Conjugates, cell-binding molecule-drug conjugates containing the conjugates, and methods of using and making the conjugates and conjugates

Abstract

Linkers are provided that are used to conjugate cytotoxic agents to cell-binding molecules. The present invention provides linkers having a propiolyl, substituted acryloyl, or disubstituted propanoyl group, and the use of such linkers for attaching compounds, particularly cytotoxic agents, to cell-binding molecules.

Description

[Technical Field]

[0001] The present invention relates to a process for the conjugation of compounds, particularly cytotoxic agents, to cell-binding molecules. Linkers containing propiolyl, substituted acryloyl, or disubstituted propanoyl groups The present invention also relates to a method for the preparation of a drug by first modifying the drug with these conjugates, and Reacting with cell-binding molecules or first modifying cell-binding molecules with these conjugates, The resulting conjugate-drug assembly is then reacted with a drug or synthesized and the cell-binding molecule is then Cell-binding molecule-drug (cytotoxic agent) in specific manners, including by direct conjugation with Methods for preparing the conjugates are also disclosed. [Background technology]

[0002] Chemotherapy drugs usually cannot distinguish between normal and malignant cells, so therapeutic concentrations Therefore, it is likely to cause side effects that limit the tolerated dose below the clinically effective dose. In contrast, immunotherapy, usually in the form of monoclonal antibodies, is a major challenge. They can specifically bind to certain proteins or molecules in cells, leaving normal cells unharmed. Therefore, they have fewer side effects and a larger therapeutic window than chemotherapy drugs. Antibody-drug conjugates (ADCs) are highly potent cytotoxic drug payloads for the targeted treatment of cancer. It is a type of immunotherapy that combines tumor-specific binding monoclonal antibodies conjugated with This approach is being developed using the US FDA-approved drug Adcetris (brentuximab medoxomil). Treatment of Hodgkin's lymphoma with the US FDA-approved drug "Kadcyla" (ad- Trastuzumab emtansine has shown promising activity in the treatment of HER-2-positive breast cancer. Over the past two decades, both academia and the pharmaceutical industry have increasingly invested time and money in ADCs. With over 50 ADCs in clinical trials, the pharmaceutical industry expects to see another 8 ADCs in clinical trials within the next 2-3 years. It is predicted that ~10 ADC drugs may be approved for market (Non-Patent Documents 1-5) .

[0003] Many important parameters govern the successful development of antibody-drug conjugates for clinical use These include the selection of tumor target antigens that have limited expression on normal cells, and the selection of highly selective targets. Selective antibodies, which, when internalized and released, are required to induce highly potent target cell death. Key cytotoxic molecules, stable in circulation but releasing cytotoxic agents at target cells , a linker for bridging a cytotoxic molecule with an antibody, and a linker for attaching an antibody to a cytotoxic molecule. Although there have been many advances in ADC development, the off-target toxicity of ADCs remains a challenge. The mechanisms behind this are still poorly understood, and the duration of treatment in the clinic A significant number of ADCs that have completed the floor testing stage have a much narrower dosing regimen than preclinical models. The study is focused on dose-limiting toxicities (D-toxicities) that cannot always be predicted based on preclinical data. Therefore, the chemistry and design of ADCs have been hindered by the presence of LTs (Non-Patent Document 4). Research and development is currently moving beyond the efficacy of single payloads to include linker-payload components and conjugates. The scope of chemistry has expanded, particularly in the area of ​​target / disease-directed ADC conjugate-payload. Therefore, many drug developers and academic institutions and longer circulating half-lives, as well as improved batch-to-batch consistency in ADC manufacturing. Higher efficacy, reduced potential off-target toxicity, and in vivo efficacy of narrow spectrum ADCs A new and reliable method for site-specific ADC conjugation with potentially favorable biokinetics There is a strong focus on establishing

[0004] There are several approaches recently developed for the preparation of site-selective ADCs (non-patented). Reference 12). These include unpaired cysteines, e.g., THIOMA from Genentech. Introduction of a designed reactive cysteine ​​residue, designated B (Non-Patent Documents 13 and 14, Patent Document 14) References 2-6), Streptoverticillium mobaraense transglutaminase (mTG ) (Non-Patent Documents 10 and 15, Patent Document 7 (Rinat-Pfizer)) or microbial transglutaminase MTGase (Non-Patent Document 16, Patent Document 8 (Innate Pharma), Patent Document 9 (Bio-Ker Srl (IT) ... introduction of unnatural amino acids by mutagenesis (Non-Patent Document 18) ~21, Patent Documents 10-12 (Sutro Biopharm), Patent Documents 13-17 (Ambrx), Patent References 18-20 (Allozyne)), introduction of selenocysteine ​​into antibodies (Non-Patent Document 22, Patent Document Patent Reference 21 (US National Cancer Institute), Formylglycine synthase (FGE) The formylglycine (FGl) of the cysteine ​​located in the CXPXR consensus sequence was y) (Non-Patent Document 23, Patent Documents 22 to 26 (Redwood Bioscience)), and Glucoengineering using galactosyl and sialyltransferases These include the introduction of alkyl acids (Non-Patent Document 24, Patent Document 27 (Sanofi-Genzyme)). However, the above method requires reoptimization of the antibody engineering process and cell culture conditions. Therefore, bromo or dibromo maleimides, called next generation maleimides (NGMs), are (UCL Cancer Institute) or via a three-carbon bridge. Applying alkylating reagents (Non-Patent Document 26, Patent Documents 28 and 29 (PolyTherics Ltd) ) by recrosslinking the interchain disulfide bonds of reduced native antibodies. The conjugation method was actually used. We used bromomaleimide and dibromomaleimide linkers ( Patent Document 30), 2,3-disubstituted succinic acid / 2-monosubstituted / 2,3-disubstituted fumaric acid or Leic acid conjugates (Patent Documents 31 and 32), acetylenedicarboxylic acid conjugates (Patent Document 3 3 and 34), or hydrazine linkers (Patent Document 35), Several conjugation methods have been disclosed to recrosslink thiol pairs in which the sulfide bond has been reduced. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] International Publication No. WO2014 / 183555A1 [Patent Document 2] U.S. Patent No. 8,309,300 [Patent Document 3] U.S. Patent No. 7,855,275 [Patent Document 4] U.S. Patent No. 7,521,541 [Patent Document 5] U.S. Patent No. 7,723,485 [Patent Document 6] International Publication No. WO2008 / 141044 [Patent Document 7] U.S. Patent No. 8,871,908 [Patent Document 8] US Application No. 20130189287 [Patent Document 9] U.S. Patent No. 7,893,019 [Patent Document 10] U.S. Patent No. 8,778,631 [Patent Document 11] U.S. Patent Application No. 20100184135 [Patent Document 12] International Publication No. WO2010 / 081110 [Patent Document 13] International Publication No. WO2006 / 069246 [Patent Document 14] International Publication No. WO2007 / 059312 [Patent Document 15] U.S. Patent No. 7,332,571 [Patent Document 16] U.S. Patent No. 7,696,312 [Patent Document 17] U.S. Patent No. 7,638,299 [Patent Document 18] International Publication No. WO2007 / 130453 [Patent Document 19] U.S. Patent No. 7,632,492 [Patent Document 20] U.S. Patent No. 7,829,659 [Patent Document 21] U.S. Patent No. 8,916,159 [Patent Document 22] U.S. Patent No. 7,985,783 [Patent Document 23] U.S. Patent No. 8,097,701 [Patent Document 24] U.S. Patent No. 8,349,910 [Patent Document 25] U.S. Patent Application No. 20140141025 [Patent Document 26] U.S. Patent Application No. 20100210543 [Patent Document 27] U.S. Patent Application No. 20140294867 [Patent Document 28] International Publication No. WO2013 / 190272 [Patent Document 29] International Publication No. WO2014 / 064424 [Patent Document 30] International Publication No. WO2014 / 009774 [Patent Document 31] International Publication No. WO2015 / 155753 [Patent Document 32] International Publication No. WO2016 / 0596228 [Patent Document 33] International Publication No. WO2015 / 151080 [Patent Document 34] International Publication No. WO2016 / 0596228 [Patent Document 35] International Publication No. WO2015 / 151081 [Non-patent literature]

[0006] [Non-Patent Document 1] Lambert, JM Ther. Deliv. 2016, 7, 279-82 [Non-patent document 2] Jerjian, TV et al. Pharmacotherapy 2016, 36, 99-116 [Non-patent document 3] Donaghy, H. MAbs 2016, 8, 659-71 [Non-patent document 4] de Goeij, BE and Lambert, JM Curr Opin Immunol 2016, 40, 14-23 [Non-Patent Document 5] Mehrling, T. Future Oncol, 2015, 11, 549 [Non-patent document 6] Zhao, RY et al, 2011, J. Med. Chem. 54, 3606-23

Non-licensed Document 7

Non-licensed literature 9

Non-licensed literature 10

Non-licensed Document 11

Non-licensed Document 12

Non-licensed Document 13

Non-licensed Document 14

Non-licensed Document 15

Non-licensed Document 16

Non-licensed Document 17

Non-licensed Document 18

[0007] In this application, we extend the scope of our previous patent application. More importantly, The disulfur bridged linker of the present invention achieves a higher DAR (≧4). Conjugating two or more drugs per linker to a cell-binding molecule or two or more cells It is possible to conjugate at more than one thiol site on the cell-binding molecule. The main advantages of this application include: extending the half-life of the conjugate during targeted delivery. In order to increase the number of target cells exposed to the drug or effector, Two or more different functional molecules / drugs acting at different stages are conjugated together. Conjugating two or more cell-binding molecules for dual, triple, or multiple targeting strategies to cells Conjugation of functional molecules minimizes exposure to non-target cells, tissues, or organs. Precise control of drug payload and drug ratio at specific sites for a uniform final product.

[0008] The present invention relates to propiolyl, substituted acryloyl, or disubstituted propanoyl groups. and optionally further comprising a thiol-reactive group selected from the group consisting of phosphate amide, amine, hydrazine, triazoline, These include aryl, heteroaromatic, acetylamide, glycoside, and related groups, and are useful for drugs, and / or functional molecules and / or linkers for conjugation with cell-binding agents (e.g., antibodies) to provide. [Means for solving the problem]

[0009] In one embodiment of the invention, the linker is represented by formula (I) and (II): [ka]

[0010] During the ceremony, [ka] represents a single bond, and when bonded to a single bond or a double bond, [ka] may be an enantiomeric or stereoisomeric bond; [ka] represents either a single bond, a double bond, or a triple bond; [ka] represents a single bond, both Lv1 and Lv2 are not H; [ka] represents a double bond, either Lv1 or Lv2 can be H, but at the same time It will not become H; [ka] represents a triple bond, Lv1 is absent and Lv2 may optionally be H;

[0011] Lv1 and Lv2 represent the same or different leaving groups that can be displaced by a thiol; The leaving group may include, but is not limited to, a halide (fluoride, chloride, bromide, and and iodide), methanesulfonyl (mesyl), toluenesulfonyl (tosyl), trifluoromethylsulfonyl (triflate), trifluoromethylsulfonyl Nitrophenoxyl, N-succinimidyloxyl (NHS), phenoxyl; Nitrophenoxyl; Pentafluorophenoxyl, Tetrafluorophenoxyl, Trifluorophenoxyl Fluorophenoxyl, difluorophenoxyl, monofluorophenoxyl, pentacene Chlorophenoxyl, 1H-imidazol-1-yl, chlorophenoxyl, dichlorophenoxyl Noxyl, trichlorophenoxyl, tetrachlorophenoxyl, N-(benzotriazo 2-ethyl-5-phenylisoxazolium-3'-sulfonyl, Phenyloxadiazole-sulfonyl (-sulfone-ODA), 2-ethyl-5-phenyl Nylisoxazolium-yl, phenyloxadiazol-yl (ODA), oxadiazol-yl or an intermediate produced by a condensation reagent for the Mitsunobu reaction. ru;

[0012] Y reacts with cytotoxic agents to form disulfides, ethers, esters, thioethers, and thiol groups. Esters, peptides, hydrazones, carbamates, carbonates, amines (secondary, tertiary, or quaternary), imine, cycloheteroalkane, heteroaromatic, alkyl oxime, or a is a functional group capable of forming an amide bond;

[0013] Preferably, Y has the following structure: [ka] TIFF0007643734000009.tif98170

[0014] In the formula, X 1’ is F, Cl, Br, I, or Lv3; X 2’ are O, NH, N(R1 ), or CH2; R3 and R5 are each independently H, R1, aromatic, heteroaromatic Aromatic group, or one or more H atoms are -R1, -halogen, -OR1, -SR1, -NR substituted with -1R2, -NO2, -S(O)R1, -S(O)2R1, or -COOR1 Lv3 is an aromatic group; Lv4 is nitrophenol; N-hydroxysuccinimide (NHS ); Phenol; Dinitrophenol; Pentafluorophenol; Tetrafluorophenol difluorophenol; monofluorophenol; pentachlorophenol; Fluoride;Imidazole;Dichlorophenol;Tetrachlorophenol;1-Hydroxy Tosylate;Mesylate;2-Ethyl-5-phenylisoxazol azolium-3'-sulfonate, acid anhydrides formed with itself or with other acid anhydrides (e.g. For example, acetic anhydride, formic anhydride; or for peptide coupling reactions, or Mitsuno a leaving group selected from intermediates generated by condensation reagents for the BB reaction;

[0015] R1 is absent or C1-C8 (carbon number 1 to 8) alkyl; C2-C8 hetero alkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl; Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcycloalkyl, a alkylcarbonyl, heteroaryl; or C2-C8 (carbon number 2 to 8) ester, ester or a peptide containing 1 to 8 amino acids; or a peptide having the structural formula (OCH2 CH2) p or (OCH2CH(CH3)) p (p is an integer from 0 to about 1000) ethyleneoxy units, or combinations of the above groups;

[0016] Additionally, R1 may be C, N, O, S, Si, and Br, each of which is covalently bonded to Y and L1. and P, and preferably has 0 to 500 atoms. The atoms used are alkylene, alkenylene, alkynylene, ether, polyoxyethylene, and the like. alkylene, ester, amine, imine, polyamine, hydrazine, hydrazone, amide, Urea, semicarbazide, carbazide, alkoxyamine, alkoxyamine, urethane, Amino acids, peptides, acyloxyamines, or hydroxamic acids, or combinations thereof They may be bonded in any chemically relevant manner that forms a bond.

[0017] T is CH2, NH, NHNH, N(R3), N(R3)N(R 3’ ), O, S, C2- C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 Aryl, Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcyclo Alkyl, alkylcarbonyl, heteroaryl; peptides containing 1 to 4 amino acid units The amino acids are preferably aspartic acid, glutamic acid, arginine, hydroxybenzoate ... Styrene, lysine, serine, threonine, asparagine, glutamine, cysteine, selenium Cysteine, tyrosine, phenylalanine, glycine, proline, tryptophan, lanine; or one of the following structures:

[0018] [ka] TIFF0007643734000011.tif147164TIFF0007643734000012.tif30113

[0019] During the ceremony, [ka] is the linkage site;

[0020] X1, X2, X3, X4, X5, X6, X 1’ , X 2’ , and X 3’ are independent of each other. NH;NHNH;N(R3);N(R3)N(R 3’ );O;S;C1-C6 alkyl C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C 3-C8 aryl, Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkoxy or 1 to 8 amino acids wherein R and R 3’ are each independently H; C1-C8 alkyl; C2-C8 hetero alkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl; Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcycloalkyl, a alkylcarbonyl, heteroaryl; C1-C8 esters, ethers or amides; or structural formula (OCH2CH2) p or (OCH2CH(CH3)) p (p;0~about 1 000) or a combination of the above;

[0021] L1 and L2 are each independently the same or different and are selected from the group consisting of O, NH, S, NHNH, N( R3), N(R3)N(R 3’ ), formula (OCH2CH2) p OR3, or (OCH2 CH(CH3)) p OR3, or NH(CH2CH2O) p R3 or NH(C H2CH(CH3)O) p R3, or N[(CH2CH2O) p R3][(CH2C H2O) p’ R 3’ ], or (OCH2CH2) p COOR3 or CH2CH 2(OCH2CH2) pCOOR3 polyethyleneoxy unit (wherein p and p' are each independently an integer of 1 to about 1000), or a combination thereof; C2-C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl C3-C8 aryl, Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroaromatic alkylcycloalkyl, alkylcarbonyl, heteroaryl; wherein R and R 3’ teeth each independently H; C1-C8 alkyl; C2-C8 heteroalkyl, alkylcyclo Aryl, Ar-alkyl, heterocycloalkyl; C3-C8 aryl, Ar-alkyl, heterocycle, Carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, hetero aryl; C1-C8 ester, ether, or amide; 1 to 8 amino acids; or Structural formula (OCH2CH2) p or (OCH2CH(CH3)) p (p;0~about 100 0 integer), or a combination of the above;

[0022] L1 or L2 is 6-maleimidocaproyl (MC), maleimidopropanoyl (MP ), valine-citrulline (val-cit or vc), alanine-phenylalanine (a la-phe or af), p-aminobenzyloxycarbonyl (PAB), 4-thio- Pentanoic acid ester (SPP), 4-(N-maleimidomethyl)cyclohexane-1-carboxylate Carboxylic acid ester (MCC), (4-acetyl)aminobenzoic acid (SIAB), 4-thio -butyric acid ester (SPDB), 4-thio-2-hydroxysulfonyl-butyric acid ester (2 -sulfo-SPDB), or natural or unnatural amino acid units containing 1 to 8 natural or unnatural amino acid units may be composed of one or more linker moieties selected from non-naturally occurring peptides.

[0023] m1, m2, m3, m4, and m5 each independently represent 1 to 10, preferably 1 to 10. are integers from 0 to 4;

[0024] In another aspect, the present invention provides a compound represented by formula (III), (IV), (V), (VI), (VII), (VIII) or (IX) cell-binding agent-drug conjugate, Each Drug reacts at the end of the bridge linker:

[0025] [ka] TIFF0007643734000015.tif218136

[0026] where Cb, Cb', Cb''Cb'' are the same or different and are cell binding agents or immunoreactive agents. It represents an immunotherapeutic protein, preferably an antibody or antibody fragment;

[0027] Inside the right brackets (square brackets) of formulas (III), (VII), (VIII), and (IX) has a linker-drug moiety that is conjugated to a thiol pair on the cell-binding agent / molecule. The ester is preferably dithiothreitol (DTT), dithioerythritol (DTE), L-glutathione (GSH), tris(2-carboxyethyl)phosphine (TCEP) , 2-mercaptoethylamine (β-MEA), or / and β-mercaptoethanol (β The intrachain disulfide bonds are reduced by a reducing agent selected from the group consisting of 2-ME, 2-ME, and 2-ME. It is a pair of sulfur atoms.

[0028] Drug, Drug', and Drug'' are the same or different cytotoxic agents or treatments. To promote the binding or stabilization of drugs, or immunotherapeutic proteins, or cell-binding agents R1 represents a functional molecule or a cell surface receptor binding ligand, and R1 represents a bridge linker of the present invention. and R1 is a C1-C8 alkane; Alkylene, alkenylene, alkynylene, aromatic ring, ether, polyoxyalkylene, Esters, amines, imines, polyamines, hydrazines, hydrazones, amides, ureas, semi- Carbazide, carbazide, alkoxyamine, urethane, amino acid, peptide, acylo xyamines, hydroxamic acids, disulfides, thioethers, thioesters, carbamates a carbonate, a heterocycle, a heteroalkyl, a heteroaromatic, or an alkoxide; The drug may also include a cytotoxic molecule, an immunotherapeutic agent, or a combination thereof. compounds, chemotherapeutic compounds, antibodies or antibody fragments, siRNA or DNA molecules , or may be a cell surface-bound ligand;

[0029] [ka] represents a single or double bond;

[0030] Inside the square brackets, a pair of sulfur atoms on the cell-binding molecule binds the cell-binding molecule. There are drugs that are conjugated with children.

[0031] m1, m 1’ , m 1’’ , m2, m 2’ , m 2’’ , m3, m4, m5, m 4’ , m5 ’ , m 4’’ , m 5’’ , m 4’’’ , m 5’’’ , m4’’’’ , and m 5’’’’ , each independently represents an integer of 1 to 10, preferably 1 to 4;

[0032] X1, X 1’ , X 1’’ , X 1’’’ , and X 2’’’’ are each independently NH; NHNH;N(R3);N(R3)N(R 3’ );O;S;C1-C6 alkyl;C2- C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 Aryl, Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcycloa alkyl, alkylcarbonyl, heteroaryl; 1 to 8 amino acids; BiR 3’ are each independently H; C1-C8 alkyl; C2-C8 heteroalkyl; Aryl, cycloalkyl, heterocycloalkyl; C3-C8 aryl, Ar-alkyl , heterocycle, carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl C1-C8 esters, ethers, or amides; or CH2CH2) p or (OCH2CH(CH3)) p (p: an integer between 0 and approximately 1000) or a combination thereof, and further comprising X1, X 1’ , X 1’ ’ , X 1’’’ , and X 2’’’’ may each independently be absent;

[0033] R1, R 1’ , and R 1’ ' are the same or different and are C1-C8 alkyl; C2-C C3-C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; Aryl, Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcycloalkenyl alkyl, alkylcarbonyl, heteroaryl; C2-C8 esters, ethers or Amide; or structural formula (OCH2CH2) p or (OCH2CH(CH3)) p (p; an integer from 0 to about 1000) polyethyleneoxy units, or a combination of the above groups;

[0034] L1, L 1’ , L 1’’ , L 1’’’ , L2, L 2’ , L 2’’ , and L 2’’’ ,teeth The definitions of L1 and L2 in formula (I) and formula (II) are the same, and they are not the same at the same time. May;

[0035] n is 1 to 20; T is the same as previously described in formula (I).

[0036] In a further aspect, the present invention provides a modified cell-binding agent of formula (III), wherein The cell-binding agent Cb binds to the ATP via a pair of thiols generated by the reduction of disulfide bonds. and reacting with the bridge linker and having a functional group Y capable of reacting with a drug:

[0037] [ka] TIFF0007643734000018.tif218135

[0038] During the ceremony, [ka] Z1, Z2, n, R1, R2, m1, m2, X1, and X2 are represented by the formulas (I) and (II). It is the same definition; [ka] and Cb have the same definition as in formulae (III) to (IX);

[0039] In yet another aspect, the present invention provides modified drugs of formula (XVII) and (XVIII): wherein the drug "Drug" reacts with the linker of formula (I) and (II) and still Substituted acrylic or propiolic groups (e.g., hydroxyl groups) capable of reacting with a pair of thiols of the cell-binding agent. It has a thiol-reactive group (propiolic group).

[0040] [ka]

[0041] During the ceremony, [ka] L1, L2, R1, T, m1, m2, m3, m4, m5, X1, Lv1, and Lv2 are Drug 1 has the same definition as in formula (I). Drug 1 has the same definition as in formula (II).

[0042] The present invention further relates to a method for preparing the cell-binding molecule-drug conjugates of formulas (III)-(IX). , where the drug "Drug" is bound to the cell-binding agent via a cross-linker.

[0043] The present invention also relates to methods for preparing modified cell-binding molecules of formula (X)-(XVI), The cell-binding molecule reacts with the linkers of formula (I) and (II).

[0044] The present invention also relates to a method for preparing modified drugs of formula (XVII) and (XVIII), The drug reacts with the bridge linkers of formulas (I) and (II). [Brief explanation of the drawings]

[0045] [Figure 1] The synthesis of the present conjugates containing two or four drugs and their application in conjugation to antibodies via a pair of thiols is presented. [Figure 2] The synthesis of the present conjugates containing two or four drugs and their application in conjugation to antibodies via a pair of thiols is presented. [Figure 3] The synthesis of the present conjugate containing a drug and polyethylene glycol and its application in conjugation with an antibody via a pair of thiols is presented. [Figure 4] The synthesis of the present drug-containing conjugates and their application in conjugation with antibodies via a pair of thiols is shown. [Figure 5] The synthesis of the present conjugates comprising a drug, an amino acid, and polyethylene glycol, and the application of the conjugates in conjugation with antibodies via a pair of thiols are presented. [Figure 6] The synthesis of a conjugate containing a drug, phosphamide, and polyethylene glycol and its application in conjugation with an antibody via a pair of thiols is presented. [Figure 7] The synthesis of a conjugate containing a drug and a phosphamide and its application in conjugation with an antibody via a pair of thiols is presented. [Figure 8] The synthesis of a conjugate containing a drug and a phosphamide and its application in conjugation with an antibody via a pair of thiols is presented. [Figure 9] The synthesis of the present conjugate containing a drug and polyethylene glycol and its application in conjugation with an antibody via a pair of thiols is presented. [Figure 10] The synthesis of the present conjugates containing a drug and conjugate moieties L1 and L2, and the application of the conjugates in conjugation with antibodies via a pair of thiols, are shown. [Figure 11] 1 shows the synthesis of a conjugate of the present invention containing a prostate surface antigen (PSA) binding ligand. [Figure 12] The synthesis of the present conjugates containing prostate surface antigen (PSA) binding ligands and the application of the conjugates in conjugation with antibodies via a pair of thiols is shown. [Figure 13] 1 shows the synthesis of intermediates for tubulysin analogues. [Figure 14] 1 shows the synthesis of antibody-tubulysin analog conjugates via conjugable tubulysin analogs and the present linkers. [Figure 15] 1 shows the synthesis of antibody-MMAF analog conjugates via the present linker. [Figure 16] 1 shows the synthesis of antibody-MMAF analog conjugates via the present linker. [Figure 17] 1 shows the synthesis of antibody-MMAF analog conjugates via the present linker. [Figure 18] 1 shows the synthesis of antibody-MMAF analog conjugates via the present linker. [Figure 19] 1 shows the synthesis of antibody-tubulysin analog conjugates via tubulysin analog moieties and the present linkers. [Figure 20] 1 shows the synthesis of antibody-tubulysin analog conjugates via the present linker. [Figure 21] 1 shows the synthesis of antibody-tubulysin analog conjugates via the present linker. [Figure 22] 1 shows the synthesis of antibody-tubulysin analog conjugates via the present linker. [Figure 23] 1 shows the synthesis of antibody-tubulysin analog conjugates via the present linker. [Figure 24] 1 shows the synthesis of antibody-tubulysin analog conjugates via the present linker. [Figure 25] 1 shows the synthesis of a conjugate containing both an MMAF analog and a tubulysin analog via the present linker. [Figure 26]1 shows the synthesis of a conjugate containing both an MMAF analog and a PBD dimer analog via the present linker. [Figure 27] 1 shows the synthesis of a conjugate containing both an MMAF analog and a PBD dimer analog via the present linker. [Figure 28] 1 shows the synthesis of a conjugate containing two MMAF analogs via the present linker. [Figure 29] 1 shows the synthesis of antibody-tubulysin analog conjugates via the present linker. [Figure 30] 1 shows the synthesis of antibody-tubulysin analog conjugates via the present linker. [Figure 31] 1 shows the synthesis of antibody-tubulysin analog conjugates via the present linker. [Figure 32] 1 shows the synthesis of antibody-tubulysin analog conjugates via the present linker. [Figure 33] 1 shows the synthesis of antibody-tubulysin analog conjugates via the present linker. [Figure 34] 1 shows the synthesis of antibody-tubulysin analog and conjugable MMAF analog conjugates via the present linker. [Figure 35] 1 shows the synthesis of antibody-MMAF analog conjugates via the present linker. [Figure 36] 1 shows the synthesis of antibody-amatoxin analog conjugates via the present linker. [Figure 37] 1 shows the synthesis of antibody-amatoxin analog conjugates via the present linker. [Figure 38] 1 shows the synthesis of antibody-amatoxin analog conjugates via the present linker. [Figure 39] 1 shows the synthesis of antibody-amatoxin analog conjugates via the present linker. [Figure 40] 1 shows the synthesis of antibody-tubulysin analog and antibody-MMAF analog conjugates via the present linkers. [Figure 41] 1 shows the synthesis of antibody-tubulysin analog conjugates via the present linker. [Figure 42] 1 shows the synthesis of antibody-tubulysin analog, antibody-PBD dimer analog, and antibody-MMAF analog conjugates via the present linkers. [Figure 43] 1 shows the synthesis of antibody-tubulysin analogue conjugates containing PMSA-binding ligands and antibody-tubulysin analogue conjugates containing PEG chains via the present linkers. [Figure 44] An SDS-PAGE gel containing the developing reducing agent DTT is shown. Lanes 1 and 11 are biomarkers, lanes 2 and 16 are conjugate (232), lanes 3 and 15 are conjugate (339), lane 4 is conjugate (234), lane 5 is conjugate (238), lane 6 is conjugate (261), lanes 7 and 17 are conjugate (308), lane 8 is conjugate (239), lane 9 is conjugate (476), lane 10 is conjugate (478), lane 12 is conjugate (360), lane 14 is conjugate (238), lane 18 is conjugate (481), lane 19 is conjugate (483), and lane 20 is T-DM1. Conjugates (232), (234), (238), (261), (308), (339), (354), and (360) via our cross-linker exhibited a major 75 KD band, indicating that the heavy and light chains of the mAb were cross-linked with the linker. However, the bond between the two heavy chains of these conjugates could be displaced with the reducing agent DTT, resulting in a faint 150 KD band. In SDS-PAGE, the cross-links of conjugates (476), (478), (481), and (483) were also displaced with DTT, and the 75 KD and 150 KD bands were also very faint (reversible conjugation). In comparison, non-cross-linked T-DM1 did not exhibit a 75 KD band, and conjugate (239) prepared without UV light exhibited a faint 75 KD band, indicating that it may not be cross-linked under the conjugation conditions. [Figure 45]The antitumor efficacy of conjugates 232, 308, 327, 339, 476, 485, and 500 was compared with that of T-DM1 in a human gastric tumor N87 cell model. Conjugates 232, 308, 327, 339, 476, and 485 were administered intravenously at a dose of 5 mg / kg, while conjugates 339 and 500 were administered intravenously at a dose of 4 mg / kg. The seven conjugates tested here demonstrated superior antitumor activity to T-DM1. In the groups treated with compounds 476, 483, 339, and 500, all 6 / 6 animals were free of measurable tumors from days 14 to 52. In contrast, T-DM1 at a dose of 5 mg / kg failed to eliminate tumors and only inhibited tumor growth for 31 days. Conjugates 232, 308, and 327 did not completely eradicate tumors at a dose of 5 mg / kg. DETAILED DESCRIPTION OF THE INVENTION

[0046] definition

[0047] "Alkyl" means an alkyl group formed by removing one or two hydrogen atoms from a carbon atom. It refers to an aliphatic hydrocarbon group or monovalent group derived from a carboxylic acid. The alkyl group may be linear or branched, having one or more carbon atoms. is a group having multiple lower C alkyl groups, such as methyl, ethyl, or propyl groups, attached thereto. Specific examples of alkyl groups include methyl, ethyl, n-propyl, and i-propyl. butyl, n-butyl, t-butyl, n-pentyl, 3-pentyl, octyl, nonyl, decyl , cyclopentyl, cyclohexyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl , 2,2-dimethylpentyl, 2,3-dimethylpentyl, 3,3-dimethylpentyl, 2,3,4-trimethylpentyl, 3-methylhexyl, 2,2-dimethylhexyl, 2 ,4-dimethylhexyl, 2,5-dimethylhexyl, 3,5-dimethylhexyl, 2, 4-dimethylpentyl, 2-methylheptyl, 3-methylheptyl, n-heptyl, iso Heptyl, n-octyl, and isooctyl are included. C1-C8 alkyl groups are unsubstituted. and may be substituted with one or more substituents, including but not limited to the following: The substituents include -C1-C8 alkyl, -O-(C1-C8 alkyl), aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH2, -C( O)NHR', -C(O)N(R')2, -NHC(O)R', -SR', -S(O)2 R', -S(O)R', -OH, -halogen, -N3, -NH2, -NH(R'), -N (R')2 and -CN, where R' is independently C1-C8 alkyl and aryl.

[0048] "Halogen" refers to a fluorine, chlorine, bromine, or iodine atom, and includes fluorine and chlorine atoms. is preferred.

[0049] "Heteroalkyl" means a group of 1 to 4 carbon atoms independently selected from the group consisting of O, S, and N. "C2-C8 alkyl substituted at a heteroatom selected from:

[0050] "Carbocycle" means a monocyclic ring system having 3 to 8 carbon atoms or a monocyclic ring system having 7 to 13 carbon atoms. refers to a bicyclic saturated or unsaturated ring. Monocyclic carbocyclic rings have 3 to 6 rings, more typically 5 or 6 Bicyclic carbocycles have 7 to 12 ring atoms and are bicyclic [4,5], [ Arranged as [5,5], [5,6], or [6,6], or 9-10 ring atoms and arranged as a bicyclic ring system [5,6] or [6,6]. Rings (C3~C8 carbocycles) include -cyclopropyl, -cyclobutyl , -cyclopentyl, -cyclopentadienyl, -cyclohexyl, -cyclohexenyl , -1,3-cyclohexadienyl, -1,4-cyclohexadienyl, -cycloheptyl -1,3-cycloheptadienyl, -1,3,5-cycloheptatrienyl, -cycloheptatrienyl Includes, but is not limited to, -cyclooctyl, and -cyclooctadienyl.

[0051] C3-C8 carbocycles can have 3, 4, 5, 6, 7, or refers to an 8-membered saturated or unsaturated non-aromatic carbocyclic compound. C3-C8 carbocyclic rings are unsubstituted It may be substituted with one or more substituents, such as Examples include, but are not limited to, -C1-C8 alkyl, -O-(C1-C8 alkyl), -aryl -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH2, -C (O)NHR', -C(O)N(R')2, -NHC(O)R', -SR', -S(O)R ', -S(O)2R', -OH, -halogen, -N3, -NH2, -NH(R'), -N (R')2, and -CN, where each R' is independently C1 to C8 alkyl and and aryl.

[0052] "Alkenyl" refers to a straight-chain alkyl group having 2 to 8 carbon atoms in the chain and containing a carbon-carbon double bond. Alkenyl groups include, for example, ethenyl, propanol, and the like. n-butenyl, i-butenyl, 3-methylbut-2-enyl, n-pentenyl , hexylenyl, heptenyl, and octenyl.

[0053] "Alkynyl" refers to a straight-chain alkyl group having 2 to 8 carbon atoms in the chain and containing a carbon-carbon triple bond. The alkynyl group refers to an aliphatic hydrocarbon group which may be cyclic or branched. Examples of the alkynyl group include ethynyl, Propynyl, n-butynyl, 2-butynyl, 3-methylbutynyl, 5-pentynyl, n- Includes pentynyl, hexynyl, heptynyl, and octynyl.

[0054] An "alkylene" is a group formed by attaching two hydrogen atoms to the same or two different carbon atoms of a parent alkane. a saturated, C1-18, monovalent radical center derived by removing It refers to a straight or branched chain or cyclic hydrocarbon group. Typical alkylene groups include methylene ( -CH2-), 1,2-ethyl (-CH2CH2-), 1,3-propyl (-CH2CH 2CH2-), 1,4-butyl (-CH2CH2CH2CH2-), etc. Not limited to:

[0055] "Alkenylene" refers to a group that has two hydrogen atoms attached to the same or two different carbon atoms of a parent alkene. Unsaturated carbon atoms having 2 to 18 carbon atoms and two monovalent radical centers derived by removing a carbon atom Typical alkenylene groups include 1, Including, but not limited to, 2-ethylene (-CH=CH-).

[0056] An "alkynylene" is an alkylene group that is formed by attaching two hydrogen atoms to the same or two different carbon atoms of a parent alkyne. Unsaturated carbon atoms having 2 to 18 carbon atoms and two monovalent radical centers derived by removing a carbon atom Typical alkynylene groups include acetonitrile, ... Examples include, but are not limited to, ethylene, propargyl, and 4-pentynyl.

[0057] "Aryl" or Ar refers to an alkyl group having 3 to 14 carbon atoms, preferably 6 to 10 carbon atoms. "Heteroaromatic group" refers to an aromatic or heteroaromatic group consisting of one or several rings, including The term refers to a group in which some carbons, preferably 1, 2, 3, or 4 carbon atoms on the aromatic group are O , N, Si, Se, P, or S, preferably O, S, and N. The term aryl or Ar also refers to a group in which one or several H atoms are independently -R', -halogen, -OR', or -SR', -NR'R'', -N=NR', -N=R', -NR'R'' , -NO2, -S(O)R', -S(O)2R', -S(O)2OR', -OS(O)2 OR', -PR'R'', -P(O)R'R'', -P(OR')(OR''), -P( Replaced by -OP(O)(OR')(OR'') or -OP(O)(OR')(OR'') The R' and R'' are independently H, alkyl, alkenyl, alkynyl, The alkyl group may be a heteroalkyl, aryl, arylalkyl, carbonyl, or a pharmaceutical salt thereof.

[0058] A "heterocycle" is a ring in which 1 to 4 ring carbon atoms are independently selected from O, N, It refers to ring systems substituted with heteroatoms from the group of S, Se, B, Si, and P. The most common heteroatoms are O, N, and S. Heterocycles are described in The Handbook of Chemistry and Ph , 78th Edition, CRC Press, Inc., 1997-1998, pp. 225-226, and the disclosure thereof are incorporated herein by reference. Preferred non-aromatic heterocycles include, but are not limited to: However, epoxy, aziridinyl, thiranyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl Lysinyl, oxiranyl, tetrahydrofuranyl, dioxolanyl, tetrahydropyranyl dioxanyl, dioxolanyl, piperidyl, piperazinyl, morpholinyl, pyranilinyl Imidazolinyl, pyrrolinyl, pyrazolinyl, thiazolidinyl, tetrahydropyranyl dihydropyranyl, tetrahydropyridyl, dihydropyridyl, tetrahydropyriminyl dihydrothiopyranyl, dihydrothiopyranyl, azepanyl, and condensation resulting from condensation with a phenyl group The system is included.

[0059] The term "heteroaryl" or aromatic heterocycle refers to a 3- to 14-membered, preferably 5- to 10-membered, aromatic heterocycle. Aromatic heterocyclic, monocyclic, bicyclic, or polycyclic rings, examples of which include pyrrolyl, pyridyl, and the like. , pyrazolyl, thienyl, pyrimidinyl, pyrazinyl, tetrazolyl, indolyl, quino Linyl, purinyl, imidazolyl, thienyl, thiazolyl, benzothiazolyl, furanyl , benzofuranyl, 1,2,4-thiadiazolyl, isothiazolyl, triazolyl, tetrazolyl pyrazolyl, isoquinolyl, benzothienyl, isobenzofuryl, pyrazolyl, carbazolyl benzimidazolyl, isoxazolyl, pyridyl-N-oxide, and phenyl groups This includes condensed systems resulting from the condensation of

[0060] "Alkyl," "cycloalkyl," "alkenyl," "alkynyl," "aryl" "Heteroaryl", "heterocyclic" etc. have two hydrogen atoms. The corresponding "alkylene," "cycloalkylene," and "cycloalkylene" groups are formed by removing atoms. ", "alkenylene", "alkynylene", "arylene", "heteroarylene", " It also refers to heterocyclic groups, etc.

[0061] "Arylalkyl" refers to an alkyl group consisting of a carbon atom, typically a terminal or sp 3 Water bonded to a carbon atom It refers to an acyclic alkyl group in which one of the carbon atoms is substituted with an aryl group. The alkyl groups include, but are not limited to, benzyl, 2-phenylethan-1-yl , 2-phenylethen-1-yl, naphthylmethyl, 2-naphthylethan-1-yl, 2 -naphthylethen-1-yl, naphthobenzyl, 2-naphthophenylethan-1-yl, etc. Includes:

[0062] "Heteroarylalkyl" refers to a heteroaryl group consisting of a carbon atom, typically a terminal or sp 3 Attached to a carbon atom It refers to an acyclic alkyl group in which one of the hydrogen atoms is replaced by a heteroaryl group. Typical heteroarylalkyl groups include 2-benzimidazolylmethyl, 2-furylethy Contains rules.

[0063] Examples of "hydroxy protecting groups" include methoxymethyl ether, 2-methoxyethoxymethyl ether, tetrahydropyranyl ether, benzyl ether, p-methoxybenzyl ether, trimethylsilyl ether, triethylsilyl ether, triisopropylsilyl ether silyl ether, t-butyldimethylsilyl ether, triphenylmethylsilyl ether , acetate esters, substituted acetate esters, pivaloates, benzoates, methanesulfonates , and p-toluenesulfonate.

[0064] A "leaving group" refers to a functional group that can be displaced by another functional group. Suitable leaving groups are well known in the art and include, for example, halides (e.g., chloride). , bromide, and iodide), methanesulfonyl (mesyl), p-toluenesulfonyl (thiazolinone), trifluoromethylsulfonyl (triflate), and trifluoromethylsulfonyl Preferred leaving groups include nitrophenol; N-hydroxysuccinate; Cinimide (NHS); Phenol; Dinitrophenol; Pentafluorophenol; Tetrafluorophenol; Difluorophenol; Monofluorophenol; Pentak Triflate; Imidazole; Dichlorophenol; Tetrachlorophenol 1-Hydroxybenzotriazole;Tosylate;Mesylate;2-Ethyl-5 -phenylisoxazolium-3'-sulfonate, formed with itself or with other acid anhydrides acid anhydrides (e.g., acetic anhydride, formic anhydride); or for peptide coupling reactions or an intermediate formed by a condensation reagent for the Mitsunobu reaction.

[0065] The following abbreviations may be used herein and have the definitions indicated below: Boc, tert-Butoxycarbonyl; BroP, Bromotrispyrrolizinophosphonium hexa Fluorophosphate; CDI, 1,1'-carbonyldiimidazole; DCC, dicyclohexyl Dihexylcarbodiimide; DCE, 1,2-dichloroethane; DCM, dichloromethane ;DIAD, diisopropyl azodicarboxylate;DIBAL-H, hydrogenated diisobutyl aldehyde Aluminum; DIPEA, diisopropylethylamine; DEPC, diethylphosphoric acid didiate; DMA, N,N-dimethylacetamide; DMAP, 4-(N,N-dimethyl (amino)pyridine; DMF, N,N-dimethylformamide; DMSO, dimethyl sulfoxide dithioester; DTT, dithioester; EDC, 1-(3-dimethylaminopropyl)-3 -ethylcarbodiimide hydrochloride; ESI-MS, electrospray mass spectrometry; HATU , O-(7-azabenzotriazol-1-yl)-N,N,N'-N'-tetramethyl Uronium hexafluorophosphate; HOBt, 1-hydroxybenzotriazole; H PLC, high-pressure liquid chromatography; NHS, N-hydroxysuccinimide; MMP , 4-methylmorpholine; PAB, p-aminobenzyl; PBS, phosphate-buffered saline (pH 7.0-7.5); PEG, polyethylene glycol; SEC, size exclusion chromatography TCEP, tris(2-carboxyethyl)phosphine; TFA, triflate Fluoroacetic acid; THF, tetrahydrofuran; Val, valine.

[0066] "Amino acids" can be natural and / or unnatural amino acids, preferably α-amino acids Natural amino acids are those encoded by the genetic code and include alanine, arginine, Asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, Styridine, isoleucine, leucine, lysine, methionine, phenylalanine, proline , serine, threonine, tyrosine, tryptophan, and valine. are derived forms of protein-forming amino acids, e.g., hydroxyproline, lanthionine 2-aminoisobutyric acid, dehydroalanine, γ-aminobutyric acid (neurotransmitter), ornithin Citrulline, β-alanine (3-aminopropanoic acid), γ-carboxyglutamate Selenocysteine ​​(present as in most eukaryotes, but not directly linked to DNA) (not present in some archaea and one bacterium), pyrrolysine (only present in some archaea and one bacterium), N-phosphatase methylmethionine (often the first amino acid in proteins in bacteria, mitochondria, and chloroplasts) amino acid), 5-hydroxytryptophan, L-dihydroxyphenylalanine, triiodothyronine Dothyronine, L-3,4-dihydroxyphenylalanine (DOPA), and O-phosphoric acid The term amino acid also includes amino acid analogs and mimetics. is the same as that of a natural amino acid, except that the R group is not one found in a natural amino acid. These are compounds with the same general structure: H2N(R)CHCO2H. Examples of analogs include Moselin, norleucine, methionine sulfoxide, and methionine methyl sulfonium Preferably, the amino acid mimetic has a structure different from the general chemical structure of an α-amino acid. The term "unnatural amino acid" refers to a compound that has the structure, but functions similarly to, It is intended to represent the "D" stereochemical form, whereas naturally occurring amino acids are in the "L" form. When 1 to 8 amino acids are used in the present application, the amino acid sequence is selected from the cleavage recognition sequences of proteases. Many cleavage recognition sequences are known in the art, for example, Matayoshi et al. Science 247: 954 (1990); Dunn et al. Meth. Enzymol. 241: 25 4 (1994);Seidah et al. Meth. Enzymol. 244: 175 (1994);Thornberry, Meth. Enzymo l. 244: 615 (1994);Weber et al. Meth. Enzymol. 244: 595 (1994);Smith et al. Me th. Enzymol. 244: 412 (1994); and Bouvier et al. Meth. Enzymol. 248: 614 (1995) See, e.g., AA-Val, the disclosure of which is incorporated herein by reference. In particular, the sequence , Ala-Ala, Val-Val, Val-Ala-Val, Lys-Lys, Ala-Asn-Val, Val-Leu-Lys, Cit-Cit, Val -Lys, Ala-Ala-Asn, Lys, Cit, Ser, and Glu.

[0067] A "glycoside" is a compound in which a sugar group is bonded to another group via its anomeric carbon through a glycosidic bond. Glycosides are molecules that are linked together. Glycosides are O-(O-glycosides), N-(glycosylamines), ), S-(thioglycoside), or C-(C-glycoside) glycosidic bonds The core empirical formula is C m (H2O) n (where m is different from n, m and n are <36), where glycosides include glucose (dextrose ), fructose (levulose), allose, altrose, mannose, gulose, yogurt Dose, galactose, talose, galactosamine, glucosamine, sialic acid, N-acetylglucosamine Glucosamine, sulfoquinovose (6-deoxy-6-sulfo-D-glucopyranose) sucrose), ribose, arabinose, xylose, lyxose, sorbitol, mannitol , sucrose, lactose, maltose, trehalose, maltodextrin, raffinose It contains stachyose, glucuronic acid (glucuronide), and stachyose. L-form, 5-atom furanose-form, 6-atom cyclic pyranose-form, or acyclic, α-isomer (-OH of the anomeric carbon below the plane of the carbon atoms in the Haworth projection), or β-isomer (Haworth It can be the -OH of the anomeric carbon above the plane of the Worth projection. The term "ol" is used herein to refer to an oligosaccharide containing 3 to 6 sugar units.

[0068] "Pharmaceutically" or "pharmaceutically acceptable" means that a substance can be administered to an animal or human in an appropriate manner. "Inactive Ingredients" refers to molecules and compositions that do not cause adverse, allergic or other untoward reactions in the context of the intended use.

[0069] A "pharmaceutically acceptable solvate" or "solvate" is disclosed as a compound containing one or more solvent molecules. Examples of solvents that form pharmacologically acceptable solvates include water. , isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanol These include, but are not limited to, phenolamines.

