X c It is O, m is 1, and Y c Exists. In some implementations, n3 is 2, R 1c and X c Neither exists, m3 is 0, and Y c It does not exist.

[0333] Aminooxylated amino acids can be prepared from readily available amino acid precursors (homoserine, serine, and threonine). See, for example, M. Carrasco and R. Brown, J. Org. Chem. 68: 8853-8858 (2003). Some aminooxylated amino acids, such as L-2-amino-4-(aminooxy)butyric acid, have been isolated from natural sources (Rosenthal, G. et al., Life Sci. 60: 1635-1641 (1997)). Other aminooxylated amino acids can be prepared by those skilled in the art.

[0334] The unique reactivity of azide and alkyne functional groups makes them extremely useful for the selective modification of peptides and other biomolecules. Organic azides, particularly aliphatic azides and alkynes, are generally stable under common reaction chemistry conditions. In particular, the azide and alkyne functional groups are inert to the side chains (i.e., R groups) of the 20 most common amino acids found in naturally occurring peptides. However, when they come into close proximity, the "spring-loaded" nature of the azide and alkyne groups emerges, allowing them to react selectively and efficiently via the Huisgen [3+2] cycloaddition reaction to generate the corresponding triazoles. See, for example, Chin J., et al., Science 301:964-7 (2003); Wang, Q., et al., J. Am. Chem. Soc.125, 3192-3193 (2003); Chin, JW, et al., J. Am. Chem. Soc. 124:9026-9027 (2002).

[0335] Because Huisgen's cycloaddition reactions involve selective cycloaddition (see, for example, Padwa, A., in COMPREHENSIVE ORGANIC SYNTHESIS, Vol. 4, (ed. Trost, BM, 1991), pp. 1069-1109; Huisgen, R., in 1,3-DIPOLAR CYCLOADDITION CHEMISTRY, (ed. Padwa, A., 1984), pp. 1-176) rather than nucleophilic substitution, the incorporation of non-naturally encoded or modified amino acids with azide and alkyne side chains results in selective modification of the resulting polypeptide at the positions of the non-naturally encoded or modified amino acids. Cycloaddition reactions of azide- or alkyne-containing antibodies can be carried out at room temperature, in aqueous conditions, by adding Cu(II) (including, but not limited to, a catalytic amount of CuSO4) in the presence of a reducing agent to reduce the catalytic amount of Cu(II) to Cu(I) in situ. See, for example, Wang, Q., et al., J. Am. Chem. Soc. 125, 3192-3193 (2003); Tornee, CW, et al., J. Org. Chem. 67:3057-3064 (2002); Rostovtsev, et al., Angew. Chem. Int. Ed. 41:2596-2599 (2002). Exemplary reducing agents include, but are not limited to, ascorbic acid salts, metallic copper, quinine, hydroquinone, vitamin K, glutathione, cysteine, and Fe. 2+Co 2+ and the applied electric potential.

[0336] In some cases, when a Huisgen [3+2] cycloaddition reaction between an azide and an alkyne is required, the antigen-binding polypeptide comprises a non-naturally encoded amino acid or a modified amino acid containing an alkyne moiety, and the water-soluble polymer to be linked to the amino acid contains an azide moiety. Alternatively, the reverse reaction can also be performed (i.e., the azide moiety on the amino acid reacts with the alkyne moiety on the water-soluble polymer).

[0337] The azide functional group can also selectively react with water-soluble polymers containing aryl esters and be partially and appropriately functionalized with arylphosphine groups to form amide bonds. The arylphosphine group reduces the azide in situ, and the resulting amine then reacts efficiently with the proximal ester bond to form the corresponding amide. See, for example, E. Saxon and C. Bertozzi, Science 287, 2007-2010 (2000). The azide-containing amino acid can be an alkyl azide (including, but not limited to, 2-amino-6-azido-1-hexanoic acid) or an aryl azide (p-azido-phenylalanine).

[0338] An exemplary water-soluble polymer containing aryl ester and phosphine group moieties can be represented as follows: Where X c It can be O, N, S, or X cNo, Ph is phenyl, W is a water-soluble polymer, and R can be H, alkyl, aryl, substituted alkyl, and substituted aryl groups. Exemplary R groups include, but are not limited to, –CH2, –C(CH3)3, –OR′, –NR′R′′, –SR′, –halogen, –C(O)R′, –CONR′R′′, –S(O)2R′, –S(O)2NR′R′′, –CN, and –NO2. R′, R′′, R′′′, and R′′′′ each independently refer to H, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl (including, but not limited to, aryl substituted with 1-3 halogens), substituted or unsubstituted alkyl, alkoxy or thioalkoxy, or aralkyl groups. When the compounds of the present invention include more than one R group, for example, when more than one of these groups is present, the R groups are each independently selected as individual R′, R′′, R′′′, and R′′′′ groups. When R' and R'' are attached to the same nitrogen atom, they can combine with the nitrogen atom to form a ring consisting of 5, 6, or 7 atoms. For example, -NR′R′′ is intended to include, but is not limited to, 1-pyrrolidinyl and 4-morpholinyl. Based on the above discussion of substituents, those skilled in the art will understand that the term "alkyl" is intended to include groups comprising a carbon atom attached to a group other than a hydrogen group, such as haloalkyl (including but not limited to –CF3 and –CH2CF3) and acyl (including but not limited to –C(O)CH3, –C(O)CF3, –C(O)CH2OCH3, and the like).

[0339] The azide functional group can also selectively react with water-soluble polymers containing thioesters and be appropriately functionalized with an arylphosphine group to generate an amide bond. The arylphosphine group reduces the azide in situ, and the resulting amine reacts effectively with the thioester bond to generate the corresponding amide. An exemplary water-soluble polymer containing both a thioester and a phosphine group moiety can be represented as follows: Where n2 is 1-10; X c It can be O, N, or S, or X c It does not exist; Ph is phenyl; and W is a water-soluble polymer.

[0340] An example of an alkyne-containing amino acid can be represented as follows: Where n3 is 0-10; R 1c It is an alkyl, aryl, substituted alkyl, or substituted aryl, or R 1c Does not exist; X c Is it O, N, S, or X? c It does not exist; m3 is 0-10, R 2c It is H, amino acid, polypeptide, or amino-terminal modifying group, and R3c It is an H, amino acid, polypeptide, or carboxyl-terminal modifying group. In some embodiments, n3 is 1, R 1c It is phenyl, X c It does not exist; m3 is 0, and the acetylene group is located at the para position relative to the alkyl side chain. In some embodiments, n3 is 1, R 1c It is phenyl, X c It is O, m3 is 1, and the propargyloxy group is located at the para position relative to the alkyl side chain (i.e., O-propargyl-tyrosine). In some embodiments, n3 is 1, R 1c and X c Neither exists, and m3 is 0 (i.e., propargyl glycine).

[0341] Alkyne-containing amino acids are commercially available. For example, propargyl glycine is available from Peptech (Burlington, Mass.). Alternatively, alkyne-containing amino acids can be prepared according to standard methods. For example, p-propargyloxyphenylalanine can be synthesized, as described in Deiters, A., et al., J. Am. Chem. Soc. 125: 11782-11783 (2003), and 4-alkynyl-L-phenylalanine can be synthesized, as described in Kayser, B., et al., Tetrahedron 53(7): 2475-2484 (1997). Other alkyne-containing amino acids can also be prepared by those skilled in the art.

[0342] An example of an amino acid containing an azide can be represented as follows: Where n3 is 0-10; R 1c It is an alkyl, aryl, substituted alkyl, or substituted aryl, or R 1c Does not exist; X c Is it O, N, S, or X? c It does not exist; m3 is 0-10; R 2c It is H, amino acid, polypeptide, or amino-terminal modifying group; and R 3c It is an H, amino acid, polypeptide, or carboxyl-terminal modifying group. In some embodiments, n3 is 1, R 1c It is phenyl, X c It does not exist; m3 is 0, and the azide group is located at the para position of the alkyl side chain. In some embodiments, n3 is 0-4, R 1c and X c Neither exists, and m3 = 0. In some implementations, n3 is 1, and R... 1c It is phenyl, X cIt is O, m3 is 2, and the para-azidoethoxy group is located at the para position relative to the alkyl side chain.

[0343] Azide-containing amino acids are available from commercial sources. For example, 4-azidophenylalanine is available from Chem-Impex International (Wood Dale, IL). For amino acids containing azides that are not commercially available, the azide group can be prepared relatively readily using standard methods known to those skilled in the art, including but not limited to substitution of a suitable leaving group (including but not limited to halides, methanesulfonates, toluenesulfonates) or by opening a suitably protected lactone. See, for example, Advanced Organic Chemistry by March (Third Edition, 1985, Wiley and Sons, New York).

