Glycoside-containing peptide linkers for antibody-drug conjugates

Abstract

To provide novel antibody-drug conjugate structures.SOLUTION: The present disclosure provides antibody-drug conjugate structures, which include a cleavable linker that links the antibody to the drug and has a first cleavable moiety and a second cleavable moiety that hinders cleavage of the first cleavable moiety. The disclosure also encompasses methods of production of such conjugates, as well as methods of using the same.SELECTED DRAWING: Figure 3

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation of U.S. Provisional Application No. 62 / 795,875, filed January 23, 2019. The benefit of which is claimed, and the disclosure of which provisional application is incorporated herein by reference. [Background technology]

[0002] Introduction The field of protein-small molecule therapeutic conjugates has made great advances and is expected to see even more in the future. It offers a number of clinically beneficial drugs with the promise of providing more. Quality conjugate therapeutics offer, for example, specificity, multiplicity of function, and fewer side effects. Offers several advantages due to the resulting relatively low off-target activity Chemical modifications of proteins can make them stronger, more stable, or more versatile. These advantages can be extended by Summary of the Invention [Problem to be solved by the invention]

[0003] The present disclosure provides a method for linking an antibody to a drug and a first cleavable moiety and a second cleavable moiety. antibody-drug combinations comprising a cleavable linker having a second cleavable moiety that prevents cleavage of the antibody-drug combination. The present disclosure also provides methods for producing such conjugates. In addition to the law, it also encompasses how to use it.

[0004] Embodiments of the present disclosure include an antibody, a drug, and a compound that links the antibody to the drug and has a first cleavable moiety. and a cleavable molecule having a second cleavable portion that prevents cleavage of the first cleavable portion. Conjugates containing a suitable linker are also included.

[0005] In some embodiments, the first cleavable moiety is an enzymatically cleavable moiety and the second The cleavable moiety of 2 is a chemically cleavable moiety.

[0006] In some embodiments, the first cleavable moiety is a chemically cleavable moiety and the second The two cleavable moieties are enzymatically cleavable moieties.

[0007] In some embodiments, the first cleavable moiety is a first enzymatically cleavable moiety; and the second cleavable moiety is a second enzymatically cleavable moiety. For example, the first enzyme The enzymatically cleavable moiety can comprise a first peptide and a second enzymatically cleavable The portion can include a second peptide. In some cases, the first enzymatically cleavable portion can include a second peptide. The moiety comprises a peptide and the second enzymatically cleavable moiety comprises a glycoside.

[0008] In some embodiments, the conjugate has formula (I): [ka]

[0009] wherein: Z is CR 4 or N, R 1 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, Substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl and substituted heterocyclyl; R 2 and R 3is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, a amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy , acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl aryl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, hetero aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; is R 2 and R 3 may be cyclically linked to form a 5- or 6-membered heterocyclyl. Ku, Each R 4 is hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, Amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy Acylamino, aminoacyl, alkylamido, substituted alkylamido, sulfo aryl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, hydroxyl Heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl , heterocyclyl, and substituted heterocyclyl; Each R 5 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl , alkynyl, and substituted alkynyl; Each R 6 is an alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkyn ... alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; k is an integer from 1 to 10, R 7 is hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, a amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy , acylamino, aminoacyl, alkylamido, substituted alkylamido, aryl , substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, Selected from substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl And, L 1 is the first linker, L 2 is a second linker, W 1 is a drug, and W 2 is an antibody, L 1 , R 6 or R 7 One of the fragments comprises a second cleavable portion.

[0010] In some embodiments, k is 2 and the conjugate has formula (Ia): [ka]

[0011] wherein: R 6’ or R 6’’one of which contains a second cleavable moiety, and R 6’ and R 6’’ The other of the groups is alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heteroaryl and substituted heterocyclyl.

[0012] In some embodiments, the second cleavable moiety is a glycoside-containing enzymatically cleavable It is a part.

[0013] In some embodiments, the conjugate has formula (Ib): [ka]

[0014] It is a conjugate of

[0015] In some embodiments, the conjugate has formula (Ic): [ka]

[0016] In some embodiments, k is 2 and the conjugate is the formula (Id): [ka]

[0017] is a conjugate of R 6’ and R 6’’ is an alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, hetero Aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heteroaryl independently selected from tetracyclyl, tetracyclyl, and substituted heterocyclyl; and R 7 comprises a second cleavable moiety.

[0018] In some embodiments, the second cleavable moiety is a glycoside-containing enzymatically cleavable It is a part.

[0019] In some embodiments, the conjugate has formula (Ie): [ka]

[0020] It is a conjugate of

[0021] In some embodiments, k is 2 and the conjugate has the formula (If): [ka]

[0022] is a conjugate of R 6’ and R 6’’ is an alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, hetero Aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heteroaryl independently selected from tetracyclyl, tetracyclyl, and substituted heterocyclyl; and R 8 comprises a second cleavable moiety.

[0023] In some embodiments, the conjugate has formula (Ig): [ka]

[0024] It is a conjugate of

[0025] In some embodiments, L 1 teeth, -(T 1 -V 1 ) a -(T 2 -V 2 ) b -(T 3 -V 3 ) c -(T 4 -V 4 ) d - wherein a, b, c, and d are each independently 0 or 1; T 1 , T 2 , T 3 and T 4 is a covalent bond, (C1~C 12 ) alkyl, substituted ( C1~C 12 ) alkyl, aryl, substituted aryl, heteroaryl, substituted Heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and Substituted heterocyclyl, (EDA) w , (PEG) n , (AA) p , -(CR 13 O H) m -, 4-amino-piperidine (4AP), acetal group, hydrazine, disulfide and esters, each independently selected from the group consisting of ethylenediamine moiety and ethylenediamine ester. where PEG is polyethylene glycol and AA is an amino acid residue, and each w is 1. each n is an integer from 1 to 30, and each p is an integer from 1 to 20; and each m is an integer from 1 to 12; V 1 , V 2 , V 3 and V 4 is a covalent bond, -CO-, -NR 15 -, -NR 15 (C H2) q -, -NR 15 (C6H4)-, -CONR 15 -, -NR 15 CO-, -C( O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR 15 -, -NR 15 Independently selected from the group consisting of SO2- and -P(O)OH- each q is an integer from 1 to 6; Each R 13 is hydrogen, alkyl, substituted alkyl, aryl, and substituted arylene. are selected independently from the rule, and Each R 15 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl , aryl, substituted aryl, heteroaryl, substituted heteroaryl, cyclo Alkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl are selected independently from the

[0026] In some embodiments, L 2 teeth, -(T 5 -V 5 ) e -(T 6 -V 6 ) f -(T 7 -V 7 ) g -(T 8 -V 8 ) h - wherein e, f, g, and h are each independently 0 or 1; T 5 , T 6 , T 7 and T 8 is a covalent bond, (C1~C 12 ) alkyl, substituted ( C1~C 12 ) alkyl, aryl, substituted aryl, heteroaryl, substituted Heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and Substituted heterocyclyl, (EDA) w , (PEG) n , (AA) p , -(CR 13 O H) m -, 4-amino-piperidine (4AP), acetal group, hydrazine, disulfide and esters, each independently selected from the group consisting of ethylenediamine moiety and ethylenediamine ester. where PEG is polyethylene glycol and AA is an amino acid residue, and each w is 1. each n is an integer from 1 to 30, and each p is an integer from 1 to 20; and each m is an integer from 1 to 12; V 5 , V 6 , V 7 and V 8 is a covalent bond, -CO-, -NR 15 -, -NR 15 (C H2) q -, -NR 15 (C6H4)-, -CONR 15 -, -NR 15 CO-, -C( O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR 15 -, -NR 15 Independently selected from the group consisting of SO2- and -P(O)OH- each q is an integer from 1 to 6; Each R 13 is hydrogen, alkyl, substituted alkyl, aryl, and substituted arylene. are selected independently from the rule, and Each R 15 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl , aryl, substituted aryl, heteroaryl, substituted heteroaryl, cyclo Alkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl are selected independently from the

[0027] In some embodiments, T 1 is (C1~C 12 ) alkyl and substituted (C1-C 12 ) alkyl Selected, T 2 , T 3 , and T 4 is an aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and and substituted heterocyclyl, (EDA) w , (PEG) n , (C1~C 12 )Archi substituted (C1-C 12 ) alkyl, (AA) p , -(CR 13 OH) m -, 4- Amino-piperidine (4AP), acetal group, hydrazine, and ester, respectively. are independently selected, and V 1 , V 2 , V 3 and V 4is a covalent bond, -CO-, -NR 15 -, -NR 15 (C H2) q -, -NR 15 (C6H4)-, -CONR 15 -, -NR 15 CO-, -C( O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR 15 -, -NR 15 Independently from the group consisting of SO2-, and -P(O)OH- Selected, (PEG) n teeth [ka]

[0028] and n is an integer from 1 to 30. EDA has the following structure: [ka]

[0029] wherein y is an integer from 1 to 6, and r is 0 or 1. and 4-amino-piperidine (4AP) [ka]

[0030] and Each R 12 and R 15 is hydrogen, alkyl, substituted alkyl, polyethyleneglycol any two adjacent aryl moieties independently selected from aryl and substituted aryl; R 12 the groups may be cyclically linked to form a piperazinyl ring, and R 13is hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. The rule is selected from the following:

[0031] In some embodiments, T 1 (C1~C 12 ) alkyl and V 1 is -CO-, T 2 is 4AP and V 2 is a covalent bond, T 3 (PEG) n and V 3 is -CO-, and d is 0.

[0032] In some embodiments, T 1 (C1~C 12 ) alkyl and V 1 is -CO-, T 2 is 4AP and V 2 is a covalent bond, T 3 (PEG) n and V 3 is -CONH-, and T 4 is aryl or substituted aryl, and V 4 is -CO-.

[0033] In some embodiments, T 5 is a covalent bond, and V 5 is -CO-, and f, g and h are 0.

[0034] In some embodiments, the drug is an auristatin, a maytansine, or a duocarmey. is selected from

[0035] An aspect of the present disclosure is a compound comprising a cleavable linker for linking an antibody to a drug. The cleavable linker is a first cleavable moiety and a second cleavable moiety. The compound includes a second cleavable moiety that interferes.

[0036] In some embodiments, the first cleavable moiety is an enzymatically cleavable moiety and the second The cleavable moiety of 2 is a chemically cleavable moiety.

[0037] In some embodiments, the first cleavable moiety is a chemically cleavable moiety and the second The two cleavable moieties are enzymatically cleavable moieties.

[0038] In some embodiments, the first cleavable moiety is a first enzymatically cleavable moiety; and the second cleavable moiety is a second enzymatically cleavable moiety. For example, the first enzyme The enzymatically cleavable moiety can be a first peptide and a second enzymatically cleavable The first enzymatically cleavable portion can be a second peptide. The moiety comprises a peptide and the second enzymatically cleavable moiety comprises a glycoside.

[0039] In some embodiments, the compound has formula (II): [ka]

[0040] is a compound of the formula Z is CR 4 or N, R 2 and R 3 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, a amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy , acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl aryl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, hetero aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; is R 2 and R 3 may be cyclically linked to form a 5- or 6-membered heterocyclyl. Ku, Each R 4 is hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, Amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy Acylamino, aminoacyl, alkylamido, substituted alkylamido, sulfo aryl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, hydroxyl Heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl , heterocyclyl, and substituted heterocyclyl; Each R 5 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl , alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl , substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclo and substituted heterocyclyl; Each R 6is an alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkyn ... alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; k is an integer from 1 to 10, R 7 is hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, a amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy , acylamino, aminoacyl, alkylamido, substituted alkylamido, aryl , substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, Selected from substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl And, L 1 is the first linker, L 2 is a second linker, and W 1 is a drug, L 1 , R 6 or R 7 One of the fragments comprises a second cleavable portion.

[0041] In some embodiments, k is 2 and the compound has formula (IIa): [ka]

[0042] is a compound of the formula R 6’ or R 6’’one of which contains a second cleavable moiety, and R 6’ and R 6’’ The other of the groups is alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heteroaryl and substituted heterocyclyl.

[0043] In some embodiments, the second cleavable moiety is a glycoside-containing enzymatically cleavable It is a part.

[0044] In some embodiments, the compound has the formula (IIb): [ka]

[0045] is a compound of

[0046] In some embodiments, the compound has the formula (IIc): [ka]

[0047] is a compound of

[0048] In some embodiments, k is 2 and the compound has formula (IId): [ka]

[0049] is a compound of the formula R 6’ and R 6’’ is an alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, hetero Aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heteroaryl independently selected from tetracyclyl, tetracyclyl, and substituted heterocyclyl; and R 7 comprises a second cleavable moiety.

[0050] In some embodiments, the second cleavable moiety is a glycoside-containing enzymatically cleavable It is a part.

[0051] In some embodiments, the compound has the formula (IIe): [ka]

[0052] is a compound of

[0053] In some embodiments, k is 2 and the compound has the formula (IIf): [ka]

[0054] is a compound of the formula R 6’ and R 6’’ is an alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, hetero Aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heteroaryl independently selected from tetracyclyl, tetracyclyl, and substituted heterocyclyl; and R 8 comprises a second cleavable moiety.

[0055] In some embodiments, the compound has the formula (IIg): [ka]

[0056] is a compound of

[0057] In some embodiments, L 1 teeth, -(T 1 -V 1 ) a -(T 2 -V 2 ) b -(T 3 -V 3 ) c -(T 4 -V 4 ) d - wherein a, b, c, and d are each independently 0 or 1; T 1 , T 2 , T 3 and T 4 is a covalent bond, (C1~C 12 ) alkyl, substituted ( C1~C 12 ) alkyl, aryl, substituted aryl, heteroaryl, substituted Heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and Substituted heterocyclyl, (EDA) w , (PEG) n , (AA) p , -(CR 13 O H) m -, 4-amino-piperidine (4AP), acetal group, hydrazine, disulfide and esters, each independently selected from the group consisting of ethylenediamine moiety and ethylenediamine ester. where PEG is polyethylene glycol and AA is an amino acid residue, and each w is 1. each n is an integer from 1 to 30, and each p is an integer from 1 to 20; and each m is an integer from 1 to 12; V1 , V 2 , V 3 and V 4 is a covalent bond, -CO-, -NR 15 -, -NR 15 (C H2) q -, -NR 15 (C6H4)-, -CONR 15 -, -NR 15 CO-, -C( O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR 15 -, -NR 15 Independently selected from the group consisting of SO2- and -P(O)OH- each q is an integer from 1 to 6; Each R 13 is hydrogen, alkyl, substituted alkyl, aryl, and substituted arylene. are selected independently from the rule, and Each R 15 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl , aryl, substituted aryl, heteroaryl, substituted heteroaryl, cyclo Alkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl are selected independently from the

[0058] In some embodiments, L 2 teeth, -(T 5 -V 5 ) e -(T 6 -V 6 ) f -(T 7 -V 7 ) g -(T 8 -V 8 ) h - wherein e, f, g, and h are each independently 0 or 1; T 5 , T 6 , T 7 and T 8 is a covalent bond, (C1~C 12 ) alkyl, substituted ( C1~C 12 ) alkyl, aryl, substituted aryl, heteroaryl, substituted Heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and Substituted heterocyclyl, (EDA) w , (PEG) n , (AA) p , -(CR 13 O H) m -, 4-amino-piperidine (4AP), acetal group, hydrazine, disulfide and esters, each independently selected from the group consisting of ethylenediamine moiety and ethylenediamine ester. where PEG is polyethylene glycol and AA is an amino acid residue, and each w is 1. each n is an integer from 1 to 30, and each p is an integer from 1 to 20; and each m is an integer from 1 to 12; V 5 , V 6 , V 7 and V 8 is a covalent bond, -CO-, -NR 15 -, -NR 15 (C H2) q -, -NR 15 (C6H4)-, -CONR 15 -, -NR 15 CO-, -C( O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR 15 -, -NR 15 Independently selected from the group consisting of SO2- and -P(O)OH- each q is an integer from 1 to 6; Each R13 is hydrogen, alkyl, substituted alkyl, aryl, and substituted arylene. are selected independently from the rule, and Each R 15 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl , aryl, substituted aryl, heteroaryl, substituted heteroaryl, cyclo Alkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl are selected independently from the

[0059] In some embodiments, T 1 is (C1~C 12 ) alkyl and substituted (C1-C 12 ) alkyl Selected, T 2 , T 3 , and T 4 is an aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and and substituted heterocyclyl, (EDA) w , (PEG) n , (C1~C 12 )Archi substituted (C1-C 12 ) alkyl, (AA) p , -(CR 13 OH) m -, 4- Amino-piperidine (4AP), acetal group, hydrazine, and ester, respectively. are independently selected, and V 1 , V 2 , V 3 and V 4 is a covalent bond, -CO-, -NR 15 -, -NR 15(C H2) q -, -NR 15 (C6H4)-, -CONR 15 -, -NR 15 CO-, -C( O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR 15 -, -NR 15 Independently from the group consisting of SO2-, and -P(O)OH- Selected, (PEG) n teeth [ka]

[0060] and n is an integer from 1 to 30. EDA has the following structure: [ka]

[0061] wherein y is an integer from 1 to 6, and r is 0 or 1. and 4-amino-piperidine (4AP) [ka]

[0062] and Each R 12 and R 15 is hydrogen, alkyl, substituted alkyl, polyethyleneglycol any two adjacent aryl moieties independently selected from aryl and substituted aryl; R 12 the groups may be cyclically linked to form a piperazinyl ring, and R 13 is hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. The rule is selected from the following:

[0063] In some embodiments, T 1 (C1~C 12 ) alkyl and V 1 is -CO-, T 2 is 4AP and V 2 is a covalent bond, T 3 (PEG) n and V 3 is -CO-, and d is 0.

[0064] In some embodiments, T 1 (C1~C 12 ) alkyl and V 1 is -CO-, T 2 is 4AP and V 2 is a covalent bond, T 3 (PEG) n and V 3 is -CONH-, and T 4 is aryl or substituted aryl, and V 4 is -CO-.

[0065] In some embodiments, T 5 is a covalent bond, and V 5 is -CO-, and f, g and h are 0.

[0066] In some embodiments, the drug is an auristatin, a maytansine, or a duocarmey. is selected from

[0067] An embodiment of the present disclosure is a pharmaceutical composition comprising a conjugate of the present disclosure and a pharmaceutically acceptable excipient: Pharmaceutical compositions include:

[0068] An aspect of the present disclosure is a method comprising administering to a subject an effective amount of a conjugate of the present disclosure. Includes:

[0069] An aspect of the present disclosure is a method of treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure. and administering a pharmaceutical composition comprising the conjugate of the present invention to a subject, wherein the administration The present invention includes a method for treating cancer by administering a compound to a subject, the compound being effective in treating cancer. [Brief explanation of the drawings]

[0070] [Figure 1A] FIG. 1A shows a graph of the serum stability over time of an antibody-drug conjugate containing an MMAE construct according to an embodiment of the present disclosure. [Figure 1B] FIG. 1B shows a graph of serum stability over time of an antibody-drug conjugate containing duocarmycin construct 66, according to an embodiment of the present disclosure. [Figure 2A] FIG. 2A shows a graph of the in vitro efficacy of antibody-drug conjugates containing MMAE constructs against the NCI-N87 cell line, according to an embodiment of the present disclosure. [Figure 2B] FIG. 2B shows a graph of the in vitro efficacy of antibody-drug conjugates containing duocarmycin construct 66 against the JeKo-1 cell line, according to an embodiment of the present disclosure. [Figure 3] FIG. 3 shows a graph of the in vivo efficacy of anti-CD79b 13 and 48 antibody-drug conjugates against JeKo-1 xenografts in mice according to an embodiment of the disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0071] definition The following terms have the following meanings unless otherwise indicated: Any Undefined Terms have their art-recognized meanings.

[0072] "Alkyl" refers to an alkyl group having 1 to 10 carbon atoms, for example, 1 to 6 carbon atoms, or 1 to 5 or monovalent saturated aliphatic hydrocarbyl groups having 1 to 4, or 1 to 3 carbon atoms The term refers to, by way of example, straight and branched chain hydrocarbyl groups, such as methyl n-propyl (CH3CH2CH2-), ethyl (CH3CH2CH2-), Isopropyl ((CH3)2CH-), n-butyl (CH3CH2CH2CH2-), isobutyl ((CH3)2CHCH2-), sec-butyl ((CH3)(CH3CH2) CH-), t-butyl ((CH3)3C-), n-pentyl (CH3CH2CH2CH2 CH2-), and neopentyl ((CH3)3CCH2-).

[0073] The term "substituted alkyl" refers to an alkyl group as defined herein. and one or more carbon atoms in the alkyl chain (excluding the C1 carbon atom) are heteroatoms. Child, e.g., -O-, -N-, -S-, -S(O) n -(n is 0 to 2), -NR- (R is hydrogen or alkyl), and may be substituted with alkoxy, substituted substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, aminoacyl aryl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxy aryl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thio heteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy hydroxyamino, alkoxyamino, nitro -SO-alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl -SO2-aryl, -SO2-aryl, -SO2-heteroaryl, and -NR a R b A group consisting of R has 1 to 5 substituents selected from ’ and R ’’ may be the same or different, and hydrogen, optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkane, It refers to a group selected from nyl, alkynyl, aryl, heteroaryl and heterocycle.

[0074] "Alkylene" is either linear or branched, preferably 1 to 6 alkylenes, more preferably Preferably, it has 1 to 3 carbon atoms, and is —O—, —NR 10 -, -NR 10 C(O)-, and -C(O)NR 10 -, etc., may be inserted. This term refers to a divalent aliphatic hydrocarbyl group. Examples include methylene (-CH2 -), ethylene (-CH2CH2-), n-propylene (-CH2CH2CH2-), i so-Propylene (-CH2CH(CH3)-), (-C(CH3)2CH2CH2-) , (-C(CH3)2CH2C(O)-), (-C(CH3)2CH2C(O)NH-) , and (-CH(CH3)CH2-), etc.

[0075] "Substituted alkylene" refers to the carbon described in the definition of "substituted" below. It refers to an alkylene group having 1 to 3 hydrogen atoms replaced by a substituent such as

[0076] The term "alkane" refers to alkyl and alkane groups as defined herein. Refers to the alkylene group.

[0077] "Alkylaminoalkyl", "alkylaminoalkenyl" and "alkylamino The term "alkynyl" refers to the group R'NHR''-, where R' is as defined herein. and R'' is an alkyl group as defined herein. The term "alkynylene" refers to a substituted or unsubstituted aryl group, which may be an aryl, alkenylene, or alkynylene group.

[0078] The term "alkaryl" or "aralkyl" refers to the groups -alkylene-aryl and -substituted alkylene-aryl, including alkylene, substituted alkylene and Aryl refers to as defined herein.

[0079] "Alkoxy" refers to the group -O-alkyl, where alkyl is defined herein. Examples of alkoxy include methoxy, ethoxy, n-propyl, and propyl. propoxy, isopropoxy, n-butoxy, t-butoxy, sec-butoxy, and n The term "alkoxy" also refers to the group alkenyl-O- , cycloalkyl-O—, cycloalkenyl-O—, and alkynyl-O—, Alkenyl, cycloalkyl, cycloalkenyl, and alkynyl are used herein to refer to Refers to something that is as defined.

[0080] The term "substituted alkoxy" refers to the group: substituted alkyl-O-, substituted Alkenyl-O-, substituted cycloalkyl-O-, substituted cycloalkenyl-O - and substituted alkynyl-O-, including substituted alkyl, substituted alkynyl, cycloalkyl, substituted cycloalkenyl and substituted alkenyl It refers to quinyl as defined herein.

[0081] The term "alkoxyamino" refers to the group -NH-alkoxy, where alkoxy is As defined in the specification.

[0082] The term "haloalkoxy" refers to the group alkyl-O-, where one or more haloalkoxy groups are present on the alkyl group. refers to a group in which multiple hydrogen atoms have been replaced by halo groups, such as trifluoromethoxy. Which groups are mentioned?

[0083] The term "haloalkyl" refers to a substituted alkyl group as defined above, but not to an alkyl group. It refers to a group in which one or more hydrogen atoms on the alkyl group have been replaced by a halo group. Examples of such groups are: Examples include, but are not limited to, fluoroalkyl groups such as trifluoromethyl, difluoromethyl, ethyl, and trifluoroethyl.

[0084] The term "alkylalkoxy" refers to the group -alkylene-O-alkyl, alkylene- O-substituted alkyl, substituted alkylene-O-alkyl, and substituted alkene-O-alkyl alkylene-O-substituted alkyl, including alkyl, substituted alkyl, alkylene and substituted alkylene as defined herein.

[0085] The term "alkylthioalkoxy" refers to the group -alkylene-S-alkyl, alkylene substituted alkylene-S-alkyl and substituted alkylene-S-alkyl alkylene-S-substituted alkyl, including alkyl, substituted alkyl, alkylene The term "alkylene" refers to alkylene and substituted alkylene as defined herein.

[0086] "Alkenyl" has 2 to 6 carbon atoms, preferably 2 to 4 carbon atoms, and Linear or branched chains having at least one, preferably 1 to 2, double bond unsaturation sites This term refers to hydrocarbyl groups, examples of which include bi-vinyl, allyl, and buta-3 The cis and trans isomers or mixtures of these isomers are not included in this is included in the term.

[0087] The term "substituted alkenyl" includes alkoxy, substituted alkoxy, cycloalkoxy, Alkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl , acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, acyl aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, o oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thiohetero aryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thio Alkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, Tetracyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, - SO-Alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl alkyl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl and -SO2-heteroaryl, or 1 to 5 substituents selected from "Alkenyl" refers to an alkenyl group, as defined herein, having an alkyl group.

[0088] "Alkynyl" has 2 to 6 carbon atoms, preferably 2 to 3 carbon atoms, and Linear or branched chain with at least one, preferably one to two, triple bond unsaturation sites It refers to a linear monovalent hydrocarbyl group. Examples of such alkynyl groups are acetylenyl and (-C≡CH), and propargyl (-CHC≡CH).

[0089] The term "substituted alkynyl" refers to alkoxy, substituted alkynyl, cycloalkoxy, Alkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl , acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, acyl aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, o oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thiohetero aryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thio Alkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, Tetracyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, - SO-Alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl alkyl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl, and and -SO2-heteroaryl, or 1 to 5 substituents selected from It refers to an alkynyl group, as defined herein, having the following structure:

[0090] "Alkynyloxy" refers to the group -O-alkynyl, where alkynyl is as defined herein Alkynyloxy refers to those as defined above. Examples of alkynyloxy include ethynyloxy. oxy, and propynyloxy.

[0091] "Acyl" refers to the groups HC(O)-, alkyl-C(O)-, substituted alkyl-C( O)-, alkenyl-C(O)-, substituted alkenyl-C(O)-, alkynyl-C(O)- (O)-, substituted alkynyl-C(O)-, cycloalkyl-C(O)-, substituted cycloalkyl-C(O)-, cycloalkenyl-C(O)-, substituted cycloalkenyl aryl-C(O)-, aryl-C(O)-, substituted aryl-C(O)-, hetero Aryl-C(O)-, substituted heteroaryl-C(O)-, heterocyclyl-C( O)- and substituted heterocyclyl-C(O)-, including alkyl, substituted Alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, silyl Cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkene aryl, substituted aryl, heteroaryl, substituted heteroaryl, heteroaryl, heteroaryl Examples of heterocyclic rings include substituted heterocyclic rings and substituted heterocyclic rings as defined herein. For example, acyl includes the "acetyl" group CH3C(O)-.

[0092] "Acylamino" refers to the group -NR 20 C(O) alkyl, -NR 20 C(O) substituted Alkyl, NR 20 C(O)cycloalkyl, -NR 20 C(O)-substituted cycloalkenyl Kill, -NR 20 C(O)cycloalkenyl, -NR 20 C(O)-substituted cycloalkenyl Kenyl, -NR20 C(O)alkenyl, -NR 20 C(O) substituted alkenyl, - NR 20 C(O)alkynyl, -NR 20 C(O)-substituted alkynyl, -NR 20 C (O)aryl, -NR 20 C(O)-substituted aryl, -NR 20 C(O) Heteroa Reel, -NR 20 C(O)-substituted heteroaryl, -NR 20 C(O) heterocycle, and Yobi-NR 20 C(O)-substituted heterocycles, R 20 is hydrogen or alkyl and alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl , substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl aryl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, Substituted heteroaryl, heterocycle, and substituted heterocycle are defined herein. It refers to something that is as it appears to be.

[0093] The term "aminocarbonyl" or "aminoacyl" refers to the group -C(O)NR 21 R 22 and R 21 and R 22 is hydrogen, alkyl, substituted alkyl, alkenyl aryl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted Aryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted Cycloalkenyl, heteroaryl, substituted heteroaryl, heterocycle, and substituted and R 21 and R 22But they are bound to together with the attached nitrogen to form a heterocyclic or substituted heterocyclic group, or alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, Substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted The heteroaryl, heterocycle, and substituted heterocycle are as defined herein. It refers to something that is.

[0094] "Aminocarbonylamino" is the group -NR 21 C(O)NR 22 R 23 and R 2 1 , R 22 , and R 23 is independently selected from hydrogen, alkyl, aryl or cycloalkyl or two R groups joined to form a heterocyclyl group; Point.

[0095] The term "alkoxycarbonylamino" refers to the group -NRC(O)OR, where each R are independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, or is heterocyclyl, alkyl, substituted alkyl, aryl, heteroaryl, and heterocyclyl is as defined herein.

[0096] The term "acyloxy" refers to the group alkyl-C(O)O-, substituted alkyl-C(O)O-, (O)O-, cycloalkyl-C(O)O-, substituted cycloalkyl-C(O)O- , aryl-C(O)O-, heteroaryl-C(O)O-, and heterocyclyl-C(O)O- (O)O- is alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl alkyl, aryl, heteroaryl, and heterocyclyl are defined herein. It refers to something that is as it is.

[0097] "Aminosulfonyl" refers to the group -SO2NR 21 R 22 and R 21 and R 22 but , hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl alkyl, substituted alkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, heteroaryl, Independently selected from the group consisting of substituted heteroaryl, heterocycle, and substituted heterocycle. , R 21 and R 22 are linked together with the nitrogen attached thereto to form a heterocyclic or substituted The alkyl, substituted alkyl, alkenyl, substituted substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl Cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted Aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted hetero The ring refers to a ring as defined herein.

[0098] "Sulfonylamino" refers to the group -NR 21 SO2R 22 and R 21 and R 22 but , hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl alkyl, substituted alkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, heteroaryl, Independently selected from the group consisting of substituted heteroaryl, heterocycle, and substituted heterocycle. Selected and R 21 and R 22 are linked together with the atoms attached thereto to form a heterocyclic or may form a substituted heterocyclic group, and may be alkyl, substituted alkyl, alkyl Alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl , substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, and Substituted heterocycle refers to a heterocycle as defined herein.

[0099] "Aryl" or "Ar" refers to a single ring (e.g., as in a phenyl group) or or ring systems with multiple condensed rings (examples of such aromatic ring systems are naphthyl, anthryl, monovalent aromatic carbocyclic group of 6 to 18 carbon atoms having a ring structure, including aryl and indanyl wherein the fused ring may or may not be aromatic, provided that The point refers to an atom in an aromatic ring. This term is used, for example, in phenyl and naphthyl. Unless otherwise constrained by the definition of the aryl substituent, such aryl groups include: Acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, Alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted alkoxy oxy, substituted alkenyl, substituted alkynyl, substituted cycloalkyl, substituted substituted cycloalkenyl, amino, substituted amino, aminoacyl, acylamino, acylamino, alkyl, aryl, aryloxy, azido, carboxyl, carboxylalkyl, cyano, halogen, nitro, heteroaryl, heteroaryloxy, heterocyclyl, Heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, -SO-alkoxy, alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -S O2-Alkyl, -SO2-substituted alkyl, -SO2-aryl, -SO2-hetero 1 to 5 substituents selected from haloaryl and trihalomethyl, or 1 to 3 substituents It may be substituted with a substituent.

[0100] "Aryloxy" refers to the group -O-aryl, where aryl is, for example, phenoxy. hydroxy, and naphthoxy, and the like, and also substituted as defined herein. The term "aryl" refers to any group as defined herein, including optionally aryl groups.

[0101] "Amino" refers to the group -NH2.

[0102] The term "substituted amino" refers to the group -NRR, where each R is hydrogen, alkyl, Substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkynyl, substituted and independently from the group consisting of alkynyl, aryl, heteroaryl, and heterocyclyl. where at least one R is not hydrogen.

[0103] The term "azido" refers to the group -N3.

[0104] "Carboxyl", "carboxy" or "carboxylate" refers to -CO2H or its It refers to the salt of

[0105] "Carboxyl ester" or "carboxy ester" or "carboxy alkyl" The term "carboxyl" or "carboxylalkyl" refers to the group -C(O)O-alkyl, -C (O)O-substituted alkyl, -C(O)O-alkenyl, -C(O)O-substituted Alkenyl, -C(O)O-alkynyl, -C(O)O-substituted alkynyl, -C( -O)O-aryl, -C(O)O-substituted aryl, -C(O)O-cycloalkyl , -C(O)O-substituted cycloalkyl, -C(O)O-cycloalkenyl, -C( -O)O-substituted cycloalkenyl, -C(O)O-heteroaryl, -C(O)O- Substituted heteroaryl, -C(O)O-heterocycle, and -C(O)O-substituted heteroaryl A heterocyclic ring, which may be selected from alkyl, substituted alkyl, alkenyl, substituted alkenyl, a Alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cyclo Alkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl Heteroaryl, substituted heteroaryl, heterocycle, and substituted heterocycle are used herein. Refers to something that is as defined.

[0106] "(Carboxylic ester)oxy" or "carbonate" refers to the group -OC(O)O -alkyl, -OC(O)O-substituted alkyl, -OC(O)O-alkenyl, -OC(O)O-substituted alkenyl, -OC(O)O-alkynyl, -OC( O)O-substituted alkynyl, -OC(O)O-aryl, -OC(O)O-substituted -OC(O)O-substituted aryl, -OC(O)O-cycloalkyl, -OC(O)O-substituted cycloalkyl -OC(O)O-cycloalkyl, -OC(O)O-cycloalkenyl, -OC(O)O-substituted cycloalkenyl chloroalkenyl, -OC(O)O-heteroaryl, -OC(O)O-substituted heteroaryl Heteroaryl, -OC(O)O-heterocycle, and -OC(O)O-substituted heterocycle and wherein the alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkyl alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl , substituted heteroaryl, heterocycle, and substituted heterocycle are defined herein. It refers to something that is as it is.

[0107] "Cyano" or "nitrile" refers to the group --CN.

[0108] "Cycloalkyl" refers to single or multiple cyclic rings, including fused, bridged, and spiro ring systems. Examples of suitable cycloalkyl groups include: Examples include adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, and and cyclooctyl. Examples of such cycloalkyl groups include: Single ring structures, such as cyclopropyl, cyclobutyl, cyclopentyl, and cyclobutyl octyl, etc., or multiple ring structures, such as adamantanyl, etc.

[0109] The term "substituted cycloalkyl" refers to alkyl, substituted alkyl, alkoxy, alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azide, Cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxyl alkoxy alkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thio aryl, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, hetero Aryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxya amino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, - SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-substituted 1 to 5 selected from alkyl, -SO2-aryl and -SO2-heteroaryl or a cycloalkyl group having 1 to 3 substituents.

[0110] "Cycloalkenyl" refers to a group having single or multiple rings and at least one double bond. and preferably a non-aromatic cyclic alkyl group having 3 to 10 carbon atoms and having 1 to 2 double bonds. It refers to the aryl group.

[0111] The term "substituted cycloalkenyl" includes alkoxy, substituted alkoxy, Cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl phenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl aryl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxy aryl, keto, thioketo, carboxyl, carboxyl alkyl, thioaryloxy, thio Heteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy , heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro , -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-hetero Aryl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl and and -SO2-heteroaryl, or 1 to 5 substituents selected from It refers to a cycloalkenyl group having a group.

