Sulfomaleimide-based linkers and corresponding complexes

Abstract

To provide a sulfomaleimide-based linker useful in the preparation of conjugates such as antibody-drug conjugates.SOLUTION: The present invention provides a linker of formula (I) below or a salt thereof. (X1, X2: H, a halogen or the like. L1: L1'(COZ')z' (L1': arylene, heteroarylene or the like. Z': NR4(CH2)uNR5 or the like). W: an amino acid unit. Y: para-aminobenzyl oxy-COZz (Z: NR4(CH2)uNR5 or the like). R4, R5: H or the like. c: 0, 1. q: 0-2. u: 1-6. w: 0-5. y, z, z': 0, 1. X3: H, OH or the like).SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to sulfomaleimide-based linkers useful in the preparation of conjugates such as antibody-drug conjugates (ADCs) by covalently linking a drug molecule to a binding unit, preferably an antibody.

Background Art

[0002] All ADCs are a controlled mixture of various drug-loaded species (drug molecules per antibody = DAR is 0-8), and a typical average DAR is 3.5 or 4. Unconjugated species generally have no activity and compete with drug-loaded species for binding to the antigen. In addition, species having a DAR greater than 4 have been shown to lead to lower tolerance, higher plasma clearance rates, and reduced efficacy. Most currently marketed and in-clinical-trial ADCs have common structural features such as thiosuccinimide bonds formed by the reaction of thiol and alkyl maleimide. This type of chemical reaction is widely used because the reaction of maleimide and thiol is extremely rapid and (without using either starting material species in large excess) quantitative under physiological conditions. However, thiosuccinimide formation is slowly reversible under physiological conditions. ADCs containing alkyl maleimide may result in drug loss during long-term circulation. The pharmacological consequences of the excretion of maleimide from this ADC (by retro-Michael reaction) include a decrease in the exposure of the antibody-bound drug resulting from non-target release of the drug and the linker, and a decrease in anti-tumor activity due to higher toxicity. This is the case for the thioether linker SMCC (succinimidyl 4-(N-maleimidomethyl) cyclohe ​​​​​​​​​​​​​​​Cis - cysteine - conjugated ADC and lysine - conjugated ADC via xan - 1 - carboxylate are both described in

[0003] Thus, the present invention relates to compounds useful for the conjugation of a drug and a conjugation unit, and the resulting conjugate is ​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​n -Ariren-(CH2) p -,-(CH2) n -Heterorialene-(CH2) p -,-(CH2) n -Cycloalkanediyl-(CH2) p -,-(CH2CH2O) m -CH2-CH2-Arirene-(CH2) p -,-(CH2CH2O) m -CH2- CH2-heteroarylene-(CH2) p -,-(CH2CH2O) m -CH2-CH2 -Cycloalkanediyl-(CH2) p -,-(CH2) n -Ariren-CH2-CH 2-(OCH2CH2) m -,-(CH2) n -heterorialene-CH2-CH2-( OCH2CH2) m -, or -(CH2) n -Cycloalkanedil-CH2-CH2 -(OCH2CH2) m -and; Each W independently represents an amino acid unit; Y is -PAB-CO-(Z) z -And here, PAB is, [ka] (The oxygen in the PAB unit is CO-(Z) z (and is connected to); Z is -NR4-(CH2) u -NR5- or -NR4-(CH2) u -NR5- CO-, or even -NR4-(CH2) u -NR5-CO-(CH2) v -too or -NR4-(CH2) u -NR5-CO-(CH2)v -CO- (where the NR 4 groups are linked to the CO group of PAB-CO); Z’ is -NR4-(CH2) u -NR5- or -NR4-(CH2) u -NR5- CO-(CH2) v - (where the NR4 group is linked to the CO group of CO-Z’) ; R4 and R5 are independently H or a (C1-C6) alkyl group; c is 0 or 1, preferably 1; m is an integer from 1 to 15; n is an integer from 1 to 6; p is an integer from 1 to 6; q is 0, 1 or 2, preferably 2; r is an integer from 1 to 24, particularly from 1 to 12; u is an integer from 1 to 6; v is an integer from 1 to 6; w is an integer from 0 to 5, preferably 0 or 2; y is 0 or 1 (preferably, when w is 0, y is 0, and when w is an integer from 1 to 5 y is 0 or 1); z is 0 or 1; z’ is 0 or 1, particularly 0; and X3 represents H when y = z = 1 and Z is -NR4-(CH2) u -NR5-, or when c = w = y = 0, z’ = 1 and Z’ is -NR4-(CH2) u -NR5-, and represents OH, NH2 or a leaving group in other cases. The leaving group is, more specifically, a halogen atom, a sulfonate of the formula -OSO2-R

[0005] a succinimidyl oxy, 4-nitro-phenoxy, pentafluorophenoxy or LG ; N-succinimidyl oxy, 4-nitro-phenoxy, pentafluorophenoxy or It is N-benzotriazoloxy, and R LG (C1-C6) alkyl, aryl, and Reel-(C1-C6)alkyl or (C1-C6)alkyl-aryl group, preceding The base may optionally be substituted with one or more halogen atoms, such as fluorine atoms. .

[0006] Preferably, the compound of formula (I) is X1 is Cl, X2 is H, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is Cl; X1 is Cl, X2 is Cl, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is Cl; X1 is Cl, X2 is H, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is Cl; X1 is Cl, X2 is H, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is Cl; X1 is Cl, X2 is H, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is Cl; X1 is Cl, X2 is H, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is Br; X1 is Cl, X2 is H, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is I; X1 is H, X2 is Cl, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is Cl; X1 is H, X2 is Br, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is Cl; X1 is H, X2 is Br, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is Cl; X1 is Cl, X2 is Cl, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is Cl; X1 is Cl, X2 is Cl, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is Cl; X1 is Cl, X2 is Cl, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is Cl; X1 is Cl, X2 is Br, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is Cl; X1 is Cl, X2 is Br, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is Cl; X1 is Cl, X2 is Br, q is 0, and L1 is [ka] And c is 0, w is 0, y is 0, and X3 is Cl; or X1 is Br, X2 is Br, q is 0, and L1 is [ka] In a compound where c is 0, w is 0, y is 0, and X3 is Cl, without; Here, the dashed line is L1 and [ka] The bond points with the nitrogen atom are shown, and the dashed lines indicate the bond points between L1 and X3.

[0007] These compounds are disclosed in WO2007 / 001932 or US4,127,687. However, more specifically, it is intended as a linker for the fabrication of complexes such as ADCs. There isn't one.

[0008] The present invention also relates to the following formula (II): [ka] Preferably, the following formula (IIa): [ka] With respect to the linker drug complex or a salt thereof, During the ceremony, X1 and X2 are independent of each other: H, halogen atom, (C1-C6) alkoxy, field Aryloxy or -O-(CH2CH2O) which may be substituted by compounding r H X1 and X2 cannot represent H simultaneously; L1 is given by the formula L1'-(CO-Z') z’ This represents the base, where L1' is -(CH2) n -,-(CH2CH2O) m -CH2-CH2-, Arylene, Heteroarylene, Si Chloalkanediyl, -(CH2) n -Ariren-, -(CH2) n - Heteralile Hmm, -(CH2) n -Cycloalkanediyl-, -Arirene-(CH2) p -, -he Terrorarylene-(CH2) p -, -cycloalkanediyl-(CH2) p -,-(CH 2) n -Ariren-(CH2) p -,-(CH2) n -Heterorialene-(CH2) p -,-(CH2) n -Cycloalkanediyl-(CH2) p-,-(CH2CH2O) m -CH2-CH2-Arirene-(CH2) p -,-(CH2CH2O) m -CH2- CH2-heteroarylene-(CH2) p -,-(CH2CH2O) m -CH2-CH2 -Cycloalkanediyl-(CH2) p -,-(CH2) n -Ariren-CH2-CH 2-(OCH2CH2) m -,-(CH2) n -heterorialene-CH2-CH2-( OCH2CH2) m -, or -(CH2) n -Cycloalkanedil-CH2-CH2 -(OCH2CH2) m -and; Each W independently represents an amino acid unit; Y is -PAB-CO-(Z) z -And here, PAB is, [ka] (The oxygen in the PAB unit is CO-(Z) z (and is connected to); Z is -NR4-(CH2) u -NR5- or -NR4-(CH2) u -NR5- CO-, or even -NR4-(CH2) u -NR5-CO-(CH2) v -too or -NR4-(CH2) u -NR5-CO-(CH2) v -CO- (the above NR Four units are connected to the CO group of PAB-CO. Z' is -NR4-(CH2) u -NR5- or -NR4-(CH2) u -NR5- CO-(CH2) v - (The NR4 group is linked to the CO group of CO-Z') ; R4 and R5 are independently H or (C1-C6) alkyl groups; Q represents the drug portion; c is 0 or 1, preferably 1; m is an integer between 1 and 15; n is an integer between 1 and 6; p is an integer between 1 and 6; q is 0, 1, or 2, preferably 2; r is an integer between 1 and 24, in particular between 1 and 12; u is an integer between 1 and 6; v is an integer between 1 and 6; w is an integer between 0 and 5, preferably 0 or 2; y is 0 or 1 (preferably, if w is 0, y is 0 and w is 1 to 5) If y is an integer, then y is either 0 or 1. z is either 0 or 1; and z' is either 0 or 1, in particular, 0.

[0009] The present invention also relates to the following formula (III) or (IV): [ka] With regard to the binding unit drug complex or a salt thereof, preferably a pharmaceutically acceptable salt thereof, During the ceremony, The binding units are peptides, proteins (e.g., manipulated proteins), antibodies (e.g., mono- Clonal antibodies) or their antigen-binding fragments; L1 is given by the formula L1'-(CO-Z') z’ This represents the base, where L1' is -(CH2) n -,-(CH2CH2O) m -CH2-CH2-, Arylene, Heteroarylene, Si Chloalkanediyl, -(CH2)n -arylene-, -(CH2) n -heteroarylene -, -(CH2) n -cycloalkanediyl-, -arylene-(CH2) p -, -he teroarylene-(CH2) p -, -cycloalkanediyl-(CH2) p -, -(CH 2) n -arylene-(CH2) p -, -(CH2) n -heteroarylene-(CH2) p -, -(CH2) n -cycloalkanediyl-(CH2) p -, -(CH2CH2O) m -CH2-CH2-arylene-(CH2) p -, -(CH2CH2O) m -CH2- CH2-heteroarylene-(CH2) p -, -(CH2CH2O) m -CH2-CH2 -cycloalkanediyl-(CH2) p -, -(CH2) n -arylene-CH2-CH 2-(OCH2CH2) m -, -(CH2) n -heteroarylene-CH2-CH2-( OCH2CH2) m -, or -(CH2) n -cycloalkanediyl-CH2-CH2 -(OCH2CH2) m -; Each W independently represents an amino acid unit; Y is -PAB-CO-(Z) z -, where PAB is

Chemical Structure

[0010] According to a preferred embodiment, the binding unit is an IGF-1R antibody, a HER2 antibody (for example, Trastuzumab) or its antigen-binding fragment.

[0011] The present invention also relates to a drug to an antibody (e.g., a monoclonal antibody) or its antigen-binding flag. A linker of formula (I) or formula (II) for covalently linking to bonding units such as ments. This relates to a drug linker complex (preferably q=2) of the phosphate group. Such a covalent bond is phosphorus It is produced by the car part.

[0012] In fact, preferably, the compound of formula (I) or (II) with q=2 is the drug that acts as an antibody (for example) For example, a covalent bond to a binding unit such as a monoclonal antibody or its antigen-binding fragment. It is useful for linking to.

[0013] Compounds of formula (I) or (II) with q=0 or 1 are also compounds of formula (I) with q°=°2. It can also be used as a synthetic intermediate to prepare compound (II). In conclusion, Furthermore, the present invention also relates to a synthetic intermediate of the formula (I) defined above, where q = 0 or 1. ) or (II) relating to compounds.

[0014] The present invention also relates to a linker of formula (I) or formula (II), (III) or (IV) This relates to a method for creating a complex.

[0015] The present invention also relates to a binding unit drug conjugate of formula (III) or (IV) and at least one This relates to a pharmaceutical composition comprising a pharmaceutically acceptable excipient.

[0016] The present invention also relates to the conclusion of formula (III) or (IV) for use in the treatment of cancer. Combined drug complexes, or formula (III) or (IV) bound unit drug complexes, and at least The present invention relates to a pharmaceutical composition comprising one pharmaceutically acceptable excipient.

[0017] The present invention also relates to the manufacture of a drug intended for use in the treatment of cancer, formula ( Regarding the use of conjugated unit drug conjugates as described in (III) or (IV).

[0018] The present invention also provides a bonding unit of formula (III) or (IV) in an effective amount to the target object as needed. Drug conjugate or binding unit drug conjugate of formula (III) or (IV) and at least one other A treatment for cancer comprising administering a pharmaceutical composition comprising a pharmaceutically acceptable excipient of the same type. Regarding methods for doing so.

[0019] definition In this invention, the term "pharmaceutically acceptable" means useful in the preparation of a pharmaceutical composition. And for pharmaceutical use, it is intended to mean something that is generally safe and non-toxic.

[0020] The term "pharmaceutically acceptable salt" in the context of this invention means a drug as defined above. This means a salt of a compound that is scientifically acceptable and has the pharmacological activity of the corresponding compound. It is intended.

[0021] Pharmaceutically acceptable salts include: (1) Acids formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Addition salts; or acetic acid, benzenesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, Glutamic acid, glycolic acid, hydroxynaphthoic acid, 2-hydroxyethanesulfonic acid Lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, muconic acid, 2-naphtha Lensulfonic acid, propionic acid, succinic acid, dibenzoyl-L-tartaric acid, tartaric acid, p- Formed with organic acids such as leuen sulfonic acid, trimethylacetic acid, and trifluoroacetic acid. The acid addition salts that are produced, and (2) The acid proton of the compound is an alkali metal ion, an alkaline earth metal ion, or Substituted with metal ions such as aluminum ions; or organic or non-organic. Salts formed when an autosomal base is coordinated. This includes. Acceptable organic bases include diethanolamine, ethanolamine, and N-methylamine. Contains tilglucamine, triethanolamine, tromethamine, etc. Acceptable inorganic The bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, and sodium carbonate. It also contains sodium hydroxide.

[0022] The term "halogen" as used herein refers to fluorine, bromine, chlorine, or iodine. It refers to elementary atoms.

[0023] The term "(C1-C6) alkyl" as used herein refers to a group of 1 to 6 carbon atoms. It refers to, but is not limited to, monovalent, linear or branched saturated hydrocarbon chains containing atoms. Methyl, ethyl, n-propyl, iso-propyl, n-butyl, isobutyl, sec- Contains butyl, t-butyl, n-pentyl, n-hexyl, etc.

[0024] The term "(C1-C6) alkoxy" as used herein refers to a compound involving an oxygen atom. This refers to an alkyl group (C1-C6) as defined above, which is bonded to a molecule, and is limited to It is not done, but methoxy, ethoxy, n-propoxy, iso-propoxy, n- Butoxy, iso-butoxy, sec-butoxy, t-butoxy, n-pentoxy, n-hex Includes xoxy, etc.

[0025] The term "(C2-C6) alkenyl" as used herein refers to 2 to 6 carbon atoms. A linear or branched monovalent unsaturated molecule containing elementary atoms and at least one double bond. It refers to and is not limited to a hydrocarbon chain, but includes ethenyl, propenyl, butenyl, and pe Includes anthenyl, hexenyl, etc.

[0026] The term "cycloalkanediyl," as used herein, is advantageously defined as 4 to 10 This refers to, and is not limited to, a divalent hydrocarbon ring having 5 or 6 carbon atoms. It is not a good choice, but it contains cyclopentanediyl, cyclohexanediyl, etc. That is a cyclohexanediyl group.

[0027] The term "aryl," as used herein, refers to, for example, a phenyl group or naphth. Preferably, it comprises 6 to 10 carbon atoms, such as a chill group, and has one or more fused rings. This refers to the monovalent aromatic hydrocarbon group contained within. Advantageously, this would be a phenyl group.

[0028] The term "aryl-(C1-C6)alkyl" as used herein means the above Substituted with an aryl group as defined above, (C1-C6) aryl group as defined above This refers to a lukyl group. In particular, it could be a benzyl group.

[0029] The term "(C1-C6) alkyl-aryl" as used herein means the above Substituted with (C1-C6) alkyl groups as defined above, This refers to a reel group. In particular, it could be a toryl group (CH3Ph).

[0030] The term "aryloxy," as used herein, refers to a molecule via an oxygen atom. It refers to, but is not limited to, a bonded aryl group as defined above, but Contains phenyloxy.

[0031] The term "arirene," as used herein, refers, for example, to a phenylene group or Preferably, it contains 6 to 10 carbon atoms, such as a naphthylene group, and has 1 or more condensation This refers to a divalent aromatic hydrocarbon group containing a ring. Advantageously, it would be a phenylene group. .

[0032] The term "heteroarylene," as used herein, refers to one or more, in particular , 1 to 4, preferably 1 to 3 carbon atoms, each consisting of a sulfur atom, an oxygen atom, and a nitrogen atom. A heteroatom selected from elementary atoms, preferably selected from oxygen and nitrogen atoms. , more preferably, one or more, in particular one or two, which are substituted with nitrogen atoms. This refers to a divalent aromatic group containing a condensed hydrocarbon ring. Advantageously, it is a divalent 1,2,3- Reazoles, for example, divalent 1H-1,2,3-triazoles.

[0033] When used in this specification, the term "leaving group" refers to a nucleophile in a nucleophilic substitution reaction. (For example, amines or alcohols having functional groups NH or OH respectively) This refers to chemical groups that can be replaced. Such leaving groups include, in particular, halogen atoms, sulfonates, and N -Succinimidyloxy, 4-nitrophenyloxy, pentafluorophenoxy It may be N-benzotriazoloxy. Sulfonates in particular have the group -OSO2-R LG And here, R LG (C1-C6) alkyl, aryl, aryl-(C1-C6 ) represents an alkyl or (C1-C6)alkyl-aryl group, wherein the group is one or more They may be substituted with halogen atoms such as fluorine atoms. Sulfonates are particularly In addition, mesylate (OMs, CH3-S(O2)O-), triflate (OTf, CF3- S(O)2O- or tosylates (OTs, p-Me-C6H4-S(O)2O-) It is possible. The leaving groups are, in particular, Cl, Br, I, OTf, OMs, OTf, and N-sucrose. It may be midyloxy, 4-nitrophenyloxy, or N-benzotriazoloxy. .

[0034] The term "trialkylsilyl group" is used herein as -SiAlk1A The term lk2Alk3 refers to the Alk1, Alk2, and Alk3 group, and Alk1, Alk2, and Alk3 are the same or different. It represents a (C1-C6)-alkyl group as defined below. For example, it is trimethylalkyl It may be a lyl group or a triethylsilyl group.

[0035] The term "protective form" of a molecule refers to the presence of at least an OH or NH functionality in the molecule. This means that each group is protected by either an O-protecting group or an N-protecting group.

[0036] The term "protecting group," as used herein, refers to a certain reactive part of a polyfunctional compound. The site is selectively blocked so that chemical reactions at other unprotected reactive sites can selectively proceed. It refers to a chemical group.

[0037] The term "O-protecting group" is used herein in reference to "Greene's Protective Gro "ups In Organic Synthesis", 4th edition, 2007, John Wiley & Sons, Hoboken, New Undesirable reactions during the synthesis procedure, such as O-protecting groups disclosed in Jersey, can occur. This refers to substituents that protect the hydroxyl group (OH). Hydroxyl group protected by an O-protecting group. The group can be, for example, an ether, ester, carbonate, or acetal. The O-protecting group has one or more (especially 1 to 3) halogen atoms (e.g., chlorine atoms). (C1-C6) alkyl groups may be substituted in some cases, for example, methyl, ethyl, tert-butyl or 2,2,2-trichloroethyl; aryl-(C1-C6) aryl Kill, for example, benzyl (the aryl portion may be arranged with one or more methoxy groups) (may be substituted), for example, benzyl (Bn) or p-methoxybenzyl (PMB) ); Trityl derivatives of formula -CAr1Ar2Ar3, for example, triphenylmethyl(trityl) (Also called ru-Tr), (4-methoxyphenyl)diphenylmethyl(methoxytrityl- Also called NMT) or bis-(4-methoxyphenyl)phenylmethyl (dimethoxymethyl) Lithyl-DMT (also known as formula CH2OR) GP2 or -CH2SR GP2 (especially,- CH2OR GP2 ) Substituted methyl groups, for example, methoxymethyl (MOM), benzyl hydroxymethyl methyl, 2-methoxyethoxymethyl (MEM), 2-(trimethylsilyl)ethoxy Methyl or methylthiomethyl; formula -CH2CH2OR GP2 or -CH2CH2SR GP2 (Especially -CH2CH2OR GP2 ) Substituted ethyl group, for example, ethoxyethyl ( EE); Formula -SiR GP3 R GP4 R GP5 The silyl group, for example, trimethylsilyl (T MS), triethylsilyl (TES), t-butyldimethylsilyl (TBS or TBD) MS) and t-butyldiphenylsilyl (TBDPS); formula -CO-R GP6 Carbonara Nylating groups, for example, acetyl (Ac), pivaloyl (Piv or Pv), or ben Zoyl (Bz) or formula -CO2-R GP7 The silyl group, for example, allyloxycarbon Alloc or 9-Fluorenylmethyloxycarbonyl (Fmoc); or tetrahydropyranyl ( [ka] ) (THP) or tetrahydrofuranil ( [ka] ) may be a base; where Ar1, Ar2 and Ar3 are independent of each other, one or more R represents an aryl group such as phenyl, which may be optionally substituted with a methoxy group; GP2 These include aryl (e.g., phenyl), (C1-C6) alkoxy (e.g., methoxy), and Alternatively, it may be optionally substituted with a trialkylsilyl group (e.g., SiMe3). (C1-C6) alkyl (e.g., methyl or ethyl); R GP3 , R GP4 Oh biR GP5 These are (C1-C6) alkyl or aryl (e.g., phenyl R) represents a group; and, GP6 and R GP7 These are, independently of each other, (C1-C6) Alky (C2-C6) alkenyl, aryl, aryl-(C1-C6) alkyl or 9 - Represents a fluorenylmethyl group.

[0038] The term "N-protecting group," as used herein, refers to an undesirable reaction during the synthesis procedure. This refers to a group intended to protect an amine functional group (especially a primary amine functional group) from an external source. Commonly used N-protecting groups are described in “Greene’s Protective Groups In Organic Synthesis”. Disclosed in the 4th edition, 2007, John Wiley & Sons, Hoboken, New Jersey. The amine functional groups protected by the N-protecting group are carbamates, amides, and sulfonamides. N-alkyl derivatives, aminoacetal derivatives, N-benzyl derivatives, imine derivatives, It may be an enamine derivative or an N-heteroatom derivative. In particular, the N-protecting group is formyl aryl groups such as phenyl, which may optionally be substituted with one or more methoxy groups. For example, p-methoxyphenyl (PMP); the aryl moiety consists of one or more methoxy groups. aryl-(C1-C6)alkyl groups such as benzyl, which may be substituted in some cases. For example, benzyl (Bn), p-methoxybenzyl (PMB), or 3,4-dimethoxybenzyl Benzyl (DMPM);-CO-R GP1 For example, acetyl (Ac), pivaloyl (P iv or Pv), benzoyl (Bz) or p-methoxybenzylcarbonyl (Mo z);-CO2-R GP1 For example, t-butyloxycarbonyl (Boc), trichloro Ethoxycarbonyl (TROC), allyloxycarbonyl (Alloc), benzyl Xycarbonyl (Cbz or Z) or 9-Fluorenylmethyloxycarbonyl ( Fmoc);-SO2-R GP1 For example, phenylsulfonyl, tosyl (Ts or Tos) or 2-nitrobenzenesulfonyl (also called nosyl-Nos or Ns) etc; Here, R GP1It may be substituted with one or more halogen atoms such as F or Cl. It may represent an (C1-C6) alkyl group; a (C2-C6) alkenyl group, for example, Allyl; one or more groups selected from OMe (methoxy) and NO2 (nitro) Aryls such as phenyl, which may be substituted depending on the circumstances; one or more aryl moieties. The aryl-(C1-C) group, such as benzyl, may be optionally substituted with a methoxy group. 6) Alkyl; or 9-fluorenylmethyl group.

[0039] The terms "antibody," "ab," "Ab," "MAb," or "immunoglobulin" are used in a broad sense. These are used interchangeably and include monoclonal antibodies, isolated, manipulated or recombinant antibodies. Body (e.g., full-length or complete monoclonal antibody), polyclonal antibody, multivalent antibody, This includes multispecific antibodies (e.g., bispecific antibodies) and also insofar as they exhibit the desired biological activity. This also includes antibody fragments from [the researchers].

[0040] The term "recombinant antibody" refers to an antibody produced from the expression of recombinant DNA within living cells. The recombinant antibodies developed by Akira produce DNA sequences not found in biological organisms, as described by those skilled in the art. This is obtained by using laboratory methods for genetic engineering of knowledge.

[0041] The term "antigen-binding fragment" of an antibody according to the present invention refers to the target of the antibody (generally, an anti- Any peptide, polypeptide, or protein that retains the ability to bind to (the origin). The intention is to demonstrate this.

[0042] "Binding" and "binding" refer to the process by which an antibody, or its antigen-binding fragment, interacts with an antigen. The intention is to form a complex that is relatively stable under physical conditions. Specific binding is minimal. Even without it, it's approximately 1 x 10 -6 It can be characterized by the equilibrium dissociation constant of M. Whether the two molecules bind or not. Methods for determining this are well known in the art, for example, equilibrium dialysis, surface plasmo This includes resonance, radiolabeling assays, etc. To avoid suspicion, it is assumed that the aforementioned antibody is different This does not mean that it cannot bind to or interfere with the antigen at low levels. However, In one embodiment, the antibody binds only to the antigen.

[0043] In this specification, the expression "IGF-1R antibody" means "anti-IGF-1 This should be interpreted similarly to "R antibody," meaning an antibody capable of binding to IGF-1R.

[0044] In this specification, the term "HER2 antibody" is used interchangeably with "anti-HER2 antibody." It should be interpreted similarly, meaning an antibody capable of binding to HER2.

[0045] Half-effective concentration (EC) 50 The term refers to the baseline and the maximum after a certain exposure time. This corresponds to the concentration of a drug, antibody, or toxin that induces half of the response between the maximum values. It is commonly used as a measure of potency. Therefore, the EC of the dose-response curve 50 Its maximum effect This represents the concentration of the compound found in 50% of the fruit. (Quantal dose response curve) EC (curve) 50 This refers to the concentration of a compound that causes 50% of the population to react after a specific exposure period. To represent. Concentration measurements generally show a sigmoid curve that increases rapidly with relatively small changes in concentration. This can be mathematically determined by deriving the best-fitting line.

[0046] In a preferred embodiment, the EC determined in the present invention is 50 It is exposed on human tumor cells. This characterizes the efficacy of the antibody that binds to IGF-1R ECD. 50 Parame The ta is determined using FACS analysis. EC 50 The parameters are on human tumor cells. This represents the antibody concentration that yields 50% of the maximum binding to expressed human IGF-1R. 50 The values ​​are from a 4-parameter regression curve fitting program (Prism software). It is calculated as the midpoint of the dose-response curve using ). This parameter is physiological / pathological. These were selected to be typical of physical conditions.

[0047] The term "epitope" refers to the region of an antigen to which an antibody binds. Epitopes are structural and Functional epitopes can be defined as functional epitopes. Functional epitopes are generally a subset of structural epitopes. It has residues that directly contribute to the affinity of the interaction. Epitopes are also conformational In some cases, this may be composed of nonlinear amino acids. In certain embodiments, Pitopes are chemically active, such as amino acids, sugar side chains, phosphoryl groups, or sulfonyl groups. It may include a group of surface molecules that are determinants, and in certain embodiments, certain three-dimensional structural features, and / or may have specific charge characteristics.

[0048] The terms "monoclonal antibody" or "Mab" are used herein in a manner that is not applicable to the actual application of the term. This refers to antibodies obtained from a qualitatively homogeneous population of antibodies; that is, the individual antibodies in that population are, They are identical except for the possibility of trace amounts of latent natural mutations. Monoclonal antibodies have specificity. It has high fidelity and is directed towards a single epitope. Such monoclonal antibodies target a single B cell. Monoclonal antibodies can be produced by a single clone or hybridoma. It may be a substitute type, that is, it can be produced by protein engineering or chemical synthesis. Monoclonal antibodies may also be isolated from phage antibody libraries. In addition, Generally, polyclonal antibodies containing various antibodies against various determinants or epitopes are produced. In contrast to monoclonal antibodies, each monoclonal antibody is directed to a single epitope of the antigen. In this document, monoclonal antibodies include mouse, chimeric, and humanized antibodies.

[0049] The term "chimeric antibody" refers to an antibody derived from an antibody of a given species, which has a naturally variable region (light chain and heavy chain) derived from that region. , an antibody containing the constant regions of the light chain and heavy chain of an antibody of a different species from the given species mentioned above. Regarding this, chimeric antibodies can be produced using recombinant genetics techniques. For example, chimeric antibodies are promoters, and non-human, particularly mouse monoclonal antibodies. Sequences encoding the variable region of the body, and heterologous, preferably human antibody constant region This can be produced by cloning recombinant DNA containing a sequence. The chimeric antibody according to the present invention, encoded by a single recombinant gene, is, for example, used in mice. -It may be a human chimera, and the specificity of this antibody is determined by a variable region derived from mouse DNA. Its isotype is determined by a constant region derived from human DNA.

[0050] The term "humanized antibody" refers to an antibody that contains a CDR region derived from a non-human antibody, and also includes other components of the antibody molecule. The part refers to antibodies derived from one (or more) human antibodies. In addition, the skeletal segment Some of the nucleotide residues (referred to as FRs) can be modified to preserve binding affinity. Humanized antibodies or their fragments can be produced by techniques known to those skilled in the art. Such humanized antibodies can be used for in vitro or in vivo diagnosis. They are preferable for use in methods that include preventive and / or therapeutic measures. For example , patents and patent applications EP0451216, EP0682040, EP09 by PDL 39127, EP0566647, US5,530,101, US6,180,370, The term "CDR graphics" is described in US5,585,089 and US5,693,761. Other humanization techniques, such as "feeting" technology, are also known to those skilled in the art. (U.S. Patent No. 5,639) Nos. 641, 6,054,297, 5,886,152 and 5,877 Issue 293 can also be cited.

[0051] Unless otherwise specified herein, the complementarity determination region or CDR is defined as IMGT. The hypervariability of the heavy and light chains of immunoglobulins, as defined according to the immunoglobulin system. It means "area" or "domain."

[0052] However, CDR also uses the Kabat numbering system (Kabat et al., Seque nces of proteins of immunological interest, 5th edition, US Department of Health and It can also be defined according to Human Services, NIH, 1991 and subsequent editions. There are heavy chain CDRs and three light chain CDRs. Here, the term "CDR" is used in some cases. And the majority of the amino acids are responsible for the binding affinity of antibodies to the antigen or epitope that it recognizes. This is used to indicate one or more, or even all, regions containing no acid residues. Interpretation of this application To simplify things, we will not define CDR by Kabat. However, IMGT It is obvious to those skilled in the art that using the definition of CDR by [the relevant source] to define CDR by Kabat is [the relevant source]. be.

[0053] In the sense of the present invention, "identity" or "sameness" between two nucleic acid sequences or amino acid sequences The "uniformity percentage" is obtained after optimal alignment. This refers to the percentage of a single nucleotide or amino acid residue, and this percentage is It is purely statistical, and the difference between the two sequences is randomly distributed along their lengths. There is. The comparison of two nucleic acid sequences or amino acid sequences is conventionally done to determine which is the best ally. This is done by comparing the sequences after the comparison, and the comparison is done by segment or " This can be done by using the "Alignment Window". Optimal alignment of the array for comparison. In addition to manual comparison, Smith and Waterman (1981) [Ad. App. Math. 2:48] 2] Local homology algorithm method, Neddleman and Wunsch (1970) [J. Mol. Biol. Local homology algorithms of [48:443], Pearson and Lipman (1988) [Proc. Nat Using the similarity search methods described in [l. Acad. Sci. USA 85:2444], or their algorithms The means of computer software (Wisconsin Genetics Software Package, Genetic s Computer Group, 575 Science Dr., Madison, WI GAP, BESTFIT, FAST A and TFASTA or comparison software BLAST NR or BLAST P It can be done by ( ).

[0054] The identity percentage is determined by the amino acid nucleotides between two sequences, preferably between two complete sequences. Determine the number of identical positions of octides or residues, and use the number of identical positions as the alignment number. Divide by the total number of positions in the window, and multiply the quotient by 100 to get the percentage of identity between the two sequences. It is calculated by obtaining the value of .

[0055] For example, BLAS is available at the website http: / / www.ncbi.nlm.nih.gov / gorf / bl2.html T program “BLAST2 sequences” (Tatusova et al., “Blast 2 sequence ences - a new tool for comparing protein and nucleotide sequences”, FEMS Microb iol., 1999, Lett. 174:247-250) default parameters (especially "open gap") The parameters for "Penalty": 5 and "Extension Gap Penalty": 2 Regarding the matrix; the matrix to be selected is, for example, proposed by this program. It can be used with the "BLOSUM 62" matrix; compare the two distributions. The percentage of identity between columns is calculated directly by this program.

[0056] The term "reverse mutation" refers to a mutation in which a human residue present in the germline has been originally introduced into the mouse sequence. This refers to a mutation or substitution by a corresponding residue that already exists.

[0057] Terms used interchangeably in this specification include "nucleic acid," "nuclear sequence," "nucleic acid sequence," and "po "Dinucleotide," "oligonucleotide," "polynucleotide sequence," and "nucleate" An "Otid sequence" defines a nucleic acid fragment or region that is modified or unmodified. Meaning, containing or not containing non-natural nucleotides, double-stranded DNA, single-stranded DNA or This refers to the exact nucleotide sequence of the transcript of the aforementioned DNA.

[0058] The term "peptide" refers to amino acids linked together by peptide (amide) bonds. Regarding the chain of M. A covalent peptide bond (amide) is formed by the carboxyl group (C) of a certain amino acid. It is formed by reacting the OOH group with the amino group (NH2) of another amino acid. The term "peptide" includes oligopeptides and polypeptides.

[0059] The term "protein" is composed of post-translational modifications and other processes that make it biologically functional. A collection of one or more peptides as defined above that are undergoing protein folding. It is the body.

[0060] The term "amino acid," as used herein, refers to D-type or L-type natural α-amino acids. ano acids (for example, alanine (Ala), arginine (Arg), asparagine (Asn), Aspartic acid (Asp), cysteine ​​(Cys), glutamine (Gln), glutamine Acid (Glu), Glycine (Gly), Histidine (His), Isoleucine (Ile), Leucine (Leu), Lysine (Lys), Methionine (Met), Phenylanine (P) he), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), and valine (Val), as well as unnatural amino acids. (For example, β-alanine, allylglycine, tert-leucine, 3-amino-adipine) Acids, 2-aminobenzoic acid, 3-aminobenzoic acid, 4-aminobenzoic acid, 2-aminobutane Acid, 4-amino-1-carboxymethylpiperidine, 1-amino-1-cyclobutancal 4-aminocyclohexaneacetic acid, 1-amino-1-cyclohexanecarboxylic acid (c (1R,2R)-2-aminocyclohexanecarboxylic acid (1R,2S)-2-aminocyclohexanecarboxylic acid, (1S,2R)-2-amino Cyclohexanecarboxylic acid, (1S,2S)-2-aminocyclohexanecarboxylic acid, 3 -aminocyclohexanecarboxylic acid, 4-aminocyclohexanecarboxylic acid, (1R,2 R)-2-aminocyclopentanecarboxylic acid, ( (lR,2S)-2-aminocyclopentanecarboxylic acid, 1-amino-1-cyclopentane Carboxylic acid, 1-amino-1-cyclopropanecarboxylic acid, 4-(2-aminoethoxy) -benzoic acid, 3-aminomethylbenzoic acid, 4-aminomethylbenzoic acid, 2-aminobutane Acids, 4-aminobutanoic acid, 6-aminohexanoic acid, 1-aminoindan-1-carboxylic acid , 4-aminomethylphenylacetic acid, 4-aminophenylacetic acid, 3-amino-2-naphtho E acid, 4-aminophenylbutanoic acid, 4-amino-5-(3-indolyl)-pentanoic acid (4R,5S)-4-amino-5-methylheptanoic acid, (R)-4-amino-5-methyl (R)-4-amino-6-methylthiohexanoic acid, (S)-4-amino- Pentanoic acid, (R)-4-amino-5-phenylpentanoic acid, 4-aminophenylpropionic acid Onic acid, (R)-4-aminopimelic acid, (4R,5R)-4-amino-5-hydroxy Hexanoic acid, (R)-4-amino-5-hydroxypentanoic acid, (R)-4-amino-5-(p-hydroxyphenyl)-pentanoic acid, 8-aminoocta (2S,4R)-4-amino-pyrrolidine-2-carboxylic acid, (2S,4S)-4 -amino-pyrrolidine-2-carboxylic acid, azetidine-2-carboxylic acid, (2S,4R) -4-benzyl-pyrrolidine-2-carboxylic acid, (S)-4,8-diaminooctanoic acid, tert-butylglycinate, γ-carboxyglutamate, β-cyclohexylalanine Citrulline, 2,3-diaminopropionic acid, hippuric acid, homocyclohexylalanine Moleucine, homophenylalanine, 4-hydroxyproline, indol n-2-carboxylic acid, isonipecotinic acid, α-methylalanine, nicopetinic acid (nicopet ic acid), norleucine, norvaline, octahydroindole-2-carboxylic acid, ol Nitin, penicillamine, phenylglycine, 4-phenyl-pyrrolidine-2-carboxylic acid Pipecolic acid, propargylglycine, 3-pyridinylalanine, 4-pyridinylalanine Nin, 1-pyrrolidine-3-carboxylic acid, sarcosine, statin, tetrahydroisoquino Phosphorus-1-carboxylic acid, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, (or tranexamic acid) Specific description of the invention

[0061] Detailed explanation Linker section The linker portion according to the present invention covalently binds an antibody to at least one drug portion. It can be done.

[0062] The linker portion may be "impossible to cut" or "cuttable."

[0063] In a preferred embodiment, it is contained in a "cleavable" linker portion, and drugs are present within the cell. It promotes the release of [something].

[0064] For example, in some embodiments, the linker is located in the intracellular environment (e.g., lysosomes). It can also be cleaved by cleavage factors present in endosomes or caveolae. The incubator is, for example, but is not limited to, a lysosome or endosomal protea. Peptidyl, which is cleaved by intracellular peptidase or protease enzymes, including -ase. It can be a linker. Generally, a peptidyl linker is at least two consecutive amino acids It comprises an acid or at least three consecutive amino acids. The cleavage factor is cathepsin B and It may contain D and plasmin, all of which transform dipeptide drug derivatives within target cells. It is known that hydrolysis leads to the release of active drugs. For example, high expression in cancer tissue. Peptidyl linkers that can be cleaved by the thiol-dependent protease cathepsin-B (e.g.) For example, a linker containing Phe-Leu or Gly-Phe-Leu-Gly It is usable. In certain embodiments, peptides that can be cleaved by intracellular proteases The zill linker may consist of Val-Cit, Phe-Lys, or Val-Ala. , or it is. One advantage of using intracellular proteolytic drug release is that the drug Generally, when complexes are formed, they are attenuated, and the serum stability of these complexes is generally high. That is the case.

[0065] Base-L1-(CO) c - represents the stretcher unit of the linker section, which is always present. Base-L1-(CO) c - is the formula -L1'-(CO-Z') z’ -(CO) c - is the basis Here, z' and c are either 0 or 1, for example, if z' is 0, then base L 1'-(CO) c - is preferred if at least one of w and y is not 0, z' is 0 in all other cases, and z' is either 0 or 1 in all other cases.

[0066] L1' is -(CH2) n -,-(CH2CH2O) m -CH2-CH2-, Allire N, heteroarylene, cycloalkanediyl, -(CH2) n -Ariren-, -(C H2) n -Heterorialene-, -(CH2) n -Cycloalkanedil-, -Allire Hmm-(CH2) p -,-heterorialene-(CH2) p -, -cycloalkanediyl- (CH2) p -,-(CH2) n -Ariren-(CH2) p -,-(CH2) n -Hete Roariren-(CH2) p -,-(CH2) n -Cycloalkanediyl-(CH2) p -,-(CH2CH2O) m -CH2-CH2-Arirene-(CH2) p -,-(CH 2CH2O) m -CH2-CH2-heteroarylene-(CH2) p -,-(CH2CH 20) m -CH2-CH2-Cycloalkanediyl-(CH2) p -,-(CH2) n - Arirene-CH2-CH2-(OCH2CH2) m -,-(CH2) n -heterogeneous Ren-CH2-CH2-(OCH2CH2) m -, or -(CH2) n - Cyclo Alka Njiil-CH2-CH2-(OCH2CH2) m - represents. For more details, see Arreren phenylene is phenylene; cycloalkanediyl is cyclohexanediyl, for example, para -Cyclohexanediyl; heteroarylene is divalent 1,2,3-triazole, For example, divalent 1H-1,2,3-triazole.