[0344] The unique reactivity of β-substituted aminothiol functional groups makes them well-suited for the selective modification of aldehyde-containing peptides and other biomolecules by forming thiazolidinyl esters. See, for example, J. Shao and J. Tam, J. Am. Chem. Soc. 1995, 117 (14) 3893-3899. In some embodiments, β-substituted aminothiol amino acids can be incorporated into antibodies and then reacted with water-soluble polymers containing aldehyde functional groups. In some embodiments, water-soluble polymers, drug conjugates, or other payloads can be conjugated to antibody peptides containing β-substituted aminothiol amino acids by forming thiazolidinyl esters.

[0345] Specific examples of useful modified amino acids include, but are not limited to, p-acetyl-L-phenylalanine, O-methyl-L-tyrosine, L-3-(2-naphthyl)alanine, 3-methyl-phenylalanine, O-4-allyl-L-tyrosine, 4-propyl-L-tyrosine, tri-O-acetyl-GlcNAcβ-serine, L-DOPA, fluorinated phenylalanine, isopropyl-L-phenylalanine, p-azido-methyl-L-phenylalanine, p-azido-L-phenylalanine, p-acyl-L-phenylalanine, p-benzoyl-L-phenylalanine, L-phosphoserine, phosphonoserine, phosphonotyrosine, p-iodo-phenylalanine, p-bromophenylalanine, p-amino-L-phenylalanine, isopropyl-L-phenylalanine, and p-propynyloxy-phenylalanine. Other useful examples include... N -Acetyl-L-glucosyl-L-serine N -Acetyl-L-aminogalactosyl-L-serine N -Acetyl-L-glucosyl-L-threonine, N-acetyl-L-glucosyl-L-asparagine, and O -Aminomannosyl-L-serine.

[0346] In a particular embodiment, the modified amino acid is selected from p-acetyl-phenylalanine, p-ethynyl-phenylalanine, p-propynoxyphenylalanine, p-azido-methyl-phenylalanine, and p-azido-phenylalanine. Particularly useful modified amino acids are p-azido-phenylalanine and p-azido-methyl-phenylalanine (pAMF). These amino acid residues are known to those skilled in the art to facilitate, for example, Huisgen [3+2] cycloaddition reactions (so-called "click" chemistry) with compounds containing an alkynyl group. This reaction allows those skilled in the art to readily and rapidly conjugate antibodies at site-specific locations of the modified amino acid.

[0347] In some embodiments, the first active group is an alkynyl group moiety (including, but not limited to, the alkynyl group moiety in the non-natural amino acid p-propynyloxyphenylalanine, wherein the propynyl group is sometimes also referred to as the acetylene group moiety), and the second active group is an azido group moiety, which may be used with [3+2] cycloaddition chemistry. In some embodiments, the first active group is an azido group moiety (including, but not limited to, the azido group moiety in the non-natural amino acid p-azido-L-phenylalanine or p-azidomethyl-phenylalanine), and the second active group is an alkynyl group moiety.

[0348] Another useful amino acid is p-acetyl-L-phenylalanine (pAcF), which is known to those skilled in the art to promote oxime linkage. In some embodiments, the first active group is the acetyl group moiety (including, but not limited to, the acetyl group moiety in the non-natural amino acid p-acetyl-L-phenylalanine), and the second active group is the aminooxy group moiety.

[0349] In the above general formula, each L represents a divalent linker. The divalent linker can be any divalent linker known to those skilled in the art. Typically, the divalent linker is capable of forming a covalent bond with the functional group portion R and a homologous active group (e.g., α-carbon) of a non-natural amino acid or a modified amino acid. Useful divalent linkers are alkylene, substituted alkylene, heteroalkylene, substituted heteroalkylene, aryl, substituted aryl, heteroaryl, and substituted heteroaryl. In some embodiments, L is C. 1-10 Alkylene or C 1-10 Heteroalkyl groups.

[0350] The non-natural amino acids or modified amino acids used in the methods and compositions of the present invention have at least one of the following four properties: (1) at least one functional group on the side chain of the non-natural amino acid or modified amino acid has at least one characteristic and / or activity and / or reactivity orthogonal to the chemical reactivity of 20 common genetically encoded amino acids (i.e., alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine), or at least orthogonal to the chemical reactivity of naturally occurring amino acids present in polypeptides comprising non-natural amino acids or modified amino acids; (2) the introduced non-natural amino acids or modified amino acids have at least one characteristic and / or activity and / or reactivity orthogonal to the chemical reactivity of naturally occurring amino acids present in polypeptides comprising non-natural amino acids or modified amino acids; The modified amino acids are substantially chemically inert to the 20 most common genetically encoded amino acids; (3) the non-natural or modified amino acids can be stably incorporated into the polypeptide, preferably polypeptides with stability comparable to naturally occurring amino acids, or, under typical physiological conditions, more preferably, such incorporation can be carried out via an in vivo system; and (4) the non-natural or modified amino acids include oxime functional groups or functional groups that can be converted to oxime groups by reaction with a reagent, preferably under conditions that do not destroy the biological properties of the polypeptide including the non-natural or modified amino acids (unless such destruction of biological properties is for the purpose of modification / conversion), or said conversion can be carried out under aqueous conditions at a pH of about 4 to about 8, or said reactive site on the non-natural amino acid is an electrophilic site. Any number of non-natural or modified amino acids can be introduced into the polypeptide. The non-natural or modified amino acids may also include protected or masked oximes or protected or masked groups that can be converted to oxime groups after the protected group is deprotected or the masked group is demasked. Non-natural amino acids or modified amino acids may also include protected or masked carbonyl or dicarbonyl groups, which can be converted into carbonyl or dicarbonyl groups after the protected group is deprotected or the masked group is demasked, thereby reacting with hydroxylamine or aminooxy groups to form oxime groups.

[0351] In further embodiments, non-natural or modified amino acids that can be used in the methods and compositions of the present invention include, but are not limited to, amino acids containing photoactivated crosslinking agents, spin-labeled amino acids, fluorescent amino acids, metal-bound amino acids, metal-containing amino acids, radioactive amino acids, amino acids with novel functional groups, amino acids that covalently or non-covalently interact with other molecules, photo-masking and / or photoisomerizable amino acids, amino acids containing biotin or biotin analogs, glycosylated amino acids (e.g., sugar-substituted serine), amino acids modified with other carbohydrates, ketone-containing amino acids, aldehyde-containing amino acids, amino acids containing polyethylene glycol or other polyethers, heavy atom-substituted amino acids, chemically cleavable and / or photocleavable amino acids, amino acids with long side chains compared to natural amino acids (including but not limited to polyethers or long-chain hydrocarbons, including but not limited to more than about 5 or more than about 10 carbon atoms), carbon-linked sugar-containing amino acids, redox-active amino acids, amino acids containing aminothioic acids, and amino acids containing one or more toxic moieties.

[0352] In some implementations, non-natural or modified amino acids contain a sugar group moiety. Examples of such amino acids include... N -Acetyl-L-glucosyl-L-serine N -Acetyl-L-aminogalactosyl-L-serine N -Acetyl-L-glucosyl-L-threonine, N -acetyl-L-glucosyl-L-asparagine, and O -Aminomannosyl-L-serine. Examples of this type of amino acid also include those naturally occurring between amino acids and sugars. N -or O - Examples where the linkage is replaced by a covalent bond that is not commonly found in nature (including but not limited to alkenes, oximes, thioethers, amides, etc.). Examples of such amino acids also include sugars that are not commonly found in naturally occurring proteins, such as 2-deoxyglucose and 2-deoxygalactose.

[0353] Incorporating the chemical group portion of an antibody by incorporating non-natural or modified amino acids offers numerous advantages and manipulability for peptides. For example, the unique reactivity of carbonyl or dicarbonyl functional groups (including ketone or aldehyde functional groups) allows for selective modification of antibodies in vivo and in vitro using any of a variety of hydrazine- or hydroxylamine-containing reagents. For instance, heavy-atom non-natural or modified amino acids can be used for phase-determined X-ray structural data. Site-specific introduction of heavy atoms using non-natural or modified amino acids also provides selectivity and flexibility in selecting the position of heavy atoms. For example, photoreactive non-natural or modified amino acids (including, but not limited to, amino acids with benzophenone and aryl azide (including, but not limited to, phenyl azide) side chains) allow for efficient photocrosslinking of peptides in vivo and in vitro. Examples of photoreactive non-natural or modified amino acids include, but are not limited to, p-azido-phenylalanine and p-benzoyl-phenylalanine. Antibodies containing photoreactive non-natural or modified amino acids can then be arbitrarily crosslinked by providing time control through the excitation of the photoreactive group. In non-limiting examples, the methyl groups of non-natural or modified amino acids may be replaced by isotopic labels (including, but not limited to, methyl groups) as probes of local structure and dynamics, including, but not limited to, using nuclear magnetic resonance and vibrational spectroscopy.