[0070] Pharmaceutically acceptable auxiliary materials include any carrier, diluent, adjuvant or excipient, e.g. , preservatives, antioxidants, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, solvents, dispersion media, These include antibacterial agents, antifungal agents, isotonic agents, and absorption delaying agents. It is common practice to add these auxiliary materials to drug components that have the desired properties. It is reasonable to add auxiliary materials to the active pharmaceutical ingredient, unless the auxiliary materials are incompatible with the active pharmaceutical ingredient. It can be said that in order to achieve good results, active auxiliary materials are added to the drug components. Good too.

[0071] In the present invention, the term "pharmaceutical salt" refers to a salt derivative of the compound of the present invention. By suitable modification, the compounds of the present invention can be formed into the corresponding acid or alkali salts. Possible salts include the conventional non-toxic salts or quaternary ammonium salts, which are not intended to be limiting unless otherwise specified. The compound can be prepared by using a non-toxic inorganic or organic acid corresponding to the compound. Examples of the acid include hydrochloric acid, hydrobromic acid, sulfuric acid, aminosulfonic acid, phosphoric acid, and nitric acid, and the organic acid Acids include acetic acid, propionic acid, succinic acid, tartaric acid, citric acid, methanesulfonic acid, benzoic acid, Toluene sulfonic acid, glucuronic acid, glutamic acid, benzoic acid, salicylic acid, toluene sulfonate Acids that are used in pharmaceutically acceptable salts include carboxylic acids, oxalic acids, fumaric acids, and lactic acids. Other salts include trometamol, meglumine, and pyrroleethanol. ammonium salts such as ammonium salts, and sodium, potassium, calcium, zinc, magnesium, etc. Contains metal salts of

[0072] In the present invention, pharmaceutical salts are prepared by conventional chemical methods containing acidic or basic residues. Generally, these salts can be prepared from the parent compound by dissolving the salt in water, an organic solvent, or both. In a mixed solvent of the free acid or free base form of these compounds and a stoichiometric amount of an appropriate salt It can be obtained by reaction with a base or an acid. Ethers are generally used as non-aqueous reaction solvents. ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. For a list of salts, see Remington's Pharmaceutical Sciences iences”, 17th edition. Mack Publishing Company, Eas ton, PA, 1985, p. 1418, the disclosure of which is incorporated by reference. .

[0073] "Administering" or "Administration" ion) means any manner of transferring, delivering, introducing or transporting medicines or other drugs to a subject. Such modes include oral administration, topical contact, intravenous, intraperitoneal, intramuscular, lesional, nasal, and the like. Also contemplated by the present invention are intrathecal, intravenous, or intracavitary administration. Such devices may utilize active or passive transport. The delivery device may be a slow release or a fast release delivery device.

[0074] The novel conjugates disclosed herein utilize bridge linkers. Examples of bridge links and their synthesis are shown in Figures 1-34.

[0075] cross-linked body

[0076] Similar to the preparation of the cell-binding molecule-drug conjugates of the present invention, synthetic routes to obtain cross-linkers are described below. 1-20. The bridge linker has two components: a) a substituted acrylic or propyl group; It is a substituent that is an ol group and reacts with one or more thiols to form a thioether bond. and b) groups capable of reacting with a drug, including but not limited to: Disulfides, maleimides, haloacetyls, aldehydes, ketones, azides, amines, alkane Koxiamine, hydrazide, ethenesulfonyl, acyl halide (acid halide), Acrylic and / or anhydride groups, etc. Amide, ester, or thioester groups Substituted acrylic or propyl groups containing amine, alcohol, or thiol groups for forming bonds The synthesis of these bridge linkers and their application to antibody conjugation is shown in Figure 1. An example is given in ~20.

[0077] Preferably, the bridge linker is a compound of formula (I) or (II): [ka]

[0078] During the ceremony, [ka] represents a single bond, and when bonded to a single bond or a double bond, [ka] may be an enantiomeric or stereoisomeric bond; [ka] represents either a single bond, a double bond, or a triple bond; [ka] represents a single bond, both Lv1 and Lv2 are not H; [ka] represents a double bond, either Lv1 or Lv2 can be H, but at the same time It will not become H; [ka] represents a triple bond, Lv1 is absent and Lv2 may optionally be H;

[0079] Lv1 and Lv2 represent the same or different leaving groups that can be displaced by a thiol; The leaving group may include, but is not limited to, a halide (fluoride, chloride, bromide, and and iodide), methanesulfonyl (mesyl), toluenesulfonyl (tosyl), trifluoromethylsulfonyl (triflate), trifluoromethylsulfonyl Nitrophenoxyl, N-succinimidyloxyl (NHS), phenoxyl; Nitrophenoxyl; Pentafluorophenoxyl, Tetrafluorophenoxyl, Trifluorophenoxyl Fluorophenoxyl, difluorophenoxyl, monofluorophenoxyl, pentacene Chlorophenoxyl, 1H-imidazol-1-yl, chlorophenoxyl, dichlorophenoxyl Noxyl, trichlorophenoxyl, tetrachlorophenoxyl, N-(benzotriazo 2-ethyl-5-phenylisoxazolium-3'-sulfonyl, Phenyloxadiazole-sulfonyl (-sulfone-ODA), 2-ethyl-5-phenyl Nylisoxazolium-yl, phenyloxadiazol-yl (ODA), oxadiazol-yl or an intermediate produced by a condensation reagent for the Mitsunobu reaction. ru;

[0080] Y reacts with a drug or cytotoxic agent to form a disulfide, ether, ester, or thioate. esters, thioesters, peptides, hydrazones, carbamates, carbonates, amines (secondary , tertiary, or quaternary), imines, cycloheteroalkanes, heteroaromatics, alkyl oximes or a functional group capable of forming an amide bond; preferably, Y has the structure Has:

[0081] [ka] TIFF0007643734000031.tif97170

[0082] In the formula, X 1’ is F, Cl, Br, I, or Lv3; X 2’ are O, NH, N(R1 ), or CH2; R3 and R5 are each independently H, R1, aromatic, heteroaromatic Aromatic group, or one or more H atoms are -R1, -halogen, -OR1, -SR1, -NR substituted with -1R2, -NO2, -S(O)R1, -S(O)2R1, or -COOR1 Lv3 is an aromatic group; Lv4 is nitrophenol; N-hydroxysuccinimide (NHS ); Phenol; Dinitrophenol; Pentafluorophenol; Tetrafluorophenol difluorophenol; monofluorophenol; pentachlorophenol; Fluoride;Imidazole;Dichlorophenol;Tetrachlorophenol;1-Hydroxy Tosylate;Mesylate;2-Ethyl-5-phenylisoxazol azolium-3'-sulfonate, acid anhydrides formed with itself or with other acid anhydrides (e.g. For example, acetic anhydride, formic anhydride; or for peptide coupling reactions, or Mitsuno a leaving group selected from intermediates generated by condensation reagents for the BB reaction;

[0083] R1 is absent or C1-C8 alkyl; C2-C8 heteroalkyl, aralkyl Arylcycloalkyl, heterocycloalkyl; C3-C8 aryl, Ar-alkyl; Heterocycle, carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl , heteroaryl; or C2-C8 (carbon number 2 to 8) ester, ether, or amide or a peptide containing 1 to 8 amino acids; or a peptide having the structural formula (OCH2CH2) p Young is (OCH2CH(CH3)) p (p: an integer of 0 to about 1000) position, or a combination of the above groups;

[0084] Additionally, R1 is a chain of atoms selected from C, N, O, S, Si, and P, and is preferably or 0 to 500 atoms, and is covalently bonded to Y and L1. Atoms used to form R1 is alkylene, alkenylene, alkynylene, ether, polyoxyalkylene, esther esters, amines, imines, polyamines, hydrazines, hydrazones, amides, ureas, semicalcium Carbazide, carbazide, alkoxyamine, alkoxyamine, urethane, amino acid, pepto forming an amide, acyloxyamine, or hydroxamic acid, or a combination thereof; The bond may be any chemically relevant bond, such as

[0085] T is CH2, NH, NHNH, N(R3), N(R3)N(R 3’ ), O, S, C2- C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 Aryl, Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcyclo Alkyl, alkylcarbonyl, heteroaryl; peptides containing 1 to 4 amino acid units The amino acids are preferably aspartic acid, glutamic acid, arginine, hydroxybenzoate ... Styrene, lysine, serine, threonine, asparagine, glutamine, cysteine, selenium Cysteine, tyrosine, phenylalanine, glycine, proline, tryptophan, lanine; or one of the following structures:

[0086] [ka]

[0087] During the ceremony, [ka] is the linkage site;

[0088] X1, X2, X3, X4, X5, X6, X 1’ , X 2’ , and X 3’ are independent of each other. NH;NHNH;N(R3);N(R3)N(R 3’ );O;S;C1-C6 alkyl C2-C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C 3-C8 aryl, Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkoxy or 1 to 8 amino acids wherein R and R 3’ are each independently H; C1-C8 alkyl; C2-C8 hetero alkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl; Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcycloalkyl, a alkylcarbonyl, heteroaryl; C1-C8 esters, ethers or amides; or structural formula (OCH2CH2) p or (OCH2CH(CH3)) p (p;0~about 1 000) or a combination of the above;

[0089] m, m1, m2, m3, m4, and m5 each independently represent 1 to 10, preferably 1 is an integer between 0 and 4;

[0090] L1 and L2 are each independently the same or different and are selected from the group consisting of O, NH, S, NHNH, N( R3), N(R3)N(R 3’ ), formula (OCH2CH2) p OR3, or (OCH2 CH(CH3)) p OR3, or NH(CH2CH2O)p R3 or NH(C H2CH(CH3)O) p R3, or N[(CH2CH2O) p R3][(CH2C H2O) p’ R 3’ ], or (OCH2CH2) p COOR3 or CH2CH 2(OCH2CH2) p COOR3 polyethyleneoxy unit (wherein p and p' are each independently an integer of 1 to about 1000), or a combination thereof; C2-C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl C3-C8 aryl, Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroaromatic alkylcycloalkyl, alkylcarbonyl, heteroaryl; wherein R and R 3’ teeth each independently H; C1-C8 alkyl; C2-C8 heteroalkyl, alkylcyclo Aryl, Ar-alkyl, heterocycloalkyl; C3-C8 aryl, Ar-alkyl, heterocycle, Carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, hetero aryl; C1-C8 ester, ether, or amide; 1 to 8 amino acids; or Structural formula (OCH2CH2) p or (OCH2CH(CH3)) p (p;0~about 100 0 integer), or a combination of the above;

[0091] L1 or L2 may be a self-immolative or non-self-immolative component, a peptide unit, a hydrazone bond, a disulfide bond, or a It contains an amide bond, an ester bond, an oxime bond, an amide bond, or a thioether bond. The self-immolative unit may include, but is not limited to, 2-aminoimidazole- 5-methanol derivatives, heterocyclic PAB analogues, β-glucuronides, and o- or p-amino Electronically similar to the para-aminobenzylcarbamoyl (PAB) group of benzyl acetals, etc. Contains aromatic compounds;

[0092] Preferably, the self-immolative linker moiety has one of the following structures: [ka]

[0093] In the formula, ( * ) atoms labeled with an additional spacer or releasable linker unit, cell X is a binding site for damaging agents and / or cell adhesion molecules (CBAs); 1 , Y 1 , Z 2 , and Z 3 are independently NH, O or S; Z 1 are independently H, NHR1, OR1, SR1, COX1R1, where X1 and R1 are as defined above; v is 0 or 1. ru;U 1 are independently H, OH, C1-C6 alkyl, (OCH2CH2) n , F, Cl, Br, I, OR5, SR5, NR5R5', N=NR5, N=R5, NR5R5', NO 2, SOR5R5', SO2R5, SO3R5, OSO3R5, PR5R5', POR5 R5', PO2R5R5', OPO(OR5)(OR5'), or OCH2PO(OR5 )(OR5'), wherein R5 and R5' are each independently H, C1-C8 alkyl, alkyl; C2-C8 alkenyl, alkynyl, heteroalkyl, or amino acid; C3 -C8 aryl group, heterocycle, carbocycle, cycloalkyl, heterocycloalkyl, hetero an aralkyl group, an alkylcarbonyl group, or a glycoside group; or a medicinal cationic salt. be;

[0094] The non-self-immolative linker component is one of the following structures: [ka] TIFF0007643734000036.tif207164

[0095] In the formula, ( * ) atoms labeled with an additional spacer or releasable linker unit, cell X is a binding site for damaging agents and / or cell adhesion molecules (CBAs); 1 , Y 1 , U 1 , R5 The definitions of R5′ and R5′ are as described above; r is 0 to 100; m and n are each independently The numbers range from 0 to 6.

[0096] More preferably, L1 or L2 is composed of one or more linker components as shown below: May also be: [ka] TIFF0007643734000038.tif253170TIFF0007643734000039.tif253170, and L- or D-, natural or unnatural peptides containing 1 to 20 amino acids.

[0097] More preferably, L1 or L2 may be a releasable linker. The term refers to at least one bond that can be broken down under physiological conditions, e.g., pH labile. , acid labile, alkaline labile, oxidative labile, metabolic labile, biochemical labile, or a linker containing an enzyme-labile bond. Physiological conditions that cleave such a bond are The process does not necessarily involve biochemical or metabolic processes, but instead involves hydrolysis or is a displacement reaction, e.g., a reaction in an endosome where the pH is lower than the cytosolic pH, and / or disulfides of intracellular thiols (glutathione at mmol concentrations in malignant cells) It is understood that the reaction conditions may be a type of general chemical reaction condition such as a dopant exchange reaction.

[0098] Releasable linkers (L, L, or L2 Examples of ) include, but are not limited to: Not:-(CR5R6) m (Aa) r (CR7R8) n (OCH2CH2) t , -( CR5R6) m (CR7R8) n (Aa) r (OCH2CH2) t , -(Aa) r (CR 5R6) m (CR7R8) n (OCH2CH2) t , -(CR5R6) m (CR7R8) n (OCH2CH2) r (Aa) t , -(CR5R6) m (CR7=R8)(CR9R1 0) n (Aa) t (OCH2CH2) r , -(CR5R6) m (NR 11 CO)(Aa) t (CR9R 10 ) n (OCH2CH2) r , -(CR5R6) m (Aa) t (NR 11 CO)(CR9R 10 ) n (OCH2CH2)r 、-(CR5R6) m ( OCO)(Aa ) t (CR9R 10 ) n (OCH2CH2) r 、-(CR5R6) m (OCNR7)(A to) t (CR9R 10 ) n (OCH2CH2) r 、-(CR5R6) m (CO)(Aa) t (CR9R 10 ) n (OCH2CH2) r 、-(CR5R6) m (NR 11 CO)(A to) t (CR9R 10 ) n (OCH2CH2) r 、-(CR5R6) m ( OCO)(Aa ) t (CR9R 10 ) n (OCH2CH2) r 、-(CR5R6) m (OCNR7)(A to) t (CR9R 10 ) n (OCH2CH2) r 、-(CR5R6) m (CO)(Aa) t (CR9R 10 ) n (OCH2CH2) r 、-(CR5R6) m -フェニル-(CO) (Aa) t (CR7R8) n 、-(CR5R6) m -フリル-(CO)(Aa) t (CR 7R8) n 、-(CR5R6) m-oxazolyl-(CO)(Aa) t (CR7R8) n , -(CR5R6) m -thiazolyl-(CO)(Aa) t (CR7R8) n , -(CR5 R6) t -Thienyl-(CO)(CR7R8) n , -(CR5R6) t -Imidazolyl- (CO)(CR7R8) n , -(CR5R6) t -morpholino-(CO)(Aa) t (C R7R8) n , -(CR5R6) t -piperazino-(CO)(Aa) t (CR7R8) n , -(CR5R6) t -N-methylpiperazine-(CO)(Aa) t (CR7R8) n , -(CR5R) m -(Aa) t Phenyl-, -(CR5R6) m -(Aa) t Frills -, -(CR5R6) m -oxazolyl (Aa) t -, -(CR5R6) m -Thiazolyl (A a) t -, -(CR5R6) m -Thienyl (Aa) t -, -(CR5R6) m -Imidazo Lil (Aa) t -, -(CR5R6) m -Morpholino-(Aa) t , -(CR5R6) m -Piperazino-(Aa) t , -(CR5R6) m -N-methylpiperazino-(Aa) t , -K(CR5R6) m (Aa) r (CR7R8) n (OCH2CH2) t -K(CR5 R6) m (CR7R8) n (Aa) r (OCH2CH2) t -K(Aa) r (CR5R 6) m (CR7R8) n (OCH2CH2) t -K(CR5R6) m (CR7R8) n (OCH2CH2) r (Aa) t -K(CR5R6) m (CR7=R8)(CR9R1 0) n (Aa) t (OCH2CH2) r -K(CR5R6) m (NR 11 CO)(Aa ) t (CR9R 10 ) n (OCH2CH2) r -K(CR5R6) m (Aa) t (NR 11 CO)(CR9R 10 ) n (OCH2CH2) r -K(CR5R6) m (OCO) (Aa) t (CR9R 10 ) n (OCH2CH2) r -K(CR5R6) m (OCNR 7)(Aa) t (CR9R 10 ) n (OCH2CH2) r -K(CR5R6) m (CO (Aa)t (CR9R 10 ) n (OCH2CH2) r , -K(CR5R6) m (NR1 1CO)(Aa) t (CR9R 10 ) n (OCH2CH2) r , -K(CR5R6) m ( OCO)(Aa) t (CR9R 10 ) n (OCH2CH2) r , -K(CR5R6) m ( OCNR7)(Aa) t (CR9R 10 ) n (OCH2CH2) r , -K(CR5R6) m (CO)(Aa) t (CR9R 10 ) n (OCH2CH2) r , -K(CR5R6) m -phenyl-(CO)(Aa) t (CR7R8) n , -K(CR5R6) m -Frills-( CO)(Aa) t (CR7R8) n , -K(CR5R6) m -oxazolyl-(CO)( Aa) t (CR7R8) n , -K(CR5R6) m -thiazolyl-(CO)(Aa) t ( CR7R8) n , -K(CR5R6) t -Thienyl-(CO)(CR7R8) n , -K( CR5R6) t -Imidazolyl-(CO)(CR7R8) n , -K(CR5R6) t -Mo Rufolino-(CO)(Aa) t (CR7R8) n , -K(CR5R6) t -Piperazino- (CO)(Aa) t (CR7R8) n , -K(CR5R6) t -N-methylpiperazine- (CO)(Aa) t (CR7R8) n , -K(CR5R) m -(Aa) t Phenyl-, - K(CR5R6) m -(Aa) t Frill-, -K(CR5R6) m -oxazolyl (Aa ) t -, -K(CR5R6) m -Thiazolyl (Aa) t -, -K(CR5R6) m -Chie Nil(Aa) t -, -K(CR5R6) m -Imidazolyl (Aa) t -, -K(CR5R 6) m -Morpholino-(Aa) t , -K(CR5R6) m -Piperazino-(Aa) t ,- K(CR5R6) m -N-methylpiperazino-(Aa) t . (In the formula, the definitions of m, Aa, m, n, R3, R4, and R5 are the same as in formula (I). t and r are integers of 0 to 100; R6, R7, and R8 are each independently H , halogen, C1-C8 alkyl group, aryl group, alkenyl group, alkynyl group, ether esters, amines or amides (which contain one or more halogens, CN, NR1R2, CF 3. OR1, aryl group, heterocycle, S(O)R1, SO2R1, -CO2H, -SO3H , -OR1, -CO2R1, -CONR1, -PO2R1R2, -PO3H, or P(O ) optionally substituted with R1R2R3; K is selected from NR1, -SS- , -C(=O)-, -C(=O)NH-, -C(=O)O-, -C=NH-O-, -C= N-NH-, -C(=O)NH-NH-, O, S, Se, B, C3-C8 heterocycle or It is a peptide containing a heteroaromatic ring or 1 to 20 amino acids.

[0099] Furthermore, L1, L2, X1, X2, X3, X 1’ , X 2’ , and X 3’ are independent of each other. It does not have to exist.

[0100] In formula (I) or (II), the substituted acryl or propionyl group is a cell-binding The thiol of the agent can react with a pair of thiols, preferably a pair of thiols; Preferably, dithiothreitol (DTT), dithioerythritol (DTE), L-glucan glutathione (GSH), tris(2-carboxyethyl)phosphine (TCEP), 2- Mercaptoethylamine (β-MEA) or / and β-mercaptoethanol (β-ME The intrachain disulfide bonds of the cell-binding agent are reduced by a reducing agent such as 2-ME. It is a pair of sulfur atoms.

[0101] Examples of functional groups Y that allow for the attachment of drugs or cytotoxic agents include disulfides, thioethers, and the like. esters, thioesters, peptides, hydrazones, esters, carbamates, carbonates, Such functional groups include groups that allow for bonding via an alkoxide or amide bond. The groups include thiol, disulfide, amino, carboxyl, aldehyde, ketone, and methyl groups. imide, haloacetyl, hydrazine, alkoxyamino, and / or hydroxy. Examples include, but are not limited to:

[0102] Examples of functional groups Y that can react with the amine terminus of a drug / cytotoxic agent include, but are not limited to: However, N-hydroxysuccinimide ester, p-nitrophenyl ester, di Nitrophenyl ester, pentafluorophenyl ester, carboxylic acid chloride, or As a compound capable of reacting with the thiol terminal, including, but not limited to, pyridyl disulfide, nitropyridyl disulfide, maleimide , haloacetic acid, methylsulfonephenyloxadiazole (ODA), carboxylic acid chloride, and carboxylic acid anhydrides. Examples include, but are not limited to, amines, alkoxyamines, hydrazines, or Acyloxylamine can be used. Examples of amines that can react with the azide terminal include: It can be, but is not limited to, an alkyne.

[0103] In a preferred embodiment, R1, L1, or L2 each independently represents a 1 to 6 carbon atom. A straight chain alkyl group with a hydroxyl group, or the structural formula (OCH2CH2) p (p; an integer between 1 and 100 ) polyethyleneoxy units or 1 to 4 units of amino acids (L or D) or a combination of the above.

[0104] In a preferred embodiment, Lv1 and Lv2 are the same or each independently OG ;F;Cl;Br;I;Nitrophenol;N-hydroxysuccinimide (NHS); Phenol; Dinitrophenol; Pentafluorophenol; Tetrafluorophenol difluorophenol; monofluorophenol; pentachlorophenol; trifluorophenol Imidazole; Dichlorophenol; Tetrachlorophenol; 1-Hydroxy Benzotriazole; Tosylate; Mesylate; 2-Ethyl-5-phenylisoxazolidine It is formed by itself or with other acid anhydrides such as ammonium-3'-sulfonate, acetic anhydride, and formic anhydride. or for peptide coupling reactions or for Mitsunobu reactions The leaving group is selected from intermediates formed by condensation reagents for the purpose of For example, EDC (N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide), DCC (dicyclohexylcarbodiimide), N,N'-diisopropylcarbodiimide Dibromide (DIC), N-cyclohexyl-N'-(2-morpholinoethyl)carbodiimide so-p-toluenesulfonate (CMC, or CME-CDI), 1,1'-carbonyl Diimidazole (CDI), TBTU (O-(benzotriazol-1-yl)-N,N ,N',N'-tetramethyluronium tetrafluoroborate), N,N,N',N' -Tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluoro (Benzotriazol-1-yloxy)tris(dimethylamino)phosphate (HBTU), (amino)phosphonium hexafluorophosphate (BOP), (benzotriazole -1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyBO P), diethyl cyanophosphonate (DEPC), chloro-N,N,N',N'-tetrachloro Methylformamidinium hexafluorophosphate, 1-[bis(dimethylamino) methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide Hexafluorophosphate (HATU), 1-[(dimethylamino)(morpholino)methyl] ethylene]-1H-[1,2,3]triazolo[4,5-b]pyridin-1-ium-3- Oxidohexafluorophosphate (HDMA), 2-chloro-1,3-dimethylimide Dazolidinium hexafluorophosphate (CIP), chlorotripyrrolidinophosphonium PyCloP, Fluoro-N,N,N',N'-bis(trifluoromethyl) (tetramethylene)formamidinium hexafluorophosphate (BTFFH), N,N,N',N'-tetramethyl-S-(1-oxido-2-pyridyl)thiuronium Hexafluorophosphate, O-(2-oxo-1(2H)pyridyl)-N,N,N' ,N'-Tetramethylthiuronium tetrafluoroborate (TPTU), S-(1-O Oxido-2-pyridyl)-N,N,N',N'-tetramethylthiuronium tetrafluoride Roborate, O-[(ethoxycarbonyl)cyanomethyleneamino]-N,N,N',N '-Tetramethyluronium hexafluorophosphate (HOTU), (1-cyano- 2-Ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-cal Benzonium hexafluorophosphate (COMU), O-(benzotriazole-1-yl) (I)-N,N,N',N'-bis(tetramethylene)uronium hexafluorophosphine carboxylate (HBPyU), N-benzyl-N'-cyclohexylcarbodiimide (polymer binding with or without), dipyrrolidino(N-succinimidyloxy)-carbenium Hexafluorophosphate (HSPyU), chlorodipyrrolidinocarbenium hexafluorophosphate Phosphorophosphate (PyCIU), 2-chloro-1,3-dimethylimidazolinium tetrachloride Trifluoroborate (CIB), (benzotriazol-1-yloxy)dipiperidinyl O-(6-chlorobenzoxazole) Triazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoride Roborate (TCTU), Bromotris(dimethylamino)phosphonium hexafluoro Phosphate (BroP), Propylphosphonic Anhydride (PPACA, T3P (registered trademark) )), 2-morpholinoethyl isocyanide (MEI), N,N,N',N'-tetramethicone 1-O-(N-succinimidyl)uronium hexafluorophosphate (HSTU) , 2-bromo-1-ethyl-pyridinium tetrafluoroborate (BEP), O-[( Ethoxycarbonyl)cyanomethyleneamino]-N,N,N',N'-tetramethyluro Niobium tetrafluoroborate (TOTU), 4-(4,6-dimethoxy-1,3,5- Triazin-2-yl)-4-methylmorpholinium chloride (MMTM, DMTMM) , N,N,N',N'-tetramethyl-O-(N-succinimidyl)uronium tetra Fluoroborate (TSTU), O-(3,4-dihydro-4-oxo-1,2,3-benzyl) (benzotriazin-3-yl)-N,N,N',N'-tetramethyluronium tetrafluoro Oroborate (TDBTU), 1,1'-(azodicarbonyl)dipiperidine (ADD) , di-(4-chlorobenzyl) azodicarboxylate (DCAD), di-tert-butyl Diisopropyl azodicarboxylate (DBAD), diisopropyl azodicarboxylate (DIAD), or diethyl azodicarboxylate (DEAD). v1 and Lv2 may be acid anhydrides formed by themselves or with other C1-C8 acid anhydrides. stomach.

[0105] In a preferred embodiment, formula (I) or (II) has the following structure: [ka] TIFF0007643734000041.tif229121TIFF0007643734000042.tif245155TIFF0007643734000043.tif85150

[0106] Detailed examples of the synthesis of bridge linkers are shown in Figures 1-33. The bridging substituents, either acryloyl or disubstituted propanoyl, allow for the desired conjugation of the drug. The linker component can be condensed with a linker component that contains a functional group capable of reacting with the compound.

[0107] Cell-binding agent-drug conjugates

[0108] The conjugates of the present invention are represented by the following formulae (III), (IV), (V), (VI), (VII): The cell-binding agent-drug conjugate compound can be represented by (VIII) or (IX): [ka] TIFF0007643734000045.tif229134TIFF0007643734000046.tif62136

[0109] In the formula, n is 1 to 20. T has the same definition as in formula (I). Cb, Cb', C b''Cb''' may be the same or different and may be a cell binding agent / molecule or an immunotherapeutic protein , preferably an antibody or antibody fragment.

[0110] Inside the right brackets (square brackets) of formulas (III), (VII), (VIII), and (IX) is a linker-drug moiety that is conjugated to a thiol pair of a cell-binding agent / molecule. The esters are dithiothreitol (DTT), dithioerythritol (DTE), and dithiobutyl Amine (DTBA), L-glutathione (GSH), tris(2-carboxyethyl) phosphate Sulfonyl ether (TCEP), 2-mercaptoethylamine (β-MEA), or / and β-mercaptoethylamine The chains of the cell-binding agent are separated by a reducing agent selected from the group consisting of captoethanol (β-ME, 2-ME), A pair of sulfur atoms reduced from an inter-disulfide bond is preferred.

[0111] Drug, Drug', and Drug'' are the same or different cytotoxic agents or treatments. To promote the binding or stabilization of drugs, or immunotherapeutic proteins, or cell-binding agents R1 represents a functional molecule or a cell surface receptor binding ligand, and R1 represents a bridge linker of the present invention. and R1 is a C1-C8 alkane; Alkylene, alkenylene, alkynylene, aromatic ring, ether, polyoxyalkylene, Esters, amines, imines, polyamines, hydrazines, hydrazones, amides, ureas, semi- Carbazide, carbazide, alkoxyamine, urethane, amino acid, peptide, acylo xyamines, hydroxamic acids, disulfides, thioethers, thioesters, carbamates a carbonate, a heterocycle, a heteroalkyl, a heteroaromatic, or an alkoxide; Drug, Drug', and Drug'' can also include combinations of the above. Also, immunotherapeutic compounds, chemotherapeutic compounds, antibodies or antibody fragments, siRNA or It may also be a DNA molecule, or a cell surface-bound ligand.

[0112] [ka] represents a single bond or a double bond.

[0113] Inside the square brackets, a pair of sulfur atoms on the cell-binding molecule binds the cell-binding molecule. There are drugs that are conjugated with children.

[0114] m1, m 1’ , m 1’’ , m2, m 2’ , m 2’’ , m3, m4, m5, m 4’ , m5 ’ , m 4’’ , m 5’’ , m 4’’’ , m 5’’’ , m 4’’’’ , and m 5’’’’ , are each independently an integer of 1 to 10, preferably 1 to 4.

[0115] X1, X 1’ , X 1’’ , X 1’’’ , and X 2’’’’ are each independently NH; NHNH;N(R3);N(R3)N(R 3’ );O;S;C1-C6 alkyl;C2- C6 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C3-C8 Aryl, Ar-alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcycloa alkyl, alkylcarbonyl, heteroaryl; or 1 to 8 amino acids; 3 and R 3’ are each independently H; C1-C8 alkyl; C2-C8 heteroalkyl , alkylcycloalkyl, heterocycloalkyl; C3-C8 aryl, Ar-Al alkyl, heterocycle, carbocycle, cycloalkyl, heteroalkylcycloalkyl, alkylcarbocycle or a C1-C8 ester, ether, or amide; (OCH2CH2) p or (OCH2CH(CH3)) p (p; 0 to approximately 1000 (number of) polyethyleneoxy units, or a combination of the above, and further 1’ , X 1’’ , X 1’’’ , and X 2’’’’ may each independently be absent.

[0116] R1, R2, R 1’ , and R 1’’ are the same or different and are C1-C8 alkyl; C 2-C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C3-C 8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcyclo alkyl, alkylcarbonyl, heteroaryl; or C2-C8 ester, ether or amide; or the structural formula (OCH2CH2) p or (OCH2CH(CH3) ) p (p: an integer of 0 to about 1000), or a combination of the above groups. That is it.

[0117] L1, L 1’ , L 1’’ , L 1’’’ , L2, L 2’ , L 2’’ , and L 2’’’ ,teeth , have the same definition as L1 and L2 in formulae (I) and (II), and may be the same or different.

[0118] L1, L 1’ , L 1’’ , L 1’’’, L2, L 2’ , L 2’’ , and L 2’’’ ,teeth The linker may be composed of one or more linker components. The linker component may include, for example, 6-maleimide. Midocaproyl (MC), maleimidopropanoyl (MP), valine-citrulline (va l-cit or vc), alanine-phenylalanine (ala-phe or af), p- Aminobenzyloxycarbonyl (PAB), 4-thio-pentanoic acid ester (SPP) , 4-(N-maleimidomethyl)cyclohexane-1-carboxylic acid ester (MCC), (4-acetyl)aminobenzoic acid (SIAB), 4-thiobutyric acid ester (SPDB), 4-Thio-butyric acid ester (SPDB), 4-thio-2-hydroxysulfonyl-butyric acid ester ter(2-sulfo-SPDB), ethyleneoxy-C as one or more repeating units H2CH2O-(EO or PEO). Additional linker moieties are known in the art. and some are described herein.

[0119] Exemplary structures of the components of the linker include: [ka] TIFF0007643734000049.tif244170TIFF0007643734000050.tif244170

[0120] As described in more detail below, Drug, Drug', or Drug'' may refer to many The small molecule drug can be any of a wide variety of small molecule drugs, including, but not limited to: Tubulysins, calicheamicins, auristatins, maytansinoids, CC -1065 analogs, morpholinos, doxorubicins, taxanes, cryptophycins , amatoxins (amanitins), epothilones, geldanamycins (geldan amycins), duocarmycins, daunomycins, methotrexate, Vindesines, vincristines, and benzodiazepine dimers (e.g., pyrrolobenzodiazepines (PBDs), tomaymycin, indolinobenzodiazepines) azepines, imidazobenzothiadiazepines, or oxazolidinobenzodiazepines mers).

[0121] Generally, formulas (III), (IV), (V), (VI), (VII), (VIII), and and (IX) are produced from formulas (I) and (II), where "Drug" and "Cb" are , react with the formulas (I) and (II) individually or simultaneously. The acrylic or propiolic group reacts by addition reaction to form the compounds of formula (III), (IV), and (V) ), or (VI), wavelengths in the range of 190 to 390 nm, preferably 340 UV light of ∼380 nm, more preferably 365 nm, is preferably used to promote the reaction. Therefore, photochemical reactions are carried out in a temperature-controlled environment using quartz or Pyrex (registered trademark). The reaction was carried out in a continuous flow quartz tube (Trademark) flask or in an immersion well reactor containing a UV lamp. or in a Pyrex tube where UV irradiation is maximized (while allowing efficient cooling). This reduces thermal damage to cell-binding molecules. In forming (VII), (VIII), or (IX), two or more thiols may be of formula (I) and If the two or more substituted acrylic or propiolic groups of (II) are reacted with UV light, the UV light may optionally be Not necessary.

[0122] To synthesize the conjugate, first, the drug or cytotoxic molecule is dissolved in a chemical solvent or aqueous medium. with a linker of formula (I) or (II) to form formula (XVII) or (XVIII). The compound of formula (XVII) or (XVIII) can then be optionally isolated. , or immediately, simultaneously, or sequentially, DMA, DMF, at 25-38°C, pH 5-9. , ethanol, methanol, acetone, acetonitrile, THF, isopropanol, di Water-miscible organic solvents such as oxane, propylene glycol, or ethylenediol The disulfide bonds of cell-binding molecules were analyzed in aqueous media with or without the addition of 0-30% solvent. and reacting with a pair of free thiols generated through reduction of the thiol to form a compound of formula (III) or (IV) (V), or (VI) can be obtained, preferably with the aid of 365 nm UV light. or required to form formula (XVII), (XVIII), or (IX). For this purpose, UV light is not required.

[0123] Alternatively, with the aid of 365 nm UV light, a compound of formula (X), (XI), (XII), or with or without the aid of UV light to form a modified cell-binding molecule of formula (XIII). to form (XIV), (XV), or (XVI), Conjugates of (V), (VI), (VII), (VIII), and (IX) can also be first , water of pH 5 to 9 with or without the addition of 0 to 30% of water-miscible (miscible) organic solvent In an aqueous medium at 0 to 38°C, the linker of formula (I) or (II) is reacted with a pair of thiol groups on the cell-binding agent. The thiol pair can also be obtained by reacting it with a water-miscible (miscible) organic solvent. In aqueous media of pH 4-9 with or without the addition of 30% dithiothreitol (DT T), dithioerythritol (DTE), L-glutathione (GSH), tris(2-carboxylate) 2-mercaptoethylamine (β-MEA) and / or β-mercaptoethanol (β-ME, 2-ME) Therefore, it is preferable to reduce the pair of sulfur atoms from the interchain disulfide bonds of the cell-binding agent. Formula (III), (IV), (V), (VI), (VII), (VIII), and (IX) ) to obtain disulfides, maleimides, haloacetyls, azides, 1-ynes, ketones , aldehyde, alkoxyamino, triflate, carbonylimidazole, tosylate , mesylate, 2-ethyl-5-phenylisoxazolium-3'-sulfonate, or Nitrophenol, N-hydroxysuccinimide (NHS), phenol carboxylate esters; dinitrophenol, pentafluorophenol, tetrafluorophenol difluorophenol, monofluorophenol, pentachlorophenol, dichlorophenol chlorophenol, tetrachlorophenol, 1-hydroxybenzotriazole, anhydride, or formula (X) which may contain a hydrazide group or other acid ester derivatives , (XI), (XII), (XIII), (XIV), (XV), or (XVI) The functional group Y may be added simultaneously or sequentially with or without the addition of 0-30% of a water-soluble (miscible) organic solvent. Regardless of the drug / cytotoxicity, in an aqueous medium with a pH between 4 and 9.5, at 15 to 38°C, Reactivity of drugs / cytotoxic agents The groups react differently to the modified cell-binding molecule accordingly. The synthesis of cell-binding agent-drug conjugates linked via a hydroxyl bond can be performed by the synthesis of modified cell-binding agents. This is achieved by disulfide exchange between the disulfide bond in the hydroxyl group and a drug containing a free thiol group. The synthesis of cell-binding agent-drug conjugates linked via thioethers was reported by Maleimi et al. acetyl- or ethylsulfonyl-modified cell-binding agents and drugs containing free thiol groups The synthesis of conjugates bearing acid labile hydrazones is known in the art. This can be achieved by reacting the carbonyl group with the hydrazide residue in the linker by the method (e.g., P. Hamann et al., Hinman, LM, et al., Cancer Res. 53, 3336-334 , 1993; B. Laguzza et al., J. Med. Chem., 32; 548-555, 1959; P. Trail et al., Ca Cancer Res., 57; 100-105, 1997). The synthesis of conjugates with triazole bonds has been reported using click The 1-yne group in the drug and the azide residue in the linker are linked via chemistry (Huisgen cycloaddition). (Lutz, JF. et al, 2008, Adv. Drug Del. Rev. 60, 958-970; Sletten, EM et al 2011, Acc Chem. Research 44, 666-676). Oki Synthesis of cell-binding agent-drug conjugates linked via SIMS containing ketones or aldehydes This is achieved by reaction of a modified cell-binding agent with a drug containing an oxyamine group. To obtain cell-binding molecule-drug conjugates via ether bridges, thiol-containing drugs are Maleimide, haloacetyl, or ethylsulfonyl in aqueous buffer solution at pH 5.5-9.0 Substituted Formulae (X), (XI), (XII), (XIII), (XIV), and (XV) or (XVI) can be reacted with the modified cell-binding molecule conjugate of formula (XVI). To obtain a bridged conjugate, the thiol-containing drug is reacted with a compound of the formula ( X), (XI), (XII), (XIII), (XIV), (XV), or (XVI) Modified bridge linkers can be disulfide exchanged. Ether or thiol ether To obtain modified drugs with linkages, drugs with hydroxyl or thiol groups are In the presence of a cold base, for example, pH 8.0 to 9.5, halogens, especially carboxylic acid α-halides, Formula (X), (XI), (XII), (XIII), (XIV), (XV) having amides, or (XVI) to obtain an ester bridge. In the presence of a dehydrating agent such as EDC or DCC, drugs containing hydroxyl groups can be converted to carboxyl groups. and then condensing the cross-linker of formula (I) having a cross-linker group, and then condensing the cross-linker of interest with the drug-modified cross-linker. Conjugation with binding molecules can be carried out. To obtain conjugates via amide bond bridges, Drugs containing an amino group include those represented by formula (X), (XI), (XII), (XIII), (XIV), (XIII), (XIV), (XIV), (XIV), (XII ...II), (X XV) or (XVI) on the cell-binding molecule-linked complex, NHS, imidazole, nitrofen Carboxyl ester of phenol; N-hydroxysuccinimide (NHS); phenol dinitrophenol; pentafluorophenol; tetrafluorophenol; difluorophenol Monofluorophenol; Pentachlorophenol; Triflate; Imidazole;Dichlorophenol;Tetrachlorophenol;1-Hydroxybenzone Triazole; Tosylate; Mesylate; 2-Ethyl-5-phenylisoxazolium-3 It can be condensed with '-sulfonate.

[0124] The conjugates can be prepared by standard biochemical methods, e.g., Sephadex G25 or Sephac Gel filtration, adsorption chromatography, ion exchange, or permeation chromatography on a ryl S300 column In some cases, small molecules (e.g., When small molecule drugs (e.g., folic acid, melanocyte-stimulating hormone, EGF, etc.) are conjugated, chromatography, such as HPLC, medium pressure column chromatography, or ion exchange chromatography It can be purified by chromatography.

[0125] In a preferred embodiment, the compound of formula (III), (IV), (V), (VI), (VII), ( VIII) or (IX) has the following structure: [ka] TIFF0007643734000052.tif222164TIFF0007643734000053.tif217133TIFF0007643734 000054.tif234155TIFF0007643734000055.tif233160TIFF0007643734000056.tif35124

[0126] Modified cell-binding agents / molecules

[0127] The cell-binding agent modified by reaction with the conjugate of the present invention is preferably represented by formula (X), (XI) ), (XII), (XIII), (XIV), (XV), or (XVI): [ka] TIFF0007643734000058.tif155135

[0128] In the formula, R1, R 1’ , R 1’’ , R2, X1, X 1’ , X 1’’ , L1, L 1’ , L1 ’’ , L2, L 2’ , L 2’’ , [ka] Cb, m1, m 1’ , m 1’’ , m2, m 2’ , m 2’’ , m3, m4, m5, m 4’ , m 5’ , m 4’’ , and m 5’’ has the same definition as in formulas (III) to (IX).

[0129] During the ceremony, [ka] represents a single bond or a double bond.

[0130] In the formula, Y, Y', and Y'' have the same definitions as in formulas (I) and (II).

[0131] In a preferred embodiment, Y, Y', and Y'' are each independently a disulfide substituted groups, maleimides, haloacetyls, alkoxyamines, azides, ketones, aldehydes, hydrides Formed with aldehydes, alkynes, N-hydroxysuccinimide esters, or phenols carboxylic esters; dinitrophenol; pentafluorophenol; tetrafluorophenol difluorophenol; monofluorophenol; pentachlorophenol Triflate; Imidazole; Dichlorophenol; Tetrachlorophenol; 1 -Hydroxybenzotriazole; Tosylate; Mesylate; 2-Ethyl-5-phenyl isoxazolium-3'-sulfonate. Y, Y', and Y'' are each independently disulfides, thioethers, hydrazones, amides, alkoxides, carbamates independently reacting with cytotoxic agents via an ester, ether bond, or heteroaromatic ring. The modified cell-binding agent can be prepared by reacting the cell-binding agent with the compound represented by formula (III) above. and a linker of formula (I) or (II).

[0132] Substitution of a pair of free thiols on a cell-binding molecule, preferably an antibody, with a compound of formula (I) or (II). To achieve higher yields of the conjugation reaction with the acrylic or propiolic groups, the reaction mixture Similarly, the addition of a small proportion of an organic co-solvent to the compound of formula (I) in aqueous solution may be required. In order to maintain the solubility of II) to (IX), it may be necessary to add them to the solution after the reaction. To modify the cell-binding agent, first, a cross-linking reagent (cross-linking agent) of formula (I) or (II) is added. The compound is dissolved in a high concentration, for example, 1 to 500 mM, in a water-miscible polar organic solvent, for example, methanol. Different alcohols such as ethanol, propanol, acetone, acetonitrile, tetrahydrofuran tetrahydrofuran (THF), 1,4-dioxane, dimethylformamide (DMF), Dissolved in dimethylacetamide (DMA) or dimethyl sulfoxide (DMSO) On the other hand, it is possible to prepare a solution containing 1% hydroxybenzoate at a concentration of 1.0% in an aqueous buffer solution having a pH of 4 to 9.5, preferably 6 to 8.5. A cell-binding molecule such as an antibody dissolved at ~35 mg / ml is dissolved in 1 to 20 equivalents of TCEP or DT After reduction, DT was purified by SEC chromatography. TCEP can also be optionally removed by SEC chromatography. Alternatively, it can be retained in the reaction mixture for the next step reaction without purification. Furthermore, in order to achieve cross-linking of cell-binding molecules simultaneously with TCEP reduction, In addition, reduction of the antibody or other cell binding agent with TCEP results in the formation of a crosslinked antibody of formula (I) or (II). As described above, the compound represented by formula (X), (XI), (XII), or The formation of the modified cell-binding molecule (XIII) is assisted by UV light of 340-380 nm. The formation of the modified cell-binding molecule of formula (XIV), (XV), or (XVI) is then , optionally without the aid of ultraviolet light.

[0133] The aqueous solution for the modification of cell-binding agents has a pH between 4 and 9, preferably between 6.0 and 7.5. It can be buffered and contain non-nucleophilic buffer salts useful in these pH ranges. Agents include phosphate, acetate, triethanolamine HCl, HEPES, and MOP. S buffers, and further, for example, dextrin, sucrose, salts (e.g., NaCl, KC The solution containing the reduced cell-binding molecule may contain additional components such as a compound of formula (I). After adding the cross-linker (II) or (III), the reaction mixture is heated to 4°C to 45°C, preferably 15°C. Incubate at ambient temperature. The progress of the reaction can be monitored by the decrease in absorbance at 254 nm or By measuring the increase in absorbance at 280 nm or other suitable wavelength. After the reaction is complete, the modified cell-binding agent can be isolated using conventional methods, e.g., For example, this can be done by gel filtration chromatography or adsorption chromatography. .

[0134] The degree of modification can be determined by the UV spectrum emitted by nitropyridinethiones, dinitropyridines, and Lysine dithione, pyridine thione, carboxyamidopyridine dithione, and dicarboxy The development can be evaluated by measuring the absorbance of the thiamidopyridinedithione group. For conjugates without a chromophore group, the modification or conjugation reaction can be monitored by LC-MS, preferably UPL. Monitored by C-QTOF mass spectrometry or capillary electrophoresis-spectrometry (CEMS) The bridge linkers described herein may be any of the following with appropriate substituents: These have a variety of functional groups that can react with drugs, preferably cytotoxic drugs, such as amino or Modified cell-binding molecules with hydroxy substituents are prepared using N-hydroxysuccinimide (NHS) ) modified cell-binding molecules having thiol substituents that can react with drugs having esters; The molecule can react with drugs that have maleimide or haloacetyl groups. Modified cell-binding molecules with hydroxyl substituents (ketones or aldehydes) can be used in combination with hydrazides or aldehydes. Those skilled in the art will recognize that the available alkyloxyamines on the linker can react with drugs having alkyloxyamines. The linker to be used can be readily determined based on the known reactivity of the available functional groups.