[0067] According to a particular embodiment, L1' is -(CH2) n -,-(CH2CH2O) m -C H2-CH2-, Arirene, -Cycloalkanediyl-, -(CH2) n - Ariren -, -Ariren-(CH2) n -,-(CH2) n -Cycloalkanediyl-, -Sic Roalkandil-(CH2) n -, [ka] This represents... More specifically, arylene is phenylene; cycloalkanediyl is... Crohexanediyl, for example, p-cyclohexanediyl.

[0068] According to another specific embodiment, L1' is -(CH2) n -or-(CH2CH2O ) m -CH2-CH2-, especially -(CH2) n - represents, for example, -(CH2)5-.

[0069] The L1' stretcher unit section ends with the stretcher unit section CO-Z'. This can be done, and if z'=1, then Z' is -CO-NR4-(CH2) u -NR5 -or -CO-NR4-(CH2) u -NR5-CO-(CH2) v - is R4 R5 is independently H or (C1-C6) alkyl, for example, H or Me. and v are independently integers from 1 to 6, for example, from 1 to 4, in particular 1 or 2, for example 2. ru.

[0070] (W) w This represents the amino acid unit of the linker.

[0071] The amino acid units of the linker are not limited to releasing the drug, but It can be enzymatically cleaved by enzymes containing ulcer-associated proteases.

[0072] In terms of its selectivity for enzymatic cleavage by specific tumor-associated proteases, amino acid single The position can be designed and optimized. Suitable units are proteases, cathepsins, The cleavage is catalyzed by C and D, as well as plasmin.

[0073] (W) w It does not need to be present (w=0), or it can be a dipeptide, tripeptide, or tetrapeptide. It may be a peptide or pentapeptide unit (w=1, 2, 3, 4, or 5), Therefore, the amino acids that form a peptide can be different from each other.

[0074] Therefore, (W) w The formula is as follows: (W1) w1 (W2) w2 (W3) w3 (W4) w4 (W5 ) w5 It can be represented by, where each W1 to W5 independently represents an amino acid unit. Each of w1 to w5 is either 0 or 1.

[0075] In some embodiments, amino acid units (W) w These are naturally occurring substances, as well as Rare amino acids and unnatural amino acid analogs, such as citrulline, are used as amino acid residues. It can include.

[0076] amino acid unit (W) w The amino acid residues include, but are not limited to, alanine, b Phosphorus, leucine, isoleucine, methionine, phenylalanine, tryptophan, pro Phosphorus, acetyl or formyl-protected or unprotected lysine, arginine , arginine, histidine, protected or unprotected with tosyl or nitro groups, Ornithine, acetyl or formyl-protected or unprotected ornithine, It contains citrulline. A preferred example of an amino acid linker component is dipeptide. Contains cytoplasm or tripeptides.

[0077] Exemplary dipeptides include Val-Cit, Ala-Val, Ala-Ala, and Va l-Ala, Lys-Lys, Cit-Cit, Val-Lys, Ala-Phe, Ph e-Lys, Ala-Lys, Phe-Cit, Leu-Cit, Ile-Cit, Tr p-Cit, Phe-Ala, Phe-N 9 -Tosyl-Arg, and Phe-N 9 -Ni This includes Toro-Arg, preferably Val-Cit or Val-Ala.

[0078] Exemplary tripeptides include Val-Ala-Val, Ala-Asn-Val, and V al-Leu-Lys, Ala-Ala-Asn, Phe-Phe-Lys, Gly-G This includes ly-Gly, D-Phe-Phe-Lys, and Gly-Phe-Lys.

[0079] An example of a tetrapeptide is Gly-Phe-Leu-Gly (SEQ ID NO: 53). It contains Ala-Leu-Ala-Leu (sequence number 54).

[0080] An example of a pentapeptide is Pro-Val-Gly-Val-Val (SEQ ID NO:). 55) is included.

[0081] According to a particular embodiment, (W) w This is a dipeptide (i.e., w=2), for example, V al-Cit or Val-Ala, preferably Val-Cit, or ri The linker lacks an amino acid unit (w=0). In this case, preferably, it also lacks a spacer unit Y (y=0).

[0082] According to a preferred embodiment, w=0 (i.e., (W) w (is a single bond) or w =2 (that is, (W)) w (is a dipeptide), and therefore (W) w teeth, [ka] You can choose from, in particular, Val-Cit, and here, The asterisk represents the spacer unit (Y). y It shows the connection point with; and The dashed line represents -L1-(CO) c -(CO if c=1, or L1 if c=0) To indicate a point.

[0083] Y represents the spacer unit of the linker.

[0084] Spacer units are of two general types: self-destructive and non-self-destructive.

[0085] Non-self-destructing spacer units are formed after enzymatic cleavage of amino acid units from antibody-drug conjugates. Some or all of the spacer units remain bound to the drug. Non-self-destructing spacers Examples of spacer units include, but are not limited to, (glycine-glycine) spacers. —Includes units and glycine spacer units. To release the drug, glycine - Independent hydrolysis reactions must occur within the target cell to cleave the drug unit bond. No.

[0086] The self-destructing spacer unit can release drugs without the need for a separate hydrolysis process. In these embodiments, (Y) is a p-aminobenzyl alcohol unit (PAB) It is a residue, and via the nitrogen atom of the PAB group (W) w Linked to, ester group, carbonate group, It is directly attached to the drug via a rubamic acid group or ether group. It does not contain a PAB moiety. Such linkers can also be considered traceless linkers.

[0087] In this invention, the spacer unit (Y) is -PAB-CO-(Z) z -and here So, PAB is, [ka] (The oxygen in the PAB unit is linked to the carbonyl group), -para-aminobenzyl-O- Also referred to as CO-, and y=1, or this linker lacks spacer units. (y=0).

[0088] The spacer-para-aminobenzyl-O-CO- can be terminated with spacer Z. Spacer Z is -NR4-(CH2) when z=1. u -NR5- or -NR4- (CH2) u -NR5-CO- or even -NR4-(CH2) u -NR5-CO- (CH2) v -or -NR4-(CH2) u -NR5-CO-(CH2) v -CO- Yes. R4 and R5 are independently H or (C1-C6) alkyl, for example, H or M e. u and v are independent integers from 1 to 6, for example, 1 to 4, in particular 1 or 2, for example. For example, it's 2.

[0089] For the sake of advantage, if w is 0, then y is 0, and if w is an integer from 1 to 5, then y is 0 or The answer is 1, which means that the spacer unit Y can only exist if the amino acid unit W is present. It means that.

[0090] Preferably, when w is 0, y is 0, and when w is an integer from 1 to 5, y is 1. Therefore, the spacer unit Y is the amino acid unit (W). w If present, amino Unit (W) w If it does not exist, it means it does not exist.

[0091] According to a particular embodiment, the base-L1-(CO) c -(W) w -(Y) y - is, - (CH 2) n -,-(CH2) n -CO-, -(CH2CH2O) m -CH2-CH2-, -( CH2CH2O) m -CH2-CH2-CO-, -CH2-para-cyclohexyl-CO -, -aryl-(CH2) n -,-(CH2) n -Val-Cit-para-aminoben Jill-O-CO-,-(CH2) n -CO-Val-Cit-para-aminobenzyl-O -CO-, -(CH2CH2O) m -CH2-CH2-Val-Cit-Para-Aminobe Nzil-O-CO-,-(CH2CH2O) m -CH2-CH2-CO-Val-Cit -para-aminobenzyl-O-CO-,-CH2-para-cyclohexyl-CO-Val -Cit-para-aminobenzyl-O-CO-,-aryl-(CH2) n -Val-C it-para-aminobenzyl-O-CO-,-aryl-CO-Val-Cit-para- aminobenzyl-O-CO-,-(CH2) n -Val-Ala-para-aminobenzyl -O-CO-, -(CH2) n -CO-Val-Ala-para-aminobenzyl-OC O-, -(CH2CH2O) m -CH2-CH2-Val-Ala-para-aminobenzyl Lu-O-CO-,-(CH2CH2O) m -CH2-CH2-CO-Val-Ala-Pa La-aminobenzyl-O-CO-,-CH2-para-cyclohexyl-CO-Val-A la-para-aminobenzyl-O-CO-,-aryl-(CH2) n -Val-Ala -para-aminobenzyl-O-CO-,-aryl-CO-Val-Ala-para-ami Nobenzyl-O-CO-,-(CH2) n -Val-Cit-para-aminobenzyl-O -CO-NH-(CH2) u -NH-, -(CH2) n -CO-Val-Cit-Para- aminobenzyl-O-CO-NH-(CH2) u -NH-, -(CH2CH2O) m -C H2-CH2-Val-Cit-para-aminobenzyl-O-CO-NH-(CH2) u -NH-, -(CH2CH2O) m -CH2-CH2-CO-Val-Cit-para-a Minobenzyl-O-CO-NH-(CH2) u -NH-,-CH2-para-cyclohexyl Lu-CO-Val-Cit-para-aminobenzyl-O-CO-NH-(CH2) u -N H-, -aryl-(CH2) n -Val-Cit-para-aminobenzyl-O-CO- NH-(CH2) u -NH-,-aryl-CO-Val-Cit-para-aminobenzyl Ru-O-CO-NH-(CH2) u -NH-, -(CH2) n -Val-Ala-Para- aminobenzyl-O-CO-NH-(CH2) u -NH-, -(CH2) n -CO-Va l-Ala-para-aminobenzyl-O-CO-NH-(CH2) u -NH-, -(CH 2CH2O) m -CH2-CH2-Val-Ala-para-aminobenzyl-O-CO- NH-(CH2) u -NH-, -(CH2CH2O) m -CH2-CH2-CO-Val -Ala-para-aminobenzyl-O-CO-NH-(CH2) u -NH-, -CH2- para-cyclohexyl-CO-Val-Ala-para-aminobenzyl-O-CO-NH -(CH2) u -NH-, -aryl-(CH2) n -Val-Ala-Para-Aminobe Nzil-O-CO-NH-(CH2) u -NH-, -aryl-CO-Val-Ala- para-aminobenzyl-O-CO-NH-(CH2) u -NH-, -(CH2) n -Va l-Cit-para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-C O-, -(CH2) n -CO-Val-Cit-para-aminobenzyl-O-CO-NC H3-(CH2) u -NCH3-CO-, -(CH2CH2O) m -CH2-CH2-V al-Cit-para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3- CO-, -(CH2CH2O) m -CH2-CH2-CO-Val-Cit-Para-Ami Nobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-, -CH2-para- Cyclohexyl-CO-Val-Cit-para-aminobenzyl-O-CO-NCH3- (CH2) u -NCH3-CO-, -aryl-(CH2) n -Val-Cit-Para- aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-, -aryl- CO-Val-Cit-para-aminobenzyl-O-CO-NCH3-(CH2) u -N CH3-CO-, -(CH2) n -Val-Ala-para-aminobenzyl-O-CO- NCH3-(CH2) u -NCH3-CO-, -(CH2) n -CO-Val-Ala- para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-,-(C H2CH2O) m -CH2-CH2-Val-Ala-para-aminobenzyl-O-CO -NCH3-(CH2) u -NCH3-CO-, -(CH2CH2O) m -CH2-CH 2-CO-Val-Ala-para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-,-CH2-para-cyclohexyl-CO-Val-Ala-para- aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-, -aryl- (CH2) n -Val-Ala-para-aminobenzyl-O-CO-NCH3-(CH2 ) u -NCH3-CO-,-aryl-CO-Val-Ala-para-aminobenzyl- O-CO-NCH3-(CH2) u -NCH3-CO-, [ka] JPEG0007830555000033.jpg147144 JPEG0007830555000034.jpg154136 This represents JPEG0007830555000035.jpg118148.

[0092] Base-L1-(CO) c -(W) w -(Y) y -Also,-(CH2) n -CO-NCH 3-(CH2) u -NCH3-CO-(CH2) v -,-(CH2) n -CO-NCH3 -(CH2) u -NCH3-CO-(CH2) v -CO-, -(CH2CH2O) m -C H2-CH2-CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -,-( CH2CH2O) m -CH2-CH2-CO-NCH3-(CH2) u -NCH3-CO -(CH2) v -CO-,-CH2-para-cyclohexyl-CO-NCH3-(CH2 ) u -NCH3-CO-(CH2) v -,-CH2-para-cyclohexyl-CO-NC H3-(CH2) u -NCH3-CO-(CH2) v -CO-, -aryl-(CH2) n -CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -, -Aryl-( CH2) n -CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -CO-, -(CH2) n -CO-NCH3-(CH2) u -NCH3-, -(CH2) n -CO- NCH3-(CH2) u -NCH3-CO-, -(CH2CH2O) m -CH2-CH2 -CO-NCH3-(CH2) u -NCH3-, -(CH2CH2O) m -CH2-CH 2-CO-NCH3-(CH2) u -NCH3-, -(CH2CH2O) m -CH2-C H2-CO-NCH3-(CH2) u -NCH3-CO-,-CH2-para-cyclohex Sil-CO-NCH3-(CH2) u -NCH3-,-CH2-para-cyclohexyl- CO-NCH3-(CH2) u -NCH3-CO-, -aryl-(CH2) n -CO- NCH3-(CH2) u -NCH3-, -aryl-(CH2) n -CO-NCH3-( CH2) u -NCH3-, -aryl-(CH2) n -CO-NCH3-(CH2) u - NCH3-CO-,-(CH2) n -CO-NH-(CH2) u -NH-, -(CH2) n -CO-NH-(CH2) u -NH-CO-, -(CH2CH2O) m -CH2-CH 2-CO-NH-(CH2) u -NH-, -(CH2CH2O) m -CH2-CH2-C O-NH-(CH2) u -NH-CO-,-CH2-para-cyclohexyl-CO-NH -(CH2) u -NH-,-CH2-para-cyclohexyl-CO-NH-(CH2) u -NH-CO-, -aryl-(CH2) n -CO-NH-(CH2) u -NH-, -A Reel (CH2) n -CO-NH-(CH2) u -NH-CO-, -(CH2) n -V al-Cit-para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3- CO-(CH2) v -,-(CH2) n -Val-Cit-para-aminobenzyl-O- CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -CO-, -(CH2) n -CO-Val-Cit-para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -,-(CH2) n -CO-Val-Cit-para-a Minobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -C O-, -(CH2CH2O) m -CH2-CH2-Val-Cit-para-aminobenzyl Ru-O-CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -,-(CH2 CH2O) m -CH2-CH2-Val-Cit-para-aminobenzyl-O-CO-N CH3-(CH2) u -NCH3-CO-(CH2) v -CO-, -(CH2CH2O) m -CH2-CH2-CO-Val-Cit-para-aminobenzyl-O-CO-NCH 3-(CH2) u -NCH3-CO-(CH2)v -,-(CH2CH2O) m -CH2 -CH2-CO-Val-Cit-para-aminobenzyl-O-CO-NCH3-(CH 2) u -NCH3-CO-(CH2) v -CO-,-CH2-para-cyclohexyl-C O-Val-Cit-para-aminobenzyl-O-CO-NCH3-(CH2) u -NC H3-CO-(CH2) v -,-CH2-para-cyclohexyl-CO-Val-Cit -para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-(CH 2) v -CO-, -aryl-(CH2) n -Val-Cit-para-aminobenzyl- O-CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -, -Aryl-( CH2) n -Val-Cit-para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -CO-, -aryl-CO-Val-Cit-para -aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -,-aryl-CO-Val-Cit-para-aminobenzyl-O-CO-NCH3- (CH2) u -NCH3-CO-(CH2) v -CO-, -(CH2) n -Val-Al α-para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-(C H2) v -,-(CH2) n-Val-Ala-para-aminobenzyl-O-CO-NC H3-(CH2) u -NCH3-CO-(CH2) v -CO-, -(CH2) n -CO- Val-Ala-para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3 -CO-(CH2) v -,-(CH2) n -CO-Val-Ala-para-aminobenzyl Ru-O-CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -CO-, -( CH2CH2O) m -CH2-CH2-Val-Ala-para-aminobenzyl-OC O-NCH3-(CH2) u -NCH3-CO-(CH2) v -,-(CH2CH2O) m -CH2-CH2-Val-Ala-para-aminobenzyl-O-CO-NCH3-( CH2) u -NCH3-CO-(CH2) v -CO-, -(CH2CH2O) m -CH2 -CH2-CO-Val-Ala-para-aminobenzyl-O-CO-NCH3-(CH 2) u -NCH3-CO-(CH2) v -,-(CH2CH2O) m -CH2-CH2- CO-Val-Ala-para-aminobenzyl-O-CO-NCH3-(CH2) u -N CH3-CO-(CH2) v -CO-,-CH2-para-cyclohexyl-CO-Val -Ala-para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO -(CH2) v-, -CH2-para-cyclohexyl-CO-Val-Ala-para-a Minobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -C O-,-aryl-(CH2) n -Val-Ala-para-aminobenzyl-O-CO- NCH3-(CH2) u -NCH3-CO-(CH2) v -, -aryl-(CH2) n -Val-Ala-para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH 3-CO-(CH2) v -CO-,-aryl-CO-Val-Ala-para-aminobe Nzil-O-CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -or- aryl-CO-Val-A la-para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-( CH2) v -CO- can also be represented.

[0093] According to another specific embodiment, the group -L1-(CO) c -(W) w -(Y) y -teeth,-( CH2) n -,-(CH2) n -CO-, -(CH2) n -Val-Cit-Para-Ami Nobenzyl-O-CO-,-(CH2) n -CO-Val-Cit-para-aminobenz Ru-O-CO-, -(CH2) n -Val-Ala-para-aminobenzyl-O-CO- ,-(CH2) n -CO-Val-Ala-para-aminobenzyl-O-CO-,-(C H2) n -Val-Cit-para-aminobenzyl-O-CO-NH-(CH2) u -N H-, -(CH2) n -CO-Val-Cit-para-aminobenzyl-O-CO-NH -(CH2) u -NH-, -(CH2) n -Val-Ala-para-aminobenzyl-O -CO-NH-(CH2) u -NH-, -(CH2) n -CO-Val-Ala-Para- aminobenzyl-O-CO-NH-(CH2) u -NH-, -(CH2) n -Val-C it-para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-, -(CH2) n -CO-Val-Cit-para-aminobenzyl-O-CO-NCH3- (CH2) u -NCH3-CO-, -(CH2) n -Val-Ala-para-aminoben Zil-O-CO-NCH3-(CH2) u -NCH3-CO-, or -(CH2) n - CO-Val-Ala-para-aminobenzyl-O-CO-NCH3-(CH2) u -N This represents CH3-CO-, where n and u are as previously defined, and in particular, n=5 And u = 2.

[0094] Base-L1-(CO) c -(W) w -(Y) y -Also,-(CH2) n -CO-NH- (CH2) u -NH-, -(CH2) n -CO-NH-(CH2) u-NH-CO-,- (CH2) n -CO-NCH3-(CH2) u -NCH3-, -(CH2) n -CO-N CH3-(CH2) u -NCH3-CO-, -(CH2) n -CO-NCH3-(CH2 ) u -NCH3-CO-(CH2) v -,-(CH2) n -CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -CO-, -(CH2) n -Val-Cit-Para- aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-(CH2) v - ,-(CH2) n -Val-Cit-para-aminobenzyl-O-CO-NCH3-(C H2) u -NCH3-CO-(CH2) v -CO-, -(CH2) n -CO-Val-C it-para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-( CH2) v -,-(CH2) n -CO-Val-Cit-para-aminobenzyl-OC O-NCH3-(CH2) u -NCH3-CO-(CH2) v -CO-, -(CH2) n -Val-Ala-para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH 3-CO-(CH2) v -,-(CH2) n -Val-Ala-para-aminobenzyl- O-CO-NCH3-(CH2) u-NCH3-CO-(CH2) v -CO-, -(CH 2) n -CO-Val-Ala-para-aminobenzyl-O-CO-NCH3-(CH2 ) u -NCH3-CO-(CH2) v -, or -(CH2) n -CO-Val-Ala -para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-(CH 2) v -CO- can also be represented, where n, u, and v are as previously defined, in particular, n=5 and u=v=2.

[0095] According to a preferred embodiment, the group -L1-(CO) c -(W) w -(Y) y - is, -(C H2) n -CO-, -(CH2) n -CO-Val-Cit-para-aminobenzyl-O -CO-, -(CH2) n -CO-Val-Ala-para-aminobenzyl-O-CO- ,-(CH2) n -CO-Val-Cit-para-aminobenzyl-O-CO-NH-( CH2) u -NH-, -(CH2) n -CO-Val-Ala-para-aminobenzyl- O-CO-NH-(CH2) u -NH-, -(CH2) n -CO-Val-Cit-para -aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-, or -( CH2) n -CO-Val-Ala-para-aminobenzyl-O-CO-NCH3-(C H2) uThis represents -NCH3-CO-, where n and u are as previously defined. In particular, n=5 and u=2.

[0096] Base-L1-(CO) c -(W) w -(Y) y -Also,-(CH2) n -CO-NH- (CH2) u -NH-, -(CH2) n -CO-NH-(CH2) u -NH-CO-,- (CH2) n -CO-NCH3-(CH2) u -NCH3-, -(CH2) n -CO-N CH3-(CH2) u -NCH3-CO-, -(CH2) n -CO-NCH3-(CH2 ) u -NCH3-CO-(CH2) v -,-(CH2) n -CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -CO-, -(CH2) n -CO-Val-Cit- para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-(CH2 ) v -,-(CH2) n -CO-Val-Cit-para-aminobenzyl-O-CO-N CH3-(CH2) u -NCH3-CO-(CH2) v -CO-, -(CH2) n -CO -Val-Ala-para-aminobenzyl-O-CO-NCH3-(CH2) u -NCH 3-CO-(CH2) v -, or -(CH2) n -CO-Val-Ala-Para-Ami Nobenzyl-O-CO-NCH3-(CH2) u -NCH3-CO-(CH2) v -CO - can represent, where n, u and v are as previously defined, in particular n=5 and u=v=2.

[0097] base [ka] Preferably, [ka] It reacts with a binding unit such as an antibody, and the sulfhydryl group present on the binding unit And it is a functional group that attaches the drug portion to it. The sulfhydryl group, when present, is particularly It can be produced by reducing the intramolecular disulfide bond of the antibody binding unit. Alternatively, the sulfhydryl group converts the amino group of the lysine portion of the bonding unit to 2-iminothiolane. Alternatively, it can be produced by reacting it with other sulfhydryl-producing reagents. In certain embodiments, the binding unit, such as an antibody, is manipulated to have one or more lysines. More preferably, the binding unit, such as an antibody, is manipulated to have the above-mentioned cysteine ​​groups. It is possible (compared to ThioMabs).

[0098] X1 and X2 are H, a halogen atom (e.g., Cl or Br), ( C1-C6) alkoxy, optionally substituted aryloxy, or -O -(CH2CH2O) r It represents H, however X1 and X2 cannot represent H at the same time. . Aryloxy is more specifically a halogen, CN, NO2 and aryloxy (for example) (If phenyloxy) may be substituted with one or more halogen atoms such as fluorine atoms. (May be) optionally replaced by one or more (e.g., one) bases selected from It may be done. In particular, aryloxy is CN, NO2 and pentafluorophenyl One or more selected from luoxy (in particular, which may be optionally substituted with CN) It may be optionally substituted with (for example, one) group. Aryloxy is particularly, It can be an phenyloxy.

[0099] According to a particular embodiment, X1 and X2 are independent of each other, H, halogen atoms (for example) If it is substituted with Cl or Br, (C1-C6) alkoxy or optionally It represents an aryloxy, however X1 and X2 cannot simultaneously represent H. More specifically, aryloxys are halogens, CN, NO2, and aryloxys (e.g., fe (Nyloxy) (occasionally substituted with one or more halogen atoms such as a fluorine atom) (May be) Optionally replaced by one or more (e.g., one) bases selected from It may also be the case that aryloxys are CN, NO2 and pentafluorophenyloxy. One or more selected from (in particular, which may be replaced in some cases with CN) (e.g.) It may be optionally substituted with one group. Aryloxy is particularly phenyloxy It could be Kishi.

[0100] According to another specific embodiment, X1 and X2 are independent of each other, H, Cl, Br, M Toxy, or phenyloxy substituted with CN, in particular representing H, Cl or Br Furthermore, X1 and X2 cannot simultaneously represent H.

[0101] Advantageously, X1 and X2 are identical and not H, or one of X1 and X2 One is H, and the other is not H. If X1 and / or X2 are not H, then it is Cl. Alternatively, halogen atoms such as Br, (C1-C6) alkoxys, and in some cases, even if substituted. Good aryloxy, or -O-(CH2CH2O) r H; especially Cl or Br, etc. The halogen atom, (C1-C6) alkoxy or optionally substituted Phenyloxy; preferably phenyloxy substituted with Cl, Br, methoxy or CN In particular, it is Cl or Br.

[0102] q represents 0, 1, or 2. Preferably, q represents 2.

[0103] X3 is used to covalently link the drug to a binding unit such as an antibody, and the drug (Q) A functional group intended to react with H or Q-OH (in the case of y=z=1, depending on the case) The terminal nitrogen of Z is -NR4-(CH2) u - If it is NR5-, then Z is -NR4-(CH2) u -NR5- or the end of Z' It represents (containing nitrogen).

[0104] Furthermore, before linking the stretcher units having a sulfomaleimide functional group, In this case, the spacer unit Y and the amino acid unit (W) w It is also conceivable to introduce it into the drug portion. It can be determined. In this case, w=y=0 (i.e., stretcher unit and sulfomalymi A compound of formula (I) (containing only functional groups) is used, where X3 is the drug unit in this case. Amino acid units (W) already bound to it wAlternatively, a sensory component that reacts with spacer unit Y. It represents the basis.

[0105] y=z=1 and Z is -NR4-(CH2) u -NR5- or c=w=y= 0, z'=1 and Z' is -NR4-(CH2) u -NR5- (and Z or Z') X3 represents H when it forms an NH functional group with the terminal nitrogen of the group, and in other cases X3 represents OH, NH2, or a leaving group, for example, OH or a leaving group. The leaving group is NH2. Logen atoms (e.g., Cl, Br, I), sulfonates (e.g., OTf, OMs, OT) s), N-succinimidyloxy, 4-nitrophenyloxy, pentafluorophenyl It may be yloxy or N-benzotriazoloxy. In particular, y=z=1 and Z is -N R4-(CH2) u -NR5- or if c=w=y=0, z'=1 and Z' is - NR4-(CH2) u If it is -NR5-, then X3 represents H, otherwise X3 is More specifically, it may be OH, Cl, or N-succinimidyloxy.

[0106] Drug portion The drug portion (Q) is a residue of either drug QH or drug Q-OH.

[0107] The drug according to the present invention is useful for human therapy or veterinary therapy, particularly for the treatment of cancer. It could be any drug. The drug could be a cytotoxic agent in particular. Advantageously, such The drug comprises a functional group that allows the drug to be linked to the linker portion. Furthermore, it is conceivable that such functional groups may be added to the drug in order to achieve crosslinking. The functional group can be, for example, OH, SH, NH or COOH, and the drug can be linked to the linker portion. The X3 end of the linker is reacted to form a coupling reaction. Conversion (for example, reaction between OH, SH, NH or COOH and X3 = leaving group), peptide cuts Pulling (for example, COOH and ZX3 or Z with X3=NH2 or NH at the end) Possible reactions include (reaction with X3), esterification (reaction between COOH and OH), and the Mitsunobu reaction. .

[0108] The drug portion Q could be, for example, the following: Residues of auristatin derivatives, for example, residues of monomethyl auristatin (MMAF) Monomethyl auristatin E (linked by its terminal NH or COOH group) (MMAE) residues (linked by terminal NH or OH groups), monomethyl Residues of rastatin-10 (linked by their terminal NH groups) or derivatives thereof, e.g. For example, the residues of the drug portion of formula (C) as defined below;

[0109] [Table 1]

[0110] Anthracycline residues, such as daunorubicin, doxorubicin, and epirubicin. or idarubicin residues (-COCH2OH linked by the NH2 or OH group) (or its derivatives, e.g., 2-pyrrolidoxorubicin or pro-2-pyrro A residue of linodoxorubicin (linked by the OH group of -COCH2OH), or A residue of PNU-159682 (linked by the OH group of -COCH2OH) or The residues of its derivatives; in particular, the residues of doxorubicin (the NH2 group of -COCH2OH or (Linked by an OH group), 2-pyrrolidodoxorubicin, pro-2-pyrrolidodox Sorbicin residues (linked by the OH group of -COCH2OH) or PNU-1 59682 residues (linked by the OH group of -COCH2OH) or especially the following Residues of derivatives of PNU-159682 as shown; preferably PNU-1596 82 residues (linked by the OH group of -COCH2OH) or shown below Residues of PNU-159682 derivatives (linked by COOH);

[0111] [Table 2] JPEG0007830555000040.jpg175132

[0112] Camptothecin or its derivatives, for example, SN-38 residues (linked by their OH groups) Like this;

[0113] [Table 3]

[0114] Tubricin, for example, tubulicin A, tubulicin B, tubulicin C or tubulosin D residues (if present, linked by COOH or OH groups) They are there;

[0115] [Table 4]

[0116] Kalicheamicin, for example, esperamicin or kalicheamicin γ1, or so Derivatives of, for example, the residue of N-acetyldimethylhydrazidekalychaemycin (the hydr Connected by the radiated section);

[0117] [Table 5]

[0118] Maytansinoids, such as maytansine (also called maytansine) or its derivatives. For example, residues of DM1 or DM4 (linked by SH groups); in particular, DM1 or residues of DM4 (linked by SH groups);

[0119] [Table 6]

[0120] Duocalmycin, for example, duocalmycin A, duocalmycin B1, du Ocalmycin B2, Duocalmycin C1, Duocalmycin C2, Duocalmycin Isine D, duocalmycin SA, or residues of CC-1065 (by CONH2 group) Linked; in particular, the residue of CC-1065 (linked by two CONH groups) ;

[0121] [Table 7]

[0122] Amanitin residues (OH, NH, COOH or CONH2 groups, especially those linked by an OH group) (They are linked), for example, α-amanitin, β-amanitin, γ-amanitin or ε- Amanitin residues; in particular, α-amanitin residues (especially those linked by a CH2OH group) They are there;

[0123] [Table 8] JPEG0007830555000047.jpg195136

[0124] Pyrrolobenzodiazepines (PBDs), for example, the residues of anthramycin (and their OH groups) (is linked by an NH2 group) or residues of SGD-1882 (by its NH2 group) Linked; in particular, residues of SGD-1882 (linked by their NH2 groups) );

[0125] [Table 9]

[0126] The residues of the immune checkpoint activator, for example, are defined in the following formula: D) STING (interferon gene stimulant) agonist residues (OH, SH or (is linked by NH) or IDO (indoleamine 2,3-dioxygenate) ) Inhibitors, for example, epacadostat (INCB024360) or BMS-986 205 residues.

[0127] For the sake of advantage, drug portion Q is as follows: Residues of auristatin derivatives, for example, residues of MMAF (their terminal NH or COOH). (linked by a group), MMAE residues (linked by their terminal NH or OH groups) (is), or a residue of monomethyldrastatin-10 (linked by its terminal NH group) (The residues of the drug moiety of formula (C) as defined below) STING agonists, in particular residues of formula (D) as defined below; or Anthracycline residues such as those defined above, and preferably those shown below. Residues of PNU-159682 or its derivatives that are such as:

[0128] [Table 10]

[0129] Drug portion Q is, in particular, an anthracycline residue as defined above, and preferred The residues of PNU-159682 or its derivatives are as follows:

[0130] [Table 11]

[0131] According to the first embodiment, the residue of the auristatin derivative is given by the following formula (C): [ka] It has, in the formula, R1 is either H or OH. R2 is (C1-C6)alkyl (e.g., methyl), COOH, COO-((C1- C6) alkyl) (e.g., COOMe) or thiazolyl (e.g., thiazole-2- (Ill) R3 is H or (C1-C6) alkyl (e.g., methyl), in particular (C1-C6) It is an alkyl group, X4 is O or NR9, R9 is H or (C1-C6) alkyl (e.g., methyl), and t is an integer between 1 and 8, particularly between 1 and 6, preferably between 1 and 4, and is more preferably 1 or The answer is 2.

[0132] According to a particular embodiment, R1 is OH and R2 is (C1-C6) alkyl, for example, methyl. ;or R1 is H, and R2 is thiazolyl, e.g., thiazole-2-yl, COO-( C1-C6 alkyl groups, for example, COOMe or COOH.

[0133] Advantageously, R1 is H, and R2 is thiazolyl, for example, thiazole-2- yl, COO-(C1-C6) alkyl, for example, COOMe, or COOH. Preferably, R1 is H, and R2 is COOH or COOMe, in particular CO It is OH.

[0134] t is an integer between 1 and 8, particularly between 1 and 6, preferably between 1 and 4, and is more preferably 1 or The answer is 2.

[0135] Advantageously, R3 is a (C1-C6) alkyl group, preferably a methyl group.

[0136] According to a particular embodiment, R1 is H, and R2 is COOH or COOMe (preferably) The compound is COOH, R3 is methyl, and t is 1 or 2.

[0137] Advantageously, X4 is NR9, where R9 is H or (C1-C6) alkyl. Preferably, it is H or methyl.

[0138] In a preferred embodiment, R1 is H, R2 is COOH, R3 is methyl, and X4 is NR9. R9 is methyl, and t is 1 or 2, or R1 is H, R2 is COOH, R3 is methyl, and X4 is NR9. R9 is H, and t is either 1 or 2.

[0139] According to a preferred embodiment, the X4 group is (CH2) on the phenyl ring. t Para with respect to the base It is in a certain position.

[0140] Advantageously, the auristatin residues in formula (C) are selected from the following parts. [ka] JPEG0007830555000053.jpg221140 JPEG0007830555000054.jpg43136

[0141] The preparation of such auristatin derivatives can be seen, for example, in WO2014 / 174064. This is disclosed in WO2015 / 162293.

[0142] According to the second embodiment, the STING agonist is given by the following equation (X): [ka] It has, in the formula, X 11 and X 21 Independently, O or S, preferably O, X 12 and X 22 These are independently OH or SH, preferably SH. A 11 and A 21 These are independently, formula: [ka] Preferably, [ka] It is the basis of, and here, • Z1 is OR 11 or NR 11 R 12 And here, R 11 and R 12 Independently H, R 13 or COR 13 And here, R 13 (C1-C6) alkyl, A It is a reel or aryl(C1-C6)alkyl, • Z2 is H or NR 21 R 22 And here, R 21 and R 22 H, independently R 23 or COR 23 And here, R 23 These are (C1-C6) alkyl and aryl compounds. or aryl(C1-C6)alkyl, • Z3 is N or CR 33 Preferably, N, where R 33 is H or halo A gen atom, for example, F or Cl, and • Z4 is H or (C1-C6) alkyl, A 12 and A 22 These are independently H, OH, or F, and A2 is H, or A2 and A 22 They are interconnected, where A2 is connected to CH2. Yes, A 22 It is O.

[0143] If Z1 is OH or Z4 is H, the following tautomers are obtained. [ka]

[0144] According to a particular embodiment, the STING agonist has one of the following formulas: [ka] (In the formula, X 11 , X 21 , X 12 , X 22 , A 11 , A 21 , A 12 , A 22 and A2 (As defined above or below).

[0145] According to another specific embodiment, the STING agonist has one of the following formulas: [ka] (In the formula, X 11 , X 21 , X 12 , X 22 , A 11 , A 21 , A 12 , A 22 and A2 (As defined above or below).

[0146] To have an advantage, X 11 and X 21 Both are O. To be advantageous, X 12 and X 22 of At least one is SH, preferably X 12 and X 22 Both are SH. Preferably, X 11 and X 21 Both are O, and X 12 and X 22 Both Both are SH.

[0147] In particular, R 11 and R 12 Both are H, and to the advantage, R 11 , R 12 , R 21 oh Call R 22These are H.

[0148] Z3 is specifically N. Advantageously, Z1 is OH or NH2; Z2 is or N H2 is N; Z3 is N; and Z4 is H.

[0149] Preferably, A 11 and A 21 Independently, [ka] Selected from, more preferably cytosine, adenine, adenine-6-benzamide, 2 ,6-diaminopurine, hypoxanthine, guanine and guanine-2-isobutyramide Selected from; most preferably selected from adenine, hypoxanthine and guanine. ru.

[0150] This could be the ADU-S100, shown below. [ka]

[0151] Alternatively, see Lioux et al. J. Med. Chem., 2016, 59 (22), pp 10253-102. It may be one of the compounds specifically disclosed in 67.

[0152] The fabrication of such STING agonists is, for example, WO2014 / 179335, W O2016 / 096174, WO2016 / 145102, WO2017 / 106740 Alternatively, it is disclosed in WO2018 / 100558.

[0153] The aforementioned STING agonist, due to the SH, OH, or NH groups present in the molecule, Chi, base X 12 (OH or SH), X22 (OH or SH), R 11 and R 12 few Even if one of them is H, then Z1(OH, NHR) 11 or NHR 12 ), or R 21 and R 22 If at least one of them is H, then Z2(NHR 21 or NHR2 2) It is connected to the linker portion. Preferably, X 12 and X 22 at least The other side is an SH group, and the STING agonist is linked by this SH group.

[0154] In conclusion, the residues of the STING agonist are favorably expressed in the following equations (D), (D-1), ( D-2), (D-3), (D-1a), (D-2a), or (D-3a): [ka] It has, in the formula, X 11 and X 21 As defined above, X 12 and X 22 As defined above, or O or S, A 11 and A 21 As defined above, that is, independently of the formula: [ka] Based on, preferably, [ka] And in the formula, Z1 is as defined above, or O or NR 11 And, • Z2 is as defined above, or NR 21 And, • Z3 is as defined above, and The Z4 is as defined above, A 12 and A 22 It is as defined above, and A2 is as defined above, Here, ·X 12 If O or S, then X 22 is not O, and is not S, and Z1 is not O , and NR 11 Not O, and not S, Z1 is not O, and NR 11 Instead, the residue of the STING agonist is X 12 The rest of the molecule is linked by Ori; ·X 22 If O or S, then X 12 is not O, and is not S, and Z1 is not O , and NR 11 Not O, and not S, Z1 is not O, and NR 11 Instead, the residue of the STING agonist is X 22 The rest of the molecule is linked by Ori; • Z1 is O or NR 11 If X 12 It is not O, and not S, but X 22 is O Not and not S, not O, and not S, Z1 is not O, and NR 11 Instead, the STING agonist residue is linked to the rest of the molecule by Z1. the law of nature; • Z2 is NR 21 If X 12 It is not O, and not S, but X 22 It is not O, Furthermore, S is not, Z1 is not O, and NR 11 Instead, the residues of the STING agonist It is linked to the rest of the molecule by Z2.

[0155] Joining unit The binding units are peptides, proteins (e.g., manipulated proteins), antibodies (e.g., mono- It is a clonal antibody or its antigen-binding fragment.

[0156] Preferably, the binding unit according to the present invention is an antibody or its antigen-binding fragment. Therefore, the binding unit drug conjugate according to the present invention is an antibody-drug conjugate (ADC). In the application form, the antibody of the present invention consists of a recombinant antibody. In another embodiment, the present invention The antibodies in ADCs consist of chemically synthesized antibodies.

[0157] More specifically, such molecules are interconnected by disulfide bonds, at least It consists of a glycoprotein comprising two heavy (H) chains and two light (L) chains. Each heavy chain is heavy Chain variable region (or domain) (abbreviated herein as HCVR or VH) and weight It comprises a chain-steady region. The heavy chain-steady region consists of three domains CH1, CH2, and CH3 It comprises the following: Each light chain has a light chain variable region (hereinafter abbreviated as LCVR or VL) and comprises a light chain constant region. The light chain constant region comprises one domain CL. The VH and VL regions are further subdivided into hypervariable regions called complementarity-determining regions (CDRs). This allows for the insertion of a more conserved area called the Framework Region (FR). Each VH and VL consists of three CDRs and four FRs, starting from the amino terminus. The following order is arranged towards the carboxyl terminus: FR1, CDR1, FR2, CDR2 FR3, CDR3, FR4. Variable regions of the heavy and light chains are binding domains that interact with the antigen. Includes the main cell. The constant region of the antibody is found in various cells of the immune system (e.g., effector cells). Immunoglobulins to host tissues or factors containing the first component of the classical complement system (Clq) It can mediate the binding of.

[0158] In one embodiment, the "antigen-binding fragment" is Fv, scFv (sc is single-stranded). (represents), Fab, F(ab')2, Fab', scFv-Fc fragment or die Abody, or poly(alkylene) glycol, for example, poly(ethylene) glycol Addition of 'PEGylation' (Fv-PEG, scFv-PEG, Fab-PEG, F(a b') PEGylated fragments called 2-PEG or Fab'-PEG) ("PE Through chemical modifications such as "G" (which stands for poly(ethylene) glycol), or into liposomes Selected from a group consisting of any fragments whose half-life is extended by the incorporation of, The aforementioned fragment has at least one of the characteristic CDRs of the antibody according to the present invention. Preferably, the "antigen-binding fragments" are the variable heavy chains or of the antibodies from which they are derived. It is composed of a partial arrangement of variable light chains, or comprises the partial arrangement said to be the partial arrangement It has the same binding specificity to the target as the antibody from which it was derived, and preferably it is derived At least 1 / 100 of the affinity of the generated antibody, more preferably at least 1 / 1 It is sufficient to maintain sufficient affinity equivalent to 0. More preferably, the "antigen binding" A "fragment" is at least three CDRs of the variable heavy chain of the antibody from which they originate, in other words, The three CDRs are CDR-H1, CDR-H2, and CDR-H3, and the variable light chain, in other words It consists of CDR-L1, CDR-L2 and CDR-L3, or they It includes.

