Compositions Comprising Conjugate Therapeutic Enhancers

Abstract

A composition having the structure PNL PM A first compound having an MOI of 1, wherein PN is a protein drug moiety containing a lysine residue, and L PM is a linker and MOI is a moiety of interest, and a second compound having the structure LG-OH, where LG is a group that contains a target binding moiety that binds to a targeting agent.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to Application No. 63 / 189,503, filed May 17, 2021, which is incorporated by reference in its entirety.

[0002] The present disclosure relates to conjugate therapeutic enhancers that are useful for preventing and / or treating various conditions, disorders, or diseases. In particular, the present disclosure relates to protein conjugates, such as antibody-drug conjugates, that can function as therapeutic enhancers. [Background technology]

[0003] Conjugate therapeutic enhancers are widely used to prevent and / or treat various conditions, disorders, and diseases. Such enhancers typically include a therapeutically active molecule, such as an antibody, linked to a moiety that has affinity for a specific target involved in the condition, disorder, or disease. However, most of the known conjugation techniques are not directed to a specific site of the therapeutically active molecule and usually result in a mixture of conjugates. There is still a need in the development of site-specific conjugation techniques that provide reaction products with a high degree of homogeneity. Summary of the Invention

[0004] The present disclosure relates to compositions comprising a therapy enhancer agent that contains a moiety of interest conjugated to a targeting agent moiety at a specific location.

[0005] In one embodiment, the composition comprises: Structure of formula (P-II) PNL PM -MOI(P-II) A first compound having the formula: PN is a protein drug moiety that contains a lysine residue; L PM is the linker, The MOI is the moiety of interest, the first compound, The following structure: LG-OH(LG-I) wherein LG is a group that includes a target binding moiety that binds to a targeting agent.

[0006] In another embodiment, the composition comprises: Formula (RI) LG-RG-L RM -MOI(RI) wherein LG is a group comprising a target binding moiety that binds to a targeting agent and is identical to LG in formula (LG-I); RG is a reactive group, L RM is a linker, the same as in formula (P-II), and MOI is the moiety of interest, a third compound, Formula (R-III) HO-RG-L RM -MOI(R-III) or a fourth compound having the formula: [Brief description of the drawings]

[0007] [Figure 1] Representative UPLC / UV280nm traces are provided for compound I-36 (MATE Reagent I-20 conjugated to IVIG). UPLC conditions are shown in Example 2. uABT indicates universal antibody binding terminal, and DAR is the drug-antibody ratio. [Diagram 2] UPLC traces of unconjugated IVIG (Figure 2A), MATE impurity pool standard (Figure 2B), and I-36 conjugate (Figure 2C). [Diagram 3] Calibration curves for uABT, MATE-linker, and pooled MATE reagents, and individual impurities. Samples are prepared and analyzed by the method given in Example 2. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0008] The following detailed description is provided to assist those skilled in the art in implementing the present invention. Exemplary embodiments are described in detail hereinafter. However, these embodiments are merely exemplary, and the present disclosure is not limited thereto, but rather defined by the scope of the appended claims. Those skilled in the art may make modifications and variations in the embodiments described herein without departing from the spirit or scope of the present disclosure.

[0009] Accordingly, embodiments are described below by reference to structures and schemes merely to illustrate aspects of the specification. As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items. The term "or" means "and / or." A phrase such as "at least one of" preceding a list of elements modifies the entire list of elements and not the individual elements of the list.

[0010] When an element is referred to as being "on" another element, it will be understood that it may be in direct contact with the other element, or there may be intervening elements between them. In contrast, when an element is referred to as being "directly on" another element, there are no intervening elements present.

[0011] Terms such as first, second, third, etc. may be used herein to describe various elements, components, regions, layers, and / or sections, but it should be understood that these elements, components, regions, layers, and / or sections should not be limited by these terms. These terms are used only to distinguish one element, component, region, layer, or section from another element, component, region, layer, or section. Thus, a first element, component, region, layer, or section discussed below can be referred to as a second element, component, region, layer, or section without departing from the teachings of the present embodiment.

[0012] As used herein, the terms "comprises" and / or "comprising", or "includes" and / or "including" specify the presence of stated features, regions, integers, steps, operations, elements, and / or components, but are to be understood as not excluding the presence or addition of one or more other features, regions, integers, steps, operations, elements, components, and / or groups thereof.

[0013] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. The technical terms used herein are intended only to describe specific embodiments and are not intended to be limiting. It is further understood that terms such as those defined in commonly used dictionaries should be interpreted to have a meaning consistent with their meaning in the relevant technical field and the context of this disclosure, and should not be interpreted in an idealized or overly formal sense unless expressly defined herein.

[0014] As used in this application, unless expressly provided herein, each of the following terms shall have the meaning set forth below. Additional definitions are set forth throughout this application. If a term is not specifically defined herein, the term will be given its art-recognized meaning by one of ordinary skill in the art applying the term in that context.

[0015] The articles "a" and "an" refer to one or to more than one (i.e., to at least one) of the grammatical object of the article, unless the context clearly dictates otherwise. By way of example, "an element" means one element or more than one element.

[0016] As used herein, unless a specific definition is provided otherwise, the term "substituted" refers to a group substituted with deuterium, halogen (-F, -Cl, -Br, -I), a hydroxy group (-OH), an amino group (-NH2), a carboxyl group (-CO2H), a substituted or unsubstituted C1-C10 amine group, a nitro group (-NO2), a C1-C10 alkyl group, a C3-C10 cycloalkyl group, a C6-C12 aryl group, a C1-C10 alkoxy group, a C1-C10 trifluoroalkyl group such as a trifluoromethyl group (-CF3), or a cyano group (-CN) in place of at least one hydrogen of a substituent or compound.

[0017] Additional aspects are set forth in part in the description that follows, and in part will be apparent from the description.

[0018] The starting materials useful for making the pharmaceutical compositions of the present disclosure are readily commercially available or can be prepared by one of ordinary skill in the art.

[0019] The present disclosure relates to compositions comprising a therapy enhancer agent that contains a moiety of interest conjugated to a targeting agent moiety at a specific location.

[0020] In one embodiment, the composition comprises: Structure of formula (P-II) PNL PM -MOI(P-II) A first compound having the formula: PN is a protein drug moiety that contains a lysine residue; L PM is the linker, The MOI is the moiety of interest, the first compound, The following structure: LG-OH(LG-I) wherein LG is a group that includes a target binding moiety that binds to a targeting agent.

[0021] In another embodiment, the composition comprises: Formula (RI) LG-RG-L RM -MOI(RI) wherein LG is a group comprising a target binding moiety that binds to a targeting agent and is identical to LG in formula (LG-I); RG is a reactive group, L RM is a linker, and is the same as in formula (P-II), The MOI is the moiety of interest, the third compound, Formula (R-III) HO-RG-L RM -MOI(R-III) or a fourth compound having the formula:

[0022] The above compounds having the structures of formulas (P-II), (LG-I), (RI), and (R-III) are described in detail in International Application No. PCT / US20 / 61127, filed November 18, 2020, the entirety of which is incorporated herein by reference.

