Compounds containing cleavable linkers and uses thereof

Abstract

A compound comprising a cleavable linker, the use thereof, and an intermediate compound for preparing said compound are provided, and more specifically, the compound comprising a cleavable linker of the present invention may comprise an activator having a specific function or activity (e.g., a drug, a toxin, a ligand, a probe for detection, etc.), an SO2 functional group capable of selectively releasing said activator, and a functional group that induces a chemical reaction, a physicochemical reaction and / or a biological reaction by an external stimulus, and additionally may comprise a ligand having binding specificity to a desired target receptor (e.g., an oligopeptide, a polypeptide, an antibody, etc.).

Description

Technology Field

[0001] Related applications

[0002] This application claims the benefit of U.S. provisional patent application No. 62 / 788,013 filed on January 3, 2019, the contents of which are incorporated herein by reference in their entirety. Background Technology

[0003] Antibody-drug conjugates (ADCs) are emerging as a powerful class of antitumor agents with efficacy across various cancers. ADCs typically comprise three distinct features: a cell-binding agent or targeting moiety; a linker; and a cytotoxic agent. The linker component of an ADC possesses desirable target specificity, and in other words , which has high activity in tumor cells but low activity in healthy cells, is an important feature in the development of targeted anticancer drugs. The problem to be solved

[0004] Therefore, an improved linker useful for manufacturing ADCs is required. means of solving the problem

[0005] Chemical formula ( I' A conjugate of ) or a pharmaceutically acceptable salt thereof is provided herein:

[0006]

[0007] Here:

[0008] CB is a targeting moiety;

[0009] cb and n are each independently integers having a value of 1 to about 20, preferably 1 to about 10;

[0010] Each DL independently has the chemical formula ( I" It is a device having the structure of ):

[0011]

[0012] Each Q is an activator independently connected to L' by a heteroatom, preferably O or N;

[0013] Z' is the chemical formula independently in each case ( I" The structure of ) (CB) cb A linker connecting to, a solubilizing group, a reactive group (e.g., a precursor group), a solid surface (e.g., a particle), a stabilizing group, a chelator, a biopolymer (e.g., an immunoglobulin, nucleic acid, protein, oligopeptide, polypeptide, antibody, fragment or repeat of an antigenic polypeptide), an activator, or a detectable moiety, provided that at least one occurrence of Z' is of the chemical formula ( I" The structure of ) (CB) cb Connect to;

[0014] Each L' is a spacer moiety independently attached to SO2 through a heteroatom selected from O, S, and N, preferably O or N, and the cleavage of the bond between L' and SO2 is selected to facilitate the cleavage of the bond between L' and Q to release an activator;

[0015] Each X is independently -O-, -C(R b )(R c )-, or -N(R c )-, preferably -C(R b )(R c )-and;

[0016] E is an integer having 1, 2 or 3, preferably 1;

[0017] Ar is a 6-membered aryl or 6-membered heteroaryl ring;

[0018] Y' is -N(R a )-, -O-, or -S- and;

[0019] At least one X is located in an ortho relation or a para relation with respect to Y' on Ar;

[0020] TG is an inducing group that generates N, O, or S atoms capable of initiating the emission of SO2 and (Q)q-(L')w when cleaved;

[0021] Each q is an integer having a value of 1 to about 20, preferably 1 to about 10, independently;

[0022] Each w and x is an integer that independently has a value of 0 or 1;

[0023] Each R a and R c is independently hydrogen or lower alkyl; and

[0024] Each R b is independently Z', hydrogen, or lower alkyl, and at least one R b is Z' and; or

[0025] R b and R c ..., together with the atoms to which they are attached, form a 3-5-membered ring, preferably a 3-4-membered ring;

[0026] However, when w is 0, q is 1.

[0027] In a specific embodiment, each X is located in an ortho relation or a para relation with respect to Y' on Ar.

[0028] In a specific embodiment, at least one X and Y' are located in an ortho relationship with respect to each other on Ar. In such a specific embodiment, E is 2 or 3. In a specific embodiment where E is 2, the two Xs that occurred are located in an ortho relationship with respect to Y'.

[0029] In another embodiment, at least one X and Y' are located in a para relationship with respect to each other on Ar. In such a particular embodiment, E is 1.

[0030] In a specific embodiment, at least one X is located in an ortho relation or a para relation with respect to Y'.

[0031] In a specific embodiment, R attached to Ar b In addition to at least one R b represents Z'.

[0032] The present invention also has a chemical formula ( I' The invention relates to a composition (e.g., a pharmaceutical composition) comprising a compound of ) and a carrier (e.g., a pharmaceutically acceptable carrier).

[0033] In one aspect, the present invention relates to a chemical formula for use, for example, in therapy, as an imaging sensor, as a molecular switch, as a molecular machine, and / or a nanomachine ( I' The conjugate of ) and a composition comprising such a conjugate are provided.

[0034] In another aspect, the present invention further provides a chemical formula used in a method for delivering an activator to a cell ( I' The present invention provides a conjugate of ) and a pharmaceutical composition thereof, wherein the targeting moiety is selected to bind to a molecule associated with a target cell. In particular, the compound, conjugate, and composition inhibit abnormal cell growth or mammals ( for example It may be useful for treating proliferative disorders in humans, for example, the target cell is a cancer cell, and the targeting moiety is selected to bind to a molecule that is associated with cancer cells (and is not associated with healthy cells, or is at least preferentially associated with tumor cells rather than healthy cells).

[0035] Chemical formula ( I' The conjugate of ) and its pharmaceutical composition are mammals ( for example It may be useful for treating pathological conditions in humans, such as cancer, rheumatoid arthritis, multiple sclerosis, graft-versus-host disease (GVHD), transplant rejection, lupus, myositis, infections, immunodeficiency, such as AIDS, and inflammatory diseases. Brief explanation of the drawing

[0036] Figure 1 shows the results of the stability analysis of compound A-1. Figure 2 shows the results of the stability analysis of compound A-2. Figure 3 shows the results of the stability analysis of compound B-1. Figure 4 shows the results of the enzymatic cleavage assay of compound A-2. Figure 5 shows the results of the enzymatic cleavage assay of compound Int-T3. Figure 6 shows the results of in vitro analysis of conjugates T-DM1, T-2-AB, and T-3-AB against NCI-N87. Figure 7 shows the results of an in vitro analysis of conjugates T-DM1 and T-4-AB against NCI-N87. Figure 8 shows the results of an in vitro analysis of conjugates T-DM1, T-5-AB, and T-6-AB against SK-BR3. Specific details for implementing the invention

[0037] The present invention relates to compounds and conjugates comprising a cleavable linker and to uses thereof. Representative compounds and conjugates disclosed herein comprise an activator having a desired function or activity (e.g., chemical factor, biological factor, hormone, oligonucleotide, drug, toxin, ligand, probe for detection, etc.), a promoter that has undergone a chemical reaction (e.g., physicochemical reaction and / or biological reaction) under predetermined conditions, and an SO2 functional group located in relation to the promoter on an aryl or heteroaryl ring, wherein activating the promoter causes the conjugate to release the activator through 1,4-elimination, intramolecular cyclization, and 1,6-elimination. In some embodiments, the compounds and conjugates disclosed herein further comprise a targeting moiety having binding specificity to a desired target receptor or other molecule associated with a target cell ( for example, oligopeptide, polypeptide, antibody, etc. Includes ).

[0038] definition

[0039] The meaning of the term "alkyl" is understood in the art. For example, alone or, for instance, "alkoxy," "haloalkyl," "cycloalkyl," "heterocycloalkyl," etc. more "Alkyl," used as part of a large moiety, may refer to a fully saturated straight-chain or branched hydrocarbon. Typically, a straight-chain or branched alkyl group is an acyclic group having 1 to about 20 carbon atoms, preferably 1 to about 10 carbon atoms, unless otherwise defined. Examples of straight-chain and branched alkyl groups include, but are not limited to: methyl, ethyl, propyl, isopropyl, n -butyl, sec -butyl, tert -butyl, n -pentyl, n - Hexyl, n -Heptyl, and n -Octyl. C1-C6 straight-chain or branched alkyl groups are also referred to as "lower alkyl" groups. Alkyl groups with two open valencies are sometimes, as in alkylenes. " It is referred to by the suffix "ene". Exemplary alkylene groups include methylene, ethylene, propylene, etc.

[0040] Furthermore, the term “alkyl” (or “lower alkyl”) may include both “unsubstituted alkyl” and “substituted alkyl,” the latter referring to an alkyl moiety having a substituent that replaces a hydrogen on one or more carbons of a hydrocarbon backbone. Such substituents, if atypical, may include, for example, halogen, hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxyl, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidin, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclil, aralkyl, or aromatic or heteroaromatic moiety. Those skilled in the art will understand that the substituted moiety on the hydrocarbon chain may self-substitute if appropriate. For example, the substituents of the substituted alkyl may include alkyl, amino, azido, imino, amido, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamido, sulfamoyl and sulfonates), and silyl groups in substituted and unsubstituted forms, as well as ethers, alkylthio, carbonyl (including ketones, aldehydes, carboxylates, and esters), -CF3, -CN, etc. Exemplary substituted alkyls are listed below. The cycloalkyl may be further substituted with alkyl, alkenyl, alkoxy, alkylthio, aminoalkyl, carbonyl-substituted alkyl, -CF3, -CN, and other homologous groups.

[0041] Term "C x -C y"When used in combination with a chemical moiety, e.g., acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, it may comprise a group containing x to y carbons within the chain, where "x" and "y" are selected integers from 1 to about 20, and x is an integer of a value less than y, and x and y are not the same value. For example, the term "C x -C y "-alkyl" refers to a substituted or unsubstituted saturated hydrocarbon group, which is a haloalkyl group, e.g., trifluoromethyl and 2,2,2-trifluoroethyl, etc. Including, it comprises straight-chain alkyl and branched-chain alkyl groups containing x to y carbons within the chain. Terms "C2-C y -Alkenyl" and "C2-C y "-alkynyl" refers to a substituted or unsubstituted unsaturated aliphatic group that is similar in length and possible substitution to the aforementioned alkyl but contains at least one double or triple bond, respectively. As applied to heteroalkyls, "C x -C y" indicates that the group contains x to y number of carbons and heteroatoms in the chain. As applied to carbon-ring structures, e.g., aryl and cycloalkyl groups, "C x -C y " This indicates that the above ring contains x to y carbon atoms in the ring.

[0042] The meaning of the term "alkoxy" is understood in the art and may refer, for example, to an alkyl group having oxygen attached thereto, preferably a lower alkyl group. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert - Includes butoxy, etc.

[0043] The terms “hal,” “halo,” and “halogen” are used interchangeably throughout the specification and refer to fluorine or fluoro (F), chlorine or chloro (Cl), bromine or bromo (Br), or iodine or iodo (I).

[0044] The meaning of the term "cycloalkyl" is understood in the art and, for example, may refer to a fully saturated, substituted, or unsubstituted cyclic hydrocarbon. Cycloalkyls include monocyclic and bicyclic rings. Typically, a monocyclic cycloalkyl group has 3 to about 10 carbon atoms, more typically 3 to 8 carbon atoms, unless otherwise defined. The second ring of a bicyclic cycloalkyl may be selected from saturated, unsaturated, and aromatic rings. Cycloalkyls include bicyclic molecules in which 1, 2, or 3 or more atoms are shared between two rings. The term "fused cycloalkyl" refers to a bicyclic cycloalkyl in which each ring shares two adjacent atoms with the other ring. The second ring of a fused bicyclic cycloalkyl may be selected from saturated, unsaturated, and aromatic rings. A "cycloalkenyl" group is a cyclic hydrocarbon containing one or more double bonds.

[0045] The meaning of the term "aryl" is understood in the art and may refer, for example, to a substituted or unsubstituted single-ring aromatic group, wherein each atom of the ring is a carbon. Preferably, the ring is a 5- to 7-membered ring, more preferably a 6-membered ring. The term "aryl" also comprises a polycyclic ring system having two or more cyclic rings common to two adjacent rings, wherein at least one of the rings is aromatic, and for example, other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkyl, aryl, heteroaryl, and / or heterocyclyl. The aryl group includes benzene, naphthalene, phenanthrene, phenol, aniline, etc.

[0046] The meanings of the terms “heterocyclile” and “heterocycle” are understood in the art and may refer, for example, to a substituted or unsubstituted non-aromatic ring structure, preferably a 3- to 10-membered ring, more preferably a 3- to 7-membered ring, said ring structure comprising at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. Such a heterocycle also comprises a polycyclic ring system having two or more cyclic rings common to two adjacent rings at least two carbons, said rings at least one of which is heterocyclic, and for example , other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkyl, aryl, heteroaryl, and / or heterocyclyl. The heterocyclyl group includes, for example, piperidine, piperazine, pyrrolidine, morpholine, lactone, lactam, etc.

[0047] The meaning of the term "heteroaryl" is understood in the art and may refer, for example, to a substituted or unsubstituted aromatic monocyclic structure, preferably a 5- to 7-membered ring, more preferably a 5- to 6-membered ring, said ring structure comprising at least one heteroatom, preferably 1 to 4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heteroaryl" and "hetaaryl" also comprise a polycyclic ring system having two or more cyclic rings in which two or more carbons are common to two adjacent rings, said rings wherein at least one of said rings is heteroaromatic, for example, other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkyl, aryl, heteroaryl, and / or heterocyclyl. The heteroaryl group includes, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine.

[0048] The term "substituted" refers to a moiety having a substituent that replaces a hydrogen atom on one or more carbons or heteroatoms of the moiety. Those skilled in the art will understand that "substitution" or "substituted" includes the implied condition that such substitution is in accordance with the allowed valence of the substituted atom and the substituent, and that said substitution, for example It will be understood that this results in a stable compound that does not undergo spontaneous modifications such as rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" is considered to include all acceptable substituents of an organic compound.

[0049] In some embodiments, acceptable substituents include acyclic and cyclic, branched and unbranched, carbon-cyclic and heterocyclic, aromatic and non-aromatic substituents of the organic compound. There may be one or more acceptable substituents and may be the same or different for a suitable organic compound. For the present invention, a heteroatom, e.g., nitrogen, may have any acceptable substituent and / or hydrogen substituent of the organic compound described herein that satisfies the valence of the heteroatom. Substituents may include any substituents described herein, e.g., halogen, hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxyl, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidin, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclil, alkyl, aralkyl, or aromatic or heteroaromatic moiety. Those skilled in the art will understand that substituents may be substituted if appropriate. Unless specifically stated as "unsubstituted," designations of chemical moiety herein are understood to include substituted variants. For example, references to an "aryl" group or moiety implicitly include both substituted and unsubstituted variants.

[0050] The term "subject" to which administration is being considered is, for example, human ( in other words , male or female of any age group, for example , pediatric subjects ( for example , infants, children, adolescents) or adult subjects ( for example , young adult, middle-aged adult, or elderly)) and / or other primates ( for example, cynomolgus monkey, rhesus monkey); commercially related mammals, such as cattle, pigs, horses, sheep, goats, cats, and / or dogs; and / or commercially related birds, such as chickens, ducks, geese, quail, and / or turkeys. A preferred subject is a human.

[0051] As used herein, a therapeutic agent that “prevents” a disorder or condition may refer to a compound that, for example, reduces the occurrence of a disorder or condition in a treated sample compared to an untreated control sample in a statistical sample, or reduces the severity of one or more symptoms of a disorder or condition or delays the onset thereof compared to an untreated control sample.

[0052] The term "treating" includes prophylactic and / or therapeutic treatment. The term "prophylactic or therapeutic" treatment is technically recognized and includes administration to one or more hosts of the target compositions. Clinical signs of an unwanted pathological condition ( for example If administered prior to a disease or other unwanted condition in the host animal, the treatment is prophylactic and ( in other words , protects the host against the development of unwanted pathologies), on the other hand, if administered after signs of an unwanted pathology, the treatment is therapeutic, ( in other words , intended to alleviate, improve, or stabilize existing unwanted pathological conditions or their side effects).

[0053] In certain embodiments, the compounds and conjugates disclosed herein may be used alone or may be administered co-administered with another type of therapeutic compound or formulation. As used herein, the phrase “co-administered” refers to any form of administration of two or more different therapeutic compounds such that a second compound is administered while the previously administered therapeutic compound is still effective in the body ( for example, two compounds are simultaneously effective in a subject, and this may include a synergistic effect of the two compounds). For example, different therapeutic compounds and conjugates may be administered simultaneously or sequentially, in the same formulation or in separate formulations. In certain embodiments, different therapeutic compounds and conjugates may be administered within 1 hour, 12 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, or one week from each other. Thus, a subject receiving such treatment may benefit from the combined effect of different therapeutic compounds and conjugates.

[0054] The terms “abnormal cell growth” and “proliferative disorder” are used interchangeably herein. “Abnormal cell growth” as used herein refers to cell growth independent of normal regulatory mechanisms, unless otherwise indicated ( for example , loss of contact inhibition). This includes, for example, the following abnormal growths: (1) tumor cells (tumors) that proliferate by expressing a mutated tyrosine kinase or by overexpression of a receptor tyrosine kinase; (2) benign and malignant cells of other proliferative diseases in which abnormal tyrosine kinase activation occurs; (3) any tumor proliferated by receptor tyrosine kinase; (4) any tumor proliferated by abnormal serine / threonine kinase activation; and (5) benign and malignant cells of other proliferative diseases in which abnormal serine / threonine kinase activation occurs.

[0055] The terms “cancer” and “cancerous” refer to or describe physiological pathological conditions in mammals typically characterized by uncontrolled cell growth. “Tumor” contains one or more cancerous cells. Examples of cancer include, but are not limited to, the following: carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancy. A more specific example of such cancer is squamous cell carcinoma ( for example, squamous cell carcinoma), small cell lung cancer, non-small cell lung cancer ("NSCLC"), lung cancer including adenocarcinoma of the lung and squamous cell carcinoma of the lung, cancer of the peritoneum, hepatocellular carcinoma, gastric or stomach cancer including gastric cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, liver tumor, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, liver carcinoma, anal carcinoma, penile carcinoma, acute leukemia, as well as head / brain and neck cancer.

[0056] Compounds and conjugates of the present invention

[0057] Chemical formula ( I' A conjugate of ) or a pharmaceutically acceptable salt thereof is provided herein:

[0058]

[0059] Here:

[0060] CB is a targeting moiety;

[0061] cb and n are each independently integers having a value of 1 to about 20, preferably 1 to about 10;

[0062] Each DL independently has the chemical formula ( I" It is a device having the structure of ):

[0063]

[0064] Each Q is an activator independently connected to L' by a heteroatom, preferably O or N;

[0065] Z' is the chemical formula independently in each case ( I" The structure of ) (CB) cbA linker connecting to, a solubilizing group, a reactive group (e.g., a precursor group), a solid surface (e.g., a particle), a stabilizing group, a chelator, a biopolymer (e.g., an immunoglobulin, nucleic acid, protein, oligopeptide, polypeptide, antibody, fragment or repeat of an antigenic polypeptide), an activator, or a detectable moiety, provided that at least one occurrence of Z' is of the chemical formula ( I" The structure of ) (CB) cb Connect to;

[0066] Each L' is a spacer moiety independently attached to SO2 through a heteroatom selected from O, S, and N, preferably O or N, and the cleavage of the bond between L' and SO2 is selected to facilitate the cleavage of the bond between L' and Q to release an activator;

[0067] Each X is independently -O-, -C(R b )(R c )-, or -N(R c )-, preferably -C(R b )(R c )-and;

[0068] E is an integer having a value of 1, 2, or 3, preferably 1;

[0069] Ar is a 6-membered aryl or 6-membered heteroaryl ring;

[0070] Y' is -N(R a )-, -O-, or -S- and;

[0071] At least one X is located in an ortho relation or a para relation with respect to Y' of Ar;

[0072] TG is an inducing group that generates N, O, or S atoms capable of initiating the emission of SO2 and (Q)q-(L')w when cleaved;

[0073] Each q is an integer having a value of 1 to about 20, preferably 1 to about 10, independently;

[0074] Each w and x is an integer that independently has a value of 0 or 1;

[0075] Each R a and R c is independently hydrogen or lower alkyl; and

[0076] At least one R b Each R under the condition that is Z' b is independently Z', hydrogen, or lower alkyl; or

[0077] R b and R c ..., together with the atoms to which they are attached, form a 3-5-membered ring, preferably a 3-4-membered ring;

[0078] However, when w is 0, q is 1.

[0079] In a specific embodiment, each X is located in an ortho relation or a para relation with respect to Y' of Ar.

[0080] In a specific embodiment, at least one X and Y' are ortho-related to each other in Ar. In a specific such embodiment, E is 2 or 3. In a specific embodiment where E is 2, two occurrences of X are located ortho-related to Y'.

[0081] In another embodiment, at least one X and Y' are located in a para relationship with each other in Ar. In a specific such embodiment, E is 1.

[0082] In a specific embodiment, at least one X is located in an ortho relation or a para relation with respect to Y'.

[0083] In a specific preferred embodiment, R attached to Ar b At least one R that is not b represents Z'.

[0084] Each activator may be any suitable activator as described in more detail below. While many traditional conjugation methods require having specific functional groups, such as amine or hydroxyl groups, to form a stable linker, the disclosure herein provides a strategy for forming a link using functional groups, such as phenol and tertiary amines, to form a stable linker in the conjugate disclosed herein, while still allowing release under predetermined conditions that activate the inducing group. When the inducing group is activated, SO2 is released and (Q) q -(L') w While releasing the -H moiety, the aryl / heteroaryl unit undergoes an elimination reaction (e.g., 1,6-elimination of the group at the para position), where H is bonded to the heteroatom of Q or L' that was previously bonded to the SO2 moiety. For the group in the ortho relationship, the activity of the inducing group can initiate similar 1,4-elimination and / or intramolecular cyclization, thus (Q) q -(L') w -H is released. Examples of cyclization triggered by 1,6-elimination and 1,4-elimination and / or cleavage of the inducing group are shown in Schematics 1 and 2.

[0085] Many suitable initiators are known in the art, and exemplary initiators and the conditions for activating them are discussed below, as with the moiety described for Y below. Some initiators contain N, O, or S atoms but are in a non-nucleophilic form. For example, the NO2 group is an initiator that is reduced to an NH2 or NHOH group capable of reacting with SO2 under reducing conditions and initiates a elimination reaction. The acetate group is an initiator that is hydrolyzed to a hydroxyl group capable of inducing 1,6-elimination of a self-sacrificial moiety under hydrolysis conditions. Other initiators do not contain N, O, or S atoms but are converted to nucleophilic N, O, or S atoms when activated. For example, the boronate group is an initiator that is converted to a hydroxyl group capable of reacting with SO2 under oxidizing conditions (e.g., peroxide). Preferably, the inducing group is selected such that the conditions activating it do so selectively without cleavage or disintegration of other parts of the conjugate, e.g., the targeting moiety. In some embodiments, once a nucleophilic N, O, or S atom is generated, that atom attacks the SO2 moiety within the molecule to form a ring, while the moiety (Q) q -(L') w - It emits H (where H is bonded to the heteroatom of Q or L' previously attached to the SO2 moiety).

[0086] In an embodiment where w is 0, q is 1 and Q is directly attached to SO2 through a heteroatom. Thus, the induced group activation attacks the SO2 moiety within the molecule to form a ring, generating a nucleophilic heteroatom that releases the activator QH (where H is bonded to the heteroatom previously attached to SO2).

[0087] In an embodiment where w is 1, L' may be selected to allow the attachment of multiple occurrences of Q, which may be identical or different. Thus, each instance of Q is attached indirectly to SO2 through a spacer moiety. In such an embodiment, induced group activation attacks the SO2 moiety within the molecule to form a ring, while the moiety (Q) q A nucleophilic heteroatom is generated that releases -L'-H (where H is bonded to the heteroatom at L' previously attached to SO2). In such an embodiment, the released heteroatom triggers an intramolecular reaction that releases activator(s) Q (e.g., if Q has a tertiary amine attached to L' as a quaternary ammonium) or QH. For example, the heteroatom may undergo intramolecular cyclization with an ester moiety formed by the hydroxyl of QH, ring formation, and release of activator QH. Alternatively, the heteroatom may undergo intramolecular tautomerization that releases activator Q or QH.

[0088] Ar can be any suitable ring, including a bicyclic or other polycyclic ring, such that the moiety undergoing intramolecular cyclization is maintained in very close proximity to promote the reaction after activation of the initiating group. Although other types of rings, e.g., cycloalkenyl or heterocycloalkenyl, may force similar geometric structures, planar characteristics of aromatic and heteroaromatic rings are preferred, as the rigid geometry of substituents on such rings ensures good arrangement of the reaction moiety. 5- or 6-membered rings, and / or the number or identity of heteroatoms in the ring, and / or substituents on other rings (e.g., electron-donating or electron-withdrawing substituents) may be selected to control the rate of cyclization based on the bond angle obtained in the ring. Similarly, more flexible forms of cycloalkyl and heterocyclil rings may be useful when slowing down the rate of intramolecular cyclization is required.

[0089] Z' can be any suitable linker connecting Ar to one or more CB groups. Typically, the linker is a moiety, such as a polyethylene glycol (PEG) moiety, a peptide sequence, or a charge-retaining moiety (e.g., carboxylate, amine, nitrogen-containing ring, etc.) to balance the hydrophobic characteristics of any alkyl chain that may be included. etc. ), etc. By including it, it must be sufficiently hydrophilic to frustrate the aggregation of the conjugate and to promote its water solubilization (solubilization group). Since it is often advantageous to manufacture conjugates in a modular manner, Z' may contain a linking unit, which is a functional group resulting from the conjugation of one reaction moiety to another. Representative linking units are described in more detail below ( for example , discussed in relation to variable Z, and typically the linking group is amide, triazole, oxime, carbamate, etc. It includes. A representative Z' group is L 1' -Z groups are included as discussed in more detail below. In some embodiments, all DL groups attached to each CB are identical, but in other embodiments, each CB may be attached to two or more distinct DL groups. For example, some DL groups may have inducers activated under a first condition, and other DL groups may have inducers activated under a second condition, so that, for example, one activator may be selectively released under the first condition, but the second activator may be selectively released under the second condition.

[0090] The present disclosure also includes, chemical formula ( I" As described in ). provided a compound that can be used as a formation intermediate or reagent of the group DL in chemical formula (I'). Accordingly, in some embodiments, chemical formula ( IaA compound of ) or a pharmaceutically acceptable salt thereof is provided herein:

[0091]

[0092] Here:

[0093] Each Q is, independently, an activator connected to L' by a heteroatom, preferably O or N;

[0094] Z' is independently a member and a connector in each case;

[0095] Each L' is a linker independently attached to SO2 through a heteroatom selected from O, S, and N, preferably O or N, and the cleavage of the bond between L' and SO2 is selected to facilitate the cleavage of the bond between L' and Q to release an activator;

[0096] Each X is independently -O-, -C(R b )(R c )-, or -N(R c )-, preferably -C(R b )(R c )-and;

[0097] E is an integer having a value of 1, 2, or 3, preferably 1;

[0098] Ar is a 6-membered aryl or 6-membered heteroaryl ring;

[0099] Y' is -N(R a )-, -O-, or -S- and;

[0100] At least one X is located in an ortho or para relation to Y' on Ar;

[0101] TG is an inducing group that generates N, O, or S atoms capable of initiating the emission of SO2 and (Q)q-(L')w when cleaved;

[0102] Each q is an integer having a value of 1 to about 20, preferably 1 to about 10, independently;

[0103] Each of w and x is an integer that independently has a value of 0 or 1;

[0104] Each R a and R c is independently hydrogen or lower alkyl; and

[0105] Each R b is at least one R b is independently Z', hydrogen, or lower alkyl, provided that it is Z; or

[0106] R b and R c They form a 3-5-membered ring, preferably a 3-4-membered ring, together with the carbon atoms to which they are attached;

[0107] However, when w is 0, q is 1.

[0108] In some embodiments, the chemical formula herein ( Ia ) A compound of or a pharmaceutically acceptable salt thereof is provided:

[0109]

[0110] Here:

[0111] Each Q is an activator independently connected to L' by a heteroatom, preferably O or N;

[0112] Z' is a reactive group (e.g., a precursor group that can be used to attach a compound to a triggering agent, such as CB);

[0113] Each L' is a linker independently attached to SO2 through a heteroatom selected from O, S, and N, preferably O or N, and the cleavage of the bond between L' and SO2 is selected to facilitate the cleavage of the bond between L' and Q to release an activator;

[0114] Each X is independently -O-, -C(R b )(R c )-, or -N(Rc )-, preferably -C(R b )(R c )-and;

[0115] E is an integer having a value of 1, 2, or 3, preferably 1;

[0116] Ar is a 6-membered aryl or 6-membered heteroaryl ring;

[0117] Y' is -N(R a )-, -O-, or -S- and;

[0118] At least one X is located in an ortho or para relation to Y' on Ar;

[0119] TG is an inducing group that generates N, O, or S atoms capable of initiating the emission of SO2 and (Q)q-(L')w when cleaved;

[0120] Each q is an integer having a value of 1 to about 20, preferably 1 to about 10, independently;

[0121] Each of w and x is an integer that independently has a value of 0 or 1;

[0122] Each R a and R c is independently hydrogen or lower alkyl; and

[0123] Each R b is at least one R b is independently Z', hydrogen, or lower alkyl, provided that it is Z; or

[0124] R b and R c They form a 3-5-membered ring, preferably a 3-4-membered ring, together with the carbon atoms to which they are attached;

[0125] However, when w is 0, q is 1.

[0126] In a specific embodiment, each X is located in an ortho-relative or para-relative to Y' on Ar.

[0127] In a specific embodiment, at least one X and Y' are located in an ortho relationship with respect to each other on Ar. In a specific such embodiment, E is 2 or 3. In a specific embodiment where E is 2, two occurrences of X are located in an ortho relationship with respect to Y'.

[0128] In another embodiment, at least one X and Y' are located in a para relationship with respect to each other on Ar. In a specific such embodiment, E is 1.

[0129] In a specific embodiment, at least one X is located in an ortho relationship or a para relationship to Y'.

[0130] In a specific embodiment, R attached to Ar b Excluding at least one R b represents Z'.

[0131] Chemical formula ( I' ) and ( Ia In a specific implementation of ), -Y' is -N(R a )-, -O-, or -S-. In some such embodiments, TG is β-galactoside, β-glucuronide, or a combination of β-galactoside and β-glucuronide.

[0132] Chemical formula ( I' ) and ( Ia In some implementations of ), (L') w is an activator that connects each Q to SO2; and each Q is connected to one of the L' groups through a heteroatom, preferably O or N, and forms -O-, -OC(O)-, -OC(O)O- or -OC(O)NH- linkings including the heteroatom of Q.

[0133] In another implementation, (Q) q -(L') w - is selected from the following:

[0134] and

[0135]

[0136] Here:

[0137] Q is an activator connected to L' through a heteroatom, preferably O or N;

[0138] X 4 is absent or forms -O-, -OC(O)-, -OC(O)O-, or -OC(O)NH- linkages containing a heteroatom of Q;

[0139] X 1 -O- or -NR a -, preferably -O- and;

[0140] X 2 is -O-, -OC(O)-, -OC(O)O- or -OC(O)NH- and;

[0141] X 3 is -OC(=O)- and;

[0142] w' is an integer with a value of 1, 2, 3, 4, or 5;

[0143] R 9 and R 10 Each is independently hydrogen, alkyl, aryl, or heteroaryl, wherein the alkyl, aryl, and heteroaryl are unsubstituted or for example , alkyl, -(CH2) u NH2, -(CH2) u NR u1 R u2 , and -(CH2) u SO2R u3 Substituted with one or more substituents selected from;

[0144] R u1 , R u2 , and R u3 Each is independently hydrogen, alkyl, aryl, or heteroaryl; and

[0145] u is an integer with a value from 1 to about 10.

[0146] In some such implementations, (Q) q -(L') w - is selected from the following:

[0147]

[0148] In addition, the present invention has a chemical formula ( I' Conjugate or chemical formula according to ) Ia Provides an intermediate for preparing a compound according to ), wherein (Q) among the above formulas q -(L') w is replaced by a leaving group, e.g., a halogen (preferably fluorine), (Q) q -(L') w Allows attachment of.

[0149] In certain such embodiments, Z' comprises a reactive group (e.g., a precursor group as discussed in more detail below for Z), which induces the compound, e.g., CB ( for example , chemical formula discussed in more detail above ( I To manufacture the compound of '), hard surface ( for example It can be used to form beads, nanoparticles, solid-supported arrays, or sensor particles, or to attach to any other molecule or support of interest.

[0150] In certain embodiments, Z' comprises a reactive group (e.g., a precursor group as discussed in more detail below for Z) that can be used to attach a compound to a targeting moiety such as CB.

[0151] In a specific preferred embodiment, the compound of formula (I') is selected from the following:

[0152]

[0153]

[0154]

[0155] and

[0156]

[0157] Here:

[0158] X is -O-, -C(R b )(R c )-, or -N(R c )-and;

[0159] R 1 is a C1-C6 alkyl; and

[0160] R 21 and R 22 Each is independently hydrogen or C1-C6-alkyl.

[0161] R 9 and R 10 Each is independently hydrogen, alkyl, aryl, or heteroaryl, where the alkyl, aryl, and heteroaryl are unsubstituted or, for example, alkyl, -(CH2) u NH2, -(CH2) u NR u1 R u2 , and -(CH2) u SO2R u3 It is replaced with one or more substituents selected from.

[0162] R u1 , R u2 , and R u3 Each is independently hydrogen, alkyl, aryl, or heteroaryl; and

[0163] u is an integer with a value from 1 to about 10.

[0164] In another embodiment, chemical formula ( I' The compound of ) is selected from the following:

[0165]

[0166]

[0167] and

[0168] In another embodiment, chemical formula ( I' The compound of ) is selected from the following:

[0169]

[0170]

[0171] and

[0172]

[0173] In another embodiment, chemical formula ( I' The compound of ) is selected from the following:

[0174]

[0175]

[0176]

[0177] and

[0178]

[0179] In another embodiment, chemical formula ( I' The compound of ) is selected from the following:

[0180]

[0181]

[0182] In another embodiment, chemical formula ( I' The compound of ) is selected from the following:

[0183] and

[0184]

[0185] In a specific preferred embodiment, Z is the chemical formula ( F ), ( G ), ( H ), ( J ), ( K ), ( L ), ( M ), or ( N It is a connector having the structure of ):

[0186]

[0187] Here:

[0188] * is the attachment point for CB;

[0189] ** is the attachment point for Ar;

[0190] R e is alkyl and;

[0191] X" is -O-, -S-, -NH-, or -CH2-;

[0192] X 4 -NHC(O)-(CH2) g -NH- or -C(O)NH-(CH2) h -NH- and;

[0193] W b1 and W b2 Each is independently -C(O)NH-, -NHC(O)-, or And;

[0194] L 2 is optionally a spacer moiety, and one or more substituents, e.g., C1-C6 alkyl, C5-C 14 It may be further substituted with aryl, and C3-C8 heteroaryl, and said alkyl, aryl, and heteroaryl for example , C1-C 10 Alkyl, -(CH2) u NH2, -(CH2) u NR u1 R u2 , -(CH2) u CO2H, -(CH2) u CO2R u1 , and -(CH2) u SO2R u3 It may be further substituted with one or more substituents selected from the group consisting of, and R u1 , R u2 , and R u3 Each independently hydrogen, C1-C 15 Alkyl, C6-C 20 Aryl or C3-C 10 It is heteroaryl; and u is an integer having a value from 1 to about 10;

[0195] R 12is hydrogen, C1-C8 alkyl, or an amino acid moiety, e.g., a natural amino acid moiety;

[0196] b, c, d, e, g, h, o, and qq are each independently integers having a value from 1 to about 10; and

[0197] s' is an integer with a value from 1 to about 10.

[0198] In another embodiment, Z is the chemical formula ( F' ), ( G' ), ( H' ), ( J' ), ( K' ), ( L' ), ( M' ), or ( N' It is a connector having the structure of ):

[0199] s

[0200]

[0201]

[0202] In a specific preferred embodiment, CB is selected from the following:

[0203] and

[0204]

[0205] In a specific preferred embodiment, (Q) q -(L') w is selected starting from the following:

[0206]

[0207]

[0208]

[0209] ,

[0210] Here, * is -SO2- (Q) q -(L') w - indicates the point where it is attached.

[0211] In some embodiments, (Q) q -(L') w is selected from the following.

[0212]

[0213]

[0214] Here, * is -SO2- (Q) q -(L') w - indicates the point where it is attached.