[0159] According to a preferred embodiment, the binding unit is an IGF-1R antibody, a HER2 antibody or the It is an antigen-binding fragment.

[0160] The HER2 antibody, more specifically, is trastuzumab.

[0161] In one embodiment of the present invention, the antibody is an IGF-1R antibody, and the epitope of the antibody is Prioritizing the extracellular domain of human IGF-1R (also known as IGF-1R ECD) It is located in ).

[0162] In a particular embodiment, the antibody, or any antigen-binding fragment thereof, is 10 × 1 0 -10 ~1 × 10 -10 , more importantly, 8x10 -10 ~2×10 -10 Includes between Made EC 50 And it can bind to IGF-1R.

[0163] Competition for binding with IGF-1R is not limited, but radioactivity, Biac Any method or technique known to those skilled in the art, such as ORE, ELISA, or flow cytometry. This can be determined by the following: If there is "competition for binding with IGF-1R", At least 20%, with priority at least 50%, and with even higher priority at least 70% This means a percentage of competition.

[0164] The determination of binding to the same epitope is not limited, but radioactivity, Biacor e, ELISA, flow cytometry, and any other method or technique known to those skilled in the art This can be done. If it "binds to the same epitope as IGF-1R", it is less At least 20%, preferentially at least 50%, and more preferentially at least 70% of the competition It means "to combine".

[0165] As described above, in contrast to common knowledge, the present invention provides internal transfer after IGF-1R binding. We focus on specific IGF-1R antibodies that exhibit high efficacy. In this case, it is "to be internally migrated" or "has been internally migrated" (these two expressions are equivalent). Antibodies are taken up by mammalian cells when they bind to IGF-1R on the cell surface. This means that it enters the body. Such antibodies are of interest as part of ADCs. Therefore, it addresses linked cytotoxicity to target cancer cells, and It is directed towards the cancer cells. Once it has been internalized, its cytotoxic properties induce the death of cancer cells.

[0166] Advantageously, the IGF-1R antibody according to the present invention is effective against CDR-H2, CDR-H3, and CD All R-L2 sequences are identical, while the other three CDRs are different. Antibody binding specificity. Regarding this, CDR-H3 is described as being the most important and most relevant to epitope recognition. Since this is considered common knowledge, this observation appears to be consistent.

[0167] A key factor in the success of ADC therapy is the target antigen specificity and the internalization of the antigen-antibody complex into cancer cells. It appears to be a partial migration. Antigens that clearly do not migrate internally are more likely to migrate internally than antigens that do migrate internally. It has a low effectiveness in delivering damaging drugs. The internal distribution process differs depending on the antigen, and by antibody It depends on multiple parameters that can be affected.

[0168] In ADCs, the drug portion confers cytotoxic activity, and the antibody used is specifically targeted at cancer cells. In addition to fulfilling the role of the opposite sex, it also enters the cell in order to correctly address cytotoxicity. It is a vector. Therefore, in order to improve ADC, the antibody is used to increase the intracellular activity of target cancer cells. It may exhibit translocation ability. The efficiency of antibody-mediated intracellular translocation depends on the targeted epitope. They are significantly different. The selection of a potent internally permeable IGF-1R antibody is important for downregulating IGF-1R. We are conducting various experiments to study not only the nodes but also the subsequent internal transfer of IGF-1R antibodies to cells. It requires a Ta.

[0169] In one embodiment, the internal transfer of antibodies in the ADC according to the present invention is performed by immunofluorescence or F ACS (flow cytometry) (as illustrated below in this application) or internal migration mechanism It can be evaluated by any method or process known to those skilled in the art. In a preferred embodiment, the antibody of the ADC according to the present invention is at least 30%, preferentially It induces internal translocation after binding to 50% and more preferably 80% of IGF-1R. obtain.

[0170] The IGF-1R / antibody complex undergoes internal migration after the antibody binds to the ECD of IGF-1R. This process induces a decrease in the amount of IGF-1R on the cell surface. This decrease is not limited to specific examples. Any of the following known to those skilled in the art, such as Western blotting, FACS, and immunofluorescence It can be quantified by the following method.

[0171] In one embodiment, this reduction, which in turn reflects internal migration, is preferably FA Measured by CS, average at 4°C after 4 hours of incubation with antibody. It can be expressed as the difference or Δ between the fluorescence intensity (MFI) and the MFI measured at 37°C. Cut.

[0172] As an example without limitation, this Δ is i) a 4-hour incubation with the antibody described herein. ii) Secondary antimicrobial cells labeled with Alexa488 after the IV period The MFI is determined using whole-body samples, based on the MFI obtained from untreated cells and antibody-treated cells. This parameter is given by the following formula: Δ(MFI 4℃ -MFI 37℃ It is defined as being calculated using ). .

[0173] The difference between these MFIs is proportional to the amount of IGF-1R expressed on the cell surface. This reflects the downward adjustment of IGF-1R.

[0174] In one advantageous aspect, the antibody has a Δ(MFI) effect against MCF-7. 4℃ -MFI 37℃ The antibody comprises at least 280, preferably at least 400, antibodies that induce )

[0175] More specifically, the above-mentioned Δ can be measured according to the following steps, but these are examples. This must be considered an illustrative and unrestricted example: a) In complete culture medium at either cold (4°C) or warm (37°C) the target tumor cells. , treat and incubate with the antibody of the present invention; b) Treat the treated cells from step a) and, in parallel, treat the untreated cells with a secondary antibody; c) MFI (typical amount of IGF-1R present on the surface) of treated and untreated cells , measured using a secondary labeled antibody capable of binding to the antibody of the present invention; and d) The difference between the MFI obtained from untreated cells and the MFI obtained from treated cells. Then calculate Δ.

[0176] From this ΔMFI, the internal transition percentage is calculated. 100×(MFI 4℃ -MFI 37℃ ) / MFI 4℃ It can be calculated as follows.

[0177] The antibody for ADC according to the present invention is preferably 50% to 99% of MCF7, 70% This shows the internal migration percentage that falls between % and 90%, with a preference for the 75% to 87% range.

[0178] The specific advantages of the antibodies described herein are due to their internal transfer rates.

[0179] In general, with regard to ADCs, the antibody used should preferably be administered within 24 hours of the antibody administration. A rapid internal transition is preferably achieved within 12 hours, and even more preferably within 6 hours. It is known that demonstrating this is desirable.

[0180] In the present invention, the internal migration rate is also referred to as the decrease in cell surface-bound antibodies or the attenuation of cell surface antibodies. This is expressed as t1 / 2 (half-life), which is used to obtain a 50% reduction in ΔMFI. This corresponds to the required time (this aspect will be clearly understood in the following example).

[0181] A particular advantage is that the antibody of the ADC of the present invention is contained between 5 and 25 minutes, preferably between 10 and 20 minutes. This means that it has t1 / 2.

[0182] According to a particular embodiment of the present invention, the antibody is a three-fold hemisphere of the sequences of SEQ ID NOs: 1, 2, and 3 It comprises a chain CDR and three light chain CDRs of the sequences of sequence numbers 4, 5, and 6.

[0183] According to a particular embodiment of the present invention, the antibody is a sequence of SEQ ID NOs: 1, 2, and 3, or a sequence of SEQ ID NOs: 1, 2, and 3. Column numbers 1, 2, or 3 and at least 80%, preferably 85%, 90%, 95%, and Three heavy-stranded CDRs comprising or consisting of any sequence exhibiting 98% identity; The sequence of sequence numbers 4, 5, and 6, or sequence numbers 4, 5, or 6 and at least 80% preferably does not include any sequence exhibiting 85%, 90%, 95%, and 98% identity. It comprises three light chain CDRs consisting of or .

[0184] According to a particular embodiment of the present invention, the bonding unit is i) Having CDR-H2 of sequence number 2 and CDR-H3 of sequence number 3 Three heavy chain CDRs, and three light chain CDRs having CDR-L2 in the sequence of SEQ ID NO: 5 An antibody comprising the components of; ii) Antibodies that compete with the antibody in i) for binding to IGF-1R; and iii) An antibody that binds to the same epitope as the antibody in i) for IGF-1R. An antibody capable of binding to human IGF-1R, or its antigen-binding fragment, selected from the above. That is the case.

[0185] According to a particular embodiment of the present invention, the bonding unit is i) Three heavy chain CDRs of sequences 1, 2 and 3, as well as sequences 4 and 5 An antibody comprising three light chain CDRs of sequence 6; ii) Antibodies that compete with the antibody in i) for binding to IGF-1R; and iii) An antibody that binds to the same epitope as the antibody in i) for IGF-1R. An antibody capable of binding to human IGF-1R, or its antigen-binding fragment, selected from the above. That is the case.

[0186] In another embodiment, the antibody, or its antigen-binding fragment, is SEQ ID NOs. 1, 2 Three heavy chain CDRs comprising sequences 4, 5 and 6; and sequences 4, 5 and 6 It contains three light chain CDRs.

[0187] IMGT's unique numbering system is variable regardless of the antigen receptor, chain type, or species. This was defined for comparison with the main [Lefranc M.-P., Immunology Today 18, 5 09 (1997) / Lefranc M.-P., The Immunologist, 7, 132-136 (1999) / Lefranc, M.-P., Pommie, C., Ruiz, M., Giudicelli, V., Foulquier, E., Truong, L., Thouvenin-Cont. et, V. and Lefranc, Dev. Comp. Immunol., 27, 55-77 (2003)]. IMGT's unique numbering. For example, in the case of cysteine ​​23 (1st-CYS) and tryptophan 41 (CONS) ERVED-TRP), hydrophobic amino acid 89, cysteine ​​104 (2nd-CYS), f Preservatives such as phenylalanine or tryptophan 118 (J-PHE or J-TRP) The amino acids always occupy the same position. IMGT's unique numbering is based on the framework. - Area (FR1-IMGT: 1~26, FR2-IMGT: 39~55, FR3- IMGT: 66-104 and FR4-IMGT: 118-128) and complementary resolution Fixed area: CDR1-IMGT: 27-38, CDR2-IMGT: 56-65 and Provides standard demarcation for CDR3-IMGT:105~117. The gaps are occupied. This indicates the position that is not present, so the CDR-IMGT length is shown separated by dots in parentheses. For example, [8.8.13]) is important information. IMGT's unique numbering is I MGT Colliers de Perles [Ruiz, M. and Lefranc, M.-P., Immuno genetics, 53, 857-883 (2002) / Kaas, Q. and Lefranc, M.-P., Current Bioinformati 2D graphs and IMGT / 3D structures represented as cs, 2, 21-30 (2007) e-DB[Kaas, Q., Ruiz, M. and Lefranc, M.-P., T cell receptor and MHC structur It is used in the 3D structure in al data. Nucl. Acids. Res., 32, D208-D210 (2004).

[0188] Reference amino acid sequence and at least 80%, preferably 85%, 90%, 95%, and 98% For amino acid sequences showing % identity, preferred examples include reference sequences, specific modifications, and especially , including deletion, addition or substitution, terminal cleavage or extension of at least one amino acid This includes. In the case of substitution of one or more conserved or non-conserved amino acids, the substituted amino acids are "equivalent to". Substitutions that replace amino acids with "equivalent" amino acids are preferred. Here, the expression "equivalent amino acid" means Without altering the structure of the corresponding antibody and the specific example defined below, the structure This means that it indicates any amino acid that can be substituted for one of the target amino acids.

[0189] Equivalent amino acids have structural homology with the substituted amino acid or can produce various antibodies. This can be determined based on the results of comparative tests of biological activity between organisms.

[0190] As an example that is not limited to this, the modified antibodies corresponding to Table 1 below have a significant change in their biological activity. List the possible substitutions that can be performed without [the same conditions]; the reverse substitutions are, of course, also possible under the same conditions. .

[0191] [Table 12]

[0192] A particular aspect of the present invention is that the antibody does not bind to the insulin receptor (IR). This aspect indicates that the antibodies described herein may have adverse effects on IR, i.e., insulin metabolism. It attracts attention because it doesn't do that.

[0193] In another embodiment, yet another advantage of this antibody is that it is only effective against human IGF-1R. It can also bind to monkey IGF-1R, or more specifically, cynomolgus monkey IGF-1R. This aspect is also noteworthy because it helps in toxicity assessments required for clinical trials.

[0194] In yet another embodiment, the antibody consists of a monoclonal antibody. Clonal antibodies include mouse, chimeric, and humanized antibodies, as described below. It can be done.

[0195] Antibodies are preferably French collection for microor ganism cultures (CNCM, Pasteur Institute, 25 rue du Docteur Roux, 75724 Paris Cedex 15, France Derived from a submitted mouse hybridoma, the hybridoma is Balb / C By fusing immune mouse splenocytes / lymphocytes with myeloma Sp2 / O-Ag 14 cell line cells This is what was obtained.

[0196] In one embodiment, the IGF-1R antibody is derived from a mouse antibody, and therefore, m[antibody It is referred to as [name].

[0197] In one embodiment, the IGF-1R antibody is a chimeric antibody, and therefore the name of the antibody It is referred to as ].

[0198] In one embodiment, the IGF-1R antibody consists of a humanized antibody, and therefore, hz[antibody It is referred to as [the name of].

[0199] To avoid any doubt, the following specification will use the terms "IGF-1R antibody" and "[Antibody Name]". The expression "is equivalent to (unless otherwise specified) the IGF-1R antibody or the Includes mouse, chimeric, and humanized forms of "[Antibody Name]". If necessary, include the prefix m- (Mouse), c-(chimera), or hz-(humanized) are used.

[0200] For clarity, Table 2 below shows the definitions of preferred antibodies by IMGT. The CDR sequence is shown.

[0201] [Table 13]

[0202] Any combination of the six CDRs described above should be considered part of the present invention. This should be obvious to those skilled in the art.

[0203] As can be seen from Table 2, all antibodies described herein are CDR-H2, CDR -H3 and CDR-L2 have the same sequence, and this characteristic is particularly noteworthy as mentioned above. It is something that is done.

[0204] According to certain aspects, the antibody is derived from a species different from that of the mouse, particularly from a human antibody. This mouse antibody is characterized by including both light chain and heavy chain constant regions.

[0205] According to another specific aspect, the antibody is different from that of a mouse, and in particular, a human antibody. This is a chimeric (c) antibody characterized by including the derived light chain and heavy chain constant regions. .

[0206] A chimeric antibody is formed by combining the naturally variable regions (light chain and heavy chain) derived from an antibody of a given species with the given The term "species" refers to a combination of the constant regions of the light and heavy chains of heterologous antibodies.

[0207] Chimeric antibodies can be produced using recombinant genetics techniques. For example, Chimeric antibodies are promoters and non-human monoclonal antibodies, particularly variable mice. A sequence encoding a region, and a heterologous sequence, preferably a sequence encoding a human antibody constant region. It can be produced by cloning recombinant DNA containing one such recombinant DNA. The chimeric antibody of the ADC according to the present invention, which is encoded by a gene, is, for example, mouse-human chimeric antibody. It may be Mera, and the specificity of this antibody is determined by a variable region derived from mouse DNA. Its isotype is determined by a constant region derived from human DNA.

[0208] According to one embodiment of the present invention, the antibody is a) Three heavy chain CDRs of sequences 7, 2 and 3, as well as sequences 9, 5 and An antibody comprising three light chain CDRs of sequence 11; b) Three heavy chain CDRs of sequences 7, 2 and 3, and sequences 10 and 5 An antibody comprising three light chain CDRs of sequence 11; c) Three heavy chain CDRs of sequences 7, 2 and 3, as well as sequences 9, 5 and An antibody comprising three light chain CDRs of sequence 12; and d) Three heavy chain CDRs of sequences 8, 2 and 3, as well as sequences 9, 5 and An antibody comprising three light chain CDRs of sequence 11. Selected from.

[0209] In a preferred embodiment, the antibody is, a) The sequence of sequence number 13 or any sequence that shows at least 80% identity with sequence number 13. The heavy chain variable domain of the sequence, as well as the three light chain CDRs of sequences 9, 5, and 11. An antibody comprising the components of; b) The sequence of sequence number 14 or any sequence that shows at least 80% identity with sequence number 14. The heavy chain variable domain of the sequence, as well as the three light chain CDs of sequences 10, 5, and 11. An antibody containing R; c) The sequence of sequence number 15 or any sequence that shows at least 80% identity with sequence number 15. The heavy chain variable domain of the sequence, as well as the three light chain CDRs of sequences 9, 5, and 12. An antibody comprising the components of; d) The sequence of sequence number 16 or any sequence that shows at least 80% identity with sequence number 16. The heavy chain variable domain of the sequence, as well as the three light chain CDRs of sequences 9, 5, and 11. Antibodies containing the following; and e) The sequence of sequence number 17 or any sequence that shows at least 80% identity with sequence number 17. The heavy chain variable domain of the sequence, as well as the three light chain CDRs of sequences 9, 5, and 12. Antibodies containing Selected from.

[0210] "Sequences 13-17 and at least 80%, preferably 85%, 90%, 95% "Any sequence showing 98% identity" refers to three heavy chain CDRs numbered 1, 2, and 3. In addition, sequences other than those corresponding to CDR (i.e., sequence numbers 1, 2, and 3) All sequence numbers 13-17 and at least 80%, preferably 85%, 90%, and 95% It is intended to represent sequences that exhibit 98% identity.

[0211] According to one embodiment of the present invention, the antibody is a) Heavy chain variable domain of sequence number 13, and sequence numbers 9, 5 and 11 An antibody comprising three light chain CDRs in a column; b) The heavy chain variable domain of sequence number 14, and sequence numbers 10, 5 and 11 An antibody comprising three light chain CDRs of a sequence; c) Heavy chain variable domain of sequence number 15, and sequence numbers 9, 5 and 12 An antibody comprising three light chain CDRs in a column; d) The heavy chain variable domain of sequence number 16, and the sequences of sequence numbers 9, 5 and 11 An antibody comprising three light chain CDRs in a row; and e) Heavy chain variable domain of sequence number 17, and sequence numbers 9, 5 and 12 An antibody comprising three light chain CDRs in a row. Selected from.

[0212] In another preferred embodiment, the antibody is, a) The sequence of sequence number 18 or any sequence that shows at least 80% identity with sequence number 18. The light chain variable domain of the sequence, as well as the three heavy chain CDRs of sequences 7, 2, and 3 Antibodies containing; b) The sequence of sequence number 19 or any sequence that shows at least 80% identity with sequence number 19. The light chain variable domain of the sequence, as well as the three heavy chain CDRs of sequences 7, 2, and 3 Antibodies containing; c) The sequence of sequence number 20 or any sequence that shows at least 80% identity with sequence number 20. The light chain variable domain of the sequence, as well as the three heavy chain CDRs of sequences 7, 2, and 3 Antibodies containing; d) The sequence of sequence number 21 or any sequence that shows at least 80% identity with sequence number 21. The light chain variable domain of the sequence, as well as the three heavy chain CDRs of sequences 8, 2, and 3 Antibodies containing; and e) The sequence of sequence number 22 or any sequence that shows at least 80% identity with sequence number 22. The light chain variable domain of the sequence, as well as the three heavy chain CDRs of sequences 7, 2, and 3 Antibodies containing Selected from.

[0213] "Sequences 18-22 and at least 80%, preferably 85%, 90%, 95% and "Any sequence showing 98% identity" refers to the three light chain CDRs of sequence numbers 4, 5, and 6. In addition, it shows sequences other than those corresponding to CDR (i.e., sequences 4, 5, and 6). The full-length sequence of sequence numbers 18-22 and at least 80%, preferably 85%, 90%, 95% These are intended to represent sequences indicating % and 98% identity, respectively.

[0214] According to one embodiment of the present invention, the antibody is a) The light chain variable domain of sequence number 18, and the sequences of sequence numbers 7, 2, and 3. An antibody comprising three heavy chain CDRs; b) The light chain variable domain of sequence number 19, and the sequences of sequence numbers 7, 2, and 3. An antibody comprising three heavy chain CDRs; c) The light chain variable domain of the sequence of sequence number 20, and the sequences of sequence numbers 7, 2, and 3. An antibody comprising three heavy chain CDRs; d) The light chain variable domain of the sequence of sequence number 21, and the sequences of sequence numbers 8, 2, and 3 An antibody comprising three heavy chain CDRs; and e) The light chain variable domain of sequence 22, and the sequences of sequences 7, 2, and 3. An antibody comprising three heavy chain CDRs. Selected from.

[0215] According to one embodiment of the present invention, the antibody is a) The sequence of sequence number 13 or any sequence that shows at least 80% identity with sequence number 13. The heavy chain variable domain of the sequence, and the sequence of sequence number 18 or sequence number 18 and at least 8 An antibody comprising a light chain variable domain of any sequence exhibiting 0% identity; b) The sequence of sequence number 14 or any sequence that shows at least 80% identity with sequence number 14. The heavy chain variable domain of the sequence, and the sequence of sequence number 19 or sequence number 19 and at least 8 An antibody comprising a light chain variable domain of any sequence exhibiting 0% identity; c) The sequence of sequence number 15 or any sequence that shows at least 80% identity with sequence number 15. The heavy chain variable domain of the sequence, and the sequence of sequence number 20 or sequence number 20 and at least 8 An antibody comprising a light chain variable domain of any sequence exhibiting 0% identity; d) The sequence of sequence number 16 or any sequence that shows at least 80% identity with sequence number 16. The heavy chain variable domain of the sequence, and the sequence of sequence number 21 or sequence number 21 and at least 8 An antibody comprising a light chain variable domain of any sequence exhibiting 0% identity; and e) The sequence of sequence number 17 or any sequence that shows at least 80% identity with sequence number 17. The heavy chain variable domain of the sequence, and the sequence of sequence number 22 or sequence number 22 and at least 8 An antibody comprising a light chain variable domain of any sequence exhibiting 0% identity. It is an antibody selected from among them.

[0216] The chimeric antibodies described herein can also be characterized by a constant domain, More specifically, the aforementioned chimeric antibodies are not limited to IgG1 and IgG2. IgG3, IgM, IgA, IgD, or IgE can be selected and designed. More preferably, in relation to the present invention, the chimeric antibody is IgG1 or IgG4.

[0217] According to one embodiment of the present invention, the antibody is variable in IgG1 form as described above. A chimeric antibody comprising domains VH and VL. More preferably, the chimeric antibody The body has a constant domain of VH in sequence 43 and a κ in VL in sequence 45. It includes a domain.

[0218] According to one embodiment of the present invention, the antibody is variable in IgG4 form as described above. A chimeric antibody comprising domains VH and VL. More preferably, the chimeric antibody The body has a constant domain of VH in sequence 44 and a κ in VL in sequence 45. It includes a domain.

[0219] In another preferred embodiment, the antibody is, a) The sequence of sequence number 23 or any sequence that shows at least 80% identity with sequence number 23. The heavy chain of the sequence, and the sequence of sequence number 28 or at least 80% identity with sequence number 28. An antibody comprising or consisting of a light chain of any sequence exhibiting the following characteristics; b) The sequence of sequence number 24 or any sequence that shows at least 80% identity with sequence number 24. The heavy chain of the sequence, and the sequence of sequence number 29 or at least 80% identity with sequence number 29. An antibody comprising or consisting of a light chain of any sequence exhibiting the following characteristics; c) The sequence of sequence number 25 or any sequence that shows at least 80% identity with sequence number 25. The heavy chain of the sequence, and the sequence of sequence number 30 or at least 80% identity with sequence number 30. An antibody comprising or consisting of a light chain of any sequence exhibiting the following characteristics; d) The sequence of sequence number 26 or any sequence that shows at least 80% identity with sequence number 26. The heavy chain of the sequence, and the sequence of sequence number 31 or at least 80% identity with sequence number 31. Antibodies comprising or consisting of light chains of any sequence exhibiting; and e) The sequence of sequence number 27 or any sequence that shows at least 80% identity with sequence number 27. The heavy chain of the sequence, and the sequence of sequence number 32 or at least 80% identity with sequence number 32. An antibody comprising or consisting of a light chain of any sequence that exhibits the following characteristics Selected from.

[0220] To clarify further, Table 3 below shows the VH and VL values ​​for preferred chimeric antibodies. This shows the array.

[0221] [Table 14]

[0222] According to another specific aspect of the present invention, the antibody is derived from a human antibody and has a constant region of the light chain and The heavy chain is characterized by having either a λ or κ region and a γ-1, γ-2, or γ-4 region, respectively. This is a humanized antibody.

[0223] Humanized antibodies or their fragments can be produced by techniques known to those skilled in the art. Such humanized antibodies can be used for in vitro diagnosis or in vivo prediction. They are preferable for use in methods including preventive and / or therapeutic measures. For example, Patents EP0451216, EP0682040, EP0939127, EP by PDL 0566647 or US5,530,101, US6,180,370, US5,58 "CDR Graffiti" as described in 5,089 and US5,693,761 Other humanization technologies, such as the "G" technology, are also known to those skilled in the art. U.S. Patent No. 5,639,641 , Nos. 6,054,297, 5,886,152 and 5,877,293 The issue number can also be cited.

[0224] In a preferred embodiment, the antibody is i) CDR-H1, CDR-H2 and CD with sequences 7, 2, and 3 respectively R-H3, and ii) Human germline IGHV1-46 * FR1, F derived from 01 (sequence number 46) R2 and FR3, as well as iii) Human germline IGHJ4 * FR4 derived from 01 (Sequence ID 48) It comprises a heavy chain variable domain (VH) having the following characteristics.

[0225] In a preferred embodiment, the antibody is i) CDR-L1, CDR-L2, and CDR-L2 sequences of sequence numbers 9, 5, and 11, respectively. DR-L3, and ii) Human germline IGKV1-39 * FR1, F derived from 01 (sequence number 47) R2 and FR3, as well as iii) Human germline IGKJ4 * FR4 derived from 01 (Sequence ID 49) It comprises a light chain variable domain (VL) having the following characteristics.

[0226] In a preferred embodiment of the present invention, the antibody is, a) CDR-H1, CDR-H2 and CD with sequences 7, 2, and 3 respectively R-H3, and human germline IGHV1-46 * Derived from 01 (Sequence ID 46) FR1, FR2, and FR3, as well as human germline IGHJ4 * 01 (Sequence ID 48) ) heavy chain having FR4 derived from; and b) CDR-L1, CDR-L2 and CDR-L2 of sequence numbers 9, 5 and 11, respectively. DR-L3, and human germline IGKV1-39 * Derived from 01 (sequence number 47) FR1, FR2 and FR3, as well as human germline IGKJ4 * 01 (Sequence ID 4) 9) Light chain having FR4 derived from It consists of including.

[0227] In one embodiment, the antibody has a heavy chain variable domain (VH) of the sequence of SEQ ID NO: 33, The humanized antibody comprises the light chain variable domain (VL) of sequence number 35. Hereafter, this will be referred to as hz208F2 ("mutant" or "Var.1").

[0228] In another embodiment, the antibody has the sequence of SEQ ID NO: 33 as residues 20, 34, 35, 38, 48, 50, 59, 61, 62, 70, 72, 74, 76, 77, 79, 82 and 95 The sequence of sequence number 33, which includes at least one revert mutation selected from It comprises a heavy chain variable domain (VH).

[0229] In another embodiment, the antibody has the sequence of SEQ ID NO: 33 as residues 20, 34, 35, 38, 48, 50, 59, 61, 62, 70, 72, 74, 76, 77, 79, 82 and 95 Select from 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 , the heavy chain variable of the sequence of SEQ ID NO: 33, which includes 16 or 17 revert mutations. It includes the main (VH) track.

[0230] To clarify further, Table 4 below shows the preferred revertant mutations.

[0231] [Table 15]

[0232] In one embodiment, the antibody has a sequence at column number 35 consisting of residues 22, 53, 55, 65, and 7 A field comprising at least one revert mutation selected from 1, 72, 77, and 87 The combination comprises a light chain variable domain (VL) of the sequence of sequence number 35.

[0233] In one embodiment, the antibody has a sequence of residues 22, 53, 55, 65 of SEQ ID NO: 35 Sudden return of 2, 3, 4, 5, 6, 7, or 8, selected from 71, 72, 77, or 87 The sequence of sequence number 35, including the light chain variable domain (VL) when it contains a mutation .

[0234] In another embodiment, the antibody is a) The sequence of sequence number 33 consists of residues 20, 34, 35, 38, 48, 50, 59, 61, 6 At least one selected from 2, 70, 72, 74, 76, 77, 79, 82, and 95 The heavy chain variable domain (VH) of the sequence of sequence number 33 when it contains two revertant mutations and; b) The sequence of SEQ ID NO: 35 contains residues 22, 53, 55, 65, 71, 72, 77 and 87 The sequence of sequence number 35, which includes at least one revert mutation selected from Light chain variable domain (VL) It consists of including.

[0235] To clarify further, Table 5 below shows the preferred revertant mutations.

[0236] [Table 16]

[0237] In such embodiments, the antibody contains all of the above-mentioned reverse mutations, and sequence number The heavy chain variable domain (VH) of sequence 34, and the light chain variable domain of sequence 36 This corresponds to an antibody containing n(VL). The humanized antibody is hereafter referred to as hz208F2(" It is called "Variant 3" or "Var.3".

[0238] In another embodiment, all humanized forms included between mutant 1 and mutant 3 are also this This is encompassed by the invention. In other words, this antibody is included in the “consensus” formulation of SEQ ID NO: 41. The heavy chain variable domain (VH) of the column, and the light chain variable of the “consensus” sequence of sequence number 42. This corresponds to an antibody containing a domain (VL). The humanized antibody as a whole is as follows: This is called hz208F2 ("Variant 2" or "Var.2").

[0239] In a preferred embodiment, the antibody is, a) Sequence of sequence number 33 or sequence number 33 and at least 80%, preferably 85%, Heavy chain variable domains of any sequence exhibiting 90%, 95%, and 98% identity, as well as An antibody comprising three light chain CDRs of sequences 9, 5, and 11; and b) Sequence of sequence number 34 or sequence number 34 and at least 80%, preferably 85%, Heavy chain variable domains of any sequence exhibiting 90%, 95%, and 98% identity, as well as An antibody comprising three light chain CDRs with sequences at column numbers 9, 5, and 11. Selected from.

[0240] "Sequence number 33 or 34 and at least 80%, preferably 85%, 90%, 95% "Any sequence showing 98% identity" refers to the three heavy-chain CD sequences of sequence numbers 1, 2, and 3. It shows R, and in addition, the sequence outside of the sequence corresponding to CDR (i.e., sequence numbers 1, 2, and 3) The entire sequence of column number 33 or 34 and at least 80%, preferably 85%, 90%, or 95% And it is intended to represent sequences that show 98% identity.

[0241] In one embodiment of the present invention, the antibody is a) The sequence of sequence number 33 or any sequence that shows at least 80% identity with sequence number 33. The heavy chain variable domain of the sequence, as well as the three light chain CDRs of sequences 9, 5, and 11. An antibody comprising; and b) The sequence of sequence number 34 or any sequence that shows at least 80% identity with sequence number 34. The heavy chain variable domain of the sequence, as well as the three light chain CDRs of sequences 9, 5, and 11. Antibodies containing Selected from.

[0242] Unless otherwise indicated in the relevant paragraphs herein, any sequence is not a specific sequence. A sequence that shows at least 80% identity is one in which the sequence is at least 80% identical to the reference sequence. It must be understood that this indicates 85%, 90%, 95%, and 98% identity. i. If these sequences include the CDR sequence, those sequences are identical to the reference sequence CDR. At least these CDRs, and the remaining sequences located outside the sequences corresponding to these CDRs, The entire array and 80%, preferably 85%, 90%, 95% must be calculated. This is intended to indicate a 98% degree of identity.

[0243] In a preferred embodiment, the antibody is, a) Light chain variable domain of sequence number 35 or sequence number 35 and at least 80% Preferably any sequence exhibiting 85%, 90%, 95%, and 98% identity, as well as An antibody comprising three heavy chain CDRs of sequences 7, 2, and 3; and b) Sequence of sequence number 36 or sequence number 36 and at least 80%, preferably 85%, Light chain variable domains of any sequence exhibiting 90%, 95%, and 98% identity, as well as An antibody comprising three heavy chain CDRs with sequences 7, 2, and 3 in columns. Selected from.

[0244] "Sequence number 35 or 36 and at least 80%, preferably 85%, 90%, 95% "Any sequence showing 98% identity" refers to the three light chain CDs of sequence numbers 4, 5, and 6. R is shown, and in addition, sequences corresponding to CDR (i.e., sequence numbers 4, 5, and 6) The entire sequence outside sequence number 35 or 36 and at least 80%, preferably 85%, 90%, 9 It is intended to represent sequences exhibiting 5% and 98% identity.

[0245] In one embodiment of the present invention, the antibody is a) The sequence of sequence number 35 or any sequence that shows at least 80% identity with sequence number 35. The light chain variable domain of the sequence, as well as the three heavy chain CDRs of sequences 7, 2, and 3 Antibodies containing; and b) The sequence of sequence number 36 or any sequence that shows at least 80% identity with sequence number 36. The heavy chain variable domain of the sequence, as well as the three heavy chain CDRs of sequences 7, 2, and 3 Antibodies containing Selected from.

[0246] The humanized antibodies described herein can also be characterized by a constant domain, More specifically, the humanized antibodies mentioned above are not limited to IgG1, IgG2, and Ig G3, IgM, IgA, IgD, or IgE can be selected and designed. More specifically, in relation to the present invention, the humanized antibody is IgG1 or IgG4.

[0247] According to one embodiment of the present invention, the antibody is variable in IgG1 form as described above. A humanized antibody comprising domains VH and VL. More preferably, the humanized antibody The body has the constant domain of VH in sequence 43 and the κ of VL in sequence 45. It includes a domain.

[0248] According to one embodiment of the present invention, the antibody is variable in IgG4 form as described above. A humanized antibody comprising domains VH and VL. More preferably, the humanized antibody The body contains the constant domain of VH in sequence 44 and the κ domain of VL in sequence 45. It includes the main dish.

[0249] According to yet another embodiment of the present invention, the antibody is a) The sequence of sequence number 37 or any sequence that shows at least 80% identity with sequence number 37. The heavy chain of the sequence, and the sequence of sequence number 39 or at least 80% identity with sequence number 39. Antibodies comprising or consisting of light chains of any sequence exhibiting; and b) The sequence of sequence number 38 or any sequence that shows at least 80% identity with sequence number 38. The heavy chain of the sequence, and the sequence of sequence number 40 or at least 80% identity with sequence number 40. An antibody comprising or consisting of a light chain of any sequence that exhibits the following characteristics Selected from.

[0250] To make it clearer, Table 6a below shows variant 1 (Var.) of the humanized antibody hz208F2. 1) and an unspecified example of the VH and VL sequences of variant 3 (Var.3) is shown. This also includes the consensus sequence for variant 2 (Var.2).

[0251] [Table 17]

[0252] In another preferred embodiment, the antibody is, a) Sequence IDs 56, 62, 64, 66, 68, 70, 72, 74, 76, 78 and 8 Arrays or arrays selected from 0: 56, 62, 64, 66, 68, 70, 72, 74 , 76, 78 and 80 and at least 80%, preferably 85%, 90%, 95% and Heavy chain variable domains of any sequence with 98% identity; as well as Sequence IDs 9, 5 and An antibody comprising three light chain CDRs of sequence 11; b) An array selected from sequence number 57 or 60, or sequence number 57 or 60 and A unit having at least 80%, preferably 85%, 90%, 95%, and 98% identity. Light chain variable domain of sequence 7, 2, and 3; and three heavy chain CDs of sequence 7, 2, and 3. An antibody containing R; and c) Sequences 56, 62, 64, 66, 68, 70, 72, 74, 76, 78 and 8 Arrays or arrays selected from 0: 56, 62, 64, 66, 68, 70, 72, 74 , 76, 78 and 80 and at least 80%, preferably 85%, 90%, 95% and A heavy chain variable domain of any sequence with 98% identity; and Sequence ID No. 57 or At least 80% of the sequence selected from 60 or sequence number 57 or 60, preferably The light chain variable domain of any sequence having 85%, 90%, 95%, and 98% identity. Antibodies containing Selected from.

[0253] According to yet another embodiment of the present invention, the antibody is a) Sequence IDs 56, 62, 64, 66, 68, 70, 72, 74, 76, 78 and 8 Array of 0 or arrays 56, 62, 64, 66, 68, 70, 72, 74, 76, 78 Alternatively, a heavy chain of any sequence showing at least 80% identity with 80, and Sequence ID No. 5 The light chain contains any sequence that exhibits at least 80% identity with sequence number 7 or sequence number 57. Antibodies that are made of; and b) Sequences of sequence numbers 56, 64, 68 and 78 or sequence numbers 56, 64, 68 if k is a heavy chain of any sequence that shows at least 80% identity with 78, and sequence number 60 The sequence or any sequence light chain exhibiting at least 80% identity with sequence number 60 is not included. antibodies Selected from.

[0254] According to yet another embodiment of the present invention, the antibody is a) The sequence of sequence number 58 or any sequence that shows at least 80% identity with sequence number 58. The heavy chain of the sequence, and the sequence of sequence number 59, or at least 80% identical to sequence number 59. An antibody comprising or consisting of a light chain of any sequence exhibiting sex; b) The sequence of sequence number 58 or any sequence that shows at least 80% identity with sequence number 58. The heavy chain of the sequence, and the sequence of sequence number 61 or at least 80% identity with sequence number 61. An antibody comprising or consisting of a light chain of any sequence exhibiting the following characteristics; c) The sequence of sequence number 63 or any sequence that shows at least 80% identity with sequence number 63. The heavy chain of the sequence, and the sequence of sequence number 59 or at least 80% identity with sequence number 59. An antibody comprising or consisting of a light chain of any sequence exhibiting the following characteristics; d) The sequence of sequence number 65 or any sequence that shows at least 80% identity with sequence number 65. The heavy chain of the sequence, and the sequence of sequence number 59 or at least 80% identity with sequence number 59. An antibody comprising or consisting of a light chain of any sequence exhibiting the following characteristics; e) The sequence of sequence number 65 or any sequence that shows at least 80% identity with sequence number 65. The heavy chain of the sequence, and the sequence of sequence number 61 or at least 80% identity with sequence number 61. An antibody comprising or consisting of a light chain of any sequence exhibiting the following characteristics; f) The sequence of sequence number 67 or any sequence that shows at least 80% identity with sequence number 67. The heavy chain of the sequence, and the sequence of sequence number 59 or at least 80% identity with sequence number 59. An antibody comprising or consisting of a light chain of any sequence exhibiting the following characteristics; g) The sequence of sequence number 69 or any sequence that shows at least 80% identity with sequence number 69. The heavy chain of the sequence, and the sequence of sequence number 59 or at least 80% identity with sequence number 59. An antibody comprising or consisting of any sequence light chain exhibiting the following characteristics; h) The sequence of sequence number 69 or any sequence that shows at least 80% identity with sequence number 69. The heavy chain of the sequence, and the sequence of sequence number 61 or at least 80% identity with sequence number 61. An antibody comprising or consisting of a light chain of any sequence exhibiting the following characteristics; i) The sequence of sequence number 71 or any sequence that shows at least 80% identity with sequence number 71 The heavy chain of the sequence, and the sequence of sequence number 59 or at least 80% identity with sequence number 59. An antibody comprising or consisting of a light chain of any sequence exhibiting the following characteristics; j) The sequence of sequence number 73 or any sequence that shows at least 80% identity with sequence number 73. The heavy chain of the sequence, and the sequence of sequence number 59 or at least 80% identity with sequence number 59. An antibody comprising or consisting of a chain light of any sequence exhibiting the following characteristics; k) The sequence of sequence number 75 or any sequence that shows at least 80% identity with sequence number 75. The heavy chain of the sequence, and the sequence of sequence number 59 or at least 80% identity with sequence number 59. An antibody comprising or consisting of a light chain of any sequence exhibiting the following characteristics; l) The sequence of sequence number 77 or any sequence that shows at least 80% identity with sequence number 77. The heavy chain of the sequence, and the light chain of the sequence of sequence number 59, or sequence number 59 and at least 80% An antibody comprising or consisting of any sequence that indicates identity; m) An array of sequence number 79 or any array that shows at least 80% identity with sequence number 79. The heavy chain of the sequence, and the sequence of sequence number 59 or at least 80% identity with sequence number 59. An antibody comprising or consisting of a light chain of any sequence exhibiting the following characteristics; n) The sequence of sequence number 79 or any sequence that shows at least 80% identity with sequence number 79. The heavy chain of the sequence, and the sequence of sequence number 61 or at least 80% identity with sequence number 61. Antibodies comprising or consisting of light chains of any sequence exhibiting; and o) The sequence of sequence number 81 or any sequence that shows at least 80% identity with sequence number 81. The heavy chain of the sequence, the sequence of sequence number 59, or shows at least 80% identity with sequence number 59. Antibodies comprising or consisting of light chains of any sequence. Selected from.

[0255] In other words, antibodies are a) Sequence IDs 58, 63, 65, 67, 69, 71, 73, 75, 77, 79 and 8 Arrays selected from 1 or array numbers 58, 63, 65, 67, 69, 71, 73, 75 , heavy chains of any sequence having at least 80% identity with 77, 79 and 81; and to b) Sequences selected from sequence numbers 59 and 61 or sequence numbers 59 and 61 and fewer A light chain of any sequence having at least 80% identity It may be an antibody that contains [a certain substance].