[0023] target After reading this disclosure, those skilled in the art will understand that the techniques provided herein are useful for conjugating a variety of targeting agents to many types of moieties of interest. In some embodiments, the techniques provided are particularly useful for conjugating protein agents to a variety of moieties of interest. In some embodiments, the targeting agent is or includes a protein agent, a nucleic acid, or a combination thereof.

[0024] In some embodiments, the targeting agent is or comprises a protein drug. In some embodiments, the targeting agent is a protein drug. In some embodiments, the targeting agent is a native protein in a cell, tissue, organ, or organism. In some embodiments, the targeting agent is an endogenous protein. In some embodiments, the targeting agent is an exogenous protein. In some embodiments, the targeting agent is a manufactured protein, e.g., a protein produced using various biotechnologies. In some embodiments, the targeting agent is an antibody drug. In some embodiments, the targeting agent is an antibody useful as a therapeutic agent. A variety of such antibodies are known in the art and may be utilized as targeting agents. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a polyclonal antibody. In some embodiments, the antibody is an IgG antibody. In some embodiments, the antibody is IVIG (in some embodiments, pooled from healthy donors). In some embodiments, the protein comprises an Fc region. In some embodiments, the antibody comprises an Fc region. In some embodiments, the Fc region comprises a single heavy chain or fragment thereof. In some embodiments, the Fc region comprises two heavy chains or fragments thereof. In some embodiments, the antibody is a human antibody. In some embodiments, the antibody is a chimeric antibody. In some embodiments, the antibody is a humanized antibody. In some embodiments, the antibody is a murine antibody.

[0025] In some embodiments, when characterizing polyclonal antibody or IVIG agents, digestion, e.g., enzymatic digestion using IdeZ, IdeS, etc., is performed either before, during, or after conjugation, resulting in removal of certain regions of the antibody (e.g., the Fab) and providing a composition with improved homogeneity for characterization (e.g., by MS).

[0026] In some embodiments, the antibody is a therapeutic antibody, e.g., an FDA approved antibody for therapeutic use. In some embodiments, the therapeutic antibody is useful for the treatment of cancer. In some embodiments, the antibody is selected from the group consisting of adalimumab, alemtuzumab, atezolizumab, avelumab, ipilimumab, cetuximab, daratumumab, dinutuximab, elotuzumab, ibritumomab tiuxetan, imgatuzumab, infliximab, ipilimumab, necitumumab, obinutuzumab, ofatumumab, pertuzumab, reslizumab, rituximab, trastuzumab, mogamulizumab, AMP-224, FS-102, GSK-2857916, ARGX-111, ARGX-110, AFM-13, APN-301, BI-83 6826, BI-836858, enoblitzumab, otlertuzumab, veltuzumab, KHK-4083, BIW-8962, ALT-803, carotuximab, epratuzumab, inebilizumab, isatuximab, margetuximab, MOR-208, ocaratuzumab, talatuzumab, tremelimumab, benralizumab, lumiliximab, MOR-208, ifibatuzumab, GSK2831781, SEA-CD40, KHK-2823, or BI836858. In some embodiments, the antibody is selected from the group consisting of rituximab, basiliximab, infliximab, cetuximab, siltuximab, dinutuximab, altertoxaximab, daclizumab, palivizumab, trastuzumab, alemtuzumab, omalizumab, efalizumab, bevacizumab, natalizumab, tocilizumab, eculizumab, mogamulizumab, pertuzumab, obinutuzumab, bedrigravir, cefotaxime ... In some embodiments, the antibody is daratumumab. In some embodiments, the antibody is cetuximab.In some embodiments, provided compounds or drugs that comprise an antibody drug moiety are useful for treating a condition, disorder or disease that can be treated by an antibody drug.

[0027] Antibodies may be prepared according to the present disclosure by several techniques. In some embodiments, antibodies may have engineered structures compared to natural immunoglobulins. In some embodiments, antibodies may include specific tags for purification, identification, evaluation, etc. In some embodiments, antibodies may contain fragments (e.g., CDRs and / or Fc, etc.) and may not contain complete immunoglobulins. Those skilled in the art will understand that when antibody sites are cited in the present disclosure (e.g., K246, K248, K288, K290, K317, etc., human antibodies according to EU numbering unless otherwise indicated), the amino acid residue may not be at the exact numbered site, but may be present at the site corresponding to that numbered site and / or sequence homology (e.g., homologs of the same or different species), e.g., according to EU numbering.

[0028] As will be appreciated by those of skill in the art, among other things, the provided techniques can provide for directed conjugation with natural targets (e.g., natural antibodies). In some embodiments, the targeting agent is or comprises a natural antibody drug. In some embodiments, the targeting agent is or comprises an engineered antibody drug. In some embodiments, the targeting agent (e.g., an antibody) does not comprise an engineered non-natural amino acid residue.

[0029] target binding moiety In some embodiments of formulas (LG-I) and (RI), LG is R LG -L LG and R LG teeth, [ka] , R c -(Xaa)z-, a nucleic acid moiety, or a small molecule moiety; each Xaa is independently a residue of an amino acid or amino acid analog; t is 0 to 50; z is 1 to 50; Each R c is independently -L a -R', Each L a are independently a covalent bond or C 20 Aliphatic or C1-C with 1-5 heteroatoms 20 an optionally substituted divalent radical selected from heteroaliphatic, one or more methylene units of the radical are optionally and independently replaced by C(R')-, -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-; Each -Cy- is independently an optionally substituted divalent monocyclic, bicyclic, or polycyclic group, and each monocyclic ring is independently selected from the group consisting of C 3~20 Alicyclic ring, C 6~20 an aryl ring, a 5-20 membered heteroaryl ring having 1-10 heteroatoms, and a 3-20 membered heterocyclyl ring having 1-10 heteroatoms; L LG -L LG1 -, -L LG1 -L LG2 -, -L LG1 -L LG2 -L LG3 - or -L LG1 -L LG2 -L LG3 -L LG4 - and RG is -L RG1 -L RG2 -, -L LG4 -L RG1 -L RG2 -, -L LG3 -L LG4 -L RG1 -L RG2 - or -L LG2 -LLG3 -L LG4 -L RG1 -L RG2 - and L LG1 , L LG2 , L LG3 , L LG4 , L RG1 , L RG2 , and L RM each is independently L; Each L is independently a covalent bond or a divalent optionally substituted straight chain or branched C alkyl group containing one or more aliphatic moieties, aryl moieties, heteroaliphatic moieties each independently having 1 to 20 heteroatoms, heteroaromatic moieties each independently having 1 to 20 heteroatoms, or any combination of any one or more of such moieties. 1~100 is a group, one or more methylene units of the group are optionally and independently selected from C 1~6 Alkylene, C 1~6 Alkenylene, divalent C with 1-5 heteroatoms 1~6 Heteroaliphatic group, -C≡C-, -Cy-, -C(R')2-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')- , -C(O)C(R')2N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S- , -C(O)O-, -P(O)(OR')-, -P(O)(SR')-, -P(O)(R')-, -P(O)(NR')-, -P(S)(OR')-, -P(S)(SR')-, -P( S)(R')-, -P(S)(NR')-, -P(R')-, -P(OR')-, -P(SR')-, -P(NR')-, amino acid residue, or -[(-OC(R')2-C(R')2-) n ]- (wherein n is 1 to 20), each R' is independently -R, -C(O)R, -CO2R, or -SO2R; Each R is independently -H or C 1~30 Aliphatic, C with 1-10 heteroatoms 1~30 Heteroaliphatic, C 6~30Aryl, C 6~30 Arylaliphatic, C with 1-10 heteroatoms 6~30 an optionally substituted group selected from arylheteroaliphatic, 5-30 membered heteroaryl having 1-10 heteroatoms, and 3-30 membered heterocyclyl having 1-10 heteroatoms; or two R groups, optionally and independently, taken together form a covalent bond, or two or more R groups on the same atom optionally and independently, taken together with the atom, form an optionally substituted 3-30 membered monocyclic, bicyclic, or polycyclic ring having, in addition to the atom, 0-10 heteroatoms; or Two or more R groups on two or more atoms optionally and independently, taken together with their intervening atoms, form an optionally substituted 3-30 membered monocyclic, bicyclic, or polycyclic ring having, in addition to the intervening atoms, 0-10 heteroatoms.