[0354] Amino acids possessing electrophilic (reactive) groups allow for a variety of chemical reactions (including, but not limited to, nucleophilic addition reactions) to link molecules. Such electrophilic (reactive) groups include carbonyl or dicarbonyl groups (including ketone or aldehyde groups), carbonyl-like or dicarbonyl-like groups (which are similar in reactivity and structure to carbonyl or dicarbonyl groups), masked carbonyl or masked dicarbonyl groups (which can be readily converted to carbonyl or dicarbonyl groups), or protected carbonyl or protected dicarbonyl groups (which exhibit reactivity similar to carbonyl or dicarbonyl groups after deprotection). Such amino acids include those having the structure shown in formula (I-1): Wherein: A is optional, and when present, is a lower alkylene, substituted lower alkylene, lower cycloalkylene, substituted lower cycloalkylene, lower alkenyl, substituted lower alkenylene, alynylene, lower heteroalkylene, substituted heteroalkylene, lower heterocycloalkylene, substituted lower heterocycloalkylene, arylene, substituted arylene, heteroarylene, substituted heteroarylene, alkylene arylene, substituted alkylene arylene, arylalkylene, or substituted arylalkylene; B is optional, and when present, is a linker selected from the group consisting of: lower alkylene, substituted lower alkylene, lower alkenyl, substituted lower alkenylene, lower heteroalkylene, substituted lower heteroalkylene, –O–, –O–(alkylene or substituted alkylene)–, –S–, –S–(alkylene or substituted alkylene)–, –S(O) k – (where k is 1, 2, or 3), –S(O) k (alkylene or substituted alkylene)–, –C(O)–, –NS(O)2–, –OS(O)2–, –C(O)–(alkylene or substituted alkylene)–, –C(S)–, –C(S)–(alkylene or substituted alkylene)–, –N(R′)–, –NR′–(alkylene or substituted alkylene)–, –C(O)N(R′)–, –CON(R′)–(alkylene or substituted alkylene)–, –CSN(R′)–, –CSN(R′)–(alkylene or substituted alkylene)–, –N(R′)CO–(alkylene or substituted alkylene)–, –N(R′)C(O)O–, –S(O) k N(R′)–, –N(R′)C(O)N(R′)–, –N(R′)C(S)N(R′)–, –N(R′)S(O) k N(R′)–, –N(R′)–N═, ―C(R′)═N–, ―C(R′)═N–N(R′)–, –C(R′)═N–N═, –C(R′)2–N═N–, and –C(R′)2–N(R′)–N(R′)–, wherein each R′ is independently H, an alkyl group, or a substituted alkyl group; J is R is H, alkyl, substituted alkyl, cycloalkyl, or substituted cycloalkyl; each R'' is independently H, alkyl, substituted alkyl, or protecting group, or when more than one R'' group is present, the two R'' optionally form a heterocyclic alkyl group; R 1d It is H, amino protecting group, resin, amino acid, polypeptide, or polynucleotide; R 2d It is an OH group, an ester protecting group, a resin, an amino acid, a polypeptide, or a polynucleotide; R 3d and R 4d Each is independently H, halogen, lower alkyl, or substituted lower alkyl, or R. 3dand R 4d Or two Rs 3d The groups optionally form cycloalkyl or heterocycloalkyl groups; or the -ABJR groups together form bicyclic or tricyclic cycloalkyl or heterocycloalkyl groups, wherein the bicyclic or tricyclic cycloalkyl or heterocycloalkyl groups comprise at least one carbonyl group, including a dicarbonyl group, a protected carbonyl group, including a protected dicarbonyl group, or a masked carbonyl group, including a masked dicarbonyl group; or the -JR groups together form monocyclic or bicyclic cycloalkyl or heterocycloalkyl groups, wherein the monocyclic or bicyclic cycloalkyl or heterocycloalkyl groups comprise at least one carbonyl group, including a dicarbonyl group, a protected carbonyl group, including a protected dicarbonyl group, or a masked carbonyl group, including a masked dicarbonyl group; provided that A is phenylene and each R 3d When A is H, then B exists; when A is –(CH2)4– and each R… 3d When A and B are both H, then B is not –NHC(O)(CH2CH2)–; and when A and B are both absent and each R 3d When R is H, then R is not methyl. Such non-natural or modified amino acids may be in the form of salts, or may be incorporated into non-natural or modified amino acid polypeptides, polymers, polysaccharides, or polynucleotides and optionally undergo post-translational modification.

[0355] In some embodiments, the compound shown in formula (I-1) is stable in aqueous solution under weakly acidic conditions for at least one month. In some embodiments, the compound shown in formula (I-1) is stable under weakly acidic conditions for at least two weeks. In some embodiments, the compound shown in formula (I-1) is stable under weakly acidic conditions for at least five days. In some embodiments, such acidic conditions are a pH value of 2 to 8.

[0356] In some embodiments of the compound shown in formula (I-1), B is a lower alkylene, a substituted lower alkylene, –O– (alkylene or substituted alkylene)–, –C(R′)═N–N(R′)–, ―N(R′)CO–, –C(O)-, –C(R′)═N–, –C(O)– (alkylene or substituted alkylene)–, –CON(R′)– (alkylene or substituted alkylene)–, –S (alkylene or substituted alkylene)–, –S(O) (alkylene or substituted alkylene)–, or ―S(O)2 (alkylene or substituted alkylene)–. In some embodiments of the compound shown in formula (I-1), B is –O(CH2)–, –CH═N–, –CH═N–NH–, –NHCH2–, ―NHCO–, –C(O)–, –C(O)–(CH2)–, ―CONH–(CH2)–, –SCH2–, –S(═O)CH2–, or –S(O)2CH2–. In some embodiments of the compound shown in formula (I-1), R is C 1–6Alkyl or cycloalkyl. In some embodiments of the compounds shown in formula (I-1), R is –CH3, –CH(CH3)2, or cyclopropyl. In some embodiments of the compounds shown in formula (I-1), R 1d It consists of H, tert-butoxycarbonyl (Boc), and 9-fluorenylmethoxycarbonyl (Fmoc). N - Acetyl, tetrafluoroacetyl (TFA), or benzyloxycarbonyl (Cbz). In some embodiments of the compound shown in formula (I-1), R 1d It is a resin, amino acid, polypeptide, or polynucleotide. In some embodiments of the compound shown in formula (I-1), R 2d It is OH, O-methyl, O-ethyl, or O-tert-butyl. In some embodiments of the compound shown in formula (I-1), R 2d It is a resin, amino acid, polypeptide, or polynucleotide. In some embodiments of the compound shown in formula (I-1), R 2d It is a polynucleotide. In some embodiments of the compound shown in formula (I-1), R 2d It is ribonucleic acid (RNA). In some embodiments of the compound shown in formula (I-1), R 2d It is tRNA. In some embodiments of the compound shown in formula (I-1), the tRNA specifically recognizes a selector codon. In some embodiments of the compound shown in formula (I-1), the selector codon is selected from the group consisting of: amber codon, ochre codon, opal codon, unique codon, rare codon, non-natural codon, pentabase codon, and tetrabase codon. In some embodiments of the compound shown in formula (I-1), R 2d It is a repressive tRNA.