[0256] In one embodiment of the present invention, the antibody is a) Sequence IDs 58, 63, 65, 67, 69, 71, 73, 75, 77, 79 and 8 Arrays selected from 1 or array numbers 58, 63, 65, 67, 69, 71, 73, 75 , a heavy chain of any sequence having at least 80% identity with 77, 79 or 81; Bini b) Sequences selected from sequence numbers 59 and 61, or sequences from sequence number 59 or 61 and a smaller number of sequences. A light chain of any sequence that has at least 80% identity. Selected from.

[0257] To clarify further, Table 6b below shows the various variants of the humanized antibody hz208F2. The following are unrestricted examples of VH and VL (variable domain and full-length) sequences.

[0258] [Table 18]

[0259] According to another aspect of the present invention, the antibody is i) May 30, 2013 and 2013 respectively June 26, July 26, 2013, April 24, 2013, and July 26, 2013 Hybridoma I-4757, deposited at the CNCM (Pasteur Institute, France), Antibodies produced by I-4773, I-4775, I-4736, or I-4774 or ii) an antibody that competes with the antibody in i) for binding to IGF-1R; or iii) Select an antibody that binds to the same epitope as the antibody in i) of IGF-1R. It is an antibody that can be released.

[0260] According to certain aspects, the binding unit is a means of delivering cytotoxic drugs to host target sites. Antibodies, or their antigen-binding agents, as described above, for use as dressing vehicles. The lagment is characterized by the host target site being IGF-1R, preferably IGF-1R cells. External domain, more preferably human IGF-1R (SEQ ID NO: 50) and even more preferably More preferably, the human IGF-1R extracellular domain (SEQ ID NO: 51), and even more preferably. , the N-terminus of the human IGF-1R extracellular domain (SEQ ID NO: 52), or any intrinsic variant thereof. It consists of epitopes localized in heteromorphic arrangements.

[0261] In a preferred embodiment, the host target site is a mammalian cell, more preferably a hi IGF-1R expresses in cells, more preferably naturally or by genetic engineering. It is the target site of the cell.

[0262] In further embodiments, the host target site is cancer, preferably IGF-1R expression Patients with cancer or IGF-1R-related cancer, preferably target sites of human cells .

[0263] IGF-1R-expressing cancers or IGF-1R-related cancers are characterized in particular by tumor cells that are complete on their surface. This includes cancers that express or overexpress all or part of IGF-1R.

[0264] In this invention, the IGF-1R antibody that can be used as a binding unit is particularly WO2015 / Listed in 162291, WO2015 / 162292 or WO2015 / 162293 It is being done.

[0265] Linker molecule Preferably, the linker of formula (I) according to the present invention has q=2, and the drug is transmitted to an antibody (for example, Covalently linked to binding units such as monoclonal antibodies or their antigen-binding fragments. It is useful for concluding a relationship.

[0266] Therefore, the sulfomaleimide portion of the linker is the thiol portion present on the bond unit. It can react with, while the X3 terminus of the linker is located on the drug (QH or Q-OH). It can react with the functional groups present.

[0267] Linker molecules can be prepared according to the various synthesis methods exemplified in the experimental section. .

[0268] If X1 and X2 are independently selected from H and Cl (at least one is Cl) The linker according to the present invention is of the formula L-NHCO-CH2CH2-SS-CH2CH2 -CONH-L(where L is L1-(CO) c -(W) w -(Y) y (Represents -X3) Depending on the circumstances, from protected forms of disulfide compounds to chlorinating agents such as SO2Cl2. It can be produced through a reaction with [the specified substance].

[0269] If X1 and X2 are independently selected from H and Br (at least one of them is Br) The linker according to the present invention is a form that is optionally protected, formula [ka] (Here, L is L1-(CO) c -(W) w -(Y) y (Represents -X3) These compounds, in some cases, are prepared by reaction with brominating agents such as Br2 from protected forms of the compound. It is possible.

[0270] At least one of X1 and X2 may be replaced depending on the circumstances (C1-C6). Alkoxy, aryloxy, or -O-(CH2CH2O) r When H, the present invention The linker is, in some cases, in a protected form, where at least one of X1 and X2 is C From the linker of the corresponding formula (I), which is l or Br, R a (C1-C6) alkyl, Aryl or -(CH2CH2O) which may be substituted in some cases. r Formula R representing H a It can be prepared by a nucleophilic substitution reaction of the -OH group with an alcohol.

[0271] Also, end-cut type linker portion (for example, L=L1-(CO) c -X6 has X6 official The active group is, for example, NH2, OH, or a leaving group, and may be in a protected form. (May also form a grafted sulfomaleimide moiety, and in particular, drug re Regarding the synthesis of the NKER complex, after the formation of the sulfomaleimide moiety as shown below, phosphorus It is expected that the car synthesis will be completed.

[0272] Furthermore, the oxidation process involves S(O) q The group is brought to the required oxidation state (i.e., preferably q = It is feasible to convert to 2). Such oxidation processes are well known to those skilled in the art. The oxidizing agent used could be, for example, mCPBA, RuO4, or RuCl3 / NaIO4. ru.

[0273] Further protection / deprotection processes can be carried out in the above process, and such processes The conditions for these reactions are well known to those skilled in the art.

[0274] The resulting linker can be obtained by extraction, evaporation or precipitation of the solvent or crystallization (and then filtration). It can be separated from the reaction medium by methods well known to those skilled in the art, such as [method name].

[0275] The linker can, if necessary, perform recrystallization, distillation, and chromatography on a silica gel column. Alternatively, it may be purified by methods well known to those skilled in the art, such as high-performance liquid chromatography (HPLC). It is also possible.

[0276] Drug linker complex Preferably, the drug linker conjugate of formula (II) according to the present invention, where q=2, is used to deliver the drug to the antibody ( For example, covalent binding units such as monoclonal antibodies or their antigen-binding fragments It is useful for linking things together.

[0277] Therefore, the sulfomaleimide portion of the drug linker complex has a thi present on the binding unit. It can react with the oars.

[0278] Drug linker complexes can be prepared according to various synthesis methods. In fact, formula ( I) The linker reacts with the drug (QH or Q-OH) to form a complex. This can be done. However, linkers are formed stepwise on the drug molecule, that is, phosphorus The first portion of Kerr is first grafted onto the drug, and the resulting compound is a terminally cleaved phosphorus. Another possibility to consider is that it reacts with Kerr molecules to form a drug-linker complex. can.

[0279] Therefore, although other synthesis routes may be considered, the following unlimited synthesis routes are according to the present invention. It can be used to prepare drug linker complexes of formula (II).

[0280] In all of these synthesis routes, further protection / deprotection / substitution steps can be performed. Such processes and their reaction conditions are well known to those skilled in the art.

[0281] The resulting drug linker complex is extracted, evaporated or precipitated or crystallized (then, It can be separated from the reaction medium by methods well known to those skilled in the art, such as by filtration.

[0282] Drug linker complexes can be recrystallized, distilled, and chromatographed on silica gel columns, if necessary. By methods well known to those skilled in the art, such as Raffie or high-performance liquid chromatography (HPLC) It can also be refined.

[0283] Synthesis path I represented by scheme I: [ka] The terminal sulfomaleimide portion is a precursor of the linker portion, as detailed below in the graph. It can be formed from a group already present in the drug being treated.

[0284] Step 1: 3-(2-chlorocarbonyl-ethyldisulfanyl)-propionyl chloride The formula is H2N-L1-(CO) c -(W) w -(Y) y -The molecule of Q (i.e., the linker) The part reacts with the drug molecule (which has already been grafted). Such a reaction is performed by trimeth This reaction can be carried out in the presence of a base such as ruamine. This reaction can be carried out in a solvent such as DCM, This can be done at temperatures ranging from 0°C to room temperature.

[0285] 3-(2-chlorocarbonyl-ethyldisulfanyl)-propionyl chloride is 3 From ,3'-dithiodipropionic acid, acyl chlorides are formed by reactions such as the reaction with (COCl)2. It can be prepared by well-known methods for forming it. This reaction is carried out by DCM and other similar methods. This can be carried out in a solvent, particularly at room temperature. A catalytic amount of DMF can be added.

[0286] Also, 3-(2-chlorocarbonyl-ethyldisulfanyl)-propionyl chloride For example, a protected form of the formula H2N-L1-(CO) c -X3 molecule and reaction In response, in subsequent steps, Q was grafted according to one of the other synthesis routes described below. It is also conceivable that the synthesis of the linker portion could be completed.

[0287] Step 2: The molecules obtained in Step 1, in particular, in large excess amounts (e.g., 5-10 equivalents, e.g., approximately It can be cyclized and chlorinated in the presence of SO2Cl2 (9 equivalents). The process can be carried out in a solvent such as DCM, especially at room temperature.

[0288] The chlorine atom is replaced by another X1 or X2 group (other than H) by known methods such as nucleophilic substitution. It can be converted to ).

[0289] Step 3: If necessary, oxidize the molecule obtained in Step 2 to obtain the drug-phosphorus of formula (IIa). Car is obtained. Such oxidation steps are carried out under conditions well known to those skilled in the art, in particular, with mCPBA (e.g. For example, it can be carried out in the presence of 10 equivalents. This reaction can be carried out in a solvent such as DCM, in particular It can be done at room temperature.

[0290] Synthesis Path II, represented by Scheme II: [ka] Direct coupling of a drug (QH or Q-OH) with the linker of formula (I) is possible. This is possible, and the conditions for this vary depending on the properties of X3 and the functional groups present on the drug.

[0291] This coupling can be a nucleophilic substitution or a substitution such as the Mitsunobu reaction, and such chemical reactions The reaction conditions for the response are well known to those skilled in the art.

[0292] X3 = OH and at least y = 1, w ≠ 0 or c = 1 (i.e., phosphorus in equation (I)) If the terminal functional group of Kerr is COOH and QH contains an NH functional group, then formula (I) The coupling between the linker and the drug (QH) is a peptide coupling known to those skilled in the art. It is possible. The terminal COOH functional group can also be converted to acyl COCl chloride, then This is reacted with a nucleophilic functional group (e.g., NH or OH) present on the drug (QH). It is possible.

[0293] X3=NH2; or X3=H, y=z=1 and Z=-NR4-(CH2) u -NR 5-; or X3=H, c=w=y=0, z'=1 and Z' is -NR4-(CH2) u - NR5- (that is, the terminal functional group of the linker in formula (I) is NH) and Q-OH If it contains a COOH functional group, the cup between the linker and the drug (Q-OH) in formula (I) The ring may be a peptide coupling known to those skilled in the art. Terminal COOH functionalization of the drug. The group can also be converted to acyl chloride (COCl), which is then reacted with an NH functional group. It is possible.

[0294] S(O) as needed q The group is changed to the desired oxidation state (i.e., preferably q=2). An additional oxidation process can be performed to convert it. Such oxidation processes are well known to those skilled in the art. Yes, there are. The oxidizing agents used are, for example, mCPBA, RuO4, or RuCl3 / NaIO4. It is possible.

[0295] Synthesis route III, represented by schemes IIIa and IIIb: [ka] Peptide coupling is performed as shown in schemes IIIa and IIIb, COOH Between the functional group-containing terminally cleaved linker and the grafted drug portion of the linker However, this can also be done. The terminal COOH functional group of a terminal cleavage linker is also acyl C chloride. It can also be converted to OCl, which can then be reacted with the NH functional group of other reactants. The reaction conditions for such a reaction are well known to those skilled in the art.

[0296] S(O) as needed q The group is changed to the desired oxidation state (i.e., preferably q=2). To replace, for example, in the presence of mCPBA, RuO4, or RuCl3 / NaIO4 An additional oxidation process can be performed.

[0297] Synthesis Path IV represented by Scheme IV: [ka] As shown in scheme IV, where X5 represents an OH group or a previously defined leaving group, The sulfomaleimide portion and the rest of the linker are already grafted into the drug. It can also perform coupling.

[0298] Coupling can be a type of substitution, such as a nucleophilic substitution.

[0299] S(O) as needed q The group is changed to the desired oxidation state (i.e., preferably q=2). To replace, for example, in the presence of mCPBA, RuO4, or RuCl3 / NaIO4 An additional oxidation process can be performed.

[0300] Synthesis path V represented by schemes Va and Vb: [ka] The linker consists of a heteroarylene moiety, which is a divalent 1H-1,2,3-tria If it is a zole, this heteroarylene group is L2 -(CH2) n -or-(CH 2CH2O) mIt represents -CH2-CH2- and L3 is -(CH2) p -or-(C H2CH2O) m -CH2-CH2- represents the base -(CH2CH 20) m As shown in the above schemes Va and Vb when it is not -CH2-CH2- formed by click chemistry between azide and alkyne under conditions well known to those skilled in the art. It is possible.

[0301] S(O) as needed q The group is changed to the desired oxidation state (i.e., preferably q=2). To replace, for example, in the presence of mCPBA, RuO4, or RuCl3 / NaIO4 An additional oxidation process can be performed.

[0302] Binding unit drug complex Antibody-drug conjugates and other binding unit drug conjugates are 1) In particular, the reduction of the disulfide bond forms a thiol functional group on the bond unit. and; and 2) The sulfomaleimide functional group reacts with the thiol functional group to form the drug moiety on the bond unit. The drug linker complex is formed by covalently bonding the bond unit having a thiol functional group to the drug linker complex. Make them respond It can be produced by [method].

[0303] This method is shown in Scheme VI below.

[0304] [ka]

[0305] Pharmaceutical composition The pharmaceutical composition according to the present invention comprises a binding unit drug complex of formula (III) or (IV) and a small amount of It comprises at least one pharmaceutically acceptable excipient.

[0306] The pharmaceutical composition of the present invention is administered enterally (e.g., orally) or parenterally (e.g., intravenously). Preferably, it may be intended for oral or intravenous administration. The active ingredient is mixed with conventional pharmaceutical excipients. Combined, unit dose form for administration to animals, preferably mammals including humans. ru.

[0307] For oral administration, the pharmaceutical composition is in solid or liquid form (solution or suspension). It is possible.

[0308] The solid composition may be in the form of tablets, gelatin capsules, powders, granules, etc. In this case, the active ingredients before compression are gelatin, starch, lactose, and magnesium stearate. It can be mixed with medicinal vehicles such as nesium, talc, and gum arabic. The tablets are In particular, it may be further coated with sucrose or other suitable materials, and it may have long-lasting activity. It may be treated to have delayed activity. In powder or granule formulations, the active ingredient is divided Mix with powders, wetting agents or anti-settling agents, and with deodorizers or sweeteners, or It can be granulated together. In gelatin capsules, the active ingredient can be hard or soft. The above-mentioned powder or granules, or the following, in a gelatin capsule. It can be introduced in the form of liquid compositions such as the following.

[0309] The liquid composition contains an active ingredient in a solvent such as water, along with a sweetener, flavor enhancer, or suitable coloring agent. They may both be contained. The liquid composition may also contain the aforementioned powders or granules with water, fruit juice, and milk. It can also be obtained by suspending or dissolving it in a liquid such as a chlorine. Liquid compositions include, for example, For example, it could be a syrup or an elixir.

[0310] For parenteral administration, the composition may contain an anti-precipitation agent and / or a wetting agent. It may be in the form of a suspension or a solution. This composition is advantageously sterile. This composition is, It may be in the form of a diuretic solution (especially for blood).

[0311] Such parenteral compositions are advantageously generally isotonic salt solutions, i.e., 0.9% NaC It contains a physiologically acceptable medium based on an aqueous solution (physiological saline). It also contains non-aqueous water. Miscible auxiliary solvents, such as ethanol, glycerin, propylene glycol or n- Lactamides can also be used.

[0312] The parenteral composition of the present invention also contains a precipitation inhibitor, a wetting agent, a preservative, an antioxidant, a chelating agent, It may also include one or more additives such as buffering agents and isotonic agents. Such additives are available to those skilled in the art. It is customary.

[0313] The anti-precipitation agents are alginate, sodium carboxymethylcellulose, and methylcellulose. hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose Ropil methylcellulose, microcrystalline cellulose, gum (e.g., gum arabic, tragacan) (Togum or xanthan gum), gelatin, carrageenan, polyvinylpyrrolidone, etc. could be.

[0314] The humectant is glycerin, propylene glycol, or a nonionic surfactant, for example, It may be lecithin, polysorbate, or poloxamer.

[0315] The preservatives are benzyl alcohol, phenol, cresol, chlorobutanol, and parabens. (For example, methylparaben, propylparaben or propylparaben), benzalkonium chloride These could be ruconium, benzethonium chloride, etc.

[0316] The antioxidants are ascorbic acid, citric acid, acetylcysteine, and sulfites (bisulfite). Salt, metabisulfite), monothioglycerol, sodium formaldehyde sulfoxylate These could be mu, thiourea, tocopherol, etc.

[0317] The chelating agent may be ethylenediaminetetraacetic acid (EDTA) salt.

[0318] The buffering agents are acetate, citrate, tartrate, phosphate, and triethanolamine (TRI S) is one possible example.

[0319] The isotonic agents are dextrose, glycerol, sodium chloride, glycerin, and mannitol. It could be a ru, etc.

[0320] The conjugated drug complex of the present invention contains a pharmaceutical composition containing 0.01 mg to 1000 mg per day. It can be used in a range of dosages, either once a day or several times a day, for example. It can be administered twice a day at the same dose. The daily dose is preferably 5 mg to 500 mg. Between g, it is more advantageous to include it between 10 mg and 200 mg. However, these ranges In some cases, it may be necessary to use a dosage outside of the standard range, which is something that a person skilled in the art would be aware of. ru.

[0321] cancer treatment A drug conjugate of formula (III) or (IV) or formula (III) or (I A pharmaceutical composition comprising the bonding unit of V) is particularly useful in the treatment of cancer, such as with cytotoxic drugs. If it contains a drug portion (Q) which is a residue of a useful drug (QH), it can be used to treat cancer. It is possible.

[0322] Antibody-drug conjugates (ADCs) and other binding unit drug conjugates are binding units such as antibodies. It possesses the combined effects of the opposite sex and, for example, the efficacy of cytotoxic drugs.

[0323] The use of conjugated drug complexes such as ADCs is normal when administered as a non-complexed drug. It enables local delivery of drugs that could cause unacceptable toxicity to cells. In other words, This therapy requires the greatest possible efficacy with minimal toxicity.

[0324] Cancer is not limited to this, but includes prostate cancer, osteosarcoma, lung cancer, breast cancer, endometrial cancer, and glioblastoma. Tumors, colon cancer, gastric cancer, kidney cancer, pancreatic cancer, head and neck cancer, or tumor cells targeted by antibodies This is exemplified by cancers associated with the expression of certain antigens.

[0325] The present invention is illustrated by the following examples and figures, which are not limited to this invention. [Brief explanation of the drawing]

[0326] [Figure 1A] Figure 1A shows the mass spectrum of the drug linker complex according to the present invention. [Figure 1B] Figure 1B shows the 1H-NMR spectrum of the drug linker complex according to the present invention. [Figure 2] Figure 2 shows the mass spectrum of the drug linker complex according to the present invention. [Figure 3A] Figure 3A shows the mass spectrum of the drug linker complex according to the present invention. [Figure 3B] Figure 3B shows the 1H-NMR spectrum of the drug linker complex according to the present invention. [Figure 4A] Figure 4A shows the mass spectrum of the drug linker complex according to the present invention. [Figure 4B] Figure 4B shows the 1H-NMR spectrum of the drug linker complex according to the present invention. [Figure 5A] Figure 5A shows the mass spectrum of the drug linker complex according to the present invention. [Figure 5B] Figure 5B shows the 1H-NMR spectrum of the drug linker complex according to the present invention. [Figure 6A] Figure 6A shows the mass spectrum of the drug linker complex according to the present invention. [Figure 6B] Figure 6B shows the 1H-NMR spectrum of the drug linker complex according to the present invention. [Figure 7A] Figure 7A shows the mass spectrum of the drug linker complex according to the present invention. [Figure 7B] Figure 7B shows the 1H-NMR spectrum of the drug linker complex according to the present invention. [Figure 8] Figure 8 shows the 1H-NMR spectrum of the drug linker complex according to the present invention. [Figure 9] Figure 9 shows the 1H-NMR spectrum of the drug linker complex according to the present invention. [Figure 10] Figure 10 shows the mass spectrum of the drug-somatostatin complex according to the present invention. [Figure 11] Figure 11 shows the mass spectrum of the drug-somatostatin complex according to the present invention. [Figure 12] Figure 12 shows the SDS-PAGE analysis of Ab1 antibody (1) and purified ADCs according to the present invention (ADC1-A (2), ADC1-B (3), ADC1-C (4), ADC1-D (5), ADC1-E (6), ADC1-F (7), and ADC1-G (8)) under reducing and non-reducing conditions. The bands observed on the gel correspond to fully crosslinked antibodies (i.e., LHHL); partially crosslinked antibodies (i.e., HHL, HH, HL); and non-crosslinked antibodies (i.e., H and L). [Figure 13]Figure 13 shows the SEC analysis of the Ab1 antibody and ADCs (ADC1-A, ADC1-B, ADC1-C, ADC1-D, ADC1-E, ADC1-F, and ADC1-G) according to the present invention. [Figure 14A] Figure 14A shows the ADC m / z spectrum of ADC:(A)ADC1-A according to the present invention before deconvolution. [Figure 14B] Figure 14B shows the ADC m / z spectrum of ADC(B)ADC1-B before deconvolution according to the present invention. [Figure 14C] Figure 14C shows the ADC m / z spectrum of ADC:(C)ADC1-C according to the present invention before deconvolution. [Figure 14D] Figure 14D shows the ADC m / z spectrum of the ADC:(D)ADC-1D according to the present invention before deconvolution. [Figure 15A] Figure 15A shows the DAR distribution after Maxent deconvolution of (A)ADC1-A. [Figure 15B] Figure 15B shows the DAR distribution after Maxent deconvolution of (B)ADC1-B. [Figure 15C] Figure 15C shows the DAR distribution after Maxent deconvolution of (C)ADC1-D. [Figure 16A] Figure 16A shows the analysis of ADC:(A) Reference ADC Ref-A by native mass spectrometry. [Figure 16B] Figure 16B shows the analysis of ADC1-C by native mass spectrometry according to the present invention. [Figure 17A] Figure 17A shows the results of in vitro stability tests for (A) reference ADC Ref-B, showing the percentage of total antibody (100%) and ADC in (1) human, (2) cynomolgus monkey, (3) mouse, and (4) rat serum at each time point (D0, D3, D7, and D14). [Figure 17B]Figure 17B shows the results of in vitro stability tests for (B)ADC1-C, showing the percentage of total antibody (100%) and ADC in (1) human, (2) cynomolgus monkey, (3) mouse, and (4) rat serum at each time point (D0, D3, D7, and D14). [Figure 17C] Figure 17C shows the results of in vitro stability tests for (C)ADC1-E, showing the percentage of total antibody (100%) and ADC in (1) human, (2) cynomolgus monkey, (3) mouse, and (4) rat serum at each time point (D0, D3, D7, and D14). [Figure 18A] Figure 18A shows the in vitro cytotoxicity evaluation of various ADCs in NCI-H2122 cells (A). [Figure 18B] Figure 18B shows the in vitro cytotoxicity evaluation of various ADCs in MCF-7 cells (B). [Figure 19] Figure 19 shows the in vivo activity of ADC1-C and reference ADC Ref-A in an ovarian cancer model. [Figure 20] Figure 20 shows the in vivo activity of ADC1-C and reference ADC Ref-A in an ovarian cancer model. [Figure 21] Figure 21 shows the mass spectrum of the drug linker complex according to the present invention. [Figure 22A] Figure 22A shows the mass spectrum of the drug linker complex according to the present invention. [Figure 22B] Figure 22B shows the TOF-MS spectrum of the drug linker complex according to the present invention. [Figure 23A] Figure 23A shows the SEC analysis of the ADC according to the present invention, synthesized using the Ab1 antibody and the PNU-159682 derivative. [Figure 23B] Figure 23B shows the SEC analysis of the ADC according to the present invention, synthesized using the Ab2 antibody and the PNU-159682 derivative. [Figure 24A]Figure 24A shows the ADC m / z spectrum of the ADC:(A)hz208F2-F562524 according to the present invention before deconvolution. [Figure 24B] Figure 24B shows the ADC m / z spectrum of the ADC:(B)c9G4-F562524 according to the present invention before deconvolution. [Figure 24C] Figure 24C shows the ADC m / z spectrum of the ADC:(C)hz208F2-F562616 according to the present invention before deconvolution. [Figure 24D] Figure 24D shows the ADC m / z spectrum of the ADC:(D)c9G4-F562646 according to the present invention before deconvolution. [Figure 25A] Figure 25A shows the DAR distribution after Maxent deconvolution of the ADC:(A)hz208F2-F562524 according to the present invention. [Figure 25B] Figure 25B shows the DAR distribution after Maxent deconvolution of the ADC:(B)c9G4-F562524 according to the present invention. [Figure 25C] Figure 25C shows the DAR distribution after Maxent deconvolution of the ADC:(C)hz208F2-F562616 according to the present invention. [Figure 25D] Figure 25D shows the DAR distribution after Maxent deconvolution of the ADC:(D)c9G4-F562646 according to the present invention. [Figure 26A] Figure 26A shows the in vitro evaluation of cytotoxicity of ADC hz208F2-F562524 and its corresponding control ADC c9G4-F562524 in NCI-H2122 cells (A). [Figure 26B] Figure 26B shows the in vitro cytotoxicity evaluation of ADC hz208F2-F562524 and its corresponding control ADC c9G4-F562524 in MCF-7 cells (B). [Figure 27A]Figure 27A shows the in vitro cytotoxicity evaluation of ADC hz208F2-F562646 and its corresponding control ADC c9G4-F562646 in NCI-H2122 cells (A). [Figure 27B] Figure 27B shows the in vitro cytotoxicity evaluation of ADC hz208F2-F562646 and its corresponding control ADC c9G4-F562646 in MCF-7 cells (B). [Figure 28] Figure 28 shows the in vivo activity of ADC hz208F2-F562524 and its corresponding control ADC c9G4-F562524 in an ovarian cancer model. [Examples]

[0327] Abbreviation

[0328] [Table 19]

[0329] Experimental procedure: All reactions requiring anhydrous conditions were carried out in oven-dried equipment under a nitrogen atmosphere. Anhydrous solvent The reagents were received in sealed bottles under an inert atmosphere. All reagents were used in the condition they were received. Column chromatography is performed using Interchim puriFlash® 4. 30 and Grace Reveleris® X2, silica gel (50μ m) The procedure was performed using a packed puriFlash® column. TLC was performed using silica (Merc). kSilica gel 60 F 254 This is done using aluminum sheets pre-coated with ) and UV-L Visualized at amp 254nm. Proton ( 1 H) and carbon ( 13 C) NMR Spectrometer Torrell is equipped with a BBO Prodigy probe (5mm) and a Bruker 500 M Recordings were made using Hz Ascend (trademark) at room temperature with CDCl3 and DMSO. The vector was interpreted using Topspin® 3.2 software. Chemical shift ( δ H and δ C ) is recorded at 1 part per million (ppm), and CDCl3( 1 H°NMR 7.26, 13 ¹¹C NMR 77.0 (center signal of the triplet) or DMSO ( 1 H NMR 2.50, 13 Refer to either C°NMR 37.9 or the central signal of the heptetate. The assignment will be to use the COSY experiment and the HSQC experiment as auxiliary. Coupling constant (J: adjacent Proton contact, J cis : The adjacent proton in the cis position, J o : Ortho proton, J m :meta The proton position is shown in Hertz up to the nearest ±0.1 Hz. The multiplicity is single, where appropriate. Line (s), double line (d), triple line (t), quadruple line (q), triple triple line (t of t) .), shown as multiline (m) and broad (b). The mass spectrum (m / z) is Wat Using the ers(registered trademark) ZQ Mass Detector spectrometer, electrospray Ionization method (ES+) technology, ion source temperature: 120°C, desolvation temperature: 350°C, capillary Lari voltage: 3.20kV, Cone voltage: 25V, Inlet electrode voltage: 5V, RF lens The data was recorded using a voltage of 0.5V and an MS scan range of 100-2000. HPLC analysis was then performed. This involves MassLynx 4.1 software and Waters 2996 PDA detection. HPLC with UV / vis (Waters® Alliance 269) In section 5, Waters(registered trademark) X-Bridge Shield RP18 3.5 μm (3.0 mm × 30 mm) column and Waters® X-Bridge Using a Shield RP18 3.5μm (3.0mm x 20mm) pre-column, The procedure was performed at a wavelength appropriate for the precipitated sample. Retention time (R t ) is the nearest 0.01 minutes This is shown. Two elution methods were used.

[0330] [Table 20]

[0331] [Table 21]

[0332] The retention time shown in Method 1 is R t,1 The holding time shown in Method 2 is R t,2 in show.

[0333] 1. Linker synthesis Example of an oxoisothiazolon synthesis pathway: [ka]

[0334] I.1. 3,3'-Disulfanediyldipropanoylchloride [ka] I.2. 3,3'-Disulfanediyldipropionic acid (4g, 0.019mol, 1 Suspend the equivalent amount in anhydrous DCM (100 mL), add anhydrous DMF (300 μL), and then the salt Add oxalyl oxalide (7.24 g, 0.057 mol, 3 equivalents) under an inert atmosphere at 0°C. The solution became clear. This mixture became clear and no gas was observed. It was left at room temperature for 3 hours until it reached this state. The crude product was evaporated and then maintained under reduced pressure for another 30 minutes. The residual oxalyl chloride was then removed. A yellow oily substance (4.70 g, 100%) was obtained. This crude product was used without further purification; R t,1 (MeOH): 2.13; M S ES+ m / z:206.86.

[0335] I.3. 6,6'-((3,3'-disulfanediylbis(propanoyl))bis( Dibenzyl dihexanoate (Azandiyl) [ka] 4-methylbenzenesulfonic acid 6-(benzyloxy)-6-oxohexane-1-a Minium (12.20 g, 0.031 mol, 2.2 equivalents) in an ice bath at 0°C, in an inert atmosphere. Under air pressure and vigorous stirring, the suspension was placed in anhydrous DCM (75 mL). TEA (1) was added to this solution. 5.72 mL, 0.113, 8 equivalents) were added. In a newly prepared 3DCM (25 mL) , 3'-disulfanediyldipropanoyl chloride (3.88g, 0.014mol, One equivalent of the solution was slowly added dropwise to a solution maintained at 0°C. Stirring was continued for 24 hours. The solution was allowed to reach room temperature during this time. Water (50 mL) was added, and this mixture was transferred to a separatory funnel. Organic layer The mixture is separated, washed with brine (1 × 100 mL), and then treated with 1 M HCl (1 × 100 mL). L), saturated NaHCO3 salt solution (2 × 100 mL) and brine again (1 × 100 mL) The layers were washed with ). The combined aqueous layers were extracted with DCM (2 x 100 mL). The organic layer was MgSO 4 is dried, filtered, and evaporated to dryness to obtain a yellow solid, which is then ground in MeOH and 6,6 '-((3,3'-disulfanjiilbis(propanoyl))bis(azanjiil)) Dibenzyl dihexanoate was obtained as a pale yellow powder (6.0 g, 66%). 1 1H NMR (500 MH) z, CDCl3), δ 7.35 (m, 10H), 6.0 (s, 2H), 5.11 (s, 4H), 3.25 (q, J=5.9 Hz, 4H), 3.00 (t, J=7.0 Hz, 4H), 2,55 (t, J=7.10 Hz, 4H), 2,.36 (t, J=7.30 Hz, 4H), 1.66 (t of t., J=8.10 Hz, 4H), 1.52 (t of t., J=8.10 Hz, 4H), 1.35 (t of t, J=8.52 Hz , 4H); 13 C NMR (500 MHz, CDCl3), δ 173.5 (2 -NH-C=O), 170.9 (-OC=O), 136.0 (aromatic 2C derived from the fragrant ring quat ), 128.6 (2 HC aromatic ), 128.3 (HC aromatic ), 128.2 (2 HC arom atic ), 66.2 (2 -CH2-O-), 39.4 (2 -CH2-N), 35.8 (2 -CH2-COO-), 34.3 (2 -CH2-S-), 34.1 (2 -CH2-CNO-), 29.1 (2 C), 26,3 (2 C), 24,4 (2 C); R t,1 (In MeOH): 2.50; MS ES+ M / Z: 617.00.

[0336] I.4. 6-(5-chloro-3-oxoisothiazole-2(3H)-yl)hexane benzyl acid [ka] 6,6'-((3,3'-disulfanediylbis(propanoyl))bis(azandi Dibenzyl dihexanoate (2.50g, 4.05mmol, 1 equivalent) is anhydrous DC It was dissolved in M ​​(20.3 mL). SO2Cl2 (97% purity, 2.96 mL) was added to this solution. Add 0.036 mol (9 equivalents) dropwise, and stir the mixture under an inert atmosphere at room temperature for 5 hours. Mixed. The solution became clear and pale yellow. Next, this solution was mixed with water (2 × 100 mL) Washed with ) and brine (1 x 100 mL). Combined aqueous layers were washed with DCM (2 x 100 mL). Extracted with L). The organic layer was dried over MgSO4, filtered, and then concentrated under reduced pressure. Purified using a chromatography column. 6-(5-chloro-3-oxoisothiazo Benzyl hexanoate (0.824g, 29.9%) is 6-(3- Oxoisothiazole-2(3H)-yl) together with benzyl hexanoate to form a pale yellow oily substance. I obtained it by doing so. 1 H NMR (CDCl3), δ 7.35 (m, 5H), 6.25 (s, 1H), 5.11 (s, 2H), 3.72 (t, J=7.34 Hz, 2H), 2.37 (t, J=7.39 Hz, 2H), 1.69 (m, 4H), 1.38 (m, 2H); 13 ¹ 0 MHz, CDCl3), δ 173.2 (-OC=O), 166.9 (-NC=O), 145.6 (Cl-HC=CH-), 136.0 (aromatic C derived from the group ring ruet. ), 128.6-128.3 (5 HC hromatic ), 114.8 (Cl-HC=CH-), 66.2 (-CH2-O- ), 43.5 (-CH2-N-), 34.0 (-CH2-C=O), 29.4, 25.9, 24.4; R t,1 (in ACN): 2.35; MS ES + m / z: 339.84.

[0337] I.5. 6-(5-chloro-1-oxide-3-oxoisothiazole-2(3H)- Benzyl hexanoate [ka] 6-(5-chloro-3-oxoisothiazole-2(3H)-yl)benzylhexanoate 3-chloroform (802g, 3.58 mmol) was diluted in anhydrous DCM (25mL). Add benzoperoxoic acid (1.2 equivalents). Allow this solution to steep at room temperature in an inert atmosphere for 48 hours. The mixture was stirred. Next, this solution was diluted with DCM and treated with a 10% Na2S2O3 aqueous solution. Next, the organic phase was treated with saturated NaHCO3 salt solution (2 × 100 mL), followed by brine (1 × Extraction was performed sequentially using 100 mL. The combined aqueous layer was extracted using DCM (2 × 100 mL). The chamber was dried with MgSO4, filtered, then concentrated under reduced pressure, and chromatographically analyzed. It was purified using rum. Then, 6-(5-chloro-1-oxide-3-oxoisothia) Benzyl zole-2(3H)-yl)hexanoate was obtained as a colorless oil (753 mg). ).

[0338] I.6. 6-(5-chloro-1,1-dioxide-3-oxoisothiazole-2(3 Benzyl hexanoate (H)-yl) [ka] Standard procedure for the oxidation of monochlorolide compounds: 6-(5-chloro-3-oxoisothiazole-2(3H)-yl)benzylhexanoate 1.11 g, 0.0036 mol, 1 equivalent of ru (amount) was diluted in anhydrous DCM (7 mL). Add 3-chlorobenzoperoxoic acid (2.69 g, 0.0109 mol, 3 equivalents). This solution was stirred at room temperature under an inert atmosphere for 48 hours. Next, this solution was diluted with DCM. Next, the organic phase was treated with a 10% Na2S2O3 aqueous solution. Then, the organic phase was treated with a saturated NaHCO3 salt solution (2 Extraction was carried out sequentially with (1 x 100 mL) and then with brine (1 x 100 mL). The combined aqueous layers were then... Extraction was performed using DCM (2 × 100 mL). The organic layer was dried over MgSO4, filtered, and then... The solution was concentrated under reduced pressure and purified using a chromatography column. 6-(5-chloro-1 ,1-dioxide-3-oxoisothiazole-2(3H)-yl)benzyl hexanoate It was obtained as a pale yellow oily substance (0.760 g, 64.3%). 1 1H NMR (CDCl3), δ 7.36 (m , 5H), 6.68 (s, 1H), 5.11 (s, 2H), 3.67 (t, J=7.68 Hz, 2H), 2.37 (t, J=7.27 Hz, 2H), 1.78 (t of t, J=7.73 Hz, 2H), 1.70 (t of t, J=7.38 Hz, 2H), 1.40 (m, 2H); 1 3 C NMR (500 MHz, CDCl3), δ 173.2 (-OC=O), 157.0 (-NC=O), 144.6 (Cl-HC=CH-), 1 36.0 (C derived from aromatic rings) oquet. ), 128.6-128.2 (HA romatic’s ), 123.6 (Cl-HC=CH-), 66 .2 (-CH2-O), 40.3 (-CH2-N-), 34.0 (-CH2-C=O), 27.9, 26.0, 24.2; R t,1 (in ACN): 2 .49; MS ES+ M / Z: 371.83.

[0339] Example 1. 6-(5-chloro-1,1-dioxide-3-oxoisothiazole-2( 3H)-yl)hexanoic acid [ka] 6-(5-chloro-1,1-dioxide-3-oxoisothiazole-2(3H)- Starting with benzyl hexanoate, and following the standard procedure for deprotection of benzyl esters, 6 -(5-chloro-1,1-dioxide-3-oxoisothiazole-2(3H)-yl) Hexanoic acid was obtained as a clearly colored powder (0.445 g, 77%). 1 1H NMR (CDCl3), δ 10. 89 (br. s, 1H), 6.70 (s, 1H), 3.70 (t, J=7.43 Hz, 2H), 2.38 (t, J=7.38 Hz, 2H), 1.81 (t of t., J=7.60 Hz, 2H), 1.69 (t of t, J=7.71 Hz, 2H), 1.43 (t of t division, J =7.75 Hz, J=3.31 Hz, 2H); 13 C NMR (500 MHz, CDCl3), δ 178.7 (O=C-OH), 157.1 (-N -C=O), 144.7 (Cl-HC=CH-), 123.6 (Cl-HC=CH-), 40.2 (-CH2-N), 33.5 (-CH2-C=O), 27. 9, 25.9, 23.9; R t,1 (in ACN): 1.98; MS ES+ m / z: 349.89.

[0340] I.7. 6-(4,5-dichloro-3-oxoisothiazole-2(3H)-yl) Benzyl xanate [ka] 6,6'-((3,3'-disulfanediylbis(propanoyl))bis(azandi Dibenzyl dihexanoate (3.21g, 0.0052mol, 1 equivalent) is anhydrous D It was dissolved in CM (26 mL). In this solution, SO2Cl2 (purity 97%, 3.80 mL) was added. Add 0.047 mol (9 equivalents) dropwise, and leave the mixture at room temperature under an inert atmosphere for 24 hours. The mixture was stirred. The solution became clear and pale yellow. Next, this mixture was added to water (2 × 100 mL) Washed with ) and brine (1 x 100 mL). Combined aqueous layers were washed with DCM (2 x 100 mL). Extracted with L). The organic layer was dried over MgSO4, filtered, and then concentrated under reduced pressure. Purified using a chromatography column. 6-(4,5-dichloro-3-oxoisothi Benzyl azole-2(3H)-yl)hexanoate was obtained as a pale yellow oily substance (1.39 1g, 35.7%). 1 H NMR (CDCl3), δ 7.35 (m, 5H), 5.11 (s, 2H), 3.79 (t, J=7. 18 Hz, 2H), 2.37 (t, J=7.33 Hz, 2H), 1.70 (m, 4H), 1.38 (m, 2H); 13 ¹¹¹ NMR (500 MHz) z, CDCl3), δ 173.2 (-OC=O), 161.9 (-NC=O), 138(-CH2-O-),.3 (Cl-CS-), 135.6 ( C derived from aromatic rings ruet ), 128.6-128.3 (5 Ha romatic ), 115.1 (Cl-CC=O), 66.2, 44.9 ( -CH2-N-), 33.9 (-CH2-C=O), 29.1, 25.9, 24.3; R t,1 (in ACN): 2.50; MS ES+ m / z: 37 3.75.

