Activatable tyrosine-protein kinase membrane receptor (ROR) antibody-drug conjugate and its use

Abstract

This disclosure provides masked anti-tyrosine-protein kinase membrane receptor antibody-drug conjugate structures. The antibody-drug conjugate structures include branched linkers, with two or more payloads bound to the antibody for each branched linker. In addition, this disclosure also encompasses compounds and methods for producing such conjugates. In addition, this disclosure also encompasses methods for using the conjugates. In some embodiments, such ROR-ADCs bind to the same human ROR epitope as the antibody containing the heavy chain variable region (VH) and light chain variable region (VL) described herein.

Description

[Technical Field]

[0001] 1. Cross-reference of related applications This application claims the interests of U.S. Provisional Patent Application No. 63 / 514,783, filed on 20 July 2023, the disclosure of which is incorporated herein by reference in its entirety.

[0002] 2. Sequence Listing This application includes an electronic sequence listing file submitted with this application in XML format, the entire contents of which are incorporated herein by reference. The sequence listing XML file submitted with this application is named "14529-147-228_SEQ_LISTING.xml", was created on 17 July 2024, and has a size of 198,709 bytes.

[0003] 3. Field This disclosure generally relates to activatable antibody-drug conjugates (ADCs) that bind to tyrosine protein kinase membrane receptors (RORs, e.g., human RORs) and methods of using them. [Background technology]

[0004] 4.Background Various tumors may exhibit cell surface expression of tyrosine-protein kinase transmembrane receptor (ROR) antigens, as described in more detail in Gentile, et al. (Cancer Res; 71(8) April 15, 2011), Rebagay, et al. (Front. Oncol., 18 April 2012), Zhang, et al. (American Journal of Pathology, Vol. 181, No. 6, December 2012), Henry, et al. (Oncotarget, Vol. 6, No. 37 2015), Zhang, et al. (PLoS ONE 7(3):e31127.), and Bainbridge, et al. (PLoS ONE 9(7):e102695.) (each of which is incorporated herein by reference in its entirety). In addition, ROR expression may not be present or may be limited in normal, for example, non-cancerous tissues, as described in Balakrishnan et al. (Clin Cancer Res. 2017 Jun 15;23(12):3061-3071) (the whole of which is incorporated herein). Therefore, ROR antigens can be used as tumor-specific markers in certain tumors. Examples of tumors and cancers in which ROR expression has been demonstrated include, but are not limited to, pancreatic cancer, ovarian cancer, breast cancer, lung cancer, gastric cancer, melanoma, Ewing's sarcoma, chronic lymphocytic leukemia, mantle cell lymphoma, and B-ALL, as described in Gohil et al. (Oncoimmunology. 2017;6(7):e1326437) (the whole of which is incorporated herein). Other cancers include, but are not limited to, hematological cancers, prostate cancer, colon cancer, kidney cancer, and uterine cancer.

[0005] Activatable antibodies and activatable antibody-drug conjugates (ADCs) are capable of binding to their targets only in specific environments (e.g., in a protease-rich tumor microenvironment), and are therefore useful for precise / environment-dependent target binding. However, the process of developing activatable antibodies and ADCs is slow, labor-intensive, and costly. Therefore, there is a need for activatable ROR antibodies and ADCs. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Gentile, et al., Cancer Res;71(8)April 15,2011 [Non-Patent Document 2] Rebagay,et al.,Front.Oncol.,18 April 2012 [Non-Patent Document 3] Zhang,et al.,American Journal of Pathology,Vol.181,No.6,December 2012 [Non-Patent Document 4] Henry,et al.,Oncotarget,Vol.6,No.37 2015 [Non-Patent Document 5] Zhang, et al., PLoS ONE 7(3):e31127. [Non-Patent Document 6] Bainbridge, et al., PLoS ONE 9(7):e102695. [Non-Patent Document 7] Balakrishnan et al., Clin Cancer Res.2017 Jun 15;23(12):3061-3071 [Non-Patent Document 8] Gohil et al., Oncoimmunology.2017;6(7):e1326437. [Overview of the Initiative]

[0007] 5. Overview This disclosure provides an ADC comprising an antibody that binds to a tyrosine-protein kinase membrane receptor ("ROR-ADC"). In some embodiments, such a ROR-ADC binds to the same human ROR epitope as an antibody comprising the heavy chain variable region (VH) and light chain variable region (VL) described herein.

[0008] Furthermore, this disclosure provides an ADC comprising an activatable antibody that binds a tyrosine-protein kinase membrane receptor ("activatable ROR-ADC"). In some embodiments, such an activatable ROR-ADC binds to the same human ROR epitope as the antibody comprising the heavy chain variable region (VH) and light chain variable region (VL) described herein.

[0009] This disclosure also provides pharmaceutical compositions comprising ROR-ADCs (e.g., activatable ROR-ADCs) comprising an activatable antibody or fragment thereof that binds to ROR (e.g., "activatable ROR antibody") and a drug conjugated thereto (directly or indirectly). Such pharmaceutical compositions include, in some embodiments, activatable ROR-ADCs comprising an antibody or fragment thereof that binds to essentially the same human ROR epitopes as antibodies comprising VH and VL described herein.

[0010] This disclosure also provides methods for treating, preventing, or mitigating ROR-mediated diseases, disorders, or conditions with ROR-ADCs disclosed herein, such as activatable ROR-ADCs, for example, methods for mitigating one or more symptoms of a ROR-mediated disease, disorder, or condition.

[0011] More specifically, the Disclosure provides (a) a ROR antibody, e.g., an activatable ROR antibody, and (b) one or more pyridazine-pyrrolo coupling moieties, e.g., activatable ROR-ADCs, comprising a drug conjugated to a pyridazine-pyrrolo coupling moiety via a linker using, e.g., a hydrazino-iso-pictet-spengler (HIPS) conjugation method.

[0012] Conventionally, HIPS conjugation methods have been used to generate conjugates having one payload per HIPS moiety per aldehyde tag, thereby generating antibody conjugates with a DAR value of up to 4. In some embodiments, ROR-ADCs disclosed herein, e.g., activatable ROR-ADCs, include a branched HIPS linker having two (or more) identical or different payload molecules per HIPS moiety, and thus it is possible to conjugate two (or more) small molecule payloads per aldehyde group in the protein in a single conjugation step. As a result, the use of such branched linkers enables the generation of higher DAR site-specific conjugates (e.g., DAR up to 8) with controlled payload arrangement, which in the context of therapeutic ADCs would result in a greater amount of drug delivered to the target tissue.

[0013] This disclosure provides ROR-ADCs, such as activatable ROR-ADC structures, comprising (a) a ROR antibody that binds to a tyrosine protein kinase membrane receptor (ROR), such as an activatable antibody, (b) a branched HIPS linker, and (c) a drug. This disclosure also includes compounds and methods for generating such conjugates, as well as methods for using the conjugates.

[0014] Aspects of the present disclosure include (a) an activatable ROR antibody; and (b) an activatable ROR-ADC comprising one or more pyridazine-pyrrolo coupling moieties containing one or more drugs conjugated to a pyridazine-pyrrolo coupling moiety via one or more linkers.

[0015] Also provided is an activated activatable ROR-ADC. In some embodiments, the ROR-ADC comprises (a) an activated activatable ROR antibody and (b) one or more pyridazine-pyrrolo coupling moieties conjugated to a drug via a linker, for example, using a hydrazino-iso-pictet-spengler (HIPS) conjugation method. In some embodiments, the ROR-ADC comprises (a) an activated activatable antibody that binds to a tyrosine-protein kinase membrane receptor (ROR), (b) a branched HIPS linker, and (c) a drug. In some embodiments, the ROR-ADC comprises (a) an activated activatable ROR antibody; and (b) one or more pyridazine-pyrrolo coupling moieties conjugated to one or more drugs via one or more linkers.

[0016] In some embodiments, the ROR-ADC (e.g., an activatable ROR-ADC) is represented by formula (I), and the ROR-ADC a. an antibody that binds to a tyrosine-protein kinase membrane receptor (ROR) (e.g., an activatable antibody that binds to ROR); and b. two or more drugs conjugated to a pyridazine-pyrrolo coupling moiety via a linker, respectively comprising

Chemical formula

[0017] In some embodiments, the activatable ROR-ADC is represented by formula (I), and the activatable ROR-ADC is a. Activatable antibodies that bind to tyrosine protein kinase membrane receptors (RORs); and b. Two or more drugs conjugated via a linker to the pyridazine-pyrrolo coupling portion. Includes, [ka] Ab represents an activatable antibody that binds to ROR; Z 1 , Z 2 , and Z 4 Each of them operates independently, CR 4 and; Z 3 CL B -W 2 and; R 1 , R 2 , R 3 , and R 4 These are selected from hydrogen and alkyl, respectively; L A teeth, -(T 1 -V 1 ) a -(T 2 -V 2 ) b -(T 3 -V 3 ) c -(T 4 -V 4 ) d -(T 5 -V 5 ) e -(T 6 -V 6 ) f - It is the first linker, a, b, c, d, e, and f are each independently either 0 or 1, provided that at least one of a, b, c, d, e, and f is 1; T 1 , T 2 , T 3 , T 4 , T 5 and T 6 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 OH) x -, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxyphenyl (PHP), acetal group, hydrazine, disulfide, and ester, each independently selected from -, 4-amino-piperidine (4AP), meta-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxyphenyl (PHP), acetal group, hydrazine, disulfide, and ester, where EDA is the ethylenediamine moiety, PEG is polyethylene glycol, AA is an amino acid residue or amino acid analog, each w is an integer from 1 to 20, each n is an integer from 1 to 30, each p is an integer from 1 to 20, and each x is an integer from 1 to 12; V 1 , V 2 , V 3 , V 4 , V 5 , and V 6 is a covalent bond, -CO-, -NR 15 -, -NR 15 (CH2) q -, -NR 15 (C6H4)-, -CONR 15 -, -NR 15CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR 15 -, -NR 15 The groups SO2- and -P(O)OH- are independently selected, and each q is an integer from 1 to 6; Each R 13 This is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl; Each R 15 This is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; L B teeth, -(T 7 -V 7 ) g -(T 8 -V 8 ) h -(T 9 -V 9 ) i -(T 10 -V 10 ) j -(T 11 -V 11 ) k -(T 12 -V 12 ) l -(T 13 -V 13 ) m - It is a second linker that includes, g, h, i, j, k, l, and m are each independently 0 or 1, provided that at least one of g, h, i, j, k, l, and m is 1; T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 is a covalent bond, (C1-C 12 ) alkyl, substituted (C1-C12 ) alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA) w (PEG) n , (AA) p ,-(CR 13 OH) x -, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxyphenyl (PHP), acetal group, hydrazine, disulfide, and ester, each independently selected from -, 4-amino-piperidine (4AP), meta-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxyphenyl (PHP), acetal group, hydrazine, disulfide, and ester, where EDA is the ethylenediamine moiety, PEG is polyethylene glycol, AA is an amino acid residue or amino acid analog, each w is an integer from 1 to 20, each n is an integer from 1 to 30, each p is an integer from 1 to 20, and each x is an integer from 1 to 12; V 7 , V 8 , V 9 , V 10 , V 11 , V 12 , and V 13 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-, -SO2NR 15 -, -NR 15 The groups SO2- and -P(O)OH- are independently selected, and each q is an integer from 1 to 6; Each R 13 This is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl; Each R 15This is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; s is an integer between 1 and 10; W 1 It is the first drug; W 2 This is the second drug.

[0018] In some embodiments, Z 1 CR 4 That is the case.

[0019] In some embodiments, Z 3 CL B -W 2 That is the case.

[0020] In some embodiments, W 1 and W 2 One or both of these are camptothecin analogs, such as verotecan.

[0021] In some embodiments, L A teeth, -(T 1 -V 1 ) a -(T 2 -V 2 ) b -(T 3 -V 3 ) c -(T 4 -V 4 ) d -(T 5 -V 5 ) e -(T 6 -V 6 ) f - (In the formula, a, b, c, d, e, and f are each independently either 0 or 1, provided that at least one of a, b, c, d, e, and f is 1; T 1 , T 2 , T 3 , T 4 , T 5 and T 6 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 OH) x -, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxyphenyl (PHP), acetal group, hydrazine, disulfide, and ester, each independently selected from -, 4-amino-piperidine (4AP), meta-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxyphenyl (PHP), acetal group, hydrazine, disulfide, and ester, where EDA is the ethylenediamine moiety, PEG is polyethylene glycol, AA is an amino acid residue or amino acid analog, each w is an integer from 1 to 20, each n is an integer from 1 to 30, each p is an integer from 1 to 20, and each x is an integer from 1 to 12; V 1 , V 2 , V 3 , V 4 , V 5 , and V 6 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-, -SO2NR 15 -, -NR 15The groups SO2- and -P(O)OH- are independently selected, and each q is an integer from 1 to 6; Each R 13 This is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl; Each R 15 (This is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.) Includes.

[0022] L A In some embodiments, T 1 (C1-C 12 )alkyl and substituted (C1-C 12 ) Selected from alkyl groups; T 2 , T 3 , T 4 , T 5 and T 6 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 OH) x -, independently selected from 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, acetal group, hydrazine, and ester; V 1 , V 2 , V 3 , V 4 , V 5 , and V 6 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-, -SO2NR 15 -, -NR 15 Independently selected from the group consisting of SO2- and -P(O)OH-; (PEG) n teeth, [ka] And n is an integer from 1 to 30; The EDA has the following structure: [ka] The ethylenediamine moiety has , where y is an integer from 1 to 6, and r is 0 or 1; 4-amino-piperidine (4AP) is, [ka] and; Each R 12 R is independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, and any two adjacent R 12 The groups can be linked in a cyclic manner to form a piperazinyl ring; In further embodiments, a, b, c, and d are each 1; e and f are 0.

[0023] In some embodiments, T 1 , T 2 , T 3 , T 4 , T 5 and T 6 These can be arbitrarily substituted with glycosides.

[0024] In some embodiments, MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP are optionally replaced with glycosides, respectively.

[0025] In some embodiments, the glycoside is selected from glucuronides, galactosides, glucosides, mannosides, fucosides, O-GlcNAc, and O-GalNAc.

[0026] In some embodiments, L A is a linker ( T 1 (C1-C 12 ) is alkyl, V 1 is -CONH-; T 2 is a substitution (C1-C 12 ) is alkyl, V 2 is -CO-; T 3 (AA) p V 3 is non-existent (e.g., covalent bond); T 4 PABC and V 4 is non-existent (e.g., covalent bond); p is an integer between 1 and 10; a, b, c, and d are each 1; (e and f are both 0) That is the case.

[0027] In further embodiments, PABC is substituted with glycosides, for example, the hydrogen atoms of PABC are replaced with glycosides such as glucuronide, galactoside, glucoside, mannoside, fucoside, O-GlcNAc, and O-GalNAc.

[0028] In some embodiments, L B teeth, -(T 7 -V 7 ) g -(T 8 -V 8 ) h-(T 9 -V 9 ) i -(T 10 -V 10 ) j -(T 11 -V 11 ) k -(T 12 -V 12 ) l -(T 13 -V 13 ) m - (In the formula, g, h, i, j, k, l, and m are each independently 0 or 1, provided that at least one of g, h, i, j, k, l, and m is 1; T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 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 OH) x-, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxyphenyl (PHP), acetal group, hydrazine, disulfide, and ester, each independently selected from -, 4-amino-piperidine (4AP), meta-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxyphenyl (PHP), acetal group, hydrazine, disulfide, and ester, where EDA is the ethylenediamine moiety, PEG is polyethylene glycol, AA is an amino acid residue or amino acid analog, each w is an integer from 1 to 20, each n is an integer from 1 to 30, each p is an integer from 1 to 20, and each x is an integer from 1 to 12; V 7 , V 8 , V 9 , V 10 , V 11 , V 12 , and V 13 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-, -SO2NR 15 -, -NR 15 The groups SO2- and -P(O)OH- are independently selected, and each q is an integer from 1 to 6; Each R 13 This is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl; Each R 15 (This is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.) Includes.

[0029] In some embodiments, T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 These can be arbitrarily substituted with glycosides.

[0030] In some embodiments, MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP are optionally replaced with glycosides, respectively.

[0031] In some embodiments, the glycoside is selected from glucuronides, galactosides, glucosides, mannosides, fucosides, O-GlcNAc, and O-GalNAc.

[0032] L B In some embodiments, T 7 It is a covalent bond; T 8 , T 9 , T 10 , T 11 and T 12 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 OH) x -, independently selected from 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, acetal group, hydrazine, and ester; V 7 , V 8 , V 9 , V 10 , V 11 and V 12 is a covalent bond, -CO-, -NR15 -, -NR 15 (CH2) 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-; (PEG) n teeth, [ka] And n is an integer from 1 to 30; The EDA has the following structure: [ka] The ethylenediamine moiety has , where y is an integer from 1 to 6, and r is 0 or 1; 4-amino-piperidine (4AP) is, [ka] and; Each R 12 R is independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, and any two adjacent R 12 The groups can be linked in a cyclic manner to form a piperazinyl ring; Each of g, h, i, j, and k is 1; l and m are both 0.

[0033] In some embodiments, T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , and T12 These can be arbitrarily substituted with glycosides.

[0034] In some embodiments, MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP are optionally replaced with glycosides, respectively.

[0035] In some embodiments, the glycoside is selected from glucuronides, galactosides, glucosides, mannosides, fucosides, O-GlcNAc, and O-GalNAc.

[0036] In some embodiments, L B is a linker ( T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CONH-; T 9 is a substitution (C1-C 12 ) is alkyl, V 9 is -CO-; T 10 (AA) p V 10 is non-existent (e.g., covalent bond); T 11 PABC and V 11 is non-existent (e.g., covalent bond); p is an integer between 1 and 10; Each of g, h, i, j, and k is 1; l and m are both 0.) That is the case.

[0037] In further embodiments, PABC is substituted with glycosides, for example, the hydrogen atoms of PABC are replaced with glycosides such as glucuronide, galactoside, glucoside, mannoside, fucoside, O-GlcNAc, and O-GalNAc.

[0038] In some embodiments, the ROR-ADC (e.g., an activatable ROR-ADC) is given by formula (I): [ka] (In the formula, Ab represents an antibody that binds to ROR (for example, an activatable antibody that binds to ROR); Z 1 , Z 2 , and Z 4 Each of them operates independently, CR 4 and; Z 3 CL B -W 2 and; R 1 , R 2 , R 3 and R 4 is hydrogen and (C1-C 12 ) Selected from alkyl groups; L A The first linker ( T 1 (C1-C 12 ) is alkyl, V 1 is -CONH-; T 2 is a substitution (C1-C 12 ) is alkyl, V 2 is -CO-; T 3 (AA) p (p is an integer from 1 to 20), and V 3 It is a covalent bond; T 4 PABC and V 4 It is a covalent bond; a, b, c, and d are each 1; (e and f are both 0) and; L B The second linker ( T 7 It is a covalent bond, V 7 It is -NHCO-; T 8(C1-C 12 ) is alkyl, V 8 is -CONH-; T 9 is a substitution (C1-C 12 ) is alkyl, V 9 is -CO-; T 10 (AA) p (p is an integer from 1 to 20), and V 10 It is a covalent bond; T 11 PABC and V 11 It is a covalent bond; Each of g, h, i, j, and k is 1; l and m are both 0.) and; s is an integer between 1 and 10; W 1 It is the first drug; W 2 (This is the second drug.) It is represented by [this].

[0039] In some embodiments, the activatable ROR-ADC is given by formula (I): [ka] (In the formula, Ab represents an activatable antibody that binds to ROR; Z 1 , Z 2 , and Z 4 Each of them operates independently, CR 4 and; Z 3 CL B -W 2 and; R 1 , R 2 , R 3 and R 4 is hydrogen and (C1-C 12 ) Selected from alkyl groups; L A The first linker ( T 1(C1-C 12 ) is alkyl, V 1 is -CONH-; T 2 is a substitution (C1-C 12 ) is alkyl, V 2 is -CO-; T 3 (AA) p (p is an integer from 1 to 20), and V 3 It is a covalent bond; T 4 PABC and V 4 It is a covalent bond; a, b, c, and d are each 1; (e and f are both 0) and; L B The second linker ( T 7 It is a covalent bond, V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CONH-; T 9 is a substitution (C1-C 12 ) is alkyl, V 9 is -CO-; T 10 (AA) p (p is an integer from 1 to 20), and V 10 It is a covalent bond; T 11 PABC and V 11 It is a covalent bond; Each of g, h, i, j, and k is 1; l and m are both 0.) and; s is an integer between 1 and 10; W 1 It is the first drug; W 2 (This is the second drug.) It is represented by [this].

[0040] In some embodiments, W 1 and W 2 One or both of these are camptothecin analogs, such as verotecan.

[0041] In some embodiments, the ROR-ADC (e.g., an activatable ROR-ADC) is given by formula (I): [ka] (In the formula, Ab represents an antibody that binds to ROR (for example, an activatable antibody that binds to ROR); Z 1 , Z 2 , and Z 4 Each of them operates independently, CR 4 and; Z 3 CL B -W 2 and; R 1 , R 2 , R 3 and R 4 is hydrogen and (C1-C 12 ) Selected from alkyl groups; L A is a linker ( T 1 It is (C1-C6) alkyl, and V 1 is -CONH-; T 2 is -NHCO(PEG) t (C1-C6) alkylenes substituted with (PEG) t teeth, [ka] And t is an integer from 2 to 10, arbitrarily 8, and V 2 is -CO-; T 3 is (AA)2 and V 3 It is a covalent bond; T 4 This is a PABC substituted with a glycoside, and V4 It is a covalent bond; a, b, c, and d are each 1; (e and f are both 0) and; L B is a linker ( T 7 It is a covalent bond, V 7 It is -NHCO-; T 8 It is (C1-C6) alkyl, and V 8 is -CONH-; T 9 is -NHCO(PEG) t (C1-C6) alkylenes substituted with (PEG) t teeth, [ka] And t is an integer from 2 to 10, arbitrarily 8, and V 9 is -CO-; T 10 is (AA)2 and V 10 It is a covalent bond; T 11 This is a PABC substituted with a glycoside, and V 11 It is a covalent bond; Each of g, h, i, j, and k is 1; l and m are both 0.) and; s is an integer between 1 and 10; W 1 It is the first drug; W 2 (This is the second drug.) It is represented by [this].

[0042] In some embodiments, the activatable ROR-ADC is given by formula (I): [ka] (In the formula, Ab represents an activatable antibody that binds to ROR; Z 1 , Z 2 , and Z 4 Each of them operates independently, CR 4 and; Z 3 CL B -W 2 and; R 1 , R 2 , R 3 and R 4 is hydrogen and (C1-C 12 ) Selected from alkyl groups; L A is a linker ( T 1 It is (C1-C6) alkyl, and V 1 is -CONH-; T 2 is -NHCO(PEG) t (C1-C6) alkylenes substituted with (PEG) t teeth, [ka] And t is an integer from 2 to 10, arbitrarily 8, and V 2 is -CO-; T 3 is (AA)2 and V 3 It is a covalent bond; T 4 This is a PABC substituted with a glycoside, and V 4 It is a covalent bond; a, b, c, and d are each 1; (e and f are both 0) and; L B is a linker ( T 7 It is a covalent bond, V 7 It is -NHCO-; T 8 It is (C1-C6) alkyl, and V 8 is -CONH-; T 9 is -NHCO(PEG)t (C1-C6) alkylenes substituted with (PEG) t teeth, [ka] And t is an integer from 2 to 10, arbitrarily 8, and V 9 is -CO-; T 10 is (AA)2 and V 10 It is a covalent bond; T 11 This is a PABC substituted with a glycoside, and V 11 It is a covalent bond; Each of g, h, i, j, and k is 1; l and m are both 0.) and; s is an integer between 1 and 10; W 1 It is the first drug; W 2 (This is the second drug.) It is represented by [this].

[0043] In some embodiments, T 4 and T 11 One or both of the PABCs are substituted with a glucuronide. In some embodiments, T 1 and T 8 One or both of them are ethyl. In some embodiments, T 2 and T 9 One or both of these are -NHCO(PEG) t It is a C5 alkylene substituted with (PEG) t teeth, [ka] And t is an integer from 5 to 10, arbitrarily 8. In some embodiments, W 1 and W 2 One or both of these are camptothecin analogs, such as verotecan.

[0044] In some embodiments, the ROR-ADC (e.g., an activatable ROR-ADC) is given by formula (II): [ka] (In the formula, Ab represents an antibody that binds to ROR (hereinafter also referred to as a ROR antibody, or, for example, an activatable antibody that binds to ROR); (s is an integer between 1 and 10) It is represented by [this].

[0045] In some embodiments, the activatable ROR-ADC is given by formula (II): [ka] (In the formula, Ab represents an activatable antibody that binds to ROR; (s is an integer between 1 and 10) It is represented by [this].

[0046] In some embodiments, s is an integer between 1 and 8. In some embodiments, s is 4.

[0047] Formula (II) can be prepared by conjugating one or more linker-payloads of Formula (IIa) shown below with a ROR antibody (e.g., an activatable ROR antibody): [ka]

[0048] In some embodiments, a ROR-ADC (e.g., an activatable ROR-ADC) is represented by formula (I) or (II), and a ROR antibody (Ab) (e.g., an activatable ROR antibody) includes an antibody light chain variable (VL) region and an antibody heavy chain variable (VH) region. In some embodiments, the VH region includes VH CDR1, VH CDR2, and VH CDR3 as shown in VH, which includes the amino acid sequence of SEQ ID NO: 25, and the VL region includes VL CDR1, VL CDR2, and VL CDR3 as shown in VL, which includes the amino acid sequence of SEQ ID NO: 26.

[0049] In further embodiments, Ab further comprises a masking peptide. In even further embodiments, the masking peptide can be cleaved, for example, in the tumor microenvironment, thereby activating the ROR-ADC. Thus, Ab is an activatable ROR antibody. In some embodiments, the masking peptide comprises one of the amino acid sequences of SEQ ID NOs: 29, 30, or 31. In some embodiments, the activatable antibody comprises a polypeptide from the N-terminus to the C-terminus that includes the masking peptide and the antibody VL region.

[0050] In some embodiments, the activatable ROR-ADC is represented by formula (I) or (II), and the activatable antibody (Ab) comprises a masking peptide, an antibody light chain variable (VL) region, and an antibody heavy chain variable (VH) region.

[0051] In some embodiments, the masking peptide can be cleaved, for example, in the tumor microenvironment, thereby activating ROR-ADC.

[0052] In further embodiments, the masking peptide comprises one of the amino acid sequences of SEQ ID NOs: 29, 30, or 31. In further embodiments, the VH region comprises VH CDR1, VH CDR2, and VH CDR3 as shown in VH, which comprises the amino acid sequence of SEQ ID NO: 25, and the VL region comprises VL CDR1, VL CDR2, and VL CDR3 as shown in VL, which comprises the amino acid sequence of SEQ ID NO: 26. In some embodiments, the activatable antibody comprises a polypeptide comprising the masking peptide and the antibody VL region from N-terminus to C-terminus.

[0053] In some embodiments, a ROR-ADC (e.g., an activatable ROR-ADC) is represented by formula (I) or (II), and Ab (e.g., an activatable ROR antibody) comprises a VH region including (i) a VH CDR1 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 7, 12, 13, 18, and 27, a VH CDR2 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 8, 14, 19, and 24, and a VH CDR3 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 9, 15, 20, and 28; and (ii) a VL region including a VL CDR1 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 10, 16, and 21, a VL CDR2 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 11, and 22, and a VL CDR3 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 17, and 23.

[0054] In some embodiments, the activatable ROR-ADC is represented by formula (I) or (II), and the activatable antibody (Ab) comprises a VH region including (i) a VH CDR1 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 7, 12, 13, 18, and 27, a VH CDR2 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 8, 14, 19, and 24, and a VH CDR3 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 9, 15, 20, and 28; and (ii) a VL region including a VL CDR1 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 10, 16, and 21, a VL CDR2 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 11, and 22, and a VL CDR3 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 17, and 23.

[0055] In some embodiments, the ROR-ADC is represented by formula (I) or (II), and the antibody (Ab) competes for binding to ROR with either the ROR antibodies disclosed herein or an activatable ROR antibody.

[0056] In some embodiments, the activatable ROR-ADC is represented by formula (I) or (II), and the activatable antibody (Ab) competes with either the ROR antibody disclosed herein or the activatable ROR antibody for binding to ROR.

[0057] In some embodiments, the ROR-ADC (e.g., an activatable ROR-ADC) is represented by formula (I) or (II), where Ab comprises the framework 1 (FR1), framework 2 (FR2), framework 3 (FR3), and / or framework 4 (FR4) sequences, for example, as shown in either one of sequence numbers 25 and 26.

[0058] In some embodiments, the ROR-ADC (e.g., an activatable ROR-ADC) is represented by formula (I) or (II), where Ab comprises a human framework sequence.

[0059] In some embodiments, ROR-ADC is represented by formula (I) or (II), where Ab includes (i) VH containing the amino acid sequence of SEQ ID NO: 25 and VL containing the amino acid sequence of SEQ ID NO: 26.

[0060] In some embodiments, the activatable ROR-ADC is represented by formula (I) or (II), where Ab comprises (i) VH containing the amino acid sequence of SEQ ID NO: 25 and VL containing the amino acid sequence of SEQ ID NO: 26.

[0061] In some embodiments, the activatable ROR-ADC is represented by formula (I) or (II), where Ab comprises a masking peptide, and the masking peptide comprises the amino acid sequence of SEQ ID NO: 29.

[0062] In some embodiments, the activatable ROR-ADC is represented by formula (I) or (II), where Ab comprises a polypeptide, and the polypeptide comprises the amino acid sequence of SEQ ID NO: 34 or 35.

[0063] In further embodiments, Ab further comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 48 or 171.

[0064] In some embodiments, the activatable ROR-ADC is represented by formula (I) or (II), where Ab comprises a masking peptide, and the masking peptide comprises the amino acid sequence of SEQ ID NO: 30.

[0065] In some embodiments, the activatable ROR-ADC is represented by formula (I) or (II), where Ab comprises a polypeptide, and the polypeptide comprises the amino acid sequence of SEQ ID NO: 36 or 37.

[0066] In further embodiments, Ab further comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 48 or 171.