[0357] In some embodiments of the compound shown in formula (I-2), Choose from the group consisting of: (i) A is a substituted lower alkylene, C4-arylene, substituted arylene, heteroarylene, substituted heteroarylene, alkylene, substituted alkylene, arylene, or substituted arylene; B is optional, and when present, is a divalent linker chosen from the group consisting of: lower alkylene, substituted lower alkylene, lower alkenyl, substituted lower alkenyl, –O–, –O– (alkylene or substituted alkylene)–, –S–, –S(O)–, –S(O)2–, –NS(O)2–, –OS(O)2–, –C(O)–, –C(O)– (alkylene or substituted alkylene)–, –C(S)–, –N(R′)–, –C(O)N(R′)–, –CON(R′) –(alkylene or substituted alkylene)–, –CSN(R′)–, ―N(R′)CO–(alkylene or substituted alkylene)–, –N(R′)C(O)O–, –N(R′)C(S)–, –S(O)N(R′), ―S(O)2N(R′), –N(R′)C(O)N(R′)–, ―N(R′)C(S)N(R′)–, –N(R′)S(O)N(R′)–, ―N(R′)S(O)2N(R′)–, –N(R′)–N═, –C(R′)═N–N(R′)–, ―C(R′)═N–N═, –C(R′)2–N═N–, and ―C(R′)2–N(R′)–N(R′)–; (ii) A is optional, and when present, is a substituted lower alkylene, C4-arylene, substituted arylene, heteroarylene, substituted heteroarylene, alkylene, substituted alkylene, arylene, or substituted arylene; B is a divalent linker selected from the group consisting of: lower alkylene, substituted lower alkylene, lower alkenyl, substituted lower alkenyl, –O–, –O– (alkylene or substituted alkylene)–, –S-, –S(O)–, –S(O)2–, –NS(O)2–, –OS(O)2–, –C(O)–, –C(O)– (alkylene or substituted alkylene)–, –C(S)–, –N(R′)–, –C(O)N(R′)–, –CON(R′) –(alkylene or substituted alkylene)–, –CSN(R′)–, –N(R′)CO–(alkylene or substituted alkylene)–, –N(R′)C(O)O–, –N(R′)C(S)–, –S(O)N(R′), –S(O)2N(R′), –N(R′)C(O)N(R′)–, –N(R′)C(S)N(R′)–, –N(R′)S(O)N(R′)–, –N(R′)S(O)2N(R′)–, –N(R′)–N═, –C(R′)═N–N(R′)–, –C(R′)═N–N═, –C(R′)2–N═N–, and –C(R′)2–N(R′)–N(R′)–; (iii) A is a lower alkylene;B is optional, and when present, is a divalent linker selected from the group consisting of: lower alkylene, substituted lower alkylene, lower alkenyl, substituted lower alkenyl, –O–, –O– (alkylene or substituted alkylene)–, –S–, –S(O)–, –S(O)2–, –NS(O)2–, –OS(O)2–, –C(O)–, –C(O)– (alkylene or substituted alkylene)–, –C(S)–, –N(R′)–, –C(O)N(R′)–, –CSN(R′)–, –CON(R′)– (alkylene or substituted alkylene)– Alkyl)–, –N(R′)C(O)O–, –N(R′)C(S)–, –S(O)N(R′), –S(O)2N(R′), –N(R′)C(O)N(R′)–, –N(R′)C(S)N(R′)–, –N(R′)S(O)N(R′)–, –N(R′)S(O)2N(R′)–, –N(R′)–N═, –C(R′)═N–N(R′)–, –C(R′)═N–N═, –C(R′)2–N═N–, and ―C(R′)2–N(R′)–N(R′)–; and (iv) A is a phenylene; B is a divalent linker selected from the group consisting of: lower alkylene, substituted lower alkylene, lower alkenyl, substituted lower alkenyl, –O–, –O– (alkylene or substituted alkylene)–, –S–, –S(O)–, –S(O)2–, –NS(O)2–, –OS(O)2–, –C(O)–, –C(O)– (alkylene or substituted alkylene)–, –C(S)–, –N(R′)–, –C(O)N(R′)-, –CON(R′)– (alkylene or substituted alkylene)–, –CSN(R′)–, –N(R′)CO –(alkylene or substituted alkylene)–, –N(R′)C(O)O–, –N(R′)C(S)–, –S(O)N(R′), –S(O)2N(R′), –N(R′)C(O)N(R′)–, –N(R′)C(S)N(R′)–, –N(R′)S(O)N(R′)–, –N(R′)S(O)2N(R′)–, –N(R′)–N═, –C(R′)N–N(R′)–, –C(R′)═N–N═, –C(R′)2–N═N–, and –C(R′)2–N(R′)–N(R′)–; J is; Each R' is independently H, an alkyl group, or a substituted alkyl group; R 1d It is optional, and when present, it is H, an amino protecting group, a resin, an amino acid, a polypeptide, or a polynucleotide; R 2d It is optional, and when present, it is an OH group, an ester protecting group, a resin, an amino acid, a polypeptide, or a polynucleotide; each R 3d and R4d Each is independently H, halogen, lower alkyl, or substituted lower alkyl; and R is H, alkyl, substituted alkyl, cycloalkyl, or substituted cycloalkyl.

[0358] In some embodiments, the non-natural amino acid or modified amino acid may have the structure shown in Formula XIX: Formula XIX; Or its salt, wherein: D a Yes-Ar-W 3a –or–W 1a -Y 1a -C(O)-Y 2a -W 2a –;Ar is W 1a W 2a and W 3a Each is independently a single bond or a lower alkylene group; each X 1b Each is independently –NH–, –O–, or –S–; each Y 1a Each is independently a single bond, –NH–, or –O–; each Y 2a Each is independently a single bond, –NH–, –O–, or N-linked or C-linked pyridine alkyl group; and Z 1 Z 2 and Z 3 One of them is –N–, and Z 1 Z 2 and Z 3 The other two are independently –CH–. In some embodiments, the non-natural amino acid or modified amino acid has the structure shown in Formula XIXa: Formula XIXa; Where D a The definition is given in the context of Formula XIX. In some embodiments, the non-natural amino acid or modified amino acid may have the structure shown in Formula XIXb: Formula XIXb; or its salt, wherein W 4a It is C1-C 10 Alkylene. In a further embodiment, W 4a It is a C1-C5 alkylene group. In one embodiment, W 4a It is a C1-C3 alkylene group. In one embodiment, W 4aIt is a C1 alkylene group. In a particular embodiment, the non-natural amino acid or modified amino acid is selected from the group consisting of: Or its salts. Such non-natural amino acids or modified amino acids may be in the form of salts, or may be incorporated into non-natural amino acids or modified amino acid peptides, polymers, polysaccharides, or polynucleotides and optionally post-translational modified.

[0359] In some embodiments, the modified amino acid has the structure shown in Formula I: Or its salt, wherein Ar is: V is a single bond, a lower alkylene group, or –W 1a –W 2a –;W 1a and W 2a One of them is absent or is a lower alkylene group, and the other is –NH–, –O–, or –S–; each X 1b Each can be independently represented as –NH–, –O–, or –S–; Z 1 Z 2 and Z 3 One of them is –CH– or –N–, Z 1 Z 2 and Z 3 The other two in are each independently –CH–; and R is a lower alkyl group. In some embodiments, when Ar is… When V is –NH–, then Z 1 Z 2 and Z 3 One of them is –N–. In some implementations, V is a single bond, –NH–, or –CH2NH–.

[0360] In some implementations, Ar is and Z 1 Z 2 Z 3 and X 1bAll have the definitions described in the context of Equation I. In some embodiments according to this paragraph, V is –W 1a –W 2a –;W 1a and W 2a One of them is absent or is –CH2–, and the other is –NH–, –O–, or –S–. In some embodiments according to this paragraph, V is a single bond, –NH–, or –CH2NH–. In some embodiments according to this paragraph, Z 1 It is N. In some embodiments according to this paragraph, Z 2 It is N. In some embodiments according to this paragraph, Z 3 It is N. In some embodiments according to this paragraph, Z 1 It is CH, Z 3 It's CH, and X 1b It is S.

[0361] In some implementations, Ar is and Z 1 Z 2 and Z 3 All have the definitions described in the context of Equation I. In some embodiments according to this paragraph, V is –W 1a –W 2a –;W 1a and W 2a One of them is absent or is –CH2–, and the other is –NH–, –O–, or –S–. In some embodiments according to this paragraph, V is a single bond, –NH–, or –CH2NH–. In some embodiments according to this paragraph, Z 1 It is N. In some embodiments according to this paragraph, Z 2 It is N. In some embodiments according to this paragraph, Z 3 It is N.

[0362] In some implementations, Ar is and Z 1 Z 3 and X 1 All have the definitions described in the context of Equation I-1. In some embodiments according to this paragraph, V is –W 1a –W 2a –;W 1a and W 2a One of them is absent or is –CH2–, and the other is –NH–, –O–, or –S–. In some embodiments according to this paragraph, V is a single bond, –NH–, or –CH2NH–. In some embodiments according to this paragraph, Z 1It is N. In some embodiments according to this paragraph, Z 3 It is N. In some embodiments according to this paragraph, Z 1 It is CH, Z 3 It's CH, and X 1b It is S.

[0363] In some embodiments, the modified amino acid has the structure shown in Formula I-1a: Ar, V, and R all have the definitions described in the context of Equation I-1.

[0364] In one embodiment, the present invention provides a compound of either formula I-1 or I-1a, wherein V is a single bond. In another embodiment, the present invention provides a compound of either formula I-1 or I-1a, wherein V is –NH–. In yet another embodiment, the present invention provides a compound of either formula I-1 or I-1a, wherein V is –CH2NH–.

[0365] In some embodiments, the modified amino acid has the structure shown in Formula II-1: Or its salts, wherein V and R both have the definitions described in Formula I-1. In some embodiments according to this paragraph, V is –W 1a –W 2a –;W 1a and W 2a One of them is absent or is –CH2–, and the other is –NH–, –O–, or –S–. In some embodiments, V is a single bond, –NH–, or –CH2NH–. In some embodiments, V is a single bond or –CH2NH–; and R is a methyl group.

[0366] In some embodiments, the modified amino acid has the structure shown in Formula III-1: Or its salts, wherein V and R both have the definitions described in Formula I-1. In some embodiments according to this paragraph, V is –W 1a –W 2a W 1a and W 2a One of them is absent or is –CH2–, and the other is –NH–, –O–, or –S–. In some embodiments, V is a single bond, –NH–, or –CH2NH–. In some embodiments, V is a single bond, –NH–, or –CH2NH–; and R is a methyl group.