[0341] I.8. ​​6-(4,5-dichloro-1,1-dioxide-3-oxoisothiazole- Benzyl 2(3H)-yl)hexanoate [ka] Standard procedure for the oxidation of dichloride compounds: Ruthenium trichloride monohydrate (16 mg, 0.07 mmol, 0.013 equivalents) in water: In DCM:ACN (2:1:1, 1 ml), 6-(4,5-dichloro-3-oxoisothin Benzyl azole-2(3H)-yl)hexanoate (1.94g, 0.0052mol) It was added all at once to the stirred solution (1 equivalent). Next, sodium periodate (3.33 g, 0.0 Add 156 mol (3 equivalents) over 5 minutes, and store the resulting mixture at room temperature in an inert atmosphere for 9 minutes. Stirred for 0 minutes. Filter the solid, dilute the filtrate with water (50 mL), and dimethyl(2 × 10⁻¹⁶). Extraction was performed using 0 mL of solution, dried with MgSO4, filtered, and concentrated under reduced pressure. A gray solid was obtained. Next, it was purified using a chromatography column, and 6-(4,5-dichloro-1,1- Dioxide-3-oxoisothiazole-2(3H)-yl)benzyl hexanoate pale yellow It was obtained as a colored oily substance (0.930 g, 44.2%). 1 H NMR (CDCl3), δ 7.36 (m,5H), 5.12 (s, 2H), 3.72 (t, J=7.53 Hz, 2H), 2.73 (t, J=7.50 Hz, 2H), 1.80 (t of t, J= 7,53 Hz, 2H), 1.70 (t of t, J=7.70 Hz), 1.41 (m, 2H); 13 ¹³C NMR (500 MHz, CDCl₃), δ 173.1 (-OC=O-), 154.1 (NC=O), 138.0 (Cl-C-SO2-), 136.0 (c derived from aromatic rings) oque t. ), 130.7 1 (Cl-CC=O), 128.6-128.3 (5 Hc hromatic ), 66.2 (-CH2-O-), 41.1 (-CH2 -N-), 33.9 (-CH2-C=O), 27.9, 26.0, 24.2; R t,1 (in ACN): 2.59; MS ES+ m / z: 405.76 .

[0342] Example 2. 6-(4,5-dichloro-1,1-dioxide-3-oxoisothiazole) -2(3H)-yl)hexanoic acid [ka] Standard procedure for deprotection of benzyl esters: 6-(4,5-dichloro-1,1-dioxide-3-oxoisothiazole-2(3H Benzyl hexanoate (0.930 g, 2.23 mmol, 1 equivalent) is anhydrous DC Diluted in M ​​(11.5 mL). Methanesulfonic acid (1.5 mL, 0.023 mol, (10 equivalents) was added. This solution was stirred at room temperature under an inert atmosphere for 24 hours. Next, this The solution was diluted with DCM and treated with water (50 mL). The organic layer was treated with water (2 × 100 mL), then Next, the solution was extracted with brine (1 x 100 mL). The combined aqueous layer was then mixed with DCM (2 x 100 mL). Extracted using ). The organic layer was dried over MgSO4, concentrated under reduced pressure, and then chromatographically analyzed. Purified using a FI column. 6-(4,5-dichloro-1,1-dioxide-3-o Xisoisothiazole-2(3H)-yl)hexanoic acid was obtained as a clearly colored powder (0.56 3g, 78%). 1 H NMR (500 MHz, CDCl3), δ=3.76 (t, J=7.34 Hz, 2H), 2.38 (t, J= 7.32 Hz, 2H), 1.83 (t of t, J=7.65 Hz, 2H), 1.70 (t of t, J=7.77 Hz, 2H), 1.45 ( t of t, J=7.54 Hz, J=3.31 Hz, 2H); 13 C NMR (500 MHz, CDCl3), δ 178.2 (O=C-OH), 154.2 (NC=O), 138.1 (Cl-C-SO2-), 130.9 (Cl-CC=O), 41.1 (-CH2-N-), 33.4 (-CH2-C =O), 27.9, 25.9, 23.9; R t,1 (in ACN): 2.09; MS ES+ m / z: 315.76.

[0343] I.9. 6-(4-bromo-1,1-dioxide-3-oxoisothiazole-2(3 Benzyl hexanoate (H)-yl) [ka] In CCl4 (40 mL), 6-(1,1-dioxide-3-oxoisothiazole-2 (3H)-yl)benzyl(6-(3-oxoisothiazole-2(3H)- Starting with benzyl hexanoate, the standard procedure for the oxidation of monochlorolide compounds is followed. Therefore, to the solution obtained (3g, 8.89 mmol, 1.00 equivalent), add Br2 (1.2 mL) Add 19.58 mmol (2.2 equivalents) dropwise over 30 minutes while stirring at ambient temperature, and 7 The mixture was stirred overnight at 5°C. This reaction mixture was concentrated under vacuum and diluted with CHCl3 (40 mL). Then, pyridine (0.9g) was added. The resulting solution was stirred at ambient temperature for 30 minutes, and then... Then, the mixture was rapidly cooled by adding 50 ml of saturated NaHCO3 solution. The mixture was washed with saturated sodium carbonate (2 × 50 mL) and 50 mL of brine. The mixture was vacuum concentrated, and the residue was treated with silica gel using ethyl acetate / petroleum ether (1:5). Purified with rum, 0.4g (10.8%) of 6-(4-bromo-1,1-dioxide- 3-Oxoiisothiazole-2(3H)-yl)benzyl hexanoate is used as a pale yellow oily substance. The result obtained was: LC-MS (ES, m / z): 416 [M+H] + , 433[M+NH4]+ ; 1 H-NMR (400 MHz, chloroform) -d) δ 7.42 - 7.28 (m, 2H), 5.12 (s, 1H), 3.69 (t, J = 7.4 Hz, 1H), 2.38 (t, J = 7.4 Hz, 1H), 1.86 - 1.64 (m, 2H), 1.47 - 1.34 (m, 1H).

[0344] Example 3. 6-(4-bromo-1,1-dioxide-3-oxoisothiazole-2( 3H)-yl)hexanoic acid [ka] Dioxane (10 mL) contains 6-(4-bromo-1,1-dioxide-3-oxoiso Thiazol-2(3H)-yl)benzyl hexanoate (1g, 2.40 mmol, 1.0 To a 0 equivalent volume of the solution, 10 mL of 4N HCl was added dropwise while stirring at 0°C. The solution was stirred at room temperature for 2 days. The resulting mixture was vacuum concentrated and dichloromethane (3 × 50) was extracted. Extracted with (mL). The combined organic layers were washed with brine (2 × 100 mL) and anhydrous sodium sulfate. The residue was dried with thorium and concentrated under vacuum. The residue was then treated with DCM / MeOH (10:1). Purified by silica gel column, 100 mg (13%) of 6-(4-bromo-1,1-di Oxido-3-oxoisothiazole-2(3H)-yl)hexanoic acid as a white solid Obtained. LC-MS (ES, m / z): 308[M+NH4] + , 326 / 328[M+H] + ; 1 H-NMR (300 MHz, chloroform m-d) δ 7.58 (s, 1H), 3.75 (t, J = 7.4 Hz, 2H), 2.41 (t, J = 7.4 Hz, 2H), 1.78 (dq, J = 34.6, 7.4 Hz, 4H), 1.47 (t, J = 7.7 Hz, 2H).

[0345] X 1 Example of a synthesis pathway for synthesizing the OR linker: [ka] I.10. 6-(5-(4-cyanophenoxy)-1-oxide-3-oxoisothia Benzyl 2(3H)-yl sol-hexanoate [ka] In a mixture of THF (2 mL) and NaH (0.639 mmol), at 0°C, THF (2 In mL, the solution of 4-hydroxybenzenecarbonitride (76 mg, 0.639 mmol) The liquid was added. After stirring for 30 minutes, 6-(5-chloro-1-oxy) was added to THF (2 mL). Do-3-oxoisothiazole-2(3H)-yl)benzyl hexanoate (250 mg, 0,703 mmol) was added. Next, this reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was diluted with AcOEt, and NH4Cl (10% aqueous solution) was added. Next, the organic phase was added. The product was washed with brine, dried over MgSO4, filtered, and concentrated. Crude product This was purified using a silica gel column with a DCM / MeOH mixture (80 / 20), and 6-( 5-(4-cyanophenoxy)-1-oxide-3-oxoisothiazole-2(3H) -Benzyl hexanoate (210 mg, 75% yield) was obtained as a colorless oil. LC -MS (ES, m / z): 439.0 [M+H]+; 1 ¹H-NMR (300 MHz, chloroform-d) δ 7.79 (m, 2H), 7 .35 (m, 7H), 5.59 (s, 1H), 5.12 (s, 2H), 3.69 (m, 2H), 2.37 (m, 2H), 1.71 (m, 4H) ), 1.39 (m, 2H).

[0346] I.11. 6-(5-(4-cyanophenoxy)-1,1-dioxide-3-oxoi Sothiazol-2(3H)-yl)benzyl hexanoate [ka] 6-(5-(4-cyanophenoxy)-1-oxide-3-oxoisothiazole-2 Starting from benzyl (3H)-yl)hexanoate, a standard for the oxidation of monolith compounds. It was obtained as a colorless oily substance (136 mg, yield 48%) following a standard procedure. LC-MS (ES, m / z ): 455.0 [M+H]+; 1 ¹H-NMR (300 MHz, chloroform-d) δ 7.82 (m, 2H), 7.40 (m, 2H), 7.35 (m, 5H), 5.63 (s, 1H), 5.12 (s, 2H), 3.65 (m, 2H), 2.38 (m, 2H), 1.79 (m, 2H), 1.70 (m, 2H), 1.41 (m, 2H).

[0347] Example 4. 6-(5-(4-cyanophenoxy)-1,1-dioxide-3-oxoi Sothiazol-2(3H)-yl)hexanoic acid [ka] 6-(5-(4-cyanophenoxy)-1,1-dioxide-3-oxoisothiazo Starting from benzyl hydroxy-2(3H)-ylhexanoate, the deprotection of benzyl esters is performed. It was obtained as a white solid (38 mg, yield 35%) following the standard procedure. LC-MS (ES, m / z ): 365.0 [M+H]+; 1H-MR (300 MHz, chloroform-d) 7.82 (m, 2H), 7.41 (m, 2H), 5.6 5 (s, 1H), 3.67 (m, 2H), 2.38 (m, 2H), 1.80 (m, 2H), 1.69 (m, 2H), 1.44 (m, 2H) .

[0348] I.12. 6-(5-methoxy-1-oxide-3-oxoisothiazole-2(3H Benzyl hexanoate (-yl) [ka] In methanol (5 mL) and triethylamine (113 μl, 0.835 mmol) , 6-(5-chloro-1-oxide-3-oxoisothiazole-2(3H)-yl) Mixture of benzyl xanate (270 mg, 0,759 mmol) was stirred at room temperature for 18 hours. Next, volatile substances were removed under vacuum, and the residue was filtered through a silica column using cyclohexane / AcO2. Purified with Et(1 / 1), 6-(5-methoxy-1-oxide-3-oxoisothiazo Benzyl 2(3H)-yl)hexanoate (107 mg, 40%) as a yellow oily substance Obtained. LC-MS (ES, m / z): 352.0 [M+H]+; 1 1H-NMR (300 MHz, chloroform-d) δ 7.36 (m , 5H), 5.57 (s, 1H), 5.11 (s, 2H), 4.04 (s, 3H), 3.60 (m, 2H), 2.37 (m, 2H), 1.7 5 (m, 2H), 1.69 (m, 2H), 1.39 (m, 2H).

[0349] I.13. 6-(5-methoxy-1,1-dioxide-3-oxoisothiazole-2) (3H)-yl)benzyl hexanoate [ka] 6-(5-methoxy-1-oxide-3-oxoisothiazole-2(3H)-yl) Starting with benzyl hexanoate, and following the standard procedure for the oxidation of monochlorolide compounds, It was obtained as a white solid (62 mg, 36% yield). LC-MS (ES, m / z): 368.0 [M+H]+; 1 H-NM R (300 MHz, chloroform-d) δ 7.36 (m, 5H), 5.57 (s, 1H), 5.11 (s, 2H), 4.04 (s , 3H), 3.60 (m, 2H), 2.37 (m, 2H), 1.75 (m, 2H), 1.69 (m, 2H), 1.39 (m, 2H).

[0350] Example 5. 6-(5-methoxy-1,1-dioxide-3-oxoisothiazole-2) (3H)-yl)hexanoic acid [ka] 6-(5-methoxy-1,1-dioxide-3-oxoisothiazole-2(3H)- Starting with benzyl hexanoate, the standard procedure for deprotecting benzyl esters is followed. Therefore, it was obtained as a white solid (37 mg, 67% yield). HRMS (ES, m / z): [M+H] Theoretical value 27 8.0698 versus the measured value of 278.0691; 1 ¹H-NMR (300 MHz, chloroform-d) δ 5.60 (s, 1H), 4. 05 (s, 3H), 3.62 (m, 2H), 2.36 (m, 2H), 1.77 (m, 2H), 1.68 (m, 2H), 1.42 (m, 2H) .

[0351] I.14. 4-(aminomethyl)cyclohexane-1-carboxylate benzyl 4-methyl Lubenzenesulfonate [ka] In toluene (50 mL), 4-(aminomethyl)cyclohexanecarboxylic acid (10 g, 63.61 mmol, 1 equivalent), phenylmethanol (55.03 g, 508.87 ml) (ol, 52.91 mL, 8 equivalents) and TsOH.H2O (12.70 g, 66.79 ml) A mixture of 1.05 moles of toluenesulfonic acid monohydrate is used to recover condensate. To achieve this, a Dean-Stark apparatus was used, and the mixture was stirred at 140°C for 16 hours. It was refluxed for several hours. Later, the reaction mixture became clear. This clear reaction mixture was poured into TBME (500 mL) and then... The resulting white solid was filtered off, washed with TBME (200 mL), and vacuum-dried. 4-(Ammonia (26.) methyl(cyclohexanecarboxylate benzyl); 4-methylbenzenesulfonic acid (26. 6 g, 63.40 mmol, yielded as a white solid (99.68%). 1 1H NMR (400 MH) z, methanol-d4) δ ppm 7.71 (d, J=8.16 Hz, 2 H) 7.28 - 7.40 (m, 5 H) 7.23 (d, J=7.94 Hz, 2 H) 5.11 (s, 2 H) 2.77 (d, J=7.06 Hz, 2 H) 2.29 - 2.39 (m, 4 H) 1.99 - 2.08 (m, 2 H) 1.85 (br d, J=11.25 Hz, 2 H) 1.53 - 1.65 (m, 1 H) 1.43 (qd, J=1 2.97, 3.20 Hz, 2 H) 1.06 (qd, J=12.75, 3.20 Hz, 2 H).

[0352] I.15. 4,4'-(((3,3'-disulfanediylbis(propanoyl))bi (azandiyl) bis(methylene) bis(cyclohexane-1-carboxylic acid dibene) (Gil salt) [ka] In DCM (300 mL), 3-(2-carboxyethyl disulfanyl)propanoic acid ( 6.64g, 31.58mmol, 1 eq), HOBt(9.39g, 69.48mmol (1, 2.2 equivalents) and TEA (12.78 g, 126.33 mmol, 17.58 mL) Add EDCI (13.32 g, 69.48 mmol, 2.2 equivalents) to a solution of 4 equivalents at 0°C. Next, 4-(aminomethyl)cyclohexanecarboxylate benzyl; 4-methyl Ibenzenesulfonic acid (26.5g, 63.17 mmol, 2 equivalents) was added at this temperature. This mixture was stirred at 0-20°C for 4 hours. TLC (Petroleum ether:ethyl acetate = 2: 1. R f A reading of 0.5) indicated that the reaction was complete. This mixture was saturated with NaHCO₃⁻. The mixture was poured into 3 (100 mL) and H2O (100 mL), and the organic layer was separated. The aqueous layer was collected in DCM. Extracted with (200 mL). The combined organic layer was mixed with H2O (100 mL) and brine (100 mL). Washed with (mL), dried with Na2SO4, filtered, and vacuum concentrated. The residue was DCM (50 Dissolve in (mL) and add petroleum ether very slowly until a white precipitate forms. Filter It was then washed with petroleum ether and vacuum dried. 4-[[3-[[3-[(4-benzyl Oxycarbonylcyclohexyl)methylamino]-3-oxopropyl]disulf [Nyl]propanoylamino]methyl]cyclohexanecarboxylate benzyl (19g, 28 0.40 mmol (89.94% yield) was obtained as a white solid; 1 1H NMR (400 MHz, chloro) Holm-d) δ ppm 7.27 - 7.41 (m, 10 H) 6.05 (br s, 2 H) 5.11 (d, J=1.54 Hz, 4 H) 3.10 - 3.17 (m, 4 H) 2.96 - 3.02 (m, 4 H) 2.54 - 2.63 (m, 4 H) 2.30 (td, J=12.2 4, 1.76 Hz, 2 H) 2.04 (br d, J=12.57 Hz, 4 H) 1.85 (br d, J=12.79 Hz, 4 H) 1.38 - 1.54 (m, 6 H) 0.99 (q, J=12.72 Hz, 4 H).

[0353] I.16. 4-((5-chloro-3-oxoisothiazole-2(3H)-yl)methyl (L) Benzyl cyclohexane-1-carboxylate [ka] I.17. 4-((4,5-Dichloro-3-oxoisothiazole-2(3H)-yl )Methyl)cyclohexane-1-carboxylate benzyl [ka] In DCM (100 mL), 4-[[3-[[3-[(4-benzyloxycarbonyl sulfate Chlohexyl)methylamino]-3-oxopropyl]disulfanil]propanoyl [Amino]methyl]cyclohexanecarboxylate benzyl (10g, 14.95 mmol, 1 In a solution of equivalent volume, at 0°C, add sulfuryl chloride (10.09 g, 74.75 mmol, 7.4 7 mL (5 equivalents) was added dropwise. This mixture was stirred at 0-20°C for 12 hours. A clear solution was obtained after the addition of rufuryl. TLC (petroleum ether:ethyl acetate = 2: 1. Rf. (major) = 0.5) indicated that the reaction was complete. This mixture was then converted to H2 Pour into O (30 mL), then into DCM (50 mL) * 2) Extracted. Combined organic layer H2O( Wash with 50 mL of brine, dry with Na2SO4, filter, and vacuum concentrate. It shrunk. Purification by silica gel column revealed 4-[(3-oxoisothiazole-2-I (L)methyl]cyclohexanecarboxylate benzyl (2.1g, 3.42 mmol, yield 1 1.44% (purity 54%) was obtained as a brown solid, and 4-[(5-chloro-3-oxo-iso Thiazol-2-yl)methyl]cyclohexanecarboxylate benzyl (7.1g, 18. 82 mmol, yield 62.96%, purity 97%, was obtained as a grayish-white solid, and 4-[(4,5 -Dichloro-3-oxoisothiazol-2-yl)methyl]cyclohexanecarbon Benzyl acid (0.4g, 869.31μmol, yield 2.91%, purity 87%) was used to make brown oil. It was obtained as a substance.

[0354] I.18. 4-((5-chloro-1,1-dioxide-3-oxoisothiazole-2 (3H)-yl)methyl)cyclohexane-1-carboxylate benzyl [ka] In H2O (20 mL), 4-[(5-chloro-3-oxoisothiazol-2-yl) ) Methyl]cyclohexanecarboxylate benzyl (2g, 5.01 mmol, 1 equivalent), A A mixture of CN (10 mL) and DCM (10 mL) contains RuCl3.H2O(22.5 (8 mg, 100.14 μmol, 0.02 equivalents) and NaIO4 (6.43 g, 30. (0.4 mmol, 1.66 mL, 6 equivalents) was added all at once at 0°C under N2 conditions. Next, this mixture The mixture was heated to 20°C and stirred for 16 hours. TLC indicated that the reaction was complete. Petroleum ether:ethyl acetate = 2:1, Rf-p1 = 0.6). Filter this mixture and obtain the filtrate. The solution was concentrated with a nitrogen stream, the solid that reappeared was filtered, and the filtrate was concentrated with nitrogen. TLC (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel) Purified by (petroleum ether:ethyl acetate = 2:1), 4-[(5-chloro-1,1, 3-Trioxo-isothiazole-2-yl)methyl]cyclohexanecarboxylate benzyl A white solid (1.4g, 3.17 mmol, yield 63.25%, purity 90%) was obtained. Ta.

[0355] Example 6. 4-((5-chloro-1,1-dioxide-3-oxoisothiazole-2 (3H)-yl)methyl)cyclohexane-1-carboxylic acid [ka] In DCM (5 mL), 4-[(5-chloro-1,1,3-trioxo-isothiazole [-2-yl]methyl]cyclohexanecarboxylate benzyl (0.1g, 226.20μm A mixture of (ol, 1 equivalent) contains MsOH (217.39 mg, 2.26 mmol, 161. (0.3 μL, 10 equivalents) was added all at once at 30°C under N2 conditions. The mixture was stirred at 30°C for 16 hours. Mixed. TLC showed that the reaction was complete. LCMS(ET17992-54- P1A) indicated that the target MS was detected. Pour this mixture into ice water (5 mL). The mixture was stirred for 5 minutes. The aqueous phase was converted to DCM (3 mL). * Extracted in step 2). The combined organic phase was brined. Washed with (3 mL), dried with anhydrous Na2SO4, filtered, and vacuum concentrated. The residue was separated. TLC (Column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel) , purified with petroleum ether: / ethyl acetate (2:1), 4-[(5-chloro-1,1, 3-Trioxo-isothiazole-2-yl)methyl]cyclohexanecarboxylic acid (0. 0.2 g, 64.99 μmol, yield 28.73%) was obtained as a colorless oily substance; LC-MS (E S, m / z): 306.0 [MH] - ;1 H-NMR (300 MHz, DMSO-D6) δ 12.01 (bs, 1H), 7.62 (s, 1H) , 3.45 (m, 2H), 2.11 (m, 1H), 1.90 (m, 2H), 1.76 (m, 2H), 1.73 (m, 1H), 1.23 (m, 2H), 0.96 (m, 2H).

[0356] I.19 4-((4,5-Dichloro-1,1-dioxide-3-oxoisothiazole -2(3H)-yl)methyl)cyclohexane-1-carboxylate benzyl [ka] In H2O (20 mL), DCM (10 mL), and ACN (10 mL), 4-[(4, 5-Dichloro-3-oxoisothiazole-2-yl)methyl]cyclohexanecarb A mixture of benzyl phosphate (1.2g, 2.70 mmol, 1 equivalent) and RuCl3.H2O (12.16 mg, 53.96 μmol, 0.02 equivalents) and NaIO4 (3.46 g (16.19 mmol, 896.96 μL, 6 equivalents) was added all at once at 0°C under N2 conditions. The mixture was stirred at 20°C for 2 hours. TLC showed that the reaction was complete (petroleum ester). (ethyl acetate = 2:1, Rf-p1 = 0.7). Add this mixture to ice water (30 mL). Pour in and stir for 5 minutes. Add ethyl acetate (20 mL) to the aqueous phase. * Extracted in step 2). Combined organic phase. Washed with brine (20 mL), dried with anhydrous Na2SO4, filtered, and vacuum concentrated. The residue was subjected to silica gel chromatography (column height: 250 mm, diameter: 100 mm, 1 00-200 mesh silica gel, petroleum ether:ethyl acetate (5:1-3:1) Purified, 4-[(4,5-dichloro-1,1,3-trioxoisothiazole-2- (L) Methyl cyclohexanecarboxylate benzyl (0.8g, 1.67 mmol, yield 6 A 1.73% solution with a purity of 90% was obtained as a white solid.

[0357] Example 7. 4-((4,5-dichloro-1,1-dioxide-3-oxoisothiazo Lu-2(3H)-yl)methyl)cyclohexane-1-carboxylic acid [ka] In DCM (20 mL), 4-[(4,5-dichloro-1,1,3-trioxo-isoth Azole-2-yl methyl cyclohexanecarboxylate benzyl (0.8g, 1.67 A mixture of mmol (1 equivalent) and MsOH (1.60 g, 16.65 mmol, 1.19 The solution (10 equivalents, mL) was added all at once at 30°C under N2 conditions. The mixture was stirred at 30°C for 16 hours. The LC-MS showed that the reaction was complete. Pour this mixture into ice water (5 mL). The solution was concentrated under reduced pressure, after which a solid appeared. This solution was filtered and HCl (2 mL) was removed. * 3 ) is ground, the filtered cake is vacuum dried, and 4-[(4,5-dichloro-1,1,3- Trioxo-isothiazole-2-yl)methyl]cyclohexanecarboxylic acid (0.17 0 g, 486.86 μmol, yield 29.23%, purity 98% was obtained as a white solid; LC-MS (ES, m / z): 364.0 [M+Na] + ; 1 H-NMR (300 MHz, DMSO-D6) δ 11.99 (bs, 1H), 3. 49 (m, 2H), 2.11 (m, 1H), 1.88 (m, 2H), 1.79 (m, 2H), 1.66 (m, 1H), 1.23 (m, 2H) , 0.96 (m, 2H).

[0358] I.20 3,3'-(((3,3'-disulfanediylbis(propanoyl))bis (Azandiyl))bis(4,1-phenylene))dibenzyl dipropionate [ka] In DCM (50 mL), 3-(2-carboxyethyl disulfanyl)propanoic acid (6 51.56 mg, 3.10 mmol, 1 eq), HOBt (921.15 mg, 6.82 mmol, 2.2 equivalents) and TEA (1.25 g, 12.39 mmol, 1.73 mL) Add EDCI (1.31 g, 6.82 mmol, 2.2 equivalents) to a solution of 4 equivalents at 0°C. Added. Next, 4-(aminomethyl)cyclohexanecarboxylate benzyl; 4-methylbe Sulfonic acid (2.6g, 6.20 mmol, 2 equivalents) was added at this temperature. The mixture was stirred at 0-20°C for 12 hours. TLC (petroleum ether:ethyl acetate = 2:1, R f=0.5) indicated that the reaction was complete. This mixture was saturated with NaHCO3(2 The layers were poured into DCM (50 mL) and H2O (20 mL) and the organic layer was separated. Extracted using ). The combined organic layers were washed with H2O (50 mL) and brine (50 mL). The solution was dried with Na2SO4, filtered, and vacuum concentrated. 4-[[3-[[3-[(4-benzyl [Oxycarbonylcyclohexyl)methylamino]-3-oxopropyl]disulf [Anylon]propanoylamino]methyl]cyclohexanecarboxylate benzyl (1.1g, 1.64 mmol (53.07% yield) was obtained as a white solid; 1 1H NMR (400 MHz, Chromium Loform-d) δ ppm 7.28 - 7.41 (m,°10 H) 5.96 (br s, 2 H) 5.10 (s, 4 H) 3.13 (t , J=6.39 Hz, 4 H) 2.98 (t, J=6.84 Hz, 4 H) 2.57 (t, J=6.95 Hz, 4 H) 2.23 - 2.34 (m, 2 H) 2.03 (br d, J=12.79 Hz, 4 H) 1.84 (br d, J=12.13 Hz, 4 H) 1.37 - 1.63 ( m,°6 H) 0.91 - 1.05 (m, 4 H).

[0359] I.21. 3-(4-(5-chloro-3-oxoisothiazole-2(3H)-yl) Benzyl phenylpropanoate [ka] In DCM (200 mL), 3-[4-[3-[[3-[4-(3-benzyloxy-3 -Oxopropyl)anilino]-3-oxopropyl]disulfanil]propano Solubility of benzyl aminophenyl propanoate (10g, 14.60 mmol, 1 equivalent) Add sulfuryl chloride (5.91 g, 43.80 mmol, 4.38 mL, 3 equivalents) to the solution. The solution was added dropwise at 5°C under N2 conditions. The solution was stirred at 25°C for 8 hours. Sulfuryl chloride was then added. The solution's color changed from colorless to black, and then to yellow after 1 hour. (TLC) Petroleum ether:ethyl acetate = 2:1, Rf = 0.60) The starting material is consumed, and two This indicated that new spots had formed. Pour the residue into ice water (200 ml), then, The DCM was removed by vacuum concentration. After concentration, the aqueous phase was treated with ethyl acetate (200 mL). * 3) Extraction, Next, add the combined organic phase to water (200 mL) * 1) Wash and dry with anhydrous sodium 2SO4. The residue was filtered and vacuum concentrated. The residue was subjected to column chromatography (SiO2, petroleum ether: Purified with ethyl acetate (5:1~1:1), 3-[4-(3-oxoisothiazole- Benzyl 2-yl)phenyl]propanoate (2.5g, 7.37 mmol, yield 50.4%) 7% obtained; 1 ¹H NMR (400MHz, methanol-d4) δ = 8.56 (d, J=6.2 Hz, 1H), 7.44 - 7 .39 (m, 2H), 7.37 - 7.26 (m, 7H), 6.30 (d, J=6.4 Hz, 1H), 5.09 (s, 2H), 2.98 (t, J=7.4 Hz, 2H), 2.78 - 2.66 (m, 2H); and 3-[4-(5-chloro-3-oxo- Isothiazol-2-yl)phenyl]propanoate benzyl (5g, 13.37mmol) A yellow solid was obtained with a yield of 91.60%. 1 1H NMR (ET17992-22-P It was identified as ET17992-22-P2 by 2A). 1 1H NMR (400MHz, methanol-d 4) δ = 7.42 - 7.36 (m, 2H), 7.35 - 7.25 (m, 7H), 6.49 - 6.45 (m, 1H), 5.08 (s, 2H), 2.99 - 2.91 (m, 2H), 2.69 (t, J=7.5 Hz, 2H).

[0360] I.22. 3-(4-(5-chloro-1,1-dioxide-3-oxoisothiazole Benzyl 2(3H)-yl(phenyl)propanoate [ka] In H2O (12 mL), DCM (6 mL), and ACN (6 mL), 3-[4-(5- Benzyl chloro-3-oxoisothiazol-2-yl)phenyl]propanoate (1. A mixture of 4g, 3.74 mmol, 1 equivalent contains NaIO4 (4.81g, 22.47m Add (1.25 mL, 6 equivalents) of the mixture all at once at 25°C, then add the mixture to N2 for 3 minutes. The system was purged multiple times. Next, RuCl3.H2O(42.21 mg, 187.24 μmol, 0 (0.05 equivalents) was added under N2 conditions. This mixture was stirred at 25°C for 12 hours. The solution became cloudy and gray. The residue was poured into ethyl acetate (100 ml) and then filtered. The liquid was vacuum concentrated. The residue was subjected to column chromatography (SiO2, petroleum ether: acetate). Purified by (10:1~4:1), 3-[4-(5-chloro-1,1,3-trio Xo-isothiazol-2-yl)phenyl]benzyl propanoate (460 mg, 1.0 8 mmol was obtained as a yellow solid with a yield of 28.75% and a purity of 95%. 1 H NMR (E It was confirmed to be ET17992-63-P1 by T17992-63-P1A. 1 1H NMR (400MHz, chloroform-d) δ = 7.41 - 7.30 (m, 7H), 6.84 (s, 1H), 5.13 (s, 2H), 3.04 (t, J=7.6 Hz, 2H), 2.72 (t, J=7.7 Hz, 2H).

[0361] Example 8. 3-(4-(5-chloro-1,1-dioxide-3-oxoisothiazole -2(3H)-yl)phenyl)propanoic acid [ka] In DCM (15 mL), 3-[4-(5-chloro-1,1,3-trioxoisothia Benzyl zole-2-yl)phenyl]propanoate (460 mg, 1.13 mmol, 1 In a solution of equivalent volume, add methanesulfonic acid (1.09 g, 11.33 mmol, 806.87 μg) (L, 10 equivalents) was added dropwise at 10°C. Next, this solution was heated to 35°C and stirred for 12 hours. The residue was added to water (15ml). * 3) Wash, dry with anhydrous Na2SO4, filter, vacuum concentrate It shrunk. The residue was poured into water (20 ml) and then filtered. The filtered cake was refrigerated in DCM (5 ml). Dissolve in ), then pour petroleum ether (30 ml) into this residue, and leave the solution at 10°C for 2 Stir for minutes, then filter, and vacuum dry the filtered cake, 3-[4-(5-chloro-1, 1,3-Trioxoisothiazol-2-yl)phenyl]propanoic acid (162 mg, 501.81 μmol, yield 44.27%, purity 97.8% was obtained as a white solid; L C-MS (ES, m / z): 313.9 [MH] - ; 1 H-NMR (300 MHz, DMSO-D6) δ 12.18 (bs, 1H), 7.81 (s, 1H), 7.45 (m, 2H), 7.37 (m, 2H), 2.89 (m, 2H), 2.58 (m, 2H).

[0362] I.23. 3-(4-(4,5-Dichloro-3-oxoisothiazole-2(3H)- Benzyl phenylpropanoate [ka] In H2O (20 mL), ACN (10 mL), and DCM (10 mL), 4-[(3- Oxoisothiazol-2-yl)methyl]cyclohexanecarboxylate benzyl (1.2 A mixture of (g, 3.32 mmol, 1 equivalent) contains RCl3.H2O (14.95 mg, 66 (0.33 μmol, 0.02 equivalents) and NaIO4 (4.26 g, 19.90 mmol, 1.10 mL (6 equivalents) was added all at once at 0°C under N2 conditions. Next, this mixture was added at 20°C. It was heated and stirred for 16 hours. This mixture was filtered, and the filtrate was concentrated with a nitrogen stream, and the solid was again The residue was extracted, filtered, and the filtrate was concentrated with nitrogen. The residue was then separated by preparative TLC (column height: 250 mm). Diameter: 100mm, 100-200 mesh silica gel, petroleum ether:ethyl acetate = 2 :1) Purified by 4-[(1,1,3-trioxoisothiazole-2-yl)methyl [L] Benzyl cyclohexanecarboxylate (0.45 g, 1.11 mmol, yield 33.6%) A 0% concentration, 90% purity product was obtained as a colorless oily substance; 1 ¹H NMR (400MHz, chloroform-d) δ = 7.53 (dd, J=3.2, 8.7 Hz, 2H), 7.17 (ddd, J=3.2, 7.6, 9.0 Hz, 2H), 6.97 - 6.89 ( m, 2H), 5.56 (br s, 1H), 4.12 - 4.06 (m, 4H), 3.92 (s, 5H), 3.55 (q, J=5.1 Hz, 5 H).

[0363] I.24. 3-(4-(4,5-Dichloro-1,1-dioxide-3-oxoisothia Benzyl 2(3H)-yl(phenyl)propanoate [ka] 3-[4-(4,5- Dichloro-3-oxo-isothiazol-2-yl)phenyl]propanoate benzyl (6 (50 mg, 1.59 mmol, 1 equivalent) and NaIO4 (1.36 g, 6.37 mmol) A mixture of 1, 352.86 μL, 4 equivalents of RuCl3, H2O (7.18 mg, 31 0.84 μmol (0.02 equivalents) was added all at once at 0°C under N2 conditions. This mixture was then heated at 20°C. The mixture was stirred for 2 hours. The residue was then mixed with ethyl acetate (30 mL). * Extracted in step 2). The combined organic phase was anhydrous. The residue was dried with Na2SO4, filtered, and vacuum concentrated. The residue was then subjected to column chromatography (S 3-[4-(4 ,5-Dichloro-1,1,3-Trioxoisothiazole-2-yl)phenyl]pro Benzyl panate (180 mg, yield 25.68%) was obtained as a yellow solid; 1 1H NMR (400M Hz, chloroform (d) δ = 7.43 - 7.29 (m, 9H), 5.13 (s, 2H), 3.05 (t, J=7.6 Hz, 2 H), 2.73 (t, J=7.6 Hz, 2H).

[0364] Example 9. 3-(4-(4,5-dichloro-1,1-dioxide-3-oxoisothia Zole-2(3H)-yl)phenyl)propanoic acid [ka] In DCM (5 mL), 3-[4-(4,5-dichloro-1,1,3-trioxo-iso Benzyl thiazole-2-yl)phenyl]propanoate (180 mg, 408.81 μm) In a solution of ol (1 equivalent), methanesulfonic acid (392.89 mg, 4.09 mmol, 2) 91.03 μL (10 equivalents) was added dropwise at 10°C. This solution was stirred at 35°C for 12 hours. The residue was poured into water (5 ml) and then filtered. The filtered cake was then poured into water (10 ml) * 3) Wash The residue was purified and then vacuum-dried. The residue was treated with dichloromethane:methanol (5.5 ml, v / Dissolve in v=10:1, then perform preparative TLC (ethyl acetate, Rf=0.26) Purified, 3-[4-(4,5-dichloro-1,1,3-trioxo-isothiazole-2 -yl)phenyl]propanoic acid (57.64 mg, 156.83 μmol, yield 38.3 A 6% concentration with a purity of 95.278% was obtained as a white solid; LC-MS (ES, m / z): 347.9 [MH]-; 1 H NMR (400MHz, DMSO-d6) δ = 12.22 (br s, 1H), 7.51 - 7.40 (m, 4H), 2.90 (br t, J=7.6 Hz, 2H), 2.60 (br t, J=7.6 Hz, 2H).

[0365] I.25. 4,4'-((3,3'-disulfanediylbis(propanoyl))bis (Azandiyl) Dibenzyl Dibenzoate [ka] In DMF (120 mL), 3-(2-carboxyethyl disulfanyl)propanoic acid ( 6.01g, 28.60 mmol, 1 equivalent) and pyridine (14.93g, 188.7 Add EDCI (12.06 g, 62.5g) to a solution of 7 mmol, 15.24 mL, 6.6 equivalents. (92 mmol, 2.2 equivalents) and benzyl 4-aminobenzoate (13 g, 57.20 mg) (2 equivalents of mol) was added at 10°C. Next, this mixture was stirred at 50°C for 12 hours. The residue was poured into ice water (200 mL) and stirred for 20 minutes. The aqueous phase was converted to ethyl acetate (200 mL) * 3 The organic phase was extracted using ) and the combined organic phase was saturated with NaCl (200 mL). * 3) Wash, anhydrous Na2 The residue was dried with SO4, filtered, and vacuum concentrated. Next, the residue was extracted into petroleum ether:DCM=50: A solid was obtained by recrystallizing from step 1. This solid was washed three times with petroleum (150 ml). * 3) Next Then, vacuum-dried, 4-[3-[[3-(4-benzyloxycarbonylanilino)-3 -Oxo-propyl]disulfanyl]propanoylamino]benzyl benzoate (14g The crude product was obtained as a white solid; 1 H NMR (400MHz, DMSO-d6) δ = 10.37 (s, 2H), 8 .02 - 7.83 (m, 4H), 7.72 (d, J=8.8 Hz, 4H), 7.48 - 7.32 (m, 10H), 5.31 (s, 4H), 3.05 - 2.98 (m, 4H), 2.81 - 2.75 (m, 4H).

[0366] I.26. 4-(5-chloro-3-oxoisothiazole-2(3H)-yl)benzoic acid benzyl acid [ka] I.27. 4-(4,5-dichloro-3-oxoisothiazole-2(3H)-yl) Benzyl benzoate [ka] In DCM (120 mL), 4-[3-[[3-(4-benzyloxycarbonylanili [(n)-3-oxo-propyl]disulfanyl]propanoylamino]benzyl benzoate (9.4g, 14.95 mmol, 1 equivalent) solution, sulfuryl chloride (10.09g, 74.75 mmol, 7.47 mL, 5 equivalents were added dropwise at 0°C. This mixture was then 0-20 The mixture was stirred at °C for 12 hours. After stirring for a few minutes, the mixture became clear. TLC (Petroleum E The reaction was complete when the ethyl acetate (ethyl acetate:ethyl acetate = 2:1) was mixed with H2O. Pour into (200mL), DCM (200mL * Extracted in step 2). Combined organic layer H2O (200mL * 2) The residue was washed, dried with Na2SO4, filtered, and vacuum concentrated. Column chromatography using silica gel (petroleum ether:ethyl acetate = 5:1 to 1:1) - Purified by -, 4-(3-oxoisothiazol-2-yl)benzyl benzoate (1. 2 g, 3.43 mmol, yield 11.47%, purity 89% was obtained as a grayish-white solid; 1 HN MR (400 MHz, chloroform-d) δ ppm 8.09 - 8.27 (m, 3 H) 7.63 - 7.82 (m, 2 H) 7. 32 - 7.52 (m, 5 H) 6.34 (br d, J=6.36 Hz, 1 H) 5.39 (s, 2 H); 4-(5-chloro- 3-Oxo-isothiazol-2-yl)benzyl benzoate (3.5g, 10.01mm) ol was obtained as a grayish-white solid with a yield of 33.49% and a purity of 98.92%; 1 1H NMR (400 MHz, Chloroform - d) δ ppm 8.15 (d, J=8.60 Hz, 2 H) 7.69 (d, J=8.82 Hz, 2 H) 7.33 - 7.49 (m, 4 H) 6.38 (s, 1 H) 5.38 (s, 2 H) and 4-(4,5-dichloro-3-oxy (So-isothiazol-2-yl)benzyl benzoate (2.8g, 7.19 mmol, yield) A 24.06% solution with a purity of 97.68% was obtained as a grayish-white solid; 1 1H NMR (400 MHz, chloro) Holm-d) δ ppm 8.18 (d, J=8.60 Hz, 2 H) 7.71 (d, J=8.60 Hz, 2 H) 7.33 - 7.50 ( m, 4 H) 5.39 (s, 2 H).

[0367] I.28. 4-(5-chloro-1,1-dioxide-3-oxoisothiazole-2( Benzyl benzoate (3H)-yl [ka] In H2O (10 mL), ACN (5 mL), and DCM (5 mL), 4-(5-chloro Benzyl benzoate (-3-oxo-isothiazol-2-yl) (1g, 2.89 mmol) A mixture of 1 equivalent of RuCl3H2O(13.04 mg, 57.84 μmol, 0 (0.02 equivalents) and NaIO4 (2.47 g, 11.57 mmol, 640.97 μL, (4 equivalents) was added all at once at 0°C under N2 conditions. Next, this mixture was heated to 20°C and stirred for 2 hours. Mixed. Filter the residue and pour the filtrate into water (40 ml). Add ethyl acetate (50 ml) to the aqueous phase. * 1) Extracted. The organic phase was saturated with NaCl (30 mL). * 3) Wash with anhydrous Na2SO4 The residue was dried, filtered, and vacuum concentrated. The residue was then subjected to column chromatography (SiO2, petroleum ester). Purified by ethyl acetate (15:1~5:1), 4-(5-chloro-1,1,3 -Trioxo-isothiazol-2-yl)benzyl benzoate (500mg, 1.32mg) A quantity of mol, yielding 45.77%, was obtained as a yellow oily substance; 1 H NMR (400 MHz, chloroform-d) ) δ = 8.27 - 8.21 (m, 2H), 7.61 - 7.55 (m, 2H), 7.49 - 7.34 (m, 5H), 6.87 (s, 1 H), 5.40 (s, 2H).