[0135] Modified cytotoxic agents

[0136] The cytotoxic agent modified by reaction with the bridge linker of the present invention preferably has the formula (XVII) ) and (XVIII), wherein the drug "Drug" is a compound represented by the formula (I) and (II). A substituted thiol group that has reacted with a linker and is capable of reacting with a pair of thiols on a cell-binding agent. still have the thiol-reactive groups of the acrylic or propiolic groups: [ka]

[0137] During the ceremony, [ka] L1, L2, R1, T, m1, m2, m3, m4, m5, X1, Lv1, and Lv2 are Drug 1 has the same definition as in formula (I). Drug 1 has the same definition as in formula (II).

[0138] Formulas (XVII) and (XVIII) having a substituted acrylic or propiolic functional group via reaction of the drug with a linker of formula (I) and (II) to obtain a modified drug of formula (I): Drugs can be prepared. However, drugs containing thiols, or thioethers, thioesters or drugs that conjugate with cell-binding molecules via cross-linking agents via disulfide bonds. Therefore, Drug 1 first forms a thioether, thioester, or disulfide bond. It is preferable to connect the compound to one of the R1 components via a bond. Incorporating a substituted acryl group or a propionyl group into the R1-drug moiety gives the compound of formula (XVII) and and (XVIII) to form a crosslinker-modified drug.

[0139] In the synthesis example, to obtain the R1-Drug component having a thioether bond, The maleimide-containing drug was synthesized by the R1 segment of the linker bearing the maleimide substituent in aqueous buffer at neutral pH. and subsequently reacting the compound of formula (XVII) or (XV) having a thioether bond with III) to obtain the modified drug, by condensation with a substituted acrylic or propiolic group. To obtain an R1-Drug moiety having an ether bond, the hydroxy group can be The agent containing the halogen, tosylate, or mesylate is substituted in the presence of a mild base. and subsequently reacting the linker with the linker segment R1 of the formula ( To obtain the modified drug of formula XVII) or (XVIII), a substituted acrylic or proton group is added. The compound of formula (XVII) or (XVII) can be condensed with a pyrolysine group via an ester bond. To obtain the modified drug of II), the drug containing a hydroxy group is dehydrated with a dehydrating agent, such as EDC or In the presence of dicyclohexylcarbodiimide (DCC), a compound of formula (I) having a carboxyl group was reacted with ) can be condensed with a linker of the formula: To obtain the thiol-containing drug, a maleimide, vinylsulfonyl, or haloacetyl group is added. and subsequently reacting with a linker segment R1 having a thioether bond To obtain the modified drug of formula (XVII) or (XVIII), an acrylic or substituted The compound of formula (XVII) having an amide bond can be condensed with a component having a propiolic group. To obtain the modified drug of (XVIII), a drug having an amino group can be similarly reacted with a compound of formula (I) or can be condensed with the carboxyl group on the bridge linker of (II). Silica gel or alumina column chromatography, crystallization, preparative thin layer chromatography Purification can be performed by standard methods such as HPLC, ion exchange chromatography, or HPLC. This can be done.

[0140] In a preferred embodiment, formula (XVII) or (XVIII) has the following structure:

[0141] [ka] TIFF0007643734000064.tif230161TIFF0007643734000065.tif223146TIFF0007643734000066.tif139155

[0142] During the ceremony, [ka] Lv1 and Lv2 have the same definition as in formula (I); L1, L2, L3, L4, L5, L L6, L7, and L8 are the same or different and have the same definition as L1 in formula (I); Drug 1, Drug2, Drug3, Drug4, Drug5, Drug6, Drug7, and Dr Drug 8 has the same definition as Drug 1 in formula (II).

[0143] Cell-binding agents

[0144] The cell-binding molecule Cb constituting the conjugate and modified cell-binding molecule of the present invention can be used therapeutically. or other biologically modified residues of the cell population to be modified. It may be any molecule that is currently known or that will be discovered.

[0145] Cell binding agents include, but are not limited to, large molecular weight proteins, such as full-length antibodies (PPs). Reclonal or monoclonal), dimeric, multimeric, multispecific antibodies (e.g., bispecific single-chain antibodies; antibody fragments, e.g., Fab, Fab', F(ab')2, Fv [Parham, J. Immunol. 131, 2895-2902 (1983)], F Fragments obtained by ab expression libraries, anti-idiotypic (anti-Id) antibodies, CDRs, bispecific antibodies, trispecific antibodies, cancer cell antigens, viral antigens, microbial antigens, or The immunoglobulin is capable of recognizing and binding to a specific antigen or exhibiting a desired biological activity. epitope-binding fragments of any of the foregoing that immunospecifically bind to a protein produced in an immune system; Interferons (e.g., types I, II, III); peptides; lymphokines, e.g., IL-2, IL-3, IL-4, IL-5, IL-6, IL-10, GM-CSF, or Interferon gamma (IFN-γ); hormones, e.g., insulin, TRH (thyroid stimulator estrogen-releasing hormone), MSH (metastatic hormone), or androgen, estrogen steroid hormones such as steroids (e.g., steroid hormones) or melanocyte-stimulating hormone (MSH); growth factors and and colony-stimulating factors, e.g., epidermal growth factor (EFG), granulocyte-macrophage colony-stimulating factors stimulating factors (GM-CSF); transforming growth factors (TGF), e.g., TGF α, TGFβ; insulin and insulin-like growth factors (IGF-I, IGF-II) G -CSF, M-CSF, and GM-CSF [Burgess, Immunology T Today, 5, 155-158 (1984)]; Vaccine Growth Factor (VGF); Fibroblast Growth Factor (FGF) Follicular growth factors (FGFs); small molecular weight proteins, polypeptides, peptides, and peptide phosphatase inhibitors steroids, such as bombesin, gastrin, and gastrin-releasing peptide; platelet-derived Growth factors; interleukins and cytokines, e.g., interleukin-2 (IL-2 ), interleukin-6 (IL-6), leukemia inhibitory factor, granulocyte-macrophage colony -stimulating factor (GM-CSF); vitamins such as folic acid; apoproteins and glycoproteins, e.g. For example, transferrin [O'Keefe et al, J. Bio. Chem. 260 , 932-927 (1985)]; carbohydrate-binding proteins such as lectins or lipoproteins; vesicle nutrient transport molecules; and small molecule inhibitors, e.g., inhibition of prostate-specific membrane antigen (PSMA) drugs, small molecule tyrosine kinase inhibitors (TKIs), non-peptides, or other cell-binding molecules or materials, such as bioactive polymers (Dhar, et al., Proc. Natl. A cad.Sci.2008,105,17356-61), bioactive dendrimers (Le e,et al,Nat.Biotechnol.2005,23,1517-26;A lmutairi,et al;Proc.Natl.Acad.Sci.2009,1 06,685-90), nanoparticles (Liong, et al, ACS Nano, 200 8,19,1309-12;Medarova,et al,Nat.Med.2007 ,13,372-7;Javier,et al,Bioconjugate Chem .2008,19,1309-12), liposomes (Medinai,et al,Cu rr. Phar. Des. 2004, 10, 2981-9), viral capsid (Fle nniken,et al,Viruses Nanotechnol.2009,32 7,71-93).

[0146] Generally, if a suitable monoclonal antibody is available, it can be used to target the cell surface. Antibodies are preferred as binding molecules. Antibodies may be murine, human, humanized, chimeric, or derived from other species. So that's fine.

[0147] The antibodies used in the present invention can be produced using in vivo or in vitro production processes. The present invention also includes a method for preparing an anti-receptor peptide polyclonal antibody. is well known, for example, as shown in U.S. Pat. No. 4,493,795 (Nestor et al.). A typical method for preparing monoclonal antibodies is to generate antibodies from mice immunized with a specific antigen. This method involves fusing isolated mouse spleen cells with myeloma cells (Kohler ,G;Milstein,C.1975.Nature 256:495-497). Details For the correct procedure, see Antibodies-A Laboratory Manual. l, Harlow and Lane, eds., cold spring harbo Laboratory Press, New York (1988) The contents of this document are incorporated herein by reference. by immunizing a mouse, rat, hamster, or other mammal with the antigen of interest, Monoclonal antibodies can be obtained and used to target the antigen of interest, e.g., intact target cells. , antigens isolated from target cells, whole viruses, weakened whole viruses and viral proteins PEG6000 is used to fuse spleen cells with myeloma cells. The hybridomas obtained after the hybridization were screened using their sensitivity to HAT. Hybridomas producing monoclonal antibodies useful in the practice of the present invention are those that bind to specific targets. These antibodies are identified by their immunoreaction with target cell receptors or by their inhibition of receptor activity.

[0148] The monoclonal antibodies used in the present invention are those secreting antibodies with appropriate antigen specificity. Start a monoclonal hybridoma cell culture in a nutrient medium containing hybridoma cells. In the culture, the hybridoma cells release the antibody into the culture medium. The antibody-containing culture supernatant was collected and the conditions and time required for the antibody to be secreted into the culture medium were maintained. followed by well-known techniques, such as protein A affinity chromatography; anion exchange; Chromatography, cation exchange chromatography, hydrophobic interaction chromatography and molecular sieve chromatography (especially affinity chromatography using antigen-crosslinking protein A) chromatography and molecular sieve chromatography; centrifugation, sedimentation, or other methods The antibodies can be isolated by standard methods for protein purification.

[0149] Media useful for the preparation of these compositions are well known in the art and commercially available. and includes synthetic synthetic media. Exemplary synthetic media include Dulbecco's Minimum Essential Medium (DMEM) and MEM; Dulbecco et al., Virol. 8:396 (1959)), 4.5 g / L glucose, 0-20mM glutamine, 0-20% fetal bovine serum, ppm amounts of Some heavy metals (e.g., Cu, Mn, Fe, or Zn) or / and heavy metals added in salt form and antifoaming agents (e.g., polyoxyethylene-polyoxypropylene block copolymers) body).

[0150] In addition to cell fusion technology, we have also constructed cell lines for antibody production using the following methods: For example, direct transformation of B lymphocytes by oncogenic DNA can or oncogenic viruses, such as Epstein-Barr virus (EBV, human herpes simplex virus) Also known as pesvirus 4 (HHV-4) or Kaposi's sarcoma-associated virus Transfection with KSHV (see U.S. Patent No. 4,341,761 for details) ;4399121;4427783;4444887;4451570;4466917 ;4472500;4491632;4493890). Monoclonal antibodies are Based on known methods, anti-receptor peptides or peptides containing terminal carboxyl groups can be used. can be prepared (see Niman et al., Proc. Natl. Acad. Sc i. USA,80:4949-4953(1983);Geysen et al. Proc.Nat l. Acad. Sci. USA,82:178-182(1985); Lei et al. Bioc (See Hematology 34(20):6675-6688(1995)). The receptor polypeptide or polypeptide analog is a monoclonal antibody against the anti-receptor polypeptide. It may be used alone or conjugated to a cross-linked immunogenic carrier as an immunogen for preparing a steroid. It is possible.

[0151] To produce monoclonal antibodies as binding molecules of the present invention, other well-known production methods can be used. Among them, the method of producing fully human antibodies has attracted particular attention. Display technology uses affinity selection to select antibodies specific to a known antigen from a fully human antibody library. Phage display technology itself is widely used in the literature to obtain fully human antibodies that bind to specific antigens. , vector construction, and library screening are described in detail. Dente et al. Gene. 148(1):7-13 (1994); Little et al. Biotechnol Adv.12(3):539-55(1994);Clacks on et al. Nature 352:264-628(1991); Huse et al. Scien See ce 246:1275-1281 (1989).

[0152] Monoclonal antibodies derived from non-human species (e.g., mice) using hybridoma technology For cloned antibodies, humanization is performed to avoid human anti-mouse antibodies when administered to humans. Among these, the well-known method for antibody humanization is the method of humanizing the complementarity-determining region (CDR) of the antibody. For more information, see U.S. Patent Nos. 5,859,205 and 6,859,205. , No. 797, 492; Liu et al., Immunol Rev. 222:9-27 (2008 );Almagro et al., Front Biosci.1;13:1619-33(200 8);Lazar et al., Mol Immunol.44(8):1986-98(2007 );Li et al. Proc.Natl.Acad.Sci.USA.103(10):355 7-62 (2006), the disclosure of which is incorporated by reference. The transgenic mouse, U, possesses most of the human immunoglobulin light and heavy chains. It can be prepared by immunizing herons, monkeys, and other mammals with an antigen. Examples of such mice include Xenomouse (Abgenix, Inc.), HuM ab-Mouse(Medarex / BMS), VelociMouse(Regene For details, see U.S. Patent Nos. 6,596,541, 6,207,418, No. 6,150,584, No. 6,111,166, No. 6,075,181, No. 5,922,5 Nos. 45, 5,661,016, 5,545,806, 5,436,149 and 5, See US Pat. No. 569,825. In the course of human therapy, mouse antibody variable region genes and human antibody The immunogenicity of chimeric antibodies produced in the human body by integrating the human constant region genes is Much lower than mouse antibodies (Kipriyanov et al., Mol Biotech nol.26:39-60(2004);Houdebine,Curr Opin B Biotechnol. 13:625-9 (2002)). The disclosure of said document is incorporated by reference. Furthermore, site-directed mutagenesis of antibody variable regions can be used to improve the quality of antibody parental sequences. This can improve compatibility and specificity (Brannigan et al., Nat Rev M ol Cell Biol.3:964-70(2002);Adams et al., J.Imm Univ. Methods. 231:249-60 (1999)). By replacing the cytotoxicity of the cytotoxicity domain with the cytotoxicity of the immune effector cells, the cytotoxicity of the immune effector cells can be effectively promoted. Toxic effects can be enhanced.

[0153] Immunospecific antibodies against malignant cell antigens can be obtained commercially or by several conventional techniques. For example, they can be obtained by chemical synthesis or recombinant expression techniques. Nucleotide sequences encoding immunospecific antibodies to Other similar databases, commercial sources, published literature, or routine cloning and It can be obtained by sequencing.

[0154] Besides antibodies, polypeptides or proteins can also be used as binding molecules to target cell surfaces. Interact with the corresponding receptor or epitope by binding, blocking, attacking, or other means These peptides or proteins bind to epitopes or their corresponding receptors. They do not have to belong to the immunoglobulin family, as long as they can bind effectively. The polypeptides are also isolated by a technique similar to that of phage display antibodies (Sza et al., rdenings,J Recept Signal Transduct Res.2 003;23(4):307-49). Peptides obtained from a random peptide library The polypeptide or protein fragments are similar in application to antibodies and antibody fragments. The molecule binds to the antigen by connecting to some macromolecule or medium via a binding molecule. These macromolecules include, but are not limited to, albumin, polymers, , liposomes, nanoparticles, or dendrimers.

[0155] The conjugates of the present invention can be used to treat cancer, autoimmune diseases, and infectious diseases. Antibodies used for binding include, but are not limited to, 3F8 (anti-G D2 antibody), abagovomab (anti-CA-125 antibody), abciximab (anti-CD41 antibody ( Integrin α-IIb), adalimumab (anti-TNF-α antibody), adalimumab (anti-E pCAM antibody, CD326; afelimomab (anti-TNF-α); afutuzumab (anti-CD 20 antibody), Alacizumab pegol (anti-VEGFR2 antibody), A LD518 (anti-IL-6 antibody), alemtuzumab (also known as Campus, MabCampus, anti-CD52 antibody), altumomab (anti-CEA antibody), anatumomab (anti-tag-72 antibody) ), anrukinzumab (IMA-638, anti-IL-13 antibody), apolizumab (anti-HL A-DR antibody), arcitumomab (anti-CEA antibody), acelizumab (anti-L-selectin ( CD62L) antibody), Atlizumab (also known as Tocilizumab, Actemra, Actemra (anti-IL-6 receptor antibody), Atorolimumab (anti-albuminuria antibody) monkey factor antibody), bapineuzumab (anti-β-amyloid antibody), basiliximab (simlec anti-CD25 (IL-2 receptor α chain) antibody), bavituximab (anti-host Sphatidylserine antibody), Bectumomab (also known as LymphoScan, anti-CD2 2 antibody), belimumab (also known as BENLYSTA, LymphoStat-B, anti-BAFF antibody), Benraliz Benralizumab (anti-CD125 antibody), bertilimumab (anti-CCL11 (eosinophilic steroid) Scintimun (anti-CEA-related antigen antibody), besilesomab (also known as Scintimun, anti-CEA-related antigen antibody), Ibuprofen (also known as Avastin, an anti-VEGF antibody), biciromab (also known as FibriScint, an anti-fibroblast growth factor antibody) Blin II beta chain antibody), bivatuzumab (anti-CD44v6 antibody), blinatumomab (blinat umomab) (also known as BiTE, anti-CD19 antibody), brentuximab (c AC10, anti-CD30 TNFRSF8 antibody), Briakinumab (anti-I L-12, IL-23 antibody), canakinumab (also known as Ilaris, anti-IL-1 antibody), Cantuzumab (also known as C242, anti-CanAg antibody), Capromab, Katsuma Xomab (also known as removab, anti-EpCAM, anti-CD3 antibody), CC49 (anti-TAG-72 antibody), cedelizumab (anti-CD4 antibody), certolizumab pegol (other: C IMZIA, anti-TNF-α antibody), cetuximab (also known as Erbitux, IMC-C225 , anti-EGFR antibody), sitatuzumab (anti-EpCAM antibody), cixutumumab b) (anti-IGF-1 antibody), clenoliximab (anti-CD4 antibody), clivatuzumab (Cliva tuzumab) (anti-MUC1 antibody), Conatumumab (anti-TRAIL-R2 antibody) ), CR6261 (anti-influenza A hemagglutinin antibody), dacetuzumab mab) (anti-CD40 antibody), daclizumab (also known as Zenapax, anti-CD25C (IL-2 receptor Anti-CD38 (cyclic ADP-receptor) antibody), Daratumumab (anti-CD38 (cyclic ADP-receptor) antibody) RANKL antibody), denosumab (also known as Prolia, anti-RANKL antibody), Mab (anti-B-lymphoma cell antibody), Dorlimomab, Dorlixizumab, E Ecromeximab (anti-GD3 ganglioside antibody), eculizumab (also known as :Soliris, anti-C5 antibody), edovacomab (anti-endotoxin antibody), edrecolomab ( Other names: Panorex, MAb17-A1, anti-EpCAM antibody), efalizumab (other names: Rapti va, anti-LFA-1 (CD11a) antibody), efungumab (also known as Mycogra b, anti-Hsp90 antibody), elotuzumab (anti-SLAMF7 antibody), Elsilimomab (anti-IL-6 antibody), enlimomab pegol (anti-ICAM-1 ( CD54 antibody), epitumomab (anti-episialin antibody), epratuzumab (anti-CD22 antibody), Erlizumab (anti-ITGB2 (CD18) antibody) Ertumaxomab (also known as Rexomun, anti-HER2 / neu, CD3 antibody) ), etaracizumab (also known as Abegrin, anti-integrin αvβ3), exibirmab ( Anti-hepatitis B surface antigen antibody (HBs antibody), Fanolesomab (also known as Neu troSpec, anti-CD15 antibody), faralimomab antibody (anti-interferon receptor antibody), Farletuzumab (anti-folate receptor 1 antibody), Ferbiz Felvizumab (antibody against respiratory syncytial virus), Fezakinumab ( anti-IL-22 antibody), Figitumumab (anti-IGF-1 receptor antibody), Fontolizumab (anti-IFN-γ antibody), foravirumab ) (anti-rabies virus glycoprotein antibody), fresolimumab (anti-TGF -β antibody), Galiximab (anti-CD80 antibody), Gantene rumab (anti-β-amyloid antibody), gavilimomab (anti-CD147 (basic n) antibody), gemtuzumab (anti-CD33 antibody), girentuximab (anti Carbonic anhydrase 9 antibody), Glembatumumab (also known as CR011, anti-GPNM B antibody), golimumab (also known as Simponi, anti-TNF-α antibody), gomiliximab (Gomilix imab (anti-CD23C (IgE receptor) antibody), Ibalizumab (anti-C D4 antibody), Ibritumomab (anti-CD20 antibody), Igovomab b) (also known as Indimacis-125, anti-CA-125 antibody), imciromab (also known as Myoscint, an anti-cardiac myosin antibody), infliximab (also known as Remicade, an anti-TNF-α antibody), Intetumumab (anti-CD51 antibody), Inolimomab b) (anti-CD25 (IL-2 receptor α chain) antibody), inotuzumab (anti-CD 22 antibody), ipilimumab (anti-CD152 antibody), iratumumab (anti-CD 30 (TNFRSF8) antibody), Keliximab (anti-CD4 antibody), Ravez Zumab (also known as CEA-Cide, anti-CEA antibody), Lebrikizumab (anti-IL- 13 antibody), Lemalesomab (anti-NCA-90 (granulocyte antigen) antibody), Lerdelimumab (anti-TGFβ-2 antibody), Lexatumumab ) (anti-TRAIL-R2 antibody), Libivirumab (anti-hepatitis B surface antigen anti- body), Lintuzumab (anti-CD33 antibody), Lucatumumab (anti-CD40 antibody), lumiliximab (anti-CD23 (IgE receptor ) antibody), mapatumumab (anti-TRAIL-R1 antibody), maslimomab (anti T cell receptor antibody), Matuzumab (anti-EGFR antibody), Mepolizumab (another Name: Bosatria, anti-IL-5 antibody), Metelimumab (anti-TGFβ-1 antibody) ), Milatuzumab (anti-CD74 antibody), Minretumomab ) (anti-TAG-72 antibody), Mitumomab (also known as BEC-2, anti-ganglionic acid Osidoantibody-GD3), Morolimumab (anti-rhesus factor antibody), Mota Motavizumab (also known as Numax, an anti-RS virus antibody), muromonab b)-CD3 (also known as Orthoclone OKT3, anti-CD3 antibody), nacolomab (anti C242 antibody), Naptumomab (anti-5T4 antibody), Natalizumab (also known as Tysabri, an anti-integrin α4 antibody, Nebacumab (an anti-endotoxin antibody) ), Necitumumab (anti-EGFR antibody), Nerelimomab ( anti-TNF-α antibody), nimotuzumab (also known as Theracim, Theraloc, anti-EGFR antibody), Nofetumomab, ocrelizumab (anti-CD20 antibody), odulimomab ( Alias: Afolimomab, anti-LFA-1 (CD11a) antibody), ofatumumab (also known as Arzerr a, anti-CD20 antibody), olarratumab (anti-PDGF-Rα antibody), Omari Omalizumuba (also known as Xolair, an anti-IgE Fc antibody), oportuzumab (O portuzumab) (anti-EpCAM antibody), Oregovomab (also known as OvaRex, anti-C A-125 antibody), otelixizumab (anti-CD3 antibody), pagibaxima Pagibaximab (anti-LTA antibody), Palivizumab (also known as Synagis, Abbosynagis, anti- Respiratory syncytial virus antibody), panitumumab (also known as Vectibix, ABX-EGF, anti-EGFR antibody) ), Panobacumab (anti-Pseudomonas aeruginosa antibody), Pascolizumab (anti-IL-4 antibody), Pemtumomab (also known as Theragyn, anti-MUC1 antibody) , pertuzumab (also known as Omnitarg, 2C4, anti-HER2 / neu antibody), pexelizumab Pexelizumab (anti-C5 antibody), Pintumomab (anti-adenocarcinoma antigen antibody), Priliximab (anti-CD4 antibody), pritumumab (anti-vivimin chin antibody), PRO140 (anti-CCR5 antibody), racotumomab (also known as 1E10, anti-(N-glycolylneuraminic acid (NeuGc, NGNA)-ganglioside (GM3) antibody), Rafivirumab (anti-rabies virus glycoprotein antibody) ), Ramucirumab (anti-VEGFR2 antibody), Ranibizumab (also known as Lucen tis, anti-VEGF-A antibody), Raxibacumab (anti-anthrax toxin, protective antigen) antibody), Regavirumab (anti-CMV glycoprotein B antibody), Reslizumab Reslizumab (anti-IL-5 antibody), rilotumumab (anti-HGF antibody), Rituximab (also known as MabThera, Rituxanmab, anti-CD20 antibody), Robatumumab mumab (anti-IGF-1 receptor antibody), rontalizumab (anti-IFN-α Antibody), Rovelizumab (also known as LeukArrest, anti-CD11, CD18 antibody) ), Ruplizumab (also known as Antova, anti-CD154 (CD40L) antibody), Satumomab (anti-TAG-72 antibody), Sevirumab (anti-CMV antibody) antibody), sibrotuzumab (anti-FAP antibody), sifalimumab (anti-IFN -α antibody), siltuximab (anti-IL-6 antibody), siplizumab (anti-CD 2 antibody), (Smart) MI95 (anti-CD33 antibody), solanezumab ( anti-β-amyloid antibody), sonepcizumab (anti-sphingosine-1-lysine Anti-episialin antibody), Sontuzumab (anti-episialin antibody), Stamulumab ulumab (anti-myostatin antibody), sulesomab (also known as LeukoScan, (anti-N CA-90 (granulocyte antigen) antibody))), Tacatuzumab (anti-α-fetoplast) protein antibody), tadocizumab (anti-integrin αIIbβ3 antibody), Talizumab (anti-IgE antibody), tanezumab (anti-NGF antibody), tapritum Taplitumomab (anti-CD19 antibody), Tefibazumab (also known as Aur exis, anti-clumping factor A antibody), telimomab, tenatumomab omab) (anti-tenascin-C antibody), teneliximab (anti-CD40 antibody), Teplizumab (anti-CD3 antibody), TGN1412 (anti-CD28 antibody), Ticilimumab (also known as Tremelimumab, anti-CTLA-4 antibody), Tigatuzumab mab) (anti-TRAIL-R2 antibody), TNX-650 (anti-IL-13 antibody), Tocilizma Atlizumab (also known as Actemra, RoActemra, (anti-IL-6 receptor antibody), Toralizumab (T oralizumab) (anti-CD154 (CD40L) antibody), tositumomab (anti-CD20 antibody), Trastuzumab (also known as Herceptin, an anti-HER2 / neu antibody), tremelimumab limumab (anti-CTLA-4 antibody), Tucotuzumab celmoleukin ukin (anti-EpCAM antibody), tuvirumab (anti-hepatitis B antibody), Urutki Urtoxazumab (anti-Escherichia coli antibody), Ustekinumab (also known as St elara, anti-IL-12, IL-23 antibody), Vapaliximab (anti-AOC3 (VAP-1) antibody), vedolizumab, (anti-integrin α4β7 antibody ), veltuzumab (anti-CD20 antibody), bepalimomab (anti-AOC3 (V AP-1 antibody), visilizumab (also known as Nuvion, anti-CD3 antibody), vitaxin (anti-blood angiogenesis integrin avb3 antibody), Volociximab (anti-integrin α5β1), Votumumab (also known as HumaSPECT, anti-tumor antigen CTAA16.8 8 antibody), Zalutumumab (also known as HuMax-EGFr, (anti-EGFR antibody), Zanolimumab (also known as :HuMax-CD4, anti-CD4 antibody), Ziralimumab (anti-CD147 (basic immune globulin (antibody), zolimomab (anti-CD5 antibody), etanercept (registration (registered trademark "Enbrel"), Alefacept (registered trademark "Amevive"), Abatacept (registered trademark "Orencia"), rilonacept (Arcalyst), 14F7[ anti-IRP-2 (iron regulatory protein 2) antibody], 14G2a (black from Nat. Cancer Inst. Anti-ganglioside GD2 antibody for rheumatoid arthritis and solid tumors), J591 (Weill Cornell Medicine) Anti-PSMA antibodies for treating prostate cancer from the CAL School, ), 225.28S [melanoma Anti-HMW-MAA (High Molecular Weight Melanoma Associated Antigen) antibody for Sorin Radiofarmaci SRL (Milan, Italy)], COL-1 (from Nat. Cancer Inst. for colorectal cancer and gastric cancer) anti-CEACAM3 antibody, CGM1), CYT-356 (registered trademark "Oncoltad"), prostate cancer), HNK20 (Ora Vax Inc. for respiratory syncytial virus), ImmuRAIT (IMMU from NOMEDICS for non-Hodgkin's lymphoma), Lym-1 (anti-HLA-DR10 antibody, Pe Regrine Pharm for tumors), MAK-195F [from Abbott / Knoll for sepsis, toxin synthesis] Anti-TNF (tumor necrosis factor; TNFA, TNF-α; TNFSF2) antibodies for shock , MEDI-500 [also known as T10B9, an anti-CD3 antibody for graft-versus-host disease from MediImmune Inc.] Antibody, TRαβ (T cell receptor α / β), ], RING SCAN [from Neoprobe Corp. Anti-TAG72 (tumor-associated glycoprotein 72) antibodies for breast, colon, and colorectal cancer ], Avicidin (anti-EPCAM (epithelial cell adhesion molecule) antibody), anti-TACSTD1 ( Tumor-associated calcium signal transducer 1 (TAC) antibody, anti-GA733-2 (gastrointestinal tumor Related protein 2) antibodies, anti-EGP-2 (epithelial glycoprotein 2) antibodies; anti-KSA antibodies; K S1 / 4 antigen; M4S; tumor antigen 17-1A; from NeoRx Corp. for colon cancer, ovarian cancer, and prostate cancer CD326; LYMPHOCIDE (IMMUNOMEDICS) FOR CANCER AND NON-HODGKIN'S LYMPHOMA , NJ), Smart ID10 (Protein Design Labs), Oncolym (Techniclone Inc., C A), Allomune (BioTransplant, CA), anti-VEGF antibody (Genentech, CA); CEAcide (Immunomedics, NJ), IMC-1C11 (ImClone Systems, NJ) , as well as cetuximab (ImClone, NJ).

[0156] Other antibodies as cell binding molecules / ligands include, but are not limited to, the following antigens: Contains: aminopeptidase N (CD13), annexin A1, B7-H3 (CD276 , various cancers), CA125 (ovarian), CA15-3 (carcinoma), CA19-9 (carcinoma), L 6 (carcinoma), Lewis Y (carcinoma), Lewis X (carcinoma), α-fetoprotein (carcinoma), C A242 (colon rectum), placental alkaline phosphatase (carcinoma), prostate-specific antigen (prostate) ), prostatic acid phosphatase (prostate), epidermal growth factor (carcinoma), CD2 (Hodgkin's disease) , NHL lymphoma, multiple myeloma), CD3ε (T-cell lymphoma, lung cancer, breast cancer, gastric cancer, egg CD19 (B-cell malignancies), CD20 (non-Hodgkin's lymphoma), CD22 (leukemia, lymphoma, multiple myeloma, systemic lupus erythematosus), CD30 (Hodgkin's lymphoma), CD33 (leukemia, autoimmune diseases), CD38 (multiple myeloma) myeloma), CD40 (lymphoma, multiple myeloma, leukemia (CLL)), CD51 (metastatic melanoma, sarcoma), CD52 (leukemia), CD56 (small cell lung cancer, ovarian cancer, Merkel cell carcinoma) and liquid tumors, multiple myeloma), CD66e (cancer), CD70 (metastatic renal cell carcinoma and non-homologous tumors) CD74 (multiple myeloma), CD80 (lymphoma), CD98 (cancer) , mucin (carcinoma), CD221 (solid tumor), CD227 (breast cancer, ovarian cancer), CD262 (non-small cell lung cancer and other cancers), CD309 (ovarian cancer), CD326 (solid tumors), CEA CAM3 (colorectal cancer, gastric cancer), CEACAM5 (carcinoembryonic antigen; CEA, CD66e) (breast cancer, colorectal cancer, and lung cancer), DLL4 (Δ-like-4), EGFR (epithelial growth factor) receptors, various cancers), CTLA4 (melanoma), CXCR4 (CD184, hematopoietic tumors, solid tumors) solid tumors), endoglin (CD105, solid tumors), EPCAM (epithelial cell adhesion molecule, bladder bladder, head and neck, colon cancer, NHL prostate cancer, and ovarian cancer), ERBB2 (epidermal growth factor receptor Condition 2: lung cancer, breast cancer, prostate cancer), FCGR1 (autoimmune disease), FOLR (folate receptor , ovarian cancer), GD2 ganglioside (cancer), G-28G (cell surface antigen glycolipid, melanoma) , GD3 idiotype (cancer), heat shock protein (cancer), HER1 (lung cancer, gastric cancer) , HER2 (breast cancer, lung cancer, and ovarian cancer), HLA-DR10 (NHL), HLA-DRB ( NHL, B-cell leukemia), human chorionic gonadotropin (carcinoma), IGF1R (insulin -like growth factor-1 receptor, solid tumors, hematologic cancers), IL-2 receptor (interleukin-2 receptor condition, T-cell leukemia and lymphoma), IL-6R (interleukin 6 receptor, multiple bone myeloma, RA, Castleman's disease, IL6-dependent tumors), integrins (for various cancers) αVβ3, α5β1, α6β4, αIIβ3, α5β5, αVβ5), MAGE-1 (cancer MAGE-2 (carcinoma), MAGE-3 (carcinoma), MAGE-4 (carcinoma), anti-transcriptional Spherin receptor (carcinoma), p97 (melanoma), MS4A1 (transmembrane 4-domain family) Lee A member 1, non-Hodgkin's B-cell lymphoma, leukemia), MUC1 or MUC1-KL H (breast cancer, ovarian cancer, cervical cancer, bronchial and gastrointestinal cancer), MUC16 (CA125) (ovarian cancer) ), CEA (colon), gp100 (melanoma), MART1 (melanoma), MPG (melanoma) , MS4A1 (Transmembrane 4-domain family A member 1, small cell lung cancer, NHL), Creolin, neuro-oncogene product (carcinoma), P21 (carcinoma), anti-(N-glycolylneuraminidase) Paratope of amino acid (breast cancer, melanoma cancer), PLAP-like testicular alkaline phosphatase (ovarian cancer, testicular cancer), PSMA (prostate cancer), PSA (prostate), ROBO4, TAG72 (tumor Tumor-associated glycoprotein 72, leukemia (AML, gastric cancer, colorectal cancer, ovarian cancer), membrane of T cells Transmembrane protein (cancer), Tie (CD202b), TNFRSF10B (tumor necrosis factor receptor receptor superfamily member 10B, cancer), TNFRSF13B (tumor necrosis factor receptor Myeloma superfamily member 13B, multiple myeloma, NHL, other cancers, RA and SLE ), TPBG (trophoblast glycoprotein, renal cell carcinoma), TRAIL-R1 (TNF-related apoptosis) cis-ligand receptor 1, lymphoma, NHL, colorectal cancer, lung cancer), VCAM-1 (CD 106, melanoma), VEGF, VEGF-A, VEGF-2 (CD309) (various cancers) Other tumor-associated antigens recognized by antibodies have already been reported (Ge rber, et al, mAbs 1:3, 247-253 (2009); Novellino et al, cancer immunol immunothe r. 54 (3), 187-207 (2005) Franke et al, cancer biother radiopharm. 2000, 15,45 9-76).

[0157] The cell binding agent, more preferably an antibody, may bind to tumor cells, virus-infected cells, microbial-infected cells, Parasite-infected cells, autoimmune cells, activated cells, myeloid cells, activated T cells, B cells, or mesenchymal stem cells It can be any agent capable of combating lanocytes. More specifically, A binding agent is any agent that can target any one of the following antigens or receptors: It can be used as an agent / molecule: CD3, CD4, CD5, CD6, CD7, CD8, CD 9, CD10, CD11a, CD11b, CD11c, CD12w, CD14, CD15 , CD16, CDw17, CD18, CD19, CD20, CD21, CD22, CD2 3, CD24, CD25, CD26, CD27, CD28, CD29, CD30, CD3 1, CD32, CD33, CD34, CD35, CD36, CD37, CD38, CD3 9, CD40, CD41, CD42, CD43, CD44, CD45, CD46, CD4 7, CD48, CD49b, CD49c, CD51, CD52, CD53, CD54, C D55, CD56, CD58, CD59, CD61, CD62E, CD62L, CD62 P, CD63, CD66, CD68, CD69, CD70, CD72, CD74, CD7 9, CD79a, CD79b, CD80, CD81, CD82, CD83, CD86, C D87, CD88, CD89, CD90, CD91, CD95, CD96, CD98, C D100, CD103, CD105, CD106, CD109, CD117, CD120 , CD125, CD126, CD127, CD133, CD134, CD135, CD1 38, CD141, CD142, CD143, CD144, CD147, CD151, C D147, CD152, CD154, CD156, CD158, CD163, CD166 , CD168, CD174, CD180, CD184, CDw186, CD194, CD 195, CD200, CD200a, CD200b, CD209, CD221, CD22 7, CD235a, CD240, CD262, CD271, CD274, CD276(B 7-H3), CD303, CD304, CD309, CD326, 4-1BB, 5AC , 5T4 (trophoblast glycoprotein, TPBG, 5T4, Wnt activation inhibitor 1 or W AIF1), adenocarcinoma antigen, AGS-5, AGS-22M6, activin receptor-like kinase 1 , AFP, AKAP-4, ALK, α integrin, αvβ6, aminopeptidase N, Amyloid beta, androgen receptor, angiopoietin 2, angiopoietin 3, Nexin A1, anthrax toxin protective antigen, anti-transferrin receptor, AOC3 (VAP -1), B7-H3, Bacillus anthrax, BAFF (B-cell activating factor), B-lymphoma cells, b cr-abl, bombesin, BORIS, C5, C242 antigen, CA125 (carbohydrate antigen) Hara125, MUC16), CA-IX (or CAIX, carbonic anhydrase 9), CALLA, CanAg, canine IL31, carbonic anhydrase IX, cardiac myosin, CCL11 (CC mochi) -Chemokine 11), CCR4 (CC chemokine receptor type 4, CD194), CCR5 , CD3E (epsilon), CEA (carcinoembryonic antigen), CEACAM3, CEACAM5 (carcinoembryonic antigen), CFD (factor D), Ch4D5, cholecystokinin 2 (CCK2R) , CLDN18 (claudin-18), clumping factor A, CRIPTO, FCSF 1R (colony stimulating factor 1 receptor, CD115), CSF2 (colony stimulating factor 2, granules) macrophage colony-stimulating factor (GM-CSF), CTLA4 (cytotoxic T lymphocyte antigen 4 (CTLA4) lymphocyte-associated protein 4), CTAA16.88 tumor antigen, CXCR4 (CD184), C XC chemokine receptor type 4, cADP-ribose hydrolase, Cyclin B1, CY P1B1, cytomegalovirus, cytomegalovirus glycoprotein B, dabigatran , DLL4 (Delta-like ligand 4), DPP4 (Dipeptidyl peptidase 4), DR5 (Death receptor 5), Escherichia coli Shiga toxin type 2, ED-B, EGFL7 (protein 7-containing) EGF-like domain), EGFR, EGFRII, EGFRvIII, endoglin (CD 105), endothelin B receptor, endotoxin, EpCAM (epithelial cell adhesion molecule), EphA2, episialin, ERBB2 (epidermal growth factor receptor 2), ERBB3, ERG (TMPRSS2ETS fusion gene), E. coli, ETV6-AML, FAP (fibroblast Activator protein α), FCGR1, α-fetoprotein, fibrin II, β chain, fibronectin ectodomain B, FOLR (folate receptor), folate receptor α, folate hydrolase F-protein of respiratory syncytial virus (RSV), Frizzled receptor, fucoprotein, Fos-related antigen 1, F protein of respiratory syncytial virus (RSV), Glycyl GM1, GD2 ganglioside, G-28 (cell surface glycolipid antigen), GD3 idioside Type, GloboH, Glypican 3, N-glycolylneuraminic acid, GM3, GMCS F receptor α chain, growth differentiation factor 8, GP100, GPNMB (transmembrane protein NMB), GUCY2C (guanylate cyclase 2C, guanylate cyclase C (GC-C), intestinal guanylate cyclase Anilyl cyclase, guanylate cyclase-C receptor, heat-stable enterotoxin receptor hSTAR), heat shock protein, hemagglutinin, hepatitis B surface antigen, hepatitis B Virus, HER1 (human epidermal growth factor receptor 1), HER2, HER2 / neu, HE R3 (ERBB-3), IgG4, HGF / SF (stem cell growth factor / cell dispersion factor), H HGFR, HIV-1, histone complex, HLA-DA (human leukocyte antigen), HLA-D R10, HLA-DRB, HMWMAA, human chorionic gonadotropin, HNGF, human cytoplasmic reticulocyte colony-stimulating factor (HCR) Cell scattering factor receptor kinase, HPV E6 / E7, Hsp90, hTERT, ICAM- 1 (intercellular adhesion molecule 1), idiotype, IGF1R (IGF-1, insulin-like growth factor receptor 1 (IGF-1) factor 1 receptor), IGHE, IFN-γ, influenza hemagglutinin, IgE, IgE Fc region, IGHE, IL-1, IL-2 receptor (interleukin 2 receptor), IL -4, IL-5, IL-6, IL-6R (interleukin 6 receptor), IL-9, IL -10, L-12, IL-13, IL-17, IL-17A, IL-20, IL-22, IL-23, IL-31RA, ILGF2 (insulin-like growth factor 2), integrin ( α4, α IIIb β3, αvβ3, α4β7, α5β1, α6β4, α7β7, αIIβ 3, α5β5, αvβ5), interferon-γ-inducible protein, ITAGA2, ITG B2, KIR2D, LCK, Le, legumain, Lewis-Y antigen, LFA-1 (lymphocyte Function-associated antigen 1, CD11a), LHRH, LINGO-1, lipoteichoic acid, LIV1A , LMP2, LTA, MAD-CT-1, MAD-CT-2, MAGE-1, MAGE- 2, MAGE-3, MAGEA1, MAGEA3, MAGEA4, MART1, MCP- 1. MIF (macrophage migration inhibitory factor or glycosylation inhibitory factor (GIF)), MS 4A1 (transmembrane 4 domain subfamily A member 1), MSLN (mesothelin), M UC1 (mucin 1, cell surface associated (MUC1) or Polymorphic epithelial elial mucin(PEM)), MUC1-KLH, MUC16(CA125), MCP1 (monocyte chemotactic protein 1), MelanA / MART1, ML-IAP, MP G, MS4A1 (transmembrane 4 domain subfamily A), MYCN, myelin-associated glycoprotein Protein, myostatin, NA17, NARP-1, NCA-90 (granulocyte antigen), Ne ctin-4 (ASG-22ME), NGF, neuronal apoptosis-regulating proteinase 1, NOGO-A, Notch receptor, nucleolin, Neu oncogene product, NY-BR-1 , NY-ESO-1, OX-40, Oxidized low-density lipoprotein (OxLDL), OY-T ES1, P21, non-mutant p53, P97, Page4, PAP, anti-(N-glycolyl) neuraminic acid), paratopes of PAX3, PAX5, PCSK9, PDCD1 (PD-1 , programmed cell death protein 1, CD279), PDGF-Rα, (platelet-derived Growth factor receptor α), PDGFR-β, PDL-1, PLAC1, PLAP-like testicular alkaloids Phosphatase, platelet-derived growth factor receptor β, sodium phosphate cotransporter, PMEL 17, Polysialic acid, Proteinase 3 (PR1), Prostate cancer, PS (phosphatidylserine) Phosphorylphosphoric acid), prostate cancer cells, Pseudomonas aeruginosa, PSMA, PSA, PSCA, rabies virus glycoprotein Protein, RHD (Rh polypeptide 1 (RhPI), CD240), Rhesus factor (Rh esus factor), RANKL, PhoC, Ras mutant, RG55, ROBO 4, respiratory syncytial virus, RON, sarcoma metastasis breakpoint, SART3, sclerostin, SLAMF7 (SLAM family member 7), selectin P, SDC1 (syndecan 1), sLe(a), somatomedin C, SIP (sphingosine-1-phosphate), Somatostatin, sperm protein 17, SSX2, STEAP1 (six transmembrane domain of the prostate gland 1) epithelial antigen), STEAP2, STn, TAG-22 (tumor-associated glycoprotein 72), mackerel Ivin, T cell receptor, T cell transmembrane protein, TEM1 (tumor epithelial marker 1), T ENB2, tenascin C (TN-C), TGF-α, TGF-β (transforming Growth factor β), TGF-β1, TGF-β2 (transforming growth factor β2), T ie(CD202b), Tie2, TIM-1(CDX-014), TN, TNF, TN F-α, TNFRSF8, TNFRSF10B (tumor necrosis factor receptor superfamily member 10B), TNFRSF13B (tumor necrosis factor receptor superfamily member -13B), TPBG (trophoblast glycoprotein), TRAIL-R1 (tumor necrosis apoptosis) Death-inducing ligand receptor 1), TRAILR2 (death receptor 5 (DR5)), and major related Tumor-specific glycosylation of calcium signal transducer 2, MUC1, TWEA K receptor, TYRP1 (glycoprotein 75), TRP-2, tyrosinase, VCAM-1 (CD106), VEGF, VEGF-A, VEGF-2(CD309), VEGFR- 1, VEGFR2, or vimentin, WT1, XAGE1, or any insulin growth factor Cells expressing the epidermal growth factor receptor, or any epidermal growth factor receptor.

[0158] In another specific embodiment, the cell-binding ligand-drug conjugate of the present invention via the linker is used in the treatment of cancers, including but not limited to adrenal gland cancer Interstitial cancer, anal cancer, bladder cancer, brain tumors (adult, brainstem glioma, children, cerebellar astrocytoma, cerebral astrocytoma) Cell tumors, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal and pineal tumors, visual pathway and hypothalamic tumors glioma), breast cancer, carcinoid tumor, gastrointestinal, carcinoma of unknown primary origin, cervical carcinoma, colon carcinoma, uterine Endometrial cancer, esophageal cancer, extrahepatic bile duct cancer, Ewing family tumor (PNET), extracranial malignant embryonal carcinoma Cell tumors, eye cancer, intraocular melanoma, gallbladder cancer, gastric cancer (stomach), germ cell tumors, extragonadal, gestational trophoblastoma cancer, head and neck cancer, hypopharyngeal cancer, pancreatic islet cell carcinoma, kidney cancer (renal cell carcinoma), laryngeal carcinoma, leukemia (acute lymphoblastic, acute myeloid, chronic lymphocytic, chronic myeloid, hairy cell), lip and oral cavity cancer, Liver cancer, lung cancer (non-small cell, small cell), lymphoma (AIDS-related, central nervous system, skin T-cell, Hodgkin's disease, non-Hodgkin's disease, malignant mesothelioma, melanoma, Merkel cell carcinoma, metastatic of unknown primary Squamous neck cancer, multiple myeloma and other plasma cell neoplasms, mycosis fungoides, myelodysplastic syndrome, bone Myeloproliferative syndrome, nasopharyngeal carcinoma, neuroblastoma, oral cancer, pharyngeal cancer, osteosarcoma, ovarian cancer (epithelial, reproductive) cell tumors, low malignant potential tumors), pancreatic cancer (exocrine gland, islet cell carcinoma), paranasal sinus and Nasal cavity cancer, parathyroid cancer, penile cancer, pheochromocytoma cancer, pituitary cancer, plasmacytoma, prostate cancer, striated muscle Sarcoma, rectal cancer, renal cell carcinoma (kidney cancer), renal pelvis and ureter (transitional cell), salivary gland cancer, Sézary syndrome , skin cancer, skin cancer (cutaneous T-cell lymphoma, Kaposi's sarcoma, melanoma), small intestine cancer, soft tissue cancer tumor, stomach cancer, testicular cancer, thymoma (malignant), thyroid cancer, urethral cancer, uterine cancer (sarcoma), abnormalities in children Cancer, vaginal cancer, vulvar cancer, Wilms' tumor.

