Trans-cyclooctene modified target protein degradation inducer conjugate

Abstract

This disclosure generally relates to trans-cyclooctene modified TPD conjugates comprising a target protein degradation inducer (TPD) covalently bound to at least one trans-cyclooctene moiety via an optional linker, the conjugate having applications, for example, in the treatment of cancer, tumor growth, and immunotherapy.

Description

[Technical Field] 【0001】 Cross-reference of related applications This application claims the benefit of U.S. Provisional Application No. 63 / 503,126, filed on 18 May 2023 under 35 U.S. Code § 119(e), which is incorporated herein by reference in its entirety. 【0002】 This disclosure generally relates to trans-cyclooctane modified targeted proteolytic (TPD) conjugates comprising a targeted proteolytic (TPD) covalently bound to at least one trans-cyclooctane moiety via an optional linker, the conjugate having applications, for example, in the treatment of cancer, tumor growth, and immunotherapy. [Background technology] 【0003】 Targeted proteolytic inducers (TPDs), sometimes referred to as proteolytic chimeras (PROTACs), are engineered techniques for targeted proteolysis and are emerging as a promising approach for targeted therapy in various diseases, particularly cancer. Bifunctional TPD molecules typically consist of two covalent ligands that recruit a target protein and an E3 ubiquitin ligase together to trigger proteasomal degradation of the target protein by the ubiquitin-proteasome system (UPS). 【0004】 For proper function, TPDs need to enter cells to recruit intracellular UPSs, therefore their membrane permeability is key to TPD function. However, the large molecular weight and large exposed polar surface area of ​​TPDs can result in low cell / tissue permeability. Furthermore, some E3 ligands exhibit low specificity, leading to such TPDs having off-target effects. 【0005】 Therefore, there is a need to provide modified TPDs for the effective and safe delivery of TPDs in vivo. [Overview of the Initiative] 【0006】 Bioorthogonal conjugations, or click reactions, are selective and orthogonal (non-interacting) functions found in biological systems that have found use in a variety of applications in the fields of chemistry, chemical biology, molecular diagnostics, and medicine. They can be used to facilitate the selective manipulation of molecules, cells, particles, and surfaces, as well as the tagging and tracking of biomolecules in vitro and in vivo. 【0007】 Provided herein are trans-cyclooctene modified target proteolytic inducer (TPD) (or PROTAC) conjugates comprising a target proteolytic inducer (TPD) covalently bound to at least one trans-cyclooctene moiety via an optional linker. The trans-cyclooctene modified TPD conjugates described herein are designed to be used with a tetrazine activator, which then releases the TPD in vivo upon contact with the trans-cyclooctene modified TPD conjugate. 【0008】 The trans-cyclooctene modified TPD conjugates described herein, but not limited to these, are intended to have enhanced advantages, such as improving TPD permeability or masking TPD activity to improve the safety profile. It is further intended that the cellular permeability of TPD can be increased by using a linker between the TPD and the trans-cyclooctene moiety, or by derivatizing one or more trans-cyclooctene moieties to form intramolecular hydrogen bonds that partially reduce polarity, or by conjugating a cellular permeable peptide, such as a poly-D-arginine sequence, to it. 【0009】 In a particular embodiment, the conjugates shown in Table 1 are provided. 【0010】 In some embodiments, a method for treating cancer is provided, the method comprising administering a conjugate described herein to a subject in need thereof, and administering a tetrazine activator described herein to the subject. 【0011】 In addition, compounds comprising TPD precursors that react individually upon administration to produce TPD in vivo are described herein. Thus, in certain embodiments, a system comprising a trans-cyclooctene-modified E3 ubiquitin ligase ligand and a tetrazine-modified target protein ligand is provided. A system comprising a tetrazine-modified E3 ubiquitin ligase ligand and a trans-cyclooctene-modified target protein ligand is also provided. 【0012】 In certain embodiments, methods are provided for treating cancer or enhancing or inducing an immune response, the methods comprising administering to a subject in need thereof a therapeutically effective amount of a trans-cyclooctene-modified E3 ubiquitin ligase ligand and a tetrazine-modified target protein ligand, or a tetrazine-modified E3 ubiquitin ligase ligand and a trans-cyclooctene-modified target protein ligand. 【0013】 In a particular embodiment, the compounds listed in Table 2 are provided. In a particular embodiment, a kit containing two or more compounds from Table 2 is provided. 【0014】 In some embodiments, the cancer is metastatic. In some embodiments, the cancer is melanoma, kidney cancer, prostate cancer, ovarian cancer, endometrial cancer, breast cancer, glioblastoma, lung cancer, soft tissue sarcoma, fibrosarcoma, osteosarcoma, pancreatic cancer, gastric cancer, squamous cell carcinoma of the head and neck, anal cancer / vulvar cancer, esophageal cancer, pancreatic adenocarcinoma, cervical cancer, hepatocellular carcinoma, Kaposi's sarcoma, non-Hodgkin lymphoma, Hodgkin lymphoma, Wilms' tumor / neuroblastoma, bladder cancer, thyroid adenocarcinoma, pancreatic neuroendocrine tumor, prostate adenocarcinoma, nasopharyngeal cancer, or cutaneous T-cell lymphoma. 【0015】 In some embodiments, cancer is melanoma, kidney cancer, prostate cancer, ovarian cancer, breast cancer, glioma, lung cancer, soft tissue cancer, soft tissue sarcoma, osteosarcoma, or pancreatic cancer. In some embodiments, cancer is a solid tumor. In some embodiments, cancer is lymphoma or leukemia. In some embodiments, cancer is a hematological malignancy. [Modes for carrying out the invention] 【0016】 The following description illustrates exemplary embodiments of the Technology. However, such descriptions should be understood not as limiting the scope of the Disclosure, but rather as being provided solely for illustrative purposes. 【0017】 1.Definition For clarity, certain features of the Disclosure described in the context of separate embodiments may also be provided in combination in a single embodiment. Conversely, for brevity, various features of the Disclosure described in the context of a single embodiment may also be provided separately or in any preferred partial combination. All combinations of embodiments belonging to the Disclosure are specifically encompassed by the Disclosure and are disclosed herein as if each and all combinations were individually and explicitly disclosed to the extent that such combinations encompass subject matter that is, for example, a stable compound (i.e., a compound that can be prepared, isolated, characterized, and further tested for biological activity). Furthermore, all partial combinations of various embodiments and their elements (e.g., elements of chemical groups enumerated in embodiments describing such variables) are also specifically encompassed by the Disclosure and are disclosed herein as if each and all such partial combinations were individually and explicitly disclosed herein. 【0018】 A.Definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art. In case of any conflict, this specification, including its definitions, shall prevail. Preferred methods and materials are described below, but similar or equivalent methods and materials may be used in the practice or testing of this disclosure. All publications, patent applications, patents, and other references referenced herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting. 【0019】 The terms “comprise(s),” “include(s),” “have,” “possess,” “can have,” and “contain,” and their variations, when used herein, are intended to be open-ended transitional phrases, terms, or words that do not preclude the possibility of additional acts or structures. The singular forms “a,” “an,” and “the” include multiple references unless the context otherwise explicitly indicates. This disclosure also contemplates other embodiments that “include,” “consist of,” and “essentially consist of,” the embodiments or elements presented herein, whether expressly described or not. 【0020】 The modifier "approximately" used in relation to quantity includes the stated value and has a meaning determined by the context (for example, at least the degree of error associated with measuring a particular quantity). The modifier "approximately" should also be considered to disclose a range defined by the absolute values ​​of two endpoints. For example, the expression "approximately 2 to approximately 4" also discloses a range of "2 to 4". The term "approximately" can refer to plus or minus 10% of the stated number. For example, "approximately 10%" may indicate a range of 9% to 11%, and "approximately 1" may mean 0.9 to 1.1. Other meanings of "approximately" may become apparent from contexts such as rounding, so for example, "approximately 1" may also mean 0.5 to 1.4. 【0021】 The conjunction “or” includes any and all combinations of the one or more enumerated elements associated by the conjunction. For example, the phrase “a device containing A or B” could mean a device containing A without B, a device containing B without A, or a device containing both A and B. The phrases “at least one of A, B, ... and N” or “at least one of A, B, ... N, or any combination thereof” are defined in the broadest sense to mean one or more elements selected from the group containing A, B, and N, i.e., any combination of one or more elements A, B, ... or N that includes any one element alone or in combination with one or more of the other elements, and may also include additional elements not enumerated. 【0022】 The definitions of specific functional groups and chemical terms are described in more detail below. For the purposes of this disclosure, chemical elements are defined as follows: Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75 th Identified according to the ed. and inside cover, specific functional groups are generally defined as described therein. In addition, general principles of organic chemistry, as well as specific functional parts and reactivity, are referred to in *Organic Chemistry*, Thomas Sorrell, University Science Books, Sausalito, 1999, and *Smith and March March's Advanced Organic Chemistry*, 5 th Edition, John Wiley&Sons, Inc., New York, 2001, Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989, Carruthers, Some Modern Methods of Organic Synthesis, 3 rdThis information is contained in Edition, Cambridge University Press, Cambridge, 1987, and the entirety of each of its contents is incorporated herein by reference. 【0023】 As used herein, the term "alkyl" means a straight or branched saturated hydrocarbon chain containing 1 to 30 carbon atoms. 1- The term "C6-alkyl" refers to a linear or branched hydrocarbon containing 1 to 6 carbon atoms. The term "C1-C3-alkyl" refers to a linear or branched hydrocarbon containing 1 to 3 carbon atoms. Typical examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, and n-decyl. 【0024】 As used herein, the term "alkoxy" refers to an alkyl group as defined herein, which is attached to the parent molecule via an oxygen atom. Typical examples of alkoxys include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, and tert-butoxy. 【0025】 As used herein, the term "alkenyl" means a hydrocarbon chain containing 2 to 30 carbon atoms having at least one carbon-carbon double bond. The alkenyl group may be substituted or unsubstituted. For example, the alkenyl group may be substituted with an aryl group such as phenyl. 【0026】 As used herein, the term "alkynyl" refers to a linear or branched monovalent hydrocarbyl group having 2 to 30 carbon atoms, e.g., 2 to 20 or 2 to 10 carbon atoms, and having at least one triple-unsaturated site. The term "alkyne" also includes non-aromatic cycloalkyl groups having 5 to 20 carbon atoms, e.g., 5 to 10 carbon atoms, and having one or more rings and at least one triple bond. Examples of such alkynyl groups include, but are not limited to, acetylenyl (-C≡CH) and propargyl (-CH2C≡CH), and cycloalkynyl moieties such as substituted or unsubstituted cyclooctin moieties. 【0027】 When used herein, the term "alkoxyalkyl" refers to an alkoxy group as defined herein, attached to the parent molecule via an alkyl group as defined herein. 【0028】 As used herein, the term "alkylene" refers to a divalent group derived from a linear or branched hydrocarbon having 1 to 30 carbon atoms, for example, 2 to 10 carbon atoms. Typical examples of alkylenes include, but are not limited to, -CH2-, -CH(CH3)-, -C(CH3)2-, -CH2CH2-, -CH(CH3)CH2-, -C(CH3)2CH2-, -CH2CH2CH2-, -CH(CH3)CH2CH2-, -C(CH3)2CH2CH2-, -CH2C(CH3)2CH2-, -CH2CH2CH2CH2-, and -CH2CH2CH2CH2CH2-. 【0029】 The term "amino acid" refers to both natural and unnatural amino acids, protected natural and unnatural amino acids, and amino acid analogs and mimics that function in a similar manner to naturally occurring amino acids. Naturally encoded amino acids include 20 common amino acids (alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine), as well as pyrrolidine and selenocysteine. Unnatural amino acids refer to amino acid analogs that have the same basic chemical structure as naturally occurring amino acids, i.e., α-carbons bonded to hydrogen, carboxyl groups, amino groups, and R groups, for example only. Such analogs may have modified R groups (e.g., norleucine, for example) or may retain a modified peptide skeleton while retaining the same basic chemical structure as natural amino acids. Non-exclusive examples of unnatural amino acids or amino acid analogs include citrulline, homoserine, norleucine, methionine sulfoxide, methionine methylsulfonium, homophenylalanine, ornithine, formylglycine, phenylglycine, para-azidophenylglycine, para-azidophenylalanine, para-acetophenylalanine, 4-(3-methyl-(1,2,4,5-tetrazine))-phenylglycine, and 4-(3-methyl-(1,2,4,5-tetrazine))-phenylalanine. 【0030】 The term "aryl," as used herein, refers to an aromatic carbocyclic group having a single ring (e.g., monocyclic) or multiple rings including a condensed system (e.g., bicyclic or tricyclic). Typical examples of aryls include, but are not limited to, phenyl, naphthyl, and anthracenyl. Monocyclic, bicyclic, and tricyclic aryls are connected to the parent molecule via any carbon atoms contained within the ring and can be unsubstituted or substituted. Aromatic bicyclic or aromatic tricyclic ring systems do not contain a non-aromatic ring. Therefore, if a bicyclic or tricyclic ring system contains a non-aromatic ring, the ring system is cycloalkyl or heterocyclyl, depending on whether the heteroatom is present on the non-aromatic ring, regardless of the bonding site to the rest of the molecule. 【0031】 In some embodiments, the term “aryl” as used herein refers to a phenyl group, or a bicyclic or tricyclic aryl fused ring system. A bicyclic fused ring system is exemplified by a phenyl group attached to the parent molecule and fused to a phenyl group. A tricyclic fused ring system is exemplified by a phenyl group attached to the parent molecule and fused to two other phenyl groups. Typical examples of bicyclic aryls include, but are not limited to, naphthyl. Typical examples of tricyclic aryls include, but are not limited to, anthracenyl. Monocyclic, bicyclic, and tricyclic aryls can be attached to the parent molecule via any carbon atoms contained within the ring and can be unsubstituted or substituted. 【0032】 As used herein, the term "azide" refers to the functional group -N3. 【0033】 As used herein, the term "cycloalkyl" refers to a non-aromatic carbocyclic ring system containing 3 to 10, or 3 to 8, or 3 to 6, or 5 to 10 carbon atoms and zero heteroatoms. A cycloalkyl ring system may contain one or more double bonds, provided the ring is not aromatic; therefore, the term cycloalkyl includes cycloalkenyl ring systems. Typical examples of cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, and cyclodecyl. Exemplary monocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl, or cycloheptenyl. "Cycloalkyl" also includes carbocyclic ring systems in which the cycloalkyl group is condensed to an aryl or heteroaryl as defined herein, regardless of its bonding site to the rest of the molecule. 【0034】 In some embodiments, the term “cycloalkyl” refers, as used herein, to a carbocyclic ring system containing 3 to 10 carbon atoms, zero heteroatoms, and zero double bonds. Typical examples of cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, and cyclodecyl. “Cycloalkyl” also includes carbocyclic ring systems in which a cycloalkyl group is added to a parent molecule and condensed into an aryl group, a heteroaryl group, or a heterocycle as defined herein. 【0035】 As used herein, the term "cycloalkenyl" means a non-aromatic monocyclic or polycyclic ring system containing at least one carbon-carbon double bond and preferably having 5 to 10 carbon atoms per ring. Examples of monocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl, or cycloheptenyl. 【0036】 As used herein, the term "cyclooctene" refers to a substituted or unsubstituted non-aromatic cyclic alkyl group comprising eight carbon atoms and having a single ring with a double bond. Examples of such cyclooctene groups include, but are not limited to, substituted or unsubstituted trans-cyclooctene (TCO). 【0037】 As used herein, the term "fluoroalkyl" means an alkyl group as defined herein, in which 1, 2, 3, 4, 5, 6, 7, or 8 hydrogen atoms are replaced by fluorine. Typical examples of fluoroalkyls include, but are not limited to, 2-fluoroethyl, 2,2,2-trifluoroethyl, trifluoromethyl, difluoromethyl, pentafluoroethyl, and trifluoropropyl, for example, 3,3,3-trifluoropropyl. 【0038】 When used herein, the term "alkoxyfluoroalkyl" refers to an alkoxy group as defined herein, which is attached to the parent molecule via a fluoroalkyl group, as defined herein. 【0039】 As used herein, the term "fluoroalkoxy" means at least one fluoroalkyl group as defined herein, attached to the parent molecule via an oxygen atom. Typical examples of fluoroalkyloxys include, but are not limited to, difluoromethoxy, trifluoromethoxy, and 2,2,2-trifluoroethoxy. 【0040】 The terms "halogen" or "halo" as used herein mean Cl, Br, I, or F. 【0041】 As used herein, the term "haloalkyl" means an alkyl group as defined herein, in which one, two, three, four, five, six, seven, or eight hydrogen atoms are replaced by halogens. 【0042】 When used herein, the term "haloalkoxy" means at least one haloalkyl group as defined herein, attached to the parent molecule via an oxygen atom. 【0043】 As used herein, the term "heteroalkyl" means an alkyl group as defined herein, in which one or more carbon atoms are replaced by heteroatoms selected from S, Si, O, P, and N. Heteroatoms can be oxidized. Typical examples of heteroalkyls include, but are not limited to, alkyl ethers, secondary and tertiary alkylamines, and alkyl sulfides. 【0044】 As used herein, the term "heteroaryl" refers to an aromatic group having a monocyclic, polycyclic, or multiple fused ring, where one or more ring heteroatoms are independently selected from nitrogen, oxygen, and sulfur. In some embodiments, as used herein, the term "heteroaryl" refers to an aromatic monocyclic ring, an aromatic bicyclic ring system, or an aromatic tricyclic ring system. An aromatic monocyclic ring is a 5- or 6-membered ring containing at least one heteroatom independently selected from the group consisting of N, O, and S (e.g., 1, 2, 3, or 4 heteroatoms independently selected from O, S, and N). A 5-membered aromatic monocyclic ring has two double bonds, and a 6-membered aromatic monocyclic ring has three double bonds. Representative examples of monocyclic heteroaryls include, but are not limited to, pyridinyl (including pyridine-2-yl, pyridine-3-yl, and pyridine-4-yl), pyrimidinyl, pyrazinyl, thienyl, furyl, thiazolyl, thiadiazolyl, isoxazolyl, pyrazolyl, and 2-oxo-1,2-dihydropyridinyl. Representative examples of bicyclic heteroaryls include, but are not limited to, clomenyl, benzothienyl, benzodioxolyl, benzotriazolyl, quinolinyl, thienopyrrolyl, thienothienyl, imidazothiazolyl, benzothiazolyl, benzofuranil, indolyl, quinolinyl, imidazopyridine, benzoxadiazolyl, and benzopyrazolyl. Representative examples of tricyclic heteroaryls include, but are not limited to, dibenzofuranil and dibenzothienyl. Monocyclic, bicyclic, and tricyclic heteroaryls are linked to the parent molecule via any carbon or nitrogen atom contained within the ring and can be unsubstituted or substituted. In some embodiments, aromatic bicyclic or aromatic tricyclic ring systems do not contain a non-aromatic ring. Therefore, if a bicyclic or tricyclic ring system contains a non-aromatic ring, the ring system is cycloalkyl or heterocyclyl, depending on whether the heteroatom is present in the non-aromatic ring, regardless of the bonding site to the rest of the molecule. 【0045】 In some embodiments, a five-membered aromatic monocyclic ring has two double bonds, and a six-membered aromatic monocyclic ring has three double bonds. In some embodiments, exemplary bicyclic heteroaryl groups are exemplified by monocyclic heteroaryl rings fused to a parent molecule and a monocyclic cycloalkyl group, a monocyclic aryl group, a monocyclic heteroaryl group, or a monocyclic heterocycle as defined herein. In some embodiments, tricyclic heteroaryl groups are exemplified by monocyclic heteroaryl rings fused to two of a parent molecule and a monocyclic cycloalkyl group, a monocyclic aryl group, a monocyclic heteroaryl group, or a monocyclic heterocycle as defined herein. 【0046】 As used herein, the terms “heterocyclyl,” “heterocycle,” or “heterocyclic” refer to a non-aromatic ring system containing 3 to 10, or 3 to 8, or 3 to 6, or 5 to 10 carbon atoms, at least one heteroatom (e.g., 1 to 5, 1 to 4, 1 to 3, 1 to 2, or 1), and optionally one or more oxo and / or double bonds. The terms “heterocyclyl,” “heterocycle,” or “heterocyclic” include monocyclic, bicyclic, tricyclic, fused, spirocyclic, or bridging ring systems, provided that at least one non-aromatic ring system containing at least one heteroatom is present. In some embodiments, a monocyclic heterocycle is a 3, 4, 5, 6, 7, or 8-membered ring containing at least one heteroatom independently selected from the group consisting of O, N, and S. In some embodiments, the 3- or 4-membered ring contains zero or one double bond and one heteroatom selected from the group consisting of O, N, and S. In some embodiments, the 5-membered ring contains zero or one double bond and one, two, or three heteroatoms selected from the group consisting of O, N, and S. In some embodiments, the 6-membered ring contains zero, one, or two double bonds and one, two, or three heteroatoms selected from the group consisting of O, N, and S. In some embodiments, the 7- and 8-membered rings contain zero, one, two, or three double bonds and one, two, or three heteroatoms selected from the group consisting of O, N, and S.Typical examples of monocyclic heterocycles include azetidinyl, azepanyl, azilidinyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, 1,3-dithiolanyl, 1,3-dithianyl, 1,3-dimethylpyrimidine-2,4(1H,3H)-dione, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolinyl, isoxazolinyl, isoxazolinyl, morpholinyl, oxadiazolinyl, oxadiazolidinyl, oxazolinyl, oxazolidinyl, and oxetanyl. Examples include, but are not limited to, piperazinil, piperidinil, pyranil, pyrazolinil, pyrazolidinil, pyrrolinil, pyrrolidinil, tetrahydrofuranil, tetrahydropyranil, tetrahydropyridinil, tetrahydrothienyl, thiadiazolinil, thiadiazolidinil, 1,2-thiadinil, 1,3-thiadinil, thiazolinil, thiazolidinil, thiomorpholinil, 1,1-dioxidethiomorpholinil (thiomorpholine sulfone), thiopyranil, and trithianil. A bicyclic heterocycle is a monocyclic heterocycle fused to a phenyl group, or a monocyclic heterocycle fused to a monocyclic cycloalkyl group, or a monocyclic heterocycle fused to a monocyclic cycloalkenyl group, or a monocyclic heterocycle fused to a monocyclic heterocycle, or a spiroheterocyclic group, or a bridged monocyclic heterocycle system in which two non-adjacent atoms of the ring are linked by an alkylene bridge of 1, 2, 3, or 4 carbon atoms, or an alkenylene bridge of 2, 3, or 4 carbon atoms. Representative examples of bicyclic heterocycles include, but are not limited to, benzopyranyl, benzothiopyranyl, chromanyl, 2,3-dihydrobenzofuranyl, 2,3-dihydrobenzothienyl, 2,3-dihydroisoquinoline, 2-azaspiro[3.3]heptan-2-yl, azabicyclo[2.2.1]heptyl (including 2-azabicyclo[2.2.1]hepta-2-yl), 2,3-dihydro-1H-indolyl, isoindolinyl, octahydrocyclopenta[c]pyrrolyl, octahydropyrrolopyridinyl, and tetrahydroisoquinolinyl.Tricyclic heterocycles are exemplified by bicyclic heterocycles fused to a phenyl group, or bicyclic heterocycles fused to a monocyclic cycloalkyl group, or bicyclic heterocycles fused to a monocyclic cycloalkenyl group, or bicyclic heterocycles fused to a monocyclic heterocycle, or bicyclic heterocycles in which two non-adjacent atoms of a bicyclic ring are linked by an alkylene bridge of 1, 2, 3, or 4 carbon atoms, or an alkenylene bridge of 2, 3, or 4 carbon atoms. Examples of tricyclic heterocycles include octahydro-2,5-epoxypentalene, hexahydro-2H-2,5-methanocyclopenta[b]furan, hexahydro-1H-1,4-methanocyclopenta[c]furan, and aza-adamantane(1-azatricyclo[3.3.1.1). 