[0030] In some embodiments, the LG is or comprises a target binding moiety that binds to a targeting agent, and the targeting agent is an antibody agent.

[0031] In some embodiments, LG is or comprises a target binding moiety that binds to the Fc region, and / or R LG is or comprises DCAWXLGELVWCT (SEQ ID NO:1), wherein the two cysteine ​​residues optionally form a disulfide bond, and X is an amino acid residue.

[0032] In some embodiments, LG is or includes a target binding moiety having a structure of Formulas A-1 through A-50. [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka]

[0033] Purpose Those of skill in the art reading this disclosure will appreciate that various types of object moieties can be utilized for various purposes in accordance with the disclosure.

[0034] In some embodiments, the moiety of interest is or comprises a detectable moiety. Such moieties may be useful for, among other things, detection, quantification, diagnosis, treatment, and the like. In some embodiments, the moiety of interest is or comprises a radioactive label. In some embodiments, the moiety of interest is or comprises a label that can be detected by spectroscopy. In some embodiments, the moiety of interest is or comprises a fluorophore, such as a FITC moiety.

[0035] The moiety of interest may be a moiety that has affinity for a particular target involved in a medical condition, disorder, or disease. In some embodiments, the moiety of interest is or comprises a therapeutic drug moiety. In some embodiments, the moiety of interest is or comprises a drug moiety, e.g., a drug moiety in an antibody-drug conjugate. In some embodiments, the moiety of interest is or comprises a toxic drug. In some embodiments, the moiety of interest is or comprises a cytotoxic drug. In some embodiments, the moiety of interest is or comprises an anti-cancer drug. In some embodiments, the anti-cancer drug is a chemotherapeutic drug.

[0036] In some embodiments, the moiety of interest is or includes a moiety that can interact with and / or recruit other agents, such as proteins, nucleic acids, cells, etc. In some embodiments, the moiety of interest interacts with proteins expressed by a particular cell type, e.g., immune cells, diseased cells, etc. In some embodiments, the moiety of interest is an immune cell binding agent. In some embodiments, the moiety of interest recruits immune cells. In some embodiments, the moiety of interest induces, promotes, and / or enhances one or more immune activities, e.g., to eliminate, kill, and / or inhibit a desired target (e.g., cancer cells, antigens, etc.). In some embodiments, the moiety of interest interacts with, recruits, and / or binds to diseased cells and induces, promotes, and / or enhances the elimination, killing, and / or inhibition of diseased cells.

[0037] In some embodiments, the moiety of interest is or comprises a small molecule drug (e.g., capable of specifically binding to its protein target, cellular target, etc.). In some embodiments, the moiety of interest is or comprises a peptide or protein drug (e.g., scFv, peptide binders to specific targets, etc.). In some embodiments, the moiety of interest is or comprises a nucleic acid drug (e.g., oligonucleotides, mRNA, etc.). In some embodiments, the moiety of interest is or comprises a carbohydrate drug. In some embodiments, the moiety of interest is or comprises a lipid drug.

[0038] In some embodiments, the moiety of interest is or comprises a protein complex (e.g., a Fab). In some embodiments, the moiety of interest is or comprises a fluorophore. In some embodiments, the moiety of interest is or comprises a cytotoxic small molecule drug. In some embodiments, the moiety of interest is or comprises a cytotoxic peptide drug.

[0039] In some embodiments, the moiety of interest is an adjuvant. Those skilled in the art will understand that various adjuvants can be used as the moiety of interest according to the present disclosure. In some embodiments, the adjuvant is one described in US2019 / 0015516, the entirety of which is incorporated herein by reference. In some embodiments, the moiety of interest stimulates the immune system.

[0040] In some embodiments, the moiety of interest is or comprises a particle, hi some embodiments, the particle is or comprises a nanoparticle.

[0041] In some embodiments, the moiety of interest is or comprises a nucleic acid moiety. In some embodiments, the moiety of interest is or comprises an oligonucleotide. In some embodiments, the moiety of interest is or comprises an aptamer.

[0042] In some embodiments, the moiety of interest is an antibody drug. In some embodiments, the moiety of interest is or comprises an antibody fragment. In some embodiments, the moiety of interest is an antibody drug moiety that does not contain the region to which the target binding moiety binds. In some embodiments, the moiety of interest is an antibody drug that does not contain an Fc region. In some embodiments, the moiety of interest is or comprises an scFv. In some embodiments, the scFv is for a different antigen than the antibody targeting drug.

[0043] In some embodiments, the moiety of interest is or comprises a reactive moiety, particularly a reaction partner for bioorthogonal reactions. Suitable reactive moieties, including those for bioorthogonal reactions, are widely known in the art and can be utilized herein. In some embodiments, the bioorthogonal reaction is a cycloaddition reaction, e.g., click chemistry. In some embodiments, the moiety of interest is or comprises -N3. In some embodiments, the moiety of interest is or comprises an alkyne.

[0044] In some embodiments, the moiety of interest may be a moiety that binds to the SARS-CoV-2 virus, which is involved in COVID-19 disease. For example, the moiety that binds to the SARS-CoV-2 virus may be a polypeptide disclosed in L. Cao et al., “De novo design of picomolar SARS-CoV-2 mini-protein inhibitors” Science 370, 426-431 (2020), which is incorporated by reference in its entirety. Such a polypeptide moiety may cause binding to the SARS-CoV-2 spike protein, inhibition, reduction and prevention of binding and / or infection of cells, inhibition, killing and elimination of the SARS-CoV-2 virus and / or cells infected thereby, etc. Various moieties of interest that interact with the SARS-CoV-2 virus are described in International Patent Application No. PCT / US21 / 24186, filed March 25, 2021, U.S. Provisional Patent Application No. 63 / 146584, filed February 6, 2021, and U.S. Provisional Patent Application No. 63 / 182098, filed April 30, 2021, each of which is incorporated by reference in its entirety herein.

[0045] In some embodiments, the moiety of interest improves one or more properties and / or activities of the targeted agent. In some embodiments, the moiety of interest is or includes a stability enhancer. In some embodiments, the moiety of interest improves one or more pharmacodynamic and / or pharmacokinetic properties of the targeted agent.