[0112] "Cycloalkynyl" refers to a group having single or multiple rings and at least one triple bond. It refers to a non-aromatic cycloalkyl group of 5 to 10 carbon atoms having the formula:

[0113] "Cycloalkoxy" refers to -O-cycloalkyl.

[0114] "Cycloalkenyloxy" refers to -O-cycloalkenyl.

[0115] "Halo" or "halogen" refers to fluoro, chloro, bromo, and iodo.

[0116] "Hydroxy" or "hydroxyl" refers to the group --OH.

[0117] "Heteroaryl" refers to a heteroaryl having 1 to 15 carbon atoms in the ring, e.g., 1 to 10 carbon atoms. and 1 to 10 heteroatoms selected from the group consisting of oxygen, nitrogen, and sulfur Such heteroaryl groups are those having a single ring (e.g., pyridinyl, imino). azolyl or furyl) or multiple condensed rings in the ring system (e.g., indolizinyl, quinolyl, In the case of groups such as linyl, benzofuran, benzimidazolyl or benzothienyl The ring system may have a ring structure in which at least one ring is aromatic. To satisfy this requirement, any heteroatom in such a heteroaryl ring may be replaced by H or a substituent, For example, it may be attached to an alkyl group or other substituent as described herein. In certain embodiments, the heteroaryl group may or may not be (one or more) The nitrogen and / or sulfur ring atoms may be oxidized to N-oxide (N → O), sulfur The term may also provide a pyridinyl, phenyl, or sulfonyl moiety, for example, pyridinyl. Heteroaryl substituents include aryl, pyrrolyl, indolyl, thiophenyl, and furanyl. Unless otherwise constrained by the definition, such heteroaryl groups may also be acyloxy, hydroxyloxy, hydroxypropyl ... Hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cyclo Alkyl, cycloalkenyl, substituted alkyl, substituted alkoxy, substituted Alkenyl, substituted alkynyl, substituted cycloalkyl, substituted cycloalkenyl aryl, amino, substituted amino, aminoacyl, acylamino, alkaryl, aryl aryl, aryloxy, azido, carboxyl, carboxyl alkyl, cyano, halogen , nitro, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy , aminoacyloxy, oxyacylamino, thioalkoxy, substituted thioalkoxy oxy, thioaryloxy, thioheteroaryloxy, -SO-alkyl, -SO-substituted substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl and -SO2-heteroaryl; and trihalomethyl, or substituted with 1 to 5 substituents, or 1 to 3 substituents selected from It may be done.

[0118] The term "heteroaralkyl" refers to the group -alkylene-heteroaryl, Aminoalkyl and heteroaryl are as defined herein. Examples include pyridylmethyl, pyridylethyl, and indolylmethyl.

[0119] "Heteroaryloxy" refers to -O-heteroaryl.

[0120] "Heterocycle", "heterocyclic", "Heterocycloalkyl" and "heterocyclyl" refer to groups consisting of a single ring, or fused bridged and heterocyclic rings. and spirocyclic systems, having multiple condensed rings and containing 1 to 10 heteroatoms, It refers to a saturated or unsaturated group having 20 ring atoms. These ring atoms are nitrogen, sulfur, or oxygen, and if in a fused ring system, one or more of the rings may be selected from cycloalkyl, a It can be aryl, or heteroaryl, provided that the point of attachment is through a non-aromatic ring. In certain embodiments, the nitrogen(s) and / or Sulfur atoms can be oxidized to give N-oxide, -S(O)-, or -SO2- moieties. To satisfy valence requirements, any heteroatom in such a heterocyclic ring may be , one or more H, or one or more substituents, such as those described herein. It may or may not be attached to such an alkyl group or other substituent.

[0121] Examples of heterocycles and heteroaryls include azetidine, pyrrole, imidazole, pyridine, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole Indoles, dihydroindoles, indazoles, purines, quinolizines, isoquinolizines quinoxaline, quinazoline, cinnoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline , pteridine, carbazole, carboline, phenanthridine, acridine, phenanthridine Lorin, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazole Imidazolidine, imidazoline, piperidine, piperazine, indoline, phthalidine amide, 1,2,3,4-tetrahydroisoquinoline, 4,5,6,7-tetrahydrobenzyl benzo[b]thiophene, thiazole, thiazolidine, thiophene, benzo[b]thiophene , morpholinyl, thiomorpholinyl (also called thiamorpholinyl), 1,1-dioxo Examples include isothiomorpholinyl, piperidinyl, pyrrolidine, and tetrahydrofuranyl. Examples include, but are not limited to:

[0122] Unless otherwise constrained by the definition of a heterocyclic substituent, such heterocyclic groups may be alkoxylated or alkoxylated. alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azide, Cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxyl alkoxy alkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thio aryl, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, hetero Aryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxya amino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, - SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-substituted Selected from alkyl, -SO2-aryl, -SO2-heteroaryl, and fused heterocycles The alkyl group may be substituted with 1 to 5, or 1 to 3, substituents.

[0123] "Heterocyclyloxy" refers to the group -O-heterocyclyl.

[0124] The term "heterocyclylthio" refers to the group heterocycle-S-.

[0125] The term "heterocyclene" refers to a heterocycle formed from a heterocycle, as defined herein. refers to a diradical group that is

[0126] The term "hydroxyamino" refers to the group --NHOH.

[0127] "Nitro" refers to the group -NO2.

[0128] "Oxo" refers to the atom (=O).

[0129] "Sulfonyl" refers to the group -SO2-alkyl, -SO2-substituted alkyl, -SO2 -Alkenyl, -SO2-substituted alkenyl, -SO2-cycloalkyl, -SO2 -substituted cycloalkyl, -SO2-cycloalkenyl, -SO2-substituted cyclo -Alkenyl, -SO2-aryl, -SO2-substituted aryl, -SO2-hetero Aryl, -SO2-substituted heteroaryl, -SO2-heterocycle, and -SO2- Substituted heterocycles, including alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkenyl Alkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl , heteroaryl, substituted heteroaryl, heterocycle, and substituted heterocycle are included in the present invention. As used herein, sulfonyl refers to a group as defined in the specification. Examples of sulfonyl include methyl. -SO2-, phenyl-SO2-, and 4-methylphenyl-SO2-.

[0130] "Sulfonyloxy" refers to the group -OSO2-alkyl, -OSO2-substituted alkyl , -OSO2-alkenyl, -OSO2-substituted alkenyl, -OSO2-cycloalkenyl alkyl, -OSO2-substituted cycloalkyl, -OSO2-cycloalkenyl, -O SO2-substituted cycloalkenyl, -OSO2-aryl, -OSO2-substituted Aryl, -OSO2-heteroaryl, -OSO2-substituted heteroaryl, -O SO2-heterocycles and -OSO2-substituted heterocycles, which may be alkyl, substituted Alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, silyl Cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkene aryl, substituted aryl, heteroaryl, substituted heteroaryl, heteroaryl, heteroaryl heterocycle, and substituted heterocycle are as defined herein.

[0131] "Sulfate" or "sulfuric acid ester" is a group -O-SO2-OH, -O-SO2- O-Alkyl, -O-SO2-O-substituted alkyl, -O-SO2-O-alkenyl , -O-SO2-O-substituted alkenyl, -O-SO2-O-cycloalkyl, -O -SO2-O-substituted cycloalkyl, -O-SO2-O-cycloalkenyl, -O -SO2-O-substituted cycloalkenyl, -O-SO2-O-aryl, -O-SO 2-O-substituted aryl, -O-SO2-O-heteroaryl, -O-SO2-O- Substituted heteroaryl, -O-SO2-O-heterocycle, and -O-SO2-O-substituted Heterocycles containing alkyl, substituted alkyl, alkenyl, substituted alkene, Nyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl , cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heptyl Heteroaryl, substituted heteroaryl, heterocycle, and substituted heterocycle are described herein. refers to something that is as defined in

[0132] The term "aminocarbonyloxy" refers to the group -OC(O)NRR, where each R is: independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, or polyaryl; alkyl, substituted alkyl, aryl, heteroaryl and heterocyclic rings It refers to what is defined herein.

[0133] "Thiol" refers to the group --SH.

[0134] The term "thioxo" or "thioketo" refers to the atom (=S).

[0135] The term "alkylthio" or "thioalkoxy" refers to the group -S-alkyl , where alkyl is as defined herein. In some cases, sulfur may be oxidized to -S(O)-. Sulfoxides may have one or more stereoisomers. It may exist as a sex form.

[0136] The term "substituted thioalkoxy" refers to the group -S-substituted alkyl.

[0137] The term "thioaryloxy" refers to the group aryl-S-, where the aryl group is The aryl groups defined herein include those optionally substituted aryl groups as defined herein. It refers to something that is as it is defined.

[0138] The term "thioheteroaryloxy" refers to the group heteroaryl-S- The aryl group also includes optionally substituted aryl groups as defined herein. It refers to what is defined herein.

[0139] The term "thioheterocyclooxy" refers to the group heterocyclyl-S- Cyclyl groups also include optionally substituted heterocyclyl groups as defined herein. "A" refers to a substance or a mixture of substances or compounds, including those defined herein.

[0140] Further to the disclosure herein, the term "substituted" refers to a group or group that is designated. When used to modify a radical, one or more of the specified groups or radicals The hydrogen atoms are each independently the same or different as defined below. It can also mean being substituted with a substituent.

[0141] In addition to the groups disclosed for each individual term herein, the specified group or radical may also be used. One or more hydrogens on saturated carbon atoms in the carbon (any two hydrogens on a single carbon = O, =NR 70 , =N-OR 70 , which can be replaced by =N2 or =S) The substituents are -R unless otherwise specified. 60 , halo, =O, -OR 70 , -SR 70 , -NR 80 R 80 , trihalomethyl, -CN, -OCN, -SCN, -NO, -NO2 , =N2, -N3, -SO2R 70 , -SO2O - M + , -SO2OR 70 , -OSO2 R 70 , -OSO2O - M + , -OSO2OR 70 , -P(O)(O - )2(M + )2, -P(O)(OR 70 )O - M + , -P(O)(OR 70 )2, -C(O)R 70 , -C (S)R 70 , -C(NR 70 )R 70 , -C(O)O - M + , -C(O)OR 70 ,- C(S)OR 70 , -C(O)NR 80 R 80 , -C(NR 70 )NR 80 R 80 , -O C(O)R 70 , -OC(S)R 70 , -OC(O)O - M + , -OC(O)OR 70 , -OC(S)OR 70 , -NR 70 C(O)R 70 , -NR 70C(S)R 70 , -NR 70 CO2 - M + , -NR 70 CO2R 70 , -NR 70 C(S)OR 70 , -NR 70 C(O)NR 80 R 80 , -NR 70 C(NR 70 )R 70 and -NR 70 C(NR 7 0 )NR 80 R 80 and R 60 is an optionally substituted alkyl, cycloalkyl , heteroalkyl, heterocycloalkylalkyl, cycloalkylalkyl, aryl , arylalkyl, heteroaryl, and heteroarylalkyl and each R 70 are independently hydrogen or R 60 and each R 80 is independently R 70 and or alternatively, two R 80’ together with the nitrogen atom to which they are attached, and S, and containing 1 to 4 additional heteroatoms, which may be the same or different, selected from the group consisting of may also form a 5-, 6-, or 7-membered heterocycloalkyl, and among the heteroatoms, N may have —H or C1-C3 alkyl substitution, and each M + is a net single positive charge Each M is a counterion with a charge. + are independently, for example, alkali ions, e.g., K + , Na + , Li + ammonium ions, e.g., + N(R 60) 4; or alkali Earth ions, e.g., [Ca 2+ ] 0.5 , [Mg 2+ ] 0.5 , or [Ba 2+ ]0 .5 (The subscript 0.5 indicates the counterion of such a divalent alkaline earth ion.) is one of the ionized forms of the compounds of the present invention and other typical counterions, e.g., chloride or the two ionized compounds disclosed herein can be such Can serve as a counterion to divalent alkaline earth ions or doubly ionized The compounds of the present invention can serve as counterions to such divalent alkaline earth ions. A specific example is -NR 80 R 80 is -NH2, -NH-alkyl N-pyrrolidinyl, N-piperazinyl, 4N-methyl-piperazin-1-yl and It is meant to include N-morpholinyl.

[0142] In addition to the disclosure herein, "substituted" alkenes, alkynes, aryls and Substituents for hydrogen on unsaturated carbon atoms in heteroaryl groups are -R, unless otherwise specified. 60 , halo, -O - M + , -OR 70 , -SR 70 , -S - -M + , -NR 80 R 80 , Trihalomethyl, -CF3, -CN, -OCN, -SCN, -NO, -NO2, -N3, -SO2R 70 , -SO3 - M + , -SO3R 70 , -OSO2R 70 , -OSO3- M + 、-OSO3R 70 、-PO3 -2 (M + )2、-P(O)(OR 70 )O - M + 、- P(O)(OR 70 )2、-C(O)R 70 、-C(S)R 70 、-C(NR 70 )R 7 0 、-CO2 - M + 、-CO2R 70 、-C(S)OR 70 、-C(O)NR 80 R 80 、-C(NR 70 )NR 80 R 80 、-OC(O)R 70 、-OC(S)R 70 、-OC O2 - M + 、-OCO2R 70 、-OC(S)OR 70 、-NR 70 C(O)R 70 、- NR 70 C(S)R 70 、-NR 70 CO2 - M + 、-NR 70 CO2R 70 、-NR 7 0 C(S)OR 70 、-NR 70 C(O)NR 80 R 80 、-NR 70 C(NR 70 )R 70 および-NR 70 C(NR 70 )NR 80 R 80 であり、R 60, R 70 , R 80 oh Yobi M + is as previously defined, except that in the case of a substituted alkene or alkyne, In this case, the substituent is -O - M + , -OR 70 , -SR 70 , and -S - M + Neither of the above stomach.

[0143] In addition to the groups disclosed for each individual term herein, "substituted" heteroaryls are also Substituents for hydrogen on nitrogen atoms in alkyl and cycloheteroalkyl groups are, unless otherwise specified, Otherwise, -R 60 , -O - M + , -OR 70 , -SR 70 , -S - M + , -NR 80 R 8 0 , trihalomethyl, -CF3, -CN, -NO, -NO2, -S(O)2R 70 , -S (O)2O - M + , -S(O)2OR 70 , -OS(O)2R 70 , -OS(O)2O - M + , -OS(O)2OR 70 , -P(O)(O - )2(M + )2, -P(O)(OR 7 0 )O - M + , -P(O)(OR 70 )(OR 70 ), -C(O)R 70 , -C(S)R 70 , -C(NR 70 )R 70, -C(O)OR 70 , -C(S)OR 70 , -C(O) NR 80 R 80 , -C(NR 70 )NR 80 R 80 , -OC(O)R 70 , -OC(S) R 70 , -OC(O)OR 70 , -OC(S)OR 70 , -NR 70 C(O)R 70 ,- NR 70 C(S)R 70 , -NR 70 C(O)OR 70 , -NR 70 C(S)OR 70 , -NR 70 C(O)NR 80 R 80 , -NR 70 C(NR 70 )R 70 and -NR 70 C(NR 70 )NR 80 R 80 and R 60 , R 70 , R 80 and M + was previously defined It is as stated.

[0144] Further to the disclosure herein, in certain embodiments, the substituted group is 1, 2, 3 or 4 substituents, 1, 2 or 3 substituents, 1 or 2 substituents, Or has one substituent.

[0145] All of the substituted groups defined above may themselves be substituted with further substituents. The polymers arrived at by defining the substituents (e.g., substituted aryl groups) and substituted aryl groups which are themselves substituted as substituents. aryl groups having substituted aryl groups) are not intended to be included herein. In such cases, the maximum number of such substitutions is three. The sequential substitution of substituted aryl groups specifically contemplated in the specification is Ru-(substituted aryl)--Limited to substituted aryl.

[0146] Unless otherwise indicated, substituent names not expressly defined herein are intended to be used interchangeably with the official names. This is reached by naming the terminal portion of the functional group, followed by the adjacent functional group to the point of attachment. For example, the substituent "arylalkyloxycarbonyl" is a group (aryl)-(alkyl )-OC(O)-.

[0147] It is understood that for any group disclosed herein that contains one or more substituents, Such groups are not intended to be limiting unless any substitution or substitution is sterically impractical and / or synthetically feasible. It is understood that the subject compounds do not contain these chemical or substitution patterns. This invention includes all stereochemical isomers arising from the substitution of the compounds.

[0148] The term "pharmaceutically acceptable salt" refers to a compound that is acceptable for administration to a patient, e.g., a mammal. Salts (salts with counterions that have acceptable mammalian safety for a given dosing regime) ) Such salts are salts of pharmaceutically acceptable inorganic or organic bases and pharmaceutically acceptable salts of It can be derived from an acceptable inorganic or organic acid. The term "salt" refers to a pharmaceutically acceptable salt of the compound, including various organic and non-organic salts well known in the art. and inorganic counterions, such as, by way of example only, sodium, potassium, calcium, magnesium, ammonium, ammonium, and tetraalkylammonium, and the molecules are basic If it contains a functional group, it may be a salt of an organic or inorganic acid, for example, hydrochloride, hydrobromide, formate. , tartrate, besylate, mesylate, acetate, maleate, and oxalate salts. be.

[0149] The term "salt thereof" refers to a salt of an acid in which the proton of the acid is substituted with a cation, e.g., a metal cation or an organic cation. means the compound formed when the cation is replaced by an ion, etc., where applicable The salts are pharmaceutically acceptable salts, but this does not include intermediate compounds that are not intended for administration to patients. For example, salts of the present compounds include those in which the compound is an inorganic or organic acid. protonated by an inorganic or organic acid to form a cation and as the anionic component of the salt. Examples include those having a conjugate base of the formula:

[0150] A "solvate" is a complex formed by the combination of solvent molecules with molecules or ions of a solute. The solvent can be an organic compound, an inorganic compound, or a mixture of both. Some examples of solvents are methanol, N,N-dimethylformamide, tetrahydrofuran, Examples of solvents include, but are not limited to, ethanol, dimethyl sulfoxide, and water. When the solvate formed is a hydrate.

[0151] "Stereoisomers" and "stereoisomers" are two molecules that have the same atom connectivity but different positions in space. It refers to compounds with the same atomic configuration. Stereoisomers include cis-trans isomers, E and These include Z and Z isomers, enantiomers, and diastereomers.

[0152] "Tautomers" are compounds that differ only in the electronic bonding of their atoms and / or the position of their protons. Alternative forms of the molecule, such as enol-keto and imine-enamine tautomers, or heteroaryl groups containing the -N=C(H)-NH- ring atom configuration, e.g., pyrazole, Tautomerism of benzoxazole, imidazole, benzimidazole, triazole, and tetrazole Those skilled in the art will recognize that other tautomeric ring atom arrangements are possible.

[0153] The term "or a salt or solvate or stereoisomer thereof" includes salts, solvates and and all permutations of stereoisomers, including pharmaceutically acceptable stereoisomers of the subject compounds. It will be appreciated that the term "solvates" is intended to include solvates of salts thereof.

[0154] "Pharmaceutically effective amount" and "therapeutically effective amount" refer to a dose or amount of a compound or compound that is effective for the treatment of a specified disorder or disease or its To treat one or more symptoms and / or prevent the occurrence of a disease or disorder refers to an amount of a compound sufficient to produce a pharmaceutically or therapeutically effective The amount may be, inter alia, sufficient to cause tumor shrinkage or reduce the rate of tumor growth. Contains sufficient amount.

[0155] "Patient" refers to human and non-human subjects, particularly mammalian subjects.

[0156] The terms "treat" or "treatment" as used herein refer to a patient, For example, to treat a disease or medical condition in a mammal (especially a human) or to Treatment means (a) preventing the occurrence of a disease or medical condition, e.g., (b) prophylactic treatment of a subject; (b) ameliorating a disease or medical condition, e.g., (c) eliminating or causing the regression of a disease or medical condition; For example, slowing or stopping the development of a disease or medical condition in a patient. (d) inhibiting a disease or medical condition in a patient; This includes relieving the symptoms of a medical condition.

[0157] The terms "polypeptide," "peptide," and "protein" are used herein to refer to The terms "amino acid" and "amino acid" are used interchangeably to refer to polymeric forms of amino acids of any length unless otherwise specified. Without this, "polypeptides," "peptides," and "proteins" are genetically coded Coded and non-coded amino acids, chemically or biochemically modified or derivatized These polypeptides may include polypeptides with modified amino acids, as well as polypeptides with modified peptide backbones. The term includes fusion proteins, which include fusion proteins with heterologous amino acid sequences. Proteins, fusions with heterologous and homologous leader sequences, at least one N-terminal methionine residues (e.g., to facilitate production in recombinant host cells), and Examples of suitable proteins include, but are not limited to, immunologically tagged proteins.

[0158] "Native amino acid sequence" or "parent amino acid sequence" are used interchangeably herein. and the amino acid sequence of the polypeptide before modification to include the modified amino acid residue. Point to the column.

[0159] Terms such as "amino acid analog" and "unnatural amino acid" may be used interchangeably. and naturally occurring proteins (e.g., Ala or A, Cys or C, As p or D, Glu or E, Phe or F, Gly or G, His or H, Il e or I, Lys or K, Leu or L, Met or M, Asn or N, Pr o or P, Gln or Q, Arg or R, Ser or S, Thr or T, Va I or V, Trp or W, Tyr or Y) Includes amino acid-like compounds that are similar in structure and / or overall shape to amino acids. Acid analogs also include natural amino acids with modified side chains or backbones. Amino acid analogs also include amino acids that have the same stereochemistry as the naturally occurring D-form. In addition to analogs, L-amino acid analogs are also included. The log shares the backbone structure and / or side chain structure of one or more natural amino acids, and ( The difference(s) is / are one or more modified groups in the molecule. indicates the substitution of a related atom (e.g., S) by an atom (e.g., N), a group (e.g., methyl, or Addition of an atom (e.g., xyl) or atom (e.g., Cl or Br), deletion of a group, or substitution of a covalent bond (such as replacing a double bond with a single bond), or combinations thereof. For example, amino acid analogs include, but are not limited to, α-hydroxy acids and α-amino acids. It may include, etc.

[0160] The terms "amino acid side chain" or "side chain of an amino acid" and the like refer to naturally occurring amino acids, non-naturally occurring amino acids, and the like. Substituents attached to the α-carbon of amino acid residues, including amino acids and amino acid analogs The amino acid side chains may also be referred to as modified amino acids as described herein. The amino acid may contain an amino acid side chain as described in the context of the acid and / or conjugate. Cut.

[0161] Terms such as "carbohydrate" refer to the monomer units of monosaccharides, disaccharides, oligosaccharides, and polysaccharides and The term sugar is sometimes used to refer to smaller carbohydrates and / or polymers. The term "carbohydrate derivatives" is sometimes used to refer to carbohydrates, e.g., monosaccharides, disaccharides. The term "carbohydrate" refers to a carbohydrate in which one or more functional groups of the carbohydrate of interest are substituted (in any convenient manner). replaced by a substituent), modified (changed to another group using any convenient chemistry) replaced) or absent (e.g., eliminated or replaced by H) A wide variety of carbohydrates and carbohydrate derivatives are available, including the subject compounds and It may be adapted for use in a conjugate.

[0162] The term "glycoside" or "glycosyl" refers to a group of amino acids attached to a moiety via a glycosidic bond. For example, glycosidic moieties are those described herein. The glycosidic bond can be a cleavable linker such that it can be easily cleaved. In this case, examples include, but are not limited to, O-glycosidic bonds (O-glycosides), N-glycosidic bonds, etc. glycosidic bond (glycosylamine), S-glycosidic bond (thioglycoside), or C-glycosidic bond glycosides to other moieties through glycosidic bonds (C-glycosidic or C-glycosyl) In some cases, glycosides can be linked to the glycosides by chemically mediated methods, e.g. from the moiety to which they are attached by hydrolysis or enzymatically mediated hydrolysis. It can be disconnected.

[0163] The term "antibody" is used in the broadest sense and includes monoclonal antibodies (full-length monoclonal antibodies). clonal antibodies), polyclonal antibodies, and multispecific antibodies (e.g., bispecific antibodies). heterologous antibodies), humanized antibodies, single chain antibodies, chimeric antibodies, and antibody fragments (e.g., Fab fragments ) and the like. Antibodies can bind to target antigens. (Janeway, C., Tr avers, P., Walport, M., Shlomchik (2001) Immun. o Biology,5th Ed.,Garland Publishing,New The target antigen is a complementarity determinant formed by one or more variable regions of an antibody. It has one or more binding sites, also called epitopes, recognized by the CDRs. It is possible.

[0164] The term "native antibody" refers to the heavy and light chains of an antibody produced by the immune system of a multicellular organism. The spleen, lymph nodes, bone marrow, and serum are tissues that produce natural antibodies. For example, antibody-producing cells isolated from a first animal immunized with an antigen The antibodies produced by are natural antibodies.

[0165] The term "humanized antibody" or "humanized immunoglobulin" refers to the corresponding modification of a human antibody. amino acids at positions other than those specified (e.g., in framework regions, constant regions, or CDRs) refers to a non-human (e.g., mouse or rabbit) antibody containing one or more amino acids of Generally, humanized antibodies are more effective in a human host than non-humanized versions of the same antibody. Antibodies can be produced using a variety of techniques known in the art. Such techniques include, for example, CDR grafting (EP grafting). 239,400; PCT International Publication No. 91 / 09967; U.S. Patent No. 5,225,539 No. 5,530,101, and No. 5,585,089) , veneering or resurfacing g)(EP 592,106;EP 519,596;Padlan, Molecule r Immunology 28(4 / 5):489-498(1991);Studn icka et al., Protein Engineering 7(6):805 -814(1994);Roguska.et al.,PNAS 91:969-97 3 (1994)), and chain shuffling (U.S. Pat. No. 5,565,332 In certain embodiments, framework substitutions are important for antigen binding. Analysis of the interactions between CDR and framework residues to identify key framework residues modeling and to identify unusual framework residues at specific positions (See, e.g., U.S. Pat. No. 5,585,089; Rie (See Schmann et al., Nature 332:323 (1988)). Additional methods for humanizing antibodies contemplated for use in the invention are described in U.S. Pat. Specification No. 50,078; Specification No. 5,502,167; Specification No. 5,705,154 Specification; Specification No. 5,770,403; Specification No. 5,698,417; Specification No. 5,6 Specification No. 93,493; Specification No. 5,558,864; Specification No. 4,935,496 and PCT International Publication No. 98 / 453. 31 and 98 / 45332. In certain embodiments, the subject rabbit The antibody is described in U.S. Patent Application Publication No. 20040086979 and U.S. Patent Application Publication No. It may also be humanized according to the methods set forth in the specification of US Pat. No. 20050033031. Thus, the above antibodies may be humanized using methods well known in the art.

[0166] The term "chimeric antibody" refers to an antibody whose light and heavy chain genes may belong to different species. It refers to antibodies constructed from mutant and constant region genes, typically by genetic engineering. For example, the variable segments of the genes from a mouse monoclonal antibody are substituted with human constant segments. Examples of therapeutic chimeric antibodies include those derived from mouse guanosine monophosphate (Guanidine diphosphate) and mouse guanosine monophosphate (Guanidine diphosphate) and those derived from ... variable or antigen-binding domains from a mouse antibody and constant or effector domains from a human antibody It is a hybrid protein composed of domains, but it does not contain domains from other mammalian species. A .sigma. may be used.

[0167] Immunoglobulin polypeptides Immunoglobulin light or heavy chain variable regions are "complementarity determining The framework region is flanked by three hypervariable regions, also called "CDRs" or "CDRs." The framework regions and CDRs are defined ( ''Sequences of Proteins of Immunology Interest,''E. Kabat et al.,USDepartmen (See t of Health and Human Services, 1991). The antibody framework, which is the combination of the framework regions of the constituent light and heavy chains. The nucleotide sequence of the CDRs is the region that serves to position and align the CDRs. This is the main cause of bonding.

[0168] A "parent Ig polypeptide" is an Ig polypeptide that contains an aldehyde-tagged constant region as described herein. The parent polypeptide is a polypeptide comprising an amino acid sequence lacking the native sequence. It may contain constant regions or may contain pre-existing amino acid sequence modifications (additions, deletions and / or substitutions) The constant region may comprise a constant region having a sequence such as

[0169] As used herein, the term "isolated" means that the compound of interest is It is meant to describe the compound in an environment other than that in which it naturally occurs. "Substantially enriched in the compound of interest and / or partially enriched in the compound of interest" means It is meant to include compounds that are present in a partially or substantially purified sample.

[0170] As used herein, the term "substantially purified" refers to a substance that is substantially purified from its native state. Removed from the environment and free of at least 60% of the other components with which it is naturally found At least 75% free, At least 80% free, At least All are 85% free, at least 90% free, at least 95% free free of, at least 98% free of, or more than 98% free of, It refers to a mixture.

[0171] The term "physiological conditions" refers to conditions that are compatible with living cells, e.g., temperatures that are compatible with living cells. It is meant to encompass primarily aqueous conditions such as temperature, pH, salinity, etc.

[0172] The "reactive partner" allows a molecule to react specifically with another reactive partner to produce a reaction product. The term "reactive partner" refers to a molecule or molecular moiety that produces a hydroxyl group. Exemplary reactive partners include sulfonyl groups, cysteine ​​or serine and formylglycine generating enzyme (FGE) of the catalysis motif These react to replace the cysteine ​​or serine in the motif with a hormone. Formation of a reaction product containing a converted aldehyde tag containing fluoroglycine (FGly) Other exemplary reactive partners include the fGly residue of the converted aldehyde tag. Aldehydes (e.g., reactive aldehyde groups), and aldehyde-reactive groups and compounds of interest The polypeptide contains a modified fGly moiety and is reacted to form a modified fGly residue. Reaction of the modified aldehyde-tagged polypeptide with a moiety of interest conjugated to a The "aldehyde-reactive reactive partner" forms a product.

[0173] "N-terminus" refers to the terminal amino acid residue of a polypeptide having a free amine group, The amine groups in the terminal amino acid residues usually form part of the covalent backbone of the polypeptide. It refers to.

[0174] The "C-terminus" refers to the terminal amino acid residue of a polypeptide having a free carboxyl group. The carboxyl groups in the non-C-terminal amino acid residues are linked through the covalent backbone of the polypeptide. Refers to something that is normally formed.

[0175] "Internal site" when used in reference to a polypeptide or the amino acid sequence of a polypeptide " refers to a region of a polypeptide that is neither N-terminal nor C-terminal.

[0176] Before further describing the present invention, it is to be understood that this invention is not limited to particular embodiments described, and that It should be understood that the terms used herein may vary. The following description is for purposes of describing particular embodiments only and is not intended to be limiting. It should also be understood that the scope of the present invention is limited only by the appended claims.

[0177] Where a range of values ​​is provided, the difference between the upper and lower limits of the range shall be such that the context clearly dictates otherwise. Each intervening value, to the tenth of the unit of the lower limit if not specified, and its stated range It is understood that any other stated or intervening values ​​within are encompassed by the present invention. The upper and lower limits of these smaller ranges shall not exceed any specifically excluded limit in the stated range. Each of the compounds may be independently included within this smaller range, subject to the degree of inertness, and also be included within the scope of the present invention. Where the stated range includes one or both of the limits, Ranges excluding either or both degrees are also included in the present invention.

[0178] Certain features of the invention are, for clarity, described in the context of separate embodiments. It is also recognized that these may be provided in combination in a single embodiment. Various features of the invention that are, for convenience, described in the context of a single embodiment, may also be provided separately. or in any suitable subcombination. The combinations are intended to be illustrative and not restrictive, and each and every combination may be used to produce, for example, a stable compound (i.e., A compound that can be isolated, characterized, and tested for biological activity To the extent that the invention encompasses subject matter that is a combination of two or more combinations, such combinations shall be treated as if they were individually and expressly disclosed. As such, they are specifically encompassed by the present invention and disclosed herein. and its elements (e.g., the compounds listed in the embodiments describing such variables). All subcombinations of the elements of the foundation of learning are also included in each and every such subcombination. All such combinations are specifically embraced by the present invention as if each were individually expressly disclosed herein. Disclosed in the specification.

[0179] Unless otherwise defined, all technical and scientific terms used herein refer to the invention It has the same meaning as commonly understood by a person skilled in the art to which it pertains. Any methods and materials similar or equivalent to those described may be used in the practice or testing of the present invention. Although any method and material can be used, the preferred methods and materials are now described. All publications mentioned in this document describe the methods and / or materials in the context in which the publications are referenced. No. 6,023,799, filed Dec. 1, 2003, incorporated herein by reference for purposes of disclosure and description.

[0180] As used in this specification and the appended claims, the singular forms "a," "an," and "an" are used interchangeably. It should be noted that "the" and "the" include plural referents unless the context clearly dictates otherwise. The claims must be drafted to exclude any optional element. It is further noted that this statement may be inconsistent with the recitation of claim elements. The use of exclusive academic terms such as "solely" and "only" in use, or serving as a prior basis for a "negative" qualification is intended.

[0181] Certain features of the invention are, for clarity, described in the context of separate embodiments. It is also recognized that these may be provided in combination in a single embodiment. Various features of the invention that are, for convenience, described in the context of a single embodiment, may also be provided separately. Or may be provided in any suitable subcombination.

[0182] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein shall be construed as to its authenticity by virtue of prior invention. Nothing contained herein should be construed as an admission that the Company does not have any prior rights to such publication. The published publication dates may be different from the actual publication dates, which may be independently confirmed. It may be necessary to

[0183] Detailed Description The present disclosure provides antibody-drug conjugates comprising a cleavable linker linking the antibody to the drug. The cleavable linker comprises a first cleavable moiety and a first cleavable The present disclosure also provides methods for treating such conjugates comprising the step of: In addition to the method of making the duplicate, the present invention also includes a method of using the duplicate.

[0184] Antibody-drug conjugates The present disclosure provides conjugates, e.g., antibody-drug conjugates (ADCs). By "conjugate" is meant a compound in which a first moiety (e.g., an antibody) is attached to a second moiety (e.g., a drug or active agent). For example, an antibody-drug conjugate The gates are designed to detect drugs (e.g., maytansine, auristatin or duocarmycin active agent moieties). By this, it is meant that the moiety is bound to another moiety or structure under standard conditions. In certain embodiments, the first and second moieties comprise one or more functional groups and a covalent bond. For example, one or more functional groups and a covalent bond may be bonded to one another through The cleavable linker may include a cleavable linker as described in the literature.

[0185] In certain embodiments, the conjugate comprises a polypeptide (e.g., a polypeptide conjugated to a second moiety) In certain embodiments, the polypeptide conjugate comprises a polypeptide The moiety conjugated to the peptide can be any of a variety of moieties of interest, including but not limited to: , a detectable label, a drug, a water-soluble polymer, or fixation of the polypeptide to a membrane or surface In certain embodiments, the conjugate can be a moiety for stabilization. and a drug conjugate wherein the polypeptide is an antibody, such that the antibody-drug conjugate For example, a conjugate can be a polypeptide conjugated to a drug or active agent moiety. For example, the drug or active agent may be maytansine. "Maytansine," "maytansine moiety," "maytansine active agent moiety," etc. "Maytansinoids" and "maytansinoids" refer to maytansine and its analogs and derivatives, including and the pharmaceutically active maytansine moiety and / or portions thereof. Conjugated maytansine may be any of a variety of maytansinoid moieties, including but not limited to: For example, maytansine and its analogs and derivatives as described herein. (e.g., deacylmaytansine) In other cases, the drug or active agent may be an auristatin or an analog thereof. or derivatives, or pharmaceutically active auristatin moieties and / or portions thereof. Auristatins conjugated to polypeptides can be prepared using a variety of auristatin moieties. any of the following, including but not limited to, auristatins and those described herein: In other cases, the drug or active ingredient may be an analog or derivative thereof, such as The agent may be a duocarmycin, or an analog or derivative thereof, or a pharmaceutically active and / or portions thereof. The duocarmycins conjugated to the amide may be any of a variety of duocarmycin moieties, including but not limited to: Non-limiting examples include duocarmycins and their analogs as described herein. It can be a log and a derivative.