3,7 ]decane), and oxa-adamantane (2-oxatricyclo[3.3.1.1 3,7 Examples include, but are not limited to, decanes. Monocyclic, bicyclic, and tricyclic heterocyclic rings are connected to the parent molecule via any carbon or nitrogen atoms contained within the ring and can be unsubstituted or substituted. 【0047】 As used herein, the term "hydroxyl" means the -OH group. 【0048】 As used herein, the term "hydroxyalkyl" means an alkyl group as defined herein, in which one, two, three, four, five, six, seven, or eight hydrogen atoms are replaced by a hydroxyl group. 【0049】 The term "substituted" refers to a group that can be further substituted with one or more nonhydrogen substituent groups. Examples of substituent groups include, but are not limited to, halogens, =O, =S, cyano, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclic, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkylene, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, -COOH, ketones, amides, carbamates, and acyls. 【0050】 The term "tetrazine" refers to a substituted or unsubstituted aromatic cyclic group having two carbon atoms and four nitrogen atoms, with a single ring having three double bonds. Examples of tetrazine groups include 1,2,3,4-tetrazine and 1,2,4,5-tetrazine. As used herein, 1,2,4,5-tetrazine is referred to as the "Tz" group. The term "tetrazine-based" generally refers to a moiety that can be converted to tetrazine as defined herein, or to tetrazine such as optionally substituted dihydrotetrazine. 【0051】 The term “selective delivery” refers to the delivery of a drug (e.g., TPD) to an organ or tissue (or part thereof) requiring treatment or diagnosis without significant binding to other non-target organs or tissues (or parts thereof). In some embodiments, the trans-cyclooctene modified TPD conjugates described herein do not have therapeutic effects themselves, but rather are designed to enable the selective or targeted delivery of TPD. However, tetrazine-based targeting agents may have therapeutic effects, and therefore such constructs are not excluded by this disclosure. 【0052】 The term "diagnostic agent" refers to an agent that aids in the diagnosis of a condition or disease. Representative diagnostic agents include imaging agents such as paramagnetic agents, optical probes, and radionuclides. A paramagnetic agent is an imaging agent that is magnetic under an externally applied magnetic field. Examples of paramagnetic agents include, but are not limited to, iron particles including iron nanoparticles and iron microparticles. An optical probe is a fluorescent compound that can be detected by excitation with radiation of one wavelength and detection with radiation of a different second wavelength. Optical probes of the present disclosure include, but are not limited to, Cy5.5, Alexa 680, Cy5, DiD (1,1'-dioctadecyl-3,3,3',3'-tetramethylindodicarbocyanine perchlorate), and DiR (1,1'-dioctadecyl-3,3,3',3'-tetramethylindotricarbocyanine iodide). Other optical probes include quantum dots. A radionuclide is an element that undergoes detectable radioactive decay. Radionuclides useful in embodiments of the present disclosure include 3 H, 11 C, 13 N, 18 F, 19 F, 60 Co, 64 Cu, 67 Cu, 68 Ga, 82 Rb, 89 Zr, 90 Sr, 90 Y, 99 Tc, 99m Tc, 111 In, 123 I, 124 I, 125 I, 129 I, 131 I, 137 Cs, 177 Lu, 186 Re, 188 Re, 211 At, Rn, Ra, Th, U, Pu, and 241 Am, but are not limited to these. 【0053】 The term “ligand” as used herein is used to define a binding partner for a biological target molecule (e.g., an enzyme or receptor) in vivo. Ligands for use according to the present invention are typically small molecules that can be functionalized, optionally, via a linker by the addition of a TCO moiety. 【0054】 The term “targeting agent” refers to a chemical or biological agent that specifically binds to a target (e.g., a targeted organ or tissue) and thereby forms a stable association between the targeting agent and the specific target. “Stable association” means that a compound associates with another part or structure, for example, under standard physiological conditions, either covalently or noncovalently. Stable associations, or bindings, may include covalent and noncovalent interactions, but are not limited to, ionic bonds, hydrophobic interactions, hydrogen bonds, van der Waals forces (e.g., London dispersion forces), and dipole-dipole interactions. Examples of targeting agents include ligands that specifically (or substantially specifically) bind to a particular clinically relevant target receptor or cell surface target. Specific targeting agents are described herein. 【0055】 The term "targeted organ or tissue" refers to an organ or tissue that is targeted for TPD delivery. Typical organs and tissues for targeting include those that can be targeted by chemical or biological targeting agents, as well as those that cannot be targeted by chemical or biological targeting agents. 【0056】 The term "transplantation" refers to the surgical insertion of a target into the body. 【0057】 The term "contact" or "to come into contact" refers to the process of bringing at least two different species into contact so that they can interact with each other, such as in non-covalent or covalent interactions or bonding reactions. However, it should be understood that the resulting complex or reaction product can be formed directly from interactions or reactions between added reagents, or from intermediates from one or more of the added reagents or parts that can be formed in the contact mixture. 【0058】 The term "binder" refers to a drug having a functional group capable of forming a covalent bond with a complementary functional group of another binder in a biological environment. The bonding between binders in a biological environment may also be referred to as bioconjugation. Examples of binders include bioorthogonal binders, which are binders having bioorthogonal functional groups. The bioorthogonal functional groups of a bioorthogonal binder selectively react with complementary bioorthogonal functional groups of another bioorthogonal bonding partner. Selective reactions between bioorthogonal bonding partners minimize side reactions with other binders, biological compounds, or other non-complementary bioorthogonal binders or non-complementary bioorthogonal functional groups. Examples of bioorthogonal moieties or functional groups of bioorthogonal binders include, but are not limited to, azides and alkynes, trans-cyclooctene (TCO) and tetrazine (Tz) (e.g., 1,2,4,5-tetrazine) for forming triazoles via click reactions, and others. Binders useful in this disclosure may have high reactivity with the corresponding binder so that the reaction is rapid. 【0059】 The term "functionalized" refers to a part that has a functional group attached to it, for example, a part that has a binder functional group attached to it (e.g., a bioorthoth functional group). 【0060】 The term “administering” refers to any preferred route of administration to the subject, including, but not limited to, oral administration, suppository administration, topical contact, parenteral administration, intravenous administration, intraperitoneal administration, intramuscular administration, intralesional administration, intranasal administration, or subcutaneous administration, subarachnoid administration, or implantation of a sustained-release device to the subject, such as a mini osmotic pump. 【0061】 As used herein, the term "parenteral" refers to modes of administration including intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous, and intra-articular injections and infusions. 【0062】 The term "therapeutic effective dose" refers to the amount of a compound sufficient to treat one or more of a particular disorder or disease or its symptoms, and / or to prevent or reduce the risk of the onset or recurrence of the disease or disorder or its symptoms. With respect to oncogenic proliferative disorders, the pharmacologic or therapeutic effective dose includes, among other things, an amount sufficient to shrink the tumor or reduce its growth rate. 【0063】 As used herein, the terms “subject” or “patient” include humans and mammals (e.g., mice, rats, pigs, cats, dogs, and horses). Typical subjects to which the drugs of this disclosure may be administered may include mammals, in particular primates, and especially humans. For veterinary use, suitable subjects may include, for example, livestock such as cattle, sheep, goats, cows, and pigs; poultry such as chickens, ducks, geese, and turkeys; and domesticated animals, in particular pets such as dogs and cats. For diagnostic or research use, suitable subjects may include mammals such as rodents (e.g., mice, rats, and hamsters), rabbits, primates, and pigs such as inbred pigs. 【0064】 When used herein, “to treat” or “treatment” means treating or treating a disease or medical condition or symptom(s) in a patient, such as a mammal (in particular, a human), which includes (a) improving the disease or medical condition or symptom(s) in a patient, for example, eliminating or causing regression of the disease or medical condition or symptom(s) in a patient; (b) suppressing the disease or medical condition or symptom(s) in a patient, for example, by delaying or halting the progression of the disease or medical condition or symptom(s) in a patient; or (c) reducing the symptoms of the disease or medical condition or symptom(s) in a patient. 【0065】 The term "physiological conditions" means that these conditions are suitable for living cells, including, for example, aqueous conditions such as temperature, pH, and salinity, which are primarily suitable for living cells. 【0066】 For the compounds described herein, the groups and substituents may be selected according to the allowable valencies of the atoms and substituents, and the selection and substitution result in stable compounds that do not undergo spontaneous transformations such as rearrangement, cyclization, or elimination. 【0067】 Where a numerical range is indicated, unless the context clearly indicates otherwise, each intermediate value between the upper and lower limits of that range (up to one-tenth of the lower limit unit), as well as any other values ​​or intermediate values ​​indicated within that described range, should be understood to be included in this disclosure. The upper and lower limits of these smaller ranges may be independently included within the smaller ranges and are also included in this disclosure, subject to any specifically excluded limits within the described range. If the described range includes one or both limit values, the range excluding one or both of those included limit values ​​is also included in this disclosure. 【0068】 For the purposes of enumerating numerical ranges as defined herein, each number intervening between them is explicitly intended with the same degree of precision. For example, for the range 6–9, the digits 7 and 8 are intended in addition to 6 and 9, and for the range 6.0–7.0, the digits 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are explicitly intended. 【0069】 Compounds may exist as stereoisomers containing an asymmetric or chiral center. Stereoiomers are designated "R" or "S" depending on the arrangement of substituents around the chiral carbon atom. As used herein, the terms "R" and "S" refer to the arrangements defined in IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, Pure Appl. Chem., 1976, 45:13-30. This disclosure envisions various stereoisomers and mixtures thereof, which are specifically included within the scope of this disclosure. Examples of stereoisomers include enantiomers and diastereomers, as well as mixtures of enantiomers or diastereomers. Individual stereoisomers of compounds may be prepared synthetically from commercially available starting materials containing an asymmetric or chiral center, or by the preparation of racemic mixtures followed by separation methods well known to those skilled in the art. These separation methods include (1) binding of a mixture of enantiomers to a chiral auxiliary, separation of a mixture of diastereomers obtained by recrystallization or chromatography, and optional release of optically pure products from the auxiliary (Furniss, Hannaford, Smith, and Tatchell, “Vogel’s Textbook of Practical Organic Chemistry,” 5 th As exemplified by (1989 edition, Longman Scientific & Technical, Essex CM20 2JE, England), or (2) direct separation of a mixture of optical enantiomers on a chiral chromatography column, or (3) fractional recrystallization methods. 【0070】 This disclosure also includes isotope-labeled compounds that are identical to those enumerated herein, but are not identical in that one or more atoms are replaced by atoms having atomic masses or mass numbers different from those normally found in nature. Examples of isotopes suitable for inclusion in the compounds of this disclosure include, but are not limited to, those listed above. 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S, 18 F, and 36 These include hydrogen (such as Cl), carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine. Deuterium, that is, 2 Substitution with heavier isotopes such as 1H can offer certain therapeutic benefits resulting from higher metabolic stability, e.g., increased in vivo half-life or reduced dosage requirements, and may therefore be preferable in some situations. The compound may also incorporate positron-emitting isotopes for medical imaging and positron emission tomography (PET) studies to determine receptor distribution. Suitable positron-emitting isotopes that can be incorporated are: 11 C, 13 N, 15 O, and 18 F is the isotope-labeled compound disclosed herein can generally be prepared by conventional techniques known to those skilled in the art, or by processes similar to those described in the appended examples, using appropriate isotope-labeled reagents instead of non-isotope-labeled reagents. 【0071】 B. Trans-cyclooctene modified TPD conjugate The trans-cyclooctene-modified TPD conjugates described herein are designed to localize to a target site within the subject upon administration. The trans-cyclooctene-modified TPD conjugates can be administered topically or systemically. Upon administration, a tetrazine activator can be administered, which, upon contact with the trans-cyclooctene-modified TPD conjugate in vivo, enables targeted drug delivery of TPD. 【0072】 Trans-cyclooctene modified TPD conjugates are provided herein, each comprising a target proteolytic inhibitor (TPD) covalently bonded to at least one trans-cyclooctene moiety via an optional linker. 【0073】 In some embodiments, the trans-cyclooctene modified TPD conjugate is of the formula XL-TPD, where X is the trans-cyclooctene moiety, L is the linker, and TPD is the target protein degradation inducer. 【0074】 In some embodiments of the trans-cyclooctene modified TPD conjugates disclosed herein, the TPD comprises an E3 ubiquitin ligase ligand selected from the group consisting of von Hippel-Lindou (VHL) ligand, cereblon (CRBN) ligand, mouse double micro2 homolog (MDM2) ligand, and beta-transducin repeat-containing protein (β-TrCP) ligand. 【0075】 In some embodiments of the trans-cyclooctene-modified TPD conjugates disclosed herein, the TPD comprises an E3 ubiquitin ligase ligand selected from the group consisting of thalidomide, lenalidomide, pomalidomide, MLN4924 (pebonesistat), nutrin-3, and curcumin. 【0076】 In some embodiments of the trans-cyclooctene-modified TPD conjugates disclosed herein, the TPD comprises an E3 ubiquitin ligase ligand selected from Table A, where the wavy lines indicate binding sites to the rest of the TPD. TIFF2026519211000001.tif176170 【0077】 In some embodiments of the trans-cyclooctene modified TPD conjugates disclosed herein, the TPD comprises a target protein ligand selected from the group consisting of receptor tyrosine kinase (RAF) kinase ligands (e.g., BRAF kinase ligands), steroid receptor ligands (e.g., androgen receptor (AR) ligands, estrogen receptor (ER) ligands, or progesterone receptor (PR) ligands), cyclin-dependent kinase (CDK) ligands, BTK ligands, BET ligands, and apoptosis inhibitor protein (IAP) ligands. 【0078】 In some embodiments of the trans-cyclooctene modified TPD conjugates disclosed herein, the TPD comprises a target protein ligand selected from Table B, where the wavy lines indicate binding sites to the rest of the TPD. TIFF2026519211000002.tif221170TIFF2026519211000003.tif52170 【0079】 In some embodiments of the trans-cyclooctene modified TPD conjugates disclosed herein, the TPD is selected from Table C. TIFF2026519211000004.tif205170TIFF2026519211000005.tif237170TIFF2026519211000006.tif207170 【0080】 In some embodiments of the trans-cyclooctene modified TPD conjugates disclosed herein, the trans-cyclooctene portion is of formula X, [ka] In the formula, R in each occurrence 1A is independently selected from the group consisting of C 1-4 alkyl, C 1-4 haloalkyl, and C 1-4 alkoxy; q is 0, 1, or 2; q1 is 0 or 1; R in each occurrence 1B is independently G 1 , OH, -NR 1c -C 1-4 alkylene-G 1 , -NR 1c -C 1-4 alkylene-N(R 1d )2, -NR 1c -C 1-6 alkylene-N(C 1-4 alkyl)3 + , -N(R 1c )CHR 1e CO2H, -N(R 1c )-C 1-6 alkylene-CO2H, -N(R 1f )-C 2-4 alkylene-(N(C 1-4 alkylene-CO2H)-C 2-4 alkylene) n -N(C 1-4 alkylene-CO2H)2, -N(R 1c )CHR 1e C(O)OC 1-6 alkyl, -N(R 1c )-C 1-6 alkylene-C(O)OC 1-6 alkyl, -N(R 1f )-C 2-4 alkylene-(N(C 1-4 alkylene-C(O)OC 1-6 alkyl)-C 2-4 alkylene) n -N(C 1-4 alkylene-C(O)OC 1-6 alkyl)2, -N(R 1c )-C 1-6 alkylene-SO3H, -N(R1c )-(CH2CH2O) 1-3 -CH2CH2N((CH2CH2O) 1-3 -C 1-6 alkylene-CO2H)2, and -N(R 1c )-CH(CH2O-(CH2CH2O) 0-2 -C 1-6 alkylene-CO2H)2 selected from the group consisting of, R at each occurrence 1c and R 1d is independently hydrogen or C 1-4 alkyl, R at each occurrence 1e is independently -C 1-4 alkylene-CO2H, -C 1-4 alkylene-CONH2, or -C 1-4 alkylene-OH, R at each occurrence 1f is independently hydrogen, C 1-6 alkyl, or C 1-4 alkylene-CO2H, n at each occurrence is independently 0, 1, 2, or 3, L at each occurrence 2 is independently selected from the group consisting of -C(O)- and C 1-3 alkylene, G at each occurrence 1 is independently optionally substituted heterocyclyl. 【0081】 In some embodiments of the trans-cyclooctene modified TPD conjugates disclosed herein, the trans-cyclooctene moiety is of formula XI, 【Chemical formula】 wherein, R 2 is -OH, 2-aminoethanesulfonic acid, an N-linked natural or unnatural amino acid, or an optionally substituted ethylenediamine, and R 2 is optionally further substituted with a polyether. 【0082】 In some embodiments of the trans-cyclooctene modified TPD conjugates disclosed herein, the trans-cyclooctene moiety is of the following formula: [ka] 【0083】 In some embodiments of the trans-cyclooctene modified TPD conjugates disclosed herein, at least one trans-cyclooctene moiety is [ka] That is the case. 【0084】 In some embodiments of the trans-cyclooctene modified TPD conjugates disclosed herein, at least one trans-cyclooctene moiety is [ka] That is the case. 【0085】 In some embodiments of the trans-cyclooctene modified TPD conjugates disclosed herein, at least one trans-cyclooctene moiety is [ka] In the formula, A is an amino acid side chain (for example, C 1-4 These are alkyl and aryl (phenyl, etc.) groups, and each can be optionally substituted with -OH, -SH, -SCH3, -C(O)OH, -NH2, -NHNH2NH, -C(O)NH2, etc. 【0086】 In some embodiments of the trans-cyclooctene modified TPD conjugates disclosed herein, at least one trans-cyclooctene moiety is [ka] That is the case. 【0087】 In some embodiments of the trans-cyclooctene modified TPD conjugates disclosed herein, the trans-cyclooctene moiety is covalently bonded to the TPD via a linker. Preferred linkers are described below. 【0088】 In some embodiments of the trans-cyclooctene modified TPD conjugates disclosed herein, the linker is a cleavable linker. 【0089】 In some embodiments, the linker comprises one or more of the following: carbamates (-NHC(O)O-), esters, and amides, hydrazones, hydrazides, disulfides, N-succinimidyl-4-(2-pyridyldithio)pentanoate (SPP), N-succinimidyl-4-(2-pyridyldithio)butyrate (SPDB), 4-(4'-acetylphenoxy)butanoic acid (AcBut), one or more linear or branched, natural or unnatural amino acids, a valine-citrulline (Val-Cit) moiety, or a phenylalanine-lysine (Phe-Lys) moiety. 【0090】 In some embodiments, the linker includes 1 to 100 linked atoms, 1 to 50 linked atoms, or 5 to 50 linked atoms, or 10 to 50 linked atoms, or 1 to 40 linked atoms, or 1 to 30 linked atoms, or 1 to 20 linked atoms, or 1 to 10 linked atoms, or 1 to 5 linked atoms, or 5 to 30 linked atoms, or 10 to 30 linked atoms, or 5 to 40 linked atoms, or 5 to 50 linked atoms, or 10 to 50 linked atoms. 【0091】 In some embodiments, the linker comprises one or more chain heteroatoms and one or more alkylene, alkenylene, alkynylene, arylene, heteroarylene, cycloalkylene, or heterocycloalkylene moieties, each alkylene, alkenylene, alkynylene, arylene, heteroarylene, cycloalkylene, or heterocycloalkylene moiety independently and optionally oxo, halo, C 1-4 Alkyl, C 1-4 Alkoxy, and C 1-4 It may be substituted with 1 to 5 substituents independently selected from the haloalkyl group. 【0092】 In some embodiments, the linker is an incuttable linker. 【0093】 In some embodiments, the linker is a severable linker. 【0094】 In some embodiments, the linker contains one or more amino acids. 【0095】 In some embodiments, the linker includes a polypeptide. 【0096】 In some embodiments, the linker is an alkylene linker comprising, optionally, one or more -O-, -S-, amine, ester, amide, carbamate, carbonate, thio-succinimide, or ketone functional groups. 【0097】 In some embodiments, the linker is of the following formula: -Y 10 -(CHR 130 ) n’ -Y 20 -(CHR 140 ) n’’ -Y 30 -(CHR 150 ) m’’ -Y 40 - During the ceremony, Y 10 , Y 20 , Y 30, and Y 40 Each of them is independent, combined, -NR 110 -, -O-, -S(O) 0-2 -, -NR 110 C(O)-, -C(O)NR 110 -, -NR 110 S(O)2-, -S(O)2NR 110 -, -CR 120 =N-NR 110 -, -NR 110 -N=CR 120 -, -C(O)-, -OC(O)-, -OC(O)O-, -(CH2CH2O) 1-5 -, -C(O)O-, alkylene, alkenylene, alkynylene, arylene, or heteroarylene, and each alkylene, alkenylene, alkynylene, arylene, or heteroarylene can be independently and optionally oxo, halo, or C 1-4 Alkyl, C 1-4 Alkoxy, and C 1-4 It is substituted with 1 to 5 substituents independently selected from the haloalkyl group. Each R 110 Hydrogen and C are independent of each other. 1-4 Alkyl, C 1-4 They are haloalkyl, aryl, heteroaryl, cycloalkyl, or heterocyclyl. Each R 120 Hydrogen and C are independent of each other. 1-4 Alkyl, C 1-4 They are haloalkyl, aryl, heteroaryl, cycloalkyl, or heterocyclyl. Each R 130 Hydrogen and C are independent of each other. 1-4 Alkyl, C 1-4 Haloalkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, or amino acid side chains, Each R 140 Hydrogen and C are independent of each other. 1-4 Alkyl, C 1-4 Haloalkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, or amino acid side chains, Each R 150 Hydrogen and C are independent of each other. 1-4 Alkyl, C1-4 Haloalkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, or amino acid side chains, n', n'', and m'' are each independently 0, 1, 2, 3, 4, 5, 6, 7, or 8. 