[0215] Chemical formula ( I A compound of ) or a pharmaceutically acceptable salt thereof is also provided herein:

[0216]

[0217] Here:

[0218] Each X is independently -O-, -CH2-, or -NR'-;

[0219] E is an integer having a value of 1, 2, or 3, preferably 1;

[0220] R' is hydrogen, C1-C6-alkyl, C6-C 14 -aryl, or C2-C 20 -Heteroaryl;

[0221] Ar is C5-C 20 - Aromatic ring, C2-C 20 -Heteroaromatic ring, C2-C 30 - Fusion ring, or C5-C 20 -Aromatic ring-C2-C 20 -Heteroaromatic ring;

[0222] R is Ar or -L 1' -Z-(CB) cb , preferably -L 1' -Z-(CB) cb It is a substituent of the upper part;

[0223] L 1' is C1-C 200 -alkylene or C1-C 200It is an alkylene, wherein it further comprises at least one of a peptide bond, an amino bond, an ether bond, a triazole bond, a tetrazole bond, a sugar bond, a sulfonamide bond, a phosphonate bond, a sulfo bond, or a dendrimer structure;

[0224] Z is CB and L 1' or reactive group ( for example , this is a connecting unit that connects precursor groups capable of connecting to CB;

[0225] CB is a targeting moiety, such as a ligand, having the characteristic of binding to a receptor;

[0226] cb is an integer with a value of 0, 1, or 2;

[0227] n is an integer with a value of 1, 2, 3, or 4;

[0228] Y is -NO2, -OC(O)(CH2) r C(O)R 1 , -O(CH2) r -Ar 1 -NO2, -NHOH, -NHNH2, -BR 2 R 3 , or -Y'-TG, preferably Y is -NO2, -OC(O)(CH2) r C(O)R 1 , -O(CH2) r -Ar 1 -NO2, -NHNH2, -BR 2 R 3 , or -Y'-TG and;

[0229] R 1 is a C1-C6 alkyl;

[0230] r is an integer with a value of 1, 2, 3, 4, or 5;

[0231] Ar 1 is C6-C 20 Arilen;

[0232] R 2 and R 3Each is independently hydrogen, C1-C6-alkyl, C1-C6-alkoxy, or hydroxy;

[0233] R a , R b , R c , and R d Each is independently hydrogen or C1-C6 alkyl;

[0234] Y' is -(CH2) x NR"-, -(CH2) x O-, or -(CH2) x S-go;

[0235] R" is hydrogen or C1-C6 alkyl;

[0236] x is an integer with a value of 0 or 1;

[0237] TG is an inducing agent;

[0238] Q is -Q 1 or -L'-(Q 1 ) w And;

[0239] L' has -O- or -NR"'- at one end and -O-, -OC(O)-, -O(CO)O-, -OC(O)NR"'- or -OC(O)NR at the other end 4 C7-C with CH2O- 30 - It is a hydrocarbon spacer, and the above -O-, -OC(O)-, -O(CO)O- or -OC(O)NR""- is C7-C 30 It may be additionally included within the hydrocarbon spacer, and the above C7-C 30 Hydrocarbon spacers are one or more substituents, e.g., C1-C6 alkyl, C5-C 14 aryl, and further substituted with C3-C8 heteroaryl, wherein the alkyl, aryl, and heteroaryl are, for example , C1-C 10 Alkyl, -(CH2) u NH2, -(CH2) u NR u1 R u2 , -(CH2) u CO2H, -(CH2)u CO2R u1 , and -(CH2) u SO2R u3 It may be further substituted with one or more substituents selected from the group consisting of, and R u1 , R u2 , and R u3 Each independently hydrogen, C1-C 15 Alkyl, C6-C 20 Aryl or C3-C 10 It is heteroaryl; and u is an integer having a value from 1 to about 10;

[0240] Q 1 -OH, -NH-, -NR 5 R 6 An activator comprising at least one functional group selected from -SH, -SO2NH2, or -COOH;

[0241] R 4 is hydrogen, C1-C6-alkyl, C5-C 14 -aryl, or C3-C8-heteroaryl, wherein the alkyl, aryl, and heteroaryl are substituted or unsubstituted;

[0242] R 5 and R 6 Each is independently hydrogen, C1-C6-alkyl, C3-C9-cycloalkyl, or C5-C 10 - Heteroaryl, wherein the heteroaryl is substituted or unsubstituted;

[0243] R"' and R"' each are independently hydrogen or C1-C6-alkyl; and

[0244] w is an integer with a value of 1, 2, 3, 4, or 5.

[0245] In some embodiments, chemical formula ( I Compounds of ) functional groups capable of inducing intramolecular cyclization by external stimuli ( for example Includes , Y). In a specific embodiment, the functional group is introduced at the ortho-position and / or para-position for each X.

[0246] In some embodiments, R' is C1-C6-alkyl, C6-C 14 -aryl, or C2-C 20 - It is a heteroaryl.

[0247] In some embodiments, Ar is C5-C 20 - Aromatic ring, C2-C 20 -Heteroaromatic ring, C2-C 30 - Fusion ring, or C5-C 20 -Aromatic ring-C2-C 20 - It is a heteroaromatic ring. For example, Ar can be a benzene ring, a naphthalene ring, a pyridine ring, or a quinolone ring. Preferably, Ar is a benzene ring or a naphthalene ring. In some embodiments, the chemical formula ( I) The compound of has the chemical formula ( II )to It is a compound having a different structure or a pharmaceutically acceptable salt thereof:

[0248]

[0249] In another embodiment, chemical formula ( I The compound of ) has the chemical formula ( III It is a compound having a structure according to ) or a pharmaceutically acceptable salt thereof:

[0250]

[0251] In another embodiment, chemical formula ( I The compound of ) has the chemical formula ( IV It is a compound having a structure according to ) or a pharmaceutically acceptable salt thereof:

[0252]

[0253] In another embodiment, chemical formula ( I The compound of ) has the chemical formula ( V It is a compound having a structure according to ) or a pharmaceutically acceptable salt thereof:

[0254]

[0255] In another embodiment, chemical formula (I The compound of ) has the chemical formula ( VI It is a compound having a structure according to ) or a pharmaceutically acceptable salt thereof:

[0256]

[0257] In another embodiment, chemical formula ( I The compound of ) has the chemical formula ( VII It is a compound having a structure according to ) or a pharmaceutically acceptable salt thereof:

[0258]

[0259] In another embodiment, chemical formula ( I The compound of ) has the chemical formula ( VIII It is a compound having a structure according to ) or a pharmaceutically acceptable salt thereof:

[0260]

[0261] In another embodiment, chemical formula ( I The compound of ) has the chemical formula ( IX It is a compound having a structure according to ):

[0262]

[0263] In some embodiments, the compound has the chemical formula ( I ), ( II ), ( III ), ( IV ), ( V ), ( VI ), ( VII ), ( VIII ), or ( IXA compound of which R is hydrogen, halogen (hal), aldehyde, acetal, ketal, -R*, -OR*, -SR*, -NR*R**, -C(hal)3, -CN, -OCN, -SCN, -N=C=O, -NCS, -NO, -NO2, -N3, -NC, -C(O)R*, -OC(O)R*, -OS(O)R*, -S(O)2R*, -S(O)2OR*, -OS(O)OR*, -OS(O)2OR*, -S(O)NR*R**, -S(O)2NR*R**, -S(O)R*, -OP(O)(OR*)2, -P(O)(OR*)2, -OP(OR*)2, -OP(OR*)N(R**)2, -OP(O)(OR*)N(R**)2, -PR*, Selected from -P(O)2, -P(O)R*, -C(O)hal, -C(S)R*, -CO2R*, -C(S)OR*, -C(O)SR*, -C(S)SR*, -C(O)NR*R**, -C(S)NR*R**, -C(=NR*)NR*R**, -NR*C(O)R**, -NR*S(O)2OR**, -NR*S(O)R**, -NR*C(O)NR**, -SS-R*, or -R*SSR**, where: R* and R** are each independently hydrogen, C1-C 18 Alkyl, C6-C 20 Aryl, C3-C 15 Heterocycle or C3-C 20 It is a heteroaryl.

[0264] In some embodiments, the compound has the chemical formula ( I ), ( II ), ( III ), ( IV ), ( V ), ( VI ), ( VII ), ( VIII ), or ( IX A compound of ), wherein R is hydrogen or *-(L a -A1-L b -L c -Z) m -CB and;

[0265] Here:

[0266] La is a single bond or C1-C 20 - It is an alkylene;

[0267] A 1 is -C(O)NR*-, -NR*C(O)-, -NR*-, -O-, -PO3-, -OPO3-, -SO-, -SO2- or -SO3- and;

[0268] L b -(CH2CH2O) a - or -(CH2) a - and;

[0269] R* is hydrogen, C1-C 18 -alkyl, C6-C 20 -Aryl, C3-C 15 - Complex call, or C3-C 20 It is heteroaryl;

[0270] a is an integer with a value from 1 to about 20;

[0271] L c is a single bond or C1-C 20 - It is an alkylene;

[0272] n is an integer with a value of 1 or 2; and

[0273] Z is CB and L c It is a connection unit that connects; or

[0274] Z is isocyanide, isothiocyanide, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, alkene, halide, tosylate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivative (-C(O)C≡CR aIt is a precursor selected from ), and R a is C1-C 10 -alkyl), and dihydrogen phosphate (-OP(=O)(OH)2);

[0275] CB is a ligand capable of binding to a targeting moiety, such as a receptor; and

[0276] m is an integer with a value of 0, 1, or 2.

[0277] In some embodiments, the compound has the chemical formula ( I ), ( II ), ( III ), ( IV ), ( V ), ( VI ), ( VII ), ( VIII ), or ( IX A compound of ), wherein R is hydrogen or *-L a -A1-L b -L c -Z and; here:

[0278] L a is a single bond or C1-C 20 - It is an alkylene;

[0279] A 1 is -C(O)NR*-, -NR*C(O)-, -NR*-, -O-, -PO3-, -PO4-, -SO-, -SO2-, or -SO3-;

[0280] L b -(CH2CH2O) a - or -(CH2) a - and;

[0281] R* is hydrogen, C1-C 18 -alkyl, C6-C 20 -Aryl, C3-C 15 -Heterocycle, or C3-C 20 -Heteroaryl;

[0282] a is an integer with a value from 1 to about 20;

[0283] L cis a single bond or C1-C 20 - It is an alkylene;

[0284] n is an integer with a value of 1 or 2; and

[0285] Z is isocyanide, isothiocyanide, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, alkene, halide, tosylate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivative (-C(O)C≡CR a It is a precursor selected from, and R a is C1-C 10 -alkyl), and dihydrogen phosphate (-OP(=O)(OH)2);

[0286] In some embodiments, the compound has the chemical formula ( I ), ( II ), ( III ), ( IV ), ( V ), ( VI ), ( VII ), ( VIII ), or ( IX A compound of ), where R is *(-L a -A1-L b -L c -Z) m -CB and; here:

[0287] L a is a single bond or C1-C 20 - It is an alkylene;

[0288] A 1is -C(O)NR*-, -NR*C(O)-, -NR*-, -O-, -PO3-, -PO4-, -SO-, -SO2- or -SO3-;

[0289] L b -(CH2CH2O) a - or -(CH2) a - and;

[0290] R* is hydrogen, C1-C 18 -alkyl, C6-C 20 -Aryl, C3-C 15 -Heterocycle, or C3-C 20 -Heteroaryl;

[0291] a is an integer with a value from 1 to about 20;

[0292] L c is a single bond or C1-C 20 - It is an alkylene;

[0293] n is an integer with a value of 1 or 2;

[0294] Z is CB and L c It is a connection unit that connects;

[0295] CB is a ligand having the characteristic of binding to a targeting moiety, e.g., a receptor; and

[0296] m is an integer with a value of 1 to 2.

[0297] In some embodiments, the compound has the chemical formula ( I ), ( II ), ( III ), ( IV ), ( V ), ( VI ), ( VII ), ( VIII ), or ( IX It is a compound of ), wherein L' is C7-C 30 It is a hydrocarbon spacer, wherein it additionally includes -O-, -OC(O)-, -O(CO)O- or -OC(O)NR""-.

[0298] In some embodiments, the compound has the chemical formula (I ), ( II ), ( III ), ( IV ), ( V ), ( VI ), ( VII ), ( VIII ), or ( IX A compound of ), wherein Q is selected from -L'-(Q 1 ) w am:

[0299]

[0300] Here:

[0301] Q 1 -OH, -NR 5 R 6 An activator comprising at least one functional group selected from -SH, and -COOH;

[0302] Q 2 is -NR 5 R 6 It is an active agent containing;

[0303] X 1 is -O- or -NR"'- and;

[0304] X 2 and X 4 Each is independently absent or selected from -O-, -OC(O)-, -OC(O)O-, and -OC(O)NH-;

[0305] X 3 is -OC(=O)- and;

[0306] R 5 and R 6 is identical to the previously defined one;

[0307] R 9 and R 10 Each is independently hydrogen, C1-C6 alkyl, C6-C 14 It is an aryl or C3-C9 heteroaryl, and R9 and R 10 The alkyl, aryl, and heteroaryls are C1-C 10 Alkyl, -(CH2) u NH2, -(CH2) uNR u1 R u2 , and -(CH2) u SO2R u3 It may be further substituted with one or more substituents selected from the group consisting of, and R u1 , R u2 , and R u3 Each independently hydrogen, C1-C 15 Alkyl, C6-C 20 Aryl or C3-C 10 It is heteroaryl; and u is an integer having a value from 1 to about 10;

[0308] R"' is hydrogen or C1-C6-alkyl; and

[0309] w is an integer with a value of 1, 2, 3, 4, or 5.

[0310] In a specific implementation, -L'-(Q 1 ) w is selected from the following:

[0311]

[0312] -OH, -NR 5 R 6 Q, Q selected from , -SH, and -COOH 1 , or Q 2 At least one functional group serves as a linkage site for the activator on L'. The functional group may exist as an ester, thioester, carbonate, carbamate, amide, sulfonamide, sulfonate, sulfate, or as part of another suitable linkage; namely, -OH, -NR 5 R 6 The -SH and -COOH moieties do not exist as they are, but the activator is part of the conjugate.

[0313] In some embodiments, Q 2 is -NR 5 R 6 It is an activator comprising, said activator can be bonded in a quaternary amine structure, for example, among said activator -NR 5R 6 The moiety can form a quaternary amine link having L'.

[0314] Chemical formula ( I" ), ( Ia ), ( I ), ( II ), ( III ), ( IV ), ( V ), ( VI ), ( VII ), ( VIII ), or ( IX In some implementations of ), R 4 is a substituted alkyl, aryl, or heteroaryl. In some such embodiments, R 4 is C1-C 10 Alkyl, -(CH2) u NH2, -(CH2) u NR u1 R u2 , -(CH2) u CO2H, -(CH2) u CO2R u1 , and -(CH2) u SO2R u3 Substituted with one or more substituents selected from, and R u1 , R u2 , and R u3 Each independently hydrogen, C1-C 15 -alkyl, C6-C 20 -aryl, or C3-C 10 - It is a heteroaryl; and u is an integer with a value from 1 to about 10.

[0315] Chemical formula ( I" ), ( Ia ), ( I ), ( II ), ( III ), ( IV ), ( V ), ( VI ), ( VII ), ( VIII ), or ( IX In some implementations of ), R 5 and / or R 6is -NR 7 R 8 It is a heteroaryl substituted with R 7 and R 8 Each is independently hydrogen, C1-C6-alkyl, C3-C9-cycloalkyl, or C5-C 14 - It is Aril.

[0316] Chemical formula ( I" ), ( Ia ), ( I ), ( II ), ( III ), ( IV ), ( V ), ( VI ), ( VII ), ( VIII ), or ( IX In some implementations of ), Q or -(L') w -(Q) q is selected starting from the following:

[0317]

[0318]

[0319] and ,

[0320] Here, * is Q or (Q) in -SO2- q -(L') w - indicates the attachment point.

[0321] Chemical formula ( I" ), ( Ia ), ( I ), ( II ), ( III ), ( IV ), ( V ), ( VI ), ( VII ), ( VIII ), or ( IX In some implementations of ), Q or -(L') w -(Q) q Selected from the following:

[0322]

[0323]

[0324] Here, * is Q or (Q) in -SO2- q -(L') w - indicates the attachment point.

[0325] In a specific embodiment, chemical formula ( I ), ( II ), ( III ), ( IV ), ( V ), ( VI ), ( VII ), ( VIII ), or ( IX A compound of ) is provided in this specification, wherein:

[0326] Y is -NO2, -OC(O)(CH2) r C(O)R 1 , -O(CH2) r -Ar 1 -NO2, -NHOH, -BR 2 R 3 or -Y'-TG, preferably Y is -NO2, -OC(O)(CH2) r C(O)R 1 , -O(CH2) r -Ar 1 -NO2, -BR 2 R 3 or -Y'-TG and;

[0327] R 1 is a C1-C6 alkyl;

[0328] r is an integer with a value of 1, 2, 3, 4, or 5;

[0329] Ar 1 is phenylene, biphenylene, or naphthylene;

[0330] R 2 and R 3 Each is independently hydrogen, C1-C6-alkyl, C1-C6-alkoxy, or hydroxy;

[0331] Y' is -(CH2) x NR"-, -(CH2) xO- or -(CH2) x S-go;

[0332] R" is hydrogen or C1-C6-alkyl;

[0333] x is an integer with a value of 0 or 1;

[0334] R" is hydrogen or C1-C6-alkyl; and

[0335] TG is an inducing group, such as β-galactoside, β-glucuronide, or a combination of β-galactoside and β-glucuronide.

[0336] In a specific implementation example, the formula ( I ), ( II ), ( III ), ( IV ), ( V ), ( VI ), ( VII ), ( VIII ), or ( IX The compound of ) is selected from the following:

[0337]

[0338]

[0339]

[0340]

[0341]

[0342]

[0343]

[0344] Here:

[0345] R 1 is a C1-C6 alkyl;

[0346] R 21 and R 22 Each is independently hydrogen or acetyl;

[0347] R is hydrogen, *-L a -A1-L b -L c -Z, or chemical formula (F ), ( G ), ( H ), ( J ), ( K ), ( L ), ( M ), or ( N It is a device having the structure of ):

[0348]

[0349] L a is a single bond or C1-C 20 It is an alkylene;

[0350] A 1 is -C(O)NH-, -NHC(O)-, -NH-, -O-, -PO3-, -PO4-, -SO-, -SO2- or -SO3-;

[0351] L b -(CH2CH2O) a - or -(CH2) a - and;

[0352] a is an integer with a value from 1 to about 20;

[0353] L c is C1-C 20 It is an alkylene;

[0354] X" is -O-, -S-, -NH-, or -CH2-;

[0355] W b1 and W b2 Each is independently -C(O)NH-, -NHC(O)-, or And;

[0356] R 12 is hydrogen, C1-C8 alkyl, amino acid moiety, -(CH2) s COR 13 , or -(CH2) p NR 14 R 15 And;

[0357] R 13 is OH or -NH(CH2) s' (X"CH2CH2) s"It is Z;

[0358] R 14 and R 15 Each independently hydrogen or -(C(O)(CH2) s' (X"CH2CH2) s" Z) m -CB and;

[0359] X" is -O-, -S-, -NH-, or -CH2-;

[0360] R e is a C1-C8-alkyl or -(L 1' -Z) m -CB and;

[0361] X 4 -NHC(O)-(CH2) g -NH- or -C(O)NH-(CH2) h -NH- and;

[0362] b, c, d, e, g, h, o, and q are each independently integers having a value from 1 to about 10;

[0363] p is an integer having a value from 1 to about 10;

[0364] s and s" are each independently integers having a value from 0 to about 10;

[0365] s' is an integer having a value from 1 to about 10;

[0366] m is an integer with a value of 0 or 1;

[0367] Z is isocyanide, isothiocyanide, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, alkene, halide, tosylate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10-Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, Carboxylic acid (-COOH), Acetylene (-C≡CH), Azide (-N3), Amino (-NH2), Sulfonic acid (-SO3H), Alkynone derivative (-C(O)C≡CR a and R a is C1-C 10 alkyl), or dihydrogen phosphate (-OP(=O)(OH)2);

[0368] CB is a ligand selected from the following:

[0369] and

[0370] and

[0371] Q is selected from the following:

[0372]

[0373]

[0374]

[0375] and Here, * indicates the attachment site of Q to -SO2-.

[0376] In some embodiments, chemical formula ( I ), ( II ), ( III ) ( IV ), ( V ), ( VI ), ( VII ), ( VIII ), or ( IX The compound of ) is selected from the following:

[0377]

[0378]

[0379]

[0380]

[0381]

[0382]

[0383]

[0384] Here:

[0385] R 1 is a C1-C6 alkyl;

[0386] R 21 and R 22 Each is independently hydrogen or acetyl;

[0387] R is hydrogen, *-L a -A1-L b -L c -Z, or chemical formula ( F ), ( G ), ( H ), ( J ), ( K ), ( L ), ( M ), or ( N It is a structure of ):

[0388]

[0389] L a is a single bond or C1-C 20 It is an alkylene;

[0390] A 1 is -C(O)NH-, -NHC(O)-, -NH-, -O-, -PO3-, -PO4-, -SO-, -SO2- or -SO3-;

[0391] L b -(CH2CH2O) a - or -(CH2) a - and;

[0392] a is an integer with a value from 1 to about 20;

[0393] L c is C1-C 20 It is an alkylene;

[0394] X" is -O-, -S-, -NH-, or -CH2-;

[0395] W b1 and W b2Each is independently -C(O)NH-, -NHC(O)-, or And;

[0396] R 12 is hydrogen, C1-C8 alkyl, amino acid moiety, -(CH2) s COR 13 , or -(CH2) p NR 14 R 15 And;

[0397] R 13 is OH or -NH(CH2) s' (X"CH2CH2) s" It is Z;

[0398] R 14 and R 15 Each independently hydrogen or -(C(O)(CH2) s' (X"CH2CH2) s" Z) m -CB and;

[0399] X" is -O-, -S-, -NH-, or -CH2-;

[0400] R e is a C1-C8-alkyl or -(L 1' -Z) m -CB and;

[0401] X 4 -NHC(O)-(CH2) g -NH- or -C(O)NH-(CH2) h -NH- and;

[0402] b, c, d, e, g, h, o, and q are each independently integers having a value from 1 to about 10;

[0403] p is an integer having a value from 1 to about 10;

[0404] s and s" are each independently integers having a value from 0 to about 10;

[0405] s' is an integer having a value from 1 to about 10;

[0406] m is an integer with a value of 0 or 1;

[0407] Z is isocyanide, isothiocyanide, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, alkene, halide, tosylate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 -Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, Carboxylic acid (-COOH), Acetylene (-C≡CH), Azide (-N3), Amino (-NH2), Sulfonic acid (-SO3H), Alkynone derivative (-C(O)C≡CR a and R a is C1-C 10 alkyl), or dihydrogen phosphate (-OP(=O)(OH)2);

[0408] CB is a ligand selected from the following:

[0409] and

[0410] and

[0411] Q is selected from the following:

[0412]

[0413] Here, * is

[0414] It indicates the attachment point of Q to -SO2-.

[0415] Release of active agent

[0416] As described above, in certain embodiments, the compound and conjugate disclosed herein undergo an intramolecular cyclization reaction following a chemical reaction that activates the inducing group (Q, Q 1 , Q 2One or more activators (as presented by) can be dissociated. In certain embodiments, the chemical reaction is a physicochemical reaction and / or a biochemical reaction.

[0417] In some embodiments, the compound and conjugate disclosed herein are X ( for example It includes a nucleophilic functional group (Y or Y') introduced from an adjacent atom on Ar for (, -CH2-). Typically, the nucleophilic functional group is masked by a promoter (TG) as further detailed below. Upon activation, the promoter releases the nucleophilic functional group to react with an adjacent SO2 moiety in intramolecular cyclization and / or 1,4-elimination, ultimately involving one or more activators (Q, Q 1 , or Q 2 ) or releases their protonated forms. In some such embodiments, one or more activators are released via an intramolecular cyclization reaction after a chemical reaction, a physicochemical reaction and / or a biochemical reaction (see, e.g., Scheme 1), or the activators are released via 1,6-elimination or 1,4-elimination after an intramolecular cyclization reaction (see, e.g., Scheme 2).

[0418] For example, the mechanism in which Y is -Y'-TG is illustrated in Reaction Scheme 1:

[0419] Reaction Equation 1:

[0420]

[0421]

[0422] Q is The mechanism in this case is illustrated in Reaction Scheme 2:

[0423] Reaction Equation 2:

[0424]

[0425]

[0426] The mechanism of the multi-substituted structure is illustrated in Reaction Scheme 3.

[0427]

[0428] In some embodiments, Q 1 is an activator comprising at least one functional group selected from -OH, -NH-, -SH, and -COOH when released. According to these embodiments, as further described herein, Q 1 is conjugated to compounds as described herein by -OH, -NH-, -SH, and -COOH through functional groups selected from, for example, esters, amides, thioesters, carbamates, ureas, oximes, hydrazones, etc. In some such embodiments, Q 2 is Q 1 Used instead, and Q 2 is an amine group-containing drug. In another embodiment, Q 2 is an activator capable of binding to ammonium units. In another embodiment, Q 2 is Q 2 Upon release, it can dissociate into its initial form having an amine group, and the activator may be a drug, a toxin, an affinity ligand, a probe for detection, or a combination thereof.

[0429] In some embodiments, the compounds and conjugates disclosed herein are chemically and physiologically stable. In some such embodiments, the compounds and conjugates disclosed herein selectively release a drug by reaching a desired target cell with minimal dissociation of the activator in the blood.

[0430] Inducing agent (TG)

[0431] In some embodiments, the conjugate of the present invention comprises a causative group (TG). The TG is a group that can be cleaved by a chemical reaction, e.g., a biological reaction, preferably optionally cleaved. Generally, the causative group serves to block the nucleophilic nature of the Y or Y' group, thereby thereby stabilizing ( for exampleThe compounds and conjugates disclosed herein are provided with (by preventing self-sacrificial or intramolecular cyclization before the conjugate reaches a target site or experiences a predetermined trigger condition). Upon activation, the trigger group releases a nucleophilic Y or Y' group and causes self-sacrificial or intramolecular cyclization to occur as described above.

[0432] In some embodiments, TG is a sequence (e.g., a peptide sequence, For example, Dipeptides (Val-Cit, Val-Ala) or moiety comprising TEV, trypsin, thrombin, cathepsin B, cathepsin D, cathepsin K, caspase 1, matrix metalloproteinases (MMPs), and enzymes ( for example , oxidoreductase, transferase, hydrolase, degrading enzyme, isomerase, ligase, etc. Can be hydrolyzed by ) and / or phosphodiesters, phospholipids, esters, β-galactose, β-glucose, fructose, oligosaccharides, and other homologous substances It is recognized by a homogeneous one that may include a moiety selected from.

[0433] In some embodiments, TG comprises a reactive chemical moiety or functional group, wherein it is a nucleophilic reagent condition ( for example It can be cleaved under silyl ether, 2-N-acyl nitrobenzenesulfonamide, unsaturated vinyl sulfide, sulfonamide after activation, malon dialdehyde-indole derivative, levulinoy ester, hydrazone, or acyl hydrazone.

[0434] In some embodiments, TG may include a reactive chemical moiety or functional group, provided that it is under basic reagent conditions ( for example It can be cleaved under , 2-cyanoethyl ester, ethylene glycol disuccinate, 2-sulfonylethyl ester, alkyl thioester, or thiophenyl ester.

[0435] In some embodiments, TG may include a reactive chemical moiety or functional group that can be cleaved by light irradiation ( for example , 2-nitrobenzyl derivatives, phenacyl esters, 8-quinolinylbenzenesulfonate, coumarin, phosphotriesters, bis-arylhydrazone, or bi-thiophionic acid derivatives).

[0436] In some embodiments, TG may include a reactive chemical moiety or functional group, which may be cleaved under reducing agent conditions ( for example , hydroxylamine, disulfide, revelinate, nitro, or 4-nitrobenzyl derivative).

[0437] In some embodiments, TG may include a reactive chemical moiety or functional group, which may be cleaved under acidic conditions ( for example , sugars, tert-butylcarbamate analogs, dialkyl or diaryl dialkoxysilanes, orthoesters, acetals, aconityl, hydrazone, β-thiopropionate, phosphoramidate, imines, trityl, vinyl ethers, polyketals, and alkyl 2-(diphenylphosphino)benzoate derivatives; alkyl esters, 8-hydroxyquinoline esters, and picolinate esters).

[0438] In some embodiments, TG may include a reactive chemical moiety or functional group, provided that it is subject to oxidation conditions ( for example , boronate, proximity diol, paramethoxybenzyl derivative, or selenium compound) It can be cut off under.

[0439] In a specific preferred embodiment, TG comprises a sugar, which can be cleaved under acidic or enzymatic conditions. In a specific preferred embodiment, the initiating group is -NO2, which can be cleaved under reducing conditions. In a specific preferred embodiment, the initiating group is boronate, which can be cleaved under oxidizing conditions. In a specific preferred embodiment, the initiating group is an ester, which can be cleaved under acidic, basic, or enzymatic conditions. In a specific preferred embodiment, the initiating group is a hydrazone, which can be cleaved under nucleophilic or acidic conditions. In a specific preferred embodiment, the initiating group is hydroxylamine, which can be cleaved under reducing conditions.

[0440] Sugar-inducing agent

[0441] In some embodiments, the compounds and conjugates disclosed herein comprise a sugar-inducing group, for example, a group selected from the following:

[0442] and

[0443] Here , each R 21 is independently hydrogen or OR 21 hydroxy protector ( for example , selected to be acetyl); and R 22 is hydrogen or lower alkyl (for example, C1-C6-alkyl) In certain embodiments, the hydroxy protecting group may be used in organic synthesis and includes, but is not limited to: methyl ether, methoxymethyl ether, methylthiomethyl ether, 2-methoxyethoxymethyl ether, bis(2-chloroethoxy)methyl ether, tetrahydropyranyl ether, tetrahydrothiopyranyl ether, 4-methoxytetrahydropyranyl ether, 4-methoxytetrahydrothiopyranyl ether, tetrahydrofuranyl ether, 1-ethoxyethyl ether, 1-methyl-1-methoxyethyl ether, 2-(phenylselenyl)ethyl ether, t-butyl ether, allyl ether, benzyl ether, o-nitrobenzyl ether, triphenyl methyl ether, α-naphthyldiphenyl methyl ether, p-Methoxyphenyldiphenylmethyl ether, 9-(9-phenyl-10-oxo)anthyl ether, trimethylsilyl ether, isopropyldimethylsilyl ether, t-butyldimethylsilyl ether, t-butyldiphenylsilyl ether, tribenzylsilyl ether, triisopropylsilyl ether, formate ester, acetate ester, trichloroacetate ester, phenoxyacetate ester, isobutyrate ester, pivaloate ester, adamanthoate ester, benzoate ester, 2,4,6-trimethylbenzoate ester, methyl carbonate, 2,2,2-trichloroethyl carbonate, allyl carbonate, p-nitrophenyl carbonate, benzyl carbonate, p-nitrobenzyl carbonate, S-benzylthiocarbonate, N-phenylcarbamate, nitrate ester, 2,4-dinitrophenylsulfenate ester, etc. However, it is not limited to these.

[0444] Protector as an inducing agent

[0445] In some embodiments, TG is a group that can be cleaved by a chemical reaction, a physicochemical reaction, and / or a biological reaction. In certain embodiments, TG is a protecting group. In some such embodiments, the protecting group is an amine group protecting group, an alcohol protecting group, or a thiol protecting group.

[0446] Amine protector

[0447] In certain embodiments, the amine protecting group is a general protecting group that can be used in organic synthesis and includes, but is not limited to: m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methyl carbamate, alkyl carbamate, 9-fluorenylmethyl carbamate, 2,2,2-trichloroethyl carbamate, 2-trimethylsilylethyl carbamate (Teoc), t-butyl carbamate (Boc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate (Ipaoc), 8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, benzyl carbamate, p-methoxybenzyl carbamate, p-nitrobenzyl carbamate, diphenyl methyl carbamate, acetamide, chloroacetamide, trichloroacetamide, Phenylacetamide, Benzamide, N-Phthalimide, N-2,3-Diphenylmaleimide, N-2,5-Dimethylpyrrole, N-1,1-Dimethylthiomethyleneamine, N-Benzylideneamine, Benzenesulfenamide, o-Nitrobenzenesulfenamide, Triphenylmethylsulfenamide, p-Toluenesulfonamide, Methanesulfonamide, etc. However, it is not limited to these.

[0448] alcohol protector

[0449] In certain embodiments, the alcohol protecting group is a general protecting group that can be used in organic synthesis and is: methyl ether, methoxymethyl ether (MOM ether), benzyloxymethyl ether (BOM ether), 2-(trimethylsilyl)ethoxymethyl ether (SEM ether), phenylthiomethyl ether (PTM ether), 2,2-dichloro-1,1-difluoroethyl ether, p-bromophenacyl ether, chloropropylmethyl ether, isopropyl ether, cyclohexyl ether, 4-methoxybenzyl, 2,6-dichlorobenzyl ether, 4-(dimethylaminocarbonyl)benzyl ether, 9-anthrylmethyl ether, 4-picolyl ether, methylthiomethyl ether (MTM ether), 2-methoxyethoxymethyl ether (MEM ether), Bis(2-chloroethoxy)methyl ether, tetrahydropyranyl ether (THP ether), tetrahydrothiopyranyl ether, 4-methoxytetrahydropyranyl ether, 4-methoxytetrahydrothiopyranyl ether, tetrahydrofuranyl ether, 1-ethoxyethyl ether, 1-methyl-1-methoxyethyl ether, 2-(phenylselenyl)ethyl ether), t-butyl ether, allyl ether, benzyl ether, o-nitrobenzyl ether, triphenylmethyl ether, α-naphthyldiphenylmethyl ether, p-methoxyphenyldiphenylmethyl ether, 9-(9-phenyl-10-oxo)anthyl ether, trimethylsilyl ether (TMS ether), isopropyldimethylsilyl ether, t-butyldimethylsilyl ether (TBDMS ether), t-butyldiphenylsilyl ether, tribenzylsilyl ether, triisopropylsilyl ether, formate ester, acetate ester, trichloroacetate ester, phenoxyacetate ester, isobutyrate ester, pivaloate ester, adamanthoate ester, benzoate ester, 2,4,6-trimethylbenzoate (methitoate) ester, methyl carbonate, 2,2,2-trichloroethyl carbonate, allyl carbonate, p-nitrophenyl carbonate, benzyl carbonate, p-nitrobenzyl carbonate, S-benzyl thiocarbonate, N-phenylcarbamate, nitrate ester, 2,Includes, but is not limited to, 4-dinitrophenylsulfenate esters, dimethylphosphinyl esters (DMP esters), dimethylthiophosphinyl esters (MPT esters), aryl methanesulfonates, aryl toluenesulfonates, etc.

[0450] Thiol protector

[0451] In certain embodiments, thiol protecting groups may be used in organic synthesis and include, but are not limited to: S-benzyl thioether, Sp-methoxybenzyl thioether, So- or p-hydroxyl or acetoxybenzyl thioether, Sp-nitrobenzyl thioether, S-4-picolyl thioether, S-2-picolyl N-oxide thioether, S-9-anthylmethyl thioether, S-9-fluorenylmethyl thioether, S-methoxymethyl monothioacetal, A-acetyl derivative, S-benzoyl derivative, S-(N-ethyl carbamate), S-(N-methoxymethyl carbamate), etc.

[0452] Connector

[0453] In some embodiments, the compounds and conjugates disclosed herein include a linking group connecting each CB and Ar through a covalent bond. A typical linking group is a stable, non-hydrolyzable moiety, e.g., C 10 -C 100 It is a linear or branched, saturated or unsaturated alkylene. In a specific embodiment, the linking unit satisfies at least two of the following four criteria, and more preferably at least three:

[0454] (i) At least one -CH2- in the alkylene moiety is substituted (i.e. replaced by) one or more heteroatoms selected from -NH-, -C(=O), -O-, -S- and -P-;

[0455] (ii) At least one heteroarylene is contained within an alkylene moiety;

[0456] (iii) at least one amino acid moiety, sugar bond, peptide bond, or amide bond is contained within an alkylene moiety; and

[0457] (iv) The above alkylene is C1-C 20 Alkyl, C6-C 20 Aryl C1-C8 alkyl, -(CH2) s COOH, and -(CH2) p It may be further substituted with one or more substituents selected from the group consisting of NH2, wherein s is an integer having a value from 0 to 10, and p is an integer having a value from 1 to about 10.