[0368] Example 10. 4-(5-chloro-1,1-dioxide-3-oxoisothiazole-2) (3H)-yl)benzoic acid [ka] DCM (20 mL) contains 4-(5-chloro-1,1,3-trioxo-isothiazole) A solution of benzyl benzoate (450 mg, 1.19 mmol, 1 equivalent) contains... Tansulfonic acid (1.14 g, 11.91 mmol, 847.94 μL, 10 equivalents) The solution was added dropwise at 0°C. The solution was stirred at 35°C for 10 hours. The residue was vacuum concentrated to remove DCM. Next, the residue was dissolved in ethyl acetate (10 ml), and then the organic phase was dissolved in water (20 ml). * 5) Washed, dried with anhydrous Na2SO4, filtered, and vacuum concentrated. The residue was methanol. Dissolve in 3 ml of ru, then pour 30 ml of petroleum ether into the residue, and this solution Stir at 10°C for 2 minutes, then filter, and vacuum dry the filtered cake, then 4-(5-chloro -1,1,3-trioxoisothiazol-2-yl)benzoic acid (112.56 mg, 379.48 μmol was obtained as a white solid with a yield of 31.86% and a purity of 96.986%. ; LC-MS (ES, m / z): 285.9 [MH]-; 1 H NMR (400MHz, DMSO-d6) δ = 13.33 (br s, 1H) , 8.17 - 8.11 (m, 2H), 7.87 (d, J=1.5 Hz, 1H), 7.67 - 7.61 (m, 2H).

[0369] I.29. 4-(4,5-dichloro-1,1-dioxide-3-oxoisothiazole) -2(3H)-yl)benzyl benzoate [ka] In H2O (10 mL), ACN (5 mL), and DCM (5 mL), 4-(4,5-di Benzyl chlorobenzoate (1g, 2.63mg) A mixture of mol, 1 equivalent, contains RuCl3.H2O (11.86 mg, 52.60 μmol) (1, 0.02 equivalents) and NaIO4 (2.25 g, 10.52 mmol, 582.91 (μL, 4 equivalents) was added all at once at 0°C under N2 conditions. Next, this mixture was heated to 20°C. The mixture was stirred for a certain amount of time. The residue was filtered, and the filtrate was concentrated under vacuum. The residue was subjected to column chromatography. Purified with SiO2, petroleum ether:ethyl acetate (20:1~5:1), and 4-(4,5 -Dichloro-1,1,3-trioxoisothiazol-2-yl)benzyl benzoate ( 130 mg, 315.35 μmol, yield 11.99%) was obtained as a white solid; 1 1H NMR (400MHz, chloroform-d) δ = 8.25 (d, J=8.6 Hz, 2H), 7.58 (d, J=8.8 Hz, 2H), 7 .49 - 7.34 (m, 5H), 5.41 (s, 2H).

[0370] Example 11. 4-(4,5-dichloro-1,1-dioxide-3-oxoisothiazo Ru-2(3H)-yl)benzoic acid [ka] In DCM (5 mL), 4-(4,5-dichloro-1,1,3-trioxo-isothiazo Dissolved in benzyl benzoate (130 mg, 315.35 μmol, 1 equivalent) In the solution, add methanesulfonic acid (303.07 mg, 3.15 mmol, 224.49 μL, 1 (0 equivalents) was added dropwise at 10°C. This solution was stirred at 10°C for 5 minutes, and then this solution was 3 The solution was heated to 5°C and stirred for 10 hours. The color of the solution changed from colorless to yellow. Poured into water (20 ml) and then filtered. The filtered cake was then poured into water (10 ml) * 3) and DC M (10ml) * 3) Each was washed three times. Next, the filtered cake was vacuum dried. Residue Disperse with DCM (10 ml), then pour petroleum ether (30 ml) onto the residue, Stir the mixture at 10°C for 2 minutes, then filter it, vacuum dry the filtered cake, and 4-( 4,5-Dichloro-1,1,3-trioxoisothiazol-2-yl)benzoic acid (9 3.4 mg, 282.97 μmol, yield 89.73%, purity 97.590% formed a white solid. Obtained as a whole; LC-MS (ES, m / z): 319.9 [MH]-; 1H NMR (400MHz, DMSO-d6) δ = 8.16 (d, J=8.4 Hz, 2H), 7.68 (d, J=8.4 Hz, 2H).

[0371] I.30. 3-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)propanoic acid Benzyl 4-methylbenzenesulfonate [ka] In toluene (30 mL), 3-[2-[2-(2-aminoethoxy)ethoxy]ethoxy [C]propanoic acid (4g, 18.08 mmol, 1 equivalent), phenylmethanol (15.6 4g, 144.63 mmol, 15.04 mL, 8 equivalents) and TsOH.H2O(3. A mixture of 61g, 18.98 mmol, and 1.05 equivalents is used to collect condensate. The mixture was stirred at 140°C for 8 hours using a Neon-Stark apparatus. After refluxing the mixture for several hours, It became transparent. TLC (dichloromethane:methanol = 10:1, Rf = 0.3) This indicated that the reaction was complete. This clear reaction mixture contains TBME: petroleum ether (1 Poured into 1, 50 mL of water and removed the clear solution. The residue was TBME: petroleum ether (1: Washed twice with 1.50 mL and vacuum-dried. Crude product 3-[2-[2-(2-amino Ethoxy)Ethoxy]Ethoxy]Benzyl propanoate; 4-methylbenzenesulfonic acid ( 8.9 g (crude product) was obtained as a yellow oily substance; 1 1H NMR (400 MHz, chloroform-d) δ ppm 7.76 (br d, J=8.07 Hz, 2 H) 7.33 - 7.38 (m, 5 H) 7.15 (d, J=7.95 Hz, 2 H) 5. 11 (s, 2 H) 3.72 (q, J=6.11 Hz, 4 H) 3.53 - 3.64 (m, 8 H) 3.11 - 3.24 (m, 2 H) 2 .53 - 2.69 (m, 2 H) 2.30 - 2.41 (m, 1 H) 2.34 (s, 3 H).

[0372] I.31. 3-[2-[2-[2-[3-[[3-[2-[2-[2-(3-benzyl Oxy-3-oxo-propoxy)ethoxy]ethoxy]ethylamino]-3-oxop Ropil Disulfanyl Propanoylamino Ethoxy Ethoxy Ethoxy Propa benzyl phosphate [ka] In DCM (100 mL), 3-(2-carboxyethyl disulfanyl)propanoic acid ( 1.90g, 9.04mmol, 1 eq), HOBt(2.69g, 19.88mmol (2.2 equivalents) and TEA (4.57 g, 45.18 mmol, 6.29 mL, 5 equivalents) Add EDCI (3.81g, 19.88 mmol, 2.2 equivalents) to the mixture at 20°C. Next, add 3-[2-[2-(2-aminoethoxy)ethoxy]ethoxy] to the above solution. Benzyl propanoate; 4-methylbenzenesulfonic acid (8.74g, 18.07mmol) (2 equivalents) was added. This mixture was stirred at 20°C for 12 hours. TLC (petroleum ether: The presence of ethyl acetate (2:1, Rf=0.25) indicated that the reaction was complete. The solution was then concentrated under reduced pressure and vacuum-dried. The residue was poured into H2O (20 mL). Extracted with ethyl acetate (50 mL). The organic layer was dried with Na2SO4, filtered, and vacuum-sealed. The solution was concentrated. The crude product was fractionated by TLC (petroleum ether:ethyl acetate = 1:1, Rf = 0.5). Purified by 3-[2-[2-[2-[3-[[3-[2-[2-[2-(3-benzyl Luoxy-3-oxo-propoxy)ethoxy]ethoxy]ethylamino]-3-oxo -Propyl]disulfanyl]propanoylamino]ethoxy]ethoxy]ethoxy]p Benzyl ropanoate (6.52g, 7.94 mmol, yield 87.81%, purity 97%) It was added as a yellow oily substance; 1 ¹H NMR (400 MHz, chloroform-d) δ ppm 7.29 - 7.42 (m, 10 H) 6.42 (br s, 2 H) 5.15 (s, 4 H) 3.79 (t, J=6.42 Hz, 4 H) 3.53 - 3.69 (m, 18 H) 2.92 - 3.00 (m, 4 H) 2.62 - 2.71 (m, 4 H) 2.53 - 2.62 (m, 4 H).

[0373] I.32. 3-(2-(2-(2-(5-chloro-3-oxoisothiazole-2(3 H)-Iyl)ethoxy)ethoxy)ethoxy)benzyl propanoate [ka] 3-[2-[2-[2-[3-[[3-[2-[2-[2-( 3-benzyloxy-3-oxopropoxy)ethoxy]ethoxy]ethylamino]- 3-Oxopropyl]disulfanyl]propanoylamino]ethoxy]ethoxy]eth In a solution of benzyl xypropanoate (6.4 g, 8.03 mmol, 1 equivalent), at 0°C, In DCM (10 mL), sulfuryl chloride (4.34 g, 32.12 mmol, 3.21 mL) A solution (4 equivalents, L) was added dropwise. This mixture was stirred at 0-10°C for 12 hours. TLC ( Petroleum ether:ethyl acetate = 0:1, R f (=0.15, 0.35) indicates that the reaction is complete. This demonstrated that... Pour this mixture into ice / water (100 mL) and add DCM (200 mL) * 2) Extracted using H2O (100mL). The combined organic layer was then converted to H2O (100mL). * 2) Wash with brine (100 mL) The solution was purified and dried with Na2SO4. It was filtered and vacuum concentrated. The residue was column-concentrated using silica gel. Purified by chromatography (petroleum ether:ethyl acetate = 1:1 to 0:1), 3- [2-[2-[2-(3-oxoisothiazol-2-yl)ethoxy]ethoxy]eth Benzyl xylpropanoate (820 mg, 1.66 mmol, yield 10.33%, purity 8) 0% is obtained as a brown oily substance; 1 ¹H NMR (400 MHz, chloroform-d) δ ppm 8.06 (d, J=6 .17 Hz, 1 H) 7.30 - 7.41 (m, 5 H) 6.24 (d, J=6.39 Hz, 1 H) 5.15 (s, 2 H) 3.95 - 4.03 (m, 2 H) 3.79 (t, J=6.39 Hz, 2 H) 3.68 - 3.75 (m, 2 H) 3.59 - 3.68 (m, 8 H) 2.63 - 2.70 (m, 2 H) and 3-[2-[2-[2-(5-chloro-3-oxo-iso Thiazol-2-yl)ethoxy]ethoxy]ethoxy]benzyl propanoate (2.5g) 4.30 mmol, yield 26.79%, purity 74%, was obtained as a colorless oily substance; 1 HN MR (400 MHz, chloroform-d) δ ppm 7.28 - 7.43 (m, 5 H) 6.25 (s, 1 H) 5.15 (s, 2 H) 3.92 - 3.98 (m, 2 H) 3.77 - 3.81 (m, 2 H) 3.59 - 3. 71 (m, 10 H) 2.66 (t, J=6.39 Hz, 2 H).

[0374] I.33. 3-(2-(2-(2-(5-chloro-1,1-dioxide-3-oxoi Sothiazole-2(3H)-yl)ethoxy)ethoxy)ethoxy)benzyl propanoate [ka] 3-[2-[2- [2-(5-chloro-3-oxo-isothiazole-2-yl)ethoxy]ethoxy] Benzyl toxypropanoate (1g, 2.33 mmol, 1 equivalent) and NaIO4 (1 A mixture of 0.99 g, 9.30 mmol, 515.57 μL, and 4 equivalents of RuCl3H is added. 2O (26.22 mg, 116.30 μmol, 0.05 equivalents) was incubated at 20°C under N2 conditions once It was added to [the mixture]. Next, this mixture was stirred at 20°C for 1 hour. The residue was then added to ethyl acetate (20 mL). * 2) Extraction was performed. The combined organic phase was dried with anhydrous Na2SO4, filtered, and vacuum concentrated. The residue was subjected to column chromatography (SiO2, petroleum ether:ethyl acetate = 5:1~1 :1) Purified by 3-[2-[2-[2-(5-chloro-1,1,3-trioxo- Isothiazol-2-yl)ethoxy]ethoxy]ethoxy]benzyl propanoate (56 0 mg, 1.21 mmol, yield 52.12%, purity 100%) was obtained as a purple oily substance. Ta; 1 ¹H NMR (400MHz, chloroform-d) δ = 7.42 - 7.29 (m, 5H), 6.70 (s, 1H), 5.15 (s, 2H), 3.92 - 3.86 (m, 2H), 3.77 (td, J=6.0, 11.9 Hz, 4H), 3.68 - 3.58 (m, 8H ), 2.67 (t, J=6.5 Hz, 2H).

[0375] Example 12. 3-(2-(2-(2-(5-chloro-1,1-dioxide-3-oxo Isothiazole-2(3H)-yl)ethoxy)ethoxy)ethoxy)propanoic acid [ka] In DCM (30 mL), 3-[2-[2-[2-(5-chloro-1,1,3-triox So-isothiazole-2-yl)ethoxy]ethoxy]ethoxy]propanebenzyl acid ( A solution of 560 mg (1.21 mmol, 1 equivalent) is mixed with methanesulfonic acid (1.17 g, 1 2.12 mmol (863.06 μL, 10 equivalents) was added dropwise at 10°C. This mixture The solution was heated to 35°C and stirred for 10 hours. The solution turned yellow. The residue was then mixed with water (30 ml). * 3) The mixture was washed, and then the organic phase was dried with anhydrous Na2SO4, filtered, and vacuum concentrated. Residue separation TLC (ethyl acetate:ethyl acetate:methanol:acetic acid = 40:8:1, Rf = Purified by 0.77), 3-[2-[2-[2-(5-chloro-1,1,3-trioxyl So-isothiazole-2-yl)ethoxy]ethoxy]ethoxy]propanoic acid (95.6 1 mg, 240.16 μmol, yield 19.81%, purity 93.389%) as a colorless oil. Obtained as a physical substance; LC-MS (ES, m / z): 372.1 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ = 12 .13 (br s, 1H), 7.64 (s, 1H), 3.86 - 3.74 (m, 2H), 3.61 (td, J=6.0, 16.9 Hz, 4H) , 3.54 - 3.46 (m, 8H), 2.43 (t, J=6.4 Hz, 2H).

[0376] I.34. 3-(2-(2-(2-(4,5-dichloro-3-oxoisothiazole- 2(3H)-Iyl)ethoxy)ethoxy)ethoxy)benzyl propanoate [ka] In DCM (30 mL), 3-[2-[2-[2-(5-chloro-3-oxo-isothia Benzyl zole-2-yl ethoxyethoxyethoxypropanoate (1.6g, 3 To a solution of 0.72 mmol, 1 equivalent, sulfuryl chloride (1.00 g, 7.44 mmol, 744.17 μL (2 equivalents) was added dropwise at 0°C. The mixture was stirred at 0-10°C for 12 hours. A clear, pale yellow solution was obtained after the addition of sulfuryl chloride. TLC (ethyl acetate: stone) The oil-ether ratio (2:1, Rf = 0.5) indicated that the reaction was complete. The mixture was vacuum concentrated to obtain the crude product. The residue was poured into H2O (50 mL) and DCM (50 mL) * Extracted in step 2). The combined organic layers were washed with H2O (50 mL) and dried with Na2SO4. The residue was filtered and vacuum concentrated. Column chromatography of the residue was performed using silica gel (ethyl acetate). Refined by ru: petroleum ether (1:2~2:1), 3-[2-[2-[2-(4,5- Dichloro-3-oxo-isothiazole-2-yl)ethoxy]ethoxy]ethoxy]p Benzyl ropanoate (1.06g, 1.76 mmol, yield 47.17%, purity 76.9%) ) was obtained as a colorless oily substance; 1 ¹H NMR (400 MHz, chloroform-d) δ ppm 7.28 - 7.40 (m, 4 H) 5.13 (s, 2 H) 3.98 - 4.04 (m, 2 H) 3.77 (t, J=6.39 Hz, 2 H) 3.67 - 3.72 (m, 2 H) 3.57 - 3.67 (m, 8 H) 2.65 (t, J=6.39 Hz, 2 H).

[0377] I.35. 3-(2-(2-(2-(4,5-dichloro-1,1-dioxide-3-o Xisoisothiazole-2(3H)-yl)ethoxy)ethoxy)ethoxy)propanoate Njil [ka] In H2O (20 mL), CH3CN (10 mL), and DCM (10 mL), 3-[2 -[2-[2-(4,5-dichloro-3-oxoisothiazol-2-yl)ethoxy ]Ethoxy]ethoxy]benzyl propanoate (1g, 2.15 mmol, 1 equivalent) and A mixture of NaIO4 (1.84 g, 8.61 mmol, 477.32 μL, 4 equivalents) RuCl3.H2O (7.28 mg, 32.30 μmol, 0.015 equivalents) under N2, The mixture was added at 0°C. This mixture was stirred at 0-10°C for 2 hours. TLC showed that the reaction was complete. This demonstrated that the mixture was diluted with dimethyl (50 mL) and filtered to remove insoluble solids. Removed. The organic layer was separated and vacuum concentrated. The residue was separated into TLC (petroleum ether:ethyl acetate). Purified by (=1:1, Rf=0.6), 3-[2-[2-[2-(4,5-dichloro- 1,1,3-Trioxo-isothiazole-2-yl)ethoxy]ethoxy]ethoxy] Benzyl propanoate (830 mg, 1.61 mmol, yield 74.96%, purity 96.5%) 33% was obtained as a colorless oily substance; 1 ¹H NMR (400 MHz, chloroform-d) δ ppm 7.28 - 7.41 (m, 5 H) 5.15 (s, 2 H) 3.91 - 3.97 (m, 2 H) 3.75 - 3.83 (m, 4 H) 3.59 - 3. 68 (m, 8 H) 2.66 (t, J=6.50 Hz, 2 H).

[0378] Example 13. 3-(2-(2-(2-(4,5-dichloro-1,1-dioxide-3- Oxoisothiazole-2(3H)-yl)ethoxy)ethoxy)ethoxy)propanoic acid [ka] In DCM (10 mL), 3-[2-[2-[2-(4,5-dichloro-1,1,3- Lioxo-isothiazole-2-yl)ethoxy]ethoxy]ethoxy]propanoate ben In a solution of zir (820.00 mg, 1.65 mmol, 1 equivalent), methanesulfonic acid (1 (0.59 g, 16.52 mmol, 1.18 mL, 10 equivalents) was added dropwise at 10°C. Next, This solution was heated to 35°C and stirred for 10 hours. The residue was diluted with DCM (20 ml), and then... Then, add this solution to water (15 ml) * 3) Wash, and dry the organic phase with anhydrous Na2SO4. The solution was filtered and vacuum concentrated. The residue was separated by TLC (ethyl acetate:acetic acid = 250:1, Rf = 0). Purified by 55), 3-[2-[2-[2-(4,5-dichloro-1,1,3-trio Xo-isothiazol-2-yl)ethoxy]ethoxy]ethoxy]propanoic acid (249 0.4 mg, 602.76 μmol, yield 36.49%, purity 98.180%) as a yellow oil. Obtained as a physical substance; LC-MS (ES, m / z): 406.0 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ = 12.16 (br s, 1H), 3.85 (t, J=5.5 Hz, 2H), 3.65 (br t, J=5.4 Hz, 2H), 3.61 - 3.45 (m, 10H), 2.43 (t, J=6.3 Hz, 2H).

[0379] I.36. 1-Amino-3,6,9,12,15,18-Hexaoxahenicosan-2 Benzyl 1-Oate 4-methylbenzenesulfonate [ka] In toluene (30 mL), phenylmethanol (2.45 g, 22.64 mmol, 2 0.35 mL, 8 equivalents), 3-[2-[2-[2-[2-[2-(2-aminoethoxy) Toxyethoxyethoxyethoxyethoxypropanoic acid (1g, 2.83mO l, 1 equivalent) and TsOH.H2O (565.16 mg, 2.97 mmol, 1.05 The mixture (in equivalent amounts) was stirred at 140°C for 14 hours using a Dean-Stark trap. The mixture changed from turbid to clear after several hours. The residue was vacuum concentrated to remove toluene. Next, TBME (50 ml) was poured into the residue and stirred for 1 minute. Then, the supernatant was removed and the dry Dry and 3-[2-[2-[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy Benzyl ethoxyethoxyethoxypropanoate; 4-methylbenzene sulfone The acid (1.45 g, crude product) was obtained as a yellow oily substance; 1 H NMR (400 MHz, chloroform-d) ) δ = 7.80 (d, J=8.1 Hz, 2H), 7.67 - 7.46 (m, 2H), 7.39 - 7.31 (m, 5H), 7.15 (d , J=7.8 Hz, 2H), 5.13 (s, 2H), 3.96 - 3.83 (m, 2H), 3.75 - 3.50 (m, 22H), 3.24 - 3.14 (m, 2H), 2.63 (t, J=6.2 Hz, 2H), 2.34 (s, 3H).

[0380] I.37. 23,30-Dioxo-4,7,10,13,16,19,34,37,4 0,43,46,49-dodecaoxa-26,27-dithia-22,31-diazadopene Dibenzyl tacontanedioate [ka] In DCM (5 mL), 3-(2-carboxyethyl disulfanyl)propanoic acid (47 (0.41 mg, 225.47 μmol, 1 equivalent) and TEA (91.26 mg, 901. 86 μmol, 125.53 μL, 4 equivalents), HOBt (91.40 mg, 676.40 μmol, 3 eq), EDCI (129.67 mg, 676.40 μmol, 3 eq) The mixture contains 3-[2-[2-[2-[2-[2-(2-aminoethoxy)ethoxy]eth [Xy]ethoxy]ethoxy]ethoxy]benzyl propanoate; 4-methylbenzene sulfonate Acid (277.65 mg, 450.93 μmol, 2 equivalents) was added dropwise at 25°C. The mixture was stirred at 25°C for 8 hours. TLC (ethyl acetate:methanol = 3:1, R f=0.33) indicated that the starting materials were consumed and new major spots had emerged. The residue was poured into saturated NaCl (10 ml) and stirred for 2 minutes. Next, the aqueous phase was converted to DCM (5 ml). L * 3) Extracted. Combined organic phase with saturated NaCl (10 mL) * 2) Wash and anhydrous N The residue was dried with a2SO4, filtered, and vacuum concentrated. The residue was separated by preparative HPLC (column: Water). RS Xbridge 150 * 25 5μ; Mobile phase: [Water (10mM NH4HCO3 )-ACN];B%:32%~62%, 12 min) Purified by 3-[2-[2-[2- [2-[2-[2-[3-[[3-[2-[2-[2-[2-[2-[2-(3-Benji Luoxy-3-oxopropoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy [C]ethylamino]-3-oxopropyl]disulfanyl]propanoylamino]e Toxy]Ethoxy]Ethoxy]Ethoxy]Ethoxy]Ethoxy]Benzyl propanoate (7 0 mg, 65.96 μmol, yield 29.25% was obtained as a colorless oily substance; 1 1H NMR ( 400MHz, chloroform (d) δ = 7.41 - 7.29 (m, 10H), 6.50 (br s, 2H), 5.14 (s, 4H) , 3.78 (t, J=6.5 Hz, 4H), 3.67 - 3.61 (m, 40H), 3.59 - 3.55 (m, 4H), 3.45 (q, J= 5.1 Hz, 4H), 2.97 (t, J=7.2 Hz, 4H), 2.66 (t, J=6.5 Hz, 4H), 2.60 (t, J=7.1 Hz, 4H).

[0381] I.38. 1-(5-chloro-3-oxoisothiazole-2(3H)-yl)-3, 6,9,12,15,18-Benzyl hexaoxahenicosan-21-ate [ka] 3-[2-[2-[2-[2-[2-[2-[3-[[3-[ 2-[2-[2-[2-[2-[2-(3-benzyloxy-3-oxopropoxy) Ethoxy]Ethoxy]Ethoxy]Ethoxy]Ethoxy]Ethoxy]Ethylamino]-3-oxopropyl [Ropil] disulfanyl] propanoylamino] ethoxy] ethoxy] ethoxy] ethoxy [C]ethoxy]ethoxy]-benzyl propanoate (5.5g, 5.18mmol, 1 equivalent) Add sulfuryl chloride (3.50 g, 25.91 mmol, 2.59 mL, 5 equivalents) to the solution of ). ) was added dropwise at 0°C. The mixture was stirred at 0-20°C for 12 hours. TLC (ethyl acetate) A methanol-to-methanol ratio of 10:1 with Rf values ​​of 0.3 and 0.5 indicated that the reaction was complete. Pour this mixture into ice / water (10 mL) and add DCM (20 mL) * Extracted in step 2). Combined Organic solvent H2O (20 mL) * 2) Wash with brine (20 mL) and dry with Na2SO4. The sample was dried, filtered, and vacuum concentrated. The residue was subjected to column chromatography using silica gel. The oil ether:ethyl acetate (1:1 to 0:1) is purified, and 3-[2-[2-[2-[2 -[2-[2-(3-oxoisothiazol-2-yl)ethoxy]ethoxy]ethoxy ]Ethoxy]Ethoxy]Ethoxy]Benzyl propanoate (1.4g, 2.29mmol, A brown oily substance was obtained with a yield of 22.05% and a purity of 86.148%. 1 H NMR (400 MHz, Loloform-d) δ ppm 8.02 (d, J=6.17 Hz, 1 H) 7.22 - 7.32 (m, 5 H) 6.17 (br d, J =6.17 Hz, 1 H) 5.07 (s, 2 H) 3.92 (br t, J=4.30 Hz, 2 H) 3.51 - 3.73 (m, 24 H) 2 .58 (t, J=6.39 Hz, 2 H) and 3-[2-[2-[2-[2-[2-[2-(5-Cro Ro-3-oxo-isothiazole-2-yl)ethoxy]ethoxy]ethoxy]ethoxy ]Ethoxy]Ethoxy]benzyl propanoate (3.1g, 4.44mmol, yield 42. 85% with a purity of 80.532% was obtained as a colorless oily substance; 1 ¹H NMR (400 MHz, chlorophos) Lum-d) δ ppm 7.30 - 7.40 (m, 5 H) 6.26 (s, 1 H) 5.15 (s, 2 H) 3.93 - 3.99 (m, 2 H) 3.78 (t, J=6.50 Hz, 2 H) 3.60 - 3.72 (m, 23 H) 2.66 (t, J=6.50 Hz, 2 H).

[0382] I.39 1-(5-chloro-1,1-dioxide-3-oxoisothiazole-2(3 H)-yl)-3,6,9,12,15,18-hexaoxahenicosan-21-ate ben Jill [ka] 3-[2-[2- [2-[2-[2-[2-(5-chloro-3-oxoisothiazol-2-yl)eth [Xy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]benzyl propanoate (1g (1.78 mmol, 1 equivalent) and NaIO4 (1.52 g, 7.12 mmol, 39 In a mixture of 4.34 μL (4 equivalents), add RuCl3.H2O (20.05 mg, 88.96 (μmol, 0.05 equivalent) was added all at once at 20°C under N2 conditions. Next, this mixture was heated for 20 The mixture was stirred at °C for 1 hour. The residue was then mixed with ethyl acetate (20 mL). * 2) Extracted. Combined organic phase The residue was dried with anhydrous Na2SO4, filtered, and vacuum concentrated. Column chromatography was performed on the residue. (SiO2, petroleum ether:ethyl acetate = 1:1 ~ ethyl acetate:methanol = 10:1) Purified by 3-[2-[2-[2-[2-[2-[2-(5-chloro-1,1,3- Trioxo-isothiazole-2-yl)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] [Toxy]ethoxy]propanoate benzyl (520 mg, 849.06 μmol, yield 47 A yellow oily substance with a purity of 97% and a concentration of 0.72% was obtained. 1 1H NMR (400 MHz, chloroform-d) δ = 7.44 - 7.29 (m, 5H), 6.72 (s, 1H), 5.14 (s, 2H), 3.92 - 3.86 (m, 2H), 3.80 - 3.73 (m, 4H), 3.69 - 3.59 (m, 20H), 2.66 (t, J=6.4 Hz, 2H).

[0383] Example 14. 1-(5-chloro-1,1-dioxide-3-oxoisothiazole-2) (3H)-yl)-3,6,9,12,15,18-hexaoxahenicosan-21-acid [ka] In DCM (30 mL), 3-[2-[2-[2-[2-[2-[2-(5-chloro-1 ,1,3-trioxo-isothiazole-2-yl)ethoxy]ethoxy]ethoxy]e Toxyethoxyethoxybenzylpropanoate (520 mg, 875.32 μmol) In a solution of 1 equivalent, methanesulfonic acid (841.23 mg, 8.75 mmol, 623 mg) 0.13 μL (10 equivalents) was added dropwise at 10°C. The solution was heated to 35°C and stirred for 10 hours. Residual water (30ml) * 3) Wash, then dry the organic phase with anhydrous Na2SO4. Filtered and vacuum concentrated. Separation of residue TLC (ethyl acetate:methanol:acetic acid = 40:8: 1. Purified by Rf=0.58. The residue was separated by preparative HPLC (column: Nano-mic). ro Kromasil C18 100 * 30mm 5μm; Mobile phase: [water (0.05 Purify by %HCl)-ACN];B%:1%~30%, 10 min), and 3-[2-[2- [2-[2-[2-[2-(5-chloro-1,1,3-trioxoisothiazole-2 -yl)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid ( 55.93 mg, 106.24 μmol, yield 12.14%, purity 95.726%) Obtained as a colored oily substance; LC-MS (ES, m / z): 504.2 [M+H] + ; 1 1H NMR (400MHz, DMSO-d6) δ = 12.30 - 11.96 (m, 1H), 7.64 (s, 1H), 3.83 - 3.76 (m, 2H), 3.65 - 3.58 (m, 4H) , 3.54 - 3.52 (m, 2H), 3.52 - 3.48 (m, 18H), 2.44 - 2.42 (m, 2H).

[0384] I.40 3-[2-[2-[2-[2-[2-[2-(4,5-dichloro-3-oxo -Isothiazole-2-yl)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] [Toxy]benzyl propanoate [ka] In DCM (30 mL), 3-[2-[2-[2-[2-[2-[2-(5-chloro-3 -Oxo-isothiazole-2-yl)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]eth In a solution of xy]ethoxy]benzyl propanoate (1.3g, 2.31 mmol, 1 equivalent) Sulfuryl chloride (624.34 mg, 4.63 mmol, 462.47 μL, 2 equivalents) The solution was added dropwise at 20°C. The solution was stirred at 20°C for 2 hours. The solution turned yellow. The residue... Pour into ice water (30 ml) and stir for 30 minutes. Add the DCM phase to water (50 ml) * 6) Wash, The solution was dried over anhydrous Na2SO4, filtered, and vacuum concentrated. The residue was separated by TLC (ethyl acetate). Purified by methanol (10:1, Rf=0.50), 3-[2-[2-[2-[2- [2-[2-(4,5-dichloro-3-oxoisothiazol-2-yl)ethoxy] Ethoxy]Ethoxy]Ethoxy]Ethoxy]Ethoxy]Benzyl propanoate (800mg) 1.21 mmol was obtained as a yellow oily substance with a yield of 52.19% and a purity of 90%. 1 1H NMR (400MHz, chloroform-d) δ = 7.40 - 7.29 (m, 5H), 5.15 (s, 2H), 4.04 (t, J=4.7 Hz, 2H), 3.78 (t, J=6.4 Hz, 2H), 3.72 (t, J=4.7 Hz, 2H), 3.69 - 3.63 (m, 16H), 3.62 (s, 4H), 2.66 (t, J=6.5 Hz, 2H).

[0385] I.41 1-(4,5-dichloro-1,1-dioxide-3-oxoisothiazole- 2(3H)-yl)-3,6,9,12,15,18-hexaoxahenicosan-21- benzyl acid [ka] 3-[2-[2- [2-[2-[2-[2-(4,5-dichloro-3-oxoisothiazol-2-yl )Ethoxy]Ethoxy]Ethoxy]Ethoxy]Ethoxy]Ethoxy]Benzyl propanoate (600 mg, 1.01 mmol, 1 equivalent) and NaIO4 (860.56 mg, 4. A mixture of 0.2 mmol, 222.94 μL, and 4 equivalents of RuCl3.H2O (11.3 4 mg (50.29 μmol, 0.05 equivalents) was added all at once at 0°C under N2 conditions. The mixture was stirred at 0°C for 2 minutes, then heated to 25°C and stirred for 1 hour. The residue was ethyl acetate. Pour into (30 ml) and then filter. The filtrate was then mixed with ethyl acetate (30 ml). * Extracted in 3) The combined organic phase was vacuum concentrated. The residue was separated by TLC (ethyl acetate, Rf=0.50). Further purification, 3-[2-[2-[2-[2-[2-[2-(4,5-dichloro-1,1, 3-Trioxo-isothiazole-2-yl)ethoxy]ethoxy]ethoxy]ethoxy ]Ethoxy]Ethoxy]benzyl propanoate (220 mg, 350.03 μmol, yield A colorless oily substance with a purity of 100% was obtained, containing 34.80% of the original material. 1 1H NMR (400MHz, chloroform m-d) δ = 7.40 - 7.27 (m, 4H), 5.15 (s, 2H), 3.99 - 3.90 (m, 2H), 3.78 (t, J=6. 2 Hz, 4H), 3.70 - 3.58 (m, 20H), 2.66 (t, J=6.4 Hz, 2H).

[0386] Example 15. 1-(4,5-dichloro-1,1-dioxide-3-oxoisothiazo Ru-2(3H)-yl)-3,6,9,12,15,18-hexaoxahenicosan-2 1- Acid [ka] In DCM (5 mL), 3-[2-[2-[2-[2-[2-[2-(4,5-dichloro -1,1,3-trioxoisothiazol-2-yl)ethoxy]ethoxy]ethoxy Ethoxy Benzyl Propanate (220 mg, 350.03 μm In a solution of (1 ol, 1 equivalent), methanesulfonic acid (504.60 mg, 5.25 mmol, 3) 73.78 μL (15 equivalents) was added dropwise at 10°C. Next, this solution was heated to 40°C, and 2 The mixture was stirred for 0 hours. The residue was diluted with DCM (20 ml), and then the solution was diluted with water (15 ml). * 3 The organic phase was washed, dried with anhydrous Na2SO4, filtered, and vacuum concentrated. The residue was separated. HPLC (Column: Nano-micro Kromasil C18 100) * 30m m 5μm; Mobile phase: [Water (0.05%HCl)-ACN]; B%: 25%~55%, 1 Purified by (0 min), 3-[2-[2-[2-[2-[2-[2-(4,5-dichloro- 1,1,3-Trioxo-isothiazole-2-yl)ethoxy]ethoxy]ethoxy] Ethoxy]ethoxy]ethoxy]propanoic acid (53.79 mg, 98.63 μmol, A yellow oily substance with a concentration of 28.18% and a purity of 98.724% was obtained; LC-MS (ES, m / z): 538 .2 [M+H] +; 1H NMR (400MHz, DMSO-d6) δ = 12.13 (br s, 1H), 3.89 - 3.81 (m, 2H), 3.65 (t, J=5.5 Hz, 2H), 3.59 (t, J=6.4 Hz, 2H), 3.56 - 3.48 (m, 20H), 2.43 (t, J = 6.4 Hz, 2H).

[0387] Example 16. [ka] In a flask, under argon, the product 6-(5-chloro-1,1-dioxide-3-oxide) Soisothiazole-2(3H)-yl)hexanoate (12.58 mg, 0.045 mmol) l) Add DCM (2 mL) and DMF (10 μl). Cool this mixture to 0°C. Next, oxalyl dichloride (11.65 μl, 0.136 mmol) was added dropwise. Warm the mixture to room temperature, add dry MeOH to the aliquots in LCMS (to extract the methyl ester). The mixture was stirred until complete transformation was observed (tracked by LC-MS). The crude product was evaporated under vacuum. The residue was collected in a DCM and dried again under vacuum to obtain a yellow solid. This crude material was obtained. It was used in the next step without further purification.

[0388] 2. Synthesis of drug linker complexes Example A.((2R,3R)-3-((S)-1-((3R,4S,5S)-4-((S )-2-((S)-2-((4-(6-(5-chloro-1,1-dioxide-3-oxo Isothiazole-2(3H)-yl)-N-methylhexaneamide)phenethyl)(meth (L)amino)-3-methylbutanamide)-N,3-dimethylbutanamide)-3-methylbutanamide) Xy-5-methylheptanoyl)pyrrolidine-2-yl)-3-methoxy-2-methyl (Ropanoyl)-L-phenylalanine [ka]

[0389] Standard procedure for drug-linker synthesis: In a flask, under nitrogen at room temperature, 6-(5-chloro-1,1-dioxide-3-oxo Sothiazol-2(3H)-yl)hexanoate (205 mg, 0.73 mmol) (Implemented) Example 1) Dichloromethane (10 mL) and DMF (100 μl) were added. This mixture The mixture was cooled to 0°C using an ice bath, and then oxalyl chloride was added (190, 4 μl, 2 (18 mmol). This mixture was warmed to room temperature and stirred for 2 hours. This reaction mixture was then vacuumed. It was evaporated below. The residue was taken to CH2Cl2 and dried again under vacuum, and 6-(5-chloro) was added. -1,1-dioxide-3-oxoisothiazole-2(3H)-yl)hexanoyl Lorid was obtained as a yellow solid. It was stored in a vial under N2 at room temperature (S)-2-((2R,3 R)-3-((S)-1-((3R,4S,5S)-4-((S)-N,3-dimethyl- 2-((S)-3-methyl-2-(methyl(4-(methylamino)phenethyl)amino) Butanamide)Butanamide)-3-methoxy-5-methylheptanoyl)pyrrolidine- 2-yl)-3-methoxy-2-methylpropanamide)-3-phenylpropanoic acid, ( S)-2-((2R,3R)-3-((S)-1-((3R,4S,5S)-4-((S )-N,3-dimethyl-2-((S)-3-methyl-2-(methyl(4-(methylamino )Phenethyl)amino)butanamide)butanamide)-3-methoxy-5-methylhept Tanoyl)pyrrolidine-2-yl)-3-methoxy-2-methylpropanamide)-3- Phenylpropanoic acid compound with 2,2,2-trifluoroacetic acid (1:1) (111 mg, 0 It was introduced together with (102 mmol) and dichloromethane (3.7 mL). The mixture was cooled to 0°C, and DIPEA (70.9 μl, 0.406 mmol) was added. Stir the mixture at 0°C for 10 minutes, then add 6-(5-chloro-1,1-dioxide-3- Oxoisothiazol-2(3H)-yl)hexanoyl chloride (36,6 mg, 0, Add 122 mmol) to the solution in DCM (2 mL of DCM, 248 mg of acid chloride) dropwise. The mixture was stirred at 0°C for 1 hour and 15 minutes. Trifluorovinegar was added to the mixture at 0°C. Acid (32.9 μl, 0.426 mmol), acetonitrile (2.1 mL), and water (0 The reaction was stopped by adding 3 mL of [amount missing]. The crude material was vacuum concentrated, and the residue was separated. LC (Column X-Bridge C18 (100) * 30) As a result, ACN as the mobile phase Purification was performed using a gradient of priming water (containing 0.1% TFA), and ((2R,3R)-3-((S) -1-((3R,4S,5S)-4-((S)-2-((S)-2-((4-(6-(5) -Chloro-1,1-dioxide-3-oxoisothiazole-2(3H)-yl)-N- Methylhexanamide) Phenethyl) (Methyl)amino)- 3-methylbutanamide)- N,3-dimethylbutanamide)-3-methoxy-5-methylheptanoyl)pyrrolidine (-2-yl)-3-methoxy-2-methylpropanoyl)-L-phenylalanine was obtained. The mass spectrum and 1 The H-NMR spectra are shown in the figure. This is represented in 1A and 1B.

[0390] Example B.((2R,3R)-3-((S)-1-((3R,4S,5S)-4-((S )-2-((S)-2-((4-(6-(4,5-Dichloro-1,1-dioxide-3- Oxoisothiazole-2(3H)-yl)-N-methylhexaneamide)phenethyl) (methyl)amino)-3-methylbutanamide)-N,3-dimethylbutanamide)-3 -Methoxy-5-methylheptanoyl)pyrroridine-2-yl)-3-methoxy-2-methyl (Tylpropanoyl)-L-phenylalanine [ka] This is because the starting material is 6-(4,5-dichloro-1,1-dioxide-3-oxoi Sothiazol-2(3H)-yl)hexanoic acid (Example 2) and ((2R,3R)-3 -((S)-1-((3R,4S,5S)-4-((S)-N,3-dimethyl-2-(( S)-3-methyl-2-(methyl(4-(methylamino)phenethyl)amino)butane (mid)butanamide)-3-methoxy-5-methylheptanoyl)pyrrolidine-2-yl )-3-methoxy-2-methylpropanoyl)-L-phenylalanine compound 2,2, Used with 2-trifluoroacetic acid (1:1), it is a standard technique for drug-linker synthesis. They were synthesized in order.

[0391] The mass spectrum of this drug-linker complex is shown in Figure 2.