[0067] In some embodiments, the activatable ROR-ADC is represented by formula (I) or (II), where Ab comprises a masking peptide, and the masking peptide comprises the amino acid sequence of SEQ ID NO: 31.

[0068] In some embodiments, the activatable ROR-ADC is represented by formula (I) or (II), where Ab comprises a polypeptide, and the polypeptide comprises the amino acid sequence of SEQ ID NO: 38 or 39.

[0069] In further embodiments, Ab further comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 48 or 171.

[0070] In some embodiments, the antibody Ab is a sequence of formula (VIII) X 1 (fGly')X 2 Z 20 X 3 Z 30 (VIII) (In the formula, 'fGly' is an amino acid residue coupled to the drug via a linker; Z 20 is either a proline (P) or alanine (A) residue; Z 30 is a basic amino acid residue optionally selected from the group consisting of arginine (R), lysine (K), and histidine (H), or an aliphatic amino acid residue optionally selected from the group consisting of alanine (A), glycine (G), leucine (L), valine (V), isoleucine (I), and proline (P); X 1 X may or may not be present, and if present, it may be any amino acid residue, provided that the sequence of formula (VIII) is at the N-terminus of antibody Ab. 1 It exists; X 2 and X 3 (These can be any amino acid residue, independently.) Includes. In a further embodiment, the sequence of formula (VIII) is L(fGly')TPSR (sequence number 146), M(fGly')TPSR (sequence number 147), V(fGly')TPSR (sequence number 148), L(fGly')SPSR (sequence number 149), L(fGly')APSR (sequence number 150), L(fGly')VPSR (sequence number 151), L(fGly')GPSR (sequence number 152), I(fGly')TPAR (sequence number 153), L(fGly')TPSK (sequence number 154), M(fGly')TPSK (sequence number 155), V(fGly')TPSK (sequence number 156), L(fGly The sequence is selected from the group consisting of ')SPSK (sequence number 157), L(fGly')APSK (sequence number 158), L(fGly')VPSK (sequence number 159), L(fGly')GPSK (sequence number 160), L(fGly')TPSA (sequence number 161), I(fGly')TPAA (sequence number 162), M(fGly')TPSA (sequence number 163), V(fGly')TPSA (sequence number 164), L(fGly')SPSA (sequence number 165), L(fGly')APSA (sequence number 166), L(fGly')VPSA (sequence number 167), and L(fGly')GPSA (sequence number 168). In further embodiments, the sequence of formula (VIII) includes L(fGly')TPSR (sequence number 146).

[0071] In some embodiments, Ab is an IgG1 antibody, optionally an IgG1 kappa antibody.

[0072] In some embodiments, the ROR-ADC is represented by formula (I) or (II), where Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 32 or a variant thereof manipulated to be conjugated into a linker payload, or SEQ ID NO: 42 or a variant thereof conjugated into a linker payload, or SEQ ID NO: 48. In further embodiments, Ab comprises one of the peptides SEQ ID NOs: 33, 34, 36, or 38.

[0073] In some embodiments, the ROR-ADC is represented by formula (I) or (II), where Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 169 or a variant thereof manipulated to be conjugated into a linker payload, or SEQ ID NO: 170 or a variant thereof conjugated into a linker payload, or SEQ ID NO: 171. In further embodiments, Ab comprises one of the peptides SEQ ID NOs: 33, 34, 36, or 38.

[0074] In some embodiments, the activatable ROR-ADC is represented by formula (I) or (II), where Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 32 or a variant thereof manipulated to be conjugated into a linker payload, or SEQ ID NO: 42 or a variant thereof conjugated into a linker payload, or SEQ ID NO: 48.

[0075] In further embodiments, Ab comprises one of the peptides SEQ ID NOs: 34, 36, or 38.

[0076] In some embodiments, the activatable ROR-ADC is represented by formula (I) or (II), where Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 169 or a variant thereof manipulated to be conjugated into a linker payload, or SEQ ID NO: 170 or a variant thereof conjugated into a linker payload, or SEQ ID NO: 171. In further embodiments, Ab comprises one of the peptides SEQ ID NOs: 34, 36, or 38.

[0077] In some embodiments, the activatable ROR-ADC is represented by formula (I) or (II), where Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 42 or a variant thereof.

[0078] In further embodiments, Ab comprises one of the peptides SEQ ID NOs: 34, 36, or 38.

[0079] In some embodiments, ROR-ADC is represented by formula (I) or (II), where Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 48. In further embodiments, Ab comprises one of the peptides of SEQ ID NO: 33, 34, 36, or 38.

[0080] In some embodiments, ROR-ADC is represented by formula (I) or (II), where Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 171. In further embodiments, Ab comprises one of the peptides of SEQ ID NO: 33, 34, 36, or 38.

[0081] In some embodiments, the activatable ROR-ADC is represented by formula (I) or (II), where Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 48.

[0082] In further embodiments, Ab comprises one of the peptides SEQ ID NOs: 34, 36, or 38.

[0083] In some embodiments, the activatable ROR-ADC is represented by formula (I) or (II), where Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 171. In further embodiments, Ab comprises one of the peptides SEQ ID NOs: 34, 36, or 38.

[0084] In some embodiments, the activatable ROR-ADC is represented by formula (II), where s is 4 and Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 48 and a polypeptide containing the amino acid sequence of SEQ ID NO: 34. This ROR-ADC is also referred to herein as ADC-11.

[0085] In some embodiments, the activatable ROR-ADC is represented by formula (II), where s is 4 and Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 48 and a polypeptide containing the amino acid sequence of SEQ ID NO: 36. This ROR-ADC is also referred to herein as ADC-13.

[0086] In some embodiments, the activatable ROR-ADC is represented by formula (II), where s is 4 and Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 48 and a polypeptide containing the amino acid sequence of SEQ ID NO: 38. This ROR-ADC is also referred to herein as ADC-M3.

[0087] In some embodiments, the activatable ROR-ADC is represented by formula (II), where s is 4 and Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 171 and a polypeptide containing the amino acid sequence of SEQ ID NO: 34. This ROR-ADC is also referred to herein as ADC-11-sFc.

[0088] In some embodiments, the activatable ROR-ADC is represented by formula (II), where s is 4 and Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 171 and a polypeptide containing the amino acid sequence of SEQ ID NO: 36. This ROR-ADC is also referred to herein as ADC-13-sFc.

[0089] In some embodiments, the activatable ROR-ADC is represented by formula (II), where s is 4 and Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 171 and a polypeptide containing the amino acid sequence of SEQ ID NO: 38. This ROR-ADC is also referred to herein as ADC-M3-sFc.

[0090] In some embodiments, the ROR-ADC is represented by formula (I) or formula (II) disclosed herein, and the ROR-ADC comprises an activatable ROR antibody, the activatable antibody comprising (a) a masking peptide comprising a masking moiety (MM) and a cleavable moiety (CM); and (b) a target binding site (TBM). In further embodiments, the TBM comprises an antibody light chain variable (VL) region and an antibody heavy chain variable (VH) region. In even further embodiments, the VH region comprises VH complementarity determination region 1 (CDR1), VH complementarity determination region 2 (CDR2), and VH complementarity determination region 3 (CDR3) as shown in VH comprising the amino acid sequence of SEQ ID NO: 25, and the VL region comprises VL CDR1, VL CDR2, and VL CDR3 as shown in VL comprising the amino acid sequence of SEQ ID NO: 26. Additionally or alternatively, the activatable antibody is capable of binding to ROR if the CM is cleaved.

[0091] In some embodiments, the ROR-ADC is represented by formula (I) or formula (II) disclosed herein, where Ab binds to both ROR1 and ROR2. In further embodiments, Ab binds to both human ROR1 and human ROR2. Additionally or alternatively, Ab comprises a silent Fc (sFc). In further embodiments, Ab comprises an sFc having the amino acid sequence shown in SEQ ID NO: 44.

[0092] The Disclosure also provides a pharmaceutical composition comprising an activatable ROR-ADC, the activatable ROR-ADC being represented by formula (I) or formula (II), and a pharmaceutically acceptable excipient, wherein the activatable ROR antibody (activatable ROR Ab or activatable Ab) is as described in any embodiment described herein. In some embodiments, such a pharmaceutical composition exhibits a drug-to-antibody ratio (DAR) of the activatable ROR-ADC of about 1 to about 20, for example, about 2 to about 8, about 1 to about 4, about 2 to about 4, about 3 to about 4, about 4, about 1 to about 8, about 2 to about 8, about 3 to about 8, about 4 to about 8, about 5 to about 8, about 6 to about 8, about 6.5 to about 8, about 6 to about 7, about 6.5 to about 7, about 6.5 to about 7.5, about 7 to about 8, about 6.5, about 7, about 7.5, or about 8 DAR.

[0093] This disclosure also provides ROR-ADCs produced by activating an activatable ROR-ADC disclosed herein. Additionally or alternatively, this disclosure also provides ROR-ADCs which are activatable ROR-ADCs disclosed herein, wherein the activatable antibody Ab is activated. In some embodiments, the ADC or Ab is activated by treating an ADC containing an activatable antibody Ab with one or more proteases, where Ab is a masking peptide, and the one or more proteases cleave within the masking peptide. In some embodiments, the one or more proteases include MMP-9. In some embodiments, Ab includes a heavy chain containing the amino acid sequence shown in SEQ ID NO: 48 or 171 and a light chain containing the amino acid sequence shown in SEQ ID NO: 33. In further embodiments, Ab includes a heavy chain containing the amino acid sequence shown in SEQ ID NO: 48 and a light chain containing the amino acid sequence shown in SEQ ID NO: 33.

[0094] In other embodiments, Ab comprises a heavy chain having the amino acid sequence shown in SEQ ID NO: 171 and a light chain having the amino acid sequence shown in SEQ ID NO: 33.

[0095] This disclosure also provides a method for treating cancer or tumors in a subject, comprising administering a ROR-ADC as disclosed herein, for example, an activatable ROR-ADC as disclosed herein, or a pharmaceutical composition as disclosed herein, to the subject. In some embodiments, the ROR-ADC is represented by formula (I) or (II). In further embodiments, the activatable ROR-ADC is represented by formula (I) or (II) or a pharmaceutical composition comprising an activatable ROR-ADC of formula (I) or (II) and a pharmaceutically acceptable excipient, and the activatable ROR antibody is as described in any embodiment herein.

[0096] This specification provides a kit comprising an antibody-drug conjugate or a pharmaceutical composition disclosed herein, and instructions for use. [Brief explanation of the drawing]

[0097] 6. Brief explanation of the drawing [Figure 1] A schematic diagram of an exemplary selection process for self-blocking peptides is shown, as further demonstrated in Example 1.

[0098] [Figure 2A] As further demonstrated in Example 1, exemplary results of the binding of test antibodies to the antigen are provided. The binding of test antibodies (P0 and M1) to ROR1-Fc, as measured by ELISA, is shown. [Figure 2B] As further demonstrated in Example 1, exemplary results of the binding of test antibodies to the antigen are provided. The binding of test antibodies (P0, M2, and M3) to ROR1-Fc, as measured by ELISA, is shown. [Figure 2C] As further demonstrated in Example 1, exemplary results of the binding of test antibodies to the antigen are provided. The binding of test antibodies (P0, M1, M2, and M3) to ROR2-Fc, as measured by ELISA, is shown. [Figure 2D]As further demonstrated in Example 1, exemplary results of the binding of test antibodies to the antigen are provided. The binding of test antibodies (P0, M1, M2, and M3) to HT-29, as measured by FACS, is shown. [Figure 2E] As further demonstrated in Example 1, exemplary results of the binding of test antibodies to the antigen are provided. The binding of test antibodies (P0, M1, M2, and M3) to H226, as measured by FACS, is shown. [Figure 2F] As further demonstrated in Example 1, exemplary results of the binding of test antibodies to the antigen are provided. The binding of test antibodies (P0, M1, M2, and M3) to 293F, as measured by FACS, is shown.

[0099] [Figure 3] A and B provide exemplary differential scanning fluorescence (DSF) results for the test antibodies (M1, M2, and M3), as further shown in Example 2. A plots fluorescence, while B plots d(fluorescence) / dT.

[0100] [Figure 4A] As further demonstrated in Example 2, exemplary capillary electrophoresis results of sodium dodecyl sulfate (CE-SDS) of the test antibody (P0) are provided. [Figure 4B] As further demonstrated in Example 2, exemplary capillary electrophoresis results of sodium dodecyl sulfate (CE-SDS) of the test antibody (M1) are provided. [Figure 4C] As further demonstrated in Example 2, exemplary capillary electrophoresis results of sodium dodecyl sulfate (CE-SDS) of the test antibody (M2) are provided. [Figure 4D] As further demonstrated in Example 2, exemplary capillary electrophoresis results of sodium dodecyl sulfate (CE-SDS) of the test antibody (M3) are provided.

[0101] [Figure 5A]As further demonstrated in Example 2, exemplary capillary isoelectric focusing (cIEF) results of the test antibody (P0) are provided. [Figure 5B] As further demonstrated in Example 2, exemplary capillary isoelectric focusing (cIEF) results of the test antibody (M1) are provided. [Figure 5C] As further demonstrated in Example 2, exemplary capillary isoelectric focusing (cIEF) results of the test antibody (M2) are provided. [Figure 5D] As further demonstrated in Example 2, exemplary capillary isoelectric focusing (cIEF) results of the test antibody (M3) are provided.

[0102] [Figure 6] As further demonstrated in Example 2, exemplary hydrophobic interaction chromatography (HIC) results for the test antibodies (P0, M1, M2, and M3) are provided.

[0103] [Figure 7A] As further demonstrated in Example 2, we provide the results of an exemplary accelerated stability assessment of the test antibody (P0) under incubation at 40°C. [Figure 7B] As further demonstrated in Example 2, we provide the results of an exemplary accelerated stability assessment of the test antibody (M1) under incubation at 40°C. [Figure 7C] As further demonstrated in Example 2, we provide the results of an exemplary accelerated stability assessment of the test antibody (M2) under incubation at 40°C. [Figure 7D] As further demonstrated in Example 2, we provide the results of an exemplary accelerated stability assessment of the test antibody (M3) under incubation at 40°C.

[0104] [Figure 8A] As further demonstrated in Example 2, exemplary accelerated stability evaluation results are provided for the test antibody (P0) after freezing and thawing (3 rounds: FT3; and 6 rounds: FT6). [Figure 8B]As further demonstrated in Example 2, exemplary accelerated stability evaluation results are provided for the test antibody (M1) after freezing and thawing (3 rounds: FT3; and 6 rounds: FT6). [Figure 8C] As further demonstrated in Example 2, exemplary accelerated stability evaluation results are provided for the test antibody (M2) after freezing and thawing (3 rounds: FT3; and 6 rounds: FT6). [Figure 8D] As further demonstrated in Example 2, exemplary accelerated stability evaluation results are provided for the test antibody (M3) after freezing and thawing (3 rounds: FT3; and 6 rounds: FT6).

[0105] [Figure 9A] As further demonstrated in Example 2, the results of an exemplary accelerated stability assessment of the test antibody (P0) at low pH are provided. [Figure 9B] As further demonstrated in Example 2, the results of an exemplary accelerated stability assessment of the test antibody (M1) at low pH are provided. [Figure 9C] As further demonstrated in Example 2, the results of an exemplary accelerated stability assessment of the test antibody (M2) at low pH are provided. [Figure 9D] As further demonstrated in Example 2, the results of an exemplary accelerated stability assessment of the test antibody (M3) at low pH are provided.

[0106] [Figure 10A] As further demonstrated in Example 2, we provide the results of an exemplary accelerated stability assessment of the test antibody (P0) under oxidation. [Figure 10B] As further demonstrated in Example 2, the results of an exemplary accelerated stability assessment of the test antibody (M1) under oxidation are provided. [Figure 10C] As further demonstrated in Example 2, we provide the results of an exemplary accelerated stability assessment of the test antibody (M2) under oxidation. [Figure 10D] As further demonstrated in Example 2, we provide the results of an exemplary accelerated stability assessment of the test antibody (M3) under oxidation.

[0107] [Figure 11A] As further demonstrated in Example 2, exemplary plasma or serum stability results for the test antibody are provided. Plasma stability, indicated by total antibody concentration, is plotted. [Figure 11B] As further demonstrated in Example 2, exemplary plasma or serum stability results for the test antibody are provided. Serum stability, indicated by total antibody concentration, is plotted. [Figure 11C] As further demonstrated in Example 2, exemplary plasma or serum stability results of the test antibody are provided. Stability is plotted against the activated antibody concentration.

[0108] [Figure 12] As further shown in Example 4, we provide exemplary Western blot results showing human IgG from mice injected with 3 mg / kg or 10 mg / kg of P0 or M2.

[0109] [Figure 13] A–C provide exemplary Western blot results showing human IgG from mice injected with 10 mg / kg of M1(A), M2(B), or M3(C), as further shown in Example 4.

[0110] [Figure 14] As further shown in Example 5, exemplary intratumor receptor occupation (RO) results in mice 14 or 96 hours after administration of 10 mg / kg of P0, M1, M2, or M3 are plotted.

[0111] [Figure 15A-1] Exemplary RO flow cytometry plots are shown 14 hours after administration of 10 mg / kg of isotype control, P0, M1, M2, or M3, as further shown in Example 5. Plots for isotype control, P0, and M1 are also shown. [Figure 15A-2]Exemplary RO flow cytometry plots are shown 14 hours after administration of 10 mg / kg of isotype control, P0, M1, M2, or M3, as further shown in Example 5. Plots for isotype control, P0, and M1 are also shown. [Figure 15B-1] Exemplary RO flow cytometry plots are shown 14 hours after administration of a 10 mg / kg isotype control, P0, M1, M2, or M3, as further shown in Example 5. Plots for M2 and M3 are shown. [Figure 15B-2] Exemplary RO flow cytometry plots are shown 14 hours after administration of a 10 mg / kg isotype control, P0, M1, M2, or M3, as further shown in Example 5. Plots for M2 and M3 are shown.

[0112] [Figure 16A-1] Exemplary RO flow cytometry plots are shown 96 hours after administration of 10 mg / kg of isotype control, P0, M1, M2, or M3, as further shown in Example 5. Plots for isotype control, P0, and M1 are also shown. [Figure 16A-2] Exemplary RO flow cytometry plots are shown 96 hours after administration of 10 mg / kg of isotype control, P0, M1, M2, or M3, as further shown in Example 5. Plots for isotype control, P0, and M1 are also shown. [Figure 16B-1] Exemplary RO flow cytometry plots 96 hours after administration of 10 mg / kg isotype control, P0, M1, M2, or M3 are shown, as further demonstrated in Example 5. Plots for M2 and M3 are shown. [Figure 16B-2] Exemplary RO flow cytometry plots 96 hours after administration of 10 mg / kg isotype control, P0, M1, M2, or M3 are shown, as further demonstrated in Example 5. Plots for M2 and M3 are shown.

[0113] [Figure 17] Exemplary IHZ and immunofluorescence (IF) staining results, as further shown in Example 6, are presented. White arrowheads indicate ROR1 staining outside the island, while white arrows indicate ROR1 staining inside the island.

[0114] [Figure 18A] Example 6 further illustrates exemplary IHZ staining results. It shows IHZ staining of ROR on HT-29 and MCF7 cells. [Figure 18B] Exemplary IHZ staining results are shown in Example 6. IHZ staining of ROR and insulin in normal human pancreatic samples #3 and #5 is shown. In the P0+ insulin panel, white arrowheads indicate ROR1 staining in the pancreas, while white arrows indicate ROR1 staining in the inner islets. In the other insulin panels, white arrows indicate pancreatic islets. [Figure 18C] Exemplary IHZ staining results are shown in Example 6. IHZ staining of ROR and insulin in normal human pancreatic samples #6 and #7 is shown. In the P0+ insulin panel, white arrowheads indicate ROR1 staining in the pancreas, while white arrows indicate ROR1 staining in the inner islets. In the other insulin panels, white arrows indicate pancreatic islets. [Figure 18D] Exemplary IHZ staining results are shown in Example 6. IHZ staining of ROR and insulin in normal human pancreatic samples #8 and #9 is shown. In the P0+ insulin panel, white arrowheads indicate ROR1 staining in the pancreas, while white arrows indicate ROR1 staining in the inner islets. In the other insulin panels, white arrows indicate pancreatic islets.

[0115] [Figure 19A]Example 10 provides exemplary in vitro cytotoxicity results of masked and unmasked ROR-ADC+ / -MMP-9 in cell lines, as detailed therein. Results for ADC-00, ADC-11, and ADC-13 with and without MMP-9 activation in hROR1-expressing HEK cells are plotted. [Figure 19B] As detailed in Example 10, we provide exemplary in vitro cytotoxicity results for masked and unmasked ROR-ADC+ / -MMP-9 in cell lines. We plot the results for ADC-13 and ADC-13-sFc with and without MMP-9 activation in HEK cells expressing hROR1. [Figure 19C] As detailed in Example 10, we provide exemplary in vitro cytotoxicity results for masked and unmasked ROR-ADC+ / -MMP-9 in cell lines. We plot the results for ADC-13 and ADC-13-sFc with and without MMP-9 activation in HEK cells expressing hROR2.

[0116] [Figure 20A] Example 11 provides exemplary binding results when ADC- / +MMP-9 activation to ROR1 and ROR2 is measured by ELISA, as detailed therein. Binding of test ADCs (e.g., ADC-00, ADC-11, and ADC-13) to ROR1 without MMP-9 activation is also plotted. [Figure 20B] Example 11 provides exemplary binding results when ADC- / +MMP-9 activation to ROR1 and ROR2 is measured by ELISA, as detailed therein. Binding of test ADCs (e.g., ADC-00, ADC-11, and ADC-13) to ROR1 is plotted using MMP-9 activation. [Figure 20C]Example 11 provides exemplary binding results when ADC- / +MMP-9 activation to ROR1 and ROR2 is measured by ELISA, as detailed therein. Binding of test ADCs (e.g., ADC-00, ADC-11, and ADC-13) to ROR2 without MMP-9 activation is also plotted. [Figure 20D] Example 11 provides exemplary binding results when ADC- / +MMP-9 activation to ROR1 and ROR2 is measured by ELISA, as detailed therein. Binding of test ADCs (e.g., ADC-00, ADC-11, and ADC-13) to ROR2 is plotted using MMP-9 activation.

[0117] [Figure 21A] As detailed in Example 12, exemplary in vivo efficacy data are provided in an MDA-MB-231 triple-negative breast cancer (TNBC) xenograft model, plotting tumor volume. [Figure 21B] As detailed in Example 12, exemplary in vivo efficacy data are provided in an MDA-MB-231 triple-negative breast cancer (TNBC) xenograft model. The percentage of tumor growth inhibition (%TGI) compared to isotype controls at day 29 is shown.

[0118] [Figure 22A] As detailed in Example 13, exemplary in vivo efficacy data are provided in a JEKO-1 mantle cell lymphoma (MCL) xenograft model, plotting tumor volume. [Figure 22B] As detailed in Example 13, exemplary in vivo efficacy data are provided in a JEKO-1 mantle cell lymphoma (MCL) xenograft model, showing the percentage of tumor growth inhibition (%TGI) compared to isotype controls at day 25.

[0119] [Figure 23] Example 15 provides exemplary pharmacokinetic (PK) results in rats, as detailed therein.

[0120] [Figure 24] Example 16 provides exemplary toxicokinetic (TK) results in rats, as detailed therein.

[0121] [Figure 25] This report summarizes the binding, xenograft efficacy, and rat toxicity profiles of ADC-11 and ADC-13.

[0122] [Figure 26] As detailed in Example 14, exemplary in vivo efficacy data are provided in a non-small cell lung cancer (NSCLC) patient-derived xenograft (PDX) model. [Modes for carrying out the invention]

[0123] 7. Detailed explanation This disclosure provides antibody-drug conjugates (ADCs) that bind to ROR (e.g., activatable ADCs) and drugs conjugated thereto (directly or indirectly). Such ROR ADCs (e.g., activatable ROR-ADCs) are useful in compositions and methods for treating, preventing, or mitigating ROR-mediated diseases, disorders, or conditions, which include one or more symptoms of a disease, disorder, or condition. ROR-mediated diseases, disorders, and conditions include a diverse range of cancers, including but not limited to any cancer, in which tumor cells express or overexpress ROR antigens. In addition, activatable ROR-ADCs are useful for killing and / or eliminating tumor cells. The activatable ROR-ADCs described herein are useful in compositions and methods for treating cancer.

[0124] 7.1 Definition The techniques and procedures described or referenced herein include those generally well understood and / or commonly used by those skilled in the art, such as Sambrook et al., Molecular Cloning: A Laboratory Manual (3rd ed. 2001), Current Protocols in Molecular Biology (Ausubel et al. eds., 2003), Therapeutic Monoclonal Antibodies: From Bench to Clinic (An ed. 2009), Monoclonal Antibodies: Methods and Protocols (Albitar ed. 2010), and Antibody Engineering Vols 1 and 2 (Kontermann and Dubel eds., 2nd ed. 2010). Unless otherwise defined herein, technical and scientific terms used herein have meanings generally understood by those skilled in the art. For the purpose of interpreting this Spec., the following definitions of terms shall apply, and where appropriate, the singular form of a term shall also include the plural form, and the plural form of a term shall also include the singular form. In the event of any conflict between any of the definitions of terms provided herein and any of the references incorporated herein by reference, the definitions of terms provided below shall prevail.

[0125] The following terms have the meanings set forth below unless otherwise indicated. Any undefined terms have the meanings recognized in the art.

[0126] "Alkyl" refers to a monovalent saturated aliphatic hydrocarbon group having 1 to 10 carbon atoms, for example, 1 to 6 carbon atoms, or 1 to 5 carbon atoms, or 1 to 4 carbon atoms, or 1 to 3 carbon atoms. This term includes, by example, linear and branched hydrocarbyl groups (e.g., methyl (CH3-), ethyl (CH3CH2-), n-propyl (CH3CH2CH2-), isopropyl ((CH3)2CH-), n-butyl (CH3CH2CH2CH2-), isobutyl ((CH3)2CHCH2-), sec-butyl ((CH3)(CH3CH2)CH-), t-butyl ((CH3)3C-), n-pentyl (CH3CH2CH2CH2CH2-), and neopentyl ((CH3)3CCH2-)).

[0127] The term "substituted alkyl" refers to an alkyl group as defined herein, where one or more carbon atoms in the alkyl chain (excluding the C1 carbon atom) are heteroatoms (e.g., -O-, -N-, -S-, -S(O)). n Optionally substituted with -(n is 0-2), -NR-(R is hydrogen or alkyl), and -NR, -SO-alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-aryl, -SO2-heteroaryl, and -NR a R b Having 1 to 5 substituents selected from the group consisting of R ’ and R ”However, they may be the same or different, and are selected from hydrogen, optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl, and heterocyclic compounds.

[0128] "Alkylene" refers to a divalent aliphatic hydrocarbon group having either a linear or branched chain, preferably 1 to 6 carbon atoms, more preferably 1 to 3 carbon atoms, which may optionally be -O-, -NR 10 -, -NR 10 C(O)-, -C(O)NR 10 - is hindered by one or more groups selected from the following. This term includes, for example, methylene (-CH2-), ethylene (-CH2CH2-), n-propylene (-CH2CH2CH2-), isopropylene (-CH2CH(CH3)-), (-C(CH3)2CH2CH2-), (-C(CH3)2CH2C(O)-), (-C(CH3)2CH2C(O)NH-), (-CH(CH3)CH2-), etc.

[0129] A "substituted alkylene" refers to an alkylene group having 1 to 3 hydrogen atoms that have been replaced by substituents described for carbon in the definition of "substitution" below.

[0130] The term "alkane" refers to alkyl and alkylene groups as defined herein.

[0131] The terms “alkylaminoalkyl”, “alkylaminoalkenyl”, and “alkylaminoalkynyl” refer to an R'NHR”-group, where R' is an alkyl group as defined herein, and R” is an alkylene, alkenylene, or alkynylene group as defined herein.

[0132] The terms "alkaryl" or "aralkyl" refer to alkylene-aryl and substituted alkylene-aryl groups, and alkylene, substituted alkylene, and aryl are defined herein.

[0133] "Alkoxy" refers to an -O-alkyl group, where alkyl is as defined herein. Examples of alkoxys include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, t-butoxy, sec-butoxy, n-pentoxy, and the like. The term "alkoxy" also refers to alkenyl-O-, cycloalkyl-O-, cycloalkenyl-O-, and alkynyl-O-, where alkenyl, cycloalkyl, cycloalkenyl, and alkynyl are as defined herein.

[0134] The term "substituted alkoxy" refers to the groups of substituted alkyl-O-, substituted alkenyl-O-, substituted cycloalkyl-O-, substituted cycloalkenyl-O-, and substituted alkynyl-O-, where substituted alkyl, substituted alkenyl, substituted cycloalkyl, substituted cycloalkenyl, and substituted alkynyl are as defined herein.

[0135] The term "alkoxyamino" refers to an -NH-alkoxy group, where alkoxy is defined herein.

[0136] The term "haloalkoxy" refers to an alkyl-O- group in which one or more hydrogen atoms on an alkyl group are substituted with a halo group, including, for example, trifluoromethoxy groups.

[0137] The term "haloalkyl" refers to a substituted alkyl group as described above, in which one or more hydrogen atoms on the alkyl group are substituted with a halo group. Examples of such groups include, but are not limited to, fluoroalkyl groups (e.g., trifluoromethyl, difluoromethyl, trifluoroethyl, etc.).

[0138] The term "alkylalkoxy" refers to the groups of alkylene-O-alkyl, alkylene-O-substituted alkyl, substituted alkylene-O-alkyl, and substituted alkylene-O-substituted alkyl, where alkyl, substituted alkyl, alkyl, alkylene, and substituted alkylene are as defined herein.

[0139] The term "alkylthioalkoxy" refers to the groups of alkylene-S-alkyl, alkylene-S-substituted alkyl, substituted alkylene-S-alkyl, and substituted alkylene-S-substituted alkyl, where alkyl, substituted alkyl, alkyl, alkylene, and substituted alkylene are as defined herein.

[0140] "Alkenyl" refers to a linear or branched hydrocarbon group having 2 to 6 carbon atoms, preferably 2 to 4 carbon atoms, and at least one, preferably 1 to 2, double bond unsaturated sites. This term includes, as examples, bivinyl, allyl, and buto-3-en-1-yl. This term includes cis isomers and trans isomers, or mixtures thereof.