[0458] In a specific embodiment, the connection unit comprises at least two of the following, and more preferably at least three:

[0459] (i) at least one heteroatom selected from -NH-, -C(=O), -O-, -S- and -P-;

[0460] (ii) at least one heteroarylene;

[0461] (iii) at least one amino acid moiety, sugar bond, peptide bond, or amide bond; and

[0462] (iv) The above alkylene is C1-C 20 Alkyl, C6-C 20 Aryl C1-C8 alkyl, -(CH2) s COOH, and -(CH2) p It may be further substituted with one or more substituents selected from the group consisting of NH2, wherein s is an integer having a value from 0 to 10, and p is an integer having a value from 1 to about 10.

[0463] In another embodiment, the linker connecting each CB and Ar includes a functional group generated through a click chemical reaction.

[0464] In an alternative embodiment, the linking unit includes a reaction functional group capable of participating in a click chemical reaction.

[0465] Click chemistry is a reaction that can be carried out under mild conditions and involves a functional group not typically found in biological molecules ( for example It is extremely selective for azide groups, acetylene groups, etc. Therefore, such reactions can be carried out in the presence of complex inducing groups, targeting moiety, etc. Furthermore, click chemistry possesses high reaction specificity. For example, the click chemical reaction between an azide group and an acetylene group proceeds selectively without interference from other functional groups present in the molecule. For example, azide-acetylene click chemistry can yield triazole moiety in high yield.

[0466] Therefore, in some embodiments, the connector connecting each CB and Ar is or Includes, where V is a single bond, -O-, -S-, -NR 21 -, -C(O)NR 22 -, -NR 23 C(O)-, -NR 24 SO2-, or -SO2NR 25 -It could be, R 21 to R 25 Each is independently hydrogen, (C1-C6)alkyl, (C1-C6)alkyl(C6-C 20 )aryl, or (C1-C6)alkyl(C3-C 20 ) It may be a heteroaryl, r may be an integer having a value from 1 to about 10, p may be an integer having a value from 0 to about 10, q may be an integer having a value from 1 to about 10, and L" may be a single bond.

[0467] In another embodiment, the connection unit connecting each CB and Ar is Equation ( A Indicated as ) Is It is a connector:

[0468]

[0469] Here:

[0470] * is the attachment point for CB;

[0471] ** is the attachment point for Ar;

[0472] W a1 , W a2 , and W a3 Each independently -NH-, -C(=O)-, or (-CH2-) b And;

[0473] W b1 is an amide bond or triazolilene;

[0474] P 1 is W a3 and Y 2 It is a linker connecting, and is an amino acid moiety, a peptide bond, or an amide bond;

[0475] L c is an alkylene;

[0476] Y 2 is a single bond, -W a4 -(CH2) c -W b2 -(CH2) d -W a5 -, or -W a6 -(CH2) e -CR e R f -X- and;

[0477] R e is a C1-C8 alkyl or CB-W a7 -Y3-W c1 -(CH2) f - and;

[0478] R f is BW a7 -Y 3 -W c1 -(CH2) f - and;

[0479] X is -NHC(=O)-(CH2) g -W a8 - or -C(=O)NH-(CH2) h -W a9 - and;

[0480] W a4 , W a5 , W a6 , W a7 , W a8 , and W a9 Each is independently -NH-, -C(=O)-, or -CH2-;

[0481] W b2 is an amide bond or triazolilene;

[0482] W c1 is -NHC(=O)- or -C(=O)NH- and;

[0483] Y 3 -(CH2) i -(X'CH2CH2) j -(CH2) k - and;

[0484] X' is -O-, -S-, -NH-, or -CH2-;

[0485] CB is the same as defined above;

[0486] b, c, d, e, f, g, h, i, and j are each independent integers having a value from 1 to about 10;

[0487] k and y are each independently integers having a value from 0 to about 10;

[0488] Y 1 -(CH2) q -(CH2CH2X") o - or -(CH2) q -(X"CH2CH2) o - and;

[0489] X" is -O-, -S-, -NH-, or -CH2-; and

[0490] o and q are integers with values ​​ranging from 1 to about 10.

[0491] In some embodiments, P 1 is the formula ( B ) or ( C Includes at least one unit indicated by ):

[0492]

[0493] Here:

[0494] R 12 is hydrogen, C1-C8-alkyl, amino acid side chain, e.g., natural amino acid side chain ( for example , H, methyl, isopropyl, isobutyl, sec-butyl, S-methyl thioether, benzyl, indole, pyrrolidine, pyrroline, hydroxymethyl, tyrosyl, lysyl, imidazole, glycyl, glutamyl, carbamoylbutanoic acid, carboxamide, aspartic acid, 1-hydroxyethyl, and 2-hydroxyethyl), -(CH2) s COR 13 or -(CH2) p NR 14 R 15 And;

[0495] R 13 is OH or -NH(CH2) s' (X"CH2CH2) s" It is Z;

[0496] R 14 and R 15 Each independently hydrogen or -(C(O)(CH2) s' (X"CH2CH2) s" Z) m -CB and;

[0497] X" is -O-, -S-, -NH-, or -CH2-;

[0498] Z and CB are the same as defined above;

[0499] p is an integer having a value from 1 to about 10;

[0500] s and s" are integers having values ​​from 0 to about 10;

[0501] s' is an integer with a value from 1 to about 10; and

[0502] m is an integer with a value of 0 or 1.

[0503] In some embodiments of chemical formula (B) or (C):

[0504] R 12 is hydrogen, alkyl, amino acid side chain, -(CH2) s C(O)R 13 or -(CH2) p NR 14 R 15 And;

[0505] p is an integer having a value from 1 to about 10;

[0506] s is an integer having a value from 0 to about 10;

[0507] R 13 is OH or -NH(CH2) s' (X"'CH2CH2) s" Z"-(CB) m And;

[0508] R 14 and R 15 Each independently hydrogen or -C(O)(CH2) s' (X"'CH2CH2) s" Z"-(CB) m And;

[0509] s" is an integer with a value from 0 to about 10;

[0510] s' is an integer having a value from 1 to about 10;

[0511] m is an integer with a value of 0 or 1;

[0512] X"' is -O-, -S-, -NH-, or -CH2-; and

[0513] Z" is CB R 14 or R 15 It is a linker that connects to the rest of; or Z" is a linker containing a reactive group.

[0514] In a part of such an embodiment of chemical formula (B) or (C):

[0515] R 13 is OH or -NH(CH2) s' (X"'CH2CH2)s" Z" and;

[0516] R 14 and R 15 Each independently hydrogen or -C(O)(CH2) s' (X"'CH2CH2) s" Z" and; and

[0517] Z" is isocyanide, isothiocyanide, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, alkene, halide, tosylate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivative (-C(O)C≡CR a It is a reaction precursor of a linked unit selected from, where R a is C1-C 10 -alkyl), and dihydrogen phosphate (-OP(=O)(OH)2).

[0518] In other such embodiments of chemical formula (B) or (C):

[0519] R 13 is OH or -NH(CH2) s' (X"'CH2CH2) s" Z"CB and;

[0520] R 14 and R 15 Each independently hydrogen or -C(O)(CH2) s' (X"'CH2CH2) s" Z"CB and; and

[0521] Z" is CB, and R formed from a precursor selected from the following. 14 or R 15It is a linking unit that connects to the remainder: isocyanide, isothiocyanide, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, alkene, halide, tosylate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivative (-C(O)C≡CR a , R a is C1-C 10 -alkyl), and dihydrogen phosphate (-OP(=O)(OH)2).

[0522] In some implementations, Y 2 is selected from a single combination or the following:

[0523] and

[0524]

[0525] Here:

[0526] W b2 -C(O)NH-, -NHC(O)-, or And;

[0527] R e is a C1-C8-alkyl or -(L 1' -Z-) m It is CB;

[0528] R f is BW b2' -(CH2) i -(X"CH2CH2) j -NH-C(=O)-(CH2) f - and;

[0529] X a -NHC(=O)-(CH2) g-NH- or -C(O)NH-(CH2) h -NH- and;

[0530] W b2' is -C(O)NH- or -NHC(=O)- and;

[0531] Each of c, d, e, f, g, h, i, and j is an integer having a value from 1 to about 10 independently;

[0532] X" is -O-, -S-, -NH-, or -CH2-; and

[0533] L 1' , Z, m, and B are equal as defined above.

[0534] In a specific embodiment, the connecting unit connecting each CB and Ar is (CH2) connected to each other by a covalent bond b , L c , (P 1 ) a , W a1 , W a2 , W a3 , Y 1 , and Y 2 It is a connecting unit including a unit, and here:

[0535] W a1 , W a2 , and W a3 Each is independently -NH-, -C(O)-, or -CH2-;

[0536] W b1 is an amide bond or triazolilene;

[0537] P 1 is an amide bond, an amino acid residue, or a peptide;

[0538] L c is an alkylene;

[0539] Y 1 -(CH2) q -(CH2CH2X") o - or -(CH2) q -(X"CH2CH2X") o - and;

[0540] X" is -O-, -S-, -NH- or -CH2- and;

[0541] Y 2 is a single combination or a group selected from the following:

[0542]

[0543] and

[0544] W b2 is an amide bond or triazolilene;

[0545] a is 0 to 10;

[0546] b, c, and d are each independently integers having a value from 1 to about 10; and

[0547] o and q are each integers with values ​​ranging from 1 to about 10 independently.

[0548] In some implementations, R 12 is a natural amino acid side chain. In another embodiment, R 12 is a non-natural amino acid side chain.

[0549] In some embodiments, the connecting unit connecting each CB and Ar is the chemical formula ( A) Table It is a connecting device that is being used:

[0550]

[0551] Here:

[0552] * is the attachment point for CB; and

[0553] ** is the attachment point for Ar.

[0554] In some such implementations, P 1 Is

[0555] or Igo

[0556] Here:

[0557] R 12is hydrogen, alkyl, amino acid side chain, -(CH2) s COOH or -(CH2) p NH2 and;

[0558] p is an integer with a value from 1 to about 10; and

[0559] s and s" are each integers that independently have a value from 0 to about 10.

[0560] In some implementations, P 1 Is

[0561] or Igo

[0562] Here:

[0563] R 12 is hydrogen, alkyl, amino acid side chain, -(CH2) s C(O)R 13 or -(CH2) p NR 14 R 15 And;

[0564] p is an integer having a value from 1 to about 10;

[0565] s is an integer having a value from 0 to about 10;

[0566] R 13 is OH or -NH(CH2) s' (X"'CH2CH2) s" Z"-(CB) m And;

[0567] R 14 and R 15 Each independently hydrogen or -C(O)(CH2) s' (X"'CH2CH2) s" Z"-(CB) m And;

[0568] s" is an integer with a value from 0 to about 10;

[0569] s' is an integer having a value from 1 to about 10;

[0570] m is an integer with a value of 0 or 1;

[0571] X"' is -O-, -S-, -NH-, or -CH2-; and

[0572] Z" is CB R 14 or R 15 It is a linker that connects to the rest of; or Z" is a linker containing a reactive group.

[0573] P 1 In some of such implementation examples:

[0574] R 13 is OH or -NH(CH2) s' (X"'CH2CH2) s" Z" and;

[0575] R 14 and R 15 Each independently hydrogen or -C(O)(CH2) s' (X"'CH2CH2) s" Z" and; and

[0576] Z" is isocyanide, isothiocyanide, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, alkene, halide, tosylate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivative (-C(O)C≡CR a It is a reaction precursor of a linked unit selected from, where R a is C1-C 10 -alkyl), and dihydrogen phosphate (-OP(=O)(OH)2).

[0577] P 1 In other such implementations of:

[0578] R13 is OH or -NH(CH2) s' (X"'CH2CH2) s" Z"CB and;

[0579] R 14 and R 15 Each independently hydrogen or -C(O)(CH2) s' (X"'CH2CH2) s" Z"CB and; and

[0580] Z" is CB, R formed from a precursor selected from the following. 14 or R 15 It is a linking unit that connects to the remainder: isocyanide, isothiocyanide, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, alkene, halide, tosylate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivative (-C(O)C≡CR a , R a is C1-C 10 -alkyl), and dihydrogen phosphate (-OP(=O)(OH)2).

[0581] In an alternative embodiment, the linking unit connecting CB and Ar is of the chemical formula ( F ), ( G ), ( H ), ( J ), ( K ), ( L ), ( M ), or ( N It is a connector indicated by ) and:

[0582]

[0583] Here:

[0584] R e is alkyl and;

[0585] X 4 -NHC(O)-(CH2) g -NH- or -C(O)NH-(CH2) h -NH- and;

[0586] e, g, and h are each independently integers having a value from 1 to about 10; and

[0587] s' is an integer with a value from 1 to about 10.

[0588] ceremony ( F ), ( G ), ( H ), ( I ), ( J ), ( K ), ( L ), or ( M In some implementation examples of ):

[0589] R 12 is hydrogen, alkyl, amino acid side chain, -(CH2) s C(O)R 13 or -(CH2) p NR 14 R 15 And;

[0590] p is an integer having a value from 1 to about 10;

[0591] s is an integer having a value from 0 to about 10;

[0592] R 13 is OH or -NH(CH2) s' (X"'CH2CH2) s" Z"-(CB) m And;

[0593] R 14 and R 15 Each independently hydrogen or -C(O)(CH2) s' (X"'CH2CH2) s" Z"-(CB) m And;

[0594] s" is an integer with a value from 0 to about 10;

[0595] s' is an integer having a value from 1 to about 10;

[0596] m is an integer with a value of 0 or 1;

[0597] X"' is -O-, -S-, -NH-, or -CH2-; and

[0598] Z" is CB R 14 or R 15 It is a linker that connects to the rest of; or Z" is a linker containing a reactive group.

[0599] Chemical formula ( F ), ( G ), ( H ), ( I ), ( J ), ( K ), ( L ), or ( M In a part of such an implementation example of ):

[0600] R 13 is OH or -NH(CH2) s' (X"'CH2CH2) s" Z" and;

[0601] R 14 and R 15 Each independently hydrogen or -C(O)(CH2) s' (X"'CH2CH2) s" Z" and; and

[0602] Z" is isocyanide, isothiocyanide, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, alkene, halide, tosylate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivative (-C(O)C≡CR a It is a reaction precursor of a linked unit selected from, where R a is C1-C 10 -alkyl), and dihydrogen phosphate (-OP(=O)(OH)2).

[0603] Chemical formula ( F ), ( G ), ( H ), ( I ), ( J ), ( K ), ( L ), or ( M In a part of such an implementation example of ):

[0604] R 13 is OH or -NH(CH2) s' (X"'CH2CH2) s" Z"CB and;

[0605] R 14 and R 15 Each independently hydrogen or -C(O)(CH2) s' (X"'CH2CH2) s" Z"CB and; and

[0606] Z" is CB, R formed from a precursor selected from the following. 14 or R 15 It is a linking unit that connects to the remainder: isocyanide, isothiocyanide, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, alkene, halide, tosylate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10Cycloalkynyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynone derivative (-C(O)C≡CR a , R a is C1-C 10 -alkyl), and dihydrogen phosphate (-OP(=O)(OH)2).

[0607] Targeting Moity

[0608] The compounds and conjugates of the present invention may additionally comprise a ligand or targeting moiety, CB. In some embodiments, the ligand or targeting moiety is any molecular recognition element, which may undergo specific interactions with at least one other molecule through, for example, non-covalent bonds such as hydrogen bonding, metal coordination, hydrophobicity, van der Waals forces, π-π interactions, halogen bonding, electrostatics, and / or electromagnetic effects. In certain embodiments, the CB is selected from nanoparticles, immunoglobulins, nucleic acids, proteins, oligopeptides, polypeptides, antibodies, fragments of antigenic polypeptides, repibodies, and other homologous materials.

[0609] The compound and conjugate of the present invention may include one or more targeting moiety. That is, the variable cb may have an integer value selected from 1, 2, 3, 4, 5, 1-10, or 1-20.

[0610] In some embodiments, CB is a covalent bond ( for exampleThe peptide comprises two or more independently selected natural or non-natural amino acids conjugated by peptide bonds, and the peptide may comprise 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more natural or non-natural amino acids conjugated by peptide bonds. In some embodiments, the ligand has a shorter amino acid sequence ( for example , fragments of natural proteins or synthetic polypeptide fragments) as well as whole proteins ( for example It includes pre-engineered proteins.

[0611] In some embodiments, CB is an antibody, hormone, drug, or antibody analog that binds to a receptor ( for example , non-IgG), protein, oligopeptide, polypeptide, etc. It is selected from. In certain embodiments, the CB selectively targets a drug in a specific organ, tissue, or cell. In other embodiments, the CB specifically binds to a receptor overexpressed in cancer cells compared to normal cells and may be classified as a monoclonal antibody (mAb) or antibody fragment and a small-molecule non-antibody. Preferably, the CB is selected from peptides identified in a library screen, tumor cell-specific peptides, tumor cell-specific aptamers, tumor cell-specific carbohydrates, tumor cell-specific monoclonal antibodies, polyclonal antibodies, and antibody fragments.

[0612] Exemplary ligands or targeting moietyes include, but are not limited to, carnitine, inositol, lipoic acid, pyridoxal, ascorbic acid, niacin, pantothenic acid, folic acid, riboflavin, thiamine, biotin, vitamin B 12, other water-soluble vitamins (Vitamin B), fat-soluble vitamins (Vitamins A, D, E, K), RGD (Arg-Gly-Asp), NGR (Asn-Gly-Arg), transferrin, VIP (vasoactive enteric peptide) receptor, APRPG (Ala-Pro-Arg-Pro-Gly) peptide, TRX-20 (Thioredoxin-20), integrin, nucleolin, aminopeptidase N (CD13), endoglin, vascular epithelial growth factor receptor, low-density lipoprotein receptor, transferrin receptor, somatostatin receptor, bombesin, neuropeptide Y, luteinizing hormone-releasing hormone receptor, folic acid receptor, epidermal growth factor receptor, transforming growth factor, fibroblast growth factor receptor, asarloglycan protein receptor, galectin-3 receptor, E-selectin receptor, hyaluronic acid receptor, Prostate-specific membrane antigen (PSMA), cholecystokinin A receptor, cholecystokinin B receptor, discoidin domain receptor, mucin receptor, opioid receptor, plasminogen receptor, bradykinin receptor, insulin receptor, insulin-like growth factor receptor, angiotensin AT1 receptor, angiotensin AT2 receptor, granulocyte macrophage colony-stimulating factor receptor (GM-CSF receptor), galactosamine receptor, sigma-2 receptor, delta-like 3 (DLL-3), aminopeptidase P, melanotransferrin, leptin, tetanustoxin Tet1, tetanustoxin G23, RVG (rabies virus glycoprotein) peptide, HER2 (human epidermal growth factor receptor 2), GPNMB (glycoprotein non-transferable b), Ley, CA6, CanAng, SLC44A4 (solute carrier family 44 Member 4), CEACAM5 (Carcinoma embryonic antigen-associated cell adhesion molecule 5), Nectin-4, Carbonic anhydrous enzyme 9, TNNB2, 5T4, CD30, CD37, CD74, CD70, PMEL17, EphA2 (Ephrin A2 receptor), Trop-2, SC-16, Tissue factor, ENPP-3 (AGS-16),SLITRK6 (SLIT and NTRK-like family member 6), CD27, Lewis Y antigen, LIV1, GPR161 (G protein-coupled receptor 161), PBR (peripheral-type benzodiazepine receptor), MERTK (Mer receptor tyrosine kinase) receptor, CD71, LLT1 (lectin-like transcript 1 or CLED2D), interleukin-22 receptor, sigma 1 receptor, peroxisome proliferator-activated receptor, DLL3, C4.4a, cKIT, ephrin A, CTLA4 (cytotoxic T-lymphocyte-associated protein 4), FGFR2b (fibroblast growth factor receptor 2b), N-acetylcholine receptor, gonadotropin-releasing hormone receptor, gastrin-releasing peptide receptor, bone morphogenetic protein receptor-type 1B (BMPR1B), E16 (LAT1, SLC7A5), STEAP1 (of the prostate 6 transmembrane epithelial antigens), 0772P (CA125, MUC16), MPF (MSLN, mesothelin), Napi3b (SLC34A2), Sema5b (semaphorin 5b), ETBR (endothelin type B receptor), MSG783 (RNF124), STEAP2 (6 transmembrane epithelial antigens of prostate 2), TrpM4 (transient receptor translocation cation 5 channel, subfamily M, member 4), CRIPTO (teratoma-derived growth factor), CD21, CD79b, FcRH2 (IFGP4), HER2 (ErbB2), NCA (CEACM6), MDP (DPEP1), IL20R-alpha (IN20Ra), Brevican (BCAN), EphB2R, ASLG659 (B7h), CD276, PSCA (prostate stem cell antigen) Precursor), GEDA, BAFF-R (BR3), CD22 (BL-CAM), CD79a, CXCR5, HLA-DOB, P2X5, CD72, LY64, FcRH1, IRTA2, TENB2, SSTR2, SSTR5, SSTR1, SSTR3, SSTR4, ITGAV (integrin, alpha 5), ​​ITGB6 (integrin, beta 6), MET, MUC1, EGFRvIII,CD33, CD19, IL2RA (Interleukin II Receptor, Alpha), AXL, BCMA, CTA (Cancer Testis Antigen), CD174, CLEC14A, GPR78, CD25, CD32, LGR5 (GPR49), CD133 (Prominin), ASG5, ENPP3 (Ectonucleotide Pyrophosphatase / Phosphodiesterase 3), PRR4 (Proline-Rich Protein 4), GCC (Guanylate Cyclasase 2C), Liv-1 (SLC39A6), CD56, CanAg, TIM-1, RG-1, B7-H4, PTK7, CD138, Claudin, Her3 (ErbB3), RON (MST1R), CD20, TNC (Thenassin C), FAP, DKK-1, CD52, CS1 (SLAMF7), Annexin A1, V-CAM, gp100, MART-1, MAGE-1 (Melanoma Antigen-Encoding Gene-1), MAGE-3 (Melanoma-Associated Antigen 3), BAGE, GAGE-1, MUM-1 (Multiple Myeloma Oncogene 1), CDK4, TRP-1(gp75), TAG-72 (Tumor-Associated Glycoprotein-72), Gangliosides GD2, GD3, GM2, GM3, VEP8, VEP9, My1, VIM-D5, D156-22, OX40, RNAK, PD-L1, TNFR1, TNFR2, etc.

[0613] Target

[0614] In some embodiments, the targets or targets of the molecular recognition element are specifically associated with one or more specific cell or tissue types. In some embodiments, the targets are specifically associated with one or more specific disease states. In some embodiments, the targets are specifically associated with one or more specific developmental stages. For example, cell type-specific markers are typically expressed at a level at least 2 times greater in the cell type than in a reference population of cells. In some embodiments, cell type-specific markers are present at a level at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 50 times, at least 100 times, or at least 1,000 times greater than their average expression in the reference population. Detection or measurement of cell type-specific markers enables distinguishing cell types or types of interest from many, most, or all other types of cells. In some embodiments, the targets may include proteins, carbohydrates, lipids, and / or nucleic acids as described herein.

[0615] In some embodiments, a substance is considered "targeted" when it specifically binds to a targeting moiety, e.g., a nucleic acid targeting moiety. In some embodiments, the targeting moiety, e.g., a nucleic acid targeting moiety, specifically binds to a target under strict conditions.

[0616] In specific embodiments, the conjugates and compounds described herein are one or more targets associated with an organ, tissue, cell, extracellular matrix component, and / or intracellular compartment ( for exampleIt includes a targeting moiety that specifically binds to an antigen. In some embodiments, the conjugates and compounds described herein include a targeting moiety that specifically binds to a target associated with a specific organ or organ system. In some embodiments, the conjugates and compounds described herein include a targeting moiety that specifically binds to one or more intracellular targets (e.g., organelles, intracellular proteins). In some embodiments, the conjugates and compounds described herein include a targeting moiety that specifically binds to a target associated with a diseased organ, tissue, cell, extracellular matrix component, and / or intracellular compartment. In some embodiments, the conjugates and compounds described herein include a specific cell type ( for example , endothelial cell, cancer cell, malignant cell, prostate cancer cell, etc. It includes a targeting moiety that specifically binds to a target associated with ).

[0617] In some embodiments, the conjugates and compounds described herein are one or more specific tissue types ( for example , liver tissue vs. It includes a targeting moiety that binds to a specific target for prostate tissue. In some embodiments, the conjugate and compound described herein are one or more specific cell types ( for example , T cells vs. It includes a targeting moiety that binds to a specific target for B cells. In some embodiments, the conjugate and compound described herein are for one or more specific disease conditions ( for example , tumor cells vs. It includes a targeting moiety that binds to a target specific to healthy cells. In some embodiments, the conjugate and compound described herein comprise one or more specific developmental stages ( for example , stem cells vs. It includes a targeting moiety that binds to a specific target for differentiated cells.

[0618] In some embodiments, the target may be a marker exclusively or primarily associated with one or several cell types, one or several diseases, and / or one or several developmental stages. Cell type-specific markers are typically, for example, a plurality of ( for example (5-10 or greater) expressed at a level at least 2 times greater in a cell type than in a reference population of cells that may consist of a mixture containing cells from different tissues or organs. In some embodiments, the cell type-specific marker is present at a level at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 50 times, at least 100 times, or at least 1000 times greater than its average expression in the reference population. Detection or measurement of the cell type-specific marker enables distinguishing the cell type of interest or types from many, most, or all other types of cells.

[0619] In some embodiments, the target comprises proteins, carbohydrates, lipids, and / or nucleic acids. In some embodiments, the target comprises proteins and / or characteristic portions thereof, e.g., tumor markers, integrins, cell surface receptors, transmembrane proteins, intercellular proteins, ion channels, membrane transport proteins, enzymes, antibodies, chimeric proteins, glycoproteins, etc. In some embodiments, the target comprises carbohydrates and / or characteristic portions thereof, e.g., glycoproteins, sugars ( for example , monosaccharides, disaccharides, polysaccharides), glycocalyx ( in other words , carbohydrate-rich peripheral zones on the outer surface of most eukaryotic cells), etc. ...includes. In some embodiments, the target is a lipid and / or a characteristic part thereof, e.g., oil, fatty acid, glyceride, hormone, steroid ( for example , cholesterol, bile acid), vitamin ( for example, Vitamin E), phospholipids, sphingolipids, lipoproteins, etc. Includes. In some embodiments, the target includes a nucleic acid and / or a characteristic portion thereof, e.g., DNA nucleic acid; RNA nucleic acid; modified DNA nucleic acid; modified RNA nucleic acid; any combination of DNA, RNA, modified DNA, and modified RNA.

[0620] Numerous markers are known in the art. Typical markers include cell surface proteins, e.g., receptors. Exemplary receptors include, but are not limited to, the following: transferrin receptors; LDL receptors; growth factor receptors, such as members of the epidermal growth factor receptor family (e.g., EGFR, Her2, Her3, Her4) or vascular endothelial growth factor receptors, cytokine receptors, cell adhesion molecules, integrins, selectins, and CD molecules. Markers may be molecules present exclusively or in higher amounts on malignant cells, e.g., tumor antigens.

[0621] nanoparticles

[0622] In some embodiments, the targeting moiety is a particle ( for example , target particles), preferably nanoparticles, and optionally targeting nanoparticles attached to targeting molecules capable of specifically or preferably binding to a target. In some embodiments, the targeting particles alone guide the compound of the present invention (e.g., by reinforcement in tumor cells or tissues) and there are no additional targeting molecules attached thereto.

[0623] In this specification, "nanoparticle" means any particle having a diameter of less than 1000 nm. In some embodiments, the therapeutic agent and / or targeting molecule may be associated with the body of the particle, e.g., in a polymer matrix. In some embodiments, the targeting molecule may be covalently associated with the surface of the polymer matrix. In some embodiments, the covalent bond is mediated by a linker. In some embodiments, the therapeutic agent may be associated with the surface of the polymer matrix, encapsulated within it, surrounded by it, and / or dispersed throughout it. For example, refer to US Patent No. 8,246,968 (the entirety of which is incorporated herein).

[0624] Generally, the nanoparticles of the present invention comprise any type of particle. Any particle may be used according to the present invention. In some embodiments, the particles are biodegradable and biocompatible. Generally, the biocompatible substance is not toxic to cells. In some embodiments, the substance is considered biocompatible if its addition to cells results in a cell death below a certain threshold. In some embodiments, the substance is considered biocompatible if its addition to cells does not induce adverse effects. Generally, the biodegradable substance is therapeutically relevant for a period ( for example It is degraded under physiological conditions over a period of time (weeks, months, or years). In some embodiments, the biodegradable substance is a substance that can be degraded by cellular mechanisms. In some embodiments, the biodegradable substance is a substance that can be broken down by chemical processes. In some embodiments, the particle is a substance that is both biocompatible and biodegradable. In some embodiments, the particle is a substance that is biocompatible but not biodegradable. In some embodiments, the particle is a substance that is biodegradable but not biocompatible.

[0625] It is often desirable to use a population of particles that are relatively uniform in terms of size, shape, and / or composition, so that each particle has similar characteristics. For example, at least 80%, at least 90%, or at least 95% of the particles may have a diameter or maximum dimension within 5%, 10%, or 20% of the average diameter or maximum dimension. In some embodiments, the population of particles may be non-uniform with respect to size, shape, and / or composition. Various different particles may be used according to the present invention. In some embodiments, the particles are spherical or ellipsoidal. In some embodiments, the particles are spherical or ellipsoidal. In some embodiments, the particles are flat or plate-shaped. In some embodiments, the particles are cubic or rectangular. In some embodiments, the particles are egg-shaped or elliptical. In some embodiments, the particles are cylindrical, conical, or pyramidal.

[0626] In some embodiments, the particle is an ultraparticle (e.g., a microsphere). Generally, "ultraparticle" refers to any particle having a diameter of less than 1000 μm. In some embodiments, the particle is a picoparticle ( for example , picosphere). Generally, "picoparticle" refers to any particle having a diameter of less than 1 nm. In some embodiments, the particle is a liposome. In some embodiments, the particle is a micelle.

[0627] The particles can be solid or hollow and have one or more layers ( for exampleIt may include nanoshells, nanorings, etc. In some embodiments, each layer has a unique composition and unique properties compared to other layer(s). For example, the particle may have a core / shell structure, wherein the core is one layer and the shell is a second layer. The particle may include a plurality of different layers. In some embodiments, one layer may be substantially cross-linked, and the second layer may not be substantially cross-linked, etc. In some embodiments, one, several, or all different layers may include one or more therapeutic agents or diagnostic agents to be delivered. In some embodiments, one layer includes the agent to be delivered, and the second layer does not include the agent to be delivered, etc. In some embodiments, each individual layer includes a different agent to be delivered or a set of agents.

[0628] In some embodiments, the particles are porous, meaning that the particles contain holes or channels that are typically small relative to the size of the particles. For example, the particles may be porous silica particles, e.g., mesoporous silica nanoparticles, or may have a coating of mesoporous silica (Lin et al., 2005, J. Am. Chem. Soc, 17:4570). The particles have a diameter in the range of about 1 nm to about 50 nm, for example It may have pores with a diameter of about 1 to 20 nm. About 10% to 95% of the volume of the particle may consist of voids within the pores or channels.

[0629] The particles may have a coating layer. The use of a biocompatible coating layer may be advantageous, for example, if the particles contain a substance toxic to cells. Suitable coating materials include, but are not limited to: natural proteins such as bovine serum albumin (BSA); biocompatible hydrophilic polymers such as polyethylene glycol (PEG) or PEG derivatives; phospholipids (PEG); silica; lipids; polymers; carbohydrates such as dextran; other nanoparticles that may be associated with the nanoparticles of the present invention, etc. The coating may be applied or assembled in various ways, for example by immersion, using layer-by-layer techniques, by self-assembly, conjugation, etc. Self-assembly refers to the process of the spontaneous assembly of higher-order structures relying on the natural attractive forces between components of higher-order structures (e.g., molecules) of each other. It typically occurs through the formation of bonds based on size, shape, composition, or chemical properties and the random movement of molecules.

[0630] Examples of polymers include: polyalkylene ( for example , polyethylene), polycarbonate ( for example , poly(l,3-dioxane-2-one)), polyanhydride ( for example , poly(sebaxane anhydride)), polyhydroxy acid ( for example , poly(3-hydroxyalkanoate)), polyfumarate, polycaprolactone, polyamide ( for example , polycaprolactam), polyacetal, polyether, polyester ( for example , polylactide, polyglycolide), poly(orthoester), polyvinyl alcohol, polyurethane, polyphosphazene, polyacrylate, polymethacrylate, polycyanoacrylate, polyurea, polystyrene, and polyamine. In some embodiments, the polymer according to the present invention comprises a polymer approved for human use by the U.S. Food and Drug Administration (FDA) under 21 CFR §177.2600, and includes, without limitation, the following: polyester ( for example, polylactic acid, polyglycolic acid, poly(lactic acid-co-glycolic acid), polycaprolactone, polyvalerolactone, poly(l,3-dioxane-2-one)); polyanhydride ( for example , poly(sebaxane anhydride)); polyether ( for example , polyethylene glycol); polyurethane; polymethacrylate; polyacrylate; and polycyanoacrylate.

[0631] In some embodiments, the particles are non-polymer particles ( for example , metal particles, quantum dots, ceramic particles, polymers containing inorganic materials, bone-derived materials, bone substitutes, virus particles, etc. It may be ). In some embodiments, the therapeutic agent or diagnostic agent to be delivered may be associated with the surface of such non-polymer particles. In some embodiments, the non-polymer particles are aggregates of non-polymer components, such as metal atoms ( for example It is an aggregate of (gold atoms). In some embodiments, the therapeutic agent or diagnostic agent to be delivered may be associated with the surface of an aggregate of a non-polymeric component and / or encapsulated therein, may be surrounded by the above and / or dispersed throughout the above.

[0632] particle ( for exampleNanoparticles, ultrafine particles) can be prepared using any method known in the art. For example, microparticle formulations can be formed by nanoprecipitation, flow focusing fluid channels, spray drying, single and double emulsion solvent evaporation, solvent extraction, phase separation, milling, microemulsion procedures, microfabrication, nanofabrication, sacrificial layers, simple and complex coacervation methods, and other suitable methods. Alternatively or additionally, aqueous and organic solvent synthesis for monodisperse semiconductor, conductive, magnetic, organic, and other nanoparticles has been described (Pellegrino et al., 2005, Small, 1:48; Murray et al., 2000, Ann.Rev. Mat.Sci., 30:545; and Trindade et al., 2001, Chem.Mat., 13:3843).

[0633] Methods for preparing ultrafine particles for the delivery of encapsulated formulations are described in the following literature: literature (e.g., Doubrow, Ed., "Microcapsules and Nanoparticles in Medicine and Pharmacy," CRC Press, Boca Raton, 1992; Mathiowitz et al., 1987, J. Control. Release, 5:13; Mathiowitz et al., 1987, Reactive Polymers, δ See :275; and Mathiowitz et al., 1988, J. Appl. Polymer Sci., 35:755).

[0634] Nucleic acid targeting moiety

[0635] In some embodiments, the targeting moiety includes a nucleic acid targeting moiety.

[0636] Generally, nucleic acid targeting moiety is any polynucleotide that binds to components associated with organs, tissues, cells, extracellular matrix components, and / or intracellular compartments (targets).

[0637] In some embodiments, the nucleic acid targeting moiety is an aptamer. An aptamer is a polynucleotide that binds to a specific target structure associated with a specific organ, tissue, cell, extracellular matrix component, and / or intracellular compartment. Generally, the targeting function of an aptamer is based on the aptamer's three-dimensional structure. In some embodiments, the binding of an aptamer to a target is typically mediated by interactions between the two- and / or three-dimensional structures of both the aptamer and the target. In some embodiments, the binding of an aptamer to a target is not based solely on the aptamer's primary sequence but depends on the three-dimensional structure(s) of the aptamer and / or the target. In some embodiments, the aptamer is a structure that interferes with base pairing ( for example It binds to its target through complementary Watson-Crick base pairing, which is blocked by a hairpin loop.

[0638] In some embodiments, the nucleic acid targeting moiety is a Spiegelmer (PCT Publications WO 98 / 08856, WO 02 / 100442, and WO 06 / 117217). Generally, a Spiegelmer is a synthetic, enantiomer nucleic acid capable of specifically binding to a target ( in other words , enantiomer). Spiegelmers are characterized by structural features that make them insensitive to exo- and endo-nucleases.