[0367] In some embodiments, the modified amino acid has the structure shown in Formula IV-1: Or its salts, wherein V and R both have the definitions described in Formula I-1. In some embodiments according to this paragraph, V is –W 1a –W 2a –;W 1a and W 2a One of them is absent or is –CH2–, and the other is –NH–, –O–, or –S–. In some embodiments, V is a single bond, –NH–, or –CH2NH–. In some embodiments, V is a single bond, –NH–, or –CH2NH–; and R is a methyl group.

[0368] In some embodiments, the modified amino acid has the structure shown in Formula V-1: Or its salts, wherein both V and R have the definitions described in Formula I. In some embodiments according to this paragraph, V is –W 1a –W 2a –;W 1a and W 2a One of them is absent or is –CH2–, and the other is –NH–, –O–, or –S–. In some embodiments, V is a single bond, –NH–, or –CH2NH–. In some embodiments, V is a single bond, –NH–, or –CH2NH–; and R is a methyl group.

[0369] In some embodiments, the modified amino acid has the structure shown in Formula VI-1: Formula VI-1; Or its salts, wherein both V and R have the definitions described in Formula I. In some embodiments according to this paragraph, V is –W 1a –W 2a –;W 1a and W 2a One of them is absent or is –CH2–, and the other is –NH–, –O–, or –S–. In some embodiments, V is a single bond, –NH–, or –CH2NH–. In some embodiments, V is a single bond, –NH–, or –CH2NH–; and R is a methyl group.

[0370] In some embodiments, the modified amino acid has the structure shown in Formula VII-1: Or its salts, wherein both V and R have the definitions described in Formula I. In some embodiments according to this paragraph, V is –W 1a –W 2a –;W 1a and W2a One of them is absent or is –CH2–, and the other is –NH–, –O–, or –S–. In some embodiments, V is a single bond, –NH–, or –CH2NH–. In some embodiments, V is a single bond, –NH–, or –CH2NH–; and R is a methyl group.

[0371] In some embodiments, the modified amino acid has the structure shown in Formula VIII-1: Or its salts, wherein both V and R have the definitions described in Formula I. In some embodiments according to this paragraph, V is –W 1a –W 2a –;W 1a and W 2a One of them is absent or is –CH2–, and the other is –NH–, –O–, or –S–. In some embodiments, V is a single bond, –NH–, or –CH2NH–. In some embodiments, V is a single bond, –NH–, or –CH2NH–; and R is a methyl group.

[0372] In some embodiments, the modified amino acid has the structure shown in Formula IX-1: Formula IX-1; Or its salts, wherein V and R both have the definitions described in Formula I-1. In some embodiments according to this paragraph, V is –W 1a –W 2a –;W 1a and W 2a One of them is absent or is –CH2–, and the other is –NH–, –O–, or –S–. In some embodiments, V is a single bond, –NH–, or –CH2NH–. In some embodiments, V is a single bond, –NH–, or –CH2NH–; and R is a methyl group.

[0373] In some embodiments, the modified amino acid has a structure shown in any of formulas 51-62: Or its salt.

[0374] In any of the foregoing embodiments, if the conjugate has the structure described in this invention, for example, having the structure shown in any of Formulas 1101-1118 and / or Formulas 1201A-1201B, the structure within the brackets may be covalently linked to one or more unnatural or modified amino acids of the antibody, wherein the one or more unnatural or modified amino acids are located at sites selected from the group consisting of: HC-F404, HC-Y180, HC-F241, HC-Y391, LC-K42, and LC-E161 according to the Kabat numbering scheme of the Kabat or EU. In some embodiments, the structure within the brackets is covalently linked to one or more unnatural or modified amino acids located at site HC-F404 of the antibody. In some embodiments, the structure within the brackets is covalently linked to one or more unnatural or modified amino acids located at site HC-F404 of the antibody. In some embodiments, the structure within the brackets is covalently linked to one or more unnatural or modified amino acids located at site HC-Y180 of the antibody. In some embodiments, the bracketed structure is covalently linked to one or more non-natural or modified amino a...

Claims

1. An antibody-drug conjugate comprising an antibody or an antigen-binding fragment thereof, said antibody or antigen-binding fragment being covalently linked via a first linker to at least one topoisomerase inhibitor and via a second linker to at least one DNA damage response (DDR) inhibitor.

2. The antibody conjugate according to claim 1, wherein, The antibody conjugate is shown in the structure of formula (I): Equation (I) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof; Where Ab is the antibody or its antigen-binding fragment; L a It is the first connector; L b It is the second connector; Each DDRi is an independent residue of a DDR inhibitor; Each TOPOi is an independent residue of a topoisomerase inhibitor; The subscript n is an integer selected from 1 to 10; and The subscript m is an integer selected from 1 to 10.

3. The antibody conjugate according to claim 1 or 2, wherein, The topoisomerase inhibitor is a topoisomerase I inhibitor.

4. The antibody conjugate according to claim 3, wherein, The topoisomerase I inhibitor is a camptothecin derivative.

5. The antibody conjugate according to claim 3, wherein, The topoisomerase I inhibitor is selected from the group consisting of: camptothecin, irinotecan, SN-38, topotecan, exatecan, and their derivatives.

6. The antibody conjugate according to claim 5, wherein, The topoisomerase I inhibitor is exatecan.

7. The antibody conjugate according to claim 1 or 2, wherein, The topoisomerase inhibitor is a topoisomerase II inhibitor.

8. The antibody conjugate according to claim 7, wherein, The topoisomerase II inhibitor is selected from etoposide, teniposide, tafluposide, and their derivatives.

9. The antibody conjugate according to any one of the preceding claims, wherein, The DNA damage response inhibitors are poly(ADP-ribose) polymerase (PARP) inhibitors, checkpoint kinase 1 (CHK1) inhibitors, ataxia-telangiectasia and Rad3-associated (ATR) kinase inhibitors, nicotinamide phosphoribosyltransferase 1 (NAMPT1) inhibitors, or RAD51 inhibitors.

10. The antibody conjugate according to claim 9, wherein, The DNA damage response inhibitor is a PARP inhibitor.

11. The antibody conjugate according to claim 10, wherein, The PARP inhibitors are selected from olaparib, veliparib, talazoparib, rucaparib, niraparib, AZD5305, talzenna, and their derivatives.

12. The antibody conjugate according to claim 11, wherein, The PARP inhibitor is talazoparib or a derivative thereof.

13. The antibody conjugate according to any one of the preceding claims, wherein, The first linker and the second linker each independently comprise a protease-cleavable linker, an enzyme-cleavable linker, a pH-sensitive linker, or an uncleavable linker.

14. The antibody conjugate according to any one of the preceding claims, wherein, The first linker and the second linker each independently comprise a β-glucuronidase-cleavable β-glucuronide or a cathepsin-cleavable linker.

15. The antibody conjugate according to any one of claims 2-14, wherein, The antibody conjugate shown in formula (I) is as shown in formula (Ia-1): Equation (Ia-1) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof; wherein: Ab is the antibody or its antigen-binding fragment; Each SP 1 SP 2 and SP 3 Each time it appears, it is either independently absent or independently a divalent spacer group; Each W 1 Each time it appears, it either does not exist independently, or it is independently... Wherein -NH- is connected to W 2 and each These respectively represent the connection points that connect to the rest of the general formula; L 1 Independently as a key, or L 1 C can be substituted independently. 1-6 alkylene, wherein the C 1-6 The alkylene group is optionally substituted by one, two or three substituents selected from halogen, alkyl, haloalkyl, hydroxyl, amino, alkylamino and alkoxy groups; Each W 2 Each time it appears, it is either independently absent or independently an amino acid residue or a peptide residue, wherein the amino acid residue or the peptide residue is optionally represented by HP. 2 Group substitution; Each HP 1 Each time it appears, it is either independently absent or independently a divalent hydrophilic group; Each HP 2 When present, they are monovalent hydrophilic groups; Each R A Each time it appears, it is independently represented by an arbitrarily substituted C. 1-6 Alkyl, wherein the C 1-6 The alkyl group is optionally substituted by one, two or three substituents selected from halogens, alkyl groups, haloalkyl groups, hydroxyl groups, amino groups, alkylamino groups and alkoxy groups; Each RT is either independently absent or independently a release trigger group each time it appears. Each RL is a reactive linker. y is an integer independently selected from 0, 1, and 2; and z is an integer independently selected from 0 and 1.

16. The antibody conjugate according to claim 15, wherein, The antibody conjugate shown in formula (Ia-1) is as shown in formula (Ia): Formula (Ia) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof; wherein: Each W 1 Each time it appears, it either does not exist independently, or it is independently... Wherein -NH- is connected to W 2 ; and each These represent the connection points that connect to the rest of the general formula.