[0046] In some embodiments, the following conditions (a) the moiety of interest is or contains a therapeutic agent; (b) the moiety of interest is or comprises a moiety capable of binding to a protein, a nucleic acid, or a cell; and / or (c) the moiety of interest is or contains a reactive moiety suitable for a bioorthogonal reaction.

[0047] In some embodiments, the MOI is or comprises a therapeutic drug moiety and / or the MOI is or comprises an antibody drug.

[0048] linking group In some embodiments, the moieties are optionally connected to one another via a linker moiety. For example, in some embodiments, the reactive group (e.g., RG) is linked to a linker (e.g., L RM In some embodiments, the moiety (e.g., LG) is also connected to the moiety of interest (e.g., MOI) via one or more linkers, e.g., L, for linking to various moieties. LG1 , L LG2 , L LG3 , L LG4 In some embodiments, L LG is a linker moiety as described herein. In some embodiments, L LG1 is a linker moiety as described herein. In some embodiments, L LG2 is a linker moiety as described herein. In some embodiments, L LG3 is a linker moiety as described herein. In some embodiments, L LG4 is a linker moiety as described herein. In some embodiments, L RM is a linker moiety as described herein. In some embodiments, L PM is L, as described herein. In some embodiments, L PM is a linker moiety as described herein. In some embodiments, L PM is L as described herein.

[0049] Linker moieties of various types and / or for various purposes (such as those utilized in antibody-drug conjugates) may be utilized in accordance with the present disclosure.

[0050] Linker moieties can be either bivalent or multivalent depending on how they are used. In some embodiments, the linker moiety is bivalent. In some embodiments, the linker is multivalent and connects more than two moieties.

[0051] In some embodiments, L LM is one or more -[(CH2) n -O] m -, wherein each n is independently 1 to 20, and m is 1 to 100.

[0052] In some embodiments, L RM The linker consists of one or more -[(CH2) n -O] m -, wherein each n is independently 1 to 20, and m is 1 to 100.

[0053] Reactive Groups In some embodiments, provided compounds (e.g., those useful as reaction partners) include a reactive group (e.g., RG). As exemplified herein, in many embodiments, in provided compounds, the reactive group (e.g., RG) is located between a first group (e.g., LG) and a moiety of interest (e.g., MOI), and is optionally and independently linked to the first group and the moiety of interest via a linker. In some embodiments, RG is a reactive group as described herein.

[0054] In some embodiments, as shown herein, reactive groups, when utilized in compounds that do not include a target binding moiety, react slowly, resulting in low levels of conjugation of the moiety of interest with the targeting agent, and in some embodiments, substantially no conjugation. As shown herein, the combination of a reactive group and a target binding moiety in the same compound, such as in a compound of formula RI or a salt thereof, can, among other things, facilitate the reaction between the reactive group and the targeting agent, enhance the reaction efficiency, reduce side reactions, and / or improve the reaction selectivity (e.g., with respect to the target site at which conjugation of the moiety of interest with the targeting agent occurs).

[0055] The reactive groups in the provided compounds can react with various types of groups in the targeting agent. In some embodiments, the reactive groups in the provided compounds selectively react with amino groups of the targeting agent, for example, the -NH2 group on the side chain of a lysine residue of a protein. In some embodiments, the reactive groups, when utilized in the provided compounds (e.g., those of formula RI or salts thereof), selectively react with specific sites of the targeting agent, for example, one or more of K246, K248, K288, K290, K317, etc. of IgG1, K251, K253, etc. of IgG2, and K239, K241, etc. of IgG4, as shown in the examples herein. In some embodiments, the site is K246 or K248 of the antibody heavy chain. In some embodiments, the site is K246 and / or K248 of the antibody heavy chain. In some embodiments, the site is K246 of the antibody heavy chain. In some embodiments, the site is K248 of the antibody heavy chain. In some embodiments, the site is K288 or K290 of the antibody heavy chain. In some embodiments, the site is K288 of the antibody heavy chain. In some embodiments, the site is K290 of the antibody heavy chain. In some embodiments, the site is K317. In some embodiments, the site is K414 of the antibody heavy chain. In some embodiments, the site is K185 of the antibody light chain. In some embodiments, the site is K187 of the antibody light chain. In some embodiments, the site is K251 and / or K253 of an IgG2 heavy chain. In some embodiments, the site is K251 of an IgG2 heavy chain. In some embodiments, the site is K253 of an IgG2 heavy chain. In some embodiments, the site is K239 and / or K241 of an IgG4 heavy chain. In some embodiments, the site is K239 of an IgG4 heavy chain. In some embodiments, the site is K241 of an IgG4 heavy chain. In some embodiments, conjugation occurs preferentially at one or more heavy chain sites over light chain sites. In some embodiments, for technologies that do not have a target binding moiety, conjugation occurs at light chain sites rather than heavy chain sites (see, e.g., Figure 15).

[0056] In some embodiments, the reactive group (e.g., RG) is or includes an ester group. In some embodiments, the reactive group (e.g., RG) is or includes an electrophilic group (e.g., a Michael acceptor).

[0057] In some embodiments, RG is of the formula -L LG2 , -L LG2 -L LG3 -L LG4 -L RG1 - or -L RG1 -L RG2 - is a base L LG2 -NH-, -NHC(O)-, -(CH2) n -NHC(O)-, -(CH2) n -OC(O)-, -(CH2) n -OC(O)NH-, -C(O)-NHCH2-, -C(O)-NHCH2CH2-, -C(O)O-CH2-, or NH-C(O)O-CH2-; L LG3 is an optionally substituted aryl ring; L LG4- is a bond, -NH- or -O-; L RG1 is -OC(O)-, -C(O)-, -S(O)-, -OS(O)2-, or -OP(O(OR)-, L RG2 is -CH2-C(O)-, -C(O)-, or -CH2-, L LG is -(O)C-[(CH2) n O] m (CH2) n NH-, -(O)C-[(CH2) n O] m (CH2) n NH-, -[(CH2) n O] m NHC(O)[(CH2) n O] m NH-, -[(CH2) n O] m {NHC(O)[(CH2) n O] m}p NH-, -[(CH2) n O] m Cy[(CH2) n O] m NH-, -[(CH2) n O] m Cy[(CH2) n O] m NHC(O)[(CH2) n O] m NH- or -[(CH2) n O] m Cy[(CH2) n O] m {NHC(O)[(CH2) n O] m} p NH-, wherein n, m, and p at each occurrence are independently selected from 1 to 12, and Cy is an optionally substituted cyclic group.

[0058] In some embodiments, RG is of the formula -L LG2 -L LG3 -L LG4 -L RG1 Based on [ka] is selected from.

[0059] In some embodiments, n is 2 at each occurrence.

[0060] In some embodiments, the reactive group is or includes -C(O)-O- or -OC(O)-.

[0061] In some embodiments, the reactive group comprises an aryl group optionally linked to -C(O)-O- or -OC(O)- and substituted with one or more electron-withdrawing groups.