[0639] Any nucleic acid targeting moiety capable of specifically binding to a target ( for exampleIt will be recognized that aptamers or spiegelmers may be used according to the present invention. In some embodiments, the nucleic acid targeting moiety to be used according to the present invention may target markers associated with diseases, disorders, and / or pathological conditions. In some embodiments, the nucleic acid targeting moiety to be used according to the present invention may target cancer-associated targets. In some embodiments, the nucleic acid targeting moiety to be used according to the present invention may target tumor markers. Any type of cancer and / or any tumor marker may be targeted using the nucleic acid targeting moiety according to the present invention. For some examples, the nucleic acid targeting moiety may target markers associated with prostate cancer, lung cancer, breast cancer, colorectal cancer, bladder cancer, pancreatic cancer, endometrial cancer, ovarian cancer, bone cancer, esophageal cancer, liver cancer, gastric cancer, brain tumor, cutaneous melanoma, and / or leukemia.

[0640] (nucleic acid targeting moiety and / or functional RNAs to be delivered, described in more detail below, for example The nucleic acids of the present invention (including RNAi-derived entities, ribozymes, tRNAs, etc.) are, without limitation, chemically synthesized, enzymatically synthesized, enzymatically or chemically cleaved from longer precursors, etc. It can be manufactured according to any available technology including.

[0641] Methods for synthesizing RNAs are known in the art ( reference , for example, Gait, MJ (ed.) Oligonucleotide synthesis: a practical approach, Oxford [Oxfordshire], Washington, DC:IRL Press, 1984; and Herdewijn, P. (ed.) Oligonucleotide synthesis: methods and applications, Methods in molecular biology, v. 288 (Clifton, NJ.)Totowa, NJ:Humana Press, 2005).

[0642] Nucleic acids forming a nucleic acid targeting moiety are naturally occurring nucleosides, modified nucleosides, and hydrocarbon linkers inserted between one or more nucleosides ( for example , alkylene) or polyether linker ( for example It may include naturally occurring nucleosides having a PEG linker, modified nucleosides having a hydrocarbon or PEG linker inserted between one or more nucleosides, or a combination thereof. In some embodiments, the nucleotides or modified nucleotides of the nucleic acid targeting moiety may be replaced with hydrocarbon linkers or polyether linkers, and the binding affinity and selectivity of the mononucleotide nucleic acid targeting moiety are not substantially reduced by the substitution ( for example , the dissociation constant of the nucleic acid targeting moiety of the above-mentioned target nucleic acid is approximately 1 x 10⁻⁶ -3 It must not exceed M).

[0643] It will be recognized by those skilled in the art that the nucleic acid according to the present invention may consist entirely of nucleotides of the type found in naturally occurring nucleic acids, or instead may consist of one or more nucleotide analogs, or otherwise have a structure different from that of naturally occurring nucleic acids. U.S. Patent Nos. 6,403,779; 6,399,754; 6,225,460; 6,127,533; 6,031,086; 6,005,087; 5,977,089; and the references therein disclose various specific nucleotide analogs and modifications that may be used. See Crooke, S. (ed.) Antisense Drug Technology: Principles, Strategies, and Applications (1st ed), Marcel Dekker; ISBN: 0824705661; 1st edition (2001) and the references therein. For example, the 2'-modification includes halo, alkoxy, and allyloxy groups. In some embodiments, the 2'-OH group is replaced by a group selected from: H, OR, R, halo, SH, SR, NH2, NHR, NR2, or CN (wherein R is C1-C6 alkyl, alkenyl, or alkynyl, and halo is F, CI, Br, or I). Examples of modified linkers include phosphorothioate and 5'-N-phosphoramidite linkers.

[0644] Nucleic acids comprising various different nucleotide analogs, modified backbones, or non-naturally occurring internucleoside linkers may be used according to the present invention. The nucleic acids of the present invention may comprise natural nucleosides (i.e., adenosine, thymidine, guanosine, cytidine, uridine, deoxyadenosine, deoxythymidine, deoxyguanosine, and deoxycytidine) or modified nucleosides. Examples of modified nucleotides include: base-modified nucleosides ( for example, araccidine, inosine, isoguanosine, nebularine, pseudouridine, 2,6-diaminopurine, 2-aminopurine, 2-thiothymidine, 3-deaza-5-azacitidine, 2'-deoxyuridine, 3-nitropyrrole, 4-methylindole, 4-thiouridine, 4-thiothymidine, 2-aminoadenosine, 2-thiothymidine, 2-thiouridine, 5-bromositidine, 5-ioduridine, inosine, 6-azauridine, 6-chloropurine, 7-deazaadenosine, 7-deazaguanosine, 8-azaudenosine, 8-azidoadenosine, benzimidazole, ML-methyladenosine, pyrrolo-pyrimidine, 2-amino-6-chloropurine, 3-methyladenosine, 5-propynylcytidine, 5-propynyluridine, 5-bromouridine, 5-fluorouridine, 5-methylcytidine, 7-deazadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, 0(6)-methylguanine, and 2-thiocytidine), chemically or biologically modified bases ( for example , methylated bases), modified sugars ( for example , 2'-fluororibose, 2'-aminoribose, 2'-azidoribose, 2'-O-methylribose, L-enantiomer nucleoside arabinose, and hexose), modified phosphate group ( for example , phosphorothioate and 5'-N-phosphoramidite linkers), and combinations thereof. Natural and modified nucleotide monomers for the chemical synthesis of nucleic acids are readily available. In some cases, nucleic acids containing such modifications exhibit improved properties compared to nucleic acids composed solely of naturally occurring nucleotides. In some embodiments, the nucleic acid modifications described herein are nucleases ( for example , exonuclease, endonuclease, etc. It is used to reduce and / or prevent digestion by ). For example, the structure of nucleic acids can be stabilized by including nucleotide analogs at the 3' ends of one or both strands to reduce digestion.

[0645] The modified nucleic acid does not need to be modified uniformly along the full length of the molecule. Different nucleotide modifications and / or backbone structures may exist at various locations in the nucleic acid. Those skilled in the art will acknowledge that nucleotide analogs or other modification(s) may be located at any location(s) of the nucleic acid so that the function of the nucleic acid is substantially unaffected. For one example, the modification may be located at any location of the nucleic acid targeting moiety so that the ability of the nucleic acid targeting moiety to specifically bind to said target is substantially unaffected. The modified region may be at the 5'-end and / or 3'-end of one or both strands. For example, a modified nucleic acid targeting moiety has been used in which approximately 1 to 5 residues at the 5' and / or 3' ends of either strand are nucleotide analogs and / or backbone modifications. The modification may be a 5' or 3' end modification. One or both nucleic acid strands may contain at least 50% unmodified nucleotides, at least 80% unmodified nucleotides, at least 90% unmodified nucleotides, or 100% unmodified nucleotides.

[0646] The nucleic acid according to the present invention may comprise, for example, a modification to a sugar, nucleoside, or internucleoside linker, such as those described below: U.S. Patent Application Publications 2003 / 0175950, 2004 / 0192626, 2004 / 0092470, 2005 / 0020525, and 2005 / 0032733. The present invention encompasses the use of any nucleic acid having any one or more of the modifications described herein. For example, numerous terminal conjugates, for example Lipids, such as cholesterol, lithocholic acid, alumeric acid, or long alkyl branched chains, have been reported to improve cellular uptake. Analogs and modifications, for example, using any suitable assay known in the art, it may be tested to select, for example, that it results in improved delivery of therapeutic agents or diagnostic agents, improved specific binding of the nucleic acid targeting moiety to a target, etc. In some embodiments, the nucleic acid according to the present invention may comprise one or more non-natural nucleoside linkers. In some embodiments, at the 3'-terminus, 5'-terminus, or both 3'- and 5'-terminus of the nucleic acid targeting moiety, one or more internal nucleotides are reversed to link, e.g., a 3'-3' linker or a 5'-5' linker. ' Calculates the connector.

[0647] In some embodiments, the nucleic acid according to the present invention is not synthetic but is a naturally occurring entity isolated from its natural environment.

[0648] Any method can be used to design novel nucleic acid targeting moiety ( for example , U.S. Patent Nos. 6,716,583; 6,465,189; 6,482,594; 6,458,543; 6,458,539; 6,376,190; 6,344,318; 6,242,246; 6,184,364; 6,001,577; 5,958,691; 5,874,218; 5,853,984; 5,843,732; 5,843,653; 5,817,785; 5,789,163; 5,763,177; 5,696,249; 5,660,985; 5,595,877; 5,567,588; and 5,270,163; and see U.S. Patent Application Publications 2005 / 0069910, 2004 / 0072234, 2004 / 0043923, 2003 / 0087301, 2003 / 0054360, and 2002 / 0064780).

[0649] Nucleic acid targeting moiety that binds to proteins, carbohydrates, lipids, and / or nucleic acids can be designed and / or identified. In some embodiments, the nucleic acid targeting moiety can be designed and / or identified for use in the complex of the present invention that binds to proteins and / or characteristic parts thereof, e.g., tumor markers, integrins, cell surface receptors, transmembrane proteins, intercellular proteins, ion channels, membrane transport proteins, enzymes, antibodies, chimeric proteins, etc. In some embodiments, the nucleic acid targeting moiety binds to carbohydrates and / or characteristic parts thereof, e.g., glycoproteins, sugars ( for example It can be designed and / or identified for use in the complex of the present invention that binds to monosaccharides, disaccharides, and polysaccharides), glycocalyx (i.e., carbohydrate-rich peripheral regions on the outer surface of most eukaryotic cells), etc. In some embodiments, the nucleic acid targeting moiety is lipids and / or characteristic parts thereof, e.g. oils, saturated fatty acids, unsaturated fatty acids, glycerides, hormones, steroids ( for example , cholesterol, bile acid), vitamin ( for example It can be designed and / or identified for use in the complex of the present invention that binds to phospholipids, sphingolipids, lipoproteins, etc. (e.g., vitamin E), etc. In some embodiments, the nucleic acid targeting moiety is a nucleic acid and / or a characteristic portion thereof, e.g., DNA nucleic acid; RNA nucleic acid; modified DNA nucleic acid; modified RNA nucleic acid; and DNA, RNA, modified DNA, and modified RNA; etc. Nucleic acids comprising any combination of can be designed and / or identified in the complex of the present invention.

[0650] Nucleic acid targeting moiety (e.g., aptamer or spiegelmer) can be designed and / or identified using any available method. In some embodiments, the nucleic acid targeting moiety is designed and / or identified by identifying the nucleic acid targeting moiety from a mixture of candidate nucleic acids.

[0651] Method for preparing the compound of the present invention

[0652] The compounds and conjugates disclosed in this specification are produced by a simple method of manufacturing ( for example It can be manufactured through (see Examples 1-36). Such a manufacturing method allows for easy purification.

[0653] Accordingly, a method for preparing the compound of the present invention is also provided herein. For example, the compound of the present invention may be prepared as shown in any one of reaction schemes 4, 5, or 6:

[0654] Reaction Equation 4:

[0655]

[0656] Reaction Equation 5:

[0657]

[0658] Reaction Equation 6:

[0659]

[0660] Here, M is Cl or F and;

[0661] X, Y, Ar, R, and n are equal as previously defined.

[0662] intermediate compounds

[0663] In some embodiments, the compound and conjugate disclosed herein have the chemical formula ( IV It can be prepared by a method using an intermediate compound having a structure according to ) or a pharmaceutically acceptable salt thereof:

[0664]

[0665] Here:

[0666] X is hydrogen, O-, -C(R b )(R c )-, or -N(R c )-, preferably -C(R b )(Rc )-and;

[0667] W is -SO2-G and;

[0668] G is a halogen (preferably fluorine), imidazole, or N-methylimidazolium;

[0669] R is a substituent or -L 1' -Z and;

[0670] L 1' is C1-C 200 - It is an alkylene, wherein it optionally comprises at least one of a peptide bond, an amino bond, an ether bond, a triazole bond, a tetrazole bond, a sugar bond, a sulfonamide bond, a phosphonate bond, a sulfo bond, or a dendrimer structure;

[0671] Z is isocyanide, isothiocyanide, 2-pyridyl disulfide, haloacetamide (-NHC(O)CH2-hal), maleimide, diene, alkene, halide, tosylate (TsO - ), aldehyde, sulfonate (R-SO3 - ), Phosphonic acid (-P(=O)(OH)2), ketone, C8-C 10 Cycloalkyl, -OH, -NHOH, -NHNH2, -SH, carboxylic acid (-COOH), acetylene (-C≡CH), azide (-N3), amino (-NH2), sulfonic acid (-SO3H), alkynon derivative (-C(O)C≡CR a but, R a is C1-C 10 A precursor selected from alkyl), and dihydrogen phosphate (-OP(=O)(OH)2);

[0672] n is an integer with a value from 1 to 4;

[0673] Y is N(R C )-dipeptide (e.g., Val-Cit, Val-Ala), -NO2, -OC(O)(CH2) r C(O)R1, -O(CH2) r-Ar1-NO2, -NHOH, -NHNH2, -BR2R3, or -Y'-TG, e.g. -NO2, -OC(O)(CH2) r C(O)R1, -O(CH2) r -Ar1-NO2, -NHNH2, -BR2R3, or -Y'-TG and;

[0674] R 1 is a C1-C6 alkyl;

[0675] r is an integer from 1 to 5;

[0676] Ar 1 is C6-C 20 -Arilen;

[0677] R 2 and R 3 Each is independently hydrogen, C1-C6-alkyl, C1-C6-alkoxy, or hydroxy;

[0678] R a , R b , R c , and R d Each is independently hydrogen or C1-C6 alkyl;

[0679] Y' is -(CH2) x NR"-, -(CH2) x O-, or -(CH2) x S-go;

[0680] R" is hydrogen or C1-C6 alkyl;

[0681] x is an integer of 0 or 1; and

[0682] TG is an inducing agent.

[0683] In some embodiments, the compound and conjugate disclosed herein have the chemical formula ( V )to It can be prepared by a method using an intermediate compound having a different structure or a pharmaceutically acceptable salt thereof:

[0684]

[0685] Here:

[0686] W, L 1' , and Z are equations ( IV Identical to the definition regarding ); and

[0687] TG is an inducing group, such as β-galactoside, β-glucuronide, or a combination of β-galactoside and β-glucuronide.

[0688] In another embodiment, the compound and conjugate disclosed herein are of the formula ( VI )to It can be prepared by a method using an intermediate compound having a different structure or a pharmaceutically acceptable salt thereof:

[0689]

[0690] Here:

[0691] W is the chemical formula ( IV Identical to what is defined for );

[0692] Y is N(R C )-dipeptide (e.g., Val-Cit, Val-Ala), -NO2, -OC(O)(CH2) r C(O)R 1 , -O(CH2) r -Ar 1 -NO2, -NHOH, -NHNH2, -BR 2 R 3 , or -O-TG and;

[0693] R 1 is a C1-C6-alkyl, e.g., NO2, -OC(O)(CH2) r C(O)R 1 , -O(CH2) r -Ar 1 -NO2, -NHOH, -NHNH2, -BR 2 R 3 , or -O-TG and;

[0694] R 1 is a C1-C6-alkyl;

[0695] r is an integer from 1 to 5;

[0696] Ar 1 is phenylene, biphenylene, or naphthalene;

[0697] R 2 and R 3 Each is independently hydrogen, C1-C6-alkyl, C1-C6-alkoxy, or hydroxy;

[0698] R a , R b , R c , and R d Each is independently hydrogen or C1-C6-alkyl; and

[0699] TG is a derivative, β-galactoside, β-glucuronide, or a combination of β-galactoside and β-glucuronide.

[0700] Antibody-drug conjugate (ADC)

[0701] In some embodiments, CB is an antibody, and Q is a drug. Accordingly, the compounds and conjugates disclosed herein can form antibody-drug conjugates (ADCs) by conjugating an antibody to a drug moiety. Antibody-drug conjugates (ADCs) are capable of selectively delivering one or more drug moiety(s) to target tissues, e.g., tumor-associated antigens, due to the ADC's ability to deliver these to diseases, for example , can increase the therapeutic efficacy for treating cancer. Therefore, in certain embodiments, the present invention is for therapeutic use, for example , provides ADCs for the treatment of cancer.

[0702] The ADC of the present invention comprises an antibody connected to one or more drug moieties. The specificity of the ADC is defined by the specificity of the antibody. In one embodiment, the antibody is connected to one or more cytotoxic drug(s) that are delivered internally to cancer cells.

[0703] Examples of drugs that may be used in the ADC of the present invention are provided below. The terms "drug," "formulation," and "drug moiety" are used interchangeably herein. The terms "linked" and "conjugated" are also used interchangeably herein and indicate that the antibody and the moiety are covalently bonded.

[0704] In some embodiments, the ADC has the following chemical formula (chemical formula VII has:

[0705] (DL) n -Ab ( VII )

[0706] Here, Ab is an antibody, and (DL) is a linker-drug moiety. The linker-drug moiety is formed from linker L and drug moiety D. The drug moiety may have, for example, inhibitory cell proliferation, cytotoxicity, or other therapeutic activity against target cells. n is an integer having a value of 1 to about 20, preferably 1 to about 10. Preferably, DL is the formula ( I" It has the structure of ):

[0707]

[0708] ( I" )

[0709] Each Q is an activator independently connected to L' by a heteroatom, preferably O or N;

[0710] Z' is the chemical formula independently in each case ( I" The structure of ) (CB) cb A linker connecting to, a solubilizing group, a reactive group (e.g., a precursor group), a solid surface (e.g., a particle), a stabilizing group, a chelator, a biopolymer (e.g., an immunoglobulin, nucleic acid, protein, oligopeptide, polypeptide, antibody, fragment or repeat of an antigenic polypeptide), an activator, or a detectable moiety, provided that at least one occurrence of Z' is of the chemical formula ( I" The structure of ) (CB)cb Connect to;

[0711] Each L' is a spacer moiety independently attached to SO2 through a heteroatom selected from O, S, and N, preferably O or N, and the cleavage of the bond between L' and SO2 is selected to facilitate the cleavage of the bond between L' and Q to release an activator;

[0712] Each X is independently -O-, -C(R b )(R c )-, or -N(R c )-, preferably -C(R b )(R c )-and;

[0713] E is an integer having a value of 1, 2, or 3, preferably 1;

[0714] Ar is a 6-membered aryl, 6-membered heteroaryl ring;

[0715] Y' is -N(R a )-, -O-, or -S- and;

[0716] At least one X is located in an ortho relation or a para relation with respect to Y' on Ar;

[0717] TG, when cleaved, SO2 and (Q) q -(L') w It is an inducing agent that generates N, O, or S atoms capable of initiating the emission of;

[0718] Each q is an integer having a value of 1 to about 20, preferably 1 to about 10, independently;

[0719] Each of w and x is an integer that independently has a value of 0 or 1;

[0720] Each R a and R c is independently hydrogen or lower alkyl; and

[0721] At least one R b Under the condition that is Z', each Rb is independently Z', hydrogen, or lower alkyl; or

[0722] R b and R c ..., together with the atoms to which they are attached, form a 3-5-membered ring, preferably a 3-4-membered ring;

[0723] However, when w is 0, q is 1.

[0724] In a specific embodiment, each X is located in an ortho relation or a para relation with respect to Y' on Ar.

[0725] In a specific embodiment, at least one X and Y' are located in an ortho relationship with each other on Ar. In a specific such embodiment, E is 2 or 3. In a specific such embodiment where E is 2, two occurrences of X are located in an ortho relationship with respect to Y'.

[0726] In another embodiment, at least one X and Y' are located in a para relationship with each other on Ar. In a specific such embodiment, E is 1.

[0727] In a specific embodiment, at least one X is located in an ortho relation or a para relation with respect to Y'.

[0728] In a specific embodiment, R attached to Ar b Excluding at least one R b represents Z'.

[0729] In some embodiments, n is an integer having a value of 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, or 1. When cb is 1 and n is 1, the drug-to-antibody ratio (DAR) of the ADC is equal to the number of drugs present in (DL). When cb is not 1, the drug-to-antibody ratio (DAR) of the ADC is equal to the ratio of the number of drugs present in (DL) to the number of antibodies present in the conjugate.

[0730] Exemplary drug for conjugation

[0731] The ADC of the present invention provides a targeted therapy that can reduce side effects often seen in anticancer therapy as, for example, one or more activator(s) or drug(s) are delivered to specific cells.

[0732] For example, drugs may be selected from the following groups: erlotinib (Tarceva; Genentech / OSI Pharm.); bortezomib (Velcade; MilleniumPharm.); fulvestrant (Faslodex; AstraZeneca); Sutent (SU11248; Pfizer); letrozole (Femara; Novartis); imatinib mesylate (Gleevec; Novartis); PTK787 / ZK 222584 (Novartis); oxaliplatin (Eloxatin; Sanofi); 5-fluorouracil (5-FU); leucovorin; rapamycin (sirolimus, RAPAMUNE; Wyeth); lapatinib (Tykerb, GSK572016; GlaxoSmithKline); lonafarnib (SCH 66336); Sorafenib (BAY43-9006; Bayer Labs.); Gefitinib (Iressa; Astrazeneca); AG1478, AG1571 (SU 5271; Sugen); Alkylating agents ( for example , thiotepa or CYTOXAN® cyclophosphamide); alkyl sulfonate ( for example , busulfan, improsulban or piposulfan); aziridine ( for example , benzodopa, carboquone, meturedopa or uredopa); ethyleneimine, methylmelamine, altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, trimethylolmelamine; acetogenin ( for example , bulatacin or bulatacinone); camptothecin containing the synthetic analog topotecan; bryostatin; calistatin; CC-1065 (including adozelesin, carzelesin or its synthetic analogues of viselesin); cryptophycin ( for example, cryptophysin 1 or cryptophysin 8); dolastatin; duocarmycin (including synthetic analogs, KW-2189, and CB1-TM1); eluterobin; pancratistatin; sarcodictine; spongestatin; nitrogen mustard ( for example , chlorambucil, chlornafazine, colophamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novelbicin, phenesterin, prednimustine, trophosphamide or uracil mustard); nitrosourea ( for example , camustine, chlorosotocin, potemustine, lomustine, nimustine, or ranimnustine); antibiotics ( for example , dynemycin including dynemycin A as a calichiamycin or endiin antibiotic selected from calichiamycin gamma 1 I and calichiamycin omega 1 I); bisphosphonate ( for example , clodronate); esperamycin, neocarzinostatin chromophore or related pigment protein endiine antibiotic chromophore, aclaxinomycin, actinomycin, antramycin, azacerin, bleomycin, cactinomycin, carrabicin, carninomycin, carzinophylline, cromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, ADRLIMYCIN® doxorubicin ( for example , morpholino-doxorubicin, cyanomopolino-doxorubicin, 2-pyrrolino-doxorubicin, liposomal doxorubicin or deoxydoxorubicin), epirubicin, esorubicin, marcelomycin, mitomycin ( for example , mitomycin C, mycophenolic acid, nogalamycin, olibomycin, peflomycin, portpyromycin, puromycin, quellamycin, rhodorubicin, streptonigreen, streptozosin, tubercidin, uvenimex, genostatin or rhodorubicin); anti-metabolites ( for example , 5-fluorouracil (5-FU)); folic acid analog ( for example, denopterin, methotrexate, pteropterin, or trimetrexate); purine analogues ( for example , fludarabine, 6-mercaptopurine, thiamiphrine, or thiaguanine); pyrimidine analogs ( for example , ancitabine, azacitidine, 6-azauridine, camofur, cytarabine, dideoxyuridine, doxyfluridine, enositabine or fluoxuridine); androgen ( for example , callosterone, dromostanolone propionate, epithiostanol, mepitiostan, or testolactone); anti-adrenal ( for example , aminoglutechimide, mitothan, or trilostan); folic acid supplements ( for example , folic acid); aceglatone; aldophosphamide glycoside; aminolevelic acid; eniluracil; amsacrin; vestrabusil; bisantren; edatlaxate; depopamine; demecolsin; diaziquone; L-fornitin; elliptinium acetate; epotillone; etogluside; gallium nitrate; hydroxyurea; lentinan; ronidinine; meitansinoid ( for example , meitansin or ansamitosin; trichothecen ( for example , T-2 toxin, veraculin A, loridin A or anguidin); mitoguazone; mitoxantrone; mophidanmol; nitraerin; pentostatin; phenamet; pirarubicin; rosoxantrone; 2-ethylhydrazide; procarbazine; PSK® polysaccharides; rasozoxic acid; lyzoxin; sizopyran; spirogermanium; tenuaginic acid; triaziquone; 2,2',2"-trichlorotriethylamine; trichothecene (especially, T-2 toxin, veraculin A, loridin A or anguidin); urethane; vindecin; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; acitosine; arabinoside ('Ara-C'); cyclophosphamide; thiotepa; taxoid ( for example , Taxol® Paclitaxel (Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANE TMCremopore-free, albumin-engineered nanoparticle formulation of paclitaxel, American Pharmaceutical Partners, Schaumber, I11. or TAXOTERE® docetaxel ((Rhone-Poulenc Rorer, Antony, France))); chloranbucil; gemcitabine; 6-thioguanine; mercaptopurine; platinum analogues ( for example , cisplatin or carboplatin); vinblastine; platinum; etoposide, ifosfamide; mitoxantrone; vincristine; NAVELBINE® vinorelbine; Novantron; tenifoside; dedatrexate; daunomycin; aminopterin; Xeloda; ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DFMO); retinoid ( for example , retinoic acid); capecitabine; and its pharmaceutically acceptable salt, its solvate, or its derivative.

[0733] Mitosis inhibitor

[0734] In some embodiments, the linker of the present invention may form an ADC for the treatment of cancer by conjugating an antibody to one or more mitotic inhibitor(s). The term “mitotic inhibitor” as used herein refers to a cytotoxic and / or therapeutic agent that blocks mitosis or cell division, or biological processes particularly important to cancer cells. Mitotic inhibitors disrupt microtubules so that cell division is often prevented by affecting microtubule polymerization or microtubule depolymerization. Accordingly, in certain embodiments, the antibody is conjugated to one or more mitotic inhibitor(s) that disrupt microtubule formation by inhibiting tubulin polymerization. In one embodiment, the mitotic inhibitor used in the ADC of the present invention is Taxol ® (Paclitaxel), Taxotere ® (docetaxel), or Ixempra ®(Ixabefilone). Examples of mitotic inhibitors that may be used in the ADCs disclosed herein are provided below. The genus of mitotic inhibitors includes the aforementioned auristatin.

[0735] Auristatin ( Auristatins )

[0736] The linker of the present invention may be used to conjugate an antibody to at least one auristatin. Auristatin represents a group of dolastatin analogs generally known to inhibit cell division by possessing anticancer activity by interfering with microtubule kinetics and GTP hydrolysis. For example, auristatin E (US Patent No. 5,635,483) is a synthetic analog of the marine natural product dolastatin 10, which is a compound that inhibits tubulin polymerization by binding to the same site on tubulin as the anticancer drug vincristine (GRPettit, Prog.Chem.Org.Nat. Prod, 70:1-79 (1997)). Dolastatin 10, auristatin PE, and auristatin E are linear peptides having four amino acids, three of which are unique to the class of dolastatin compounds. Exemplary embodiments of an auristatin subclass of mitotic inhibitors are, without limitation, the following: secondIncludes: monomethylauristatin D (MMAD or auristatin D derivative), monomethylauristatin E (MMAE or auristatin E derivative), monomethylauristatin F (MMAF or auristatin F derivative), auristatin F phenylenediamine (AFP), auristatin EB (AEB), auristatin EFP (AEFP), and 5-benzoylvaretic acid-AE ester (AEVB). The synthesis and structure of the auristatin derivatives are described below: US Patent Application Publications Nos. 2003-0083263, 2005-0238649 and 2005-0009751; International Patent Publication No. WO 04 / 010957, International Patent Publication No. WO 02 / 088172, and US Registered Patent Nos. 6,323,315; 6,239,104; 6,034,065; 5,780,588; 5,665,860; 5,663,149; 5,635,483; 5,599,902; 5,554,725; 5,530,097; 5,521,284; 5,504,191; 5,410,024; 5,138,036; 5,076,973; 4,986,988; 4,978,744; 4,879,278; 4,816,444; and 4,486,414 (each of which is incorporated herein by reference).

[0737] Dolastatin ( Dolastatins )

[0738] The linker of the present invention can form an ADC by conjugating an antibody to at least one dolastatin. Dolastatin is a short peptide compound isolated from the Indian Ocean sea rabbit *Dolabella auricularia* (Pettit etc. , J. Am. Chem.Soc.(See , 1976, 98, 4677). Examples of dolastatin include dolastatin 10 and dolastatin 15. Dolastatin 15, a 7-subunit depsipeptide derived from *Dolabella origularia*, is a 5-subunit peptide obtained from the same organism and is a potent anti-cell division agent structurally related to dolastatin 10, an anti-tubulin agent. Thus, in one embodiment, the ADC of the present invention comprises an antibody, a linker as described herein, and at least one dolastatin. The auristatin described above is a synthetic derivative of dolastatin 10.

[0739] Meitansinoid ( Meytansinoids )

[0740] The linker of the present invention can form an ADC by conjugating an antibody to at least one metansinoid. Metansinoids are potent antitumor agents originally isolated from members of the higher plant families Celastraceae, Rhamnaceae, and Euphorbiaceae, as well as some species of mosses (Kupchan et al., J. Am. Chem.Soc.94:1354-1356

[1972] ; Wani et al., J. Chem.Soc.Chem.Commun 390:

[1973] ; Powell et al., J. Nat. Prod.46:660-666

[1983] ; Sakai et al., J. Nat. Prod.51:845-850

[1988] ; and Suwanborirux et al., Experientia 46:117-120

[1990] ). The evidence suggests that meitansinoids inhibit mitosis by inhibiting the polymerization of the microtubule protein tubulin, thereby preventing the formation of microtubules ( for example , US Patent No. 6,441,163 and Remillard etc. , Science(See , 189, 1002-1005 (1975)). Metansinoids have been shown to inhibit tumor cell growth in vitro using cell culture models and in vivo using laboratory animal systems. Furthermore, the cytotoxicity of metansinoids is 1,000-fold greater than that of conventional chemotherapy agents, e.g., methotrexate, daunorubicin, and vincristine ( for example , refer to US Patent No. 5,208,020).

[0741] Meitansinoids include the following: meitansin, meitansinol, C-3 esters of meitansinol, and other meitansinol analogs and derivatives ( for example , see US Patent Nos. 5,208,020 and 6,441,163, each incorporated herein by reference). C-3 esters of metancinol may be of natural or synthetic origin. Furthermore, both naturally occurring and synthetic C-3 metancinol esters may be classified as C-3 esters having a simple carboxylic acid or as C-3 esters having a derivative of N-methyl-L-alanine, the latter being more cytotoxic than the former. Synthetic metancinoid analogs are, for example, described below: Kupchan etc. , J. Med. Chem. , 21, 31-37 (1978).

[0742] Suitable metansinoids used in the ADC of the present invention may be isolated from natural sources, produced synthetically, or produced semi-synthetically. Furthermore, metansinoids may be modified in any suitable manner as long as sufficient cytotoxicity is preserved in the ultimate conjugate molecule. The structure of an exemplary metansinoid, metansin (DM1), is provided below.

[0743]

[0744] Mertansin (DM1)

[0745] Representative examples of metansinoids include, but are not limited to, the following: DM1 (N2'-deacetyl-N2'-(3-mercapto-1-oxopropyl)-metansine; also known as metansine, drug metansinoid 1; ImmunoGen, Inc.; Chari etc. (1992) Cancer Res See 52:127), DM2, DM3 (N2'-deacetyl-N2'-(4-mercapto-1-oxopentyl)-metansin), DM4 (4-methyl-4-mercapto-1-oxopentyl)-metansin), and metansinol (synthetic metansinoid analogs). Other examples of metansinoids are described in US Patent No. 8,142,784, which is incorporated herein by addition.

[0746] Ansamitocins are a group of metancinoid antibiotics isolated from various bacterial sources. These compounds possess potent antitumor activity. Representative examples include, but are not limited to, ansamitocin P1, ansamitocin P2, ansamitocin P3, and ansamitocin P4.

[0747] Plant alkaloids

[0748] The linker of the present invention comprises at least one plant alkaloid, an antibody, for example , can be used to conjugate to taxanes or vinca alkaloids. Plant alkaloids are chemotherapy agents made from specific types of plants. Vinca alkaloids are made from the periwinkle plant *Catharanthus rosus*, while taxanes are made from the bark of the Pacific yew tree *Taxus*. Both vinca alkaloids and taxanes are also known as antimicrotubule agents and are described in more detail below.

[0749] Taksan ( taxanes )

[0750] The linker of the present invention may be used to conjugate an antibody to at least one taxane. The term "taxane" as used herein refers to a class of antineoplastic agents having a mechanism of microtubule action and a structure comprising a taxane ring structure and stereospecific side chains necessary for cell proliferation inhibitory activity. Within the term "taxane," various known derivatives are also included, including both hydrophilic derivatives and hydrophobic derivatives. Taxane derivatives include, but are not limited to: galactose and mannose derivatives described in International Patent Application No. WO 99 / 18113; piperazino and other derivatives described in WO 99 / 14209; taxane derivatives described in WO 99 / 09021, WO 98 / 22451, and US Registered Patent No. 5,869,680; 6-thio derivatives described in WO 98 / 28288; sulfenamide derivatives described in US Registered Patent No. 5,821,263; and taxol derivatives described in US Patent No. 5,415,869 (each of which is incorporated herein by reference). Taxane compounds have also been previously described in the following U.S. registered patents: U.S. Patent Nos. 5,641,803, 5,665,671, 5,380,751, 5,728,687, 5,415,869, 5,407,683, 5,399,363, 5,424,073, 5,157,049, 5,773,464, 5,821,263, 5,840,929, 4,814,470, 5,438,072, 5,403,858, 4,960,790, 5,433,364, 4,942,184, 5,362,831, 5,705,503, and 5,278,324 (all of these are explicitly incorporated by reference). Additional examples of taxanes include, but are not limited to, the following: docetaxel (Taxotere ® ; Sanofi Aventis), paclitaxel (Abraxane ® or Taxol ®; Abraxis Oncology), and nanoparticle paclitaxel (ABI-007 / Abraxene ® ; Abraxis Bioscience).

[0751] In one embodiment, the linker of the present invention may be used to conjugate an antibody to at least one docetaxel. In one embodiment, the linker of the present invention may be used to conjugate an antibody to at least one paclitaxel.

[0752] Vinca alkaloids ( Vinca Alkaloids)

[0753] In one embodiment, the linker of the present invention may be used to conjugate an antibody to at least one vinca alkaloid. Vinca alkaloids are a class of cell-cycle-specific drugs that act by inhibiting the ability of cancer cells to divide by acting on tubulin and preventing the formation of microtubules. Examples of vinca alkaloids that may be used in the ADC of the present invention include, but are not limited to, the following: vindecin sulfate, vincristine, vinblastine, and vinorelbine.

[0754] antitumor antibiotics

[0755] The linker of the present invention may be used to conjugate an antibody to one or more antitumor antibiotic(s) for the treatment of cancer. As used herein, the term “antitumor antibiotic” refers to an antineoplastic drug made of microorganisms that blocks cell growth by interfering with DNA. Often, antitumor antibiotics degrade DNA strands or slow down or stop DNA synthesis. Examples of antitumor antibiotics that may be included in the ADC disclosed herein include, but are not limited to: actinomycin ( for example , pyrrolo[2,1-c][1,4]benzodiazepine), anthracyclines, calikiamycin, and duocarmycin, these are described in more detail below.

[0756] Actinomycin ( Actinomycines )

[0757] The linker of the present invention may be used to conjugate an antibody to at least one actinomycin. Actinomycin is a subclass of antitumor antibiotics isolated from bacteria of the genus Streptomyces. Representative examples of actinomycin include, but are not limited to: actinomycin D (Cosmegen [also known as actinomycin, dactinomycin, actinomycin IV, and actinomycin C1], Lundbeck, Inc.), antramycin, kicamycin A, DC-81, mazetramycin, neotramycin A, neotramycin B, prototramycin, protramycin B, SG2285, sibanomycin, sibiromycin, and tomycin. In one embodiment, D is a pyrrolobenzodiazepine (PBD). Examples of PBD include, but are not limited to: antramycin, kicamycin A, DC-81, mazetramycin, neotramycin A, neotramycin B, prototramycin, protramycin B, SG2000 (SJG-136), SG2202 (ZC-207), SG2285 (ZC-423), sibanomycin, sibiromycin, and tomycin. Thus, in one embodiment, D is actinomycin, for example , actinomycin D, or PBD, for example , is a pyrrolobenzodiazepine (PBD) dimer.