[0186] In certain embodiments, the conjugate comprises an antibody and a drug linked by a linker. In some cases, the linker is a cleavable linker. A cleavable linker is a linker that contains one or more cleavable moieties, The cleavable moiety dissociates under certain conditions to separate the cleavable linker into two or more It contains one or more bonds that can be separated into more separable parts, for example: The cleavable moiety dissociates or decomposes under certain conditions to form two or more cleavable linkers. It may contain one or more covalent bonds that can be separated into one or more moieties. Therefore, the cleavable linker can be cleaved under appropriate conditions to allow the drug to act on the desired target. In the antibody-drug conjugate, the drug is separated or released from the antibody at the application site. can be included in

[0187] In some cases, the cleavable linker comprises two cleavable moieties, e.g., a first cleavable moiety The cleavage of both cleavable moieties results in the release of the drug. as needed to separate or release the drug from the antibody at the desired target site of action. For example, cleavage of the cleavable linker can be achieved by First, cut one of the two cleavable parts, then cut the other of the two cleavable parts. In certain embodiments, the cleavable linker can be one cleavable moiety and a second cleavable moiety that prevents cleavage of the first cleavable moiety By "prevents cleavage" it is meant that the presence of a second cleavable moiety that is not cleaved prevents the first 1, thereby reducing the likelihood of or substantially inhibiting cleavage of the cleavable portion of It is meant to substantially reduce the amount of available linker or prevent its cleavage. For example, the presence of an uncleaved second cleavable moiety may be indicative of the enzymatic cleavage of the first cleavable moiety. and / or chemical cleavage can be prevented by the presence of a second cleavable moiety. Preventing cleavage of the first cleavable moiety then substantially reduces the amount of drug from the antibody. For example, premature release of a drug from an antibody can be prevented by using an antibody-drug conjugate. gating is substantially reduced or prevented until the gating is at or near the desired target site of action of the drug. It can be done.

[0188] In some cases, the second cleavable moiety interferes with cleavage of the first cleavable moiety. Cleavage of the cleavable linker first cleaves the second cleavable moiety, then the first cleavage This can be achieved by cleaving the second cleavable portion. , reducing or eliminating interference with cleavage of the first cleavable moiety to allow the first cleavable moiety to be cleaved. The cutting of the first cleavable portion can be performed by As described above, the cleavable linker dissociates or separates into two or more parts to form an antibody. - This can result in the release of the drug from the drug conjugate. In some cases, cleavage of a first cleavable portion results in the formation of an uncleaved second cleavable portion. substantially does not occur in the presence of about 10% or that cleavage below that occurs in the presence of a second cleavable moiety that is not cleaved, e.g. For example, about 9% or less, or about 8% or less of the first cleavable portion, or about 7% or less, or about 6% or less, or about 5% or less less than, or about 4% or less, or about 3% or less, or about 2 % or less, or about 1% or less, or about 0.5% or less less than, or about 0.1% or less of the cuts are present in the uncut second cuttable portion. It is meant to occur in the presence of

[0189] In other words, the second cleavable moiety protects the first cleavable moiety from cleavage. For example, the presence of an uncleaved second cleavable moiety can inhibit the activity of the first cleavable moiety. The antibody-drug conjugate is protected from cleavage to target the drug to the desired site of action or Substantially reducing or preventing premature release of drug from the antibody until Therefore, cutting the second cleavable portion exposes the first cleavable portion. (e.g., by deprotecting the first cleavable moiety) so that the first cleavable moiety is cleaved. This results in cleavage of the cleavable linker, which then , as described above, to detach or release the drug from the antibody at the drug's desired target site of action. In certain instances, cleavage of the second cleavable moiety results in subsequent cleavage of the first cleavable moiety. cleavage of the second cleavable moiety, but cleavage of the second cleavable moiety does not itself does not result in cleavage of the linker (i.e., to cleave the cleavable linker) (Cleavage of the first cleavable portion is still required to achieve this.)

[0190] Each cleavable moiety contained in the cleavable linker is a chemically cleavable moiety. For example, the first cleavable moiety may be a chemically or enzymatically cleavable moiety. and the second cleavable moiety is chemically cleavable. In other cases, the first cleavable moiety can be an enzymatically cleavable moiety. and the second cleavable moiety is a chemically cleavable moiety. or the first cleavable moiety can be a chemically cleavable moiety, and the second The two cleavable moieties can be enzymatically cleavable moieties. , the first cleavable moiety is a first enzymatically cleavable moiety, and the second cleavable The moiety is a second enzymatically cleavable moiety.

[0191] For example, the cleavable moiety can be a chemically cleavable moiety. Possible moieties include cleavable moieties that can be cleaved in the presence of certain chemical conditions. In some cases, chemically cleavable moieties can be cleaved under certain chemical conditions. dissociates in the presence of a condition to separate the cleavable moiety into two or more separable moieties; For example, a chemically cleavable moiety includes one or more bonds that can be cleaved by chemical modification. Chemical conditions that can lead to hydrolysis of the chemically cleavable moiety, such as acidic conditions or alkali conditions, In some cases, the chemically cleavable moiety can be cleaved in the presence of potassium conditions. The chemical conditions under which the cleavage occurs are determined by the chemical reaction between the desired target site of action, e.g., antibody-drug conjugate and the cleavage site of the cleavage site. In some cases, the drug may be released from the hydroxybenzoate at the desired target site of action. In other words, the chemical conditions found at the desired cleavage site of the chemically cleavable moiety are different from those found at other It is not present in significant amounts in compartments such as whole blood, plasma, or serum. Cleavage of the cleavable moiety occurs at the desired site of action while substantial cleavage occurs at the desired site of action. antibody-drug conjugates reach the desired site of action without significant activity in other compartments It can be controlled so that it does not occur significantly before.

[0192] In some cases, the cleavable moiety may be an enzymatically cleavable moiety. The cleavable moiety can be split into two or more parts as described above through the enzymatic action of an enzyme. An enzymatically cleavable moiety is a cleavable moiety that can be separated from the target protein by an enzyme. Any cleavable moiety that can be cleaved through action, including but not limited to: For example, peptides and glycosides. In some cases, enzymatic cleavage is possible. The enzyme cleaving the available moiety will then target the desired site of action, e.g., antibody-drug conjugate. In some cases, the enzyme is present at the desired target site of action of the drug to be released from the The enzyme that cleaves the specifically cleavable moiety is present in another compartment, e.g., whole blood, plasma, or serum. Therefore, cleavage of the enzymatically cleavable moiety is not expected to result in substantial cleavage. While cleavage occurs at the desired site of action, cleavage does not occur significantly in other compartments, allowing antibody-drug interaction. The conjugate is controlled so that it does not significantly occur before it reaches the desired site of action. This can be done.

[0193] For example, as described herein, the antibody-drug conjugates of the present disclosure can be used to treat cancer For therapy, e.g., for delivery of cancer therapeutic agents to a desired site of action where cancer cells are present. In some cases, enzymes, such as protease enzymes, cathepsins, can be used. B can be a cancer biomarker that is overexpressed in cancer cells. The overexpression, and therefore localization, of certain enzymes in the desired site of action (i.e. i.e., to specifically release drugs at the site of the cancer (and the overexpressed enzyme) The enzymatically cleavable linker included in the cleavable linker of the antibody-drug conjugate of the present disclosure Thus, in some embodiments, the term "enzymatic" can be used in the context of a moiety such as The automatically cleavable portion can be cleaved by an enzyme that is overexpressed in cancer cells. For example, the enzyme may be a protease, a cathepsin, or a cleavable moiety (e.g., a peptide) that can be cleaved. Therefore, in some cases, the enzymatically cleavable moiety can be Prosin B. A cleavable moiety (e.g., a cleavable moiety) that can be cleaved by a protease enzyme, e.g., cathepsin B For example, peptides).

[0194] In certain embodiments, the enzymatically cleavable moiety is a peptide. Suitable for use in cleavable linkers and cleaved through the enzymatic action of an enzyme The peptide can be any peptide that can be used as an enzymatically cleavable moiety. Non-limiting examples of peptides that can be used include, for example, Val-Ala and For example, the first cleavable moiety (i.e., the first cleavable moiety that is protected from premature cleavage by the cleavable moiety of 2) comprises a peptide The presence of an uncleaved second cleavable moiety can be used to inhibit the first cleavable moiety. (peptides) from cleavage by protease enzymes (e.g., cathepsin B) to provide an anti- from the antibody until the body-drug conjugate is at or near the desired target site of action of the drug. In some cases, the premature release of the drug can be substantially reduced or prevented. One of the amino acid residues of the peptide constituting the first cleavable portion comprises a substituent, comprises a second cleavable moiety. In some cases, the second cleavable moiety comprises a glycoside. In other embodiments, the second cleavable moiety comprises a peptide. In an embodiment, the first cleavable moiety comprises a peptide and the second cleavable moiety is a peptide. Includes chid.

[0195] In some embodiments, the enzymatically cleavable moiety is a sugar moiety, e.g., a glycoside (or In some cases, the glycoside is a cleavable glycoside that does not contain a glycoside. This can facilitate an increase in the hydrophilicity of the cleavable linker compared to the linker. Cosides are suitable for use in cleavable linkers and inhibit the enzymatic action of enzymes. The glycoside can be any glycoside that can be cleaved through a second cleavage. a cleavable portion (i.e., a cleavable portion that protects the first cleavable portion from premature cleavage) In other embodiments, the first cleavable moiety can be a glycoside. In other embodiments, the first cleavable moiety comprises a glycoside. In other embodiments, the first cleavable moiety comprises a glycoside, and the second cleavable moiety comprises a glycoside. The cleavable moiety comprises a peptide and the second cleavable moiety comprises a glycoside. wherein the first cleavable moiety comprises a glycoside and the second cleavable moiety comprises a peptide Includes:

[0196] The glycoside is attached to the cleavable linker through a glycosidic bond (covalent bond). Glycosidic bonds can be of various types, including but not limited to: For example, O-glycosidic bonds (O-glycosides), N-glycosidic bonds (glycosylamines), ), S-glycosidic bond (thioglycosidic), or C-glycosidic bond (C-glycosidic) Linking the glycoside to a cleavable linker via a C-glycosyl or C-glycosyl In some cases, the glycosidic bond is an O-glycosidic bond (O-glycoside). In some cases, glycosides are formed enzymatically (e.g., by enzymatic cleavage of glycosidic bonds). is cleaved from the cleavable linker to which it is attached (through mediated hydrolysis) The glycoside can be a glycoside that attaches the glycoside to a cleavable linker. Any convenient enzyme-cleavable linker capable of effecting bond cleavage (hydrolysis) may be used. The glycoside can be removed or cleaved from the linker. of enzymes that can be used to mediate the cleavage (hydrolysis) of glycosidic bonds that attach An example is β-glucuronidase. Other suitable enzymes can also be used to mediate cleavage (hydrolysis) of the hydroxylic acid bond. In some cases, cleavage of the glycosidic bond that attaches the glycoside to the cleavable linker occurs. The enzyme used to mediate the cleavage (hydrolysis) of the drug in the antibody-drug conjugate The enzyme is found at or near the desired site of action. For example, the enzyme is attached to the antibody-drug conjugate. is a lysosomal enzyme found in cells at or near the desired site of action of the drug. In some cases, the enzyme may be an enzyme that mediates cleavage of the first cleavable portion. is an enzyme found at or near the target site where

[0197] In certain embodiments, the first cleavable moiety is an enzymatically cleavable moiety.

[0198] The moiety of interest (e.g., a drug or active agent) can be attached to any of the polypeptides (e.g., antibodies). The polypeptide can be conjugated to any desired site. For example, modifications having a conjugated moiety at or near the C-terminus of a polypeptide. In another example, a polypeptide having a nucleotide sequence at or near the N-terminus of the polypeptide is provided. Examples include modified polypeptides having a conjugated moiety at position Also, a position between the C-terminus and N-terminus of a polypeptide (e.g., an internal site of a polypeptide) ) conjugated moieties. Combinations of the above are also possible, where the peptide is conjugated to two or more moieties. do.

[0199] In certain embodiments, the conjugates of the present disclosure comprise a carboxyl group at the alpha carbon of the amino acid residue. It includes a drug or active agent conjugated to an amino acid residue of a polypeptide. The gate is a gate that determines whether the side chains of one or more amino acid residues in the polypeptide bind to the drug or active agent. attached (e.g., via a linker as described herein) to a drug or active ingredient For example, the conjugate may be The alpha carbon of one or more amino acid residues in the polypeptide is attached to maytansine. maytansine (e.g., attached to maytansine via a linker as described herein) In other cases, the conjugate comprises a polypeptide that has been modified to The α-carbon of one or more amino acid residues in the peptide is attached to the auristatin ( For example, a linker as described herein may be attached to the auristatin. In other cases, the conjugate includes a polypeptide modified to The α-carbon of one or more amino acid residues in the For example, a compound attached to a duocarmycin through a linker as described herein The term "polypeptides" includes polypeptides that have been modified to be soluble in water.

[0200] Embodiments of the present disclosure include polypeptides that are composed of one or more parts, e.g., two parts, three parts Parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, or a conjugate conjugated to 10 or more moieties. The peptide may be conjugated to the polypeptide at one or more sites in the peptide. For example, One or more moieties may be conjugated to a single amino acid residue of the polypeptide. In the case of , one moiety is conjugated to an amino acid residue of the polypeptide. Alternatively, the two moieties may be conjugated to the same amino acid residue in the polypeptide. wherein the first moiety is conjugated to a first amino acid residue of the polypeptide, and the second moiety is is conjugated to a second amino acid residue of the polypeptide. For example, conjugated to a first moiety at an amino acid residue and two at a second amino acid residue Combinations of the above conjugated to other moieties are also possible. Other combinations, limited to: is not conjugated to the first and second moieties, for example, at the first amino acid residue, and a polypeptide conjugated at the second amino acid residue to the third and fourth moieties, etc. is also possible.

[0201] One or more amino acid residues of the polypeptide conjugated to one or more moieties may be naturally occurring. The amino acids may be naturally occurring amino acids, unnatural amino acids, or a combination thereof. A conjugate may also include a moiety conjugated to a naturally occurring amino acid residue of a polypeptide. In other cases, the conjugate is coupled to a non-natural amino acid residue in the polypeptide. The one or more moieties may comprise a single natural or unnatural amino acid as described above. The polypeptide may be conjugated to one or more amino acid residues. Natural or unnatural amino acid residues may be covalently linked to one or more moieties as described herein. For example, two (or more) amino acid residues in a polypeptide may be Each of the groups (e.g., natural or unnatural amino acid residues) may be present at multiple sites in the polypeptide. It may be conjugated to one or two moieties such that the position is modified.

[0202] As described herein, the polypeptide may be conjugated to one or more moieties. In certain embodiments, the moiety of interest is a chemical entity, e.g., a drug or a detectable For example, a drug (e.g., maytansine, auristatin, or duo Calmycin) may be conjugated to the polypeptide, or in other embodiments, a detectable A label capable of binding to the polypeptide may be conjugated to the polypeptide. Conjugates of polypeptides and drugs; Conjugates of polypeptides and detectable Conjugates of two or more drugs and polypeptides and conjugates of two or more detectable labels and polypeptides. Examples of suitable ion exchangers include, but are not limited to, ion exchangers.

[0203] In certain embodiments, the polypeptide and the moiety of interest are coupled via a coupling moiety For example, the polypeptide and the moiety of interest are each coupled through The polypeptide and the target molecule may be linked (e.g., covalently linked) to a binding moiety, thereby Indirectly coupling a moiety of interest (e.g., a drug, e.g., maytansine) through a coupling moiety In some cases, the coupling moiety may be a hydrazinyl-indolyl hydrazinyl-pyrrolo-pyridinyl compounds, or hydrazinyl-indolyl compounds or derivatives of hydrazinyl-pyrrolo-pyridinyl compounds. of interest through indolyl or hydrazinyl-pyrrolo-pyridinyl coupling moieties General scheme for coupling a moiety of interest (e.g., maytansine) to a polypeptide The reaction scheme is shown in the following general reaction scheme: The pyrrolo-pyridinyl coupling moiety is referred to herein as hydrazino-iso-pyridinyl. Tessprenger (HIPS) coupling moiety and aza-hydrazino-iso-pic They are sometimes called Tesprengler (azaHIPS) coupling moieties. . [ka]

[0204] In the above reaction scheme, R represents a group conjugated to a polypeptide (e.g., a group described herein). a moiety of interest (coupled to the polypeptide via a cleavable linker such as those described in As shown in the reaction scheme above, 2-formyl A polypeptide containing a fGly residue (fGly) is coupled to a coupling moiety (e.g., hydra containing hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl coupling moieties The polymer attached to the coupling moiety is reacted with a drug or active agent modified as described above. A polypeptide conjugate is generated, so that the drug or active ingredient can be coupled through the coupling moiety. The activity agent is attached to the polypeptide.

[0205] As described herein, a moiety may be any of a variety of moieties, including but not limited to: For example, it can be a chemical entity, such as a detectable label, or a drug. and R″ are each independently any desired substituent, including but not limited to: Hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl , substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino No, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, amino Noacyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, Substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and Z may be CR 21 , N.R. 22 , N, O or S may be R 21 and R 22 is described above for R' and R'' are each independently selected from the optional substituents.

[0206] As shown in the conjugates and compounds described herein, other hydrazinyl- Indolyl or hydrazinyl-pyrrolo-pyridinyl coupling moieties are also possible For example, hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl couples The linking moiety is attached (e.g., covalently attached) to the linker. Therefore, embodiments of the present disclosure can be used to link drugs or active agents through linkers. hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl caprylate attached to Contains a coupling moiety. Hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridyl Various embodiments of linkers capable of coupling a dianyl coupling moiety to a drug or active agent. These embodiments are described in detail herein. For example, in some cases, the linker may be cleavable. The linker may be a cleavable linker, for example, a cleavable linker as described herein.

[0207] In certain embodiments, the modified polypeptide prior to conjugation to the moiety of interest is Modifications of the polypeptide may be made for conjugation to a moiety of interest. Modified polypeptides can be produced that contain one or more reactive groups suitable for the purpose of In the case of a moiety, the polypeptide is coupled to a moiety of interest (e.g., a coupling moiety, e.g., Hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl cups as described above to provide one or more reactive groups suitable for conjugation to a moiety (including a ring moiety) For example, the polypeptide may be modified at one or more amino acid residues. It may be modified to include a reactive aldehyde group (e.g., a reactive aldehyde). The aldehyde may be contained in an "aldehyde tag" or "ald-tag" "aldehyde tag" or "ald-tag" as used herein refers to a 2-formyl Formyl to contain a glycine residue (referred to herein as "FGly"). The sulfatase motif converted by the action of glycine generating enzyme (FGE) (e.g. The amino acid sequence derived from FGl (L(C / S)TPSR) is The γ residue is sometimes called "formylglycine." In other words, it is an "aldehyde group." The term "sulfatase motif" refers to a "converted" sulfatase motif (i.e., a cysteine ​​or The serine residue is converted to FGly by the action of FGE, e.g., a sulfatase motif. For example, L(FGly)TPSR. The converted sulfatase motif is a nucleotide sequence that is significantly different from the "unconverted" sulfatase motif. - (i.e., the cysteine ​​or serine residue is not converted to FGly by FGE) However, sulfatase motifs that can be converted, such as the sequence: L(C / S)T It is derived from an amino acid sequence containing an unconverted sulfatase motif with PSR. The action of formylglycine generating enzyme (FGE) on the sulfatase motif "Conversion" as used in this context refers to the conversion of a sulfa to a formylglycine (FGly) residue. Biochemical modification of cysteine ​​or serine residues in the tRNA motif (e.g., Cys to FG) ly or Ser to FGly). Aldehyde tag and site-specific protein Additional aspects of its use in protein modification are described in U.S. Pat. No. 7,985,783 and U.S. Pat. and U.S. Pat. Nos. 8,729,232, the disclosures of each of which are incorporated herein by reference. It is incorporated herein by reference.

[0208] In some cases, modified polypeptides containing FGly residues are reacted with compounds (e.g., hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl compounds as described above Conjugation to the moiety of interest by reaction of FGly with a compound containing a coupling moiety For example, a FGly-containing polypeptide may be combined with a drug containing a reactive partner. The contacting may be carried out under conditions suitable to provide for conjugation of the drug to the peptide. The reactive partner-containing drug may be a hydrazinyl-indolyl or hydrazinyl-indolyl as described above. It may also contain a pyridinyl-pyrrolo-pyridinyl coupling moiety, e.g., a drug or active ingredient. The agent is a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl coupling agent. In some cases, the drug or active agent may be modified to include a hydrazine moiety. attached to hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl, e.g., linker, for example, a hydra linker via a cleavable linker as described in detail herein. Covalently attached to hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl can be.

[0209] In certain embodiments, the conjugates of the present disclosure comprise at least one modified amino acid. The amino acid residues of the polypeptides are also included (e.g., antibodies). The amino acid residue may be hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyrrolyl as described above. It may be coupled to a drug or active agent containing a lysinyl coupling moiety. In certain embodiments, the modified amino acid residues of the polypeptide (e.g., antibody) are It may also be derived from a cysteine ​​or serine residue that has been converted to a FGly residue as described above. In certain embodiments, the FGly residue is a hydrazinyl-indolyl or Conjugated to a drug or active agent containing a hydrazinyl-pyrrolo-pyridinyl coupling moiety and the drug or active agent is hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyrazole. Conjugates of the present disclosure are provided that are coupled to a polypeptide via a divinyl coupling moiety. As used herein, the term FGly' refers to a moiety of interest (e.g., Modification of a polypeptide (e.g., an antibody) that is coupled to a target polypeptide (e.g., a drug or active agent) It refers to the amino acid residues that are

[0210] In certain embodiments, the conjugate comprises at least one of the following: and one modified amino acid residue, the modified amino acid residue being one of those described herein. The linker is then attached to a linker (cleavable linker) such that the drug For example, the conjugate may be at least one of the above-described In some embodiments, each of the amino acid residues may contain one modified amino acid residue (FGly'). , the conjugate has the formula (I): [ka]

[0211] is a conjugate of Z is CR 4 or N, R 1 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, Substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl and substituted heterocyclyl; R 2 and R 3 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, a amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy , acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl aryl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, hetero aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; is R 2 and R 3 may be cyclically linked to form a 5- or 6-membered heterocyclyl. Ku, Each R 4 is hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, Amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy Acylamino, aminoacyl, alkylamido, substituted alkylamido, sulfo aryl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, hydroxyl Heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl , heterocyclyl, and substituted heterocyclyl; Each R 5 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl , alkynyl, and substituted alkynyl; Each R 6 is an alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkyn ... alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; k is an integer from 1 to 10, R 7 is hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, a amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy , acylamino, aminoacyl, alkylamido, substituted alkylamido, aryl , substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, Selected from substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl And, L 1 is the first linker, L 2 is a second linker, W 1 is a drug, and W 2 is an antibody, L 1 , R 6 or R 7 One of the fragments comprises a second cleavable portion.

[0212] In certain embodiments of Formula (I), n is 2 and the conjugate has the formula (Ia ): [ka]

[0213] is a conjugate of R 6’ or R 6’’ one of which contains a second cleavable moiety, and R 6’ and R 6’’ The other of the groups is alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heteroaryl and substituted heterocyclyl.

[0214] For example, the conjugate may have the formula (Ib): [ka]

[0215] It can be a conjugate of:

[0216] In other instances, the conjugate has the formula (Ic): [ka]

[0217] It can be a conjugate of:

[0218] In certain embodiments of Formula (I), k is 2 and the conjugate has the formula (Id ): [ka]

[0219] wherein: R 6’ and R 6’’ is an alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, hetero Aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heteroaryl independently selected from tetracyclyl, tetracyclyl, and substituted heterocyclyl; and R 7 comprises a second cleavable moiety.

[0220] For example, the conjugate may have the formula (Ie): [ka]

[0221] It can be a conjugate of:

[0222] In certain embodiments of Formula (I), k is 2 and the conjugate has the formula (If ): [ka]

[0223] is a conjugate of R 6’ and R 6’’ is an alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, hetero Aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heteroaryl independently selected from tetracyclyl, tetracyclyl, and substituted heterocyclyl; and R 8 comprises a second cleavable moiety.

[0224] For example, the conjugate may have the formula (Ig): [ka]

[0225] It can be a conjugate of:

[0226] The substituents for the conjugates of formula (I) are described in more detail below. Reference to (I) includes formulas (Ia), (Ib), (Ic), (Id), (Ie), (If ) and (Ig).

[0227] In certain embodiments, Z is CR 4 or N. In certain embodiments, Z is CR 4 In certain embodiments, Z is N.

[0228] In certain embodiments, R 1 is hydrogen, alkyl, substituted alkyl, alkenyl , substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cyclo In particular, the alkyl group is selected from alkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R 1 is hydrogen. In certain embodiments, R 1 is alkyl or Substituted alkyl, e.g., C 1~6 Alkyl or C 1~6 Substituted alkyl of , or C 1~4 Alkyl or C 1~4 substituted alkyl of, or C 1~3 Al Kill or C 1~3 In certain embodiments, R 1 Hame In certain embodiments, R 1 is alkenyl or substituted alkenyl, For example, C 2~6 Alkenyl or C 2~6 substituted alkenyl of C 2~4 Alkenyl or C 2~4 substituted alkenyl of C 2~3 Alkenyl or is C 2~3 In certain embodiments, R 1 is alkynyl or substituted alkynyl, e.g., C 2~6 Alkenyl or C 2~6 is replaced by alkenyl, or C 2~4 Alkenyl or C 2~4 substituted alkenyl, or or C 2~3Alkenyl or C 2~3 In certain embodiments, the alkenyl group is a substituted alkenyl. In the embodiment, R 1 is aryl or substituted aryl, e.g., C 5~8 Aryl also is C 5~8 substituted aryl, for example, C5 aryl or C5 substituted aryl C6 aryl, or C6 aryl or C6 substituted aryl. In this state, R 1 is heteroaryl or substituted heteroaryl, e.g., C 5~8 Haitai Roaryl or C 5~8 Substituted heteroaryls such as C5 heteroaryls are also or C5 substituted heteroaryl, or C6 heteroaryl or C6 substituted In certain embodiments, R 1 is cycloalkyl or substituted cycloalkyl, e.g., C 3~8 Cycloalkyl or C 3~8 substituted Cycloalkyl, e.g., C 3~6 Cycloalkyl or C 3~6 Substituted cyclo Alkyl, or C 3~5 Cycloalkyl or C 3~5 Substituted cycloalkyl of In certain embodiments, R 1 is heterocyclyl or substituted heterocyclyl For example, C 3~8 Heterocyclyl or C 3~8 substituted heterocyclyl, e.g. Ba, C 3~6 Heterocyclyl or C 3~6 substituted heterocyclyl, or C3 ~5 Heterocyclyl or C 3~5 is a substituted heterocyclyl of the formula:

[0229] In certain embodiments, R 2 and R 3 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, Substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester aryl, acyl, acyloxy, acylamino, aminoacyl, alkylamido, substituted Alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, Substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, respectively; are independently selected, or R 2 and R 3 are cyclically linked to form a 5- or 6-membered heterocycle. It may also form krill.

[0230] In certain embodiments, R 2 is hydrogen, alkyl, substituted alkyl, alkenyl , substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted Alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl , acyloxy, acylamino, aminoacyl, alkylamido, substituted alkyla amido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted Aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl and is selected from chloroalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R 2 is hydrogen. In certain embodiments, R 2 is alkyl or is a substituted alkyl, e.g., C 1~6 Alkyl or C 1~6 Substituted alkyl Le, or C 1~4 Alkyl or C 1~4 substituted alkyl of, or C 1~3 a Rukill or C 1~3 In certain embodiments, R 2 teeth In certain embodiments, R 2 is alkenyl or substituted alkenyl , e.g., C 2~6 Alkenyl or C 2~6 substituted alkenyl of C 2~ 4Alkenyl or C 2~4 substituted alkenyl of C 2~3 If alkenyl Kuha C 2~3 In certain embodiments, R 2 Alkini In certain embodiments, R is aryl or substituted alkynyl. 2 is alkoxy or is substituted alkoxy. In certain embodiments, R 2 is amino or substituted In certain embodiments, R 2 is carboxyl or carboxyl ester In certain embodiments, R 2 is acyl or acyloxy. In certain embodiments, R 2 is acylamino or aminoacyl. So, R 2 is an alkylamide or a substituted alkylamide. In this state, R 2 is sulfonyl. In certain embodiments, R 2 is thioalkoxy or is a substituted thioalkoxy. In certain embodiments, R 2 is aryl or substituted Substituted aryl, e.g., C 5~8 Aryl or C 5~8 Substituted aryl of, e.g. For example, C5 aryl or C5 substituted aryl, or C6 aryl or C 6-substituted aryl. In certain embodiments, R 2 is heteroaryl or Substituted heteroaryl, e.g., C 5~8 Heteroaryl or C 5~8 is replaced by heteroaryl, for example, C5 heteroaryl or C5 substituted heteroaryl C6 heteroaryl or C6 substituted heteroaryl. In certain embodiments, R 2 is cycloalkyl or substituted cycloalkyl, e.g., C 3~8 Cycloalkyl or C 3~8 substituted cycloalkyl, such as C 3~6 S Chloroalkyl or C 3~6 substituted cycloalkyl of C 3~5 Cycloal Kill or C 3~5 In certain embodiments, R 2 is heterocyclyl or substituted heterocyclyl, e.g., C 3~6 Heterocyclyl Or C 3~6 substituted heterocyclyl of C 3~5 Heterocyclyl or C 3~5 is a substituted heterocyclyl of the formula:

[0231] In certain embodiments, R 3 is hydrogen, alkyl, substituted alkyl, alkenyl , substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted Alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl , acyloxy, acylamino, aminoacyl, alkylamido, substituted alkyla amido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted Aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl and is selected from chloroalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R 3 is hydrogen. In certain embodiments, R 3 is alkyl or is a substituted alkyl, e.g., C 1~6 Alkyl or C 1~6 Substituted alkyl Le, or C 1~4 Alkyl or C 1~4 substituted alkyl of, or C 1~3 a Rukill or C 1~3 In certain embodiments, R 3 teeth In certain embodiments, R 3 is alkenyl or substituted alkenyl , e.g., C 2~6 Alkenyl or C 2~6 substituted alkenyl of C 2~ 4Alkenyl or C 2~4 substituted alkenyl of C 2~3 If alkenyl Kuha C 2~3 In certain embodiments, R 3 Alkini In certain embodiments, R is aryl or substituted alkynyl. 3 is alkoxy or is substituted alkoxy. In certain embodiments, R 3 is amino or substituted In certain embodiments, R 3 is carboxyl or carboxyl ester In certain embodiments, R 3 is acyl or acyloxy. In certain embodiments, R 3 is acylamino or aminoacyl. So, R 3 is an alkylamide or a substituted alkylamide. In this state, R 3 is sulfonyl. In certain embodiments, R 3 is thioalkoxy or is a substituted thioalkoxy. In certain embodiments, R 3 is aryl or substituted Substituted aryl, e.g., C 5~8 Aryl or C 5~8 Substituted aryl of, e.g. For example, C5 aryl or C5 substituted aryl, or C6 aryl or C 6-substituted aryl. In certain embodiments, R 3 is heteroaryl or Substituted heteroaryl, e.g., C 5~8 Heteroaryl or C 5~8 is replaced by heteroaryl, for example, C5 heteroaryl or C5 substituted heteroaryl C6 heteroaryl or C6 substituted heteroaryl. In certain embodiments, R 3 is cycloalkyl or substituted cycloalkyl, e.g., C 3~8 Cycloalkyl or C 3~8 substituted cycloalkyl, such as C 3~6 S Chloroalkyl or C3~6 substituted cycloalkyl of C 3~5 Cycloal Kill or C 3~5 In certain embodiments, R 3 is heterocyclyl or substituted heterocyclyl, e.g., C 3~8 Heterocyclyl or C 3~8 Substituted heterocyclyl, for example, C 3~6 Heterocyclyl or C 3~6 substituted heterocyclyl of C 3~5 Heterocyclyl or C 3~5 is a substituted heterocyclyl of the formula:

[0232] In certain embodiments, R 2 and R 3 is cyclically linked to form a 5- or 6-membered heterocycle In certain embodiments, R 2 and R 3 The number of rings is 5. or a 6-membered heterocyclyl. In certain embodiments, R 2 and R 3 teeth In certain embodiments, R 2 Oh BiR 3 are cyclically linked to form a 6-membered heterocyclyl.

[0233] In certain embodiments, each R 4 is hydrogen, halogen, alkyl, substituted alkyl , alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy , substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester Ter, acyl, acyloxy, acylamino, aminoacyl, alkylamido, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl , substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, Independently of substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl and selected.

[0234] Each R 4 The various possibilities for are described in more detail below. In terms of form, R 4 is hydrogen. In certain embodiments, each R 4 is hydrogen. In an embodiment of the present invention, R 4 is a halogen, for example, F, Cl, Br or I. In embodiments, R 4 is F. In certain embodiments, R 4 is Cl. In an embodiment of the present invention, R 4 is Br. In certain embodiments, R 4 is I. In certain embodiments, R 4 is alkyl or substituted alkyl, e.g., C 1~6 Archi Lu or C 1~6 substituted alkyl of, or C 1~4 Alkyl or C 1~4 of substituted alkyl, or C 1~3 Alkyl or C 1~3 With substituted alkyl In certain embodiments, R 4 is methyl. In certain embodiments, R 4 teeth Alkenyl or substituted alkenyl, e.g., C 2~6 Alkenyl or C 2~6 substituted alkenyl of C 2~4 Alkenyl or C2~4 Substituted Al Kenyl, or C 2~3 Alkenyl or C 2~3 It is a substituted alkenyl. In certain embodiments, R 4 is alkynyl or substituted alkynyl. In an embodiment of the present invention, R 4 is alkoxy or substituted alkoxy. In terms of form, R 4 is amino or substituted amino. In certain embodiments, R 4 is carboxyl or carboxyl ester. In certain embodiments, R 4 Haa In certain embodiments, R 4 is acylamino or acylamino In certain embodiments, R 4 is an alkylamide or a substituted alkyl In certain embodiments, R 4 is a sulfonyl. In the embodiment, R 4 is a thioalkoxy or substituted thioalkoxy. In embodiments, R 4 is aryl or substituted aryl, e.g., C 5~8 Arilma or C 5~8 substituted aryl, for example, C5 aryl or C5 substituted aryl C6 aryl or C6 substituted aryl (e.g., phenyl or In certain embodiments, R 4 Heteroaryl or is a substituted heteroaryl, e.g., C 5~8 Heteroaryl or C 5~8 Substitution of C5 heteroaryl or C5 substituted heteroaryl, for example C6 heteroaryl or C6 substituted heteroaryl. In certain embodiments, R 4 is cycloalkyl or substituted cycloalkyl, for example, C 3~8 Cycloalkyl or C 3~8 substituted cycloalkyl, such as C 3~6 Cycloalkyl or C 3~6 substituted cycloalkyl of C 3~5 Cycloa Rukill or C 3~5 In certain embodiments, R 4 is heterocyclyl or substituted heterocyclyl, e.g., C 3~8 Heterocycly Le or C 3~8 Substituted heterocyclyl, for example, C 3~6 Heterocyclyl or is C 3~6 substituted heterocyclyl of C 3~5 Heterocyclyl or C 3~ 5-substituted heterocyclyl.