[0062] In some embodiments, the aryl group is [ka] wherein R s is independently selected at each occurrence from halogen, -NO2, -F, -L-R', -C(O)-L-R', -S(O)-L-R', -S(O)2-L-R', and -P(O)(-L-R')2, where R' is H or C1-C6 alkyl.

[0063] composition In some embodiments, the composition may include an equimolar amount of the first compound and the second compound. In some embodiments, the amount of the second compound may be 50 mole percent (mole%) or less, based on the total number of moles of the first compound and the second compound in the composition. In some embodiments, the amount of the second compound may be 50 mole% or less, 45 mole% or less, 40 mole% or less, 35 mole% or less, 30 mole% or less, 25 mole% or less, 20 mole% or less, 15 mole% or less, 10 mole% or less, or 5 mole% or less, based on the total number of moles of the first compound and the second compound in the composition. In some embodiments, the amount of the second compound may be 5% or less, 4% or less, 3% or less, 2% or less, or 1% or less, based on the total number of moles of the first compound and the second compound in the composition. In some embodiments, the amount of the second compound may be 1.0% or less, 0.9% or less, 0.8% or less, 0.7% or less, 0.6% or less, 0.5% or less, 0.4% or less, 0.3% or less, 0.2% or less, or 0.1% or less based on the total number of moles of the first compound and the second compound in the composition. In some embodiments, the amount of the second compound may be 0.10% or less, 0.09% or less, 0.08% or less, 0.07% or less, 0.06% or less, 0.05% or less, 0.04% or less, 0.03% or less, 0.02% or less, or 0.01% or less based on the total number of moles of the first compound and the second compound in the composition. In some embodiments, the amount of the second compound may be 0.010% or less, 0.009% or less, 0.008% or less, 0.007% or less, 0.006% or less, 0.005% or less, 0.004% or less, 0.003% or less, 0.002% or less, 0.001% or less based on the total number of moles of the first compound and the second compound in the composition. In some embodiments, the amount of the second compound may be 0.0010% or less, 0.0009% or less, 0.0008% or less, 0.0007% or less, 0.0006% or less, 0.0005% or less, 0.0004% or less, 0.0003% or less, 0.0002% or less, 0.0001% or less based on the total number of moles of the first compound and the second compound in the composition.In some embodiments, the amount of the second compound may be 0.00010% or less, 0.00009% or less, 0.00008% or less, 0.00007% or less, 0.00006% or less, 0.00005% or less, 0.00004% or less, 0.00003% or less, 0.00002% or less, or 0.00001% or less based on the total moles of the first compound and the second compound in the composition. In some embodiments, the amount of the second compound may be 0.000010% or less, 0.000009% or less, 0.000008% or less, 0.000007% or less, 0.000006% or less, 0.000005% or less, 0.000004% or less, 0.000003% or less, 0.000002% or less, or 0.000001% or less based on the total moles of the first compound and the second compound in the composition.

[0064] In some embodiments, the composition may further include a third compound, a fourth compound, or a combination thereof. In some embodiments, the amount of the third compound, the fourth compound, or a combination thereof may be 5% or less, 4% or less, 3% or less, 2% or less, or 1% or less based on the moles of the first compound in the composition. In some embodiments, the amount of the third compound, the fourth compound, or a combination thereof may be 1.0% or less, 0.9% or less, 0.8% or less, 0.7% or less, 0.6% or less, 0.5% or less, 0.4% or less, 0.3% or less, 0.2% or less, or 0.1% or less based on the moles of the first compound in the composition. In some embodiments, the amount of the third compound, the fourth compound, or a combination thereof, based on the number of moles of the first compound in the composition, may be 0.10% or less, 0.09% or less, 0.08% or less, 0.07% or less, 0.06% or less, 0.05% or less, 0.04% or less, 0.03% or less, 0.02% or less, 0.01% or less. In some embodiments, the amount of the third compound, the fourth compound, or a combination thereof, based on the number of moles of the first compound in the composition, may be 0.010% or less, 0.009% or less, 0.008% or less, 0.007% or less, 0.006% or less, 0.005% or less, 0.004% or less, 0.003% or less, 0.002% or less, 0.001% or less. In some embodiments, the amount of the third compound, the fourth compound, or a combination thereof may be 0.0010% or less, 0.0009% or less, 0.0008% or less, 0.0007% or less, 0.0006% or less, 0.0005% or less, 0.0004% or less, 0.0003% or less, 0.0002% or less, 0.0001% or less based on the number of moles of the first compound in the composition. In some embodiments, the amount of the third compound, the fourth compound, or combinations thereof may be 0.00010% or less, 0.00009% or less, 0.00008% or less, 0.00007% or less, 0.00006% or less, 0.00005% or less, 0.00004% or less, 0.00003% or less, 0.00002% or less, 0.00001% or less based on the number of moles of the first compound in the composition.In some embodiments, the amount of the third compound, the fourth compound, or combinations thereof may be 0.000010% or less, 0.000009% or less, 0.000008% or less, 0.000007% or less, 0.000006% or less, 0.000005% or less, 0.000004% or less, 0.000003% or less, 0.000002% or less, 0.000001% or less based on the number of moles of the first compound in the composition.

[0065] In another embodiment, a composition comprising: Structure of formula (P-II) PNL PM -MOI(P-II) A first compound having the formula: PN is a protein drug moiety that contains a lysine residue; L PM is the linker, The MOI is the moiety of interest, the first compound, The following structure: LG-OH(LG-I) wherein LG is a group that includes a target binding moiety that binds to a targeting agent; and Formula (RI) LG-RG-L RM -MOI(RI) wherein LG is a group comprising a target binding moiety that binds to a targeting agent and is identical to LG in formula (LG-I); RG is a reactive group, L RM is a linker, and is the same as in formula (P-II), A composition is provided in which the MOI comprises at least one third compound that is a moiety of interest.

[0066] In another embodiment, the composition comprises: Formula (R-III) HO-RG-L RM -MOI(R-III) or a fourth compound having the formula:

[0067] The present invention is further illustrated by the following non-limiting examples. EXAMPLES

[0068] Example 1. Specific techniques for preparing the following agents I-29, I-30, I-31, I-32, I-33, I-34, I-35, I-36. In some embodiments, the disclosure provides techniques for preparing MATE agents and compositions thereof. In some embodiments, the techniques provided include reacting a composition comprising a plurality of antibody agents (e.g., an IVIG composition such as Gamunex-C) with a composition comprising a plurality of agents (e.g., I-3, I-7, I-8, I-15, I-19, I-20, I-21, I-22, I-23, etc.), each of which comprises a target binding moiety, an antibody binding moiety, and a reactive group therebetween (and optional linker moieties linking such moieties). Described below by way of example are preparations of certain MATE agents and compositions thereof.

[0069] 1. Reaction / Conjugation Protocol. 1.1 Exchange of IVIG buffer Gamunex-C (1 mL, approximately 100 mg / mL) was buffer exchanged into 50 mM borate pH 8.2 using an Amicon-15 mL unit with MWCO 30 kDa. The protein concentration of the buffer exchanged IVIG was determined by UV-Vis using an extinction coefficient of 1.41 mL mg-1 cm-1 at 280 nm. Buffer exchanged IVIG was adjusted to 20 mg / mL using 50 mM borate buffer pH 8.2.