[0758] The structure of a PBD can be found, for example, in the following: US Patent Application Publications Nos. 2013 / 0028917 and 2013 / 0028919, and WO 2011 / 130598 A1 (each of which is incorporated herein by reference in its entirety). The general structure of a PBD is provided below.

[0759]

[0760] PBD differs in the number, type, and position of substituents in both its aromatic A ring and pyrrolo C ring, as well as in the saturation of the C ring. In the B-ring, at the N10-C11 positions, which are the electrophilic centers responsible for alkylating DNA, there are typically imines (N=C), carbinolamines (NH-CH(OH)), or carbinolamine methyl ethers (NH-CH(OMe)). All known natural products have an (S)-position at the chiral C11α position, providing a right-handed twist when viewed from the C ring toward the A ring. Further examples of PBDs that can be conjugated to antibodies via the linkers disclosed herein may be found, for example, in the following: US Patent Application Publications Nos. 2013 / 0028917 A1 and 2013 / 0028919 A1, US Registered Patent No. 7,741,319 B2, and WO 2011 / 130598 A1 and WO 2006 / 111759 A1 (each of which is incorporated herein by reference in its entirety).

[0761] Anthracycline ( Anthracyclines )

[0762] The linker of the present invention may be used to conjugate an antibody to at least one anthracycline. Anthracyclines are a subclass of antitumor antibiotics isolated from bacteria of the genus Streptomyces. Representative examples include, but are not limited to: daunorubicin (Ceruvidin, Bedford Laboratories), doxorubicin (Adriamycin, Bedford Laboratories; also known as doxorubicin hydrochloride, hydroxydaunorubicin, and Rubex), epirubicin (Elense, Pfizer), and idarubicin (Idamycin; Pfizer Inc.). Thus, in one embodiment, D is an anthracycline, for example , it is doxorubicin.

[0763] Caliquiamycin( Calicheamicins )

[0764] The linker of the present invention can be used to conjugate an antibody to at least one calichiamycin. Calichiamycin is a class of endigenic antibiotics derived from the soil organism Micromonospora echinospora. By binding to the narrow groove of DNA and inducing double-stranded DNA disruption, calichiamycin increases 100-fold compared to other chemotherapeutic agents having Cell death is obtained (Damle et al. (2003) Curr Opin Pharmacol 3:386). The preparation of calikiamycin that can be used as a drug conjugate in the present invention is described below: See US Patent Nos. 5,712,374; 5,714,586; 5,739,116; 5,767,285; 5,770,701; 5,770,710; 5,773,001; and 5,877,296. Structural analogs of calichiamycin that may be used include, but are not limited to: γ1 I, α2 I, α3 I, N-acetyl-γ1 I, PSAG and θI I (Hinman et al., Cancer Research 53:3336-3342 (1993), Lode et al., Cancer Research 58:2925-2928 (1998) and the aforementioned US Patent Nos. 5,712,374; 5,714,586; 5,739,116; 5,767,285; 5,770,701; 5,770,710; 5,773,001; and 5,877,296). Thus, in one embodiment, D is calichiamycin.

[0765] Duocarmycin ( Duocarmycins )

[0766] The linker of the present invention can be used to conjugate an antibody to at least one Duocarmycin. Duocarmycin Streptomyces Streptomyces )insideIt is a subclass of antitumor antibiotics isolated from bacteria. (See Nagamura and Saito (1998) Chemistry of Heterocyclic Compounds, Vol. 34, No. 12). Duocarmycin binds to the narrow groove of DNA and alkylates the nucleobase adenine at the N3 position (Boger (1993) Pure and Appl Chem 65(6):1123; and Boger and Johnson (1995) PNAS USA 92:3642). Synthetic analogs of duocarmycin include, but are not limited to: adozelesin, beizelesin, and carzelesin. Thus, in one embodiment, D is duocarmycin.

[0767] Other antitumor antibiotics

[0768] In addition to those mentioned above, additional antitumor antibiotics that may be used in the ADC of the present invention include bleomycin (blenoxan, Bristol-Myers Squibb), mitomycin, and pilicamycin (also known as mitramycin).

[0769] Immunomodulators

[0770] In some embodiments, the linker of the present invention may be used to conjugate an antibody to at least one immunomodulator. As used herein, the term “immunomodulator” refers to a agent capable of stimulating or modifying an immune response. In one embodiment, the immunomodulator is an immunomodulator that enhances the immune response of a subject. In some embodiments, the immunomodulator is an immunosuppressant that prevents or reduces the immune response of a subject. The immunomodulator may modulate bone marrow cells (monocytes, macrophages, dendritic cells, megakaryocytes, and granulocytes) or lymphoid cells (T cells, B cells, and natural killer (NK) cells) and any additional differentiated cells thereof. Representative examples include, but are not limited to: Bacillus calmet-guerin (BCG) and levamisole (ergamisole). Other examples of immunomodulators that may be used in the ADC of the present invention include, but are not limited to: cancer vaccines, cytokines, and immunomodulatory gene therapies.

[0771] cancer vaccine

[0772] The linker of the present invention may be used to conjugate an antibody to a cancer vaccine. As used herein, the term "cancer vaccine" refers to a composition (e.g., tumor antigen and cytokine) that induces a tumor-specific immune response. The response is induced from the subject's own immune system by administering the cancer vaccine, or, in the case of the present invention, by administering an ADC comprising an antibody and a cancer vaccine. In a preferred embodiment, the immune response results in the eradication of tumor cells in the body (e.g., primary or metastatic tumor cells). The use of the cancer vaccine generally involves the administration of a specific antigen or group of antigens present on the surface of specific cancer cells, for example, or on the surface of a specific infectious agent shown to promote cancer formation. In some embodiments, the use of the cancer vaccine is for prophylactic purposes, while in other embodiments, the use is for therapeutic purposes. Non-limiting examples of cancer vaccines that may be used in the ADCs disclosed herein include recombinant bivalent human papillomavirus (HPV) vaccine types 16 and 18 vaccine (Cervarix, GlaxoSmithKline), recombinant quadrivalent human papillomavirus (HPV) types 6, 11, 16, and 18 vaccine (Gardasil, Merck & Company), and ciflucel-T (Provenge, Dendreon). Thus, in one embodiment, D is a cancer vaccine that is an immunostimulant or an immunosuppressant.

[0773] Cytokines ( Cytokines )

[0774] The linker of the present invention can be used to conjugate an antibody to at least one cytokine. The term "cytokine" generally refers to a protein released by a cell population that acts on another cell as an intercellular mediator. Cytokines directly stimulate immune effector cells and stromal cells at the tumor site and enhance tumor cell recognition by cytotoxic effector cells (Lee and Margolin (2011) Cancers 3:3856). Numerous studies using animal tumor models have demonstrated that cytokines possess broad antitumor activity, which has translated into numerous cytokine-based approaches for cancer therapy (Lee and Margolin (ibid.)). Numerous cytokines, including GM-CSF, IL-7, IL-12, IL-15, IL-18, and IL-21, have recently entered clinical trials for patients with advanced cancer (Lee and Margolin (ibid.)).

[0775] Examples of cytokines that may be used in the ADC of the present invention include, but are not limited to: parathyroid hormone; thyroxine; insulin; proinsulin; relaxin; prorelaxin; glycoprotein hormones such as follicle-stimulating hormone (FSH), thyroid-stimulating hormone (TSH), and luteinizing hormone (LH); hepatic growth factor; fibroblast growth factor; prolactin; placental lactogen; tumor necrosis factor; Müllerian-inhibitory substance; mouse gonadotropin-associated peptides; inhibin; activin; vascular endothelial growth factor; integrin; thrombopoietin (TPO); nerve growth factor such as NGF; platelet-growth factor; transforming growth factor (TGFs); insulin-like growth factor-I and-II; erythropoietin (EPO); osteoinducible factor; interferons such as interferon α, β, and γ, colony-stimulating factor (CSF); Granulocyte-macrophage-C-SF (GM-CSF); and granulocyte-CSF (G-CSF); interleukins (ILs) such as IL-1, IL-LA, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-11, IL-12; tumor necrosis factor; and other polypeptide factors including LIF and Kit ligand (KL). As used herein, the term cytokine includes proteins from natural sources or from recombinant cell cultures and biologically active equivalents of natural sequence cytokines. Thus, in one embodiment, D is a cytokine.

[0776] Colony-stimulating factor (CSF)

[0777] The linker of the present invention can be used to conjugate an antibody to at least one colony-stimulating factor (CSF). Colony-stimulating factor (CSF) is a growth factor that assists the bone marrow in the production of red blood cells. Some cancer treatments ( for exampleBecause chemotherapy can affect leukocytes (which help fight infection), colony-stimulating factors may be introduced to support leukocyte levels and help strengthen the immune system. Colony-stimulating factors may also be used after a bone marrow transplant to aid in the generation of novel bone marrow-initiated leukocytes. Representative examples of CSFs that may be used in the ADCs disclosed herein include, but are not limited to, erythropoietin (epoetin), filgrastim (Neopogen, also known as granulocyte colony-stimulating factor (G-CSF); Amgen, Inc.), sargramostim (leukocyte-macrophage colony-stimulating factor and GM-CSF; Genzyme Corporation), promegapoietin, and ofrelbekin (recombinant IL-11; Pfizer, Inc.). Thus, in one embodiment, D is a CSF.

[0778] Gene therapy

[0779] The linker of the present invention can be used to conjugate an antibody to at least one nucleic acid (directly or indirectly for a carrier) for genomic therapy. Gene therapy generally refers to the introduction of genetic material into cells, whereby the genetic material is designed to treat a disease. Because it belongs to immunomodulatory agents, gene therapy is used to stimulate the natural ability of a target to inhibit cancer cell proliferation or to kill cancer cells. In one embodiment, the ADC of the present invention is a cancer-associated mutated or other dysfunctional ( for exampleIt includes a nucleic acid-encoded functional or therapeutic gene used to replace a (truncated) gene. In other embodiments, the ADC of the present invention includes a nucleic acid that encodes a therapeutic protein or provides for the production thereof to treat cancer. The nucleic acid encoding the therapeutic gene may be conjugated directly to an antibody, or alternatively, may be conjugated to an antibody through a carrier. Examples of carriers that may be used to deliver nucleic acids for gene therapy include, but are not limited to, the following: viral vectors or liposomes.

[0780] alkylating agent

[0781] The linker of the present invention may be used to conjugate an antibody to one or more alkylating agents. An alkylating agent is a class of antineoplastic compounds that attach an alkyl group to DNA. Examples of alkylating agents that may be used in the ADC of the present invention include, but are not limited to, the following: alkyl sulfonates, ethyleneimines, methylamine derivatives, epoxides, nitrogen mustard, nitrosoureas, triazines, and hydrazines.

[0782] alkyl sulfonate

[0783] The linker of the present invention can be used to conjugate an antibody to at least one alkyl sulfonate. The alkyl sulfonate has the general formula: R―SO2―O―R 1 It is a subclass of alkylating agents having: where R and R 1 is typically an alkyl or aryl group. A representative example of an alkyl sulfonate is busulpan (Myleran ® , GlaxoSmithKline; Busulfex IV ® , PDL BioPharma, Inc.).

[0784] Nitrogen mustard

[0785] The linker of the present invention can be used to conjugate an antibody to at least one nitrogen mustard. Representative examples of subflectors of anticancer compounds include, but are not limited to, the following: chlorambucil (Leukeran ® , GlaxoSmithKline), cyclophosphamide (Cytoxan ® , Bristol-Myers Squibb; Neosarr, Pfizer, Inc.), estramustine (estramustine phosphate sodium or Estracyt ® ), Pfizer, Inc.), Ifosfamide (Ifex ® , Bristol-Myers Squibb), mechlorethamine (Mustargen ® , Lundbeck Inc.), and Melphalan (Alkeran ® or L-Pam ® or phenylalanine mustard; GlaxoSmithKline).

[0786] Nitrosourea

[0787] The linker of the present invention may be used to conjugate an antibody to at least one nitrosourea. Nitrosourea is a subclass of lipid-soluble alkylating agents. Representative examples include, but are not limited to, the following: Camustine (BCNU [BiCNU, N , N - Bis (2-chloroethyl)- N -Nitrosourea, or 1,3- bis (also known as (2-chloroethyl)-1-nitrosourea], Bristol-Myers Squibb), potemustine (Muphoran ® Also known as), lomustine (CCNU or 1-(2-chloro-ethyl)-3-cyclohexyl-1-nitrosourea, Bristol-Myers Squibb), nimustine (also known as ACNU), and streptozosin (Zanosar ® , Teva Pharmaceuticals).

[0788] Triazine and Hydrazine

[0789] The linker of the present invention may be used to conjugate an antibody to at least one triazine or hydrazine. Triazines and hydrazines are subclasses of nitrogen-containing alkylating agents. In some embodiments, these compounds may spontaneously degrade or be metabolized to promote the transfer of alkyl groups to nucleic acids, peptides, and / or polypeptides, thereby producing alkyl diazonium intermediates that cause mutagenic, carcinogenic, or cytotoxic effects. Representative examples include, but are not limited to: dacarbazine (DTIC-Dome, Bayer Healthcare Pharmaceuticals Inc.), procarbazine (Mutalane ® , Sigma-Tau Pharmaceuticals, Inc.), and temozolomide (Temodar ® , Schering Plough).

[0790] Other alkylating agents

[0791] The linker of the present invention may be used to conjugate an antibody to at least one ethyleneimine, methylamine derivative, or epoxide. Ethyleneimine is a subclass of alkylating agents that typically contains at least one aziridine ring. Epoxide represents a subclass of alkylating agents characterized by a cyclic ether having only three ring atoms.

[0792] Representative examples of ethyleneimines include, but are not limited to, thiofeta (Tioplex, Amgen), diaziquone (also known as aziridinyl benzoquinone (AZQ)), and mitomycin C. Mitomycin C is a natural product containing an aziridine ring that appears to induce cytotoxicity through cross-linked DNA (Dorr RT, et al., Cancer Res. 1985; 45:3510; Kennedy KA, et al., Cancer Res. 1985; 45:3541). Representative examples of methylamine derivatives and their analogs include, but are not limited to, hexamethylamine and altretamine (Hexalen, MGI Pharma, Inc.), also known as hexastat. Representative examples of epoxides of this class of anticancer compounds include, but are not limited to, dianhydrogalactitol. Dianhydrogalactitol (1,2:5,6-dianhydrodulcitol) is chemically related to aziridine and generally promotes the transfer of alkyl groups through a similar mechanism as described above. Dibromodulcitol is hydrolyzed to dianhydrogalactitol and is therefore a prodrug for epoxides (Sellei C, et al. Cancer Chemother Rep. 1969; 53:377).

[0793] Anti-angiogenic agents

[0794] In some embodiments, the linker of the present invention may be used to conjugate an antibody to at least one anti-angiogenic agent. Anti-angiogenic agents inhibit the growth of new blood vessels. Anti-angiogenic agents exert their effects in various ways. In some embodiments, these agents interfere with the ability of growth factors to reach their targets. For example, vascular endothelial growth factor (VEGF) is one of the primary proteins involved in initiating angiogenesis by binding to specific receptors on the cell surface. Therefore, certain anti-angiogenic agents that prevent the interaction between VEGF and its cognate receptors prevent VEGF from initiating angiogenesis. In other embodiments, these agents interfere with intracellular signaling cascades. For example, if a specific receptor on the cell surface is triggered, a cascade of other chemical signals is initiated to promote blood vessel growth. Thus, certain enzymes, for example, some tyrosine kinases, are known to promote intracellular signaling cascades that contribute to cell proliferation and are targets for cancer treatment. In other embodiments, these agents interfere with the intercellular signaling cascade. Also, in other embodiments, these agents directly interfere with the growth of blood vessel cells, thereby inactivating specific targets that activate or promote cell growth. Inhibitory properties of angiogenesis were found in over 300 substances, along with numerous direct and indirect inhibitory effects.

[0795] Representative examples of anti-angiogenic agents that may be used in the ADC of the present invention include, but are not limited to: angiostatin, ABX EGF, C1-1033, PKI-166, EGF vaccine, EKB-569, GW2016, ICR-62, EMD 55900, CP358, PD153035, AG1478, IMC-C225 (Erbitux), ZD1839 (Iressa), OSI-774, erlotinib (Tarceva), angiostatin, Arestin, endostatin, BAY 12-9566 and w / fluorouracil or doxorubicin, canstatin, carboxyamidotriozole and paclitaxel, EMD121974, S-24, vitaxin, dimethylxanthene acetate, IM862, interleukin-12, Interleukin-2, NM-3, HuMV833, PTK787, RhuMab, Angiozyme (Ribozyme), IMC-1C11, Neovastat, Marimstat, Prinomastat, BMS-275291, COL-3, MM1270, SU101, SU6668, SU11248, SU5416, Paclitaxel, Gemcitabine and Cisplatin, and Irinotecan and Cisplatin and Radiation, Tecogalan, Temozolomide and PEG Interferon α2b, Tetrathiomolybdate, TNP-470, Thalidomide, CC-5013 and Taxotere, Tumstatin, 2-Methoxyestradiol, VEGF Trap, mTOR Inhibitors (Dephorolimus, Everolimus (Afinitor, Novartis Pharmaceutical Corporation), and Temsirolimus (Torisel, Pfizer, Inc.), tyrosine kinase inhibitors (e.g., erlotinib (Tarceva, Genentech, Inc.), imatinib (Gleevec, Novartis Pharmaceutical Corporation), gefitinib (Iressa, AstraZeneca Pharmaceuticals), dasatinib (Sprycel, Brystol-Myers Squibb), sunitinib (Sutent, Pfizer, Inc.), nilotinib (Tazigna, Novartis Pharmaceutical Corporation), lapatinib (Tykerb, GlaxoSmithKline Pharmaceuticals), sorafenib (Nexavar, Bayer and Onyx), phosphoinositide 3-kinase (PI3K).

[0796] antimetabolites

[0797] The linker of the present invention may be used to conjugate an antibody to at least one antimetabolite. Antimetabolites are a type of chemotherapy treatment that is very similar to the normal substance within a cell. When a cell incorporates antimetabolite into its cellular metabolism, the result is negative for the cell, and, for example, the cell cannot be isolated. Antimetabolites are classified according to the substance they interfere with. Examples of antimetabolites that may be used in the ADC of the present invention include, but are not limited to, the following: folic acid antagonists ( for example , methotrexate), pyrimidine antagonist ( for example , 5-fluorouracil, foxuridine, cytarabine, capecitabine, and gemcitabine), purine antagonists ( for example , 6-mercaptopurine and 6-thioguanine) and adenosine deaminase inhibitors ( for example , cladribin, fludarabine, nelarabin, and pentostatin), these are described in more detail below.

[0798] Antifoliant

[0799] The linker of the present invention may be used to conjugate an antibody to at least one antifolate agent. Antifolate agents are a subclass of antimetabolites structurally similar to folates. Representative examples include, but are not limited to, the following: methotrexate, 4-amino-folate (also known as aminopterin and 4-aminopteroic acid), lometrexole (LMTX), pemetrexed (Alimpta, Eli Lilly and Company), and trimetrexate (Nutrexin, Ben Venue Laboratories, Inc.).

[0800] purine antagonists

[0801] The linker of the present invention may be used to conjugate an antibody to at least one purine antagonist. Purine analogs are a subclass of antimetabolites that are structurally similar to the group of compounds known as purines. Representative examples of purine antagonists include, but are not limited to: azathioprine (Azasan, Salix; Imuran, GlaxoSmithKline), cladribin (Leustatin [also known as 2-CdA], Janssen Biotech, Inc.), mercaptopurine (Puriunethol [also known as 6-mercaptoethanol], GlaxoSmithKline), fludarabine (Fludara, Genzyme Corporation), pentostatin (Nipent, also known as 2′-deoxycoformycin (DCF)), and 6-thioguanine (Lanvis [also known as thioguanine], GlaxoSmithKline).

[0802] Pyrimidine antagonists

[0803] The linker of the present invention may be used to conjugate an antibody to at least one pyrimidine antagonist. Pyrimidine antagonists are a subclass of antimetabolites structurally similar to the group of compounds known as purines. Representative examples of pyrimidine antagonists are, but not limited to, azacitidine (Vidaza, Celgene Corporation), capecitabine (Xeloda, Roche Laboratories), cytarabine (also known as cytosine arabinoside and arabinocytosine, Bedford Laboratories), decitabine (Dacogen, Eisai Pharmaceuticals), 5-fluorouracil (Adrucil, Teva Pharmaceuticals; Efudex, Valeant Pharmaceuticals, Inc), 5-fluoro-2'-deoxyuridine 5'-phosphate (FdUMP), 5-fluorouridine triphosphate, and gemcitabine (Gemzar, Eli Lilly and Company).

[0804] Boron-containing preparations

[0805] The linker of the present invention may be used to conjugate an antibody to at least one boron-containing formulation. Boron-containing formulations include a class of cancer therapeutic compounds that inhibit cell proliferation. Representative examples of boron-containing formulations include, but are not limited to, the following: borophysin and bortezomib (Velcade, Millenium Pharmaceuticals).

[0806] chemical protective agent

[0807] The linker of the present invention may be used to conjugate an antibody to at least one chemoprotective agent. Chemoprotective drugs are a class of compounds that help protect the body against certain toxic effects of chemotherapy. Chemoprotective agents may be administered with various chemotherapy agents to protect healthy cells from the toxic effects of chemotherapy drugs, while simultaneously allowing cancer cells to be treated with the administered chemotherapy agents. Representative chemoprotective agents include, but not limited to, amifostine (Ethyol, Medimmune, Inc.), which reduces nephrotoxicity associated with cumulative doses of cisplatin; dexrazoxane (Totect, Apricus Pharma; Zinecard), for the treatment of extravasation caused by the administration of anthracycline (Totect) and heart-related complications caused by the administration of the antitumor antibiotic doxorubicin (Zinecard); and Mesna (Mesnex, Bristol-Myers Squibb), which is used to prevent hemorrhagic cystitis during chemotherapy treatment with ipoccarmid.

[0808] Hormonal preparations

[0809] The linker of the present invention may be used to conjugate an antibody to at least one hormone preparation. Hormonal preparations (including synthetic hormones) are compounds that interfere with the production or activity of hormones produced endogenously by the endocrine system. In some embodiments, these compounds interfere with cell growth or produce cytotoxic effects. Non-limiting examples include androgens, estrogens, medroxyprogesterone acetate (Provera, Pfizer, Inc.), and progestins.

[0810] Anti-hormone preparations

[0811] The linker of the present invention may be used to conjugate an antibody to at least one anti-hormone preparation. The "anti-hormone" preparation is a preparation that inhibits the production of a specific endogenous hormone and / or prevents its function. In one embodiment, the anti-hormone preparation inhibits the growth of various cancer cells by interfering with the activity of a hormone selected from the group comprising androgens, estrogens, progesterone, and gonadotropin-releasing hormones. Representative examples of anti-hormonal agents include, but are not limited to, the following: aminoglutethimide, anastrozole (Arimidex, AstraZeneca Pharmaceuticals), bicalutamide (Casodex, AstraZeneca Pharmaceuticals), cyproterone acetate (Cyprostat, Bayer PLC), degarelix (Firmagon, Ferring Pharmaceuticals), exemestane (Aromasin, Pfizer Inc.), flutamide (Drogenil, Schering-Plough Ltd), fulvestrant (Faslodex, AstraZeneca Pharmaceuticals), goserelin (Zolodex, AstraZeneca Pharmaceuticals), letrozole (Femara, Novartis Pharmaceuticals Corporation), leuprolide (Prostap), Lupron, medroxyprogesterone acetate (Provera, Pfizer Inc.), megestrol acetate (Megace, Bristol-Myers Squibb Company), tamoxifen (Nolvadex, AstraZeneca Pharmaceuticals), and triptorelin (Decapetyl, Ferring).

[0812] corticosteroids

[0813] The linker of the present invention may be used to conjugate an antibody to at least one corticosteroid. Corticosteroids may be used in the ADC of the present invention to reduce inflammation. Examples of corticosteroids include, but are not limited to, the following: glucocorticoids, e.g., prednisone (Deltasone, Pharmacia & Upjohn Company, a division of Pfizer, Inc.).

[0814] Photoactive therapeutic agent

[0815] The linker of the present invention may be used to conjugate an antibody to at least one photoactive therapeutic agent. The photoactive therapeutic agent comprises a compound that can be deployed to kill treated cells upon exposure to electromagnetic radiation of a specific wavelength. The therapeutically relevant compound absorbs electromagnetic radiation at a wavelength that penetrates tissue. In a preferred embodiment, the compound is administered in a non-toxic form that can produce a photochemical effect toxic to cells or tissues upon sufficient activation. In another preferred embodiment, these compounds are retained by cancerous tissue and are easily removed from normal tissue. Non-limiting examples include various chromogenic agents and dyes.

[0816] Oligonucleotides

[0817] The linker of the present invention may be used to conjugate an antibody to at least one oligonucleotide. Oligonucleotides are made of short nucleic acid chains that act by interfering with the processing of genetic information. In some embodiments, the oligonucleotides used in the ADC are unmodified single-stranded and / or double-stranded DNA or RNA molecules, while in other embodiments, these therapeutic oligonucleotides are chemically modified single-stranded and / or double-stranded DNA or RNA molecules. In one embodiment, the oligonucleotides used in the ADC are relatively short (19-25 nucleotides) and hybridize to a unique nucleic acid sequence from the total pool of nucleic acid targets present in the cell. Some of the important oligonucleotide technologies include antisense oligonucleotides (including RNA interference (RNAi)), aptamers, CpG oligonucleotides, and ribozymes.

[0818] antisense oligonucleotide

[0819] The linker of the present invention may be used to conjugate an antibody to at least one antisense oligonucleotide. The antisense oligonucleotide is designed to bind to RNA via Watson-Click hybridization. In some embodiments, the antisense oligonucleotide is complementary to a nucleotide encoding region, domain, portion, or segment of the conjugated antibody. In some embodiments, the antisense oligonucleotide comprises about 5 to about 100 nucleotides, about 10 to about 50 nucleotides, about 12 to about 35, and about 18 to about 25 nucleotides.

[0820] There are multiple mechanisms by which oligonucleotides can inhibit the function of RNA when they bind to the target RNA (Crooke ST. (1999). Biochim.Biophys.Acta, 1489, 30-42). The best-characterized antisense mechanism results in the cleavage of targeted RNA by endogenous cellular nucleases, such as RNase H or nucleases associated with RNA interference mechanisms. However, oligonucleotides that inhibit the regulation of target gene expression, such as splicing or translation arrest, by non-catalytic mechanisms can also be potent and selective regulators of gene function.

[0821] Another RNase-dependent antisense mechanism that has recently received much attention is RNAi (Fire et al. (1998). Nature, 391, 806-811; Zamore PD. (2002). Science, 296, 1265-1269). RNA interference (RNAi) is a post-transcriptional process in which double-stranded RNA inhibits gene expression in a sequence-specific manner. In some embodiments, the RNAi effect is achieved through the introduction of a relatively longer double-stranded RNA (dsRNA), while in preferred embodiments, this RNAi effect is achieved by the introduction of a shorter double-stranded RNA, e.g., small interfering RNA (siRNA) and / or microRNA (miRNA). In other embodiments, RNAi can also be achieved by introducing a plasmid that generates dsRNA complementary to the target gene. In each of the aforementioned embodiments, the double-stranded RNA is designed to interfere with the gene expression of a specific target sequence within the cell. Generally, the mechanism involves converting dsRNA into short RNA that directs ribonucleases to homologous mRNA targets (Abstract, Ruvkun, Science 2294:797 (2001)), which degrades the corresponding endogenous mRNA and thereby obtains regulation of gene expression. Notably, dsRNA has been reported to possess antiproliferative properties, which also allows for the expectation of therapeutic applications (Aubel et al., Proc. Natl. Acad. Sci., USA 88:906 (1991)). For example, synthetic dsRNA has been shown to inhibit tumor growth in mice (Levy et al., Proc. Nat. Acad. Sci., USA, 62:357-361 (1969)), and is active in the treatment of leukemic mice (Zeleznick et al., Proc. Soc. Exp. Biol. Med.130:126-128 (1969)), and inhibits chemically induced tumor formation in mouse skin (Gelboin et al., Science 167:205-207 (1970)). Accordingly, in a preferred embodiment, the present invention provided for the use of an antisense oligonucleotide in an ADC for the treatment of breast cancer. In another embodiment, the present invention provides a composition and a method for initiating an antisense oligonucleotide treatment, wherein the dsRNA interferes with the target cell expression of EGFR at the mRNA level. dsRNA refers to naturally occurring RNA, partially purified RNA, recombinant RNA, synthetic RNA, as well as naturally occurring RNA and modified RNA by the encapsulation of non-standard nucleotides, non-nucleotide materials, nucleotide analogs (e.g., lock nucleic acids (LNA)), deoxyribonucleotides, and any combination thereof, as used above. The RNA of the present invention needs to be sufficiently similar to natural RNA having the ability to encode antisense oligonucleotide-based regulation as described herein.

[0822] Aptamer

[0823] The linker of the present invention can be used to conjugate an antibody to at least one aptamer. An aptamer is a nucleic acid molecule selected from a random pool based on its ability to bind to other molecules. Like antibodies, aptamers can bind to target molecules having peculiar affinities and specificities. In many embodiments, the aptamer assumes a complex sequence-dependent, three-dimensional shape that causes it to interact with a target protein, obtains a tightly bound complex similar to antibody-antigen interactions, and thereby interferes with the function of said protein. The specific capacity of an aptamer to bind tightly and specifically to its target protein highlights its potential as a targeted molecular therapy.

[0824] CpG oligonucleotides

[0825] The linker of the present invention can be used to conjugate an antibody to at least one CpG oligonucleotide. Bacterial and viral DNA are known to be potent activators of innate and specific immunity in humans. These immunological properties are associated with unmethylated CpG dinucleotide motifs found in bacterial DNA. Due to the fact that these motifs are rare in humans, the human immune system has evolved the ability to recognize these motifs as early signs of infection and subsequently initiate an immune response. Therefore, oligonucleotides containing these CpG motifs can be pioneered to initiate an anti-tumor immune response.

[0826] Ribozyme

[0827] The linker of the present invention can be used to conjugate an antibody to at least one ribozyme. A ribozyme is a catalytic RNA molecule ranging from about 40 to 155 nucleotides. The ability of a ribozyme to recognize and cleave specific RNA molecules makes it a potential candidate for a therapeutic agent. Representative examples include angiozymes.

[0828] Radionuclide preparations (radioactive isotopes)

[0829] The linker of the present invention can be used to conjugate an antibody to at least one radionuclide preparation. The radionuclide preparation comprises a preparation characterized by an indeterminate nucleus capable of undergoing radioactive decay. The basis for successful radionuclide therapy depends on a sufficient concentration and long-term retention of the radionuclide by cancer cells. Other factors considered include the radionuclide half-life, the energy of the emitted particles, and the maximum range the emitted particles can travel. In a preferred embodiment, the therapeutic agent is a radionuclide selected from the group consisting of: 111In, 177Lu, 212Bi, 213Bi, 211At, 62Cu, 64Cu, 67Cu, 90Y, 125I, 131I, 32P, 33P, 47Sc, 111Ag, 67Ga, 142Pr, 153Sm, 161Tb, 166Dy, 166Ho, 186Re, 188Re, 189Re, 212Pb, 223Ra, 225Ac, 59Fe, 75Se, 77As, 89Sr, 99Mo, 105Rh, 109Pd, 143Pr, 149Pm, 169Er, 194Ir, 198Au, 199Au, and 211Pb. Radionuclides that substantially decay Auger-emitting particles are also preferred. For example, Co-58, Ga-67, Br-80m, Tc-99m, Rh-103m, Pt-109, In-1111, Sb-119, 1-125, Ho-161, Os-189m, and Ir-192. The decay energies of useful beta-particle-emitting nuclides are preferably Dy-152, At-211, Bi-212, Ra-223, Rn-219, Po-215, Bi-211, Ac-225, Fr-221, At-217, Bi-213, and Fm-255. The decay energy of useful alpha-particle-emitting radionuclides is preferably 2,000-10,000 keV, more preferably 3,000-8,000 keV, and most preferably 4,000-7,000 keV.Additional potential radioactive isotopes used are 11C, 13N, 150, 75Br, 198Au, 95Ru, 97Ru, 103Ru, 105Ru, 107Hg, 203Hg, 121mTe, 122mTe, 125mTe, 165Tm, 167Tm, 168Tm, 197Pt, 109Pd, 105Rh, 142Pr, 143Pr, 161Tb, 166Ho, 199Au, 57Co, 58Co, 51Cr, 59Fe, 75Se, 201Tl, 225Ac, 76Br, 169Yb. etc. Includes

[0830] radiosensitizer

[0831] The linker of the present invention may be used to conjugate an antibody to at least one radiosensitizer. The term "radiosensitizer" as used herein is defined as a molecule, preferably a low molecular weight molecule, which is administered to animals in a therapeutically effective dose to increase the resensitization of cells to electromagnetic radiation and / or to promote the treatment of diseases treatable by electromagnetic radiation. A radiosensitizer is a agent that makes cancer cells more sensitive to radiation therapy, while typically having a much smaller effect on normal cells. Thus, a radiosensitizer may be used in combination with a radiolabeled antibody or an ADC. The addition of a radiosensitizer may result in enhanced efficacy compared to treatment with the radiolabeled antibody or antibody fragment alone. Radiosensitizers are described below: DMGoldberg (ed.), Cancer Therapy with Radiolabeled Antibodies, CRC Press (1995). Examples of radiosensitizers include gemcitabine, 5-fluorouracil, taxane, and cisplatin.

[0832] Radiosensitizers can be activated by electromagnetic radiation of X-rays. Representative examples of X-ray-activated radiosensitizers include, but are not limited to, metronidazole, misonidazole, desmethylmisonidazole, pimonidazole, etanidazole, nimorazole, mitomycin C, RSU 1069, SR 4233, E09, RB 6145, nicotinamide, 5-bromodeoxyuridine (BUdR), 5-iododeoxyuridine (IUdR), bromodeoxycitidine, fluorodeoxyuridine (FUdR), hydroxyurea, cisplatin, and their therapeutically effective analogs and derivatives. Alternatively, radiosensitizers can be activated using photodynamic therapy (PDT). Representative examples of photodynamic radiosensitizers include, but are not limited to, the following: hematoporphyrin derivatives, photophrine(r), benzoporphyrin derivatives, NPe6, tin ethioporphyrin (SnET2), pheophorbide a, bacteriochlorophyll a, naphthalocyanine, phthalocyanine, zinc phthalocyanine, and therapeutically effective analogs and derivatives thereof.

[0833] Topoisomerase inhibitor

[0834] The linker of the present invention may be used to conjugate an antibody to at least one topoisomerase inhibitor. Topoisomerase inhibitors are chemotherapeutic agents designed to interfere with the action of topoisomerase enzymes (topoisomerase I and II), which are enzymes that control changes in DNA structure during normal cell cycles by the catalytic, subsequent breakdown, and recombination of the phosphodiester backbone of DNA strands. Representative examples of DNA topoisomerase I inhibitors include, but are not limited to, the following: camptothecin and its derivatives irinotecan (CPT-11, Camptosar, Pfizer, Inc.) and topotecan (Hycamtin, GlaxoSmithKline Pharmaceuticals). Representative examples of DNA topoisomerase II inhibitors include, but are not limited to, the following: amsacrine, daunorubicin, doxorubicin, epipodophyllotoxin, ellipticin, epirubicin, etoposide, razoxic acid, and tenifoside.

[0835] Tyrosine kinase inhibitors

[0836] The linker of the present invention may be used to conjugate an antibody to at least one tyrosine kinase inhibitor. Tyrosine kinase is an intracellular enzyme that functions to attach a phosphate group to the amino acid tyrosine. By blocking the ability of a protein tyrosine kinase to function, tumor growth can be inhibited. Examples of tyrosine kinases that may be used in the ADC of the present invention include, but are not limited to, the following: axitinib, bosutinib, cediranib, dasatinib, erlotinib, gefitinib, imatinib, lapatinib, restaurtinib, nilotinib, cemaxanib, sunitinib, and vandetanib.