【0098】 In some embodiments, the linker is of the following formula: -Y 10 -(CH2) n’ -Y 20 -(CH2) m’’ -Y 30 - During the ceremony, Y 10 , Y 20 , and Y 30 Each of them is independent, combined, -NR 110 -, -O-, -S(O) 0-2 -, -NR 110 C(O)-, -C(O)NR 110 -, -NR 110 S(O)2-, -S(O)2NR 110 -, -CR 120 =N-NR 110 -, -NR 110 -N=CR 120 -, -C(O)-, -OC(O)-, -OC(O)O-, alkylene, alkenylene, alkynylene, arylene, heteroarylene, cycloalkylene, or heterocycloalkylene, and each alkylene, alkenylene, alkynylene, arylene, heteroarylene, cycloalkylene, or heterocycloalkylene can be independently and optionally oxo, halo, or C 1-4 Alkyl, C 1-4 Alkoxy, and C 1-4 It is substituted with 1 to 5 substituents independently selected from the haloalkyl group. Each R 110 Hydrogen and C are independent of each other. 1-4 Alkyl, C 1-4 They are haloalkyl, aryl, heteroaryl, cycloalkyl, or heterocyclyl. Each R 120 Hydrogen and C are independent of each other. 1-4 Alkyl, C1-4 is haloalkyl, aryl, heteroaryl, cycloalkyl, or heterocyclyl, n’ and m’’ are each independently 0, 1, 2, 3, 4, 5, 6, 7, or 8. 【0099】 In certain embodiments, each R 110 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, aryl, heteroaryl, cycloalkyl, or heterocyclyl, and each R 120 is independently hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, aryl, heteroaryl, cycloalkyl, or heterocyclyl. In certain embodiments, the linker is not a bond. 【0100】 The linker may include one or more of polyethylene glycol (e.g., PEG having an average molecular weight of 300 g / mol to 10,000 g / mol), ethylene-1,2-diylbis(methylcarbamate), arylene (e.g., phenylene), ethylene-oxy, amine, ester, amide, carbamate, ketone (i.e., formyl), or carbonate. 【0101】 In some embodiments, the linker is 【Chemical formula】 including one or more of. 【0102】 In some embodiments, the linker is 【Chemical formula】 including one or more of. 【0103】 In some embodiments, the linker is 【Chemical formula】 including one or more of. 【0104】 In some embodiments, the linker is one or more [ka] This includes. In some embodiments, the linker is one or more [ka] Includes. 【0105】 In some embodiments, the linker is one or more [ka] It is either or includes it. 【0106】 In some embodiments, the linker is one or more [ka] It is either or includes it. 【0107】 In some embodiments, the linker may be one or more natural or non-natural amino acids, which may be referred to as peptide linkers. The linker may be a peptide linker composed of a carboxylacyl unit and one or more amino acids that constitute a protein or peptide sequence. The linker may also contain self-immolating spacers that space the drug and protein peptide sequences. 【0108】 In some embodiments, the linker is "AYZX 2 A peptide-containing linker represented as "-W", or containing one such linker, where "A" is a carboxylate acyl unit, and "Y" and "Z" are each one or more natural or non-natural amino acids, together forming a peptide sequence, "X 2"and "W" is an optional additional linker having 1 to 50 linking atoms, or 5 to 10 linking atoms, or 1 to 10 linking atoms. In certain embodiments, one or more of the amino acids in the peptide linker are N-methylated. 【0109】 In some embodiments, Y may be at least one amino acid selected from the group consisting of alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, and proline. In some embodiments, Y may be at least one amino acid selected from the group consisting of phenylalanine, alanine, and valine. 【0110】 In some embodiments, Z may be at least one amino acid selected from the group consisting of alanine, lysine, lysine protected with acetyl or formyl, arginine, arginine protected with tosyl or nitro group, histidine, ornithine, ornithine protected with acetyl or formyl, and citrulline. In some embodiments, Z may be at least one amino acid selected from the group consisting of alanine, lysine, and citrulline. 【0111】 Exemplary Y-Z combinations include valine-citrulline, valine-alanine, and alanine-alanine. 【0112】 In some embodiments, A is -OC(O)-. 【0113】 In some embodiments, X 2 is -OC(O)-. 【0114】 In some embodiments, W is -OC(O)-. In some embodiments, X 2 is absent and W is -OC(O)-. 【0115】 In some embodiments, the moiety -X 2 -W is [ka] Includes. In some embodiments, part-X 2 teeth, [ka] That is the case. 【0116】 In some embodiments, -XW is [ka] That is the case. 【0117】 In some embodiments, -XW is [ka] That is the case. 【0118】 In some embodiments, the peptide linker is specifically modified so that it is selectively cleaved (e.g., enzymatically) by one or more tumor-associated proteases, thereby releasing the drug. 【0119】 In some embodiments, the peptide linker has a chain length of 2 to 4 amino acid residues (i.e., dipeptide, tripeptide, or tetrapeptide). However, it will be understood that peptide linkers with up to 5, 6, 7, or 8 amino acid residues can also be suitably used. 【0120】 In some embodiments, the peptide linker is Phe-Lys, Val-Lys, Val-Ala, Ala-Ala, Phe-Phe-Lys, D-Phe-Phe-Lys, Gly-Phe-Lys, Ala-Lys, Val-Cit, Phe-Cit, Leu-Cit, Ile-Cit, Trp-Cit, Phe-Ala, Gly-Phe-Leu-Gly [SEQ ID NO: 1], Ala-Leu-Ala-Leu [SEQ ID NO: 2], Phe-N 9 -Tosyl-Arg, or Phe-N9 It is -nitro-Arg. In certain embodiments, the peptide linker is Phe-Lys, Val-Lys, Val-Ala, Ala-Ala, Val-Val, Val-Cit, or D-Phe-L-Phe-Lys. In certain embodiments, the peptide linker is Val-Cit, Val-Ala, or Ala-Ala. 【0121】 In some embodiments, L, or linker, [ka] [ka] [ka] [ka] It is one or more of the following, or includes one of them. 【0122】 In some embodiments, the linker is [ka] Includes one or more of the following. 【0123】 In some embodiments, the linker is [ka] Includes one or more of the following. 【0124】 The aforementioned linker is an amino acid side chain present on X, such as lysine or cysteine ​​(for example, [ka] ) can be bound together. 【0125】 In some embodiments, the linker is -C(O)L 4 -or -C(O)C1-6 Alkilen C(O)L 4 -and, L 4 This is a bond, -N(R 12 )-C 2-3 Alkylene-N(R) 13 )C(O)-,-CH(NHC(O)R 14 )C 1-4 Alkylene-SSC 1-4 Alkylene-OC(O)-,-NHNHC(O)CH(NHC(O)R 15 )CH2C(O)-, -C 1-6 Alkylene-CH(G x )OC(O)-, [ka] And, R 12 , R 13 , R 14 , R 15 , and R 19 Each of them independently consists of hydrogen or C 1-4 It is alkyl, R 16 is hydrogen, C 1-4 Alkyl, -C 1-4 Alkylene-OH,-C 1-4 Alkylene-OC 1-4 Alkyl, -C 1-4 Alkylene-CO2H, or -C 1-4 It is alkylene-CONH2, G x The following are optional choices: halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 This is a phenyl compound substituted with 1 to 5 substituents independently selected from the group consisting of alkoxy, cyano, and nitro. 【0126】 In some embodiments, the linker includes a carbonyl moiety for conjugating at least one trans-cyclooctene moiety to the TPD. For example, the linker may include a polypeptide moiety (PPM) having a lysine residue and a lysine side chain, the PPM may also have additional lysine or other amino acid side chains conjugated to the carbonyl moiety. In some embodiments, the linker is [ka] It may include. 【0127】 In some embodiments, the linker is [ka] [ka] It is one or more of the following, or includes one of them. 【0128】 In some embodiments, the linker is [ka] That is the case. 【0129】 In some embodiments, the linker is one or more [ka] It is either or includes it. 【0130】 In some embodiments, the linker is [ka] It is either or includes it. 【0131】 In some embodiments, trans-cyclooctene modified TPD conjugates selected from Table 1 are provided. TIFF2026519211000039.tif235170TIFF2026519211000040.tif212170TIFF20265192110 00041.tif220170TIFF2026519211000042.tif228170TIFF2026519211000043.tif194170 【0132】 C. Tetrazine activator In certain embodiments, a system comprising a trans-cyclooctene modified TPD conjugate and a tetrazine activator as described herein is provided. The tetrazine activator may be formulated for systemic administration as a free tetrazine compound or, optionally via a linker, as a tetrazine-containing group covalently bonded to an antibody or antibody fragment portion, or may be administered via a therapeutic support composition comprising a biocompatible support and a tetrazine-containing group. 【0133】 In certain embodiments, the trans-cyclooctene modified TPD conjugate can target specific proteins on the treatment site or cells in a subject. In certain embodiments, the trans-cyclooctene moiety substantially blocks or inhibits the effective binding of a portion of the TPD to its target. Once the trans-cyclooctene modified TPD conjugate binds to the treatment site, contact with a tetrazine activator can release the trans-cyclooctene moiety, thus allowing the TPD to be exposed and bind to its target. 【0134】 In certain embodiments, the tetrazine activator is formulated for systemic administration. 【0135】 In a particular embodiment, the tetrazine activator is of formula IV, [ka] During the ceremony, Each R 20These are independently hydrogen, halo, cyano, nitro, alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S-R', C(=S)OR', C(=S)SR', C(=O)NR'R'', C(=S)NR'R'', NR'R'', Selected from the group consisting of NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR"R'', and NR'C(=S)NR"R'', each alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, aryl, heteroaryl, heterocyclyl, or cycloalkyl may optionally have 1 to 3 Z 1 It has been replaced with, each Z 1 These are independently halo, oxo, cyano, nitro, hydroxy, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S -R', C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'', NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR"R'', and NR'C(=S)NR"R'' are selected. In each occurrence, R' and R'' are independently selected from hydrogen, aryl, and alkyl. In each occurrence, R''' is independently selected from aryl and alkyl groups. 【0136】 In a particular embodiment, each R 20 These are independently phenyl, pyrimidinyl, triazinyl, oxazolyl, isoxazole, imidazolyl, oxadiazolyl, 6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidinyl, 5,6,7,8-tetrahydropyrido[4,3-d]pyrimidinyl, or 5,6,7,8-tetrahydropyrido[3,4-d]pyrimidinyl, each independently and optionally comprising 1 to 3 Z 1 It has been replaced with. 【0137】 In a particular embodiment, the tetrazine activator is [ka] That is the case. 【0138】 In a particular embodiment, the tetrazine activator is [ka] That is the case. 【0139】 In certain embodiments, the tetrazine activator includes a biocompatible support, an antibody or antibody fragment, or, in certain embodiments, an antibody or antibody fragment covalently bound to one or more tetrazine portions. 【0140】 In certain embodiments, the tetrazine activator is of formula I, formula II, or formula V, [ka] During the ceremony, Ring A is an aryl, cycloalkyl, heterocyclyl, or heteroaryl ring. The dotted line is R 3 and R4 An additional bond that forms tetrazine when both are absent, or R 3 and R 4 R represents an additional bond that forms dihydrotetrazine when both are present, however, when ring A is aryl. 3 and R 4 Assuming that both exist, X is a biocompatible support, antibody, or antibody fragment portion. p is between 1 and 150. In each appearance, L is independently a linker. R in each appearance 1 These are independently hydrogen, halo, cyano, nitro, alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S-R', C(=S)OR', C(=S)SR', C(=O)NR'R'', C(=S)NR'R'', NR'R'', Selected from the group consisting of NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR"R'', and NR'C(=S)NR"R'', each alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, aryl, heteroaryl, heterocyclyl, or cycloalkyl is optionally associated with 1 to 3 Z 1 It has been replaced with, R in each appearance 2These are independently halo, cyano, nitro, hydroxy, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, -C(=O)-alkyl, -C(=O)-haloalkyl, -C(=O)-alkenyl, -C(=O)-alkynyl, -C(=O)-alkoxy, -C(=O)-haloalkoxy, -C(=O)-heteroalkyl, -C(=O)-aryl, -C(=O)-heteroaryl, -C(=O)-heterocyclyl, or -C(=O)-cycloalkyl, and each alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, heteroalkyl, aryl, heteroaryl, heterocyclyl, or cycloalkyl is optionally one to three Z 1 It has been replaced with, R 3 and R 4 Both are absent, or R 3 and R 4 Each of these is a group that can be removed independently after hydrogen or a trigger event. R in each appearance 20 These are independently hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R”', SC(=O)R'”, OC(=S)R”', SC(=S)R”', S(=O)R', S(=O)2R”', S(=O)2NR'R”, C(=O)O-R', C(=O)S-R', Selected from the group consisting of C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'', NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR"R'', and NR'C(=S)NR"R'', R in each appearance 22It is independently and optionally comprises a linker of 1 to 100 linked atoms containing one or more ethylene-oxy, amine, ester, amide, carbamate, carbonate, or ketone functional groups. R in each appearance 30 These are independently halogens, cyano, nitro, hydroxy, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, or cycloalkenyl. R a , R 31a , and R 31b Each of these is independently hydrogen, C1-C6-alkyl, or C1-C6-haloalkyl. each Z 1 These are independently halo, oxo, cyano, nitro, hydroxy, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S -R', C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'', NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR"R'', and NR'C(=S)NR"R'' are selected. In each occurrence, R' and R'' are independently selected from hydrogen, aryl, and alkyl. In each occurrence, R''' is independently selected from aryl and alkyl groups. In each occurrence, t is independently 0, 1, 2, 3, or 4. 【0141】 In a particular embodiment, the tetrazine activator is of formula I, [ka] During the ceremony, X is an antibody or antibody fragment portion. p is between 1 and 16. In each appearance, L is independently a linker. R in each appearance 20 These are independently hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R”', SC(=O)R'”, OC(=S)R”', SC(=S)R”', S(=O)R', S(=O)2R”', S(=O)2NR'R”, C(=O)O-R', C(=O)S-R', Selected from the group consisting of C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'', NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR"R'', and NR'C(=S)NR"R'', R in each appearance 22 It is independently and optionally comprises a linker of 1 to 100 linked atoms containing one or more ethylene-oxy, amine, ester, amide, carbamate, carbonate, or ketone functional groups. In each occurrence, R' and R'' are independently selected from hydrogen, aryl, and alkyl. In each occurrence, R''' is independently selected from aryl and alkyl groups. 【0142】 In one embodiment, the tetrazine activator is of formula II, [ka] During the ceremony, X is an antibody or antibody fragment portion. p is between 1 and 16. In each appearance, L is independently a linker. R in each appearance 20 These are independently hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R”', SC(=O)R'”, OC(=S)R”', SC(=S)R”', S(=O)R', S(=O)2R”', S(=O)2NR'R”, C(=O)O-R', C(=O)S-R', Selected from the group consisting of C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'', NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR"R'', and NR'C(=S)NR"R'', R in each appearance 30 These are independently halogens, cyano, nitro, hydroxy, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, or cycloalkenyl. R a , R 31a , and R 31b Each of these is independently hydrogen, C1-C6-alkyl, or C1-C6-haloalkyl. In each occurrence, R' and R'' are independently selected from hydrogen, aryl, and alkyl. In each occurrence, R''' is independently selected from aryl and alkyl groups. t is independently 0, 1, 2, 3, or 4. 【0143】 One embodiment: R in each occurrence 22 These are independently linkers of 1 to 100 linked atoms and may include ethylene-oxy groups, amines, esters, amides, carbamates, carbonates, and ketone functional groups. 【0144】 In one embodiment, the tetrazine activator is of formula IIA, [ka] In the formula, L, p, X, and R 20 Each of these is independent and as defined herein. 【0145】 In one embodiment, the tetrazine activator is of formula IIB, [ka] In the formula, L, p, and X are each independently as defined herein. 【0146】 In one embodiment, the tetrazine activator is of formula IIC, [ka] In the formula, L, p, and X are each independently as defined herein. 【0147】 In one embodiment, the tetrazine activator is of formula III, [ka] During the ceremony, X is an antibody or antibody fragment portion. p is between 1 and 16. In each appearance, L is independently a linker. R in each appearance 20These are independently hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R”', SC(=O)R'”, OC(=S)R”', SC(=S)R”', S(=O)R', S(=O)2R”', S(=O)2NR'R”, C(=O)O-R', C(=O)S-R', Selected from the group consisting of C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'', NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR"R'', and NR'C(=S)NR"R'', R in each appearance 30 These are independently halogens, cyano, nitro, hydroxy, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, or cycloalkenyl. In each occurrence, R' and R'' are independently selected from hydrogen, aryl, and alkyl. In each occurrence, R''' is independently selected from aryl and alkyl groups. t is independently 0, 1, 2, 3, or 4. 【0148】 In some embodiments, the tetrazine activator is of formula IID, [ka] In the formula, X and R 20 Each of these is independently defined as herein. In some embodiments, R 20X is methyl. In some embodiments, X is an antigen-binding protein. In some embodiments, X is an antigen-binding protein that targets HER2. 【0149】 In some embodiments, the tetrazine activator is of formula IIE, [ka] In the formula, p and X are each independently as defined herein. 【0150】 In some embodiments, the tetrazine activator is of formula IIF, [ka] In the formula, p and X are each independently as defined herein. 【0151】 In some embodiments, the tetrazine activator is of formula IIG, [ka] In the formula, p and X are each independently as defined herein. 【0152】 In some embodiments of formula IIA, [ka] At least one of them is [ka] [ka] That is the case. 【0153】 In some embodiments of formula IIA, [ka] And in the formula, R20 This is defined herein. 【0154】 In some embodiments of formula IIA, [ka] That is the case. 【0155】 In some embodiments of formula IIA, [ka] And in the formula, R 20 This is defined herein. 【0156】 In some embodiments of formula IIA, [ka] That is the case. 【0157】 In some embodiments, p is 1 to 12. In some embodiments, X is an antibody. In some embodiments, p is 1 to 6, or 5 to 6.2. In some embodiments, p is 1 to 16, or 1 to 8, or 1 to 7, or 1 to 6, or 1 to 5, or 1 to 4, or 1 to 3, or 1 to 2. In some embodiments, X is an antibody fragment (e.g., Fab). 【0158】 In one embodiment, the tetrazine activator is of formula V, [ka] During the ceremony, Ring A is an aryl, cycloalkyl, heterocyclyl, or heteroaryl ring. The dotted line is R 3 and R 4 An additional bond that forms tetrazine when both are absent, or R 3 and R 4R represents an additional bond that forms dihydrotetrazine when both are present, however, when ring A is aryl. 3 and R 4 Assuming that both exist, X is a biocompatible support, antibody, or antibody fragment portion. p is between 1 and 150. In each appearance, L is independently a linker. R in each appearance 1 These are independently hydrogen, halo, cyano, nitro, alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S-R', C(=S)OR', C(=S)SR', C(=O)NR'R'', C(=S)NR'R'', NR'R'', Selected from the group consisting of NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR"R'', and NR'C(=S)NR"R'', each alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, aryl, heteroaryl, heterocyclyl, or cycloalkyl may optionally have 1 to 3 Z 1 It has been replaced with, R in each appearance 2These are independently halo, cyano, nitro, hydroxy, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, -C(=O)-alkyl, -C(=O)-haloalkyl, -C(=O)-alkenyl, -C(=O)-alkynyl, -C(=O)-alkoxy, -C(=O)-haloalkoxy, -C(=O)-heteroalkyl, -C(=O)-aryl, -C(=O)-heteroaryl, -C(=O)-heterocyclyl, or -C(=O)-cycloalkyl, and each alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, heteroalkyl, aryl, heteroaryl, heterocyclyl, or cycloalkyl is optionally one to three Z 1 It has been replaced with, R 3 and R 4 Both are absent, or R 3 and R 4 Each of these is a group that can be removed independently after hydrogen or a trigger event. each Z 1 These are independently halo, oxo, cyano, nitro, hydroxy, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S -R', C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'', NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR"R'', and NR'C(=S)NR"R'' are selected. In each occurrence, R' and R'' are independently selected from hydrogen, aryl, and alkyl. In each occurrence, R''' is independently selected from aryl and alkyl groups. In each occurrence, t is independently 0, 1, 2, 3, or 4. 【0159】 In some embodiments, the tetrazine activator is of formula V, [ka] During the ceremony, Ring A is a cycloalkyl, heterocyclyl, or heteroaryl ring. The dotted line is R 3 and R 4 If both are absent, then tetrazine or R 3 and R 4 This represents an additional bond that forms dihydrotetrazine when both are present. X is a biocompatible support, antibody, or antibody fragment portion. p is between 1 and 150. In each appearance, L is independently a linker. R in each appearance 1These are independently hydrogen, halo, cyano, nitro, alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S-R', C(=S)OR', C(=S)SR', C(=O)NR'R'', C(=S)NR'R'', NR'R'', Selected from the group consisting of NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR"R'', and NR'C(=S)NR"R'', each alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, aryl, heteroaryl, heterocyclyl, or cycloalkyl may optionally have 1 to 3 Z 1 It has been replaced with, R in each appearance 2 These are independently halo, cyano, nitro, hydroxy, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, -C(=O)-alkyl, -C(=O)-haloalkyl, -C(=O)-alkenyl, -C(=O)-alkynyl, -C(=O)-alkoxy, -C(=O)-haloalkoxy, -C(=O)-heteroalkyl, -C(=O)-aryl, -C(=O)-heteroaryl, -C(=O)-heterocyclyl, or -C(=O)-cycloalkyl, and each alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, heteroalkyl, aryl, heteroaryl, heterocyclyl, or cycloalkyl is optionally one to three Z 1 It has been replaced with, R 3 and R 4 Both are either absent or R 3It is a base that can be removed after the trigger event, R 4 is hydrogen or R 3 And, each Z 1 These are independently halo, oxo, cyano, nitro, hydroxy, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, heteroalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S -R', C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'', NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR"R'', and NR'C(=S)NR"R'' are selected. In each occurrence, R' and R'' are independently selected from hydrogen, aryl, and alkyl. In each occurrence, R''' is independently selected from aryl and alkyl groups. In each occurrence, t is independently 0, 1, 2, 3, or 4. 【0160】 In some embodiments, the tetrazine activator is of formula VI, [ka] In the formula, R 1 , R 2 , R 3 , R 4 Each of the rings A, L, p, t, and X is independently defined as herein. 【0161】 In some embodiments, R 4 It is hydrogen. 【0162】 In some embodiments, R 3 This is a base that can be removed after the trigger event. In some embodiments, the trigger event occurs in vivo. When the trigger event occurs, R 3 When removed, the dihydrotetrazine-containing group is oxidized to obtain tetrazine as shown in formula VII. [ka] In the formula, R 1 , R 2 Each of the rings A, L, p, t, and X is independently defined as herein. 【0163】 The trigger event is initiated after the administration of the tetrazine activator to the target and can be initiated by any means such as internal means (e.g., via enzymatic cleavage of functional groups, optionally followed by degradation) or external means (e.g., a photocleavable linker). In some embodiments, R 3 This includes a targeted portion such as an antibody or antibody fragment as described herein. 【0164】 In some embodiments, R 3 It contains an amino acid sequence specific to cleavage by proteases or esterases. 