[0070] 1.2 Preparation of reagent stock solutions Reagents are weighed out individually and the DMSO volumes used to prepare stock solutions are calculated as follows: DMSO volume (mL) = (solid weight (mg) / molecular weight) x purity (%) / 5mM x 10 6 .

[0071] Data regarding the specific preparation of the reagents is given below. [Table 1]

[0072] The structures of reagents I-7, I-8, I-15, I-19, I-20, I-21, I-22, and I-23 are shown below. [ka] [ka] [ka] [ka]

[0073] 1.3 Setting up the conjugation reaction Instructions for I-29, I-30, I-31, I-32, I-34, I-35, I-36: To IVIG in 50 mM borate pH 8.2 (4 mg, 20 mg / mL, 200 μL) was added the reagent (5 mM in DMSO, 13.3 μL, 2.5 equiv.) The reaction was mixed thoroughly and rotated overnight at room temperature.

[0074] Instructions for I-33: To IVIG in 50 mM borate pH 8.2 (2.62 mg, 20 mg / mL, 130.8 μL) was added reagent (5 mM in DMSO, 8.7 μL, 2.5 equivalents). The reaction was mixed thoroughly and rotated overnight at room temperature. The following table shows the MATE reagents used to generate each MATE reagent, which are described in further detail in the following sections. [Table 2]

[0075] In each of the above procedures, in addition to agents I-29, I-30, I-31, I-32, I-33, I-34, I-35, I-36, the reaction mixture contained the following products formed in equimolar amounts with agents I-29, I-30, I-31, I-32, I-33, I-34, I-35, and I-36. [ka]

[0076] 2. Purification The crude mixture was buffer exchanged into 50 mM glycine pH 2.2 in an Amicon-4mL MWCO 30kDa for 2 cycles. It was then buffer exchanged into PBS in an Amicon-4mL MWCO 30kDa for 2 cycles. An insoluble white precipitate was observed at the bottom of the filter. In some embodiments, the released antibody binding moieties were removed under acidic conditions.

[0077] The solutions and precipitates from the eight samples were each collected in 1.5 mL tubes and centrifuged at 16000 g for 5 min. The clear supernatant was carefully collected. The concentration of the protein conjugate in the supernatant was determined by a UV-Vis Nanodrop instrument using an extinction coefficient of 1.41 mL mg-1 cm-1. The concentration, volume, and yield of the protein conjugate from the specific preparations are shown in the table below. [Table 3]

[0078] To adjust the final MATE drug concentration to a range of 1.5-3.0 mg / mL, all conjugates were diluted with PBS, targeting 3.0 mg / mL. In some embodiments, upon addition of PBS, a white precipitate was observed to rapidly form in some or all samples. To recover the conjugates, the samples were centrifuged at 16000 g for 3 min. The clear supernatant was carefully collected. The concentration of the protein conjugate in the supernatant was determined by a UV-Vis Nanodrop instrument using an extinction coefficient of 1.41 mL mg-1 cm-1. The final conjugate concentration, volume, and yield from the specific preparations are summarized in the table below. [Table 4]

[0079] 3. Analytical methods for determining DAR (target binding moiety / antibody moiety ratio) MATE conjugate (50 μg) was incubated for 1 h at 37° C. with 10× glycerol buffer 2 (0.1× total volume), 1 μL IdeZ and 1 μL PNGase F. An 8 μg sample was then removed and diluted 50-fold with water. Injection volumes were 2 μL or 3 μL depending on the desired signal. device: LC: Waters Acquity UPLC Protein BEH C4 column (300 Å, 17 μm × 2.1 mm × 50 mm) MS: Waters Xevo G2-QTOF Processing software: MassLynx Transfer A: Water + 0.1% formic acid Transfer B: Acetonitrile + 0.1% formic acid

[0080] Data analysis: A total ion chromatogram (TIC) is obtained. For spectral analysis, a TIC region is selected. The region is selected to encompass both conjugated and unconjugated Fc without bias. An MS profile is obtained from the TIC. For deconvolution, a charge state envelope is selected. The raw spectrum is deconvoluted using the MaxEnt1 deconvolution tool to obtain a zero-charge spectrum. The BAR (binder distribution ratio in Fc x Fc per antibody) is calculated using the following formula: BAR=[1(Fc コンジュゲート) ) / [1(Fc コンジュゲート )+1(Fc 非コンジュゲート )] x 2

[0081] Deconvolute at m / z 8000-30000 with a resolution of 1.50. The results of DAR by LC-MS analysis are shown below. [Table 5]

[0082] Throughout this application, various publications are referenced by author name and date, or patent or patent publication number. The disclosures of these publications are incorporated by reference in their entireties into this application in order to more fully describe the state of the art known to those skilled in the art as of the date of the invention described and claimed herein. However, the citation of a reference herein should not be construed as an admission that such reference is prior art.

[0083] Those skilled in the art will recognize or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific procedures described herein. Such equivalents are considered to be within the scope of this disclosure and are covered by the following claims. For example, pharma- ceutically acceptable salts other than those specifically disclosed in the description and examples herein may be used. Furthermore, it is contemplated that specific items in the list of items, or subsets of items within larger groups of items, may be combined with other specific items, subsets of items, or larger groups of items, regardless of the presence or absence of a specific disclosure herein identifying such combinations.

[0084] Example 2. Analytical methods for determining Drug-to-Antibody Ratio (DAR), DAR distribution, and MATE Reagent-related impurities a. Quantification of DAR species and identification of impurities The following UPLC analytical method allows for the simultaneous determination of species with different DARs and MATE reagent-related impurities. Drug substrate / drug product (DR / DP) and DAR / DAR distribution are calculated based on the HPLC / UV280nm signal areas of the individual DAR components. MATE reagent-related DS / DP impurity content (MATE reagent, MATE linker and universal antibody binding terminus (uABT)) is determined as the ratio of sample impurity concentration to DS / DP protein concentration (reported as wt% impurity / wt% protein). The identity of the UPLC signals was confirmed by RS and LCMS analysis of the protein and individual impurities. A representative UPLC / UV280 trace is shown for compound I-36 in Figure 1. DS is the desired product (MATE) and impurities are expressed as wt% of that product (DS / DP).

[0085] The UPLC method is performed on a WATERS ACQUITY UPLC system, UV 280 nm detection. The MS detector is a WATERS XEVO G2-XS QTof detector. This detector is used for molecular weight determination of the species. The autosampler works at ambient conditions of 22-23 °C. A HALO 1000 Å Diphenyl column, 2.7 μm, 2.1 x 50 mm, with a column temperature of 80 °C is used. The flow rate is 0.5 mL / min. Mobile phase A is 0.5% trifluoroacetic acid and mobile phase B is acetonitrile with 0.05% trifluoroacetic acid. [Table 6]

[0086] The MS detector settings on a Waters Xevo G2-XS QTof detector for the impurity analysis and DAR analysis methods are shown in Table 2. [Table 7]

[0087] Dilute DS / DP samples to a protein concentration of approximately 1.5 mg / mL, corresponding to a 50-fold DS / DP dilution in DMSO:water dilution vehicle (6:94 v / v). Samples such as I-36 were stored at -20°C. Thaw samples and equilibrate to ambient temperature for approximately 2 hours. Vortex samples for 1-2 minutes. Dilute to target concentration (1.5 mg / mL protein) in dilution vehicle (DMSO:water = 6:94). Diluted samples are stored at room temperature until analysis, which should be completed within 4 hours of sample dilution. Alternatively, diluted samples may be stored at 2-8°C for 24 hours before analysis. If diluted samples are stored at 2-8°C, samples should be equilibrated to room temperature for 30 minutes and then vortexed for 1-2 minutes before use.