17. The antibody conjugate according to any one of claims 2-16, wherein, The antibody conjugate shown in formula (I) is shown in formula (Ib): Formula (Ib) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof; wherein: L 2 It is -C 1-6 alkylene-, the -C 1-6 The alkylene group is optionally substituted with one or more substituents selected from the following: halogen, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; Y is –X 1 -C 1-6 Alkylene-[X] 1 -C 1-6 [alkylene] a -[X 1 ] b –、–X 1 -C 2-6 imidene-[X] 1 -C 2-6 [Alkenyl] a -[X 1 ] b – or –X 1 -C 2-6 Ethyne-[X] 1 -C 2-6 [Iso-ynyl] a -[X 1 ] b – wherein at least one alkylene, alkenylene, or ynylene group in Y is selected from one or more groups chosen from R. 50 Substituents and Wherein the alkylene, alkenylene, or ynylene group in Y is optionally selected from one or more groups selected from R. 51 Substituents of the substituents; R 50 Yes –C 1-6 Alkylene-X 2 -[C 1-6 [alkylene] c -HP 2 –C 2-6 imide-X 2 -[C 2-6 [Alkenyl] c -HP 2 、or –C 2-6 Ethyne-X 2 -[C 2-6 [Iso-ynyl] c -HP 2 , where R 50 Each alkylene, alkenylene, or ynylene group may optionally be substituted by one or more substituents selected from the following: halogen, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; R 51 Independently selected from halogens, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; X 1 and X 2 Each is independently selected from –N(R) 10 –, –C(O)–, and –N(R)– 10 )C(O)–; R 10 Each time it appears, it is independently selected from H and C. 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; a is an integer selected from 0, 1, 2, and 3; b is an integer selected from 0 and 1; c is an integer selected from 0 and 1; and Su is the hexose form of the monosaccharide.

18. The antibody conjugate according to claim 17, having the structure shown in formula (Ib-A): Formula (Ib-A) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

19. The antibody conjugate according to claim 17, having the structure shown in formula (Ib-B): Formula (Ib-B) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

20. The antibody conjugate according to any one of claims 2-16, wherein, The antibody conjugate shown in formula (I) is shown in formula (Ic): Formula (Ic) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof; wherein: L 2 It is -C 1-6 alkylene-, the -C 1-6 The alkylene group is optionally substituted with one or more substituents selected from the following: halogen, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; Y is –X 1 -C 1-6 Alkylene-[X] 1 -C 1-6 [alkylene] a -[X 1 ] b –、–X 1 -C 2-6 imidene-[X] 1 -C 2-6 [Alkenyl] a -[X 1 ] b – or –X 1 -C 2-6 Ethyne-[X] 1 -C 2-6 [Iso-ynyl] a -[X 1 ] b – wherein at least one alkylene, alkenylene, or ynylene group in Y is selected from one or more groups chosen from R. 50 Substituents and Wherein the alkylene, alkenylene, or ynylene group in Y is optionally selected from one or more groups selected from R. 51 Substituents of the substituents; R 50 Yes –C 1-6 Alkylene-X 2 -[C 1-6 [alkylene] c -HP 2 –C 2-6 imide-X 2 -[C 2-6 [Alkenyl] c -HP 2 、or –C 2-6 Ethyne-X 2 -[C 2-6 [Iso-ynyl] c -HP 2 , where R 50 Each alkylene, alkenylene, or ynylene group may optionally be substituted by one or more substituents selected from the following: halogen, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; R 51 Independently selected from halogens, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; X 1 and X 2 Each is independently selected from –N(R) 10 –, –C(O)–, and –N(R)– 10 )C(O)–; R 10 Each time it appears, it is independently selected from H and C. 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; a is an integer selected from 0, 1, 2, and 3; b is an integer selected from 0 and 1; c is an integer selected from 0 and 1; and Su is the hexose form of the monosaccharide.

21. The antibody conjugate according to claim 20, having the structure shown in formula (Ic-A): Formula (Ic-A) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

22. The antibody conjugate according to claim 20, having the structure shown in formula (Ic-B): Formula (Ic-B) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

23. The antibody conjugate according to any one of claims 2-16, wherein, The antibody conjugate shown in formula (I) is shown in formula (Id): Formula(Id) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof; wherein: L 2 It is -C 1-6 alkylene-, the -C 1-6 The alkylene group is optionally substituted with one or more substituents selected from the following: halogen, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; Y is –X 1 -C 1-6 Alkylene-[X] 1 -C 1-6 [alkylene] a -[X 1 ] b –、–X 1 -C 2-6 imidene-[X] 1 -C 2-6 [Alkenyl] a -[X 1 ] b – or –X 1 -C 2-6 Ethyne-[X] 1 -C 2-6 [Iso-ynyl] a -[X 1 ] b – wherein at least one alkylene, alkenylene, or ynylene group in Y is selected from one or more groups chosen from R. 50 Substituents and Wherein the alkylene, alkenylene, or ynylene group in Y is optionally selected from one or more groups selected from R. 51 Substituents of the substituents; R 50 Yes –C 1-6 Alkylene-X 2 -[C 1-6 [alkylene] c -HP 2 –C 2-6 imide-X 2 -[C 2-6 [Alkenyl] c -HP 2 、or –C 2-6 Ethyne-X 2 -[C 2-6 [Iso-ynyl] c -HP 2 , where R 50 Each alkylene, alkenylene, or ynylene group may optionally be substituted by one or more substituents selected from the following: halogen, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; R 51 Independently selected from halogens, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; X 1 and X 2 Each is independently selected from –N(R) 10 –, –C(O)–, and –N(R)– 10 )C(O)–; R 10 Each time it appears, it is independently selected from H and C. 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups; a is an integer selected from 0, 1, 2, and 3; b is an integer selected from 0 and 1; c is an integer selected from 0 and 1; and Su is the hexose form of the monosaccharide.

24. The antibody conjugate according to claim 23, having the structure shown in formula (Id-A): Formula (Id-A) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

25. The antibody conjugate according to claim 23, having the structure shown in formula (Id-B): Formula (Ic-B) Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

26. The antibody conjugate according to any one of claims 15-125, wherein, SP 2 and SP 3 Each exists independently, or each is independently selected from -C. 1-6 Alkylene -; -C(O)-; -C 1-6 alkylene-C(O)-, wherein the -C(O)- is respectively connected to W 2 or HP 1 ;-C(O)-C 1-6 Alkylene -C(O)-;-C(O)(C 1-6 alkylene)NR 1 C(O)-;-C(O)(C 1-6 Alkylene)OC(O)-; and -C(O)(C 1-6 (alkylene)SC(O)-; where R 1 It is H or C that is optionally substituted. 1-6 Alkyl groups; and SP 2 or SP 3 The C mentioned 1-6 Alkylene, either alone or as part of another group, is optionally substituted with one, two or three substituents selected from halogen, alkyl, haloalkyl, hydroxyl, amino, alkylamino and alkoxy groups, respectively.

27. The antibody conjugate according to claim 26, wherein, SP 3 It does not exist independently, or SP 3 Independently -C(O)-C 1-6 Alkylene-C(O)-.

28. The antibody conjugate according to claim 27, wherein, SP 3 It does not exist.

29. The antibody conjugate according to any one of claims 26-28, wherein, SP 2 It does not exist independently, or SP 2 Independently for -C 1-6 alkylene-C(O)-, wherein the -C(O)- is attached to W 2 .

30. The antibody conjugate according to claim 29, wherein, SP 2 Independently for -C 1-2 Alkylene-C(O)-.

31. The antibody conjugate according to any one of claims 15-30, wherein, HP 1 Independently , where R 2 It is H or methyl, and x1 is an integer between 1 and 50, inclusive; and each These are the connection points that connect to the rest of the compound.

32. The antibody conjugate according to claim 231, wherein, R 2 It is H.

33. The antibody conjugate according to claim 31 or 32, wherein, x1 is an integer between 1 and 10, inclusive.

34. The antibody conjugate according to claim 33, wherein, x1 is an integer between 1 and 5, inclusive of 1 and 5.

35. The antibody conjugate according to claim 34, wherein, x1 is 4.

36. The antibody conjugate according to any one of claims 15-30, wherein, HP 1 It does not exist.

37. The antibody conjugate according to any one of claims 15-36, wherein, W 2 It does not exist.

38. The antibody conjugate according to any one of claims 15-36, wherein, W 2 Independently, it is an amino acid residue or a peptide residue, wherein said amino acid residue or said peptide residue is optionally HP 2 replace.

39. The antibody conjugate according to claim 38, wherein, W 2 Independently, it is a peptide residue, and the peptide residue contains at least one non-natural amino acid, and the peptide residue is optionally converted by HP. 2 replace.

40. The antibody conjugate according to claim 39, wherein, The at least one non-natural amino acid is selected from 3-sulfoalanine, hydroxyproline (Hyp), citrulline (Cit), ornithine (Orn), leucine (Nle), 3-nitrotyrosine, nitroarginine, pyroglutamic acid (Pyr), naphthylalanine (Nal), 2,4-diaminobutyric acid (DAB), methionine sulfoxide, methionine sulfone, 2,3-diaminopropionic acid, and β-alanine.

41. The antibody conjugate according to any one of claims 15-36, wherein, W 2 It is an amino acid residue independently, and the amino acid residue is a β-amino acid.