【0165】 In some embodiments, R 3 As shown in Table 1A, it contains an amino acid sequence specific to cleavage by proteases. TIFF2026519211000069.tif212170TIFF2026519211000070.tif229170 【0166】 In some embodiments, R 3 This includes an amino acid sequence specific to cleavage by cathepsin, matrix metalloproteinase (MMP), or PSMA. For example, in some embodiments, R 3This includes Val-Ala, Val-Cit, Ala-Ala, Phe-Lys, Lys-Lys, Phe-Arg, or Gly-Gly-Gly for cathepsin-mediated cleavage. In some embodiments, R 3 This comprises Ac-γE-PLG-S(OBn)YL or Ac-PLG-HofOrnL, where Hof is homophenylalanine and Orn is ornithine for cleavage by MMP. In some embodiments, R 3 It contains the amino acid sequence shown in Table 1B. TIFF2026519211000071.tif226170TIFF2026519211000072.tif233170TIFF2026519211000073.tif233170TIFF2026519211000074.tif231170TIFF2026519211000075.tif61170↓ indicates the cutting site Abbreviations for special amino acids: Cit: Citrulline, Cha: β-Cyclohexylalanine, Hof: Homophenylalanine, Nva: Aminosuberic acid, Dpa: D-Phenylalanine, Nle: Norleucine, Smc: S-Methylcysteine *The list of multiple amino acids before, between, or after the slash indicates alternative amino acids that can be substituted at that position, and "-" indicates that any amino acid can be substituted with the corresponding amino acid shown in the middle column. **x is any L-amino acid other than proline. Hy is any hydrophobic L-amino acid. γ indicates that the bond is a gamma-carboxyl bond. 【0167】 Additional cleavable groups are described in Choi, et al., Theranostics. 2012;2(2):156-178, and Table 2 therein is incorporated herein by reference. 【0168】 In some embodiments, R 3It is photodegradable. In some embodiments, the photodegradable group is unstable or decomposes upon exposure to light of a wavelength that matches the absorbance profile of the photodegradable group. 【0169】 In some embodiments, R 3 teeth, [ka] And, L 5 This is a direct link or linker, X 1 This is an optionally substituted peptide unit containing -NO2, an optionally substituted sugar moiety, or one or more natural or unnatural amino acids. 【0170】 In some embodiments, at least one of the following parts: [ka] teeth, [ka] [ka] [ka] It is expressed by an expression selected from, where R 1 , R 2 , R 3 , and R 4 Each of them is independent as defined herein, and optionally, ring A portion may be one or more R 2 It can be replaced in parts. 【0171】 In some embodiments, at least one of the following parts: [ka] teeth, [ka] It is expressed by an expression selected from, where X 2 is optionally an alkyl (e.g., methyl) substituted with PEG, amino acids, esters, amides, amines, -C(O)OH, -SO2, -SO3, -PO3, -PO4, or other solubility-enhancing substituents, and L, ring A, R 1 , R 2 Each of t, p, and X is independently defined as herein. 【0172】 In some embodiments, ring A is a cycloalkyl group. In some embodiments, ring A is a heterocyclyl group. In some embodiments, ring A is a heteroaryl group. In some embodiments, ring A is an aryl group. 【0173】 In some embodiments, ring A is pyrimidinyl, triazinyl, oxazolyl, isoxazole, imidazolyl, oxadiazolyl, 6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidinyl, 5,6,7,8-tetrahydropyrido[4,3-d]pyrimidinyl, or 5,6,7,8-tetrahydropyrido[3,4-d]pyrimidinyl. 【0174】 In some embodiments, ring A is phenyl. 【0175】 Several embodiments, R in each occurrence 1 Each is independently hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, aryl, heteroaryl, heterocyclyl, or cycloalkyl, and each alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, aryl, heteroaryl, heterocyclyl, or cycloalkyl is optionally one to three Z 1 It has been replaced with. 【0176】 Several embodiments, R in each occurrence 1 Independently, hydrogen or of any choice, 1 to 3 Z 1 It is an alkyl group substituted with [a specific compound]. 【0177】 In some embodiments, Z in each occurrence 1 These are independently selected from halo, hydroxy, alkoxy, and OC(=O)OR'. 【0178】 Several embodiments, R in each occurrence 2 R is independently halo, cyano, nitro, hydroxy, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, heteroalkyl, aryl, heteroaryl, heterocyclyl, or cycloalkyl. In some embodiments, R in each appearance 2 R is independently a halo, alkyl, or haloalkyl. In some embodiments, R in each occurrence 2 These are independently halo or alkyl. 【0179】 In some embodiments, t is 0 in each occurrence. 【0180】 In some embodiments, the tetrazine activator is of the following formula: [ka] In the formula, p and X are each independently as defined herein. In some embodiments, the tetrazine activator is of the following formula: [ka] In the formula, p and X are each independently as defined herein. 【0181】 In some embodiments, X is a biocompatible support. 【0182】 In some embodiments, ring A is something other than pyridyl. In some embodiments, ring A is something other than aryl. In some embodiments, ring A is something other than phenyl. 【0183】 In some embodiments, X is a biocompatible support comprising particles, polymers, viscous or non-viscous liquid materials, gels, hydrogels, crosslinked polymer matrices, metals, ceramics, plastics, bone graft materials, or proteins. 【0184】 In some embodiments, X is a biocompatible support comprising polysaccharide hydrogels, alginates, cellulose, hyaluronic acid, chitosan, chitosine, chitin, chondroitin sulfate, heparin, preferred sugar-based biomaterials, polyphosphazenes, polyanhydrides, polyacetals, poly(orthoesters), polyphosphoesters, polycaprolactones, polyurethanes, polylactides, polycarbonates, polyamides, polyethers, blends / composites thereof / or copolymers, collagen, gelatin, elastin, elastin-like polypeptides, albumin, fibrin, poly(gamma-glutamic acid), poly(L-lysine), poly(L-glutamic acid), or poly(aspartic acid). In some embodiments, X is a biocompatible support comprising hyaluronic acid having a molecular weight of about 5–25 kD, or 26–75 kD, or 76–200 kD, or >201 kD. 【0185】 In some embodiments, X is an antibody or an antibody fragment. 【0186】 Antibody and antibody fragment portion In certain embodiments, the targeting agent or X is CD25 (NCBI gene ID 3559), CEA (NCBI gene ID 634), CEACAM5 (NCBI gene ID 1048), ASPH (NCBI gene ID 444), EGFR (NCBI gene ID 1956), EPCAM (NCBI gene ID 4072), VEGFR (NCBI gene ID 3791), PDGFR (NCBI gene ID 5159), TROP2 (NCBI gene ID 4070), Nectin 4 (NCBI gene ID 81607), P SMA (NCBI gene ID 2346), BCMA (NCBI gene ID 608), CD22 (NCBI gene ID 933), CD20 (NCBI gene ID 920), CD19 (NCBI gene ID 930), CD79b (NCBI gene ID 974), CD38 (NCBI gene ID 952), CD45 (NCBI gene ID 5788), Endoglin (NCBI gene ID 2022), FGFR2 (NCBI gene ID 14183), C4.4A (NCBI gene ID 27076), Claudin-18.2 (NCBI gene ID 51208), MMP9 (NCBI gene ID 4318), folate receptor (NCBI gene ID 2348), DLL3 (NCBI gene ID 10683), CD138 (NCBI gene ID 6382), CD56 (NCBI gene ID 4684), CD37 (NCBI gene ID 951), CD74 (NCBI gene ID 972), mesothelin (NCBI gene ID 10232), IL-6R Gene ID 3570), SLAMF7 (NCBI gene ID 57823), BAFF (NCBI gene ID 10673), MUC1 (NCBI gene ID 4582), GPC3 (NCBI gene ID 2719), HER2 (NCBI gene ID 2064), HER3 (NCBI gene ID 2065), CD30 (NCBI gene ID 943), CD33 (NCBI gene ID 945), CD123 (NCBI gene ID 3563), GPNMB (NCBI gene ID 10457), cMET (NCBI gene ID 4233), CD142 (NCBI gene ID 2152), NaPi2B (NCBI gene ID 10568), GCC (NCBI gene ID 2984), STEAP1 (NCBI gene ID 26872), MUC16 (NCBI gene ID 94025), CD70 (NCBI gene ID 970), CD44 (NCBI gene ID 960), vWF ( This refers to an antibody or antibody fragment that targets one or more of the following: NCBI gene ID 7450, TNF (NCBI gene ID 7124), IL-6R (NCBI gene ID 3570), BCMA (NCBI gene ID 608), ADAMTS5 (NCBI gene ID 11096), CX3CR1 (NCBI gene ID 1524), CXCR4 (NCBI gene ID 7852), or TfR1 (NCBI gene ID 7037). 【0187】 In some embodiments, the targeting agent or X is HER2 (NCBI gene ID 2064), CLDN4 (NCBI gene ID 1364), TNC (NCBI gene ID 3371), FN1 (NCBI gene ID 2335), ITGAV (NCBI gene ID 3685), TACSTD2 (NCBI gene ID 4070), CD174 (NCBI gene ID 2525), GPNMB (NCBI gene ID 10457), GPC1 (NCBI gene ID 2817), ITGB6 This refers to an antibody or antibody fragment that targets (NCBI gene ID 3694), SEZ6 (NCBI gene ID 124925), SLITRK6 (NCBI gene ID 84189), NaPi-2b (NCBI gene ID 20531), ZIP6 (NCBI gene ID 25800), ROR1 (NCBI gene ID 4919), ROR2 (NCBI gene ID 4920), ANTXR1 (NCBI gene ID 84168), or FAP (NCBI gene ID 2191). 【0188】 In some embodiments, the targeting agent or X is an antibody or antibody fragment that targets CEA, CEACAM5, ASPH, EGFR, EPCAM, VEGFR, PDGFR, TROP2, Nectin 4, PSMA, BCMA, HER2, CD25, ANTXR1, or FAP. 【0189】 In some embodiments, the targeting agent or X is an antibody or antibody fragment that targets HER2, TROP2, Nectin-4, Claudin-18.2, MMP9, Mesothelin, FN1, FAP, TNC, or ECM, EPCAM, CEA, or CEACAM5. 【0190】 In some embodiments, the targeting agent or X is an antibody or antibody fragment that targets CEA, CEACAM5, ASPH, EGFR, EPCAM, VEGFR, PDGFR, TROP2, Nectin 4, PSMA, BCMA, HER2, or CD25. 【0191】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CD25, such as daclizumab, RG6292, basiliximab, or HuMax-TAC, or an antibody fragment derived therefrom. 【0192】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CEA, for example, rabetuzumab, 15-1-32, PR1A3, or cT84.66, or an antibody fragment derived therefrom. 【0193】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CEACAM5, such as tusamitiamab or CC4, or an antibody fragment derived therefrom. 【0194】 In certain embodiments, X is an antibody or antibody fragment that targets ASPH, for example, PAN-622, or an antibody fragment derived therefrom. 【0195】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets EGFR, such as cetuximab, necitumumab, nimotuzumab, matsuzumab, AMG595, depatuxizumab, dapatuxizumab, durigotuzumab, futuximab, GC1118, imugatuzumab, panitumumab, artumumab, tomzotuximab, or laprituximab, or an antibody fragment derived therefrom. 【0196】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets EPCAM, such as oportuzumab, sitatuzumab, tucotzumab, catumakisomab, edrecolomab, or adecatumumab, or an antibody fragment derived therefrom. 【0197】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets VEGFR, such as ramucizumab, ramucirumab, or vrinacimab, or an antibody fragment derived therefrom. 【0198】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets PDGFR, for example, olaratumab or ramucirumab, or an antibody fragment derived therefrom. 【0199】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets TROP2, for example, sacituzumab or Pr1E11, or an antibody fragment derived therefrom. 【0200】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets nectin 4, for example, enfortumab, or an antibody fragment derived therefrom. 【0201】 In a particular embodiment, the targeting agent or X is an antibody or antibody fragment that targets PSMA, for example, J591 or MLN591, or an antibody fragment derived therefrom. 【0202】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets BCMA, for example, verantamab, or an antibody fragment derived therefrom. 【0203】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CD22, such as moxetumomab, inotuzumab, epratuzumab, or pinatuzumab, or an antibody fragment derived therefrom. 【0204】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CD20, such as ubrituximab, ofatumumab, rituximab, obinutuzumab, tositumomab, or ibritumomab, or an antibody fragment derived therefrom. 【0205】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CD19, such as roncustuximab, XMAB-5574, MOR208, coltuximab, denintuzumab, tapritumomab, or MDX-1342, or an antibody fragment derived therefrom. 【0206】 In a particular embodiment, the targeting agent or X is an antibody or antibody fragment that targets CD79b, for example, polatuzumab, or an antibody fragment derived therefrom. 【0207】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CD38, such as isatuximab, daratumumab, MOR202, or TAK-079, or an antibody fragment derived therefrom. 【0208】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CD45, for example, I-131-BC8 or Iomab-B, or an antibody fragment derived therefrom. 【0209】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets endoglin, for example, carotuximab, or an antibody fragment derived therefrom. 【0210】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets FGFR2, for example, bemarituzumab or aplutuzumab, or an antibody fragment derived therefrom. 【0211】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets C4.4A, for example, lupalzumab, or an antibody fragment derived therefrom. 【0212】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets claudin-18.2, for example, zolbetuximab or clodiximab, or an antibody fragment derived therefrom. 【0213】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets MMP9, for example, andecaliximab, or an antibody fragment derived therefrom. 【0214】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets the folate receptor, such as milbetuximab, farletuzumab, MORAb-202, MORAb-003, or SP8166, or an antibody fragment derived therefrom. 【0215】 In a particular embodiment, the targeting agent or X is an antibody or antibody fragment that targets DLL3, for example, lovalpituzumab, or an antibody fragment derived therefrom. 【0216】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CD138, for example, indatuximab, or an antibody fragment derived therefrom. 【0217】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CD56, such as lorbotuzumab, promiximab, or an antibody fragment derived therefrom. 【0218】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CD37, for example, BI 836826, otreltuzumab, or naratuximab, or an antibody fragment derived therefrom. 【0219】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CD74, for example, milatuzumab, or an antibody fragment derived therefrom. 【0220】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets mesothelin, such as anetumab, amatuximab, or MMOT-0530A, or an antibody fragment derived therefrom. 【0221】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets IL-6R, such as tocilizumab or sarilumab, or an antibody fragment derived therefrom. 【0222】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets SLAMF7, for example, elotuzumab, or an antibody fragment derived therefrom. 【0223】 In a particular embodiment, the targeting agent or X is an antibody or antibody fragment that targets BAFF, for example, belimumab, or an antibody fragment derived therefrom. 【0224】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets MUC1, such as KL-6, MY.1E12, hMUC1-1H7, TAB004, huC242, cribatuzumab, 8HuDS6, gatipotuzumab, AR20.5, or cantuzumab, or an antibody fragment derived therefrom. 【0225】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets GPC3, such as codolituzumab, ECT204, or MDX-1414, or an antibody fragment derived therefrom. 【0226】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets HER2, such as pertuzumab, trastuzumab, or margetuximab, or an antibody fragment derived therefrom. 【0227】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets HER3, such as patritumab, cerivanthumab, lumuretuzumab, elgemtumab, AV-203, CDX-3379, or GSK284933, or an antibody fragment derived therefrom. 【0228】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CD30, for example, brentuximab, or an antibody fragment derived therefrom. 【0229】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CD33, such as gemtuzumab, BI 835858, vadasutuximab, or lintuzumab, or an antibody fragment derived therefrom. 【0230】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CD123, such as KHK2823, tacrotuzumab, or G4723A, or an antibody fragment derived therefrom. 【0231】 In a particular embodiment, the targeting agent or X is an antibody or antibody fragment that targets GPNMB, for example, glembatumumab, or an antibody fragment derived therefrom. 【0232】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets cMET, such as terisotuzumab, onarutuzumab, or SAIT301, or an antibody fragment derived therefrom. 【0233】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CD142, for example, tisotumab, or an antibody fragment derived therefrom. 【0234】 In a particular embodiment, the targeting agent or X is an antibody or antibody fragment that targets NaPi2B, for example, rifastuzumab, or an antibody fragment derived therefrom. 【0235】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets GCC, for example, indusatumab, or an antibody fragment derived therefrom. 【0236】 In a particular embodiment, the targeting agent or X is an antibody or antibody fragment that targets STEAP1, for example, bundletuzumab, or an antibody fragment derived therefrom. 【0237】 In a particular embodiment, the targeting agent or X is an antibody or antibody fragment that targets MUC16, for example, sofituzumab, or an antibody fragment derived therefrom. 【0238】 In a particular embodiment, the targeting agent or X is an antibody or antibody fragment that targets CD70, for example, borsetuzumab, or an antibody fragment derived therefrom. 【0239】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CD44, for example, vivacuzumab, or an antibody fragment derived therefrom. 【0240】 In a particular embodiment, the targeting agent or X is an antibody or antibody fragment that targets vWF, for example, caplacizumab, or an antibody fragment derived therefrom. 【0241】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets TNF, for example, ozoralizumab, V565, or PF-05230905, or an antibody fragment derived therefrom. 【0242】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets IL-6R, for example, bovalilizumab, or an antibody fragment derived therefrom. 【0243】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets BCMA, for example, LCAR-B38M, or an antibody fragment derived therefrom. 【0244】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets ADAMTS5, for example, M6495, or an antibody fragment derived therefrom. 【0245】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CX3CR1, for example, BI 655088, or an antibody fragment derived therefrom. 【0246】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets CXCR4, for example, AD-214 or ALX-0651, or an antibody fragment derived therefrom. 【0247】 In a particular embodiment, the targeting agent or X is an antibody or antibody fragment that targets TfR1, for example, TXB4, or an antibody fragment derived therefrom. 【0248】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets VEGFR, for example, CDP791, or an antibody fragment derived therefrom. 【0249】 In a particular embodiment, the targeting agent or X is an antibody or antibody fragment that targets PSMA, for example, GY1, or an antibody fragment derived therefrom. 【0250】 In a particular embodiment, the targeting agent or X is an antibody or antibody fragment that targets FN1, for example, L19 or NJB2, or an antibody fragment derived therefrom. 【0251】 In certain embodiments, the targeting agent or X is an antibody or antibody fragment that targets FAP, such as F19, OMTX005, or cibrotuzumab, or an antibody fragment derived therefrom. 【0252】 In a particular embodiment, the targeting agent or X is an antibody or antibody fragment that targets the TNC, for example, F16 or R6N, or an antibody fragment derived therefrom. 【0253】 In some embodiments, the targeting agent or X is an antibody. 【0254】 In some embodiments, the antibodies are daclizumab, RG6292, basiliximab, HuMax-TAC, rabetuzumab, 15-1-32, PR1A3, cT84.66, tusamithiamab, CC4, PAN-622, cetuximab, necitumumab, nimotuzumab, matsuzumab, AMG595, depatuxizumab, dapatuxizumab, durigotuzumab, futuximab, GC111 8. Imugatuzumab, Panitumumab, Artumumab, Tomzotuximab, Laprituximab, Oportuzumab, Sitatuzumab, Tucotsuzumab, Katsumakisomab, Edrecolomab, Adecatumumab, Ramucizumab, Ramucirumab, Valinacimab (vulinacimab), Oralatumab, Ramucirumab, Sacituzumab, Pr1E11, Enfortumab, J591, MLN591 , verantamab, moxetumomab, inotuzumab, epratuzumab, pinatuzumab, ubrituximab, ofatumumab, rituximab, obinutuzumab, tositumomab, ibritumomab, ronkastuximab, XMAB-5574, MOR208, coltuximab, denintuzumab, tapritumomab, MDX-1342, polatuzumab, isatuximab, daratumumab, MOR 202, TAK-079, I-131-BC8, Iomab-B, Carotuximab, Bemarituzumab, Apulzumab, Rupalzumab, Zolbetuximab, Claudiximab, Andecaliximab, Milbetuximab, Farletuzumab, MORAb-202, MORAb-003, SP8166, Robalpituzumab, Indatuximab, Rorbotuzumab, Promiximab, BI 836826, Otreltuzumab, Naratuximab, Miratuzumab, Anetuzumab, Amatsuximab, MMOT-0530A, Sarilumab, Elotuzumab, Belimumab, KL-6, MY.1E12, hMUC1-1H7, TAB004, huC242, Cribatuzumab, 8HuDS6, Gatipotuzumab, AR20.5. Cantuzumab, Codlituzumab, ECT204, MDX-1414, Pertuzumab, Trastuzumab, Margetuximab, Patrizumab, Cerivantuzumab, Lumuletuzumab, Elgemzumab, AV-203, CDX-3379, GSK284933, Brentuximab, Gemtuzumab, BI These include 835858, vadasutuximab, lintuzumab, KHK2823, tacrotuzumab, G4723A, grembatumumab, terisotuzumab, onarutuzumab, SAIT301, tisotuzumab, rifastuzumab, indusatuzumab, bundletuzumab, sofituzumab, borsetuzumab, vibatuzumab, caplacizumab, ozoralizumab, V565, PF-05230905, bovalilizumab, LCAR-B38M, BI 655088, AD-214, ALX-0651, TXB4, CDP791, GY1, L19, NJB2, F19, OMTX005, sibrotuzumab, F16, or R6N. 【0255】 In some embodiments, the targeting agent or X is an antibody selected from atezolizumab, avelumab, bevacizumab, semiprimab, cetuximab, daratumumab, dinutuximab, durvalumab, elotuzumab, ipilimumab, isatuximab, mogamulizumab, necitumumab, nivolumab, obinutuzumab, ofatumumab, olaratumumab, panitumumab, pembrolizumab, pertuzumab, ramucirumab, rituximab, and trastuzumab. 【0256】 In some embodiments, the targeting agent or X is an antibody fragment portion. 【0257】 In some embodiments, the targeting agent or X, or the antibody fragment portion, is selected from the group consisting of single-stranded variable fragments (scFv), divalent (or bivalent) single-stranded variable fragments (di-scFvs, bi-scFvs), antigen-binding fragments (Fab), single-domain antibodies (sdAb), single-domain antibodies (sdAb), antigen-binding proteins, DotBody, afibody, DARPin, DART, TandAb, diabody, ribobody, centinrin, nottin, affinrin, afimer, alphabody, antikalin, atrimer, avimer, finomer, Knitz domain, obody, pronectin, repebody, and bicyclic peptides, or Humabody. 【0258】 In some embodiments, the targeting agent or X is an antibody fragment portion selected from the group consisting of single-stranded variable fragments (scFv), divalent (or bivalent) single-stranded variable fragments (di-scFvs, bi-scFvs), antigen-binding fragments (Fab), single-domain antibodies (sdAb), and divalent (or bivalent) single-domain antibodies (sdAb). 【0259】 In some embodiments, the antibody fragment portion is an antigen-binding protein, DotBody, afibody, DARPin, DART, TandAb, diabody, ribobody, centinrin, nottin, affinrin, afimer, alphabody, antikalin, atrimer, avimer, finomer, Knitz domain, obody, pronectin, repebody, bicyclic peptide, or Humabody. 【0260】 In some embodiments, the targeting agent or X, or the antibody fragment portion, is an antigen-binding fragment (Fab). The Fab is a region on the antibody that binds to the antigen and consists of one constant domain and one variable domain in each of the heavy and light chains. In some embodiments, the Fab comprises four domains: VH, CH1, VL, and CL1. In some embodiments, the Fab contains 400–500 amino acids, or 440–480 amino acids. In some embodiments, the Fab has a molecular weight of approximately 50 kDa, or 40–55 kDa, or 45–50 kDa, or 45–55 kDa. 【0261】 In some embodiments, the Fab is trastuzumab, enfortumab, brentuximab, sacituzumab, L19 (gene ID 2335) that binds to FN-1, F16 (gene ID 3371) that binds to TNC, or NJB2 (ECM-targeting sequence). 【0262】 In some embodiments, the antibody fragment portion includes one or more PEG units that can improve circulating lifetime. 【0263】 In some embodiments, the antibody fragment portion is an antigen-binding protein. The antigen-binding protein is a protein designed to be an antibody mimetic that exhibits high affinity and specificity to a given target. In some embodiments, the antigen-binding protein is a novel recombinant polypeptide consisting of an antibody variable light chain amino acid sequence (VL) linked to a variable heavy chain sequence (VH) by a peptide designed to link the carboxyl terminus of the VL sequence to the amino terminus of the VH sequence. 【0264】 In some embodiments, the antigen-binding protein is about 5–10 kDa or about 7 kDa. In some embodiments, the antigen-binding protein is about 50–80, or 60–70, or 66 amino acids long. In some embodiments, the antigen-binding protein contains cysteine ​​only at the N-terminus or C-terminus. In some embodiments, the antigen-binding protein contains cysteine ​​only at the N-terminus. In some embodiments, the antigen-binding protein contains cysteine ​​only at the C-terminus. 