[0141] The term "substituted alkenyl" refers to an alkenyl group as defined herein having 1 to 5 substitutions or 1 to 3 substituents, which is selected from alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azide, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl, and -SO2-heteroaryl.

[0142] "Alkynyl" refers to a monovalent hydrocarbon group in a linear or branched chain having 2 to 6 carbon atoms, preferably 2 to 3 carbon atoms, and at least 1, preferably 1 to 2 triple bond unsaturated sites. Examples of such alkynyl groups include acetylenyl (-C≡CH) and propargyl (-CH2C≡CH).

[0143] The term "substituted alkynyl" refers to an alkynyl group as defined herein having 1-5 substitutions or 1-3 substituents, which is selected from alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azide, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl, and -SO2-heteroaryl.

[0144] "Alkynyloxy" refers to an -O-alkynyl group, where alkynyl is as defined herein. Examples of alkynyloxy include ethynyloxy and propynyloxy.

[0145] "Acyl" includes HC(O)-, alkyl-C(O)-, substituted alkyl-C(O)-, alkenyl-C(O)-, substituted alkenyl-C(O)-, alkynyl-C(O)-, substituted alkynyl-C(O)-, cycloalkyl-C(O)-, substituted cycloalkyl-C(O)-, cycloalkenyl-C(O)-, substituted cycloalkenyl-C(O)-, aryl-C(O)-, substituted aryl-C(O)-, heteroaryl-C(O)- This refers to substituted heteroaryl-C(O)-, heterocyclyl-C(O)-, and substituted heterocyclyl-C(O)- groups, and alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic are as defined herein. For example, acyl contains the "acetyl" group CH3C(O)-.

[0146] "Acylamino" is -NR 20 C(O)alkyl, -NR 20 C(O) substituted alkyl, NR 20 C(O) cycloalkyl, -NR 20 C(O) substituted cycloalkyl, -NR 20 C(O)cycloalkenyl, -NR 20 C(O)-substituted cycloalkenyl, -NR 20 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) heteroaryl, -NR 20 C(O) substituted heteroaryl, -NR 20 C(O) heterocycle, and -NR 20 It is a C(O) substitution heterogly, and R 20The elements are hydrogen or alkyl, and alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic are as defined herein.

[0147] The terms "aminocarbonyl" or "aminoacyl" are derived from -C(O)NR 21 R 22 It refers to the base, R 21 and R 22 However, independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic, R 21 and R 22 However, optionally, they may bond with the nitrogen atom bonded to them to form heterocyclic or substituted heterocyclic groups, where alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic groups are as defined herein.

[0148] "Aminocarbonylamino" is -NR 21 C(O)NR 22 R 23 It refers to the base, R 21 , R 22 , and R 23 However, independently, the R groups are selected from hydrogen, alkyl, aryl, or cycloalkyl, or two R groups are bonded together to form a heterocycline group.

[0149] The term "alkoxycarbonylamino" refers to the -NRC(O)OR group, where each R is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, or heterocyclyl, with alkyl, substituted alkyl, aryl, heteroaryl, and heterocyclyl being as defined herein.

[0150] The term "acyloxy" refers to the groups alkyl-C(O)O-, substituted alkyl-C(O)O-, cycloalkyl-C(O)O-, substituted cycloalkyl-C(O)O-, aryl-C(O)O-, heteroaryl-C(O)O-, and heterocyclyl-C(O)O-, where alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, heteroaryl, and heterocyclyl are as defined herein.

[0151] "Aminosulfonyl" is -SO2NR 21 R 22 It refers to the base, R 21 and R 22 However, independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic, R 21 and R 22 However, optionally, together with the nitrogen bonded thereto, they may form heterocyclic or substituted heterocyclic groups, and alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic are as defined herein.

[0152] "Sulfonylamino" is -NR 21 SO2R 22 It refers to the base, R 21 and R 22However, independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic, R 21 and R 22 However, optionally, together with the atoms bonded to them, they may form heterocyclic or substituted heterocyclic groups, where alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic are as defined herein.

[0153] "Aryl" or "Ar" refers to a monovalent aromatic carbocyclic group of 6 to 18 carbon atoms having a single ring (e.g., present in a phenyl group) or a ring system having multiple fused rings (examples of such aromatic ring systems include naphthyl, anthryl, and indanyl), where the fused rings may or may not be aromatic, but the bonding sites are mediated by atoms of the aromatic ring. This term includes, by example, phenyl and naphthyl. Unless otherwise limited by the definition of aryl substituents, such aryl groups may be optionally substituted with 1 to 5 substituents or 1 to 3 substituents, which are selected from acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted alkoxy, substituted alkenyl, substituted alkynyl, substituted cycloalkyl, substituted cycloalkenyl, amino, substituted amino, aminoacyl, acylamino, alkaryl, aryloxy, azide, carboxyl, carboxylalkyl, cyano, halogen, nitro, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl, -SO2-heteroaryl, and trihalomethyl.

[0154] "Aryloxy" refers to an -O-aryl group, where aryl is as defined herein, including, for example, phenoxy, naphthoxy, and optionally, optionally substituted aryl groups as defined herein.

[0155] "Amino" refers to the -NH2 group.

[0156] The term "substituted amino" refers to a -NRR group, where each R is independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkynyl, substituted alkynyl, aryl, heteroaryl, and heterocyclyl, provided that at least one R is not hydrogen.

[0157] The term "azide" refers to the -N3 group.

[0158] "Carboxyl," "carboxy," or "carboxylate" refers to -CO2H or a salt thereof.

[0159] The terms "carboxyl ester" or "carboxyester," or "carboxyalkyl" or "carboxyalkyl" refer to -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, and -C(O)O-cycloalkenyl -C(O)O-substituted cycloalkenyl, -C(O)O-heteroaryl, -C(O)O-substituted heteroaryl, -C(O)O-heterocyclic, and -C(O)O-substituted heterocyclic groups are defined herein as follows: alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic groups.

[0160] "(carboxyl ester)oxy" or "carbonate" refers to -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 aryl, -OC(O)O-cycloalkyl, -OC(O)O-substituted cycloalkyl, -OC(O)O-cycloalkenyl, -OC(O)O-substituted cyclo Roalkenyl, -OC(O)O-heteroaryl, -OC(O)O-substituted heteroaryl, -OC(O)O-heterocyclic, and -OC(O)O-substituted heterocyclic are defined herein as alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic are defined herein.

[0161] "Cyano" or "nitrile" refers to the -CN group.

[0162] "Cycloalkyl" refers to a cyclic alkyl group of 3 to 10 carbon atoms having one or more cyclic rings (e.g., fused rings, crosslinked rings, and spiro-ring systems). Suitable examples of cycloalkyl groups include, for example, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclooctyl. Such cycloalkyl groups include, for example, monocyclic structures (e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, etc.) or polycyclic structures (e.g., adamantanyl, etc.).

[0163] The term "substituted cycloalkyl" refers to cycloalkyl groups having 1 to 5 substituents or 1 to 3 substituents, including alkyl, substituted alkyl, alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azide, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO 2- Selected from alkyl, -SO2-substituted alkyl, -SO2-aryl, and -SO2-heteroaryl.

[0164] "Cycloalkenyl" refers to a non-aromatic cyclic alkyl group having 3 to 10 carbon atoms, having a monocyclic or polycyclic structure and at least one double bond, preferably one to two double bonds.

[0165] The term "substituted cycloalkenyl" refers to a cycloalkenyl group having 1 to 5 substituents or 1 to 3 substituents, selected from alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azide, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl, and -SO2-heteroaryl.

[0166] "Cycloalkynyl" refers to a non-aromatic cycloalkyl group consisting of 5 to 10 carbon atoms, having a monocyclic or polycyclic structure and at least one triple bond.

[0167] "Cycloalkoxy" refers to -O-cycloalkyl.

[0168] "Cycloalkenyloxy" refers to -O-cycloalkenyl.

[0169] "Halo" or "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0170] "Hydroxy" or "hydroxyl" refers to the -OH group.

[0171] A "heteroaryl" refers to an aromatic group having 1 to 15 carbon atoms (e.g., 1 to 10 carbon atoms) in a ring and 1 to 10 heteroatoms selected from the group consisting of oxygen, nitrogen, and sulfur. Such heteroaryl groups may have a monocyclic ring (e.g., pyridinyl, imidazolyl, or furyl) or a fused ring system (e.g., groups such as indolidinyl, quinolinyl, benzofuran, benzimidazolyl, or benzothienyl), with at least one ring in the ring system being aromatic. To satisfy the valence requirement, the heteroatoms in such a heteroaryl ring may or may not be bonded to H or substituents, e.g., alkyl groups or other substituents as described herein. In certain embodiments, the nitrogen and / or sulfur ring atoms(s) of the heteroaryl group may be optionally oxidized to provide an N-oxide (N→O), sulfinyl, or sulfonyl moiety. This term includes, by example, pyridinyl, pyrrolyl, indolyl, thiophenyl, and furanyl. Unless otherwise limited by the definition of heteroaryl substituents, such heteroaryl groups may be optionally substituted with 1 to 5 substituents or 1 to 3 substituents, selected from acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted alkoxy, substituted alkenyl, substituted alkynyl, substituted cycloalkyl, substituted cycloalkenyl, amino, substituted amino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, azide, carboxyl, carboxylalkyl, cyano, halogen, nitro, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl, and -SO2-heteroaryl, as well as trihalomethyl.

[0172] The term "heteroaralkyl" refers to an alkylene-heteroaryl group, where alkylene and heteroaryl are defined herein. This term includes, for example, pyridylmethyl, pyridylethyl, indolylmethyl, and others.

[0173] "Heteroaryloxy" refers to -O-heteroaryl.

[0174] "Heterocyclic," "heterocyclic," "heterocycloalkyl," and "heterocyclyl" refer to saturated or unsaturated groups having a monocyclic or multiple fused rings, including fused-bridged ring systems and spirocyclic systems, and having 3 to 20 ring atoms containing 1 to 10 heteroatoms. These ring atoms are selected from nitrogen, sulfur, or oxygen, and in fused ring systems, one or more of the rings may be cycloalkyl, aryl, or heteroaryl, provided that the bonding site is via a non-aromatic ring. In certain embodiments, the nitrogen and / or sulfur atoms of the heterocyclic group are optionally oxidized to provide an N-oxide, -S(O)-, or -SO2- moiety. To satisfy valence requirements, the heteroatoms in such heterocyclic rings may or may not be bonded to one or more H or one or more substituents (e.g., alkyl groups or other substituents as described herein).

[0175] Examples of heterocyclic and heteroaryl compounds include azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indidine, isoindole, indole, dihydroindole, indazole, purine, quinoridine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carborin, phenantholidine, acridine, phenanthroline, isothiazole, phenazine, isoxazoline Examples of such substances include, but are not limited to, phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, indoline, phthalimide, 1,2,3,4-tetrahydroisoquinoline, 4,5,6,7-tetrahydrobenzo[b]thiophene, thiazole, thiazolidine, thiophene, benzo[b]thiophene, morpholinil, thiomorpholinil (also called thiamorpholinil), 1,1-dioxothiomorpholinil, piperidinil, pyrrolidine, and tetrahydrofuranil.

[0176] Unless specifically limited by the definition of heterocyclic substituents, such heterocyclic groups can be optionally substituted with 1 to 5 or 1 to 3 substituents, which include alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azide, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO 2- Selected from alkyl, -SO2-substituted alkyl, -SO2-aryl, -SO2-heteroaryl, and condensed heterocycles.

[0177] "Heterocyclyloxy" refers to the -O-heterocyclyl group.

[0178] The term "heterocyclilthio" refers to a heterocyclic -S- group.

[0179] The term "heterocyclene" refers to a diradical group formed from a heterocycle as defined herein.

[0180] The term "hydroxyamino" refers to the -NHOH group.

[0181] "Nitro" refers to the -NO2 group.

[0182] "Oxo" refers to an atom (=O).

[0183] "Sulfonyl" refers to groups of -SO2-alkyl, -SO2-substituted alkyl, -SO2-alkenyl, -SO2-substituted alkenyl, -SO2-cycloalkyl, -SO2-substituted cycloalkyl, -SO2-cycloalkenyl, -SO2-substituted cycloalkenyl, -SO2-aryl, -SO2-substituted aryl, -SO2-heteroaryl, -SO2-substituted heteroaryl, -SO2-heterocyclic, and -SO2-substituted heterocyclic, where alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic are as defined herein. Examples of sulfonyls include methyl-SO2-, phenyl-SO2-, and 4-methylphenyl-SO2-.

[0184] "Sulfonyloxy" refers to the groups of -OSO2-alkyl, -OSO2-substituted alkyl, -OSO2-alkenyl, -OSO2-substituted alkenyl, -OSO2-cycloalkyl, -OSO2-substituted cycloalkyl, -OSO2-cycloalkenyl, -OSO2-substituted cycloalkenyl, -OSO2-aryl, -OSO2-substituted aryl, -OSO2-heteroaryl, -OSO2-substituted heteroaryl, -OSO2-heterocyclic, and -OSO2-substituted heterocyclic, where alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic are as defined herein.

[0185] "Sulfate" or "sulfate ester" refers to -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-SO2-O-substituted aryl, -O-SO2-O -Heteroaryl, -O-SO2-O-substituted heteroaryl, -O-SO2-O-heterocyclic, and -O-SO2-O-substituted heterocyclic groups, wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic groups are as defined herein.

[0186] The term "aminocarbonyloxy" refers to the -OC(O)NRR group, where each R is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, or heterocyclic, with alkyl, substituted alkyl, aryl, heteroaryl, and heterocyclic being as defined herein.

[0187] "Thiol" refers to the -SH group.

[0188] The terms "thioxo" or "thioketo" refer to an atom (=S).

[0189] The terms "alkylthio" or "thioalkoxy" refer to an -S-alkyl group, where alkyl is as defined herein. In certain embodiments, sulfur may be oxidized to -S(O)-. The sulfoxide may exist as one or more stereoisomers.

[0190] The term "substituted thioalkoxy" refers to an -S-substituted alkyl group.

[0191] The term "thioaryloxy" refers to an aryl-S- group, where the aryl group is as defined herein, and this includes optionally substituted aryl groups as also defined herein.

[0192] The term "thioheteroaryloxy" refers to a heteroaryl-S-group, where the heteroaryl group is as defined herein, and this includes any optionally substituted aryl groups as defined herein.

[0193] The term "thioheterocyclooxy" refers to a heterocyclyl-S-group, where the heterocyclyl group is as defined herein, and this includes any optionally substituted heterocyclyl groups as defined herein.

[0194] In the disclosure herein, the term “substituted” may also mean, when used to modify a particular group or radical, that one or more hydrogen atoms of a particular group or radical are each independently replaced by the same or different substituents as defined below.

[0195] In addition to the groups disclosed with respect to individual terms herein, substituents for substituting one or more hydrogens on a saturated carbon atom or radical of a given group (any two hydrogens on a single carbon, =O, =NR) are also available. 70、=N-OR 70 (which can be replaced with =N2 or =S) Unless otherwise specified, -R 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 , -OSO2R 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 ,-OC(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 70 C(S)R 70 , -NR 70 CO2 - M + , -NR 70 CO2R 70 , -NR 70 C(S)OR 70 , -NR 70C(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 However, selected from the group consisting of optionally substituted alkyl, cycloalkyl, heteroalkyl, heterocycloalkylalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl, each R 70 However, independently, hydrogen or R 60 and; each R 80 However, independently, R 70 or, instead, two R's 80 However, together with the nitrogen atom to which they are bonded, they form a 5, 6, or 7-membered heterocycloalkyl group, which may optionally contain 1 to 4 additional identical or different heteroatoms selected from the group consisting of O, N, and S, of which N may have a -H or C1-C3 alkyl substitution, and each M + However, each M is a counterion with a single positive charge. + However, independently, for example, alkaline ions (for example, K + na + Li + ); ammonium ions (for example, + N(R 60 )4); or alkaline earth ions (e.g., [Ca 2+ ] 0.5 [Mg 2+ ] 0.5 , or [Ba 2+ ] 0.5 ) may be (the subscript 0.5 indicates that one of the counterions of such a divalent alkaline earth ion is the ionized form of the compound of the present invention, and another representative counterion (e.g., chloride), or two ionized compounds disclosed herein, may function as counterions of such a divalent alkaline earth ion, or the biionized compound of the present invention may function as a counterion of such a divalent alkaline earth ion). For example, -NR80 R 80 This means that it includes -NH2, -NH-alkyl, N-pyrrolidinyl, N-piperazinyl, 4N-methylpiperazin-1-yl, and N-morpholinyl.

[0196] In addition to the disclosures herein, the substituents of hydrogen on unsaturated carbon atoms in "substituted" alkenes, alkynes, aryls, and heteroaryl groups are -R unless otherwise specified. 60 Hello, -O - M + , -OR 70 , -SR 70 , -S - M + , -NR 80 R 80 , trihalomethyl, -CF3, -CN, -OCN, -SCN, -NO, -NO2, -N3, -SO2R 70 , -SO3M + , -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 70 , -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 , -OCO2 - M + , -OCO2R 70 , -OC(S)OR 70 , -NR 70 C(O)R70 , -NR 70 C(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 70 )NR 80 R 80 And R 60 , R 70 , R 80 , and M + However, as defined above, except in the case of substituted alkenes or alkynes, the substituent is -O - M + , -OR 70 , -SR 70 , or -S - M + isn't it.

[0197] In addition to the groups disclosed in this specification with respect to individual terms, the substituents on the nitrogen atom of "substituted" heteroalkyl and cycloheteralkyl groups are, unless otherwise specified, -R 60 , -O - M + , -OR 70 , -SR 70 , -S - M + , -NR 80 R 80 , 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 70 )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 + However, it is as defined above.

[0198] In addition to the disclosure herein, in certain embodiments, the substituted group has one, two, three, or four substituents, one, two, or three substituents, one or two substituents, or one substituent.

[0199] It is understood that polymers obtained by substituting the substituents themselves with further substituents (for example, substituted aryl groups having a substituted aryl group as a substituent (which itself is substituted with a substituted aryl group that is further substituted with a substituted aryl group)) are not intended to be included herein. In such cases, the maximum number of substitutions is three. For example, the sequential substitutions of substituted aryl groups that are specifically considered herein are limited to substituted aryl-(substituted aryl)-substituted aryl.

[0200] Unless otherwise specified, substituents not explicitly defined herein are nominated by naming the terminal portion of the functional group, followed by the functional groups adjacent to the bond site. For example, the substituent "arylalkyloxycarbonyl" refers to the (aryl)-(alkyl)-OC(O)- group.

[0201] It is understood that, for any of the groups disclosed herein containing one or more substituents, such groups do not contain any substitutions or substitution patterns that are sterically unrealistic and / or synthetically impossible. In addition, the compounds of the subject matter include all stereochemical isomers resulting from the substitutions of these compounds.

[0202] The term “pharmaceutically acceptable salt” means a salt that is acceptable for administration to a patient (e.g., a mammal) (a salt containing a counterion that has mammalian safety acceptable for a given dosing regimen). Such salts can be derived from pharmaceutically acceptable inorganic or organic bases and pharmaceutically acceptable inorganic or organic acids. “pharmaceutically acceptable salt” refers to a pharmaceutically acceptable salt of a compound, which is derived from a variety of organic and inorganic counterions well known in the art, and includes, but are not limited to, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, etc., and also includes salts of organic or inorganic acids (e.g., hydrochloride, hydrobromide, formate, tartrate, besylate, mesylate, acetate, maleate, oxalate, etc.) where the molecule contains a basic functional group.

[0203] The term "salt" refers to a compound formed when a proton of an acid is replaced by a cation (e.g., a metallic cation or an organic cation). Where applicable, a salt is a pharmaceutically acceptable salt, which is not required for salts of intermediate compounds not intended for administration to a patient. As an example, salts of this compound include those that are protonated with an inorganic or organic acid to form a cation, along with the inorganic or organic acid's conjugate base as the anionic component of the salt.

[0204] A "solvate" refers to a complex formed by the bonding of a solvent molecule with a solute molecule or ion. The solvent is an organic compound, an inorganic compound, or a mixture of both. Some examples of solvents include, but are not limited to, methanol, N,N-dimethylformamide, tetrahydrofuran, dimethyl sulfoxide, and water. When the solvent is water, the solvate formed is a hydrate.

[0205] "Stereoisomers" refer to compounds that share the same atomic bonds but have different spatial arrangements of atoms. Stereoiomers include cis-trans isomers, E isomers, Z isomers, enantiomers, and diastereomers.

[0206] "Tautomers" refer to alternative molecular forms that differ only in the positions of electron bonds and / or protons of atoms (e.g., enol-keto tautomers and imine-enamine tautomers), or tautomers of heteroaryl groups (e.g., those containing the -N=C(H)-NH- ring configuration (e.g., pyrazole, imidazole, benzimidazole, triazole, tetrazole)). Those skilled in the art will recognize that other tautomer ring configurations are also possible.

[0207] It will be understood that the term “or its salt or solvate or stereoisomer” is intended to include any permutation of salts, solvates, and stereoisomers (e.g., solvates of pharmaceutically acceptable salts of stereoisomers of the compound of the subject).

[0208] In some embodiments, an “activatable” antibody refers to an antibody that exhibits a first level of binding to the target when inhibited, masked, and / or uncleaved, and a second level of binding to the target when uninhibited, unmasked, and / or cleaved, wherein the second level of target binding is higher than the first level of target binding. In some embodiments, access to the target by the activatable antibody occurs more frequently after cleavage within the cleavable portion (e.g., by one or more proteases).

[0209] "Pharmacologically effective dose" and "therapeutic effective dose" refer to the amount of a compound sufficient to treat one or more of a particular disorder or disease, or its symptoms, and / or to prevent the onset of the disease or disorder. With respect to neoplastic proliferative disorders, a pharmacochemically or therapeutically effective dose includes, among other things, an amount sufficient to shrink a tumor or reduce the rate of tumor growth.

[0210] "Preventive dose" is the amount of a pharmaceutical composition that, when administered to a subject, has the intended preventive effect, for example, the effect of preventing or delaying the onset (or recurrence) of a disease, disorder, or condition, or the effect of reducing the likelihood of the onset (or recurrence) of a disease, disorder, condition, or associated symptoms(s).

[0211] Complete therapeutic or preventive effects may not necessarily occur with a single dose, and may only occur after a series of doses. Therefore, an effective therapeutic or preventive dose may be administered in more than one dose.

[0212] The term "patient" refers to both human and non-human subjects (especially mammals).

[0213] As used herein, the terms “to treat” or “treatment” mean treating a disease or condition in a patient (e.g., a mammal (in particular, a human)), including: (a) preventing the onset of the disease or condition in question, e.g., preventive treatment; (b) improving the disease or condition in a patient, e.g., improving the disease or condition or causing its regression; (c) suppressing the disease or condition in a patient, e.g., slowing or halting the progression of the disease or condition; or (d) alleviating the symptoms of the disease or condition in a patient.

[0214] In some embodiments, the terms “to treat” or “treatment” exclude preventive treatment.

[0215] A “reaction partner” refers to a molecule or molecular part that specifically reacts with another reaction partner to produce a reaction product. An exemplary reaction partner includes cysteine ​​or serine of a sulfatase motif and formylglycine-producing enzyme (FGE), which react to form a reaction product of a converted aldehyde tag containing formylglycine (fGly) instead of cysteine ​​or serine in the motif. Another exemplary reaction partner includes the aldehyde (e.g., a reactive aldehyde group) of the fGly residue of the converted aldehyde tag and an “aldehyde-reactive reaction partner,” which includes an aldehyde-reactive group and the moiety of interest, which react to form a reaction product of a polypeptide having the moiety of interest conjugated to the polypeptide via the fGly residue.

[0216] The "N-terminus" refers to the terminal amino acid residue of a polypeptide that has a free amine group, while the amine groups of non-N-terminal amino acid residues usually form part of the polypeptide's covalent backbone.

[0217] The "C-terminus" refers to the terminal amino acid residue of a polypeptide that has a free carboxyl group, while the carboxyl groups of non-C-terminal amino acid residues usually form part of the covalent backbone of the polypeptide.

[0218] In relation to polypeptides or the amino acid sequences of polypeptides, the term "internal region" refers to the region of the polypeptide that is neither N-terminus nor C-terminus.

[0219] The term "subject" refers to both human and non-human subjects (especially mammalian subjects).

[0220] As used herein, the term “natural amino acid sequence” refers to the amino acid sequence of a polypeptide before it is modified to include modified amino acid residues.

[0221] The terms "amino acid analog" and "non-natural amino acid" are used interchangeably and include amino acid-like compounds that are structurally and / or in shape similar to one or more amino acids commonly found in natural proteins (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). Amino acid analogs also include natural amino acids with modified side chains or backbone. Amino acid analogs also include amino acid analogs that have the same stereochemistry as naturally occurring D- and L-type amino acid analogs. In some embodiments, amino acid analogs share the backbone structure and / or side chain structure of one or more natural amino acids, but the difference(s) is one or more modification groups within the molecule. Such modifications may include, but are not limited to, the substitution of an atom (e.g., N) for a related atom (e.g., S), the addition of a group (e.g., methyl, hydroxyl, etc.) or an atom (e.g., Cl, Br, etc.), the deletion of a group, the substitution of a covalent bond (e.g., a single bond to a double bond), or combinations thereof. For example, amino acid analogs include α-hydroxy acids, α-amino acids, etc.

[0222] The term "amino acid side chain" is used to refer to substituents attached to the α-carbon of an amino acid residue (including, for example, native amino acids, non-native amino acids, and amino acid analogs). Amino acid side chains may also include amino acid side chains described herein in the context of modified amino acids and / or conjugates.

[0223] The term "carbohydrate" is used to refer to monomeric units and / or polymers of monosaccharides, disaccharides, oligosaccharides, and polysaccharides. The term "sugar" is used to refer to small carbohydrates (e.g., monosaccharides, disaccharides). The term "carbohydrate derivative" includes compounds in which one or more functional groups of the target carbohydrate are substituted (replaced with any convenient substituent), modified (converted to another group using any convenient chemical reaction), or absent (e.g., removed or replaced with H). A variety of carbohydrates and carbohydrate derivatives are available and can be adapted for use in the subject compound and conjugate.

[0224] The terms "glycoside" or "glycosyl" refer to a sugar molecule or sugar group that is attached to a part via a glycosidic bond. For example, the part to which a glycoside is attached may be a cleavable linker as described herein. Glycosidic bonds can attach a glycoside to another part via various types of bonds (e.g., but not limited to O-glycosidic bonds (O-glycosides), N-glycosidic bonds (glycosylamines), S-glycosidic bonds (thioglycosides), or C-glycosidic bonds (C-glycosides or C-glycosyls). In some cases, a glycoside can be cleaved from the part to which it is attached, for example, by chemically mediated hydrolysis or enzymatic hydrolysis.

[0225] As used herein, “ROR antigen” refers to a member of the tyrosine-protein kinase transmembrane receptor (ROR) family, including members ROR1 and ROR2. In certain embodiments, the activatable ROR antibody has an antigen-binding site that specifically binds to ROR1 only. In other embodiments, the activatable ROR antibody has an antigen-binding site that specifically binds to ROR2 only. In other embodiments, the activatable ROR antibody has an antigen-binding site that is cross-reactive and specifically binds to both ROR1 and ROR2.

[0226] Accordingly, in one embodiment, the terms “ROR” or “ROR antigen” as used herein refer to ROR1 (e.g., human ROR1). In another embodiment, the terms “ROR” or “ROR antigen” as used herein refer to ROR2 (e.g., human ROR2). In yet another embodiment, the terms “ROR” or “ROR antigen” as used herein refer to both ROR1 and ROR2 (e.g., both human ROR1 and human ROR2).

[0227] The terms “antibody,” “immunoglobulin,” or “Ig” are used interchangeably herein and in their broadest sense, specifically encompassing, for example, polyclonal antibodies, monoclonal antibodies (including agonists, antagonists, neutralizing antibodies, and full-length monoclonal antibodies), antibody compositions having polyepitope or monoepitope specificity, recombinant antibodies, single-domain antibodies, single-specificity antibodies, multispecificity antibodies (including bispecificity antibodies), synthetic antibodies, chimeric antibodies, humanized antibodies, or human versions of antibodies having full-length heavy and / or light chains. Antibodies also include single antibody domains that retain ROR binding properties, as well as antibody fragments (and / or polypeptides containing antibody fragments). Non-limiting examples of antibody fragments include the antigen-binding region and / or effector region of an antibody, e.g., Fab, Fab', F(ab')2, Fv, scFv, (scFv)2, single-chain antibody molecules, bivariate domain antibodies, single variable domain antibodies, linear antibodies, V regions, multispecific antibodies formed from antibody fragments, F(ab)2, Fd, Fc, diabodies, didiabodies, disulfide-bonded Fv(dsFv), single-domain antibodies (e.g., nanobodies), or other fragments (e.g., fragments consisting of non-covalently bonded heavy and light chain variable regions). Generally speaking, the variable (V) region can be any preferred configuration of the immunoglobulin heavy chain (VH) and / or light chain (VL) variable regions. For example, antibodies also include tetrameric antibodies containing two heavy chain molecules and two light chain molecules, antibody light chain monomers, and antibody heavy chain monomers. Therefore, for example, the V region may be a dimer and may contain VH-VH, VH-VL, or VL-VL dimers that bind to the ROR. In any embodiment, the VH and VL regions may be covalently linked directly or via a linker to form a single-chain Fv (scFv). For ease of reference, scFv proteins are referred to herein as belonging to the category of “antibody fragments.” Another form of antibody fragment is a peptide containing one or more complementarity-determining regions (CDRs) of an antibody. CDRs (also called “minimum recognition units” or “hypervariable regions”) can be obtained by constructing polynucleotides encoding the one or more CDRs in question.Such polynucleotides can be prepared, for example, by using a polymerase chain reaction to synthesize the variable region using mRNA from antibody-producing cells as a template (see, for example: Larrick et al., Methods: A Companion to Methods in Enzymology, 2:106 (1991); Courtenay-Luck, “Genetic Manipulation of Monoclonal Antibodies,” in Monoclonal Antibodies Production, Engineering and Clinical Application, Ritter et al. (eds.), page 166, Cambridge University Press (1995); and Ward et al., “Genetic Manipulation and Expression of Antibodies,” in Monoclonal Antibodies: Principles and Applications, Birch et al., (eds.), page 137, Wiley-Liss, Inc. (1995)). Antibody fragments can be incorporated into, for example, single-domain antibodies, maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, the variable region (v-NAR) of novel antigen receptors, and bis-single-chain Fv regions (see, for example, Hollinger and Hudson, Nature Biotechnology, 23(9):1126-1136, 2005). In some embodiments, antibodies comprising VH and / or VL contain a light chain constant region and / or a heavy chain constant region (e.g., one or more constant regions, e.g., one or more IgG1, IgG2, IgG3, and / or IgG4 constant regions). In some embodiments, the antibody may contain any of the epitope-binding fragments described above. Antibodies described herein may be of any class of immunoglobulin molecules (e.g., IgG, IgE, IgM, IgD, and IgA) or any subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2).