[0837] Other preparations

[0838] Examples of other agents that may be used in the ADC of the present invention include, but are not limited to: abrine (e.g., abrine A chain), alpha toxin, Allurites fordi protein, amatoxin, crotin, cursin, diantin protein, diphtheria toxin (e.g., diphtheria A chain and unbound active fragment of diphtheria toxin), deoxyribonuclease (Dnase), gelonin, mitogelin, modexin A chain, Momordica charantia inhibitor, neomycin, onconase, phenomycin, phytolacaramericana protein (PAPI, PAPII, and PAP-S), porkeyweed antiviral protein, Pseudomonas endotoxin, Pseudomonas exotoxin (e.g., exotoxin A chain (Pseudomonas aeruginosa)), restrictocin, lysine A chain, ribonuclease (Rnase), Sapaonaria officinalis inhibitor, saporin, Alpha-sarcin, Staphylococcus enterotoxin-A, tetanustoxin, cisplatin, carboplatin, and oxaliplatin (Eloxatin, Sanofi Aventis), proteasome inhibitors (e.g., PS-341 [bortezomib or Velcade]), HDAC inhibitors (vorinostat (Zolinza, Merck & Company, Inc.)), velinostat, entinostat, moschotinostat, and panobinostat), COX-2 inhibitors, substituted ureas, heat shock protein inhibitors (e.g., zeldanamycin and its numerous analogs), adrenocorticosteroids, and trichothecenes (e.g., see WO 93 / 21232). Other preparations also include: asparaginase (Espar, Lundbeck Inc.), hydroxyurea, levamisole, mitotan (Lysodren, Bristol-Myers Squibb), and tretinoin (Renova, Valeant Pharmaceuticals Inc.).

[0839] It should be noted that the aforementioned groups of the drug moiety that can be used in the ADC of the present invention are not exclusive, given that specific examples of drugs may be found in one or more categories, for example, that ansamitocine is both a mitotic inhibitor and an antitumor antibiotic.

[0840] All stereoisomers of the above drug moiety are considered as any combination of the R and S arrangement forms at the chiral carbon of D of the compound of the present invention.

[0841] "Detectable moiety" or "marker" refers to a composition detectable by spectroscopic, photochemical, biochemical, immunochemical, radiological, or chemical means. For example, useful labels include the following: 32 P, 35 S, fluorescent dye, electron-densifying reagent, enzyme ( for example , enzymes commonly used in ELISA), biotin-streptavidin, deoxygenin, hapten, and proteins for which antiserum or monoclonal antibodies are available, or nucleic acid molecules having sequences complementary to the target. Detectable moiety is often a measurable signal, for example It generates a radioactive signal, a color signal, or a fluorescent signal, which can be used to quantify the amount of detectable moiety bound in a sample. Quantification of the signal can be achieved, for example, by scintillation counting, density gauge, flow cell analysis, ELISA, or direct analysis by mass spectrometry of the original or subsequently digested peptide (one or more peptides may be tested). Those skilled in the art are familiar with techniques and detection means for the labeled compound of interest. These techniques and methods are known and are well known in the field.

[0842] A detection probe refers to the following: (i) a substance capable of providing a detectable signal; (ii) a substance capable of interacting with a first probe or a second probe to modify the detectable signal provided by, for example, fluorescence resonance energy transfer (FRET); (iii) a substance capable of stabilizing an interaction with an antigen or ligand or increasing binding affinity; (iv) physical parameters, for example, charge, hydrophobicity, etc. A substance that can affect electromobility or cell-invasive action, or (v) a substance that can modulate ligand affinity, antigen-antibody binding, or ion complex formation.

[0843] In certain embodiments, FRET technology may be used to distinguish between a molecule exposed to conditions that activate a trigger group and an intact molecule, for example, by attaching an acceptor chromophore to a central Ar ring and attaching the acceptor chromophore to Q.

[0844] In some embodiments, the present invention provides for the use of compounds disclosed as imaging agents (e.g., fluorescent groups or chelators), such as fluorescein, rhodamine, lanthanide phosphors, and derivatives thereof. Examples of fluorophores include, but are not limited to, fluorescein isothiocyanate (FITC) (e.g., 5-FITC), fluorescein amidite (FAM) (e.g., 5-FAM), eosin, carboxyfluorescein, erythrosine, Alexa Fluor.RTM (e.g., Alexa 350, 405, 430, 488, 500, 514, 532, 546, 555, 568, 594, 610, 633, 647, 660, 680, 700, or 750), carboxytetramethylrhodamine (TAMRA) (e.g., 5-TAMRA), tetramethylrhodamine (TMR), and sulforodamine (SR) (e.g., SR101). .Examples of chelators include, but are not limited to, 1,4,7,10-tetraazcyclododecane-N,N',N",N"'-tetraacetic acid (DOTA), 1,4,7-triazcyclononan-1,4,7-triacetic acid (NOTA), 1,4,7-triazcyclononan, 1-glutaric acid-4,7-acetic acid (NODAGA), diethylenetriaminepentaacetic acid (DTPA), and 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA).

[0845] antibodies

[0846] The antibody of the ADC may be any antibody that typically binds to an antigen expressed on the surface of a target cell of interest, but is not specifically required to be so. The antigen may, in some embodiments, internalize the ADC bound thereto into the cell, though it is not necessary. The target cell of interest may include a cell in which induction of apoptosis is desired. The target antigen may be any protein, glycoprotein, polysaccharide, lipoprotein, expressed on the target cell of interest. etc. It may be, but typically, a protein that is uniquely expressed on either a target cell other than a normal or healthy cell, or is overexpressed on a target cell compared to a normal or healthy cell, thereby allowing the ADC to selectively target specific cells of interest, e.g., tumor cells. As will be known to those skilled in the art, the specific antigen and the antibody selected accordingly will depend on the identity of the desired target cell of interest. In a specific embodiment, the antibody of the ADC is an antibody suitable for administration to humans.

[0847] Antibodies (Abs) and immunoglobulins (Igs) are glycoproteins with identical structural characteristics. While antibodies exhibit binding specificity to specific targets, immunoglobulins include both antibodies and other antibody-like molecules that lack target specificity. Natural antibodies and immunoglobulins are generally heterotemeric glycoproteins of approximately 150,000 daltons, composed of two identical light chains (L) and two identical heavy chains (H). Each heavy chain has a variable domain (VH) at one end followed by numerous constant domains. Each light chain has a variable domain (VL) at one end and a constant domain at its other end.

[0848] The reference "VH" refers to a variable region of the immunoglobulin heavy chain of an antibody, comprising the heavy chain of Fv, scFv, or Fab. The reference "VL" refers to a variable region of the immunoglobulin light chain, comprising the light chain of Fv, scFv, dsFv, or Fab.

[0849] In this specification, the term “antibody” is used in the broadest sense and refers to an immunoglobulin molecule that specifically binds to or reacts immunologically with a specific antigen, and is not limited to Murin, chimeric antibodies, humanized antibodies, heteroconjugate antibodies ( for example Includes polyclonal, monoclonal, genetically engineered, and otherwise modified forms of antibodies, including dispecific antibodies, diabadies, triabadies, and tetraabadies), and antigen-binding fragments of antibodies, such as Fab', F(ab')2, Fab, Fv, rIgG, and scFv fragments. The term "scFv" refers to a single-chain Fv antibody in which the variable domains of the heavy chain and light chain from a traditional antibody are linked to form a single chain.

[0850] Antibodies can be rodents, humans, humanized, chimeras, or derived from other species. Antibodies are proteins produced by the immune system capable of recognizing and binding to specific antigens. (Janeway, C., Travers, P., Walport, M., Shlomchik (2001)) Immuno Biology , 5th Ed., Garland Publishing, New York). Target antigens generally have numerous binding sites, also known as epitopes, recognized by CDRs on multiple antibodies. Each antibody that specifically binds to a different epitope has a different structure. Therefore, a single antigen may have one or more corresponding antibodies. Antibodies are full-length immunoglobulin molecules or the immunologically active portion of a full-length immunoglobulin molecule, in other words , comprising a molecule containing an antigen-binding site that immunospecifically binds to an antigen or a part thereof of a target of interest, such target comprises, without limitation, cells or cancer cells that produce autoimmune antibodies associated with autoimmune diseases. The immunoglobulins disclosed herein are of any type ( for example , IgG, IgE, IgM, IgD, and IgA), class ( for example It may be a subclass of immunoglobulin molecules (i.e., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2). Immunoglobulins may be derived from any species. In one embodiment, however, immunoglobulins are of human, rodent, or rabbit origin.

[0851] The term "antibody fragment" refers to a portion of a full-length antibody, generally the target binding or variable region. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments. The "Fv" fragment is a minimal antibody fragment containing a complete target recognition and binding site. This region consists of a dimer of one heavy chain and one light chain variable domain (VH-VL dimer) bound by strong, non-covalent bonds. In this configuration, three CDRs of each variable domain interact to define the target binding site on the surface of the VH-VL dimer. Often, six CDRs confer target binding specificity to the antibody. However, in some cases, even a single variable domain (or half of the Fv containing only three CDRs specific to the target) may possess the ability to recognize and bind to the target. "Single-chain Fv" or "scFv" antibody fragments contain the antibody's VH and VL domains in a single polypeptide chain. Generally, the Fv polypeptide comprises a polypeptide linker between the VH and VL domains that enables the scFv to form a desired structure for additional target binding. The "single-domain antibody" consists of a single VH or VL domain that exhibits sufficient affinity for the target. In a specific embodiment, the single-domain antibody is a camelized antibody ( for example , Riechmann, 1999, Journal of Immunological Methods 231:25-38).

[0852] The Fab fragment contains a constant domain of the light chain and a first constant domain (CH1) of the heavy chain. The Fab' fragment differs from the Fab fragment by the addition of a few residues at the carboxyl terminus of the heavy chain CH1 domain, which contains one or more cystes from the antibody hinge region. The F(ab') fragment is produced by the cleavage of a disulfide bond at the hinge cysteine ​​of the F(ab')2 pepsin digestion product. Further chemical coupling of antibody fragments is known to those skilled in the art.

[0853] Both the light and heavy chain variable domains possess a complementary determining region (CDR), also known as the supervariability region. The more highly conserved portion of the variable domain is called the framework (FR). As is known in the art, the amino acid positions / boundaries marking the supervariability region of an antibody may vary depending on the context and various definitions known in the art. Within the variable domain, some positions may be observed as hybrid supervariability regions, in that these positions may be considered outside the supervariability region under a different set of criteria, while being considered within the supervariability region under one set of criteria. One or more of these positions may also be found in the extended supervariability region. In each chain, the CDR is held together in very close proximity by the FR region and contributes to the formation of the antibody's target binding site along with the CDR from the other chain (Reference: Kabat etc. , Sequences of Proteins of Immunological Interest (National Institute of Health, Bethesda, Md. 1987). As used herein, the numbering of immunoglobulin amino acid residues is, unless otherwise indicated, Kabat etc. It is carried out according to the immunoglobulin amino acid residue numbering system.

[0854] In certain embodiments, the antibody of the ADC of the present disclosure is a monoclonal antibody. The term " monoclonal "Antibody" (mAb) is not a method of production, for example ...refers to antibodies derived from a single copy or clone, including any eukaryotic, prokaryotic, or phage clone. Preferably, the monoclonal antibodies of this disclosure exist in a homogeneous or substantially homogeneous population. Monoclonal antibodies comprise both the intact molecule capable of specifically binding to a protein, as well as antibody fragments (e.g., Fab and F(ab')2 fragments). The Fab and F(ab')2 fragments lack the Fc fragment of the intact antibody, are cleared more rapidly from animal circulation, and may have less non-specific tissue binding than the intact antibody (Wahl etc. , 1983, J. Nucl.Med 24:316). Monoclonal antibodies useful for the present disclosure may be produced using various techniques known in the art, including the use of hybridoma, recombinant, and phage display techniques, or combinations thereof. The antibodies of the present disclosure include chimeric, primate-modified, humanized, or human antibodies.

[0855] In most cases, antibodies consist solely of genetically encoded amino acids, whereas in some embodiments, non-encoded amino acids may be specifically incorporated. Examples of non-encoded amino acids that may be incorporated into antibodies for use in stoichiometry and attachment site control, as well as methods for preparing such modified antibodies, are discussed below: Tian etc. , 2014, Proc Nat'l Acad Sci USA 111(5):1766-1771 and Axup etc. , 2012, Proc Nat'l Acad Sci USA 109(40):16101-16106 (the whole contents of which are incorporated herein by reference).

[0856] In certain embodiments, the antibody of the ADC described herein is a chimeric antibody. As used herein, the term “chimeric” antibody refers to a non-human immunoglobulin, such as a rat or mouse antibody, typically selected from a human immunoglobulin template, and an antibody having a variable sequence derived from a human immunoglobulin constant region. Methods for producing chimeric antibodies are known in the art. for example , Morrison, 1985, Science 229(4719):1202-7; Oi etc. , 1986, BioTechniques 4:214-221; Gillies etc. , 1985, J. Immunol . Methods 125:191-202; U.S. Patent Nos. 5,807,715; 4,816,567; and 4,816,397 (the entirety of which is incorporated herein by reference).

[0857] In certain embodiments, the antibody of the ADC described herein is a humanized antibody. Non-human ( for example The "humanized" form of the antibody is a chimeric immunoglobulin, immunoglobulin chain, or fragment thereof (e.g., the Fv, Fab, Fab', F(ab')2, or other target-binding subdomains of the antibody) containing a minimal sequence derived from a non-human immunoglobulin. Generally, the humanized antibody will comprise at least one, and typically two, variable domains substantially all, wherein all or substantially all CDR regions correspond to those of the non-human immunoglobulin and all or substantially all FR regions correspond to those of the human immunoglobulin sequence. The humanized antibody may also comprise at least a portion of the immunoglobulin constant region (Fc), typically of the human immunoglobulin common sequence. Methods of antibody humanization are known in the art. for example , Riechmann etc. , 1988, Nature 332:323-7; U.S. Patent Nos. 5,530,101; 5,585,089; 5,693,761; 5,693,762; and U.S. Patent No. 6,180,370, Queen etc. ;EP239400; PCT Publication WO 91 / 09967; U.S. Patent No. 5,225,539; EP592106; EP519596; Padlan, 1991, Mol . Immunol . , 28:489-498; Studnicka etc. , 1994, Prot . Eng . 7:805-814; Roguska etc. , 1994, Proc . Natl . Acad Sci USA See 91:969-973; and U.S. Patent No. 5,565,332 (all of which are incorporated herein by reference in their entirety).

[0858] In certain embodiments, the antibody of the ADC described herein is a human antibody. An entirely “human” antibody may be desirable for the therapeutic treatment of human patients. As used herein, “human antibody” comprises an antibody having the amino acid sequence of a human immunoglobulin and comprises an antibody isolated from an animal transgenic to one or more human immunoglobulins or from a human immunoglobulin library and not expressing the endogenous immunoglobulin. The human antibody may be implemented by various methods known in the art, including a phage display method using an antibody library derived from a human immunoglobulin sequence. U.S. Patent Nos. 4,444,887, 4,716,111, 6,114,598, 6,207,418, 6,235,883, 7,227,002, 8,809,151 and U.S. Application release See No. 2013 / 189218 (the contents thereof are incorporated herein by reference in their entirety). Human antibodies may also be produced using transgenic mice that cannot express functional endogenous immunoglobulins but can express human immunoglobulin genes. for example, see U.S. Patent Nos. 5,413,923; 5,625,126; 5,633,425; 5,569,825; 5,661,016; 5,545,806; 5,814,318; 5,885,793; 5,916,771; 5,939,598; 7,723,270; 8,809,051 and U.S. Publication No. 2013 / 117871 (the entirety of which is incorporated herein by reference). Additionally, companies such as Medarex (Princeton, NJ), Astellas Pharma (Deerfield, Ill.), and Regeneron (Tarrytown, NY) may provide human antibodies to selected antigens using technology similar to that described above. Fully human antibodies recognizing selected epitopes can be generated using a technique referred to as "induced selection." Non-human monoclonal antibodies selected in this approach, for example Mouse antibodies are used to guide the selection of fully human antibodies that recognize the same epitope (Jespers etc. , 1988, Biotechnology 12:899-903).

[0859] In certain embodiments, the antibody of the ADC described herein is a primate-modified antibody. The term "primate-modified antibody" refers to an antibody comprising a monkey variable region and a human constant region. Methods for producing primate-modified antibodies are known in the art. for example , see U.S. Patent Nos. 5,658,570; 5,681,722; and 5,693,780 (the entirety of which is incorporated herein by reference).

[0860] In certain embodiments, the antibodies of the ADCs described herein are bispecific antibodies or bivariable domain antibodies (DVDs). Bispecific and DVD antibodies are monoclonal, often human or humanized, antibodies having binding specificity to at least two different antigens. DVDs are described, for example, in U.S. Patent No. 7,612,181 (the disclosure thereof is incorporated herein by reference).

[0861] In certain embodiments, the antibodies of the ADCs described herein are derived antibodies. For example, but not limited to, derived antibodies are typically modified by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protective / blocking groups, proteolytic cleavage, linkage to cellular ligands or other proteins, etc. Any numerous chemical modifications, but not limited to, specific chemical cleavage, acetylation, formylation, metabolic synthesis of tunicamycin, etc. It can be performed by known techniques including. Additionally, derivatives, for example , using Ambrix technology, it may contain one or more non-natural amino acids ( for example , Wolfson, 2006, Chem.Biol. See 13(10):1011-2).

[0862] In certain embodiments, the antibody of the ADC described herein has a modified sequence to alter at least one constant region-mediated biological effector function relative to the corresponding wild-type sequence. For example, in some embodiments, the antibody has at least one constant region-mediated biological effector function relative to the unmodified antibody, for example , can be modified to reduce reduced binding to the Fc receptor (FcR). FcR binding can be reduced by mutating the immunoglobulin constant region segment of the antibody in the specific region required for FcR interaction ( for example, Canfield and Morrison, 1991, J. Exp . Med 173:1483-1491; and Lund etc. , 1991, J. Immunol. See 147:2657-2662).

[0863] In a specific embodiment, the antibody of the ADC described herein acquires or improves at least one invariant region-mediated biological effector function compared to an unmodified antibody, for example , is modified to enhance FcγR interaction ( reference , for example , US 2006 / 0134709). For example, antibodies having a constant region that binds to FcγRIIA, FcγRIIB and / or FcγRIIIA with a greater affinity than the corresponding wild-type constant region can be produced according to the methods described herein.

[0864] In some specific embodiments, the antibodies of the ADCs described herein are antibodies that bind to tumor cells, such as cell surface receptors or tumor-associated antigens (TAAs). To discover effective cell targets for cancer diagnosis and therapy, researchers have sought to identify transmembrane or otherwise tumor-associated polypeptides that are specifically expressed on the surface of one or more specific types of cancer cells compared to one or more normal non-cancerous cell(s). Often, such tumor-associated polypeptides are expressed more abundantly on the surface of cancer cells compared to the surface of non-cancerous cells. Such cell surface receptors and tumor-associated antigens are known in the art and can be prepared for use in antibody generation using methods and information known in the art.

[0865] Exemplary cell surface receptors and TAA

[0866] Examples of cell surface receptors and TAAs to which the antibodies of the ADCs described herein may be targeted include, but are not limited to, the various receptors and TAAs listed in Table 1 below. For convenience, information regarding all of these antigens known in the art is listed below and includes the name, alternative name, gene bank deposit number, and primary reference(s) in accordance with the nucleic acid and protein sequence identification protocols of the National Center for Biotechnology Information (NCBI). Nucleic acid and protein sequences corresponding to the listed cell surface receptors and TAAs are available in public databases, such as gene banks.

[0867]

[0868]

[0869]

[0870]

[0871] Exemplary antibody

[0872] Exemplary antibodies to be used with the ADC of the present disclosure include, but are not limited to: 3F8 (GD2), avagovomab (CA-125 (misc)), adecatumumab (EpCAM), aputuzumab (CD20), alacizumab pegol (VEGFR2), ALD518 (IL-6), alemtuzumab (CD52), altumomab pentetate (CEA), amatouximab (mesothelin), anatumonab maphenatox (TAG-72), apolizumab (HLA-DR), arcitumomab (CEA), babituximab (phosphatidylserine), vectumumab (CD22), belimumab (BAFF), becilesomab (CEA-associated antigen), bevacizumab (VEGF-A), vibatuzumab mertansine (CD44 v6), blinatumomab (CD19), brentuximab vedotin ((CD30 (TNFRSF8)), cantuzumab mertansine (mucin CanAg), cantuzumab ravtansine (MUC1), capromab pendetide (prostate carcinoma cells), charlumab (MCP-1), catumaxomab (EpCAM, CD3), CC49 (Tag-72), cBR96-DOX ADC (Lewis-Y antigen), cetuximab (EGFR), citatuzumab vogatox (EpCAM), sixutumab (IGF-1 receptor), clivatuzumab tetraxetane (MUC1), conatumumab (TRAIL-E2), dasetuzumab (CD40), dalotuzumab (insulin-like growth factor 1 receptor), daratumumab ((CD38 (cyclic ADP ribose hydrolase)), dempizumab (DLL4), denosumab (RANKL), detumomab (B-lymphoma Cell), drogitumab (DR5), ducigitumab (ILGF2), ecromeximab (D3 ganglioside), eculizumab (C5), edrecolomab (EpCAM), elotuzumab (SLAMF7), elcilimomab (IL-6), enavatuzumab (TWEAK receptor), enoticumab (DLL4), encituximab (5AC), epitumomab situxetane (episialin), efratuzumab (CD22), ertumaxomab ((HER2 / neu, CD3)), etansizumab (integrin αvβ3), parletuzumab (folic acid receptor 1),FBTA05 (CD20), piclatuzumab (HGF), pizitumab (IGF-1 receptor), flambotumab (TYRP1 (glycoprotein 75)), presolimumab (TGF-1), galiximab (CD80), ganitumab (IGF-I), gemtuzumab ozogamicin (CD33), gilentuximab (carbonic anhydrous enzyme 9 (CA-IX)), glembatumab vedotin (GPNMB), ibritumomab tiuxetane (CD20), icrucumab (VEGFR-1), igobomab (CA-125), IMAB362 (CLDN18.2), imgatuzumab (EGFR), indatuximab lavatansine (SDC1), intertumumab (CD51), inotuzumab ozogamicin (CD22), ipilimumab (CD152), iratumumab ((CD30 (TNFRSF8)), Labetuzumab (CEA), Lambrolizumab (PDCD1), Lexatumumab (TRAIL-R2), Lintuzumab (CD33), Lorvotuzumab Mertansine (CD56), Lucatumumab (CD40), Lumiliximab ((CD23 (IgE receptor)), Mapatumumab (TRAIL-R1), Margetuximab (ch4DS), Matuzumab (EGFR), Milatuzumab (CD74), Mitumomab (GD3 ganglioside), Mogamulizumab (CCR4), Moxetumumab Fasudox (CD22), Nacolomab Tafenatox (C2-42 antigen), Naptumomab Estafenatox (5T4), Narnatumab (RON), Natalizumab (Integrin α4), Necitumumab (EGFR), Nesvacumab (Angiopoietin 2), nimotuzumab (EGFR), nivolumab (IgG4), ocaratuzumab (CD20), ofatumumab (CD20), olaratumab (PDGF-R α), onatuzumab (human scatter detector factor receptor kinase), ontuxizumab (TEM1), ofportuzumab monato (EpCAM), oregovomab (CA-125), othletuzumab (CD37), panitumumab (EGFR), pancomab (tumor-specific glycosylation of MUC1), parsatuzumab (EGFL7), patritumab (HER3), femtumumab (MUC1), pertuzumab (HER2 / neu), fidilizumab (PD-1), finatuzumab vedotin (CD22),Pritumumab (Vimentin), Lacotumumab (N-glycolylneuraminic acid), Radretumab (Fibronectin additional domain-B), Ramucirumab (VEGFR2), Rilotumumab (HGF), Rituximab (CD20), Lovatumumab (IGF-1 receptor), Samalizumab (CD200), Satumomab pendetide (TAG-72), Serivantumab (ERBB3), Sibrotuzumab (FAP), SGN-CD19A (CD19), SGN-CD33A (CD33), Siltuximab (IL-6), Solitomab (EpCAM), Sonefizumab (Sphingosine-1-phosphate), Tavalumab (BAFF), Tacatuzumab tetraxetane (alpha-fetoprotein), Taplitumomab poptox (CD19), Tenatumomab (Thenassine C), Teprotumumab (CD221), TGN1412 (CD28), tilcilimumab (CTLA-4), tigatuzumab (TRAIL-R2), TNX-650 (IL-13), tovetumab (CD40a), trastuzumab (HER2 / neu), TRBS07 (GD2), tremelimumab (CTLA-4), tucotuzumab selmolukin (EpCAM), ublituximab (MS4A), urelumab (4-1BB), vandetanib (VEGF), vantictumab (Prizzled receptor), voloxiximab (integrin α5β1), borsetuzumab mafodotin (CD70), botumumab (tumor antigen CTAA16.88), zalutumumab (EGFR), zanolimumab (CD4), and zatuximab (HER1).

[0873] Method for manufacturing antibodies

[0874] Antibodies of ADCs can be produced by the recombinant expression of immunoglobulin light and heavy chain genes within host cells. For example, to recombinantly express antibodies, host cells are transfected with one or more recombinant expression vectors carrying DNA fragments encoding the antibody's immunoglobulin light and heavy chains, thereby expressing the light and heavy chains in the host cells and, optionally, secreting them into the culture medium in which the host cells are cultured, from which the antibodies can be recovered. Standard recombinant DNA methodologies are used to obtain antibody heavy and light chain genes, to incorporate these genes into a recombinant expression vector, and to introduce the vector into host cells, as described below: Molecular Cloning; A Laboratory Manual, Second Edition (Sambrook, Fritsch and Maniatis (eds), Cold Spring Harbor, NY, 1989), Current Protocols in Molecular Biology (Ausubel, FM etc. , eds., Greene Publishing Associates, 1989) and U.S. Patent No. 4,816,397.

[0875] In one embodiment, the Fc variant antibody is similar to its wild-type equivalent except for a change in its Fc domain. To produce a nucleic acid encoding such an Fc variant antibody, a DNA fragment encoding the Fc domain or a portion of the Fc domain of the wild-type antibody (referred to as the “wild-type Fc domain”) can be synthesized and used as a mutagenic template to produce the antibody as described herein using routine mutagenic techniques; alternatively, the DNA fragment encoding the antibody can be synthesized directly.

[0876] Once DNA fragments encoding a wild-type Fc domain are obtained, these DNA fragments can be further manipulated by standard recombinant DNA techniques, for example, to convert a constant region gene into a full-length antibody chain gene. In these manipulations, the CH-encoding DNA fragment is operably linked to another DNA fragment encoding another protein, such as an antibody variable region or a flexible linker. As used in this context, the term "operably linked" is intended to mean that the two DNA fragments are linked so that the amino acid sequence encoded by the two DNA fragments remains in-frame.

[0877] To express Fc variant antibodies, DNAs encoding partial or full-length light and heavy chains obtained as described above are inserted into an expression vector, thereby operably linking the gene to transcription and translation control sequences. In this context, the term " "Operatorily ligated" is intended to mean that the antibody gene is ligated into a vector, thereby allowing the transcription and translation control sequences within the vector to perform their intended functions of regulating the transcription and translation of the antibody gene. The expression vector and the expression control sequences are selected to be compatible with the expression host cell used. The variant antibody light chain gene and the antibody heavy chain gene may be inserted into separate vectors, or, more typically, both genes are inserted into the same expression vector.

[0878] Antibody genes are placed in the expression vector using the standard method ( for example, is inserted via ligation of a complementary restriction site on the antibody gene fragment and vector, or via smooth end ligation if no restriction site is present. Prior to the insertion of the variant Fc domain sequence, the expression vector may already carry the antibody variable region sequence. Additionally or alternatively, the recombinant expression vector may encode a signal peptide that promotes the secretion of the antibody chain from the host cell. The antibody chain gene may be cloned into the vector so that the signal peptide is inframe-linked to the amino terminus of the antibody chain gene. The signal peptide is an immunoglobulin signal peptide or a heterogeneous signal peptide ( in other words , it can be a signal peptide from a non-immunoglobulin protein.

[0879] In addition to the antibody chain gene, the recombinant expression vector carries a regulatory sequence that controls the expression of the antibody chain gene in the host cell. The term "regulatory sequence" refers to promoters, enhancers, and other expression control elements that control the transcription or translation of the antibody chain gene ( for example It is intended to include polyadenylation signals). Such regulatory sequences are, for example, described below: Goeddel, Gene Expression Technology: Methods in Enzymology 185 (Academic Press, San Diego, Calif., 1990). It will be acknowledged by those skilled in the art that the design of an expression vector, including the selection of regulatory sequences, may depend on factors such as the selection of the host cell to be converted, the expression level of the desired protein, etc. Regulatory sequences suitable for mammalian host cell expression may be promoters and / or enhancers derived from viral elements that induce high levels of protein expression within mammalian cells, such as cytomegalovirus (CMV) (e.g., CMV promoter / enhancer), primate virus 40 (SV40) (e.g., SV40 promoter / enhancer), adenovirus, ( for example, includes the adenovirus major late promoter (AdMLP) and polyoma. For further description of the viral regulatory factors and their sequences, for example , U.S. Patent No. 5,168,062 Stinski, U.S. Patent No. 4,510,245 Bell etc. Author, and U.S. Patent No. 4,968,615 Schaffner etc. Refer to me.

[0880] In addition to the antibody chain gene and regulatory sequence, the recombinant expression vector includes additional sequences, e.g., host cell ( for example , origin of replication) and selectable marker genes can carry sequences that regulate vector replication. Selectable marker genes facilitate the selection of host cells into which the vector is introduced ( for example , U.S. Patent Nos. 4,399,216, 4,634,665 and 5,179,017, all Axel etc. (See reference to [category]). For example, typically selectable marker genes confer resistance to drugs, such as G418, puromycin, blasticidin, hygromycin, or methotrexate, on host cells into which the vector has been introduced. Suitable selectable marker genes include the dihydrofolate reductase (DHFR) gene (for use in DHFR host cells with methotrexate selection / amplification) and the neo gene (for G418 selection). For the expression of light and heavy chains, the expression vector(s) encoding the heavy and light chains are transfected into host cells using standard techniques. Various forms of terminology " "Transfection" refers to various techniques commonly used for the introduction of exogenous DNA into prokaryotic or eukaryotic host cells, for example It is intended to cover electroporation, lipofection, calcium-phosphate precipitation, DEAE-dextran transfection, and other homologous methods.

[0881] It is possible to express antibodies in either a prokaryotic or eukaryotic host cell. In a specific embodiment, antibody expression is performed in a eukaryotic cell, for optimal secretion of a properly folded and immunologically active antibody, for example , is performed in mammalian host cells. An exemplary mammalian host cell for recombinant antibody expression is (Urlaub and Chasin, 1980, Proc. Natl. Acad. Sci. USA Listed in 77:4216-4220, for example As described in Kaufman and Sharp, 1982, Mol. Biol. 159:601-621, the DHFR screening marker used includes Chinese hamster ovaries (CHO cells), NS0 myeloma cells, COS cells, 293 cells, and SP2 / 0 cells. When a recombinant expression vector encoding an antibody gene is introduced into mammalian host cells, the antibody is produced by culturing the host cells for a period sufficient to allow for antibody expression in the host cells or secretion of the antibody into the culture medium in which the host cells are grown. The antibody can be recovered from the culture medium using standard protein purification methods. Host cells can also be used to produce intact antibodies, such as Fab fragments or parts of scFv molecules.

[0882] In some embodiments, the antibody of the ADC may be a bifunctional antibody. Such an antibody, in which one heavy chain and one light chain are specific to one antigen and the other heavy chain and light chain are specific to a second antigen, may be produced by cross-linking the antibody to the second antibody using a standard chemical cross-linking method. A bifunctional antibody may also be produced by expressing a nucleic acid engineered to encode the bifunctional antibody.

[0883] In a specific embodiment, a dual-specific antibody, in other wordsAntibodies that bind one antigen and a second, unrelated antigen using the same binding site can be produced by mutating amino acid residues in the light chain and / or heavy chain CDR. An exemplary second antigen includes a pro-inflammatory cytokine (e.g., lymphotoxin, interferon-γ, or interleukin-1). The bispecific antibody is, for example , can be produced by mutating amino acid residues around the antigen binding site ( for example , Bostrom etc. , 2009, Science (See 323:1610-1614). Bifunctional antibodies can be produced by expressing nucleic acids engineered to encode bispecific antibodies.

[0884] Antibodies are also produced by chemical synthesis ( for example , Solid Phase Peptide Synthesis It can be produced by the method described in The Pierce Chemical Co., Rockford, Ill., , 2nd ed., 1984. Antibodies can also be produced using a cell-free platform ( for example , Chu etc. , Biochemia No. 2, 2001 (Roche Molecular Biologicals).

[0885] The method for recombinant expression of the Fc fusion protein is described below: Flanagan etc. , Methods in Molecular Biolog , vol. 378:Monoclonal Antibodies:Methods and Protocols.

[0886] Once the antibody has been produced by recombinant expression, for the purification of the immunoglobulin molecule, any method known in the art, for example, chromatography ( for exampleProteins can be purified by ion exchange, affinity, particularly by affinity for antigens after selection of protein A or protein G, and by sizing column chromatography), centrifugation, differential solubility, or any other standard technique for protein purification.

[0887] Once separated, the antibody, if desired, for example , high-performance liquid chromatography ( reference , for example , Fisher, Laboratory Techniques In Biochemistry And Molecular Biology It can be further purified by gel filtration chromatography on a Superdex™ 75 column (Pharmacia Biotech AB, Uppsala, Sweden) (Work and Burdon, eds., Elsevier, 1980).

[0888] Imaging Compounds and Sensors

[0889] In specific embodiments, the use of the compound disclosed in the imaging composition and as a sensor is provided herein.

[0890] The sensor may be a biosensor, a chemical sensor, or a molecular switch. A biosensor can confirm the presence or amount of a specific substance by reacting a bio-receptor (a part designed to react with and adsorb biocompatible materials, such as DNA, RNA, antibodies, enzyme proteins, cells, biological membranes, hormone receptors, etc.) having selective specificity with a specific substance (e.g., cancer cells, viruses, various chemicals, etc.), and can perform measurements using a signal transducer (a device that converts the reaction between the specific substance and the biological receptor into an electrical signal using various methods), and may be used in the following applications: medical, environmental, processing industries, military (chemical warfare), research, food, etc. ( for example , Biosensors and Bioelectronics , 2016, 32-45; Pol .J. Environ.Stud.2015, 19-25; Analytica Chimica Acta 568 (2006) 200-210; Biosensors and Bioelectronics 2017, 217-231; ACS Appl. Mater. Interfaces 2015, 7, 20190-20199; Journal of Coastal Life Medicine 2016; 4(3):200-202; Artificial Cells, Blood Substitutes, and Biotechnology , 39:281-288; Journal of Controlled Release 159 (2012) See 154-163).

[0891] Chemical sensors rapidly and accurately monitor specific substances in many fields, such as clinical diagnosis, medical research, chemical measurement, and environmental measurement, by utilizing electrical properties such as electricity, resistance, potential difference, etc., and optical properties such as color, fluorescence, etc., and include the following: gas sensors (hydrogen, oxygen, carbon monoxide), ion sensors (cations, anions, gas-sensitive ions), component sensors (vapor phase, liquid phase, luminescent components), humidity sensors (relative humidity, absolute humidity, condensation), and dust / soot sensors (suspended dust, fine dust, soot, turbidity). etc. ( for example , Chem . Soc Rev. , 2015, 44, 3358; Journal of the Korean Chemical Society , 2010, 451-459; Chem . Sci . , 2015, 6, 1150-1158; KR 10-1549347; J. Phys. Chem. B 2016, 120, 70537061; ACS Appl. Mater. Interfaces 2015, 7, 704712; J. Am. Chem.Soc. 2011, 133, 10960-10965; J. Am. Chem. Soc. 2012, 134, 20412-20420; Org. Lett. 2014, 16, 16801683; J. Org. Chem. 2013, 78, 702705; J. Org. Chem. 2015, 80, 1212912136; ACS Macro Lett. 2014, 3, 1191-1195; New J. Chem. , 2012, 36, 386-393; Chem. Commun. , 2010, 46, 6575-6577; see 2013).

[0892] A molecular switch is a molecule capable of reversibly switching between two or more stable states. Molecules can switch between states in response to changes in environmental stimuli, such as the chemical environment (e.g., pH), light irradiation (e.g., light of a specific wavelength), temperature, electric current, the microenvironment, or the presence of ligands. In some cases, switching between states may depend on a combination of stimuli. The oldest form of a synthetic molecular switch is the pH indicator, which displays distinct colors as a function of pH. Synthetic molecular switches can be applied in molecular computers or reactive drug delivery systems. Molecular switches are also important in biology because many biological functions are based on them, such as allosteric regulation and vision.