【0265】 In some embodiments, the antibody fragment portion is an antigen-binding protein targeting TNC, FN1, CLDN4, MMP9, EpCAM, ITGAV, CEA, CEACAM5, ASPH, EGFR, EPCAM, VEGFR, PDGFR, TROP2, Nectin 4, PSMA, BCMA, HER2, or CD25. In some embodiments, the antibody fragment portion is an antigen-binding protein targeting HER2. Antigen-binding proteins can be prepared and tested according to standard methods or purchased from commercial sources (e.g., Affilogic). 【0266】 In some embodiments, the antibody fragment portion is daclizumab, RG6292, basiliximab, HuMax-TAC, rabetuzumab, 15-1-32, PR1A3, cT84.66, tusamithiamab, CC4, PAN-622, cetuximab, necitumumab, nimotuzumab, matsuzumab, AMG595, depatuxizumab, dapatuxizumab, durigotuzumab, futuximab, gc 1118, imugatuzumab, panitumumab, artumumab, tomzotuximab, laprituximab, oportuzumab, sitatuzumab, tucotsuzumab, catumakisomab, edrecolomab, adecatumumab, ramucizumab, ramucirumab, valinacimab (vulinacimab), oraratuzumab, ramucirumab, sacituzumab, Pr1E11, enfortumab, J591, MLN5 91, verantamab, moxetumomab, inotuzumab, epratuzumab, pinatuzumab, ubrituximab, ofatumumab, rituximab, obinutuzumab, tositumomab, ibritumomab, ronkastuximab, XMAB-5574, MOR208, coltuximab, denintuzumab, tapritumomab, MDX-1342, polatuzumab, isatuximab, daratumumab, MO R202, TAK-079, I-131-BC8, Iomab-B, carotuximab, bemarituzumab, apulzumab, rupalzumab, zolbetuximab, clodiximab, andecaliximab, milbetuximab, farletuzumab, MORAb-202, MORAb-003, SP8166, lovalpituzumab, indatuximab, rorbotuzumab, promiximab, BI 836826, Otreltuzumab, Naratuximab, Miratuzumab, Anetuzumab, Amatsuximab, MMOT-0530A, Sarilumab, Elotuzumab, Belimumab, KL-6, MY.1E12, hMUC1-1H7, TAB004, huC242, Cribatuzumab, 8HuDS6, Gatipotuzumab, AR20.5. Cantuzumab, Codlituzumab, ECT204, MDX-1414, Pertuzumab, Trastuzumab, Margetuximab, Patrizumab, Cerivantuzumab, Lumuletuzumab, Elgemzumab, AV-203, CDX-3379, GSK284933, Brentuximab, Gemtuzumab, BI Derived from 835858, vadasutuximab, lintuzumab, KHK2823, tacrotuzumab, G4723A, grembatumumab, terisotuzumab, onarutuzumab, sait301, tisotuzumab, rifastuzumab, indusatuzumab, bundletuzumab, sofituzumab, borsetuzumab, vibatuzumab, caplacizumab, ozoralizumab, V565, PF-05230905, bovalilizumab, LCAR-B38M, BI 655088, AD-214, ALX-0651, TXB4, CDP791, GY1, L19, NJB2, F19, OMTX005, sibrotuzumab, F16, or R6N. 【0267】 In some embodiments, X is an antibody fragment portion derived from atezolizumab, avelumab, bevacizumab, semiprimab, cetuximab, daratumumab, dinutuximab, durvalumab, elotuzumab, ipilimumab, isatuximab, mogamulizumab, necitumumab, nivolumab, obinutuzumab, ofatumumab, olaratumumab, panitumumab, pembrolizumab, pertuzumab, ramucirumab, rituximab, or trastuzumab. 【0268】 In a particular embodiment, X is an antibody or antibody fragment that targets vWF, for example, caplacizumab. 【0269】 In certain embodiments, X is a TNF-targeting antibody or antibody fragment, such as ozoralizumab, V565, or PF-05230905. 【0270】 In a particular embodiment, X is an antibody or antibody fragment that targets IL-6R, for example, bovalilizumab. 【0271】 In a particular embodiment, X is an antibody or antibody fragment that targets BCMA, for example, LCAR-B38M. 【0272】 In a particular embodiment, X is an antibody or antibody fragment that targets ADAMTS5, for example, M6495. 【0273】 In a particular embodiment, X is an antibody or antibody fragment that targets CX3CR1, for example, BI 655088. 【0274】 In certain embodiments, X is an antibody or antibody fragment that targets CXCR4, for example, AD-214 or ALX-0651. 【0275】 In a particular embodiment, X is an antibody or antibody fragment that targets TfR1, for example, TXB4. 【0276】 In a particular embodiment, X is an antibody or antibody fragment that targets VEGFR, for example, CDP791. 【0277】 In a particular embodiment, X is an antibody or antibody fragment that targets PSMA, for example, GY1. 【0278】 In some embodiments, the antibody fragment portion is caplacizumab, ozoralizumab, V565, PF-05230905, bovalilizumab, LCAR-B38M, M6495, BI 655088, AD-214, ALX-0651, TXB4, CDP791, or GY1. 【0279】 In some embodiments, X further comprises an imaging contrast agent. In some embodiments, the imaging contrast agent is a protein. 【0280】 In some embodiments, X is a support composition. The support composition includes a support. The support may be a biocompatible support composition, i.e., compatible with the body of the subject. In some examples, the support is non-toxic to the subject and substantially does not react with the subject's tissue or biological compounds. For example, the support may be a hydrogel. The support can be implanted in the body of the subject and support a binder (e.g., a tetrazine-containing group) and the TPD after the binder has been conjugated. Typical supports include, but are not limited to, polymers, viscous or non-viscous liquid materials, gels, hydrogels, polysaccharide hydrogels, crosslinked polymer matrices, metals, ceramics, plastics, bone graft materials, alginates, cellulose, chitosan, hyaluronic acid, chondroitin sulfate, and heparin. Supports may also include particles such as nanoparticles and microparticles. 【0281】 The hydrogel may be a polysaccharide hydrogel, alginate, cellulose, hyaluronic acid, chitosan, chitosine, chitin, chondroitin sulfate, heparin, etc. Other suitable sugar-based biomaterials are described in Polymer Advanced Technology, 2014, 25, 448-460. Polymers that can be used as supports include, but are not limited to, polyphosphazenes, polyanhydrides, polyacetals, poly(orthoesters), polyphosphoesters, polycaprolactones, polyurethanes, polylactides, polycarbonates, polyamides, and polyethers, as well as blends / composites / copolymers thereof. Typical polyethers include poly(ethylene glycol) (PEG), polypropylene glycol) (PPG), and triblock Pluronic ([PEG] n -[PPG] m -[PEG] nExamples include, but are not limited to, PEG diacrylate (PEGDA) and PEG dimethacrylate (PEGDMA), where n and m are each independent integers from 1 to 100. The support may also include proteins and other poly(amino acids), such as collagen, gelatin, elastin and elastin-like polypeptides, albumin, fibrin, poly(gamma-glutamic acid), poly(L-lysine), poly(L-glutamic acid), poly(aspartic acid), etc. 【0282】 In some embodiments, the support is a hydrogel. In some embodiments, the support is an alginate. In some embodiments, the support is chitin. In some embodiments, the support is hyaluronic acid (e.g., substantially uncrosslinked non-hydrogel hyaluronic acid). In some embodiments, the support is chitosine. In some embodiments, the support is chitosan. 【0283】 In certain embodiments, the support is a particle. The particles of this disclosure may have a diameter of 2 cm or less, for example, 1.5 cm or less, or 1 cm or less, or 0.5 cm or less. For example, the particles may be nanoparticles or microparticles. Nanoparticles include particles having an average size on the nanometer scale (e.g., 1000 nm or less). Microparticles are particles having an average size on the micrometer scale (e.g., 1000 μm or less). "Average" means the arithmetic mean. In some embodiments, the nanoparticles have a diameter in the range of 1 nm to 1 μm, for example, 10 nm to 1 μm, or 25 nm to 1 μm, or 50 nm to 1 μm, or 75 nm to 1 μm, or 100 nm to 1 μm, or 150 nm to 1 μm, or 200 nm to 1 μm, or 250 nm to 1 μm, or 300 nm to 1 μm, or 350 nm to 1 μm, or 400 nm to 1 μm, or 450 nm to 1 μm, or 500 nm to 1 μm. In other embodiments, the microparticles have diameters ranging from 1 μm to 1 mm, for example, 10 μm to 1 mm, or 25 μm to 1 mm, or 50 μm to 1 mm, or 75 μm to 1 mm, or 100 μm to 1 mm, or 150 μm to 1 mm, or 200 μm to 1 mm, or 250 μm to 1 mm, or 300 μm to 1 mm, or 350 μm to 1 mm, or 400 μm to 1 mm, or 450 μm to 1 mm, or 500 μm to 1 mm. In further embodiments, small particles with diameters of about 10 to 100 nm may be assembled to form larger complexes such as clusters or assemblies of about 1 to 10 μm. The particles of this disclosure may be substantially spherical, such that the particles have a substantially circular cross-section. Other particle shapes may also be used, but are not limited to these, such as ellipsoidal, cubic, cylindrical, conical, needle-shaped, or other irregular shapes. 【0284】 "Particles" can take the form of any manufactured material, molecule, cryptophan, virus, phage, etc. Particles may be composed of materials such as metals, ceramics, plastics, glass, composites, polymers, and hydrogels, but are not limited to these. For example, particles may be made of inert materials such as alginates or iron oxides. In some embodiments, particles may be magnetic and can be formed from paramagnetic, superparamagnetic, or ferromagnetic materials, or other materials that respond to magnetic fields. Furthermore, particles may have any shape, such as spheres, rods, or asymmetric shapes. Particles, or groups of particles in a complex, may be functionalized with receptors that have a specific affinity for or interact with clinically relevant substrates. The receptors may be inherent to the particles themselves. For example, the particles themselves may be viruses or phages that have a specific affinity for a particular substrate. Additionally or alternatively, particles may be functionalized by covalently or otherwise binding or associating with receptors that specifically bind to or otherwise recognize a particular clinically relevant substrate. The functionalized receptor can be an antibody, peptide, nucleic acid, phage, bacterium, virus, or any other molecule having a defined affinity for the target substrate. Examples of materials that may be used for “particles” and / or “carriers” include polylactic acid, polyglycolic acid, PLGA polymers, alginates and alginate derivatives, gelatin, collagen, fibrin, hyaluronic acid, laminin-rich gels, agarose, natural and synthetic polysaccharides, polyamino acids, polypeptides, polyesters, polyanhydrides, polyphosphatidines, poly(vinyl alcohol), poly(alkylene oxide), poly(allylamine) (PAM), poly(acrylate), modified styrene polymers, pluronic polyols, polyoxomers, poly(uronic acid), poly(vinylpyrrolidone), and copolymers or graft copolymers of any of the above. These examples are not limited to their concentrations, their crosslinking with different agents, their administration methods, their tuned degradation profiles, and other characteristics known to those skilled in the art. 【0285】 Particles, or groups of particles in a complex, may be functionalized with a targeting agent (e.g., ligand or antibody) that specifically (or substantially specifically) binds to a target (e.g., a target receptor or cell surface target, e.g., a clinically relevant receptor or cell surface target (e.g., an antigen)). The targeting agent may be directly bound to the particle itself. The targeting agent can be an antibody, peptide, nucleic acid, phage, bacterium, virus, or any other molecule that has specific affinity for the target receptor or cell surface target. In some examples, the receptor or cell surface target is PD-1, CTLA-4, HER2 / neu, HER1 / EGFR, VEGFR, 4-1BB, GITR, CEACAM5, or other cell receptors or cell surface targets. 【0286】 In some embodiments, the targeting agent is a monoclonal antibody. The monoclonal antibody can be the entire monoclonal antibody or a fragment thereof (e.g., an antigen-binding fragment (Fab)). In certain embodiments, the targeting agent is CD25 (NCBI gene ID 3559), CEA (NCBI gene ID 634), CEACAM5 (NCBI gene ID 1048), ASPH (NCBI gene ID 444), EGFR (NCBI gene ID 1956), EPCAM (NCBI gene ID 4072), VEGFR (NCBI gene ID 3791), PDGFR (NCBI gene ID 5159), TROP2 (NCBI gene ID 4070), Nectin 4 (NCBI gene ID 81607), PSM A (NCBI gene ID 2346), BCMA (NCBI gene ID 608), CD22 (NCBI gene ID 933), CD20 (NCBI gene ID 920), CD19 (NCBI gene ID 930), CD79b (NCBI gene ID 974), CD38 (NCBI gene ID 952), CD45 (NCBI gene ID 5788), Endoglin (NCBI gene ID 2022), FGFR2 (NCBI gene ID 14183), C4.4A (NCBI gene ID 27076), Claudin-18.2 (NCBI gene ID 51208), MMP9 (NCBI gene ID 4318), folate receptor (NCBI gene ID 2348), DLL3 (NCBI gene ID 10683), CD138 (NCBI gene ID 6382), CD56 (NCBI gene ID 4684), CD37 (NCBI gene ID 951), CD74 (NCBI gene ID 972), mesothelin (NCBI gene ID 10232), IL-6R Gene ID 3570), SLAMF7 (NCBI gene ID 57823), BAFF (NCBI gene ID 10673), MUC1 (NCBI gene ID 4582), GPC3 (NCBI gene ID 2719), HER2 (NCBI gene ID 2064), HER3 (NCBI gene ID 2065), CD30 (NCBI gene ID 943), CD33 (NCBI gene ID 945), CD123 (NCBI gene ID 3563) GPNMB (NCBI gene ID 10457), cMET (NCBI gene ID 4233), CD142 (NCBI gene ID 2152), NaPi2B (NCBI gene ID 10568), GCC (NCBI gene ID 2984), STEAP1 (NCBI gene ID 26872), MUC16 (NCBI gene ID 94025), CD70 (NCBI gene ID 970), CD44 (NCBI gene ID 960), vWF This refers to an antibody or antibody fragment that targets one or more of the following: (NCBI gene ID 7450), TNF (NCBI gene ID 7124), IL-6R (NCBI gene ID 3570), BCMA (NCBI gene ID 608), ADAMTS5 (NCBI gene ID 11096), CX3CR1 (NCBI gene ID 1524), CXCR4 (NCBI gene ID 7852), or TfR1 (NCBI gene ID 7037). 【0287】 In some embodiments, the targeting agent is a monoclonal antibody. The monoclonal antibody can be the entire monoclonal antibody or a fragment thereof (e.g., an antigen-binding fragment (Fab)). In certain embodiments, the targeting agent is HER2 (NCBI gene ID 2064), CLDN4 (NCBI gene ID 1364), TNC (NCBI gene ID 3371), FN1 (NCBI gene ID 2335), ITGAV (NCBI gene ID 3685), TACSTD2 (NCBI gene ID 4070), CD174 (NCBI gene ID 2525), GPNMB (NCBI gene ID 10457), GPC1 (NCBI gene ID 2817), ITGB6 (NCBI An antibody or antibody fragment that targets one or more of the following: gene ID 3694, SEZ6 (NCBI gene ID 124925), SLITRK6 (NCBI gene ID 84189), NaPi-2b (NCBI gene ID 20531), ZIP6 (NCBI gene ID 25800), ROR1 (NCBI gene ID 4919), ROR2 (NCBI gene ID 4920), ANTXR1 (NCBI gene ID 84168), or FAP (NCBI gene ID 2191). 【0288】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD25, such as daclizumab, RG6292, basiliximab, or HuMax-TAC, or an antibody fragment derived therefrom. 【0289】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CEA, such as rabetuzumab, 15-1-32, PR1A3, or cT84.66, or an antibody fragment derived therefrom. 【0290】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CEACAM5, such as tusamitamab or CC4, or an antibody fragment derived therefrom. 【0291】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets ASPH, such as PAN-622, or an antibody fragment derived therefrom. 【0292】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets EGFR, such as cetuximab, necitumumab, nimotuzumab, matsuzumab, AMG595, depatuxizumab, dapatuxizumab, durigotuzumab, futuximab, GC1118, imugatuzumab, panitumumab, artumumab, tomzotuximab, or laprituximab, or an antibody fragment derived therefrom. 【0293】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets EPCAM, such as oportuzumab, sitatuzumab, tucotzumab, catumakisomab, edrecolomab, or adecatumumab, or an antibody fragment derived therefrom. 【0294】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets VEGFR, such as ramucizumab, ramucirumab, or vrinacimab, or an antibody fragment derived therefrom. 【0295】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets PDGFR, such as olaratumab or ramucirumab, or an antibody fragment derived therefrom. 【0296】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets TROP2, such as sacituzumab or Pr1E11, or an antibody fragment derived therefrom. 【0297】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets nectin 4, such as enfortumab or an antibody fragment derived therefrom. 【0298】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets PSMA, such as J591 or MLN591, or an antibody fragment derived therefrom. 【0299】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets BCMA, such as verantamab or an antibody fragment derived therefrom. 【0300】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD22, such as moxetumomab, inotuzumab, epratuzumab, or pinatuzumab, or an antibody fragment derived therefrom. 【0301】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD20, such as ubrituximab, ofatumumab, rituximab, obinutuzumab, tositumomab, or ibritumomab, or an antibody fragment derived therefrom. 【0302】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD19, such as roncustuximab, XMAB-5574, MOR208, coltuximab, denintuzumab, tapritumomab, or MDX-1342, or an antibody fragment derived therefrom. 【0303】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD79b, such as polatuzumab or an antibody fragment derived therefrom. 【0304】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD38, such as isatuximab, daratumumab, MOR202, or TAK-079, or an antibody fragment derived therefrom. 【0305】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD45, such as I-131-BC8 or Iomab-B, or an antibody fragment derived therefrom. 【0306】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets endoglin, such as carotuximab or an antibody fragment derived therefrom. 【0307】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets FGFR2, such as bemarituzumab or aplutuzumab, or an antibody fragment derived therefrom. 【0308】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets C4.4A, such as lupalzumab or an antibody fragment derived therefrom. 【0309】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets claudin-18.2, such as zolbetuximab or clodiximab, or an antibody fragment derived therefrom. 【0310】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets MMP9, such as andecaliximab, or an antibody fragment derived therefrom. 【0311】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets the folate receptor, such as milbetuximab, farletuzumab, MORAb-202, MORAb-003, or SP8166, or an antibody fragment derived therefrom. 【0312】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets DLL3, such as lovalpituzumab or an antibody fragment derived therefrom. 【0313】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD138, such as indatuximab or an antibody fragment derived therefrom. 【0314】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD56, such as lorbotuzumab, promiximab, or an antibody fragment derived therefrom. 【0315】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD37, such as BI 836826, otreltuzumab, or naratuximab, or an antibody fragment derived therefrom. 【0316】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD74, such as milatuzumab or an antibody fragment derived therefrom. 【0317】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets mesothelin, such as anetumab, amatuximab, or MMOT-0530A, or an antibody fragment derived therefrom. 【0318】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets IL-6R, such as tocilizumab or sarilumab, or an antibody fragment derived therefrom. 【0319】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets SLAMF7, such as elotuzumab or an antibody fragment derived therefrom. 【0320】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets BAFF, such as belimumab or a fragment thereof. 【0321】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets MUC1, such as KL-6, MY.1E12, hMUC1-1H7, TAB004, huC242, cribatuzumab, 8HuDS6, gatipotuzumab, AR20.5, or cantuzumab, or an antibody fragment derived therefrom. 【0322】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets GPC3, such as codolituzumab, ECT204, or MDX-1414, or an antibody fragment derived therefrom. 【0323】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets HER2, such as pertuzumab, trastuzumab, or margetuximab, or an antibody fragment derived therefrom. 【0324】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets HER3, such as patritumab, cerivanthumab, lumuretuzumab, elgemtumab, AV-203, CDX-3379, or GSK284933, or an antibody fragment derived therefrom. 【0325】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD30, such as brentuximab or an antibody fragment derived therefrom. 【0326】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD33, such as gemtuzumab, BI 835858, vadasutuximab, or lintuzumab, or an antibody fragment derived therefrom. 【0327】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD123, such as KHK2823, tacrotuzumab, or G4723A, or an antibody fragment derived therefrom. 【0328】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets GPNMB, such as glembatumumab, or an antibody fragment derived therefrom. 【0329】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets cMET, such as terisotuzumab, onarutuzumab, or SAIT301, or an antibody fragment derived therefrom. 【0330】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD142, such as tisotumab or an antibody fragment derived therefrom. 【0331】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets NaPi2B, such as rifastuzumab or an antibody fragment derived therefrom. 【0332】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets GCC, such as indusatumab or an antibody fragment derived therefrom. 【0333】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets STEAP1, such as bundletuzumab or an antibody fragment derived therefrom. 【0334】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets MUC16, such as sofituzumab or an antibody fragment derived therefrom. 【0335】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD70, such as borsetuzumab or an antibody fragment derived therefrom. 【0336】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CD44, such as vivacuzumab or an antibody fragment derived therefrom. 【0337】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets vWF, such as caplacizumab or an antibody fragment derived therefrom. 【0338】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets TNF, such as ozoralizumab, V565, or PF-05230905, or an antibody fragment derived therefrom. 【0339】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets IL-6R, such as bovalilizumab or an antibody fragment derived therefrom. 【0340】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets BCMA, such as LCAR-B38M, or an antibody fragment derived therefrom. 【0341】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets ADAMTS5, such as M6495, or an antibody fragment derived therefrom. 【0342】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CX3CR1, such as BI 655088, or an antibody fragment derived therefrom. 【0343】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets CXCR4, such as AD-214 or ALX-0651, or an antibody fragment derived therefrom. 【0344】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets TfR1, such as TXB4, or an antibody fragment derived therefrom. 【0345】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets VEGFR, such as CDP791, or an antibody fragment derived therefrom. 【0346】 In certain embodiments, the targeting agent is an antibody or antibody fragment that targets PSMA, such as GY1, or an antibody fragment derived therefrom. 【0347】 Other compounds or molecules, such as fluorophores or autofluorescent or luminescent markers, which can assist in particle detection (e.g., in vivo detection), may also be attached to the particles. Ligands and / or detectable labels may be directly bound to the particles or bound to the particles via bioorthogonal functional groups, as described herein. 【0348】 In certain embodiments, the support is a bone graft material, such as a bone graft substitute. The bone graft substitute is a material that is structurally similar to bone. In some examples, the bone graft substitute is bioabsorbable, allowing it to dissolve or be absorbed into the body over time. The bone graft substitute may be osteoconductive, promoting new bone formation into blood vessels and the bone graft substitute itself. In some examples, the bone graft substitute is osteoinducible, promoting new bone formation through the active recruitment of mesenchymal stem cells from surrounding tissues. Growth factors, such as osteomorphic proteins, may be included in the bone graft substitute. Examples of bone graft substitutes include, but are not limited to, hydroxyapatite, tricalcium phosphate, demineralized bone matrix, bovine collagen, calcium sulfate, calcium phosphate, cancellous bone chips, and combinations thereof. 