[0159] In another specific embodiment, the cross-linked cell-binding molecule-drug conjugate of the present invention is Depending on the composition and method, it can be used to treat or prevent autoimmune diseases. Immune disorders include, but are not limited to, autoimmune gastric achlorhydria, chronic active hepatitis, acute disseminated encephalomyelitis, acute hemorrhagic leukoencephalitis, Addison's disease, agammaglobulinemia, alopecia areata, Amyotrophic lateral sclerosis, ankylosing spondylitis, anti-GMB / TBM nephritis, antiphospholipid syndrome, Antisynthetase syndrome, arthritis, atopic allergy, atopic dermatitis, autoimmune Aplastic anemia, autoimmune cardiomyopathy, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune Inner ear disease, autoimmune lymphoproliferative syndrome, autoimmune peripheral nervous system disease, autoimmune pancreatic inflammation, multiple autoimmune endocrinopathy types I, II, and III, autoimmune progestational dermatitis , autoimmune thrombocytopenic purpura, autoimmune uveitis, Barlow's disease / Barlow's concentric Sclerosis, Behçet's disease, Berger's disease, Bickerstaff's encephalitis, Blau syndrome group, bullous pemphigoid, Castleman's disease, Chagas disease, chronic fatigue immune dysfunction syndrome , chronic inflammatory demyelinating polyneuropathy, chronic recurrent multifocal osteomyelitis, chronic Lyme disease, chronic obstructive pulmonary disease Pulmonary disease, allergic granulomatous vasculitis, cicatricial pemphigoid, celiac disease, Cogan's syndrome group, cold agglutinin disease, complement component C2 deficiency, cephalic arteritis, crest syndrome, Crohn's disease (particularly inflammatory bowel disease), Cushing's syndrome, cutaneous leukocytoclastic vasculitis, malignant atrophic papulosis, painful steatosis, dermatitis herpetiformis, dermatomyositis, type 1 diabetes, diffuse cutaneous scleroderma, myocardial infarction, Discoid lupus erythematosus, eczema, endometriosis, enthesitis-associated arthritis, eosinophilic fasciitis, acquired inflammatory bowel disease Cutis bullosa, erythema nodosum, idiopathic mixed cryoglobulinemia, Evans syndrome, progressive ossification fibrodysplasia, fibromyalgia, fibromyositis, fibrosing alveolitis, gastritis, gastrointestinal pemphigoid, Giant cell arteritis, glomerulonephritis, Goodpasture's syndrome, Graves' disease, Guillain-Barré syndrome syndrome, Hashimoto's encephalopathy, Hashimoto's thyroiditis, hemolytic anemia, allergic purpura, herpes gestationis, suppurative Hidradenitis, Hughes syndrome (antiphospholipid syndrome), hypogammaglobulinemia, idiopathic inflammation Demyelinating diseases, idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura (autoimmune thrombocytopenic purpura) IgA nephropathy (Berger's disease), inclusion body myositis, inflammatory demyelinating polyneuropathy, interstitial Interstitial cystitis, irritable bowel syndrome, juvenile idiopathic arthritis, juvenile rheumatoid arthritis, mucocutaneous phosphorus Pain node syndrome, Lambert-Eaton myasthenic syndrome, leukocytoclastic vasculitis, lichen planus, sclerosis Lichen vegetans, Linear IgA Disease (LAD), Lou Gehrig's Disease (Amyotrophic Lateral Sclerosis), Wolff-Wolf Acne-like hepatitis, lupus erythematosus, Blau syndrome, Meniere's disease, microscopic polyangiitis, Miller-F Fisher syndrome, mixed connective tissue disease, scleroderma, Mucha-Jakob disease, Muckle-Wells syndrome syndrome, multiple myeloma, multiple sclerosis, myasthenia gravis, myositis, narcolepsy, optic nerve spinal cord injury Meningitis (Devic's disease), neuromuscular, ocular cicatricial pemphigoid, opsoclonus-myoclonus syndrome, Odo thyroiditis, relapsing rheumatoid arthritis, Panda syndrome (childhood autoimmune disease with complicated streptococcal infections) Autoimmune neuropsychiatric disorders, tumor-associated cerebellar degeneration, paroxysmal nocturnal hemoglobinuria, Parry-Romberg syndrome syndrome, Parsonage-Georgia syndrome, pars planus inflammation, pemphigus, pemphigus vulgaris, pernicious anemia , perivenous encephalomyelitis, POEMS syndrome, polyarteritis nodosa, polymyalgia rheumatica, poly Myositis, primary biliary cirrhosis, primary sclerosing cholangitis, progressive inflammatory neuropathy, psoriasis, ectopic arthritis, pyoderma gangrenosum, pure red cell aplastic anemia, Rasmussen's encephalitis, Raynaud's disease, Polychondritis reticulata, Reiter's syndrome, restless legs syndrome, retroperitoneal fibrosis, rheumatoid arthritis , rheumatic fever, sarcoidosis, schizophrenia, Schmidt syndrome, Schnitzler syndrome group, scleritis, scleroderma, Sjogren's syndrome, spondyloarthropathy, sticky blood syndrome, Still's disease, Stiff-man syndrome, subacute bacterial endocarditis, Susac syndrome, acute febrile neutrophilic epithelium Skin disease, Sydenham's chorea, sympathetic ophthalmia, Takayasu's arteritis, temporal arteritis (giant cell arteritis), Saunders-Hunt syndrome, transverse myelitis, ulcerative colitis (idiopathic inflammatory bowel disease), undifferentiated connective tissue Wegener's disease, undifferentiated spondyloarthropathy, vasculitis, vitiligo, Wegener's granulomatosis, Wilson's syndrome, Includes Stocott-Aldrich syndrome.

[0160] In another specific embodiment, the crosslinks of the present invention for the treatment or prevention of autoimmune diseases Linking molecules used for interbody conjugation include, but are not limited to, anti-elastic Antibody; Anti-epithelial cell antibody; Anti-basement membrane type IV collagen protein antibody; Anti-nuclear antibody Anti-double-stranded DNA antibody, anti-single-stranded DNA antibody, anti-cardiolipin antibody IgM, IgG; Anti-celiac antibodies; anti-phospholipid antibodies IgK, IgG; anti-SM antibodies; anti-mitochondrial antibodies ;Thyroid antibody; Microgranular antibody, T cell antibody; Thyroglobulin antibody, anti-scleroderma-70 antibody (A antiSCL-70; anti-Jo antibody (Anti-Jo), anti-U1RNP antibody (Anti -U1RNP);anti-La / SSB antibody;anti-SSA antibody;anti-SSB antibody;anti-parietal cell antibody;anti- Histone antibody; anti-RNP antibody; C-ANCA; P-ANCA; anti-centromere antibody; anti-F Antibody, anti-GBM antibody, anti-ganglioside antibody; anti-desmosomal glycoprotein 3 antibody Antibody (anti-Desmogein3); anti-p62 antibody; anti-sp100 antibody; anti-mitochondrial antibody Chondria (M2) antibody; rheumatoid factor antibody; anti-MCV antibody; anti-topoisomerase antibody; These include antineutrophil cytoplasmic (cANCA) antibodies.

[0161] In some preferred embodiments, the binding molecules used in the conjugates of the invention are By binding to receptors or receptor complexes expressed by disease-associated activated lymphocytes. The receptor or receptor complex can be, for example, a member of the immunoglobulin gene superfamily. Members of the CD11 receptor (e.g., CD2, CD3, CD4, CD8, CD19, CD20, CD 22, CD28, CD30, CD33, CD37, CD38, CD56, CD70, CD 79, CD90, CD125, CD147, CD152 / CTLA-4, PD-1, or ICOS), TNF receptor superfamily (e.g., CD27, CD40, CD95 / Fas, CD134 / OX40, CD137 / 4-1BB, INF-R1, TNFR- 2, RANK, TACI, BCMA, osteoprotegerin, Apo2 / TRAIL-R 1, TRAIL-R2, TRAIL-R3, TRAIL-R4, and APO-3), Tegrin, cytokine receptors, chemokine receptors, major histocompatibility proteins, lectins Examples of such proteins include complement regulatory proteins (type C, type S, or type I).

[0162] In another specific embodiment, the antibody is immunospecific for a viral or bacterial antigen. A useful conjugate is a humanized or human monoclonal antibody. The term "viral antigen" includes any viral peptide, polypeptide, or antigen capable of eliciting an immune response. glycoproteins (e.g., HIV gp120, HIV nef, RSV F glycoprotein, Influenza virus neuraminidase, influenza virus hemagglutinin, HTL Vtax, herpes simplex virus glycoproteins (e.g., gB, gC, gD, and gE), and hepatitis B surface antigen). The term "bacterial antigen" includes any microbial peptide, polypeptide, or antigen capable of eliciting an immune response. Protein, sugar, polysaccharide, or lipid molecules (e.g., bacteria, fungi, pathogenic protozoa, yeast, These include, but are not limited to, peptides (e.g., LPS and 5 / 8). Type I antibodies useful for treating RVS or bacterial infections include palivizumab (used to treat RVS infections). Humanized anti-respiratory syncytial virus monoclonal antibody), PRO542 (HIV infection) CD4 fusion antibody used to treat hepatitis B virus), Ostavir (a hepatitis B virus treatment) antibody), PROTVIR (humanized IgG1 antibody used to treat cytomegalovirus antibodies), and anti-LPS antibodies.

[0163] The cell-binding molecule-drug conjugates of the present invention via a cross-linker are useful for treating infectious diseases. The infectious diseases include Acinetobacter infection, actinomycosis, African sleeping sickness ( African trypanosomiasis), AIDS (acquired immune deficiency syndrome), amebiasis, anaplasia Zuma, Anthrax, Bacillus anthracis, Bacillus tuberculosis infection, Argentine hemorrhagic fever, Ascariasis, Aspergillosis, Ast Rovirus infection, babesiosis, Bacillus cereus infection, bacterial pneumonia, bacterial vaginosis, bacteriologic Ides infection, Balantidiosis, Bailey's roundworm infection, BK virus infection, black sand hair , Blastocys hominis infection, Blastomyces, Bolivian hemorrhagic fever, Borrelia infection, Bo Turinism (and infant botulism), Brazilian hemorrhagic fever, brucellosis, Burkholderia infection, Buruli ulcer, Calicivirus (Norovirus, Sapovirus), Campylobacter pylori infection Lactobacillus infection, Candida infection (candidiasis, thrush), cat scratch disease, Cellulitis, Chagas disease (American trypanosomiasis), chancroid, chickenpox, plaque, plaque pneumonia Plasmodium infection, cholera, pigmented fungus, liver fluke disease, Clostridium difficile infection, Schizoidomycosis, Colorado tick fever, colds (acute viral nasopharyngitis, acute rhinitis), black flu Zfeldt-Jakob disease, Crimean-Congo hemorrhagic fever, Cryptococcus, Cryptosporidium Umbilical cord disease, skin larval migration, Cyclospora infection, cysticercosis, cytomegalovirus infection, dengue Fever, diamoebiasis, diphtheria, diphyllobothriasis, dracunculiasis, Ebola hemorrhagic fever, hydatid disease, Urrichiasis, pinworms (pinworm infection), enterococcal infection, enterovirus infection, typhus Erythema infectiosum (fifth disease), acute exanthema in children, trematode fluke disease, thrombocytopenia, fatal familial insomnia , filariasis, food poisoning caused by Clostridium perfringens, non-parasitic amoebic infections, Fusobacterium infection, gas gangrene (clostridial myonecrosis), diotrichosis, gel Stomann-Straussler-Scheinker syndrome, Giardiasis, glanders, gnathostomiasis, Gonorrhea, granuloma groin (donovanosis), group A streptococcus infection, group B streptococcus infection, influenza Hand, foot and mouth disease (HFMD), Hantavirus pulmonary syndrome, Helicobacter pylori Lori infection, hemolytic uremic syndrome, hemorrhagic fever with renal syndrome, hepatitis A, hepatitis B, hepatitis C, hepatitis D Hepatitis, Hepatitis E, Herpes simplex, Histoplasmosis, Hookworm infection, Human Balkan Virus infection Dye, human ehrlichiosis Evans, human granulocytic anaplasmosis, human metapneumovirus human monocytic aerosol infection, human papillomavirus infection, human parainfluenza Virus infection, microtaeniasis, influenza, isosporosis, Kawasaki disease, mononucleosis , Kim's disease, kuru, Lassa fever, Legionnaires' disease, Legionnaires' disease leishmaniasis, leprosy, leptospirosis, listeriosis, leprosy Lyme disease (Lyme borrelia), lymphatic filariasis (elephantiasis), lymphocytic choriomeningitis, ria, Marburg hemorrhagic fever, measles, melioidosis (Whitmore's disease), meningitis, meningococcal disease , metagonism, microsporidiosis, molluscum contagiosum, mumps, typhus (endemic typhus), mycoplasma pneumonia, mycetoma, myiasis, neonatal conjunctivitis (ophthalmia neonatorum), Zfeldt-Jakob disease (vCJD, nvCJD), nocardiosis, onchocerciasis (blindness) filariasis), paracoccidioidomycosis (South American blastomycosis), paragonimiasis, pasture disease Head lice, head lice, body lice, pubic lice (Pubic lice, Crabb ice), Pelvic inflammatory disease, Whooping cough ugh), plague, pneumococcal infection, Pneumocystis carinii pneumonia, pneumonia, polio, Prevotella infection, PA ME, progressive multifocal leukoencephalopathy, psittacosis, Q fever, rabies, rat bite fever, respiratory syncytial Viral infections, rhinovirus infections, rickettsial infections, rickettsia, Rift Valley fever, Rocky Mountain spotted fever, rotavirus infection, rubella, salmonellosis, SARS (severe acute respiratory syndrome), Organ syndrome), scabies, schistosomiasis, sepsis, diarrhea (dysentery), shingles (Herpes zoster) ster), smallpox, sporotrichum, staphylococcal food poisoning, staphylococcal infection, nematodes, syphilis , Taeniasis, Tetanus (Trismus), Barber's itch, Tinea Hands , pityriasis nigricans, tinea pedis, tinea unguium, tinea versicolor, toxocariasis (ocular larva migrans), Toxocara visceral larva migrans, toxoplasmosis, trichinosis, trichomoniasis, cryptobiosis (Whipworm infection), pulmonary tuberculosis, tularemia, urea-degrading Mycoplasma infection, Venezuelan equine Encephalitis, Venezuelan hemorrhagic fever, viral pneumonia, West Nile fever, rhizobacterium balanitis, Mycobacterium pseudotuberculosis Infectious diseases, including but not limited to yersiniosis, yellow fever, and zygomycosis.

[0164] A cell-binding molecule, preferably an antibody against the pathogenic strain described herein, Strains include Acinetobacter baumannii, Actiomyces israelii, and Actinomyces Actinomyces odontolyticus, Rhopionibacterium propionicus, Trypanosoma brucei, HIV (human immunodeficiency virus) Virus), Entamoeba histolytica, Anaplasma spp., Bacillus anthracis, Methylschaemolyticum ( Arcanobacterium haemolyticum), Junin virus, roundworms , Aspergillus, Astroviridae, Babesia, Bacillus cereus, Multiple Bacteria, Bacteroides, Colonic Pouch Ciliates, Baileyi Roundworm, Nematode, BK Virus , Piedraiahortae, Blastocystis hortae Minis, dermatitis spp., Macupo virus, Borrelia, Clostridium botulinum, Sabia, Brucella Burkholderia genus, usually Burkholderia cepacia and other Burkholderia species, Mycobacteria M. ulcerans, Caliciviridae family, Campylobacter, usually Candida albicans and other Candida species, Bartonella henselae henselae), group A streptococci and staphylococci, Trypanosoma cruzi, and soft papules Bacillus casei, varicella-zoster virus (VZV), Chlamydia trachomatis, Chlamydia nuclei -monier, Vibrio cholerae, Fonseca pedrosoi, Clonorchiasis, Clostridium diff Icyle, Coccidioides immitis, Coccidioides posadasii, Colorado tick fever Viruses, rhinoviruses, coronaviruses, Creutzfeldt-Jakob disease, prions, Crimean-Congo hemorrhagic fever virus, Cryptococcus neoformans, Cryptosporidium Rhydium spp., Cat hookworm, Co-parasites, Cyclospora, Taenia solium, Cytomegalovirus, Den DEN-1, DEN-2, DEN-3, and DEN-4 viruses - Flaviviruses , Dictyostelium, Corynebacterium diphtheriae, Diphyllobothrix, Dracunculia spp. nculusmedinensis), Ebola virus, Echinococcus, Ehrlichia Enterococcus spp., Pinworms spp., Enterococcus spp., Enterovirus spp., Rickettsia typhi, Parvo Virus B19, human herpesvirus type 6, human herpesvirus type 7, thrombocytopenic fluke, liver Leeches and giant liver flukes, FFI prions, Filarioidea, Clostridium perfringens, Fusobacteria Clostridium, Clostridium perfringens, other Clostridium species, Geotrichum candidum, GSS puri On, Giardia lamblia, Burkholderia mallei, Gnathostoma nematode, Gnathostoma nigricans, Neisseria gonorrhoeae, Borrelia granulomatosa, Streptococcus pyogenes, Streptococcus agarica Cutiaria, Haemophilus influenzae, enteric viruses, most Coxsackie A viruses, enteric Enterovirus type 71, Sin Nombre virus, Helicobacter pylori, Escherichia coli O158: H7, Bunyaviridae, Hepatitis A virus, Hepatitis B virus, Hepatitis C virus, D Hepatitis virus, Hepatitis E virus, Herpes simplex virus type 1, Herpes simplex virus Type 2, Histoplasma capsulatum, duodenal hookworm, American hookworm, influenza Bacteria, Boca Human Virus, Ehrlichia ewingii ), Anaplasma phagocytophilum, human metapneumovirus, Ehrlichia chaffeensis, human papillomavirus, human parainfluenza virus, streak Worms, Tapeworms, Epstein-Barr virus, Orthomyxoviridae, Isospora Isospora belli, Kingella kingae ngae), Klebsiella pneumoniae, Klebsiella ozena, Klebsiella rhinoscherreromortis (Klebsiellarhinoscleromotis), Couloupion, Lackey Leishmaniasis virus, Legionella pneumophila, Legionella pneumophila, Leishmaniasis virus Mycobacterium leprosum and Mycobacterium lepromatosis m lepromatosis), Leptospira, Listeria, Borreliosis and other Borrelia species, Wuchereria bancrofti and Brugia malayi, lymphocytic choriomeningitis virus (L CMV, Plasmodium genus, Marburg virus measles virus, Burkholderia pseudomallei , Neisseria meningitidis, Fluke yokogawai, Microsporidia, Molluscum contagiosum virus (MCV), Mumps virus Rickettsia typhi, Mycoplasma pneumoniae, various bacteria (Actinomycetes (mycetoma) and fungi (mycetoma), parasitic fly larvae of the Diptera order, Chlamydia trachomatis and Neisseria gonorrhoeae, vCJD prion, Nocardia asteroides and other Nocardia species, convoluted Insects, Blastomyces brasiliensis, Paragonimus and other Paragonimus species, Pasteurella species, and Atamaji Body lice, Phthiruspubis, Bordetella pertussis, Streptococcus pneumoniae, Pneumocystis cysticercosis, Poliovirus, Prevotella spp., Naegleria Amoeba, JC virus, Psittacosis, Chlamydia, Coxiella burnetii, Rabies Viruses, bead chain E. coli and rat bite fever spirochetes, respiratory respiratory syncytial virus , Rhinosporidium sayberi, Rhinovirus, Rickettsia spp., Rickettsia mites, Rif Tovalley fever virus, Rocky Mountain spotted fever rickettsia, rotavirus, rubella virus, monkey Monera, atypical pneumonia coronavirus, scabies mites, Schistosoma, Shigella, Varicella zoster virus Virus, Variola major or minor, Sporothrix schenckii, Staphylococcus spp., Staphylococcus aureus Cocci, Streptococcus pyogenes, Strongyloides stercoralis, Syphilis spirochete, Cestodes spp., Clostridium tetani, Ringworm, Trichophyton Tongue rot, ringworm, Epidermophyton floccosum, Trichophyton rubrum and Trichophyton barbae Tinea fungus, Trichophyton rubrum, Holtea werneckii, Ringworm, Malassezia, Roundworm of the Dog and Roundworm of the Cat , Toxoplasma gondii, Trichinella spiralis, Trichomonas vaginalis, Trichuris trichiura, Mycobacterium tuberculosis, Tula hot francis Bacteria, Ureaplasma urealyticum, Venezuelan equine encephalitis virus, Vibrio cholerae, Anaritovirus, West Nile virus, Trichosporonium spp., Yersinia pseudotuberculosis, Yersinia enterocolitica, Yellow fever Disease viruses, Mucorales (mucormycosis) and insect mesh mold (entomophthora infection), Pseudomonas aeruginosa , Campylobacter fetus (Vibrio), Aeromonas bacteria, Edwardsiella spp. Tarda , Yersinia pestis, Shigella dysenteriae, Shigella sonnei, Salmonella typhimurium, Treponema pertussis Nue, Treponema carateneum, Fensenburgdorferi, Borrelia burgdorferi feri, Leptospira hemorrhagic jaundice, Pneumocystis carinii, Brucella abortus, Brucella abortus, Malta fever, Mycoplasma spp., Rickettsia typhi, Rickettsia tsutsugamushi, Kura Mysia spp., pathogenic fungi (Aspergillus fumigatus, Candida albicans, Histolytica Toplasma capsulatum); protozoa (Entamoeba histolytica, Trichomonas vaginalis, Trichomonas vegetans, Trypanosoma gambiens, Trypanosoma rhodesiense, Leishmania donovani, Leishmania tropicalis, Leishmania brasiliensis, Pneumocystis carinii pneumonia, Plasmodium vivax Plasmodium, Plasmodium falciparum, malignant malaria); or helminths (Schistosoma japonicum, Schistosoma mansoni) These include, but are not limited to, blood flukes, Schistosoma haematobium and hookworms.

[0165] Other antibodies as cell-binding ligands for use in the present invention for the treatment of viral diseases Examples of antibodies include, but are not limited to, antibodies against pathogenic virus antigens, and the pathogenic virus Examples of viruses include, but are not limited to, viruses from the family Poxyirida e), Herpesviridae, Adenoviridae, Papovaviridae, Enteroviridae , Picornaviridae, Parvoviridae, Reoviridae, Retroviridae, Influenza influenza virus, parainfluenza virus, mumps, measles, respiratory syncytial Viruses, rubella, arboviruses, rhabdoviruses, arenaviridae, non-A / non-B hepatitis inflammation viruses, rhinoviruses, coronaviruses, rotaviruses, tumor viruses [e.g. HBV (hepatocellular carcinoma), HPV (cervical cancer, anal cancer), Kaposi's sarcoma-associated herpes virus ( Kaposi's sarcoma), EB virus (nasopharyngeal carcinoma, Burkitt's lymphoma, primary central nervous system lymphoma tumor), MCPyV (Merkel cell carcinoma), SV40 (Simian virus 40), HCV (hepatitis C virus) caused by viruses such as HTLV-I (adult T-cell leukemia / lymphoma); Immune diseases: [e.g., human immunodeficiency virus (AIDS)], CNS viruses: [ For example, JCV (progressive multifocal leukoencephalopathy), MeV (subacute sclerosing panencephalitis), LCV (liver vein leukoencephalopathy), Lymphocytic choriomeningitis), arboviral encephalitis, Orthomyxoviridae (presumed) (Encephalitis lethargica), RV (Rabies), Vesicular stomatitis, Herpes virus meningitis, Ramsay Hunt syndrome type II; poliovirus (acute poliomyelitis, post-polio syndrome), HTLV- I (tropical spastic paralysis)]; cytomegalovirus (CMV) retinitis, HSV (herpes simplex virus) Cardiovascular diseases (e.g., CBV (pericarditis, myocarditis)); respiratory / acute nasopharyngeal Headache / viral pneumonia: [EB virus (EBV infection / infectious mononucleosis), cytomegalovirus (HBV) Rovirus, SARS coronavirus (Severe Acute Respiratory Syndrome), Orthomyxovirus Influenza virus A / B / C (influenza / avian influenza) ), Paramyxovirus: Human parainfluenza virus (parainfluenza), R SV (human respiratory syncytial virus), hMPV]; digestive system viruses [MuV (mumps) esophagitis), cytomegalovirus (CMV esophagitis); adenovirus (adenovirus infection) Infection); Rotavirus, Norovirus, Astrovirus, Coronavirus, HBV (type B) Hepatitis virus), CBV, HAV (hepatitis A virus), HCV (hepatitis C virus), HDV (hepatitis D virus), HEV (hepatitis E virus), HGV (hepatitis G virus) )]; urogenital viruses [e.g., BK virus, MuV (mumps)] It can be enjoyed.

[0166] According to a further object, the present invention provides a method for the preparation of a conjugate of the present invention via a crosslinked conjugate and a pharmaceutical and / or a combination of a medicament for treating cancer, an infectious disease, or an autoimmune disease, together with an acceptable carrier, diluent, or excipient. The present invention also relates to pharmaceutical compositions for treating cancer, infectious diseases, and autoimmune diseases. The method may be in vitro, in vivo or ex vivo Examples of in vitro therapy include transfection of target antigens into the target cells (ex vivo). To kill all cells except for the desired mutant that does not express or express the undesired antigen. Examples of ex vivo therapies include treatment of cell cultures to kill mutants that express the virus. Hematopoietic stem cells (HSCs) are treated to kill diseased or malignant cells prior to transplantation (HSCT). This includes returning the cells to the same patient for other purposes, such as treating cancer and autoimmune diseases. for the removal of cancer or lymphoid cells from bone marrow prior to autologous transplantation in Prior to transplantation, T cells and IgG from allogeneic bone marrow or tissue are obtained to prevent graft-versus-host disease. Clinical ex vivo treatment to remove other lymphoid cells can be performed by: After obtaining bone marrow cells from a patient or other individual, the concentration range is approximately 1 pM to 0.1 mM. The conjugate of the present invention is added to a serum-containing medium and incubated at 37°C for 30 minutes to about 48 hours. The exact concentration conditions and incubation time (= dose) can be easily determined by an experienced clinician. After the culture is completed, the bone marrow cells are washed with a serum-containing medium and then injected intravenously or by other known methods. During the bone marrow cell procurement and reinfusion treatment, the patient may not undergo other treatments (e.g., If the patient has received chemotherapy or total body irradiation after treatment, the processed bone marrow cells may be used with standard medical equipment. It is frozen and preserved using liquid nitrogen.

[0167] For in vivo clinical applications, the conjugates of the present invention via the linker may be administered in the form of a solution or injection. It is supplied in the form of a lyophilized solid that can be redissolved in sterile water for appropriate co-administration. An example of a method for administering the conjugate is as follows: The conjugate is administered intravenously over a period of 8 to 20 weeks. Dissolve the bolus dose in 50-500 mL of saline and administer in saline. Human serum albumin can be added to the solution (e.g., 0.5–5 ml of concentrated human serum albumin). The drug dosage is approximately 50 μg to 20 mg / kg body weight per week. and is administered intravenously (injection dose ranges from 10 μg to 200 mg / kg). After completing 0 weeks of treatment, patients can receive a second course of treatment. Detailed treatment methods, including route, vehicle, diluent, dosage, and duration of treatment, should be determined by an experienced surgeon. It can be determined that:

[0168] Treating diseases by selectively killing cell populations using in vivo or ex vivo methods Examples include any type of cancer, autoimmune diseases, transplant rejection, and infectious diseases (viruses). (including viruses, bacteria or parasites).

[0169] Several factors affect the amount of conjugated drug required for the desired biological effect. The characteristics of the compound, the efficacy and bioavailability of the conjugate drug, the type of disease, the patient's race, the patient's The administration schedule and dosage are determined by taking these factors into account. The route is determined.

[0170] Generally, the conjugates of the present invention via the linker are used at a concentration of 0.1 to 10% w / v. It may be a formulation for parenteral administration dissolved in a physiological buffer so as to contain the compound. Typical dosage ranges are 1 μg / kg body weight to 0.1 g / kg body weight per day, preferably The recommended dose range is 0.01 mg / kg to 20 mg / kg of body weight per day, or The preferred drug dosage depends, for example, on the type and stage of progression of the disease or disorder. the overall health of the individual patient, the relative biological activity of the selected drug, and the compound Dosage form, mode of administration (intravenous, intramuscular, or other), and dosage of the drug in the selected mode of administration Pharmacokinetic properties, as well as the rate of administration (single injection or continuous infusion) and schedule of administration (constant The dose depends appropriately on variables such as the frequency of administration over time.

[0171] The conjugates of the present invention via the linker may also be administered in unit dosage amounts, where "unit dosage" refers to "Quantity" means a single dose administered to one patient and is to be used in simple and convenient packaging. The active conjugates can be physically and / or therapeutically effective as active conjugates themselves or as pharmaceutically acceptable compositions as described below. It maintains a stable dosage and chemically stable unit dose. The dosage range is 0.01 to 100 mg / kg body weight. General guidance is: The unit dosage ranges from 1 to 3000 mg per day, week, biweekly, or month. The dosage is preferably 1 mg to 500 mg administered 1 to 4 times a week, and more preferably Preferably, 1 mg to 100 mg is administered once a week. The pharmaceutical composition may be prepared by adding one or more pharmaceutically acceptable excipients to the pharmaceutical composition. The unit dose of the drug may be packaged as a tablet, simple capsule, or soft capsule for oral administration. for intranasal administration as a powder, nasal drops, or aerosol; or for dermal administration For administration, the agent may be administered as an ointment, cream, lotion, gel, or spray or skin patch. It can be administered as follows.

[0172] Drugs / Cytotoxic Agents

[0173] Drugs that can be conjugated to cell-binding molecules in the present invention include small molecules, including cytotoxic agents. A small molecule drug can be linked to a cell-binding molecule directly or after modification. The molecular weight of the drug may be, for example, 100 to 2500, more preferably 120 to 1500. Organic, inorganic or organometallic compounds are widely used. For a better definition of small molecule drugs, Reference is made to WO05058367A2 and U.S. Pat. No. 4,956,303, as well as other documents. The drugs may be any of the known drugs listed above, which are incorporated by reference in their entirety. This includes substances and potential drugs.

[0174] Known drugs include, but are not limited to:

[0175] a) Alkylating agents: Nitrogen mustards: Chlorambucil, chlornaphazine, Cyclophosphamide, dacarbazine, estramustine, ifosfamide, mechlorethamine mechlorethamine oxide hydrochloride, mannomustine, mitobronitol, melphalan , Mitolactol, Pipobroman, Novenbitine, Fenesterine, Prednimustine, Thiotepa, trofosfamide, uracil mustard; CC-1065 (adzelesin, including synthetic analogs of rzeresin and bizeresin; duocarmycins (synthetic analogs, KW-2189 and CBI-TMI; benzodiazepine dimers (pyrrolobenzoxazoles) Diazepine (PBD) or tomaymycin, indolinobenzodiazepines, imidazobe dimers of benzothiazepines or oxazolidinobenzodiazepines . );Nitrosourea compounds: (carmustine, lomustine, chlorozotocin, fotemus Alkyl sulfonates: (busulfan, threosulfan, nimustine, ranimustine) sulfan, improsulfan and piposulfan); triazene (dacarbazine); Gold-containing compounds: (carboplatin, cisplatin, oxaliplatin); aziridines, Benzodopa, carboquone, metholedopa and uredopa; ethyleneimines, and alto Retamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophos methylmelamines including trimethylolmelamine; b) Plant alkaloids: Vinca alkaloids: (vincristine, vinblastine, Vindesine, vinorelbine, navelbine; Taxoids: (paclitaxel, docetaxel cell) and their analogues, maytansinoids (DM1, DM2, DM3, DM4, D M5, DM6, DM7, maytansine, ansamitocin) and their analogues, cryptophycins (selected from cryptophycin 1 and cryptophycin 8); rons, eluterobins, discodermolide, bryostatins, dolostatins , auristatins, tubulysins, cephalostatins; pancratistatin; Rucodictyin;Spongistatin, etc.; c) DNA topoisomerase inhibitors: [Epipodophyllins: (9-aminocamptothecin camptothecin, crisnatol, daunomycin, etoposide, etoposide phosphate Do, irinotecan, mitoxantrone, novantrone, retinoic acid (retinols), Teniposide, topotecan, 9-nitrocamptothecin (RFS 2000); Mitomycin Mitomycins (Mitomycin C) etc. d) Antimetabolites: {[Antifolates: Dihydrofolate reductase inhibitors: (Methotrexate , trimetrexate, denopterin, pteropterin, aminopterin (4-aminopterin) teroic acid), or folic acid analogs); IMP dehydrogenase inhibitors (mycophenolic acid, tiazofurin, ribavirin, EICAR); ribonucleotide reductase inhibitors (hydroxybenzoates) thiourea, deferoxamine)]; [pyrimidine analogs: uracil analogs (ancitabine , azacitidine, 6-azauridine, capecitabine (Xeloda), carmofur, citara vin, dideoxyuridine, doxifluridine, enocitabine, 5-fluorouracil, Floxuridine, raltitrexed (Tomudex); cytosine analogs: (Citara vin, cytosine arabinoside, fludarabine); Purine analogs: (azathioprine, fludarabine Darabine, mercaptopurine, thiamiprine, thioguanine)]; folic acid supplements, fluorine acid}etc.; e) Hormonal therapy agents: {Receptor antagonists: [Antiestrogens: (megestrol, raloxacin, cyclosporine, tamoxifen); LHRH agonists: (goserelin, leuprolide acetate) ); Antiandrogens: (bicalutamide, flutamide, calcitonin, dromedapropionate Stanolone, epithiostanol, goserelin, leuprolide, mepitiostane, nil thamide, testolactone, trilostane, and other androgen inhibitors)]; retinoids Class / deltoid: [Vitamin D3 analogues: (CB1093, EB1089, KH1060, Recalciferol, ergocalciferol; Photodynamic therapy agents: (verteporphyrin phthalocyanines, photosensitizer Pc4, demethoxy-hypocrelin A); cytokines : (Interferon α, interferon γ, tumor necrosis factor (TNF), TNF domain phospho-containing human proteins)]} etc.; f) Kinase inhibitors: BIBW2992 (anti-EGFR / Erb2), imatinib, GEF ritinib, pegaptanib, sorafenib, dasatinib, sunitinib, erlotinib, nilotinib tinib, lapatinib, axitinib, pazopanib, vandetanib, E7080 (anti-VEG FR2), mubritinib, ponatinib (AP24534), bafetinib (INNO-4 06), bosutinib (SKI-606), cabozantinib, vismodegib, iniparib, Ruxolitinib, CYT387, axitinib, tivozanib, sorafenib, bevacizumab , cetuximab, trastuzumab, ranibizumab, panitumumab, ispinesib, etc.; g) Olaparib, Niraparib, Iniparib, Talazoparib, Veliparib, CEP972 2 (Cephalon), E7016 (Eisai), BGB-290 (BeiGene ), or 3-aminobenzamide, PARP inhibitors, etc. h) Antibiotics: Enediyne antibiotics (calicheamicins, calicheamicin γ1, δ1, α1, and β1; dynemicins, including dynemicin A and deoxydynemicin; Speramicin, kedarcidin, C-1027, maduro-peptin, and neocarzinostat Consists of cin chromophores and related enediyne antibiotic chromophores group), aclacinomycins, actinomycin, anthramycin, Xaserin, bleomycins, cactinomycin, carabinieri carabicin, carminomycin, carzinophilin; chromomycins , dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-no Doxorubicin, morpholino-doxorubicin, cyanomorpholino-doxorubicin Rubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin, epirubicin, Esorubicin, idarubicin, marcelomycin, mitomycins, mycophenolic acid , nogalamycin, olivomycins, peplomycin, potfilomycin, puromycin mycin, querramycin, rhodolubicin, streptonigrin, streptozocin, Versigin, ubenimex, zinostatin, zorubicin, etc.; i) Polyketides (acetogenins), bullatacin and bullatacinone; gemcitabine, Poxomicins (including carfilzomib), bortezomib, thalidomide, lenalid amide, pomalidomide, tosedostat, zybrestat, PLX4032, STA- 9090, Stimuvax, Allovectin-7, Zygeba, Probe fluphenazine, ervoy, isoprenylation inhibitors (selected from lovastatin), dopamine action Neurotoxicity (selected from 1-methyl-4-phenylpyridine ions), cell cycle inhibitors Actinomycins (actinomycin D, da cytochrome P450 (selected from the group consisting of bleomycin A2, bleomycin B2, peplomycin), anthracyclines (daunorubicin, doxorubicin (adriamycin), idarubicin, epirubicin methicillin, pirarubicin, zorubicin), mitoxantrone, MDR inhibitors or verapamil, Ca 2+ ATP inhibitors or thapsigargin, histone deacetylases Metabolic enzyme inhibitors (vorinostat, romidepsin, panobinostat, valproic acid, mocetin) Nostat (MGCD0103), Belinostat, PCI-24781, Entinostat SB939, resminostat, gibinostat, AR-42, CUDC-101 , sulforaphane, trichostatin A); thapsigargin, Celecoxib, glitazones, epigallocatechin gallate, disulfiram, salinospo Anti-adrenal drugs (selected from the group consisting of aminoglutethimide, mitotane, and trilostane) Aceglatone; Aldophosphamide glucoside; Aminolevulinic acid; Amsac Phosphorus; Arabinoside, Bestravsil; Bisantrene; Edatrexate; Defofami acetone, demecolcine, diaziquone, elfornithine (DFMO), elfomicin; acetic acid Elliptinium, etocluzid; gallium nitrate; gacytosine; hydroxyurea; Iband Ionate, Lentinan; Lonidamine; Mitoguazone; Mitoxatrone; Mopidamol; Traeline; Pentostatin; Fenamet; Pirarubicin; Podophyllic acid; 2-Ethyl Procarbazine; PSK (registered trademark); Razoxane; Rhizoxin; Schizophrenia Spirogermanium; Tenuazonic acid; Triazicon; 2,2',2''-triazine Chlorotriethylamine; Trichothecenes (T2 toxin, verrucarin A, roridin A, and anguidine); urethane, siRNA, antisense Drugs, nucleolytic enzymes, etc.

[0176] (2) Anti-autoimmune disease drugs: Cyclosporine, cyclosporine A, aminocaproic acid, azathioprine, bromocriptine acetaminophen, chlorambucil, chloroquine, cyclophosphamide, corticosteroids (ambucil, Cinonide, betamethasone, budesonide, hydrocortisone, flunisolide, fluticasone Propionate ester, fluocortodanazol, dexamethasone, triamcinolone cetonide, beclomethasone dipropionate, etc.), DHAE, etanercept, hydroxylase Chloroquine, infliximab, meloxicam, methotrexate, mycophenolate mofetil These include, but are not limited to, fetyl, prednisone, sirolimus, and tacrolimus. stomach.

[0177] (3) Anti-infective drugs: a) Aminoglycosides: Amikacin, astromycin, gentamicins (Netimata) Isin, Sisomycin, Isepamicin), Hygromycin B, Kanamycins (Ami Kashin, Arbekacin, Bekanamycin, Dibekacin, Tobramycin), Neomycin (framycetin, paromomycin, ribostamycin), netilmicin, specti nomycin, streptomycin, tobramycin, verdamycin; b) Amphenicols: azidamphenicol, chloramphenicol, florphen Nicole, thiamphenicol; c) Ansamycins: geldanamycin, herbimycin; d) Carbapenems: biapenem, doripenem, ertapenem, imipenem / cilasta tin, meropenem, panipenem; e) Cephems: Carbacephem (loracarbef), cephacetrile, cefaclor, Cephradine, cefadroxil, cephalonium, cephaloridine, cephalothin or cephalosporin Phalosporin, cephalexin, cephaloglycin, cefamandole, cephapirin , cefatrizine, cefazaflur, cefazedone, cefazolin, cefbuperazone, cef Capen, cefdaloxime, cefepime, cefminox, cefoxitin, cefprozil , cephalosporins, ceftezol, cefuroxime, cefixime, cefdinir, cefdito Ren, cefepime, cefetamet, cefmenoxime, cefodizime, cefonicid, cef Operazon, ceforanide, cefotaxime, cefotiam, cefozopran, cephalexin Cefpiramide, cefpirome, cefpodoxime, cefprozil, Fukinom, cefsulodin, ceftazidime, cefteram, ceftibuten, ceftiolene, ftizoxime, ceftobiprole, ceftriaxone, cefuroxime, cefuzonam, cef Cefoxitin, cefotetan, cefmetazole, oxacephem (flo Moxef, latamoxef); f) Glycopeptides: bleomycin, vancomycin (oritavancin, telavancin) ), teicoplanin (dalbavancin), ramoplanin; g) Glycylcyclines: Tigecycline; h) β-lactamase inhibitors: penams (sulbactam, tazobactam), clavams (clavams) labranic acid); i) Lincosamides: clindamycin, lincomycin; j) Lipopeptides: Daptomycin, A54145, calcium-dependent antibiotics (C D.A.); k) Macrolides: azithromycin, cethromycin, clarithromycin, zirconium Thromycin, erythromycin, flurithromycin, josamycin, ketolide ( Telithromycin, cethromycin), midecamycin, miocamycin, oleandoma Rifamycins (rifampicin, rifampin, rifabutin, rifapentin) rokitamycin, roxithromycin, spectinomycin, spiramycin , tacrolimus (FK506), troleandomycin, telithromycin; l) Monobactams: aztreonam, tigemonam; m) Oxazolidinones: Linezolid; n) Penicillins: Amoxicillin, Ampicillins (Pivampicillin, Hetacillin, Bacampicillin, methampicillin, talampicillin), azidocillin, azlocillin, benzampicillin benzylpenicillin, benzathine benzylpenicillin, benzathine phenoxymethylpenicillin Phosphorin, clometocillin, procaine benzylpenicillin, carbenicillin (calindacillin cloxacillin, dicloxacillin, epicillin, flucloxacillin, mesilina pivmecillinam, mezlocillin, methicillin, nafcillin, oxacillin, penam Penicillin, phenethicillin, phenoxymethylpenicillin, piperacillin, propranolol lopicillin, sulbenicillin, temocillin, ticarcillin; o) Polypeptides: bacitracin, colistin, polymyxin B; p) Quinolones: alatrofloxacin, balofloxacin, ciprofloxacin, Linafloxacin, danofloxacin, difloxacin, enoxacin, enrofloxacin Syn, Floxacin, Garenoxacin, Gatifloxacin, Gemifloxacin, Grepaflo flucloxacin, carbafloxacin, levofloxacin, lomefloxacin, marbofloxacin fluoxacin, moxifloxacin, nadifloxacin, norfloxacin, orbifloxacin ofloxacin, pefloxacin, trovafloxacin, grepafloxacin, Tafloxacin, sparfloxacin, temafloxacin, tosufloxacin, torovaf Loxacin; q) Streptogramins: Pristinamycin, quinupristin / dalfopristin ; r) Sulfonamides: Mafenide, Prontosil, Sulfacetamide, Sulfamethoxazole azole, sulfanilamide, sulfasalazine, sulfisoxazole, trimethoprim , trimethoprim-sulfamethoxazole (cotrimoxazole); s) Steroidal antibacterial agents: fusidic acid, etc.; t) Tetracyclines: doxycycline, chlortetracycline, chromocycline Phosphorus, demeclocycline, lymecycline, meclocycline, methacycline, minocycline Cycline, oxytetracycline, penimepicycline, rolitetracycline, tetracycline tracycline, glycylcyclines (tigecycline, etc.); u) Other types of antibiotics: annonacin, arsphenamine, bactoprenol inhibitors agent (bacitracin), DADAL / AR inhibitor (cycloserine), dictyostatin, Discodermolide, eleutherobin, epothilone, ethambutol, etoposide, Alopenem, fusidic acid, furazolidone, isoniazid, laulimalide, metronidazole ru, mupirocin, mycolactone, NAM synthesis inhibitor (fosfomycin), nitrofuran Toxin, paclitaxel, platensimycin, pyrazinamide, quinupristin / dalphos Pristine, rifampin (rifampin), tazobactamcinidazole, uvaricin; These include, but are not limited to:

[0178] (4) Antiviral drugs: a) Entry / fusion inhibitors: aplaviroc, maraviroc, vicriviroc, gp41(E) fuvirtide), PRO140, CD4 (ibalizumab); b) Integrase inhibitors: Raltegravir, Elvitegravir, Globoidnan A; c) Maturation inhibitors: Bevirimat, Vivicon; d)Neuraminidase inhibitors: oseltamivir, zanamivir, peramivir; e) Nucleosides and nucleotides: abacavir, acyclovir, adefovir, amphotericin Doxovir, apricitabine, brivudine, cidofovir, clevudine, dexerbucitabine bin, didanosine (DDI), elvucitabine, emtricitabine (FTC), entecavir vir, famciclovir, fluorouracil (5-FU), 3'-fluoro substituted 2',3 '-deoxynucleoside analogue (3'-fluoro-2',3'-dideoxythymidine ( FLT) and 3'-fluoro-2',3'-dideoxyguanosine (FLG) ), fomivirsen, ganciclovir, idoxuridine, lamivudine ( 3TC), L-nucleosides (β-L-thymidine and β-L-2'-deoxycytidine) ), penciclovir, rasvir, ribavirin, stampidine, Stavudine (d4T), taribavirin (viramidine), telbivudine, tenofovir , trifluridine, valacyclovir, valganciclovir, zalcitabine (ddC), di Dovudine (AZT); f) Non-nucleosides: amantadine, ateviridin, capravirine, diarylpyrimidin (etravirine, rilpivirine), delavirdine, docosanol, emivirine, efavirdine Virenz, foscarnet (phosphoryl formate), imiquimod, interferon alpha, Robinson Lidl, rhodenosine, methisazone, nevirapine, NOV-205, peginterferon alpha , podophyllotoxin, rifampicin, rimantadine, resiquimod (R-848), Romantadine; g) Protease inhibitors: amprenavir, atazanavir, boceprevir, darunavir , fosamprenavir, indinavir, lopinavir, nelfinavir, pleconaril, Tonavir, saquinavir, telaprevir (VX-950), tipranavir; h) Other antiviral drugs: Abzyme, Arbidol, Calanolide A, Seragenin, Cyanovirin-N, Diarylpyrimidine, Epigallocatechin Gallate (EGCG), Phosphorus Carnet, Griffithsin, taribavirin (viramidine), hydroxyurea, KP- 1461, Miltefosine, Pleconaril, Portmanteau inhibitors, Ribavirin, Sericin Crib;

[0179] 5) Drugs used for conjugation via the bridge linkers of the present invention include radioisotopes. Examples of radioisotopes (radionuclides) include: 3 H, 11 C. 14 C. 18 F, 32 P, 35 S, 64 Cu, 68 Ga, 86 Y, 99 Tc, 111 In, 123 I, 124 I , 125I, 131 I, 133 Xe, 177 Lu, 211 At, and 213 Bi mentioned Radiolabeled antibodies are very useful in receptor-targeted imaging experiments, or For example, the invention of antibody-drug conjugates (Wu et al. (2005) Nature Biotechnology 23(9):1137 -1146), which can be used for direct relative targeting therapy, cell-binding molecules, e.g. The antibody may be conjugated, chelated, or otherwise complexed with a cross-linker of the present invention, as described above. This labeling technique is described in the Current Protocol. s in Immunology, Volumes 1 and 2, Coligen et al, Ed. Wiley-Interscience, New York k, NY, Pubs. (1991). Chelating agents that can form complex metal complexes include , DOTA, DOTP, DOTMA, DTPA, and TETA (Macrocyclics, Dallas, Tex.) are included.