[0893] Such biosensors, chemical sensors, and molecular switches may additionally include additional photoreactive moiety, e.g., rhodamine, phenol red, orange azo dye, paparazzi red, non-sulfonated cyanine, sulfonated cyanine, chemiluminescent fluoride sensors (1,2-dioxetane derivatives), and D2A dyes (NIR fluorescent dyes). Alternatively, the photoreactive moiety may be selected from compounds having functional groups and structures similar to the following:

[0894]

[0895] Here:

[0896] R 100 is H or C1-C6-alkyl;

[0897] R 101 is H or SO3H; R 102 is a C1-C6-alkyl or -(CH2) z It is COOH;

[0898] z is an integer from 3 to 8;

[0899] R 103 and R 104 Each is independently H or C1-C6 alkyl; and

[0900] R 105 and R 106 Each is independently hydrogen, COOH, or SO3H.

[0901] Additional photoreaction moiety is disclosed in the relevant technology. for example , Org . Lett. 2014, 16, 1680-1683; J. Am. Chem . Soc . 2011, 133, 10960-10965; Dye Lasers , 3rd Ed. (Springer-Verlag, Berlin, 1990); J. Am. Chem . Soc . See 2012, 134, 20412-20420).

[0902] Treatment methods

[0903] Targeted therapy

[0904] The targeting moiety of the conjugate can be recognized by cells, thereby providing so-called targeted therapy.

[0905] In some embodiments, the conjugate comprises an activator for use in targeted therapy to treat autoimmune diseases. In some such embodiments, the activator is selected from: cyclosporine, cyclosporine A, mycophenylate mofetil, sirolimus, tacrolimus, enenercept, prednisone, azathioprine, methotrexate cyclophosphamide, aminocaproic acid, chloroquine, hydroxychloroquine, hydrocortisone, dexamethasone, chlorrambucil, DHEA, danazol, bromocriptine, meloxicam, infliximab, etc. .

[0906] In some embodiments, the compound includes activator Q for use in targeted therapy to treat infectious diseases. In some such embodiments, Q is selected from the following: beta-lactam series (penicillin G, penicillin V, cloxacillin, dicloxacillin, methicillin, naphsyllin, oxacillin, ampicillin, amoxicillin, becampicillin, azlosillin, carbenicillin, mezlosillin, piperacillin, ticacillin), aminoglycoside series (amikacin, gentamicin, kanamycin, neomycin, netilmycin, streptomycin, tobramycin), macrolide series (azithromycin, clarithromycin, erythromycin, lincomycin, clindamycin), tetracycline series (demeclocycline, doxycycline, minocycline, tetracycline), quinolone series (cinoxacin, nalidixic acid), fluoroquinolone series (ciprofloxacin, Enoxacin, Grepafloxacin, Levofloxacin, Lomefloxacin, Norfloxacin, Ofloxacin, Sparfloxacin, Trovafloxacin), Polypeptide series (Bacitracin, Colistin, Polymyxin B), Sulfonamide series (Sulfisoxazole, Sulfamethoxazole, Sulfadiazine, Sulfamethizol, Sulfacetamide), Other antibiotics (Trimethoprim, Silpamethazole, Chloramphenicol, Vancomycin, Metronidazole, Quinupristin, Talfopristin, Rifampicin, Spectinomycin, Nitrofurantoin), General antivirals (Idoxuradin, Vidarabine, Acyclovir, Famcicyclovir, Pencicyclovir, Valacyclovir, Gangcicyclovir, Foscarnet, Ribavirin, Amantadine, Rimantadine, Cydofovir, antisense oligonucleotides, immunoglobulins, interpheon), HIV infection treatment drugs (tenofovir, emtricitabine, zidovudine, didanosine, zalcitabine, stavudine, lamivudine, nevirapine, delaviridine, saquinavir, ritonavir, indinavir, nelfinavir), etc.

[0907] In some embodiments, the compounds and conjugates disclosed herein comprise an activator Q for use in a method of delivering an activator to cells to treat a tumor, wherein the targeting moiety is selected to bind to target cells (i.e., cancer cells). In particular, the compounds, conjugates, and compositions inhibit abnormal cell growth or mammals ( for example It may be useful for treating proliferative disorders in humans, for example, the target cell is a cancer cell, and the targeting moiety is selected to bind to a molecule that is associated with cancer cells (and is not associated with healthy cells, or is at least preferentially associated with tumor cells rather than healthy cells).

[0908] In some such embodiments, the activator is selected from the following: cytotoxic or immunomodulatory agents, anticancer agents, anti-tubulin agents, cytotoxic drugs, etc. Preferably, the cytotoxic or immunomodulatory agent is an anti-tubulin agent, auristatin, DNA narrow groove binder, DNA transcription inhibitor, alkylating agent, anthracycline, antibiotic, antifoliant, an antimetabolite, calmodulin inhibitor, chemotherapy sensitizer, duocarmycin, etoposide, fluorinated pyrimidine, ion-permeable carrier, lexitropsin, metancinoid, nitrosourea, platinol, pore-forming compound, purine antimetabolite, puromycin, radiosensitizer, rapamycin, steroid, taxane, topoisomerase inhibitor, vinca alkaloid, etc.Includes; anticancer drugs include methotrexate, taxol, L-asparaginase, mercaptopurine, thioguanine, hydroxyurea, cytarabine, cyclophosphamide, ifosfamide, nitrosourea, cisplatin, carboplatin, mitomycin, dacarbazine, procarbazine, topotecan, nitrogen mustard, cyclophosphamide, etoposide, 5-fluorouracil, BCNU, irinotecan, camptothecin, bleomycin, doxorubicin, idarubicin, daunorubicin, dactinomycin, plicamycin, mitoxantrone, asparaginase, vinblastine, vincristine, vinorelbine, paclitaxel, docetaxel, etc. Includes; anti-tubulin agents are taxanes ( for example , paclitaxel, docetaxel), T67, vinca alkaloid ( for example , vincristine, vinblastine, vindecin, vinorelbine), bacatin derivatives, taxane derivatives, epitylon ( for example , Epotillon A, Epotillon B), Nocodazole, Colchicine, Colsimide, Estramustine, Cryptophysin, Semadotine, Meitansinoid, Combrestatin, Discordermolide, Eleutherobin, Auristatin derivatives (AFP, MMAF, MMAE), etc.Includes; cytotoxic drugs include androgens, antramycin (AMC), asparaginase, 5-azacitidine, azathioprine, bleomycin, busulfan, butionine sulfoximine, calichiamycin, calichiamycin derivatives, camptothecin, carboplatin, camustine (BSNU), CC-1065, chlorambucin, cisplatin, colchicine, cyclophosphamide, cytarabine, cytidine arabinoside, psychocalacin B, dacarbazine, dactinomycin (actinomycin), daunorubicin, decarbazine, DM1, DM4, docetaxel, doxorubicin, etoposide, estrogens, 5-fluordeoxyuridine, 5-fluorouracil, gemcitabine, gramycidin D, hydroxyurea, idarubicin, ifosfamide, Irinotecan, lomustine (CCNU), metansine, mechlorethamine, melphalan, 6-merceptorpurine, methotrexate, mitramycin, mitomycin C, mitoxantrone, nitroimidazole, paclitaxel, palitoxin, plicamycin, procarbizine, rhizoxin, streptozotocin, tenofoside, 6-thioguanine, thiotepa, topotecan, vinblastine, vincristine, vinorelbine, VP-16, VM-26; DNA narrow groove binder ( for example , endiine, lexitropsin, CBI compounds), duocarmycin, taxane ( for example , paclitaxel, docetaxel), puromycin, vinca alkaloid, CC-1065, SN-38, topotecan, morpholino-doxorubicin, rhizoxin, cyanomopolino-doxorubicin, echinomycin, combretastatin, netropsin, epotillon A, epotillon B, estramustine, cryptophysin, semadotin, meitansinoid, discodermolide, eleuterobin, mitoxantrone, etc. Includes

[0909] Cell proliferation and apoptosis

[0910] The compounds and conjugates disclosed in this specification may be used in a method for inducing apoptosis in cells.

[0911] Dysregulated apoptosis has been implicated in various diseases, including, for example: autoimmune disorders ( for example , systemic lupus erythematosus, rheumatoid arthritis, graft-versus-host disease, myasthenia gravis, or Sjögren's syndrome), chronic inflammatory conditions ( for example , psoriasis, asthma, or Crohn's disease), hyperproliferative disorders ( for example , breast cancer, lung cancer), viral infection ( for example , herpes, papilloma, or HIV), and other conditions, such as osteoarthritis and atherosclerosis. The compounds, conjugates, and compositions described herein may be used to treat or improve any of these diseases. Such treatment generally involves administering an amount of the compounds, conjugates, or compositions described herein sufficient to provide therapeutic benefit to a subject suffering from the disease. The identity of the antibodies of the administered compounds, conjugates, or compositions will depend on the disease being treated—and thus said antibodies must bind to cell-surface antigens expressed on cell types where inhibition is beneficial. The therapeutic benefit achieved will also depend on the specific disease being treated. In certain cases, the compounds and compositions disclosed herein may treat or improve the disease itself or the symptoms of the disease when administered as monotherapy. In other instances, the compounds and compositions disclosed herein may be part of an overall therapeutic regimen comprising an inhibitor or other agents that treat or improve the disease being treated or the symptoms of the disease, together with the compounds and compositions disclosed herein. Agents useful for treating or improving specific diseases that may be administered in conjunction with or together with the compounds and compositions disclosed herein will be apparent to those skilled in the art.

[0912] While absolute cure is always desirable in any treatment regimen, achieving a cure is not required to provide therapeutic benefit. Therapeutic benefit may include halting or slowing disease progression, regressing the disease without a cure, and / or improving or slowing the progression of disease symptoms. Long-term survival compared to the statistical mean and / or improved quality of life may also be considered a therapeutic benefit.

[0913] Cancer is a specific class of diseases that involve abnormally regulated apoptosis and constitute a significant global health burden. In certain embodiments, the compounds and compositions disclosed herein may be used to treat cancer. Cancer may be, for example, a solid tumor or a hematological tumor. Cancers that may be treated with the compounds and compositions disclosed herein include, but are not limited to, bladder cancer, brain cancer, breast cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myeloid leukemia, multiple myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, multiple myeloma, prostate cancer, small cell lung cancer, and splenic cancer. The compounds and compositions disclosed herein may be particularly beneficial in the treatment of cancer because antibodies can be used to specifically target tumor cells, thereby avoiding or improving undesirable side effects and / or toxicity that may be associated with the systemic administration of potentially unconjugated inhibitors. One embodiment relates to a method for treating a disease involving abnormally controlled apoptosis, said method comprising administering an amount of the compounds and compositions disclosed herein effective for providing therapeutic benefit to a subject having a disease involving abnormally controlled apoptosis, wherein the ligands of the compounds and compositions disclosed herein bind to cell surface receptors on cells having abnormally controlled apoptosis. One embodiment relates to a method for treating cancer comprising the step of administering the compounds and compositions disclosed herein to a subject having cancer, said method, wherein the ligands may bind to cell surface receptors or tumor-associated antigens expressed on the surface of cancer cells in an amount effective for providing therapeutic benefit.

[0914] In the context of tumorigenic cancer, therapeutic benefits, in addition to the effects discussed above, may also specifically include halting or slowing the progression of tumor growth, regression of tumor growth, eradication of one or more tumors, and / or increased patient survival compared to the statistical mean for the type and stage of the cancer being treated. In one embodiment, the cancer being treated is tumorigenic cancer.

[0915] The compounds and conjugates disclosed herein may be administered as monotherapy to provide therapeutic benefits, or may be administered adjunctively to or together with other chemotherapy agents and / or radiotherapy. Chemotherapy agents to which the compounds and compositions disclosed herein may be used as adjunctive therapy may be targeted (e.g., ADCs, protein kinase inhibitors, etc. ) or can be non-targeted (e.g., non-specific cytotoxic drugs such as radionucleotides, alkylating agents, and intermediaries). Non-targeted chemotherapeutic agents to which the compounds and compositions disclosed herein may be administered together comprise, but are not limited to: methotrexate, taxol, L-asparaginase, mercaptopurine, thioguanine, hydroxyurea, cytarabine, cyclophosphamide, ifosfamide, nitrosourea, cisplatin, carboplatin, mitomycin, dacarbazine, procarbizine, topotecan, nitrogen mustard, cyclosan, etoposide, 5-fluorouracil, BCNU, irinotecan, camptothecin, bleomycin, doxorubicin, idarubicin, daunorubicin, dactinomycin, plicamycin, mitoxantrone, asparaginase, vinblastine, vincristine, vinorelbine, paclitaxel, calichiamycin, and docetaxel.

[0916] Compounds and conjugates disclosed herein, which may not be effective as monotherapy for treating cancer, may be administered in addition to or together with other chemotherapy agents or radiotherapy to provide therapeutic benefits. One embodiment relates to a method in which a compound or composition disclosed herein is administered in an amount effective for sensitizing tumor cells to standard chemotherapy and / or radiotherapy. Accordingly, in the context of cancer treatment, "therapeutic benefits" include the administration of the compounds and compositions disclosed herein in addition to or together with chemotherapy agents and / or radiotherapy as a means of sensitizing tumors to either chemotherapy and / or radiotherapy, in patients who have not yet started such therapy or have not yet shown signs of resistance, or in patients who have begun to show signs of resistance.

[0917] Pharmaceutical composition and administration thereof

[0918] The compounds and conjugates disclosed herein may be used to treat an individual in need. In certain embodiments, the individual is a mammal, e.g., a human, or a non-human mammal. When administered to an animal, e.g., a human, the composition or compound is preferably administered as a pharmaceutical composition comprising, for example, the disclosed compound and a pharmaceutically acceptable carrier.

[0919] Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or physiologically buffered saline or other solvents or vehicles such as glycol, glycerol, oils such as olive oil, or injectable organic esters. In a preferred embodiment, where such a pharmaceutical composition is for human administration, particularly for an invasive route of administration (i.e., a route that avoids transport or diffusion through the epithelial barrier, e.g., injection or implantation), the aqueous solution is pyrogen-free or substantially pyrogen-free. Excipients may be selected, for example, to enable delayed release of the formulation or to selectively target one or more cells, tissues, or organs. The pharmaceutical composition may be in dosing unit forms such as tablets, capsules (including sprinkle capsules and gelatin capsules), granules, hydrophilic substances for reconstitution, powders, solutions, syrups, suppositories, injections, etc. The composition may also be present in a transdermal delivery system, for example, a skin patch. The composition may also be present in a solution suitable for topical administration, such as an ointment or cream.

[0920] A pharmaceutically acceptable carrier may contain, for example, a physiologically acceptable agent that acts to increase the absorption of a compound, such as the compound of the present invention, or to stabilize or increase its solubility. Such physiologically acceptable agents include, for example, carbohydrates, such as glucose, sucrose, or dextran; antioxidants, such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins; or other stabilizers or excipients. The selection of a pharmaceutically acceptable carrier containing a physiologically acceptable agent depends, for example, on the route of administration of the composition. The preparation of the pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) may also be a liposome or other polymer matrix, for example, a compound of the present invention, which may be incorporated therein. For example, liposomes containing phospholipids or other lipids are non-toxic, physiologically acceptable, and metabolizable carriers that are relatively simple to prepare and administer.

[0921] The phrase “pharmaceuticalally acceptable” is used herein to refer to a compound, substance, composition, and / or dosage form suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reaction, or other problems or complications, in proportion to a reasonable benefit / harm ratio, within the scope of sound medical judgment.

[0922] As used herein, the phrase “pharmaceuticalally acceptable carrier” means a pharmaceutically acceptable substance, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be “acceptable” in the sense that it is compatible with other components of the formulation and is not harmful to the patient. Some examples of substances that may serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycol, e.g., propylene glycol; (11) polyol, e.g., glycerin, sorbitol, mannitol, and polyethylene glycol; (12) ester, e.g., ethyl oleate and ethyl laurate; (13) agar; (14) buffer, e.g., magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) non-pyrogenous water; (17) isotonic saline solution; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solution; and (21) other non-toxic compatible substances used in pharmaceutical formulations.

[0923] The pharmaceutical composition (formulation) may be administered to a subject by any number of routes of administration, for example, including: orally (e.g., as a drencher, such as in aqueous or non-aqueous solution or suspension, tablet, capsule (including sprinkle capsules and gelatin capsules), bolus, powder, granule, or paste for application to the tongue); absorption via oral mucosa (e.g., sublingually); anally, rectally, or vaginally (e.g., as a pessary, cream, or foam); parenterally (e.g., as a sterile solution or suspension, including intramuscularly, intravenously, subcutaneously, or intrathecally); nasally; intraperitoneally; subcutaneously; transdermally (e.g., as a patch applied to the skin); and topically (e.g., as a cream, ointment, or spray applied to the skin, or as eye drops). The compound may also be formulated for inhalation. In certain embodiments, the compound may be simply dissolved or suspended in sterile water. Details of appropriate routes of administration and compositions suitable for the same may be found in: for example, U.S. Patent Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896, as well as the patents cited herein.

[0924] The formulation can be conveniently presented in a unit dosage form and can be manufactured by any method well known in the field of compounding. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending on the host being treated and the specific method of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound producing a therapeutic effect. Generally, out of 100 percent, this amount will be in the range of about 1 percent to about 99 percent, preferably about 5 percent to about 70 percent, most preferably about 10 percent to about 30 percent of the active ingredient.

[0925] A method for preparing these formulations or compositions comprises the step of associating an active compound, such as a compound of the present invention, with a carrier and, optionally, one or more auxiliary components. Generally, a formulation is prepared by associating the compound of the present invention uniformly and closely with a liquid carrier, or a finely ground solid carrier, or both, and then, if necessary, by shaping the product.

[0926] The formulation of the present invention suitable for oral administration is a compound of the present invention as an active ingredient. predetermined The compounds, conjugates, or compositions thereof may also be administered as boluses, soft tablets, or pastes, each containing an amount thereof.

[0927] To manufacture solid dosage forms for oral administration (capsules (including sprinkle capsules and gelatin capsules), tablets, pills, coated tablets, powders, granules, and other similar forms), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or disodium calcium phosphate, and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia; (3) humectants, such as glycerol; (4) disintegrants, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retardants, such as paraffin; and (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, e.g., cetyl alcohol and glycerol monostearate; (8) absorbents, e.g., kaolin and bentonite clay; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof; (10) complexing agents, e.g., modified and unmodified cyclodextrin; and (11) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical composition may also include a buffer. A similar type of solid composition may also be used as a filler in soft and hard-filled gelatin capsules using excipients such as lactose or lactose, as well as high molecular weight polyethylene glycol and other of the same.

[0928] Tablets may be made by compression or molding, optionally with one or more auxiliary components. Compressed tablets may be manufactured using a binder (e.g., gelatin or hydroxypropylmethylcellulose), a lubricant, an inert diluent, a preservative, a disintegrant (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), a surfactant, or a dispersant. Molded tablets may be made by molding from a suitable mechanical mixture of powdered compounds moistened with an inert liquid diluent.

[0929] Tablets and other solid dosage forms of the pharmaceutical composition, such as coated tablets, capsules (including sprinkle capsules and gelatin capsules), pills, and granules, may optionally be obtained or manufactured with coatings and shells, such as intestinal coatings and other coatings well known in pharmaceutical formulation technology. They may also be formulated to provide slow or controlled release of the active ingredient within them by using hydroxypropylmethylcellulose, for example, in variable proportions to provide a desired release profile, different polymer matrix, liposomes, and / or microspheres. They may be sterilized, for example, by filtration through a bacteria-fixing filter, or by incorporating a sterile agent in the form of a sterile solid composition that can be dissolved in sterile water or some other sterile injectable medium immediately before use. These compositions may also optionally contain an opacifying agent and may be compositions that release the active ingredient(s) alone, or optionally, preferentially in a delayed manner in a specific part of the gastrointestinal tract. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient may also be in a micro-encapsulated form with one or more of the excipients described above, if appropriate.

[0930] Liquid dosage forms useful for oral administration include pharmaceutically acceptable emulsions, hydrophilic substances for reconstitution, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, the liquid dosage form may contain the following: inert diluents commonly used in the art, e.g., water or other solvents, cyclodextrins and derivatives thereof, solubilizers and emulsifiers, e.g., ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (particularly cottonseed, peanut, corn, bacterium, olive, castor, and sesame oils), glycerol, fatty acid esters of tetrahydrofuryl alcohol, polyethylene glycol, and sorbitan, and mixtures thereof.

[0931] In addition to inert diluents, the oral composition may also include adjuvants such as humectants, emulsifiers and suspending agents, sweeteners, flavoring agents, coloring agents, fragrances and preservatives.

[0932] In addition to the active compound, the suspension may contain a suspending agent, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth, and mixtures thereof.

[0933] A formulation of a pharmaceutical composition for rectal, vaginal, or urethral administration may be presented as a suppository, which may be prepared by mixing one or more active compounds with one or more suitable non-irritating excipients or carriers, such as cocoa butter, polyethylene glycol, suppository wax, or salicylate, which is solid at room temperature but liquid at body temperature, and thus will melt in the rectal or vaginal cavity and release the active compound.

[0934] A formulation of a pharmaceutical composition for oral administration may be presented as a mouthwash, an oral spray, or an oral ointment.

[0935] Alternatively or additionally, the composition may be formulated for delivery via a catheter, stent, wire, or other lumenical device. Delivery via such devices may be particularly useful for delivery to the bladder, urethra, ureter, rectum, or intestine.

[0936] Formulations suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams, or spray formulations containing such carriers as known in the art, where appropriate.

[0937] Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active compound may be mixed with a pharmaceutically acceptable carrier under sterile conditions and with any preservatives, buffers, or propellants that may be required.

[0938] Ointments, pastes, creams, and gels may contain, in addition to the active compound, excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, and zinc oxide, or mixtures thereof.

[0939] The powder and spray may contain, in addition to the active compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures of these materials. The spray may additionally contain conventional propellants such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

[0940] A transdermal patch has the added advantage of providing controlled delivery of the compound of the present invention to the body. Such a dosage form can be made by dissolving or dispersing the active compound in a suitable medium. An absorption enhancer can also be used to increase the flow of the compound through the skin. The rate of such flow can be controlled by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0941] Ophthalmic formulations, eye ointments, powders, solutions, and other like products are also considered to be within the scope of the present invention. Exemplary ophthalmic formulations are described below: U.S. Publication Nos. 2005 / 0080056, 2005 / 0059744, 2005 / 0031697, and 2005 / 004074; and U.S. Patent No. 6,583,124 (the contents of which are incorporated herein by reference). If desired, liquid ophthalmic formulations have properties similar to or compatible with lacrimal fluids, aqueous humor, or vitreous fluids.

[0942] As used herein, the phrases "parenteral administration" and "parenterally administered" " Modes of administration other than intestinal and local administration, generally meaning by injection, and, without limitation, include: intravenous, intramuscular, intra-arterial, intra-spinal, intraorbital, intracardiac, intradermal, intraperitoneal, transtubercular, subcutaneous, subcutaneous, intra-articular, subcapsular, subarachnoid, intraspinal, and intrasternal injections and infusions.

[0943] A pharmaceutical composition suitable for parenteral administration comprises one or more active compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders that may be reconstituted immediately before use into sterile injectable solutions or dispersions containing antioxidants, buffers, bacteriostatic agents, and solutes that make the formulation isotonic with the blood of the intended receiver or suspension or thickener.

[0944] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical composition of the present invention include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, and other similar ones), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Suitable fluidity can be maintained, for example, by the use of a coating material, such as lecithin, by maintaining the particle size required in the case of a dispersion, and by the use of a surfactant.

[0945] These compositions may also contain adjuvants, such as preservatives, wetting agents, emulsifiers, and dispersants. Prevention of microbial action can be ensured by the encapsulation of various antimicrobial and antifungal agents, such as parabens, chlorobutanol, phenol sorbic acid, and other similar substances. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and other similar substances, in the composition. Additionally, long-term absorption in the injectable pharmaceutical form may be achieved by the encapsulation of absorption-delaying agents, such as aluminum monostearate and gelatin.

[0946] In some cases, to prolong the effect of the drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of a crystalline or amorphous material with poor water solubility. The absorption rate of the drug then depends, in turn, on its dissolution rate, which can depend on crystal size and crystal form. Alternatively, delayed absorption of parenterally administered drug forms is achieved by dissolving or suspending the drug in an oil vehicle.

[0947] Injectable depot formulations are created by forming a microencapsulated matrix of the compound in a biodegradable polymer, such as polylactide-polyglycolide. The rate of drug release can be controlled depending on the drug-to-polymer ratio and the properties of the specific polymer used. Other examples of biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by drug capture in liposomes or microemulsions compatible with body tissues.

[0948] For use in the method of the present invention, the active compound may be provided in combination with a pharmaceutically acceptable carrier, for example, as a pharmaceutical composition containing 0.1 to about 99.5% (more preferably about 0.5 to about 90.0%) of the active ingredient, or as itself.

[0949] In some embodiments of the present invention, the compound of the present invention is administered in combination with one or more additional compounds / preparations.

[0950] In certain such embodiments, co-administration is simultaneous. In certain such embodiments, the compound of the present invention is co-formulated with one or more additional compounds. In certain other such embodiments, the compound of the present invention is administered separately but simultaneously with one or more additional compounds. In certain such embodiments, co-administration is sequential, with the administration of the compound of the present invention before or after the administration of one or more additional compounds by minutes or hours.

[0951] The method of introducing the compound of the present invention may also be provided by a rechargeable or biodegradable device. Various sustained-release polymer devices have been developed and have recently been tested in vivo for the controlled delivery of drugs, including proteinaceous biopharmaceuticals. Various biocompatible polymers (including hydrogels), including both biodegradable and non-degradable polymers, may be used to form implants for sustained release of the compound at specific target sites.

[0952] The actual dosage level of the active ingredient in a pharmaceutical composition can be varied to obtain an amount of the active ingredient effective in achieving a desired therapeutic response for a specific patient, composition, and mode of administration without toxicity to the patient.

[0953] The selected dosage level will depend on various factors including: the activity of the specific compound, conjugate or combination of compound and / or conjugate, or its ester, salt, or amide; the route of administration of the specific compound(s) being used, the time of administration, the elimination rate, the duration of treatment; other drugs, compounds, and / or substances used in combination with the specific compound(s) being used; the age, sex, weight, health status, general health and prior medical history of the patient being treated; and similar factors well known in the medical field.

[0954] A physician or veterinarian with ordinary skills in the art can easily determine and prescribe the therapeutic effective dose of the required pharmaceutical composition. For example, a physician or veterinarian may start a dose of the pharmaceutical composition or compound at a lower level than required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved. "Therapeutic effective dose" refers to a concentration of the compound sufficient to induce the desired therapeutic effect. It is generally understood that the effective dose of the compound will vary depending on the subject's weight, sex, age, and medical history. Other factors affecting the effective dose may include, but are not limited to, the following: the severity of the patient's condition, the disorder being treated, the stability of the compound, and, if desired, another type of therapeutic agent being administered with the compound of the present invention. A larger total dose may be delivered by multiple administrations of the formulation. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison's Principles of Internal Medicine 13 ed., 1814-1882, incorporated herein by reference).

[0955] Generally, a suitable daily dose of the active compound used in the composition and method of the present invention will be the amount of the compound that is the lowest effective dose for producing a therapeutic effect. Such an effective dose will generally depend on the factors described above.

[0956] If desired, an effective daily dose of the active compound or conjugate may be administered, optionally, as 1, 2, 3, 4, 5, 6, or greater sub-dose, administered separately at appropriate intervals throughout the day in the form of unit doses. In a specific embodiment of the invention, the active compound may be administered two or three times daily. In a preferred embodiment, the active compound will be administered once daily.

[0957] Patients receiving such treatment are primates, particularly humans, and other mammals such as horses, cattle, pigs, and sheep; and any animals in need, generally including poultry and pets.

[0958] In certain embodiments, the compounds or conjugates disclosed herein may be used alone or administered co-administered with another type of therapeutic agent. As used herein, the phrase “co-administered” refers to any form of administration of two or more different therapeutic compounds or conjugates such that a second compound or conjugate is administered while the previously administered therapeutic compound or conjugate is still effective in the body (e.g., two compounds or conjugates are simultaneously effective in the patient, which may include a synergistic effect of the two compounds or conjugates). For example, different therapeutic compounds or conjugates may be administered simultaneously or sequentially within the same formulation or within separate formulations. In certain embodiments, different therapeutic compounds or conjugates may be administered within 1 hour, 12 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, weeks, or longer than that. Thus, an individual receiving such treatment may benefit from the combined effect of different therapeutic compounds or conjugates.

[0959] The present invention includes the use of pharmaceutically acceptable salts of the compounds or conjugates disclosed herein. In certain embodiments, the salts considered in the present invention include, but are not limited to, the following: alkyl, dialkyl, trialkyl, or tetraalkyl ammonium salts. In certain embodiments, the salts considered of the present invention comprise, but are not limited to: L-arginine, benetamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucarmine, hydravamin, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain embodiments, the salts considered of the present invention comprise, but are not limited to: Na, Ca, K, Mg, Zn, or other metal salts.

[0960] Pharmaceutically acceptable acid addition salts may also exist as various solvates with, for example, water, methanol, ethanol, dimethylformamide, and other similar substances. Mixtures of such solvates may also be prepared. The source of such solvates is derived from the solvent of crystallization and may be inherent to the solvent of preparation or crystallization, or may be incidental to such solvent.

[0961] Wetting agents, emulsifiers, and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweeteners, flavoring agents and fragrances, preservatives, and antioxidants may also be present in the composition.

[0962] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and other similar types; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol and other similar types; and (3) metal-chelating agents, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid and other similar types.

[0963] The present invention will be more easily understood by reference to the following examples, which are included for the purpose of illustrative examples of specific aspects and embodiments of the invention and are not intended to limit the invention.

[0964] illustration

[0965] Synthesis protocol

[0966] abbreviation

[0967] AcO : Acetyl

[0968] AcOH: Acetic acid

[0969] EA: Ethyl acetate

[0970] DCM: Dichloromethane

[0971] m-CPBA: Meta-chloroperoxybenzoic acid

[0972] TBDMSOTf : tert-butyldimethylsilyl trilate

[0973] TBDMS: tert-butyldimethylsilyl

[0974] DMF: Dimethylformamide

[0975] EDCI : 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide

[0976] HOBt: 1-Hydroxybenzotriazole hydrate

[0977] ACN: Acetonitrile

[0978] TBDMS-Cl : tert-butyldimethylsilyl chloride

[0979] DBU : 1,8-Diazabicyclo[5.4.0]Undek-7-en

[0980] THF: Tetrahydrofuran

[0981] DCC : N,N'-Dicyclohexylcarbodiimide

[0982] DMAP: 4-Dimethylaminopyridine

[0983] NHS: N-hydroxysuccinimide

[0984] DIPEA: Diisopropylethylamine

[0985] TEA: Triethylamine

[0986] DEAD: Diethyl azodicarboxylate

[0987] Boc: tert-butyloxycarbonyl

[0988] LAH: Lithium Aluminum Hydrate

[0989] CDI: 1,1'-Carbonyldiimidazole

[0990] BEMP: 2-tert-butylimino-2-diethylamino-1,3-dimethylperhydro-1,3,2-diazaphosphorin

[0991] TPSCl : Triphenylchlorosilane

[0992] tfa: trifluoroacetyl

[0993] PyBop : Benzotriazole-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate

[0994] HBTU : N,N,N',N'-tetramethyl-O-(1H-benzotriazole-1-yl)uronium hexafluorophosphate

[0995] TFA: Trifluoroacetic acid

[0996] DIC : N,N'-Diisopropylcarbodiimide

[0997] DMPA: 2,2-Dimethoxy-2-phenylacetophenone

[0998] TBAF: Tetra-n-butylammonium fluoride

[0999] AgOTf: Silver trifluoromethanesulfonate

[1000] (BimC4A)3: Tripotassium 5,5',5"-[2,2',2"-Nitrilotris(methylene)tris(1 H -Benzimidazole-2,1-diyl)]Tripentanoate hydrate

[1001] [Example 1] Int-TG manufacturing

[1002]

[1003] β-D-galactose pentaacetate (Alfa, CAS 4163-60-4, 5.0 g, 12.81 mmol) was dissolved in 33% HBr in AcOH (20 mL) under an N2 atmosphere at 0°C. The mixture was heated to room temperature. After stirring at room temperature for 4 hours, the mixture was concentrated under reduced pressure, and then EA (1000 mL) and saturated sodium bicarbonate (1000 mL) were added. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to yield the compound Int-TG (5.2 g, 99%).

[1004] 1 ¹H NMR (400 MHz, CDCl₃) δ 6.70 (d, J = 4.0 Hz, 1H), 5.52 (d, J = 2.4 Hz, 1H), 5.41 (dd, J = 7.6, 2.8 Hz, 1H), 5.05 (dd, J = 6.4, 4.0 Hz, 1H), 4.49 (t, J= 6.4 Hz, 1H), 4.22-4.09 (m, 2H), 2.16 - 2.01 (m, 12H).

[1005] [Example 2] Preparation of Compound FA-Int

[1006]

[1007] Compound FA-Int was obtained by a method similar to the method described in U.S. Patent Application Publication No. 2007 / 0276018, the entirety of which is incorporated herein by reference.

[1008] [Example 3] Preparation of Compound IntC-L-1

[1009]

[1010] Preparation of compound IntCl-L-1a

[1011] Boc2O (14.7 g, 67.47 mmol) in DCM (200 mL) was added to a solution of 2,2-(ethylenedioxy)bis(ethylamine) (50 g, 337.4 mmol) in DCM (300 mL) under an N2 atmosphere. The mixture was stirred overnight at room temperature and quenched with H2O (500 mL) and brine (150 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. After concentration, compound IntCl-L-1a was used directly without purification in the following reaction (13.01 78%).

[1012] 1 ¹H NMR (400 MHz, CDCl₃) δ 5.20 (s, 1H), 3.62-3.62 (m, 4H), 3.55-3.51 (m, 4H), 3.35-3.25 (m, 2H), 2.90-2.87 (m, 2H), 1.45 (s, 9H).

[1013] Preparation of compound IntCl-L-1b

[1014] PyBOP (15.72 g, 30.20 mmol) and DIPEA (10.52 mL, 60.39 g, mmol) were added to a solution of compounds IntCl-L-1a (6 g, 24.16 mmol) and zL-Glu-OMe (5.94 g, 20.13 mmol) in DMF (30 mL) under an N2 atmosphere at 0°C. The mixture was stirred at room temperature for 2 hours. EA (20 mL x 6), H2O (20 mL), and brine (200 mL) were added to the mixture. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to yield compounds IntCl-L-1b (10.6 g, quant.).

[1015] 1 ¹H NMR (400 MHz, CDCl₃) δ 7.40-7.28 (m, 5H), 6.32 (s, 1H), 5.80 (s, 1H), 5.11 (s, 2H), 5.02 (s, 1H), 4.36 (s, 1H), 3.74 (s, 3H), 3.60 (s, 4H), 3.54 (s, 4H), 3.44-3.43 (m, 2H), 3.38-3.21 (m, 2H), 2.30-2.20 (m, 3H), 2.04-2.00 (m, 1H), 1.76 (s, 1H), 1.44 (s, 9H). EI-MS m / z: 526 (M + ).

[1016] Preparation of compound IntCl-L-1c

[1017] Pd / C (900 mg) was added to a solution of compound IntCl-L-1b (3 g, 5.71 mmol) in MeOH (25 mL) at room temperature under H2. The mixture was stirred for 3 hours, filtered through Celite®, and then concentrated under reduced pressure. Compound IntCl-L-1c was used directly in the next step without further purification (2.23 g, crude material).

[1018] 1¹H NMR (400 MHz, CDCl₃) δ 6.56 (s, 1H), 5.20 (s, 1H), 3.73 (s, 3H), 3.61 (s, 4H), 3.57-3.55 (m, 4H), 3.53-3.50 (m, 1H), 3.48-3.44 (m, 4H), 2.40-2.32 (m, 2H), 2.18-2.10 (m, 1H), 1.88-1.81 (m, 1H), 1.44 (s, 9H). EI-MS m / z: 392 (M + ).

[1019] Preparation of compound IntCl-L-1d

[1020] Compound IntCl-L-1c (2.23 g, 5.71 mmol) and compound FA-Int (2.12 g, 5.19 mmol) in DMF (15 mL) were added to a solution of HBTU (2.36 g, 6.23 mmol) and DIPEA (1.36 mL, 7.78 mmol) under an N2 atmosphere at 0°C. The mixture was stirred at room temperature for 2.5 hours, and EA (100 mL × 7) and H2O (100 mL) were added. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to yield compound IntCl-L-1d (4.06 g, quant.).