42. The antibody conjugate according to claim 41, wherein, The β-amino acid is β-alanine.

43. The antibody conjugate according to any one of claims 15-36, wherein, W 2 Independently selected from -(C(O)CHR 3 NR 4 ) a -、-(C(O)CH2CHR 3 NR 4 ) b -、-(C(O)CHR 3 CH2NR 4 ) b -、-(C(O)CH2CHR 3 NR 4 ) b -(C(O)CHR 3 NR 4 ) a -、-(C(O)CHR 3 CH2NR 4 ) b -(C(O)CHR 3 NR 4 ) a -、-(C(O)CHR 3 NR 4 ) a -(C(O)CH2CHR 3 NR 4 ) b -、and-(C(O)CHR 3 NR 4 ) a -(C(O)CHR 3 CH2NR 4 ) b -; Where R 3 Independently, these are amino acid side chain residues, which optionally are HP-dependent. 2 replace; R 4 Independently H or C 1-6 alkyl; a and b are each independent integers between 1 and 10, inclusive; and W 2 The -C(O)- is connected to W 1 .

44. The antibody conjugate according to claim 43, wherein, W 2 Independently -(C(O)CHR 3 NR 4 ) a - 45. The antibody conjugate according to claim 43, wherein, W 2 Independently -(C(O)CH2CHR 3 NR 4 ) b - 46. ​​The antibody conjugate according to claim 43, wherein, W 2 Independently -(C(O)CH2CHR 3 NR 4 ) b -(C(O)CHR 3 NR 4 ) a - 47. The antibody conjugate according to any one of claims 43-46, wherein, R 3 It is an amino acid side chain residue independently selected from valine, citrulline, alanine, glycine, 3-sulfoalanine and 2,3-diaminopropionic acid.

48. The antibody conjugate according to any one of claims 43-47, wherein, R 4 It is H.

49. The antibody conjugate according to any one of claims 43-48, wherein, a is an independent integer between 1 and 3, inclusive of 1 and 3.

50. The antibody conjugate according to any one of claims 43-49, wherein, b is 1.

51. The antibody conjugate according to claim 44, wherein, W 2 It independently has the structure shown in the following formula: ; and each These are the connection points that connect to the rest of the compound.

52. The antibody conjugate according to claim 51, wherein, W 2 It independently has the structure shown in the following formula: 。 53. The antibody conjugate according to claim 43, wherein, W 2 It independently has the structure shown in the following formula: and each These are the connection points that connect to the rest of the compound.

54. The antibody conjugate according to any one of claims 15-53, wherein, HP 2 Independently , where R 2 It is H or methyl, and x2 is an integer between 1 and 50, inclusive; and each These are the connection points that connect to the rest of the compound.

55. The antibody conjugate according to claim 54, wherein, x2 is an integer between 10 and 20, inclusive.

56. The antibody conjugate according to claim 55, wherein, x2 is 12.

57. The antibody conjugate according to any one of claims 15-56, wherein, W 1 It does not exist.

58. The antibody conjugate according to any one of claims 15-56, wherein, W 1 Independently .

59. The antibody conjugate according to any one of claims 15-56, wherein, W 1 Independently , .

60. The antibody conjugate according to claim 58 or 59, wherein, y is 0.

61. The antibody conjugate according to any one of claims 58-60, wherein, z is 0.

62. The antibody conjugate according to any one of claims 58-60, wherein, z is 1.

63. The antibody conjugate according to any one of claims 58-62, wherein, RT is a β-glucuronide that can be cleaved by β-glucuronidase.

64. The antibody conjugate according to claim 63, wherein, RT is ;and It is the connection point that connects to the rest of the compound.

65. The antibody conjugate according to claim 64, wherein, RT is .

66. The antibody conjugate according to any one of claims 15-65, wherein, SP 1 It does not exist.

67. The antibody conjugate according to any one of claims 15-65, wherein, SP 1 It does not exist independently, or SP 1 Independently selected from -NR 5 CH2- and -NR 4 -triaryl-CH2-, where R 5 It is C 1-6 Alkyl-OCH3, and R 4 Is it H or C? 1-6 alkyl.

68. The antibody conjugate according to claim 67, wherein, SP 1 Independently for -NR 5 CH2-.

69. The antibody conjugate according to claim 68, wherein, SP 1 It is -N(CH2CH2OCH3)CH2-.

70. The antibody conjugate according to claim 69, wherein, SP 1 Independently for -NR 4 -Arylene-CH2-.

71. The antibody conjugate according to claim 70, wherein, SP 1 yes ; and each These are the connection points that connect to the rest of the compound.

72. The coupling according to any one of claims 17-71, wherein, Su is ,in This indicates the connection point that connects to the rest of the compound.

73. The coupling according to any one of claims 17-72, wherein, Su is ,in This indicates the connection point that connects to the rest of the compound.

74. The coupling according to any one of claims 17-73, wherein, L 3 It is -C 1-3 Alkylene-.

75. The coupling according to any one of claims 17-74, wherein, Y is –X 1 -C 1-6 Alkylene-[X] 1 -C 1-6 [alkylene] a -X 1 – where at least one alkylene group in Y is selected from one or more of R 50 Substituents are substituted.

76. The coupling according to any one of claims 17-74, wherein, Y is –X 1 -C 1-4 Alkylene-X 1 -C 1-4 Alkylene-X 1 -C 1-4 Alkylene-X 1 – where at least one alkylene group in Y is selected from one or more of R 50 Substituents are substituted.

77. The coupling according to any one of claims 17-74, wherein, Y is –X 1 -C 1-4 Alkylene-X 1 -C 1-4 Alkylene-X 1 – where at least one alkylene group in Y is selected from one or more of R 50 Substituents are substituted.

78. The coupling according to any one of claims 17-77, wherein, R 50 Yes –C 1-6 Alkylene-X 2 -[C 1-6 [alkylene] c -HP 2 , where R 50 Each alkylene group may optionally be substituted by one or more substituents selected from the following: halogen, -CN, -NO2, -OH, -N(R) 10 )2、-C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 2-10 alkenyl, C 2-10 alkynyl group, C 3-12 Carbon rings, heterocycles consisting of 3 to 12 atoms, and C 1-10 Halogenated alkyl groups.

79. The coupling according to any one of claims 17-78, wherein, c is 1.

80. The coupling according to any one of claims 17-79, wherein, HP 2 It includes polyethylene glycol (PEG), methoxy polyethylene glycol (mPEG), poly(propylene glycol) (PPG), copolymers of ethylene glycol and propylene glycol, poly(oxyethylated polyols), poly(enols), poly(vinylpyrrolidone), poly(hydroxyalkyl methylacrylamide), poly(hydroxyalkyl methacrylates), poly(sugars), poly(α-hydroxy acids), poly(vinyl alcohol), polyphosphazenes, polyoxazoline compounds (POZ), and poly( N α-Acryloylmorpholine), polysarcosine, or combinations thereof.

81. The coupling according to claim 80, wherein, HP 2 It contains polyethylene glycol (PEG) or methoxy polyethylene glycol (mPEG).

82. The antibody conjugate according to any one of claims 15-81, wherein, RL is selected independently ; and each These are the connection points that connect to the rest of the compound.

83. The antibody conjugate according to any one of claims 2-82, wherein, The TOPOi is independently selected from or its derivatives; and each These are the connection points that connect to the rest of the compound.

84. The antibody conjugate according to claim 83, wherein, The TOPOi is independently selected from ; or its derivatives.

85. The antibody conjugate according to claim 83, wherein, The TOPOi is independently selected from ; or its derivatives.

86. The antibody conjugate according to any one of claims 2-785, wherein, The DDRi is independently a PARP inhibitor.

87. The antibody conjugate according to claim 86, wherein, The PARP inhibitors are independently selected from and or its derivatives; and each These are the connection points that connect to the rest of the compound.

88. The antibody conjugate according to claim 86, wherein, The PARP inhibitors are independently selected from or its derivatives; and each These are the connection points that connect to the rest of the compound.

89. The antibody conjugate according to any one of claims 2-85, wherein, The DDRi is independently an ATR inhibitor.

90. The antibody conjugate according to claim 89, wherein, The ATR inhibitor is independently selected from or its derivatives; and each These are the connection points that connect to the rest of the compound.

91. The antibody conjugate according to any one of claims 2-85, wherein, The DDRi is independently a NAMPT1 inhibitor.

92. The antibody conjugate according to claim 91, wherein, The NAMPT1 inhibitor is independently selected from or its derivatives; and each These are the connection points that connect to the rest of the compound.

93. The antibody conjugate according to any one of claims 2-85, wherein, The DDRi is independently a CHK1 inhibitor.

94. The antibody conjugate according to claim 93, wherein, The CHK1 inhibitor is independently selected from or its derivatives; and each These are the connection points that connect to the rest of the compound.