[0180] 6) Pharmaceutically acceptable salts, acids, or derivatives of any of the above drugs.

[0181] In another embodiment, the drug of formula (II) and / or (IV) binds to a cell-binding molecule and targets the Chromophores that can be conjugated to detect, monitor, or study interactions with cells Chromophore molecules can be UV light, fluorescent light, IR light, near-IR light, or visible light. chromophores are compounds that have the ability to absorb certain types of light, such as xanthophores, red The classes or subclasses of chromatophores, iridophores, leucophores, melanophores, cereuphores, and fluorophores The phosphor comprises a fluorescent chemical that re-emits light in light; a visible light transmitting molecule; Class or subclass of luminescent molecules; Class or subclass of luminescent molecules or subclass; a class or subclass of luciferin compounds.

[0182] Chromophore molecules include, but are not limited to, xanthene derivatives (fluorescein, rhodopsin, amine, Oregon Green, Eosin, and Texas Red); cyanine derivatives (cyanine , indocarbocyanines, oxacarbocyanines, thiacarbocyanines, and merocyanines squalene derivatives and ring-substituted squaraines (Seta, SeTau, and Squa) including re dyes); naphthalene derivatives (dansyl and prodan derivatives); coumarin derivatives oxadiazole derivatives (pyridyloxazole, nitrobenzoxadiazole, and benzoxadiazoles; anthracene derivatives (DRAQ5, DRAQ7, and CyT Anthraquinones, including RAK Orange; pyrene derivatives (e.g., Cascade Blue) ;Oxazine derivatives (Nile Red, Nile Blue, Cresyl Violet, Oxazine 170, etc.). Acridine derivatives (proflavine, acridine orange, acridine yellow) Arylmethine derivatives (auramine, crystal violet, malachite glycol) Tetrapyrrole derivatives (porphine, phthalocyanine, bilirubin) and other non-tannin compounds The phosphor can be selected from a group consisting of phosphorus, phosphate, phosphate-containing organic phosphors, and phosphate-containing organic phosphors.

[0183] Alternatively, the chromophore molecule is selected from any analogues and derivatives of the following fluorophore compounds: CF dye (Biotium), DRAQ and CyTRAK probes (Bio-Status ), BODIPY (Invitrogen), Alexa Fluor (Invitrogen), DyLig ht Fluor (Thermo Scientific, Pierce), Atto and Trance (Sigma A ldrich), FluoProbes (Interchim), Abberior dye (Abberior), DY and MegaStokes dyes (Dyomics), SulfoCy dyes (Cyandye), Hi Lyte Fluor (AnaSpec), Seta, SeTau and Square dyes (SETA BioMedicals), Quasar and Cal Flour dyes (Biosearch Technologies) , SureLight dyes (APC, RPEPerCP, phycobilisomes) (Columbia Biosciences), APC, APCXL, RPE, BPE (Phyco-Biotech).

[0184] Examples of widely used fluorophore compounds that can react or conjugate with the conjugates of the present invention include: , allophycocyanin (APC), aminocoumarin, APC-Cy7 conjugate, BODIP Y-FL, Cascade Blue, Cy2, Cy3, Cy3.5, Cy3B, Cy5, Cy5 .5, Cy7, Fluorescein, FluorX, Hydroxycoumarin, Lissamine rhodamine B, Lucifer Yellow, Methoxycoumarin, NBD, Pacific Blue, P Acific Orange, PE-Cy5 conjugate, PE-Cy7 conjugate, PerCP, R-Phycoerythrin (PE), Red 613, Seta-555-azide, Seta- 555-DBCO, Seta-555-NHS, Seta-580-NHS, Seta- 680-NHS, Seta-780-NHS, Seta-APC-780, Seta-P erCP-680, Seta-R-PE-670, SeTau380-NHS, SeTa u405-maleimide, SeTau405-NHS, SeTau425-NHS, SeT Examples include au647-NHS, Texas Red, TRITC, TruRed, and X-rhodamine. It can be obtained.

[0185] Fluorophores that can be linked to the conjugates of the invention for the study of nucleic acids or proteins The compound is selected from the following compounds or derivatives thereof: 7-AAD (7-aminoadenine dihydrochloride) Cutinomycin D, CG selective), acridine orange, chromomycin A3, CyTR AK Orange (Biostatus, red excitation dark), DAPI, DRAQ5, DRAQ7, Ethidi ammonium bromide, Hoechst 33258, Hoechst 33342, LDS751, Mithramycin Synthon, propidium iodide (PI), SYTOX Blue, SYTOX Green, SYTO X Orange, Thiazole Orange, TO-PRO: Cyanine Monomer, TOTO-1, TO -PRO-1, TOTO-3, TO-PRO-3, YOseta-1, YOYO-1. Fluorescent dyes that can be linked to the conjugates of the invention for cell studies include the following compounds: Selected from its derivatives: DCFH (2'7'dichlorodihydro-fluorescein, acid Dihydrorhodamine 123 (oxidized form), DHR (dihydrorhodamine 123, oxidized form, light catalyzes oxidation), Fluo -3 (AM ester, pH > 6), Fluo-4 (AM ester, pH 7.2), Ind o-1 (AM ester, low / high calcium (Ca 2+ )), and SNARF (pH 6 / 9 ) Preferred conjugates that can be linked to the conjugates of the present invention for protein / antibody studies are: The fluorescent dye is selected from the following compounds or their derivatives: allophycocyanin (APC) ), AmCyan1 (tetramer, Clontech), AsRed2 (tetramer, Clontech), Aza mi Green (monomer, MBL), azurite, B-phycoerythrin (BPE), celeriac Ann, CyPet, DsRed monomer (Clontech), DsRed2 ("RFP", Clontech) ch), EBFP, EBFP2, ECFP, EGFP (weak dimer, Clontech), Emerald (weak dimer, Invitrogen), EYFP (weak dimer, Clontech), GFP (S65A mutant ), GFP (S65C mutant), GFP (S65L mutant), GFP (S65T mutant) , GFP (Y66F mutant), GFP (Y66H mutant), GFP (Y66W mutant), GFP uv , HcRed1, J-Red, Katyusha, Kusabira Orange e (monomer, MBL), mCFP, mCherry, mCitrine, Midrish i Cyan (dimer, MBL), mKate (TagFP635, monomer, Evrogen), m Keima-Red (monomer, MBL), mKO, mOrange, mPlum, mRas pberry, mRFP1 (monomer, Tsien Lab), mStrawberry, mTFP1, mTurquoise2, P3 (phycobilisome complex), Peridi nin Chlorophyll (PerCP), R-Phycoerythrin (RPE), T -Sapphire, TagCFP (dimer, Evrogen), TagGFP (dimer, Evrog en), TagRFP (dimer, Evrogen), TagYFP (dimer, Evrogen), tdTo mato (tandem dimer), Topaz, TurboFP602 (dimer, Evrogen), TurboFP635 (dimer, Evrogen), TurboGFP (dimer, Evrogen), T urboRFP (dimer, Evrogen), TurboYFP635 (dimer, Evrogen), Nuss, native GFP, YPet, Zs Green 1 (tetramer, Clontech), Zs Yellow 1 (tetramer, Clontech).

[0186] An example of the structure of an antibody-chromophore molecule conjugate via a bridge linker is Ac01 shown below: Ac02, Ac03, Ac04, Ac05, Ac06, Ac07, Ac08, and Ac0 9 include: [ka] TIFF0007643734000069.tif160170

[0187] During the ceremony, [ka] represents a single or double bond; mAb is an antibody, preferably a monoclonal antibody; n , m1, m2, X1, X2, X3, R1, R2, R3, L1, L2, and L are each a group represented by formula (I) and (II).

[0188] In another embodiment, the drugs of Formula (II) and Formula (III) when administered to a mammal It may also be a polyalkylene glycol, which is used to extend the half-life of the cell-binding molecule. The real alkylene glycols include poly(ethylene glycol) (PEG), poly(propylene glycol) ethylene glycol), and copolymers of ethylene oxide and propylene oxide. Particularly preferred is PEG, and even more particularly preferred is PEG. The most common is monofunctional activated hydroxyPEG (e.g., hydroxyPEG-monocarboxylic acid). Reactive esters, hydroxyPEG-monoaldehydes, hydroxyPEG-monoamines, HydroxyPEG-monohydrazide, HydroxyPEG-monocarbazate, Hydroxy PEG-monoiodoacetamide, hydroxyPEG-monomaleimide, hydroxyPE G-monoorthopyridyl disulfide, hydroxyPEG-monoorthopyridyl disulfide HydroxyPEG-monoxime, HydroxyPEG-monophenylcarbonate , Hydroxyl PEG-monophenylglyoxal, Hydroxyl PEG-monothiazo Lysine-2-thione, Hydroxyl PEG-monothioester, Hydroxyl PEG-mono Thiol, hydroxyl PEG-monotriazine, and hydroxyl PEG-monovinyl The hydroxy PEGs are activated at a single terminus, including hydroxyl sulfones.

[0189] In such embodiments, the polyalkylene glycol has a viscosity of from about 10 Daltons to about 20 0 kDa, preferably about 88 Da to about 40 kDa; and more preferably, the two branches each have a molecular weight of about 88 Da to about 40 kDa; and In one particular embodiment, the polyalkylene glycol is Poly(ethylene) glycol, about 10 kDa; about 20 kDa, or about 40 kDa In certain embodiments, the PEG has a molecular weight of 10 kDa (linear or branched), PEG 20 kDa (linear or branched), or PEG 40 kDa (linear or branched). Numerous U.S. patents, such as U.S. Patents 5,428,128; 5,621,039; 5,622,986; 5,643,575; 5,728,56 0; 5,730,990; 5,738,846; 5,811,076; 5,824,701; 5,840,900; 5,880,131; 5,900,402; 5,902,588; 5,919,455; 5,951,974; 5,965,119; 5,965,566; 5,969,040; 5,981,709; 6,0 11,042; 6,042,822; 6,113,906; 6,127,355; 6,132,713; 6,177,087, and 6,180,095 Preparation of linear or branched "non-antigenic" PEG polymers and their derivatives or conjugates The structure of the antibody-polyalkylene glycol conjugate via a bridge linker is The following Pg01, Pg02, Pg03, Pg04, Pg05, Pg06, and Pg07 Some examples include:

[0190] [ka] TIFF0007643734000072.tif84157

[0191] In the formula, mAb is an antibody; R' is H or CH3; m3 is an integer of 1 to 5000. R3 is OH, H, or R1; [ka] represents a single bond or a double bond; m1, m2, n, L1, L2, X1, X2, R1, R2, and R3 have the same definition as in formulas (I) and (II); Alternatively, it may be H, OH, OCH3, or OC2H5; p is 1 to 2000; Dru g1 has the same definition as in formula (III).

[0192] In yet another embodiment, a compound suitable for conjugation to a cell-binding molecule via a bridge linker of the present application is Preferred cytotoxic agents are tubulysins, maytansinoids, taxanoids (taxanoids), compounds), CC-1065 analogs, daunorubicin and doxorubicin compounds, benzodiazepines Zepine dimers (e.g., pyrrolobenzodiazepines (PBDs), tomaymycin, anthraquinones, mycin, indolinobenzodiazepines, imidazobenzothiadiazepines, or oxazolidinone Dimers of zolidinobenzodiazepines, calicheamicins and enediyne antibiotics , actinomycin, azaserine, bleomycin, epirubicin, tamoxifen , idarubicin, dolastatins, auristatins (e.g., monomethyl auristatin Auristatin E, MMAE, MMAF, Auristatin PYE, Auristatin TP, Auristatin 2-AQ, 6-AQ, EB (AEB), and EFP (AEFP), duocalmycin steroids, geldanamycins, methotrexates, thiourea Tepa, vindesines, vincristines, hemiasterins, nazumamides (nazum amides), microginins, radiosmins iosumins, alterobactins, microspores Microsclerodermins, theonellamides onellamides), esperamicins, PNU- 159682, and their analogs and derivatives.

[0193] Tubulysins, which are preferred compounds for the conjugates of the present invention, are well known in the art. , can be extracted from natural products according to known methods or synthesized by known methods ( For example: Balasubramanian, R.; et al. J. Med. Chem., 2009, 52, 238-240. Wipf, P.; et al. Org. Lett., 2004, 6, 4057-4060. Pando, O.; et al. J. Am. Chem. Soc., 201 1, 133, 7692-7695. Reddy, J. A.; et al. Mol. Pharmaceutics, 2009, 6, 1518-1525. Raghavan, B.; et al. J. Med. Chem., 2008, 51, 1530-1533. Patterson, A. W.; et al . J. Org. Chem., 2008, 73, 4362-4369. Pando, O.; et al. Org. Lett., 2009, 11 (2 4), pp 5567-5569. Wipf, P.; et al. Org. Lett., 2007, 9 (8), 1605-1607. Fries tad, G. K.; Org. Lett., 2004, 6, pp 3249-3252. Hillary M. Peltier, H. M.; et al. J. Am. Chem. Soc., 2006, 128, 16018-16019. Chandrasekhar, S.; et al. J. Org. Ch em., 2009, 74, 9531-9534. Liu, Y.; et al. Mol. Pharmaceutics, 2012, 9, 168-175. Friestad, G. K.; et al. Org. Lett., 2009, 11, 1095-1098. Kubicek, K.; et al., An gew Chem Int Ed Engl, 2010. 49: p. 4809-12. Chai, Y.; et al., Chem Biol, 2010, 1 7: 296-309. Ullrich, A.; et al., Angew Chem Int Ed Engl, 2009, 48, 4422-5. Sani, M.; et al. Angew Chem Int Ed Engl, 2007, 46, 3526-9. Domling, A.; et al., Ang ew Chem Int Ed Engl, 2006. 45, 7235-9. Patent applications: Zanda, M. ; et al, C an. Pat. Appl. CA 2710693 (2011). Chai, Y.; et al. Eur. Pat. Appl. 2174947 (2 010), WO 2010034724. Leamon, C.; et al, WO 2010033733, WO 2009002993. Ellman, J .; et al, WO 2009134279; WO 2009012958, US appl. 20110263650, 20110021568, Mats chiner, G.; et al, WO 2009095447.Vlahov, I.; et al, WO 2009055562, WO 2008112873 . Low, P.; et al, WO 2009026177. Richter, W., WO 2008138561. Kjems, J.; et al, W O 2008125116. Davis, M.; et al, WO 2008076333. Diener, J.; et al, U.S. Pat. Appl . 20070041901, WO 2006096754. Matschiner, G.; et al, WO 2006056464. Vaghefi, F.; et al, WO 2006033913. Doemling, A., Ger. Offen. DE 102004030227; WO 200400532 7; WO 2004005326; WO2004005269. Stanton, M.; et al, US Pat. Appl. Publ. 200402 49130. Hoefle, G.; et al, Ger. Offen. DE 10254439; DE 10241152; DE 10008089. Le ung, D.; et al, WO 2002077036. Reichenbach, H.; et al, Ger. Offen. DE 19638870; Wolfgang, R.; US 20120129779, Chen, H., US appl. 20110027274.). with cell-binding molecules Preferred structures of tubulysins for use in the treatment of cancer are described in PCT / IB2012 / 053554 is described in.

[0194] An example of the structure of the antibody-tubulysin analog conjugate via the linker of this patent is shown below. Below are T01, T02, T03, T04, T05, T06, T07, T08, T09, and T1 0, and T11:

[0195] [ka] TIFF0007643734000075.tif228158TIFF0007643734000076.tif222166TIFF0007643734000077.tif108161

[0196] where mAb is an antibody or cell-binding molecule; n, m1, m2, Drug1, X1, X 2, L1, L2, L3, R1, R2, R3, R4, and R5 are represented by the formulas (I) and (II) R1, R2, R3, and R4 are preferably each independently H, C 1-C8 linear or branched alkyl, aryl, heteroaryl, heteroalkyl, alkylcycloalkyl, ester, ether, amide, amine, heterocycloalkyl, or acyloxyamine; or a peptide containing 1 to 8 amino acids, or a peptide having the structural formula (O CH2CH2) p or (OCH2CH(CH3)) p (p: an integer between 0 and approximately 2000) and the polyethyleneoxy unit is a unit of the following two R: R1R2, R2R3, R1R3, or R3 and R4 are alkyl, aryl, heteroaryl, heteroalkyl, or alkylcycloalkyl. X3 can form a 3- to 8-membered ring of alkyl groups; X3 can be H, CH3, or X 1’ R1 ’ wherein X 1’ is NH, N(CH), NHNH, O, or S, and R 1’ teeth , H or C1-C8 linear or branched alkyl, aryl, heteroaryl, heteroaromatic Z3 is H, OP(O) (OM1)(OM2), OCH2OP(O)(OM1)(OM2), OSO3M1, R1 , or O-glycosides (glucosides, galactosides, mannosides, glucuronosides / glucuronosides) Nidos, allosides, fructosides, etc.), NH-glycosides, S-glycosides or CH2-glycosides is; [ka] represents a single bond or a double bond; M1 and M2 each independently represent H, Na, K, Ca, Mg, NH4, NR1R2R3; 1’ are described throughout this patent It may also be a cytotoxic agent.

[0197] The calicheamicins and related enzymes preferred in the cell-binding molecule-drug conjugates of the present application The lendiine antibiotics are described in Nicolaou, KC et al, Science 1992, 256, 1172-1178; Proc. Natl. Acad. Sci USA. 1993, 90, 5881-5888), US Pa tent Nos. 4,970,198; 5,053,394; 5,108,912; 5,264,586; 5,384,412; 5,606,040; 5,71 2,374; 5,714,586; 5,739,116; 5,770,701; 5,770,710; 5,773,001; 5,877,296; 6,015,5 62; 6,124,310; 8,153,768. Antibody-calicheamicin analog covalent bonds via this bridge linker. Examples of the structure of the character include C01 and C02 below.

[0198] [ka]

[0199] where mAb is an antibody or cell binding molecule; [ka] n, m1, X1, L1, L2, and R1 have the same definitions as in formulas (I) and (II); R 1’ and R 3’ are each independently H or C1-C6 linear or branched alkyl. p is 0 to 2000; 1’ The cytotoxicity described throughout this patent It can also be a drug.

[0200] Maytansin compounds, including maytansinol and its analogues, are preferably used in the present invention. Nos. 4,256,746, 4,361,650, 4 ,307,016, 4,294,757, 4,294,757, 4,371,533, 4,424,219, 4,331,598, 4,450,254, 4,36 4,866, 4,313,946, 4,315,929 4,362,663, 4,322,348, 4,371,533, 4,424,219, 5,208,02 0, 5,416,064, 5,208,020; 5,416,064; 6,333.410; 6,441,163; 6,716,821, 7,276,497, 7,301,019, 7,303,749, 7,368,565, 7,411,063, 7,851,432, and 8,163,888. An example of the structure of a bond-mediated antibody-maytanosinoid conjugate is My01, My02, and My03.

[0201] [ka] TIFF0007643734000082.tif93161

[0202] where mAb is an antibody or cell binding molecule; [ka] n, m1, X1, L1, L2, and R1 have the same definitions as in formulas (I) and (II); R 1’ and R 3’ are each independently H or C1-C6 linear or branched alkyl. p is 0 to 2000; and R 1’ The cytotoxicity described throughout this patent It can also be a drug.

[0203] Paclitaxel (Taxol) is preferred for conjugation via the bridge linkers of this patent. Taxanes, cytotoxic natural products including taxanes and their semisynthetic derivative, docetaxel (taxane) The tails and their analogs are described in K. C. Nicolaou et al. l., J. Am. Chem. Soc. 117, 2409-2420, (1995); Ojima et al, J. Med. Chem. 39:3889 -3896 (1996); 40:267-278 (1997); 45, 5620-5623 (2002); Ojima et al., Proc. Natl. Acad. Sci., 96:4256-4261 (1999; Kim et al., Bull. Korean Chem. Soc., 20, 1389-1 390 (1999); Miller, et al. J. Med. Chem., 47, 4802-4805(2004); U.S. Patent No. 5,47 5,011 5,728,849, 5,811,452; 6,340,701; 6,372,738; 6,391,913, 6.436,931; 6,589,97 9; 6,596,757; 6,706,708; 7,008,942; 7,186,851; 7,217,819; 7,276,499; 7,598,290; and 7,667,054.

[0204] As an example of the structure of the antibody-taxane conjugate via the linker of the present invention, the following Tx0 1, Tx02, and Tx03.

[0205] [ka] TIFF0007643734000085.tif95151

[0206] where mAb is an antibody or cell binding molecule; [ka] represents a single or double bond; n, m1, X1, L1, L2, and R1 are groups represented by the formula (I) and (II) has the same definition as R 1’ and R 3’ are each independently H or C1-C6 is a linear or branched alkyl; p is 0 to 2000; and R 1’ The patent It may also be Drug 1, a cytotoxic agent described throughout.

[0207] CC-1065 analogs and Duocarmycin analogs are also Also preferred for use in conjugation with the bridge linkers of the present invention. CC-1065 analogs and Examples of duocarmycin analogues, as well as their synthesis, are described in the following documents: For example, Warpehoski, et al, J. Med. Chem. 31:590-603 (1988), D. Boger et al., J. Or Chem; 66; 6654-6661, 2001; U.S. Patents: 4169888, 4391904, 4671958, 4816567, 4912227, 4923990, 4952394, 4975278, 4978757, 4994578, 5037993, 5070092, 5084468, 5101038, 5117006, 5137877, 5138059, 5147786, 5187186, 5223409, 5225539, 528851 4, 5324483, 5332740, 5332837, 5334528, 5403484, 5427908, 5475092, 5495009, 55301 01, 5545806, 5547667, 5569825, 5571698, 5573922, 5580717, 5585089, 5585499, 5587 161, 5595499, 5606017, 5622929, 5625126, 5629430, 5633425, 5641780, 5660829, 566 1016, 5686237, 5693762, 5703080, 5712374, 5714586, 5739116, 5739350, 5770429, 57 73001, 5773435, 5786377 5786486, 5789650, 5814318, 5846545, 5874299, 5877296, 58 77397, 5885793, 5939598, 5962216, 5969108, 5985908, 6060608, 6066742, 6075181, 6 103236, 6114598, 6130237, 6132722, 6143901, 6150584, 6162963, 6172197, 6180370, 6194612, 6214345, 6262271, 6281354, 6310209, 6329497, 6342480, 6486326, 651210 1, 6521404, 6534660, 6544731, 6548530, 6555313, 6555693, 6566336, 6,586,618, 65 93081, 6630579, 6,756,397, 6759509, 6762179, 6884869, 6897034, 6946455, 7,049,31 6, 7087600, 7091186, 7115573, 7129261, 7214663, 7223837, 7304032, 7329507, 7,329 ,760, 7,388,026, 7,655,660, 7,655,661, 7,906,545, and 8,012,978. An example of the structure of an antibody-CC-1065 analog conjugate via a ligand is CC01, Examples include CC02 and CC03.

[0208] [ka]

[0209] where mAb is an antibody; Z3 is H, PO(O)(OM1)(OM2), SO3M1 , CH2PO(OM1)(OM2), CH3N(CH2CH2)2NC(O)-, O(C H2CH2)2NC(O)-, R1, or glycoside; X3 is O, NH, NHC(O) , -OC(O), -C(O)O, or R1, or absent; [ka] represents a single or double bond; n, m1, m2, [ka] X1, X2, R1, and R2 are the same as defined in formulas (I) and (II); R 1’ and R 3’ are each independently H or C1-C6 linear or branched alkyl. p is 0 to 2000; and R 1’ The cytotoxicity described throughout this patent Drug 1 may also be used.

[0210] Daunorubicin / doxorubicin analogs are also suitable for conjugation via the bridge linkers of this patent. Its preferred structure and synthesis method are exemplified below: Hurwitz, E., et al. ., Cancer Res. 35, 1175-1181 (1975). Yang, HM, and Reisfeld, RA, Proc. Nat. l. Acad. Sci. 85, 1189-1193 (1988); Pietersz, CA, E., et al., E., et al.," C ancer Res. 48, 926-9311 (1988); Trouet, et al., 79, 626-629 (1982); Z. Brich et al. al., J. Controlled Release, 19, 245-258 (1992); Chen et al., Syn. Comm., 33, 23 77-2390, 2003; King et al., Bioconj. Chem., 10, 279-288, 1999; King et al., J. M ed. Chem., 45, 4336-4343, 2002; Kratz et al., J Med Chem. 45, 5523-33. 2002; tz et al., Biol Pharm Bull. Jan. 21, 56-61 , 1998; Lau et al., Bioorg. Med. Chem 3, 1305-1312, 1995; Scott et al., Bioorg. Med.l Chem. Lett. 6, 1491-1496; 199 6; Watanabe et al., Tokai J. Experimental Clin. Med. 15, 327-334, 1990; Zhou et al. al., J. Am. Chem. Soc. 126, 15656-7, 2004; WO 01 / 38318; U.S. Patent No. 5,106,951; 5 ,122,368; 5,146,064; 5,177,016; 5,208,323; 5,824,805; 6,146,658; 6,214,345; 7569 358; 7,803,903; 8,084,586; 8,053,205. Antibody-CC-10 via a cross-linker of this patent. Examples of the conjugate structure of 65 analogues are Da01, Da02, Da03, and Da04 below. Examples include a04.

[0211] [ka]

[0212] where mAb is an antibody or cell binding molecule; [ka] represents a single bond or a double bond; n, m1, X1, X2, L1, L2, and R1 are each a group represented by the formula (I ) and (II); R 1’ and R 3’ are each independently, H or C1-C6 linear or branched alkyl; p is 0 to 2000; and further, R 1’ may be Drug 1, a cytotoxic agent described throughout this patent.

[0213] Auristatins and dolastatins ins) are preferred for conjugation via the bridge linkers of the present invention. Auristatins (e.g., Auristatin E (AE), auristatin EB (AEB), auristatin EFP (A EFP), monomethyl auristatin E (MMAE), monomethyl auristatin (MM AF), auristatin F phenylenediamine (AFP), and MMAE phenylalanine Indole-1-4-one (a variant of Indole-1-4-one) is a synthetic analogue of the dolastatin family and is described in the following literature: t. J. Oncol. 15:367-72 (1999); Molecular Cancer Therapeutics, vol. 3, No. 8, pp. 921-932 (2004); U.S. Patent Applications 11134826, 20060074008, 2006022925. U.S. Patent No. 4414 205, 4753894, 4764368, 4816444, 4879278, 4943628, 4978744, 5122368, 5165923, 516 9774,5286637, 5410024, 5521284, 5530097, 5554725, 5585089, 5599902, 5629197, 563 5483, 5654399, 5663149, 5665860, 5708146, 5714586, 5741892, 5767236, 5767237, 57 80588, 5821337, 5840699, 5965537, 6004934, 6033876, 6034065, 6048720, 6054297, 6 054561, 6124431, 6143721, 6162930, 6214345, 6239104, 6323315, 6342219, 6342221, 6407213, 6569834, 6620911, 6639055, 6884869, 6913748, 7090843, 7091186, 7097840, 7098305, 7098308, 7498298, 7375078, 7462352, 7553816, 7659241, 7662387, 7745394 , 7754681, 7829531, 7837980, 7837995, 7902338, 7964566, 7964567, 7851437, 799413 5. An example of the structure of the antibody-auristatin conjugate via the bridge linker of this patent is: The following Au01, Au02, Au03, Au04, Au05, Au06, Au07, Au 08, Au09, Au10, Au11, Au12, and Au13.

[0214] [ka] TIFF0007643734000093.tif179135TIFF0007643734000094.tif161161

[0215] During the ceremony, [ka] n, m1, m2, X1, X2, R1, R2, R3, R4, and R5 are each independently selected from the group consisting of the groups represented by formula (I) and (I I) as defined above; mAb is an antibody or cell-binding molecule; L1, L2, L3, L4, and L5 each independently have the same definition as L1 in formula (I); Z 3’ teeth , H, OPO(O)(OM1)(OM2), OOCCH3, OCH2OP(O)(OM1 ) (OM2), OSO3M1, R1, or O-glycosides (glucosides, galactosides, mannose glycosides, glucuronosides, alosides, fructosides, etc.), NH-glycosides, S-glycosides, or and R1R2, R2R3, R1R3, or R3 R4 is alkyl, aryl, heteroaryl, heteroalkyl, or alkylcycloalky X3 can form a 3- to 8-membered ring of H, CH3, or X 1’ R 1’ and In the formula, X 1’ is NH, N(CH), NHNH, O, or S, and R 1’ is H or C 1-C8 linear or branched alkyl, aryl, heteroaryl, heteroalkyl, alkylcycloalkyl, acyloxyamine; R 3’ is H or a straight chain of C1-C6 or branched alkyl; p is 0 to 2000; M1 and M2 are each independently and H, Na, K, Ca, Mg, NH, NR, R, R; 1’ , Dru g1 and Drug2 are R 1’ The cytotoxic agents described throughout this patent, Dru g1 is also fine.

[0216] Benzodiazepine dimers (e.g., pyrrolobenzoxazoles) are preferred as cytotoxic agents of the present invention. Diazepines (PBD) or (tomaymycin), indolinobenzodiazepines, imidazoline dimers of oxazolidinobenzodiazepines or oxazolidinobenzodiazepines) are known in the prior art. Exemplary in: U.S. Patent Nos. 8,163,736; 8,153,627; 8,034,808; 7,834,005; 7,741 ,319; 7,704,924; 7,691,848; 7,678,787; 7,612,062; 7,608,615; 7,557,099; 7,528,12 8; 7,528,126; 7,511,032; 7,429,658; 7,407,951; 7,326,700; 7,312,210; 7,265,105; 7,202,239; 7,189,710; 7,173,026; 7,109,193; 7,067,511; 7,064,120; 7,056,913; 7,0 49,311; 7,022,699; 7,015,215; 6,979,684; 6,951,853; 6,884,799; 6,800,622; 6,747, 144; 6,660,856; 6,608,192; 6,562,806; 6,977,254; 6,951,853; 6,909,006; 6,344,451 ; 5,880,122; 4,935,362; 4,764,616; 4,761,412; 4,723,007; 4,723,003; 4,683,230; 4 ,663,453; 4,508,647; 4,464,467; 4,427,587; 4,000,304; U.S. Patent Application Nos. 20100203007, 2 0100316656, 20030195196. Conjugation of antibody-benzodiazepine dimers via bridge linkers Examples of body structures are: PB01, PB02, PB03, PB04, PB05, and PB0 6, PB07, PB08, PB09, PB10, and PB11.

[0217] [ka] TIFF0007643734000097.tif215160TIFF0007643734000098.tif79161

[0218] where mAb is an antibody; X3 is CH2, O, NH, NHC(O), NHC(O)N H, C(O), OC(O), OC(O)(NR3), R1, NHR1, NR1, or C(O)R1 or absent; X4 is CH2, C(O), C(O)NH, C( O)N(R1), R1, NHR1, NR1, C(O)R1, or C(O)O; M1 and M2 are each independently H, Na, K, Ca, Mg, NH4, or NR1R2R3. ru; [ka] represents a single bond or a double bond; n, m1, m2, X1, X2, L1, L2, R1, R2, and R3 have the same definition as in formulas (I) and (II); R 1’ and R 3’ are independent H or C1-C6 linear or branched alkyl; p is 0 to 2000; Furthermore, R 1’ may be Drug 1, a cytotoxic agent described throughout this patent.

[0219] Several poisonous mushroom genera, most notably Amanita phalloides (Amanita phalloides) At least 10 toxins found in Mushroom Species (Mushroom spp.) and several other mushroom species A subgroup of compounds, the amatoxins, are also preferred for conjugation via the present bridge linkers. α-amanitin, β-amanitin, γ-amanitin, ε-amanitin, amanulin Ten amanurinic acids, named amanuric acid, amaninamide, amanine, and proamanurin, were Matoxins are rigid bicyclic peptides, synthesized as 35 amino acid proproteins. From there, the last eight amino acids are cleaved off by prolyl oligopeptidase (Litt en, W. 1975 Scientific American232 (3): 90-101;HE Hallen, et al 2007 Proc. N at. Aca. Sci. USA 104, 19097-101; . K. Baumann, et al, 1993 Biochemistry 32 (15) : 4043-50; Karlson-Stiber C, Persson H. 2003, Toxicon 42 (4): 339-49; Horgen, PA et al. 1978 Arch. Microbio. 118 (3): 317-9). Amatoxins bind to RNA polymerases. Potent and selective inhibitors of Pol II, which mediate gene transcription and protein Kills cells by stopping protein biosynthesis (Brodner, OG and Wieland, T. 1976 Biochemistry,15(16): 3480-4; Fiume, L., Curr Probl Clin Biochem, 1977, 7: 23-8; Karlson-Stiber C, Persson H. 2003, Toxicon 42(4): 339-49; Chafin, DR, Guo, H. & Price, DH 1995 J. Biol. Chem. 270 (32): 19114-19; Wieland (1983) Int. J. Pept. Protein Res. 22(3): 257-76.) Amatoxins were detected in collected Amanita faro. From Amanita phalloides mushroom (Yocum, RR 1978 Bioc hemistry 17(18): 3786-9; Zhang, P. et al, 2005, FEMS Microbiol. Lett. 252(2), 22 3-8), or from fermentation using basidiomycetes (Muraoka, S. and Shinozawa T., 2000 J. Biosci. Bioeng. 89(1): 73-6), or from fermentation using A. fissa (Guo, XW, et al, 2 006 Wei Sheng Wu Xue Bao 46(3): 373-8), or Galerina fascicul from cultures of Galerina helvoliceps or Galerina helvoliceps (WO / 1990 / 009799 However, the yields from these isolation and fermentation are very low. During the past 30 years, several amatoxins and The semi-chemical or synthetic preparation of benzophenone and its analogues has been reported (W. E. Savige, A. Fonta). na, Chem. Commun. 1976, 600-1; Zanotti, G., et al, Int J Pept Protein Res, 1981. 18(2): 162-8; Wieland, T., et al, Eur. J. Biochem. 1981, 117, 161-4; PA Bart Lett, et al, Tetrahedron Lett. 1982, 23, 619-22; Zanotti, G., et al., Biochim B iophys Acta, 1986. 870(3): 454-62; Zanotti, G., et al., Int. J. Peptide Protein Res. 1987, 30, 323-9; Zanotti, G., et al., Int. J. Peptide Protein Res. 1987, 30, 450-9; Zanotti, G., et al., Int J Pept Protein Res, 1988. 32(1): 9-20; G. Z anotti, T. et al, Int. J. Peptide Protein Res. 1989, 34, 222-8; Zanotti, G., et al., Int J Pept Protein Res, 1990. 35(3): 263-70; Mullersman, JE and JF Pr eston, 3rd, Int J Pept Protein Res, 1991. 37(6): 544-51; Mullersman, JE, et al , Int J Pept Protein Res, 1991. 38(5): 409-16; Zanotti, G., et al, Int J Pept Pr otein Res, 1992. 40(6): 551-8; Schmitt, W. et al, J. Am. Chem. Soc. 1996, 118, 4380-7; Anderson, MO, et al, J. Org. Chem., 2005, 70(12): 4578-84; JP May, et al, J. Org. Chem. 2005, 70, 8424-30; F. Brueckner, P. Cramer, Nat. Struct. Mol. Biol. 2008, 15, 811-8; JP May, DM Perrin, Chem. Eur. J. 2008, 14, 340 4-9; JP May, et al, Chem. Eur. J. 2008, 14, 3410-17; Q. Wang, et al, Eur. J. Org. Chem. 2002, 834-9; May, JP and DM Perrin, Biopolymers, 2007. 88(5): 7 14-24; May, JP, et al., Chemistry, 2008. 14(11): 3410-7; S. De Lamo Marin, et al, Eur. J. Org. Chem. 2010, 3985-9; Pousse, G., et al., Org Lett, 2010. 12(16) : 3582-5; Luo, H., et al., Chem Biol, 2014. 21(12): 1610-7; Zhao, L., et al., Ch embiochem, 2015. 16(10): 1420-5). Due to their extremely potent and unique cytotoxic mechanism, amatoxins It has been used as a payload for conjugates (Fiume, L., Lancet, 1969. 2 (7625): 853-4 ; Barbanti-Brodano, G. and L. Fiume, Nat New Biol, 1973. 243(130): 281-3; Bonett i, E., M. et al, Arch Toxicol, 1976. 35(1): p. 69-73; Davis, MT, Preston, J. F. Science 1981, 213, 1385-1388; Preston, JF, et al, Arch Biochem Biophys, 19 81. 209(1): 63-71; H. Faulstich, et al, Biochemistry 1981, 20, 6498-504; Barak, LS, et al., Proc Natl Acad Sci USA, 1981. 78(5): 3034-8; Faulstich, H. and L Fiume, Methods Enzymol, 1985. 112: 225-37; Zhelev, Z., A. et al, Toxicon, 1987 . 25(9): 981-7; Khalacheva, K., et al, Eksp Med Morfol, 1990. 29(3): 26-30; U.B. ermbach, H. Faulstich, Biochemistry 1990, 29, 6839-45; Mullersman, JE and J. F. Preston, Int. J. Peptide Protein Res. 1991, 37, 544-51; Mullersman, JE and J. F. Preston, Biochem Cell Biol, 1991. 69(7): 418-27; J. Anderl, H. Echner, H. Faulstich, Beilstein J. Org. Chem. 2012, 8, 2072-84; Moldenhauer, G., et al, J. Natl. Cancer Inst. 2012, 104, 622-34; A. Moshnikova, et al; Biochemistry 2013, 5 2, 1171-8; Zhao, L., et al., Chembiochem, 2015. 16(10): 1420-5; Zhou, B., et al. , Biosens Bioelectron, 2015. 68: 189-96; WO2014 / 043403, US20150218220, EP 166158 4) We have been working on conjugates of amatoxins for some time. Examples of the structure of the antibody-amatoxin conjugate are preferably the following Am01, Am02, Examples include Am03 and Am04.

[0220] [ka] TIFF0007643734000101.tif49155

[0221] where mAb is an antibody; X3 is CH2, O, NH, NHC(O), NHC(O)N H, C(O), OC(O), OC(O)(NR3), R1, NHR1, NR1, or C(O)R1 or absent; R7, R8, R9, R 10 , and R 11 is that each independently H, OH, OR1, NH2, NHR1, or C1-C6 alkyl; Y1 is O, O2, S, or NH or is absent; [ka] represents a single bond or a double bond; n, m1, m2, X1, X2, L1, L2, R1, R2, and R3 have the same definition as in formulas (I) and (II); R 1’ and R 3’ are independent H or C1-C6 linear or branched alkyl; p is 0 to 2000; Furthermore, R 1’ may be Drug 1, a cytotoxic agent described throughout this patent.

[0222] In yet another embodiment, two or more different cytotoxic agents are delivered to cells via a bridge linker of the invention. Preferably, two or more different cytotoxic agents are conjugated to a tubulin-binding molecule. maytansinoids, maytansinoids, taxanoids (taxanes), CC-1065 analogs, Daunorubicin and doxorubicin compounds, benzodiazepine dimers (e.g., pyrrolobe Benzodiazepines (PBDs), tomaymycin, anthramycin, indolinobenzodiazepines azepines, imidazobenzothiadiazepines, or oxazolidinobenzodiazepines dimers), calicheamicin and enediyne antibiotics, actinomycin, amanitin , azaserine, bleomycin, epirubicin, tamoxifen, idarubicin, Dolastatins, auristatins (e.g., monomethyl auristatin E, MMAE, MMAF, auristatin PYE, auristatin TP, auristatin 2-AQ, 6- AQ, EB (AEB), and EFP (AEFP)), duocarmycins, thiotepa, Vincristines, hemiasterins, nazumamides, mica Microginins, radiosumins, Alterobactins, microsclerodermicins (m icrosclerodermins, theonellamides s), esperamicins, PNU-159682, and The bridge linker may be selected from any combination of analogs and derivatives thereof. An example of the structure of a conjugate containing two or more different cytotoxic agents via a hydroxyl group is shown below: Z01, Z02, Z03, Z04, Z05, Z06, Z07, Z08, Z09, Z10, Z11, Z12, Z13, Z14, Z15, Z16, Z17, and Z18.

[0223] [ka] TIFF0007643734000104.tif201166TIFF0007643734000105.tif237163TIFF0007643734000106.tif216161TIFF0007643734000107.tif229163

[0224] wherein mAb is an antibody; X3 and X'3 are each independently CH2, O, NH, NHC(O), NHC(O)NH, C(O), OC(O), OC(O)(NR3), R1 , NHR1, NR1, or C(O)R1, or absent; X4 and X'4 are each independently H, CH2, OH, O, C(O), C(O)NH, C(O)N(R 1), R1, NHR1, NR1, C(O)R1, or C(O)O; M1 and M2 are are independently H, Na, K, Ca, Mg, NH4, NR1R2R3; n, m1, m2, [ka] X1, X2, R1, R2, and R3 have the same definitions as in formulas (I) and (II); R1 and / or R2 may each independently be absent.

[0225] In yet another embodiment, the immunotoxin is conjugated to a cell-binding molecule via a conjugate of the present application. The immunotoxin used herein is generally a diphtheria toxin (DT), a cholera toxin (CTH), or the like. Trichosanthin (CT), trichosanthin (TCS), dianthin, Pseudomonas exotoxin A (ET A'), bacterial or plant proteins such as erythrotoxin, diphtheria toxin, AB toxin, and type III exotoxin It is a macromolecular drug that is a cytotoxic protein derived from proteins. It could be a highly toxic bacterial pore-forming protoxin that requires proteolytic processing for its release. Examples of such protoxins include proaerolysin and its genetically modified forms, and topsalysin. Topsalysin is a prostate A modified recombinant protein designed to be selectively activated by an enzyme, It causes localized cell death and tissue destruction without damaging the underlying tissues and nerves.

[0226] In yet another embodiment, the cell-binding ligand or cell receptor agonist is a conjugate of the present application. These conjugated cell-binding ligands can be conjugated to cell-binding molecules via or receptors, particularly antibody-receptor conjugates, as targeting conductors for delivery of the conjugate to malignant cells. to act as a cytotoxic agent / director, but also to regulate or co-stimulate the desired immune response or can be used to alter signal transduction pathways.