[0088] bI-36 Drug-to-Antibody Ratio (DAR) and DAR Distribution Analysis Determine the DAR and DAR distribution for the I-36 preparation using the UPLC method, detector parameters, and sample preparation methods shown in the previous section. Prior to quantifying the DAR species, obtain UPLC spectra of pooled standards of (i) unconjugated IVIG, (ii) MATE impurities including unconjugated IVIG. These spectra are shown in Figures 2A-C. The data are shown in Tables 3 and 4. [Table 8] [Table 9]

[0089] b. UPLC method for MATE impurity analysis (1) Preparation of calibration curve Prepare pooled standard stock solutions (PL stocks) as follows: Prepare individual uABT, MATE Reagent (MR) and MATE-Linker Standard (STD) stock solutions in DMSO at a STD stock solution concentration of 2.00 mg / mL. Combine equal volumes of individual 1 mg / mL DMSO stock solutions. The resulting individual pooled stock solution concentration is 0.667 mg / mL (PL stock). Store DMSO standard stock solutions at -70°C for up to 1 month.

[0090] To quantify most impurities formed during the MATE reaction, a calibration curve with individual impurity concentration ranges from 0.004mg / mL to 0.04mg / mL (or 0.02ug / injection to 0.2ug / injection for a 5uL injection volume) is required. Thaw the DMSO solution at ambient temperature and then vortex thoroughly. IVIG working stock (82.5mg / mL) is prepared by diluting 165mg / mL IVIG reference material 2-fold with water. Calibration curve criteria follow Table 5. A DMSO concentration of 6% and a constant IVIG:pool impurity ratio (w / w) are maintained for all calibrators. Store the calibrator solutions at ambient temperature. Due to the limited stability of the calibration solutions, measurements should be performed within 4 hours after calibrator preparation. [Table 10]

[0091] (2) Handling of DS / DP samples The diluted DS / DP samples are stable for at least 2 hours. Following the reproducibility study of the diluted samples shown in Table 6, a 50-fold DS / DP sample dilution was selected for impurity analysis. [Table 11]

[0092] The calibration curves were found to be reproducible for individual and pooled impurities. Calibration curve data for uABT, MATE-Linker, and MATE Reagent samples are shown in Figure 3. Calibration curve calculation factors for I-36 / I-20 uABT, MATE-Linker, and MATE Reagent were extracted from the calibration curves and are presented in Table 7. [Table 12]

[0093] Impurities were calculated for duplicate I-36 conjugation reactions using the calibration curve calculation factors shown in Table 7. Results are summarized in Table 8. Total protein concentrations were established using the protein calibration curve. [Table 13]

Claims

1. 1. A composition comprising: Formula (P-II): P-N-L ＰＭ (a) [During the ceremony, P-N is a protein drug moiety containing a lysine residue; L ＰＭ is the linker, MOI is the moiety of interest. or a salt thereof, structure: LG-OH (LG-I) where LG is a group that contains a target binding moiety that binds to a targeting agent. or a salt thereof; and A composition comprising:

2. Formula (RI): LG-RG-L ＲＭ (a) [During the ceremony, LG is a group containing a target binding moiety that binds to a targeting agent, and is the same as LG in formula (LG-I); RG is a reactive group, L ＲＭ is a linker and is the same as L RM in formula (P-II), MOI is the moiety of interest. or a salt thereof; or Formula (R-III): HO-RG-L ＲＭ -MOI(R---I) or a salt thereof; or a combination thereof The composition of claim 1 further comprising:

3. LG is R ＬＧ -L ＬＧ and R ＬＧ teeth, 【Chemistry 1】 R ｃ -(Xaa)z-, a nucleic acid moiety, or a small molecule moiety; each Xaa is independently a residue of an amino acid or amino acid analog; t is 0 to 50; z is 1 to 50; Each R ｃ are independently -L ａ -R', Each L ａ are independently a covalent bond or C １ ~C ２０ Aliphatic or C having 1 to 5 heteroatoms １ ~C ２０ an optionally substituted divalent radical selected from heteroaliphatic, wherein one or more methylene units of said radical are independently selected from C(R') ２ -, -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O) ２ -, -S(O) ２ may be replaced by -N(R')-, -C(O)S-, or -C(O)O-; Each -Cy- is independently an optionally substituted divalent monocyclic, bicyclic, or polycyclic group, and each monocyclic ring is independently ３～２０ Alicyclic ring, C ６～２０ an aryl ring, a 5-20 membered heteroaryl ring having 1-10 heteroatoms, and a 3-20 membered heterocyclyl ring having 1-10 heteroatoms; L ＬＧ Is, -L ＬＧ１ -, -L ＬＧ１ -L ＬＧ２ -, -L ＬＧ１ -L ＬＧ２ -L ＬＧ３ - or -L ＬＧ１ -L ＬＧ２ -L ＬＧ３ -L ＬＧ４ - and RG is -L ＲＧ１ -L ＲＧ２ -, -L ＬＧ４ -L ＲＧ１ -L ＲＧ２ -, -L ＬＧ３ -L ＬＧ４ -L ＲＧ１ -L ＲＧ２ - or -L ＬＧ２ -L ＬＧ３ -L ＬＧ４ -L ＲＧ１ -L ＲＧ２ - and L ＬＧ１ , L ＬＧ２ , L ＬＧ３ , L ＬＧ４ , L ＲＧ１ , L ＲＧ２ , and L ＲＭ each is independently L; Each L is independently a covalent bond or a divalent, optionally substituted, straight chain or branched C alkyl group containing one or more aliphatic moieties, aryl moieties, heteroaliphatic moieties each independently having 1 to 20 heteroatoms, heteroaromatic moieties each independently having 1 to 20 heteroatoms, or any combination of any one or more of such moieties. １～１００ one or more methylene units of said group are independently １～６ Alkylene, C １～６ Alkenylene, divalent C having 1 to 5 heteroatoms １～６ Heteroaliphatic groups, -C≡C-, -Cy-, -C(R') ２ -, -O-, -S-, -S-S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -C(O)C(R') ２ N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O) ２ -, -S(O) ２ N(R')-, -C(O)S-, -C(O)O-, -P(O)(OR')-, -P(O)(SR')-, -P(O)(R')-, -P(O)(NR')-, -P(S)(OR')-, -P(S )(SR')-, -P(S)(R')-, -P(S)(NR')-, -P(R')-, -P(OR')-, -P(SR')-, -P(NR')-, amino acid residue, or -[(-OC(R') ２ -C(R') ２ -) ｎ ]- (wherein n is 1 to 20), Each R' is independently -R, -C(O)R, -CO ２ R or -SO ２ R, Each R is independently -H or C １～３０ Aliphatic, C with 1-10 heteroatoms １～３０ Heteroaliphatic, C ６～３０ Aryl, C ６～３０ Arylaliphatic, C with 1-10 heteroatoms ６～３０ an optionally substituted group selected from arylheteroaliphatic, 5-30 membered heteroaryl having 1-10 heteroatoms, and 3-30 membered heterocyclyl having 1-10 heteroatoms; or The two R groups may independently be taken together to form a covalent bond, or two or more R groups on the same atom may independently, together with said atom, form an optionally substituted 3-30 membered monocyclic, bicyclic, or polycyclic ring having, in addition to said atom, 0-10 heteroatoms; or two or more R groups on two or more atoms may independently, together with their intervening atoms, form an optionally substituted 3-30 membered monocyclic, bicyclic, or polycyclic ring having, in addition to said intervening atoms, 0-10 heteroatoms; The composition according to claim 1 or 2.