[0228] The terms "humanized antibody" or "humanized immunoglobulin" refer to non-human (e.g., mouse or rabbit) antibodies that contain one or more amino acids (e.g., in the framework region, constant region, or CDR) that are substituted with amino acids at the corresponding positions in human antibodies. Generally, humanized antibodies produce a reduced immune response in the human host compared to the non-humanized version of the same antibody. Antibodies can be humanized using a variety of techniques known in the art, including, for example, CDR grafting (EP239,400; PCT Publication WO91 / 09967; U.S. Patents 5,225,539, 5,530,101, and 5,585,089), veneering or resurfacing (EP592,106, EP519,596, Padlan, Molecular Immunology 28(4 / 5):489-498(1991); Studnicka et al., Protein Engineering 7(6):805-814(1994); Roguska et al., PNAS 91:969-973(1994)), and chain shuffling (U.S. Patent 5,565,332). In certain embodiments, framework substitutions are identified by modeling the interaction between CDRs and framework residues to identify framework residues important for antigen binding, as well as by sequence comparison to identify abnormal framework residues at specific locations (see, for example, U.S. Patent No. 5,585,089, Riechmann et al., Nature 332:323 (1988)). Further methods for humanizing the antibodies intended for use in the present invention are described below: U.S. Patent Nos. 5,750,078; 5,502,167; 5,705,154; 5,770,403; 5,698,417; 5,693,493; 5,558,864; 4,935,496; and 4,816,567, as well as PCT Publications WO98 / 45331 and WO98 / 45332. In certain embodiments, the subject rabbit antibody may be humanized according to the methods described in US20040086979 and US20050033031. Thus, the above antibody may be humanized using methods well known in the art.

[0229] The term "chimeric antibody" refers to an antibody in which the light chain and heavy chain genes are constructed, typically through genetic engineering, from variable and constant region genes of antibodies belonging to different species. For example, the variable segment of a mouse monoclonal antibody gene may bind to human constant segments (e.g., gamma 1 and gamma 3). An example of a therapeutic chimeric antibody is a hybrid protein consisting of a variable domain or antigen-binding domain from a mouse antibody and a constant domain or effector domain from a human antibody, although domains from other mammalian species may be used.

[0230] As used herein, the term "single-specific" refers to an antibody having one or more binding sites, each of which binds to the same epitope of the same antigen.

[0231] When used in reference to an antibody, the term "multispecificity" means that the antibody can specifically bind to at least two distinct epitopes, for example, two binding sites formed by pairs of antibody heavy chain variable domains (VH) and antibody light chain variable domains (VL), or by VHH domains, which bind to different antigens or different epitopes on the same antigen. Such a bispecific antibody may have a 1+1 format (containing one binding site for a first antigen or epitope and one binding site for a second antigen or epitope). Other bispecific antibody formats include a 2+1 or 1+2 format (containing two binding sites for a first antigen or epitope and one binding site for a second antigen or epitope), or a 2+2 format (containing two binding sites for a first antigen or epitope and two binding sites for a second antigen or epitope). When a bispecific antibody contains two antigen-binding sites, each may bind to a different epitope. Such a bispecific antibody can bind to two different epitopes on the same antigen (e.g., epitopes on ROR).

[0232] The term “identical” or “identity” percentage, in the context of two or more nucleic acids or polypeptides, refers to two or more sequences or subsequences that are identical when compared and aligned to the greatest extent possible (with gaps introduced where necessary), and when none of the conserved amino acid substitutions are considered as part of sequence identity, or two or more sequences or subsequences that have a certain percentage of identical nucleotides or amino acid residues. Percentage identity can be measured using sequence comparison software or algorithms, or by visual inspection. Various algorithms and software that can be used to obtain alignment of amino acid or nucleotide sequences are well known in the art. These include, but are not limited to, the Basic Local Alignment Search Tool (BLAST), ALIGN, MegAlign, BestFit, GCG Wisconsin Package, and their variations. In some embodiments, two nucleic acids or polypeptides are substantially identical when compared and aligned to obtain the maximum correspondence, either when measured using a sequence comparison algorithm or by visual inspection. This means they have at least 70%, at least 75%, at least 80%, at least 85%, or at least 90%, and in some embodiments, at least 95%, 96%, 97%, 98%, or 99% nucleotide or amino acid residue identity. In some embodiments, the identity exists over a region of the amino acid sequence that is at least about 10 residues long, at least about 20 residues long, at least about 40–60 residues long, at least about 60–80 residues long, or any integer value in between. In some embodiments, the identity exists over a region longer than 60–80 residues (e.g., at least about 80–100 residues), and in some embodiments, the sequences are substantially identical over the entire length of the sequences being compared (e.g., the coding region of a target protein or antibody). In some embodiments, identity exists over a region of nucleotide sequence that is at least about 10 nucleotides long, at least about 20 nucleotides long, at least about 40–60 nucleotides long, at least about 60–80 nucleotides long, or any integer value in between.In some embodiments, the identity exists over a region longer than 60–80 bases (e.g., at least about 80–1000 bases or more), and in some embodiments, the sequence is substantially identical over the entire length of the sequence being compared (e.g., the nucleotide sequence encoding the protein of interest).

[0233] A "conservative amino acid substitution" is a substitution in which one amino acid residue is replaced by another amino acid residue having a side chain with similar chemical properties. Families of amino acid residues with similar side chains are generally defined in the art and include basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), non-charged side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), β-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). For example, the substitution of tyrosine with phenylalanine is a conservative substitution. Generally, conservative substitutions in the sequences of polypeptides, soluble proteins, and / or antibodies of this disclosure do not preclude binding to target binding sites of polypeptides, soluble proteins, or antibodies containing amino acid sequences. Methods for identifying non-precluding amino acid conservative substitutions are well known in the art.

[0234] The term “polypeptide” refers to a polymer of amino acids of any length. The polymer may be linear or branched, and may contain modified amino acids or non-amino acids (e.g., they may be substituted with non-amino acids). The term also includes amino acid polymers that are modified in nature or by intervention, e.g., by the formation of disulfide bonds, glycosylation, lipidization, acetylation, phosphorylation, or any other operation or modification, e.g., by binding to or conjugation (directly or indirectly) of a moiety, e.g., a labeling component or drug (e.g., a toxin). For example, polypeptides containing one or more analogs of amino acids (e.g., including non-natural amino acids), as well as other modifiers known in the art, are also included in the definition. Since the polypeptides of this disclosure may be based on antibodies or other members of the immunoglobulin superfamily, it will be understood that in some embodiments, the polypeptides may exist as single chains or single-chain dimers.

[0235] As used herein, “antigen” is a portion or molecule containing an epitope to which an antibody can bind. Thus, an antigen is to which an antibody can bind. In some embodiments, the antigen to which the antibody described herein binds is ROR (e.g., human ROR) or a fragment thereof.

[0236] As used herein, “epitope” is a term used in the art and refers to a local region of an antigen to which an antibody can bind. An epitope may be a linear epitope, or a conformational, nonlinear, or discontinuous epitope. For example, in the case of a polypeptide antigen, the epitope may be a sequence of amino acids in the polypeptide (a “linear” epitope), or it may consist of amino acids derived from two or more discontinuous regions of the polypeptide (a “conformational,” “nonlinear,” or “discontinuous” epitope) (e.g., human ROR). Generally, those skilled in the art will understand that linear epitopes may or may not depend on secondary, tertiary, or quaternary structure. For example, in some embodiments, an antibody binds to a group of amino acids regardless of whether the group is folded into a native three-dimensional protein structure. In other embodiments, the antibody requires that the amino acid residues constituting the epitope exhibit a specific conformation (e.g., bend, twist, turn, or fold) in order to recognize and bind to the epitope.

[0237] An antibody binds to an "epitope," a "similar epitope," or "the same epitope" as a reference antibody. The most widely used rapid method for determining whether two antibodies bind to identical, overlapping, or adjacent epitopes in three-dimensional space is a competition assay, which can be constructed in various forms (e.g., using either a labeled antigen or a labeled antibody). In some assays, the antigen is immobilized on a 96-well plate or expressed on the cell surface, and the ability of an unlabeled antibody to block the binding of a labeled antibody is measured using radioactive, fluorescent, or enzyme labeling.

[0238] As used herein, the terms “specifically bind,” “specifically recognize,” “immunely bind,” “selectively bind,” “immunely recognize,” and “immunely specific” are analogous terms in the context of antibodies and refer to molecules that bind to an antigen (e.g., an epitope), as understood by those skilled in the art.

[0239] In some embodiments, “specifically binding” means, for example, that a polypeptide or molecule interacts with an epitope, protein, or target molecule more frequently, more rapidly, longer, with greater affinity, or a combination of the above, than alternative substances (including related and unrelated proteins). For example, a molecule that specifically binds to an antigen may generally bind to other peptides or polypeptides with low affinity, as determined by: e.g., immunoassays, BIACORE®, KinExA 3000 instrument (Sapidyne Instruments, Boise, Idaho), OctetQK384 system (ForteBio, Menlo Park, California), or other assays known in the art. In some embodiments, an antibody or antigen-binding region binds to or specifically to an antigen with higher affinity than any cross-reactive antigen, as determined using experimental techniques (e.g., radioimmunoassays (RIA) and enzyme-linked immunosorbent assays (ELISA)). Typically, specific or selective reactions are at least twice the background signal or noise and may be more than 10 times the background. For a discussion of binding specificity, see, for example, Fundamental Immunology 332-36 (Paul ed., 2d ed. 1989). In some embodiments, the degree of binding of the antibody or antigen-binding region to "non-target" proteins is less than about 10% of the binding of the antibody or antigen-binding region to its target antigen, as determined, for example, by fluorescence-activated cell sorting (FACS) analysis or RIA. In some embodiments, a molecule that specifically binds to an antigen is K when the molecule binds to another antigen. A At least 2 logs, 2.5 logs, 3 logs, 4 logs, or more than K A It binds to the antigen. In some embodiments, the molecule that specifically binds to the antigen does not cross-react with other proteins. In another specific embodiment, the molecule that specifically binds to the antigen does not cross-react with other non-ROR proteins. In some embodiments, "specifically binds" means, for example, that the polypeptide or molecule binds to a protein or target at a concentration of about 0.1 mM or less (more commonly, less than about 1 μM) of K. Dmeans binding by. In some embodiments, "specifically binds" means that a polypeptide or molecule binds to a target with a K of at least about 0.1 μM or less, at least about 0.01 μM or less, or at least about 1 nM or less. D means binding by. Due to sequence identity between homologous proteins of different species, specific binding can include polypeptides or molecules that recognize proteins or targets of multiple species. Similarly, due to homology within a specific region of the polypeptide sequences of different proteins, specific binding can include polypeptides or molecules that recognize more than one protein or target. It can be seen that in some embodiments, a polypeptide or molecule that specifically binds to a first target may or may not specifically bind to a second target. Thus, "specific binding" does not necessarily (but may) require exclusive binding, e.g., binding to a single target. Thus, in some embodiments, a polypeptide or molecule can specifically bind to multiple targets. In some embodiments, multiple targets can be bound by the same antigen-binding site on the polypeptide or molecule. For example, in some embodiments, an antibody can include two identical antigen-binding sites, each of which specifically binds to the same epitope on two or more proteins. In another embodiment, the antibody is bispecific and can include at least two antigen-binding sites with different specificities. Generally, but not necessarily, references to "binding" mean "specific binding".

[0240] The term "binding affinity" generally refers to the total strength of non-covalent interaction between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen such as ROR). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of binding molecule X for its binding partner Y is generally the dissociation constant (K D) can be expressed as. Affinity can be measured by common methods known in the art (including methods described herein). Low affinity antibodies generally tend to bind slowly to antigens and dissociate easily, while high affinity antibodies generally tend to bind quickly to antigens and remain bound for longer. Various methods for measuring binding affinity are known in the art and any of them can be used for the purposes of this disclosure. In one embodiment, "K D " or "K D The value can be measured, for example, using the OctetQK384 system (ForteBio, Menlo Park, California) with biolayer interferometry (BLI). Alternatively, K D This may be measured, for example, by a radiolabeled antigen-binding assay (RIA) (Chen et al., (1999) J. Mol Biol 293:865-881) performed using the Fab version of the antibody of interest and its antigen, or by a surface plasmon resonance (SPR) assay using, for example, BIACORE®-2000 or BIACORE®-3000 (BIAcore, Inc., Piscataway, New Jersey). "On-rate" or "rate of association" or "rate of association" or "k" on ", as well as "off rate" or "dissociation rate" or "dissociation rate" or "k off These can also be determined using the same SPR or BLI methodology described herein, for example, using the OctetQK384 system (ForteBio, Menlo Park, California) or BIACORE®-2000 or BIACORE®-3000 (BIACORE®, Inc., Piscataway, New Jersey).

[0241] When used in the context of activatable ROR antibodies, the term "competition" refers to an antibody whose binding to an epitope or binding site is at least partially inhibited by the binding of another antibody in the presence of that antibody. Competition can be determined by assays that block or inhibit the specific binding of a reference molecule (e.g., a reference ligand, or a reference antigen-binding protein (e.g., a reference antibody)) to a common antigen (e.g., ROR). Various types of competitive binding assays can be used to determine whether a test antibody competes with a reference molecule for binding to ROR (e.g., human ROR). Examples of assays that can be used include solid-phase direct or indirect radioimmunoassays (RIAs), solid-phase direct or indirect enzyme immunoassays (EIAs), sandwich competition assays (e.g., see Stahli et al., (1983) Methods in Enzymology 9:242-253); solid-phase direct biotin-avidin EIA (e.g., see Kirkland et al., (1986) J.Immunol. 137:3614-3619 or Cheung et al., (1990) Virology 176:546-552); solid-phase direct labeling assays, solid-phase direct labeling sandwich assays (e.g., see Harlow and Lane, (1988) Antibodies, A Laboratory Manual, Cold Spring Harbor Press); solid-phase direct labeling RIA using I-125 labeling (e.g., see Morel et al.) al., (1988) Molec. Immunol. 25:7-15); and directly labeled RIA (Moldenhauer et al., (1990) Scand. (J. Immunol. 32:77-82). Typically, such assays involve the use of purified antigen (e.g., ROR (e.g., human ROR)) bound to a solid surface or cell having either an unlabeled test antigen-binding protein (e.g., test ROR antibody or ADC) or a labeled reference antigen-binding protein (e.g., reference TF antibody or ADC). Competitive inhibition can be measured by determining the amount of labeling bound to the solid surface or cell in the presence of the test antigen-binding protein.Typically, test antigen-binding proteins are present in excess. Antibodies identified in a competitive assay (competitive antibodies) include antibodies that bind to the same epitope as the reference antibody, and / or antibodies that bind to adjacent epitopes (e.g., analogous or duplicate epitopes) that are sufficiently close to the epitope bound to the reference antibody that would cause steric hindrance to the antibody. Typically, when competitive antibodies are present in excess, the specific binding of the reference antibody to the common antigen will be inhibited by at least 20%, e.g., at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75%. In some embodiments, binding is inhibited by at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% or more.

[0242] In this specification, the terms “constant region” and “constant domain” are used interchangeably and are well-known antibody technical terms, referring to a portion of an antibody, for example, the carboxyl-terminal portion of the light and / or heavy chain that is not directly involved in the binding of the antibody to an antigen but can exhibit various effector functions (e.g., interaction with Fc receptors). The term includes portions of immunoglobulin molecules that have amino acid sequences that are generally more conserved than the immunoglobulin variable region.

[0243] The "effector function" of an antibody refers to the biological activity resulting from the antibody's Fc region (e.g., the native sequence Fc region or the amino acid sequence variant Fc region), which varies depending on the antibody isotype. Examples of antibody effector functions include C1q binding and complement-dependent cytotoxicity; Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g., B cell receptors); and B cell activation.

[0244] In this specification, the term “Fc region” is used to define the C-terminal region of an immunoglobulin heavy chain (including, for example, the native sequence Fc region, the recombinant Fc region, and the variant Fc region). While the boundaries of the Fc region of an immunoglobulin heavy chain can vary, the human IgG heavy chain Fc region is often defined as extending from the amino acid residue at position Cys226 (EU numbering system) or Pro230 (EU numbering system) to the carboxyl terminus. The C-terminal lysine of the Fc region (residue 447 in the EU numbering system) can be removed, for example, during antibody production or purification, or by recombinant operations on the nucleic acid encoding the antibody heavy chain. Examples of Fc region sequences are provided below (CH2 domain = bold text, CH3 domain = underlined text): [ka]

[0245] A "functional Fc region" possesses "effector functions" of a native sequence Fc region. Exemplary "effector functions" include C1q binding; complement-dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; and downregulation of cell surface receptors (e.g., B cell receptors, BCRs). Such effector functions typically require the Fc region to be combined with a binding region or binding domain (e.g., an antibody variable region or domain) and can be evaluated using the various assays disclosed.

[0246] "Natural sequence Fc regions" contain amino acid sequences identical to those of Fc regions found in nature and have not been manipulated, modified, and / or altered by humans (e.g., isolated, purified, selected, included, or combined with other sequences (e.g., variable region sequences)). Natural sequence human Fc regions include natural sequence human IgG1 Fc regions (non-A and A allotypes); natural sequence human IgG2 Fc regions; natural sequence human IgG3 Fc regions; and natural sequence human IgG4 Fc regions; as well as their natural variants.

[0247] A "variant Fc region" includes an amino acid sequence that differs from the amino acid sequence of the natural sequence Fc region by at least one amino acid modification (e.g., substitution, addition, or deletion), preferably one or more amino acid substitutions. In some embodiments, the variant Fc region has at least one amino acid substitution compared to the natural sequence Fc region or the Fc region of the parent polypeptide, for example, having about 1 to about 10 amino acid substitutions, preferably about 1 to about 5 amino acid substitutions, in the natural sequence Fc region or the Fc region of the parent polypeptide. The variant Fc regions described herein may have at least about 80% homology to the natural sequence Fc region and / or the Fc region of the parent polypeptide, or at least about 90% homology (e.g., at least about 95% homology). The variant Fc regions described herein may have loss of effector function (e.g., silent Fc (also referred to herein as "sFc")).

[0248] In some embodiments, the variant Fc region includes an alanine (Ala, A) residue at position Leu234 (L234) according to the EU numbering system, an alanine (Ala, A) residue at position Leu235 (L235) according to the EU numbering system, and a lysine (Lys, K) residue at position Pro329 (P329) according to the EU numbering system (also referred to herein as "LALAPK" or "L234A / L235A / P329K").

[0249] An example variant Fc region ("silent Fc") sequence is shown below (CH2 domain = (bold text with underlined amino acid changes), CH3 domain = underlined text). [ka]

[0250] Additionally or alternatively, the variant Fc region has reduced potential immunogenicity. In further embodiments, the variant Fc region comprises a glutamic acid (Glu,E) residue at position Asp356(D356) according to the EU numbering system, a glutamic acid (Glu,E) residue at position Glu357(E357) according to the EU numbering system, and a methionine (Met,M) residue at position Leu358(L358) according to the EU numbering system (also referred to herein as "EEM" or "D356E / E357E / L358M").

[0251] When used in relation to antibodies, the term "heavy chain" refers to a polypeptide chain of approximately 50–70 kDa, with an amino-terminal variable region containing approximately 120–130 or more amino acids and a carboxy-terminal constant region containing one or more constant regions. A "heavy chain" can refer to different types (e.g., alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ)) based on the amino acid sequence of the constant region, which give rise to antibodies of the IgA, IgD, IgE, IgG, and IgM classes (e.g., subclasses of IgG, e.g., IgG1, IgG2, IgG3, and IgG4), respectively.

[0252] As used herein, the term “light chain,” when used in relation to antibodies, refers to a polypeptide chain of approximately 25 kDa, where the amino-terminal portion contains a variable region of approximately 100 to approximately 110 or more amino acids, and the carboxy-terminal portion contains a constant region. The approximate length of a light chain is 211 to 217 amino acids. Based on the amino acid sequence of the constant region, there are two different types (e.g., kappa (κ) or lambda (λ)). Light chain amino acid sequences are well known in the art. In one embodiment, the “chain” (e.g., heavy chain or light chain) is the molecule (e.g., polypeptide) itself. In another embodiment, the “chain” (e.g., heavy chain or light chain) is a part of the molecule (e.g., polypeptide) and is conjugated directly or indirectly to the rest of the molecule (e.g., polypeptide), for example.

[0253] The terms “antigen-binding fragment,” “antigen-binding domain,” and “antigen-binding region” refer to a portion of an antibody (e.g., a CDR) that interacts with an antigen and contains amino acid residues that confer specificity and affinity to the antigen to the binding fragment or region. As used herein, “antigen-binding fragment” includes an “antibody fragment” that contains a portion of an antibody (e.g., one or more CDRs, e.g., the antigen-binding region or variable region of an antibody).

[0254] The antibodies described herein include, but are not limited to, synthetic antibodies, monoclonal antibodies, recombinant antibodies, multispecific antibodies (e.g., including bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, intrabodies, single-chain Fv(scFv) (e.g., including monospecific, bispecific, etc.), camelized antibodies, Fab fragments, F(ab') fragments, disulfide-linked Fv(sdFv), anti-idiotype (anti-Id) antibodies, and any of the epitope-linked fragments described above.

[0255] In some embodiments, the antibodies described herein include an immunoglobulin molecule and an immunologically active portion of the immunoglobulin molecule (for example, a molecule containing one or more antigen-binding sites that bind to ROR).

[0256] The antibodies described herein may be any type of immunoglobulin molecule (e.g., IgG, IgE, IgM, IgD, IgA, or IgY), any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2), or any subclass (e.g., IgG2a, or IgG2b). In some embodiments, the activatable ROR antibody described herein is an IgG antibody (e.g., human IgG), or a class thereof (e.g., human IgG1, IgG2, IgG3, or IgG4), or a subclass thereof.

[0257] In some embodiments, the antibody is a four-chain antibody unit containing two heavy (H) chain / light (L) chain pairs. In further embodiments, the amino acid sequences of the H chains are identical, and the amino acid sequences of the L chains are identical. In other embodiments, the amino acid sequences of the H chains are different from each other. Additionally or alternatively, the amino acid sequences of the L chains are different from each other. For example, the antibody contains a first H / L chain pair and a second H / L chain pair, where the first H / L chain pair binds to a ROR antigen and the second H / L chain pair binds to another ROR antigen or a non-ROR antigen. In some embodiments, the antibody is a two-chain antibody unit containing a VHH-VHH pair. In further embodiments, the amino acid sequences of the VHHs are identical. In other embodiments, the amino acid sequences of the VHHs are different from each other. For example, the antibody contains a first VHH and a second VHH, where the first VHH binds to a ROR antigen and the second VHH binds to another ROR antigen or a non-ROR antigen. In some embodiments, the H chain and / or L chain include a constant region, e.g., a human constant region. In some embodiments, the L chain constant region of such an antibody is a kappa light chain constant region or a lambda light chain constant region, e.g., a human kappa light chain constant region or a human lambda light chain constant region. In some embodiments, the H chain constant region of such an antibody includes a gamma heavy chain constant region, e.g., a human gamma heavy chain constant region. In some embodiments, such an antibody includes an IgG constant region, e.g., a human IgG constant region (e.g., IgG1, IgG2, IgG3, and / or IgG4 constant regions).

[0258] As used herein, “ROR antibody” and “antibody that binds to ROR” are used interchangeably and refer to an antibody that preferentially binds to ROR. An antibody or fragment thereof may preferentially bind to ROR (e.g., human ROR), meaning that the antibody or fragment binds to ROR (e.g., human ROR) with a higher affinity than it would to an unrelated control protein. For example, an antibody or fragment may specifically recognize or bind to ROR or a portion thereof. “Specific binding” means that the ROR antibody or fragment binds to ROR with an affinity at least 5, 10, 15, 20, 25, 50, 100, 250, 500, 1000, or 10,000 times higher than its affinity to an unrelated control protein (e.g., chicken egg white lysozyme). In some embodiments, a ROR antibody or fragment thereof may bind to ROR substantially exclusively (e.g., ROR can be distinguished from other known polypeptides by a measurable difference in binding affinity). In some embodiments, the ROR antibody may react with ROR sequences other than human ROR sequences (e.g., cynomolgus monkey ROR sequences).

[0259] The terms “variable region” and “variable domain” are used interchangeably to refer to parts of the light and heavy chains of an antibody, which are generally located at the amino termini of the light and heavy chains, with the heavy chain being approximately 120–130 amino acids long and the light chain approximately 100–110 amino acids long, and are used for each antibody’s binding and specificity to its antigen. The variable region of the heavy chain is referred to herein as “VH.” The variable region of the light chain is referred to herein as “VL.” The term “variable” refers to the fact that the sequences of specific segments of the variable region differ significantly between antibodies. The V region mediates antigen binding and defines the specificity of a particular antibody to its antigen. However, variability is not evenly distributed across the 110-amino acid range of the variable region. Instead, the V region consists of a less variable (e.g., relatively invariant) stretch of approximately 15–30 amino acids called the framework region (FR), which is separated by a shorter region of greater variability (e.g., extreme variability) called the “hypervariable region” or “complementarity-determining region.” The variable regions of the heavy and light chains each contain four frameworks (FR1, FR2, FR3, and FR4), primarily employing a β-sheet configuration linked by three hypervariable regions. These regions form loops, linking the β-sheet structures and, in some cases, forming parts of the β-sheet structure. The hypervariable regions of each chain are held together in close proximity by the frameworks and, together with the hypervariable regions of other chains, participate in the formation of the antibody's antigen-binding site (see, for example, Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, (1991)). The constant region does not directly participate in the binding of the antibody to the antigen but exerts various effector functions (e.g., the antibody's involvement in antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cell-mediated cytotoxicity (CDC)). The sequences of the variable regions differ significantly between different antibodies. Sequence variability is concentrated in the CDR, but the less variable parts within the variable region are called the framework region (FR). The CDRs of the light and heavy chains are primarily involved in the interaction of the antigen with the antibody.In certain embodiments, the variable region is the human variable region.

[0260] As used herein, the terms “hypervariable region,” “HVR,” “HV,” “complementarity-determining region,” and “CDR” refer to regions of antibody variable regions whose sequences are hypervariable and / or form structurally defined loops. Generally, antibodies contain six hypervariable regions: three in VH (H1 or VH CDR1, H2 or VH CDR2, and H3 or VH CDR3) and three in VL (L1 or VL CDR1, L2 or VL CDR2, and L3 or VL CDR3). Several descriptions of hypervariable regions have been used and are incorporated herein. Kabat CDRs are based on sequence diversity and are the most widely used (see, for example, Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)). Chothia, on the other hand, refers to the location of structural loops (e.g., Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). The ends of the Chothia CDR-H1 loop vary from H32 to H34 depending on the length of the loop, if numbered according to Kabat numbering rules (this is because if the Kabat numbering scheme places insertions at H35A and H35B and neither 35A nor 35B exists, the loop ends at 32; if only 35A exists, the loop ends at 33; and if both 35A and 35B exist, the loop ends at 34). The AbM hypervariable region is Kabat This represents the compromise between the CDR and the Chothia structural loop and is used in Oxford Molecular's AbM antibody modeling software (see, e.g., Martin, in Antibody Engineering, Vol. 2, Chapter 3, Springer Verlag). The hypervariable regions of "contact" are based on an analysis of available complex crystal structures. The residues derived from each of these hypervariable regions or CDRs are shown below.

[0261] A universal numbering system has been developed and widely adopted: the ImMunoGeneTics (IMGT®) information system (Lefranc et al., Dev.Comp.Immunol.27(1):55-77(2003)). IMGT® is an integrated information system specifically for human and other vertebrate immunoglobulins (IG), T cell receptors (TR), and major histocompatibility complexes (MHC). In this specification, CDRs are referred to in terms of both their amino acid sequence and their position within the light or heavy chain. The “position” of CDRs within the structure of immunoglobulin variable regions is conserved across species and resides within structures called loops; therefore, by using a numbering system that aligns the variable region sequences of structural features, CDRs and framework residues can be easily identified. This information can be used when transplanting and substituting CDR residues from certain immunoglobulins into acceptor frameworks, typically derived from human antibodies. The additional numbering system (AHon) has been developed by: Honegger and Pluckthun, J.Mol.Biol.309:657-670 (2001). For example, correspondences between numbering systems, including the Kabat numbering and the IMGT® proprietary numbering system, are well known to those skilled in the art (see, for example, Kabat, supra; Chothia and Lesk, cited above; Martin, supra; Lefranc et al., cited above), and also shown below. The various systems known in the art or described herein represent various ways of describing CDRs, and when used to define the same antibody, they are often considered equivalent. The exemplary system shown herein combines Kabat and Chothia. [Table 12]

[0262] The hypervariable regions are as follows: In VL, 24-36 or 24-34 (L1), 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3); in VH, 26-35 or 26-35A (H1), 50-65 or 49-65 (H2), and the "extended hypervariable regions" of 93-102, 94-102, or 95-102 (H3).

[0263] As used herein, the term “isolated” means that the target compound is present in an environment different from the environment in which the compound naturally occurs. “Isolated” means that the target compound is in a substantially concentrated sample and / or that the target compound is partially or substantially purified.

[0264] The terms “ROR-mediated disease,” “ROR-mediated disorder,” and “ROR-mediated condition” are used interchangeably to refer to any disease, disorder, or condition associated with or characterized by ROR-expressing cells (e.g., TF-expressing tumor cells). ROR-mediated diseases include, but are not limited to, cancers that express or overexpress ROR.