[0392] Example C.((2R,3R)-3-((S)-1-((3R,4S,5S)-4-((S )-2-((S)-2-((4-((((4-((S)-2-((S)-2-(6-(5 -Chloro-1,1-dioxide-3-oxoisothiazole-2(3H)-yl)hexa (Irradiate)-3-methylbutanamide)-5-ureidopentanamide)benzyl) (C)carbonyl)(methyl)amino)phenethyl)(methyl)amino)-3-methylbut (N,3-dimethylbutanamide)-3-methoxy-5-methylheptanoyl (Pyrrolidine-2-yl)-3-methoxy-2-methylpropanoyl)-L-phenyl Lanin [ka] This uses 6-(5-chloro-1,1-dioxide-3-oxoisothia as a starting material. Zole-2(3H)-yl)hexanoic acid (Example 1) and ((2R,3R)-3-(( S)-1-((3R,4S,5S)-4-((S)-2-((S)-2-((4-((( (4-((S)-2-((S)-2-amino-3-methylbutanamide)-5-ureido Pentanamide)benzyl)oxy)carbonyl)(methyl)amino)phenethyl)(Me (Cyl)amino)-3-methylbutanamide)-N,3-dimethylbutanamide)-3-methylbutanamide)-3-methylbutanamide) Toxy-5-methylheptanoyl)pyrroridine-2-yl)-3-methoxy-2-methyl Standard procedure for drug-linker synthesis using propanoyl-L-phenylalanine It was obtained according to this.

[0393] The mass spectrum of this drug linker complex and 1 The H-NMR spectra are shown in the figure. This is represented in 3A and 3B.

[0394] Example D.((2R,3R)-3-((S)-1-((3R,4S,5S)-4-((S )-2-((S)-2-((4-((((4-((S)-2-((S)-2-(6-(4 ,5-Dichloro-1,1-dioxide-3-oxoisothiazole-2(3H)-yl) Hexaneamide-3-methylbutanamide-5-ureidopentanamide-benzyl )Oxy)carbonyl)(methyl)amino)phenethyl)(methyl)amino)-3-methyl (Rubutanamide)-N,3-dimethylbutanamide)-3-methoxy-5-methylhepta (Noyl)pyrrolidine-2-yl)-3-methoxy-2-methylpropanoyl)-L-Fe Nylalanine [ka] This is because the starting material is 6-(4,5-dichloro-1,1-dioxide-3-oxoi Sothiazol-2(3H)-yl)hexanoic acid (Example 2) and ((2R,3R)-3 -((S)-1-((3R,4S,5S)-4-((S)-2-((S)-2-((4- ((((4-((S)-2-((S)-2-amino-3-methylbutanamide)-5-u Raidopentanamide)benzyl)oxy)carbonyl)(methyl)amino)phenethyl (methyl)amino)-3-methylbutanamide)-N,3-dimethylbutanamide)- 3-Methoxy-5-methylheptanoyl)pyrrolidine-2-yl)-3-Methoxy-2- Using methylpropanoyl-L-phenylalanine, a standard for drug-linker synthesis. It was obtained by following the procedure.

[0395] The mass spectrum of this drug linker complex and 1 The H-NMR spectra are shown in the figure. This is shown in 4A and 4B.

[0396] Example E.((2R,3R)-3-((S)-1-((3R,4S,5S)-4-((S )-2-((S)-2-((4-((((4-((S)-2-((S)-2-(3-(2 -(2-(2-(5-chloro-1,1-dioxide-3-oxoisothiazole-2(3 H)-Iyl)ethoxy)ethoxy)ethoxy)propanamide)-3-methylbutaneamide (D)-5-ureidopentanamide)benzyl)oxy)carbonyl)(methyl)amino )Phenethyl)(methyl)amino)-3-methylbutanamide)-N,3-dimethylbutanamide (Hydramide)-3-methoxy-5-methylheptanoyl)pyrrolidine-2-yl)-3-methyl Toxy-2-methylpropanoyl)-L-phenylalanine 2,2,2-trifluorovinegar Salt salts [ka] As a starting material, 3-(2-(2-(2-(5-chloro-1,1-dioxide-3-oxy Soisothiazole-2(3H)-yl)ethoxy)ethoxy)ethoxy)propanoic acid (actual Example 12) and (S)-2-((2R,3R)-3-((S)-1-((3R,4S, 5S)-4-((S)-2-((S)-2-((4-((((4-((S)-2-((S )-2-amino-3-methylbutanamide)-5 ureidopentanamide) benzyl) Xy(carbonyl)(methyl)amino)phenethyl)(methyl)amino)-3-methyl Tanamide)-N,3-dimethylbutanamide)-3-methoxy-5-methylheptanoide (L)pyrroridine-2-yl)-3-methoxy-2-methylpropanamide)-3-phenyl The drug-linker was synthesized using lupropanoic acid, following a standard procedure for drug-linker synthesis.

[0397] The mass spectrum of this drug linker complex and 1 The H-NMR spectra are shown in the figure. This is shown in 5A and 5B.

[0398] Example F. ((2R,3R)-3-(1-((3R,4R,5S)-4-((S)-2- ((S)-2-((4-((S)-2-((S)-2-(3-(2-(2-(2-(5- Chloro-1,1-dioxide-3-oxoisothiazole-2(3H)-yl)ethoxy Ethoxy)Ethoxy)propanamide)-3-methylbutanamide)-N-methylpro Panamide)phenethyl)(methyl)amino)-3-methylbutanamide)-N,3-di Methylbutanamide)-3-methoxy-5-methylheptanoyl)pyrrolidine-2-yl )-3-methoxy-2-methylpropanoyl)-D-phenylalanine [ka] As a starting material, 3-(2-(2-(2-(5-chloro-1,1-dioxide-3-oxy Soisothiazole-2(3H)-yl)ethoxy)ethoxy)ethoxy)propanoic acid (actual Example 12) and ((2R,3R)-3-(1-((3R,4R,5S)-4-((S) -2-((S)-2-((4-((S)-2-((S)-2-amino-3-methylbutane (amide)-N-methylpropanamide)phenethyl)(methyl)amino)-3-methyl Tanamide)-N,3-dimethylbutanamide)-3-methoxy-5-methylheptanoide Pyrrolidine-2-yl-3-methoxy-2-methylpropanoyl-D-phenyl Alanine was used, and the drug-linker was synthesized according to a standard procedure for drug-linker synthesis.

[0399] The mass spectrum of this drug linker complex and 1 The H-NMR spectra are shown in the figure. This is shown in 6A and 6B.

[0400] Example G. ((2R,3R)-3-(1-((3R,4R,5S)-4-((S)-2- ((S)-2-((4-((S)-2-((S)-2-(6-(5-chloro-1,1-di Oxido-3-oxoisothiazole-2(3H)-yl)hexaneamide)-3-methyl Rubutanamide)-N-methylpropanamide)phenethyl)(methyl)amino)-3- Methylbutanamide)-N,3-dimethylbutanamide)-3-methoxy-5-methyl Butanoyl)pyrrolidine-2-yl)-3-methoxy-2-methylpropanoyl)-D- Phenylalanine [ka] As a starting material, 6-(5-chloro-1,1-dioxide-3-oxoisothiazole- 2(3H)-yl)hexanoic acid (Example 1) and ((2R,3R)-3-(1-((3 R,4R,5S)-4-((S)-2-((S)-2-((4-((S)-2-((S) (-2-amino-3-methylbutanamide)-N-methylpropanamide)phenethyl) Methyl)amino)-3-methylbutanamide)-N,3-dimethylbutanamide)-3- Methoxy-5-methylheptanoyl)pyrrolidine-2-yl)-3-methoxy-2-methyl Using lupropanoyl-D-phenylalanine, a standard method for drug-linker synthesis. It was synthesized according to the procedure.

[0401] The mass spectrum of this drug linker complex and 1 The H-NMR spectra are shown in the figure. This is shown in 7A and 7B.

[0402] Example H. (2-((2S,4S)-2,5,12-trihydroxy-7-methoxy- 4-(((1S,3R,4aS,9S,9aR,10aS)-9-methoxy-1-methyl Octahydro-1H-pyrano[4',3':4,5]oxazolo[2,3-c][1,4 ]Oxazine-3-yl)oxy)-6,11-dioxo-1,2,3,4,6,11- Hexahydrotetracene-2-carboxamide)ethyl)carbamate 4-((S)-2 -((S)-2-(6-(5-chloro-1,1-dioxide-3-oxoiisothiazole -2(3H)-yl)hexaneamide)-3-methylbutanamide)-5-ureidopen Tanamide (benzyl [ka] Example H was synthesized according to the following synthesis route. [ka] I.42.(2S,4S)-2,5,12-trihydroxy-7-methoxy-4-((( 1S,3R,4aS,9S,9aR,10aS)-9-methoxy-1-methyloctahydr Ro-1H-pyrano[4',3':4,5]oxazolo[2,3-c][1,4]oxadi 0-3-yl)oxy)-6,11-dioxo-1,2,3,4,6,11-hexahydr Rotetracene-2-carboxylic acid In a flask, add PNU-1 to a mixture of methanol (15 mL) and water (10 mL). 59682 (52 mg, 0.081 mmol) was added. NaI04 (in 5 mL of water) A solution of 34.7 mg (0.162 mmol) was added. This reaction mixture was analyzed by LC-MS. The mixture was stirred at room temperature until complete conversion was observed. The solvent was removed under vacuum to obtain I as a red solid. A result of 0.42 was obtained and used as is in the next step.

[0403] I.43((S)-1-(((S)-1-((4-((((2-aminoethyl)carbam Iyl(oxy)methyl(phenyl)amino)-1-oxo-5-ureidopentane-2- (Il)amino)-3-methyl-1-oxobutan-2-yl)carbamic acid (9H-Fur) Oren-9-yl)methyl In a flask, under argon, in DMF (0.443 mol / L), ((S)-1-((( S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-urei Dopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbami (9H-fluoren-9-yl)methyl fluoride (1g, 1.662 mmol) and carbonate Su(4-nitrophenyl) (1.011 g, 3.32 mmol) was added. Next, this mixture The mixture was cooled to 0°C, and DIPEA (639 μL, 3.66 mmol) was added dropwise. The mixture was heated to room temperature and stirred for 18 hours. This crude mixture was concentrated under vacuum. Crude product The substance was placed in a 1:1 mixture of Et2O / SiO and filtered. The precipitate was then mixed with Et2O and 5% HCl. The mixture was washed with ethanol, H2O, and then again with Et2O to obtain a yellow solid. This solid was then processed using an automated column. Chromatography silica gel (100 DCM:0 MeOH~80 DCM:20 Purified by MeOH, 345 mg of ((S)-3-methyl-1-(((S)-1- ((4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)ami (no)-1-oxo-5-ureidopentan-2-yl)amino)-1-oxobutan-2 -yl)carbamate (9H-fluoren-9-yl)methyl (white solid) was obtained, yield: 27.1%.

[0404] In DMF (6 mL), a solution of the previous product (150 mg, 0.196 mmol) was added to H OBt (34.4 mg, 0.254 mmol) and pyridine (63.3 μl, 0.78 (2 mmol) was added at 0°C. After 5 minutes, (2-) was added to this mixture in DMF (1.5 mL). tert-butyl 1-2 aminoethyl carbamate (40.7 mg, 0.254 mmol) l) Then, DIPEA (102 μl, 0.587 mmol) was added. This mixture The mixture was heated to room temperature and stirred for 2 hours. The crude product was concentrated under vacuum to obtain a white solid, which was then processed automatically. Column chromatography silica gel (100 DCM: 0 MeOH ~ 80 DCM) Purified by 20 MeOH, 129 mg of tert-butylethane-1,2-di Ludicarbamate 4-((S)-2-((S)-2-((((9H-Fluorene-9-I (Lu)Methoxy)Carbonyl)Amino)-3-Methylbutanamide)-5-Ureidopenta We obtained benzyl amide (white solid) in a yield of 84%.

[0405] In a flask, add the previous product (154 mg, 0.195 mmol) in DCM (6 mL). I added it. I cooled this mixture to 0°C and added TFA (753 μL, 9.77 mmol). This mixture was stirred at 0°C until complete conversion was observed by LC-MS. Crude mixture Vacuum concentration was performed to obtain I.43 as a white solid (quantitative yield).

[0406] I.44 (2-((2S,4S)-2,5,12-trihydroxy-7-methoxy-4 -(((1S,3R,4aS,9S,9aR,10aS)-9-methoxy-1-methylo kutahydro-1H-pyrano[4',3':4,5]oxazolo[2,3-c][1,4] Oxazine-3-yl(oxy)-6,11-dioxo-1,2,3,4,6,11-he Xahydrotetracene-2-carboxamide)ethyl)carbamate 4-((S)-2- ((S)-2-amino-3-methylbutanamide)-5-ureidopentanamide)ben Jill Flasks contain I.42 (50.9 mg, 0.081 mmol) and I.43 (78.0 mg) , 0.097 mmol) and DMF (8 mL), then HATU (30.8 mg, 0 (0.081 mmol) and DIPEA (56.7 μl, 0.324 mmol) were added. This reaction mixture was stirred at room temperature for 18 hours. Next, piperidine (80 μl) was added to this mixture. (0.811 mmol) was added. This reaction mixture was left for 1 hour (until complete conversion by LC-MS). Stirred (until visible). This mixture was concentrated under vacuum. The resulting crude product was immediately transferred to Dynamic column chromatography Silica gel (100 DCM: 0 MeOH / NH3 aqueous solution) Purified with solution ~85 DCM:25 MeOH / NH3 aqueous solution, 20 mg of I.44 (Red oily substance) was obtained, yielding 23%.

[0407] Example H. In a flask, under N2 conditions, 6-(5-) in DCM (1 mL) and DMF (10 μl) Rolo-1,1-dioxide-3-oxoisothiazole-2(3H)-yl)hexanoic acid (Example 1) (7.86 mg, 0.028 mmol) was added.

[0408] This mixture was cooled to 0°C, and then oxalyl chloride (7.28 μl, 0.085 mm) was added. (ol) was added dropwise. This mixture was heated to room temperature and LC-MS was used to observe the complete conversion. (By adding dry MeOH to the aliquot to form a methyl ester, LCMS (Tracking) The crude mixture was stirred. This crude mixture was evaporated under vacuum. The residue was taken to DCM and again Dry under vacuum to obtain 6-(5-chloro-1,1-dioxide-3-oxoisothiazole) -2(3H)-yl)hexanoyl chloride was obtained as a yellow solid (quantitative yield). The crude material was used in the next process without further refinement.

[0409] In a flask, under N2 conditions at room temperature, I.44 (20 mg, 0.0) in DCM (2 mL) 19 mmol) was introduced. This mixture was cooled to 0°C, and DIPEA (12.96 μl, (0.074 mmol) was added. This mixture was stirred at 0°C for 10 minutes, and then DCM ( Add the diluted product from the previous step (8.40 mg, 0.028 mmol) to 1 mL. Next, stir this mixture at 0°C for 2 hours (until complete conversion is observed by LC-MS). This crude mixture was concentrated under vacuum and subjected to automated column chromatography and silica gel (1 Purified using 00 DCM:0 MeOH~85 DCM:15 MeOH, 6.85 mg of Example H (also known as compound F562524) was obtained as a red solid, yielding 27%.

[0410] This drug linker complex 1The 1H-NMR spectrum is shown in Figure 8.

[0411] Example I. 4-((S)-2-((S)-2-(6-(5-chloro-1,1-dioxy Do-3-oxoisothiazole-2(3H)-yl)hexaneamide)-3-methylbut (2-Ureidopentanamide)benzyl(2-Oxo-2-((2S,4S )-2,5,12-trihydroxy-7-methoxy-4-(((1S,3R,4aS,9 S,9aR,10aS)-9-methoxy-1-methyloctahydro-1H-pyrano[4' ,3':4,5]Oxazolo[2,3-c][1,4]Oxazine-3-yl)oxy) -6,11-Dioxo-1,2,3,4,6,11-Hexahydrotetracene-2-yl Ethyl ethane-1,2-diylbis(methylcarbamate) [ka] Example I was synthesized according to the following synthesis route. [ka] I.45 (Perfluorophenyl)carbonate 2-oxo-2-((2S,4S)-2,5, 12-trihydroxy-7-methoxy-4-(((1S,3R,4aS,9S,9aR, 10aS)-9-methoxy-1-methyloctahydro-1H-pyrano[4',3':4, 5]Oxazolo[2,3-c][1,4]oxazin-3-yl)oxy)-6,11- Dioxo-1,2,3,4,6,11-hexahydrotetracene-2-yl)ethyl In a flask, under argon, PNU-159682 (12 mg, 0.0180 mmol) And DMF (1.5 mL) was added. This mixture was cooled to 0°C and bis(perfull) carbonate was added. Olophenyl (36.9 mg, 0.094 mmol) was added. Next, DMF (0.5 In a mL (ml) solution, slowly dissolve DIPEA (9.80 μl, 0.056 mmol) over 5 minutes. The mixture was then stirred at 0°C for 3 hours (conversion was confirmed by LC-MS). ). This crude mixture is concentrated under vacuum and subjected to automated column chromatography, silica gel (100). DCM:0[80 DCM:20 MeOH]~50 DCM:50[80 DCM: It was purified using [2 0 MeOH]) and 5.52 mg of I.45 was obtained as a red oily substance. rate 36%.

[0412] I.46 ((S)-3-methyl-1-(((S)-1-((4-((((4-nitro Phenoxycarbonyloxymethylphenylamino-1-oxo-5-urei Dopentan-2-yl)amino)-1-oxobutan-2-yl)carbamic acid (9H- Fluoren-9-yl)methyl 2,2,2-trifluoroacetate In a flask, under argon, in DMF (0.443 mol / L), (((S)-1-(( (S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxo-5-ure Idopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carb Methyl 9H-fluoren-9-yl (1g, 1.662 mmol) and bicarbonate Su(4-nitrophenyl) (1.011 g, 3.32 mmol) was added. Next, this mixture The mixture was cooled to 0°C, and DIPEA (639 μL, 3.66 mmol) was added dropwise. The mixture was heated to room temperature and stirred for 18 hours. This crude mixture was concentrated under vacuum to obtain Et2 The solution was taken in O / siRNA (1 / 1) and filtered. The precipitate was mixed with Et2O, 5% citric acid, and H2O. Next, it was washed again with Et2O to obtain a yellow solid. This solid was then subjected to automated column chromatography. For silica gel (100 DCM:0 MeOH~80 DCM:20 MeOH) Further purification yielded 345 mg of a white solid, yielding 27.1%. In DMF (6 mL), this In a solution of the compound (118 mg, 0,154 mmol), HOBt (27.0 mg, 0.2 (00 mmol) and pyridine (49.8 μl, 0.616 mmol) were added at 0°C. After 5 minutes, add methyl(2-(methylamino)ethyl) to this mixture in DMF (1.5 mL) tert-butyl carbamate (37.7 mg, 0.200 mmol), followed by DIP EA (81.0 μl, 0.462 mmol) was added. Next, the mixture was warmed to room temperature. The mixture was stirred for 2 hours (until complete conversion was observed by LC-MS). This crude mixture was then subjected to vacuum. The mixture was concentrated to obtain a yellow oily substance, which was then subjected to automatic column chromatography using silica gel (100). Purified using DCM:0 MeOH (~80 DCM:20 MeOH), 103 mg A white solid was obtained, with a yield of 82%.

[0413] In a flask, add the product (198 mg, 0.243 mmol) in DCM (12 mL). [Added]. This mixture was cooled to 0°C, and TFA (935 μl, 12.13 mmol) was added. This mixture was stirred at 0°C for 4 hours (until complete conversion was observed by LC-MS). This crude mixture was concentrated under vacuum to obtain 220 mg of I.46 as a clear yellow solid. (Quantitative yield).

[0414] I.47 4-((S)-2-((S)-2-amino-3-methylbutanamide)-5- Ureidopentanamide)benzyl(2-oxo-2-((2S,4S)-2,5,12 -Trihydroxy-7-methoxy-4-(((1S,3R,4aS,9S,9aR,10 aS)-9-methoxy-1-methyloctahydro-1H-pyrano[4',3':4,5] Oxazolo[2,3-c][1,4]oxazin-3-yl)oxy)-6,11-diode Xo-1,2,3,4,6,11-hexahydrotetracene-2-yl)ethyl)ethane -1,2-diirbis(methylcarbamate) In DMF (1 mL), a solution of product I.45 (19 mg, 0.022 mmol) was added to the room. At warm temperature, product 1.46 (22.2 mg, 0.027 mmol) in DMF (1 mL) Then, a solution of DIPEA (15.59 μl, 0.089 mmol) was added. This reaction mixture The mixture was stirred at room temperature for 3 hours (until complete conversion was observed by LC-MS). Next, this mixture was mixed. Piperidine (22.09 μl, 0.223 mmol) was added to the substance. This reaction mixture 1 The mixture was stirred for a certain amount of time (complete conversion was observed by LC-MS). This crude mixture was concentrated under vacuum. Automated column chromatography, silica gel (100 DCM: 0 MeOH / NH3 (9 / 1) ~75 DCM:25 MeOH / NH 3( Purified by 9 / 1)) and 10m g of I.47 was obtained as a red oily substance, yield 39%.

[0415] Example I. In a flask, under N2 conditions, 6-(5-) in DCM (1 mL) and DMF (10 μl) Rolo-1,1-dioxide-3-oxoisothiazole-2(3H)-yl)hexanoic acid (Example 1) (7.86 mg, 0.028 mmol) was added.

[0416] This mixture was cooled to 0°C, and then oxalyl chloride (7.28 μl, 0.085 mm) was added. (ol) was added dropwise. This mixture was heated to room temperature and LC-MS was used to observe the complete conversion. (By adding dry MeOH to the aliquot to form a methyl ester, LCMS (Tracking) The crude mixture was stirred. This crude mixture was evaporated under vacuum. The residue was taken to DCM and again Dry under vacuum to obtain 6-(5-chloro-1,1-dioxide-3-oxoisothiazole) -2(3H)-yl)hexanoyl chloride was obtained as a yellow solid (quantitative yield). The crude material was used in the next process without further refinement.

[0417] In a flask, under N2 conditions at room temperature, product I.47 (10 mg, 0. (0086 mmol) was introduced. This mixture was cooled to 0°C, and DIPEA (6.0 μl, (0.034 mmol) was added. This mixture was stirred at 0°C for 10 minutes, and then DCM ( The diluted previous product (3.90 mg, 0.013 mmol) was added to 1 mL of the solution. Then, this mixture was stirred at 0°C for 2 hours (until complete conversion was observed by LC-MS). Crude product is vacuum concentrated and subjected to automated column chromatography, silica gel (100 DCM: Purified using 0 MeOH (85 DCM:15 MeOH), 2.45 mg of Example I This was obtained as a red solid, with a yield of 19%.

[0418] This drug linker complex 1 The 1H-NMR spectrum is shown in Figure 9.

[0419] Example J. [ka] The synthesis was carried out according to the following synthesis route in Example J. [ka] Compound I.50 was prepared according to the following synthetic route. [ka] Compound I.48: In a flask, under argon, 2-([1,1'-biphenyl]- in DCM (5 mL) 4-yl)propan-2-ol (1g, 4.71 mmol) and pyridine (0.46 5 ml (5.75 mmol) was added. Next, this mixture was cooled to 0°C and dried DCM ( Phenylenide (0.662 ml, 5.28 mmol) was added dropwise to 2.4 mL. The reaction mixture was heated to room temperature and stirred for 18 hours (confirmed by LC-MS). finished product The mixture was vacuum concentrated. The solid mixture was dissolved in DCM and washed three times with brine. The organic layer was N Dried with a2SO4, filtered, and vacuum concentrated, yielding target compound I.48, 880 mg, 5 6% was obtained as a white solid. LCMS (ESI): 333.40 (MH+).

[0420] Compound I.49: N,N'-dimethyl-1,2-ethanediamine (2791 μl, 26.2 mmol), N-ethyl-N-isopropylpropan-2-amine (305 μl, 1.748 mmol) ) and DMF (4 mL) are placed in a flask, and under argon, in DMF (1.5 mL) , carbonate 2-([1,1'-biphenyl]-4-yl)propane-2-ylphenyl(58 A 1 mg (1.748 mmol) solution was added at 0°C. The reaction mixture was then warmed to room temperature. The mixture was stirred for 24 hours (confirmed by LC-MS). The crude product was vacuum concentrated, and the residue was collected in an automated tanker. DCM (Interchim, solid deposit): Purified using MeOH:9 / 1. The target fraction was vacuum concentrated to obtain the target compound I.49, yielding 4. 33 mg was obtained as a yellow oily substance, 76% of which was present. LCMS(ESI):327.43(MH+ ).

[0421] Compound I.50: Bis(trichloromethyl) carbonate (157 mg, 0.531 mmol) and toluene ( In a flask containing 4.3 mL of toluene (2.9 mL), methyl (2-(methylamino)ethyl)carbamate 2-([1,1'-biphenyl]-4-I (Lu) Propan-2-yl (433 mg, 1,326 mmol) and triethylamine ( 368 μl of the solution (2.65 mmol) was added at 0°C. The reaction mixture was then warmed to room temperature. The solution was stirred for 1 hour (confirmed by LC-MS). The solution was filtered, the solvent was concentrated under vacuum, and the residue was removed. Automated column chromatography (Interchim, solid deposit) The solution was purified using cyclohexane / ethyl acetate 7 / 3. The target fraction was vacuum concentrated and processed for the desired outcome. Compound I.50, yield 166 mg, 33% obtained as a white solid. LCMS(ESI):4 05.60 (MH+).

[0422] Compound I.52 was prepared according to the following synthetic route. [ka] Compound I.51: (8S,10S)-6,8,11-trihydroxy-8-(2-hydroxyacetyl) -1-Methoxy-10-(((1S,3R,4aS,9S,9aR,10aS)-9 Toxy-1-methyloctahydro-1H-pyrano[4',3':4,5]oxazolo[2 ,3-c][1,4]oxazine-3-yl)oxy)-7,8,9,10-tetrahydr Rotetracene-5,12-dione (50 mg, 0.078 mmol), 4-dimethylamine Nopyridine (47.6 mg, 0,390 mmol), molecular sieves 0.4 nm ( A flask containing 33 mg of DCM and 1 mL of DCM is subjected to argon gas. (5 mL) contains (2-((chlorocarbonyl)(methyl)amino)ethyl)(methyl) Bamic acid 2-([1,1'-biphenyl]-4-yl)propan-2-yl (91 mg, A 0.234 mmol solution was added. This mixture was stirred in the dark at 25°C for 5 days. The solution was filtered, the solvent was vacuum concentrated, and the residue was used in the next step without further purification.

[0423] Compound I.52: In an ice bath, in 1 ml of DCM, the solution of product I.51 was added to 0.5 ml of DCM. A solution of chloroacetic acid (96 μl, 1.169 mmol) was added. This solution was left at room temperature for 2 hours. The mixture was stirred. The solvent was vacuum concentrated, and the residue was subjected to automated column chromatography (Interchi). Purified using DCM / MeOH:9 / 1 (solid deposit). The solution was vacuum concentrated to obtain the target compound I.52, yielding 13 mg and 22% as a red solid. CMS(ESI): 756.76(MH+).

[0424] Compound I.53: In a flask, under argon, ((S)-1-(((S)-1-((4-(Hydroxymethyl (Tyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1 -Oxobutan-2-yl)carbamate (9H-fluoren-9-yl)methyl(25 0 mg, 0.485 mmol), bis(perfluorophenyl) carbonate (382 mg, 0. 970 mmol) and DMF (4 mL) were added. Next, this mixture was cooled to 0°C. N-ethyl-N-isopropylpropan-2-amine (127 μl, 0.727 mmol) The following was added dropwise: The reaction mixture was warmed to room temperature and stirred for 2 hours (confirmed by LC-MS). The crude product was vacuum concentrated. The crude product was then subjected to automated column chromatography (Inter). Chim, solid deposit): Purified by DCM / MeOH:9 / 1 The target fraction was vacuum concentrated to obtain the target compound I.53, yielding 281 mg, with 80% being a yellow oily substance. The result was obtained as follows: LCMS(ESI): 726.65 (MH+).

[0425] Compound I.54: In a flask, under argon, methyl(2-(methylamino)ethyl)carbamate 2- Oxo-2-((2S,4S)-2,5,12-trihydroxy-7-methoxy-4-( ((1S,3R,4aS,9S,9aR,10aS)-9-methoxy-1-methylocta Hydro-1H-pyrano[4',3':4,5]oxazolo[2,3-c][1,4]ox Sadin-3-yl)oxy)-6,11-dioxo-1,2,3,4,6,11-hexa Hydrotetracene-2-yl)ethyl (78 mg, 0.103 mmol), 1-hydrox Cibenzotriazole (27.9 mg, 0.206 mmol), N,N'-diisopropyl Add ethylamine (35.1 μl, 0.206 mmol) and DMF (2 mL). Next, cool this mixture to 0°C and ((S)-3-methyl-1-oxo-1-(((S )-1-oxo-1-((4-((((perfluorophenoxy)carbonyl)oxy) Methyl(phenyl)amino)propan-2-yl)amino)butan-2-yl)carbamyl Administer (9H-fluoren-9-yl)methyl fluoride (112 mg, 0.155 mmol) dropwise. The reaction mixture was then warmed to room temperature and stirred for 2 hours (confirmed by LC-MS). The product was vacuum concentrated. The crude product was subjected to automated column chromatography (Interchim). Purified using DCM / MeOH:9 / 1 (solid deposit). Target fraction. The mixture was vacuum concentrated to obtain the target compound I.54, yielding 77 mg and 58% as a red oily substance. CMS(ESI): 1298.0(MH+).

[0426] Compound I.55: In a flask, under argon, 4-((S)-2-((S)-2-((((9H-Fluorine Len-9-yl)methoxy)carbonyl)amino)-3-methylbutanamide)propane amide)benzyl(2-oxo-2-((2S,4S)-2,5,12-trihydroxy -7-Methoxy-4-(((1S,3R,4aS,9S,9aR,10aS)-9-Methoxy Xy-1-methyloctahydro-1H-pyrano[4',3':4,5]oxazolo[2, 3-c][1,4]oxazine-3-yl)oxy)-6,11-dioxo-1,2,3 ,4,6,11-Hexahydrotetracene-2-yl)ethyl)ethane-1,2-diyl Bis(methylcarbamate) (77.7 mg, 0.060 mmol) and DMF (2 Next, add morpholine (259 μl, 2.99 mL). Then, cool this mixture to 0°C and add morpholine (259 μl, 2.99 mL). mmol was added dropwise. The reaction mixture was warmed to room temperature and left for 2 hours (confirmed by LC-MS). The mixture was stirred. The crude product was vacuum concentrated. The crude product was subjected to automated column chromatography (I (interchim, solid deposit): DCM / MeOH:9 / 1 The target fraction was vacuum concentrated to obtain the target compound I.55, yielding 32 mg, with 50% being a red oil. I obtained it as a physical object. LCMS(ESI): 1075.80 (MH+).

[0427] Example J: In a flask, under argon at 25°C, 4-((S)-2 in dichloromethane (2 mL) -((S)-2-amino-3-methylbutanamide)propanamide)benzyl(2-O Kiso-2-((2S,4S)-2,5,12-trihydroxy-7-methoxy-4-(( (1S,3R,4aS,9S,9aR,10aS)-9-methoxy-1-methyloctahy Dro-1H-pyrano[4',3':4,5]oxazolo[2,3-c][1,4]oxa Zin-3-yl)oxy)-6,11-dioxo-1,2,3,4,6,11-hexahy Dorotetracene-2-yl)ethyl)ethane-1,2-diylbis(methylcarbamate) Salt (32 mg, 0.030 mmol, 1 equivalent) was added. This mixture was cooled to 0°C. N-ethyl-N-isopropylpropan-2-amine (20.74 μl, 0.119 ml) Add (mol). Stir this mixture at 0°C for 10 minutes, then add dichloromethane (2 The diluted Example 16 was added to (mL). Next, this mixture was incubated at 0°C for 2 hours (in LCMS). The mixture was stirred until a more complete transformation was observed. This crude product was vacuum concentrated and then automated column chromatography was performed. Mathematics (Interchim, 12g, solid deposit): DCM The compound was purified using MeOH:9 / 1. The target fraction was vacuum concentrated, and the target compound F5 was obtained. (Also known as 62646), yielded 17.4 mg, 40% of which was obtained as a red oily substance. LCMS ( ESI): 1338.41 (MH+).

[0428] The mass spectrum of this drug-linker complex is shown in Figure 21.

[0429] Example K. [ka] Example K was synthesized according to the following synthesis route. [ka] Compound I.56: In a flask, under argon, add the starting material SM1 (100 mg, 0.135 mmol) Then, 1 ml of DMF was added. This mixture was cooled to 0°C, and then 0.5 ml of DMF was added. ) contains 3-bromopropanoic acid (22.79 mg, 0.149 mmol) and 2,3,4 ,6,7,8,9,10-Octahydropyrimide[1,2-a]azepine (40.5 μl) A solution of 0.271 mmol was added dropwise. Next, this mixture was warmed to room temperature and measured by LCMS. The mixture was stirred until complete conversion was observed. This crude product was vacuum concentrated and collected in an automated column chromatography. Mathematics (Interchim, 12g, solid deposit): DCM The solution was purified using / MeOH:80 / 20. The target fraction was vacuum concentrated to obtain the target compound I.56. The yield was 108 mg, and 98% was obtained as a white solid. LCMS(ESI):811.35(M H+).

[0430] Compound I.57: In a flask, under argon, compound I.56 (87.0 mg, 0.107 mmol) Then, 2 mL of DCM was added. Next, this mixture was cooled to 0°C and N-hydroxysodium was added. Synimide (13.59 mg, 0.118 mmol) and 1-(3-dimethylaminopropyl sulfate) Add Ropil-3-ethylcarbodiimide hydrochloride (30.9 mg, 0.161 mmol). The reaction mixture was heated to room temperature and stirred for 2 hours (confirmed by LC-MS). This crude product was then prepared. It was vacuum concentrated. The crude material was used in the next process without further purification.

[0431] Compound I.58: In a flask, under argon, compound I.57 (97 mg, 0.107 mmol) and Add DCM (1 ml), then add the solution of compound I.49 and N in 1 ml of DCM. N'-diisopropylethylamine (37.3 μl, 0.214 mmol) was added dropwise. This mixture was stirred by LC-MS until complete conversion was observed. The crude product was concentrated under vacuum. It was then used in the next process without further purification.

[0432] Compound I.59: In a flask, under argon, compound I.58 (58 mg, 0.052 mmol) and DCM (1 ml) was added. This mixture was cooled to 0°C, and then dichloroacetic acid (86 μg) was added. (1.037 mmol) was added dropwise. The mixture was then warmed to room temperature and analyzed by LC-MS. The mixture was stirred until complete conversion was observed. This crude product was vacuum concentrated and then subjected to automated column chromatography. Raffy (Interchim, 12g, solid deposit): DCM / Me Purified using OH:80 / 20. The target fraction was vacuum concentrated, yielding the target compound I.59, with a yield of 3. 9 mg was obtained as a white solid, 86% of which was LC-MS (ESI): 882.6 (MH+).

[0433] Example K: In a flask, under argon at 25°C, dissolve compound I.59 in 1 mL of dichloromethane. This mixture was introduced. This mixture was cooled to 0°C and N-ethyl-N-isopropylpropane-2-A Min (15.83 μl, 0.091 mmol) was added. The mixture was stirred at 0°C for 10 minutes. Mix, then add Example 16 diluted in dichloromethane (2 mL). Next, mix The mixture was stirred at 0°C for 2 hours (until complete conversion was observed by LC-MS). This crude product The solution was vacuum concentrated and purified by preparative HPLC (HCOOH conditions), yielding 6 ml of the target sample (Example K). g, 22% was obtained as a white solid. LC-MS (ESI): 1165.37 (M + Na) + .

[0434] The mass spectrum and TOF-MS spectrum of this drug linker complex are shown in Figure 1. This is shown in 22A and 22B.

[0435] 3. Complex formation with somatostatin VI1. Somatostatin and 6-(5-chloro-1,1-dioxide-3-oxoisothia) Reaction with benzyl zole-2(3H)-yl)hexanoate [ka] 1 mg of lyophilized somatostatin (m exact =1636,72) in 4 mL baff (57.5%NaH2PO420mM, pH6.5, 40%ACN, 2.5%DM Solubilized with F) to a concentration of 153 μM (0.25 mg / mL). 33.5 mg of TC EP in 4 mL of buffer (57.5% NaH2PO4 20 mM, pH 6.5, 40%) Dissolved in ACN (2.5% DMF). Add to 300 μL of somatostatin solution (1 equivalent) 3 μL (1.1 equivalents) of TCEP solution was added. This solution was stirred at 37°C for 1 hour. Commercially available somatostatin:R t,1 (ACN): 1.57; MS ES+: M +3 / 3= 546.4, M +2 / 2 = 819.2. Reduced disulfide bonded somatostatin: R t,1 (ACN): 1.50; MS ES+: M +3 / 3 = 547.2, M +2 / 2 = 820 .3. 5 mg of 6-(5-chloro-1,1-dioxide-3-oxoisothiazole-2 (3H)-yl)benzyl(I.6)hexanoate was solubilized in 800 μL of ACN. Add 3 μL of 6-(5-chloro-1,1-dioxide-3-oxo to somatostatin solution A solution of benzyl sothiazole-2(3H)-yl)hexanoate (1.1 equivalents) was added. The solution was stirred at 37°C. t,1 (ACN): 1.66; MS ES+: M +3 / 3 = 637.6, M +2 / 2 = 956.0.

[0436] The same reaction was carried out in buffer pH 8 (57.5% NaH2PO4 20mM, pH 8, 40%) The procedure was performed in ACN (2.5% DMF).

[0437] The mass spectrum of the obtained composite is shown in Figure 10.

[0438] VI2. Somatostatin and 6-(4,5-dichloro-1,1-dioxide-3-oxo Reaction of benzyl sothiazole-2(3H)-yl)hexanoate: [ka] 1 mg of lyophilized somatostatin (m exact= 1636,72) 4 mL buff (57.5%NaH2PO420mM, pH6.5, 40%ACN, 2.5%DM Solubilized with F) to a concentration of 153 μM (0.25 mg / mL). 33.5 mg of TC EP in 4 mL of buffer (57.5% NaH2PO4 20 mM, pH 6.5, 40%) Dissolved in ACN (2.5% DMF). Add to 300 μL of somatostatin solution (1 equivalent) 3 μL (1.1 equivalents) of TCEP solution was added. This solution was stirred at 37°C for 1 hour. Commercially available somatostatin:R t,1 (ACN): 1.57; MS ES+: M +3 / 3= 546.4, M +2 / 2 = 819.2. Reduced disulfide bonded somatostatin: R t,1 (ACN): 1.50; MS ES+: M +3 / 3 = 547.2, M +2 / 2 = 820 .3. 5.4 mg of 6-(4,5-dichloro-1,1-dioxide-3-oxoisothia Benzyl(I.7) zole-2(3H)-yl)hexanoate can be dissolved in 800 μL of ACN. It was modified. 3 μL of 6-(4,5-dichloro-1,1-dioxypropyl alcohol) was added to this somatostatin solution. Benzyl hexanoate (1.1 equivalents) of 3-oxoisothiazole-2(3H)-yl The solution was added. This solution was stirred at 37°C. t,1 (in ACN):1.65;MS ES+:M +3 / 3 = 637.3, M +2 / 2 = 955.5.

[0439] The same reaction was carried out in buffer pH 8 (57.5% NaH2PO4 20mM, pH 8, 40%) The procedure was performed in ACN (2.5% DMF).

[0440] The mass spectrum of the obtained complex is shown in Figure 11.

[0441] 4. Complex formation with monoclonal antibodies 4.1. Synthesis, Purification, and Characterization of ADCs The following procedure applies to chimeric, humanized, and human IgG1 forms. IgG2, IgG For any other form such as 4, a person skilled in the art can apply this procedure using general knowledge. It should be understood that it is possible to do so.

[0442] Ab1 antibody is an anti-IGF1R IgG1 monoclonal antibody. This antibody is WO This corresponds to antibody 208F2 of 2015162291 (see Table 3 on page 36), and here, three The light chain CDR has sequences 9, 5, and 11; the three heavy chain CDRs have sequences It has sequences 7, 2 and 3; the light chain variable domain has the sequence of sequence number 18; and, The heavy chain variable domain has the sequence of sequence number 13.

[0443] The Ab2 antibody is an antimicrobial agent against bacterial proteins, specifically outer membrane protein A derived from E. coli. This is a related chimeric (IgG1) antibody, called c9G4 (Haeuw JF and Beck A. Pr). oteomics for development of immunotherapies, In Proteomics: Biomedical and Pharm aceutical Applications, Kluwer Academic Publishers, Ed. Hondermarck H., 2014, 24 3-278 pages; WO2015162291).

[0444] Antibody (1-5 mg / ml) is incubated at 37°C for 2-4 hours in 150 mM NaCl and 2 mM Partially dissolved in 10 mM borate buffer pH 8.4 containing EDTA with TCEP hydrochloride. It was reduced to . Generally, 6-20 mol equivalents of TCEP are used to target a DAR of around 4. Partial antibody reduction was confirmed by SDS-PAGE analysis under non-reducing conditions. Next, the antibody concentration was set to 150 mM NaCl, 2 mM EDTA, and 6% sucrose. Prepare the solution by adjusting the concentration to 1 mg / ml with 10 mM borate buffer at pH 8.4, and then add 10 mM DMSO4. A drug linker conjugate was added to the solution in an excess of 5 to 20 molars relative to the antibody.