95. The antibody conjugate according to any one of claims 15-18, wherein, It has the following structure: ； Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

96. The antibody conjugate according to any one of claims 15-19, wherein, It is selected from the following structures: Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

97. The antibody conjugate according to any one of claims 15-19, wherein, It is selected from the following structures: Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

98. The antibody conjugate according to any one of claims 15-19, wherein, It is selected from the following structures: ， Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

99. The antibody conjugate according to any one of claims 15-19, wherein, It is selected from the following structures: 。 100. The antibody conjugate according to any one of claims 15-16 and 20-22, wherein, It is selected from the following structures: and 。 101. The antibody conjugate according to any one of claims 15-16 and 20-22, wherein, It is selected from the following structures: and .

102. The antibody conjugate according to any one of claims 2-101, wherein, m and n are each independent integers between 1 and 8, inclusive of 1 and 8.

103. The antibody conjugate according to any one of claims 2-101, wherein, n is an integer between 1 and 5, inclusive of 1 and 5.

104. The antibody conjugate according to any one of claims 2-101, wherein, m is an integer between 1 and 5, inclusive of 1 and 5.

105. The antibody conjugate according to any one of claims 2-101, wherein, m + n is at least 10.

106. The antibody conjugate according to any one of claims 2-101, wherein, m + n is at least 12.

107. The antibody conjugate according to any one of claims 2-106, wherein, The molar ratio of DDRi to TOPOi is approximately 1:

1.

108. The antibody conjugate according to any one of claims 2-106, wherein, The molar ratio of DDRi to TOPOi is approximately 2:1 or 1:

2.

109. The antibody conjugate according to any one of claims 1-108, wherein, The antibody or its antigen-binding fragment contains one or more cysteine ​​residues, lysine residues, and / or N-terminal residues covalently linked to the first linker and / or the second linker.

110. The antibody conjugate according to any one of claims 1-108, wherein, The antibody or its antigen-binding fragment comprises: one or more cysteine ​​residues covalently linked to either the first linker or the second linker, and one or more modified amino acids covalently linked to the other of the first linker or the second linker.

111. The antibody conjugate according to any one of claims 1-108, wherein, The antibody or its antigen-binding fragment contains one or more modified amino acids.

112. The antibody conjugate according to claim 111, wherein, The antibody or its antigen-binding fragment comprises one or more modified amino acids, which are covalently linked to the first linker and / or the second linker.

113. The antibody conjugate according to any one of claims 1-108, wherein, (a) the antibody or its antigen-binding fragment is covalently linked to the first linker via a modified amino acid, and (b) the antibody or its antigen-binding fragment is covalently linked to the second linker via a modified amino acid different from that in (a).

114. The antibody conjugate according to any one of claims 110-113, wherein, The one or more modified amino acid residues are independently selected from the group consisting of: ortho-substituted tyrosine, meta-substituted tyrosine, para-substituted phenylalanine, ortho-substituted phenylalanine, and meta-substituted phenylalanine.

115. The antibody conjugate according to claim 114, wherein, The one or more modified amino acid residues are selected from the group consisting of: p-acetyl-L-phenylalanine, O-methyl-L-tyrosine, 3-methyl-phenylalanine, O-4-allyl-L-tyrosine, 4-propyl-L-tyrosine, fluorinated phenylalanine, isopropyl-L-phenylalanine, p-azido-L-phenylalanine, p-acyl-L-phenylalanine, p-benzoyl-L-phenylalanine, p-iodophenylalanine, p-bromophenylalanine, p-amino-L-phenylalanine, isopropyl-L-phenylalanine, p-propoxy-phenylalanine, and p-azidomethyl-L-phenylalanine.

116. The antibody conjugate according to claim 115, wherein, The one or more modified amino acid residues are selected from: p-azidomethyl-L-phenylalanine, p-azido-L-phenylalanine, and p-acetyl-L-phenylalanine.

117. The antibody conjugate according to any one of claims 110-116, wherein, The one or more modified amino acid residues are located at positions selected from the group consisting of the following heavy chain residues or light chain residues: HC-F404, HC-K121, HC-Y180, HC-F241, HC-221, HC-Y391, LC-T22, LC-S7, LC-N152, LC-K42, LC-E161, LC-D170, HC-S136, HC-S25, HC-A40, HC-S119, HC-S190, HC-K222, HC-R19, HC-Y52, or HC-S70, or their translated modified variants, according to the Kabat, Chothia, or EU numbering schemes.

118. The antibody conjugate according to claim 117, wherein, The modified amino acid residues are located at positions selected from the group consisting of the following heavy chain residues or light chain residues: HC-F404, HC-Y180, HC-F241, HC-Y391, and LC-K42.

119. The antibody conjugate according to any one of claims 1-118, wherein, The antibody or its antigen-binding fragment contains p-azidomethyl-L-phenylalanine at HC-F404 and / or HC-241.

120. The antibody conjugate according to any one of claims 1-119, wherein, The antibody or its antigen-binding fragment contains p-acyl-L-phenylalanine at LC-K42.

121. A pharmaceutical composition comprising the antibody-drug conjugate of any one of claims 1-120 and a pharmaceutically acceptable carrier, excipient, or diluent.

122. The compound represented by formula (P-1) or (P-2): Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof; Where R 18 Is it F, or -X? 3 -C 1-4 Alkyl-X 4 ; R 19 It is H; amino C 1-3 Alkyl; C 1-4 Alkylamino C 1-4 Alkyl; C 1-4 Dialkylamino C 1-4 Alkyl; Aryl C 1-4 Alkyl groups, wherein the aryl groups are optionally R 23 Replace; or -C 1-4 Alkyl-X 3 -C 1-4 Alkyl-X 4 ; R 20 Selected from and ; R 22a and R 22b Each was independently selected from C 1-4 Alkyl; amino C 1-3 Alkyl; C 1-4 Alkylamino C 1-4 Alkyl; C 1-4 Dialkylamino C 1-4 Alkyl; and hydroxyl C 1-3 alkyl; R 23 Independently selected from H, amino, amino C 1-3 Alkyl, halogen, C 1-3 Alkyl, hydroxyl, and C 1-3 Alkoxy; R 21a It is F, amino, hydroxyl, -X 3 -C 1-4 Alkyl-X 4 -X 3 -Carbon ring, X 3 - Heterocyclic, amino C 1-3 Alkyl or hydroxyl C 1-3 alkyl; R 21b Selected from H, amino C 1-3 Alkyl and hydroxyl C 1-3 alkyl; R 24a and R 24b Each is independently selected from H and C. 1-4 alkyl; X 3 Is it -O- or -NR? 24a -;and X 4 It is -OH, -OC 1-4 Alkyl, or -NR 24a R 24b ; in, (a) When R 21a When it is F, amino, or hydroxyl, then R 21b Not for H; and (b) when R 19 It is H and R 18 When it is F, then R 22a Not C 1-4 alkyl.

123. The compound according to claim 122, wherein the compound is selected from: Or a pharmaceutically acceptable salt, solvate, stereoisomer, regioisomer, or mixture of regioisomers thereof.

124. A method of treating a disease or condition in a subject in need, comprising administering to the subject an effective amount of the antibody-drug conjugate of any one of claims 1-120, the pharmaceutical composition of claim 121, or the compound of any one of claims 122-123.

125. A method for diagnosing a disease or ailment of a subject in need, comprising administering to the subject an effective amount of an antibody-drug conjugate according to any one of claims 1-120, a pharmaceutical composition according to claim 121, or a compound according to any one of claims 122-123.

126. The method according to claim 124 or 125, wherein, The disease or condition mentioned is cancer, or an inflammatory disease or condition.

127. Use of an effective amount of the antibody-drug conjugate of any one of claims 1-120, the pharmaceutical composition of claim 121, or the compound of any one of claims 122-123 in the preparation of a medicament for treating or preventing a disease or condition in a subject of need.

128. Use of an effective amount of the antibody-drug conjugate of any one of claims 1-120, the pharmaceutical composition of claim 121, or the compound of any one of claims 122-123 in the preparation of a medicament for diagnosing a disease or condition in a subject in need.

129. The method according to claim 127 or 128, wherein, The disease or condition mentioned is cancer, or an inflammatory disease or condition.

130. A kit comprising the antibody-drug conjugate of any one of claims 1-120, the pharmaceutical composition of claim 121, or the compound of any one of claims 122-123, and instructions for use.

131. The kit according to claim 130, wherein, The antibody-drug conjugate or pharmaceutical composition is lyophilized.

132. The kit of claim 131, further comprising a liquid for reconstructing the lyophilized antibody-drug conjugate or pharmaceutical composition.

133. A method for preparing the antibody conjugate according to any one of claims 1-120, comprising: Prepare a conjugate of the light chain (LC) of either the topoisomerase inhibitor or the DNA damage response (DDR) inhibitor with the antibody or its antigen-binding fragment; and The heavy chain (HC) of the antibody or its antigen-binding fragment is conjugated to either the topoisomerase inhibitor or the DNA damage response (DDR) inhibitor.

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