[0265] In any embodiment of this specification, the term “tumor” means the growth or proliferation of any neoplastic cells, whether malignant or benign, and all precancerous and cancerous cells and tissues.

[0266] The terms "cancer" and "cancerous" typically refer to or describe a physiological condition in mammals characterized by uncontrolled cell growth.

[0267] The terms “ADC” or “activatable-ADC” (the terms are used interchangeably) refer to an antibody-drug conjugate, which in the context of this invention refers to an activatable ROR antibody conjugated to another portion containing a drug, as described herein.

[0268] As used herein, “drug” refers to a compound having biological activity, such as a cytotoxic compound (e.g., a cytotoxic small molecule, a cytotoxic synthetic peptide, etc.).

[0269] Examples of drugs include small molecule drugs (e.g., cancer chemotherapy drugs). For example, if the polypeptide is an antibody (or a fragment thereof) having specificity for tumor cells, the antibody can be modified to include modified amino acids as described herein, and then this can be conjugated into a cancer chemotherapy drug. Cancer chemotherapy drugs include non-peptidic (e.g., non-proteinic) compounds that reduce the proliferation of cancer cells, and these include cytotoxic agents and cell proliferation inhibitors. Non-limiting examples of chemotherapy drugs include alkylating agents, nitrosoureas, antimetabolites, antitumor antibiotics, plant (vinca) alkaloids, and steroid hormones. Peptide compounds can also be used.

[0270] Suitable cancer chemotherapy agents include drastatin and its active analogs and derivatives, as well as auristatin and its active analogs and derivatives (e.g., monomethyl auristatin D (MMAD), monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), etc.). See, for example, WO96 / 33212, WO96 / 14856, and US6,323,315. For example, drastatin 10 or auristatin PE may be included in the activatable ROR-ADC of this disclosure. Suitable cancer chemotherapy agents include maytansinoids and their active analogs and derivatives (e.g., see below: EP1391213, and Liu et al (1996) Proc. Natl. Acad. Sci. USA 93:8618-8623); duocalmycin and its active analogs and derivatives (e.g., including the synthetic analogs KW-2189 and CB1-TM1); and benzodiazepines and their active analogs and derivatives (e.g., pyrrolobenzodiazepine (PBD)).

[0271] Drugs that act to reduce cell proliferation are known and widely used in the art. Such drugs include alkylating agents (e.g., nitrogen mustard, nitrosourea, ethyleneimine derivatives, alkyl sulfonates, and triazenes), which include, but are not limited to, mechloretamine, cyclophosphamide (CYNOTAN®), melphalan (L-sarcolicin), carmustine (BCNU), lomustine (CCNU), semustine (methyl-CCNU), streptozosin, chlorozotosin, uracil mustard, chlormethine, ifosfamide, chlorambucil, pipobromane, triethylenemelamine, triethylenethiophosphoramine, busulfan, dacarbazine, and temozolomide.

[0272] Antimetabolites include folate analogs, pyrimidine analogs, purine analogs, and adenosine deaminase inhibitors, which include, but are not limited to, cytarabine (CYTOSAR-U®), cytosine arabinoside, fluorouracil (5-FU), floxuridine (FudR), 6-thioguanine, 6-mercaptopurine (6-MP), pentostatin, 5-fluorouracil (5-FU), methotrexate, 10-propargyl-5,8-dideazafolate (PDDF, CB3717), 5,8-dideazatetrahydrofolate (DDATHF), leucovorin, fludarabine phosphate, pentostatin, and gemcitabine.

[0273] Suitable natural products and their derivatives (e.g., vinca alkaloids, antitumor antibiotics, enzymes, lymphokines, and epipodophyllotoxins) include, but are not limited to, Ara-C, paclitaxel (TAXOL®), docetaxel (TAXOTERE®), deoxycoformycin, mitomycin C, L-asparaginase, azathioprine; brequinal; alkaloids (e.g., vincristine, vinblastine, vinorelbine, vindesine, etc.); podophyllotoxins (e.g., etoposide, teniposide, etc.); antibiotics (e.g., ant Lacycline, daunorubicin hydrochloride (daunomycin, rubidomycin, serubicin), idarubicin, doxorubicin, epirubicin, and morpholino derivatives, etc.; phenoxyzombine cyclopeptides (e.g., dactinomycin); basic glycopeptides (e.g., bleomycin); anthraquinone glycosides (e.g., plicamycin (mitramycin)); anthracendions (e.g., mitoxantrone, etc.); azilinopyrroloindolediones (e.g., mitomycin); macrocyclic immunosuppressants (e.g., cyclosporine, FK-506 (tacrolimus, Prograf), rapamycin, etc.).

[0274] Other antiproliferative cytotoxic drugs include navelben, CPT-11, anastrazole, letrazole, capecitabine, reloxafine, cyclophosphamide, ifosamide, and droloxafine.

[0275] Microtubule agonists with antiproliferative activity are also suitable for use, including but not limited to: allocolchicine (NSC 406042), halichondrin B (NSC 609395), colchicine (NSC 757), colchicine derivatives (e.g., NSC 33410), dorustatin 10 (NSC 376128), meitansine (NSC 153858), rhizoxin (NSC 332598), paclitaxel (TAXOL®), TAXOL® derivatives, docetaxel (TAXOTERE®), thiocolchicine (NSC 361792), tritylcisterine, vinblastine sulfate, vincristine sulfate, natural and synthetic eoptilons (including, but not limited to, eoptilon A, eoptilon B, discodermorid; estramustine, nocodazole, etc.).

[0276] Suitable hormone regulators and steroids (including synthetic analogs) for use include, but are not limited to, the following: corticosteroids (e.g., prednisone, dexamethasone), estrogens and pregestins (e.g., hydroxyprogesterone caproate, medroxyprogesterone acetate, megestrol acetate, estradiol, clomiphene, tamoxifen), and corticosteroids (e.g., aminoglutethimide; 17α-ethinylestradiol; diethylstilbestrol; testosterone, fluoxymesterone, dromostanolone propionate). Examples include testolactone, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianicene, hydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesterone acetate, leuprolide, flutamide (DROGENIL®), toremifene (FARESTON®), goserelin (ZOLADEX®), etc. Estrogen stimulates proliferation and differentiation, and therefore compounds that bind to estrogen receptors are used to inhibit this activity. Corticosteroids can inhibit T cell proliferation.

[0277] Other suitable chemotherapeutic agents include: metal complexes (e.g., cisplatin (cis-DDP), carboplatin, etc.); urea (e.g., hydroxyurea); hydrazine (e.g., N-methylhydrazine); epidophyllotoxin; topoisomerase inhibitors; procarbazine; mitoxantrone; leucovorin; tegafur, etc. Other targeted antiproliferative agents include: immunosuppressants (e.g., mycophenolic acid, thalidomide, desoxysperguarin, azasporin, leflunomide, mizoribine, azaspiran (SKF105685), gefitinib (IRESSA®, (ZD 1839, 4-(3-chloro-4-fluorophenylamino)-7-methoxy-6-(3-(4-morpholinyl)propoxy)quinazoline), etc.).

[0278] Taxanes are suitable for use. "Taxane" includes paclitaxel and active taxane derivatives or prodrugs. "Paclitaxel" (which should be understood herein to include analogs, formulations, and derivatives (e.g., docetaxel, TAXOL®, TAXOTERE® (formulations of docetaxel), 10-desacetyl analogs of paclitaxel, and 3'N-desbenzoyl-3'Nt-butoxycarbonyl analogs of paclitaxel)) can be readily prepared using methods known to those skilled in the art (see below: WO94 / 0 7882, WO94 / 07881, WO94 / 07880, WO94 / 07876, WO93 / 23555, WO93 / 10076, U.S. Patent Nos. 5,294,637, 5,283,253, 5,279,949, 5,274,137, 5,202,448, 5,200,534, 5,229,529, and EP590,267), or available from various commercial suppliers (e.g., Sigma). Chemical Co., St. Louis, Missouri (T7402 from Taxus brevifolia, or T-1912 from Taxus yannanensis). It should be understood that paclitaxel refers not only to the commonly available chemical forms of paclitaxel, but also to analogs and derivatives (e.g., TAXOTERE® docetaxel as described herein) and paclitaxel conjugates (e.g., paclitaxel-PEG, paclitaxel-dextran, or paclitaxel-xylose).

[0279] The term "taxane" includes various known derivatives (e.g., both hydrophilic and hydrophobic derivatives). Taxane derivatives include, but are not limited to, the galactose and mannose derivatives described in International Patent Application WO99 / 18113; piperazino and other derivatives described in WO99 / 14209; taxane derivatives described in WO99 / 09021, WO98 / 22451, and U.S. Patent No. 5,869,680; 6-thio derivatives described in WO98 / 28288; sulfenamide derivatives described in U.S. Patent No. 5,821,263; and taxol derivatives described in U.S. Patent No. 5,415,869. Furthermore, it includes prodrugs of paclitaxel, which include, but are not limited to, those described in WO98 / 58927, WO98 / 13059, and U.S. Patent No. 5,824,701.

[0280] Suitable biological response modifiers include, but are not limited to, (1) tyrosine kinase (RTK) activity inhibitors; (2) serine / threnion kinase activity inhibitors; (3) tumor-associated antigen antagonists such as antibodies that specifically bind to tumor antigens; (4) apoptosis receptor agonists; (5) interleukin-2; (6) IFN-α; (7) IFN-γ; (8) colony-stimulating factors; and (9) angiogenesis inhibitors.

[0281] An "effective dose" is generally a quantity sufficient to reduce the severity and / or frequency of symptoms, eliminate symptoms and / or underlying causes, prevent the onset of symptoms and / or underlying causes, and / or improve or correct damage resulting from or associated with a disease, disorder, or condition. In some embodiments, the effective dose is a therapeutically effective dose.

[0282] As used herein, the term “therapeutic dose” refers to the amount of the antibody or ADC described herein that is sufficient to reduce and / or improve the severity and / or duration of a particular disease, disorder, or condition and / or symptoms associated therewith. The therapeutic dose of a drug, including a therapeutic agent, may be the amount necessary to (i) reduce or improve the advancement or progression of a given disease, disorder, or condition; (ii) reduce or improve the recurrence, progression, or onset of a given disease, disorder, or condition; and / or (iii) improve or enhance the therapeutic effect of another treatment (e.g., a treatment other than the administration of the antibody or ADC described herein). The “therapeutic dose” of a substance / molecule / drug of this disclosure (e.g., an activatable ROR antibody or an activatable ADC) may vary based on the number of factors that induce the desired response in an individual (e.g., the individual’s medical condition, age, sex, and weight, as well as the capacity of the substance / molecule / drug). The therapeutic dose includes the amount in which any toxic or adverse effects of the substance / molecule / drug outweigh the therapeutically beneficial effects. In some embodiments, the term “therapeutic dose” refers to the amount of antibody or other agent (e.g., drug) that is effective in “treating” a disease, disorder, or condition in a subject or mammal.

[0283] In some embodiments, the drug is a microtubule agonist having antiproliferative activity (e.g., a mytansinoid). In some embodiments, the drug is an antimitotic agent (e.g., auristatin or an active auristatin analog or a derivative thereof). In some embodiments, the drug is a DNA alkylating agent.

[0284] As used herein, the term “pharmaceutically acceptable” means that it is approved by a federal or state regulatory authority or is listed in the United States Pharmacopeia, the European Pharmacopeia, or any other generally accepted pharmacopoeia used for animals (in particular, humans).

[0285] "Excipients" include carriers, excipients, preservatives, or stabilizers that are non-toxic to cells or mammals to which they are exposed at the dosage and concentration used, and may be included, for example, to affect stability, increase the volume of the formulation, or impart a therapeutic effect to the active ingredient in the final dosage form (e.g., to promote absorption, reduce viscosity, or increase solubility). "Excipients" may be organic or inorganic natural or synthetic components that are combined with the active ingredient to facilitate the use of the active ingredient (e.g., combined with the active ingredient to facilitate the administration of the active ingredient to a target). Examples of excipients include: buffers (e.g., phosphoric acid, citrate, and other organic acids); antioxidants (e.g., ascorbic acid); low molecular weight (e.g., fewer than 10 amino acid residues) polypeptides; proteins (e.g., serum albumin, gelatin, or immunoglobulin); hydrophilic polymers (e.g., polyvinylpyrrolidone); amino acids (e.g., glycine, glutamine, asparagine, arginine, or lysine); monosaccharides, disaccharides, and other carbohydrates (e.g., glucose, mannose, or dextrin); chelating agents (e.g., EDTA); sugar alcohols (e.g., mannitol or sorbitol); salt-forming counterions (e.g., sodium); and / or nonionic surfactants (e.g., TWEEN®, polyethylene glycol (PEG), and PLURONICS®). The term “excipient” may also refer to diluents, adjuvants (e.g., Freund’s adjuvants (complete or incomplete)), excipients, or vehicles used in the administration of therapeutic agents. Such excipients may be sterile liquids (e.g., water and oils of petroleum, animal, plant, or synthetic origin, e.g., peanut oil, soybean oil, mineral oil, sesame oil, etc.). Water is an exemplary excipient when the composition (e.g., a pharmaceutical composition) is administered intravenously. Saline solution, aqueous dextrose, and glycerol solutions may also be used as liquid excipients (especially for injection solutions).Suitable excipients (e.g., pharmaceutical excipients) include: starch, glucose, lactose, sucrose, gelatin, malt, rice, wheat flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, skimmed dried milk, glycerol, propylene, glycol, water, ethanol, etc. In any embodiment, the composition may also contain small amounts of wetting agents, emulsifiers, or pH buffers. The composition may take the form of a solution, suspension, emulsion, tablet, pill, capsule, powder, sustained-release formulation, etc. Oral compositions (e.g., formulations) may contain standard excipients (e.g., pharmaceutical-grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc.). Examples of suitable excipients are described in Remington: The Science and Practice of Pharmacy (2020) (Elsevier Science, Amsterdam, Netherlands). For example, a pharmaceutical compound may contain an effective or therapeutically effective amount of activatable ROR-ADC, for example in an isolated or purified form, along with a suitable amount of excipients, to provide a form for appropriate administration to the target. The formulation must be suitable for the mode of administration.

[0286] The terms "approximately" and "about" refer to variations of 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, and 1% or less of a given value or range.

[0287] As used herein, comparative terms, such as reduction, decrease, increase, or any of these grammatical variations, can refer to a particular variation from a reference value. In some embodiments, such variation can refer to a value that is about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% higher than the reference value, or about 1x, 2x, 3x, 4x, 5x, 10x, 20x, 30x, 40x, or 100x higher. In some embodiments, such variation can refer to about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the reference value.

[0288] As used in this disclosure and claims, the singular forms with “a,” “an,” and “the” include the plural form unless the context clearly indicates otherwise.

[0289] In some embodiments, the terms “first,” “second,” “third,” “fourth,” and synonyms in the names of components are used to distinguish and identify one or more components that share a particular identity in those names. For example, “first antibody” and “second antibody” are used to distinguish two antibodies.

[0290] Wherever embodiments are described herein using the phrase "including," it is understood that other similar embodiments are also provided, described using the phrases "consisting of" and / or "essentially consisting of." Wherever embodiments are described herein using the phrase "essentially consisting of," it is understood that other similar embodiments are also provided, described using the phrase "consisting of."

[0291] The term "between" or "between" refers to a range that includes both A and B, as used in phrases such as "between A and B" or "between A and B."

[0292] When the term "and / or" is used in this specification in phrases such as "A and / or B," it is intended to include both A and B, A or B, A (alone), and B (alone). Similarly, when the term "and / or" is used in phrases such as "A, B and / or C," it is intended to include each of the embodiments of A, B and C, A, B or C, A or C, A or B, B or C, A and C, A and B, B and C, A (alone), B (alone), and C (alone).

[0293] The terms "optional" or "at your discretion" mean that the situations described below may or may not occur, so these descriptions include both cases where the situations occur and cases where they do not.

[0294] Before further describing the present invention, it should be understood that the present invention is not limited to the specific embodiments described and can, for example, be modified. It should also be understood that the terms used herein are for the sole purpose of describing specific embodiments and are not intended to limit the scope of the present invention, as it is limited only by the appended claims.

[0295] Where a range of values ​​is provided, each intervening value (up to one-tenth of the lower limit unless otherwise explicitly stated in the context), the range between the upper and lower limits of that range, and any other specified or intervening values ​​within the specified range are understood to be included in the present invention. The upper and lower limits of these smaller ranges may independently be included in the smaller range and are further included in the present invention, subject to any restrictions specifically excluded within the described range. Where a described range includes one or both of the limit values, a range excluding one or both of the included limit values ​​is also included in the present invention.

[0296] For clarity, it is understood that certain features of the Invention described in the context of separate embodiments may also be provided in combination in a single embodiment. Conversely, for brevity, various features of the Invention described in the context of a single embodiment may also be provided separately or as components of any suitable combination. All combinations of embodiments relating to the Invention are specifically encompassed in the Invention and are disclosed herein as if every possible combination were individually and explicitly disclosed, insofar as such combinations encompass subject matter that is, for example, a stable compound (e.g., a compound that can be produced, isolated, characterized, and tested for biological activity). In addition, all subcombinations of various embodiments and their elements (e.g., elements of chemical groups enumerated in embodiments describing such variables) are also specifically encompassed in the Invention and are disclosed herein as if every possible subcombination were individually and explicitly disclosed herein.

[0297] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which the present invention pertains. Methods and materials similar to or equivalent to those described herein may also be used in carrying out or testing the present invention, but only methods and materials are described here. All publications referenced herein are incorporated herein by reference to disclose and describe the methods and / or materials cited in those publications.

[0298] For clarity, it is understood that certain features of the invention described in the context of separate embodiments may also be provided in combination in a single embodiment. Conversely, for brevity, various features of the invention described in the context of a single embodiment may also be provided individually or in any suitable combination of components.

[0299] Only publications disclosed prior to the filing date of this application are presented herein. Nothing in this specification should be construed as admitting that the present invention does not have prior rights to such publications on the grounds of prior invention. Furthermore, the publication dates provided may differ from the actual publication dates and may need to be verified.

[0300] 7.2 ROR-ADC Antibodies that bind to tyrosine protein kinase membrane receptors (RORs) (also referred herein as “ROR antibodies,” “anti-ROR antibodies,” “ROR Ab,” “Ab,” or “antibodies”), such as activatable ROR antibodies (also referred herein as “activatable ROR antibodies,” “activatable anti-ROR antibodies,” “activatable ROR Ab,” “activatable Ab,” or “activatable antibodies”), and drugs may be directly or indirectly linked to each other via pyridazine-pyrrolo coupling moieties to form ROR-ADCs as described herein, such as activatable ROR-ADCs. In certain embodiments, ROR antibodies, such as activatable ROR antibodies, and two or more drugs or activators are linked to each other via one or more functional groups and covalent bonds. For example, one or more functional groups and covalent bonds may include branched linkers as described herein.

[0301] A target portion (e.g., a drug or active agent) can be conjugated to a ROR antibody, such as an activatable ROR antibody, at any desired site on the antibody. Accordingly, this disclosure provides an activatable ROR antibody having a site T conjugated to two or more sites on the antibody (e.g., a site at or near the C-terminus of the antibody, a site at or near the N-terminus of the antibody, and a site between the C-terminus and N-terminus of the antibody (e.g., an internal site of the antibody)). Combinations of the above conjugation portions are also possible.

[0302] In certain embodiments, the conjugate of the present disclosure comprises two (or more) drugs or activators conjugated to the α-carbon of an amino acid residue of a ROR antibody, for example, an activatable ROR antibody. In other words, the conjugate comprises a ROR antibody, for example, an activatable ROR antibody, in which the side chain of an amino acid residue in the antibody is modified and conjugated to two (or more) drugs or activators (e.g., conjugated to two drugs or activators via a branched linker as described herein). For example, the conjugate comprises a ROR antibody, for example, an activatable ROR antibody, in which the α-carbon of an amino acid residue in the antibody is modified and conjugated to two drugs or activators (e.g., conjugated to two drugs or activators via a branched linker as described herein).

[0303] Embodiments of the present disclosure include conjugates in which a ROR antibody, for example, an activatable ROR antibody, is conjugated to two or more parts (e.g., three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, or twenty or more parts). These parts can be conjugated to a ROR antibody, for example, an activatable ROR antibody, at multiple sites within the antibody. In some embodiments, two parts can be conjugated to a single amino acid residue of the activatable ROR antibody. For example, two parts can be conjugated to the same amino acid residue of the activatable ROR antibody. In other embodiments, two parts are conjugated to the first amino acid residue of the activatable ROR antibody, and the other two parts are conjugated to the second amino acid residue of the activatable ROR antibody. For example, an activatable ROR antibody can be conjugated to the first and second portions by the first amino acid residue, and to the third and fourth portions, etc., by the second amino acid residue. In some cases, two or more amino acid residues of an activatable ROR antibody are each conjugated to a pair of portions (e.g., two portions), and each pair of portions is conjugated to an activatable ROR antibody via a branched linker as described herein. In some cases, one amino acid residue in an activatable ROR antibody is conjugated to a pair of portions via a branched linker as described herein. In other examples, two or more amino acid residues (e.g., 3, 4, 5, 6, 7, 8, 9, or 10 or more amino acid residues) in an activatable ROR antibody are each conjugated to a pair of portions via a branched linker as described herein.

[0304] The ROR antibody conjugated to the target portion, for example, one or more amino acid residues of an activatable ROR antibody, may be native amino acids, non-natural amino acids, or a combination thereof. For example, the conjugate may include a target portion (e.g., a drug or activator) conjugated to a native amino acid residue of an activatable ROR antibody. In other examples, the conjugate may include a target portion conjugated to a non-natural amino acid residue of an activatable ROR antibody. The target portion can be conjugated to an activatable ROR antibody with a single native or non-natural amino acid residue as described above. One or more native or non-natural amino acid residues in an activatable ROR antibody can be conjugated to the target portion as described herein. For example, two (or more) amino acid residues (e.g., native or non-natural amino acid residues) in an activatable ROR antibody may each be conjugated to two portions via a branched linker, resulting in multiple sites in the activatable ROR antibody being conjugated to the target portion.

[0305] As described herein, a ROR antibody, for example, an activatable ROR antibody, can be conjugated to two or more target moieties. In certain embodiments, the target moiety is a payload, for example, a chemical entity (e.g., a drug, an activator, or a detectable label). For example, a drug (or activator, e.g., a cytokine) can be conjugated to an activatable ROR antibody, or in other embodiments, a detectable label can be conjugated to an activatable ROR antibody. In other embodiments, different payload combinations can be conjugated to an activatable ROR antibody. Thus, embodiments of the present disclosure include, but are not limited to, conjugations of an activatable ROR antibody with two or more drugs, conjugations of an activatable ROR antibody with two or more activators (e.g., cytokines), conjugations of an activatable ROR antibody with two or more detectable labels, and combinations thereof.

[0306] In certain embodiments, a ROR antibody, e.g., an activatable ROR antibody, and a moiety of interest (e.g., a drug or active agent) are conjugated via a conjugation moiety. For example, the ROR antibody, e.g., an activatable ROR antibody, and the moiety of interest may each be bound (e.g., covalently) to the conjugation moiety, thereby indirectly linking the antibody and the moiety of interest via the conjugation moiety. In some cases, the conjugation moiety includes a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl compound, or a derivative of a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl compound. For example, a general scheme for binding the coupling moiety of interest to a ROR antibody, e.g., an activatable ROR antibody, via a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety is shown in the general reaction scheme below. The hydrazinyl-indolyl and hydrazinyl-pyrrolo-pyridinyl conjugation moieties are also referred to herein as the hydrazino-iso-pictet-spengler (HIPS) conjugation moiety and the aza-hydrazino-iso-pictet-spengler (azaHIPS) conjugation moiety, respectively. [ka]

[0307] An ROR antibody, for example, is conjugated to an activatable ROR antibody (e.g., conjugated to an activatable ROR antibody via a linker described herein) and contains a moiety of interest (e.g., a drug or bioactive agent), where n is an integer from 1 to 4. As shown in the reaction scheme above, the conjugation moiety (e.g., a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety) is bound to two or more drugs or bioactive agents R. An ROR antibody containing a 2-formylglycine residue (fGly), for example, an activatable ROR antibody, reacts with the conjugation moiety to produce an activatable ROR antibody conjugate, whereby two or more drugs or bioactive agents are bound to the antibody via the conjugation moiety.

[0308] As described herein, the moiety can be any of a variety of moieties (e.g., but not limited to, chemical entities such as detectable labels, or drugs or bioactive agents). R’ and R” can each independently be any desired substituent (e.g., but not limited to, hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl). Z can be CR 21 , NR 22 , N, O, or S, and R 21 and R 22 are each independently selected from any of the substituents described for R’ and R” above.

[0309] Other hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moieties are also possible, as shown in the conjugates and compounds described herein. For example, a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety may be bonded (e.g., covalently) to two or more linkers. Thus, embodiments of the present disclosure include a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety bonded to two or more drugs or activators, each via a corresponding linker. Thus, the conjugates of the present disclosure may include two or more linkers, each linker bonding the corresponding drug or activator to the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety. Therefore, the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety and two or more linkers may be considered as a "branched linker" as a whole, where the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety is bound to two or more "branches," and each branch contains a linker to which the drug or active agent is bound.

[0310] The same or different payload combinations can be conjugated to a ROR antibody, such as an activatable ROR antibody, via a branched linker. In certain embodiments, two payloads (e.g., drugs, activators, or detectable labels) bound to the branched linker are the same payload (e.g., drugs, activators, or detectable labels). For example, the first branch of the branched linker may be bound to a payload (e.g., a drug, activator, or detectable label), while the second branch of the branched linker may be bound to the same payload (e.g., a drug, activator, or detectable label) as the first branch.

[0311] In other embodiments, the two payloads (e.g., drugs, activators, or detectable labels) coupled to the branched linker are different payloads (e.g., drugs, activators, or detectable labels). For example, the first branch of the branched linker may be coupled to a first payload (e.g., a first drug, activator, or detectable label), while the second branch of the branched linker may be coupled to a second payload (e.g., a second drug, activator, or detectable label) that is different from the first payload (e.g., a first drug, activator, or detectable label) coupled to the first branch.

[0312] In some embodiments, when two different drugs or activators are coupled to a branched linker, the drugs or activators may be selected from drugs and activators that have a synergistic therapeutic effect. "Synergistic," "synergistic effect," or "synergistic effect" means a therapeutic effect greater than the sum of the effects of the drugs or activators taken separately. For example, in some cases, using two different drugs or activators coupled to a branched linker may result in a lower therapeutically effective concentration at which both payloads act, thereby increasing the overall potency of the ADC.

[0313] In some embodiments, when two different drugs or activators are bound to a branched linker, the drugs or activators may be selected from drugs and activators that provide an enhanced therapeutic effect compared to when the drugs or activators are used separately. For example, the drugs or activators may contribute to a higher efficacy of drug delivery by the ADC (e.g., some payloads (e.g., iRGD peptides) may increase extravasation into tissue and enhance tumor invasion).

[0314] In some embodiments, when two different drugs or activators are bound to a branched linker, the drugs or activators may be selected from drugs and activators that use different mechanisms of action. In some cases, targeting multiple pathways may provide a reduction in tumor drug resistance. Examples of payload combinations include, but are not limited to, cytotoxic drugs, immunomodulatory molecules that activate or inhibit immune cell populations, cytokines, hormones, and radioisotope-loaded chelating agents.

[0315] In some embodiments, two different payloads are coupled to a branched linker, and the payloads can be selected from a combination of a drug or active agent and a detectable label. For example, the first payload may be a detectable label used as an imaging agent or tracer for detecting the location of the ADC in vivo, while the second payload may be a drug or active agent that produces a therapeutic effect.

[0316] Various embodiments of linkers capable of conjugating hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moieties to drugs or active ingredients are described in detail herein. For example, in some examples, the linker is a cleavable linker (e.g., a cleavable linker) as described herein.

[0317] In certain embodiments, a ROR antibody, such as an activatable ROR antibody, may be conjugated to two or more target sites, and one or more amino acids of the antibody are modified before conjugation to the target sites. Modifying one or more amino acids of the antibody may produce an antibody containing one or more reactive groups suitable for conjugation to the target sites. In some cases, a ROR antibody, such as an activatable ROR antibody, may contain one or more modified amino acid residues to provide one or more reactive groups suitable for conjugation to the target sites (e.g., two or more sites bound to a conjugation site (such as a hydrazinyl-indol or hydrazinyl-pyrrolo-pyridinyl conjugation site)). For example, the amino acids of the antibody may be modified to include a reactive aldehyde group (e.g., a reactive aldehyde). Reactive aldehydes may be contained in an "aldehyde tag" or "ald tag," which, as used herein, refers to an amino acid sequence derived from a sulfatase motif (e.g., L(C / S)TPSR, SEQ ID NO: 99), which has been converted by the action of formylglycinase (FGE) to contain a 2-formylglycine residue (referred herein to as "fGly"). The fGly residue produced by FGE is also called "formylglycine." In other words, the term “aldehyde tag” as used herein refers to a “converted” sulfatase motif (e.g., a sulfatase motif in which a cysteine ​​or serine residue has been converted to fGly by the action of FGE (e.g., L(fGly)TPSR, SEQ ID NO: 123). A converted sulfatase motif can be generated from an amino acid sequence containing an “unconverted” sulfatase motif (e.g., an unconverted sulfatase motif in which a cysteine ​​or serine residue has not been converted to fGly by FGE but can be converted, e.g., an unconverted sulfatase motif having sequence: LCTPSR, SEQ ID NO: 100).In relation to the action of formylglycine-producing enzymes (FGE) on sulfatase motifs, “conversion” refers to biochemical modifications of a sulfatase motif from a cysteine ​​or serine residue to a formylglycine (fGly) residue (e.g., Cys to fGly, or Ser to fGly). Further embodiments of aldehyde tags and their use in site-directed protein modification are described in U.S. Patents 7,985,783 and 8,729,232, the disclosures of which are incorporated herein by reference.