[0235] In certain embodiments, each R 5 is hydrogen, alkyl, substituted alkyl, alkenyl independently selected from alkyl, substituted alkenyl, alkynyl, and substituted alkynyl; In certain embodiments, R 5 is hydrogen. In certain embodiments, R 5 teeth Alkyl or substituted alkyl, e.g., C 1~6 Alkyl or C 1~6 Substitution of alkyl, or C 1~4 Alkyl or C 1~4substituted alkyl of, is C 1~3 Alkyl or C 1~3 In certain embodiments, So, R 5 is methyl. In certain embodiments, R 5 is ethyl. In embodiments, R 5 is propyl (e.g., n-propyl or isopropyl). In certain embodiments, R 5 is butyl (e.g., n-butyl, isobutyl, sec-butyl) In certain embodiments, R 5 is pentyl (e.g., n -pentyl or neopentyl). In certain embodiments, R 5 is Neopen In certain embodiments, R 5 is alkenyl or substituted alkenyl, For example, C 2~6 Alkenyl or C 2~6 substituted alkenyl of C 2~4 Alkenyl or C 2~4 substituted alkenyl of C 2~3 Alkenyl or is C 2~3 In certain embodiments, R 5 is alkynyl or substituted alkynyl.

[0236] In certain embodiments, each R 6 is an alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloaryl and each independently selected from alkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R 6 is hydrogen. In certain embodiments, R 6 Is alkyl or substituted alkyl, e.g., C 1~6 Alkyl or C 1~6 Substituted Al Kill, or C 1~4 Alkyl or C 1~4 substituted alkyl of, or C 1~3 Alkyl or C 1~3 In certain embodiments, R 6 is methyl. In certain embodiments, R 6 is ethyl. is R 6 is propyl (e.g., n-propyl or isopropyl). In the embodiment, R 6 is butyl (e.g., n-butyl, isobutyl, sec-butyl, or t-butyl). In certain embodiments, R 6 is pentyl (e.g., n-pentyl or neopentyl). In certain embodiments, R 6 is neopentyl In certain embodiments, R 6 is alkenyl or substituted alkenyl, e.g., C 2~6 Alkenyl or C 2~6 substituted alkenyl of C 2~4 Alkenyl Or C 2~4 substituted alkenyl of C 2~3 Alkenyl or C 2~3 In certain embodiments, R 6 is alkynyl or substituted It is an alkynyl group.

[0237] In certain embodiments, R 6 is aryl or substituted aryl, e.g., C 5~ 8 aryl or C 5~8 substituted aryl, for example, C5 aryl or C5 Substituted aryl, or C6 aryl or C6 substituted aryl (e.g., phenyl or substituted phenyl). In certain embodiments, R 6 is hetero Aryl or substituted heteroaryl, e.g., C 5~8 Heteroaryl or C5 ~8 substituted heteroaryl, for example, C5 heteroaryl or C5 substituted heteroaryl heteroaryl, or C6 heteroaryl or C6 substituted heteroaryl In certain embodiments, R 6 is cycloalkyl or substituted cycloalkyl For example, C 3~8 Cycloalkyl or C 3~8 substituted cycloalkyl, e.g. Ba, C 3~6 Cycloalkyl or C 3~6 substituted cycloalkyl, or C3 ~5 Cycloalkyl or C 3~5 In certain embodiments, the substituted cycloalkyl is In the embodiment, R 6 is heterocyclyl or substituted heterocyclyl, e.g., C 3~8 Heterocyclyl or C 3~8 Substituted heterocyclyl, for example, C 3~6 Heterogeneity Krill or C 3~6 substituted heterocyclyl of C 3~5 Heterocyclyl Or C 3~5 is a substituted heterocyclyl of the formula:

[0238] In certain embodiments, R 6 represents the side chain of an amino acid. For example, R 6 Amy is heaven The α-carbon of amino acid residues, including natural amino acids, unnatural amino acids, and amino acid analogs, In some cases, R 6 is found in naturally occurring proteins The side chains of the amino acids found (e.g., Ala or A, Cys or C, Asp or D) , Glu or E, Phe or F, Gly or G, His or H, Ile or I , Lys or K, Leu or L, Met or M, Asn or N, Pro or P , Gln or Q, Arg or R, Ser or S, Thr or T, Val or V , Trp or W, Tyr or Y). 6 Haba represents the side chain of phosphorus (Val), i.e., R 6 is isopropyl. In this state, R 6 represents the side chain of alanine (Ala), i.e., R 6 is methyl. In certain embodiments, R 6 represents the side chain of phenylalanine (Phe), i.e., R 6 is benzyl. In certain embodiments, R 6 represents the side chain of lysine (Lys), That is, R 6 is 4-amino-butyl.

[0239] In certain embodiments, k is an integer from 1 to 10. In certain embodiments, k is In certain embodiments, k is 1. In certain embodiments, k is 2. In certain embodiments, k is 3. In certain embodiments, k is 4. In certain embodiments, k is 5. In certain embodiments, k is 6. In certain embodiments, k is 7. In certain embodiments, k is 8. In certain embodiments, k is 9. In certain embodiments, k is 10.

[0240] In certain embodiments, R 7 is hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, Substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester aryl, acyl, acyloxy, acylamino, aminoacyl, alkylamido, substituted Alkylamide, aryl, substituted aryl, heteroaryl, substituted heteroaryl aryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted and heterocyclyl.

[0241] In certain embodiments, R 7 is hydrogen. In certain embodiments, R 7 Ha Haroge For example, F, Cl, Br, or I. In certain embodiments, R 7 is alkyl or substituted alkyl, e.g., C 1~6 Alkyl or C 1~6 The replaced Rukill, or C 1~4 Alkyl or C 1~4 substituted alkyl of, or C 1~ 3 alkyl or C 1~3 In certain embodiments, R 7 is methyl. In certain embodiments, R 7 is ethyl. So, R 7is propyl (e.g., n-propyl or isopropyl). In embodiments, R 7 is butyl (e.g., n-butyl, isobutyl, sec-butyl, or is t-butyl). In certain embodiments, R 7 is pentyl (e.g., n-pentyl) In certain embodiments, R 7 is neopentyl In certain embodiments, R 7 is alkenyl or substituted alkenyl, for example, C 2~6 Alkenyl or C 2~6 substituted alkenyl of C 2~4 Arkeny Lu or C 2~4 substituted alkenyl of C 2~3 Alkenyl or C 2~ In certain embodiments, R 7 is alkynyl or and substituted alkynyl.

[0242] In certain embodiments, R 7 is alkoxy or substituted alkoxy. In certain embodiments, R 7 is amino or substituted amino. So, R 7 is carboxyl or carboxyl ester. In certain embodiments, , R 7 is acyl. In certain embodiments, R 7 is acyloxy. In an embodiment of the present invention, R 7 is acylamino. In certain embodiments, R 7 Aminoa In certain embodiments, R 7 is an alkylamide or a substituted alkyl It is an amide.

[0243] In certain embodiments, R 7 is aryl or substituted aryl, e.g., C 5~ 8 aryl or C 5~8 substituted aryl, for example, C5 aryl or C5 Substituted aryl, or C6 aryl or C6 substituted aryl (e.g., phenyl or substituted phenyl). In certain embodiments, R 7 is hetero Aryl or substituted heteroaryl, e.g., C 5~8 Heteroaryl or C5 ~8 substituted heteroaryl, for example, C5 heteroaryl or C5 substituted heteroaryl heteroaryl, or C6 heteroaryl or C6 substituted heteroaryl In certain embodiments, R 7 is cycloalkyl or substituted cycloalkyl For example, C 3~8 Cycloalkyl or C 3~8 substituted cycloalkyl, e.g. Ba, C 3~6 Cycloalkyl or C 3~6 substituted cycloalkyl, or C3 ~5 Cycloalkyl or C 3~5 In certain embodiments, the substituted cycloalkyl is In the embodiment, R 7 is heterocyclyl or substituted heterocyclyl, e.g., C 3~8 Heterocyclyl or C 3~8 Substituted heterocyclyl, for example, C 3~6 Heterogeneity Krill or C 3~6 substituted heterocyclyl of C 3~5 Heterocyclyl Or C 3~5 is a substituted heterocyclyl of the formula:

[0244] In certain embodiments, L 1 is the first linker. L 1 Suitable linkers for is described in more detail below.

[0245] In certain embodiments, L 2 is the second linker. 2 Suitable linkers for is described in more detail below.

[0246] In certain embodiments, W 1 is a drug. For example, W 1 is a maytansinoid Further description of maytansinoids can be found in the disclosure herein. In some cases, W 1 is an auristatin. Further description of auristatins is provided herein. In some cases, the W 1 is a duocarmycin. Further description of ocarmycins is found in the disclosure herein.

[0247] In certain embodiments, W 2 is an antibody. Further description of such antibodies can be found in the disclosure herein.

[0248] In certain embodiments, the compound of formula (I) comprises one or more linkers. The coupling moiety may be one or more moieties of interest and / or one or more In some embodiments, the linker may be utilized to link multiple polypeptides. The coupling moiety can be used to attach either a polypeptide or a chemical entity, e.g., a drug. The linker may be attached at any convenient position (e.g., as described herein). The linker may be attached (e.g., covalently attached) to the coupling moiety. For example, the linker may be Hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl coupling moieties The hydrazinyl- The indolyl or hydrazinyl-pyrrolo-pyridinyl coupling moiety is a linker ( and therefore drugs) to polypeptides, e.g., antibodies. For example, the coupling moiety can be used to attach the linker (and thus the drug) to the modified polypeptide. may be used to conjugate to a specific amino acid residue, e.g., FGly residue of an antibody. .

[0249] In certain embodiments, the linker comprises one or more linkers, e.g., a first linker Carr, L. 1 , and a second linker L 2 Additionally, the linker may be any of the compounds described herein. one or more cleavable moieties (e.g., a first cleavable moiety and a second cleavable moiety) as described above. In some cases, the linker may comprise one or more linkers. Car, e.g., a first linker, L 1 , and a second linker L 2 For example, phosphorus The car connects the first cleavable moiety to a coupling moiety (e.g., a cleavable moiety as described herein). Suitable hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl coupling moieties a first linker (L 1 ), and the first cleavable moiety is a chemical entity, e.g. For example, a second linker (L) linked to the drug as described herein 2) may also be included Therefore, the linker may be a hydrazinyl-indoline (e.g., as described herein). First cleavable (through the aryl or hydrazinyl-pyrrolo-pyridinyl coupling moiety) a first linker (L) that connects the functional moiety to the antibody; 1 ), and chemically linking the first cleavable moiety a second linker (L) that links the target entity, e.g., a drug as described herein. 2 )of It may include.

[0250] For example, as shown in formula (I) above, L 1 is W through the coupling part 2 to It is attached, so W 2 is connected to a first linker L through a coupling moiety 1 As mentioned above, W 2 is an antibody, so L 1 is a couple is attached to the antibody through a linking moiety, e.g., a first linker L 1 Is a cuplin through a linking moiety (e.g., hydrazinyl-indolyl or Indirectly linked to the antibody (through a hydrazinyl-pyrrolo-pyridinyl coupling moiety) Additionally, as shown in formula (I) above, L 1 is L 2 Attached (indirectly) to It is also L 2 is W 1 As mentioned above, the W 1 is a drug, Therefore, the second linker L 2 is the first linker L 1 and a coupling moiety (e.g. , hydrazinyl-indolyl or hydrazinyl-pyrrolo- as described herein Drugs can be attached to antibodies via the pyridinyl coupling moiety 2 Attach to.

[0251] The first linker (L) in the subject conjugates and compounds 1 ) and a second linker -(L 2 Any convenient linker may be utilized for the is the first linker (L 1 ) and a second linker (L 2 ) are each independently alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carbo xyl, carboxyl ester, acylamino, alkylamide, substituted alkyl Amido, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cyclohexyl Cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl In certain embodiments, the first linker (L 1 ) and a second linker (L 2 ) are each independently alkyl or substituted alkyl. In certain embodiments, the first linker (L 1 ) and the second The linker (L 2 ) each independently represent an alkenyl or substituted alkenyl group; In certain embodiments, the first linker (L 1 ) and a second linker (L 2 ) may each independently comprise an alkynyl or substituted alkynyl group. In certain embodiments, the first linker (L 1 ) and a second linker (L2 )teeth , each independently may comprise an alkoxy or substituted alkoxy group. In certain embodiments, the first linker (L 1 ) and a second linker (L 2 ) are respectively may independently comprise an amino or substituted amino group. The first linker (L 1 ) and a second linker (L 2 ) are each independently a carboxylic acid. In certain embodiments, the first linker may comprise a hydroxyl or carboxyl ester group. L 1 ) and a second linker (L 2 ) each independently represents an acylamino group In certain embodiments, the first linker (L 1 ) and a second linker (L 2 ) each independently contain an alkylamido or substituted alkylamido group. In certain embodiments, the first linker (L 1 ) and a second linker ( L 2 ) may each independently comprise an aryl or substituted aryl group. In certain embodiments, the first linker (L 1 ) and a second linker (L 2 ) is that Each may independently comprise a heteroaryl or substituted heteroaryl group. In certain embodiments, the first linker (L 1 ) and a second linker (L 2 ) are Each may independently comprise a cycloalkyl or substituted cycloalkyl group. In certain embodiments, the first linker (L 1 ) and a second linker (L 2 ) are respectively It may independently comprise a heterocyclyl or substituted heterocyclyl group.

[0252] In certain embodiments, the first linker (L 1 ) and a second linker (L 2 )teeth, Each may independently comprise a polymer. For example, the polymer may be a polyalkylene glycol. and polyalkylene glycol and its derivatives. Examples include polyethylene glycol, methoxypolyethylene glycol, and polyethylene glycol. Cholesterol homopolymer, polypropylene glycol homopolymer, propylene glycol Copolymers of ethylene glycol (e.g., homopolymers and copolymers are unsubstituted) or substituted at one end with an alkyl group), polyvinyl alcohol, poly vinyl ethyl ether, polyvinylpyrrolidone, and combinations thereof. In certain embodiments, the polymer is a polyalkylene glycol. In an embodiment, the polymer is polyethylene glycol. Other linkers are also possible, as shown in the conjugates and compounds shown.

[0253] In some embodiments, L 1 is the formula -(L 11 ) a -(L 12 ) b -(L 13 ) c -(L 14 ) d - is a first linker described by the formula: 11 , L 12 , L 13 Oh BiL 14are each independently a first linker subunit, and a, b, c and Each d is independently 0 or 1, and the sum of a, b, c and d is 1 to 4.

[0254] In certain embodiments, the sum of a, b, c, and d is 1. In certain embodiments, the sum of a, b, c, and d is 2. In certain embodiments, the sum of a, b, c, and d is 4. In certain embodiments, a, b, c, and d are each 1. In certain embodiments, a, b, and c are each 1, and d is 0. In certain embodiments, a and a and b are each 1, and c and d are each 0. where a is 1 and b, c and d are each 0.

[0255] In certain embodiments, L 11 is a hydroxybenzoate (e.g., as shown in formula (I) above). hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl coupling moieties In certain embodiments, L 12 If there is, it is the first cut In certain embodiments, L 13 If it exists, It is attached to the first cleavable portion. In certain embodiments, L 14 There exists If present, it is attached to the first cleavable portion.

[0256] Any convenient linker subunit can be the first linker L 1 May be used in Linker subunits of interest include polymeric units, e.g., polyethylene glycols. Polyol, polyethylene and polyacrylate, amino acid residue(s), carbon hydrate-based polymers or carbohydrate residues and their derivatives, polynucleotides, Alkyl groups, aryl groups, heterocyclic groups, combinations thereof, and substituted groups thereof In some embodiments, L 11 , L 12 , L 13 and L 14 Each of (if present) is polyethylene glycol, modified Polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups Linking groups including alkylene groups, substituted aryl groups, and diamines (e.g., alkylenediamines) )

[0257] In some embodiments, L 11 If present, polyethylene glycol, modified Polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups In some embodiments, L 11 Is Poly In some embodiments, L 11 is a modified polyethylene glycol In some embodiments, L 11 In some embodiments, , L 11 In some embodiments, L 11 teeth In some embodiments, L 11 is diamine (e.g., a linking group comprising an alkylenediamine).

[0258] In some embodiments, L 12 If present, polyethylene glycol, modified Polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups In some embodiments, L 12 Is Poly In some embodiments, L 12 is a modified polyethylene glycol In some embodiments, L 12 In some embodiments, , L 12 In some embodiments, L 12 teeth In some embodiments, L 12 is diamine (e.g., a linking group comprising an alkylenediamine).

[0259] In some embodiments, L 13 If present, polyethylene glycol, modified Polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups In some embodiments, L 13 Is Poly In some embodiments, L 13 is a modified polyethylene glycol In some embodiments, L 13 In some embodiments, , L 13 In some embodiments, L 13 teeth In some embodiments, L 13 is diamine (e.g., a linking group comprising an alkylenediamine).

[0260] In some embodiments, L 14 If present, polyethylene glycol, modified Polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups In some embodiments, L 14 Is Poly In some embodiments, L 14 is a modified polyethylene glycol In some embodiments, L 14 In some embodiments, , L 14 In some embodiments, L 14 teeth In some embodiments, L 14 is diamine (e.g., a linking group comprising an alkylenediamine).

[0261] In some embodiments, L 1 is (L 11 ) a -(L 12 ) b -(L 13 ) c -(L 14 ) d - a first linker comprising: -(L 11 ) a -ha-(T 1 -V 1 ) a - and -(L 12 ) b -ha-(T 2 -V 2 ) b - and -(L 13 ) c -ha-(T 3 -V 3 ) c -and -(L 14 ) d -ha-(T 4 -V 4 ) d - and T 1 , T 2 , T 3 and T 4 When present, they are tether groups, V 1 , V 2 , V 3 and V 4 When present, they are linked by covalent bonds or functional groups. Yes, and a, b, c and d are each independently 0 or 1, and the sum of a, b, c and d is 1 to 4.

[0262] As noted above, in certain embodiments, L 11 (e.g., as shown in formula (I) above) Hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl cuprates (such as Therefore, in one particular embodiment, the T 1 (for example, For example, hydrazinyl-indolyl or hydrazinyl- In certain embodiments, V 1 is attached to the first cleavable portion. In certain embodiments, L 12 but If present, it is attached to the first cleavable portion. In the embodiment, T 2 If present, it is attached to the first detachable portion or V 2 If present, it is attached to the first severable portion. In certain embodiments, L 13If present, it is attached to the first detachable part Therefore, in certain embodiments, T 3 If there is one, it is the first attached to a breakable part or V 3 If there is, it is the first cleavable In certain embodiments, the L 14 If there is a attached to the first cleavable portion. 4 If present, it is attached to the first detachable portion, or V 4 exists If so, it is attached to the first severable portion.

[0263] Tether group, T 1 , T 2 , T 3 and T 4 With respect to any convenient tether group, the subject phosphorus In some embodiments, the T 1 , T 2 , T 3 and T 4 Noso Each of them is (C1~C 12 ) alkyl, substituted (C1-C 12 ) Alkyl, aryl , substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, Substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (ED A) w , (PEG) n , (AA) p , -(CR 13 OH) h -, piperidine-4-amino (4AP), independently selected from acetal groups, disulfides, hydrazines, and esters. wherein w is an integer of 1 to 20, and n is an integer of 1 to 30. where p is an integer of 1 to 20 and h is an integer of 1 to 12.

[0264] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 and / or T 4 ) (C1~C 12 ) alkyl or substituted (C1-C 12 ) alkyl. In certain embodiments, (C1 to C 12 ) Alkyl is an alkyl group having 1 to 12 carbon atoms, e.g., 1 to 10 carbon atoms, or 1 to 8 carbon atoms, or 1 to 6 carbon atoms, or 1 to Straight or branched chains containing 5 carbon atoms, or 1-4 carbon atoms, or 1-3 carbon atoms is a branched alkyl group. In some cases, (C1-C 12 ) alkyl is alkyl or substituted alkyl, e.g., C1-C 12 Alkyl, or C1-C 10 Archi It may be alkyl, C1-C6 alkyl, or C1-C3 alkyl. So, (C1~C 12 ) alkyl is C2-alkyl. For example, (C1-C 12 )a Alkyl is alkylene or substituted alkylene, for example, C1-C 12 alkylene, or C1~C 10 Alkylene, or C1-C6 alkylene, or C1-C3 alkyl In some cases, (C1 to C 12 ) alkyl is C2-alkylene ( For example, CH2CH2).

[0265] In certain embodiments, substituted (C1-C 12 )Alkyl is a group of 1 to 12 carbon atoms atoms, e.g., 1 to 10 carbon atoms, or 1 to 8 carbon atoms, or 1 to 6 carbon atoms elementary atom, or 1-5 carbon atoms, or 1-4 carbon atoms, or 1-3 carbon atoms A substituted alkyl group is a straight or branched chain group containing atoms. In some cases, the substituted alkyl group (C1~C 12 ) Alkyl is substituted alkyl, for example, substituted C1-C1 2 alkyl or substituted C1-C 10 Alkyl or substituted C1-C6 alkoxy In some cases, the substituted C1-C3 alkyl may be substituted or substituted C1-C3 alkyl. (C1~C 12 ) alkyl is a substituted C2-alkyl. For example, substituted ( C1~C 12 ) Alkyl is a substituted alkylene, for example, a substituted C-C 12 a alkylene or substituted C1-C 10 Alkylene or substituted C1-C6 alkyl In some cases, the alkylene may be a substituted C1-C3 alkylene. (C1~C 12 ) Alkyl is a substituted C2-alkylene.

[0266] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 and / or T 4 ) is an aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl, or substituted heterocyclyl In some cases, the tether group (e.g., T 1 , T 2, T 3 and / or T 4 ) includes aryl or substituted aryl. For example, aryl is phenyl. In some cases, the substituted aryl is a substituted phenyl. The phenyl is (C1-C 12 ) alkyl, substituted (C1-C 12 ) Alkyl, Aryl aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl cycloalkyl, heterocyclyl, and substituted heterocyclyl It may be substituted with one or more selected substituents. In some cases, the substituted The aryl is a substituted phenyl, and the substituents are a second aryl as described herein. It comprises a cleavable moiety (eg, an enzymatically cleavable moiety, eg, a glycoside).

[0267] In some cases, a tether group (e.g., T 1 , T 2 , T 3 and / or T 4 ) is hetero In some cases, the tether group (e.g., T 1 , T 2 , T 3 and / or T 4 ) is cycloalkyl or substituted cycloalkyl In some cases, a tether group (e.g., T 1 , T 2 , T 3 and / or T 4 ) includes heterocyclyl or substituted heterocyclyl. In some cases, substituted Substituents on heteroaryl, substituted cycloalkyl, or substituted heterocyclyl is cleaved with a second cleavable moiety (e.g., an enzymatically cleavable moiety) as described herein. glycosides).

[0268] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 and / or T 4 ) The tethers include ethylenediamine (EDA) moieties, e.g., EDA-containing tethers. In the embodiment, (EDA) w comprises one or more EDA moieties, e.g., w is 1 to 50; For example, 1 to 40, 1 to 30, 1 to 20, 1 to 12 or 1 to 6, e.g., 1, 2, 3, The linked ethylenediamine (EDA) moieties are integers of 1 or 2. may be substituted in any convenient position with any convenient substituent, e.g., alkyl, substituted alkyl, substituted with alkyl, acyl, substituted acyl, aryl or substituted aryl In certain embodiments, the EDA moiety may have the structure: [ka]

[0269] wherein y is an integer from 1 to 6, or 0 or 1, and each R 12 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkyl nyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted Amino, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, Aminoacyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, y is 1, 2, 3, 4, 5, or 6. In certain embodiments, y is 1, and r is 0. In certain embodiments, y is 1 and r is 1. In embodiments, y is 2 and r is 0. In certain embodiments, y is 2 and and r is 1. In certain embodiments, each R 12 is hydrogen, alkyl, substituted Independently selected from substituted alkyl, aryl and substituted aryl. In an embodiment, any two adjacent R 12 The groups may be cyclically linked, e.g., In certain embodiments, y is 1 and two aryl groups are present. Adjacent R 12 The groups are alkyl groups and are cyclically linked to form a piperazinyl ring In certain embodiments, y is 1 and the adjacent R 12 The group is hydrogen, alkyl ( e.g., methyl) and substituted alkyl (e.g., lower alkyl-OH, e.g., ethoxylated ethyl-OH or propyl-OH).

[0270] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 and / or T 4 ) is a 4-amino-piperidine (4AP) moiety (referred to herein as piperidine-4-amino, The 4AP moiety is located at one or more convenient positions. Any convenient substituents may be used, such as alkyl, substituted alkyl, polyethylene glycol, Substituted with a cole moiety, acyl, substituted acyl, aryl or substituted aryl In certain embodiments, the 4AP moiety has the structure: [ka]

[0271] Described by R 12 is hydrogen, alkyl, substituted alkyl, polyethylene glycol a cobalt moiety (e.g., polyethylene glycol or modified polyethylene glycol) ), alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy substituted alkoxy, amino, substituted amino, carboxyl, carboxyle ester, acyl, acyloxy, acylamino, aminoacyl, alkylamido, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl , substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R 12 is a polyethylene glycol moiety. In certain embodiments, R 12 is a carboxy-modified polyethylene glycol.

[0272] In certain embodiments, R 12 The structure: [ka]

[0273] The formula may be represented by: (PEG) k containing a polyethylene glycol moiety described by k is 1 to 20, for example, 1 to 18, or 1 to 16, or 1 to 14, or 1 to 1 2, or 1-10, or 1-8, or 1-6, or 1-4, or 1 or 2, For example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, In some cases, k is an integer between 16, 17, 18, 19, or 20. In some cases, k is 2. In certain embodiments, R 17 is selected from OH, COOH, or COOR, and R is Alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted hetero Aryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted In certain embodiments, R 17 is COOH .

[0274] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 and / or T 4 ) (PEG) n (PEG) n Polyethylene glycol or modified poly ethylene glycol linking units. In certain embodiments, (PEG) n The structure: [ka]

[0275] where n is 1 to 50, for example, 1 to 40, 1 to 30, 1 to 20, 1 to 12, or or 1 to 6, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, n is an integer between 14, 15, 16, 17, 18, 19, or 20. In some cases, n is 2 In some cases, n is 3. In some cases, n is 6. In some cases, where n is 12.

[0276] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 and / or T 4 ) (AA) p where AA is an amino acid residue. Any convenient amino acid may be utilized. Amino acids of interest include L- and D-amino acids, naturally occurring amino acids, and Acids, such as any of the 20 major alpha-amino acids and beta-alanine, Non-naturally occurring amino acids (e.g., amino acid analogs), e.g., non-naturally occurring amino acids amino acids or non-naturally occurring beta-amino acids. In certain embodiments, p is 1 to 50, for example, 1 to 40, 1 to 30, 1 ~20, 1-12 or 1-6, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, is an integer between 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20. In certain embodiments, p is 1. In certain embodiments, p is 2.

[0277] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 and / or T 4 ) is the formula -(CR 13 OH) h -, wherein h is 0, or n is 1 to 50, for example, 1 to 40, 1 to 30, 1 to 20, 1 to 12 or 1 to 6, for example, 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. In embodiments, h is 1. In certain embodiments, h is 2. In this state, R 13 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl alkynyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, Acylamino, aminoacyl, alkylamido, substituted alkylamido, sulfonyl , thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, hetero Aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heteroaryl In certain embodiments, substituted heterocyclyl is selected from substituted cyclohexyl, ... is R 13 is hydrogen. In certain embodiments, R 13 is alkyl or substituted Alkyl, e.g., C 1~6 Alkyl or C 1~6 substituted alkyl of, or C 1~4 Alkyl or C 1~4 substituted alkyl of, or C 1~3 Alkyl or is C 1~3 In certain embodiments, R 13 is alkenyl or substituted alkenyl, e.g., C 2~6 Alkenyl or C 2~6 is replaced by alkenyl, or C 2~4 Alkenyl or C 2~4 substituted alkenyl, or or C 2~3 Alkenyl or C 2~3 In certain embodiments, the alkenyl group is a substituted alkenyl. In the embodiment, R 13 is alkynyl or substituted alkynyl. In this state, R 13 is alkoxy or substituted alkoxy. is R 13 is amino or substituted amino. In certain embodiments, R 13 teeth carboxyl or carboxyl ester. In certain embodiments, R 13 Haa In certain embodiments, R 13 is acylamino or In certain embodiments, R 13 is an alkylamide or a substituted In certain embodiments, R 13 is sulfonyl. In certain embodiments, R 13 is thioalkoxy or substituted thioalkoxy. In certain embodiments, R 13 is aryl or substituted aryl, e.g., C 5~8 Aryl or C 5~8 substituted aryl, for example, C5 aryl or C5 substituted aryl. substituted aryl, or C6 aryl or C6 substituted aryl. In certain embodiments, R 13 is heteroaryl or substituted heteroaryl, e.g. , C 5~8 Heteroaryl or C 5~8 substituted heteroaryl, e.g., C5 heteroaryl; Heteroaryl or C5 substituted heteroaryl, or C6 heteroaryl or In certain embodiments, R 13 Cyclo Alkyl or substituted cycloalkyl, e.g., C 3~8 Cycloalkyl or C3 ~8 substituted cycloalkyl, such as C 3~6 Cycloalkyl or C 3~6 of substituted cycloalkyl, or C 3~5 Cycloalkyl or C 3~5 is replaced by In certain embodiments, R 13 is heterocyclyl or substituted C 3~8 Heterocyclyl or C 3~8 The replaced head Tetracyclyl, e.g., C 3~6 Heterocyclyl or C 3~6 Substituted heterocyclic Krill, or C 3~5 Heterocyclyl or C 3~5 Substituted heterocyclyl of be.

[0278] In certain embodiments, R 13 is hydrogen, alkyl, substituted alkyl, aryl and substituted aryl. In these embodiments, alkyl, substituted The alkyl, aryl, and substituted aryl are R 13 As mentioned above, be.

[0279] Connecting functional group, V 1 , V 2 , V 3 and V 4 With respect to which any convenient linking functional group may be First linker L 1 Linking functional groups of interest include amino groups. No, carbonyl, amide, oxycarbonyl, carboxy, sulfonyl, sulfoxide, Sulfonylamino, aminosulfonyl, thio, oxy, phospho, phosphoramidate, and and thiophosphoramidates. Well then, V 1 , V 2 , V 3 and V 4 is a covalent bond, -CO-, -NR 15 -, -NR 15 (CH2) q -, -NR 15 (C6H4)-, -CONR 15 -, -NR 15 CO-, - C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2 NR 15 -, -NR 15 and -P(O)OH-, independently selected from: and q is an integer from 1 to 6. In certain embodiments, q is an integer from 1 to 6 (e.g., 1, In certain embodiments, q is 1. In certain embodiments, q is 2, 3, 4, 5, or 6). In certain embodiments, q is 1. In an embodiment, q is 2.

[0280] In some embodiments, each R 15 is hydrogen, alkyl, substituted alkyl, alkenyl , substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted Alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl , acyloxy, acylamino, aminoacyl, alkylamido, substituted alkyla amido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted Aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl and independently selected from chloroalkyl, heterocyclyl, and substituted heterocyclyl. do.

[0281] Each R 15 The various possibilities for a particular implementation are described in more detail below. In the embodiment, R 15 is hydrogen. In certain embodiments, each R 15 is hydrogen. In certain embodiments, R 15 is alkyl or substituted alkyl, e.g., C 1~6 Alkyl or C 1~6 substituted alkyl of, or C 1~4 Alkyl or C1 ~4 substituted alkyl of, or C 1~3 Alkyl or C 1~3 Substituted Al In certain embodiments, R 15 is alkenyl or substituted alkenyl , e.g., C 2~6 Alkenyl or C 2~6 substituted alkenyl of C 2~ 4Alkenyl or C 2~4 substituted alkenyl of C 2~3 If alkenyl Kuha C 2~3 In certain embodiments, R 15 Is Alki In certain embodiments, R 15 is alkoxy or substituted alkoxy. In certain embodiments, R 15 is amino or In certain embodiments, R 15 is carboxyl or carboxy In certain embodiments, R 15 is acyl or acyloxy In certain embodiments, R 15is acylamino or aminoacyl. In certain embodiments, R 15 is an alkylamide or a substituted alkylamide. In certain embodiments, R 15 is sulfonyl. In certain embodiments, R 15 teeth In certain embodiments, R is thioalkoxy or substituted thioalkoxy. 1 5 is aryl or substituted aryl, e.g., C 5~8 Aryl or C 5~8 Placement of substituted aryl, for example, C5 aryl or C5 substituted aryl, or C In certain embodiments, R is a C6 aryl or a C6 substituted aryl. 15 is heteroaryl or substituted heteroaryl, e.g., C 5~8 Heteroaryl or or C 5~8 substituted heteroaryl, for example, C5 heteroaryl or C5 Substituted heteroaryl, or C6 heteroaryl or C6 substituted hetero In certain embodiments, R 15 is cycloalkyl or substituted cycloalkyl Chloroalkyl, e.g., C 3~8 Cycloalkyl or C 3~8 Substituted cycloalkenyl Kill, e.g., C 3~6 Cycloalkyl or C 3~6 substituted cycloalkyl of the formula: or C 3~5 Cycloalkyl or C 3~5 is a substituted cycloalkyl. In certain embodiments, R 15 is heterocyclyl or substituted heterocyclyl, e.g., Ba, C 3~8 Heterocyclyl or C3~8 substituted heterocyclyl, e.g., C3 ~6 Heterocyclyl or C 3~6 substituted heterocyclyl of C 3~5 Haitai C 3~5 is a substituted heterocyclyl of the formula:

[0282] In certain embodiments, each R 15 is hydrogen, alkyl, substituted alkyl, alkene aryl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl carboxylic esters, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and and substituted heterocyclyl. In these embodiments, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl, aryl, substituted Aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl The substituents for chloroalkyl, heterocyclyl, and substituted heterocyclyl are R 15 to This is as described above.

[0283] In certain embodiments, the tether group is an acetal group, a disulfide, a hydrazine, or In some embodiments, the tether group comprises an acetal group. In some embodiments, the tether group comprises a disulfide. In some embodiments, the tether group comprises a hydrazine. In some embodiments, the tether group comprises an ester.

[0284] As noted above, in some embodiments, L1 is -(T 1 -V 1 ) a -(T 2 -V 2 ) b -(T 3 -V 3 ) c -(T 4 -V 4 ) d a, b, c and a, b, c and d are each independently 0 or 1, and the sum of a, b, c and d is 1 to 4.

[0285] In some embodiments, the first linker L 1 In T 1 is (C1~C 12 ) alkyl and substituted (C1-C 12 ) alkyl Selected, T 2 , T 3 and T 4 is (C1~C 12 ) alkyl, substituted (C1-C 12 )a alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, Cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl Cyclyl, (EDA) w , (PEG) n , (AA) p , -(CR 13 OH) h -, 4-A Amino-piperidine (4AP), acetal group, disulfide, hydrazine, and ester are each independently selected from the V 1 , V 2 , V 3 and V 4 is a covalent bond, -CO-, -NR 15 -, -NR 15 (C H2) q -, -NR 15 (C6H4)-, -CONR 15 -, -NR 15 CO-, -C( O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR 15 -, -NR 15 and q is independently selected from SO2- and -P(O)OH-; is an integer between 1 and 6, (PEG) n teeth, [ka]

[0286] and n is an integer from 1 to 30. EDA has the following structure: [ka]

[0287] wherein y is an integer from 1 to 6, and r is 0 or 1. and 4-Amino-piperidine (4AP) is [ka]

[0288] wherein AA is an amino acid residue, p is an integer from 1 to 20, and Each R 15 and R 12 is hydrogen, alkyl, substituted alkyl, aryl and substituted and any two adjacent R 12 The groups are cyclically linked A piperazinyl ring may be formed, and R 13 is hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. The rule is selected from the following:

[0289] In certain embodiments, T 1 , T 2 , T 3 and T 4 and V 1 , V 2 , V 3 Oh BiV 4 is the following: T 1 (C1~C 12 ) alkyl and V 1 is -CO-; T 2 is 4AP and V 2 is a covalent bond; T 3 (PEG) n and V 3 is -CO-; and T 4 and V 4 does not exist is selected from.