[1021] 1 ¹H NMR (400Hz, DMSO-d6) δ 8.89 (d, J =7.6 Hz, 1H), 8.63 (s, 1H), 7.90 (d, J =8 Hz, 2H), 7.64 (d, J =8.4 Hz, 2H), 6.76-6.75 (m, 1H), 5.12 (s, 1H), 4.41-4.36 (m, 1H), 3.64 (s, 3H), 3.47 (s, 4H), 3.39-3.35 (m, 4H), 3.20-3.12 (m, 2H), 3.07-3.02 (m, 2H), 2.23 (t,J =7.4 Hz, 2H), 2.09-2.06 (m, 1H), 1.96-1.91 (m, 1H), 1.36 (s, 9H). EI-MS m / z: 782 (M + ).

[1022] Preparation of Compound IntCl-L-1

[1023] TFA (10 mL) was added dropwise at 0°C to a solution of compound IntCl-L-1d (4.68 g, 5.99 mmol) in DCM (50 mL). The reaction was allowed to warm to room temperature and stirred for 3 hours. The mixture was concentrated under reduced pressure and used directly in the next step without further purification (4.08 g, crude material).

[1024] EI-MS m / z: 682 (M + ).

[1025] [Example 4] Preparation of Compound IntC-L

[1026]

[1027] Preparation of Compound K-1

[1028] PyBop (7.89 g, 15.16 mmol) was added at once to L-Lys(Boc)-OMe (3 g, 10.11 mmol) and 4-pentinoic acid (992 mg, 10.11 mmol) in DMF (30 mL), followed by the addition of DIPEA (5.26 mL, 30.32 mmol) under an N2 atmosphere at 0°C. The mixture was stirred overnight at room temperature. EA (80 mL x 4) and saturated citric acid (60 mL) were added to the mixture, and the organic layer was washed with NaHCO3 (120 mL) and brine (100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to yield compound K-1 (3.29 g, 95%).

[1029] EI-MS m / z: 341 (M + ).

[1030] Preparation of Compound K-2

[1031] LiOH·H2O (2.03 g, 48.32 mmol) dissolved in H2O (15 mL) was added to a solution of compound K-1 (3.29 g, 9.66 mmol) in MeOH (15 mL) under an N2 atmosphere at 0℃.

[1032] The mixture was stirred at 0°C for 30 minutes and heated to room temperature for 2 hours. The mixture was acidified with a saturated aqueous citric acid solution, and EA (40 mL x 2) was added to the mixture. The organic layer was washed with H2O (30 mL) and brine (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. Compound K-2 was used directly in the next step without further purification (3.15 g, crude material).

[1033] EI-MS m / z: 327 (M + ).

[1034] Preparation of Compound K

[1035] NHS (1.69 mg, 14.7 mmol) and EDCI (2.93 g, 15.26 mmol) were added to a solution of Compound K-2 (3.69 g, 11.31 mmol) in DMF (20 mL) under an N2 atmosphere. The mixture was stirred and concentrated overnight at room temperature. The remainder, Compound K, was used directly in the next step without further purification (4.79 g, crude material).

[1036] EI-MS m / z: 446 (M + +Na).

[1037] Preparation of Compound IntC-L-2

[1038] DIPEA (5.21 mL, 29.93 mmol) was added to a solution of compound IntC-L-1 (4.08 g, 5.99 mmol) and compound K (4.79 g, 11.31 mmol) in DMF (25 mL) under an N2 atmosphere. The mixture was stirred overnight at room temperature. H2O (70 mL) and brine (60 mL) were added to the mixture and extracted with EA (70 mL x 7). The organic layer was then dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to yield compound IntC-L-2 (1.12 g, 19%).

[1039] EI-MS m / z: 991 (M + ).

[1040] Preparation of Compound IntC-L-3

[1041] LiOH·H2O (356 mg, 8.48 mmol) dissolved in H2O (10 mL) was added to a solution of compound IntA-L-2 (1.12 g, 1.13 mmol) in MeOH (27 mL) under an N2 atmosphere at 0°C. The mixture was stirred at 0°C for 30 minutes and heated to room temperature for 3 hours. The mixture was acidified with 2M HCl and concentrated under reduced pressure. The remaining compound IntC-L-3 was used directly in the next step without further purification (996 mg, crude material).

[1042] EI-MS m / z: 880 (M +).

[1043] Compound IntC-L manufacturing

[1044] TFA (8 mL) was added to a solution of compound IntC-L-3 (996 mg, 1.13 mmol) in DCM (30 mL) under an N2 atmosphere at 0°C. After stirring for 1 hour at 0°C, the mixture was concentrated under reduced pressure. The residue was dissolved in DMSO (5 mL) and purified by Prep-HPLC to produce compound IntC-L (409 mg, 32%).

[1045] EI-MS m / z: 780 (M + ).

[1046] [Example 5] Preparation of Compound MPS-D1

[1047]

[1048] Preparation of compound MPS-D1a

[1049] Piperidine hydrochloride (6.66 g, 54.82 mmol), paraformaldehyde (4.95 g, 164.5 mmol), and conc. HCl (0.6 mL) were added to a solution of 4-acetylbenzo (9 g, 54.82 mmol) in EtOH (50 mL) under an N2 atmosphere at room temperature. The mixture was stirred at 100°C for 16 hours, cooled to room temperature, and acetone (90 mL) was added dropwise to the mixture. The mixture was stirred at 0°C for 1 hour. The solid was filtered and washed with diethyl ether (30 mL x 2) to yield compound MPS-D1a (6.11 g, 38%).

[1050] 1 ¹H NMR (400 MHz, DMSO-d6) δ 8.08 (s, 4H), 5.73 (s, 1H), 3.65 (t, J = 7.2 Hz, 2H), 3.35 (t, J = 7.2 Hz, 2H), 3.31 (m, 6H), 1.74 (s, 4H).

[1051] Preparation of compound MPS-D1b

[1052] 4-methoxybenzenethiol (2.55 g, 20.52 mmol) and piperidine (0.3 mL, 3.08 mmol) were added at room temperature to a solution of MPS-D1a (6.11 g, 20.52 mmol) in EtOH (40 mL) and MeOH (26 mL). The mixture was stirred at 100°C for 16 hours, then cooled to 0°C and stirred for an additional 1 hour. The solid was filtered and washed with ether (30 mL x 2) to yield compound MPS-D1b (5.56 g, 90%).

[1053] 1 ¹H NMR (400 MHz, CDCl₃) δ 8.04-7.99 (m, 4H), 7.27 (d, J = 8.4 Hz, 2H), 7.15 (d, J = 7.6 Hz, 2H), 3.39-3.36 (m, 2H), 3.25-3.21 (m, 2H), 2.27 (s, 3H).

[1054] Preparation of Compound MPS-D1

[1055] Oxone (25.03 g, 40.72 mmol) was added to a solution of MPS-D1b (5.56 g, 18.51 mmol) in MeOH (90 mL) and distilled water (90 mL) under an N2 atmosphere at 0°C. After stirring for 14 hours at room temperature, the mixture was quenched with distilled water (100 mL) and chloroform (150 mL x 3). The organic layer was washed with brine (200 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to yield compound MPS-D1 (5.29 g, 86%).

[1056] 1 ¹H NMR (400 MHz, CDCl₃) δ 8.04-7.99 (m, 4H), 7.81 (d, J = 8.4 Hz, 2H), 7.46 (d, J = 8.4 Hz, 2H), 3.63 (t, J= 7.2 Hz, 2H), 3.41 (t, J = 7.2 Hz, 2H), 2.44 (s, 3H). EI-MS m / z: 333 (M + ).

[1057] [Example 6] Preparation of Compound MPS-D2

[1058]

[1059] Preparation of compound L-1a

[1060] KI (294 mg, 1.77 mmol) and Ag2O (4.92 g, 19.48 mmol) were added to a solution of hexaethylene glycol (5.0 g, 17.71 mmol) in anhydrous DCM (178 mL) under an N2 atmosphere. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was filtered through Celite® and washed with DCM (100 mL). The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography to yield compound L-1a (5.98 g, 73%).

[1061] 1 ¹H NMR (400 MHz, CDCl₃) δ 7.80 (d, J = 8.4 Hz, 2H), 7.35 (d, J = 8.4 Hz, 2H), 4.16 (t, J = 4.8 Hz, 2H), 3.71-3.58 (m, 22H), 2.88 (br, 1H), 2.45 (s, 3H).

[1062] Preparation of compound L-1b

[1063] NaN3 (1.34 g, 20.55 mmol) was added to a solution of compound L-1a (5.98 g, 13.7 mmol) in DMF (30 mL) under an N2 atmosphere. The mixture was stirred at 110°C for 1 hour and concentrated under reduced pressure. The residue was purified by column chromatography to yield compound L-1b (4.1 g, 97%).

[1064] 1 ¹H NMR (400 MHz, CDCl₃) δ 3.72-3.60 (m, 22H), 3.39 (t, J = 4.8 Hz, 2H), 2.78 (br, 1H).

[1065] Preparation of compound L-1c

[1066] Jones reagent solution (5 mL) was slowly added dropwise to a solution of compound L-1b (2 g, 6.51 mmol) in acetone (56 mL) under an N2 atmosphere at -5°C. The mixture was stirred at room temperature for 2 hours, filtered through celite, and the filtrate was concentrated under reduced pressure. The filtrate was diluted with DCM (20 mL × 2) and water (5 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to yield compound L-1c (1.85 g, 89%).

[1067] 1 ¹H NMR (400 MHz, CDCl₃) δ 4.15(s, 2H), 3.76-3.67 (m, 18H), 3.40 (t, J = 4.8 Hz, 2H).

[1068] Preparation of compound L-1d

[1069] t-BuOH (305 μL, 3.11 mmol), DIC (292.5 μL, 1.87 mmol), and DMAP (19 mg, 0.16 mmol) were added to a solution of compound L-1c (500 mg, 1.56 mmol) in DCM (10 mL) under an N2 atmosphere. The mixture was stirred at room temperature for 4 hours and diluted with DCM (30 mL × 2). The organic layer was washed with water (5 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to yield compound L-1d (278.5 mg, 47%).

[1070] 1 ¹H NMR (400 MHz, CDCl₃) δ 4.01 (s, 2H), 3.70-3.66 (m, 18H), 3.38 (t, J = 4.8 Hz, 2H), 1.47 (s, 9H).

[1071] Preparation of compound L-1e

[1072] Pd / C (236 mg, 0.11 mmol) and 4M-HCl (in 1,4-dioxane) were added to a solution of compound L-1d (278 mg, 0.74 mmol) in EtOH (5 mL) under an N2 atmosphere. The mixture was stirred at room temperature for 1 hour. The mixture was filtered through Celite® to remove Pd / C and concentrated to yield compound L-1e (255.3 mg, 89.2%).

[1073] 1 ¹H NMR (400 MHz, DMSO-d6) δ 8.32 (s, 1H), 3.98 (s, 2H), 3.55-3.40 (m, 18H), 3.86 (t, J = 5.6 Hz, 2H), 2.70-2.64 (m, 2H), 1.42 (s, 9H).

[1074] Preparation of Compound MPS-D2

[1075] HBTU (300 mg, 0.79 mmol) and DIPEA (229.3 μL, 1.32 mmol) were added to a solution of compound L-1e (255.3 mg, 0.66 mmol) and compound MPS-D1 (240.6 mg, 0.72 mmol) in DMF (6 mL) under a nitrogen atmosphere. The mixture was stirred at room temperature for 2 hours and diluted with EA (20 mL × 2) and water (5 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to yield compound MPS-D2 (306 mg, 71%).

[1076] 1 ¹H NMR (400 MHz, CDCl₃) δ 7.95 (s, 4H), 7.82 (d, J = 8.0 Hz, 2H), 7.38 (d, J = 8.0 Hz, 2H), 7.33-7.30 (m, 1H), 3.98 (s, 2H), 3.68-3.63 (m, 18H), 3.55-3.53 (m, 2H), 3.49-3.47 (m, 2H), 2.95 (s, 1H), 2.88 (s, 1H), 2.46 (s, 3H) 1.46 (s, 9H). EI-MS m / z: 666(M + +1).

[1077] [Example 7] Preparation of Compound MPS-D4

[1078]

[1079] Preparation of Compound MPS-D3

[1080] TFA (4 mL) was added to a solution of compound MPS-D2 (120 mg, 0.18 mmol) in DCM (8 mL) at 0°C. The reaction was allowed to proceed over 2 hours under an N2 atmosphere and heated to room temperature. After the reaction was complete, the mixture was concentrated three times under reduced pressure using toluene as an auxiliary solvent to remove the TFA. The mixture was then redissolved in DMF, and NHS (31 mg, 0.27 mmol) and EDCI (52 mg, 0.27 mmol) were added. The mixture was stirred overnight at room temperature. After the reaction was complete, compound MPS-D3 was used directly in the next step without further purification (127 mg, crude material).

[1081] EI-MS m / z: 707 (M + ).

[1082] Preparation of Compound MPS-D4

[1083] DIPEA (112 μL, 0.64 mmol) was added to a solution of compound IntC-L (60 mg, 0.08 mmol) and compound MPS-D3 (82 mg, 0.12 mmol) in DMF (6 mL) under an N2 atmosphere. The mixture was stirred for 30 minutes, dissolved in DMSO (3 mL), and purified by HPLC to obtain compound MPS-D4 (77 mg, 73%).

[1084] EI-MS m / z: 1373 (M + ).

[1085] [Example 8] Preparation of Compound MPS-D5

[1086]

[1087] Propargylamine (106 μL, 1.65 mmol) was added to a solution of compound MPS-D1 (500 mg, 1.50 mmol) in DMF (8 mL) under an N2 atmosphere at room temperature. The reaction was cooled to 0°C, and PyBop (1.17 g, 2.26 mmol) and DIPEA (524 μL, 3.01 mmol) were added to the mixture. The mixture was stirred at room temperature for 2 hours and diluted with EA (30 mL × 2) and distilled water (20 mL). The organic layer was extracted, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to yield compound MPS-D5 (510 mg, 92%).

[1088] 1 ¹H NMR (400 MHz, CDCl₃) δ 9.11 (t, J = 5.2 Hz, 1H), 7.98-7.89 (m, 4H), 7.79 (d, J = 8.0 Hz, 2H), 7.43 (d, J = 8.4 Hz, 2H), 4.05-4.03 (m, 2H), 3.60 (t, J= 7.6 Hz, 2H), 3.39 (t, J = 7.2 Hz, 2H), 3.12 (s, 1H), 2.38 (s, 3H).

[1089] [Example 9] Preparation of Compound A-15-1

[1090]

[1091] Preparation of compound A-15-1a

[1092] DCC (1.18 g, 5.71 mmol) and DMAP (92 mg, 0.76 mmol) were added to a solution of z-valine (1.01 g, 3.81 mmol) and N-methylaniline (412 μL, 3.81 mmol) in DCM (15 mL) under an N2 atmosphere at room temperature, followed by stirring at room temperature for 3 hours. The mixture was filtered through Celite® and concentrated under reduced pressure. The residue was purified by column chromatography to yield compound A-15-1a (1.05 g, 78%).

[1093] EI-MS m / z: 584(M + ).

[1094] Preparation of compound A-15-1b

[1095] Compound A-15-1a (1.05 g, 2.96 mmol) was dissolved in MeOH (15 mL) under a nitrogen atmosphere, and Pd / C (378 mg, 0.18 mmol) was added. After stirring under H2 at room temperature for 2 hours, the mixture was filtered through Celite® and washed with MeOH (30 mL). The filtrate was concentrated to yield compound A-15-1b (560 mg, 86.0%).

[1096] 1 ¹H NMR (400 MHz, CDCl₃) δ 7.45-7.41 (m, 2H), 7.38-7.36 (m, 1H), 7.19 (d, J= 7.6 Hz, 1H), 3.32 (s, 3H), 2.88 (d, J = 6.0 Hz, 1H), 2.33 (s, 3H), 1.73 (q, J = 6.8 Hz, 1H), 0.86 (d, J = 6.8 Hz, 3H), 0.80 (d, J = 6.8 Hz, 3H).

[1097] Preparation of Compound A-15-1

[1098] 37% formaldehyde (223 μL, 2.99 mmol) and AcOH (1.14 mL, 19.8 mmol) were added to a solution of compound A-15-1b (220 mg, 0.99 mmol) in DMF (8 mL) under an N2 atmosphere. After stirring for 5 minutes at room temperature, NaCNBH3 (125 mg, 1.98 mmol) was added. The mixture was stirred for 2 hours at room temperature and quenched with saturated NaHCO3 (15 mL x 2). EA (20 mL x 2) and brine (20 mL) added to the mixture were added. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to yield compound A-15-1 (189 mg, 81%).

[1099] EI-MS m / z: 235(M + ).

[1100] [Example 10] Preparation of compound POS-D1

[1101]

[1102] Preparation of compound POS-D1a

[1103] NH2NH2·H2O (88 mL, 1805.4 mmol) was added to a solution of ethyl 4-hydrobenzoate (20 g, 120.35 mmol) in EtOH (60 mL) under an N2 atmosphere. The mixture was stirred overnight under reflux. After the reaction was complete, the mixture was cooled to room temperature, concentrated under reduced pressure, and then ground in EtOH to obtain compound POS-D1a (17.539 g, 96%).

[1104] 1 ¹H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 7.68 (d, J = 8.4 Hz, 2H), 6.78 (d, J = 8.8 Hz, 2H), 4.37 (s, 2H). EI-MS m / z: 431(M + ).

[1105] Preparation of compound POS-D1b

[1106] CS2 (45 mL, 749.32 mmol) and KOH (6.5 g, 115.28 mmol) were added to a solution of compound POS-D1a (17.54 g, 115.28 mmol) in EtOH (200 mL) and DMF (100 mL) under an N2 atmosphere. After stirring at 85°C for 18 hours, EA (500 mL) and H2O (500 mL) were added to the mixture and then acidified with 1 M HCl. The organic layer was washed with H2O (500 mL) and brine (500 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was ground with ether / hexane to yield compound POS-D1b (20.7 g, 93%).

[1107] 1 ¹H NMR (400 MHz, DMSO-d6) δ 10.44 (s, 1H), 7.72 (d, J = 8.4 Hz, 2H), 6.94 (d, J= 8.0 Hz, 2H). EI-MS m / z: 195(M + ).

[1108] Preparation of compound POS-D1c

[1109] A solution of compound POS-D1b (5 g, 25.75 mmol) was added dropwise to a solution of THF (100 mL), and Et3N (4.3 mL, 30.9 mmol) and MeI (1.76 mL, 28.33 mmol) were added dropwise at 0°C. After stirring for 10 minutes at 0°C, the mixture was heated to room temperature. Then, the mixture was stirred for 2 hours, diluted with EA (100 mL x 2), the organic layer was washed with H2O (100 mL) and brine (100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was ground with ether to yield compound POS-D1c (5.15 g, 96%).

[1110] 1 ¹H NMR (400 MHz, DMSO-d6) δ 7.80 (d, J = 8.4 Hz, 2H), 6.94 (d, J = 8.4 Hz, 2H), 2.74 (s, 3H). EI-MS m / z: 209 (M + ).

[1111] Preparation of compound POS-D1d

[1112] 70% m-CPBA (11.4 g, 46.11 mmol) was added to a solution of compound POS-D1c (3.2 g, 15.37 mmol) in EtOH (150 mL) under an N2 atmosphere at 0°C. After stirring at room temperature for 5 hours, additional 70% m-CPBA (11.4 g, 46.11 mmol) was added. The mixture was then stirred overnight at room temperature, quenched with H2O (500 mL) and saturated NaHCO3 (300 mL), and diluted with EA (500 mL x 2). The organic layer was washed with brine (300 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was ground to HX / EA = 1:1 (100 mL) to yield compound POS-D1d (3.2 mg, 89%).

[1113] 1 ¹H NMR (400 MHz, DMSO-d6) δ 7.95 (d, J = 8.8 Hz, 2H), 7.01 (d, J = 8.8 Hz, 2H), 3.69 (4s, 3H). EI-MS m / z: 241(M + ).

[1114] Preparation of compound POS-D1

[1115] NaH (2.6 g, 0.065 mmol) was added dropwise at 0°C to a solution of tetraethylene glycol (17.3 ml, 0.10 mol) in THF (50 mL). After stirring the mixture at 0°C for 1 hour, propargyl bromide (5.95 g, 0.05 mol) was added. The mixture was stirred overnight at room temperature, quenched with ice and water, and diluted with EA (100 mL x 2). The organic layer was washed with H2O (100 mL) and brine (100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was ground with ether to obtain 3,6,9,12-tetraoxapentadex-14-phosph-1-ol (5.87 g, 51%).

[1116] 1 ¹H NMR (400 MHz, CDCl₃) δ 4.21 (s, 2H), 3.73-3.66 (m, 14H), 3.59-3.61 (m, 2H), 2.60 (s, 1H), 2.42 (t, J = 2.4 Hz, 1H).

[1117] 3,6,9,12-tetraoxapentadex-14-in-1-ol (660 mg, 2.84 mmol) and compound D-4-5 (310 mg, 1.29 mmol) were dissolved in THF (8 mL) and DMF (0.8 mL), and PPh3 (667 mg, 2.58 mmol) was added. The mixture was cooled to 0°C. 2.2 M DEAD (1.17 mL, 2.58 mmol) was added to the mixture, and the mixture was stirred at 0°C for 3 hours. After the reaction was complete, EA (15 mL × 2) and distilled water (15 mL) were added to extract the organic layer, and the mixture was washed with brine (20 mL). The obtained organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to yield compound POS-D1 (205 mg, 30%).

[1118] EI-MS m / z: 455(M + ).

[1119] [Example 11] Preparation of compound Int-TG13

[1120]

[1121] A solution of (1R,2S)-(-)-norephedrine (1 g, 6.61 mmol) compound in DCM (5 mL) was added to TEA (0.92 mL, 6.61 mmol) and acetyl chloride (0.47 mL, 6.61 mmol) at 0 °C under an N2 atmosphere. After stirring at room temperature for 2 hours, the mixture was quenched with H2O (7 mL). The organic layer was extracted with DCM (2 x 8 mL), dried with anhydrous MgSO4, filtered, and concentrated. The residue was purified by column chromatography to produce the compound Int-TG13 (987 mg, 78%).

[1122] 1 H-NMR (400 MHz, CDCl3) δ 7.28 (m, 5H), 5.70 (m, 1H), 4.86 (s, 1H), 4.34-4.30 (m, 1H), 3.65 (m, 1H), 2.00(s, 3H), 1.01 (d, J = 7.2 Hz, 3H)

[1123] [Example 12] Preparation of Compound IntB-Q3

[1124]

[1125] Preparation of compound IntB-Q3-1

[1126] NaIO4 (18 mg, 0.081 mmol) was added at room temperature to a solution of PNU-1529682 (52 mg, 0.081 mmol) in MeOH (5 ml) / distilled water (3 mL). After stirring for 2 hours, the mixture was concentrated under reduced pressure to obtain the crude compound IntB-Q3-1 (51 mg, 99%). EI-MS m / z: 628 (M +1 ).

[1127] Preparation of compound IntB-Q3

[1128] Compound in dry DCM (5 mL) IntB-Q3-12-(dimethylamino)ethylamine (6.1 μL, 0.089 mmol), TEA (34 μL, 0.243 mmol), and TBTU (52 mg, 0.162 mmol) were added to a solution of (51 mg, 0.081 mmol) at room temperature. After stirring for 1 hour, the mixture was diluted with DCM (2 x 8 mL). The organic layer was washed with H2O (8 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain the compound IntB-Q3 (38 mg, 67%).

[1129] EI-MS m / z: 698 (M +1 ).

[1130] [Example 13] Preparation of Compound L-2

[1131]

[1132] Compound L...

Claims

Claim 1 Chemical formula ( I' Conjugate of ) or a pharmaceutically acceptable salt thereof: Here: CB is a targeting moiety; cb and n are each independently integers having a value from 1 to 20; and each DL is independently a chemical formula ( I" It is a device having the structure of ): ( I" Each Q is an activator connected to L' by O or N independently; Z' is, in each case, an independent chemical formula ( I" The structure of ) (CB) cb It is a linker connecting to; each L' is a spacer moiety attached to SO2 through a heteroatom selected from O, S, and N independently, and each L' is selected such that the cleavage of the bond between L' and SO2 promotes the cleavage of the bond between L' and Q to release an activator; each X is independently -O-, -C(R b )(R c )-, or -N(R c )-and;E is an integer with a value of 1, 2, or 3;Ar is a 6-membered aryl or 6-membered heteroaryl ring;Y' is - N(R a )-, -O-, or -S-; one or more Xs are located in an ortho or para relation with respect to Y' on Ar; TG, when cleaved, SO2 and (Q) q -(L') w It is an inducing agent that generates N, O, or S atoms capable of initiating the emission of; each q is an integer having a value from 1 to 20 independently; each w and x is an integer having a value of 0 or 1 independently; and each R a and R c is independently hydrogen or lower alkyl; and one or more R b Under the condition that Z', each R b is independently Z', hydrogen, or lower alkyl; or R b and R c They form a 3-5-membered ring together with the atoms to which they are attached; provided that when w is 0, q is 1. Claim 2 In paragraph 1, X is -C(R b )(R c )-phosphorus, conjugate, or a pharmaceutically acceptable salt thereof. Claim 3 In paragraph 1, Ar is phenyl or pyridyl, conjugate, or a pharmaceutically acceptable salt thereof. Claim 4 In paragraph 1, Ar is a 6-membered aryl, conjugate, or pharmaceutically acceptable salt thereof. Claim 5 In claim 1, each L' is a conjugate or a pharmaceutically acceptable salt thereof that is a spacer moiety attached to SO2 through an independently -O- heteroatom. Claim 6 In claim 1, one or more X and Y' are conjugates or pharmaceutically acceptable salts thereof that are ortho relative to each other on Ar. Claim 7 In paragraph 1, E is a conjugate or a pharmaceutically acceptable salt thereof. Claim 8 In paragraph 1, at least one (CB) cb Z' connected to includes at least two of the following: C 10 -C 100 A linear or branched, saturated or unsaturated alkylene moiety, a conjugate, or a pharmaceutically acceptable salt thereof: (i) at least one heteroatom selected from -NH-, -C(=O), -O-, -S-, and -P-; (ii) at least one heteroarylene; (iii) at least one amino acid moiety, sugar bond, peptide bond, or amide bond; and (iv) C1-C 20 Alkyl, C6-C 20 Aryl C1-C8 alkyl, -(CH2) s COOH, and -(CH2) p One or more substituents selected from the group consisting of NH2, wherein s is an integer with a value from 0 to 10, and p is an integer with a value from 1 to 10. Claim 9 In claim 1, the Z' connecting CB and Ar is (CH2) connected to each other in a linear chain by covalent bonding b , L c , (P 1 ) a , W a1 , W a2 , W a3 , W b1 , Y 1 , and Y 2 Linking agents containing groups, conjugates, or pharmaceutically acceptable salts thereof: where, W a1 , W a2 , and W a3 Each is independently -NH-, -C(O)-, or -CH2- and;W b1 is an amide bond or triazolilene;P 1 is an amide bond, an amino acid residue, or a peptide; L c is an alkylene and;Y 1 -(CH2) q -(CH2CH2X") o - or -(CH2) q -(X"CH2CH2X") o -and;X" is -O-, -S-, -NH- or -CH2- and;Y 2 is a single combination or a group selected from the following: and W b2 is an amide bond or triazolilene; a is 0 to 10; b, c, and d are each independently integers with a value of 1 to 10; and o and q are each independently integers with a value of 1 to 10. Claim 10 In claim 9, the above Z' connecting CB and Ar is the chemical formula ( A Linking agents of ), conjugates, or pharmaceutically acceptable salts thereof: Here: * is the attachment point for CB; and ** is the attachment point for Ar. Claim 11 In Paragraph 9, P 1 is the following, conjugate, or pharmaceutically acceptable salt thereof: or Here:R 12 is hydrogen, alkyl, amino acid side chain, -(CH2) s C(O)R 13 or -(CH2) p NR 14 R 15 and; p is an integer with a value from 1 to 10; s is an integer with a value from 0 to 10; and R 13 is OH or -NH(CH2) s' (X'''CH2CH2) s" Z"-(CB) m Igo;R 14 and R 15 Each independently hydrogen or -C(O)(CH2) s' (X'''CH2CH2) s" Z"-(CB) m and; s'' is an integer with a value from 0 to 10; s' is an integer with a value from 1 to 10; m is an integer with a value of 0 or 1; X''' is -O-, -S-, -NH-, or -CH2-; and Z" is CB for R 14 or R 15 It is a linker that connects to the rest of; or Z" is a linker containing a reactive group. Claim 12 In paragraph 1, TG is a conjugate or a pharmaceutically acceptable salt thereof selected from the following: and Here: each R 21 is independently hydrogen or acetyl; and R 22 is hydrogen or a lower alkyl. Claim 13 In claim 1, TG is a conjugate or a pharmaceutically acceptable salt thereof selected from -NO2, -C(O)-(CH2)2C(O)-alkyl, and nitrobenzyl. Claim 14 In claim 1, Q is a conjugate or a pharmaceutically acceptable salt thereof, which is a drug selected from cytokines, immunomodulatory compounds, anticancer agents, antiviral agents, antibacterial agents, antifungal agents, antiparasitic agents, or combinations thereof. Claim 15 In paragraph 1, (Q) q -(L') w - is a conjugate or a pharmaceutically acceptable salt thereof selected from the following: Here:X 1 -O- or -NR a - and;X 2 and X 4 Each is independently absent, or -O-, -C(O)-, or -C(O)O- and;X 3 is -OC(=O)- and;w' is an integer with a value of 1, 2, 3, 4, or 5 and;R 9 and R 10 Each is independently hydrogen, alkyl, aryl, or heteroaryl, and said alkyl, aryl, and heteroaryl are unsubstituted or alkyl, -(CH2) u NH2, -(CH2) u NR u1 R u2 , and -(CH2) u SO2R u3 Substituted with one or more substituents selected from;R u1 , R u2 , and R u3 Each is independently hydrogen, alkyl, aryl, or heteroaryl; and u is an integer having a value from 1 to 10. Claim 16 In Paragraph 15, (Q) q -(L') w - is a conjugate or a pharmaceutically acceptable salt thereof selected from the following: Here, * is (Q) for -SO2- q -(L') w Indicates the attachment point of. Claim 17 In claim 1, the targeting moiety is a nanoparticle, immunoglobulin, nucleic acid, protein, oligopeptide, polypeptide, antibody, fragment of antigenic polypeptide, or lipibody, wherein the antibody is a conjugate or a pharmaceutically acceptable salt thereof selected from intact polyclonal antibodies, intact monoclonal antibodies, antibody fragments, single-stranded Fv (scFv) mutants, multispecific antibodies, bispecific antibodies, chimeric antibodies, humanized antibodies, human antibodies, fusion proteins comprising an antigenic determinant portion of an antibody, and other modified immunoglobulin molecules comprising an antigen recognition site. Claim 18 In claim 17, the above antibodies are Muromonab-CD3, Abxiximab, Rituximab, Daclizumab, Palivazumab, Infliximab, Trastuzumab (Herceptin), Etanercept, Basilixiammab, Gemtuzumab, Ozogamicin, Alemtuzumab, Ibritumomab, Tiuxetane, Adalimumab, Alefacept, Omalizumab, Epalizumab, Tocitumomab-I 131 , Cetuximab, Bevacizumab, Natalizumab, Ranibizumab, Panitumumab, Eculizumab, Rilonacept, Sertolizumab Pegol, Romiplostim, AMG-531, CNTO-148, CNTO-1275, ABT-874, LEA-29Y, Belimumab, TACI-Ig, Second-generation Anti-CD20, ACZ-885, Tocilizumab, Atlizumab, Mepolizumab, Pertuzumab, Humax CD20, Tremelimumab (CP-675 206), Tilcilimumab, MDX-010, IDEC-114, Inotuzumab Ozogamicin, HuMax EGFR, Aflibercept, HuMax-CD4, Ala-Ala, ChAglyCD3, TRX4, Catumaxomab, A conjugate or a pharmaceutically acceptable salt thereof selected from IGN101, MT-201, pregovomab, CH-14.18, WX-G250, AMG-162, AAB-001, motavizumab, MEDI-524, efumgumab, aurorap, lacivakumab, a third-generation anti-CD20, LY2469298, and veltuzumab. Claim 19 Chemical formula ( Ia Compound of ) or pharmaceutically acceptable salt thereof: Here: each Q is an activator independently connected to L' by O or N; Z' is independently absent in each case, or chemical formula ( Ia The structure of ) (CB) cb A linker connecting to, where CB is a targeting moiety and cb is an integer having a value from 1 to 20; each L' is a linker attached to SO2 through a heteroatom selected from O, S, and N independently, and each L' is selected such that the cleavage of the bond between L' and SO2 promotes the cleavage of the bond between L' and Q to release an activator; each X is independently -O-, -C(R b )(R c )-, or -NR c - and; E is an integer with a value of 1, 2, or 3; Ar is a 6-membered aryl or 6-membered heteroaryl ring; and Y' is -N(R a )-, -O-, or -S-; one or more Xs are located in an ortho or para relation with respect to Y' on Ar; TG, when cleaved, SO2 and (Q) q -(L') w It is an inducing agent that generates N, O, or S atoms capable of initiating the emission of; each q is an integer having a value from 1 to 20 independently; each w and x is an integer having a value of 0 or 1 independently; and each R a and R c is independently hydrogen or lower alkyl; and one or more R b Under the condition that Z', each R b is independently Z', hydrogen, or lower alkyl; or R b and R c They form a 3-5-membered ring together with the carbon atom to which they are attached; provided that when w is 0, q is 1. Claim 20 Chemical formula ( Ia Compound of ) or pharmaceutically acceptable salt thereof: Here: each Q is an activator independently connected to L' by O or N; Z' is a reactive group; each L' is an independently connecting group attached to SO2 through a heteroatom selected from O, S, and N, and each L' is selected such that the cleavage of the bond between L' and SO2 facilitates the cleavage of the bond between L' and Q to release the activator; each X is independently -O-, -C(R b )(R c )-, or -NR c - and; E is an integer with a value of 1, 2, or 3; Ar is a 6-membered aryl or 6-membered heteroaryl ring; and Y' is -N(R a )-, -O-, or -S-; one or more Xs are located in an ortho or para relation with respect to Y' on Ar; TG, when cleaved, SO2 and (Q) q -(L') w It is an inducing agent that generates N, O, or S atoms capable of initiating the emission of; each q is an integer having a value from 1 to 20 independently; each w and x is an integer having a value of 0 or 1 independently; and each R a and R c is independently hydrogen or lower alkyl; and one or more R b Under the condition that Z', each R b is independently Z', hydrogen, or lower alkyl; or R b and R c They form a 3-5-membered ring together with the carbon atom to which they are attached; provided that when w is 0, q is 1. Claim 21 A method for manufacturing a conjugate comprising reacting the compound of claim 20 with a targeting moiety, wherein the targeting moiety is a nanoparticle, immunoglobulin, nucleic acid, protein, oligopeptide, polypeptide, antibody, fragment of antigenic polypeptide, or lipidobody. Claim 22 An imaging composition comprising a conjugate of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof. Claim 23 A sensor compound comprising a conjugate of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof. Claim 24 A composition for use in detection comprising contacting a substance with a sensor compound, wherein the composition comprises a conjugate of any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof. Claim 25 delete Claim 26 delete Claim 27 delete Claim 28 delete Claim 29 delete Claim 30 delete Claim 31 delete Claim 32 delete Claim 33 delete Claim 34 delete Claim 35 delete Claim 36 delete Claim 37 delete Claim 38 delete Claim 39 delete Claim 40 delete Claim 41 delete Claim 42 delete Claim 43 delete Claim 44 delete Claim 45 delete Claim 46 delete Claim 47 delete Claim 48 delete Claim 49 delete Claim 50 delete Claim 51 delete Claim 52 delete Claim 53 delete Claim 54 delete Claim 55 delete Claim 56 delete Claim 57 delete Claim 58 delete Claim 59 delete Claim 60 delete Claim 61 delete Claim 62 delete Claim 63 delete Claim 64 delete Claim 65 delete Claim 66 delete Claim 67 delete Claim 68 delete Claim 69 delete Claim 70 delete Claim 71 delete Claim 72 delete

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