[0227] In immunotherapy, cell-binding ligands or receptor agonists bind to TCR (T cell receptor ) T cells, or CAR (chimeric antigen receptor) T cells, or B cell receptor (BCR), Preferably, the antibody is conjugated to an antibody of a natural killer (NK) cell or a cytotoxic cell. Such antibodies are preferably anti-CD3, CD4, CD8, CD16 (FcγRIII), CD27, CD40, CD40L, CD45RA, CD45RO, CD56, CD57, CD57 bright , TNFβ, Fas ligand, MHC class I molecules (HAL-A, B, C), or NKR-P1. The cell-binding ligand or receptor is not limited to However, folate derivatives (folate receptors, proteins overexpressed in ovarian cancer and other malignancies) (Low, PS et al 2008, Acc. Chem. Res. 41, 120-129); glutamateuria A derivative of thiazolinone (binding to prostate-specific membrane antigen, a surface marker of prostate cancer cells) (Hillier, S. M.et al., 2009, Cancer Res. 69, 6932-6940); somatostatin (growth hormone inhibitor) Mon (GHIH), somatotropin-releasing inhibitory factor (SRIF), or somatotropin-releasing inhibitory hormones) and their derivatives, such as octreotide ( doxorubicin) and lanreotide (somatulin) (especially for neuroendocrine tumors, GH-producing pituitary gland adenoma, paraganglioma, dysfunctional pituitary adenoma, pheochromocytoma) (Ginj, M., et al, 2006, Pro c. Natl. Acad. Sci. USA 103, 16436-16441); in general, somatostatin and its receptors The receptor subtypes (sst1, sst2, sst3, sst4, and sst5) are Many types of tumors, including secretory tumors, especially GH-secreting pituitary adenomas (Reubi JC, Landolt, A. M. 1984 J. Clin. Endocrinol Metab 59: 1148-51; Reubi JC, Landolt AM 1987 J Clin Endocrinol Metab 65: 65-73; Moyse E, et al, J Clin Endocrinol Metab 61: 98-103) and gastroenteropancreatic tumors (Reubi JC, et al, 1987 J Clin Endocrinol Metab 65: 11 27-34; Reubi, J. C, et al, 1990 Cancer Res 50: 5969-77), pheochromocytoma (Epel-baum J, et al 1995 J Clin Endocrinol Metab 80:1837-44; Reubi JC, et al, 1992 J Cl in Endocrinol Metab 74: 1082-9), neuroblastoma (Prevost G, 1996 Neuroendocrinolo gy 63:188-197; Moertel, C. L, et al 1994 Am J Clin Path 102:752-756), thyroid pulp -like cancer (Reubi, J. C, et al 1991 Lab Invest 64:567-573), small cell lung cancer (Sagman U, et al, 1990 Cancer 66:2129-2133), non-metastatic tumors including brain tumors such as meningiomas, medulloblastomas, and gliomas. Transendocrine tumors (Reubi JC, et al 1986 J Clin Endocrinol Metab 63: 433-8; Reubi J C., et al 1987 Cancer Res 47: 5758-64; Fruhwald, M. C, et al 1999 Pediatr Res 45: 697-708), breast cancer (Reubi J. C., et al 1990 Int J Cancer 46: 416-20; Srkalov ic G, et al 1990 J Clin Endocrinol Metab 70: 661-669), lymphoma (Reubi J. C., e t al 1992, Int J Cancer50: 895-900), renal cell carcinoma (Reubi J. C., et al 1992, Cance r Res 52: 6074-6078), mesenchymal tumors (Reubi J. C., et al 1996 Cancer Res 56: 1922-3 1), prostate (Reubi J. C., et al 1995, J. Clin. Endocrinol Metab 80: 2806-14; et al 1989, Prostate 14:191-208; Halmos G, et al J. Clin. Endocrinol Metab 85: 2564 -71), ovary (Halmos, G, et al, 2000 J Clin Endocrinol Metab 85: 3509-12; Reubi J. C., et al 1991 Am J Pathol 138:1267-72), stomach (Reubi J. C., et al 1999, Int J Cancer 81: 376-86; Miller, G. V, 1992 Br J Cancer 66: 391-95), hepatocellular (Kourou malis E, et al 1998 Gut 42: 442-7; Reubi J. C., et al 1999 Gut 45: 66-774), and pharyngeal cancer (Loh K. S, et al, 2002 Virchows Arch 441: 444-8); specific aromatic sulfonamides specific for carbonic anhydrase IX (a marker for hypoxia and renal cell carcinoma) ​Neri, D., et al, Nat. Rev. Drug Discov. 2011, 10, 767-777); Pituitary adenylate cyclase-activating peptide (PACAP) for transglandular tumors 1) for cancers of the lung, stomach, colon, rectum, breast, prostate, pancreatic duct, liver, bladder, and epithelial carcinomas Vasoactive intestinal peptide (VIP) and its receptor subtypes (VPAC1, VCA P2); α-melanocyte-stimulating hormone (α-MSH) receptors for various tumors; small cell lung Cholecystokinin for cancer, medullary thyroid carcinoma, astrocytoma, insulinoma, and ovarian cancer (CCK) / gastrin receptors and their receptor subtypes (CCK1 (formerly CCK -A) and CCK2; renal cell carcinoma, breast cancer, lung cancer, gastric cancer, and prostate cancer, as well as neuroblastoma ( and bombesin (Pyr-Gln-Arg-Leu-Gly-Asn-Gln-) for neuroblastoma (neuroblastoma). Trp-Ala-Val-Gly-H is-Leu-Met-NH2) / gastrin-releasing peptide (GRP) and its receptor Body subtypes (BB1, GRP receptor subtype (BB2), BB3, and BB4) (Oh lsson, B., et al, 1999, Scand. J. Gastroenterology 34 (12): 1224-9; Weber, H. C ., 2009, Cur. Opin. Endocri. Diab. Obesity 16(1): 66-71, Gonzalez N, et al, 2008 , Cur. Opin. Endocri. Diab. Obesity 15(1), 58-64); small cell lung cancer, neuroblastoma, pancreatic Neurotensin (NTR1, NTR2, NTR3) for pancreatic and colon cancer; glial Substance P and its receptor subtypes in tumors (e.g., NK1 receptor for glial tumors, Hennig IM, et al 1995 Int. J. Cancer 61, 786-792); Neuropeptides for breast cancer Y thiol and its receptor subtypes (Y1-Y6); RGD (Arg-Gly-Asp), NGR (Asn -Gly-Arg), a dimeric and multimeric cyclic RG that recognizes receptors (integrins) on tumor surfaces D peptide (e.g. cRGDfV) (Laakkonen P, Vuorinen K. 2010, Integr Biol ( Camb). 2(7-8): 326-337; Chen K, Chen X. 2011, Theranostics. 1:189-200; Garanger E, et al, Anti-Cancer Agents Med Chem. 7 (5): 552-558; Kerr, JS et al, Antica Ncer Research, 19(2A), 959-968; Thumshirn, G, et al, 2003 Chem. Eur. J. 9, 2717- 2725), and TAASGVRSMH and LTLRWVGLMS (chondroitin sulfate proteoglycan NG2 receptor) and F3 peptide (cell surface-expressed nucleolin receptor) 31 amino acid peptide that binds to the IL-1 receptor (Zitzmann, S., 2002 Cancer Res., 62, 18, pp. 5139 -5143, Temminga, K., 2005, Drug Resistance Updates, 8, 381-402; P. Laakkonen an d K. Vuorinen, 2010 Integrative Biol, 2(7-8), 326-337; MA Burg, 1999 Cancer R es., 59(12), 2869-2874; K. Porkka, et al 2002, Proc. Nat. Acad. Sci. USA 99(11) , 7444-9); cell-penetrating peptides (CPPs) (Nakase I, et al. l, 2012, J. Control Release. 159(2),181-188); peptide hormones, e.g., testosterone As well as steroid production, it also regulates the production of follicle-stimulating hormone (FSH) and luteinizing hormone (LH). Luteinizing hormone-releasing hormone (LHRH) agonists and antagonists that act by targeting agonists, as well as gonadotropin-releasing hormone (GnRH) agonists, such as Buserene Phosphorus (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-Pro-NHEt), gonadorelin (Pyr-His- Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2, goserelin (Pyr-His-Trp-Ser-Tyr-D-Ser(OtB u)-Leu-Arg-Pro-AzGly-NH2), histrelin (Pyr-His-Trp-Ser-Tyr-D-His(N-benzyl)- Leu-Arg-Pro-NHEt), leuprolide (Pyr-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro-NHEt), Nafarelin (Pyr-His-Trp-Ser-Tyr-2Nal-Leu-Arg-Pro-Gly-NH2), Triptorelin (Py r-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH2), nafarelin, deslorelin, ava Relics (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-(N-Me)Tyr-D-Asn-Leu- isopropylLys-Pro-DAla-NH2), Cetrorelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl) Lysyl)Ala-Ser-Tyr-D-Cit-Leu-Arg-Pro-D-Ala-NH2), degarelix (Ac-D-2Nal-D-4- chloroPhe-D-3-(3-pyridyl)Ala-Ser-4-aminoPhe(L-hydroorotyl)-D-4-aminoPhe(calcium phosphate) bamoyl)-Leu-isopropylLys-Pro-D-Ala-NH2), and ganirelix (Ac-D-2Nal-D-4-ch loroPhe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-(N9,N10-diethyl)-homoArg-Leu-(N9,N10-diethyl) Ethyl)-homoArg-Pro-D-Ala-NH2)(Thundimadathil, J., J. Amino Acids, 2012, 96734 7, doi:10.1155 / 2012 / 967347; Boccon-Gibod, L.; et al, 2011, Therapeutic Advances in Urology 3 (3): 127-140; Debruyne, F., 2006, Future Oncology, 2(6), 677-696; Schally A. V; Nagy, A. 1999 Eur J Endocrinol 141:1-14; Koppan M, et al 1999 Pros tate 38:151-158); and small molecules such as imiquimod, guanidine, and adenosine Lipopolysaccharide (LPS), nucleic acids (CpG DNA, poly I;C) and lipopeptides Pam3CSK4 (Kasturi, SP, et al, 2011, Nature 470, 543-547; Lan e, T., 2001, JR Soc. Med. 94, 316; Hotz, C., and Bourquin, C., 2012, Oncoimmu 1, 227-228; Dudek, AZ, et al, 2007, Clin. Cancer Res. 13, 7119-7125) Toll-like receptors (TLRs), which recognize large and complex biological macromolecules such as C type lectins, and Nod-like receptors (NLRs) (Fukata, M., et al., 2009, Semin. Immu nol. 21, 242-253; Maisonneuve, C., et al, 2014, Proc. Natl. Acad. Sci. USA 111, 1-6; Botos, I., et al, 2011, Structure 19, 447-459; Means, TK, et al, 2 000, Life Sci. 68, 241-258) and other pattern recognition receptors (PRRs). A 32-amino acid neuropeptide that is broadly involved in regulating calcium levels through its effects on Calcitonin receptor (Zaidi M, et al, 1990 Crit Rev Clin Lab Sci 28, 109-174 ; Gorn, AH, et al 1995 J Clin Invest 95:2680-91); various cells, especially osteoclasts, It is expressed on the surface of endothelial cells and tumor cells and generally plays an important role in angiogenesis. Integrin receptors and their receptor subtypes (e.g., αVβ1, αVβ3, αV β5, αVβ6, α6β4, α7β1, αLβ2, αIIbβ3) (Ruoslahti, E. et a l, 1994 Cell 77, 477-8; Albelda, SM et al, 1990 Cancer Res., 50, 6757-64). A short peptide and GRGDSPK that showed high binding affinity to integrin receptors, and Cyclic RGD pentapeptides such as cyclo(RGDfV)(L1) and their derivatives [cyclo( -N(Me)R-GDfV), cyclo(R-Sar-DfV), cyclo-(RG-N(M e)D-fV), cyclo(RGD-N(Me)fV), cyclo(RGDf-N(Me) V-)(Cilengitide)](Dechantsreiter, MA et al, 1999 J. Med. Ch em. 42, 3033-40, Goodman, SL, et al, 2002 J. Med. Chem. 45, 1045-51).

[0228] Cell-binding ligands or cell receptor agonists bind to Ig-based and non-Ig-based proteins. The Ig-based scaffold can be, but is not limited to, Nanobodies (VHH (camelid Ig) derivatives) (Muyldermans S., 20 13 Annu Rev Biochem. 82, 775-797); domain antibodies (dAb, VH or VL domains) derivatives) (Holt, L. J, et al, 2003, Trends Biotechnol. 21, 484-490); bispecific T cell induction (BiTE, bispecific dimer) (Baeuerle, P. A, et al, 2009, Curr. Opin. M ol. Ther. 11, 22-30); dual affinity retargeting (DART, bispecific dimer) (Moore P A. P, et al. 2011, Blood 117(17), 4542-4551; tetravalent tandem antibody (TandAb , bispecific dimer) (Cochlovius, B, et al. 2000, Cancer Res. 60(16):4336-4341) Non-Ig scaffolds can be selected from, but are not limited to, anticalins ( Lipocalin derivatives) (Skerra A. 2008, FEBS J., 275(11): 2677-2683; Beste G, et al, 1999 Proc. Nat. Acad. USA. 96(5):1898-1903; Skerra, A. 2000 Biochim Biophys Acta, 1482(1-2):337-350; Skerra, A. 2007, Curr Opin Biotechnol. 18(4):295-304; Skerra, A. 2008, FEBS J. 275(11):2677-2683; Adnectins (FN3 (Finally bronectin)) (Koide, A, et al, 1998 J. Mol. Biol., 284(4):1141-1151; Batori V, 2002, Protein Eng. 15(12): 1015-1020; Tolcher, A. W, 2011, Clin. Cancer Re s. 17(2):363-371; Hackel, B. J, 2010, Protein Eng. Des. Sel. 23(4):211-219); Developed ankyrin repeat proteins (DARPins) (ankyrin repeat (AR) ) Protein derivatives) (Boersma, YL, et al, 2011 Curr Opin Biotechnol. 22(6): 8 49-857), such as DARPinC9, DARPinEc4, and DARPinE69_L Z3_E01(Winkler J, et al, 2009 Mol Cancer Ther. 8(9), 2674-2683; Patricia MK. M., et al, Clin Cancer Res. 2011;17(1):100-110; Boersma Y. L, et al, 2011 J Biol. Chem. 286(48),41273-41285; Avimer (Domain A / Low Density Low-density lipoprotein (LDL) receptor) (Boersma Y. L, 2011 J. Biol. Chem. 286(48): 4 1273-41285; Silverman J, et al, 2005 Nat. Biotechnol., 23(12):1556-1561). You can choose.

[0229] Structure of the antibody-cell binding ligand or cell receptor agonist conjugate via the present linker Examples are: LB01 (folic acid conjugate), LB02 (PMSA ligand conjugate) ), LB03 (PMSA ligand conjugate), LB04 (somatostatin conjugate), LB0 5 (octreotide, somatostatin analogue conjugate), LB06 (lanreotide, somatostatin Statin analogue conjugate), LB07 (vapreotide (Sanvar), somatostatin analogue Ligand conjugate), LB08 (CAIX ligand conjugate), LB09 (CAIX ligand conjugate) LB10 (gastrin-releasing peptide receptor (GRPr), MBA conjugate), LB1 1 (luteinizing hormone-releasing hormone (LH-RH) ligand and GnRH conjugate), LB 12 (luteinizing hormone-releasing hormone (LH-RH) and GnRH ligand conjugate), L B13 (GnRH antagonist, abarelix conjugate), LB14 (cobalamin, V B12 analogue conjugate), LB15 (cobalamin, VB12 analogue conjugate), LB16 (α v β3 integrin receptor, cyclic RGD pentapeptide conjugate), LB17 (VEGF receptor Heterobivalent peptide-ligand conjugate for receptors), LB18 (Neuromedin B conjugate) ), LB19 (bombesin conjugate for G protein-coupled receptors), and LB20 (T TLR2 conjugate for IgE-like receptor), LB21 (for androgen receptor), LB22(α V Cilengitide / cyclo(-RGDfV)- for integrin receptors ) conjugate), LB23 (fludrocortisone conjugate), LB24 (dexamethasone conjugate) ), LB25 (fluticasone propionate conjugate), LB26 (beclomethasone dipropionate conjugate) LB27 (triamcinolone acetonide conjugate), LB28 (prednisone conjugate) LB29 (prednisolone conjugate), LB30 (methylprednisolone conjugate) , LB31 (betamethasone conjugate), LB32 (irinotecan analogue), LB33 (cristine) Zotinib analogue), LB34 (bortezomib analogue), LB35 (carfilzomib analogue) LB36 (carfilzomib analogue), LB37 (leuprolide analogue), LB3 8 (triptorelin analogue), LB39 (liraglutide analogue), LB40 (semaglutide LB41 (lixisenatide analog), and LB41 (lixisenatide analog). The structures of these are shown below:

[0230] [ka] TIFF0007643734000110.tif252151TIFF0007643734000111.tif251153TIFF0007643734000112.tif253157TIFF0007643734 000113.tif240170TIFF0007643734000114.tif218167TIFF0007643734000115.tif232165TIFF0007643734000116.tif25016 9TIFF0007643734000117.tif249153TIFF0007643734000118.tif240164TIFF0007643734000119.tif241145TIFF0007643734 000120.tif251158TIFF0007643734000121.tif249150TIFF0007643734000122.tif233167TIFF0007643734000123.tif76169

[0231] where mAb is an antibody; X3 is CH2, O, NH, NHC(O), NHC(O)N H, C(O), OC(O), OC(O)(NR3), R1, NHR1, NR1, or C(O)R1 or absent; X4 is H, CH2, OH, O, C(O), C( O)NH, C(O)N(R), R, NHR, NR, C(O)R, or C(O) X5 is H, CH3, F, or Cl; M1 and M2 are each independently H, Na, K, Ca, Mg, NH, NR1R2R3; R6 is 5'-deoxyadenosine Nosyl, Me, OH, or CN; [ka] represents a single or double bond; m1, m2, n, [ka] X1, X2, R1, and R2 have the same definitions as in formula (I); furthermore, R1 may be absent. and R2 may be hydrogen.

[0232] In yet another embodiment, one, two, or more of DNA, RNA, mRNA, small molecule interferon, Interfering RNA (siRNA), microRNA (miRNA), and PIWI-interacting RNA (piRNA) is preferably conjugated to a cell-binding molecule via the conjugate of the present application. (siRNA, miRNA, piRNA) and long non-coding antisense RNA , which are known to be involved in epigenetic changes within cells (Goodchild, J (2011), Methods in molecular biology (Clifton, NJ). 764: 1-15). DNA, RNA, mRNA, siRNA, miRNA, or piRNA in It may be single-stranded or double-stranded, with up to 1 million nucleotide units, Some of the drugs are phosphorothioates, exemplified by fomivirsen. Oate-linked oligonucleotides or mipomersen ), which uses phosphorothioate bonds rather than the phosphodiester bonds of natural RNA and DNA. They are linked by a thioate bond, and the sugar moiety in the center of the molecule is deoxyribose, with two A nucleotide or peptide that is 2'-O-methoxyethyl modified ribose at the end Nucleic acid (PNA), morpholino, phosphorothioate, thiophosphoramidate, or 2' -O-Methoxyethyl (MOE), 2'-O-methyl, 2'-fluoro, locked nucleic acid ( oligonucleotides made with bicyclic nucleic acids (LNA) or ribose sugars or nucleic acids modified to remove the 2'-3' carbon bond in the sugar ring structure. It may also be in a non-natural (synthetic) form, such as (Whitehead, KA; et al (2011), Annual Review w of Chemical and Biomolecular Engineering 2: 77-96; Bennett, CF; Swayze, EE (2010), Annu. Rev. Pharmacol. Toxicol. 50: 259-29). Preferably, oligonucleotides The length of the conjugate ranges from about 8 to over 100 nucleotides. Examples of conjugate structures are shown below: show:

[0233] [ka] TIFF0007643734000127.tif54160

[0234] In the formula, mAb, m1, m2, n, X1, X2, X3, X4, R 1’ , R 2’ , L1, L 2, L3, L4, [ka] is the same as in formula (I) or above; [ka] is a single strand of DNA, RNA, mRNA, siRNA, miRNA, or piRNA It is double-stranded; X5 has the same definition as X1; Y and Y' are O, S, NH, or CH2 is.

[0235] an IgG antibody, a monoclonal antibody, or an IgG antibody-like protein; The conjugate may be an IgG antibody conjugated to one, two, or more of the same or different functional molecules or drugs. The conjugate preferably has the following structure: ST1, ST2, ST3, ST4, ST5, ST 6, ST7, ST8, ST9, ST10, ST11, ST12, ST13, or ST14 As shown in Figure 1, a pair of thiols (due to reduction of disulfide bonds) are formed between the light and heavy chains. The upper disulfide bond between one heavy chain and the lower disulfide bond between the two heavy chains is specifically conjugated to

[0236] [ka] JPEG0007643734000131.jpg141114

[0237] In the formula, X1, X 1’ , X2, X 2’ , X3, X 3’ , X4, X 4’ , L1, L 1’ , L 2. L 2’ , L3, L 3’ , L4, L 4’ and T has the same definition as X1 in formula (I). Furthermore, X1, X 1’ , X2, X 2’ , X3, X 3’ , X4, X 4’ may not exist .

[0238] In yet another embodiment, a therapeutically effective amount of a conjugate of formula (II) or any of the compounds described throughout this patent is administered. Pharmaceutical compositions containing any of the conjugates may be used in chemotherapy, radiation therapy, immunotherapy, autoimmune disease, and the like. and other conjugates such as anti-cancer agents, anti-infective agents, or other conjugates for the synergistically effective treatment or prevention of cancer. The synergistic agent may be administered simultaneously with the therapeutic agent. Preferably, the drug is selected from the following: abatacept (Orencia), abiraterone acetate (Zytig) a®), acetaminophen / hydrocodone, aducanumab, adalimumab, ADXS31-142 (Advaxis'), ADXS-HER2 (Advaxis'), Afatinib dimaleate (Gilotrif®), alemtuzumab (Campath®), Alitretionin (Panretin®), ado-trastuzumab (Kadcyla®) amphetamine mixed salts (amphetamine / dextroamphetamine, or added ral XR), anastrozole (Arimidex®), aripiprazole, and atazana Bill, atezolizumab (MPDL3280A), atorvastatin, axitinib (Inlyta (registered) trademark), avelumab, belinostat (Beleodaq™), bevacizumab (Avast in®), cabazitaxel (Jevtana®), cabozatinib (Cometriq®), (trademark), bexarotene (Targretin®), blinatumomab (Blincyto®), bortezomib (Velcade®), bosutinib (Bosulif®) , brentuximab vedotin (Adcetris®), budesonide, budesonide / hormones Moterol, buprenorphine, capecitabine, carfilzomib (Kyprolis® )), celecoxib, ceritinib (LDK378 / Zykadia), cetuximab (Erbitux (registered trademark) (trademark), cyclosporine, cinacalcet, crizotinib (Xalkori®), Cosentyx, Kymriah, dabigatran, dabrafenib finlar®), daratumab (Darzalex), darbepoetin alfa, darunavir, Imatinib (Gleevec®), Dasatinib (Sprycel®), Denalifloxacin (Denalifloxacin), Leukin diftitox (Ontak®), denosumab (Xgeva®), Depakote, dexlansoprazole, dexmethylphenidate, dexamethasone Zon, Dignitana DigniCap Cooling System, Jinuts Ximab (Unituxin™), doxycycline, duloxetine, duvelisib, Lotuzumab, emtricitabine / rilpivirine / tenofovir disoproxil fumarate, Mutricitabine / tenofovir / efavirenz, enoxaparin, enzalutamide (Xtan di(R), epoetin alfa, erlotinib (Tarceva(R)), esomeprazole zole, eszopiclone, etanercept, everolimus (Afinitor®), Xemestane (Aromasin®), everolimus (Afinitor®), eze Ezetimibe, ezetimibe / simvastatin, fenofibrate, filgrastim, fin Golimod, fluticasone propionate, fluticasone / salmeterol, fulvestran Faslodex®, Gazyva, gefitinib (Iressa®), (label), glatiramer, goserelin acetate (Zoladex), icotinib, imatinib (Gleevec ), ibritumomab tiuxetan (Zevalin®), ibrutinib (Imbruvica (trademark), idelalisib (Zydelig®), infliximab, iniparib, Insulin aspart, insulin detemir, insulin glargine, insulin lysine Pro, interferon beta 1a, interferon beta 1b, lapatinib (Tykerb (registered trademark) ipilimumab (Yervoy®), ipratropium bromide / salbutamin Lu, Ikusazomi (Ninlaro®), Kanuma, lanreotide acetate (Somatuline® depot), lenalidomide, lenalidomide (Revlimid® lenvatinib mesylate (Lenvima™), letrozole (Femara® )), levothyroxine, lidocaine, linezolid, liraglutide, lis Dexamfetamine, LN-144 (Lion Biotech), MEDI4736 (AstraZeneca, Celgene), memantine, methylphenidate, metoprolol, Mekinist, Modafi Nil, mometasone, nilotinib (Tasigna®), niraparib, nivolumab (Opd ivo®), ofatumumab (Arzerra®), obinutuzumab (Gazyva®), (TM), olaparib (Lynparza™), olmesartan, olmesartan / hydrazide Lochlorothiazide, omalizumab, omega-3 fatty acid ethyl esters, oseltamivir, oxycodone Codon, palbociclib (Ibrance®), palivizumab, panitumumab (Vecti bix®), panobinostat (Farydak®), pazopanib (Votrient (R), pembrolizumab (Keytruda®), pemetrexed (Alimta ), pertuzumab (Perjeta™), pneumococcal conjugate vaccine, pomalidomide (Pomalys t(R)), pregabalin, ProscaVax, propranolol, quetiapine rabeprazole, radium-223 chloride (Xofigo®), raloxifene , raltegravir, ramucirumab (Cyramza®), ranibizumab, regorafenib Nib (Stivarga®), rituximab (Rituxan®), rivaroxaban romidepsin (Istodax®), rosuvastatin, ruxolitinib phosphate (J akafi™), salbutamol, sevelamer, sildenafil, siltuximab (Syl vant™), sitagliptin, sitagliptin / metformin, solifenacin, ranezumab, sorafenib (Nexavar®), sunitinib (Sutent®) ), tadalafil, tamoxifen, Tafinlar, talazoparib , telaprevir, temsirolimus (Torisel®), tenofovir / emtricitabine bottle, testosterone gel, thalidomide (Immunoprin, Talidex), tiotropium bromide toremifene (Fareston®), trametinib (Mekinist®), Trastuzumab, tretinoin (Vesanoid®), ustekinumab, valsal vemurafenib (Zelboraf®), vandetanib (Caprelsa®), veliparib (Vandetanib), (registered trademark), venetoclax, vorinostat (Zolinza®), Zivaf Libercept (Zaltrap®), and Zostavax, and their analogs; derivatives, pharmaceutically acceptable salts thereof, carriers, diluents or excipients therefor, or A combination of these.

[0239] The drugs / cytotoxic agents used in the conjugation via the bridge linkers of this patent are The compound may be any analogue and / or derivative of the drug / molecule. Those skilled in the art will recognize that the resulting compound still retains the specificity and / or activity of the starting compound. It is readily apparent that each of the drugs / cytotoxic agents described herein can be modified to Those skilled in the art will also appreciate that many of these compounds are also compounds that are suitable for use with the drugs / cell injury compounds described herein. It will be understood that the drug / cytotoxic agent of the present invention may be used in place of the cytotoxic agent. Agents include analogs and derivatives of the compounds described herein.

[0240] All references cited in the specification and examples that follow are expressly incorporated by reference in their entirety. It can be enjoyed. [Example]

[0241] The present invention is further illustrated by the following examples, which should not be construed as limiting the scope of the present invention. The cell lines described in the following examples are not intended to be representative unless otherwise specified. American Type Culture Collection (ATCC), German Collection of Microbial Cell Cultures Based on the conditions specified by the Shanghai Institute of Cell Culture, Chinese Academy of Sciences (DSMZ) Unless otherwise stated, all cell culture reagents were purchased from Invitrogen. All anhydrous solvents were obtained commercially and stored in nitrogen-filled sealed bottles. All other reagents and solvents were purchased according to the highest standards and used without further purification. Preparative HPLC separations were performed on a Varain PreStar HPLC. R spectra were detected on a Varian Mercury 400 MHz instrument. The chemical shift (Δ) is in ppm, and the bond constant is 0.00 relative to tetramethylsilane. The unit of (J) is Hz. Mass spectrometry data was obtained from Waters Acquity UPL Waters Xevo QTO equipped with a C separator and Acquity TUV detector F mass spectrometry.

[0242] Example 1: 1,2-bis(2-(tert-butoxy)-2-oxoethyl)hydrazine Synthesis of di-tert-butyl 1,2-dicarboxylate (38) [ka]

[0243] Di-tert-butyl hydrazine-1,2-dicarboxylate ( 37) (8.01 g, 34.4 mmol) to NaH (60% in oil, 2.76 g, 68.8 After stirring at room temperature for 30 minutes, tert-butyl 2-bromoacetate (1 mmol) was added. The mixture was stirred overnight and then diluted with methanol (3 ml). Quench by addition of, concentrate, and dilute with EtOAc (100 mL) and water (100 mL). The organic layers were combined and separated, and the aqueous layer was extracted with EtOAc (2 x 50 mL). Drying with gSO4, filtering, evaporating, and eluting with SiO2 column chromatography (Et Purification with OAc / hexane 1:5 to 1:3 gave the title compound (12.98 g, yield 8 2%) as a colorless oil. MS ESI m / z calculated C 22 H 41 N2O8[M+H] + 461.28, measured value 461.40.

[0244] Example 2: Synthesis of 2,2'-(hydrazine-1,2-diyl)diacetic acid (39) [ka]

[0245] 1,2-bis(2-(tert-butoxy)-2-methylpropional) in 1,4-dioxane (40 ml) -oxoethyl)hydrazine-1,2-dicarboxylate di-tert-butyl (6.51g , 14.14 mmol) was added HCl (12 M, 10 ml). The mixture was stirred for 30 min. Diluted with dioxane (20 ml) and toluene (40 ml), evaporated, and distilled off. The mixture was evaporated to dryness with xanthene (20 ml) and toluene (40 ml) without further preparation. The crude product of the title compound for the next step was obtained (2.15 g, 103% yield, approximately 93% purity). %). MS ESI m / z calculated value C4H9N2O4[M+H] + 149.05, measured value 149.40.

[0246] Example 3: 2,2'-(1,2-bis((E)-3-bromoacryloyl)hydrazine- Synthesis of 1,2-diyl)diacetic acid (36) [ka]

[0247] A mixture of THF (50 ml) and NaH2PO4 (0.1 M, 80 ml, pH 6.0) 2,2'-(hydrazine-1,2-diyl)diacetic acid (1.10 g, 7.43 mmol) ) solution, (E)-3-bromoacryloyl bromide (5.01 g, 23.60 mmol) l) was added. The mixture was stirred for 6 h, concentrated, and diluted with H2O / CH3Cl containing 3% formic acid. Purification on a SiO2 column eluted with N (1:9) gave the title compound (2.35 g, yield 77%, purity approximately 93%). MS ESI m / z calculation value C 10 H 11 Br2N2O6[M+H] + 412.89, measured 413. 50.

[0248] Example 4: 2,2'-(1,2-bis((E)-3-bromoacryloyl)hydrazine- Synthesis of 1,2-diyl)diacetyl chloride (41) [ka]

[0249] 2,2'-(1,2-bis((E)-3-bromoacetic acid) in dichloroethane (15 mL) (1,2-dihydroxybenzoyl)hydrazine-1,2-diyl)diacetic acid (210 mg, 0.509 mmol) , (COCl)2 (505 mg, 4.01 mmol) was added, followed by 0.040 ml of DMF was added. After stirring at room temperature for 2 hours, the mixture was concentrated and diluted with dichloroethane (2×2 It was evaporated to dryness with toluene (2 x 10 mL) and toluene (2 x 15 mL) for further purification. Without further elaboration, the crude title product (not stable) for the next step was obtained (245 mg, yield 107 %). MS ESI m / z calculation value C 10 H9Br2Cl2N2O4[M+H] + 448.82, 450.82, 452.82, 454.82, Measurement Values ​​448.60, 450.60, 452.60, 454.60.

[0250] Example 5: tert-Butyl 2,8-dioxo-1,5-oxazocane-5-carboxylate Synthesis of (47) [ka]

[0251] 3,3'-Azanediyldipropanoic acid (42) (10.00 g, 62.08 mmol) in 1.0 M NaOH (300 mL) A solution of dicarbonate (22.10 g, 101.3 mmol) in THF (200 mL) was added. After the addition, the mixture was stirred for 2 hours at 4°C. The mixture was diluted with 0.2M H3PO4 Carefully acidify to pH 4 with HCl, concentrate in vacuo, dry with CH2Cl2, and evaporate. Flash SiO2 eluted with AcOH / MeOH / CH2Cl2 (0.01:1:5) Purified by chromatography to give 3,3'-((tert-butoxycarbonyl)azanediol). (46) (13.62 g, 84% yield). ESI MS m / z calculated C 11 H 19 NO6[M+H] + 262.27, measured value 262.40.

[0252] At 0°C, 3,3'-((tert-butoxycarbonyl)azanediyl)dipropanoic acid (8.0 g, 30.6 mmol) in CHCl (500 mL) The mixture was stirred at 0°C for 2 hours and then at room temperature for 1 hour. The mixture was stirred for 10 minutes, filtered through a short SiO2 column, and then the column was filled with EtOAc / CH2Cl2 (1:6). The filtrate was concentrated and triturated with EtOAc / hexane to give the title compound ( 47) was obtained (5.64 g, 74% yield). ESI MS m / z calculated C 11 H 17 NO5[M+H] + , 244.1 1, Measurement value 244.30.

[0253] Example 6: Synthesis of 2,5-dioxopyrrolidin-1-ylpropiolate (61) [ka]

[0254] Propionic acid (5.00 g, 71.4 mmol), NHS (9.01 g, 78.3 mmol) ol), and EDC (20.0 g, 104.1 mmol) in CH2Cl2 (150 ml L) and DIPEA (5 mL, 28.7 mmol) were stirred overnight and evaporated to give Si Purification by O2 chromatography (EtOAc / hexane 1:4) afforded the title compound as a colorless Obtained as an oil (9.30 g, 79% yield). 1 H NMR (500 MHz, CDCl3) δ 2.68 (s, 1H), 2.61 (s, 4H). MS ESI m / z calculated value C7H5NaNO4[M+Na] + 190.02, measured value 190.20.

[0255] Example 7: Synthesis of tert-butyl 2-propioloylhydrazinecarboxylate (88) [ka]

[0256] Propionic acid (5.00 g, 71.4 mmol), tert-butyl hydrazine carboxylate Chill (9.45 g, 71.5 mmol), and EDC (20.0 g, 104.1 mmol) ) in CH2Cl2 (150 mL), and DIPEA (5 mL, 28.7 mmol) Stir overnight, evaporate and chromatograph on SiO2 (EtOAc / hexane 1:5 ) to give the title compound as a colorless oil (7.92 g, 84% yield). 1 H NMR (500 MHz, CDCl3) δ 8.76 (m, 2H), 2.68 (s, 1H), 1.39 (s, 9H). MS ESI m / z calculated value C5H 12 Na N2O2[M+Na] + 155.09, measured value 155.26.

[0257] Example 8: Synthesis of propiolohydrazide hydrochloride (89) [ka]

[0258] 2-Propioloylhydrazinecarboxylic acid dissolved in 1,4-dioxane (12 mL) tert-Butyl ester (4.01 g, 30.35 mmol) was treated with 4 mL of concentrated HCl. The mixture was stirred for 30 minutes and diluted with dioxane (30 mL) and toluene (30 mL). The crude mixture was concentrated in vacuo and diluted with methanol (5%-10%) and 1% formic acid in methylene chloride. The mixture of solutions was purified on silica gel to give the title compound (2.11 g, 8% yield). 3%). ESI MS m / z calculated value C3H5N2O [M+H] + , 85.03, Measurement 85.30.

[0259] Example 9: (S,E)-2-methyl-N-(3-methylbutan-2-ylidene)propane Synthesis of 2-sulfonamide (186) [ka]

[0260] (S)-2-methylpropane-2-sulfinamide (100 g, 0.825 mol, 1.0 equiv.) in THF (1 L), Ti(OEt)4 (345 mL, 1.82 mol , 2.2 equiv.) and 3-methyl-2-butanone (81 mL, 0.825 mol, 1.0 equiv.) The reaction mixture was refluxed at room temperature under N2 for 16 hours, then cooled to room temperature and poured into ice water. The mixture was filtered and the filter cake was washed with EtOAc. The organic layer was separated. The residue was then dried over anhydrous Na2SO4 and concentrated to give a residue, which was then distilled under vacuum (15-20 torr). r, 95°C) to give the title compound as a yellow oil (141 g, 90% yield) . 1 H NMR (500 MHz, CDCl3) δ 2.54 - 2.44 (m, 1H), 2.25 (s, 3H), 1.17 (s, 9H), 1. 06 (dd, J = 6.9, 5.1 Hz, 6H). MS ESI m / z calculated value C9H 19 NaNOS [M+Na] + 212.12; Measurement 2 12.11.

[0261] Example 10: Synthesis of (2S,3S)-2-azido-3-methylpentanoic acid (177) [ka]

[0262] NaN3 (20 mL) in a mixture of water (50 mL) and dichloromethane (80 mL) at 0 °C. To a solution of TfO (10 mL, 59.2 mmol, 2.0 eq. After the addition, the mixture was stirred at 0°C for 2 hours, and the organic phase was separated. The organic phase was washed with saturated NaHCO3 solution and A solution of triflurazide in dichloromethane was dissolved in water (100 mL) and methanol ( (L)-Isoleucine (4.04 g, 30.8 mmol, 1.0 equiv.) in 200 mL of water , K2CO3 (6.39 g, 46.2 mmol, 1.5 equiv.), and CuSO4 5H2 The mixture was added with 200 mL of HCl (77.4 mg, 0.31 mmol, 0.01 equiv.). The organic solvent was reduced. The aqueous phase was removed under pressure, diluted with water (250 mL), acidified to pH 6 with concentrated HCl, and phosphoric acid The aqueous phase was diluted with 250 mL of 0.25 M buffer (pH 6.2). 00 mL) to remove the sulfonamide by-product, then acidify to pH 2 with concentrated HCl The combined organic layers were dried over anhydrous Na2SO4. The mixture was evaporated, filtered, and concentrated to give the title compound as a colorless oil (4.90 g, 99% yield). %). 1 H NMR (500 MHz, CDCl3) δ 12.01 (s, 1H), 3.82 (d, J = 5.9 Hz, 1H), 2.00 ( ddd, J = 10.6, 8.6, 5.5 Hz, 1H), 1.54 (dqd, J = 14.8, 7.5, 4.4 Hz, 1H), 1.36 - 1 .24 (m, 1H), 1.08 - 0.99 (m, 3H), 0.97 - 0.87 (m, 3H).

[0263] Example 11: Synthesis of DN-methylpipecolic acid [ka]

[0264] D-pipecolic acid (10.0 g, 77.4 mmol, 1.0 equiv.) in methanol ( 100 mL) of formaldehyde (37% aqueous solution, 30.8 mL, 154.8 mmol) , 2.0 equiv.), followed by Pd / C (10 wt%, 1.0 g). The reaction mixture was stirred overnight. Stir under H2 (1 atm), filter through Celite, then rinse the filter pad with methanol. The filtrate was concentrated under reduced pressure to give the title compound as a white solid (10.0 g , yield 90%).

[0265] Example 12: (R)-Perfluorophenyl 1-methylpiperidine-2-carboxylate Synthesis of acetone [ka]

[0266] DN-methylpipecolic acid (2.65 g, 18.5 mmol) in EtOAc (5 0 mL), pentafluorophenol (3.75 g, 20.4 mmol) and DCC ( The reaction mixture was stirred at room temperature for 16 hours, and then The mixture was filtered over Celite. The filter pad was washed with 10 mL of EtOAc. The filtrate was and used immediately without further purification or concentration.

[0267] Example 13: Synthesis of 2,2-diethoxyethanothioamide (180) [ka]

[0268] At room temperature, 2,2-diethoxyacetonitrile (100 g, 0.774 mol, 1.0 equivalents) (amount) and (NH4)2S aqueous solution (48%, 143 mL, 1.05 mol, 1.36 equiv.) and were mixed in methanol (1.5 L). After stirring for 16 hours, the reaction mixture was concentrated and The residue was taken up in dichloromethane and washed with saturated NaHCO3 solution and brine, followed by anhydrous N The residue was extracted with a mixture of petroleum ether and dichloromethane. After filtration, the title compound was obtained as a white solid (100 g, 79% yield). 1 HN MR (500 MHz, CDCl3) δ 7.81 (d, J = 71.1 Hz, 2H), 5.03 (s, 1H), 3.73 (dq, J = 9. 4, 7.1 Hz, 2H), 3.64 (dq, J = 9.4, 7.0 Hz, 2H), 1.25 (t, J = 7.1 Hz, 6H).

[0269] Example 14: Ethyl 2-(diethoxymethyl)thiazole-4-carboxylate (182) Synthesis. [ka]

[0270] 2,2-Diethoxyethanothioamide (100 g, 0.61 m) in ethanol (1 L) ol, 1.0 equiv.) and ethyl bromopyruvate (142 ml, 1.1 mol, 1. To the mixture of 8 equivalents, 90 g of molecular sieves (3 Å) was added. The mixture was refluxed (internal (temperature approximately 60°C), then remove the ethanol on a rotovap and dissolve the residue in dichloromethane. The solid was filtered off, and the filtrate was concentrated and purified by column chromatography (PE / EtOAc 5:1 to 3:1) to obtain the title compound (thiazole carboxylate) as an oil. The compound was obtained as a soluble solid (130 g, 82% yield).

[0271] Example 15: Synthesis of ethyl 2-formylthiazole-4-carboxylate (183) [ka]

[0272] 2-(diethoxymethyl)thiazole-4-carboxylic acid (130g, 0.50mol) To a solution of acetone (1.3 L), 2N HCl (85 mL, 0.165 mol, 0.33 The reaction mixture was refluxed (internal temperature about 60° C.) and TLC analysis showed that the starting material The reaction mixture was monitored until the material was completely consumed (approximately 1-2 hours). The acetone was removed under reduced pressure. The residue was taken up in dichloromethane (1.3 L) and washed with saturated NaHCO3 solution, water, and The solution was washed with brine and dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by recrystallization from petroleum ether and diethyl ether to give the title compound in white. This was obtained as a yellow solid (40 g, 43% yield). 1 H NMR (500 MHz, CDCl3) δ 10.08 - 10.06 (m, 1H), 8.53 - 8.50 (m, 1H), 4.49 (q, J = 7.1 Hz, 2H), 1.44 (t, J = 7.1 Hz, 3H ). MS ESI m / z calculated value C7H8NO3S [M+H] + Measurement 186.01;

[0273] Example 16: 2-((R,E)3-(((S)-tert-butylsulfinyl)imino )-1-hydroxy-4-methylpentyl)thiazole-4-carboxylate ethyl (187 ) synthesis [ka]

[0274] Diisopropylamine (121 mL, 0.86 mol, 4.0 equiv) was added under N2 at -78 °C. A solution of 1000 ml of n-butyllithium (2.5 M, 302 ml) in dry THF was added to The reaction mixture was warmed to 0°C over 30 minutes, after which - Cool to 78°C. (S,E)-2-methyl-N-(3-methylbutan-2-ylidene ) Propane-2-sulfonamide (57 g, 0.3 mol, 1.4 eq) in THF ( 200 mL) was added. Stirring was continued for 1 hour. i Pr)3 (168.5g, 0.6 A solution of 45 mol (3.0 equivalents) in THF (350 mL) was added dropwise. After stirring for 1 hour, 2- Ethyl formylthiazole-4-carboxylate (40 g, 0.215 mol, 1.0 equiv.) A solution of 175 mL of HCl in THF was added dropwise, and the resulting reaction mixture was stirred for 2 hours. The reaction was shown to be complete by precipitation. The reaction was quenched with 500 mL of EtOAc (4×500 mL) and then poured into ice water and The organic phase was washed with water and brine, dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by column chromatography (DCM / EtOAc / PE 2:1:2). Purification gave the title compound as a colorless oil (60 g, 74% yield). 1 H NMR (500 MHz, CDCl3) δ 8.13 (s, 1H), 6.63 (d, J = 8.2 Hz, 1H), 5.20 - 5.11 (m, 1H), 4.43 (q, J = 7.0 Hz, 2H), 3.42 - 3.28 (m, 2H), 2.89 (dt, J = 13.1, 6.5 Hz, 1H), 1.42 (t, J = 7.1 Hz, 3H), 1.33 (s, 9H), 1.25 - 1.22 (m, 6H). MS ESI m / z calculated value C 16 H 26 NaNO 4S2[M+Na] + 397.13, measured value 397.11.

[0275] Example 17: 2-((1R,3R)-3-((S)-1,1-dimethylethylsulfine Ethyl 1-hydroxy-4-methylpentylthiazole-4-carboxylate (1 Synthesis of 88) [ka]

[0276] 2-((R,E)3-(((S)-tert-butylsulfinyl)imino)-1-hydroxybenzoate Ethyl hydroxy-4-methylpentylthiazole-4-carboxylate (23.5g, 62 A THF solution (200 mL) of Ti(OEt)4 (0.7 mmol) was cooled to -45 °C and After the addition was completed, the mixture was stirred for 1 hour. Then NaBH4 (4.75 g, 126 mmol, 2.0 equiv.) was added in one portion. The mixture was stirred at -45°C for 3 hours. TLC analysis showed that some starting material remained. HOAc / THF (v / v 1:4, 25 mL) followed by EtOH (25 mL) The reaction mixture was quenched by pouring it onto ice (100 g) and warming it to room temperature. The reaction mixture was filtered through Celite. Afterwards, the organic phase was separated, washed with water and brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by column chromatography (EtOAc / PE 1:1) to give the title compound. The compound was obtained as a white solid (16.7 g, 71% yield). 1 H NMR (500 MHz, CDCl3) δ 8.10 (s, 1H), 5.51 (d, J = 5.8 Hz, 1H), 5.23 - 5.15 (m, 1H), 4.41 (q, J = 7.0 Hz, 2H), 3.48 - 3.40 (m, 1H), 3.37 (d, J = 8.3 Hz, 1H), 2.29 (t, J = 13.0 Hz, 1 H), 1.95 - 1.87 (m, 1H), 1.73 - 1.67 (m, 1H), 1.40 (t, J = 7.1 Hz, 3H), 1.29 (s, 9H), 0.93 (d, J = 7.3 Hz, 3H), 0.90 (d, J = 7.2 Hz, 3H). MS ESI m / z calculated value C 16 H 28 NaN2O4S2[M+Na] + 399.15, measured value 399.14.

[0277] Example 18: 2-((1R,3R)-3-amino-1-hydroxy-4-methylpentyl ) Synthesis of ethyl thiazole-4-carboxylate (189) [ka]

[0278] At 0°C, 2-((1R,3R)-3-((S)-1,1-dimethylethylsulfinyl) Ethyl (6-hydroxy-1-hydroxy-4-methylpentyl)-thiazole-4-carboxylate 4.00g, 16.0mmol, 1.0 equiv) in ethanol (40mL) A solution of NHCl in dioxane (40 mL) was slowly added. The reaction mixture was allowed to warm to room temperature. The mixture was stirred for 5 hours, then concentrated and treated with petroleum ether dropwise to give the title compound (4. 54 g, 92% yield, was collected and used in the next step.

[0279] Example 19: 2-((1R,3R)-3-((2S,3S)-2-azido-3-methylpentene) (Pentanamido)-1-hydroxy-4-methylpentyl)thiazole-4-carboxylic acid ester Synthesis of Chil(190) [ka]

[0280] (2S,3S)-2-azido-3-methylpentanoic acid (5.03 g, 28.8 mmol) , 2.0 equiv.) in THF (120 mL), cooled to 0° C., and NMM (6.2 mL, 56.0 mmol, 4.0 equiv), isobutyl chloroformate (3.7 mL, 28.8 The reaction mixture was stirred at 0°C for 30 minutes and at room temperature for 1 hour. After that, the temperature was returned to 0°C. 2-((1R,3R)-3-amino-1-hydroxy-4-methyl pentyl)thiazole-4-carboxylate (4.54 g, 14.7 mmol, 1.0 After stirring at 0°C for 30 minutes, the reaction was warmed to room temperature and stirred for 2 hours. The reaction was quenched by adding water at 0°C and extracted three times with ethyl acetate. Wash with 1N HCl, saturated NaHCO3, and brine, dry over anhydrous Na2SO4, The residue was purified by column chromatography (0-30% EtOAc / PE). Purification gave the title compound (4.55 g, 74% yield) as a white solid.

[0281] Example 20: 2-((1R,3R)-3-((2S,3S)-2-azido-3-methylpentene) 4-methyl-1-((triethylsilyl)oxy)pentyl)thiazolinone Synthesis of ethyl benzoyl-4-carboxylate (191) [ka]

[0282] At 0°C, 2-((1R,3R)-3-((2S,3S)-2-azido-3-methylpentene Tanamido)-1-hydroxy-4-methylpentyl)thiazole-4-carboxylic acid ethyl ester To a solution of 5.30 g (12.8 mmol, 1.0 equiv.) in CHCl (50 mL), Imidazole (1.75 g, 25.6 mmol, 2.0 equiv.) followed by chlorotriethyl Silane (4.3 mL, 25.6 mmol, 2.0 equiv) was added. The reaction mixture was stirred for 1 h. The mixture was allowed to warm to room temperature and stirred for an additional hour. Brine was added to the reaction mixture and the organic phase was separated. The aqueous phase was extracted with EtOAc. The combined organic phases were dried, filtered, concentrated under reduced pressure, and Column chromatography using a gradient of 15-35% EtOAc in petroleum ether and purified to give the title compound as a white solid (6.70 g, 99% yield). 1 H NMR (50 0 MHz, CDCl3) δ 8.12 (s, 1H), 6.75 (d, J = 8.0 Hz, 1H), 5.20 - 5.12 (m, 1H), 4. 44 (q, J = 7.0 Hz, 2H), 4.06 - 3.97 (m, 1H), 3.87 (d, J = 3.8 Hz, 1H), 2.14 (d, J = 3.8 Hz, 1H), 2.01 - 1.91 (m, 3H), 1.42 (t, J = 7.1 Hz, 3H), 1.34 - 1.25 (m, 2H), 1.06 (d, J = 6.8 Hz, 3H), 1.00 - 0.93 (m, 18H), 0.88 (dd, J = 19.1, 6.8 Hz, 6H). MS ESI m / z calculation value C 24 H 44 N5O4SSi [M+H] + 526.28, Measurement 526.28.