[0290] For example, T 1 , T 2 , T 3 and T 4 and V 1 , V 2 , V 3 and V 4 is the following: T 1 is CH2CH2 and V 1 is -CO-; T 2 is 4AP and V 2 is a covalent bond; T 3 is (PEG)2 and V 3 is -CO-; and T 4 and V 4 does not exist can be selected from:

[0291] In certain embodiments, T1 , T 2 , T 3 and T 4 and V 1 , V 2 , V 3 Oh BiV 4 is the following: T 1 (C1~C 12 ) alkyl and V 1 is -CO-; T 2 is 4AP and V 2 is a covalent bond; T 3 (PEG) n and V 3 is -CONH-; and T 4 is aryl or substituted aryl, and V 4 is -CO- is selected from.

[0292] For example, T 1 , T 2 , T 3 and T 4 and V 1 , V 2 , V 3 and V 4 is the following: T 1 is CH2CH2 and V 1 is -CO-; T 2 is 4AP and V 2 is a covalent bond; T 3 is (PEG)2 and V 3 is -CONH-; and T 4 is phenyl or substituted phenyl, and V 4 is -CO- can be selected from:

[0293] In certain embodiments, the first linker L 1 has the following structure: [ka]

[0294] It is described by:

[0295] In certain embodiments, the first linker L 1 has the following structure: [ka]

[0296] It is described by:

[0297] In certain embodiments, the first linker L 1 has the following structure: [ka]

[0298] It is described by:

[0299] In certain embodiments, the left-hand side of the above linker structure is hydrazinyl-indolyl or or hydrazinyl-pyrrolo-pyridinyl coupling moiety and The right hand side of the linker structure is attached to a first cleavable moiety. When the cleavable portion of the linker structure comprises a peptide, the right hand side of the linker structure is a first cleavable In some cases, the above-mentioned amino acid sequence can be attached to a peptide containing the above-mentioned moiety. The carbonyl group on the right hand side of the linker structure is the aryl group of the peptide containing the first cleavable moiety. It can form an amide bond with an amino acid.

[0300] In some embodiments, L 2 is the formula -(L 21 ) e -(L 22 )f -(L 23 ) g -(L 24 ) h - a second linker described by L 21 , L 22 , L 23 and L 2 4 are each independently a second linker subunit, and e, f, g, and h are each independently a second linker subunit, Each of e, f, g and h is independently 0 or 1, and the sum of e, f, g and h is 0 to 4.

[0301] In certain embodiments, the sum of e, f, g, and h is 0. , a second linker L 2 does not exist. In other words, if the sum of e, f, g and h is 0, In this case, the second linker L 2 is a covalent bond. In certain embodiments, e, f, g and The sum of e, f, g and h is 1. In certain embodiments, the sum of e, f, g and h is 2. In certain embodiments, the sum of e, f, g, and h is 3. In certain embodiments, e The sum of e, f, g and h is 4. In certain embodiments, e, f, g and h are In certain embodiments, e, f, and g are each 1, and h is In certain embodiments, e and f are each 1 and g and h are In certain embodiments, e is 1 and f, g, and h are each 0. Each is 0.

[0302] In certain embodiments, L 21 is a drug (e.g., W as shown in formula (I) above). 1 ) attached to the L 22 If present, it is a drug In certain embodiments, L 23 If present, it is a drug In certain embodiments, L 24 If present, it is present in the drug It is installed.

[0303] Any convenient linker subunit can be used as the second linker L 2 May be used in Linker subunits of interest include polymeric units, e.g., polyethylene glycols. Polyol, polyethylene and polyacrylate, amino acid residue(s), carbon hydrate-based polymers or carbohydrate residues and their derivatives, polynucleotides, Alkyl groups, aryl groups, heterocyclic groups, combinations thereof, and substituted groups thereof In some embodiments, L 21 , L 22 , L 23 and L 24 Each of (if present) is polyethylene glycol, modified Polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups Linking groups including alkylene groups, substituted aryl groups, and diamines (e.g., alkylenediamines) )

[0304] In some embodiments, L 21 If present, polyethylene glycol, modified Polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups In some embodiments, L 21 Is Poly In some embodiments, L 21is a modified polyethylene glycol In some embodiments, L 21 In some embodiments, , L 21 In some embodiments, L 21 teeth In some embodiments, L 21 is diamine (e.g., a linking group comprising an alkylenediamine).

[0305] In some embodiments, L 22 If present, polyethylene glycol, modified Polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups In some embodiments, L 22 Is Poly In some embodiments, L 22 is a modified polyethylene glycol In some embodiments, L 22 In some embodiments, , L 22 In some embodiments, L 22 teeth In some embodiments, L 22 is diamine (e.g., a linking group comprising an alkylenediamine).

[0306] In some embodiments, L 23 If present, polyethylene glycol, modified Polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups In some embodiments, L 23 Is Poly In some embodiments, L 23 is a modified polyethylene glycol In some embodiments, L 23 In some embodiments, , L 23 In some embodiments, L 23 teeth In some embodiments, L 23 is diamine (e.g., a linking group comprising an alkylenediamine).

[0307] In some embodiments, L 24 If present, polyethylene glycol, modified Polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups In some embodiments, L 24 Is Poly In some embodiments, L 24 is a modified polyethylene glycol In some embodiments, L 24 In some embodiments, , L 24 In some embodiments, L 24 teeth In some embodiments, L 24 is diamine (e.g., a linking group comprising an alkylenediamine).

[0308] In some embodiments, L 2 is -(L 21 ) e -(L 22 ) f -(L 23 ) g -(L 2 4 )h a second linker comprising -(L 21 ) e -ha-(T 5 -V 5 ) e - and -(L 22 ) f -ha-(T 6 -V 6 ) f - and -(L 23 ) g -ha-(T 7 -V 7 ) g -and -(L 24 ) h -ha-(T 8 -V 8 ) h - and T 5 , T 6 , T 7 and T 8 When present, they are tether groups, V 5 , V 6 , V 7 and V 8 When present, they are linked by covalent bonds or functional groups. Yes, and e, f, g, and h are each independently 0 or 1, and the sum of e, f, g, and h is It ranges from 0 to 4.

[0309] In certain embodiments, L 21 is attached to the first detachable portion. Therefore, in certain embodiments, T 5 is attached to the first severable portion. In certain embodiments, V 5 is a drug (e.g., W as shown in formula (I) above). 1 ) In certain embodiments, L 22 If present, it is taken up by the drug Therefore, in one particular embodiment, T 6 If present, it is a drug Attached to an object or V 6 If present, it is attached to the drug In certain embodiments, L 23 If present, it is attached to the drug. Therefore, in certain embodiments, T 7 If present, it is attached to the drug Cage or V 7 If present, it is attached to the drug. In this state, L 24 If present, it is attached to the drug. In the embodiment, T 8 If present, it is attached to the drug or V 8 but If present, it is attached to a drug.

[0310] Tether group, T 5 , T 6 , T 7 and T 8 With respect to any convenient tether group, the subject phosphorus In some embodiments, the T 5 , T 6 , T 7 and T 8 Noso Each of them is (C1~C 12 ) alkyl, substituted (C1-C 12 ) Alkyl, aryl , substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, Substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (ED A) w , (PEG) n , (AA) p , -(CR 13 OH) h-, piperidine-4-amino (4AP), independently selected from acetal groups, disulfides, hydrazines, and esters. wherein w is an integer of 1 to 20, and n is an integer of 1 to 30. where p is an integer of 1 to 20 and h is an integer of 1 to 12.

[0311] In certain embodiments, a tether group (e.g., T 5 , T 6 , T 7 and / or T 8 ) (C1~C 12 ) alkyl or substituted (C1-C 12 ) alkyl. In certain embodiments, (C1 to C 12 ) Alkyl is an alkyl group having 1 to 12 carbon atoms, e.g., 1 to 10 carbon atoms, or 1 to 8 carbon atoms, or 1 to 6 carbon atoms, or 1 to Straight or branched chains containing 5 carbon atoms, or 1-4 carbon atoms, or 1-3 carbon atoms is a branched alkyl group. In some cases, (C1-C 12 ) alkyl is alkyl or substituted alkyl, e.g., C1-C 12 Alkyl, or C1-C 10 Archi It may be alkyl, C1-C6 alkyl, or C1-C3 alkyl. So, (C1~C 12 ) alkyl is C2-alkyl. For example, (C1-C 12 )a Alkyl is alkylene or substituted alkylene, for example, C1-C 12 alkylene, or C1~C 10 Alkylene, or C1-C6 alkylene, or C1-C3 alkyl In some cases, (C1 to C 12 ) alkyl is C2-alkylene ( For example, CH2CH2). In some cases, (C1-C 12 )Alkyl is C1- alkylene (e.g., CH2).

[0312] In certain embodiments, substituted (C1-C 12 )Alkyl is a group of 1 to 12 carbon atoms atoms, e.g., 1 to 10 carbon atoms, or 1 to 8 carbon atoms, or 1 to 6 carbon atoms elementary atom, or 1-5 carbon atoms, or 1-4 carbon atoms, or 1-3 carbon atoms A substituted alkyl group is a straight or branched chain group containing atoms. In some cases, the substituted alkyl group (C1~C 12 ) Alkyl is substituted alkyl, for example, substituted C1-C1 2 alkyl or substituted C1-C 10 Alkyl or substituted C1-C6 alkoxy In some cases, the alkyl group may be substituted C1-C3 alkyl. (C1~C 12 ) alkyl is a substituted C2-alkyl. For example, substituted ( C1~C 12 ) Alkyl is a substituted alkylene, for example, a substituted C-C 12 a alkylene or substituted C1-C 10 Alkylene or substituted C1-C6 alkyl In some cases, the alkylene may be a substituted C1-C3 alkylene. (C1~C 12 ) alkyl is a substituted C2-alkylene. , substituted (C1~C 12 ) Alkyl is a substituted C1-alkylene.

[0313] In certain embodiments, a tether group (e.g., T 5 , T 6 , T7 and / or T 8 ) is an aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl, or substituted heterocyclyl In some cases, the tether group comprises an aryl or a substituted aryl. For example, aryl can be phenyl or substituted phenyl. is a tether group (e.g., T 5 , T 6 , T 7 and / or T 8 ) is heteroaryl or In some cases, the tether group (e.g., T 5 , T 6 , T 7 and / or T 8 ) includes cycloalkyl or substituted cycloalkyl. In this case, the tether group (e.g., T 5 , T 6 , T 7 and / or T 8 ) is heterocycly aryl or substituted heterocyclyl.

[0314] In certain embodiments, a tether group (e.g., T 5 , T 6 , T 7 and / or T 8 ) is an ethylenediamine (EDA) moiety, e.g., an EDA moiety as described above, e.g., the structure : [ka]

[0315] and comprising an EDA moiety described by y is an integer from 1 to 6, or 0 or 1, and each R12 is hydrogen, alkyl alkyl, alkenyl, substituted alkenyl, alkynyl, substituted Alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxy Cyl, carboxyl ester, acyl, acyloxy, acylamino, aminoacyl, acyl alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thia Alkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted cycloalkyl In certain embodiments, y is independently selected from 1, 2, 3, In certain embodiments, y is 1 and r is 0. In certain embodiments, y is 1 and r is 1. In certain embodiments, y is 1 and r is 1. is 2 and r is 0. In certain embodiments, y is 2 and r is 1. In certain embodiments, each R 12 is hydrogen, alkyl, substituted alkyl, In certain embodiments, E is independently selected from aryl and substituted aryl. Any two adjacent R in DA 12 The groups may be cyclically linked to form, for example, a piperazinyl ring. In certain embodiments, y is 1 and two adjacent R 12 The groups are alkyl groups and are cyclically linked to form a piperazinyl ring. In the form, y is 1 and adjacent R 12 The group can be hydrogen, alkyl (e.g., methyl), and substituted alkyl (e.g., lower alkyl-OH, e.g., ethyl-OH or propyl-OH).

[0316] In certain embodiments, a tether group (e.g., T 5 , T 6 , T 7 and / or T 8 ) is a 4-amino-piperidine (4AP) moiety as described above, for example, the structure: [ka]

[0317] and comprising a 4AP moiety described by R 12 is a hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety (e.g., For example, polyethylene glycol or modified polyethylene glycol), alkenyl, Substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkenyl Alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, Acyloxy, acylamino, aminoacyl, alkylamido, substituted alkylamino alkyl, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted alkyl aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cyclo In particular, the alkyl group is selected from alkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R 12 is a polyethylene glycol moiety. is R 12 is a carboxy-modified polyethylene glycol.

[0318] In certain embodiments, R 12 The structure: [ka]

[0319] The formula may be represented by: (PEG) k containing a polyethylene glycol moiety described by k is 1 to 20, for example, 1 to 18, or 1 to 16, or 1 to 14, or 1 to 1 2, or 1-10, or 1-8, or 1-6, or 1-4, or 1 or 2, For example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, In some cases, k is an integer between 16, 17, 18, 19, or 20. In some cases, k is 2. In certain embodiments, R 17 is selected from OH, COOH, or COOR, and R is Alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted hetero Aryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted In certain embodiments, R 17 is COOH .

[0320] In certain embodiments, a tether group (e.g., T 5 , T 6 , T 7 and / or T 8 ) is a polyethylene glycol moiety (PEG) as described above. n , for example, the structure: [ka]

[0321] (PEG) n Including parts, n is 1 to 50, for example, 1 to 40, 1 to 30, 1 to 20, 1 to 12 or 1 to 6, e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 , 17, 18, 19, or 20. In some cases, n is 2. In some cases, n is 3. In some cases, n is 6. In some cases, n is 12. do.

[0322] In certain embodiments, a tether group (e.g., T 5 , T 6 , T 7 and / or T 8 ) (AA) p where AA is an amino acid residue. Any convenient amino acid may be utilized. Amino acids of interest include L- and D-amino acids, naturally occurring amino acids, and Acids, such as any of the 20 major alpha-amino acids and beta-alanine, Non-naturally occurring amino acids (e.g., amino acid analogs), e.g., non-naturally occurring amino acids amino acids or non-naturally occurring beta-amino acids. In certain embodiments, p is 1 to 50, for example, 1 to 40, 1 to 30, 1 ~20, 1-12 or 1-6, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, is an integer between 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20. In certain embodiments, p is 1. In certain embodiments, p is 2.

[0323] In certain embodiments, a tether group (e.g., T 5 , T 6 , T 7 and / or T 8 ) is the formula -(CR 13 OH) h -, wherein h is 0, or n is 1 to 50, for example, 1 to 40, 1 to 30, 1 to 20, 1 to 12 or 1 to 6, for example, 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. In embodiments, h is 1. In certain embodiments, h is 2. In this state, R 13 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl alkynyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, Acylamino, aminoacyl, alkylamido, substituted alkylamido, sulfonyl , thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, hetero Aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heteroaryl In certain embodiments, substituted heterocyclyl is selected from substituted cyclohexyl, ... is R 13 is hydrogen. In certain embodiments, R 13 is alkyl or substituted Alkyl, e.g., C 1~6 Alkyl or C 1~6 substituted alkyl of, or C 1~4 Alkyl or C 1~4 substituted alkyl of, or C 1~3 Alkyl or is C 1~3 In certain embodiments, R 13 is alkenyl or substituted alkenyl, e.g., C 2~6 Alkenyl or C 2~6 is replaced by alkenyl, or C 2~4 Alkenyl or C 2~4 substituted alkenyl, or or C2~3 Alkenyl or C 2~3 In certain embodiments, the alkenyl group is a substituted alkenyl. In the embodiment, R 13 is alkynyl or substituted alkynyl. In this state, R 13 is alkoxy or substituted alkoxy. is R 13 is amino or substituted amino. In certain embodiments, R 13 teeth carboxyl or carboxyl ester. In certain embodiments, R 13 Haa In certain embodiments, R 13 is acylamino or In certain embodiments, R 13 is an alkylamide or a substituted In certain embodiments, R 13 is sulfonyl. In certain embodiments, R 13 is thioalkoxy or substituted thioalkoxy. In certain embodiments, R 13 is aryl or substituted aryl, e.g., C 5~8 Aryl or C 5~8 substituted aryl, for example, C5 aryl or C5 substituted aryl. substituted aryl, or C6 aryl or C6 substituted aryl. In certain embodiments, R 13 is heteroaryl or substituted heteroaryl, e.g. , C 5~8 Heteroaryl or C 5~8 substituted heteroaryl, e.g., C5 heteroaryl; Heteroaryl or C5 substituted heteroaryl, or C6 heteroaryl or In certain embodiments, R 13 Cyclo Alkyl or substituted cycloalkyl, e.g., C 3~8 Cycloalkyl or C3 ~8 substituted cycloalkyl, such as C 3~6 Cycloalkyl or C 3~6 of substituted cycloalkyl, or C 3~5 Cycloalkyl or C 3~5 is replaced by In certain embodiments, R 13 is heterocyclyl or substituted C 3~8 Heterocyclyl or C 3~8 The replaced head Tetracyclyl, e.g., C 3~6 Heterocyclyl or C 3~6 Substituted heterocyclic Krill, or C 3~5 Heterocyclyl or C 3~5 Substituted heterocyclyl of be.

[0324] In certain embodiments, R 13 is hydrogen, alkyl, substituted alkyl, aryl and substituted aryl. In these embodiments, alkyl, substituted The alkyl, aryl, and substituted aryl are R 13 As mentioned above, be.

[0325] Connecting functional group, V 5 , V 6 , V 7 and V 8 With respect to which any convenient linking functional group may be Second linker L 2 Linking functional groups of interest include amino groups. No, carbonyl, amide, oxycarbonyl, carboxy, sulfonyl, sulfoxide, Sulfonylamino, aminosulfonyl, thio, oxy, phospho, phosphoramidate, and and thiophosphoramidates. Well then, V 5 , V 6 , V 7 and V 8 is a covalent bond, -CO-, -NR 15 -, -NR 15 (CH2) q -, -NR 15 (C6H4)-, -CONR 15 -, -NR 15 CO-, - C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2 NR 15 -, -NR 15 and -P(O)OH-, independently selected from: and q is an integer from 1 to 6. In certain embodiments, q is an integer from 1 to 6 (e.g., 1, In certain embodiments, q is 1. In certain embodiments, q is 2, 3, 4, 5, or 6). In certain embodiments, q is 1. In an embodiment, q is 2.

[0326] In some embodiments, each R 15 is hydrogen, alkyl, substituted alkyl, alkenyl , substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted Alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl , acyloxy, acylamino, aminoacyl, alkylamido, substituted alkyla amido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted Aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl and independently selected from chloroalkyl, heterocyclyl, and substituted heterocyclyl. do.

[0327] Each R 15 The various possibilities for a particular implementation are described in more detail below. In the embodiment, R 15 is hydrogen. In certain embodiments, each R 15 is hydrogen. In certain embodiments, R 15 is alkyl or substituted alkyl, e.g., C 1~6 Alkyl or C 1~6 substituted alkyl of, or C 1~4 Alkyl or C1 ~4 substituted alkyl of, or C 1~3 Alkyl or C 1~3 Substituted Al In certain embodiments, R 15 is alkenyl or substituted alkenyl , e.g., C 2~6 Alkenyl or C 2~6 substituted alkenyl of C 2~ 4Alkenyl or C 2~4 substituted alkenyl of C 2~3 If alkenyl Kuha C 2~3 In certain embodiments, R 15 Is Alki In certain embodiments, R 15 is alkoxy or substituted alkoxy. In certain embodiments, R 15 is amino or In certain embodiments, R 15 is carboxyl or carboxy In certain embodiments, R 15 is acyl or acyloxy In certain embodiments, R 15 is acylamino or aminoacyl. In certain embodiments, R 15 is an alkylamide or a substituted alkylamide. In certain embodiments, R 15 is sulfonyl. In certain embodiments, R 15 teeth In certain embodiments, R is thioalkoxy or substituted thioalkoxy. 1 5 is aryl or substituted aryl, e.g., C 5~8 Aryl or C 5~8 Placement of substituted aryl, for example, C5 aryl or C5 substituted aryl, or C In certain embodiments, R is a C6 aryl or a C6 substituted aryl. 15 is heteroaryl or substituted heteroaryl, e.g., C 5~8 Heteroaryl or or C 5~8 substituted heteroaryl, for example, C5 heteroaryl or C5 Substituted heteroaryl, or C6 heteroaryl or C6 substituted hetero In certain embodiments, R 15 is cycloalkyl or substituted cycloalkyl Chloroalkyl, e.g., C 3~8 Cycloalkyl or C 3~8 Substituted cycloalkenyl Kill, e.g., C 3~6 Cycloalkyl or C 3~6 substituted cycloalkyl of the formula: or C 3~5 Cycloalkyl or C 3~5 is a substituted cycloalkyl. In certain embodiments, R 15 is heterocyclyl or substituted heterocyclyl, e.g., Ba, C 3~8 Heterocyclyl or C 3~8 substituted heterocyclyl, e.g., C3 ~6 Heterocyclyl or C 3~6 substituted heterocyclyl of C 3~5 Haitai C 3~5 is a substituted heterocyclyl of the formula:

[0328] In certain embodiments, each R 15 is hydrogen, alkyl, substituted alkyl, alkene aryl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl carboxylic esters, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and and substituted heterocyclyl. In these embodiments, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl, aryl, substituted Aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl The substituents for chloroalkyl, heterocyclyl, and substituted heterocyclyl are R 15 to This is as described above.

[0329] In certain embodiments, the tether group is an acetal group, a disulfide, a hydrazine, or In some embodiments, the tether group comprises an acetal group. In some embodiments, the tether group comprises a disulfide. In some embodiments, the tether group comprises a hydrazine. In some embodiments, the tether group comprises an ester.

[0330] As noted above, in some embodiments, L 2 is -(T 5 -V 5 ) e -(T 6 -V 6 ) f -(T 7 -V 7 ) g -(T 8 -V 8 ) h -, and a second linker comprising e, f, g and e, f, g and h each independently represent 0 or 1, and the sum of e, f, g and h is 0 to 4.

[0331] In some embodiments, the second linker L 2 In T 5 , T 6 , T 7 and T 8 is (C1~C 12 ) alkyl, substituted (C1-C1 2) Alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl cycloalkyl, substituted cycloalkyl, heterocyclyl, or substituted Heterocyclyl, (EDA) w , (PEG) n , (AA) p , -(CR 13 OH) h -, 4-amino-piperidine (4AP), acetal groups, disulfides, hydrazines, and esters, and V 5 , V 6 , V 7 and V 8 is a covalent bond, -CO-, -NR 15 -, -NR 15 (C H2) q -, -NR 15 (C6H4)-, -CONR15 -, -NR 15 CO-, -C( O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR 15 -, -NR 15 and q is independently selected from SO2- and -P(O)OH-; is an integer between 1 and 6, (PEG) n teeth, [ka]

[0332] and n is an integer from 1 to 30. EDA has the following structure: [ka]

[0333] wherein y is an integer from 1 to 6, and r is 0 or 1. and 4-Amino-piperidine (4AP) is [ka]

[0334] wherein AA is an amino acid residue and p is an integer of 1 to 20; Each R 13 is hydrogen, alkyl, substituted alkyl, aryl, and substituted arylene. are selected independently from the rule, and Each R 15 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl , aryl, substituted aryl, heteroaryl, substituted heteroaryl, cyclo Alkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl are selected independently from the

[0335] In certain embodiments, T 5 , T 6 , T 7 and T 8 and V 5 , V 6 , V 7 Oh BiV 8 is the following: T 5 is a covalent bond, and V 5 is -OC(O)-; T 6 and V 6 does not exist; T 7 and V 7 does not exist; and T 8 and V 8 does not exist is selected from.

[0336] In certain embodiments, T 5 , T 6 , T 7 and T 8 and V 5 , V 6 , V 7 Oh BiV 8 does not exist (i.e., the sum of e, f, g, and h is 0).

[0337] In certain embodiments, the left-hand side of the above linker structure is attached to a first cleavable moiety. The right hand side of the linker structure is attached to the drug.

[0338] Any chemical entities, linkers and coupling moieties shown in the structures above may be used in conjunction with the subject matter. The compounds may be adapted for use in combinations and conjugates.

[0339] Hydrazinyl-indolyl and hydrazinyl-pyrrolo-pyridinyl compounds and Additional disclosure on methods for making conjugates can be found in U.S. Pat. No. 9,310,374. and U.S. Pat. No. 9,493,413, the respective disclosures of which are incorporated herein by reference. is incorporated herein by reference.

[0340] Compounds useful for preparing conjugates The present disclosure provides hydrazinyl-isocyanates useful for preparing the conjugates described herein. In certain embodiments, the compounds provided herein are hydrazinyl-pyrrolo-pyridinyl compounds. In this embodiment, the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl compound is It may also be a coupling moiety useful for conjugating antibodies and drugs. For example, hydra the indolyl- or hydrazinyl-pyrrolo-pyridinyl compound is conjugated to the antibody, The antibody may also be bound to a drug, thereby indirectly binding the antibody and the drug.

[0341] In certain embodiments, the compound has formula (II): [ka]

[0342] is a compound of the formula Z is CR 4 or N, R 2 and R 3 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, a amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy , acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl aryl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, hetero aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; is R 2 and R 3 may be cyclically linked to form a 5- or 6-membered heterocyclyl. Ku, Each R 4 is hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, Amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy Acylamino, aminoacyl, alkylamido, substituted alkylamido, sulfo aryl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, hydroxyl Heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl , heterocyclyl, and substituted heterocyclyl; Each R 5 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl , alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl , substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclo and substituted heterocyclyl; Each R 6 is an alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkyn ... alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; k is an integer from 1 to 10, R 7 is hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, a amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy , acylamino, aminoacyl, alkylamido, substituted alkylamido, aryl , substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, Selected from substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl And, L 1 is the first linker, L 2 is a second linker, and W 1 is a drug, L 1 , R 6 or R 7 One of the fragments comprises a second cleavable portion.

[0343] In some instances, k is 2 and the compound has the formula (IIa): [ka]

[0344] is a compound of the formula R 6’ or R 6’’ one of which contains a second cleavable moiety, and R 6’ and R 6’’ The other of the groups is alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heteroaryl and substituted heterocyclyl.

[0345] For example, the compound has the formula (IIb): [ka]

[0346] It may also be a compound of the formula:

[0347] In some instances, the compound has formula (IIc): [ka]

[0348] It may also be a compound of the formula:

[0349] In certain embodiments, k is 2 and the compound has formula (IId): [ka]

[0350] is a compound of the formula R 6’ and R 6’’ is an alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, hetero Aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heteroaryl independently selected from tetracyclyl, tetracyclyl, and substituted heterocyclyl; and R 7 comprises a second cleavable moiety.

[0351] For example, the compound has the formula (IIe): [ka]

[0352] It may also be a compound of the formula:

[0353] In certain embodiments, k is 2 and the compound has the formula (IIf): [ka]

[0354] is a compound of the formula R 6’ and R 6’’ is an alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, hetero Aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heteroaryl independently selected from tetracyclyl, tetracyclyl, and substituted heterocyclyl; and R 8 comprises a second cleavable moiety.

[0355] For example, the compound has the formula (IIg): [ka]

[0356] It may also be a compound of the formula:

[0357] Substituents for the conjugate of formula (II) are described herein. Reference to formula (IIa), (IIb), (IIc), (IId), (IIe), (II It is intended to encompass (Ig) and (IIg).

[0358] With respect to the compounds of formula (II), the substituents Z, R 2 , R3 , R 4 , R 5 , L 1 , L 2 , and BiW 1 is as described above for the conjugate of formula (I). Similarly, ) the first linker L 1 and a second linker L 2 Regarding T 1 , T 2 , T 3 , T 4 , V 1 , V 2 , V 3 , and V 4 , and T 5 , T 6 , T 7 , T 8 , V 5 , V 6 , V 7 Oh BiV 8 The substituents of are as described above for the conjugates of formula (I).

[0359] In certain embodiments, the compound has the following structure: [ka]

[0360] is a compound of

[0361] For example, the compound may have the following structure: [ka]

[0362] It can have:

[0363] In certain embodiments, the compound has the following structure: [ka]

[0364] is a compound of

[0365] For example, the compound may have the following structure: [ka]

[0366] It can have:

[0367] In certain embodiments, the compound has the following structure: [ka]

[0368] is a compound of

[0369] For example, the compound may have the following structure: [ka]

[0370] It can have:

[0371] In certain embodiments, the compound has the following structure: [ka]

[0372] is a compound of

[0373] For example, the compound may have the following structure: [ka]

[0374] It can have:

[0375] In certain embodiments, the compound has the following structure: [ka]

[0376] is a compound of

[0377] For example, the compound may have the following structure: [ka]

[0378] It can have:

[0379] antibody As noted above, the subject conjugates may contain W as a substituent. 2 It may contain antibodies, The antibody has been modified to include a 2-formylglycine (FGly) residue. When used in this document, amino acids are indicated by their standard names, their standard three-letter abbreviations, and / or their standard one-letter abbreviations, e.g., alanine or Ala or A ;Cysteine ​​or Cys or C;Aspartic acid or Asp or D;Glutamine Acid or Glu or E; Phenylalanine or Phe or F; Glycine or GI y or G; histidine or His or H; isoleucine or Ile or I; sine or Lys or K; leucine or Leu or L; methionine or Met or or M; asparagine or Asn or N; proline or Pro or P; glutamine Gln or Gln or Q; Arginine or Arg or R; Serine or Ser or S; threonine or Thr or T; valine or Val or V; tryptophan or or Trp or W, and tyrosine or Tyr or Y .

[0380] In certain embodiments, the amino acid sequence of the antibody is selected from those expressed in vivo (e.g., in cells). at the time of translation of aldehyde-tagged proteins) or in vitro (e.g., in cellular In a cellular system, aldehyde-tagged proteins are conjugated with formylglycine generating enzyme (FGE). 2-formylglycine (FGly) is produced by the action of FGE either by contact with the sulfur containing serine or cysteine ​​residues that can be converted (oxidized) to cysteine ​​residues. Such a sulfatase motif may also be modified to include In this specification, this may be referred to as an FGE modification site.

[0381] sulfatase motif The minimal sulfatase motif for the aldehyde tag is usually 5 or 6 amino acid residues long The sulfatase fragments provided in Ig polypeptides are usually six amino acid residues or less in length. The ze motif is at least 5 or 6 amino acid residues, e.g., 5-16, 6-16, 5~15, 6~15, 5~14, 6~14, 5~13, 6~13, 5~12, 6~12, 5-11, 6-11, 5-10, 6-10, 5-9, 6-9, 5-8, or 6-8 mesh The length of the amino acid residues can be 16, 15, 14, 13, 12, 11, Sulfatase motifs of less than 10, 9, 8, 7, or 6 amino acid residues in length are defined. can be.

[0382] In certain embodiments, the polypeptide of interest may have a native amino acid sequence. one or more amino acid residues, e.g., two or more, or three or more, or 4 or more, or 5 or more many, or six or more, or seven or more, or eight or more, or 9 or more, or 10 or more or 11 or more, or 12 or more, or or 13 or more, or 14 or more, or 15 or or more, or 16 or more, or 17 or more many, or 18 or more, or 19 or more, and is a polynucleotide consisting of 20 or more amino acid residues inserted, deleted, or substituted (replaced) Examples include peptides that provide the sequence of a sulfatase motif. In this embodiment, the polypeptide has an amino acid sequence that is different from the native amino acid sequence of the polypeptide. 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, and 9 amino acid sequences , 8, 7, 6, 5, 4, 3 or fewer than 2 amino acid residues (insertion, addition, deletion, and and / or substitutions / replacements) of the native amino acid sequence of a polypeptide (e.g., an antibody). If the amino acid sequence contains one or more residues of the desired sulfatase motif, modification of the residues is The total number of mutations may be, for example, 100 or more, depending on the native sequence to provide the desired sulfatase motif sequence. Site-specific modification (insertion, addition, deletion, substitution / alteration) of amino acid residues adjacent to the target amino acid residue In certain embodiments, the native nucleotide sequence of the target antibody can be reduced by The degree of modification of the amino acid sequence may be insertion, deletion, substitution (replacement), or addition (e.g., The target antigen is minimized to minimize the number of amino acid residues that are transferred (to the N- or C-terminus). Minimizing the degree of amino acid sequence modifications of the antibody is important because such modifications may adversely affect antibody function and / or structure. This may minimize the influence that the organization may have on the organization.

[0383] Aldehyde tags of particular interest should contain at least the minimal sulfatase motif ("co- These contain a longer amino acid sequence (also called the "consensus sulfatase motif"). Dehyde tags are contemplated and encompassed by the present disclosure and may be used in the compositions and methods of the present disclosure. It should be noted that it is readily apparent that the aldehyde tag may have the following structure: Therefore, it can contain a minimal sulfatase motif of 5 or 6 residues, or more long and flanked on the N- and / or C-terminal sides of the motif by additional amino acid residues The smallest sulfatase motif that can be obtained is, for example, 5 or 6. In addition to amino acid residues 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, and 16 , longer amino acids of more than 17, 18, 19, 20 or more amino acid residues An aldehyde tag of the acid sequence is envisaged.

[0384] The aldehyde tag can be at or near the C-terminus of the Ig heavy chain, e.g., The aldehyde tag is located at the C-terminus of the native wild-type Ig heavy chain at positions 1, 2, 3, 4, 5, 6, and 7. The aldehyde tag can be within 1, 8, 9, or 10 amino acids of the Ig heavy The aldehyde tag can be located in the CH1 domain of the Ig heavy chain. The aldehyde tag can be located in the CH3 domain of the Ig heavy chain. The aldehyde tag can be present in the Ig light chain constant region, e.g., the kappa light chain constant region. The nucleotide sequence may be present in the lambda light chain constant region or the lambda light chain constant region.

[0385] In certain embodiments, the sulfatase motif used has the formula: X 1 Z 10 X 2 Z 20 X 3 Z 30 (I') wherein: Z 10 is cysteine ​​or serine (also represented by (C / S)), Z 20 is either a proline or alanine residue (also represented by (P / A) (can be), Z 30 is a basic amino acid (e.g., arginine (R) and not lysine (K) or hyaluronan) stidine (H), which may be, for example, lysine), or an aliphatic amino acid (alanine ( A), glycine (G), leucine (L), valine (V), isoleucine (I), or proline (P), e.g., A, G, L, V, or I; X 1 may or may not be present, and if present, may be any amino acid, e.g., an aliphatic amino acid. amino acids, sulfur-containing amino acids, or polar uncharged amino acids (i.e., aromatic amino acids or or charged amino acids), e.g., L, M, V, S, or T, e.g., L, M, S, or can be V, provided that the sulfatase motif is at the N-terminus of the target polypeptide. If so, X 1 exists and X 2 and X 3 can independently be any amino acid, but are typically aliphatic amino acids. acids, polar uncharged amino acids, or sulfur-containing amino acids (i.e., aromatic amino acids or Other than charged amino acids), e.g., S, T, A, V, G or C, e.g., S, T, A, V Or G.

[0386] The amino acid sequence of the antibody heavy and / or light chains may be represented by the formula X 1 Z 10 X 2 Z 20 X 3 Z 30 of can be modified to provide a sequence of at least 5 amino acids, wherein Z 10 is cysteine ​​or serine, Z 20 is a proline or alanine residue, Z 30 is an aliphatic amino acid or a basic amino acid, X 1 is present or absent, and if present, is any amino acid, except for heterologous sequences. If the sulfatase motif is at the N-terminus of the polypeptide, X 1 exists, X 2 and X 3 are each independently any amino acid.