[0445] Several drug linker complexes according to the present invention were conjugated to Ab1: Examples A and ADC1-B (non-cuttable linker) are shown respectively. Example B; These are ADC1-C, ADC1-D, ADC1-E, ADC1-F, and ADC1- Examples C, D, E, F, and G show G (a severable linker). .

[0446] The final DMSO concentration is set to maintain drug solubility in the aqueous medium during coupling. The solution was adjusted to 10%. The reaction was carried out at room temperature or 37°C for 1-4 hours. Excess drug was removed. Add 2.5 moles of N-acetylcysteine ​​per mole and incubate at room temperature for 1 hour. It was rapidly cooled by performing a suction.

[0447] 25 mM His, containing 150 mM NaCl and 6% sucrose, is left overnight at 4°C. After dialyzing with buffer pH 6.5, the recrosslinked antibody-drug conjugate is made public to those skilled in the art. Using the method of knowledge, commercially available chromatography columns and ultrafiltration units are used. The ADC was purified. The purified ADC was filtered and sterilized using a 0.2 μm filter and then stored at 4°C. Ta.

[0448] They were then analyzed using SDS-PAGE under reducing and non-reducing conditions to confirm drug binding. Furthermore, analytical TSK G300 is used to determine the monomer and aggregate form content. Further analysis was performed using SEC on a SWXL column. SEC chromatogram (Figure 13) The content of the aggregated form derived from this was lower than 5%, as shown in Table 9.

[0449] [Table 22]

[0450] SDS-PAGE analysis confirmed the formation of fully cross-linked antibody H2L2 (Figure 12). However, other species corresponding to partially cross-linked antibodies (H2L, H2, and HL) were also detected. These species are linked by intact intrachain crosslinks when the sample is heat-treated under reducing conditions. This is observed before implementation to ensure complete dissociation of the heavy and light chains (H and L) that have not been performed. It is important to pay attention to this.

[0451] Protein concentration is determined using a BCA assay with IgG as the standard. The DAR was determined using a TSK-butyl-NPR column for each purified ADC. Evaluated by C. It fell between 3.5 and 4.3 (Table 10). HIC profile The problem is that DAR0 is not seen in most synthesized ADCs, and DAR4 is the main peak. It became clear that a trace amount of DAR was observed. In fact, only ADC1-C and E showed trace amounts of DAR. It indicates 0. Furthermore, in ADC1-A, B, C and G, DAR3, DAR4 and DA Only R5 was observed. Except for ADC1-D, the main peak is DAR4. Second generation A Compared to DCs, these ADCs are more homogeneous, as shown in Table 10.

[0452] [Table 23]

[0453] Adcetris® (brentuximab vedotin) binds to cysteine ​​in antibodies. It is used as a reference because it uses the second-generation maleimide linker. It is the best representative example of a second-generation ADC, and the technology described in this context is It can be considered the third generation.

[0454] 4.2. ADC analysis by native mass spectrometry All chemical agents were purchased from Sigma-Aldrich: Ammonium acetate (A1 542), cesium iodide (21004), 2-propanol (I9516). IgGZ The ERO(A0-IZ1-010) enzyme was obtained from Genovis. The aqueous solution was ultrapure. It was prepared using a water system (Sartorius, Göttingen, Germany).

[0455] ADC1-A to ADC1-G were deglycosylated before the native MS experiment. This involves incubating 1 unit of IgGZERO per 1 milligram of ADC at 37°C for 30 minutes. This was done by applying a microconcentrator (Viva) to the ADC. (using spin, 10kD cutoff, Sartorius, Göttingen, Germany) Using six concentration / dilution cycles, a 150 mM ammonium acetate solution (pH 6.9) was prepared. Buffer exchange was performed. Protein concentration was measured using a NanoDrop spectrophotometer (Th Using Ermo Fisher Scientific (France), UV absorbance is measured It's decided.

[0456] Non-denatured (native) mass spectrometry of ADCs is performed using automated chip-based nanoelectrosplasia. - Devices (Triversa Nanomate, Advion, Ithaca, USA) The Q-TOF (Synapt G2) is linked together and operates in positive ion mode. The analysis was performed using a mass spectrometer (HDMS, Waters, Manchester, UK). The analysis was performed in m / z. The test was performed in the range of 1000 to 10000. The sample was then treated with 150 mM NH4OA at pH 6.9. Diluted in c and injected at 10 μM. Delayed extraction was performed using 2-propanol / water (50 / 50 In v / v, monovalent charged ions are produced by a 2 g / L solution of cesium iodide. It was done using n.

[0457] The nanoelectrospray voltage was set to 1.75kV, and the nitrogen nanoflow was set to 0.75psi. The settings were as follows: The cone voltage was set to 180 volts, and the back pressure was set to 6 millibars.

[0458] Figure 14 shows an example of an MS spectrum without deconvolution.

[0459] The DAR distribution (Figure 15) is from Mass Lynx 4.1 (Waters, Manchester). The MaxEnt (trademark) algorithm from the UK is used to determine the result after deconvolution. The software parameters were optimized for each spectrum.

[0460] The average DAR value (Figure 15) is calculated using the following formula (where j is the maximum drug loading number): Ta.

[0461]

number

[0462] [Table 24]

[0463] Figure 16 shows the DAR distribution determined from the raw spectrum after mass deconvolution. Two different ADCs, namely, ADC1-C according to the present invention, prepared from Ab1 antibody and drug linker complex C (Figure 1) 6B) and The same antibody (Ab1) and sulfomaleimide portion ( [ka] ) is the maleimide part ( [ka] A comparative drug linker complex synthesized from drug linker complex C equivalent to drug linker complex C substituted with ). This is an ADC, reference ADC Ref-A (Figure 16A). This is a comparison regarding the following:

[0464] ADC synthesized by using the classical maleimide chemistry method to link the drug and antibody. Regarding this, a non-uniform distribution was observed from DAR0 to DAR8 (Figure 16A), while this development ADC prepared using the sulfomaleimide chemical method by Akira is DAR4 75% of the samples were DAR0 / 2 and 6 / 8, indicating extreme uniformity (Figure 16B). The results are summarized in Table 12 below.

[0465] [Table 25]

[0466] 4.3. Inclusion of ADCs in four types of mammalian serum using ligand binding assays In vitro stability testing To confirm the acquisition of stability, an in vitro stability test was performed. This test was conducted as follows: The incubation of ADCs at 37°C for 14 days consists of samples 0, 3, 7 and The sample was collected on the 14th day. Next, various samples (D0, D3, D7, and D14) were taken. The total antibody concentration relative to the concentration of 4ADC was analyzed by LBA. In this case, the solution of each ADC was used in four types of serum (human, cynomolgus monkey, mouse, and rat). Prepared at 100 μg / ml and incubated at 37°C for up to 14 days. Next, D0, Aliquotes were taken from D3, D7, and D14 and stored at -80°C until administration. Total Ab To quantify ADC, the plate was thawed at room temperature while shaking, and both LBA assays were performed side by side. To put it simply, use a standard microtiter plate (MSD, Gazers) 50 μl of a 2 μg / ml anti-His antibody solution prepared by Berg (USA) in 1×PBS. Coat using [method]. After incubation at 4°C overnight, the assay plate is [processed]. 30% MSD Blocker A (MSD) at 37°C for 1 hour. The antigen with the recombinant His tag is then processed at Gaithersburg, USA. Add to assay buffer at a concentration of 2.5 μg / ml at 7°C for 1 hour. Washing step Afterward, the sample was analyzed in two replicates at a 1 / 5000 dilution and incubated at 37°C for 1 hour. On the other hand, standard ADC is loaded onto the assay plate in two replicates. The detection step is for total Ab For detection of 1 μg / ml goat anti-human Ig κ-sulfotag solution or ADC detection This is performed using one of the mouse monoclonal anti-drug antibodies labeled with a sulfotag. 3 After an incubation period of 1 hour at 7°C, the MSD Sector Imager was used. 150 containing surfactant (MSD, Gaithersberg, USA) Detection is performed using μL of 2×MSD read T buffer.

[0467] Determine the total antibody and ADC concentrations at each time point, and reduce the amount of total ADC or antibody at each time point by 100. Convert to a percentage.

[0468] Figures 17A, 17B, and 17C show data for three types of ADCs according to the present invention. ADC1-C (Figure 17B) was created by linking a drug to the Ab1 antibody using the sulfomaleimide chemistry method. ) and ADC1-E (Figure 17C) (by means of drug linker complex C or E, respectively) (Using classical maleimide chemistry, the drug is linked to the antibody in a reference ADC Ref- Comparison with B (Figure 17A). [ka] For comparison, we selected drug-linker models using the same bullet and non-cuttable linker. (Drug-linker of ADC Ref-B). This is then converted to the same antibody using the maleimide chemistry method. It was bound to. The selection of this comparison target was made by uncoupling from the antibody via the retro-Michael reaction. Therefore, it limits the "instability" of the reference ADC in serum. Thus, based on a cleavable linker Compared to our construct, this comparison shows the stability of our drug-linker. They supported making improvements even more significant.

[0469] As expected, ADC(ADC Re) was synthesized using the classical maleimide chemistry method. In fB), a decrease in ADC concentration is observed, but the sulfomaleimide chemical method according to the present invention is used The ADCs (ADC1-C and ADC1-E) generated by this process surprisingly lasted for 14 days. It is far more stable.

[0470] 4.4. In vitro cytotoxicity of ADCs The in vitro cytotoxicity of ADCs according to the present invention was evaluated. Nonspecific cytotoxicity To evaluate the compound, we also used an unrelated chimeric antibody (Ab2) called c9G4. Using the same DAR and the same drug linker complex, ADC2-C was conjugated in Example C. ADC2-E was fabricated in Example E, and ADC2-F was fabricated in Example F.

[0471] MCF-7 cells and NCI-H2122 cells were placed in a 96-well plate of complete growth medium. Seeds were sown (2500 cells / well). The following day, serial dilutions of the test ADC were added to the corresponding wells. The cells were added and incubated at 37°C for 6 days. Six days after the addition of ADC, the cell viability was measured. To confirm, we performed a Cell Titer Glo assay (PROM) on these plates. EGA was performed.

[0472] The results, expressed as survival percentages, are shown in Figures 18A and 18B. As shown, ADCs synthesized using unrelated antibodies were found in MCF-7 cells and NCI - No cytotoxic activity or moderate cytotoxic activity against both H2122 cells This was shown. In contrast, the ADCs of the present invention: ADC1-C, ADC1-E and ADC1- F dramatically reduced cell viability. Against NCI-H2122, ADC1-C, 7, 61.10 for ADC1-E and ADC1-F, respectively. -11 , 7, 16.10 -1 1 and 3,64.10 -11 M EC 50 The value is obtained, and for MCF7, ADC1 -C, ADC1-E, and ADC1-F are 1,04.10 respectively. -11 , 1.33.1 0 -11 and 7,39.10 -11 M EC 50 Values ​​were obtained, showing potent cytotoxic activity. vinegar.

[0473] 4.5. in vivo All experimental protocols were approved by Pierre Fabre's in-house animal experimentation committee. keta.

[0474] In the ovarian cancer model, 7-week-old female SCID mice (Charles RIVER Lab) (oratories) 10.10 6 CaoV3 cells were subcutaneously transplanted (6 individuals in each group). .

[0475] Intravenous administration of ADC1-C, reference ADC Ref-A, or ADC vehicle according to the present invention The treatment was performed when the tumor was approximately 150 mm in size. 3It was started when it reached [a certain level]. The animals received one injection (Q Treatment was performed with either 1d1) or 3 injections (once a week) (Q7d3). Tumor volume (Length × Width × Height × 0.52) is administered at least twice a week via electronic calipas after the initial injection. Measurements were taken for approximately 25 days. The results are shown in Figure 19 (animals treated with Q1d1) and Figure 20 (Q7 (Animals treated on day 3).

[0476] As can be seen in Figures 19 and 20, the ADC according to the present invention has remarkable effectiveness and Even after just one injection, complete tumor regression was observed.

[0477] 5. Complex formation between PNU-159682 derivative and monoclonal antibody 5.1. Synthesis, Purification, and Characterization of ADCs Two PNU-159682 derivatives, namely F562524 (Example J) and F 562646 (Example H) was subjected to the antibody 208F2 (A) under the conditions previously described in Example 4. It was coupled to b1) and c9G4(Ab2). 208F2(Ab1) and c9G4( Ab2) is as disclosed in Example 4. Briefly speaking, the antibody (1-5 mg / ml) This is mixed with a 10 mM borate buffer containing 150 mM NaCl and 2 mM EDTA. Partially reduce the solution with 6-20 equivalents of TCEP hydrochloride in a pH of 8.4 at 37°C for 2-4 hours. Next, the antibody concentration was measured using 150 mM NaCl, 2 mM EDTA, and 6% sucrose. Prepare the solution by adjusting the concentration to 1 mg / ml with 10 mM borate buffer at pH 8.4, and then add 10 mM DMS. From solution O, a drug linker complex in an excess of 5-20 molars relative to the antibody was added. The reaction proceeded as follows: The procedure was carried out at room temperature or at 37°C for 1 to 4 hours in the presence of 0% DMSO. Excess drugs were drug-induced. Add 2.5 moles of N-acetylcysteine ​​per liter and incubate at room temperature for 1 hour. The mixture was rapidly cooled by performing a suction. It was then left overnight at 4°C with 150 mM NaCl and 6% sucrose. After dialyzing with 25 mM His buffer pH 6.5 containing , ADC antibody was administered. Purified by chromatography or ultrafiltration. ADC concentration was measured using IgG as a standard. The purified ADC was determined using a BCA assay, and the 0.2 μm f After filtering and sterilizing, it was stored at 4°C.

[0478] To confirm drug complex formation and recrosslinking, and to determine the content of monomers and aggregates. To determine this, SDS-PAGE and SEC(T) were used as previously described in Example 4. ADCs were further analyzed using an SK G3000 SWXL column. Monomer content The figure was around 95% (Figure 23 and Table 13).

[0479] [Table 26]

[0480] 5.2. ADC Analysis using Native LC-MS UPLC Acqui connected to Synapt G2Si mass spectrometer (Waters) Native liquid chromatography-mass spectrometry using the ty H Class Bio system. ADC was analyzed using the following method. LC separation was performed using two polyhydroxyethyl A-C223 The procedure was performed using a Lamb (Poly-LC, 150×1mm, 300A, 5μm). The sample was eluted. Diluted to 0.2 mg / ml with buffer (150 mM ammonium acetate). 4 μg of s The sample was injected and eluted at a flow rate of 40 μL / min. The mass spectrometer was set to positive mode. The capillary was operated at a voltage of 2.9kV. The sample cone was set to 150V. The analysis was performed as follows: The scan was performed with a scan time of 1 second in the m / z range of 1000 to 8000. Figure 24 shows the decompression. The m / z spectrum before solution is shown. The DAR distribution was obtained using Mass Lynx software. Using Waters' MaxEnt™ algorithm, the MS spectrum is analyzed. Determined after convolution (Figure 25). The mean DAR value is given by the following formula (where j is the maximum drug). The calculation was performed using the number of physical loads.

number

[0481] [Table 27]

[0482] 5.3. In vitro stability ADC hz208F2-F562524 was administered to cynomolgus monkey serum at a concentration of 200 μg / ml. The samples were incubated at 37°C for 14 days. Samples were collected on days 0, 3, 7, and 14. The samples were then stored at -80°C until LC-MS analysis was performed to determine the average DAR.

[0483] Before LC-MS analysis, the sample was treated with Capture Select anti-human IgG-bio. With a tin complex (Life Technologies, 8 μg antibody / 200 μL beads) Coated streptavidin magnetic beads (M-280, Invitrogen) Immunopurification was performed using [a specific method]. The sample was placed in a chamber with anti-IgG coated beads. Incubate at warm temperature for 2 hours (100 μL sample / 200 μL beads), then 40 Acid elution was performed with μL of 0.4% trifluoroacetic acid. 4 μL of 3M Tris / HCl The pH was increased by adding a pH 8.8 solution. The immunopurified samples were then divided into two groups. Incubate with μL of IgGZero at 37°C for 30 minutes, then proceed as described above. LC-MS analysis was performed under non-denaturing conditions. The DAR distribution was obtained using Mass Lynx software. Using Waters' MaxEnt™ algorithm, the MS spectrum is analyzed. Determined after convolution, the mean DAR value is given by the following formula (where j is the maximum drug loading number). The calculation was performed using (ru).

number

[0484] [Table 28]

[0485] 5.4. In vitro cytotoxicity The cytotoxicity of ADCs was evaluated in MCF-7 cells and NCI-H2122 cells. The cells were seeded in complete growth medium in a 96-well plate (2500 cells / well). The next day, the experiment was conducted. A serially diluted solution of ADC was added to the corresponding wells and incubated at 37°C for 6 days. Cell survival rate is measured using the Cell Titer Glo kit (Promega) with ATP. It was determined by measurement. The luminescence was determined by Berthold Company's plate. The data was read using the Mithras application. The obtained data is expressed as a survival rate percentage. The results are shown in Figures 26 and 27. The survival rate of the untreated wells was set to 100%.

[0486] As expected, ADC hz208F2-F562524 and hz208F2- F562646 dramatically reduced cell viability. For NCI-H2122 cells: , 2 each for hz208F2-F562524 and hz208F2-F562646 .48.10 -11 M and 1.92.10 -12 M EC 50 The value is determined, MCF- For 7 cells, use hz208F2-F562524 and hz208F2-F5626 46: 5.86.10 -12 M and 9.45.10 -13 M EC 50 The value is It exhibits potent cytotoxic activity. In contrast, a countermeasure synthesized using unrelated antibodies ADCs exhibit moderate cytotoxicity in both MCF-7 and NCI-H2122 cells. It showed activity.

[0487] 5.5. In vivo antitumor activity 7-week-old female SCID mice (Charles River Laboratories) ) to 10.10 6 Caov3 cells were subcutaneously transplanted (6 individuals in each group). ADC hz2 08F2-F562524 (0.3 mg / kg), corresponding control ADC c9G4-F5 Treatment by 62524 (0.3 mg / kg) or intravenous administration of the vehicle (Q7d2). The tumor was approximately 150 mm 3 It was started when it reached [a certain value]. Tumor volume (length × width × height × 0. 52) was measured at least twice a week using an electronic caliper for approximately 25 days after the initial injection. The results are shown in Figure 19 (animals treated with Q1d1) and Figure 20 (animals treated with Q7d3). The results are shown in Figure 28. Complete recovery after two injections of ADC hz208F2-F562524. Tumor regression was observed, but no antitumor effect was seen with the control ADC or vehicle.

[0488] 5.6. Conclusion By using sulfomaleimide linker technology, the PNU-159682 derivative is used. The ADCs synthesized using this method are extremely homogeneous and stable in serum. Their efficacy varies. This was demonstrated in in vitro and in vivo models.

[0489] 6. Overall Conclusion Overall, the linker technology based on sulfomaleimide described herein is Phosphorus based on the usual maleimide used in marketed compounds such as Docetris Compared to Carr, it shows higher stability in the plasma of various species and in vitro models. It shows high efficacy. These characteristics are found in various cell lines, and more specifically, in cells with lower antigen expression. This resulted in a clear improvement in in vivo efficacy in the cell line (CAOV3).

[0490] Furthermore, the ADC according to the present invention is related to improved efficacy and safety margins in human therapy. As it is more stable in circulation, better tolerability is also expected.

Claims

1. Linker of equation (I) below: 【Chemistry 1】 [In the formula, X １ and X ２ These are H, halogen atoms, and (C) atoms, independently of each other. １ -C ６ ) represents an alkoxy, or optionally substituted aryloxy, however X １ and X ２ It cannot simultaneously represent H; L １ represents the group of the formula L １ ’-(CO-Z’) ｚ’ where L １ ’ is -(CH ２ ), -(CH ｎ CH ２ CH ２ O)-(CH ｍ -CH ２ -CH ２ -, arylene, heteroarylene, cycloalkanediyl, -(CH ２ )-arylene-, -(CH ｎ )-heteroarylene-, -(CH ２ )-cycloalkanediyl-, -arylene-(CH ｎ )-, -heteroarylene-(CH ２ )-, -cycloalkanediyl-(CH ｎ )-, -(CH ２ )-arylene-(CH<000A025>)-, -(CH ２ )-heteroarylene-(CH ｐ )-, -(CH ２ )-cycloalkanediyl-(CH ｐ )-, -(CH[[ID=A7]] ２ )-arylene-(CH ｎ )-, -(CH[[ID=5A]] ２ )-heteroarylene-(CH ｐ )-, -(CH ２ )-cycloalkanediyl-(CH ｎ )-, -(CH ２ CH ｐ O)-(CH<A000038>)-CH ｎ -CH ２ )-arylene-(CH ｐ )-, -(CH ２ CH ２ O)-(CH ｍ )-CH ２ -CH<A000046>)-arylene-(CH ２ )-, -(CH ｐ CH ２ CH ２ O)-(CH ｍ )-CH ２ -CH ２ )-heteroarylene-(CH ２ )-, -(CH ｐ CH ２ CH ２ O)-(CH ｍ )-CH ２ -CH ２ -Cycloalkanediyl- (CH ２ ) ｐ -, - (CH ２ ) ｎ -Arirene-CH ２ -CH ２ - (OCH ２ CH ２ ) ｍ -, - (CH ２ ) ｎ -Heterorialene-CH ２ -CH ２ - (OCH ２ CH ２ ) ｍ - or - (CH ２ ) ｎ -Cycloalkanediyl-CH ２ -CH ２ - (OCH ２ CH ２ ) ｍ - and; Each W independently represents an amino acid unit; Y is PAB-CO-(Z) ｚ -And here, PAB is, 【Chemistry 2】 Therefore, the oxygen in the PAB unit is CO-(Z) ｚ It is linked to; Z is -NR ４ -(CH ２ ) ｕ -NR<00,00089>-, -NR ４ -(CH ２ ) ｕ -NR ５ -CO-, -NR ４ -(CH ２ ) ｕ -NR ５ -CO-(CH ２ ) ｖ -, or -NR ４ -(CH ２ ) ｕ -NR ５ -CO-(CH ２ ) ｖ -CO-, and the NR ４ group is linked to the CO group of PAB-CO; Z' is -NR ４ - (CH ２ ) ｕ -NR ５ - or - NR ４ - (CH ２ ) ｕ -NR ５ -CO-(CH ２ ) ｖ - and the aforementioned NR ４ The group is linked to the CO group of CO-Z'; R ４ and R ５ H or (C) are independent of each other. １ -C ６ ) is an alkyl group; c is either 0 or 1; m is an integer between 1 and 15; n is an integer between 1 and 6; p is an integer between 1 and 6; q is 2; u is an integer between 1 and 6; v is an integer between 1 and 6; w is an integer between 0 and 5; y is either 0 or 1; z is either 0 or 1; z' is either 0 or 1; and X ３ This is when y = z = 1 and Z is -NR ４ - (CH ２ ) ｕ -NR ５ - If c=w=y=0, z'=1 and Z' is -NR ４ - (CH ２ ) ｕ -NR ５ It represents H when -, and X otherwise. ３ OH, NH ２ Alternatively, it represents a leaving group, where the leaving group is a halogen atom, formula -OSO ２ -R ＬＧ The sulfonates are N-succinimidyloxy, 4-nitrophenyloxy, pentafluorophenoxy or N-benzotriazoloxy, and R ＬＧ (C) may optionally be substituted with one or more halogen atoms. １ -C ６ ) Alkyl, aryl, aryl-(C １ -C ６ ) alkyl or (C １ -C ６ (Represents an alkyl-aryl group) or its salt.

2. The following formula (Ia): 【Transformation 3】 [In the formula, X １ , X ２ , X ３ , L １ , W, Y, c and w are as defined in claim 1, and, If w is 0, then y is 0; if w is an integer between 1 and 5, then y is 0 or 1. A linker or a salt thereof according to claim 1, having the characteristics of the linker or a salt thereof.

3. At least X １ or X ２ The linker according to claim 2, wherein represents a halogen atom.

4. At least X １ or X ２ The linker according to claim 3, wherein is represented by Br or Cl.

5. X １ and X ２ The linker according to claim 4, wherein one of the groups represents Br or Cl, and the other group represents H, Cl, or Br.

6. L １ 'ga- (CH ２ ) ｎ -, - (CH ２ CH ２ O) ｍ -CH ２ -CH ２ -, Arylene, -Cycloalkanediyl-, -(CH ２ ) ｎ -Arirene-, -Arirene- (CH ２ ) ｎ -, - (CH ２ ) ｎ -Cycloalkanediyl-, -Cycloalkanediyl-(CH ２ ) ｎ - 【Chemistry 4】 A linker according to claim 1, representing the above.

7. L １ 'ga- (CH ２ ) ｎ - or - (CH ２ CH ２ O) ｍ -CH ２ -CH ２ - The linker according to claim 6.

8. The linker according to claim 1, wherein each W is selected from alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, proline, lysine, lysine protected with acetyl or formyl, arginine, arginine protected with tosyl or a nitro group, histidine, ornithine, ornithine protected with acetyl or formyl, and citrulline.

9. w = 0 and (W) ｗ Is it a combination or w = 2 and (W) ｗ is Val-Cit or Val-Ala, The linker according to claim 1.

10. X １ and X ２ They are identical, and Cl, Br, (C １ -C ６ ) Alkoxy, and aryloxy (halogen, CN, NO ２ , and aryloxy groups which may be optionally substituted with one or more halogen atoms, or X １ and X ２ One of them is H, and the other is Cl, Br, (C １ -C ６ ) Alkoxy, and aryloxy (halogen, CN, NO ２ and optionally substituted with one or more groups selected from aryloxys which may optionally be substituted with one or more halogen atoms, The linker according to claim 1.

11. y = z = 1 and Z is -NR ４ - (CH ２ ) ｕ -NR ５ - If c=w=y=0, z'=1 and Z' is -NR ４ - (CH ２ ) ｕ -NR ５ - If X ３ H is H, and in other cases, X ３ The linker according to claim 1, wherein is OH, Cl, or N-succinimidyloxy.

12. The linker-drug complex of the following formula (II): 【Transformation 5】 [In the formula, X １ and X ２ These are H, halogen atoms, and (C) atoms, independently of each other. １ -C ６ ) represents an alkoxy, or optionally substituted aryloxy, however X １ and X ２ It cannot simultaneously represent H; L １ is, formula L １ '-(CO-Z)' ｚ’ This represents the base of, where L １ ' is, - (CH ２ ) ｎ -, - (CH ２ CH ２ O) ｍ -CH ２ -CH ２ -, Arylene, Heteroarylene, Cycloalkanediyl, - (CH ２ ) ｎ -Arirene-, -(CH ２ ) ｎ -Heterorialene-, -(CH ２ ) ｎ -Cycloalkanediyl-, -Arirene-(CH ２ ) ｐ -, -heterarylene-(CH ２ ) ｐ -,-cycloalkanediyl-(CH ２ ) ｐ -, - (CH ２ ) ｎ -Ariren- (CH ２ ) ｐ -, - (CH ２ ) ｎ -Heterorialene- (CH ２ ) ｐ -, - (CH ２ ) ｎ -Cycloalkanediyl- (CH ２ ) ｐ -, - (CH ２ CH ２ O) ｍ -CH ２ -CH ２ -Ariren- (CH ２ ) ｐ -, - (CH ２ CH ２ O) ｍ -CH ２ -CH ２ -Heterorialene- (CH ２ ) ｐ -, - (CH ２ CH ２ O) ｍ -CH ２ -CH ２ -Cycloalkanediyl- (CH ２ ) ｐ -, - (CH ２ ) ｎ -Arirene-CH ２ -CH ２ - (OCH ２ CH ２ ) ｍ -, - (CH ２ ) ｎ -Heterorialene-CH ２ -CH ２ - (OCH ２ CH ２ ) ｍ - or - (CH ２ ) ｎ -Cycloalkanediyl-CH ２ -CH ２ - (OCH ２ CH ２ ) ｍ - and; Each W independently represents an amino acid unit; Y is PAB-CO-(Z) ｚ -And here, PAB is, 【Transformation 6】 Therefore, the oxygen in the PAB unit is CO-(Z) ｚ It is linked to; Z is -NR ４ - (CH ２ ) ｕ -NR ５ -, -NR ４ - (CH ２ ) ｕ -NR ５ -CO-, -NR ４ - (CH ２ ) ｕ -NR ５ -CO-(CH ２ ) ｖ -, or -NR ４ - (CH ２ ) ｕ -NR ５ -CO-(CH ２ ) ｖ -CO- and the aforementioned NR ４ The group is linked to the CO group of PAB-CO; Z' is -NR ４ - (CH ２ ) ｕ -NR ５ - or - NR ４ - (CH ２ ) ｕ -NR ５ -CO-(CH ２ ) ｖ - and the aforementioned NR ４ The group is linked to the CO group of CO-Z'; R ４ and R ５ H or (C) are independent of each other. １ -C ６ ) is an alkyl group; Q represents the drug part; c is either 0 or 1; m is an integer between 1 and 15; n is an integer between 1 and 6; p is an integer between 1 and 6; q is 2; u is an integer between 1 and 6; v is an integer between 1 and 6; w is an integer between 0 and 5; y is either 0 or 1; z is 0 or 1; and z' is either 0 or 1. or its salt.

13. The following formula (IIa): 【Transformation 7】 [In the formula, X １ , X ２ , L １ , W, Y, Q, c and w are as defined in claim 12, and If w is 0, then y is 0; if w is an integer between 1 and 5, then y is 0 or 1. A linker drug complex according to claim 12, or a salt thereof, having the same.

14. L １ 'ga- (CH ２ ) ｎ - or - (CH ２ CH ２ O) ｍ -CH ２ -CH ２ - and; w = 0 and (W) ｗ Is it a combination or w = 2 and (W) ｗ is Val-Cit or Val-Ala; and X １ and X ２ They are identical, and Cl, Br, (C １ -C ６ ) Alkoxy, and aryloxy (halogen, CN, NO ２ , and aryloxy groups which may be optionally substituted with one or more halogen atoms, or X １ and X ２ One of them is H, and the other is Cl, Br, (C １ -C ６ ) Alkoxy, and aryloxy (halogen, CN, NO ２ and optionally substituted with one or more groups selected from aryloxys which may optionally be substituted with one or more halogen atoms, The linker-drug complex according to claim 12.

15. The linker-drug conjugate according to claim 12, wherein Q is a residue of auristatin, anthracycline, camptothecin, SN-38, tubulisin, calicheamicin, meitansinoid, duocalmycin, amanitin, pyrrolobenzodiazepine, or an immune checkpoint activator.

16. The linker-drug conjugate according to claim 15, wherein the immune checkpoint activator is a residue of an interferon gene (STING) agonist stimulant or a residue of an indoleamine 2,3-dioxygenase (IDO) inhibitor.

17. Q is, A residue of monomethyl auristatin F (MMAF), monomethyl auristatin E (MMAE), or monomethyl dorastatin-10, or the following formula (C): 【Transformation 8】 [In the formula, R １ is H or OH, R ２ is, (C １ -C ６ ) alkyl, COOH, COO-((C １ -C ６ )alkyl) or thiazolyl, R ３ is H or (C １ -C ６ ) is alkyl, X ４ is O or NR ９ And, R ９ is H or (C １ -C ６ ) is alkyl and t is an integer between 1 and 8. A residue of the derivative having the following Daunorubicin, doxorubicin, epirubicin, idarubicin, 2-pyrrolinodoxorubicin, pro-2-pyrrolinodoxorubicin, or residues of PNU-159682, or formula (A) or (B) below: 【Chemistry 9】 The residue of; Camptothecin or residues of SN-38; Residues of tubulosin A, tubulosin B, tubulosin C, or tubulosin D; Residues of esperamicin, calicheamicin γ1, or N-acetyldimethylhydrazide calicheamicin; Maytansine, DM1 or DM4 residue; Residues of duocalmycin A, duocalmycin B1, duocalmycin B2, duocalmycin C1, duocalmycin C2, duocalmycin D, duocalmycin SA, or CC-1065; Residues of α-amanitin, β-amanitin, γ-amanitin, or ε-amanitin; Residues of anthramycin or SGD-1882; Below formula (D): 【Chemistry 10】 [In the formula, X １１ and X ２１ They are independently O or S, X １２ and X ２２ These are independently OH, SH, O, or S. A １１ and A ２１ These are independently, formula: 【Chemistry 11】 It is the basis of, and in the formula, ・Z １ is OR １１ , NR １１ R １２ , O or NR １１ And here, R １１ and R １２ H and R are independent of each other. １３ or COR １３ And here, R １３ is, (C １ -C ６ ) alkyl, aryl or aryl (C １ -C ６ ) is alkyl, ・Z ２ H, NR ２１ R ２２ Or NR ２１ And here, R ２１ and R ２２ H and R are independent of each other. ２３ or COR ２３ And here, R ２３ is, (C １ -C ６ ) alkyl, aryl or aryl (C １ -C ６ ) is alkyl, ・Z ３ is N or CR ３３ And here, R ３３ is an H or halogen atom, and ・Z ４ is H or (C １ -C ６ ) is alkyl, A １２ and A ２２ These are independently H, OH, or F, and A ２ Is it H, or A? ２ and A ２２ They are interconnected, and here, A ２ CH ２ A ２２ is O, Here, ・X １２ If O or S, then X ２２ is not O, and not S, Z １ is not O, and NR １１ No, Z ２ NR ２１ Instead, the residue of the STING agonist is X １２ It is linked to the rest of the molecule by; ・X ２２ If O or S, then X １２ is not O, and not S, Z １ is not O, and NR １１ No, Z ２ NR ２１ Instead, the residue of the STING agonist is X ２２ It is linked to the rest of the molecule by; ・Z １ is O or NR １１ If X １２ It is not O, and not S, but X ２２ is not O, and not S, Z ２ NR ２１ Instead, the residue of the STING agonist is Z １ It is linked to the rest of the molecule by; ・Z ２ NR ２１ If X １２ It is not O, and not S, but X ２２ is not O, and not S, Z １ is not O, and NR １１ Instead, the residue of the STING agonist is Z ２ [Linked to the rest of the molecule by...] The residue of The linker-drug complex according to claim 12.

18. Q is Below formula (A): 【Chemistry 12】 Below formula (B): 【Chemistry 13】 Below formula (C): 【Chemistry 14】 [In the formula, R １ is H or OH, R ２ is, (C １ -C ６ ) alkyl, COOH, COO-((C １ -C ６ )alkyl) or thiazolyl, R ３ is H or (C １ -C ６ ) is alkyl, X ４ is O or NR ９ And, R ９ is H or (C １ -C ６ ) is alkyl and t is an integer from 1 to 8]; or Below formula (D): 【Chemistry 15】 [In the formula, X １１ and X ２１ They are independently O or S, X １２ and X ２２ These are independently OH, SH, O, or S. A １１ and A ２１ These are independently, formula: 【Chemistry 16】 It is the basis of, and in the formula, ・Z １ is OR １１ , NR １１ R １２ , O or NR １１ And here, R １１ and R １２ H and R are independent of each other. １３ or COR １３ And here, R １３ is, (C １ -C ６ ) alkyl, aryl or aryl (C １ -C ６ ) is alkyl, ・Z ２ H, NR ２１ R ２２ Or NR ２１ And here, R ２１ and R ２２ H and R are independent of each other. ２３ or COR ２３ And here, R ２３ is, (C １ -C ６ ) alkyl, aryl or aryl (C １ -C ６ ) is alkyl, ・Z ３ is N or CR ３３ And here, R ３３ is an H or halogen atom, and ・Z ４ is H or (C １ -C ６ ) is alkyl, A １２ and A ２２ These are independently H, OH, or F, and A ２ Is it H, or A? ２ and A ２２ They are interconnected, and here, A ２ CH ２ A ２２ is O, Here, ・X １２ If O or S, then X ２２ is not O, and not S, Z １ is not O, and NR １１ No, Z ２ NR ２１ Instead, the residue of the STING agonist is X １２ It is linked to the rest of the molecule by; ・X ２２ If O or S, then X １２ is not O, and not S, Z １ is not O, and NR １１ No, Z ２ NR ２１ Instead, the residue of the STING agonist is X ２２ It is linked to the rest of the molecule by; ・Z １ is O or NR １１ If X １２ It is not O, and not S, but X ２２ is not O, and not S, Z ２ NR ２１ Instead, the residue of the STING agonist is Z １ It is linked to the rest of the molecule by; ・Z ２ NR ２１ If X １２ It is not O, and not S, but X ２２ is not O, and not S, Z １ is not O, and NR １１ Instead, the residue of the STING agonist is Z ２ [Linked to the rest of the molecule by...] The linker drug complex according to claim 12, having the following characteristics.

19. Use of a linker according to any one of claims 1 to 11 or a drug linker complex according to any one of claims 12 to 18 for covalently linking a drug to a binding unit selected from a peptide, protein, antibody and its antigen-binding fragment.

20. Use of the linker or drug linker complex according to claim 19 for covalently linking a drug to a binding unit selected from an antibody and its antigen-binding fragment.

21. The following equation (III) or (IV): 【Chemistry 17】 [In the formula, The binding unit is a peptide, protein, antibody, or its antigen-binding fragment; L １ is, formula L １ '-(CO-Z)' ｚ’ This represents the base of, where L １ ' is, - (CH ２ ) ｎ -, - (CH ２ CH ２ O) ｍ -CH ２ -CH ２ -, Arylene, Heteroarylene, Cycloalkanediyl, - (CH ２ ) ｎ -Arirene-, -(CH ２ ) ｎ -Heterorialene-, -(CH ２ ) ｎ -Cycloalkanediyl-, -Arirene-(CH ２ ) ｐ -, -heterarylene-(CH ２ ) ｐ -,-cycloalkanediyl-(CH ２ ) ｐ -, - (CH ２ ) ｎ -Ariren- (CH ２ ) ｐ -, - (CH ２ ) ｎ -Heterorialene- (CH ２ ) ｐ -, - (CH ２ ) ｎ -Cycloalkanediyl- (CH ２ ) ｐ -, - (CH ２ CH ２ O) ｍ -CH ２ -CH ２ -Ariren- (CH ２ ) ｐ -, - (CH ２ CH ２ O) ｍ -CH ２ -CH ２ -Heterorialene- (CH ２ ) ｐ -, - (CH ２ CH ２ O) ｍ -CH ２ -CH ２ -Cycloalkanediyl- (CH ２ ) ｐ -, - (CH ２ ) ｎ -Arirene-CH ２ -CH ２ - (CH ２ CH ２ O) ｍ -, - (CH ２ ) ｎ -Heterorialene-CH ２ -CH ２ - (CH ２ CH ２ O) ｍ - or - (CH ２ ) ｎ -Cycloalkanediyl-CH ２ -CH ２ - (CH ２ CH ２ O) ｍ - and; Each W independently represents an amino acid unit; Y is PAB-CO-(Z) ｚ -And here, PAB is, [Chemistry 18] (The oxygen in the above PAB unit is CO-(Z) ｚ (and is connected to); Z is -NR ４ - (CH ２ ) ｕ -NR ５ -, -NR ４ - (CH ２ ) ｕ -NR ５ -CO-, -NR ４ - (CH ２ ) ｕ -NR ５ -CO-(CH ２ ) ｖ -, or -NR ４ - (CH ２ ) ｕ -NR ５ -CO-(CH ２ ) ｖ -CO- and the aforementioned NR ４ The group is linked to the CO group of PAB-CO; Z' is -NR ４ - (CH ２ ) ｕ -NR ５ - or - NR ４ - (CH ２ ) ｕ -NR ５ -CO-(CH ２ ) ｖ - and the aforementioned NR ４ The group is linked to the CO group of CO-Z'; R ４ and R ５ H or (C) are independent of each other. １ -C ６ ) is an alkyl group; Q represents the drug part; c is either 0 or 1; m is an integer between 1 and 15; n is an integer between 1 and 6; p is an integer between 1 and 6; s is an integer between 1 and 8; u is an integer between 1 and 6; v is an integer between 1 and 6; w is an integer between 0 and 5; y is either 0 or 1; z is 0 or 1; and z' is either 0 or 1. A drug conjugate unit, or a salt thereof.

22. L １ 'ga- (CH ２ ) ｎ - or - (CH ２ CH ２ O) ｍ -CH ２ -CH ２ - and; and w = 0 and (W) ｗ Is it a combination or w = 2 and (W) ｗ is Val-Cit or Val-Ala, The drug-binding unit complex according to claim 21.

23. The conjugation unit drug conjugate according to claim 21, wherein Q is a residue of auristatin, anthracycline, camptothecin, SN-38, tubulisin, calicheamicin, meitansinoid, duocalmycin, amanitin, pyrrolobenzodiazepine, or an immune checkpoint activator.

24. The binding unit is an antibody or its antigen-binding fragment, and The conjugated drug conjugate according to claim 21, wherein the antibody is an IGF-1R antibody or a HER2 antibody.

25. A pharmaceutical composition comprising a conjugated drug complex according to any one of claims 21 to 24 and at least one pharmaceutically acceptable excipient.

26. The pharmaceutical composition according to claim 25 for use in the treatment of cancer.

27. The pharmaceutical composition according to claim 26, wherein the cancer is prostate cancer, osteosarcoma, lung cancer, breast cancer, endometrial cancer, glioblastoma, colon cancer, gastric cancer, kidney cancer, pancreatic cancer, or head and neck cancer.

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