[0283] Example 21: 2-((1R,3R)-3-((2S,3S)-2-azido-N,3-dimethyl 4-methyl-1-((triethylsilyl)oxy)pentyl)thio Synthesis of ethyl azole-4-carboxylate (192) [ka]

[0284] 2-((1R,3R)-3-((2S,3S)-2-azido-3-methylpentanamine 4-methyl-1-((triethylsilyl)oxy)pentyl)thiazole-4-carboxylate Ethyl carboxylate (5.20 g, 9.9 mmol, 1.0 equiv.) in THF (50 mL) was cooled to -45 °C and KHMDS (1 M in toluene, 23.8 mL, 23.8 mmol, The resulting mixture was stirred at -45°C for 20 minutes. Then, methyl iodide was added. ethanol (1.85 mL, 29.7 mmol, 3.0 equiv.) was added and the reaction mixture was stirred for 4.5 h. The reaction was allowed to warm to room temperature, at which point the reaction was quenched with EtOH (10 mL). The aqueous phase was diluted with tOAc (250 mL) and washed with brine (100 mL). The organic phase was dried, filtered, concentrated, and diluted with 15 ml of petroleum ether. Purification by column chromatography with a gradient of 35% EtOAc afforded the title compound. Obtained as a pale yellow oil (3.33 g, 63% yield). 1 H NMR (500 MHz, CDCl3) δ 8 .09 (s, 1H), 4.95 (d, J = 6.6 Hz, 1H),4.41 (q, J = 7.1 Hz, 2H), 3.56 (d, J = 9.5 Hz, 1H), 2.98 (s, 3H), 2.27 - 2.06 (m, 4H), 1.83 - 1.70 (m, 2H), 1.41 (t, J = 7 .2 Hz, 3H), 1.29 (ddd, J = 8.9, 6.8, 1.6 Hz, 3H), 1.01 (d, J = 6.6 Hz, 3H), 0.96 (dt, J = 8.0, 2.9 Hz, 15H), 0.92 (d, J = 6.6 Hz, 3H), 0.90 (d, J = 6.7 Hz,3H). MS ESI m / z calculation value C 25 H 46 N5O4SSi [M+H] + 540.30, Measurement 540.30.

[0285] Example 22: 2-((3S,6R,8R)-3-((S)-sec-butyl)-10,1 0-Diethyl-6-isopropyl-5-methyl-1-((R)-1-methylpiperidine- 2-yl)-1,4-dioxo-9-oxa-2,5-diazo-10-siladodecane-8 Synthesis of ethyl thiazole-4-carboxylate [ka]

[0286] Dry Pd / C (10wt%, 300mg) and 2-(1R,3R)-3-((2S,3 S)-2-Azido-N,3-dimethylpentanamido-4-methyl-1-(triethylsilyl) Ethyl (( ... ol) in EtOAc The reaction mixture was stirred under a hydrogen atmosphere for 27 hours and then added to the carboxylate. The mixture was filtered through a plug of silica gel and the filter pad was washed with EtOAc. Concentrate the fraction and eluate by column chromatography with a gradient of 0-5% methanol in EtOAc. The title compound was obtained (3.90 g, 86% yield). Value C 32 H 59 N4O5SSi [M+H]+ 639.39, measured value 639.39.

[0287] Example 23: 2-((1R,3R)-3-((2S,3S)-N,3-dimethyl-2-( (R)-1-Methylpiperidine-2-carboxamido)pentanamido)-1-hydroxy Synthesis of ethyl thiazole-4-carboxylate [ka]

[0288] 2-((3S,6R,8R)-3-((S)-sec-butyl)-10,10-diethyl 6-isopropyl-5-methyl-1-((R)-1-methylpiperidin-2-yl) -1,4-dioxo-9-oxa-2,5-diaza-10-siladodecan-8-yl)thio Ethyl azole-4-carboxylate (3.90 g, 6.1 mmol) was dissolved in deoxygenated AcOH / water / THF (v / v / v 3:1:1, 100 mL) and stirred at room temperature for 48 hours. The reaction mixture was then concentrated and purified by column chromatography (2:98 to 15:85 MeOH). / EtOAc) to give the title compound (2.50 g, 72% yield over two steps) . MS ESI m / z calculation value C 26 H 45 N4O5S [M+H] + 525.30, measured value 525.33.

[0289] Example 24: 2-((1R,3R)-3-((2S,3S)-N,3-dimethyl-2-( (R)-1-Methylpiperidine-2-carboxamido)pentanamido)-1-hydroxy Synthesis of (4-methylpentyl)thiazole-4-carboxylic acid [ka]

[0290] At 0 °C, aqueous LiOH (0.4 N, 47.7 mL, 19.1 mmol, 4.0 equiv.) 2-((1R,3R)-3-((2S,3S)-N,3-dimethyl-2-((R)- 1-Methylpiperidine-2-carboxamide)pentanamide)-1-hydroxy-4- methylpentyl) ethyl thiazole-4-carboxylate (2.50 g, 4.76 mmol, The reaction mixture was stirred at room temperature for 2 hours. The mixture was stirred and then concentrated. Column chromatography (100% CH2Cl2, then CH 2Cl2 / MeOH / NH4OH 80:20:1) to give the title compound (2.36 g, The compound was obtained in a 99% yield as an amorphous solid. MS ESI m / z calculated C 24 H 41 N4O5S [M+H] + 497.27, measurement Fixed price 497.28.

[0291] Example 25: 2-((1R,3R)-1-acetoxy-3-((2S,3S)-N,3- Dimethyl-2-((R)-1-methylpiperidine-2-carboxamido)pentanamide Synthesis of 4-methylpentylthiazole-4-carboxylic acid [ka]

[0292] At 0°C, 2-((1R,3R)-3-((2S,3S)-N,3-dimethyl-2-(( R)-1-Methylpiperidine-2-carboxamido)pentanamido)-1-hydroxy -4-methylpentyl)thiazole-4-carboxylic acid (2.36 g, 4.75 mmol) To the pyridine solution (50 mL) of acetic anhydride (2.25 mL, 24 mmol) was slowly added. The reaction mixture was allowed to warm to room temperature over 2 hours and stirred at room temperature for 24 hours. The reaction was concentrated. The residue was analyzed by reverse phase HPLC (C 18 column, 10-90% acetonitrile / water) and The title compound was obtained as an amorphous white solid (2.25 g, 88% yield). MS ESI m / z calculated C 26 H 43 N4O6S [M+H] +539.28, Measurement 539.28.

[0293] Example 26: (1R,3R)-3-((2S,3S)-N,3-dimethyl-2-((R) -1-methylpiperidine-2-carboxamido)pentanamido)-4-methyl-1-( Synthesis of 4-(perfluorobenzoyl)thiazol-2-yl)pentylacetic acid (294) . [ka]

[0294] At 0°C, 2-((1R,3R)-1-acetoxy-3-((2S,3S)-N,3-dimethyl ethyl-2-((R)-1-methylpiperidine-2-carboxamide)pentanamide)- 4-methylpentyl)thiazole-4-carboxylic acid (86 mg, 0.16 mmol, 1. 0 equiv.) in dichloromethane, ol, 1.5 eq) and N,N'-diisopropylcarbodiimide (22 mg, 0.17 5 mmol, 1.1 equiv.) was added. The reaction mixture was allowed to warm to room temperature and stirred overnight. The solvent was After removal under reduced pressure, the reaction mixture was diluted with EtOAc (2 mL) and then filtered through Celite. The filtrate was concentrated to give the title compound, which was used directly without further purification.

[0295] Example 27: 3-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)propanol Synthesis of tert-butyl phosphate [ka]

[0296] 2,2'-(ethane-1,2-diylbis(oxy)) in anhydrous THF (200 mL) A solution of sodium in diethanol (55.0 mL, 410.75 mmol, 3.0 equiv.) The mixture was stirred until the sodium hydroxide disappeared, and then acrylic acid tert-butyl ether (0.1 g) was added. t-Butyl ether (20.0 mL, 137.79 mmol, 1.0 equiv.) was added dropwise. The mixture was stirred overnight and then quenched with HCl solution (20.0 mL, 1N) at 0° C. Remove by rotary evaporation, add brine (300 mL), and the resulting mixture is diluted with Et The organic layer was washed with brine (3 x 100 mL) and extracted with HCl (3 x 100 mL). Dry over anhydrous NaSO, filter, and concentrate to a colorless oil (30.20 g, yield 79.0%), which was used without further purification. MS ESI m / z calculated C 13 H 27 O6[ M + H] + 278.1729, measurement 278.1730.

[0297] Example 28: 3-(2-(2-(2-tosyloxy)ethoxy)ethoxy)propanoic acid Synthesis of ert-butyl [ka]

[0298] At 0°C, 3-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)propane tert-Butyl acetate (30.20 g, 108.5 mmol, 1.0 equiv.) and TsCl( 41.37 g, 217.0 mmol, 2.0 equiv) in anhydrous DCM (220 mL), TEA (30.0 mL, 217.0 mmol, 2.0 equiv.) was added and the mixture was stirred at room temperature. Stir overnight, then wash with water (3 x 300 mL) and brine (300 mL) and add anhydrous NaCl. Drying with 2SO4, filtering, concentrating, and eluting with SiO2 column chromatography (3:1 hexahydrate) Purification by hexane / EtOAc gave a colorless oil (39.4 g, 84.0% yield). MS ESI m / z calculation value C 20 H 33 O8S [M+H] + 433.1818, measured value 433.2838.

[0299] Example 29: 3-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)propanoic acid Synthesis of tert-butyl. [ka]

[0300] 3-(2-(2-(2-tosyloxy)ethoxy)ethoxy)ethoxy)-3-(2-(2-tosyloxy)eth ... A solution of tert-butyl hydroxypropanoate (39.4 g, 91.1 mmol, 1.0 equiv.) To the solution was added NaN3 (20.67 g, 316.6 mmol, 3.5 equivalents). Stirred at room temperature overnight. Add water (500 mL) and extract with EtOAc (3 x 300 mL). The combined organic layers were washed with water (3 x 900 mL) and brine (900 mL) and Drying with NaSO was performed, and the mixture was then purified by SiO column chromatography (5:1 hexane / EtO Ac), filtered, concentrated and purified to give a pale yellow oil (23.8 g, 85.53% yield).

[0301] Example 30: 3-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)propanoic acid Synthesis of tert-butyl. [ka]

[0302] Raney Ni (7.5 g, suspended in water) was dissolved in water (3 times) and isopropyl alcohol (3 times). The compound (147) (5.0 g, 16.5 mmol) in isopropyl alcohol was washed with The mixture was stirred under a H2 balloon at room temperature for 16 hours, then placed on a Celite pad. The filtrate was concentrated and the column was then washed with isopropyl alcohol. Purification by chromatography (5-25% MeOH / DCM) gave a pale yellow oil (2.60 g, The yield was 57%. MS ESI m / z calculated value C 13 H 28 NO5[M+H] + Measured 279.19;

[0303] Example 31: Synthesis of 2-(2-(dibenzylamino)ethoxy)ethanol (298) [ka]

[0304] 2-(2-aminoethoxy)ethanol (21.0 mL) in acetonitrile (350 mL) 0 g, 200 mmol, 1.0 equiv.) and K2CO3 (83.00 g, 600 mmol, To the resulting solution (3.0 equiv.), BnBr (57.0 mL, 480 mmol, 2.4 equiv.) was added. The mixture was refluxed overnight, water (1 L) was added and extracted with EtOAc (3 x 300 mL). The combined organic layers were washed with brine (1000 mL), dried over anhydrous Na2SO4, and Filtration, concentration, and purification by O2 column chromatography (4:1 hexane / EtOAc) The resulting product was a colorless oil (50.97 g, 89.2% yield). MS ESI m / z calculated C 18 H 23 NO2Na [M+Na] + 309.1729, measured value 309.1967.

[0305] Example 32: 3-(2-(2-(dibenzylamino)ethoxy)ethoxy)propanoic acid Synthesis of ert-butyl (300). [ka]

[0306] 2-(2-(dibenzylamino)ethoxy)ethanol (4 7.17 g, 165.3 mmol, 1.0 equiv.), tert-butyl acrylate (72. 0 mL, 495.9 mmol, 3.0 equiv.), and n-Bu4NI (6.10 g, 16. 53 mmol, 0.1 equiv.) was added to a mixture of sodium hydroxide solution (300 mL, 50%) was added. The mixture was stirred overnight. The organic layer was separated and the aqueous layer was diluted with EtOAc (3×100 mL The organic layer was washed with water (3×100 mL) and brine (300 mL) and The extract was dried over water, Na2SO4, filtered, concentrated, and purified by SiO2 column chromatography ( Purification by hexanes / EtOAc (7:1) gave a colorless oil (61.08 g, yield rate 89.4%). MS ESI m / z calculation value C 25 H 36 NO4[M+H] + 414.2566, measured value 414.2384.

[0307] Example 33: tert-Butyl 3-(2-(2-aminoethoxy)ethoxy)propanoate Synthesis of (301) [ka]

[0308] In a hydrogenation bottle, 3-(2-(2-(dibenzylamino)ethoxy)ethoxy)pro tert-Butyl pantothenate (20.00 g, 48.36 mmol, 1.0 equiv.) in THF( Pd / C (2.00 g, 10 wt%) was added to a mixture of Pd / C (3.00 g, 10 wt%) and MeOH (60 mL). , 50% wet) was added. The mixture was shaken overnight and filtered through Celite (filter aid). The filtrate was concentrated to give a colorless oil (10.58 g, yield 93.8%). MS ESI m / z calculation Value C 11 H 24 NO4[M+H] + 234.1627, measured value 234.1810.

[0309] Example 34: tert-Butyl 3-(2-(2-hydroxyethoxy)ethoxy)propanoate Synthesis of Chil [ka]

[0310] of 2,2'-oxydiethanol (19.7 mL, 206.7 mmol, 3.0 equiv.) Sodium (0.1 g) was added to anhydrous THF solution (100 mL). The mixture was stirred with tert-butyl acrylate (10.0 mL, 68.9 mmol) The mixture was stirred overnight, brine (200 mL) was added, and E Extract with tOAc (3 x 100 mL). Wash the organic layer with brine (3 x 300 mL). The extract was dried over anhydrous Na2SO4 and purified by column chromatography (1:1 hexane / Filtration, concentration, and purification with EtOAc gave a colorless oil (8.10 g, yield 49. 4%). MS ESI m / z calculation value C 11 H 23 O5 [M+H] + 235.1467, measured value 235.1667.

[0311] Example 35: 3-(2-(2-(tosyloxy)ethoxy)ethoxy)propanoic acid tert-butyl ester Synthesis of t-butyl [ka]

[0312] At 0°C, tert-butyl 3-(2-(2-hydroxyethoxy)ethoxy)propanoate (6.24 g, 26.63 mmol, 1.0 equiv.) and TsCl (10.15 g, 53 A solution of 0.27 mmol, 2.0 equiv) in anhydrous DCM (50 mL) was added to pyridine (4.3 mL , 53.27 mmol, 2.0 equiv) was added. The mixture was stirred at room temperature overnight and then water ( The aqueous layer was washed with DCM (3 x 50 mL) and the combined organic layer was The mixture was washed with water (300 mL), dried over anhydrous Na2SO4, and then subjected to SiO2 column chromatography. Filtration, concentration, and purification by elution with hexane / EtOAc (5:1) gave a colorless oil. Obtained (6.33 g, 61.3% yield). MS ESI m / z calculated C 18 H 27 O7S [M+H] + 389.1556, measurement Fixed price 389.2809.

[0313] Example 36: tert-Butyl 3-(2-(2-azidoethoxy)ethoxy)propanoate Synthesis of [ka]

[0314] tert-Butyl 3-(2-(2-(tosyloxy)ethoxy)ethoxy)propanoate (5.80 g, 14.93 mmol, 1.0 equiv.) in anhydrous DMF (20 mL) aN3 (5.02 g, 77.22 mmol, 5.0 equiv.) was added, and the mixture was allowed to stand at room temperature overnight. Water (120 mL) was added and extracted with EtOAc (3 x 50 mL). The organic layer was washed with water (3 x 150 mL) and brine (150 mL) and diluted with anhydrous NaSO. The resulting mixture was dried over 4°C and subjected to SiO column chromatography (5:1 hexane / EtOAc). The mixture was filtered, concentrated, and purified to give a colorless oil (3.73 g, 69.6% yield). MS ESI Calculated m / z value C 11 H 22 O3N4Na[M+H] + 260.1532, measurement 260.2259.

[0315] Example 37: tert-butyl 3-(2-(2-aminoethoxy)ethoxy)propanoate Synthesis of [ka]

[0316] tert-Butyl 3-(2-(2-azidoethoxy)ethoxy)propanoate (0.18 g, 0.69 mmol) was dissolved in MeOH (3.0 mL, containing 60 μL of HCl). The catalyst was then hydrogenated in the presence of Pd / C (10 wt%, 20 mg) under a H2 balloon. The medium was filtered through a pad of Celite and the pad was washed with MeOH. The filtrate was concentrated to a colorless An oil was obtained (0.15 g, 93% yield). MS ESI m / z calculated C 11 H 24 NO4[M+H] + 234.16; Measurement value: 234.14.

[0317] Example 38: Synthesis of 3-(2-(2-azidoethoxy)ethoxy)propanoic acid [ka]

[0318] tert-Butyl 3-(2-(2-azidoethoxy)ethoxy)propanoate (2.51 g, 9.68 mmol) was dissolved in 1,4-dioxane and treated with 10 mL of concentrated HCl at room temperature. The mixture was stirred for 35 min and diluted with EtOH (30 mL) and toluene (30 mL). The mixture was diluted and concentrated under vacuum. A solution of methanol (5%-10%) and 1% formic acid in methylene chloride was added. The crude mixture was purified on silica gel using as eluent to give the title compound (1.63 g , yield 83%). ESI MS m / z C7H 12 N3O4[MH] - , calculated value 202.06, measured value 202.30.

[0319] Example 39: 2,5-Dioxopyrrolidin-1-yl 3-(2-(2-azidoethoxy) Synthesis of )ethoxy)propanoate [ka]

[0320] 3-(2-(2-azidoethoxy)ethoxy)propanoic acid (1.60 g, 7.87 mm ol) in dichloromethane (30 mL), NHS (1.08 g, 9.39 mmol) , EDC (3.60 g, 18.75 mmol) was added. After 8 hours, TLC analysis was performed. The reaction was shown to be complete, and a solution of ethyl acetate (5%-10%) in methylene chloride was added. The reaction mixture was concentrated and purified on silica gel using as the eluent to give the title compound ( 1.93g, 82% yield) ESI MS m / z C 11 H 17 N4O6[M+H]+, calculated 301.11, found 301.20.

[0321] Example 40: (S)-15-azido-5-isopropyl-4,7-dioxo-10,13 Synthesis of -dioxa-3,6-diazapentadacan-1-oic acid [ka]

[0322] A mixture of DMA (50 mL) and 0.1 M NaH2PO4 (50 mL, pH 7.5) (S)-2-(2-amino-3-methylbutanamido)acetic acid (Val-Gly) (1 0.01g, 5.80mmol) to 2,5-dioxopyrrolidin-1-yl 3-(2-( 2-Azidoethoxy)ethoxy)propanoic acid (1.90 g, 6.33 g) was added. The mixture was stirred for 4 hours, evaporated under vacuum, and mixed with methanol in methylene chloride containing 0.5% acetic acid. The mixture (5%-15%) was purified on silica gel as eluent to give the title compound (1.52 g, yield 73%). ESI MS m / z C 14 H 26 N5O6[M+H] + , calculated value 360.18, measured value 360.40.

[0323] Example 41: (S)-2,5-dioxopyrrolidine-1-yl 15-azido-5-isopropyl propyl-4,7-dioxo-10,13-dioxa-3,6-diazapentadacane-1- Synthesis of oate [ka]

[0324] (S)-15-Azido-5-isopropyl-4,7-dioxo-10,13-dioxa -3,6-diazapentadecan-1-oic acid (1.50 g, 4.17 mmol) dichloromethane To a solution (40 mL) of ethanol, NHS (0.88 g, 7.61 mmol) and EDC (2.6 0 g, 17.90 mmol) was added with stirring. After 8 hours, TLC analysis showed The reaction was shown to be complete, and ethyl acetate (5% to 2%) in methylene chloride was used as the eluent. The reaction mixture was concentrated onto silica gel and purified using a mixture of 0% HCl to give the title compound ( 1.48g, 78% yield) ESI MS m / z C 18 H 29 N6O8[M+H] + , Calculated value 457.20, Measured value 457.50 .

[0325] Example 42: Synthesis of 4-(((benzyloxy)carbonyl)amino)butanoic acid [ka]

[0326] 4-aminobutyric acid (7.5 g, 75 mmol) and NaOH (6 g, 150 mmol) A solution of CbzCl (16.1 g, 95 mmol) in HO (40 mL) was cooled to 0 °C and The reaction was treated dropwise with HF solution (32 mL). After 1 h, the reaction was allowed to warm to room temperature and stirred for 3 h. The THF was removed under vacuum and the pH was adjusted to 1.5 by adding 6N HCl. Extraction with ethyl acetate followed by washing the organic layer with brine, drying and concentration gave the title compound. (16.4 g, 92% yield). MS ESI m / z calculated C 12 H 16 NO5[M+H] + 238.10, measured value 238. 08.

[0327] Example 43: 4-(((benzoyloxy)carbonyl)amino)butanoic acid tert-butyl Synthesis of Chil [ka]

[0328] 4-((benzyloxy)carbonyl)amino)butanoic acid (16.4 g, 69.2 mm ol) and t-BuOH (15.4 g, 208 mmol) in DCM (100 mL) , DMAP (0.8 g, 6.56 mmol) and DCC (17.1 g, 83 mmol) After stirring overnight at room temperature, the reaction mixture was filtered and concentrated. The residue was dissolved in ethyl acetate. , 1N HCl, brine, and dried over Na2SO4. Column chromatograph Concentration and purification by elution with 10-50% EtOAc / hexanes gave the title compound. (7.5 g, 37% yield). MS ESI m / z calculated C 16 H 23 NO4Na [M+Na] + 316.16, measurement 316 .13.

[0329] Example 44: Synthesis of 4-aminobutanoic acid tert-butyl ester [ka]

[0330] 4-((benzyloxy)carbonyl)amino acid tert-butyl ester (560 mg, 1. 91 mmol) was dissolved in MeOH (50 mL) and Pd / C catalyst (10 wt%, 100 m g) and hydrogenated (1 atm) at room temperature for 3 hours. The catalyst was filtered off and all volatiles were removed. The was removed in vacuo to give the title compound (272 mg, 90% yield). MS ESI m / z calculated C8H 18 NO2[M+H] + 160.13, Measurement 160.13.

[0331] Example 45: tert-Butyl 2-(triphenylphosphoranylidene)propanoate (20 Synthesis of 6) [ka]

[0332] tert-Butyl 2-bromopropanoate (15. 5g, 74.1mmol, 1.0 eq) and triphenylphosphine (19.4g, 74 A mixture of 1.0 eq. (0.1 mmol, 1.0 eq.) was stirred at room temperature for 18 hours, and then acetonitrile was added under reduced pressure. The toluene was removed and toluene was added to decompose the white precipitate. The solid was dissolved in dichloromethane (100 mL) and transferred to a separatory funnel. 00 mL) was added to the funnel and the organic layer immediately turned yellow. The organic layer was separated and the aqueous layer was washed once with water. Extraction was performed with dichloromethane (30 mL). The dichloromethane layers were combined and washed with brine (50 mL). L), then dried over Na2SO4, filtered and concentrated to give the ylide as a yellow solid ( The weight was 16.8g, 58%).

[0333] Example 46: (S)-methyl 3-(4-(benzoyloxy)phenyl)-2-((t Synthesis of (ert-butoxycarbonyl)amino)propanoic acid (203) [ka]

[0334] Boc-L-Tyr-OMe (20.0 g, 67.7 mmHg) in acetone (100 mL) ol, 1.0 eq.), K2CO3 (14.0 g, 101.6 mmol, 1.5 eq.), and A mixture of BnBr (10.5g, 6.77mmol, 0.1eq) and KI (1.12g, 6.77mmol, 0.1eq) was added. mL, 81.3 mmol, 1.2 equiv.) was slowly added. The mixture was then refluxed overnight. Water (250 mL) was added and the reaction mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (300 mL), dried over anhydrous Na2SO4, and filtered. The extract was concentrated and purified by SiO2 column chromatography (4:1 hexane / EtOAc). The title compound was obtained as a white solid (26.12 g, 99% yield). 1 H NMR (50 0 MHz, CDCl3) δ 7.44 - 7.41 (m, 2H), 7.41 - 7.36 (m, 2H), 7.35 - 7.30 (m, 1H), 7.04 (d, J = 8.5 Hz, 2H), 6.93 - 6.89 (m, 2H), 5.04 (s, 2H), 4.97 (d, J = 7.7 Hz , 1H), 4.55 (d, J = 6.9 Hz, 1H), 3.71 (s, 3H), 3.03 (dd, J = 14.4, 5.7 Hz, 2H), 1.44 (d, J = 18.6 Hz, 10H). MS ESI m / z calculated value C 22 H 27 NO5Na [M+Na] + 408.18, measurement 408 .11.

[0335] Example 47: (S)-tert-butyl (1-(4-benzyloxy)phenyl)- Synthesis of 3-oxopropan-2-yl)carbamate (204) [ka]

[0336] At -78°C, (S)-methyl 3-(4-(benzyloxy)phenyl)-2-((t (ert-butoxycarbonyl)amino)propanoate (26.1g, 67.8mmol , 1.0 equiv.) in anhydrous dichloromethane (450 mL) with DIBAL (1 in hexane) A solution of 1.0M HCl (163 mL, 2.2 equiv.) was added over 1 hour. The mixture was stirred at -78°C for 3 hours. Then quench with 50 mL of ethanol. Add 1N HCl dropwise until a pH of 4 is reached. The resulting mixture was warmed to 0° C. The layers were separated and the aqueous layer was further diluted with EtOAc (3×100 The combined organic solution was washed with brine, dried over anhydrous Na2SO4, and concentrated Trituration with PE / EtOAc and filtration gave the title compound as a white solid (18.3%). g, yield 76%). MS ESI m / z calculation value C 22 H 27 NO5Na [M+Na] + 378.11, Measurement 378.11.

[0337] Example 48: (S,Z)-tert-butyl 5-(4-(benzyloxy)phenyl) -4-((tert-butoxycarbonyl)amino)-2-methylpent-2-enoate Synthesis of 207 [ka]

[0338] (S)-tert-butyl (1-(4-benzyloxy)phenyl)-3-oxo propan-2-yl)carbamate (0.84 g, 2 mmol, 1.0 equiv.) was added to dry Dissolved in chloromethane (50 mL). 2-(triphenylphosphoranylidene)propanoic acid tert-Butyl ether (1.6 g, 4 mmol, 2.0 equiv.) was added and the solution was stirred at room temperature for 1.5 min. The mixture was stirred for 10 hours and judged complete by TLC. 0% EtOAc / hexanes) to give the title compound (1.16 g, 98% yield) .

[0339] Example 49: (4R)-tert-butyl 4-((tert-butoxycarbonyl)a Synthesis of 5-(4-hydroxyphenyl)-2-methylpentanoate [ka]

[0340] (S,Z)-tert-butyl 5-(4-(benzyloxy)phenyl)-4-(( tert-Butoxycarbonyl)amino)-2-methylpent-2-enoate (467 mg, 1 mmol) was dissolved in methanol (30 mL) and Pd / C catalyst (10 wt%, 2 The catalyst was removed by filtration, and the filtrate was then heated under reduced pressure. The mixture was concentrated to give the title compound (379 mg, 99% yield).

[0341] Example 50: (4R)-tert-butyl 4-((tert-butoxycarbonyl)a (amino)-5-(4-hydroxy-3-nitrophenyl)-2-methylpentanoate Growth [ka]

[0342] (4R)-tert-butyl 4-((tert-butoxycarbonyl)amino)-5 -(4-hydroxyphenyl)-2-methylpentanoate (379 mg, 1 mmol, 1.0 equiv.) was dissolved in THF (20 mL). A solution of 5 mg, 3 mmol, 3.0 equiv) in THF (2 mL) was added. The reaction was stirred at room temperature for 3 The mixture was stirred for 1 hour, poured into water, extracted with EtOAc (2 x 50 mL), and the combined organic phases were The mixture was washed with ethanol (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The title compound was purified by column chromatography (10-50% EtOAc / hexanes). Obtained (300 mg, 71% yield).

[0343] Example 51: (4R)-tert-butyl 5-(3-amino-4-hydroxyphenyl) )-4-((tert-butoxycarbonyl)amino)-2-methylpentanoate(2 Synthesis of 10) [ka]

[0344] (4R)-tert-butyl 4-((tert-butoxycarbonyl)amino)-5 -(4-hydroxy-3-nitrophenyl)-2-methylpentanoate (200 mg, 0.47 mmol) in EtOAc (30 mL) and palladium catalyst (10% on carbon) The mixture was mixed with 100 mg of methylcellulose and then hydrogenated (1 atm) at room temperature for 2 hours. All volatiles were removed in vacuo to give the title compound (185 mg, 99%).

[0345] Alternatively, (4R)-tert-butyl 4-((tert-butoxycarbonyl)a (amino)-5-(4-hydroxy-3-nitrophenyl)-2-methylpentanoate (5 6 mg, 0.132 mmol) was dissolved in EtOAc (20 mL) and Pd / C catalyst (10 The mixture was mixed with 50 mg of methylcellulose (wt%) and then hydrogenated (1 atm) at room temperature for 3 hours. All volatiles were removed under vacuum to give the title compound (52 mg, 99% yield). MS E SI m / z calculated value C 21 H 35 N2O5[M+H] +395.25, measured value 395.26.

[0346] Example 52: (4R)-tert-butyl 4-((tert-butoxycarbonyl)a (amino)-5-(4-((tert-butyldimethylsilyl)oxy)-3-nitrophenyl Synthesis of methyl-2-methylpentanoate. [ka]

[0347] (4R)-tert-butyl 4-((tert-butoxycarbonyl)amino)-5 -(4-hydroxy-3-nitrophenyl)-2-methylpentanoate (424 mg, A solution of 1 mmol of imidazole (408 mg, 6 mmol) and and tert-butylchlorodimethylsilane (602 mg, 4 mmol) were added. The resulting solution was stirred at room temperature for 3 hours, after which the reaction mixture was washed with brine (50 mL) and Drying with anhydrous Na2SO4, concentration, and column chromatography (10% to 30% E tOAc / hexane) to give the title compound (344 mg, 64% yield) .

[0348] Example 53: (4R)-tert-butyl 5-(3-amino-4-((tert-butyl) ((dimethylsilyl)oxy)phenyl)-4-((tert-butoxycarbonyl)amino Synthesis of 2-methylpentanoate (215). [ka]

[0349] (4R)-tert-butyl 4-((tert-butoxycarbonyl)amino)-5 -(4-((tert-butyldimethylsilyl)oxy)-3-nitrophenyl)-2- Methyl pentanoate (200 mg, 0.37 mmol) was dissolved in EtOAc (30 mL). The solution was dissolved, mixed with palladium catalyst (10 wt% on carbon, 100 mg), and heated in hydrogen at room temperature for 2 hours. The catalyst was filtered off and all volatiles were removed under vacuum to give the title compound. (187 mg, 99% yield).

[0350] Example 54: (S)-tert-butyl 2-(hydroxymethyl)pyrrolidine-1-carboxylate Synthesis of carboxylates. [ka]

[0351] Boc-L-proline (10.0 g, 46.4 mmol) dissolved in 50 mL of THF The mixture was cooled to 0°C and a solution of BH3 in THF (1.0 M, 46.4 mL) was carefully added. The mixture was stirred at 0°C for 1.5 hours, then poured onto ice water and extracted with ethyl acetate. The organic layer was washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. Concentration gave the title compound as a white solid (8.50 g, 91% yield). 1 H NMR (500 MHz, CDCl3) δ 3.94 (dd, J = 4.9, 2.7 Hz, 2H), 3.60 (ddd, J = 18.7, 11.9, 9.3 Hz , 2H), 3.49-3.37 (m, 1H), 3.34-3.23 (m, 1H), 2.06-1.91 (m, 1H), 1.89-1.69 (m, 2H) ), 1.65-1.51 (m, 1H), 1.49- .40 (m, 9H).

[0352] Example 55: (S)-tert-butyl 2-formylpyrrolidine-1-carboxylate Synthesis of to. [ka]

[0353] (S)-tert-butyl 2-(hydroxybenzoate) in dimethyl sulfoxide (90 mL) To a solution of methyl)pyrrolidine-1-carboxylate (13.0 g, 64.6 mmol) Triethylamine (40 mL) was added and stirring was continued for 15 minutes. The mixture was cooled in an ice bath. , sulfur trioxide-pyridine complex (35.98 g, 226 mmol) was added in small portions over 40 minutes. The reaction mixture was allowed to warm to room temperature and stirred for 2.5 hours. After adding ice (250 g), The mixture was extracted with dichloromethane (150 mL x 3). The organic phase was diluted with 50% citric acid solution. (150 mL), water (150 mL), saturated sodium bicarbonate solution (150 mL), and Washed with brine (150 mL) and dried over anhydrous Na2SO4. Removed the solvent in vacuo. This gave the title aldehyde as a thick oil (10.4 g, 81% yield), which was further purified by Used without purification. 1 H NMR (500 MHz, CDCl3) δ 9.45 (s, 1H), 4.04 (s, 1H), 3.53 (dd, J = 14.4, 8.0 Hz, 2H), 2.00 - 1.82 (m, 4H), 1.44 (d, J = 22.6 Hz, 9H) .

[0354] Example 56: (4R,5S)-4-methyl-5-phenyl-3-propionyloxazolidinyl Synthesis of din-2-one. [ka]

[0355] Under N2, at -78 °C, 4-methyl-5-phenyloxazolidin-2-one (8.0 g , 45.17 mmol) in THF (100 mL) with n-butyllithium (21.6 A hexane solution of 1 mL of 2.2 M hexane (47.43 mmol) was added dropwise. The solution was heated at -78 °C for 1 This was maintained for 1 hour, and then propionyl chloride (4.4 mL, 50.59 mmol) was added slowly. The reaction mixture was warmed to -50°C and stirred for 2 hours, then added with a saturated solution of ammonium chloride (100 mL). The organic solvent was removed in vacuo and the resulting solution The organic layer was extracted with ethyl acetate (3×100 mL). 0 mL) and brine (100 mL), dried over Na2SO4, filtered, and The mixture was concentrated in vacuo, and the residue was purified by column chromatography (20% ethyl acetate / hexane). Purification gave the title compound as a thick oil (10.5 g, 98% yield). 1 H NMR (50 0 MHz, CDCl3) δ 7.45 - 7.34 (m, 3H), 7.30 (d, J = 7.0 Hz, 2H), 5.67 (d, J = 7.3 Hz, 1H), 4.82 - 4.70 (m, 1H), 2.97 (dd, J = 19.0, 7.4 Hz, 2H), 1.19 (t, J = 7.4 Hz, 3H), 0.90 (d, J = 6.6 Hz, 3H).

[0356] Example 57: (S)-tert-butyl 2-((1R,2R)-1-hydroxy-2- Methyl-3-((4R,5S)-4-methyl-2-oxo-5-phenyloxazolidine Synthesis of (3-yl)-3-oxopropyl)pyrrolidine-1-carboxylate [ka]

[0357] At 0°C, (4R,4S)-4-methyl-5-phenyl-3-propionyloxazolidinium To a solution of benzophenone (9.40 g, 40.4 mmol) in dichloromethane (60 mL), EtN (6.45 mL, 46.64 mmol) was added. Then, 1 in dichloromethane was added. Dibutylboron trifluoromethanesulfonate (42 mL, 42 mmol) was added. The mixture was stirred at 0° C. for 45 min, cooled to −70° C., and then diluted with dichloromethane (40 mL). (S)-tert-butyl 2-formylpyrrolidine-1-carboxylate (4. The reaction mixture was stirred at -70°C for 2 hours. The mixture was stirred for 1 hour at 0°C, 1 hour at room temperature, and then 15 minutes later, phosphate buffer (pH 7, 38 ml) was added. The reaction mixture was quenched with MeOH-30% H2O2 (2:1, 100 mL) at <10 °C. After addition and stirring for 20 minutes, water (100 mL) was added and the mixture was concentrated in vacuo. More water (200 mL) was added to the residue and the mixture was extracted with ethyl acetate (3×100 mL). The organic layer was washed with 1N KHSO4 (100 mL), sodium bicarbonate solution (100 mL), and and brine (100 mL), dried over anhydrous Na2SO4, and concentrated in vacuo. The residue was purified by flash column chromatography (10% to 50% ethyl acetate / hexanes). The title compound was obtained as a white solid (7.10 g, 70% yield). 1 H NMR (500 MHz, CDCl3) δ 7.39 (dt, J = 23.4, 7.1 Hz, 3H), 7.30 (d, J = 7.5 Hz, 2H), 5.67 (d, J = 7.1 Hz, 1H), 4.84 - 4.67 (m, 1H), 4.08 - 3.93 (m, 3H), 3.92 - 3.84 (m, 1H), 3.50 (d, J = 9.0 Hz, 1H), 3.24 (d, J = 6.7 Hz, 1H), 2.15 (s, 1H), 1.89 (dd, J = 22.4, 14.8 Hz, 3H), 1.48 (d, J = 21.5 Hz, 9H), 1.33 (d, J = 6.9 Hz, 3H) , 0.88 (d, J = 6.4 Hz, 3H).

[0358] Example 58: (S)-tert-butyl 2-((1R,2R)-1-methoxy-2-methylpropional 3-((4R,5S)-4-methyl-2-oxo-5-phenyloxazolidine- Synthesis of (3-yl)-3-oxopropyl)pyrrolidine-1-carboxylate [ka]

[0359] Under N2 atmosphere, (S)-tert-butyl 2-((1R,2R)-1-hydroxy- 2-Methyl-3-((4R,5S)-4-methyl-2-oxo-5-phenyloxazolidinyl Diazin-3-yl)-3-oxopropyl)pyrrolidine-1-carboxylate (5.1g A mixture of 11.9 mmol) and molecular sieves (4 Å, 5 g) was added to anhydrous dichloroethane (3 0 mL) was added. The mixture was stirred at room temperature for 20 minutes and cooled to 0°C. Hydrogen sponge (6 0.62g, 30.9mmol) was added, followed by trimethyl oxoni tetrafluoroborate. Add 4.40 g (29.7 mmol) of ethanol. Stir at 0°C for 2 hours and at room temperature for 48 hours. The reaction mixture was filtered, and the filtrate was purified by column chromatography (20-70% ethanol). The mixture was concentrated and purified using a solvent (ethyl acetate / hexane) to give the title compound as a white solid (1.80 g, Yield 35%). 1 H NMR (500 MHz, CDCl3) δ 7.46 - 7.27 (m, 5H), 5.65 (s, 1H), 4.69 (s, 1H), 3.92 (s, 1H), 3.83 (s, 1H), 3.48 (s, 3H), 3.17 (s, 2H), 2.02 - 1.68 (m , 5H), 1.48 (d, J = 22.3 Hz, 9H), 1.32 (t, J = 6.0 Hz, 3H), 0.91 - 0.84 (m, 3H).

[0360] Example 59: (2R,3R)-3-((S)-1-(tert-butoxycarbonyl)pyrimidinyl) Synthesis of roridin-2-yl)-3-methoxy-2-methylpropanoic acid [ka]

[0361] (S)-tert-butyl 2-((1R,2R)-1-methoxy-2-methyl-3- ((4R,5S)-4-methyl-2-oxo-5-phenyloxazolidin-3-yl) -3-oxopropyl)pyrrolidine-1-carboxylate (1.80 g, 4.03 mm A solution of 100 ml of HCl in 30 mL of THF and 7.57 mL of HCl was heated at 0°C for 5 min. Add 30% H2O2 (1.44 mL, 14.4 mmol) over 1 min, then add LiOH A solution of (0.27 g, 6.45 mmol) in water (5 mL) was added. Stir at 0° C. for 3 hours. After this, 1N sodium sulfite (15.7 mL) was added and the mixture was allowed to warm to room temperature and stirred overnight. The THF was removed in vacuo and the aqueous phase was washed with dichloromethane (3 x 50 mL) and The oxazolidinone auxiliary was removed. The aqueous phase was acidified to pH 3 with 1N HCl and ethyl acetate was added. The organic layer was washed with brine (50 mL) and The mixture was dried over 1000 kJ / ml, filtered, and concentrated in vacuo to give the title compound as a colorless oil (1.15 g, Yield: 98%. 1 H NMR (500 MHz, CDCl3) δ 3.99 - 3.74 (m, 2H), 3.44 (d, J = 2.6 H z, 3H), 3.23 (s, 1H), 2.60 - 2.45 (m, 1H), 1.92 (tt, J = 56.0, 31.5 Hz, 3H), 1.7 9 - 1.69 (m, 1H), 1.58 - 1.39 (m, 9H), 1.30 - 1.24 (m, 3H).

[0362] Example 60: (4S,5S)-ethyl 4-((tert-butoxycarbonyl)amino Synthesis of )-5-methyl-3-oxohepnoate [ka]

[0363] N-Boc-L-isoleucine (4.55 g, 19.67 mmol) in THF cooled on ice To the solution (20 mL), 1,1'-carbonyldiimidazole (3.51 g, 21.63 m After gas evolution ceased, the resulting mixture was stirred at room temperature for 3.5 hours. Ta.

[0364] A newly prepared solution of isopropylmagnesium bromide in THF (123 mmol, 3 0 mL) with sodium hydrogen malonate (6.50 g, The mixture was stirred at room temperature for 1.5 hours. The magnesium enolate solution was then cooled in an ice-water bath, followed by heating at 0°C. The imidazolide solution was added stepwise via a heavy-ended needle. The reaction mixture was incubated at 0° C. for 30 min. The reaction mixture was stirred at room temperature for 64 hours. 10% aqueous citric acid solution (5 mL) was added to the reaction mixture. The mixture was quenched by the addition of 10% aqueous citric acid (110 mL) and then acidified to pH 3. The mixture was extracted with ethyl acetate (3 x 150 mL). The organic extract was washed with water (50 mL), saturated Sodium bicarbonate solution (50 mL) and saturated sodium chloride solution (50 mL) The extract was washed, dried over Na2SO4, and concentrated in vacuo. The residue was purified by column chromatography on silica gel as eluent to give the title compound. The product was obtained (5.50 g, yield 93%). 1 H NMR (500 MHz, CDCl3) δ 5.04 (d, J = 7.8 Hz, 1H), 4.20 (p, J = 7.0 Hz, 3H), 3.52 (t, J = 10.7 Hz, 2H), 1.96 (d, J = 3.7 H z, 1H), 1.69 (s, 2H), 1.44 (s, 9H), 1.28 (dd, J = 7.1, 2.9 Hz, 3H), 0.98 (t, J = 6.9 Hz, 3H), 0.92 - 0.86 (m, 3H).

[0365] Example 61: (3R,4S,5S)-ethyl 4-((tert-butoxycarbonyl) Synthesis of amino-3-hydroxy-5-methylheptanoate [ka]

[0366] At -60°C, (4S,5S)-ethyl 4-((tert-butoxycarbonyl)amine Ethyl)-5-methyl-3-oxoheptoate (5.90 g, 19.83 mmol) To the alcohol solution (6 mL), borohydride (3.77 g, 99.2 mmol) was added in one portion. The reaction mixture was stirred at -55°C for 5.5 hours and then quenched with 10% aqueous citric acid (100 mL). The resulting solution was acidified to pH 2 by adding 10% aqueous citric acid, followed by acetic acid. The organic extract was extracted with saturated aqueous sodium chloride (10 The residue was washed with 100 mL of ethyl acetate, dried over Na2SO4, and concentrated in vacuo. Purify by column chromatography (10-50% ethyl acetate / hexane) to obtain the target compound. diastereomer (2.20 g, 37% yield) and a mixture of two diastereomers (2.0 g, 37% yield) 34%, approximately 9:1 ratio). 1 H NMR (500 MHz, CDCl3) δ 4.41 (d, J = 9.3 Hz, 1H), 4.17 (tt, J = 7.1, 3.6 Hz, 2H), 4.00 (t, J = 6.9 Hz, 1H), 3.55 (dd, J = 11. 7, 9.3 Hz, 1H), 2.56 - 2.51 (m, 2H), 2.44 (dd, J = 16.4, 9.0 Hz, 1H), 1.79 (d, J = 3.8 Hz, 1H), 1.60 - 1.53 (m, 1H), 1.43 (s, 9H), 1.27 (dd, J = 9.3, 5.0 Hz, 3H ), 1.03 - 0.91 (m, 7H).

[0367] Example 62: (3R,4S,5S)-4-((tert-butoxycarbonyl)amino) Synthesis of 3-hydroxy-5-methylheptanoic acid [ka]

[0368] (3R,4S,5S)-Ethyl 4-((tert-butoxycarbonyl)amino)- 3-Hydroxy-5-methylheptanoate (2.20 g, 7.20 mmol) of ethanol To the solution (22 mL) of 1N sodium hydroxide (7.57 mL, 7.57 mmol) The mixture was stirred at 0° C. for 30 minutes, then at room temperature for 2 hours, and the resulting solution The mixture was acidified to pH 4 by the addition of 1N aqueous hydrochloric acid and extracted with ethyl acetate (3 x 50 mL). The organic extract was washed with 1N aqueous potassium sulfate (50 mL) and saturated sodium chloride. aqueous solution (50 mL), dried over NaSO and concentrated in vacuo to give the title compound Obtained (1.90 g, 95% yield). 1 H NMR (500 MHz, CDCl3) δ 4.50 (d, J = 8.7 Hz, 1H), 4.07 (d, J = 5.5 Hz, 1H), 3.59 (d, J = 8.3 Hz, 1H), 2.56 - 2.45 (m, 2H), 1. 76 - 1.65 (m, 1H), 1.56 (d, J = 7.1 Hz, 1H), 1.45 (s, 9H), 1.26 (t, J = 7.1 Hz, 3H), 0.93 (dd, J = 14.4, 7.1 Hz, 6H).

[0369] Example 63: (3R,4S,5S)-4-((tert-butoxycarbonyl)(methyl Synthesis of )amino)-3-methoxy-5-methylheptanoic acid [ka]

[0370] At 0°C, (3R,4S,5S)-4-((tert-butoxycarbonyl)amino)- A solution of 3-hydroxy-5-methylheptanoic acid (1.90 g, 6.9 mmol) in THF ( Sodium hydride (60% oil suspension, 1.93 g, 48.3 mmol) was added to 40 mL of After stirring for 1 hour, methyl iodide (6.6 mL, 103.5 mmol) was added. After stirring at 0°C for 40 hours, saturated aqueous sodium bicarbonate solution (50 mL) was added. Water (100 mL) was then added, and the mixture was washed with diethyl ether (2×50 mL). The aqueous layer was acidified to pH 3 with 1N aqueous potassium hydrogen sulfate, and then extracted with ethyl acetate (3×50 The combined organic extracts were washed with 5% aqueous sodium thiosulfate (50 mL) and Wash with saturated aqueous sodium chloride (50 mL), dry over Na2SO4, and evaporate in vacuo The mixture was concentrated under reduced pressure to give the title compound (1.00 g, 48% yield). 1 H NMR (500 MHz, CDCl3) δ 3.95 (d, J = 75.4 Hz, 2H), 3.42 (d, J = 4.4 Hz, 3H), 2....