[0387] The sulfatase motif is generally selected from the FGE of choice, e.g., aldehyde tagging. FGE or cell-free in vitro methods in which the polypeptide is expressed in a host cell This can be converted by FGE in which the protein is contacted with an aldehyde-tagged polypeptide. It is selected to be.

[0388] For example, the FGE is a eukaryotic FGE (e.g., a mammalian FGE, including a human FGE). where the sulfatase motif has the formula: X 1 CX 2 PX 3 Z 30 (I'') wherein: X 1 may or may not be present, and if present, may be any amino acid, e.g., a fatty acid. aromatic amino acids, sulfur-containing amino acids, or polar uncharged amino acids (i.e., aromatic amino acids) amino acids or charged amino acids), e.g., L, M, S, or V, provided that , if the sulfatase motif is at the N-terminus of the target polypeptide, X 1 exists, X 2 and X 3 are independently any amino acid, e.g., aliphatic amino acids, sulfur-containing amino acids, acids, or polar uncharged amino acids (i.e., aromatic amino acids or charged amino acids) outside), e.g., S, T, A, V, G, or C, e.g., S, T, A, V, or G can be, and Z 30 is a basic amino acid (e.g., arginine (R) and not lysine (K) or hyaluronan) stidine (H), which may be, for example, lysine), or an aliphatic amino acid (alanine ( A), glycine (G), leucine (L), valine (V), isoleucine (I), or Proline (P), e.g., A, G, L, V, or I.

[0389] A specific example of a sulfatase motif is LCTPSR (SEQ ID NO: / ), MCTPSR(SEQ ID NO: / / ), VCTPSR(SEQ ID NO : / / ), LCSPSR(SEQ ID NO: / / ), LCAPSR(SEQ ID NO: / / ), LCVPSR(SEQ ID NO: / / ), LCGPSR(SEQ I D NO: / / ), ICTPAR(SEQ ID NO: / / ), LCTPSK(SEQ ID NO: / / ), MCTPSK(SEQ ID NO: / / ), VCTPSK(S EQ ID NO: / / ), LCSPSK(SEQ ID NO: / / ), LCAPSK (SEQ ID NO: / / ), LCVPSK(SEQ ID NO: / / ), LCGP SK(SEQ ID NO: / / ), LCTPSA(SEQ ID NO: / / ), IC TPAA(SEQ ID NO: / / ), MCTPSA(SEQ ID NO: / / ), VCTPSA(SEQ ID NO: / / ), LCSPSA(SEQ ID NO: / / ) ), LCAPSA(SEQ ID NO: / / ), LCVPSA(SEQ ID NO: / / ), and LCGPSA(SEQ ID NO: / / ).

[0390] FGly-containing sequence The action of FGE on the engineered antibody heavy and / or light chains results in the formation of a sulfatase motif The serine or cysteine ​​in the ATP is modified to FGly. The tase motif has the formula: X 1 (FGly)X 2 Z 20 X 3 Z 30 (I''') wherein: FGly is a formylglycine residue, Z 20 is either a proline or alanine residue (also represented by (P / A) (can be), Z 30 is a basic amino acid (e.g., arginine (R) and not lysine (K) or hyaluronan) stigmine (H), usually lysine), or an aliphatic amino acid (alanine (A) , glycine (G), leucine (L), valine (V), isoleucine (I), or proline phosphorus (P), e.g., A, G, L, V, or I; X 1 may or may not be present, and if present, may be any amino acid, e.g., a fatty acid. aromatic amino acids, sulfur-containing amino acids, or polar uncharged amino acids (i.e., aromatic amino acids) other than amino acids or charged amino acids), e.g., L, M, V, S, or T, e.g., L, M, or or V, provided that the sulfatase motif is at the N-terminus of the target polypeptide. If yes, X 1 exists and X 2 and X 3 are independently any amino acid, e.g., aliphatic amino acids, sulfur-containing amino acids, acids, or polar uncharged amino acids (i.e., aromatic amino acids or charged amino acids) Other), e.g., S, T, A, V, G or C, e.g., S, T, A, V or G This can be done.

[0391] As described above, modified polypeptides containing FGly residues are FGly'-containing polypeptides. A drug to generate the sulfatase motif (e.g., hydrazinyl-isothiazolinone, as described above) and drugs containing an indolyl or hydrazinyl-pyrrolo-pyridinyl coupling moiety. The FGly may be conjugated to a drug (e.g., maytansinoid) by reaction of the FGly. As used herein, the term FGly' refers to a drug, e.g., maytansine or or modified amino acids in the sulfatase motif coupled to auristatin Thus, the FGly'-containing sulfatase motif has the formula: X 1 (FGly')X 2 Z 20 X 3 Z 30 (II) wherein: FGly' is a modified amino acid residue of formula (I), Z 20 is either a proline or alanine residue (also represented by (P / A) (can be), Z 30 is a basic amino acid (e.g., arginine (R) and not lysine (K) or hyaluronan) stigmine (H), usually lysine), or an aliphatic amino acid (alanine (A) , glycine (G), leucine (L), valine (V), isoleucine (I), or proline phosphorus (P), e.g., A, G, L, V, or I; X 1 may or may not be present, and if present, may be any amino acid, e.g., a fatty acid. aromatic amino acids, sulfur-containing amino acids, or polar uncharged amino acids (i.e., aromatic amino acids) other than amino acids or charged amino acids), e.g., L, M, V, S, or T, e.g., L, M, or or V, provided that the sulfatase motif is at the N-terminus of the target polypeptide. If yes, X 1 exists and X 2 and X 3 are independently any amino acid, e.g., aliphatic amino acids, sulfur-containing amino acids, acids, or polar uncharged amino acids (i.e., aromatic amino acids or charged amino acids) Other), e.g., S, T, A, V, G or C, e.g., S, T, A, V or G This can be done.

[0392] Modification site As mentioned above, the amino acid sequence of the antibody is determined by the aldehyde dehydrogenase (DA) in vivo (e.g., in cells). at the time of translation of the tag-containing protein) or in vitro (e.g., in a cell-free system) by contacting the aldehyde-tagged protein with FGE) Serine or cysteine ​​that can be converted (oxidized) to a FGly residue by the action of E The conjugates of the present disclosure are modified to include a sulfatase motif containing residues. The antibody used to generate the antibody should contain at least an Ig constant region, e.g., an Ig heavy chain constant region. regions (e.g., at least the CH1 domain; at least the CH1 and CH2 domains; C H1, CH2, and CH3 domains; or CH1, CH2, CH3, and CH4 domain), or an Ig light chain constant region. Such Ig polypeptides are referred to herein as These are referred to in the literature as "target Ig polypeptides" or "target antibodies."

[0393] The site in the antibody into which the sulfatase motif is introduced can be any convenient site. As noted above, in some cases, the native amino acids of the target polypeptide can be The extent of sequence modification may include insertions, deletions, substitutions (replacements), and / or additions (e.g., N The target antibody is minimized to minimize the number of amino acid residues (e.g., to the C-terminus). Minimizing the extent of amino acid sequence modifications is important because such modifications may have an adverse effect on antibody function and / or structure. This may minimize the influence they may have on the

[0394] The antibody heavy chain constant region may be an Ig constant region of any heavy chain isotype, a non-naturally occurring Ig The Ig constant region may comprise a heavy chain constant region (including a consensus Ig heavy chain constant region). The region can be modified to include an aldehyde tag, which is an Ig constant. The Ig constant region is located in or adjacent to the solvent accessible loop region. To provide the amino acid sequence of the sulfatase motif as described above, , 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 amino acids, Or it can be modified by the insertion and / or substitution of more than 16 amino acids.

[0395] In some cases, the aldehyde-tagged antibody comprises an aldehyde-tagged Ig heavy chain constant region ( For example, at least a CH1 domain; at least a CH1 and a CH2 domain; CH1, CH2, and CH3 domains; or CH1, CH2, CH3, and CH4 domains The aldehyde-tagged Ig heavy chain constant region is modified by FGE to form the FGly modification. at least one sulfatase motif capable of producing a decorated Ig polypeptide. Modified to include IgA, IgM, IgD, IgE, IgG1, IgG2, IgG 3, or the heavy chain constant region sequence of an IgG4 isotype heavy chain or any allotype thereof Variants, e.g., human or mouse heavy chain constant region sequences, hybrids heavy chain constant region, synthetic heavy chain constant region, or consensus heavy chain constant region sequence, etc. Allotypic variants of Ig heavy chains are known in the art. See, for example, Jefferis and Lefranc (2009) MAbs 1:4 .

[0396] In some cases, the aldehyde-tagged antibody comprises an aldehyde-tagged Ig light chain constant region. The aldehyde-tagged Ig light chain constant region is modified by FGE to produce FGly-modified antibodies. and modified to contain at least one sulfatase motif capable of generating Also, kappa light chain and lambda light chain constant region sequences, e.g., human kappa or lambda light chain constant region sequences, constant region, hybrid light chain constant region, synthetic light chain constant region, or consensus light chain constant region Exemplary constant regions include human gamma 1 and gamma 2 constant regions. With the exception of the sulfatase motif, the modified constant regions are the wild-type The amino acid sequence may be at least 70% identical to the wild-type amino acid sequence. an identical (e.g., at least 80%, at least 90%, or at least 95% identical) It may have an amino acid sequence.

[0397] In some embodiments, the sulfatase motif is located at a site other than the C-terminus of the Ig polypeptide heavy chain. As described above, the isolated aldehyde tagged The antibody comprises a heavy chain constant region modified to contain a sulfatase motif as described above. The sulfatase motif can be found in a surface accessible loop of the antibody heavy chain constant region. Located in or adjacent to the region.

[0398] The sulfatase motif is a sequence of these amino acids at the site of such modification in the Ig heavy chain. It can be provided within or adjacent to one or more of the Ig heavy chain polypeptides. The peptides are located adjacent to these modification sites and at the N-terminus and / or adjacent to the To provide a sulfatase motif at the C-terminus, one or more of these amino acid sequences may be used. Modifications in number (e.g., modifications include insertions, deletions, and / or deletions of one or more amino acid residues) Alternatively or additionally, the Ig heavy chain polypeptide may be to provide a sulfatase motif between any two residues of the Ig heavy chain modification site. Modifications in one or more of these amino acid sequences (e.g., modifications include one or more amino acids In some embodiments, the amino acid residues may be inserted, deleted, and / or substituted. In one embodiment, the Ig heavy chain polypeptide may be modified to include two motifs, The motifs may be adjacent to each other or may be 1, 2, 3, 4 or more. (e.g., about 1 to about 25, about 25 to about 50, or about 50 to about 100, or may be separated by more amino acids. Alternatively or additionally, the native The amino acid sequence provides one or more amino acid residues of the sulfatase motif sequence. In this case, the selected amino acid residue at the modification site in the Ig heavy chain polypeptide amino acid sequence is Modification to provide a sulfatase motif at the site (e.g., the modification is 1 or The amino acid sequence may be modified (including insertion, deletion, and / or substitution of multiple amino acid residues).

[0399] The antibodies used in the antibody-drug conjugates of the present disclosure may be of various antigen binding specificities. These may include, for example, antigens present on cancer cells, autoimmune diseases, antigens present on cells, antigens present on pathogenic microorganisms, virus-infected cells (e.g., human antigens present on human immunodeficiency virus-infected cells, and antigens present on diseased cells For example, the antibody conjugate may be present on the surface of a cell. The antibody conjugates of the present disclosure have suitable binding affinity. , e.g., 5 x 10 -6 M~10 -7 M, 10 -7 M~5×10 -7 M, 5 x 10 -7 M ~10 -8 M, 10 -8 M~5×10 -8 M, 5 x 10 -8 M~10 -9 M, or 10 -9 It can bind to the antigen with a higher binding affinity than M.

[0400] As a non-limiting example, the subject antibody conjugates may target antigens present on cancer cells (e.g., , tumor-specific antigens, antigens overexpressed on cancer cells, etc.), and The conjugated moiety may be a drug, e.g., a cytotoxic compound (e.g., a cytotoxic small molecule, a cytotoxic agent, a For example, the subject antibody conjugates can be used to treat cancer The conjugated moiety can be specific for an antigen on a cell, and the conjugated moiety can be a drug, e.g., a cytotoxic compound. The compound is a cytotoxic small molecule, a cytotoxic synthetic peptide, or the like.

[0401] As a further non-limiting example, the subject antibody conjugates may be used to express IL-16 on virally infected cells. antigens present in the host (e.g., when the antigen is encoded by a virus; when the antigen is When expressed on a cell type that is more susceptible to infection, the conjugated moiety can bind to the The moiety can be a drug, e.g., a viral fusion inhibitor. The gate can bind to antigens present on virus-infected cells and is conjugated to The moiety can be a drug, for example, a viral fusion inhibitor.

[0402] Drugs for conjugation to polypeptides The present disclosure provides drug-polypeptide conjugates (e.g., antibody-drug conjugates). A number of drugs are suitable for use as reactive partners for conjugation to antibodies. Examples of drugs include: These include small molecule drugs and peptide drugs.

[0403] A "small molecule drug," as used herein, is a compound that exhibits a pharmaceutical activity of interest and and generally a molecular weight of 800 Da or less, or 2000 Da or less refers to compounds, e.g., organic compounds, but can include molecules up to 5 kDa, Small inorganic molecules can be as small as 100 kDa. Small inorganic molecules are molecules that do not contain carbon atoms. A small organic molecule refers to a compound containing at least one carbon atom.

[0404] "Peptide drug," as used herein, refers to an amino acid-containing polymeric compound. refers to naturally occurring and non-naturally occurring peptides, oligopeptides, cyclic peptides, It is meant to encompass peptides, polypeptides, and proteins, as well as peptidomimetics. Peptide drugs may be obtained by chemical synthesis or from genetically encoded sources. Peptide drugs may be produced from various sources (e.g., recombinant sources). The molecular weight can be between 200 Da and 10 kDa or greater. Suitable peptides include cytotoxic peptides, angiogenic peptides, and anti-angiogenic peptides. , B cell activation peptides, T cell activation peptides, antiviral peptides peptides that inhibit viral fusion, peptides that increase the production of one or more lymphocyte populations peptides, antimicrobial peptides, growth factors, growth hormone-releasing factors, vasoactive peptides, Anti-inflammatory peptides, peptides that regulate glucose metabolism, antithrombotic peptides, antinociceptive peptides These include vasodilatory peptides, platelet aggregation inhibitors, and analgesics. Not limited to these.

[0405] Examples of drugs include small molecule drugs, such as cancer chemotherapeutic agents. When the peptide is an antibody (or a fragment thereof) having specificity for tumor cells, the antibody The present invention provides a method for the preparation of a medicament for cancer therapy, comprising the step of conjugating a medicament to a cancer chemotherapeutic agent .... The cancer chemotherapeutic agent can be modified as described in the specification. Non-peptide (i.e., non-proteinaceous) compounds that reduce proliferation of cells Non-limiting examples of chemotherapeutic agents include alkylating agents, cytotoxic agents, and cytostatic agents. Antioxidants, nitrosoureas, antimetabolites, antitumor antibiotics, plant (vinca) alkaloids and steroid hormones. Peptide compounds can also be used. do.

[0406] Suitable cancer chemotherapeutic agents include dolastatins and their active analogs and derivatives; and auristatins and their active analogs and derivatives (e.g., monomethylauristatins, auristatin D (MMAD), monomethyl auristatin E (MMAE), and monomethyl For example, the compounds of the present invention are those disclosed in International Publication No. 96 / 332. 12, WO 96 / 14856, and U.S. Pat. No. 6,323,315 For example, dolastatin 10 or auristatin PE may be used in combination with antibody-drug conjugates of the disclosure. Suitable cancer chemotherapeutic agents also include maytansinolate. ides and their active analogs and derivatives (e.g., EP 1391213, and Li u et al(1996)Proc.Natl.Acad.Sci.USA 93:8 618-8623), duocarmycins and their active analogs and derivatives ( For example, synthetic analogs, including KW-2189 and CB1-TM1, and benzodiazepine and its active analogues and derivatives (e.g., pyrrolobenzodiazepines ( PBD).

[0407] Agents that act to reduce cell proliferation are known in the art and are widely used. Such agents include alkylating agents, such as nitrogen mustard. , nitrosoureas, ethyleneimine derivatives, alkylsulfonates, and triazenes, For example, but not limited to, mechlorethamine, cyclophosphamide (Cytoxan trademark), melphalan (L-sarcolysin), carmustine (BCNU), lomustine (CCNU), semustine (methyl-CCNU), streptozocin, chlorozotocin, Uracil mustard, chlormethine, ifosfamide, chlorambucil, pipobroman, Triethylenemelamine, triethylenethiophosphoramine (triethyleneth iophosphoramine), busulfan, dacarbazine, and temozolomide Examples include:

[0408] Antimetabolites include folic acid analogs, pyrimidine analogs, purine analogs, and adenosine deaminase inhibitors, such as, but not limited to, cytarabine (CYTOSA) RU), cytosine arabinoside, fluorouracil (5-FU), floxuridine ( FudR), 6-thioguanine, 6-mercaptopurine (6-MP), pentostatin, 5-fluorouracil (5-FU), methotrexate, 10-propargyl-5,8- Dideazafolic acid (PDDF, CB3717), 5,8-dideazatetrahydrofolic acid (DDA THF), leucovorin, fludarabine phosphate, pentostatin ine), and gemcitabine.

[0409] Suitable natural products and their derivatives (e.g., vinca alkaloids, antitumor antibiotics, Substances, enzymes, lymphokines, and epipodophyllotoxins include Ara-C, Clitaxel (Taxol®), docetaxel (Taxotere®) )), deoxycoformycin, mitomycin-C, L-asparaginase, azathioprine Phosphorus; Brequinar; Alkaloids, e.g., Vincristine, Vinblastine, Vinorel Vin, vindesine, etc.; podophyllotoxins, e.g., etoposide, teniposide, etc.; anti Biologics, such as anthracyclines, daunorubicin hydrochloride (daunomycin, rubido mycin, cerubicin), idarubicin, doxorubicin, epirubicin and morpholin Derivatives of phenoxizone biscyclopeptides cyclopeptides), such as dactinomycin; basic glycopeptides, such as bleomycin; anthraquinone glycosides, e.g., plicamycin (mithramycin ); anthracenediones, such as mitoxantrone; azirinopyrroloindolediones (azirinopyrrolo indolediones), e.g., mitomycin and macrocyclic immunosuppressants, such as cyclosporine, FK-506 (tacrolimus , Prograf), rapamycin, and the like.

[0410] Other antiproliferative cytotoxic agents include navelbene, CPT-11, and ana Anastrazole, letrozole, capecitabine, reloxafi reloxafine, cyclophosphamide, ifosamide ), and droloxafine.

[0411] Microtubule-interacting agents with antiproliferative activity are also suitable for use, including allocolchicine (N SC 406042), halichondrin B (NSC 609395), colchicine (NS C 757), colchicine derivatives (e.g., NSC 33410), dolstatin 10 ( dolstatin 10) (NSC 376128), maytansine (NSC 153 858), Rhizoxin (NSC 332598), Paclitaxel (Taxol (registered trademark) Taxol® derivatives, docetaxel (Taxotere® ), thiocolchicine (NSC 361792), trityl cysterol (trityl c ysterin), vinblastine sulfate, vincristine sulfate, natural and synthetic epoxy Tirons, including but not limited to eopthilone A; Epothilone B, discodermolide; as well as estramustine, nocodazole, etc. These include, but are not limited to:

[0412] Hormone modulators and steroids (including synthetic analogues) suitable for use Corticosteroids, such as prednisone and dexamethasone; estrogens gens and pregestins, such as hydroxypropyl caproate Gesterone, medroxyprogesterone acetate, megestrol acetate, estradiol, clomiphene, tamoxifen, etc., and adrenal cortical suppressants, e.g., aminoglutethimide 17α-ethinylestradiol; diethylstilbestrol, testosterone , fluoxymesterone, dromostanolone propionate, testolactone, methylpropional Rednisolone, methyl-testosterone, prednisolone, triamcinolone, chlorothion Rianisene, hydroxyprogesterone, aminoglutethimide, estramustine, acetic acid Medroxyprogesterone, leuprolide, flutamide (Drogenil), toremifene ( Fairstone®, and Zoladex®. Estrogen stimulates proliferation and differentiation and therefore binds to estrogen receptors. Compounds that inhibit T cell proliferation are used to block this activity. It can be inhibited.

[0413] Other suitable chemotherapeutic agents include metal complexes, e.g., cisplatin (cis-DDP), carboplatin, etc.; ureas, e.g., hydroxyurea, and hydrazines, e.g., N- Methylhydrazine; epidophyllotoxin; topo Isomerase inhibitors; procarbazine; mitoxantrone; leucovorin; tegafur, etc. Other antiproliferative agents of interest include immunosuppressants, e.g., mycophenone. thalidomide, desoxyspergualin , azasporine, leflunomide, mizoribine, azaspirane ( SKF 105685; Iressa® (ZD 1839, 4-(3-chloro) 3-(4-morpholinyl)-7-methoxy-6-(3-(4-fluorophenylamino)propanol) hydroxyquinazoline).

[0414] Taxanes are suitable for use. "Taxane" refers to any of the following in addition to paclitaxel: "Paclitaxel" (as used herein) includes active taxane derivatives or prodrugs. and analogs, formulations, and derivatives, such as docetaxel, TAXOL™, TA XOTERE™ (a formulation of docetaxel), a 10-desacetylated form of paclitaxel 3'N-desbenzoyl-3'Nt-butoxycarbonyl esters of chloroquine and paclitaxel It should be understood that the present invention includes the use of techniques known to those skilled in the art. It can be easily prepared (WO 94 / 07882, WO 94 / 0788 1, WO 94 / 07880, WO 94 / 07876, WO 93 / 23555, WO 93 / 10076; U.S. Pat. No. 5,294,637 Specification No. 5,283,253, Specification No. 5,279,949, No. 5,27 Specification No. 4,137, Specification No. 5,202,448, Specification No. 5,200,534 EP 590,267; see also EP 590,267; or Various commercial sources, e.g., Sigma Chemical Co., St. Louis, MO. s, Mo. (T7402 from Taxus brevifolia; or Taxus yannanensis (T-1912).

[0415] Paclitaxel is not only the common chemically available form of paclitaxel; Analogs and derivatives (e.g., Taxotere™ docetaxel, as described above) ) and paclitaxel conjugates (e.g., paclitaxel-PEG, paclitaxel It should be understood that the term also refers to paclitaxel-dextran, paclitaxel-xylose, It is.

[0416] A variety of known derivatives, including both hydrophilic and hydrophobic derivatives, are also known as "taxa." Taxane derivatives are included in the term "taxanes." galactose and mannose derivatives as described in WO 99 / 14209 piperazino and other derivatives as described in WO 99 / 0 9021, WO 98 / 22451, and U.S. Pat. No. 5,869,680. taxane derivatives as described in the specification, 6-thio derivatives as described in WO 98 / 28288; Sulfenamide derivatives described in U.S. Pat. No. 5,821,263, and U.S. Taxol derivatives described in Japanese Patent No. 5,415,869 are included, but are not limited to these. It is not limited to prodrugs of paclitaxel, for example, but not limited to, International Publication No. 98 / 58927, International Publication No. 98 / 13059, and U.S. Pat. Further, the present invention includes those described in US Pat. No. 2,470,701.

[0417] Suitable biological response modifiers for use include: (1) tyrosine kinases (RTKs) (2) inhibitors of serine / threonine kinase activity; (3) inhibitors of tumor-associated antigen activity. (4) apoptosis receptor antagonists, e.g., antibodies that specifically bind to tumor antigens; agonist, (5) interleukin-2, (6) IFN-α, (7) IFN-γ, (8) (9) inhibitors of angiogenesis, including, but not limited to, colony-stimulating factors .

[0418] Examples of drugs include small molecule drugs, such as cancer chemotherapeutic agents. When the peptide is an antibody (or a fragment thereof) having specificity for tumor cells, the antibody , modified chemotherapeutic agents, such as microtubule-interacting agents, which can then be conjugated to cancer chemotherapeutic agents. It can be modified as described herein to include certain amino acids. In embodiments, the drug is a microtubule-interacting agent with antiproliferative activity, e.g., a maytansinoid. In certain embodiments, the drug is a maytansinoid, and the maytansinoid is The following structure: [ka]

[0419] and [ka]

[0420] is the moiety of the maytansinoid and second maytansinoid in the conjugates and compounds described herein. Linker, L 2 The "connection point" indicates the connection point between the [ka]

[0421] The symbols represent the N and N of the maytansinoid in the conjugates and compounds described herein. and a second linker, L 2 For example, in formula (I), W 1 may be a maytansinoid, for example, a maytansinoid of the structure shown above. Ku, [ka]

[0422] is a linker comprising a maytansinoid and a second linker, L 2 In some cases, In some cases, maytansinoids of the above structure are sometimes referred to as deacylmaytansines.

[0423] In certain embodiments, the drug is an antimitotic agent, e.g., an auristatin or an active ingredient thereof. auristatin analogs or derivatives (e.g., monomethylauristatin D (MM AD), monomethylauristatin E (MMAE), and monomethylauristatin F (MMAF), etc. In certain embodiments, the drug has the following structure: [ka]

[0424] It is an MMAE with the following properties.

[0425] For example, MMAE activators include: [ka]

[0426] and the antibody-drug conjugate may be included in the antibody-drug conjugate as follows: [ka]

[0427] The auristatin and second ligand in the conjugates and compounds described herein L. 2 For example, [ka]

[0428] The symbol may refer to, for example, the N of an auristatin and the second linker, as shown in formula (I). -L 2 For example, in formula (I), W 1 are auristatins, e.g. For example, it can be MMAE, and in the above structure [ka]

[0429] is a linker consisting of MMAE and a second linker, L 2 It points to the connection point between

[0430] In certain embodiments, the drug is a DNA alkylating agent, e.g., a duocarmycin. Examples of ducarmycins include duocarmycin A , Duocarmycin B1, Duocarmycin B2, Duocarmycin C1, Duocarmycin Calmycin C2, Duocarmycin D, Duocarmycin SA, and CC-10 65. In some embodiments, the duocarmycins Duocarmycin analogues, including but not limited to, adozelesin, bizelesin , or carzelesin.

[0431] In some instances, the duocarmycin has the following structure: [ka]

[0432] It is a compound having the formula:

[0433] For example, the duocarmycin activator may be: [ka]

[0434] or [ka]

[0435] and the antibody-drug conjugate may be included in the antibody-drug conjugate as follows: [ka]

[0436] is the moiety of ducarmycin and the second moiety in the conjugates and compounds described herein. Linker, L 2 For example, [ka]

[0437] The symbol may represent, for example, a duocarmycin and a second linker, as shown in formula (I). -L 2 For example, in formula (I), W 1 is duocarmycin , for example, ducarmycin as shown above, [ka]

[0438] is a duocarmycin and a second linker, L 2 It points to the connection point between

[0439] As noted above, in certain embodiments, L 2 is the formula -(L 21 ) e -(L 22 ) f - (L 23 ) g -(L 24 ) h- a second linker described by L 21 , L 22 , L 23 and L 24 are each independently a second linker subunit. In the embodiment, L 21 is attached to the first severable portion. In terms of form, L 21 If there is a W 1 Attached to (drugs). In certain embodiments, L 22 If there is a W 1 Attached to (drugs) In certain embodiments, L 23 If there is a W 1 (drug) In certain embodiments, L 24 If there is a W 1 (drug) It is being used.

[0440] As noted above, in certain embodiments, the second linker -(L 21 ) e -(L 22 ) f -(L 23 ) g -(L 24 ) h - is the formula -(T 5 -V 5 ) e -(T 6 -V 6 ) f -(T 7 -V 7 ) g -(T 8 -V 8 ) h - wherein e, f, g and h are each independently and the sum of e, f, g and h is 0 to 4. As mentioned above, L 21 is attached to the first detachable portion. In certain embodiments, T 5 is attached to the first severable portion. In an embodiment, V 5 is W 1 In certain embodiments, the above As mentioned above, L 22 If there is a W 1 (drugs) attached. Therefore, in certain embodiments, T 6 If there is a W 1 (drug) or V 6 If there is a W 1 (drug) attached to a particular In certain embodiments, as described above, L 23 If there is a W 1 (drug) Therefore, in certain embodiments, T 7 If there is a W 1 ( Drug) or V 7 If there is a W 1 (drug) In certain embodiments, as described above, L 24 If there is a W 1 (drug). Therefore, in certain embodiments, T 8 There exists If so, it is W 1 (drug) or V 8 If there is a W 1 Attached to (drugs).

[0441] Embodiments of the disclosure include those in which the antibody is coupled to one or more drug moieties, e.g., two drug moieties, three drug moieties, or 1 drug portion, 4 drug portions, 5 drug portions, 6 drug portions, 7 drug portions, 8 drug portions conjugated to 1 drug moiety, 9 drug moieties, or 10 or more drug moieties The drug moiety may be one or more conjugates in the antibody, as described herein. In certain embodiments, the conjugate may be conjugated to the antibody at the site , 0.1 to 10, or 0.5 to 10, or 1 to 10, e.g., 1 to 9, or 1 to 8 , or 1-7, or 1-6, or 1-5, or 1-4, or 1-3, or 1 In certain embodiments, the antibody has an average drug-to-antibody ratio (DAR) (molar ratio) in the range of 0.1 to 2. The conjugate is 1 to 2, e.g., 1, 1.1, 1.2, 1.3, 1.4, 1.5 , 1.6, 1.7, 1.8, 1.9 or 2. In certain embodiments, the conjugate has an average DAR of 1 to 1.5. The conjugates have an average DAR of 1.5 to 2. By average is meant the arithmetic mean.

[0442] The drug conjugated to the polypeptide must have a reactive partner for reaction with the polypeptide. If the drug is a peptide drug, it may be modified to incorporate a reactive moiety (e.g. , aminooxy or hydrazide can be used in the N-terminal region, N-terminus, C-terminal region, C-terminus of a peptide. The aminooxy group can be located at a terminal or internal position. In this example, the peptide is Boc-protected. The amino group of the peptide is converted to a carboxylic acid group and an oxy-N-Boc For example, the amino group of a peptide can be reacted with a compound containing a 3-(2, 5-dioxopyrrolidin-1-yloxy)propanoic acid. Other variations of the compounds containing the oxy-N protecting group include different numbers of alkylene linkers. The amino acid sequence of the peptide may include substituents on the carbon atoms in the amino group and on the alkylene linker. Reactions between the carboxylate group and compounds containing carboxylic acid and oxy-N protecting groups are carried out using standard phenolics. This occurs through peptide coupling chemistry. Examples of drugs include DCC (dicyclohexylcarbodiimide), DIC (diisopropyl Carbodiimide), di-p-toluoylcarbodiimide rbodiimide), BDP (1-benzotriazole diethyl phosphate-1- Cyclohexyl-3-(2-morpholinylethyl)carbodiimide), EDC (1-(3 -dimethylaminopropyl-3-ethyl-carbodiimide hydrochloride), cyanofluoride Nur, cyanuric chloride, TFFH (tetramethylfluoroformamidinium hexafluoro) Tetramethyl fluoroformamide m hexafluorophosphosphate), DPPA (diphenylphosphosphate) diphenylphosphorazidate), BOP (Ben 1-isotriazolyloxytris(dimethylamino)phosphonium hexafluoro phosphate), HBTU (O-benzotriazol-1-yl-N,N,N',N'- Tetramethyluronium hexafluorophosphate), TBTU (O-benzotriazo 1-yl-N,N,N',N'-tetramethyluronium tetrafluoroborate ), TSTU (O-(N-succinimidyl)-N,N,N',N'-tetramethyluron ammonium tetrafluoroborate), HATU(N-[(dimethylamino)-1-H-1, 2,3-Triazolo[4,5,6]-pyridin-1-ylmethylene]-N-methylmeth ammonium hexafluorophosphate N-oxide), BOP-Cl (bis(2-hydroxybenzoate) PyBOP ((1-H-1,2-dioxo-3-oxazolidinyl)phosphinic acid chloride), ,3-benzotriazol-1-yloxy)-tris(pyrrolidino)phosphonium tetraoxide trifluorophosphate), BrOP (bromotris(dimethylamino)phosphonium Diethoxyfluorophosphate), DEPBT (3-(diethoxyphosphoryloxy)-1, 2,3-Benzotriazin-4(3H)-one)PyBrOP (Bromotris(pyrrolidine) Examples of suitable phosphonium hexafluorophosphates include, but are not limited to, phosphonium hexafluorophosphate. As a non-limiting example, HOBt and DIC are used as peptide coupling reagents. It is possible.

[0443] Deprotection to expose the amino-oxy functionality is performed on peptides containing N-protecting groups. For example, deprotection of N-oxysuccinimide groups is performed using standard methods for cyclic amide groups. Deprotection conditions are described in Greene and Wuts, Protective Groups in Organic Chemistry,3 rd Ed., 1999, John Wiley & Sons, NY and Harrison Certain deprotection conditions can be found in [Dr.]n et al. The deprotection of the Boc protecting group is performed using TFA. Other reagents for deprotection include hydrazine, methylhydrazine, Examples include phenylhydrazine, sodium borohydride, and methylamine. Products and intermediates may be purified by conventional means, for example, HPLC purification. It can be further refined.

[0444] pH and steric hindrance (i.e., amino acid resistance to reaction with the reactive partner of interest) Factors such as the accessibility of the acid residues are important and the reaction conditions must be carefully considered to provide optimal conjugation conditions. Modification of reaction conditions is well within the skill of one in the art and is routine in the art. Those skilled in the art will recognize that conjugation can be performed with polypeptides present in or on living cells. When carried out, the conditions are selected to be physiologically compatible. for a period of time sufficient for incubation but within a period that is tolerated by the cells (e.g., approximately 30 minutes to 1 hour). The pH can be temporarily lowered over a period of time. The physiological conditions for this are the ketone-azide modification of cells with cell surface azides. The reaction may be similar to that used in the amide reaction (see, for example, U.S. Pat. No. 6,577,627). (See specification No. 0,040).

[0445] Serve as reactive partners with the compounds or conjugates disclosed herein. Small molecule compounds containing, or modified to contain, an alpha nucleophilic group are also disclosed herein. Contemplated for use as a drug in the polypeptide-drug conjugates shown. General methods for chemical synthesis schemes and conditions useful for synthesizing compounds of interest Methods are known in the art (e.g., Smith and March, rch's Advanced Organic Chemistry:Reactio ns,Mechanisms,and Structure,Fifth Editio n, Wiley-Interscience, 2001; or Vogel, A Tex tbook of Practical Organic Chemistry, Inc. luding Qualitative Organic Analysis,Four th Edition, New York: Longman, 1978).

[0446] formulation The conjugates of the present disclosure can be formulated in a variety of different ways. When the conjugate is an antibody-drug conjugate, the conjugate comprises a drug, an antibody and formulated in a manner compatible with the condition to be treated and the route of administration to be used. do.

[0447] In some embodiments, any conjugate of the present disclosure and a pharmaceutically acceptable excipient A pharmaceutical composition comprising:

[0448] The conjugate (e.g., antibody-drug conjugate) may be in any suitable form, e.g., , can be provided in the form of a pharmaceutically acceptable salt and can be administered by any suitable route of administration, e.g. The conjugate can be formulated for oral, topical or parenteral administration. Injectables (such as those in embodiments where they are administered intravenously or directly into tissue) ), the conjugate may be provided as a ready-to-use dosage form or A reconfigurable, shelf-stable powder or formulation consisting of pharmaceutically acceptable carriers and excipients. It can be provided as a liquid.

[0449] Methods for formulating conjugates can be adapted from readily available For example, the conjugate may be prepared by combining a therapeutically effective amount of the conjugate and a pharmaceutically acceptable carrier. It may be provided in a pharmaceutical composition comprising a carrier (e.g., saline). may contain other additives such as buffers, stabilizers, and preservatives. In this embodiment, the formulation is suitable for administration to a mammal, e.g., for administration to a human. It is suitable for

[0450] Treatment method The antibody-drug conjugates of the present disclosure are prepared by subjecting the parent drug (i.e., the drug prior to conjugation to the antibody) to a It has use in treating a condition or disease in a subject suitable for treatment by administration.