[0318] In some cases, to generate a conjugate, an ROR antibody containing an fGly residue, such as an activatable ROR antibody, may be conjugated to the desired moiety by reacting fGly with a compound (e.g., a compound containing a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety as described above). For example, an fGly-containing ROR antibody may be brought into contact with a reaction partner under conditions suitable for providing conjugation of two or more drugs to the ROR antibody. In some cases, the reaction partner may contain a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety as described above. For example, two or more drugs or activators may be bound to the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety. In some cases, the drug or activator is bound to the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety (for example, covalently bound to hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl), and each drug or activator is bound to the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety via a corresponding linker. Thus, the fGly residue conjugated to the desired moiety after the reaction is referred to herein as 'fGly'.

[0319] In certain embodiments, the conjugate of the present disclosure comprises a ROR antibody, e.g., an activatable ROR antibody, having at least one amino acid residue bound to two or more target moieties (e.g., drugs or activators). To construct a conjugate, the amino acid residue of a ROR antibody, e.g., an activatable ROR antibody, may be modified and then bound to two or more drugs or activators bound to the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety as described above. In certain embodiments, the amino acid residue of a ROR antibody, e.g., an activatable ROR antibody, is a cysteine ​​or serine residue modified to an fGly residue as described above. In certain embodiments, a modified amino acid residue (e.g., an fGly residue) is conjugated to two or more drugs or activators containing a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety as described above, providing a conjugate of the present disclosure in which two or more drugs or activators are conjugated to an antibody via the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety. As used herein, the term "fGly" refers to an amino acid residue of the antibody conjugated to the moiety of interest (e.g., a drug or activator).

[0320] In certain embodiments, the conjugate comprises a ROR antibody having at least one amino acid residue conjugated to a branched linker described herein, for example, an activatable ROR antibody, which is then conjugated to two or more drugs or activators. For example, the conjugate may comprise a ROR antibody having at least one amino acid residue (fGly') conjugated to the above-described target moiety (e.g., a drug or activator), for example, an activatable ROR antibody.

[0321] In some embodiments, the ROR-ADC (e.g., an activatable ROR-ADC) is a. Antibodies that bind to tyrosine protein kinase membrane receptors (RORs) (e.g., activatable antibodies that bind to RORs); and b. Two or more drugs conjugated via a linker to the pyridazine-pyrrolo coupling portion. Includes.

[0322] In some embodiments, the ROR-ADC is a conjugate of formula (I): [ka] (In the formula, Ab represents an antibody that binds to ROR, for example, an activatable antibody that binds to ROR; Z 1 , Z 2 , Z 3 and Z 4 CR 4 , N and CL B -W 2 Each is independently selected from and at least one Z 1 , Z 2 , Z 3 and Z 4 CL B -W 2 and; R 1 However, selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, substituted heterocyclyl, R 2 and R 3 Each of these is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, or R 2 and R 3However, they can be arbitrarily joined in a ring to form a 5-membered or 6-membered heterocycline. Each R 4 However, independently selected from hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, L A It is the first linker; L B It is the second linker; s is an integer between 1 and 10; W 1 It is the first drug; W 2 (This is the second drug.)

[0323] The substituents associated with the conjugate of formula (I) are described in more detail below.

[0324] In a particular embodiment, Z 1 , Z 2 , Z 3 , and Z 4 However, each is independent of CR 4 , N and CL B -W 2 Selected from, at least one Z 1 , Z 2 , Z 3 , and Z 4 However, CL B -W 2 In a particular embodiment, Z 1 However, CR 4 In a particular embodiment, Z 1 However, N is the case. In a particular embodiment, Z 1 However, CL B -W 2 In a particular embodiment, Z2 is CR 4 In certain embodiments, Z 2 is N. In certain embodiments, Z 2 is C-L<00�0957>-W 2 In certain embodiments, Z 3 is CR 4 In certain embodiments, Z 3 is N. In certain embodiments, Z 3 is C-L B -W<000096]4>In certain embodiments, Z 4 is CR 4 In certain embodiments, Z 4 is N. In certain embodiments, Z 4 is C-L B -W 2 In some embodiments, each of Z 1 Z 3 and Z 4 is CR 4 In some embodiments, Z 3 is C-L B -W 2 In some embodiments, various combinations of Z

[0325] are possible. For example, in some cases, Z 1 Z 2 Z 3 Z 4 are possible. For example, in some cases, Z 1 is C-L B -W 2 Z 2 is CR 4 In some cases, Z 3 is CR 4 In some cases, Z 4 is CR 4 In some cases, Z 1 is CR 4 In some cases, Z 2 is C-L B -W 2 In some cases, Z 3 is CR 4 In some cases, Z 4 is CR 4In some cases, Z 1 However, CR 4 And Z 2 However, CR 4 And Z 3 However, CL B -W 2 And Z 4 However, CR 4 In some cases, Z 1 However, CR 4 And Z 2 However, CR 4 And Z 3 However, CR 4 And Z 4 However, CL B -W 2 That is the case.

[0326] In a particular embodiment, R 1 R is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R 1 is hydrogen. In a particular embodiment, R 1 is alkyl or substituted alkyl (e.g., C 1-6 Alkyl or C 1-6 Substituting alkyl, or C 1-4 Alkyl or C 1-4 Substituting alkyl, or C 1-3 Alkyl or C 1-3 It is a substituted alkyl. In certain embodiments, R 1 is an alkenyl or substituted alkenyl (e.g., C 2-6 Alkenil or C 2-6 Substituting alkenyl, or C 2-4 Alkenil or C 2-4 Substituting alkenyl, or C 2-3 Alkenil or C 2-3 It is a substituted alkenyl. In certain embodiments, R 1 is an alkynyl or substituted alkynyl (e.g., C 2-6 Alkenil or C2-6 Substituting alkenyl, or C 2-4 Alkenil or C 2-4 Substituting alkenyl, or C 2-3 Alkenil or C 2-3 It is a substituted alkenyl. In certain embodiments, R 1 is an aryl or substituted aryl (for example, C 5-8 Aryl or C 5-8 Substituting aryls (for example, C5 aryl or C5-substituted aryl, or C6 aryl or C6-substituted aryl). In certain embodiments, R 1 This refers to heteroaryl or substituted heteroaryl (e.g., C 5-8 heteroaryl or C 5-8 A substituted heteroaryl, for example, a C5 heteroaryl or a C5-substituted heteroaryl, or a C6 heteroaryl or a C6-substituted heteroaryl. In certain embodiments, R 1 This refers to cycloalkyl or substituted cycloalkyl (e.g., C 3-8 Cycloalkyl or C 3-8 Substitutive cycloalkyls, for example, C 3-6 Cycloalkyl or C 3-6 Substituted cycloalkyl, or C 3-5 Cycloalkyl or C 3-5 It is a substituted cycloalkyl. In certain embodiments, R 1 This refers to heterocyclines or substituted heterocyclines (e.g., C 3-8 Heterocycline or C 3-8 Substituting heterocyclyls, e.g., C 3-6 Heterocycline or C 3-6 Substituted heterocyclyl, or C 3-5 Heterocycline or C 3-5 It is a substituted heterocyclyl.

[0327] In a particular embodiment, R 2 and R 3Each of these is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, or R 2 and R 3 However, they can be arbitrarily joined in a ring to form a 5-membered or 6-membered heterocycline.

[0328] In a particular embodiment, R 2 is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R 2 is hydrogen. In a particular embodiment, R 2 is alkyl or substituted alkyl (e.g., C 1-6 Alkyl or C 1-6 Substituting alkyl, or C 1-4 Alkyl or C 1-4 Substituting alkyl, or C 1-3 Alkyl or C 1-3 It is a substituted alkyl. In certain embodiments, R 2 is methyl. In certain embodiments, R 2 is an alkenyl or substituted alkenyl (e.g., C 2-6 Alkenil or C 2-6 Substituting alkenyl, or C 2-4 Alkenil or C 2-4 Substituting alkenyl, or C2-3 Alkenil or C 2-3 It is a substituted alkenyl. In certain embodiments, R 2 is an alkynyl or substituted alkynyl. In certain embodiments, R 2 is an alkoxy or substituted alkoxy. In certain embodiments, R 2 is an amino acid or a substituted amino acid. In certain embodiments, R 2 is a carboxyl or carboxyl ester. In certain embodiments, R 2 is an acyl or acyloxy. In certain embodiments, R 2 is acylamino or aminoacyl. In certain embodiments, R 2 is an alkylamide or a substituted alkylamide. In certain embodiments, R 2 is a sulfonyl. In certain embodiments, R 2 is a thioalkoxy or substituted thioalkoxy. In certain embodiments, R 2 is an aryl or substituted aryl (for example, C 5-8 Aryl or C 5-8 Substituting aryls (for example, C5 aryl or C5-substituted aryl, or C6 aryl or C6-substituted aryl). In certain embodiments, R 2 This refers to heteroaryl or substituted heteroaryl (e.g., C 5-8 heteroaryl or C 5-8 A substituted heteroaryl, for example, a C5 heteroaryl or a C5-substituted heteroaryl, or a C6 heteroaryl or a C6-substituted heteroaryl. In certain embodiments, R 2 This refers to cycloalkyl or substituted cycloalkyl (e.g., C 3-8 Cycloalkyl or C 3-8 Substitutive cycloalkyls, for example, C 3-6 Cycloalkyl or C 3-6 Substituted cycloalkyl, or C 3-5 Cycloalkyl or C 3-5 It is a substituted cycloalkyl. In certain embodiments, R 2 This refers to heterocyclines or substituted heterocyclines (e.g., C3-6 Heterocycline or C 3-6 Substituted heterocyclyl, or C 3-5 Heterocycline or C 3-5 It is a substituted heterocyclyl.

[0329] In a particular embodiment, R 3 is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R 3 is hydrogen. In a particular embodiment, R 3 is alkyl or substituted alkyl (e.g., C 1-6 Alkyl or C 1-6 Substituting alkyl, or C 1-4 Alkyl or C 1-4 Substituting alkyl, or C 1-3 Alkyl or C 1-3 It is a substituted alkyl. In certain embodiments, R 3 is methyl. In certain embodiments, R 3 is an alkenyl or substituted alkenyl (e.g., C 2-6 Alkenil or C 2-6 Substituting alkenyl, or C 2-4 Alkenil or C 2-4 Substituting alkenyl, or C 2-3 Alkenil or C 2-3 It is a substituted alkenyl. In certain embodiments, R 3 is an alkynyl or substituted alkynyl. In certain embodiments, R 3 is an alkoxy or substituted alkoxy. In certain embodiments, R 3 is an amino acid or a substituted amino acid. In certain embodiments, R 3is a carboxyl or carboxyl ester. In certain embodiments, R 3 is an acyl or acyloxy. In certain embodiments, R 3 is acylamino or aminoacyl. In certain embodiments, R 3 is an alkylamide or a substituted alkylamide. In certain embodiments, R 3 is a sulfonyl. In certain embodiments, R 3 is a thioalkoxy or substituted thioalkoxy. In certain embodiments, R 3 is an aryl or substituted aryl (for example, C 5-8 Aryl or C 5-8 Substituting aryls (for example, C5 aryl or C5-substituted aryl, or C6 aryl or C6-substituted aryl). In certain embodiments, R 3 This refers to heteroaryl or substituted heteroaryl (e.g., C 5-8 heteroaryl or C 5-8 A substituted heteroaryl, for example, a C5 heteroaryl or a C5-substituted heteroaryl, or a C6 heteroaryl or a C6-substituted heteroaryl. In certain embodiments, R 3 This refers to cycloalkyl or substituted cycloalkyl (e.g., C 3-8 Cycloalkyl or C 3-8 Substitutive cycloalkyls, for example, C 3-6 Cycloalkyl or C 3-6 Substituted cycloalkyl, or C 3-5 Cycloalkyl or C 3-5 It is a substituted cycloalkyl. In certain embodiments, R 3 This refers to heterocyclines or substituted heterocyclines (e.g., C 3-8 Heterocycline or C 3-8 Substituting heterocyclyls, e.g., C 3-6 Heterocycline or C 3-6 Substituted heterocyclyl, or C 3-5 Heterocycline or C 3-5 It is a substituted heterocyclyl.

[0330] In a particular embodiment, R 2 and R 3 Both are methyl.

[0331] In a particular embodiment, R 2 and R 3 They may optionally be bonded in a ring to form a 5-membered or 6-membered heterocycline. In certain embodiments, R 2 and R 3 These are bonded in a ring to form a 5-membered or 6-membered heterocycline. In certain embodiments, R 2 and R 3 These are bonded in a ring to form a 5-membered heterocycline. In certain embodiments, R 2 and R 3 These molecules bond in a ring to form a six-membered heterocycline.

[0332] In a specific method of operation, each R 4 These are independently selected from hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

[0333] Each R 4 The various possibilities are detailed below. In a particular embodiment, R 4 is hydrogen. In a particular embodiment, each R 4 is hydrogen. In a particular embodiment, R 4 is a halogen (e.g., F, Cl, Br, or I). In certain embodiments, R 4 is F. In a particular embodiment, R 4 is Cl. In certain embodiments, R 4 is Br. In certain embodiments, R 4 is I. In a particular embodiment, R4 is alkyl or substituted alkyl (e.g., C 1-6 Alkyl or C 1-6 Substituting alkyl, or C 1-4 Alkyl or C 1-4 Substituting alkyl, or C 1-3 Alkyl or C 1-3 It is a substituted alkyl. In certain embodiments, R 4 is methyl. In certain embodiments, R 4 is an alkenyl or substituted alkenyl (e.g., C 2-6 Alkenil or C 2-6 Substituting alkenyl, or C 2-4 Alkenil or C 2-4 Substituting alkenyl, or C 2-3 Alkenil or C 2-3 It is a substituted alkenyl. In certain embodiments, R 4 is an alkynyl or substituted alkynyl. In certain embodiments, R 4 is an alkoxy or substituted alkoxy. In certain embodiments, R 4 is an amino acid or a substituted amino acid. In certain embodiments, R 4 is a carboxyl or carboxyl ester. In certain embodiments, R 4 is an acyl or acyloxy. In certain embodiments, R 4 is acylamino or aminoacyl. In certain embodiments, R 4 is an alkylamide or a substituted alkylamide. In certain embodiments, R 4 is a sulfonyl. In certain embodiments, R 4 is a thioalkoxy or substituted thioalkoxy. In certain embodiments, R 4 is an aryl or substituted aryl (for example, C 5-8 Aryl or C 5-8 Substituting aryls, for example, C5 aryls or C5-substituted aryls, or C6 aryls or C6-substituted aryls (for example, phenyl or substituted phenyl). In certain embodiments, R 4 This refers to heteroaryl or substituted heteroaryl (e.g., C5-8 heteroaryl or C 5-8 A substituted heteroaryl, for example, a C5 heteroaryl or a C5-substituted heteroaryl, or a C6 heteroaryl or a C6-substituted heteroaryl. In certain embodiments, R 4 This refers to cycloalkyl or substituted cycloalkyl (e.g., C 3-8 Cycloalkyl or C 3-8 Substitutive cycloalkyls, for example, C 3-6 Cycloalkyl or C 3-6 Substituted cycloalkyl, or C 3-5 Cycloalkyl or C 3-5 It is a substituted cycloalkyl. In certain embodiments, R 4 This refers to heterocyclines or substituted heterocyclines (e.g., C 3-8 Heterocycline or C 3-8 Substituting heterocyclyls, e.g., C 3-6 Heterocycline or C 3-6 Substituted heterocyclyl, or C 3-5 Heterocycline or C 3-5 It is a substituted heterocyclyl.

[0334] In a particular embodiment, L A This is the first linker. Examples of linkers that can be used in the conjugates of this disclosure are described in detail below.

[0335] In a particular embodiment, L B This is a second linker. Examples of linkers that can be used in the conjugates of this disclosure are detailed below.

[0336] In a particular embodiment, W 1 is the first drug (or first active agent). Examples of drugs and active agents that may be used in the conjugates of this disclosure are described below.

[0337] In a particular embodiment, W 2is a second drug (or second active agent). Examples of drugs and active agents that may be used in the conjugates of this disclosure are described below.

[0338] In certain embodiments, Ab represents an antibody that binds to ROR ("ROR antibody"). In certain embodiments, Ab represents an activatable antibody that binds to ROR ("activatable ROR antibody"). In certain embodiments, Ab comprises one or more fGly' residues as described herein. In certain embodiments, the ROR antibody is bound to the remainder of the conjugate via fGly' residues as described herein. In certain embodiments, the activatable ROR antibody is bound to the remainder of the conjugate via fGly' residues as described herein. Examples of ROR antibodies that can be used in the conjugates of this disclosure are detailed below.

[0339] In certain embodiments, the conjugate of formula (I) is a first linker, L A Includes the first linker, L A The first linker, L, can be used to conjugate the desired first portion (e.g., a first drug or active agent) to a ROR antibody, e.g., an activatable ROR antibody, via the conjugation portion. A The first linker, L A The hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety can be bound to the first drug. The hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety is bound to the first linker, L A Therefore, the first drug can be used to conjugate a ROR antibody, such as an activatable ROR antibody.

[0340] For example, as shown in equation (I) above, L AIt is bound to Ab via a conjugation moiety, and therefore Ab is linked to linker L via a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety. A It is indirectly bound to L. As described above, Ab is a ROR antibody, for example, an activatable ROR antibody, and thereby L A However, it is bound to ROR antibodies, such as activatable ROR antibodies, via a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety, for example, linker L A However, it indirectly binds to ROR antibodies, such as activatable ROR antibodies, via hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moieties.

[0341] The first linker L in the subject conjugate and compound A Any suitable linker can be used. In a particular embodiment, the first linker L A However, it may also contain a group selected from alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acylamino, alkylamide, substituted alkylamide, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, the first linker L A It may contain alkyl or substituted alkyl groups. In certain embodiments, the first linker L A It may contain an alkenyl or substituted alkenyl group. In certain embodiments, the first linker L A It may contain an alkynyl or substituted alkynyl group. In certain embodiments, the first linker L A It may contain an alkoxy or substituted alkoxy group. In certain embodiments, the first linker L A It may contain an amino or substituted amino group. In certain embodiments, the first linker L AIt may contain a carboxyl or carboxyl ester group. In certain embodiments, the first linker L A It may contain an acylamino group. In a particular embodiment, the first linker L A It may contain an alkylamide or a substituted alkylamide group. In certain embodiments, the first linker L A It may contain an aryl or substituted aryl group. In certain embodiments, the first linker L A It may contain a heteroaryl or substituted heteroaryl group. In certain embodiments, the first linker L A It may contain a cycloalkyl or a substituted cycloalkyl group. In certain embodiments, the first linker L A It may contain a heterocyclyl or a substituted heterocyclyl group.

[0342] In a particular embodiment, the first linker L A The polymer may include a polymer. For example, the polymer may include polyalkylene glycol and its derivatives (e.g., polyethylene glycol, methoxypolyethylene glycol, polyethylene glycol homopolymer, polypropylene glycol homopolymer, copolymer of ethylene glycol and propylene glycol (e.g., the homopolymer and copolymer are unsubstituted or have one end substituted with an alkyl group)), polyvinyl alcohol, polyvinyl ethyl, ether, polyvinylpyrrolidone, and combinations thereof. In certain embodiments, the polymer is polyalkylene glycol. In certain embodiments, the polymer is polyethylene glycol. Other linkers are also usable, as shown in the conjugates and compounds described in more detail below.

[0343] In some embodiments, L A This is the first linker, described by the following formula: -(L 1 ) a -(L 2 ) b -(L 3 )c -(L 4 ) d -(L 5 ) e -(L 6 ) f - (In the formula, L 1 , L 2 , L 3 , L 4 , L 5 and L 6 Each of these is an independent linker subunit, and a, b, c, d, e, and f are each independently 0 or 1, provided that at least one of a, b, c, d, e, and f is 1).

[0344] In certain embodiments, the sum of a, b, c, d, e, and f is between 1 and 6. In certain embodiments, the sum of a, b, c, d, e, and f is 1. In certain embodiments, the sum of a, b, c, d, e, and f is 2. In certain embodiments, the sum of a, b, c, d, e, and f is 3. In certain embodiments, the sum of a, b, c, d, e, and f is 4. In certain embodiments, the sum of a, b, c, d, e, and f is 5. In certain embodiments, the sum of a, b, c, d, e, and f is 6. In certain embodiments, a, b, c, d, e, and f are each 1. In certain embodiments, a, b, c, d, and e are 1, and f is 0. In certain embodiments, a, b, c, and d are each 1, and e and f are each 0. In certain embodiments, a, b, and c are each 1, and d, e, and f are each 0. In certain embodiments, a and b are each 1, and c, d, e, and f are each 0. In certain embodiments, a is 1, and b, c, d, e, and f are each 0.

[0345] In a particular embodiment, linker subunit L 1 However, it is bonded to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety (for example, as shown in formula (I) above). In certain embodiments, linker subunit L 2However, if present, the first drug or active agent W 1 It is coupled to the linker subunit L. In a particular embodiment, the linker subunit L 3 However, if present, the first drug or active agent W 1 It is coupled to the linker subunit L. In a particular embodiment, the linker subunit L 4 However, if present, the first drug or active agent W 1 It is coupled to the linker subunit L. In a particular embodiment, the linker subunit L 5 However, if present, the first drug or active agent W 1 It is coupled to the linker subunit L. In a particular embodiment, the linker subunit L 6 However, if present, the first drug or active agent W 1 It is connected.

[0346] First Linker L A In this, any convenient linker subunit may be used. The linker subunit of interest includes, but is not limited to, units of polymers (e.g., polyethylene glycol, polyethylene and polyacrylates, amino acid residues(s), carbohydrate polymers or carbohydrates and their derivatives, polynucleotides, alkyl groups, aryl groups, heterocyclic groups, combinations thereof, and substituted versions thereof). In some embodiments, L 1 , L 2 , L 3 , L 4 , L 5 and L 6 Each of (if present) comprises one or more groups independently selected from polyethylene glycol, modified polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, and diamines (e.g., linking groups including alkylenediamines).

[0347] In some embodiments, L 1 (If present) includes polyethylene glycol, modified polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, or diamines. In some embodiments, L 1It contains polyethylene glycol. In some embodiments, L 1 It contains modified polyethylene glycol. In some embodiments, L 1 It contains an amino acid residue. In some embodiments, L 1 This includes an alkyl group or a substituted alkyl group. In some embodiments, L 1 This includes an aryl group or a substituted aryl group. In some embodiments, L 1 It contains a diamine (for example, a linking group including alkylenediamine).

[0348] In some embodiments, L 2 (If present) includes polyethylene glycol, modified polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, or diamines. In some embodiments, L 2 It contains polyethylene glycol. In some embodiments, L 2 It contains modified polyethylene glycol. In some embodiments, L 2 It contains an amino acid residue. In some embodiments, L 2 This includes an alkyl group or a substituted alkyl group. In some embodiments, L 2 This includes an aryl group or a substituted aryl group. In some embodiments, L 2 It contains a diamine (for example, a linking group including alkylenediamine).

[0349] In some embodiments, L 3 (If present) includes polyethylene glycol, modified polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, or diamines. In some embodiments, L 3 It contains polyethylene glycol. In some embodiments, L 3 It contains modified polyethylene glycol. In some embodiments, L 3 It contains an amino acid residue. In some embodiments, L 3This includes an alkyl group or a substituted alkyl group. In some embodiments, L 3 This includes an aryl group or a substituted aryl group. In some embodiments, L 3 It contains a diamine (for example, a linking group including alkylenediamine).

[0350] In some embodiments, L 4 (If present) includes polyethylene glycol, modified polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, or diamines. In some embodiments, L 4 It contains polyethylene glycol. In some embodiments, L 4 It contains modified polyethylene glycol. In some embodiments, L 4 It contains an amino acid residue. In some embodiments, L 4 This includes an alkyl group or a substituted alkyl group. In some embodiments, L 4 This includes an aryl group or a substituted aryl group. In some embodiments, L 4 It contains a diamine (for example, a linking group including alkylenediamine).

[0351] In some embodiments, L 5 (If present) includes polyethylene glycol, modified polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, or diamines. In some embodiments, L 5 It contains polyethylene glycol. In some embodiments, L 5 It contains modified polyethylene glycol. In some embodiments, L 5 It contains an amino acid residue. In some embodiments, L 5 This includes an alkyl group or a substituted alkyl group. In some embodiments, L 5 This includes an aryl group or a substituted aryl group. In some embodiments, L 5 It contains a diamine (for example, a linking group including alkylenediamine).

[0352] In some embodiments, L 6 (If present) includes polyethylene glycol, modified polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, or diamines. In some embodiments, L 6 It contains polyethylene glycol. In some embodiments, L 6 It contains modified polyethylene glycol. In some embodiments, L 6 It contains an amino acid residue. In some embodiments, L 6 This includes an alkyl group or a substituted alkyl group. In some embodiments, L 6 This includes an aryl group or a substituted aryl group. In some embodiments, L 6 It contains a diamine (for example, a linking group including alkylenediamine).

[0353] In some embodiments, L A teeth, -(L 1 ) a -(L 2 ) b -(L 3 ) c -(L 4 ) d -(L 5 ) e -(L 6 ) f - (In the formula, -(L 1 ) a - is, - (T 1 -V 1 ) a -and; -(L 2 ) b - is, - (T 2 -V 2 ) b -and; -(L 3 ) c - is, - (T 3 -V 3 ) c -and; -(L 4 ) d- is, - (T 4 -V 4 ) d -and; -(L 5 ) e - is, - (T 5 -V 5 ) e -and; -(L 6 ) f - is, - (T 6 -V 6 ) f -and, T 1 , T 2 , T 3 , T 4 , T 5 and T 6 If present, it is a tether group; V 1 , V 2 , V 3 , V 4 , V 5 , and V 6 If present, these are covalent or linking functional groups; a, b, c, d, e, and f are each independently either 0 or 1, provided that at least one of a, b, c, d, e, and f is 1. It is the first linker, which includes [the specified component].

[0354] In certain embodiments, the sum of a, b, c, d, e, and f is between 1 and 6. In certain embodiments, the sum of a, b, c, d, e, and f is 1. In certain embodiments, the sum of a, b, c, d, e, and f is 2. In certain embodiments, the sum of a, b, c, d, e, and f is 3. In certain embodiments, the sum of a, b, c, d, e, and f is 4. In certain embodiments, the sum of a, b, c, d, e, and f is 5. In certain embodiments, the sum of a, b, c, d, e, and f is 6. In certain embodiments, a, b, c, d, e, and f are each 1. In certain embodiments, a, b, c, d, and e are 1, and f is 0. In certain embodiments, a, b, c, and d are each 1, and e and f are each 0. In certain embodiments, a, b, and c are each 1, and d, e, and f are each 0. In certain embodiments, a and b are each 1, and c, d, e, and f are each 0. In certain embodiments, a is 1, and b, c, d, e, and f are each 0.

[0355] As described above, in a particular embodiment, L 1 It is bonded to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety (for example, as shown in formula (I) above). Therefore, in certain embodiments, T 1 It is bonded to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety (for example, as shown in formula (I) above). In certain embodiments, V 1 L is bound to the first drug or active ingredient. In certain embodiments, L 2 If present, it is bound to the first drug or active agent. Therefore, in certain embodiments, T 2 If present, it is bound to the first drug or active ingredient, or V 2 If present, it is bound to the first drug or active ingredient. In certain embodiments, L 3 If present, it is bound to the first drug or active agent. Therefore, in certain embodiments, T3 If present, it is bound to the first drug or active ingredient, or V 3 If present, it is bound to the first drug or active ingredient. In certain embodiments, L 4 If present, it is bound to the first drug or active agent. Therefore, in certain embodiments, T 4 If present, it is bound to the first drug or active ingredient, or V 4 If present, it is bound to the first drug or active ingredient. In certain embodiments, L 5 If present, it is bound to the first drug or active agent. Therefore, in certain embodiments, T 5 If present, it is bound to the first drug or active ingredient, or V 5 If present, it is bound to the first drug or active ingredient. In certain embodiments, L 6 If present, it is bound to the first drug or active agent. Therefore, in certain embodiments, T 6 If present, it is bound to the first drug or active ingredient, or V 6 If present, it is bound to the first drug or active ingredient.

[0356] In certain embodiments, the conjugate of formula (I) is a second linker L B Includes the second linker, L B The second linker, L, can be used to conjugate the desired second portion (e.g., a second drug or active agent) to a ROR antibody, e.g., an activatable ROR antibody, via the conjugation portion. B It can be bonded (e.g., covalently) to a conjugation portion (e.g., as described herein). For example, a second linker, L B The hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety can be bound to the second drug. The hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety can be bound to the second linker, L BTherefore, the second drug can be used to conjugate a ROR antibody, such as an activatable ROR antibody.

[0357] For example, as shown in equation (I) above, L B It is bound to Ab via a conjugation moiety, and therefore Ab is linked to the second linker L via a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety. B It is indirectly bound to L. As described above, Ab is a ROR antibody, for example, an activatable ROR antibody, and thereby L B However, it is bound to ROR antibodies, such as activatable ROR antibodies, via a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety, for example, linker L B However, it indirectly binds to ROR antibodies, such as activatable ROR antibodies, via hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moieties.

[0358] Second linker L in the subject conjugate and compound B Any suitable linker can be used. In a particular embodiment, a second linker L B However, it may also contain a group selected from alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acylamino, alkylamide, substituted alkylamide, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, the second linker L B It may contain alkyl or substituted alkyl groups. In certain embodiments, a second linker L B This may contain an alkenyl group or a substituted alkenyl group. In certain embodiments, the second linker L B This may contain an alkynyl group or a substituted alkynyl group. In certain embodiments, the second linker LB It may contain an alkoxy group or a substituted alkoxy group. In certain embodiments, the second linker L B It may contain an amino group or a substituted amino group. In certain embodiments, the second linker L B It may contain a carboxyl group or a carboxyl ester group. In certain embodiments, the second linker L B It may contain an acylamino group. In certain embodiments, the second linker L B This may include an alkylamide group or a substituted alkylamide group. In certain embodiments, the second linker L B This may include an aryl group or a substituted aryl group. In certain embodiments, the second linker L B This may include a heteroaryl group or a substituted heteroaryl group. In certain embodiments, the second linker L B This may include a cycloalkyl or substituted cycloalkyl group. In certain embodiments, a second linker L B This may contain a heterocyclyl group or a substituted heterocyclyl group.