4. The composition of claim 1 or 2, wherein LG is or comprises a target binding moiety that binds to a targeting agent, said targeting agent being an antibody agent.

5. LG is or comprises a target binding moiety that binds to the Fc region, and / or ＬＧ The composition of claim 3, wherein the amino acid sequence is or comprises DCAWXLGELVWCT (SEQ ID NO: 1), wherein the two cysteine ​​residues may form a disulfide bond, and X is an amino acid residue.

6. The following conditions (a) the moiety of interest is or comprises a therapeutic agent; (b) the moiety of interest is or comprises a moiety capable of binding to a protein, a nucleic acid, or a cell; and / or (c) the moiety of interest is or comprises a reactive moiety suitable for a bioorthogonal reaction; The composition of claim 3, wherein at least one of the following is satisfied.

7. LG is a compound represented by the formulas A-1 to A-50: 【Chemistry 2】 【Chemistry 3】 【Chemistry 4】 【Chemistry 5】 【Chemistry 6】 【Chemistry 7】 【Chemistry 8】 【Chemistry 9】 【Chemistry 10】 【Chemistry 11】 【Chemistry 12】 【Chemistry 13】 3. The composition of claim 1 or 2, which is or comprises a target binding moiety having a structure selected from:

8. 3. The composition of claim 1 or 2, wherein the MOI is or comprises a therapeutic drug moiety and / or the MOI is or comprises an antibody drug.

9. L ＬＭ is one or more -[(CH ２ ) ｎ -O] ｍ -, wherein each n is independently 1 to 20 and m is 1 to 100.

10. L ＲＭ The linker may be one or more -[(CH ２ ) ｎ -O] ｍ -, wherein each n is independently 1 to 20 and m is 1 to 100.

11. RG is a group of the formula -L ＬＧ２ , -L ＬＧ２ -L ＬＧ３ -L ＬＧ４ -L ＲＧ１ - or -L ＲＧ１ -L ＲＧ２ - group, wherein L ＬＧ２ is -NH-, -NHCO-, -(CH ２ ) ｎ -NHCO-, -(CH ２ ) ｎ -OCO-, -(CH ２ ) ｎ -OCONH-, -C(O)-NHCH ２ -, -C(O)-NHCH ２ CH ２ -, -C(O)O-CH ２ (-), or -NH-C(O)O-CH ２ -, and L ＬＧ３ is an optionally substituted aryl ring, L ＬＧ４－ is a bond, —NH— or —O—, L ＲＧ１ -O-C(O)-, -C(O)-, -S(O)-, -OS(O) ２ -, or -OP(O(OR)-, L ＲＧ２ is -CH ２ -C(O)-, -C(O)-, or -CH ２ - and L ＬＧ is -(O)C-[(CH ２ ) ｎ O] ｍ (CH ２ ) ｎ NH-, -(O)C-[(CH ２ ) ｎ O] ｍ (CH ２ ) ｎ NH-, -[(CH ２ ) ｎ O] ｍ NH C(O)[(CH ２ ) ｎ O] ｍ NH-, -[(CH ２ ) ｎ O] ｍ {NH C(O)[(CH ２ ) ｎ O] ｍ}{{END]] ｐ NH-, -[(CH ２ ) ｎ O] ｍ Cy[(CH ２ ) ｎ O] ｍ NH-, -[(CH ２ ) ｎ O] ｍ Cy[(CH ２ ) ｎ O] ｍ NH C(O)[(CH ２ ) ｎ O] ｍ NH-, or -[(CH ２ ) ｎ O] ｍ Cy[(CH ２ ) ｎ O] ｍ {NH C(O)[(CH ２ ) ｎ O] ｍ}{{END]] ｐ NH- and wherein n, m, and p at each occurrence are integers independently selected from 1 to 12, and Cy is an optionally substituted cyclic group. The composition according to claim 1 or 2.

12. RG is a group represented by the formula -L ＬＧ２ -L ＬＧ３ -L ＬＧ４ -L ＲＧ１ Based on 【Chemistry 14】 The composition of claim 11 , wherein the compound is selected from the group consisting of

13. 12. The composition of claim 11, wherein n is 2 at each occurrence.

14. The composition of claim 11, wherein the reactive group is or includes -C(O)-O- or -O-C(O)-.

15. 12. The composition of claim 11, wherein the reactive group comprises an aryl group, optionally attached to -C(O)-O- or -O-C(O)-, and substituted with one or more electron-withdrawing groups.

16. The aryl group is 【Chemistry 15】 wherein R ｓ represents independently at each occurrence a halogen, -NO ２ , -F, -LR', -C(O)-LR', -S(O)-LR', -S(O) ２ -L-R', and -P(O)(-L-R') ２ and R' is selected from H or C １ ~C ６ The composition of claim 11 wherein the alkyl group is an alkyl group.

17. 3. The composition of claim 2, wherein the first compound or salt thereof and the second compound or salt thereof are present in equimolar amounts.

18. In formula (R-I), the targeting agent is an antibody comprising an IgG heavy chain comprising K246 or K248; The target binding moiety brings the reactive group into close proximity with K246 or K248 of the IgG heavy chain to allow a reaction between K246 or K248 and the reactive group, and ＲＭ - configured to bind to said antibody in a manner that results in attachment of a moiety comprising an MOI and removal of a group containing a target binding moiety from said compound or a salt thereof; The composition of claim 2.

19. 1. A composition comprising: Formula (P-II): P-N-L ＰＭ (a) [During the ceremony, P-N is a protein drug moiety containing a lysine residue; L ＰＭ is the linker, MOI is the moiety of interest. or a salt thereof, structure: LG-OH (LG-I) where LG is a group that contains a target binding moiety that binds to a targeting agent. and salts thereof, Formula (RI): LG-RG-L ＲＭ (a) [In the formula, LG is a group containing a target binding moiety that binds to a targeting drug, and is the same as LG in formula (LG-I); RG is a reactive group, L ＲＭ is a linker and is the same as L RM in formula (P-II), MOI is the moiety of interest. and salts thereof. and at least one of A composition comprising:

20. Formula (R-III): HO-RG-L ＲＭ -MOI(R---I) or a salt thereof, or a combination thereof.

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