[0451] In some embodiments, a subject is administered an effective amount (e.g., a therapeutically effective amount) of any of the conjugates of the present disclosure. A method is provided that includes administering a gate.

[0452] In certain aspects, a method of delivering a drug to a target site in a subject comprises administering to the subject administering a pharmaceutical composition comprising any of the disclosed conjugates, wherein administering to release a therapeutically effective amount of the drug from the conjugate at a target site in a subject. For example, methods are provided for using antibodies of the present disclosure, as described herein, that are effective in treating a patient with an antibody- The drug conjugate may comprise a cleavable linker, e.g., a first cleavable moiety and a second cleavable moiety. The linker may be an enzymatically cleavable linker or a chemically cleavable linker. The second cleavable moiety can be cleaved to prevent cleavage of the first cleavable moiety. In some instances, the cleavable linker is cleaved under appropriate conditions to release the desired target moiety of the drug. The drug can be separated or released from the antibody at the first cleavable site. The second cleavable linker protects the first cleavable linker from cleavage. It may be cut to allow for cutting into two or more parts. resulting in cleavage of the cleavable linker to the antibody- The drug is released from the drug conjugate.

[0453] In certain embodiments, the first cleavable moiety is an enzymatically cleavable moiety. In some cases, the enzyme that promotes cleavage of the first cleavable moiety is An enzyme that is administered to a subject (i.e., exogenous to the subject being treated). For example, the first enzyme may be administered prior to, concurrently with, or after administration of the antibody-drug conjugate described herein. can be administered later.

[0454] In certain embodiments, the second cleavable moiety is an enzymatically cleavable moiety. In some cases, the enzyme that promotes cleavage of the second cleavable moiety is the target of treatment. An enzyme that is administered to a subject (i.e., exogenous to the subject being treated). For example, the second enzyme may be administered prior to, concurrently with, or after the administration of the antibody-drug conjugate described herein. can be administered later. In certain embodiments, the first enzyme and the second enzyme are different enzymes.

[0455] In other cases, the first enzyme that promotes cleavage of the first cleavable moiety is a target of the subject to be treated. (i.e., endogenous to the subject to be treated). For example, the first enzyme is present at the desired site of action of the drug of the antibody-drug conjugate. The antibody of the antibody-drug conjugate may be specifically targeted to a desired site of action. It may be (e.g., it may specifically bind to an antigen present at the desired site of action) The site of action of also includes the presence of a first enzyme. In some cases, the first enzyme is present in the target cell. It is present in excess at the desired site of action compared to other compartments in the body of the subject. For example, the first enzyme may be located at a desired site of action relative to other compartments in the body of the subject to be treated. In some cases, the first enzyme may be overexpressed in a particular compartment or location. Due to the localization of the first enzyme in the target site, it is present in excess at the desired site of action. For example, the first enzyme may be associated with a particular structure within the desired site of action, e.g., a lysosome. In some cases, the first enzyme may be present in a lysosomal compartment relative to other compartments in the subject's body. In some embodiments, the lysosome containing the first enzyme is present in excess in the lysosome. - the desired site of action of the drug of the drug conjugate, e.g., the site to be treated with the drug In certain embodiments, the first enzyme is a protein. ases, for example, human protease enzymes (e.g., cathepsin B).

[0456] In certain embodiments, the second enzyme that facilitates cleavage of the second cleavable moiety is an enzyme that is present in the subject to be treated (i.e., endogenous to the subject to be treated) For example, the second enzyme may act at the desired site of action of the drug in the antibody-drug conjugate. The antibody of the antibody-drug conjugate may be specifically targeted to the desired site of action. It may be modified (e.g., it may specifically bind to an antigen present at the desired site of action). In some cases, the desired site of action also includes the presence of a second enzyme. In some cases, the second enzyme is present in excess at the desired site of action compared to other compartments in the body of the subject to be treated For example, the second enzyme may be present in a desired compartment relative to other compartments in the body of the subject being treated. In some cases, the second enzyme may be overexpressed at the site of action. Due to the localization of the second enzyme in a region or location, it may be present in excess at the desired site of action. For example, the second enzyme may be located in a specific structure within the desired site of action, e.g., a lysosome. In some cases, the second enzyme may be associated with a different enzyme than another compartment in the subject's body. In some embodiments, the lysosomes containing the second enzyme are present in excess. The antibody-drug conjugate is used to treat a subject with a desired site of action of the drug, e.g. In certain embodiments, the second enzyme is found at the site of the cancer or tumor to be treated. The enzyme is a glucuronidase, e.g., a human glucuronidase enzyme (e.g., lysosomal β-glucuronidase). glucuronidase).

[0457] First cleavable moiety and second cleavage of the antibody-drug conjugates described herein Any suitable enzyme can be used for cleaving the possible portions. Upon cleavage of the first cleavable moiety and the second cleavable moiety of the antibody-drug conjugate, Other enzymes for use in other vertebrates (e.g., spirits), including, but not limited to, Enzymes from long animals (mouse, rat, cat, pig, quail, goat, dog, etc.) are also suitable. This happens sometimes.

[0458] In certain embodiments, the antibody-drug conjugate is substantially stable under standard conditions. By substantially stable, the cleavable linker of the antibody-drug conjugate is It is known that no significant amount of cleavage occurs in the absence of the first enzyme and the second enzyme. For example, as described above, the second cleavable moiety can be used to cleave the first cleavable moiety. The cleavable linker in the antibody-drug conjugate can be protected from cleavage. , which does not undergo significant amounts of cleavage in the absence of a second enzyme, as described above. The cleavable linker of the antibody-drug conjugate may be 25% or less of the antibody-drug conjugate. less than, for example, 20% or less, or 15% or less, or 10% or less % or less, or 5% or less, or 4% or less, and is 3% or less, or 2% or less, or 1% or less Substantially stable to cleavage in the absence of the first enzyme and / or the second enzyme. In some cases, the antibody-drug conjugate may be The cleavable linker undergoes significant cleavage in the absence of the first enzyme and / or the second enzyme. but can be cleaved in the presence of the first enzyme and the second enzyme For example, the antibody-drug conjugate is substantially stable after administration to a subject. In some cases, the antibody-drug conjugate can be administered to a subject. and subsequently the antibody-drug conjugate is substantially stable after administration at the desired site of action. and in the presence of a second enzyme, the second cleavable moiety is released from the cleavable linker. cleaved to expose a first cleavable moiety for subsequent cleavage by a first enzyme This then releases the drug at the desired site of action. In this embodiment, after administration to a subject, the antibody-drug conjugate binds to the first enzyme and / or the second enzyme. In the absence of enzyme for an extended period of time, e.g., 1 hour or more, or 2 hours or longer, or 3 hours or longer, or 4 hours or longer or longer, or 5 hours or longer, or 6 hours or longer, or or 7 hours or more, or 8 hours or more, or 9 hours or longer, or 10 hours or longer, or 15 hours or longer or longer, or 20 hours or longer, or 24 hours (1 day) or longer or longer, or 2 days or longer, or 3 days or longer, or 4 days or more, or 5 days or more, or 6 days or more Stable for 7 days (1 week) or longer. In embodiments, the antibody-drug conjugate is a non- In the presence of a range of pH values, e.g., 2 to 10, or 3, for an extended period of time. It is stable at a pH range of 0.01 to 9, or 4 to 8, or 5 to 7, or 6 to 7.

[0459] As noted above, the antibody-drug conjugates of the present disclosure are suitable for treatment by administration of the parent drug. "Treatment" has utility in treating a condition or disease in a subject. It is meant that at least remission of symptoms associated with the affected condition is achieved, and remission is , a small change in a parameter, e.g., a symptom scale associated with the condition being treated. Treatment is therefore also used in the broad sense to refer to at least the reduction of pathological The condition, or at least the symptoms associated with it, are completely inhibited, e.g., prevented from occurring. The host is prevented or stopped, e.g., terminated, thereby preventing the host from the condition or the Therefore, treatment includes the situation where the patient no longer suffers from at least the symptoms that characterize the condition. ) prevention, i.e., reducing the risk of clinical symptoms developing, e.g., preventing clinical symptoms from developing (ii) inhibiting, i.e., preventing, the progression of a disease to a deleterious state; arrest of the onset or further development of symptoms, e.g., reduction or complete inhibition of active disease; and / or (iii) palliation, i.e., causing regression of clinical symptoms.

[0460] The subject to be treated can be a subject in need of therapy, are suitable subjects for treatment with the parent drug. Thus, various subjects are Generally, such subjects may be suitable for treatment using the antibody-drug conjugates described above. "Mammals" and humans are the subjects of interest. Other subjects include domestic pets (e.g. animals (e.g., dogs and cats), livestock animals (e.g., cows, pigs, goats, and horses), rodents Animals (e.g., mice, guinea pigs, and rats, e.g., in animal models of disease) In addition to mammals (e.g., chimpanzees and monkeys), non-human primates can also be included. do.

[0461] The amount of antibody-drug conjugate administered may be adjusted based on the dose and / or dosing regimen of the parent drug. The initial decision can be made based on the guidance of the antibody-drug conjugate. The carboxylate provides targeted delivery and / or enhanced serum half-life of the conjugated drug. Therefore, at least one of a dose reduction or a reduction in administration in the dosing regimen may be performed. Thus, the antibody-drug conjugate can provide the antibody-drug of the present disclosure. Dose reduction and / or or a reduced dose may be provided.

[0462] Furthermore, as mentioned above, antibody-drug conjugates allow for control of the stoichiometry of drug delivery. Therefore, the dosage of the antibody-drug conjugate can be The dose can be calculated based on the number of drug molecules provided per dose.

[0463] In some embodiments, multiple doses of the antibody-drug conjugate are administered. The frequency of administration of the monoclonal conjugate can depend on any of a variety of factors, e.g., the severity of the symptoms, the patient's For example, in some embodiments, the antibody-drug ratio may vary depending on the condition of the antibody. Conjugates are monthly, twice monthly, three times monthly, every other week, once a week (qwk), twice a week, and three times a week. , 4 times a week, 5 times a week, 6 times a week, every other day, every other day (qd / od), twice a day (bds / bi d), or three times a day (tds / tid).

[0464] Cancer treatment methods The present disclosure provides a method comprising delivering a conjugate of the present disclosure to an individual with cancer. The method provides a method for treating a variety of cancers, including carcinomas, sarcomas, leukemias, and lymphomas. In the context of cancer, the term "treatment" includes reducing the growth of solid tumors, Inhibition of cancer cell replication, reduction of overall tumor burden, and the development of cancer-associated 1 or Including one or more (e.g., each) of the multiple symptom amelioration.

[0465] Carcinomas that can be treated using the subject method include esophageal carcinoma, hepatocellular carcinoma, basal cell carcinoma, and esophageal carcinoma. Cancer (form of skin cancer), squamous cell carcinoma (various tissues), bladder cancer, e.g., transitional cell carcinoma (bladder malignant neoplasms), bronchogenic carcinoma, colon cancer, colorectal cancer, gastric cancer, lung cancer, e.g., small cell lung cancer Cancer and non-small cell cancer, adrenocortical cancer, thyroid cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, adenocarcinoma, Sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, renal cell carcinoma, intraductal carcinoma or biliary Ductal carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, uterine cancer, testicular cancer, osteogenic These include, but are not limited to, uterine carcinoma, epithelial carcinoma, and nasopharyngeal carcinoma.

[0466] Sarcomas that can be treated using the subject method include fibrosarcoma, myxosarcoma, and liposarcoma. sarcoma, chondrosarcoma, chordoma, osteogenic sarcoma, osteosarcoma, angiosarcoma, endothelial sarcoma, lymphangiosarcoma, lymphoma Endothelioma, synovium, mesothelioma, Ewing's sarcoma, leiomyosarcoma, rhabdomyosarcoma, and other soft tissue These include, but are not limited to, sarcoma of the tissue.

[0467] Other solid tumors that can be treated using the subject method include gliomas, astrocytic tumors, and gliomas. tumor, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma These include, but are not limited to, melanoma, neuroblastoma, and retinoblastoma.

[0468] Leukemias that can be treated using the subject method include: a) chronic myeloproliferative syndromes (a neoplastic disorder of multipotential hematopoietic stem cells), b) acute myeloid Myeloid leukemia (neoplastic transformation of multipotent hematopoietic stem cells or hematopoietic cells with restricted lineage potential, c) Chronic lymphocytic leukemia (CLL; a cluster of immunologically immature and functionally incompetent small lymphocytes) Lone proliferation), e.g., B-cell CLL, T-cell CLL, prolymphocytic leukemia, and hairy leukemia, and d) acute lymphoblastic leukemia (characterized by the accumulation of lymphoblasts) These include, but are not limited to, conditions that can be treated using the subject methods. Lymphomas include B-cell lymphomas (e.g., Burkitt lymphoma), Hodgkin lymphoma, and non-Hodgkin's B-cell lymphoma.

[0469] In certain aspects, a method of treating cancer in a subject comprises administering to the subject a therapeutically effective amount of administering a pharmaceutical composition comprising any of the conjugates of the present disclosure, Methods are provided that are effective for treating cancer in a subject.

[0470] [Example] The following examples provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention. It is described for the purpose of illustration and is not intended to limit the scope of what the inventors regard as their invention. The experiments below are not intended to be representative of all or the only experiments performed. Every effort has been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.). Experimental results were obtained, but some experimental error and deviations should be considered. If not, parts are parts by weight, molecular weight is weight average molecular weight, and temperature is in degrees Celsius. , pressure is at or near atmospheric pressure. By "average" is meant the arithmetic mean. Abbreviations, e.g., bp, base pair(s); kb, kilobase(s); pl, picoliters; s or sec, seconds; mi n, minute(s); h or hr, hour(s); aa, (one or more) (multiple) amino acids; kb, (one or more) kilobases; bp, (one or more) bases pair; nt, (one or more) nucleotides; im, intramuscular (intramuscularly); ip ., intraperitoneal (into the abdominal cavity); and sc, subcutaneous (under the skin), etc. may be used. be.

[0471] General synthetic procedure Generally known chemical synthetic schemes and conditions useful for synthesizing the disclosed compounds Many general references are available (e.g., Smith and Ma rch,March's Advanced Organic Chemistry:R eactions,Mechanisms,and Structure,Fifth Edition, Wiley-Interscience, 2001; or Vogel ,A Textbook of Practical Organic Chemist ry,Including Qualitative Organic Analysis s, Fourth Edition, New York: Longman, 1978. see).

[0472] Compounds as described herein can be purified using any purification protocol known in the art. Call, e.g., chromatography, e.g., HPLC, preparative thin layer chromatography , purified by flash column chromatography and ion exchange chromatography Any suitable stationary phase can be used, including normal and reverse phase as well as ionic resins. In certain embodiments, the disclosed compounds can be used on silica gel and and / or alumina chromatography. tion to Modern Liquid Chromatography,2nd Edition, ed.LRSnyder and JJKirkland, John Wiley and Sons, 1979; and Thin Layer C chromatography, ed E. Stahl, Springer-Verlag ,New York,1969.

[0473] During any process for the preparation of the subject compounds, sensitive or reactive groups on any molecule involved may be removed. It may be necessary and / or desirable to protect the JFWMcOmie,''Protective Groups in Orga nic Chemistry'', Plenum Press, London and New York 1973, TWGreene and PGMWuts, ''Protective Groups in Organic Synthesis '',Third edition,Wiley,New York 1999,''T he Peptides'';Volume 3(editors:E.Gross a nd J. Meienhofer), Academic Press, London a nd New York 1981, ''Methoden der organisc hen Chemie'', Houben-Weyl,4 th edition,Vol .15 / l,Georg Thieme Verlag,Stuttgart 1974 , H.-D. Jakubke and H. Jescheit,''Aminosaur en,Peptide,Proteine'',Verlag Chemie,Wein heim, Deerfield Beach, and Basel 1982, and / or Jochen Lehmann,''Chemie der Kohlenhyd rate:Monosaccharide and Derivate'',Georg Conventional methods such as those described in Thieme Verlag, Stuttgart 1974 The protecting groups can be added using methods known in the art. The oxidized material may be removed at a later stage, where convenient.

[0474] The subject compounds are prepared from commercially available starting materials and / or by conventional synthetic methods. The compounds can be synthesized via a variety of different synthetic routes using starting materials as described herein. Various examples of synthetic routes that can be used to synthesize the compounds disclosed in the document are provided below. This is depicted in the scheme:

[0475] Synthesis reagents were obtained from Sigma-Aldrich, Acros, or other commercial sources. Anhydrous solvents were obtained in sealed bottles from commercial sources. Dibromo-α-D-glucuronic acid methyl ester 1, MMAE 8, 17, 46, 57, and 62 were purchased from commercial sources. All reactions were carried out under N2 unless otherwise noted. All experiments were carried out in flame-dried glassware. The solvent was Buchi V-70 0 by vacuum using a Buchi Rotovapor R-114 equipped with a vacuum pump Column chromatography was performed using a Biotage Isolera Prime Preparative HPLC purification was performed using Phenomenex Kine Water with a tex 5μm EVO C18 150 x 21.2mm column s Preparative HPLC unit was used. HPLC analysis was performed using water and 0.1% formic acid. Model G13 at room temperature using a 10-100% gradient of acetonitrile containing 22A Degasser, Model G1311A Quarternary Pu mp, Model G1329A Autosampler, Model G1314 Variable Wavelength Detector, Agilent Por Agile with oshell 120 SB C18, 4.6mm x 50mm column The analysis was performed on an 1100 Series Analytical HPLC. C was monitored at 254 nm.

[0476] [Example 1] Synthesis of MMAE construct 13 The monomethyl auristatin E (MMAE) construct was prepared according to Scheme 1 shown below. I synthesized it like this.

[0477] Scheme 1 [ka]

[0478] TIFF2024170512000084.tif197166

[0479] TIFF2024170512000085.tif40168

[0480] (2S,3R,4S,5S,6S)-2-(5-formyl-2-nitrophenoxy)- 6-(Methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyltriazol- Preparation of acetate (2) Methyl acetobromo-α-D-glucopyranuronate in acetonitrile (100 mL) Ester 1 (6.3 g, 15.9 mmol) and 3-hydroxy-4-nitrobenzaldehyde A solution of aldehyde (0.9 g, 5.4 mmol) in AgO (10.0 g, 43.1 mmol) l) was added. The reaction mixture was stirred in the dark at room temperature for 60 h. The reaction mixture was then concentrated under vacuum. The mixture was condensed and purified using column chromatography (hexane / EtOAc, 9:1 to 1:9 v / v). The residue was purified using HCl to give compound 2 (2.1 g, 81%) as an off-white solid.

[0481] Calculated value: [M+Na] + (C 20 H 21 NNaO 13 ) m / z = 506.1; ESI Actual value: [M+Na] + (C 20 H 21 NNaO 13 ) m / z = 505.7.

[0482] (2S,3R,4S,5S,6S)-2-(2-amino-5-(hydroxymethyl)furan) (phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-trimethylsilyl Preparation of methyltriacetate (3) A solution of compound 2 (460 mg, 0.95 mmol) in EtOAc (15 mL) was treated with Pd / C (10 wt%, 20 mg) and triethylamine (20 μL, 0.14 mmol) The flask was then evacuated and hydrogen from the balloon was added in three repeated cycles. The reaction was stirred vigorously at room temperature for 24 h with a H balloon attached. After removing the catalyst by filtration through a pad of Celite, the filtrate was concentrated in vacuo. The material was dried under high vacuum for 1 h to give a white solid (425 mg, 99%), which was further purified. Used directly for the next step without further purification.

[0483] Calculated value: [M+H] + (C 20 H 26 NO 11 ) m / z = 456.2; ESI measured value :[M+H] + (C 20 H 26 NO 11 ) m / z=455.6 (2S,3R,4S,5S,6S)-2-(2-(((S)-2-((((9H-fluo 9-phenyl)methoxy)carbonyl)amino)propanamido)-5-(hydroxy Methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3, Preparation of 4,5-triyltriacetate (4) Fmoc-Ala-OH (4) in DCM (0.9 mL) and MeOH (0.1 mL) A mixture of crude intermediate 3 (425 mg, 0.93 mmol) and crude intermediate 4 (425 mg, 0.93 mmol) The compound was N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ, 390 mg, 1.58 mmol) was added. The reaction was stirred at room temperature for 1 h. The solvent was removed under vacuum. After removing the HCl, column chromatography (hexane / EtOAc, 9:1 to 1:9 v / The residue was purified using HCl (v) to give compound 4 (0.64 g, 90% in two steps) as a white solid. Got it as a body.

[0484] Calculated value: [M+H] + (C 38 H41 N2O 14 ) m / z = 749.3; ESI measurement Value: [M+H] + (C 38 H 41 N2O 14 ) m / z = 748.7.

[0485] (2S,3R,4S,5S,6S)-2-(2-((S)-2-aminopropanamide )-5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro Preparation of 2H-pyran-3,4,5-triyl triacetate (5) A mixture of compound 4 (640 mg, 0.86 mmol) in MeOH (15 mL) was treated with Pd In three repeated cycles, the flask was then The reaction mixture was evacuated to a temperature of 100°C and filled with H2 gas from a balloon. The mixture was vigorously stirred at room temperature for 3 days. The catalyst was removed by filtration through a pad of Celite. The solution was concentrated under vacuum. The eluent was acetonitrile-water (0.05% TFA, 5-7 The residue was purified by reverse phase chromatography using 0% ethanol. The fractions were pooled and concentrated in vacuo to give compound 5 (341 mg, 76%) as a white solid. Got it.

[0486] Calculated value: [M+H] + (C 23 H 31 N2O 12 ) m / z = 527.2; ESI measurement Value: [M+H] + (C 23 H 31 N2O 12 ) m / z = 526.8.

[0487] (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-((( (9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanal Amido)propanamido)-5-(hydroxymethyl)phenoxy)-6-(methoxycarboxamide Preparation of (2H-pyran-3,4,5-triyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (6) Made Amine 5 (255 mg, 0.49 mmol) and Fmoc- A solution of phosphorus-OH (170 mg, 0.50 mmol) was added to DIPEA (0.15 mL, 0. 86 mmol) and PyAOP (260 mg, 0.50 mmol) were added to the reaction mixture. The mixture was stirred at room temperature for 1 h. The mixture was diluted with DCM (70 mL) and saturated aqueous NH Wash with Cl (50 mL), water (50 mL), and saturated aqueous NaCl (10 mL). The organic layer was dried over MgSO4, filtered and evaporated. Purify the residue by distillation (hexane / EtOAc, 1:9 to 9:1 v / v) to obtain compound 6 (334 mg, 81%) was obtained as a white solid.

[0488] Calculated value: [M+H] + (C 43 H 50 N3O 15 ) m / z = 848.3; ESI measurement Value: [M+H] + (C 43 H 50 N3O 15 ) m / z = 848.3.

[0489] (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-((( (9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanal (4-nitrophenoxy)carbonyl)oxy)methyl (Cyclophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4 Preparation of ,5-triyltriacetate (7) Benzyl alcohol 6 (334 mg, 0.39 mmol) in THF (3 mL) and A solution of bis(4-nitrophenyl) carbonate (240 mg, 0.79 mmol) was added to D IPEA (0.1 mL, 0.57 mmol) was added, and the mixture was stirred at room temperature for 24 h. The solvent was removed under vacuum, followed by column chromatography (hexane / EtOAc, 1:9 The residue was purified by HCl / HCl (HCl:HCl 3:100, ... And got it.

[0490] Calculated value: [M+H] + (C 50 H 53 N4O 19 ) m / z = 1013.3; ESI Measured value: [M+H] + (C 50 H 53 N4O 19 ) m / z = 1013.3.

[0491] (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-((( (9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanal 5-((5S,8S,11S,12R)-11-((S)-s ec-butyl)-12-(2-((S)-2-((1R,2R)-3-(((1S,2R )-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2- Methyl-3-oxopropyl)pyrrolidin-1-yl)-2-oxoethyl)-5,8- Diisopropyl-4,10-dimethyl-3,6,9-trioxo-2,13-dioxa- 4,7,10-Triazatetradecyl)phenoxy)-6-(methoxycarbonyl)tetradecyl Preparation of dihydro-2H-pyran-3,4,5-triyl triacetate (9) PNP carbonate 7 (112 mg, 0.11 mmol) in DMF (1.5 mL) A solution of MMAE 8 (130 mg, 0.16 mmol) was added to DIPEA (50 μL, 0 HOAt (15 mg, 0.11 mmol) and HOAt (15 mg, 0.29 mmol) were added. The mixture was stirred at room temperature for 20 h. EtOAc (100 mL) was added to the mixture. The organic layer was washed with saturated aqueous N The solution was then washed with HCl, water, and saturated aqueous NaCl. The mixture was dried using hexane / EtOA column chromatography. The residue was purified by HCl / HCl (1:9 to 10:0 v / v) to give compound 9 (81 mg, 46%). Obtained as a white solid.

[0492] Calculated value: [M+H] + (C 83 H 115 N8O 23 ) m / z = 1591.8; ESI Actual value: [M+H] + (C 83 H 115 N8O 23 ) m / z = 1591.7.

[0493] (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-amino -3-methylbutanamido)propanamido)-5-((5S,8S,11S,12R) -11-((S)-sec-butyl)-12-(2-((S)-2-((1R,2R)- 3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino) -1-Methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-2-oxo (isoethyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo- 2,13-Dioxa-4,7,10-triazatetradecyl)phenoxy)-6-(meth oxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate ( 10) Preparation A solution of MMAE construct 9 (81 mg, 51 μmol) in DMF (3 mL) was added to piperidine. C. (0.1 mL, 1.0 mmol) was slowly added, and the reaction was stirred at room temperature for 30 minutes. Reversed-phase chromatography was performed using acetonitrile-water (0.1% formic acid, 5-100%) as the eluent. The residue was purified by chromatography. Fractions containing the desired compound were pooled and purified under vacuum. The mixture was concentrated at rt to give compound 10 (62 mg, 89%) as a white solid.

[0494] Calculated value: [M+H] + (C 68 H 105 N8O 21 ) m / z = 1369.7; ESI Actual value: [M+H] + (C 68 H 105 N8O 21 ) m / z = 1369.6.

[0495] (2S,3R,4S,5S,6S)-2-(2-((15S,18S)-4-(1-( 3-(2-((2-(((9H-fluoren-9-yl)methoxy)carbonyl)-1, 2-Dimethylhydrazinyl)methyl)-1H-pyrrolo[2,3-b]pyridin-1-yl )propanoyl)piperidin-4-yl)-1-(9H-fluoren-9-yl)-15 -isopropyl-18-methyl-3,13,16-trioxo-2,7,10-triox sa-4,14,17-triazanonadecanamido)-5-((5S,8S,11S,12 R)-11-((S)-sec-butyl)-12-(2-((S)-2-((1R,2R )-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amine (1-Methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-2- Oxoethyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxoethyl so-2,13-dioxa-4,7,10-triazatetradecyl)phenoxy)-6-( (Methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate Preparation of (12) MMAE-amine 10 (32 mg, 24 μmol) and Fmo in DMF (1 mL) A solution of c-AzaHIPS-PNH acid 11 (27 mg, 29 μmol) was added to DIPEA (1 0 μL, 57 μmol) and HATU (11 mg, 29 μmol) were added to the reaction mixture. The mixture was stirred at room temperature for 1 h. The mixture was diluted with DCM (70 mL) and saturated aqueous NH Wash with Cl (50 mL), water (50 mL), and saturated aqueous NaCl (10 mL). The organic layer was dried over MgSO4, filtered and evaporated. Purify the residue by distillation (hexane / EtOAc, 1:9 to 9:1 v / v) to obtain compound 12 (33 mg, 61%) was obtained as a white solid.

[0496] Calculated value: [M+2H] 2+ (C 123 H 164 N 14 O 29 ) m / z = 1150.6; ESI measured value: [M+2H] 2+ (C 123 H 164 N 14 O 29 ) m / z=1151 .0.

[0497] (2S,3S,4S,5R,6S)-6-(5-((5S,8S,11S,12R)- 11-((S)-sec-butyl)-12-(2-((S)-2-((1R,2R)-3 -(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)- 1-Methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-2-oxo Ethyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo-2 ,13-dioxa-4,7,10-triazatetradecyl)-2-((2S,5S)-1 5-((1-(3-(2-((1,2-dimethylhydrazinyl)methyl)-1H-pyrrolo [2,3-b]pyridin-1-yl)propanoyl)piperidin-4-yl)amino)- 5-Isopropyl-2-methyl-4,7-dioxo-10,13-dioxa-3,6-di Azapentadecanamido)phenoxy)-3,4,5-trihydroxytetrahydro-2 Preparation of H-pyran-2-carboxylic acid (13) To a solution of MMAE compound 12 (33 mg, 14 μmol) in THF (1 mL) at 0 °C A solution of LiOH (3 mg) in water (0.25 mL) was added. The mixture was stirred at room temperature for 1 h. The reaction was quenched by the addition of acetic acid (3 μL, 53 μmol). The organic solvent was removed. After that, acetonitrile-water (0.05% TFA, 5-80%) was used as the eluent. The residue was purified by reverse phase chromatography. Fractions containing the desired compound were pooled. and concentrated in vacuo to give compound 13 (13 mg, 52%) as a white solid.

[0498] Calculated value: [M+H] + (C 86 H 135 N 14 O22 ) m / z = 1716.0; ESI Actual measured value: [M+H] + (C 86 H 135 N 14 O 22 ) m / z = 1715.9.

[0499] Synthesis of PNP intermediate 16 The PNP intermediate was synthesized according to Scheme 2 shown below.

[0500] Scheme 2 [ka] [ka]

[0501] (2S,3R,4S,5S,6S)-2-(4-formylphenoxy)-6-(methoxy) (Dicarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (1 4) Preparation Acetobromo-α-D-glucopyranuronate methyl ester in acetonitrile (20 mL) Sterol 1 (1.0 g, 2.5 mmol) and 4-hydroxybenzaldehyde (0.1 To a mixture of AgO (1.4 g, 6.1 mmol) was added AgO (1.5 g, 1.2 mmol). The reaction mixture was stirred in the dark at room temperature for 20 h. The reaction mixture was then concentrated in vacuo and purified by column chromatography. The residue was purified using chromatography (hexane / EtOAc, 1:9 to 9:1 v / v). Compound 14 (0.52 g, 97%) was obtained as a white solid.

[0502] Calculated value: [M+Na] + (C 20 H 22 NaO 11 ) m / z = 461.1; ESI Measured value: [M+Na]+ (C 20 H 22 NaO 11 ) m / z = 460.8.

[0503] (2S,3R,4S,5S,6S)-2-(4-(hydroxymethyl)phenoxy)- 6-(Methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyltriazol- Preparation of acetate (15) Aldehyde in chloroform (1.45 mL) and iPr-OH (0.36 mL) at 0 °C A solution of hydride 14 (0.175 g, 0.40 mmol) was added to silica gel (0.041 g) and and NaBH4 (0.032 g, 0.85 mmol) were added. The mixture was warmed to room temperature and stirred for 2 h. Acetic acid (0.05 mL, 0.87 mmol) was added to the reaction mixture. The reaction mixture was then concentrated in vacuo and purified by column chromatography (D The residue was purified using CM / MeOH, 0.1:9 to 1:9 v / v, to give compound 15 ( 0.113 g, 70%) was obtained as a white solid.

[0504] Calculated value: [M+Na] + (C 20 H 24 NaO 11 ) m / z = 463.1; ESI Measured value: [M+Na] + (C 20 H 24 NaO 11 ) m / z = 462.8.

[0505] (2S,3S,4S,5R,6S)-2-(methoxycarbonyl)-6-(4-((( (4-nitrophenoxy)carbonyl)oxy)methyl)phenoxy)tetrahydro-2 Preparation of H-pyran-3,4,5-triyl triacetate (16) Alcohol 15 (0.05 g, 0.11 m) in anhydrous THF (2.6 mL) under argon mol) was added to a solution of bis(4-nitrophenyl) carbonate (0.076 g, 0.25 m mol), followed by DIPEA (0.06 mL, 0.35 mmol). The mixture was stirred at room temperature for 18 h. The reaction mixture was not complete by LCMS. Additional bis(4 -nitrophenyl) carbonate (0.02 g, 0.07 mmol) and DIPEA ( 0.005 mL, 0.03 mmol) was added and the reaction was stirred for an additional 2 hours. The material was then concentrated under vacuum and purified by column chromatography (hexane / EtOAc, 1:9 The residue was purified using a 1:1 v / v) mixture to give compound 16 (0.064 g, 93%) as a white solid. Obtained as a solid.

[0506] Calculated value: [M+Na] + (C 27 H 27 NNaO 15 ) m / z = 628.1; ESI Actual value: [M+Na] + (C 27 H 27 NNaO 15 ) m / z = 627.7.

[0507] Synthesis of MMAE construct 23 The MMAE construct was synthesized according to Scheme 3 shown below.

[0508] Scheme 3 [ka]

[0509] TIFF2024170512000089.tif204166

[0510] TIFF2024170512000090.tif47167

[0511] 4-((S)-2-((S)-2-((((9H-fluoren-9-yl)methoxy) Carbonyl)amino)-3-phenylpropanamido)-6-(tritylamino)hexa benzyl ((S)-1-(((S)-1-(((3R,4S,5S)-1-( (S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenyl (1-Methoxy-2-methyl-3-oxopropyl)amino Roridin-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)( Methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl- Preparation of 1-oxobutan-2-yl)(methyl)carbamate (18) PNP 17 (0.048 g, 0.047 mmHg) in DMF (1.5 mL) under argon ol) and MMAE 8 (0.05 g, 0.070 mmol) was added to a mixture of DIPEA ( 0.032 mL, 0.18 mmol) and HOAt (0.11 mmol) were added. The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was then purified by reversed-phase column chromatography (M Directly used for purification using HCl / HCl (1:9 to 10:0 v / v). 18 (0.061 g, 82%) was obtained as a white solid.

[0512] Calculated value: [M+H] + (C 96 H 120 N9O 13 ) m / z = 1606.9; ESI Actual value: [M+H] + (C 96 H 120 N9O 13 ) m / z = 1606.7.

[0513] 4-((S)-2-((S)-2-((((9H-fluoren-9-yl)methoxy) Carbonyl)amino)-3-phenylpropanamido)-6-aminohexanamido)be Ndil((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2- ((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropane- 2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidine-1 -yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino (amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan Preparation of tan-2-yl(methyl)carbamate (19) MMAE derivative 18 (0.061 g, 0.038 mmol) in DCM (3.0 mL) To the solution was added trifluoroacetic acid (0.30 mL, 3.9 mmol). The reaction was allowed to stand at room temperature. The reaction mixture was then concentrated in vacuo and purified by reverse phase column chromatography (elution The residue was analyzed using MeCN / H2O (1:9 to 10:0 v / v) containing 0.1% formic acid in the solution. The fraction was purified to give compound 19 (0.028 g, 54%) as a white solid.

[0514] Calculated value: [M+H] + (C 77 H 106 N9O 13 ) m / z = 1364.8; ESI Actual value: [M+H] + (C 77 H 106 N9O 13 ) m / z = 1363.8.

[0515] (2S,3R,4S,5S,6S)-2-(4-((5S,8S)-5-benzyl-8 -((4-((5S,8S,11S,12R)-11-((S)-sec-butyl)-1 2-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy -1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxo Propyl)pyrrolidin-1-yl)-2-oxoethyl)-5,8-diisopropyl-4 ,10-Dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10-tri (azatetradecyl)phenyl)carbamoyl)-1-(9H-fluoren-9-yl)- 3,6,14-trioxo-2,15-dioxa-4,7,13-triazahexadecane -16-yl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran Preparation of 3,4,5-triyltriacetate (20) Amine 19 (0.028 g, 0.020 mmol) in DMF (1 mL) under argon and PNP sugar 16 (0.018 g, 0.030 mmol) in a solution of HOAt (8.3 m g, 0.061 mmol) and DIPEA (0.014 mL, 0.080 mmol). The reaction mixture was stirred at room temperature for 16 h and was directly carried on to the next step without further purification. was used directly.