[0359] In certain embodiments, a second linker L B The polymer may include, for example, polyalkylene glycols and their derivatives (e.g., polyethylene glycol, methoxypolyethylene glycol, polyethylene glycol homopolymer, polypropylene glycol homopolymer, copolymers of ethylene glycol and propylene glycol (e.g., homopolymers and copolymers are unsubstituted or have one end substituted with an alkyl group)), polyvinyl alcohol, polyvinyl ethyl, ether, polyvinylpyrrolidone, and combinations thereof. In certain embodiments, the polymer is polyalkylene glycol. In certain embodiments, the polymer is polyethylene glycol. Other linkers are also usable, as shown in the conjugates and compounds described in more detail below.

[0360] In some embodiments, L B The formula is as follows: -(L 7 ) g -(L 8 ) h -(L 9 ) i -(L 10 ) j -(L 11 ) k -(L 12 ) l -(L 13 ) m (In the formula, L 7 , L 8 , L 9 , L 10 , L 11 , L 12 and L 13 Each of these is an independent linker subunit, and each of g, h, i, j, k, l, and m is independently 0 or 1, provided that at least one of g, h, i, j, k, l, and m is 1. It is the second linker described by [author's name].

[0361] In certain embodiments, the sum of g, h, i, j, k, l, and m is between 1 and 7. In certain embodiments, the sum of g, h, i, j, k, l, and m is 1. In certain embodiments, the sum of g, h, i, j, k, l, and m is 2. In certain embodiments, the sum of g, h, i, j, k, l, and m is 3. In certain embodiments, the sum of g, h, i, j, k, l, and m is 4. In certain embodiments, the sum of g, h, i, j, k, l, and m is 5. In certain embodiments, the sum of g, h, i, j, k, l, and m is 6. In certain embodiments, the sum of g, h, i, j, k, l, and m is 7. In certain embodiments, g, h, i, j, k, l, and m are each 1. In certain embodiments, g, h, i, j, k, and l are each 1, and m is 0. In a particular embodiment, g, h, i, j, and k are each 1, and l and m are each 0. In a particular embodiment, g, h, i, and j are each 1, and k, l, and m are each 0. In a particular embodiment, g, h, and i are each 1, and j, k, l, and m are each 0. In a particular embodiment, g and h are each 1, and i, j, k, l, and m are each 0. In a particular embodiment, g is 1, and h, i, j, k, l, and m are each 0. In a particular embodiment, g, h, i, j, k, l, and m are each 0.

[0362] In a particular embodiment, linker subunit L 7 However, it is bonded to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety (for example, as shown in formula (I) above). In certain embodiments, linker subunit L 8 However, if present, a second drug or active agent W 2 It is coupled to the linker subunit L. In a particular embodiment, the linker subunit L 9 However, if present, a second drug or active agent W 2 It is coupled to the linker subunit L. In a particular embodiment, the linker subunit L 10 However, if present, a second drug or active agent W 2It is coupled to the linker subunit L. In a particular embodiment, the linker subunit L 11 However, if present, a second drug or active agent W 2 It is coupled to the linker subunit L. In a particular embodiment, the linker subunit L 12 However, if present, a second drug or active agent W 2 It is coupled to the linker subunit L. In a particular embodiment, the linker subunit L 13 However, if present, a second drug or active agent W 2 It is connected.

[0363] Any suitable linker subunit can be used as the second linker L B It can be used for the following. The linker subunit of interest includes, but is not limited to, units of polymers (e.g., polyethylene glycol, polyethylene and polyacrylates, amino acid residues(s), carbohydrate polymers or carbohydrates and their derivatives, polynucleotides, alkyl groups, aryl groups, heterocyclic groups, combinations thereof, and substituted versions thereof). In some embodiments, L 7 , L 8 , L 9 , L 10 , L 11 , L 12 and L 13 Each of (if present) comprises one or more groups independently selected from polyethylene glycol, modified polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, and diamines (e.g., linking groups including alkylenediamines).

[0364] In some embodiments, L 7 (If present) includes polyethylene glycol, modified polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, or diamines. In some embodiments, L 7 It contains polyethylene glycol. In some embodiments, L 7 It contains modified polyethylene glycol. In some embodiments, L 7It contains an amino acid residue. In some embodiments, L 7 This includes an alkyl group or a substituted alkyl group. In some embodiments, L 7 This includes an aryl group or a substituted aryl group. In some embodiments, L 7 It contains a diamine (for example, a linking group including alkylenediamine).

[0365] In some embodiments, L 8 (If present) includes polyethylene glycol, modified polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, or diamines. In some embodiments, L 8 It contains polyethylene glycol. In some embodiments, L 8 It contains modified polyethylene glycol. In some embodiments, L 8 It contains an amino acid residue. In some embodiments, L 8 This includes an alkyl group or a substituted alkyl group. In some embodiments, L 8 This includes an aryl group or a substituted aryl group. In some embodiments, L 8 It contains a diamine (for example, a linking group including alkylenediamine).

[0366] In some embodiments, L 9 (If present) includes polyethylene glycol, modified polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, or diamines. In some embodiments, L 9 It contains polyethylene glycol. In some embodiments, L 9 It contains modified polyethylene glycol. In some embodiments, L 9 It contains an amino acid residue. In some embodiments, L 9 This includes an alkyl group or a substituted alkyl group. In some embodiments, L 9 This includes an aryl group or a substituted aryl group. In some embodiments, L 9It contains a diamine (for example, a linking group including alkylenediamine).

[0367] In some embodiments, L 10 (If present) includes polyethylene glycol, modified polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, or diamines. In some embodiments, L 10 It contains polyethylene glycol. In some embodiments, L 10 It contains modified polyethylene glycol. In some embodiments, L 10 It contains an amino acid residue. In some embodiments, L 10 This includes an alkyl group or a substituted alkyl group. In some embodiments, L 10 This includes an aryl group or a substituted aryl group. In some embodiments, L 10 It contains a diamine (for example, a linking group including alkylenediamine).

[0368] In some embodiments, L 11 (If present) includes polyethylene glycol, modified polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, or diamines. In some embodiments, L 11 It contains polyethylene glycol. In some embodiments, L 11 It contains modified polyethylene glycol. In some embodiments, L 11 It contains an amino acid residue. In some embodiments, L 11 This includes an alkyl group or a substituted alkyl group. In some embodiments, L 11 This includes an aryl group or a substituted aryl group. In some embodiments, L 11 It contains a diamine (for example, a linking group including alkylenediamine).

[0369] In some embodiments, L 12(If present) includes polyethylene glycol, modified polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, or diamines. In some embodiments, L 12 It contains polyethylene glycol. In some embodiments, L 12 It contains modified polyethylene glycol. In some embodiments, L 12 It contains an amino acid residue. In some embodiments, L 12 This includes an alkyl group or a substituted alkyl group. In some embodiments, L 12 This includes an aryl group or a substituted aryl group. In some embodiments, L 12 It contains a diamine (for example, a linking group including alkylenediamine).

[0370] In some embodiments, L 13 (If present) includes polyethylene glycol, modified polyethylene glycol, amino acid residues, alkyl groups, substituted alkyl groups, aryl groups, substituted aryl groups, or diamines. In some embodiments, L 13 It contains polyethylene glycol. In some embodiments, L 13 It contains modified polyethylene glycol. In some embodiments, L 13 It contains an amino acid residue. In some embodiments, L 13 This includes an alkyl group or a substituted alkyl group. In some embodiments, L 13 This includes an aryl group or a substituted aryl group. In some embodiments, L 13 It contains a diamine (for example, a linking group including alkylenediamine).

[0371] In some embodiments, L B teeth, -(L 7 ) g -(L 8 ) h -(L 9 ) i -(L 10 ) j -(L 11 )k -(L 12 ) l -(L 13 ) m - (In the formula, -(L 7 ) g - is, - (T 7 -V 7 ) g -and; -(L 8 ) h - is, - (T 8 -V 8 ) h -and; -(L 9 ) i - is, - (T 9 -V 9 ) i -and; -(L 10 ) j - is, - (T 10 -V 10 ) j -and; -(L 11 ) k - is, - (T 11 -V 11 ) k -and; -(L 12 ) l - is, - (T 12 -V 12 ) l -and; -(L 13 ) m - is, - (T 13 -V 13 ) m -and, T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 If present, it is a tether group; V 7 , V 8 , V 9 , V 10 , V 11 , V 12, and V 13 If present, these are covalent or linking functional groups; g, h, i, j, k, l, and m are each independently either 0 or 1, provided that at least one of g, h, i, j, k, l, and m is 1. It is the second linker, which includes [the specified component].

[0372] In certain embodiments, the sum of g, h, i, j, k, l, and m is between 1 and 7. In certain embodiments, the sum of g, h, i, j, k, l, and m is 1. In certain embodiments, the sum of g, h, i, j, k, l, and m is 2. In certain embodiments, the sum of g, h, i, j, k, l, and m is 3. In certain embodiments, the sum of g, h, i, j, k, l, and m is 4. In certain embodiments, the sum of g, h, i, j, k, l, and m is 5. In certain embodiments, the sum of g, h, i, j, k, l, and m is 6. In certain embodiments, the sum of g, h, i, j, k, l, and m is 7. In certain embodiments, g, h, i, j, k, l, and m are each 1. In certain embodiments, g, h, i, j, k, and l are each 1, and m is 0. In a particular embodiment, g, h, i, j, and k are each 1, and l and m are each 0. In a particular embodiment, g, h, i, and j are each 1, and k, l, and m are each 0. In a particular embodiment, g, h, and i are each 1, and j, k, l, and m are each 0. In a particular embodiment, g and h are each 1, and i, j, k, l, and m are each 0. In a particular embodiment, g is 1, and h, i, j, k, l, and m are each 0. In a particular embodiment, g, h, i, j, k, l, and m are each 0.

[0373] As described above, in a particular embodiment, L 7 It is bonded to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety (for example, as shown in formula (I) above). Therefore, in certain embodiments, T 7It is bonded to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety (for example, as shown in formula (I) above). In certain embodiments, V 7 However, it is bound to a second drug or active ingredient. In certain embodiments, L 8 However, if present, it is bound to the second drug or active agent. Therefore, in certain embodiments, T 8 However, if present, it is bound to the second drug or active ingredient, or V 8 However, if present, it is bound to the second drug or active agent. In certain embodiments, L 9 However, if present, it is bound to the second drug or active agent. Therefore, in certain embodiments, T 9 However, if present, it is bound to the second drug or active ingredient, or V 9 However, if present, it is bound to the second drug or active agent. In certain embodiments, L 10 However, if present, it is bound to the second drug or active agent. Therefore, in certain embodiments, T 10 However, if present, it is bound to the second drug or active ingredient, or V 10 However, if present, it is bound to the second drug or active agent. In certain embodiments, L 11 However, if present, it is bound to the second drug or active agent. Therefore, in certain embodiments, T 11 However, if present, it is bound to the second drug or active ingredient, or V 11 However, if present, it is bound to the second drug or active agent. In certain embodiments, L 12 However, if present, it is bound to the second drug or active agent. Therefore, in certain embodiments, T 12 However, if present, it is bound to the second drug or active ingredient, or V 12 However, if present, it is bound to the second drug or active agent. In certain embodiments, L 13 However, if present, it is bound to the second drug or active agent. Therefore, in certain embodiments, T 13However, if present, it is bound to the second drug or active ingredient, or V 13 However, if present, it is bound to the second drug or active ingredient.

[0374] Tether group T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 Regarding this, any suitable tether group can be used as the linker of the subject. In some embodiments, T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 However, each 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 OH) x- comprises one or more groups independently selected from 4-amino-piperidine (4AP), meta-aminobenzyloxy (MABO), meta-aminobenzyloxycarbonyl (MABC), para-aminobenzyloxy (PABO), para-aminobenzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-aminobenzylamino (PABA), para-aminophenyl (PAP), para-hydroxyphenyl (PHP), acetal groups, hydrazines, disulfides, and esters, where each w is an integer from 1 to 20, each n is an integer from 1 to 30, each p is an integer from 1 to 20, and each x is an integer from 1 to 12.

[0375] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and / or T 13 ) but (C1~C 12 ) alkyl or substituted (C1~C 12 ) Contains alkyl. In certain embodiments, (C1~C 12 )The alkyl group is a linear or branched alkyl group containing 1 to 12 carbon atoms (for example, 1 to 10 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, or 1 to 3 carbon atoms). In some cases, (C1-C 12 ) Alkyl is alkyl or substituted alkyl (e.g., C1~C 12 Alkyl, or C1-C 10 It may be alkyl, or C1-C6 alkyl, or C1-C3 alkyl. In some cases, (C1-C 12 ) Alkyl is a C2 alkyl. For example, (C1~C 12 ) Alkyl is alkylene or substituted alkylene (e.g., C1~C 12 Alkylene, or C1-C 10It may be an alkylene, or a C1-C6 alkylene, or a C1-C3 alkylene. In some cases, (C1-C 12 )alkyl is C1-alkylene (e.g., CH2). In some cases, (C1-C 12 )The alkyl group is C2-alkylene (e.g., CH2CH2). In some cases, (C1-C 12 The alkyl group is a C3-alkylene (e.g., CH2CH2CH2).

[0376] In certain embodiments, substitution (C1~C 12 )The alkyl is a linear or branched substituted alkyl group containing 1 to 12 carbon atoms (e.g., 1 to 10 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, or 1 to 3 carbon atoms). In some cases, the substitution (C1-C 12 )alkyl is a substituted alkyl (for example, substituted C1~C 12 Alkyl or substituted C1-C 10 It may be alkyl, or substituted C1-C6 alkyl, or substituted C1-C3 alkyl. In some cases, substitution (C1-C 12 )alkyl is a substituted C2 alkyl. For example, substituted (C1~C 12 )alkyl is a substituted alkylene (for example, substituted C1~C 12 Alkylene, or substituted C1-C 10 It can be an alkylene, or a substituted C1-C6 alkylene, or a substituted C1-C3 alkylene. In some cases, substitution (C1-C 12 )alkyl is a substituted C1-alkylene (e.g., a C1-alkylene substituted with -SO3H). In some cases, substitution (C1-C 12 )alkyl is a substituted C2 alkylene. In some cases, substituted (C1-C 12 )alkyl is a substituted C3 alkylene. For example, substituted (C1-C 12 ) Alkyl, C1-C 12 Alkylene (e.g., (PEG) as described herein) kC3-alkylene or C5-alkylene substituted with a group (e.g., -CONH(PEG)) k For example, -CONH(PEG)3 or -CONH(PEG)5; or -NHCO(PEG) k (For example, even if it contains -NHCO(PEG)7, C1-C substituted with a -CONHCH2CH2SO3H group) 12 Even if it contains alkylene (e.g., C3-alkylene), C1-C substituted with an -NHCOCH2SO3H group 12 It may also contain alkylenes (e.g., C5-alkylenes).

[0377] In some embodiments, substitution (C1-C 12 ) Alkyl as described herein (PEG) t Base (for example, -NHCO(PEG)) t (In the formula, (PEG) t teeth, [ka] And, [ka] ∫ indicates the bond site to the carbonyl group of -NHCO-, and t is an integer), for example, C1-C substituted with -NHCO(CH2CH2O)3CH3, -NHCO(CH2CH2O)5CH3, or -NHCO(CH2CH2O)8CH3. 12 It may also contain alkylenes (e.g., C3-alkylene or C5-alkylene).

[0378] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and / or T 13) includes aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, or substituted heterocyclyl. In some cases, a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and / or T 13 ) contains an aryl or substituted aryl. For example, the aryl can be a phenyl. In some cases, the substituted aryl is a substituted phenyl. A substituted phenyl is (C1-C 12 ) alkyl, substituted (C1-C 12 The substituents can be substituted with one or more substituents selected from alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In some examples, the substituted aryl is a substituted phenyl, and the substituents include cleavable moieties as described herein (e.g., enzymatically cleavable moieties, e.g., glycosides or glycoside derivatives).

[0379] In some cases, a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and / or T 13 ) includes a heteroaryl or substituted heteroaryl, for example, a triazolyl (e.g., 1,2,3-triazolyl). In some cases, a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and / or T 13 ) includes a cycloalkyl or substituted cycloalkyl group. In some cases, a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and / or T 13 ) comprises a heterocyclyl or a substituted heterocyclyl. In some examples, the substituents on a substituted heteroaryl, substituted cycloalkyl, or substituted heterocyclyl comprise the cleavable moieties described herein (e.g., enzymatically cleavable moieties, e.g., glycosides or glycoside derivatives).

[0380] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and / or T 13 ) comprises an ethylenediamine (EDA) moiety (e.g., an EDA-containing tether group). In certain embodiments, (EDA) wHowever, it includes one or more EDA moieties, for example, where w is an integer from 1 to 50, e.g., 1 to 40, 1 to 30, 1 to 20, 1 to 12, or 1 to 6, e.g., 1, 2, 3, 4, 5, or 6. The bonded ethylenediamine (EDA) moieties may optionally be substituted at one or more convenient positions with any convenient substituent (e.g., alkyl, substituted alkyl, acyl, substituted acyl, aryl, or substituted aryl). In certain embodiments, the EDA moiety is described with the following structure: [ka] (In the formula, y is an integer from 1 to 6, r is 0 or 1, and each R 12 (However, independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, 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 certain embodiments, y is 2 and r is 0. In certain embodiments, y is 2 and r is 1. In certain embodiments, each R 12 However, these are independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. In certain embodiments, any two adjacent R of EDA 12 The groups may be cyclically bonded, for example, to form a piperazinyl ring. In certain embodiments, y is 1 and two adjacent R 12 The group is an alkyl group, which is cyclically bonded to form a piperazinyl ring. In certain embodiments, y is 1, and adjacent R 12The group is selected from hydrogen, alkyl (e.g., methyl), and substituted alkyl (e.g., lower alkyl-OH, e.g., ethyl-OH or propyl-OH).

[0381] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and / or T 13 ) comprises a 4-amino-piperidine (4AP) moiety (also referred herein as piperidine-4-amino, P4A). The 4AP moiety may optionally be substituted at one or more convenient positions with any convenient substituent (e.g., alkyl, substituted alkyl, polyethylene glycol moiety, acyl, substituted acyl, aryl, or substituted aryl). In certain embodiments, the 4AP moiety is described with the following structure: [ka] (In the formula, R 12 (Selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety (e.g., polyethylene glycol or modified polyethylene glycol), alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl). In certain embodiments, R 12 However, this is the polyethylene glycol portion. In a particular embodiment, R 12 However, it is carboxy-modified polyethylene glycol.

[0382] In a particular embodiment, R 12 However, formula: (PEG) k It includes the polyethylene glycol portion described in, which can be represented by the following structure: [ka] (wherein k is an integer from 1 to 20, e.g., 1 to 18, 1 to 16, 1 to 14, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 4, 1 to 2, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20). In some cases, k is 2. In certain embodiments, R 17 R is selected from OH, COOH, OR, or COOR, and R is selected from alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R 17 However, it is COOH. In a particular embodiment, R 17 However, it is OH. In a particular embodiment, R 17 However, it is OCH3.

[0383] In certain embodiments, (PEG) k (PEG) has the following structure t is: [ka] (In the formula, t is an integer between 2 and 10). In a particular embodiment, t is 8.

[0384] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11, T 12 , and / or T 13 ) but (PEG) n (PEG) n However, it is a polyethylene glycol or modified polyethylene glycol bonded unit. In certain embodiments, (PEG) n However, it is written in the following structure: [ka] (In the formula, n is an integer between 1 and 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, 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.

[0385] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and / or T 13 ) but (AA) p It includes, where AA is an amino acid residue. Any suitable amino acid may be used. The amino acid of interest includes, but is not limited to, L-amino acids and D-amino acids, natural amino acids (e.g., any of the 20 major α-amino acids and β-alanine), and unnatural amino acids (e.g., amino acid analogs), such as unnatural α-amino acids or unnatural β-amino acids. In certain embodiments, p is an integer from 1 to 50, e.g., 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 certain embodiments, p is 1. In certain embodiments, p is 2.

[0386] In further embodiments, (AA) p It contains a valine-alanine dipeptide.

[0387] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and / or T 13 ) includes amino acid analogs. Amino acid analogs include compounds that are structurally and / or overall similar in shape to one or more amino acids commonly found in natural proteins (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, or Tyr or Y). Amino acid analogs also include natural amino acids with modified side chains or backbones. Amino acid analogs also include amino acid analogs that have the same stereochemistry as naturally occurring D- and L-type amino acid analogs. In some cases, amino acid analogs share the backbone structure and / or side chain structure of one or more natural amino acids, but the difference(s) is one or more modification groups within the molecule. Such modifications may include, but are not limited to, the substitution of an atom (e.g., N) for a related atom (e.g., S), the addition of a group (e.g., methyl, hydroxyl, etc.) or an atom (e.g., Cl, Br, etc.), the deletion of a group, the substitution of a covalent bond (e.g., a single bond to a double bond), or combinations thereof. For example, an amino acid analog may include α-hydroxy acids and α-amino acids. Examples of amino acid analogs include, but are not limited to, sulfoalanine.

[0388] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and / or T 13 ) is formula - (CR 13 OH) x The portion indicated by - is such that x is either 0 or x is an integer between 1 and 50, e.g., 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, or 12. In certain embodiments, x is 1. In certain embodiments, x is 2. In certain embodiments, R 13 The following are selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R 13 is hydrogen. In a particular embodiment, R 13 is alkyl or substituted alkyl (e.g., C 1-6 Alkyl or C 1-6 Substituting alkyl, or C 1-4 Alkyl or C 1-4 Substituting alkyl, or C 1-3 Alkyl or C 1-3 It is a substituted alkyl. In certain embodiments, R 13 is an alkenyl or substituted alkenyl (e.g., C 2-6 Alkenil or C 2-6 Substituting alkenyl, or C2-4 Alkenil or C 2-4 Substituting alkenyl, or C 2-3 Alkenil or C 2-3 It is a substituted alkenyl. In certain embodiments, R 13 is an alkynyl or substituted alkynyl. In certain embodiments, R 13 is an alkoxy or substituted alkoxy. In certain embodiments, R 13 is an amino acid or a substituted amino acid. In certain embodiments, R 13 is a carboxyl or carboxyl ester. In certain embodiments, R 13 is an acyl or acyloxy. In certain embodiments, R 13 is acylamino or aminoacyl. In certain embodiments, R 13 is an alkylamide or a substituted alkylamide. In certain embodiments, R 13 is a sulfonyl. In certain embodiments, R 13 is a thioalkoxy or substituted thioalkoxy. In certain embodiments, R 13 is an aryl or substituted aryl (for example, C 5-8 Aryl or C 5-8 Substituting aryls (for example, C5 aryl or C5-substituted aryl, or C6 aryl or C6-substituted aryl). In certain embodiments, R 13 This refers to heteroaryl or substituted heteroaryl (e.g., C 5-8 heteroaryl or C 5-8 A substituted heteroaryl, for example, a C5 heteroaryl or a C5-substituted heteroaryl, or a C6 heteroaryl or a C6-substituted heteroaryl. In certain embodiments, R 13 This refers to cycloalkyl or substituted cycloalkyl (e.g., C 3-8 Cycloalkyl or C 3-8 Substitutive cycloalkyls, for example, C 3-6 Cycloalkyl or C 3-6 Substituted cycloalkyl, or C 3-5 Cycloalkyl or C 3-5It is a substituted cycloalkyl. In certain embodiments, R 13 This refers to heterocyclines or substituted heterocyclines (e.g., C 3-8 Heterocycline or C 3-8 Substituting heterocyclyls, e.g., C 3-6 Heterocycline or C 3-6 Substituted heterocyclyl, or C 3-5 Heterocycline or C 3-5 It is a substituted heterocyclyl.

[0389] In a particular embodiment, R 13 However, it is selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. In these embodiments, alkyl, substituted alkyl, aryl, and substituted aryl are R 13 The above is true.

[0390] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and / or T 13 The tether group comprises an acetal group, a disulfide, a hydrazine, or an ester. In some embodiments, the tether group comprises an acetal group. In some embodiments, the tether group comprises a hydrazine. In some embodiments, the tether group comprises a disulfide. In some embodiments, the tether group comprises an ester.

[0391] In certain embodiments, a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T11 , T 12 , and / or T 13 This includes meta-aminobenzyloxy (MABO), meta-aminobenzyloxycarbonyl (MABC), para-aminobenzyloxy (PABO), para-aminobenzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-aminobenzylamino (PABA), para-aminophenyl (PAP), or para-hydroxyphenyl (PHP).

[0392] In some embodiments, the tether group includes a MABO group described in the following structure: [ka]

[0393] In some embodiments, the tether group includes a MABC group described in the following structure: [ka]

[0394] In some embodiments, the tether group includes a PABO group described in the following structure: [ka] .

[0395] In some embodiments, the tether group includes a PABC group described in the following structure: [ka] .

[0396] In some embodiments, the tether group includes a PAB group described in the following structure: [ka] .

[0397] In some embodiments, the tether group includes a PABA group described in the following structure: [ka] .

[0398] In some embodiments, the tether group includes a PAP group described in the following structure: [ka] .

[0399] In some embodiments, the tether group includes a PHP group described in the following structure: [ka] .

[0400] In a specific method of operation, each R 14 These are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

[0401] In a particular embodiment, R 14 is hydrogen. In a particular embodiment, each R 14 is hydrogen. In a particular embodiment, R 14 is alkyl or substituted alkyl (e.g., C 1-6 Alkyl or C 1-6 Substituting alkyl, or C 1-4 Alkyl or C 1-4 Substituting alkyl, or C 1-3 Alkyl or C 1-3 It is a substituted alkyl. In certain embodiments, R14 is an alkenyl or substituted alkenyl (e.g., C 2-6 Alkenil or C 2-6 Substituting alkenyl, or C 2-4 Alkenil or C 2-4 Substituting alkenyl, or C 2-3 Alkenil or C 2-3 It is a substituted alkenyl. In certain embodiments, R 14 is an alkynyl or substituted alkynyl. In certain embodiments, R 14 is an alkoxy or substituted alkoxy. In certain embodiments, R 14 is an amino acid or a substituted amino acid. In certain embodiments, R 14 is a carboxyl or carboxyl ester. In certain embodiments, R 14 is an acyl or acyloxy. In certain embodiments, R 14 is acylamino or aminoacyl. In certain embodiments, R 14 is an alkylamide or a substituted alkylamide. In certain embodiments, R 14 is a sulfonyl. In certain embodiments, R 14 is a thioalkoxy or substituted thioalkoxy. In certain embodiments, R 14 is an aryl or substituted aryl (for example, C 5-8 Aryl or C 5-8 Substituting aryls (for example, C5 aryl or C5-substituted aryl, or C6 aryl or C6-substituted aryl). In certain embodiments, R 14 This refers to heteroaryl or substituted heteroaryl (e.g., C 5-8 heteroaryl or C 5-8 A substituted heteroaryl, for example, a C5 heteroaryl or a C5-substituted heteroaryl, or a C6 heteroaryl or a C6-substituted heteroaryl. In certain embodiments, R 14 This refers to cycloalkyl or substituted cycloalkyl (e.g., C 3-8 Cycloalkyl or C 3-8 Substitutive cycloalkyls, for example, C 3-6Cycloalkyl or C 3-6 Substituted cycloalkyl, or C 3-5 Cycloalkyl or C 3-5 It is a substituted cycloalkyl. In certain embodiments, R 14 This refers to heterocyclines or substituted heterocyclines (e.g., C 3-8 Heterocycline or C 3-8 Substituting heterocyclyls, e.g., C 3-6 Heterocycline or C 3-6 Substituted heterocyclyl, or C 3-5 Heterocycline or C 3-5 It is a substituted heterocyclyl.

[0402] In some embodiments of the MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP tether structures shown above, the phenyl ring may be substituted with one or more additional groups selected from halogens, alkyls, substituted alkyls, alkenyls, substituted alkenyls, alkynyls, substituted alkynyls, alkoxys, substituted alkoxys, aminos, substituted aminos, carboxyls, carboxyl esters, acyls, acyloxys, acylaminos, aminoacyls, alkylamides, substituted alkylamides, sulfonyls, thioalkoxys, substituted thioalkoxys, aryls, substituted aryls, heteroaryls, substituted heteroaryls, cycloalkyls, substituted cycloalkyls, heterocyclyls, and substituted heterocyclyls.

[0403] In certain embodiments, the tether group T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and / or T 13 One or more of these are optionally substituted with a glycoside or a glycoside derivative. For example, in some cases, T 1 , T 2 , T3 , T 4 , T 5 and T 6 Each of these is optionally substituted with a glycoside. In some cases, T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 Each of these is optionally substituted with a glycoside. In certain embodiments, the glycoside or glycoside derivative is selected from glucuronide, galactoside, glucoside, mannoside, fucoside, O-GlcNAc, and O-GalNAc.

[0404] In certain embodiments, the MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP tether structures shown above may be substituted with one or more additional groups selected from glycosides and glycoside derivatives. For example, in some embodiments of the MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP tether structures shown above, the phenyl ring may be substituted with one or more additional groups selected from glycosides and glycoside derivatives. In certain embodiments, the glycoside or glycoside derivative is selected from glucuronide, galactoside, glucoside, mannoside, fucoside, O-GlcNAc, and O-GalNAc. In some embodiments, PABC is substituted with a glycoside, for example, the hydrogens of PABC are substituted with glycosides (e.g., glucuronide, galactoside, glucoside, mannoside, fucoside, O-GlcNAc, and O-GalNAc).

[0405] For example, in some embodiments, the glycoside or glycoside derivative is selected from the following structures: [ka]

[0406] Linking functional group V 1 , V 2 , V 3 , V4 , V 5 , V 6 , V 7 , V 8 , V 9 , V 10 , V 11 , V 12 , and V 13 With regard to this, any convenient linking functional group can be used in the linker of the subject. The linking functional groups of interest include, but are not limited to, amino, carbonyl, amide, oxycarbonyl, carboxy, sulfonyl, sulfoxide, sulfonylamino, aminosulfonyl, thio, oxy, phospho, phosphoramidate, thiophosphorimidate, etc. In some embodiments, V 1 , V 2 , V 3 , V 4 , V 5 , V 6 , V 7 , V 8 , V 9 , V 10 , V 11 , V 12 , and V 13 These are, independently, covalent, -CO-, and -NR bonds. 15 -, -NR 15 (CH2) q -, -NR 15 (C6H4)-, -CONR 15 -, -NR 15 CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR 15 -, -NR 15 Selected from SO2- and -P(O)OH-, where q is an integer from 1 to 6. In certain embodiments, q is an integer from 1 to 6 (e.g., 1, 2, 3, 4, 5, or 6). In certain embodiments, q is 1. In certain embodiments, q is 2. In certain embodiments, q is 3. In certain embodiments, q is 4. In certain embodiments, q is 5. In certain embodiments, q is 6.