【0349】 In some embodiments, the tetrazine activator of this disclosure comprises a support and a tetrazine-containing group covalently bonded to the support. The tetrazine-containing group may be bonded to the support on a surface of the support, for example, on a solvent-accessible surface of the support (e.g., a surface of the support in contact with the surrounding solvent). In some cases, the tetrazine-containing group is directly bonded to the support. For example, the tetrazine-containing group may be covalently bonded to the surface of the support by covalent bonds such as amide, amine, ester, carbamate, urea, thioether, thiocarbamate, thiocarbonate, and thiourea. In some examples, the tetrazine-containing group is covalently bonded to the support by an amide bond. In other examples, the tetrazine-containing group may be linked to the support via a linker. Any suitable linker can be used to link the tetrazine-containing group to the support. Typical linkers may have 1 to 100 linking atoms and may include ethylene-oxy groups, amines, esters, amides, carbamates, carbonates, and ketone functional groups. For example, a linker may have 1 to 50 linking atoms, 5 to 50 linking atoms, or 10 to 50 linking atoms. Typical linkers, though not limited to these, include those listed below. [ka] 【0350】 In a particular embodiment, the tetrazine activator comprises a support and a tetrazine-containing group of the following formula: [ka] In the formula, R 20This includes hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocyclic, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R”', SC(=O)R'”, OC(=S)R”', SC(=S)R”', S(=O)R', S(=O)2R”', S(=O)2NR'R”, C(=O)O-R', C(=O)S-R', C(=S)O-R', C(=S)S-R', C(=O)NR'R”, C(=S)NR'R'', Selected from the group consisting of NR'R”, NR'C(=O)R”, NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR”, NR'C(=S)SR”, OC(=O)NR'R”, SC(=O)NR'R”, OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR”R”, and NR'C(=S)NR”R'', where R' and R'' in each occurrence are independently selected from hydrogen, aryl, and alkyl, and R'' in each occurrence are independently selected from aryl and alkyl, R 30 R is a halogen, cyano, nitro, hydroxy, alkyl, haloalkyl; alkenyl, alkynyl, alkoxy; haloalkoxy; heteroalkyl, aryl, heteroaryl, heterocyclic, cycloalkyl, or cycloalkenyl. a , R 31a , and R 31b Each of these is independently hydrogen, C1-C6-alkyl, or C1-C6-haloalkyl, and t is 0, 1, 2, 3, or 4. 【0351】 In a particular embodiment, the tetrazine activator has the following formula: [ka] During the ceremony, R 20This includes hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocyclic, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R”', SC(=O)R'”, OC(=S)R”', SC(=S)R”', S(=O)R', S(=O)2R”', S(=O)2NR'R”, C(=O)O-R', C(=O)S-R', C(=S)O-R', C(=S)S-R', C(=O)NR'R”, C(=S)NR'R'', Selected from the group consisting of NR'R”, NR'C(=O)R”, NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR”, NR'C(=S)SR”, OC(=O)NR'R”, SC(=O)NR'R”, OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR”R”, and NR'C(=S)NR”R'', where R' and R'' in each occurrence are independently selected from hydrogen, aryl, and alkyl, and R'' in each occurrence are independently selected from aryl and alkyl, R 22 The linker has 1 to 100 linked atoms and may contain ethylene-oxy groups, amines, esters, amides, carbamates, carbonates, and ketone functional groups. For example, the linker may have 1 to 50 linked atoms, or 5 to 50 linked atoms, or 10 to 50 linked atoms. 【0352】 In a particular embodiment, the tetrazine activator has the following formula: [ka] During the ceremony, R 20This includes hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocyclic, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R”', SC(=O)R'”, OC(=S)R”', SC(=S)R”', S(=O)R', S(=O)2R”', S(=O)2NR'R”, C(=O)O-R', C(=O)S-R', C(=S)O-R', C(=S)S-R', C(=O)NR'R”, C(=S)NR'R'', Selected from the group consisting of NR'R”, NR'C(=O)R”, NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR”, NR'C(=S)SR”, OC(=O)NR'R”, SC(=O)NR'R”, OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR”R”, and NR'C(=S)NR”R'', where R' and R'' in each occurrence are independently selected from hydrogen, aryl, and alkyl, and R'' in each occurrence are independently selected from aryl and alkyl, R 30 R is a halogen, cyano, nitro, hydroxy, alkyl, haloalkyl; alkenyl, alkynyl, alkoxy; haloalkoxy; heteroalkyl, aryl, heteroaryl, heterocyclic, cycloalkyl, or cycloalkenyl. a , R 31a , and R 31b Each of these is independently hydrogen, C1-C6-alkyl, or C1-C6-haloalkyl, and t is 0, 1, 2, 3, or 4. 【0353】 In certain embodiments, the tetrazine activator comprises a substituted alginate having units of the following formula: [ka] or its salt In the formula, R 20This includes hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocyclic, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R”', SC(=O)R'”, OC(=S)R”', SC(=S)R”', S(=O)R', S(=O)2R”', S(=O)2NR'R”, C(=O)O-R', C(=O)S-R', C(=S)O-R', C(=S)S-R', C(=O)NR'R”, C(=S)NR'R'', Selected from the group consisting of NR'R”, NR'C(=O)R”, NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR”, NR'C(=S)SR”, OC(=O)NR'R”, SC(=O)NR'R”, OC(=S)R'R'''', SC(=S)R'R'', NR'C(=O)NR”R”, and NR'C(=S)NR”R'', where R' and R'' in each occurrence are independently selected from hydrogen, aryl, and alkyl, and R'' in each occurrence are independently selected from aryl and alkyl. 【0354】 In a particular embodiment, the tetrazine activator is given by the following formula: [ka] Includes units of. 【0355】 In some embodiments, the tetrazine activator is given by the following formula: [ka] Includes units of. 【0356】 In some embodiments, the tetrazine activator is given by the following formula: [ka] Includes units of. 【0357】 In some embodiments, the tetrazine activator comprises a substituted hyaluronic acid having units of the following formula: [ka] In the formula, G 2 teeth, [ka] And R 22 It is a linker of 1 to 100 linked atoms, R 20 This is defined herein. 【0358】 In further embodiments, G 2 teeth, [ka] In further embodiments, G 2 teeth, [ka] And R 20 is hydrogen or C 1-4 It is alkyl. 【0359】 In some embodiments, the tetrazine activator is given by the following formula: [ka] The formula includes the unit R 20This includes hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocyclic, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R”', SC(=O)R'”, OC(=S)R”', SC(=S)R”', S(=O)R', S(=O)2R”', S(=O)2NR'R”, C(=O)O-R', C(=O)S-R', C(=S)O-R', C(=S)S-R', C(=O)NR'R”, C(=S)NR'R'', NR'R”, Selected from the group consisting of NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR"R'', and NR'C(=S)NR"R'', where R' and R'' in each occurrence are independently selected from hydrogen, aryl, and alkyl, and R'' in each occurrence are independently selected from aryl and alkyl. In some embodiments, R 20 is hydrogen or C 1-4 It is alkyl. 【0360】 In some embodiments, the tetrazine activator is given by the following formula: [ka] Includes units of. 【0361】 Additional tetrazine activators are exemplified in WO2017 / 044983, WO / 2015 / 139025A1, and WO / 2014 / 205126A1, the entire contents of each thereof are incorporated herein by reference. 【0362】 Hyaluronic acid derivatives include hyaluronic acid having multiple glucuronic acid units and tetrazine-containing groups linked or directly conjugated to the glucuronic acid units of hyaluronic acid. Hyaluronic acid may also have multiple N-acetylglucosamine units. In certain embodiments, the N-acetylglucosamine units of hyaluronic acid are not linked or conjugated to the tetrazine-containing groups. 【0363】 Tetrazine-containing groups can be linked or directly linked by carboxylic acids of glucuronic acid units. Tetrazine-containing groups can be incorporated into hyaluronic acid in amounts ranging from approximately 0.1% to approximately 80%, as measured by the percentage of carboxylic acids linked or conjugated to the tetrazine-containing groups, such as approximately 1% to approximately 75%, approximately 5% to approximately 75%, approximately 10% to approximately 50%, or approximately 40% to approximately 75%. 【0364】 In the tetrazine activators disclosed herein, the linker may be linear or branched (including, but not limited to, divalent, trivalent, and tetravalent branching). Thus, in a single tetrazine-based targeting agent, there may be one or more targeting moieties covalently bonded to one or more tetrazine-containing groups. In the formulas disclosed herein, "L" may be inside or outside the notation "p", however, the linker may be linear having one bond to a tetrazine-containing group and one bond to a targeting moiety, and it is intended that there may be two or more [tetrazine-L]- moieties bonded to a single targeting moiety, or the linker may be branched to have two tetrazine moieties and one bond to a targeting moiety. 【0365】 In some embodiments, L, or the linker, comprises one or more of the following: hydrazone, hydrazide, disulfide, N-succinimidyl-4-(2-pyridyldithio)pentanoate (SPP), N-succinimidyl-4-(2-pyridyldithio)butyrate (SPDB), 4-(4'-acetylphenoxy)butanoic acid (AcBut), one or more linear or branched, natural or unnatural amino acids, a valine-citrulline (Val-Cit) moiety, or a phenylalanine-lysine (Phe-Lys) moiety. 【0366】 In some embodiments, L, or the linker, comprises 1 to 100 linked atoms, 1 to 50 linked atoms, or 5 to 50 linked atoms, or 10 to 50 linked atoms, or 1 to 40 linked atoms, or 1 to 30 linked atoms, or 1 to 20 linked atoms, or 1 to 10 linked atoms, or 1 to 5 linked atoms, or 5 to 30 linked atoms, or 10 to 30 linked atoms, or 5 to 40 linked atoms, or 5 to 50 linked atoms, or 10 to 50 linked atoms. 【0367】 In some embodiments, L, or linker, comprises one or more chain heteroatoms and one or more alkylene, alkenylene, alkynylene, arylene, heteroarylene, cycloalkylene, or heterocycloalkylene moieties, each alkylene, alkenylene, alkynylene, arylene, heteroarylene, cycloalkylene, or heterocycloalkylene moiety independently and optionally oxo, halo, C 1-4 Alkyl, C 1-4 Alkoxy, and C 1-4 It may be substituted with 1 to 5 substituents independently selected from the haloalkyl group. 【0368】 In some embodiments, L, or the linker, is bound to X via a cysteine ​​or lysine residue on X. 【0369】 In some embodiments, L, or the linker, is an uncuttable linker. 【0370】 In some embodiments, L, or the linker, is a severable linker. 【0371】 In some embodiments, L, or the linker, comprises one or more amino acids. 【0372】 In some embodiments, L, or the linker, comprises a polypeptide. 【0373】 In some embodiments, L, or the linker, is an alkylene linker comprising, optionally, one or more -O-, -S-, amine, ester, amide, carbamate, carbonate, thio-succinimide, or ketone functional groups. 【0374】 In some embodiments, L, or linker, is of the following formula: -Y 10 -(CHR 130 ) n’ -Y 20 -(CHR 140 ) n’’ -Y 30 -(CHR 150 ) m’’ -Y 40 - During the ceremony, Y 10 , Y 20 , Y 30 , and Y 40 Each of them is independent, combined, -NR 110 -, -O-, -S(O) 0-2 -, -NR 110 C(O)-, -C(O)NR 110 -, -NR 110 S(O)2-, -S(O)2NR 110 -, -CR 120 =N-NR 110 -, -NR 110 -N=CR 120 -, -C(O)-, -OC(O)-, -OC(O)O-, -(CH2CH2O) 1-5-, -C(O)O-, alkylene, alkenylene, alkynylene, arylene, or heteroarylene, and each alkylene, alkenylene, alkynylene, arylene, or heteroarylene can be independently and optionally oxo, halo, or C 1-4 Alkyl, C 1-4 Alkoxy, and C 1-4 It is substituted with 1 to 5 substituents independently selected from the haloalkyl group. Each R 110 Hydrogen and C are independent of each other. 1-4 Alkyl, C 1-4 They are haloalkyl, aryl, heteroaryl, cycloalkyl, or heterocyclyl. Each R 120 Hydrogen and C are independent of each other. 1-4 Alkyl, C 1-4 They are haloalkyl, aryl, heteroaryl, cycloalkyl, or heterocyclyl. Each R 130 Hydrogen and C are independent of each other. 1-4 Alkyl, C 1-4 Haloalkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, or amino acid side chains, Each R 140 Hydrogen and C are independent of each other. 1-4 Alkyl, C 1-4 Haloalkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, or amino acid side chains, Each R 150 Hydrogen and C are independent of each other. 1-4 Alkyl, C 1-4 Haloalkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, or amino acid side chains, n', n'', and m'' are each independently 0, 1, 2, 3, 4, 5, 6, 7, or 8. 【0375】 In some embodiments, L, or linker, is of the following formula: -Y 10 -(CH2) n’ -Y 20 -(CH2) m’’ -Y30 - During the ceremony, Y 10 , Y 20 , and Y 30 Each of them is independent, combined, -NR 110 -, -O-, -S(O) 0-2 -, -NR 110 C(O)-, -C(O)NR 110 -, -NR 110 S(O)2-, -S(O)2NR 110 -, -CR 120 =N-NR 110 -, -NR 110 -N=CR 120 -, -C(O)-, -OC(O)-, -OC(O)O-, alkylene, alkenylene, alkynylene, arylene, heteroarylene, cycloalkylene, or heterocycloalkylene, and each alkylene, alkenylene, alkynylene, arylene, heteroarylene, cycloalkylene, or heterocycloalkylene can be independently and optionally oxo, halo, or C 1-4 Alkyl, C 1-4 Alkoxy, and C 1-4 It is substituted with 1 to 5 substituents independently selected from the haloalkyl group. Each R 110 Hydrogen and C are independent of each other. 1-4 Alkyl, C 1-4 They are haloalkyl, aryl, heteroaryl, cycloalkyl, or heterocyclyl. Each R 120 Hydrogen and C are independent of each other. 1-4 Alkyl, C 1-4 They are haloalkyl, aryl, heteroaryl, cycloalkyl, or heterocyclyl. n' and m'' are independently 0, 1, 2, 3, 4, 5, 6, 7, or 8. 【0376】 In a particular embodiment, each R 110 Hydrogen and C are independent of each other. 1-4 Alkyl, C 1-4 Haloalkyl, aryl, heteroaryl, cycloalkyl, or heterocyclyl, each R 120Hydrogen and C are independent of each other. 1-4 Alkyl, C 1-4 It is a haloalkyl, aryl, heteroaryl, cycloalkyl, or heterocyclyl. In certain embodiments, the linker is not a bond. 【0377】 L, or linker, may contain one or more of the following: polyethylene glycol (e.g., PEG having an average molecular weight of 300 g / mol to 10,000 g / mol), ethylene-1,2-diyrbis(methyl carbamate), arylene (ee, phenylene), ethylene-oxy, amine, ester, amide, carbamate, ketone (i.e., formyl), or carbonate. 【0378】 In some embodiments, the linker is [ka] Includes one or more of the following. 【0379】 In some embodiments, the linker is [ka] Includes one or more of the following. 【0380】 In some embodiments, the linker is [ka] Includes one or more of the following. 【0381】 In some embodiments, the linker is one or more [ka] This includes. In some embodiments, the linker is one or more [ka] Includes. 【0382】 In some embodiments, the linker is one or more [ka] It is either or includes it. 【0383】 In some embodiments, the linker is one or more [ka] It is either or includes it. 【0384】 In some embodiments, the linker may be one or more natural or non-natural amino acids, which may be referred to as peptide linkers. The linker may be a peptide linker composed of a carboxylacyl unit and one or more amino acids that constitute a protein or peptide sequence. The linker may also contain self-immolating spacers that space the drug and protein peptide sequences. 【0385】 In some embodiments, the linker is "AYZX 2 A peptide-containing linker represented as "-W", or containing one such linker, where "A" is a carboxylate acyl unit, and "Y" and "Z" are each one or more natural or non-natural amino acids, together forming a peptide sequence, "X 2 " and "W" are optional additional linkers having 1 to 50 linking atoms, or 5 to 10 linking atoms, or 1 to 10 linking atoms. In certain embodiments, one or more amino acids in the peptide linker are N-methylated. 【0386】 In some embodiments, Y may be at least one amino acid selected from the group consisting of alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, and proline. In some embodiments, Y may be at least one amino acid selected from the group consisting of phenylalanine, alanine, and valine. 【0387】 In some embodiments, Z may be at least one amino acid selected from the group consisting of alanine, lysine, acetyl or formyl-protected lysine, arginine, tosyl or nitro-protected arginine, histidine, ornithine, acetyl or formyl-protected ornithine, and citrulline. In some embodiments, Z may be at least one amino acid selected from the group consisting of alanine, lysine, and citrulline. 【0388】 Examples of YZ combinations include valine-citrulline, valine-alanine, and alanine-alanine. 【0389】 In some embodiments, A is -OC(O)-. 【0390】 In some embodiments, X 2 It is -OC(O)-. 【0391】 In some embodiments, W is -OC(O)-. In some embodiments, X 2 It is absent, and W is -OC(O)-. 【0392】 In some embodiments, part-X 2 -W is, [ka] Includes. In some embodiments, part-X 2 teeth, [ka] That is the case. 【0393】 In some embodiments, -XW is [ka] That is the case. 【0394】 In some embodiments, -XW is [ka] That is the case. 【0395】 In some embodiments, the peptide linker is specifically modified so that it is selectively cleaved (e.g., enzymatically) by one or more tumor-associated proteases, thereby releasing the drug. 【0396】 In some embodiments, the peptide linker has a chain length of 2 to 4 amino acid residues (i.e., dipeptide, tripeptide, or tetrapeptide). However, it will be understood that peptide linkers with up to 5, 6, 7, or 8 amino acid residues can also be suitably used. 【0397】 In some embodiments, the peptide linker is Phe-Lys, Val-Lys, Val-Ala, Ala-Ala, Phe-Phe-Lys, D-Phe-Phe-Lys, Gly-Phe-Lys, Ala-Lys, Val-Cit, Phe-Cit, Leu-Cit, Ile-Cit, Trp-Cit, Phe-Ala, Gly-Phe-Leu-Gly [SEQ ID NO: 1], Ala-Leu-Ala-Leu [SEQ ID NO: 2], Phe-N 9 -Tosyl-Arg, or Phe-N 9 It is -nitro-Arg. In certain embodiments, the peptide linker is Phe-Lys, Val-Lys, Val-Ala, Ala-Ala, Val-Val, Val-Cit, or D-Phe-L-Phe-Lys. In certain embodiments, the peptide linker is Val-Cit, Val-Ala, or Ala-Ala. 【0398】 In some embodiments, L, or linker, [ka] [ka] [ka] [ka] It is one or more of the following, or includes one of them. 【0399】 In some embodiments, the linker is [ka] Includes one or more of the following. 【0400】 In some embodiments, the linker is [ka] Includes one or more of the following. 【0401】 The aforementioned linker is an amino acid side chain present on X, such as lysine or cysteine ​​(for example, [ka] ) can be bound together. 【0402】 In some embodiments, the linker is -C(O)L 4 -or -C(O)C 1-6 Alkilen C(O)L 4 -and, L 4 This is a bond, -N(R 12 )-C 2-3 Alkylene-N(R) 13 )C(O)-,-CH(NHC(O)R 14 )C 1-4 Alkylene-SSC 1-4 Alkylene-OC(O)-,-NHNHC(O)CH(NHC(O)R 15 )CH2C(O)-, -C 1-6 Alkylene-CH(G x )OC(O)-, [ka] And, R 12 , R 13 , R 14 , R 15 , and R 19 Each of them independently consists of hydrogen or C 1-4 It is alkyl, R 16 is hydrogen, C 1-4 Alkyl, -C 1-4 Alkylene-OH,-C 1-4 Alkylene-OC 1-4 Alkyl, -C 1-4 Alkylene-CO2H, or -C 1-4 It is alkylene-CONH2, G x The following are optional choices: halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 This is a phenyl compound substituted with 1 to 5 substituents independently selected from the group consisting of alkoxy, cyano, and nitro. 【0403】 In some embodiments, the linker includes a carbonyl moiety for conjugating a tetrazine-containing group to the linker or X. For example, the linker may include a polypeptide moiety (PPM) having a lysine residue and a lysine side chain, and the PPM may also include additional lysine or other amino acid side chains conjugated to the carbonyl moiety. In some embodiments, the linker is [ka] It may include. 【0404】 In some embodiments, the linker is [ka] [ka] It is one or more of the following, or includes one of them. 【0405】 In some embodiments, the linker is [ka] That is the case. 【0406】 In some embodiments, the linker is [ka] It is one or more of the following, or includes one of them. 【0407】 D.TPD precursor In certain embodiments, compounds comprising TPD precursors are provided herein, which react individually in vivo at the time of administration to produce TPD. Thus, in certain embodiments, a system comprising a trans-cyclooctene-modified E3 ubiquitin ligase ligand and a tetrazine-modified target protein ligand is provided. A system comprising a tetrazine-modified E3 ubiquitin ligase ligand and a trans-cyclooctene-modified target protein ligand is also provided. 【0408】 In a particular embodiment, a method is provided for producing a TPD at a target site in a subject, comprising administering a therapeutically effective amount of a trans-cyclooctene-modified E3 ubiquitin ligase ligand and a tetrazine-modified target protein ligand to the subject. 【0409】 In a particular embodiment, a method is provided for producing a TPD at a target site in a subject, comprising administering a therapeutically effective amount of a tetrazine-modified E3 ubiquitin ligase ligand and a trans-cyclooctene-modified target protein ligand to the subject. 【0410】 In a particular embodiment, a method is provided for delivering TPD to a target site in a subject, comprising administering a therapeutically effective amount of trans-cyclooctene-modified E3 ubiquitin ligase ligand and tetrazine-modified target protein ligand to the subject. 【0411】 In a particular embodiment, a method is provided for delivering TPD to a target site in a subject, comprising administering a therapeutically effective amount of tetrazine-modified E3 ubiquitin ligase ligand and trans-cyclooctene-modified target protein ligand to the subject. 【0412】 In certain embodiments, methods are provided for treating cancer or enhancing or inducing an immune response, the methods comprising administering to a subject in need thereof a therapeutically effective amount of a trans-cyclooctene-modified E3 ubiquitin ligase ligand and a tetrazine-modified target protein ligand, or a tetrazine-modified E3 ubiquitin ligase ligand and a trans-cyclooctene-modified target protein ligand. 【0413】 In a particular embodiment, the compounds listed in Table 2 are provided. In a particular embodiment, a kit containing two or more compounds from Table 2 is provided. TIFF2026519211000123.tif202170TIFF2026519211000124.tif206170TIFF2026519211000125.tif216170 【0414】 E. Treatment method Aspects of this disclosure include methods for delivering TPD to a target location in an object. In certain embodiments, the method includes selectively delivering TPD to a target location in an object. Selective delivery of TPD can be achieved by using tetrazine activators described herein. 【0415】 In some examples, the tetrazine activators of this disclosure may be localized to a desired target site in a subject. For example, the method of this disclosure may include administering the tetrazine activator described herein to a subject. The tetrazine activator may be administered to the subject at a desired target site in the subject. In some examples, the tetrazine activator may be injected topically into the subject at a desired target site in the subject. In some embodiments, the tetrazine activator is administered systemically. In these embodiments, the tetrazine activator may be localized to a desired target site in the subject by specific binding of the tetrazine activator to its target (e.g., antibody-antigen interaction), or it may be localized on the surface of a desired target (e.g., cell surface) by specific binding of the tetrazine activator to its target (e.g., antibody-antigen interaction). 【0416】 As described herein, selective binding can occur between bioorthogonal binding partners (e.g., between tetrazine of a tetrazine activator and its complementary trans-cyclooctene of a trans-cyclooctene-modified TPD conjugate). Administration of the tetrazine activator and the resulting localization to a desired location in the target area allows the selective binding of trans-cyclooctene on the trans-cyclooctene-modified TPD conjugate between tetrazine and its complementary binder to localize the TPD to a desired target location. 【0417】 A method for treating cancer is provided herein, comprising administering to a subject in need of cancer treatment a therapeutically effective amount of a tetrazine activator described herein, or a pharmaceutically acceptable salt thereof, and a trans-cyclooctene modified TPD conjugate described herein. 【0418】 In some embodiments, the cancer is metastatic. In some embodiments, the cancer is melanoma, kidney cancer, prostate cancer, ovarian cancer, endometrial cancer, breast cancer, glioblastoma, lung cancer, soft tissue sarcoma, fibrosarcoma, osteosarcoma, pancreatic cancer, gastric cancer, squamous cell carcinoma of the head and neck, anal cancer / vulvar cancer, esophageal cancer, pancreatic adenocarcinoma, cervical cancer, hepatocellular carcinoma, Kaposi's sarcoma, non-Hodgkin lymphoma, Hodgkin lymphoma, Wilms' tumor / neuroblastoma, bladder cancer, thyroid adenocarcinoma, pancreatic neuroendocrine tumor, prostate adenocarcinoma, nasopharyngeal cancer, or cutaneous T-cell lymphoma. 【0419】 In some embodiments, the cancer is melanoma, kidney cancer, prostate cancer, ovarian cancer, breast cancer, glioma, lung cancer, soft tissue cancer, soft tissue sarcoma, osteosarcoma, or pancreatic cancer. 【0420】 In some embodiments, cancer is a solid tumor. 