[0516] (2S,3R,4S,5S,6S)-2-(4-((((S)-5-((S)-2- Amino-3-phenylpropanamide)-6-((4-((5S,8S,11S,12R )-11-((S)-sec-butyl)-12-(2-((S)-2-((1R,2R) -3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino )-1-Methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-2-o (xoethyl)-5,8-diisopropyl-4,10-dimethyl-3,6,9-trioxo -2,13-dioxa-4,7,10-triazatetradecyl)phenyl)amino)-6 -oxohexyl)carbamoyl)oxy)methyl)phenoxy)-6-(methoxycarbamoyl)oxy (bornyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (21) preparation To a solution of 20 in DMF was added piperidine (0.014 mL, 0.14 mmol). The reaction mixture was stirred at room temperature for 1.5 h. The reaction mixture was then purified by reverse phase column chromatography. (MeCN / HO containing 0.1% trifluoroacetic acid in the eluent, 1:9 to 10:0 v Compound 21 (0.021 g, 65%) was obtained by direct purification using HCl (100%). Obtained as a coloured solid.

[0517] Calculated value: [M+H] + (C 83 H 118 N9O 23 ) m / z = 1608.8; ESI Actual value: [M+H] + (C 83 H 118 N9O 23 ) m / z = 1607.6.

[0518] (2S,3R,4S,5S,6S)-2-(4-((15S,18S)-4-(1-( 3-(2-((2-(((9H-fluoren-9-yl)methoxy)carbonyl)-1, 2-Dimethylhydrazinyl)methyl)-1H-pyrrolo[2,3-b]pyridin-1-yl )propanoyl)piperidin-4-yl)-15-benzyl-18-((4-((5S, 8S,11S,12R)-11-((S)-sec-butyl)-12-(2-((S)- 2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropanol (2-methyl-3-oxopropyl)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidine -1-yl)-2-oxoethyl)-5,8-diisopropyl-4,10-dimethyl-3 ,6,9-trioxo-2,13-dioxa-4,7,10-triazatetradecyl) (phenyl)carbamoyl)-1-(9H-fluoren-9-yl)-3,13,16,24 -tetraoxo-2,7,10,25-tetraoxa-4,14,17,23-tetraa (Hexacosan-26-yl)phenoxy)-6-(methoxycarbonyl)tetrahydro Preparation of 2H-pyran-3,4,5-triyl triacetate (22) Amine 21 (0.020 g, 0.012 mmol) in DMF (1 mL) under argon and a solution of acid 11 (0.014 g, 0.015 mmol) with HATU (0.007 g, 0.018 mmol) and DIPEA (0.009 mL, 0.052 mmol) were added. The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was then analyzed by preparative HPLC (0.05% CI 0.057). MeCN / H2O containing 0.05% trifluoroacetic acid, 1:9 to 9.5:0.5 v / v) Compound 22 (0.011 g, 35%) was obtained as a white solid. was obtained as.

[0519] Calculated value: [M+2H] 2+ (C 138 H 177 N 15 O 31 ) m / z = 1270.1; ESI measured value: [M+2H] 2+ (C 138 H 177 N 15 O 31 ) m / z=1269 .7.

[0520] (2S,3S,4S,5R,6S)-6-(4-((9S,12S)-12-benzyl -9-((4-((5S,8S,11S,12R)-11-((S)-sec-butyl) -12-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydro (1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-o 2-oxopropyl)pyrrolidin-1-yl)-2-oxoethyl)-5,8-diisopropyl -4,10-dimethyl-3,6,9-trioxo-2,13-dioxa-4,7,10- Triazatetradecyl)phenyl)carbamoyl)-22-((1-(3-(2-((1 ,2-Dimethylhydrazinyl)methyl)-1H-pyrrolo[2,3-b]pyridin-1-yl Propanoyl)piperidin-4-yl)amino)-3,11,14-trioxo-2 ,17,20-trioxa-4,10,13-triazadocosyl)phenoxy)-3,4 Preparation of ,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (23) of compound 22 (0.011 g, 0.004 mmol) in THF (0.5 mL) on ice To the solution was added 1M LiOH (0.035 mL, 0.035 mmol). The mixture was stirred on ice for 2 h. Additional 1 M LiOH (0.035 mL, 0.035 mmol) was added. The reaction mixture was then concentrated in vacuo and purified by preparative HPLC (solvent). MeCN / HO containing 0.05% trifluoroacetic acid in the exudate, 0:10 to 7.5:2 The residue was purified using HCl (0.5 v / v) to give compound 23 (0.005 g, 58%) as a white solid. Got it as a body.

[0521] Calculated value: [M+2H] 2+ (C 101 H 149 N15 O 24 ) m / z = 978.1; E SI measured value: [M+2H] 2+ (C 101 H 149 N 15 O 24 ) m / z=977.8 .

[0522] Synthesis of intermediate 28 The intermediate was synthesized according to Scheme 4 shown below.

[0523] Scheme 4 [ka] [ka] [ka]

[0524] Preparation of tert-butyl 4-oxopiperidine-1-carboxylate (25) Piperidone 24 (400 mg, 2.6 mmol, 1.0 eq) in CHCN (12 mL) volume), DIPEA (540 μL, 3.1 mmol, 1.2 equiv.), and BocO (8 A solution of DMAP (32 mg, 0.26 mmol, 50 mg, 3.9 mmol, 1.5 equiv.) l, 0.10 equiv.) was added. After stirring for 2 h, the solution was concentrated and reconstituted in DCM. The solution was washed with 0.1 M HCl, the organics were dried over Na2SO4, filtered, and Concentration gave product 25 as a pale yellow solid which was used without further purification (515 mg , 99%).

[0525] Calculated value: [M+H] + (C 10 H 17 NNaO3) m / z = 222.1; ESI measurement Value: [M+H] + C 10 H 17 NNaO3) m / z = 223.0.

[0526] tert-Butyl 4-((2-(2-(3-(tert-butoxy)-3-oxopropanol Preparation of (ethoxy)ethyl)amino)piperidine-1-carboxylate (26) Compound 25 (515 mg, 2.7 mmol, 1.0 equiv.) in DCE (12 mL) and amino-PEG2-t-butyl ester (740 mg, 3.2 mmol, 1.2 equiv.) To this solution was added STAB (1.1 g, 5.4 mmol, 2.0 equiv.). The mixture was stirred overnight. The solution was concentrated and the residue was used without further purification.

[0527] Calculated value: [M+H] + (C 21 H 41 N2O6) m / z = 417.3; ESI measured value :[M+H] + (C 21 H 41 N2O6) m / z = 417.0.

[0528] tert-Butyl 4-((((9H-fluoren-9-yl)methoxy)carbonyl) (2-(2-(3-(tert-butoxy)-3-oxopropoxy)ethoxy)ethyl Preparation of )amino)piperidine-1-carboxylate (27) Compound 26 (1.1 g, 2.7 mmol, 1.0 equiv) in DCM (10 mL) at 0 °C and DIPEA (920 μL, 5.4 mmol, 2.0 equiv.) l (1.4 g, 2.0 equiv.) was added. The mixture was warmed to room temperature and stirred for 2 h. The mixture was washed with saturated NaHCO3. The organics were dried over Na2SO4 and filtered. The mixture was purified by flash chromatography (hexane / EtOAc, 1:9 to 1:1 v Purify the crude product by HCl / v) to give compound A as a clear oil (1.6 g, 98% over 3 steps). I got 27.

[0529] Calculated value: [M+Na] + (C 36 H 50 N2NaO8) m / z = 661.3; ESI Actual value: [M+Na] + ((C 36 H 50 N2NaO8) m / z = 660.9.

[0530] tert-Butyl 1-(9H-fluoren-9-yl)-3-oxo-4-(piperidinyl) (2-phenyl-4-yl)-2,7,10-trioxa-4-azatridecan-13-oate 8) Preparation Compound 27 (1.6 g, 2.7 mmol) and TIPS (60 To a solution of 100 μL of toluene was added TFA (6 mL). After stirring for 1 h, the solution was concentrated and the eluate was and reversed-phase chromatography using acetonitrile-water (0.05% TFA, 10-70%). The product 28 was obtained as a white solid (800 mg, 68%).

[0531] Calculated value: [M+H] + (C 27 H 35 N2O6) m / z = 483.2; ESI measured value :[M+H] + (C 27 H 35 N2O6) m / z = 482.9.

[0532] Synthesis of intermediate 30 The intermediate was synthesized according to Scheme 5 shown below.

[0533] Scheme 5 [ka] [ka]

[0534] 3-{[(2S)-1-{[(1S,2R,5R,6S,16E,18E,20R,2 1S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16 -tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetramethyl Kuro [19.3.1.110,14.03,5] Hexacosa-10,12,14(26) ,16,18-pentaen-6-yl]oxy}-1-oxopropan-2-yl](methyl Preparation of {ethyl)carbamoyl}propanoic acid (29) Maytansine (180 mg, 0.28 mmol, 1.0 equiv) in DCM (8 mL) A solution of succinic anhydride (33 mg, 0.33 mmol, 1.2 equiv.) was added to DIPEA ( After stirring overnight, the solution was concentrated and by rush chromatography (DCM / MeOH, 100:0 to 10:1 v / v). Purification gave the product 29 as a white solid (191 mg, 92%).

[0535] Calculated value: [M+H] + (C 36 H 49 ClNO 12 ) m / z = 750.3; ESI Actual value: [M+H] + (C 36 H 49 ClNO 12 ) m / z = 749.8.

[0536] 2,3,4,5,6-Pentafluorophenyl 3-{[(2S)-1-{[(1S,2 R,5R,6S,16E,18E,20R,21S)-11-chloro-2-hydroxy- 12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4, 24-Dioxa-9,22-diazatetracyclo[19.3.1.110,14.03, 5]Hexacosa-10,12,14(26),16,18-pentaen-6-yl]ox methyl}-1-oxopropan-2-yl](methyl)carbamoyl}propanoate (30 Preparation of Compound 29 (160 mg, 0.21 mmol, 1.5 equiv) in DCM (3 mL) and A solution of 100 mg of fluorophenol (60 mg, 0.33 mmol, 1.3 equiv.) and perfluorophenol (60 mg, 0.33 mmol, 1.3 equiv.) was added to the DIC (35 mg, 0.28 mmol, 1.3 equiv) was added. After stirring overnight, the solution was filtered and Concentration and flash chromatography (DCM / MeOH, 100:0 to 24:1 v / v) to give the product 30 as a white solid (190 mg, 97%).

[0537] Calculated value: [M+H] + (C 42 H 48 ClF5N3O 12 ) m / z = 916.3; ES Measured value of I: [M+H] + (C 42 H 48 ClF5N3O 12 ) m / z = 915.7.

[0538] Synthesis of maytansine construct 39 The maytansine construct was synthesized according to Scheme 6 shown below.

[0539] Scheme 6 [ka]

[0540] TIFF2024170512000097.tif208163

[0541] TIFF2024170512000098.tif148167

[0542] (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-((t ert-Butoxycarbonyl)amino)-3-methylbutanamido)propanamido)- 5-(hydroxymethyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2 Preparation of H-pyran-3,4,5-triyl triacetate (31) Amine 5 (341 mg, 0.65 mmol) and Boc- A solution of valine-OH (169 mg, 0.78 mmol) was diluted with DIPEA (0.35 mL, 2 0.0 mmol) and HATU (380 mg, 1.0 mmol) were added as eluents. Reversed-phase chromatography using acetonitrile-water (0.05% TFA, 5-90%) The residue was purified by HPLC. Fractions containing the desired compound were pooled and lyophilized to give the compound. Product 31 (386 mg, 82%) was obtained as a white solid.

[0543] Calculated value: [M+H] + (C 33 H 48 N3O 15 ) m / z = 726.3; ESI measurement Value: [M+H] + (C 33 H 48 N3O 15 ) m / z=726.1.

[0544] (2S,3R,4S,5S,6S)-2-(2-((S)-2-((S)-2-((t ert-Butoxycarbonyl)amino)-3-methylbutanamido)propanamido)- 5-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenoxy)-6- (Methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate Preparation of ester (32) Compound 31 (81 mg, 0.11 mmol, 1.0 equiv) in THF (1.5 mL) and bis-p-nitrophenol carbonate (140 mg, 0.67 mmol, 6.0 To the solution of 1.0 equiv. (1.0 equiv.) was added DIPEA (120 μL, 0.67 mmol, 6.0 equiv.). After stirring overnight, the solution was concentrated and purified by flash chromatography (DCM / MeOH, 100 The product 32 was purified by HPLC using a HPLC-MS / ... was obtained as.

[0545] Calculated value: [M+Na] + (C 40 H 50 N4NaO 19 ) m / z = 913.3; ESI Actual measured value: [M+Na] + (C 40 H 50 N4NaO 19 ) m / z = 912.7.

[0546] 4-(1-(((4-((S)-2-((S)-2-((tert-butoxycarbonyl 2S,3R,4-trimethyl-3-methylbutanamido)propanamido)-3-( ... S,5S,6S)-3,4,5-triacetoxy-6-(methoxycarbonyl)tetrahydrofuran 2H-pyran-2-yloxybenzyloxycarbonylpiperidine 4-yl)-1-(9H-fluoren-9-yl)-3-oxo-2,7,10-triol Preparation of 4-azatridecanoic acid (33) Compound 32 (140 mg, 0.16 mmol, 1.0 equiv) in DCM (5 mL) and A solution of 28 (110 mg, 0.17 mmol, 1.5 equiv.) in DIPEA (100 μL) After stirring overnight, the solution was concentrated and the eluate was and reversed-phase chromatography using acetonitrile-water (0.05% TFA, 50-80%). The product 33 was obtained as a white solid (110 mg, 53%).

[0547] Calculated value: [M+Na] + (C 61 H 79 N5NaO 22 ) m / z = 1256.5; ES Measured value of I: [M+Na] + (C 61 H 79 N5NaO 22 ) m / z = 1256.7.

[0548] (2S,3S,4S,5R,6S)-6-(2-((S)-2-((S)-2-((t ert-Butoxycarbonyl)amino)-3-methylbutanamido)propanamido)- 5-(((4-((2-(2-(2-carboxyethoxy)ethoxy)ethyl)amino) Piperidine-1-carbonyl)oxy)methyl)phenoxy)-3,4,5-trihydro Preparation of hydroxytetrahydro-2H-pyran-2-carboxylic acid (34) To a solution of compound 33 (110 mg, 0.089 mmol) in THF (2 mL) at 0 °C 1M LiOH (1.4 mL) was added, and the mixture was allowed to warm to room temperature and stirred for 2 h. The mixture was concentrated to remove THF and purified by elution with acetonitrile-water (0.05% TF) Compound 34 was purified by reverse phase chromatography using 1000 kJ / cm²² of HCl (0–45%) to give a white solid (68 mg, 87%)

[0549] Calculated value: [M+H]+ (C 39 H 62 N5O 17 ) m / z = 872.4; ESI measurement Value: [M+H] + (C 39 H 62 N5O 17 ) m / z = 871.8.

[0550] (2S,3S,4S,5R,6S)-6-{2-[(2S)-2-[(2S)-2-{ [(tert-butoxy)carbonyl]amino}-3-methylbutanamido]propanamido mido]-5-{[4-(N-{2-[2-(2-carboxyethoxy)ethoxy]ethyl }-3-{[(2S)-1-{[(1S,2R,5R,6S,16E,18E,20R, 21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,1 6-Tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetramethyl Cyclo[19.3.1.110,14.03,5]hexacosa-10,12,14(26 ), 16,18-pentaen-6-yl]oxy}-1-oxopropan-2-yl]( Methyl)carbamoyl}propanamido)piperidine-1-carbonyloxy]methyl} Preparation of {phenoxy}-3,4,5-trihydroxyoxane-2-carboxylic acid (35) Compound 34 (18 mg, 0.021 mmol, 1.0 equiv) in DMF (0.6 mL) and 30 (29 mg, 0.033 mmol, 1.5 equiv.) in DIPEA (30 μL). After stirring overnight, the solution was concentrated and the eluate was Then, reversed-phase chromatography was performed using acetonitrile-water (0.05% TFA, 10-95%). The white solid product 35 was obtained as a mixture with compound 29 (3: 2 of compound 29 to compound 35, 20 mg, 36% yield).

[0551] Calculated value: [M+Na] + (C 75 H 107 ClN5NaO 28 ) m / z=1625.7 ;ESI measured value: [M+Na] + (C 75 H 107 ClN5NaO 28 ) m / z=16 24.6.

[0552] (2S,3S,4S,5R,6S)-6-{2-[(2S)-2-[(2S)-2-a 4-(N-(2-methyl-3-methylbutana...

Claims

1. antibody, Drugs, as well as The antibody is linked to the drug and a first cleavable moiety and the first cleavable Cleavable linkers that contain a second cleavable moiety that prevents cleavage of the moiety. A conjugate comprising:

2. The first cleavable moiety is an enzymatically cleavable moiety and the second cleavable moiety is an enzymatically cleavable moiety.

2. The conjugate of claim 1, wherein the functional moiety is a chemically cleavable moiety.

3. The first cleavable moiety is a chemically cleavable moiety and the second cleavable moiety is a 2. The conjugate of claim 1, wherein the moiety is an enzymatically cleavable moiety.

4. The first cleavable moiety is a first enzymatically cleavable moiety and the second cleavable moiety is 2. The conjugate of claim 1, wherein the cleavable moiety is a second enzymatically cleavable moiety. 。

5. The first enzymatically cleavable moiety comprises a first peptide, and the second enzymatically cleavable moiety comprises a The conjugate of claim 4 , wherein the cleavable moiety comprises a second peptide.

6. The first enzymatically cleavable moiety comprises a peptide and the second enzymatically cleavable moiety comprises a peptide. The conjugate of claim 4 , wherein the glycosylating moiety comprises a glycoside.

7. The conjugate has formula (I): 【Chemistry 1】 is a conjugate of Z is CR ４ or N, R １ is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, Alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, Substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl and substituted heterocyclyl; R ２ and R ３ is hydrogen, alkyl, substituted alkyl, alkenyl, substituted a Alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, a Amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy , acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl aryl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heptyl aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; is R ２ and R ３ may be cyclically linked to form a 5- or 6-membered heterocyclyl. Ku, Each R ４ is hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, Amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxyl acylamino, aminoacyl, alkylamido, substituted alkylamido, sulfo aryl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heptyl Heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl , heterocyclyl, and substituted heterocyclyl; Each R ５ is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl , alkynyl, and substituted alkynyl; Each R ６ is an alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, k is an integer from 1 to 10; R ７ is hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted a Alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, a Amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy , acylamino, aminoacyl, alkylamido, substituted alkylamido, aryl , substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, Selected from substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl And, L １ is the first linker, L ２ is a second linker, W １ is the drug, and W ２ is the antibody, L １ , R ６ Or R ７ one of which comprises the second cleavable portion; The conjugate of claim 1.

8. k is 2, and The conjugate has formula (Ia): 【Chemistry 2】 is a conjugate of R ６’ Or R ６’’ comprises the second cleavable moiety, and R ６’ oh Yobi R ６’’ The other of the two is an alkyl, substituted alkyl, alkenyl, substituted Alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heptyl aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; The conjugate of claim 7.

9. The second cleavable moiety is a glycoside-containing enzymatically cleavable moiety. Item 9. The conjugate according to item 8.

10. Formula (Ib): 【Chemistry 3】 The conjugate of claim 9 ,

11. Formula (Ic): 【Chemistry 4】 The conjugate of claim 9 ,

12. k is 2, and The conjugate has formula (Id): 【Chemistry 5】 is a conjugate of R ６’ and R ６’’ is an alkyl, substituted alkyl, alkenyl, substituted a Alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, hetero Aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heptaryl independently selected from tetracyclyl, tetracyclyl, and substituted heterocyclyl; R ７ comprises the second cleavable portion, The conjugate of claim 7.

13. The second cleavable moiety is a glycoside-containing enzymatically cleavable moiety. Item 13. The conjugate according to item 12.

14. Formula (Ie): 【Chemistry 6】 The conjugate of claim 13,

15. k is 2, and The conjugate has the formula (If): 【Chemistry 7】 is a conjugate of R ６’ and R ６’’ is an alkyl, substituted alkyl, alkenyl, substituted a Alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, hetero Aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heptaryl independently selected from tetracyclyl, tetracyclyl, and substituted heterocyclyl; R ８ comprises the second cleavable portion, The conjugate of claim 7.

16. Formula (Ig): 【Chemistry 8】 The conjugate of claim 14,

17. L １ but, -(T １ -V １ ) ａ -(T ２ -V ２ ) ｂ -(T ３ -V ３ ) ｃ -(T ４ -V ４ ) ｄ - wherein a, b, c and d each independently represent 0 or 1; T １ , T ２ , T ３ and T ４ is a covalent bond, (C １ ~C １２ ) alkyl, substituted ( C １ ~C １２ ) alkyl, aryl, substituted aryl, heteroaryl, substituted Heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA) ｗ , (PEG) ｎ , (AA) ｐ , -(CR １３ O H) ｍ -, 4-amino-piperidine (4AP), acetal group, hydrazine, disulfide and esters, each independently selected from the group consisting of ethylenediamine and esters, and EDA is an ethylenediamine moiety. where PEG is polyethylene glycol and AA is an amino acid residue, and each w is 1. to 20, each n is an integer from 1 to 30, each p is an integer from 1 to 20, and each m is an integer from 1 to 12; V １ , V ２ , V ３ and V ４ is a covalent bond, -CO-, -NR １５ --, --NR １５ (C H ２ ) ｑ -、-N １５ (C ６ H ４ )-、-CONR １５ -、-N １５ --、-C( O)O-、-OC(O)-、-O-、-S-、-S(O)-、-SO ２ -、-SO ２ NR １５ --, --NR １５ SO ２ - and -P(O)OH-, each q is an integer from 1 to 6; Each R １３ is hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. are selected independently from the rule, and Each R １５ is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl , aryl, substituted aryl, heteroaryl, substituted heteroaryl, cyclo Alkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl are independently selected from the The conjugate according to any one of claims 7 to 16.

18. L ２ but, -(T ５ -V ５ ) ｅ -(T ６ -V ６ ) ｆ -(T ７ -V ７ ) ｇ -(T ８ -V ８ ) ｈ - wherein e, f, g and h each independently represent 0 or 1; T ５ , T ６ , T ７ and T ８ is a covalent bond, (C １ ~C １２ ) alkyl, substituted ( C １ ~C １２ ) alkyl, aryl, substituted aryl, heteroaryl, substituted Heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA) ｗ , (PEG) ｎ , (AA) ｐ , -(CR １３ O H) ｍ -, 4-amino-piperidine (4AP), acetal group, hydrazine, disulfide and esters, each independently selected from the group consisting of ethylenediamine and esters, and EDA is an ethylenediamine moiety. where PEG is polyethylene glycol and AA is an amino acid residue, and each w is 1. to 20, each n is an integer from 1 to 30, each p is an integer from 1 to 20, and each m is an integer from 1 to 12; V ５ , V ６ , V ７ and V ８ is a covalent bond, -CO-, -NR １５ --, --NR １５ (C H ２ ) ｑ -、-N １５ (C ６ H ４ )-、-CONR １５ -、-N １５ --、-C( O)O-、-OC(O)-、-O-、-S-、-S(O)-、-SO ２ -、-SO ２ NR １５ --, --NR １５ SO ２ - and -P(O)OH-, each q is an integer from 1 to 6; Each R １３ is hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. are selected independently from the rule, and Each R １５ is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl , aryl, substituted aryl, heteroaryl, substituted heteroaryl, cyclo Alkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl are independently selected from the The conjugate according to any one of claims 7 to 17.

19. T １ However, (C １ ~C １２ ) alkyl and substituted (C １ ~C １２ ) alkyl Selected, T ２ , T ３ , and T ４ is an aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and and substituted heterocyclyl, (EDA) ｗ , (PEG) ｎ , (C １ ~C １２ ) Alki substituted (C １ ~C １２ ) alkyl, (AA) ｐ , -(CR １３ O.H. ｍ -, 4- Amino-piperidine (4AP), acetal group, hydrazine, and ester, respectively. are independently selected, and V １ , V ２ , V ３ and V ４ is a covalent bond, -CO-, -NR １５ --, --NR １５ (C H ２ ) ｑ -、-N １５ (C ６ H ４ )-、-CONR １５ -、-N １５ --、-C( O)O-、-OC(O)-、-O-、-S-、-S(O)-、-SO ２ -、-SO ２ NR １５ --, --NR １５ SO ２ -, and -P(O)OH-, Selected, (PEG) ｎ but 【Chemistry 9】 where n is an integer from 1 to 30; EDA has the following structure: 【Chemistry 10】 where y is an integer from 1 to 6 and r is 0 or 1. and 4-Amino-piperidine (4AP) 【Chemistry 11】 and Each R １２ and R １５ is hydrogen, alkyl, substituted alkyl, polyethyleneglycol any two adjacent aryl moieties independently selected from aryl and substituted aryl; DoR １２ the groups may be cyclically linked to form a piperazinyl ring, and R １３ is selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. Selected from 18. The conjugate of claim 17.

20. T １ But (C １ ~C １２ ) alkyl and V １ is -CO-, T ２ is 4AP, and V ２ is a covalent bond, T ３ (PEG) ｎ and V ３ is -CO-, and d is 0; The conjugate according to any one of claims 17 to 19.

21. T １ But (C １ ~C １２ ) alkyl and V １ is -CO-, T ２ is 4AP, and V ２ is a covalent bond, T ３ (PEG) ｎ and V ３ is -CONH-, and T ４ is aryl or substituted aryl, and V ４ is -CO-; The conjugate according to any one of claims 17 to 19.

22. T ５ is a covalent bond, and V ５ is -CO-, and f, g and h are 0; The conjugate according to any one of claims 18 to 20.

23. The drug is selected from an auristatin, a maytansine, and a duocarmycin. The conjugate according to any one of claims 1 to 22.

24. A compound comprising a cleavable linker for linking an antibody to a drug, said cleavable linker A linker capable of binding a first cleavable moiety and preventing cleavage of said first cleavable moiety. A second cleavable portion A compound comprising:

25. The first cleavable moiety is an enzymatically cleavable moiety and the second cleavable moiety is an enzymatically cleavable moiety.

25. The compound of claim 24, wherein the moiety is a chemically cleavable moiety.

26. The first cleavable moiety is a chemically cleavable moiety and the second cleavable moiety is a 25. The compound of claim 24, wherein the moiety is an enzymatically cleavable moiety.

27. The first cleavable moiety is a first enzymatically cleavable moiety and the second cleavable moiety is 25. The compound of claim 24, wherein the cleavable moiety is a second enzymatically cleavable moiety.

28. The first enzymatically cleavable moiety comprises a first peptide, and the second enzymatically cleavable moiety comprises a 28. The compound of claim 27, wherein the cleavable moiety comprises a second peptide.

29. The first enzymatically cleavable moiety comprises a peptide and the second enzymatically cleavable moiety comprises a peptide.

28. The compound of claim 27, wherein the glycosyl moiety comprises a glycoside.

30. The compound has the formula (II): 【Chemistry 12】 is a compound of the formula Z is CR ４ or N, R ２ and R ３ is hydrogen, alkyl, substituted alkyl, alkenyl, substituted a Alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, a Amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy , acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl aryl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heptyl aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; is R ２ and R ３ may be cyclically linked to form a 5- or 6-membered heterocyclyl. Ku, Each R ４ is hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, Amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxyl acylamino, aminoacyl, alkylamido, substituted alkylamido, sulfo aryl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heptyl Heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl , heterocyclyl, and substituted heterocyclyl; Each R ５ is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl , alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl , substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclo aryl, and substituted heterocyclyl; Each R ６ is an alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkoxy, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, k is an integer from 1 to 10; R ７ is hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted a Alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, a Amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy , acylamino, aminoacyl, alkylamido, substituted alkylamido, aryl , substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, Selected from substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl And, L １ is the first linker, L ２ is a second linker, and W １ is a drug, L １ , R ６ Or R ７ one of which comprises the second cleavable portion; 25. The compound of claim 24.

31. k is 2, and The compound has the formula (IIa): 【Chemistry 13】 is a compound of the formula R ６’ Or R ６’’ comprises the second cleavable moiety, and R ６’ oh Yobi R ６’’ The other of the two is an alkyl, substituted alkyl, alkenyl, substituted Alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heptyl aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; 31. The compound of claim 30.

32. The second cleavable moiety is a glycoside-containing enzymatically cleavable moiety.

32. The compound according to claim 31.

33. Formula (IIb): 【Chemistry 14】 33. The compound of claim 32,

34. Formula (IIc): 【Chemistry 15】 33. The compound of claim 32,

35. k is 2, and The compound has the formula (IId): 【Chemistry 16】 is a compound of the formula R ６’ and R ６’’ is an alkyl, substituted alkyl, alkenyl, substituted a Alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, hetero Aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heptaryl independently selected from tetracyclyl, tetracyclyl, and substituted heterocyclyl; R ７ comprises the second cleavable portion, 31. The compound of claim 30.

36. The second cleavable moiety is a glycoside-containing enzymatically cleavable moiety. Item 36. The compound according to item 35.

37. Formula (IIe): 【Chemistry 17】 37. The compound of claim 36,

38. k is 2, and The compound has the formula (IIf): 【Chemistry 18】 is a compound of the formula R ６’ and R ６’’ is an alkyl, substituted alkyl, alkenyl, substituted a Alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, hetero Aryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heptaryl independently selected from tetracyclyl, tetracyclyl, and substituted heterocyclyl; R ８ comprises the second cleavable portion, 31. The compound of claim 30.

39. Formula (IIg): 【Chemistry 19】 39. The compound of claim 38,

40. L １ but, -(T １ -V １ ) ａ -(T ２ -V ２ ) ｂ -(T ３ -V ３ ) ｃ -(T ４ -V ４ ) ｄ - wherein a, b, c and d each independently represent 0 or 1; T １ , T ２ , T ３ and T ４ is a covalent bond, (C １ ~C １２ ) alkyl, substituted ( C １ ~C １２ ) alkyl, aryl, substituted aryl, heteroaryl, substituted Heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA) ｗ , (PEG) ｎ , (AA) ｐ , -(CR １３ O H) ｍ -, 4-amino-piperidine (4AP), acetal group, hydrazine, disulfide and esters, each independently selected from the group consisting of ethylenediamine and esters, and EDA is an ethylenediamine moiety. where PEG is polyethylene glycol and AA is an amino acid residue, and each w is 1. to 20, each n is an integer from 1 to 30, each p is an integer from 1 to 20, and each m is an integer from 1 to 12; V １ , V ２ , V ３ and V ４ is a covalent bond, -CO-, -NR １５ --, --NR １５ (C H ２ ) ｑ -、-N １５ (C ６ H ４ )-、-CONR １５ -、-N １５ --、-C( O)O-、-OC(O)-、-O-、-S-、-S(O)-、-SO ２ -、-SO ２ NR １５ --, --NR １５ SO ２ - and -P(O)OH-, each q is an integer from 1 to 6; Each R １３ is hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. are selected independently from the rule, and Each R １５ is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl , aryl, substituted aryl, heteroaryl, substituted heteroaryl, cyclo Alkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl are independently selected from the A compound according to any one of claims 30 to 39.

41. L ２ but, -(T ５ -V ５ ) ｅ -(T ６ -V ６ ) ｆ -(T ７ -V ７ ) ｇ -(T ８ -V ８ ) ｈ - wherein e, f, g and h each independently represent 0 or 1; T ５ , T ６ , T ７ and T ８ is a covalent bond, (C １ ~C １２ ) alkyl, substituted ( C １ ~C １２ ) alkyl, aryl, substituted aryl, heteroaryl, substituted Heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA) ｗ , (PEG) ｎ , (AA) ｐ , -(CR １３ O H) ｍ -, 4-amino-piperidine (4AP), acetal group, hydrazine, disulfide and esters, each independently selected from the group consisting of ethylenediamine and esters, and EDA is an ethylenediamine moiety. where PEG is polyethylene glycol and AA is an amino acid residue, and each w is 1. to 20, each n is an integer from 1 to 30, each p is an integer from 1 to 20, and each m is an integer from 1 to 12; V ５ , V ６ , V ７ and V ８ is a covalent bond, -CO-, -NR １５ --, --NR １５ (C H ２ ) ｑ -、-N １５ (C ６ H ４ )-、-CONR １５ -、-N １５ --、-C( O)O-、-OC(O)-、-O-、-S-、-S(O)-、-SO ２ -、-SO ２ NR １５ --, --NR １５ SO ２ - and -P(O)OH-, each q is an integer from 1 to 6; Each R １３ is hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. are selected independently from the rule, and Each R １５ is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl , aryl, substituted aryl, heteroaryl, substituted heteroaryl, cyclo Alkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl are independently selected from the The compound according to any one of claims 30 to 40.

42. T １ However, (C １ ~C １２ ) alkyl and substituted (C １ ~C １２ ) alkyl Selected, T ２ , T ３ , and T ４ is an aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and and substituted heterocyclyl, (EDA) ｗ , (PEG) ｎ , (C １ ~C １２ ) Alki substituted (C １ ~C １２ ) alkyl, (AA) ｐ , -(CR １３ O.H. ｍ -, 4- Amino-piperidine (4AP), acetal group, hydrazine, and ester, respectively. are independently selected, and V １ , V ２ , V ３ and V ４ is a covalent bond, -CO-, -NR １５ --, --NR １５ (C H ２ ) ｑ -、-N １５ (C ６ H ４ )-、-CONR １５ -、-N １５ --、-C( O)O-、-OC(O)-、-O-、-S-、-S(O)-、-SO ２ -、-SO ２ NR １５ --, --NR １５ SO ２ -, and -P(O)OH-, Selected, (PEG) ｎ but 【Chemistry 20】 where n is an integer from 1 to 30; EDA has the following structure: 【Chemistry 21】 where y is an integer from 1 to 6 and r is 0 or 1. and 4-Amino-piperidine (4AP) 【Chemical 22】 and Each R １２ and R １５ is hydrogen, alkyl, substituted alkyl, polyethyleneglycol any two adjacent aryl moieties independently selected from aryl and substituted aryl; DoR １２ the groups may be cyclically linked to form a piperazinyl ring, and R １３ is selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. Selected from 41. The compound of claim 40.

43. T １ But (C １ ~C １２ ) alkyl and V １ is -CO-, T ２ is 4AP, and V ２ is a covalent bond, T ３ (PEG) ｎ and V ３ is -CO-, and d is 0; The compound according to any one of claims 40 to 42.

44. T １ But (C １ ~C １２ ) alkyl and V １ is -CO-, T ２ is 4AP, and V ２ is a covalent bond, T ３ (PEG) ｎ and V ３ is -CONH-, and T ４ is aryl or substituted aryl, and V ４ is -CO-; The compound according to any one of claims 41 to 43.

45. T ５ is a covalent bond, and V ５ is -CO-, and f, g and h are 0; The compound according to any one of claims 41 to 43.

46. The drug is selected from an auristatin, a maytansine, and a duocarmycin. The compound according to any one of claims 24 to 45.

47. A conjugate according to any one of claims 1 to 23, and Pharmaceutically acceptable excipients 13. A pharmaceutical composition comprising:

48. Administering to a subject an effective amount of a conjugate according to any one of claims 1 to 23. A method comprising:

49. 1. A method of treating cancer in a subject, comprising: The subject is administered a therapeutically effective amount of a conjugate according to any one of claims 1 to 23. and administering to the subject a pharmaceutical composition comprising the compound of claim 1, wherein said administering treats cancer in said subject. A method that is effective for

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