Claims

1. A cell-binding agent-drug conjugate compound of formula (III), (IV), (V), or (VI): 【Chemistry 13-1】 (In the formula, m 1 is 1 or 2, and m 2 and m 3 are 1.) 【Chemistry 13-2】 (wherein m 1 , m 3 and m 4 are 1; m 2 and m 5 are 1 or 2). 【Chemistry 13-3】 (In the formula, m 1 , m 4 , m 5 , m 4 ', m 5 ', m 4 '', and m 5 '' are 1; m 2 and m 3 are 1 or 2.) 【Chemistry 13-4】 (In the formula, m 1 , m 1 ', m 1 '', m 2 , m 2 ', m 2 '', and m 4 ' are 1; m 3 and m 5 are 1 or 2.) During the ceremony, n is 1 to 20; T is CH ２ , alkylcarbonyl, or the following structure: 【Chemistry 9】 During the ceremony, 【Chemistry 10】 is the linking site; Cb, Cb', Cb'' are the same or different and represent cell binding agents / molecules or immunotherapeutic proteins; Drug, Drug', and Drug'' represent the same or different, a cytotoxic agent, or a therapeutic agent, or an immunotherapeutic protein, an antibody or an antibody fragment, or an siRNA, miRNA, mRNA, piRNA, or DNA, or a functional molecule for promoting binding or stabilization of a cell binding agent, or a cell surface receptor binding ligand; 【Chemistry 14】 represents a single bond or a double bond; X １ , X １’ , and X １’’ is NH, and further, X １ , X １’ , and X １’’ may each independently be absent; R １ , R １’ , and R １’’ is absent, the same or different and is alkylcarbonyl or amido; L １ , L １’ , L １’’ , L ２ , L ２’ , and L ２’’ are the same or different and are non-self-immolative moieties or peptide units (1-8 natural or non-natural amino acids) selected from one of the following structures: 【Chemistry 12】 In the formula, ( ＊ ) is the point of attachment to an additional spacer or releasable linker unit, a cytotoxic agent, and / or a cell adhesion molecule (CBA); r is 1-100; m and n are each independently 0-6; Or, L １ , L ２ , and X １ may not be present independently.

2. 2. The conjugate compound of claim 1, wherein said Drug, Drug', or Drug'' is each independently selected from the following: (1) A chemotherapeutic agent selected from the group consisting of: a) alkylating agents selected from the group consisting of: nitrogen mustards: chlorambucil, chlornaphazine, cyclophosphamide, dacarbazine, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, mannomustine, mitobronitol, melphalan, mitolactol, pipobroman, nobembitine, phenesterine, prednimustine, thiotepa, trofosfamide, uracil mustard; CC-1065 (including synthetic analogs of adozelesin, carzelesin, and bizeresin); duocarmycins (including synthetic analogs, KW-2189, and CBI-TMI); benzodiazepine dimers (including pyrrolobenzodiazepines (PBD) or tomaymycin, indolinobe dimers of zodiazepines, imidazobenzothiadiazepines, or oxazolidinobenzodiazepines; nitrosourea compounds: (carmustine, lomustine, chlorozotocin, fotemustine, nimustine, ranimustine); alkylsulfonates: (busulfan, treosulfan, improsulfan, and piposulfan); triazenes (dacarbazine); platinum-containing compounds: (carboplatin, cisplatin, oxaliplatin); aziridines, benzodopa, carboquone, meturedopa, and uredopa; ethylenimines, and methylamelanamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramine, and trimethylolmelamine; b) Plant alkaloids selected from the group consisting of: Vinca alkaloids: (vincristine, vinblastine, vindesine, vinorelbine, navelbine); Taxoids: (paclitaxel, docetaxel) and their analogues, maytansinoids (DM1, DM2, DM3, DM4, DM5, DM6, DM7, maytansine, ansamitocins) and their analogues, cryptophycins (selected from cryptophycin 1 and cryptophycin 8); epothilones, eleutherobin, discodermolides, bryostatin, dolostatin, auristatin, tubulysin, cephalostatin; pancratistatin; sarcodictiin; spongistatin; c) DNA topoisomerase inhibitors selected from the group consisting of: [epipodophyllins: (9-aminocamptothecin, camptothecin, crisnatol, daunomycin, etoposide, etoposide phosphate, irinotecan, mitoxantrone, novantrone, retinoic acid (retinol), teniposide, topotecan, 9-nitrocamptothecin (RFS 2000); mitomycin: (mitomycin C)]; d) Antimetabolites selected from the group consisting of: {[Antifolates: Dihydrofolate reductase inhibitors: (methotrexate, trimetrexate, denopterin, pteropterin, aminopterin (4-aminopteroic acid), or folic acid analogs); IMP dehydrogenase inhibitors (mycophenolic acid, tiazofurin, ribavirin, EICAR); Ribonucleotide reductase inhibitors (hydroxyurea, deferoxamine)]; [Pyrimidine analogs: Uracil analogs (anthraquinone, uracil analogs); Cytabine, azacitidine, 6-azauridine, capecitabine (Xeloda), carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, 5-fluorouracil, floxuridine, raltitrexed (Tomudex)); cytosine analogues: (cytarabine, cytosine arabinoside, fludarabine); purine analogues: (azathioprine, fludarabine, mercaptopurine, thiamiprine, thioguanine)]; folic acid supplements, fluorescein acid}; e) Hormonal therapy agents selected from the following group: {Receptor antagonists: [Antiestrogen: (megestrol, raloxifene, tamoxifen); LHRH agonists: (goserelin, leuprolide acetate); Antiandrogens: (bicalutamide, flutamide, calsterone, dromostanolone propionate, epithiostanol, goserelin, leuprolide, mepitiostane, nilutamide, testolactone, trilostane, and other androgen inhibitors)]; retinoids: [vitamin D3 analogs: (CB1093, EB1089, KH1060, cholecalciferol, ergocalciferol); photodynamic therapy agents: (verteporfin, phthalocyanines, photosensitizer Pc4, demethoxy-hypocrelin A); cytokines: (interferon alpha, interferon gamma, tumor necrosis factor (TNF), TNF domain-containing human proteins)]}; f) Kinase inhibitors selected from the group consisting of BIBW2992 (anti-EGFR / Erb2), imatinib, gefitinib, pegaptanib, sorafenib, dasatinib, sunitinib, erlotinib, nilotinib, lapatinib, axitinib, pazopanib, vandetanib, E7080 (anti-VEGFR2), mubritinib, ponase tinib (AP24534), bafetinib (INNO-406), bosutinib (SKI-606), cabozantinib, vismodegib, iniparib, ruxolitinib, CYT387, axitinib, tivozanib, sorafenib, bevacizumab, cetuximab, trastuzumab, ranibizumab, panitumumab, ispinesib; g) a poly(ADP-ribose)polymerase (PARP) inhibitor selected from the group consisting of olaparib, niraparib, iniparib, talazoparib, veliparib, CEP9722 (Cephalon), E7016 (Eisai), BGB-290 (BeiGene), or 3-aminobenzamide; h) Antibiotics selected from the group consisting of enediyne antibiotics (calicheamicin, calicheamicin gamma 1, delta 1, alpha 1, and beta 1; dynemicins including dynemicin A and deoxydynemicin; esperamicin, kedarcidin, C-1027, maduropeptin, and neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomycin, actinomycin, anthramycin, azaserine, bleomycin, cactinomycin, carabicin, carminomycin, carzinov chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid, nogalamycin, olivomycin, peplomycin, potfilomycin, puromycin, queramycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; i) Polyketides (acetogenins), bullatacin and bullatacinone; gemcitabine, epoxomicin (including carfilzomib), bortezomib, thalidomide, lenalidomide, pomalidomide, tosedostat, zybrestat, PLX4032, STA-9090, Stimuvax, allovectin-7, zygeva, probenge, elvoy, isoprenylation inhibitors (selected from lovastatin), dopaminergic neurotoxins (selected from 1-methyl-4-phenylpyridine ion, actinomycin (selected from the group consisting of actinomycin D, dactinomycin), bleomycin (selected from the group consisting of bleomycin A2, bleomycin B2, peplomycin), anthracycline (selected from the group consisting of daunorubicin, doxorubicin (adriamycin), idarubicin, epirubicin, pirarubicin, zorubicin), mitoxantrone, MDR inhibitors or verapamil, Ca ２＋ ATP inhibitors or thapsigargin, histone deacetylase inhibitors (selected from the group consisting of vorinostat, romidepsin, panobinostat, valproic acid, mocetinostat (MGCD0103), belinostat, PCI-24781, entinostat, SB939, resminostat, gibinostat, AR-42, CUDC-101, sulforaphane, trichostatin A); thapsigargin, celecoxib, glitazones, epigallocatechin gallate, disulfiram, salinosporamide A, antiadrenal drugs (selected from the group consisting of aminoglutethimide, mitotane, trilostane); aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; arabinoside, bestravcil; bisantrene; edat Rexart; Defofamine, Demecolcine, Diaziquone, Elfornithine (DFMO), Elfomicin; Elliptinium acetate, Etocurside; Gallium nitrate; Gacytosine; Hydroxyurea; Ibandronate, Lentinan; Lonidamine; Mitoguazone; Mitoxatrone; Mopidamol; Nitraerin; Pentostatin; Fenamet; Pirarubicin; Podophyllic acid; 2-Ethylhydrazide; Procarbazine; PSK (registered trademark); Razoxane; Rhizoxin; Sizofiran; Spirogermanium; Tenuazonic acid; Triaziquone; 2,2',2''-Trichlorotriethylamine; Trichothecenes (selected from the group consisting of T2 toxin, verrucarin A, roridin A, and anguidine); Urethane, siRNA, antisense medicines; (2) Anti-autoimmune disease drugs: cyclosporine, cyclosporine A, aminocaproic acid, azathioprine, bromocriptine, chlorambucil, chloroquine, cyclophosphamide, corticosteroids (selected from the group consisting of amcinonide, betamethasone, budesonide, hydrocortisone, flunisolide, fluticasone propionate, fluocortodanazol, dexamethasone, triamcinolone acetonide, beclomethasone dipropionate), DHAE, etanercept, hydroxychloroquine, infliximab, meloxicam, methotrexate, mycophenolate mofetil, prednisone, sirolimus, tacrolimus; (3) Anti-infective drugs: a) Aminoglycosides: amikacin, astromycin, gentamicin (netimicin, sisomicin, isepamicin), hygromycin B, kanamycin (amikacin, arbekacin, bekanamycin, dibekacin, tobramycin), neomycin (framycetin, paromomycin, ribostamycin), netilmicin, spectinomycin, streptomycin, tobramycin, verdamycin; b) Amphenicols: Azidamphenicol, Chloramphenicol, Florfenicol, Thiamphenicol; c) Ansamycins: geldanamycin, herbimycin; d) Carbapenems: biapenem, doripenem, ertapenem, imipenem / cilastatin, meropenem, panipenem; e) Cephalosporins: carbacephem (loracarbef), cephacetrile, cefachlor, cephradine, cefadroxil, cephalonium, cephaloridine, cephalothin or cephalosporin, cephalexin, cephaloglycin, cefamandole, cephapirin, cefatrizine, cefazaflur, cefazedone, cefazolin, cefbuperazone, cefcapene, cefdaloxime, cefepime, cefminox, cefoxitin, cefprozil, cephalosporin, ceftezol, cefuroxime, cefixime, cefdinir, cefditoren, cefepi cefetamet, cefmenoxime, cefodizime, cefonicid, cefoperazone, ceforanide, cefotaxime, cefotiam, cefozopran, cephalexin, cefpimizole, cefpiramide, cefpirome, cefpodoxime, cefprozil, cefquinome, cefsulodin, ceftazidime, cefteram, ceftibuten, ceftiolene, ceftizoxime, ceftobiprole, ceftriaxone, cefuroxime, cefuzonam, cephamycins (cefoxitin, cefotetan, cefmetazole), oxacephems (flomoxef, latamoxef); f) Glycopeptides: bleomycin, vancomycin (oritavancin, telavancin), teicoplanin (dalbavancin), ramoplanin; g) Glycylcyclines: Tigecycline; h) β-lactamase inhibitors: penams (sulbactam, tazobactam), clavams (clavulanic acid); i) Lincosamides: Clindamycin, Lincomycin; j) Lipopeptides: daptomycin, A54145, calcium-dependent antibiotic (CDA); k) Macrolides: azithromycin, cethromycin, clarithromycin, dirithromycin, erythromycin, flurithromycin, josamycin, ketolides (telithromycin, cethromycin), midecamycin, miocamycin, oleandomycin, rifamycin (rifampicin, rifampin, rifabutin, rifapentine), rokitamycin, roxithromycin, spectinomycin, spiramycin, tacrolimus (FK506), troleandomycin, telithromycin; l) Monobactams: aztreonam, tigemonam; m) Oxazolidinone: Linezolid; n) Penicillins: amoxicillin, ampicillin (pivanpicillin, hetacillin, bacampicillin, methampicillin, talampicillin), azidocillin, azlocillin, benzylpenicillin, benzathine benzylpenicillin, benzathine phenoxymethylpenicillin, clometocillin, procaine benzylpenicillin, carbenicillin (carindacillin), cloxacillin, dicloxacillin, epicillin, flucloxacillin, mecillinam (pivmecillinam), mezlocillin, methicillin, nafcillin, oxacillin, penamecillin, penicillin, phenethicillin, phenoxymethylpenicillin, piperacillin, propicillin, sulbenicillin, temocillin, ticarcillin; o) Polypeptides: bacitracin, colistin, polymyxin B; p) Quinolones: alatrofloxacin, balofloxacin, ciprofloxacin, clinafloxacin, danofloxacin, difloxacin, enoxacin, enrofloxacin, floxin, garenoxacin, gatifloxacin, gemifloxacin, grepafloxacin, canotrovafloxacin, levofloxacin, lomefloxacin, marbofloxacin, moxifloxacin, nadifloxacin, norfloxacin, orbifloxacin, ofloxacin, pefloxacin, trovafloxacin, grepafloxacin, sitafloxacin, sparfloxacin, temafloxacin, tosufloxacin, trovafloxacin; q) Streptogramins: Pristinamycin, Quinupristin / Dalfopristin; r) Sulfonamides: mafenide, prontosil, sulfacetamide, sulfamethizole, sulfanilamide, sulfasalazine, sulfisoxazole, trimethoprim, trimethoprim-sulfamethoxazole (cotrimoxazole); s) steroidal antibacterial agents: selected from fusidic acid; t) Tetracyclines: doxycycline, chlortetracycline, clomocycline, demeclocycline, lymecycline, meclocycline, methacycline, minocycline, oxytetracycline, penimepicycline, rolitetracycline, tetracycline, glycylcycline (selected from tigecycline); u) Other types of antibiotics: annonacin, arsphenamine, bactoprenol inhibitors (bacitracin), DADAL / AR inhibitors (cycloserine), dictyostatin, discodermolide, eleutherobin, epothilone, ethambutol, etoposide, faropenem, fusidic acid, furazolidone, isoniazid, laulimalide, metronidazole, mupirocin, mycolactone, NAM synthesis inhibitors (fosfomycin), nitrofurantoin, paclitaxel, platensimycin, pyrazinamide, quinupristin / dalfopristin, rifampin (rifampin), tazobactam tinidazole, uvaricin; (4) Antiviral drugs: a) Entry / fusion inhibitors: aplaviroc, maraviroc, vicriviroc, gp41 (enfuvirtide), PRO140, CD4 (ibalizumab); b) Integrase inhibitors: raltegravir, elvitegravir, globoidnan A; c) Maturation inhibitors: Bevirimat, Vivicon; d) Neuraminidase inhibitors: oseltamivir, zanamivir, peramivir; e) Nucleosides and Nucleotides: Abacavir, Acyclovir, Adefovir, Amdoxovir, Apricitabine, Brivudine, Cidofovir, Clevudine, Dexerbucitabine, Didanosine (DDI), Elvucitabine, Emtricitabine (FTC), Entecavir, Famciclovir, Fluorouracil (5-FU), 3'-Fluoro-substituted 2',3'-deoxynucleoside analogues (3'-Fluoro-2',3'-dideoxythymidine (FLT) and 3'-Fluoro-2',3'-dideoxy guanosine (FLG), fomivirsen, ganciclovir, idoxuridine, lamivudine (3TC), L-nucleosides (β-L-thymidine and β-L-2'-deoxycytidine), penciclovir, rasvir, ribavirin, stampidine, stavudine set (d4T), taribavirin (viramidine), telbivudine, tenofovir, trifluridine, valacyclovir, valganciclovir, zalcitabine (ddC), zidovudine (AZT); f) Non-nucleosides: amantadine, ateviridine, capravirine, diarylpyrimidines (etravirine, rilpivirine), delavirdine, docosanol, emivirine, efavirenz, foscarnet (phosphorylformate), imiquimod, interferon alpha, loviride, rhodenosine, methisazone, nevirapine, NOV-205, peginterferon alpha, podophyllotoxin, rifampicin, rimantadine, resiquimod (R-848), tromantadine; g) Protease inhibitors: amprenavir, atazanavir, boceprevir, darunavir, fosamprenavir, indinavir, lopinavir, nelfinavir, pleconaril, ritonavir, saquinavir, telaprevir (VX-950), tipranavir; h) Other antivirals: abzyme, arbidol, calanolide A, ceragenin, cyanovirin-N, diarylpyrimidine, epigallocatechin gallate (EGCG), foscarnet, griffithsin, taribavirin (viramidine), hydroxyurea, KP-1461, miltefosine, pleconaril, portmanteau inhibitors, ribavirin, seliciclib; (5) ３ H. １１ C. １４ C. １８ F. ３２ P. ３５ S. ６４ Cu, ６８ G.A. ８６ Y. ９９ Tc, １１１ In, １２３ I, １２４ I, １２５ I, １３１ I, １３３ Xe, １７７ Lu, ２１１ At, or ２１３ A radioisotope (radionuclide) that may be selected from Bi; (6) Chromophore molecules capable of absorbing certain light such as UV light, fluorescent light, IR light, near-IR light, visual light; xanthophores, erythrophores, iridophores, leucophores, melanophores, cereus, and the class or subclass of phosphors, which are fluorescent chemicals that re-emit light with light, visual light sensitive molecules, light emitting molecules, luminescent molecules, luciferin compounds, classes or subclasses of visible light transmitting molecules, light emitting molecules, luminescent molecules, luciferin compounds; non-protein organic phosphors such as: xanthene derivatives (fluorescein, rhodamine, , Oregon Green, Eosin, and Texas Red); cyanine derivatives (cyanine, indocarbocyanine, oxacarbocyanine, thiacarbocyanine, and merocyanine); squalene derivatives and ring-substituted squaraines (including Seta, SeTau, and Square dyes); naphthalene derivatives (dansyl and prodan derivatives); coumarin derivatives; oxadiazole derivatives (pyridyloxazole, nitrobenzoxadiazole, and benzoxadiazole); anthracene derivatives (anthraquinones including DRAQ5, DRAQ7, and CyTRAK Orange); pyrene derivatives (cascade blue); oxazine derivatives (Nile red, Nile blue, cresyl violet, oxazine 170); acridine derivatives (proflavine, acridine orange, acridine yellow, etc.). Arylmethine derivatives (auramine, crystal violet, malachite green). Tetrapyrrole derivatives (porphine, phthalocyanine, bilirubin); the following fluorophore compounds: CF dyes (Biotium), DRAQ and CyTRAK probes (Bio-Status), BODIPY (Invitrogen), Alexa Fluor (Invitrogen), DyLight Fluor (Thermo Scientific, Pierce), Atto and Trancy (Sigma Aldrich), FluoProbes (Interchim), Abberior dyes (Abberior), DY and MegaStokes dyes (Dyomics), SulfoCy dyes (Cyandye), HiLyte Fluor (AnaSpec), Seta, SeTau and Square dyes (SETA BioMedicals), Quasar and Cal Flour dyes (Biosearch Technologies), SureLight dyes (APC, RPEPerCP, phycobilisome) (Columbia Biosciences), APC, APCXL, RPE, BPE (Phyco-Biotech), allophycocyanin (APC), aminocoumarin, APC-Cy7 conjugate, BODIPY-FL, Cascade Blue, Cy2, Cy3, Cy3.5, Cy3B, Cy5, Cy5.5, Cy7, fluorescein, FluorX, hydroxycoumarin, Lissamine rhodamine B, Lucifer Yellow, methoxycoumarin, NBD, Pacific Blue, PacificOrange, PE-Cy5 conjugate, PE-Cy7 conjugate, PerCP, R-Phycoerythrin (PE), Red613, Seta-555-azide, Seta-555-DBCO, Seta-555-NHS, Seta-580-NHS, Seta-680-NHS, Seta-780-NHS, Seta-APC-780, Seta-PerCP-680, Seta-R-PE-670, SeTau380-NHS, SeTau405-maleimide, SeTau405-NHS, SeTau425-NHS, SeTau647-NHS, Texas Red, TRITC, TruRed , X-rhodamine, 7-AAD (7-aminoactinomycin D, CG selective), acridine orange, chromomycin A3, CyTRAK orange (Biostatus, red excitation dark), DAPI, DRAQ5, DRAQ7, ethidium bromide, Hoechst 33258, Hoechst 33342, LDS751, mithramycin, propidium iodide (PI), SYTOX blue, SYTOX green, SYTOX orange, thiazole orange, TO-PRO: cyanine monomer, TOTO-1, TO-PRO-1, TOTO-3, TO-PRO-3, YOSETA-1, YOYO-1. Fluorescent dyes that can be linked to the conjugates of the invention for cell studies are selected from the following compounds or their derivatives: DCFH (2'7'dichlorodihydro-fluorescein, oxidized form), DHR (dihydrorhodamine 123, oxidized form), Fluo-3 (AM ester, pH>6), Fluo-4 (AM ester, pH 7.2), Indo-1 (AM ester, low / high calcium (Ca ２＋ )), SNARF (pH 6 / 9), allophycocyanin (APC), AmCyan1 (tetramer, Clontech), AsRed2 (tetramer, Clontech), Azami Green (monomer, MBL), azurite, B-phycoerythrin (BPE), cerulean, CyPet, DsRed monomer (Clontech), DsRed2 ("RFP", Clontech), EBFP, EBFP2, ECFP, EGFP (weak dimer, Clontech), Emerald (weak dimer, Invitrogen), EYFP (weak dimer, Clontech), GFP (S65A mutant), GFP (S65C mutant), GFP (S65L mutant), GFP (S65T mutant), GFP (Y66F mutant), GFP (Y66H mutant), GFP (Y66W mutant), GFP ｕｖ , HcRed1, J-Red, Katyusha, Kusabira Orange (monomer, MBL), mCFP, mCherry, mCitrine, Midriishi Cyan (dimer, MBL), mKate (TagFP635, monomer, Evrogen), mKeima-Red (monomer, MBL), mKO, mOrange, mPlum, mRaspberry, mRFP1 (monomer, Tsien Lab), mStrawberry, mTFP1, mTurquoise2, P3 (phycobilisome complex), Peridinin Chlorophyll (PerCP), R-phycoerythrin (RPE), T-Sapphire, TagCFP (dimer, Evrogen), TagGFP (dimer, Evrogen), TagRFP (dimer, Evrogen), TagYFP (dimer, Evrogen), tdTomato (tandem dimer), Topaz, TurboFP602 (dimer , Evrogen), TurboFP635 (dimer, Evrogen), TurboGFP (dimer, Evrogen), TurboRFP (dimer, Evrogen), TurboYFP635 (dimer, Evrogen), Venus, native GFP, YPet, Zs Green 1 (tetramer, Clontech), Zs Yellow 1 (tetramer, Clontech). (7) Cell-binding ligands or receptor agonists that can be selected from the following: folic acid derivatives; glutamic acid urea derivatives; somatostatin and its analogs (selected from the group consisting of octreotide (Sandostatin) and lanreotide (Somatrin); aromatic sulfonamides; pituitary adenylate cyclase activating peptide (PACAP) (PAC1); vasoactive intestinal peptide (VIP / PACAP) (VPAC1, VPAC2); melanocyte stimulating hormone (α-MSH); cholecystokinin (CCK) / gastrin receptor agonists; bombesin (Pyr-Gln-Arg-Leu-Gly-Asn-Gln-Trp-Ala-Val-Gly-His-Leu-Met-NH 2 Neurotensin receptor ligands (NTR1, NTR2, NTR3); substance P (NK1 receptor) ligands; neuropeptide Y (Y1-Y6); homing peptides including RGD (Arg-Gly-Asp), NGR (Asn-Gly-Arg), dimeric and multimeric cyclic RGD peptides (cRGDfVc), TAASGVRSMH and LTLRWVGLMS (chondroitin sulfate proteoglycan NG2 receptor ligand) and F3 peptide; cell-permeable peptides tides (CPPs); buserelin (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-Pro-NHEt), gonadorelin (Pyr-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NHEt), which act by targeting follicle-stimulating hormone (FSH) and luteinizing hormone (LH), as well as testosterone production, selected from the group consisting of luteinizing hormone releasing hormone (LHRH) agonists and antagonists, and gonadotropin releasing hormone (GnRH) agonists. 2 ), goserelin (Pyr-His-Trp-Ser-Tyr-D-Ser(OtBu)-Leu-Arg-Pro-AzGly-NH2), histrelin (Pyr-His-Trp-Ser-Tyr-D-His(N-benzyl)-Leu-Arg-Pro-NHEt), leuprolide (Pyr-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro-NHEt), nafrelin (Pyr-His-Trp-Ser-Tyr-2Nal-Leu-Arg-Pro-Gly-NH 2 ), Triptorelin (Pyr-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH 2 ), nafarelin, deslorelin, abarelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-(N-Me)Tyr-D-Asn-Leu-isopropyl-Pro-DAla-NH 2 ), Cetrorelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-Cit-Leu-Arg-Pro-D-Ala-NH 2 ), Degarelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-4-amioPhe(L-hydroorotyl)-D-4-amioPhe(carbamoyl)-Leu-isopropylLys-Pro-D-Ala-NH 2 ), and ganirelix (Ac-D-2Nal-D-4-chloroPhe-D-3-(3-pyridyl)Ala-Ser-Tyr-D-(N9, N10-diethyl)-homoArg-Leu-(N9, N10-diethyl)-homoArg-Pro-D-Ala-NH 2 ); pattern recognition receptors (PRRs) selected from the group consisting of Toll-like receptor (TLR) ligands, C-type lectins, and Nod-like receptor (NLRs) ligands; calcitonin receptor agonists; integrin receptors and their receptor subtypes (α Ｖ β １ , α Ｖ β ３ , α Ｖ β ５ , α Ｖ β ６ , α ６ β ４ , α ７ β １ , α Ｌ β ２ , α ＩＩｂ β ３ ) agonists (GRGDSPK, cyclo(RGDfV)(L1) and its derivatives [cyclo(-N(Me)R-GDfV), cyclo(R-Sar-DfV), cyclo(RG-N(Me)D-fV), cyclo(RGD-N(Me)f-V), cyclo(RGDf-N(Me)V-)(ciliates)]; nanobodies (VHH (camel Ig) derivatives); domain antibodies (dAb, VH or VL domain derivatives), bispecific T cell induction (Bite, bispecific antibodies); dual affinity retargeting (DART, bispecific antibodies); tetravalent tandem antibodies (TandAb, dimerized bispecific antibodies); avimers; EGF receptor and VEGF receptor agonists; (8) Pharmaceutically acceptable salts, acids, or derivatives of any of the above drugs.

3. 2. The conjugate compound of claim 1, wherein the Drug, Drug', or Drug'' is a chromophore molecule.

4. 2. The conjugate compound of claim 1, wherein one, two or all of said Drug, Drug' or Drug'' are each independently poly(ethylene glycols) (PEGs), poly(propylene glycols), copolymers of ethylene oxide or propylene oxide having a molecular weight of 44 Da to 300 kDa, or analogs thereof.

5. 2. The conjugate compound of claim 1, wherein one, two or all of said Drug, Drug' or Drug'' are each independently a cell binding ligand or a cell receptor agonist, or a cell receptor binding molecule.

6. The Drug, Drug', or Drug'' is selected from the group consisting of tubulysin, calicheamicin, auristatin, maytansinoids, CC-1065, daunorubicin and doxorubicin compounds, taxanoids, cryptophycins, epothilones, benzodiazepine dimers (selected from the group consisting of pyrrolobenzodiazepines (PBD), tomaymycin, anthramycin, indolinobenzodiazepines, imidazobenzothiadiazepines, or oxazolidinobenzodiazepine dimers), calicheamicin and enediyne antibiotics, actinomycin, amanitin, azaserine, bleomycin, epirubicin, tamoxifen, idarubicin, dolastatin / auristatin (monomethylauristatin E, MMAE, MMAF, auristatin PYE, auristatin TP, auristatin 2- AQ, 6-AQ, EB (AEB), and EFP (AEFP), duocarmycins, geldanamycins, methotrexate, thiotepa, vindesines, vincristine, hemiasterin, nazumamides, microginins, radiosumins, alterobactins, microsclerodermins, theonellamides, esperamicins, siRNA, miRNA, piRNA, nucleases, and / or pharmaceutically acceptable salts of any of the above molecules.

7. 2. The conjugate compound of claim 1, wherein the cell-binding agent / molecule is selected from antibodies, proteins, vitamins, peptides, polymeric micelles, liposomes, lipoprotein-based drug carriers, nanoparticle drug carriers, dendrimers, and molecules coated with cell-binding ligands, or combinations thereof.

8. The Cb, Cb', or Cb'' may independently be an antibody, an antibody-like protein, a full-length antibody (selected from a polyclonal antibody, a monoclonal antibody, a dimer, a multimer, a multispecific antibody (selected from a bispecific antibody or a trispecific antibody)); a single chain antibody, an antibody fragment that binds to a target cell, a monoclonal antibody, a single chain monoclonal antibody, a monoclonal antibody fragment that binds to a target cell, a chimeric antibody, a chimeric antibody fragment that binds to a target cell, a domain antibody, a domain antibody fragment that binds to a target cell, a resurfaced antibody, a single chain resurfaced antibody, a resurfaced antibody that binds to a target cell, 6. The conjugate compound of claim 1 or 5, selected from the group consisting of shaped antibody fragments, probodies, humanized antibodies, resurfaced humanized antibodies, single chain humanized antibodies, humanized antibody fragments that bind to target cells, anti-idiotypic (anti-Id) antibodies, CDRs, dimers, trimers, tetramers, miniantibodies, probodies, small immune proteins (SIPs), lymphokines, hormones, vitamins, growth factors, colony stimulating factors, nutrient transport molecules, large molecular weight proteins, and nanoparticles or polymers modified with antibodies or large molecular weight proteins.

9. The conjugate compound of claim 1 or 7, wherein the Cb, Cb', or Cb'' is a molecule or agent capable of targeting tumor cells, virus-infected cells, microbial-infected cells, parasite-infected cells, autoimmune disease cells, activated tumor cells, bone marrow cells, activated T cells, affected B cells, or melanocytes, or cells expressing one or more of the following antigens or receptors: CD3, CD4, CD5, CD6, CD7, CD8, CD9, CD10, CD11a, CD11b, CD11c, CD12w, CD14, CD15, CD16, CDw17, CD18. , CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD26, CD27, CD28, CD29, CD30, CD31, CD32, CD33, CD34, CD35, CD36, CD37, CD38, CD39, CD40, CD41, CD42, CD 43, CD44, CD45, CD46, CD47, CD48, CD49b, CD49c, CD51, CD52, CD53, CD54, CD 55, CD56, CD58, CD59, CD61, CD62E, CD62L, CD62P, CD63, CD66, CD68, CD69, C D70, CD72, CD74, CD79, CD79a, CD79b, CD80, CD81, CD82, CD83, CD86, CD87, CD88, CD89, CD90, CD91, CD95, CD96, CD98, CD100, CD103, CD105, CD106, CD1 09, CD117, CD120, CD123, CD125, CD126, CD127, CD133, CD134, CD135, CD137 , CD138, CD141, CD142, CD143, CD144, CD147, CD151, CD147, CD152, CD154, C D156, CD158, CD163, CD166, CD168, CD174, CD180, CD184, CDw186, CD194, CD195, CD200, CD200a, CD200b, CD209, CD221, CD227, CD235a, CD240, CD262, CD271, CD274, CD276 (B7-H3), CD303, CD304, CD309, CD326, 4-1BB, 5AC, 5T4 (trophoblast glycoprotein, TPBG, 5T4, Wnt activation inhibitor 1 or WAIF1), adenocarcinoma antigen, AGS-5, AGS-22M6,Activin receptor-like kinase 1, AFP, AKAP-4, ALK, α integrin, αvβ6, aminopeptidase N, amyloid beta, androgen receptor, angiopoietin 2, angiopoietin 3, annexin A1, anthrax toxin protective antigen, anti-transferrin receptor, AOC3 (VAP-1), B7-H3, anthrax, BAFF (B-cell activating factor), BCMA, B-lymphoma cells, bcr-abl, bombesin, BORIS, C5, C242 antigen, CA125 (carbohydrate antigen 125, MUC16), CA-IX (or CAIX, carbonic anhydrase 9), CALLA, CanAg, canine IL31, carbonic anhydrase IX, cardiac myosin, CCL11 (C-C motif chemokine 11), C CR4 (CC chemokine receptor type 4, CD194), CCR5, CD3E (epsilon), CEA (carcinoembryonic antigen), CEACAM3, CEACAM5 (carcinoembryonic antigen), CFD (factor D), Ch4D5, cholecystokinin 2 (CCK2R), CLDN18 (claudin-18), clumping factor A, cMet, CRIPTO, FCSF1R (colony stimulating factor 1 receptor, CD115), CSF2 (colony stimulating factor 2, granulocyte-macrophage colony-stimulating factor (GM-CSF)), CSP4, CTLA4 (cytotoxic T-lymphocyte-associated protein 4), CTAA16.88 tumor antigen, CXCR4 (CD184), CXC chemokine receptor type 4, cADP-ribose hydrolase, Cyclin B1, CYP1B1, cytomegalovirus, cytomegalovirus glycoprotein B, dabigatran, DLL3 (delta-like ligand 3), DLL4 (delta-like ligand 4), DPP4 (dipeptidyl peptidase 4), DR5 (death receptor 5), Escherichia coli Shiga toxin type 1, Escherichia coli Shiga toxin type 2, ED-B, EGFL7 (EGF-like domain containing protein 7), EGFR, EGFRII, EGFRvIII, E endoglin (CD105), endothelin B receptor, endotoxin, EpCAM (epithelial cell adhesion molecule), EphA2, episialin, ERBB2 (epidermal growth factor receptor 2), ERBB3, ERG (TMPRSS2ETS fusion gene), Escherichia coli, ETV6-AML, FAP (fibroblast activation protein α), FCGR1, α-fetoprotein, fibrin II, β chain, fibronectin ectodomain B,FOLR (folate receptor), folate receptor α, folate hydrolase, respiratory syncytial virus Fos-related antigen 1F protein, Frizzled receptor, fucosyl GM1, GD2 ganglioside, G-28 (cell surface glycolipid antigen), GD3 idiotype, GloboH, glypican 3, N-glycolylneuraminic acid, GM3, GMCSF receptor α chain, growth differentiation factor 8, GP100, GPNMB (transmembrane protein NMB), GUCY2C (guanylate cyclase 2C, guanylate cyclase C (GC-C), intestinal guanylate cyclase, guanylate cyclase PCR-C receptor, heat-stable enterotoxin receptor (hSTAR)), heat shock protein, hemagglutinin, hepatitis B surface antigen, hepatitis B virus, HER1 (human epidermal growth factor receptor 1), HER2, HER2 / neu, HER3 (ERBB-3), IgG4, HGF / SF (stem cell growth factor / scatter factor), HHGFR, HIV-1, histone complex, HLA-DR (human leukocyte antigen), HLA-DR10, HLA-DRB, HMWMAA, human chorionic gonadotropin, HNGF, human cell scatter factor receptor kinase, HPV E6 / E7, Hsp90, hTERT, ICAM-1 (intercellular adhesion molecule 1), idiotype, IGF1R (IGF-1, insulin-like growth factor 1 receptor), IGHE, IFN-γ, influenza hemagglutinin, IgE, IgE Fc region, IGHE, IL-1, IL-2 receptor (interleukin 2 receptor), IL-4, IL-5, IL-6, IL-6R (interleukin 6 receptor), IL-9, IL-10, L-12, IL-13, IL-17, IL-17A, IL-20, IL-22, IL-23, IL-31RA, ILGF2 (insulin-like growth factor 2), integrins (α4, α, ＩＩＩｂ β ３ , αvβ3, α ４ β ７ , α5β1, α6β4, α7β7, αIIβ3, α5β5, αvβ5), interferon-gamma-inducible protein, ITAGA2, ITGB2, KIR2D, Kappa Ig, LCK, Le, legumain, Lewis-Y antigen, LFA-1 (lymphocyte function-associated antigen 1, CD11a), LHRH, LINGO-1, lipoteichoic acid, LIV1A, LMP2, LTA, MAD-CT-1, MAD-CT-2, MAGE-1, MAGE-2, MAGE-3, MAGEA1, MAGEA3, MAGEA4, MART1, MCP-1, MIF (macrophage migration inhibitory factor or glycosylation inhibitory factor (GIF)), MS4A1 (transmembrane 4 domain subfamily A member 1), MSLN (mesothelin), MUC1 (mucin 1, cell surface associated (MUC1) or polymorphic epithelial mucin (PEM), MUC1-KLH, MUC16 (CA125), MCP1 (monocyte chemotactic protein 1), MelanA / MART1, ML-IAP, MPG, MS4A1 (transmembrane 4 domain subfamily A), MYCN, myelin-associated glycoprotein, myostatin, NA17, NARP-1, NCA-90 (granulocyte antigen), Nectin-4 (ASG-22ME), NGF, neuronal apoptosis-regulating proteinase 1, NOGO-A, Notch receptor, nucleolin, Neu oncogene product, NY-BR-1, NY-ESO-1, OX-40, OxLDL (oxidized low density lipoprotein), OY-TES1, P21, p53 non-mutant, P97, Page 4 , PAP, ,Anti-(N-glycolylneuraminic acid) paratope, PAX3, PAX5, PCSK9, PDCD1 (PD-1, programmed cell death protein 1, CD279), PDGF-Rα, (platelet-derived growth factor receptor α), PDGFR-β, PDL-1, PLAC1, PLAP-like testicular alkaline phosphatase, platelet-derived growth factor receptor β, sodium phosphate cotransporter, PMEL17, polysialic acid, proteinase 3 (PR1), prostate cancer, PS (phosphatidylserine), prostate cancer cells, Pseudomonas aeruginosa, PSMA, PSA, PSCA, rabies virus glycoprotein, RHD (Rh polypeptide 1 (RhPI), CD240), Rhesus factor, RANKL, PhoC,Ras mutants, RG55, ROBO4, respiratory syncytial virus, RON, ROR1, sarcoma metastasis breakpoint, SART3, sclerostin, SLAMF7 (SLAM family member 7), selectin P, SDC1 (syndecan 1), sLe(a), somatomedin C, SIP (sphingosine-1-phosphate), somatostatin, sperm protein 17, SSX2, STEAP1 (six transmembrane spanning of the prostate 1) epithelial antigen), STEAP2, STn, TAG-22 (tumor-associated glycoprotein 72), survivin, T cell receptor, T cell transmembrane protein, TEM1 (tumor epithelial marker 1), TENB2, tenascin-C (TN-C), TGF-α, TGF-β (transforming growth factor beta), TGF-β1, TGF-β2 (transforming growth factor beta 2), Tie (CD202b), Tie2, TIM-1 (CDX-014), TN, TNF, TNF-α, TNFRSF8, TNFRSF10B (tumor necrosis factor receptor superfamily member 10B), TNFRSF13B (tumor necrosis factor receptor superfamily member 13B), TPBG (trophoblast glycoprotein), TRAIL-R1 (tumor necrosis-inducing ligand receptor 1), TRAILR2 (death receptor 5 (DR5)), tumor-associated calcium stimulator Signal transducer 2, tumor-specific glycosylation of MUC1, TWEAK receptor, TYRP1 (glycoprotein 75), TRP-2, tyrosinase, VCAM-1 (CD106), VEGF, VEGF-A, VEGF-2 (CD309), VEGFR-1, VEGFR2, or vimentin, WT1, XAGE1, or any insulin growth factor receptor, or any epidermal growth factor receptor.

10. The conjugate compound of claim 9, wherein the tumor cells are selected from lymphoma cells, myeloma cells, renal cells, breast cancer cells, prostate cancer cells, ovarian cancer cells, colon cancer cells, gastric cancer cells, squamous cell carcinoma cells, small cell lung cancer cells, non-small cell lung cancer cells, or testicular cancer cells.

11. The cytotoxic agent is selected from tubulysin, maytansinoids, taxanoids, CC-1065, daunorubicin and doxorubicin compounds, benzodiazepine dimers (selected from pyrrolobenzodiazepines (PBD), tomaymycin, anthramycin, indolinobenzodiazepines, imidazobenzothiadiazepines, or oxazolidinobenzodiazepine dimers), calicheamicin and enediyne antibiotics, actinomycin, azaserine, bleomycin, epirubicin, tamoxifen, idarubicin, dolastatins, auristatins (monomethylauristatin E, MMAE, MMAF, auristatin PYE, 2. The conjugate compound of claim 1, wherein the conjugate compound is selected from auristatin TP, auristatin 2-AQ, 6-AQ, EB (AEB), and EFP (AEFP). ), duocarmycin, thiotepa, vincristine, hemiasterin, nazumamides, microginins, radiosumins, alterobactins, microsclerodermicins, theonellamides, esperamicins, and PNU-159682.

12. 2. The conjugated compound of claim 1 having a structure represented by the following formula 232, 308, 327, or 339: 【Chemical 54】 During the ceremony, n has the same definition as in claim 1; mAb is an antibody or cell binding molecule.

13. 13. A pharmaceutical composition for the treatment or prevention of cancer, an autoimmune disease, or an infectious disease, comprising a therapeutically effective amount of the conjugate compound of claim 1 and a pharma- ceutically acceptable salt, carrier, diluent, or excipient, or a combination thereof.

14. Said R 1 , L １ , L １’ , L1″, L ２ , L ２’ , and L ２’’ and are independently cleavable by a protease.

15. 11. A pharmaceutical composition comprising a therapeutically effective amount of the conjugate of claim 1 administered together with a synergistic agent, a chemotherapeutic agent, a radiotherapy, an immunotherapeutic agent, an autoimmune disease agent, an anti-infective agent, or another conjugate for the synergistically effective treatment or prevention of cancer, an autoimmune disease, or an infectious disease.

16. The pharmaceutical composition of claim 15, wherein the synergist is selected from one or more of the following drugs: abatacept (Orencia), abiraterone acetate (Zytiga®), acetaminophen / hydrocodone, aducanumab, adalimumab, ADXS31-142 (Advaxis'), ADXS-HER2 (Advaxis'), afatinib dimaleate (Gilotrif®), alemtuzumab (Campath®), alitretionin (Panretin®), ado-trastuzumab em (Kadcyla®), amphetamine mixed salts (amphetamine / dextroamphetamine, or adderall®), XR), anastrozole (Arimidex®), aripiprazole, atazanavir, atezolizumab (MPDL3280A), atorvastatin, axitinib (Inlyta®), avelumab, belinostat (Beleodaq™), bevacizumab (Avastin®), cabazitaxel (Jevtana®), cabozatinib (Cometriq™), bexarotene (Targretin®), blinatumomab (Blincyto™), bortezomib (Velcade®), bosutinib (Bosulif®), brentuximab Vedotin (Adcetris®), budesonide, budesonide / formoterol, buprenorphine, capecitabine, carfilzomib (Kyprolis®), celecoxib, ceritinib (LDK378 / Zykadia), cetuximab (Erbitux®), cyclosporine, cinacalcet, crizotinib (Xalkori®), Cosentyx, CTL019, dabigatran, dabrafenib (Tafinlar®), daratumab (Darzalex), darbepoetin alfa, darunavir, imatinib mesylate (Gleevec®), dasatinib (Sprycel®), denileukinDiftitox (Ontak®), denosumab (Xgeva®), depakote, dexlansoprazole, dexmethylphenidate, dexamethasone, Dignitana DigniCap Cooling System, dinutuximab (Unituxin™), doxycycline, duloxetine, duvelisib, elotuzumab, emtricitabine / rilpivirine / tenofovir disoproxil fumarate, emtricitabine / tenofovir / efavirenz, enoxaparin, enzalutamide (Xtandi®), epoetin alfa, erlotinib (Tarceva®), esomeprazole, eszopiclone, etanercept, everolimus (Afinitor®), exemceme Stan (Aromasin®), everolimus (Afinitor®), ezetimibe, ezetimibe / simvastatin, fenofibrate, filgrastim, fingolimod, fluticasone propionate, fluticasone / salmeterol, fulvestrant (Faslodex®), Gazyva, gefitinib (Iressa®), glatiramer, goserelin acetate (Zoladex), icotinib, imatinib (Gleevec), ibritumomab Tiuxetan (Zevalin®), ibrutinib (Imbruvica™), idelalisib (Zydelig®), infliximab, iniparib, insulin aspart, insulin detemir, insulin glargine, insulin lispro, interferon beta-1a, interferon beta-1b, lapatinib (Tykerb®), ipilimumab (Yervoy®), ipratropium bromide / salbutamol, ixazomi (Ninlaro®), Kanuma, lanreotide acetate (Somatuline® depot), lenalidomide, lenalidomide (Revlimid®) (Lion®), lenvatinib mesylate (Lenvima®), letrozole (Femara®), levothyroxine, lidocaine, linezolid, liraglutide, lisdexamfetamine, LN-144 (LionBiotech), MEDI4736 (AstraZeneca, Celgene), memantine, methylphenidate, metoprolol, mekinist, modafinil, mometasone, nilotinib (Tasigna®), niraparib, nivolumab (Opdivo®), ofatumumab (Arzerra®), obinutuzumab (Gazyva™), olaparib (Lynparza™), olmesartan, olmesartan / hydrochlorothiazide, omalizumab, omega-3 fatty acid ethyl esters, oseltamivir, oxycodone, palbociclib (Ibranc e®), palivizumab, panitumumab (Vectibix®), panobinostat (Farydak®), pazopanib (Votrient®), pembrolizumab (Keytruda®), pemetrexed (Alimta), pertuzumab (Perjeta™), pneumococcal conjugate vaccine, pomalidomide (Pomalyst®), pregabalin, ProscaVax, propranolol, quetiapine, rabeprazole, radium-223 chloride (Xofigo (R)), raloxifene, raltegravir, ramucirumab (Cyramza®), ranibizumab, regorafenib (Stivarga®), rituximab (Rituxan®), rivaroxaban, romidepsin (Istodax®), rosuvastatin, ruxolitinib phosphate (Jakafi™), salbutamol, sevelamer, sildenafil, siltuximab (Sylvant™), sitagliptin, sitagliptin / metformin, solifenacin, solanezumab, sorafenib (Nexavar®), sunitinib (Sutent®), tadalafil, tamoxifen, Tafinlar, talazoparib, telaprevir, temsirolimus (Toris el®), tenofovir / emtricitabine, testosterone gel, thalidomide (Immunoprin, Talidex), tiotropium bromide, toremifene (Fareston®), trametinib (Mekinist®), trastuzumab, tretinoin (Vesanoid®), ustekinumab, valsartan, veliparib, vandetanib (Caprelsa®), vemurafenib (Zelboraf®), venetoclax, vorinostat (Zolinza®), divaflibercept (Zaltrap®), and zostavax, as well as analogs, derivatives, pharma- ceutically acceptable salts thereof, carriers, diluents or excipients therefor, or combinations thereof.

Images