【0421】 In some embodiments, the cancer is a soft tissue sarcoma. 【0422】 In some embodiments, the soft tissue sarcoma is a fibrosarcoma, rhabdomyosarcoma, or Ewing's sarcoma. 【0423】 In some embodiments, the method also includes enhancing or inducing an immune response. In some embodiments, the immune response is an increase in one or more of the following: leukocytes, lymphocytes, monocytes, and eosinophils. 【0424】 In some embodiments, the method further comprises administering a therapeutically effective amount of an additional therapeutic agent selected from the group consisting of anticancer agents, immunomodulators, or trans-cyclooctenprodrugs thereof. Anticancer agents, immunomodulators, and trans-cyclooctenprodrugs thereof are known in the art. 【0425】 The indications for this approach include both hematological and solid tumors. In certain embodiments, this approach can be used for the treatment and / or diagnosis of soft tissue sarcomas: rhabdomyosarcoma, fibrosarcoma, Ewing's sarcoma, and all different subtypes of soft tissue sarcomas and osteosarcomas. The composition may be for the treatment and / or diagnosis of pigmented villonodular synovitis. 【0426】 In certain embodiments, the approach can be used to treat and / or diagnose hematological malignancies such as myelodysplastic syndrome, acute myeloid leukemia, chronic myeloid leukemia, chronic myelomonocytic leukemia, primary myelofibrosis, diffuse large B-cell lymphoma, chronic lymphocytic leukemia, monoclonal gammaglobulinemia, plasmacytomyeloma, follicular lymphoma, marginal zone lymphoma, classical Hodgkin lymphoma, monoclonal B-cell lymphocytosis, lymphoproliferative disorder NOS, T-cell lymphoma, precursor B-lymphoblastic leukemia, mantle cell lymphoma, plasmacytoma, Burkitt lymphoma, T-cell leukemia, hairy cell leukemia, precursor T-lymphoblastic leukemia, nodular lymphocyte-predominant Hodgkin lymphoma, and others. 【0427】 "Treatment" means that at least improvement of the symptoms associated with the pathological condition afflicting the subject is achieved, where improvement is used in a broad sense to mean at least a reduction of a parameter related to the treated condition, such as the severity of the symptoms. Therefore, treatment includes situations in which a pathological condition, or at least the symptoms associated with it, are completely inhibited (e.g., prevented from occurring) or stopped (e.g., terminated) so that the subject no longer suffers from that condition, or at least the symptoms that characterize that condition. Treatment may include inhibition, i.e., preventing the progression or further progression of clinical symptoms, e.g., alleviation or complete inhibition of an active disease. Treatment may also include alleviation, i.e., causing a regression of clinical symptoms. For example, in the context of cancer, the term "treating" includes any or all of the following: reduction of solid tumor growth, inhibition of cancer cell replication, reduction of overall tumor volume, extension of survival, and improvement of one or more cancer-related symptoms. 【0428】 The subject to be treated may be any subject in need of therapy, and hereby the subject to be treated is any subject that can be treated with the parent drug. Thus, a variety of subjects may be treated using the compositions disclosed herein. Generally, such subjects are “mammals,” and humans are among the subjects. Other subjects may include domestic pets (e.g., dogs and cats), livestock (e.g., cattle, pigs, goats, horses, etc.), rodents (e.g., mice, guinea pigs, and rats, e.g., animal models of diseases), and non-human primates (e.g., chimpanzees and monkeys). 【0429】 In certain embodiments, additional therapeutic agents and methods, though not limited to these, can be used for the treatment, prevention, and / or diagnosis of solid tumors including, but not limited to, melanoma (e.g., unresectable metastatic melanoma), kidney cancer (e.g., renal cell carcinoma), prostate cancer (e.g., metastatic castration-resistant prostate cancer), ovarian cancer (e.g., epithelial ovarian cancer such as metastatic epithelial ovarian cancer), endometrial cancer, breast cancer (e.g., triple-negative breast cancer), glioblastoma (e.g., glioblastoma multiforme), and lung cancer (e.g., non-small cell lung cancer), soft tissue sarcoma, fibrosarcoma, osteosarcoma, pancreatic cancer, gastric cancer, squamous cell carcinoma of the head and neck, anal / vulvar cancer, esophageal cancer, pancreatic adenocarcinoma, cervical cancer, hepatocellular carcinoma, Kaposi's sarcoma, non-Hodgkin lymphoma, Hodgkin lymphoma, Wilms' tumor / neuroblastoma, bladder cancer, thyroid adenocarcinoma, pancreatic neuroendocrine tumor, prostate adenocarcinoma, nasopharyngeal cancer, or cutaneous T-cell lymphoma. The disclosed approach is suitable for use as an adjuvant / neoadjuvant system. For example, the particles disclosed herein can be placed during a biopsy, and upon return of the test results, the physician can deliver the appropriate cocktail to the desired site in the body. This would minimize tumor size, particularly in the case of surgically resectable tumors. Then, at the end of surgery, the surgeon can administer additional tetrazine activators to target the surgical cavity and treat the patient with further doses of therapy (e.g., chemotherapy using the disclosed approach) to minimize the risk of any cancer cells that might have been missed at the resection margin. 【0430】 In certain embodiments, the tetrazine activator and trans-cyclooctene-modified TPD conjugate disclosed herein can be administered, allowing a physician to deliver the appropriate cocktail to a desired site in the body. This would minimize tumor size, particularly in the case of surgically resectable tumors. Then, at the end of surgery, the surgeon can administer additional tetrazine activator or trans-cyclooctene-modified TPD conjugate to target the surgical cavity, allowing the patient to be treated with further doses of therapy (e.g., chemotherapy using the disclosed approach) to minimize the risk of any cancer cells that might have been missed at the resection margin. 【0431】 In certain embodiments, the method disclosed herein provides the ability to position particles as disclosed herein during a biopsy. Upon return of the results, the physician can deliver the TPD to the biopsy site. 【0432】 In certain embodiments, the method of the present disclosure provides a physician with the ability to deliver TPDs to enhance the immune system with fewer side effects. This approach would be beneficial to the patient. TPDs treat solid tumors or specific locations. 【0433】 cancer The methods of this disclosure may be used to treat or prevent cancer, including metastatic cancer. Cancer is a group of related diseases that may include persistent growth signaling, evasion of growth inhibitors, resistance to cell death, activation of replication immortality, induction of angiogenesis, and activation of invasion and metastasis. The methods of this disclosure may enhance or induce an immune response against cancer in a subject. The immune response may result in an increase in one or more of the following: leukocytes, lymphocytes, monocytes, and eosinophils. 【0434】 Cancers that can be treated by the methods disclosed herein include, but are not limited to, astrocytoma, adrenocortical carcinoma, appendiceal cancer, basal cell carcinoma, cholangiocarcinoma, bladder cancer, bone cancer, brain cancer, brainstem cancer, brainstem glioma, breast cancer, cervical cancer, colon cancer, colorectal cancer, cutaneous T-cell lymphoma, diffuse endogenous pontine glioma, ductal carcinoma, endometrial cancer, ependymoma, Ewing's sarcoma, esophageal cancer, eye cancer, fibrosarcoma, gallbladder cancer, and gastric cancer. Cancer, gastrointestinal cancer, germ cell tumor, glioma, hepatocellular carcinoma, histiocytosis, Hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, Kaposi's sarcoma, kidney cancer, laryngeal cancer, leukemia, liver cancer, lung cancer, lymphoma, macroglobulinemia, melanoma, mesothelioma, oral cancer, multiple myeloma, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, osteosarcoma, ovarian cancer, pancreatic cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pituitary cancer, prostate cancer, rectal cancer, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, sarcoma, skin cancer, small cell lung cancer, small intestine cancer, soft tissue cancer, soft tissue sarcoma, solid tumor, squamous cell carcinoma, stomach cancer This includes cancer, T-cell lymphoma, testicular cancer, throat cancer, thymoma, thyroid cancer, gestational trophoblastic tumor, urethral cancer, uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Wilms' tumor, non-small cell lung cancer (NSCLC), diffuse large B-cell lymphoma (DLBCL), or oral-lingual squamous cell carcinoma (OTSCC). 【0435】 In some embodiments, the cancers that can be treated by the methods of the present disclosure are melanoma, kidney cancer, prostate cancer, ovarian cancer, breast cancer, glioma, lung cancer, soft tissue cancer, soft tissue sarcoma, osteosarcoma, or pancreatic cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a soft tissue cancer. In some embodiments, the cancer is a fibrosarcoma. In some embodiments, the cancer is a diffuse endogenous pontine glioma. In some embodiments, the cancer is a metastatic cancer. 【0436】 In some embodiments, cancers that can be treated by the methods of the present disclosure are hematological malignancies such as myelodysplastic syndrome, acute myeloid leukemia, chronic myeloid leukemia, chronic myelomonocytic leukemia, primary myelofibrosis, diffuse large B-cell lymphoma, chronic lymphocytic leukemia, monoclonal gammaglobulinemia, plasmacytomyeloma, follicular lymphoma, marginal zone lymphoma, classical Hodgkin lymphoma, monoclonal B-cell lymphocytosis, lymphoproliferative disorder NOS, T-cell lymphoma, precursor B-lymphoblastic leukemia, mantle cell lymphoma, plasmacytoma, Burkitt lymphoma, T-cell leukemia, hairy cell leukemia, precursor T-lymphoblastic leukemia, nodular lymphocyte-predominant Hodgkin lymphoma, and others. 【0437】 Therefore, the local release of ICD inducers using the compounds and methods of this disclosure can be beneficially combined with one or more immunomodulators. 【0438】 In certain embodiments, tetrazine-based targeting agents can be used for the treatment, prevention, and / or diagnosis of solid tumors including, but are not limited to, melanoma (e.g., unresectable metastatic melanoma), kidney cancer (e.g., renal cell carcinoma), prostate cancer (e.g., metastatic castration-resistant prostate cancer), ovarian cancer (e.g., epithelial ovarian cancer such as metastatic epithelial ovarian cancer), breast cancer (e.g., triple-negative breast cancer), glioblastoma (e.g., glioblastoma multiforme), and lung cancer (e.g., non-small cell lung cancer), soft tissue sarcoma, fibrosarcoma, osteosarcoma, and pancreatic cancer. 【0439】 The disclosed approach is suitable for use as an adjuvant / neoadjuvant system. For example, the tetrazine activator disclosed herein can be placed during a biopsy, and upon return of the study results, the physician can administer an appropriate cocktail to deliver the treatment to the desired site in the body (the compounds disclosed herein and any additional therapeutic agents of their choice). The biopsy results may indicate the amount and type of treatment to be delivered to the tumor site. For example, chemokines (agents that attract cancer cells and / or immune cells) and adjuvants to enhance the immune system with fewer side effects, as well as chemotherapy agents, can be delivered and combined with immunotherapeutic agents. 【0440】 The methods of this disclosure may include systemic or topical administration. In some embodiments, the tetrazine-based targeting agent is delivered systemically. In some embodiments, both the tetrazine activator and the trans-cyclooctene-modified TPD conjugate are delivered systemically. In some embodiments, the tetrazine activator is delivered topically. 【0441】 The disclosed compounds and compositions may be administered prior to surgical resection. The methods of this disclosure may minimize tumor size prior to surgical resection. This will minimize tumor size, particularly in the case of surgically resectable tumors. The disclosed conjugates, compounds, and compositions may be administered during surgical resection. The disclosed conjugates, compounds, and compositions may be administered after surgical resection. A tetrazine-based targeting agent may be placed around the surgical cavity at the end of surgical resection, and the target may then be treated with additional doses to minimize the risk of any cancer cells that may have been missed at the resection margin. 【0442】 The methods disclosed herein may include multiple systemic doses of a trans-cyclooctene modified TPD conjugate that focus on a single site. 【0443】 The methods described herein may be used to treat diffuse endogenous pontine glioma. Diffuse endogenous pontine glioma (DIPG) is a pediatric brainstem tumor that can be highly malignant and difficult to treat. There is no known curative treatment for DIPG, and survival rates have remained bleak for the past 40 years. Patients with DIPG have a median overall survival of only 11 months, and a 2-year survival rate of less than 10%. DIPG accounts for 75–80% of pediatric brainstem tumors, affecting an estimated 200–300 children each year in the United States. The rarity of this devastating disease and the lack of existing experimental model systems have hindered research, and survival rates have remained the same for the past 40 years. Diagnosis of DIPG may begin with clinical symptoms and may be confirmed by MRI. The disease may begin with several months of systemic symptoms, including behavioral changes and school difficulties, diplopia, abnormal or restricted eye movements, asymmetrical smiles, loss of balance, and weakness. Alternatively, severe neurological deterioration may occur more rapidly, with symptoms present for less than a month before diagnosis. Clinical examination may reveal polyneuropathy, cord signs such as hyperreflexia and clonus, and the triad of ataxia. Dilatation of the pontine portion of the brainstem can lead to obstructive hydrocephalus and increased intracranial pressure. 【0444】 The nuclei essential for life-sustaining functions such as respiration and heart rate are located in the pons, and without treatment, DIPG can impair respiration and heart rate. 【0445】 The methods disclosed herein may include multiple systemic doses of a trans-cyclooctene-modified TPD conjugate or tetrazine activator that focus on a single site. 【0446】 Mode of administration The treatment method may include any number of forms of administration of the disclosed conjugate, compound, or composition. The forms of administration may include tablets, pills, sugar-coated tablets, hard and soft gel capsules, granules, pellets, skin patches, skin creams, skin gels, aqueous, lipid, oily or other solutions, emulsions such as oil-in-water emulsions, liposomes, aqueous or oily suspensions, syrups, elixirs, solid emulsions, solid dispersions, or dispersible powders. In pharmaceutical compositions, the conjugates, compounds, or compositions disclosed herein may also be dispersed in microparticles, such as nanoparticle compositions. 【0447】 For parenteral administration, the conjugates, compounds, or compositions disclosed herein may be dissolved or suspended in a physiologically acceptable diluent such as water, a buffer, oil with or without a solubilizer, a surfactant, a dispersant, or an emulsifier. Suitable oils include, for example, olive oil, peanut oil, cottonseed oil, soybean oil, castor oil, and sesame oil. For parenteral administration, the conjugates, compounds, or compositions disclosed herein may be administered in the form of aqueous, lipid, oily, or other types of solutions or suspensions, or in the form of liposomes or nanosuspensions. 【0448】 The amount of composition administered to a subject can be initially determined based on the dose of the parent drug and / or guidance of the administration regimen. Generally, the composition can provide targeted delivery of the conjugated drug and / or extension of its serum half-life, and thus can provide at least one of a dose reduction or a reduction in the number of administrations in the administration regimen. Thus, the composition can result in a dose reduction and / or a reduction in the number of administrations in the administration regimen compared to the parent drug before it is conjugated in the composition of this disclosure. 【0449】 Pharmaceutical formulations may be provided in unit dosage forms. In such forms, the pharmaceutical formulation may be subdivided into unit doses containing an appropriate amount of the composition of the present disclosure. The unit dosage forms may be packaged formulations, and the packaging may contain discrete amounts of formulations such as packetized tablets, capsules, and powder in pouches, vials, or ampoules. 【0450】 In some embodiments, a kit is provided comprising a tetrazine-based targeting agent described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, and instructions for use thereof. 【0451】 In some embodiments, the kit further includes a prodrug. 【0452】 The compositions of this disclosure may be present in any preferred amount and may depend on a variety of factors, including but not limited to the subject's weight and age, disease state, etc. Preferred dosage ranges for the compositions of this disclosure include 0.1 mg to 10,000 mg, or 1 mg to 1,000 mg, or 10 mg to 750 mg, or 25 mg to 500 mg, or 50 mg to 250 mg. For example, preferred doses of the compositions of this disclosure include 1 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, or 1,000 mg. 【0453】 In some embodiments, multiple doses of the composition are administered. The frequency of administration of the composition can vary depending on any of several factors, such as the severity of the symptoms or the condition of the subject. For example, in some embodiments, the composition is administered once a month, twice a month, three times a month, every other week (qow), once a week (qw), twice a week (biw), three times a week (tiw), four times a week, five times a week, six times a week, every other day (qod), daily (qd), twice a day (qid), or three times a day (tid). 【0454】 The compositions of the Disclosure may be administered at any preferred frequency, interval, and duration. For example, the compositions of the Disclosure may be administered once per hour, or twice, three times, or more times per hour, once per day, or two, three times, or more times per day, or once every two, three, four, five, six, or seven days, in order to provide a subject with a desired dose level. When the compositions of the Disclosure are administered more than once per day, typical intervals include 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, and 60 minutes, as well as 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 20 hours, and 24 hours. The compositions of this disclosure may be administered once, twice, three times, or more times, over intervals of 1 hour, 1 to 6 hours, 1 to 12 hours, 1 to 24 hours, 6 to 12 hours, 12 to 24 hours, 1 day, 1 to 7 days, 1 week, 1 to 4 weeks, 1 month, 1 to 12 months, more than 1 year, or indefinitely. 【0455】 The compositions of this disclosure may be co-administered with other activators. Co-administration includes administering the compositions of this disclosure and the activators within 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 20 hours, or 24 hours from each other. Co-administration also includes administering the compositions of this disclosure and the activators simultaneously or nearly simultaneously (for example, within about 1 minute, 5 minutes, 10 minutes, 15 minutes, 20 minutes, or 30 minutes from each other), or sequentially in any order. Furthermore, each of the compositions of this disclosure and the activators may be administered once daily, or two, three, or more times daily, to provide a desired daily dose level. 【0456】 Co-administration can be achieved through co-transplantation or co-injection. 【0457】 In some embodiments, co-administration can be achieved by co-formulation, for example, by preparing a single pharmaceutical formulation containing both the composition and the activator of the Disclosure. In other embodiments, the composition and the activator of the Disclosure can be formulated separately and co-administered to a subject. 【0458】 The compositions and activators of this disclosure may be present in formulations in any suitable weight ratio, for example, 1:100 to 100:1 (w / w), or 1:50 to 50:1, or 1:25 to 25:1, or 1:10 to 10:1, or 1:5 to 5:1 (w / w). The compositions and other activators of this disclosure may be present in any suitable weight ratio, such as 1:100 (w / w), 1:75, 1:50, 1:25, 1:10, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 10:1, 25:1, 50:1, 75:1, or 100:1 (w / w). Other dosages and dosage ratios of the compositions and activators of this disclosure are preferred in formulations and methods described herein. 【0459】 Combination therapy In one embodiment, a method is provided for treating cancer or enhancing or inducing an immune response, the method comprising administering to a subject in need a therapeutically effective amount of a tetrazine-based targeting agent of the present disclosure or a pharmaceutically acceptable salt or composition thereof; and a prodrug such as those described herein; and optionally, a therapeutically effective amount of an additional therapeutic agent selected from the group consisting of anticancer agents, immunomodulators, or trans-cyclooctene prodrugs thereof. 【0460】 This disclosure also provides pharmaceutically acceptable combinations comprising tetrazine-based targeting agents described herein, or pharmaceutically acceptable salts thereof, or compositions thereof, for use in the treatment or prevention of cancer, or for use in enhancing or inducing an immune response; prodrugs described herein; and optionally, additional therapeutic agents selected from the group consisting of anticancer agents, immunomodulators, or trans-cyclooctene prodrugs thereof. 【0461】 This disclosure also provides the use of tetradine-based targeting agents described herein, or pharmaceutically acceptable salts or compositions thereof; prodrugs such as those described herein; and optionally, a therapeutically effective amount of additional therapeutic agents selected from the group consisting of anticancer agents, immunomodulators, or trans-cyclooctene prodrugs thereof, for use in the treatment or prevention of cancer, or in enhancing or inducing an immune response, in a pharmaceutically effective amount. 【0462】 In the methods and uses described herein, the components of a pharmaceutically acceptable combination may be administered / used simultaneously, separately, sequentially, and in any order, and the components may be administered separately or as a fixed combination. For example, delaying or treating disease progression according to this disclosure may include administering a first active ingredient in the form of a free salt or pharmaceutically acceptable salt, simultaneously or sequentially in any order, in a joint therapeutic effective dose or effective dose, e.g., a daily dose corresponding to the amount described herein, and administering a second active ingredient in the form of a free salt or pharmaceutically acceptable salt. The individual active ingredients of a combination may be administered separately at different times during the course of treatment, or simultaneously in divided or single dosage forms. Accordingly, this disclosure should be understood to encompass all such regimens of simultaneous or alternating treatment, and the term “administer” should be interpreted accordingly. Accordingly, as used herein, a pharmaceutically acceptable combination defines either a fixed combination of one unit dosage form or separate dosage forms for concomitant administration, and concomitant administration may be independently, simultaneously, or at different times. As a further example, tetrazine-based targeting agents (or therapeutic tetrazine-based targeting agents) and prodrugs may be administered simultaneously (e.g., via co-injection or co-transplantation), separately, or sequentially, followed by the administration / use of additional therapeutic agents selected from the group consisting of anticancer agents, their immunomodulators, or trans-cyclooctene prodrugs. 【0463】 Methods and uses in the treatment of cancer include the administration / localization of tetrazine-based targeting agents to tumors. In the methods and uses disclosed herein, the administration of prodrugs, or pharmaceutically acceptable salts or compositions thereof; tetrazine-based targeting agents; and optionally additional therapeutic agents may inhibit tumor growth. 【0464】 Additional therapeutic agents may be administered simultaneously with or sequentially to the conjugates and compositions of the Disclosure. Sequential administration includes administration before or after the conjugates and compositions of the Disclosure. Additional therapeutic agents may be administered before the conjugates and compositions of the Disclosure. Additional therapeutic agents may be administered after the conjugates and compositions of the Disclosure. Additional therapeutic agents may be administered simultaneously with the conjugates and compositions of the Disclosure. In some embodiments, additional therapeutic agents may be administered in the same composition as the conjugates of the Disclosure. In other embodiments, there may be a time interval between the administration of the additional therapeutic agent and the administration of the conjugates or compositions of the Disclosure. In some embodiments, administration of additional therapeutic agents with the conjugates or compositions of the Disclosure may allow for lower doses and / or less frequent administration of the other therapeutic agents. When used in combination with one or more other active ingredients, the conjugates or compositions of the Disclosure and the other active ingredients may be used in lower doses than when each is used alone. Therefore, the pharmaceutical compositions of this disclosure include those containing one or more other active ingredients in addition to the conjugate of this disclosure. 【0465】 anticancer drugs Examples of anticancer drugs, though not limited to these, include abiraterone acetate, Abitrexate (methotrexate), Abraxane (paclitaxel albumin-stabilized nanoparticle formulation), ABVD, ABVE, ABVE-PC, AC, AC-T, Adcetris (brentuximab vedotin), ADE, Ado-trastuzumab emtansine, Adriamycin (doxorubicin hydrochloride), Adrucil (fluorouracil), afatinib dimaleate, Afinitor (everolimus), Aldara (imiquimod), and Aldeslo Ikin, alemtuzumab, Alimta (pemetrexed disodium), Aloxi (palonosetron hydrochloride), Ambochlorin (chlorambucil), aminolevulinic acid, anastrozole, aprepitant, Aredia (pamidronate disodium), Arimidex (anastrozole), Aromasin (exemestane), Arranon (nelarabine), arsenic trioxide, Arzerra (ofatumumab), asparaginase, erwinia chrysanthemum, Avastin (bevacizumab), axitinib, azacitidine, BEA COPP, bendamustine hydrochloride, BEP, bevacizumab, bexarotene, Bexxar (tositumomab and 131-iodine tositumomab), bicalutamide, bleomycin, bortezomib, Bosulif (bosutinib), bosutinib, brentuximab vedotin, busulfan, Busulfex (busulfan), cabazitaxel, cabozantinib-S-malate, CAF, Campath (aremtuzumab), Camptosar (irinotecan hydrochloride), capecitabine, CAPOX, carboplatin, carboplatin-taxol, carfil Zomib, Casodex (bicalutamide), CeeNU (lomustine), Cerubidine (daunorubicin hydrochloride), Cervarix (recombinant HPV bivalent vaccine), cetuximab, chlorambucil, chlorambucil-prednisone, CHOP, cisplatin, Clafen (cyclophosphamide), clofarabine, Clofarex (clofarabine), Clolar (clofarabine), CMF, Cometriq (cabozantinib-S-malate), COPP, COPP-ABV, Cosmegen (dactinomycin), crizotinib,CVP, cyclophosphamide, Cyfos (ifosfamide), cytarabine, cytarabine, cytarabine liposomal, Cytosar-U (cytarabine), Cytoxan (cyclophosphamide), dabrafenib, dacarbazine, Dacogen (decitabine), dactinomycin, dasatinib, daunorubicin hydrochloride, decitabine, degarelix, denileukin difutitox, denosumab, DepoCyt (liposomal cytarabine) Bin), DepoFoam (liposomalcitarabine), dexrazoxane hydrochloride, docetaxel, Doxil (doxorubicin hydrochloride liposome), doxorubicin hydrochloride, doxorubicin hydrochloride liposome, Dox-SL (doxorubicin hydrochloride liposome), DTIC-Dome (dacarbazine), Efudex (fluorouracil), Elitek (rasburicase), Ellence (epirubicin hydrochloride), Eloxa tin (oxaliplatin), eltrombopagolamine, Emend (aprepitant), enzalutamide, epirubicin hydrochloride, EPOCH, Erbitux (cetuximab), eribulin mesylate, Erivedge (bismodegib), erlotinib hydrochloride, Erwinaze (asparaginase erwinia chrysanthemum), Etopophos (etoposide phosphate), etoposide, etoposide phosphate, Evacet ( Doxorubicin hydrochloride (liposome), everolimus, Evista (raloxifene hydrochloride), exemestane, Fareston (toremifene), Faslodex (fulvestrant), FEC, Femara (letrozole), filgrastim, Fludara (fludarabine phosphate), fludarabine phosphate, Fluoroplex (fluorouracil), fluorouracil, Folex (methotrexate), Folex PFS (methotrexate), Folfiri, Folfiri-bevacizumab, Folfiri-cetuximab, Folfirinox, Folfox (leucovorin, fluorouracil, oxaliplatin), Folotyn (pralatrexate), FU-LV, fulvestrant, Gardasil (recombinant HPV tetravalent vaccine), Gazyva (obinutuzumab), gefitinib, gemcitabine hydrochloride, gemcitabine-cisplatin, gemcitabine-oxaliplatin,Gemtuzumab ozogamicin, Gemzar (gemcitabine hydrochloride), Gilotrif (afatinib dimaleate), Gleevec (imatinib mesylate), glucarpidase, goserelin acetate, Halaven (eribulin mesylate), Herceptin (trastuzumab), HPV bivalent vaccine, recombinant HPV tetravalent vaccine, recombinant Hycamtin (topotecan hydrochloride), Hyper-CV AD, ibritumomab tiuxetan, ibrutinib, ICE, Iclusig (ponatinib hydrochloride), Ifex (ifosfamide), ifosfamide, Ifosfamidum (ifosfamide), imatinib mesylate, Imbruvica (ibrutinib), imiquimod, Inlyta (axitinib), Intron A (recombinant interferon alpha-2b), iodine-131 tositumomab and tositumoma Mab, ipilimumab, Iressa (gefitinib), irinotecan hydrochloride, Istodax (romidepsin), ixabepyrone, Ixempra (ixabepyrone), Jakafi (ruxolitinib phosphate), Jevtana (cabazitaxel), Kadcyla (Ado-trastuzumab emtansine), Keoxifene (raloxifene hydrochloride), Kepivance (palifermin), Kypro lis (carfilzomib), lapatinib ditosylate, lenalidomide, letrozole, leucovorin calcium, Leukeran (chlorambucil), leuprolide acetate, Levulan (aminolevulinic acid), Linfolizin (chlorambucil), LipoDox (doxorubicin hydrochloride liposome), liposomalcitarabine, lomustine, Lupron (leuprolide acetate), Lupron Depot (leuprolide acetate), Lupron Depot-Ped (leuprolide acetate), Lupron Depot-3 Month (leuprolide acetate), Lupron Depot-4 Month (leuprolide acetate), Marqibo (vincristine sulfate liposome), Matulane (procarbazine hydrochloride), mechloretamine hydrochloride, Megace (megestrol acetate), megestrol acetate, Mekinist (trametinib), mercaptopurine, mesna, Mesnex (mesna), Methazolastone (temozolomide),Methotrexate, Methotrexate LPF (Methotrexate), Mexate (Methotrexate), Mexate-AQ (Methotrexate), Mitomycin C, Mitozytrex (Mitomycin C), MOPP, Mozobil (Prelixafor), Mustargen (Mechloretamine Hydrochloride), Mutamycin (Mitomycin C), Myleran (Busulfan), Mylosar (Azacitidine), Mylotarg (Gemtuzumab Ozogamicin), Nanoparticle Paclitaxel (Paclitaxel Albumin Stabilized Nano (Particle formulation), Navelbine (vinorelbine tartrate), nelarabine, Neosar (cyclophosphamide), Neupogen (filgrastim), Nexavar (sorafenib tosylate), nilotinib, Nolvadex (tamoxifen citrate), Nplate (romiplostim), obinutuzumab, ofatumumab, omasetaxin mepesuccinate, Oncaspar (peguaspar gauze), Ontak (denileukin difutitox), OEPA, OPPA, oxaliplatin, paclitaxel, paclitaxel albumin Stabilized nanoparticle formulations, Palifermin, Palonosetron hydrochloride, Pamidronate disodium, Panitumumab, Paraplat (carboplatin), Paraplatin (carboplatin), Pazopanib hydrochloride, Pegaspargase, Peginterferon alfa-2b, PEG-Intron (pegylated interferon alfa-2b), Pemetrexed disodium, Perjeta (pertuzumab), Pertuzumab, Platinol (cisplatin), Platinol-AQ (cisplatin), Plerixafor, Pomalidomide, Po malyst (pomalidomide), ponatinib hydrochloride, pralatrexate, prednisone, procarbazine hydrochloride, Proleukin (aldesleukin), Prolia (denosumab), Promacta (eltrombopagolamine), Provenge (ciproisel-T), Purinethol (mercaptopurine), radium-223 dichloride, raloxifene hydrochloride, rasburicase, R-CHOP, R-CVP, recombinant HPV bivalent vaccine, recombinant HPV tetravalent vaccine, recombinant interferon alpha-2b, regorafenib,Revlimid (lenalidomide), Rheumatrex (methotrexate), Rituxan (rituximab), rituximab, romidepsin, romiplostim, rubidomycin (daunorubicin hydrochloride), ruxolitinib phosphate, Sclerosol intrapleural aerosol (talc), ciproisel-T, sorafenib tosylate, Sprycel (dasatinib), Stanford V, Sterilized talc powder (talc), Steritalc (talc), Stivarga (regorafenib), Sunitinib malate, Sutent (sunitinib malate), Sylatron (pegylated interferon alpha-2b), Synovir (thalidomide), Synribo (omacetaxin mepesuccinate), Tafinlar (dabrafenib), talc, tamoxifen citrate, Tarabine PFS (cytarabine), Tarceva (erlotinib hydrochloride), Targretin (bexarotene), Tasigna (nilotinib), Taxol (paclitaxel), Taxotere (docetaxel), Temodar (temozolomide), temozolomide, temsirolimus, thalidomide, Thalomid (thalidomide), Toposar (etoposide), topotecan hydrochloride, toremifene, Torisel (temsirolimus), tocitumomab and 1 131 Iodine tocitumomab, Totect (dexrazoxane hydrochloride), trametinib, trastuzumab, Treanda (bendamustine hydrochloride), Trisenox (arsenic trioxide), Tykerb (lapatinib ditosylate), vandetanib, VAMP, Vectibix (panitumumab), VelP, Velban (vinblastine sulfate), Velcade (bortezomib), Velsar (vinblastine sulfate), vemurafenib, VePesid (etoposide), Viadur (leuprolide acetate), Vidaza (azacitidine), vinblastine sulfate, Vincasar PFS (vincristine sulfate), vincristine sulfate, vincristine sulfate liposomes, vinorelbine tartrate, bismodegib, Voraxaze (glucarpidase), vorinostat, Votrient (pazopanib hydrochloride), Wellcovorin (leucovorin calcium), Xalkori (crizotinib), Xeloda (capecitabine),Xelox, Xgeva (denosumab), Xofigo (radium-223 dichloride), Xtandi (enzalutamide), Yervoy (ipilimumab), Zaltrap (Ziv-aflibercept), Zelboraf (vemurafenib), Zevalin (ibritumomab tiuxetan), Examples include Zinecard (dexrazoxane hydrochloride), Ziv-aflibercept, Zoladex (goserelin acetate), zoledronic acid, Zolinza (vorinostat), Zometa (zoledronic acid), and Zytiga (abiraterone acetate). 【0466】 The anticancer drugs may be PBD dimer, calicheamicin, speromycin, tubulisin B, rhizoxin, drastatin, didemnin B, camptothecin, CBI, temsirolimus, actinomycin D, epothyron B, taxol, cryptophycin, SN38, Velcade, bruseantine, DAVLBH, DM1, filantoside, alimta, T2 toxin, MMC, vantaranib, vinorelbine, breferdin, sunitinib, daunomycin, semacanib, tarceva, irressa, irinotecan, LY-541503, geldanomycin, gemcitabine, methotrexate, gleevec, topotecan, bleomycin, doxorubicin, cisplatin, N-mustard, etoposide, or 5-FU. 【0467】 In certain embodiments, the anticancer agent is an anthracycline. In certain embodiments, the anticancer agent is a taxane. In certain embodiments, the anticancer agent is gemcitabine. In certain embodiments, the anticancer agent is doxorubicin. In certain embodiments, the anticancer agent is docetaxel. In certain embodiments, the anticancer agent is SN38. In certain embodiments, the anticancer agent is monomethyl auristatin E. 【0468】 Compound Synthesis Targeted moieties may be prepared using the methods disclosed herein and modifications thereof, as will be apparent from the consideration of the methods disclosed herein and methods well known in the art. In addition to the teachings herein, conventional and well known synthetic methods may be used. The synthesis of typical targeted moieties disclosed herein can be achieved as disclosed in the following examples. Where available, reagents and starting materials may be purchased commercially from, for example, Sigma Aldrich or other chemical suppliers. 【0469】 Furthermore, conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesirable reactions. Suitable protecting groups for various functional groups, as well as preferred conditions for protecting and deprotecting specific functional groups, are well known in the art. For example, numerous protecting groups are described in Wuts, PGM, Greene, TW, & Greene, TW (2006). Greene's protective groups in organic synthesis. Hoboken, NJ, Wiley-Interscience, and the references cited therein. 【0470】 The compounds, conjugates, and activators provided herein can be prepared by methods adapted from the literature (see, for example, WO2020 / 077140, WO2018 / 187740, WO2017 / 044983, WO2015 / 139025, WO2014 / 205126, WO2022 / 032191, WO2021 / 007160, WO2020 / 077140, WO2018 / 187740, WO2017 / 044983, WO2015 / 139025, and WO2014 / 205126, the methods described herein, in their entirety). 【0471】 For example, the trans-cyclooctene modified TPD conjugate (P1) described herein can be prepared under standard coupling conditions according to the following scheme I, where A is TPD, the linker is as defined herein, TCO is at least one trans-cyclooctene moiety, Xx is a leaving group and Yy is a nucleophilic moiety, or Xx is a nucleophilic moiety and Yy is a leaving group. Scheme I [ka] 【0472】 Example 1: Synthesis of Compound 16 [ka] Intermediate 1a (0.50 g, 2.34 mmol), intermediate 2a (1.00 g, 2.34 mmol), and HATU (1.34 g, 3.51 mmol) were combined and dissolved in anhydrous CH2Cl2 (20 mL). Anhydrous DIPEA (1.63 mL, 9.36 mmol) was added, and the mixture was stirred at room temperature under an N2 atmosphere for 18 hours. The reaction mixture was directly packed into a silica column, and compound 3a was purified by flash chromatography using a gradient of MeOH in CH2Cl2 (0-6%). 【0473】 HRMS(ESI)C 32 H 44 Calculated value for N5O6S[M+1]: 626.3007; Measured value: 626.3549 【0474】 1H NMR(500MHz,MeOD)δ 8.89(s,1H),7.47(t,J=7.2Hz,2H),7.42(d,J=8.3Hz,2H),4.67(d,J=9.2Hz,1H),4.61(d,J=8.4Hz, 1H),4.58-4.56(m,1H),4.52(d,J=6.4Hz,2H),4.39(dd,J=15.5,4.5Hz,1H),4.28(dd,J=12.2,7.3H z,1H),3.87(d,J=11.1Hz,1H),3.82(dd,J=11.0,3.8Hz,1H),2.68-2.55(m,2H),2.49(s,3H),2.36( s,1H),2.23(dd,J=13.1,7.7Hz,1H),2.10(ddd,J=12.9,8.9,4.3Hz,1H),1.46(s,9H),1.05(s,9H). 【0475】 13 C NMR(126MHz,MeOD)δ 172.99,171.28,170.37,151.77,147.71,138.85,132.15,130.15,128.82,127.6 6,79.73,70.73,69.50,59.32,57.31,42.59,37.71,35.89,27.55,25.54,14.53. [ka] 【0476】 A solution of HCl (4N) in dioxane (6 mL) was cooled in an ice bath under a nitrogen atmosphere. A solution of compound 3a in CH2Cl2 (6 mL) was added, and the reaction mixture was gradually heated to room temperature and stirred for 1 hour. The solvent was evaporated under reduced pressure. The crude product was used for further synthesis without any purification. 【0477】 HRMS(ESI)C 27 H 38 ClN5O4S [M+2] 2+ Calculated value: 281.6161; Measured value: 282.3049 【0478】 11H NMR (500 MHz, MeOD) δ 9.89 (s, 1H), 7.60 - 7.57 (m, 2H), 7.54 (d, J = 8.0 Hz, 2H), 4.67 (s, 1H), 4.60 (d, J = 8.7 Hz, 1H), 4.57 (d, J = 8.0 Hz, 1H), 4.53 (s, 1H), 4.43 (d, J = 15.7 Hz, 1H), 4.22 - 4.18 (m, 1H), 3.89 (d, J = 10.8 Hz, 1H), 3.83 (dd, J = 10.8, 3.7 Hz, 1H), 3.76 (dd, J = 11.4, 5.5 Hz, 2H), 3.69 (d, J = 7.3 Hz, 8H), 3.62 - 3.59 (m, 1H), 3.51 (q, J = 7.0 Hz, 1H), 3.25 (q, J = 7.4 Hz, 1H), 2.90 - 2.84 (m, 1H), 2.79 (dd, J = 10.1, 7.5 Hz, 1H), 2.66 (s, 1H), 2.62 (s, 3H), 2.26 (dd, J = 12.9, 7.9 Hz, 1H), 2.11 (ddd, J = 13.2, 9.2, 4.3 Hz, 1H), 1.40 (t, J = 7.1 Hz, 7H), 1.20 (t, J = 7.0 Hz, 2H), 1.09 (s, 9H). 【0479】 13 13C NMR (126 MHz, MeOD) δ 173.14, 170.20, 167.13, 154.87, 141.79, 140.88, 135.64, 128.98, 127.94, 75.96, 73.79, 72.17, 71.05, 69.67, 66.79, 65.51, 60.80, 59.45, 58.25, 56.59, 54.46, 51.29, 42.38, 42.24, 37.61, 35.19, 25.65, 21.07, 17.36, 15.92, 14.06, 11.97, 11.80. 【Chem.】 【0480】 Intermediate 4a (200 mg, 0.38 mmol), compound 5a (100.05 mg, 0.38 mmol), and HATU (217.24 mg, 0.571 mmol) were combined in anhydrous CH2Cl2 (20 mL). DIPEA (0.264 mL, 1.52 mmol) was added dropwise, and the reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was loaded onto a silica column, and intermediate 6a was purified by flash chromatography using a gradient of MeOH in CH2Cl2 (0-8%). 【0481】 HRMS(ESI)C 38 H 55 Calculated value for N6O9S[M+1]: 771.3746; measured value: 771.4397 【0482】 1 H NMR(500MHz,MeOD)δ 8.90(s,1H),7.48(d,J=8.1Hz,2H),7.44(d,J=8.1Hz,2H),4.69(t,J=6.7Hz,1H),4.66(s,1H),4.60-4.55(m,2H),4.52(s,1H) ,4.38(d,J=15.5Hz,1H),4.08(d,J=2.2Hz,2H),3.87(d,J=10.8Hz,1H),3.82(dd,J=10.9,3.8Hz,1H),3.74(dd,J=5.1,3.2Hz,3 H),3.69-3.66(m,2H),3.56(t,J=5.7Hz,2H),3.37(s,2H),3.25(dd,J=11.9,6.3Hz,3H),2.71(dt,J=14.5,5.0Hz,2H),2.50(s ,4H),2.41(s,1H),2.22(d,J=7.8Hz,1H),2.10(ddd,J=13.3,9.0,4.5Hz,1H),1.45(s,9H),1.38(t,J=6.0Hz,6H),1.05(s,9H). 【0483】 13C NMR(126MHz,MeOD)δ 172.93,171.51,170.58,151.11,148.14,138.43,132.19,130.14,129.01,127.52,78.51,7 1.13,70.57,69.37,59.30,57.53,51.37,42.23,37.48,35.15,27.71,25.09,21.27,14.18. [ka] 【0484】 Intermediate 6a (0.150 g, 0.38 mmol) was dissolved in CH2Cl2 (5 mL) and TFA (5 mL). The reaction mixture was stirred at room temperature under a N2 atmosphere for 2 hours. The solvent was removed under reduced pressure, and the product was azeotropically mixed with CH2Cl2 (5 mL) to remove any residual TFA. 【0485】 1 H NMR(500MHz,MeOD)δ 9.58(s,1H),7.53(d,J=8.1Hz,2H),7.48(d,J=8.1Hz,2H),4.83(t,J=6.1Hz,1H),4.66-4.61( m,2H),4.58-4.53(m,2H),4.44-4.39(m,1H),4.09(s,2H),3.92(d,J=11.0Hz,1H),3.82(dd,J =11.0,3.4Hz,1H),3.78-3.70(m,8H),3.25-3.19(m,2H),2.75-2.62(m,2H),2.56(s,3H),2.4 0(s,1H),2.29(dd,J=12.6,7.4Hz,1H),2.16-2.08(m,1H),1.38(t,J=6.2Hz,5H),1.06(s,9H). [ka] 【0486】 Intermediate 8a (0.060 g, 0.15 mmol) and PyBOP (0.117 g, 0.225 mmol) were combined in anhydrous DMF (6 mL). DIPEA (0.104 ml, 0.6 mmol) was added dropwise, and the reaction mixture was stirred at room temperature under an N2 atmosphere for 15 minutes. Intermediate 7a (0.10 g, 0.15 mmol) was added, and the reaction mixture was stirred for 18 hours. The reaction mixture was diluted 1:1 DMSO:H2O and purified by preparative HPLC using a gradient of 30-95% CH3CN (0.1% formic acid) in H2O (0.1% formic acid). 【0487】 HRMS(ESI)C 52 H 62 ClN 10 Calculated value for O8S2[M+1]: 1053.3877; Measured value: 1053.2644 【0488】 1 H NMR(500MHz,MeOD)δ 8.90(s,1H),8.10(d,J=7.7Hz,1H),7.47(d,J=7.9Hz,4H),7.42(d,J=8.4Hz,4H),4.71(t,J=6.7Hz,1H),4.66(t,J=6.4Hz,2H),4.59(t, J=8.2Hz,1H),4.53(d,J=15.3Hz,1H),4.47(s,1H),4.37(d,J=15.4Hz,1H),4.07(d,J=3.5Hz,2H),3.87(d,J=10.8Hz,1H),3.80(dd,J=1 0.9,3.8Hz,1H),3.75(d,J=5.4Hz,2H),3.71(d,J=5.1Hz,2H),3.66(t,J=5.2Hz,2H),3.52-3.46(m,3H),3.37(dd,J=15.2,5.3Hz,1H),2 .71(s,5H),2.48(s,3H),2.46(s,3H),2.23-2.17(m,1H),2.11-2.06(m,1H),2.05(s,1H),1.71(s,3H),1.41-1.37(m,2H),1.04(s,9H). 【0489】 13C NMR(126MHz,DMSO)δ 172.32,170.15,169.80,169.74,169.55,163.57,163.51,151.98,148.18,139.94,1 37.16,135.74,132.72,131.26,130.64,130.34,130.13,130.07,129.14,128.94,12 7.93, 80.97, 73.45, 70.72, 70.25, 69.85, 69.71, 69.32, 59.21, 57.13, 56.82, 54.26, 51.19, 42.14, 40.90, 38.34, 37.91, 36.09, 26.72, 22.17, 16.39, 14.52, 13.16, 11.78. [ka] 【0490】 Intermediate 9a (0.050 g, 0.047 mmol), compound 10a (9.4 uL, 0.047 mmol), sodium ascorbate (0.002 g, 0.0094 mmol, 20 mol%), and copper(II) sulfate (0.002 g, 0.0094 mmol, 20 mol%) were combined in THF (1.5 mL). Five to six drops of H2O were added, and the reaction mixture was stirred for 16 hours. The solvent was evaporated under reduced pressure, and the mixture was redissolved in 1:1 DMSO:H2O (1.5 mL each). Intermediate 11a was purified by preparative HPLC using a gradient of 30-95% CH3CN (0.1% formic acid) in H2O (0.1% formic acid). 【0491】 HRMS(ESI)C 60 H 80 ClN 14 O 11 Calculated value for S2[M+1]: 1271.5255; measured value: 1271.2644 【0492】 11H NMR (500 MHz, MeOD) δ 8.89 (s, 1H), 8.19 (s, 3H), 7.75 (s, 1H), 7.46 (d, J = 2.6 Hz, 4H), 7.43 (d, J = 3.7 Hz, 4H), 4.86 - 4.82 (m, 1H), 4.66 (d, J = 3.3 Hz, 1H), 4.62 (s, 1H), 4.53 (d, J = 5.5 Hz, 3H), 4.49 (s, 1H), 4.41 - 4.36 (m, 1H), 4.02 (d, J = 5.3 Hz, 2H), 3.91 (t, J = 9.1 Hz, 1H), 3.85 (d, J = 4.1 Hz, 2H), 3.80 (dd, J = 10.8, 3.5 Hz, 1H), 3.68 (t, J = 6.1 Hz, 7H), 3.66 - 3.61 (m, 6H), 3.58 (d, J = 7.1 Hz, 6H), 3.49 (dd, J = 12.9, 7.1 Hz, 3H), 3.37 (dd, J = 15.2, 5.2 Hz, 1H), 3.24 (dd, J = 14.9, 6.0 Hz, 1H), 3.14 (d, J = 5.7 Hz, 3H), 2.70 (s, 3H), 2.68 (s, 1H), 2.48 (s, 3H), 2.46 (s, 3H), 2.26 (dd, J = 12.8, 7.3 Hz, 1H), 2.12 - 2.06 (m, 1H), 2.05 (s, 1H), 1.71 (s, 3H), 1.02 (s, 9H). 【0493】 13 13C NMR (126 MHz, MeOD) δ 173.01, 171.59, 171.22, 170.84, 170.45, 164.78, 164.22, 155.64, 151.48, 150.76, 147.65, 142.67, 138.89, 136.74, 136.56, 132.15, 131.98, 131.81, 130.63, 130.56, 130.12, 129.97, 128.97, 128.39, 127.90, 127.64, 127.58, 123.75, 70.77, 70.05, 69.90, 69.83, 69.67, 69.46, 69.01, 66.50, 59.47, 57.75, 56.76, 53.84, 52.31, 49.88, 42.32, 39.28, 39.16, 39.03, 37.61, 37.42, 35.35, 27.74, 25.61, 14.46, 13.04, 11.57, 10.20. 【Chem.】 【0494】 Intermediate 11a (0.090 g, 0.071 mmol), TCO-bis-NHS (0.036 g, 0.085 mmol), and DIPEA (37 mL, 0.21 mmol) were combined in DMF (5 mL). The reaction mixture was stirred for 16 hours. The solvent was evaporated under reduced pressure, and the mixture was redissolved in CH2Cl2 (3 mL). Intermediate 12a was purified by preparative TLC using 15% MeOH in CH2Cl2 as the mobile phase. 【0495】 HRMS(ESI)C 75 H 97 ClN 15 O 17 Calculated value for S2[M+1]: 1578.6311; Measured value: 1578.3682 【0496】 1 H NMR(500MHz,CDCl3)δ 8.76(s,1H),8.08-7.98(m,1H),7.89-7.85(m,1H),7.67-7.59(m,1H),7.34(s,1H),6.09-5.76(m,1H),5.71-5.47(m, 1H),5.19-5.13(m,1H),4.94-4.80(m,1H),4.75-4.62(m,1H),4.61-4.42(m,1H),4.40-4.31(m,1H),4.07-3.98(m,1H) ),3.84-3.75(m,1H),3.67(s,1H),3.56(s,1H),3.45(s,1H),3.23-3.01(m,1H),2.82(s,1H),2.50(s,1H),2.41(s,1H) ),2.34-2.20(m,1H),2.20-2.13(m,1H),2.12-1.79(m,1H),1.66(s,1H),1.25(s,1H),1.07-1.02(m,1H),0.94(s,1H). [ka] 【0497】 Intermediate 12a (0.050 g, 0.031 mmol), aspartic acid (0.021 g, 0.15 mmol), DIPEA (108 mL, 0.63 mmol), and BSA (0.063 g, 0.31 mmol) were combined in CH2Cl2 (5 mL). The reaction mixture was stirred for 16 hours. The solvent was evaporated under reduced pressure, and the mixture was redissolved in a 1:1 solution of CH3CN (4 mL) and 5% aqueous citric acid (4 mL, pH 3.3). The mixture was stirred at room temperature for 1 hour. Compound 16 was purified by preparative HPLC using a gradient of 30-95% CH3CN (0.1% formic acid) in H2O (0.1% formic acid). 【0498】 HRMS(ESI)C 75 H 99 ClN 15 O 18 Calculated value for S2[M+1]: 1596.6417; Measured value: 1596.3116 【0499】 1 H NMR(500MHz,MeOD)δ 8.88(s,1H),8.64-8.59(m,1H),8.51-8.47(m,1H),8.16-7.95(m,2H) ,7.74(s,1H),7.51-7.36(m,7H),5.95-5.86(m,1H),5.68-5.61(m,1H ),5.08(s,1H),4.69-4.59(m,4H),4.58-4.44(m,4H),4.42-4.34(m,1 H),4.06-3.98(m,2H),3.93(d,1H),3.89-3.76(m,3H),3.73-3.61(m,6 H),3.58(s,8H),3.54-3.46(m,4H),3.40-3.35(m,2H),3.30-3.10(m, 4H),2.84(t,1H),2.73-2.65(m,3H),2.49(d,5H),2.30-2.22(m,2H),2 .16-2.05(m,2H),2.02-1.94(m,2H),1.90-1.86(m,1H),1.84-1.77(m ,1H),1.72-1.68(m,3H),1.66-1.57(m,1H),1.12(s,2H),1.02(s,8H). 【0500】 Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which this disclosure pertains. 【0501】 The embodiments described herein as illustrative can also be suitably implemented without any elements, limitations, or restrictions not specifically disclosed herein. Therefore, terms such as “comprising,” “including,” and “containing” should be read broadly and non-restrictively. Furthermore, the terms and expressions used herein are for illustrative purposes only, not limitation, and the use of such terms and expressions is not intended to exclude any equivalents of the shown and described features or any part thereof, and it should be recognized that various modifications are possible within the scope of the claims. 【0502】 All publications, patent applications, patents, and other references referred to herein are expressly incorporated by reference in whole to the same extent that each is incorporated by reference individually. In case of any conflict, this specification shall prevail, including definitions. 【0503】 While this disclosure has been described in conjunction with the embodiments described above, it should be understood that the foregoing description and examples are illustrative and not intended to limit the scope of this disclosure. Other aspects, advantages, and variations within the scope of this disclosure will be obvious to those skilled in the art in which this disclosure relates.

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