[0407] Several embodiments, each R 15These are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, aminoacyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

[0408] In a particular embodiment, R 15 is hydrogen. In a particular embodiment, each R 15 is hydrogen. In a particular embodiment, R 15 is alkyl or substituted alkyl (e.g., C 1-6 Alkyl or C 1-6 Substituting alkyl, or C 1-4 Alkyl or C 1-4 Substituting alkyl, or C 1-3 Alkyl or C 1-3 It is a substituted alkyl. In certain embodiments, R 15 is an alkenyl or substituted alkenyl (e.g., C 2-6 Alkenil or C 2-6 Substituting alkenyl, or C 2-4 Alkenil or C 2-4 Substituting alkenyl, or C 2-3 Alkenil or C 2-3 It is a substituted alkenyl. In certain embodiments, R 15 is an alkynyl or substituted alkynyl. In certain embodiments, R 15 is an alkoxy or substituted alkoxy. In certain embodiments, R 15 is an amino acid or a substituted amino acid. In certain embodiments, R 15 is a carboxyl or carboxyl ester. In certain embodiments, R 15 is an acyl or acyloxy. In certain embodiments, R 15 is acylamino or aminoacyl. In certain embodiments, R 15is an alkylamide or a substituted alkylamide. In certain embodiments, R 15 is a sulfonyl. In certain embodiments, R 15 is a thioalkoxy or substituted thioalkoxy. In certain embodiments, R 15 is an aryl or substituted aryl (for example, C 5-8 Aryl or C 5-8 Substituting aryls (for example, C5 aryl or C5-substituted aryl, or C6 aryl or C6-substituted aryl). In certain embodiments, R 15 This refers to heteroaryl or substituted heteroaryl (e.g., C 5-8 heteroaryl or C 5-8 A substituted heteroaryl, for example, a C5 heteroaryl or a C5-substituted heteroaryl, or a C6 heteroaryl or a C6-substituted heteroaryl. In certain embodiments, R 15 This refers to cycloalkyl or substituted cycloalkyl (e.g., C 3-8 Cycloalkyl or C 3-8 Substitutive cycloalkyls, for example, C 3-6 Cycloalkyl or C 3-6 Substituted cycloalkyl, or C 3-5 Cycloalkyl or C 3-5 It is a substituted cycloalkyl. In certain embodiments, R 15 This refers to heterocyclines or substituted heterocyclines (e.g., C 3-8 Heterocycline or C 3-8 Substituting heterocyclyls, e.g., C 3-6 Heterocycline or C 3-6 Substituted heterocyclyl, or C 3-5 Heterocycline or C 3-5 It is a substituted heterocyclyl.

[0409] In a specific method of operation, each R 15R is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, 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, heterocyclyl, and substituted heterocyclyl is R 15 The above is true.

[0410] As described above, in some embodiments, L A is, -(T 1 -V 1 ) a -(T 2 -V 2 ) b -(T 3 -V 3 ) c -(T 4 -V 4 ) d -(T 5 -V 5 ) e -(T 6 -V 6 ) f -A first linker comprising (wherein a, b, c, d, e, and f are independently either 0 or 1, except that at least one of a, b, c, d, e, and f is 1).

[0411] In some embodiments, the first linker L A In: T 1 (C1-C 12 )alkyl and substituted (C1-C 12 ) Selected from alkyl groups; T 2 , T 3 , T 4 , T 5 and T 6(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 OH) x -, independently selected from 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, acetal group, disulfide, hydrazine, and ester; V 1 , V 2 , V 3 , V 4 , V 5 , and V 6 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-, -SO2NR 15 -, -NR 15 The molecules are independently selected from SO2- and -P(O)OH-, where q is an integer between 1 and 6; (PEG) n teeth, [ka] And n is an integer from 1 to 30; The EDA has the following structure: [ka] The ethylenediamine moiety has , where y is an integer from 1 to 6, and r is 0 or 1; 4-amino-piperidine (4AP) is, [ka] and; AA is an amino acid residue, and p is an integer between 1 and 20; Each R 12 R is independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, and any two adjacent R 12 The groups can be linked in a cyclic manner to form a piperazinyl ring; Each R 13 This is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl; Each R 15 This is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

[0412] In some embodiments, L A teeth, -(T 1 -V 1 ) a -(T 2 -V 2 ) b -(T 3 -V 3 ) c -(T 4 -V 4 ) d -(T 5 -V 5 ) e -(T 6 -V 6 ) f - (In the formula, a, b, c, d, e, and f are each independently either 0 or 1, provided that at least one of a, b, c, d, e, and f is 1; T 1 , T 2 , T 3 , T 4 , T 5 and T 6 is a covalent bond, (C1-C12 ) 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 OH) x -, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxyphenyl (PHP), acetal group, hydrazine, disulfide, and ester, each independently selected from -, 4-amino-piperidine (4AP), meta-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxyphenyl (PHP), acetal group, hydrazine, disulfide, and ester, where EDA is the ethylenediamine moiety, PEG is polyethylene glycol, AA is an amino acid residue or amino acid analog, each w is an integer from 1 to 20, each n is an integer from 1 to 30, each p is an integer from 1 to 20, and each x is an integer from 1 to 12; V 1 , V 2 , V 3 , V 4 , V 5 , and V 6 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-, -SO2NR 15 -, -NR 15 The groups SO2- and -P(O)OH- are independently selected, and each q is an integer from 1 to 6; Each R 13 This is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl; Each R15 (This is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.) Includes.

[0413] L A In some embodiments, T 1 (C1-C 12 )alkyl and substituted (C1-C 12 ) Selected from alkyl groups; T 2 , T 3 , T 4 , T 5 and T 6 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 OH) x -, independently selected from 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, acetal group, hydrazine, and ester; V 1 , V 2 , V 3 , V 4 , V 5 , and V 6 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-, -SO2NR15 -, -NR 15 Independently selected from the group consisting of SO2- and -P(O)OH-; (PEG) n teeth, [ka] And n is an integer from 1 to 30; The EDA has the following structure: [ka] The ethylenediamine moiety has , where y is an integer from 1 to 6, and r is 0 or 1; 4-amino-piperidine (4AP) is, [ka] and; Each R 12 R is independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, and any two adjacent R 12 The groups can be linked in a cyclic manner to form a piperazinyl ring; a, b, c, and d are each 1; (e and f are 0) Includes.

[0414] In some embodiments, T 1 , T 2 , T 3 , T 4 , T 5 and T 6 These can be arbitrarily substituted with glycosides.

[0415] In some embodiments, MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP are optionally replaced with glycosides, respectively.

[0416] In some embodiments, the glycoside is selected from glucuronides, galactosides, glucosides, mannosides, fucosides, O-GlcNAc, and O-GalNAc.

[0417] In a particular embodiment, T 1 , T 2 , T 3 , T 4 , T 5 and T 6 and V 1 , V 2 , V 3 , V 4 , V 5 , and V 6 The following can be selected: During the ceremony, T 1 (C1-C 12 ) is alkyl, V 1 is -CO-; T 2 (AA) p V 2 is non-existent (e.g., covalent bond); T 3 PABC and V 3 is non-existent (e.g., covalent bond); p is an integer between 1 and 10; d, e, and f are each 0; or During the ceremony, T 1 (C1-C 12 ) is alkyl, V 1 is -CONH-; T 2 (PEG) n V 2 is -CO-; T 3 (AA) p V 3 is non-existent (e.g., covalent bond); T 4 PABC and V 4 is non-existent (e.g., covalent bond); p is an integer between 1 and 10; e and f are 0, respectively; or During the ceremony, T 1 (C1-C 12 ) is alkyl, V 1 is -CO-; T 2 It is an amino acid analog, V 2 is -NH-; T 3 (PEG) n V 3 is -CO-; T 4 (AA) p V 4 is non-existent (e.g., covalent bond); T 5 PABC and V 5 is non-existent (e.g., covalent bond); p is an integer between 1 and 10; f is 0; or During the ceremony, T 1 (C1-C 12 ) is alkyl, V 1 is -CONH-; T 2 (PEG) n V 2 is -CO-; T 3 (AA) p V 3 is non-existent (e.g., covalent bond); T 4 PABC and V 4 is non-existent (e.g., covalent bond); p is an integer between 1 and 10; e and f are 0, respectively; or During the ceremony, T 1 (C1-C 12 ) is alkyl, V 1 is -CONH-; T 2 is a substitution (C1-C 12 ) is alkyl, V2 is -CO-; T 3 (AA) p V 3 is non-existent (e.g., covalent bond); T 4 PABC and V 4 is non-existent (e.g., covalent bond); p is an integer between 1 and 10; e and f are 0, respectively; or During the ceremony, T 1 (C1-C 12 ) is alkyl, V 1 is -CONH-; T 2 (PEG) n V 2 is -CO-; T 3 (AA) p V 3 is non-existent (e.g., covalent bond); T 4 It is PABA, and V 4 is -CO-; T 5 (C1-C 12 ) is alkyl, V 5 is non-existent (e.g., covalent bond); p is an integer between 1 and 10; f is 0; or During the ceremony, T 1 (C1-C 12 ) is alkyl, V 1 is -CO-; T 2 It is 4AP, V 2 is -CO-; T 3 (C1-C 12 ) is alkyl, V 3 is -CO-; T 4 (AA) p V 4is non-existent (e.g., covalent bond); T 5 PABC and V 5 is non-existent (e.g., covalent bond); p is an integer between 1 and 10; f is 0; or During the ceremony, T 1 (C1-C 12 ) is alkyl, V 1 is -CO-; T 2 It is 4AP, V 2 is -CO-; T 3 (C1-C 12 ) is alkyl, V 3 is -O-; T 4 (C1-C 12 ) is alkyl, V 4 is -CO-; T 5 (AA) p V 5 is non-existent (e.g., covalent bond); p is an integer between 1 and 10; T 6 PABC and V 6 is non-existent (e.g., covalent bond); or During the ceremony, T 1 (C1-C 12 ) is alkyl, V 1 is -CO-; T 2 It is an amino acid analog, V 2 is non-existent (e.g., covalent bond); T 3 (AA) p V 3 is non-existent (e.g., covalent bond); T 4 PABC and V 4 is non-existent (e.g., covalent bond); p is an integer between 1 and 10; e and f are 0, respectively; or During the ceremony, T 1 (C1-C 12 ) is alkyl, V 1 is -CONH-; T 2 (PEG) n V 2 is -CONH-; T 3 is a substitution (C1-C 12 ) is alkyl, V 3 is -CO-; T 4 (AA) p V 4 is non-existent (e.g., covalent bond); T 5 PABC and V 5 is non-existent (e.g., covalent bond); p is an integer between 1 and 10; f is 0; or During the ceremony, T 1 (C1-C 12 ) is alkyl, V 1 is -CO-; T 2 (AA) p V 2 is -NH-; T 3 (PEG) n V 3 is -CO-; T 4 (AA) p V 4 is non-existent (e.g., covalent bond); T 5 PABC and V 5 is non-existent (e.g., covalent bond); p is an integer between 1 and 10; f is 0; or During the ceremony, T 1 (C1-C 12 ) is alkyl, V1 is -CONH-; T 2 (PEG) n V 2 is -CO-; T 3 (AA) p V 3 is non-existent (e.g., covalent bond); T 4 It is PAP, and V 4 is -C(O)O-; p is an integer between 1 and 10; e and f are 0, respectively; or During the ceremony, T 1 (C1-C 12 ) is alkyl, V 1 is -CONH-; T 2 is a substitution (C1-C 12 ) is alkyl, V 2 is -CO-; T 3 (AA) p V 3 is non-existent (e.g., covalent bond); T 4 PABC and V 4 is non-existent (e.g., covalent bond); p is an integer between 1 and 10; e and f are 0, respectively; or During the ceremony, T 1 (C1-C 12 ) is alkyl, V 1 is -CONH-; T 2 is a substitution (C1-C 12 ) is alkyl, V 2 is -CO-; T 3 PABC and V 3 is non-existent (e.g., covalent bond); (d, e, and f are all 0).

[0418] In a particular embodiment, the first linker L A The left side of the above linker structure is bonded to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation portion, forming the first linker L A The right side of the above linker structure is bound to the first drug or active agent.

[0419] As described above, in some embodiments, L B is, -(T 7 -V 7 ) g -(T 8 -V 8 ) h -(T 9 -V 9 ) i -(T 10 -V 10 ) j -(T 11 -V 11 ) k -(T 12 -V 12 ) l -(T 13 -V 13 ) m -A second linker containing (wherein h, i, j, k, l, and m are independently 0 or 1, except that at least one of g, h, i, j, k, l, and m is 1).

[0420] In some embodiments, the second linker L B In: T 7 (C1-C 12 )alkyl and substituted (C1-C 12 ) Selected from alkyl groups; T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 (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 OH) x -, independently selected from 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, acetal group, disulfide, hydrazine, and ester; V 7 , V 8 , V 9 , V 10 , V 11 , V 12 , and V 13 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-, -SO2NR 15 -, -NR 15 The molecules are independently selected from SO2- and -P(O)OH-, where q is an integer between 1 and 6; (PEG) n teeth, [ka] And n is an integer from 1 to 30; The EDA has the following structure: [ka] The ethylenediamine moiety has , where y is an integer from 1 to 6, and r is 0 or 1; 4-amino-piperidine (4AP) is, [ka] and; AA is an amino acid residue, and p is an integer between 1 and 20; Each R 12 R is independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, and any two adjacent R 12 The groups can be linked in a cyclic manner to form a piperazinyl ring; Each R 13 This is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl; Each R 15 This is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

[0421] Any convenient tether group, T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 It can be used for the following purposes. For example, T 1 , T 2 , T 3 , T 4 , T 5 and T 6 Regarding the above, any of the tether groups, tether group T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 It can be used for that purpose.

[0422] Any convenient tether group, V 7 , V 8 , V 9 , V 10 , V 11 , V 12 , and V 13 It can be used for V. 1 , V2 , V 3 , V 4 , V 5 , and V 6 Regarding the above-mentioned linked functional groups, linked functional group V 7 , V 8 , V 9 , V 10 , V 11 , V 12 , and V 13 It can be used for this purpose.

[0423] In a specific method of operation, each R 13 R is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. In these embodiments, alkyl, substituted alkyl, aryl, and substituted aryl are R 13 The above is true.

[0424] In a specific method of operation, each R 15 R is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, 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, heterocyclyl, and substituted heterocyclyl is R 15 The above applies to these embodiments. In these embodiments, various possible substituents are R 15 The above is true.

[0425] Second Linker L B In a particular embodiment, the tether group T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13One or more of these are optionally substituted with glycosides or glycoside derivatives. In certain embodiments, the glycoside or glycoside derivative is selected from glucuronides, galactosides, glucosides, mannosides, fucosides, O-GlcNAc, and O-GalNAc.

[0426] Second Linker L B In certain embodiments, the MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP tether structures shown above may be substituted with one or more additional groups selected from glycosides and glycoside derivatives. For example, in some embodiments of the MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP tether structures shown above, the phenyl ring may be substituted with one or more additional groups selected from glycosides and glycoside derivatives. In certain embodiments, the glycoside or glycoside derivative may be selected from glucuronides, galactosides, glucosides, mannosides, fucosides, O-GlcNAc, and O-GalNAc.

[0427] In some embodiments, T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 These can be arbitrarily substituted with glycosides.

[0428] In some embodiments, MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP are optionally replaced with glycosides, respectively.

[0429] In some embodiments, the glycoside is selected from glucuronides, galactosides, glucosides, mannosides, fucosides, O-GlcNAc, and O-GalNAc.

[0430] L B In some embodiments, g, h, i, j, and k are each 1; l and m are both 0; T 7 It is a covalent bond; T 8 , T 9 , T 10 , T 11 and T 12 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 OH) x -, independently selected from 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, acetal group, hydrazine, and ester; V 7 , V 8 , V 9 , V 10 , V 11 and V 12 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-, -SO2NR 15 -, -NR 15 Independently selected from the group consisting of SO2- and -P(O)OH-; (PEG) n teeth, [ka] And n is an integer from 1 to 30; The EDA has the following structure: [ka] The ethylenediamine moiety has , where y is an integer from 1 to 6, and r is 0 or 1; 4-amino-piperidine (4AP) is, [ka] and; Each R 12 R is independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, and any two adjacent R 12 The groups can be linked in a cyclic manner to form a piperazinyl ring.

[0431] In some embodiments, T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , and T 12 These can be arbitrarily substituted with glycosides.

[0432] In some embodiments, MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP are optionally replaced with glycosides, respectively.

[0433] In some embodiments, the glycoside is selected from glucuronides, galactosides, glucosides, mannosides, fucosides, O-GlcNAc, and O-GalNAc.

[0434] In a particular embodiment, T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 and V 7 , V 8 , V 9 , V 10 , V 11 , V 12, and V 13 The following can be selected: During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CO-; T 9 (AA) p V 9 is non-existent (e.g., covalent bond); T 10 PABC and V 10 is non-existent (e.g., covalent bond); k, l, and m are each 0; or During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CONH-; T 9 (PEG) n V 9 is -CO-; T 10 (AA) p V 10 is non-existent (e.g., covalent bond); T 11 PABC and V 11 is non-existent (e.g., covalent bond); l and m are 0, respectively; or During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CO-; T9 It is an amino acid analog, V 9 is -NH-; T 10 (PEG) n V 10 is -CO-; T 11 (AA) p V 11 is non-existent (e.g., covalent bond); T 12 PABC and V 12 is non-existent (e.g., covalent bond); m is 0; or During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CONH-; T 9 (PEG) n V 9 is -CO-; T 10 (AA) p V 10 is non-existent (e.g., covalent bond); T 11 PABC and V 11 is non-existent (e.g., covalent bond); l and m are 0, respectively; or During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CONH-; T 9 is a substitution (C1-C 12 ) is alkyl, V 9 is -CO-; T10 (AA) p V 10 is non-existent (e.g., covalent bond); T 11 PABC and V 11 is non-existent (e.g., covalent bond); l and m are 0, respectively; or During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CONH-; T 9 (PEG) n V 9 is -CO-; T 10 (AA) p V 10 is non-existent (e.g., covalent bond); T 11 It is PABA, and V 11 is -CO-; T 12 (C1-C 12 ) is alkyl, V 12 is non-existent (e.g., covalent bond); m is 0; or During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CO-; T 9 It is 4AP, V 9 is -CO-; T 10 (C1-C 12 ) is alkyl, V 10 is -CO-; T11 (AA) p V 11 is non-existent (e.g., covalent bond); T 12 PABC and V 12 is non-existent (e.g., covalent bond); m is 0; or During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CO-; T 9 It is 4AP, V 9 is -CO-; T 10 (C1-C 12 ) is alkyl, V 10 is -O-; T 11 (C1-C 12 ) is alkyl, V 11 is -CO-; T 12 (AA) p V 12 is non-existent (e.g., covalent bond); T 13 PABC and V 13 is non-existent (e.g., covalent bond); or During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CO-; T 9 It is an amino acid analog, V 9 is non-existent (e.g., covalent bond); T 10 (AA) p V10 is non-existent (e.g., covalent bond); T 11 PABC and V 11 is non-existent (e.g., covalent bond); l and m are 0, respectively; or During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CONH-; T 9 (PEG) n V 9 is -CONH-; T 10 is a substitution (C1-C 12 ) is alkyl, V 10 is -CO-; T 11 (AA) p V 11 is non-existent (e.g., covalent bond); T 12 PABC and V 12 is non-existent (e.g., covalent bond); m is 0; or During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CO-; T 9 (AA) p V 9 is -NH-; T 10 (PEG) n V 10 is -CO-; T 11 (AA) pV 11 is non-existent (e.g., covalent bond); T 12 PABC and V 12 is non-existent (e.g., covalent bond); m is 0; or During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CONH-; T 9 (PEG) n V 9 is -CO-; T 10 (AA) p V 10 is non-existent (e.g., covalent bond); T 11 It is PAP, and V 11 is -C(O)O-; l and m are 0, respectively; or During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CO-; T 9 (AA) p V 9 is non-existent (e.g., covalent bond); T 10 PABC and V 10 is non-existent (e.g., covalent bond); T 11 It is PAP, and V 11 is -C(O)O-; l and m are 0, respectively; or During the ceremony, T7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is -CONH-; T 9 is a substitution (C1-C 12 ) is alkyl, V 9 is -CO-; T 10 PABC and V 10 is non-existent (e.g., covalent bond); k, l, and m are each 0; or During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is non-existent (e.g., covalent bond); T 9 It is a heteroaryl compound, and V 9 is non-existent (e.g., covalent bond); T 10 (C1-C 12 ) is alkyl, V 10 is -CONH-; T 11 (PEG) n V 11 is -CO-; l and m are 0, respectively; or During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is non-existent (e.g., covalent bond); T 9 It is a heteroaryl compound, and V 9is non-existent (e.g., covalent bond); T 10 (C1-C 12 ) is alkyl, V 10 is -CONH-; T 11 is a substitution (C1-C 12 ) is alkyl, V 11 is -CO-; T 12 (AA) p V 12 is non-existent (e.g., covalent bond); T 13 PAB and V 13 is non-existent (e.g., covalent bond); or During the ceremony, T 7 is non-existent (e.g., covalent bond), V 7 It is -NHCO-; T 8 (C1-C 12 ) is alkyl, V 8 is non-existent (e.g., covalent bond); T 9 It is a heteroaryl compound, and V 9 is non-existent (e.g., covalent bond); T 10 (C1-C 12 ) is alkyl, V 10 is -CONH-; T 11 is a substitution (C1-C 12 ) is alkyl, V 11 is -CO-; T 12 (AA) p V 12 is non-existent (e.g., covalent bond); T 13 PABC and V 13 It is non-existent (for example, a covalent bond).

[0435] In certain embodiments, a second linker L BThe left side of the above linker structure is bonded to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation portion, and the second linker L B The right side of the above linker structure is bound to the first drug or active agent.

[0436] In certain embodiments, the conjugate is an antibody-drug conjugate in which a ROR antibody, e.g., an activatable ROR antibody and a drug are bound by a linker, as described above. In some cases, the linker (e.g., L) A and / or L B A linker is a cleavable linker. A cleavable linker is a linker that contains one or more cleavable moieties, each containing one or more bonds that can dissociate under certain conditions, thereby separating the cleavable linker into two or more separable moieties. For example, each cleavable moiety may contain one or more covalent bonds that, under certain conditions, can dissociate or decompose to separate the cleavable linker into two or more moieties. Thus, a linker contained in an antibody-drug conjugate can be a cleavable linker such that, under suitable conditions, the cleavable linker is cleaved, allowing the drug to be separated or released from the antibody at a desired target site of action.

[0437] In some cases, a cleavable linker includes two cleavable segments (e.g., a first cleavable segment and a second cleavable segment). The cleavable segments can be configured such that cleavage of both segments is required to separate or release the drug from a ROR antibody, e.g., an activatable ROR antibody, at a desired target site of the drug's action. For example, cleavage of a cleavable linker can be achieved by first cleaving one of the two cleavable segments and then cleaving the other of the two cleavable segments. In certain embodiments, a cleavable linker includes a first cleavable segment and a second cleavable segment that prevents cleavage of the first cleavable segment. "Prevents cleavage" means that the presence of an uncleaved second cleavable segment reduces or substantially inhibits the possibility of cleavage of the first cleavable segment, thereby substantially reducing or preventing the amount of cleavable linker. For example, the presence of an uncleaved second cleavable segment may prevent cleavage of the first cleavable segment. The presence of a second cleavable region prevents the cleavage of the first cleavable region, which then significantly reduces the amount of drug released from the antibody or prevents its release altogether. For example, early release of the drug from the antibody can be significantly reduced or blocked until the antibody-drug conjugate reaches or near the desired target site of the drug's action.

[0438] In some cases, the second cleavable portion prevents the cleavage of the first cleavable portion, so cleavage of the cleavable linker can be achieved by first cleaving the second cleavable portion and then the first cleavable portion. Cleavage of the second cleavable portion reduces or eliminates the interference with cleavage of the first cleavable portion, thereby enabling cleavage of the first cleavable portion. Cleavage of the first cleavable portion allows the cleavable linker to dissociate or separate into two or more parts, as described above, and release of the drug from the antibody-drug conjugate. In some cases, cleavage of the first cleavable portion does not substantially occur in the presence of the uncleaved second cleavable portion. In effect, this means that approximately 10% or less of the cutting of the first cleavable portion occurs in the presence of the uncut second cleavable portion, and for example, approximately 9% or less, or approximately 8% or less, or approximately 7% or less, or approximately 6% or less, or approximately 5% or less, or approximately 4% or less, or approximately 3% or less, or approximately 2% or less, or approximately 1% or less, or approximately 0.5% or less, or approximately 0.1% or less of the cutting of the first cleavable portion occurs in the presence of the uncut second cleavable portion.

[0439] In other words, the second cleavable portion may protect the first cleavable portion from cleavage. For example, the presence of an uncleaved second cleavable portion protects the first cleavable portion from cleavage, thereby significantly reducing or preventing premature release of the drug from the antibody until the antibody-drug conjugate reaches or near the desired target site of drug action. Thus, cleavage of the second cleavable portion exposes the first cleavable portion (e.g., releases its protection), making it possible to cleave the first cleavable portion, resulting in the cleavage of the cleavable linker, and then, as described above, the drug is separated or released from the antibody at the desired target site of drug action. In certain cases, cleavage of the second cleavable portion exposes the first cleavable portion to subsequent cleavage, but cleavage of the second cleavable portion does not, by itself, result in the cleavage of the cleavable linker (e.g., cleavage of the first cleavable portion is still required to cleave the cleavable linker).

[0440] Each cleavable portion within a cleavable linker may be an enzymatically cleavable portion. For example, the first cleavable portion may be a first enzymatically cleavable portion, and the second cleavable portion may be a second enzymatically cleavable portion. An enzymatically cleavable portion is a cleavable portion that can be separated into two or more of the above-mentioned portions by the enzymatic action of an enzyme. An enzymatically cleavable portion may be any portion that can be cleaved by the enzymatic action of an enzyme (e.g., esters, peptides, glycosides, etc., but not limited to these). In some cases, the enzyme that cleaves the enzymatically cleavable portion is present at the desired target site of action (e.g., the desired target site of the drug to be released from the antibody-drug conjugate). In some cases, the enzyme that cleaves the enzymatically cleavable portion is not present in significant amounts in other regions (e.g., whole blood, plasma, serum). Therefore, when the enzymatically cleavable portion is cleaved, substantial cleavage occurs at the desired site of action, but it can be controlled so that significant cleavage does not occur in other regions or before the antibody-drug conjugate reaches the desired site of action.

[0441] For example, as described herein, the antibody-drug conjugates of this disclosure can be used for the treatment of cancer (e.g., to deliver cancer therapeutics to desired sites of action where cancer cells are present). In some cases, enzymes (e.g., esterases that cleave ester bonds and glycosidases that cleave glycosidic bonds) may be cancer biomarkers that are overexpressed in cancer cells. Overexpression of specific enzymes in cancer, and their localization therein, can be used in conjunction with an enzymatically cleavable moiety contained in the cleavable linker of the antibody-drug conjugate of this disclosure to specifically release a drug at a desired site of action (e.g., the site of cancer (and the overexpressed enzyme)). Thus, in some embodiments, the enzymatically cleavable moiety is a cleavable moiety (e.g., an ester or glycoside) that can be cleaved by an enzyme overexpressed in cancer cells. For example, the enzyme may be an est...

Claims

1. An antibody-drug conjugate (ADC) of formula (I), a. Activatable antibodies that bind to tyrosine-protein kinase membrane receptors (RORs); and b. Two or more drugs conjugated to the pyridazine-pyrrolo coupling portion via a linker. The antibody-drug conjugate, including 【Chemical 85】 (In the formula, Ab represents an antibody that binds to ROR, optionally an activatable antibody that binds to ROR, and optionally the activatable antibody comprises (a) a masking peptide comprising a masking moiety (MM) and a cleavable moiety (CM); and (b) a target binding site (TBM); Z １ Z ２ , and Z ４ Each of them operates independently, CR ４ And; Z ３ C-L Ｂ -W ２ And; R １ 、R ２ 、R ３ 、and R ４ are each selected from hydrogen and alkyl; L Ａ teeth, -(T １ -V １ ) ａ -(T ２ -V ２ ) ｂ -(T ３ -V ３ ) ｃ -(T ４ -V ４ ) ｄ -(T ５ -V ５ ) ｅ -(T ６ -V ６ ) ｆ - The first linker includes, a, b, c, d, e, and f are each independently either 0 or 1, provided that at least one of a, b, c, d, e, and f is 1; T １ , T ２ , T ３ , T ４ , T ５ and T ６ 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 OH) ｘ -, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxyphenyl (PHP), acetal group, hydrazine, disulfide, and ester, each independently selected from -, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), para-amino-benzyloxycarbonyl (PABC), para-amino-benzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxyphenyl (PHP), acetal group, hydrazine, disulfide, and ester, where EDA is the ethylenediamine moiety, PEG is polyethylene glycol, AA is an amino acid residue or amino acid analog, each w is an integer from 1 to 20, each n is an integer from 1 to 30, each p is an integer from 1 to 20, and each x is an integer from 1 to 12; V １ , V ２ , V ３ , V ４ , V ５ , and V ６ is a covalent bond, -CO-, -NR １５ -, -NR １５ (CH ２ ) ｑ -, -NR １５ (C ６ H ４ )-,-CONR １５ -, -NR １５ CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO ２ -, -SO ２ NR １５ -, -NR １５ SO ２ Each of the groups consisting of - and -P(O)OH- is independently selected, where each q is an integer from 1 to 6; Each R １３ is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl; Each R １５ This is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; L Ｂ teeth, -(T ７ -V ７ ) ｇ -(T ８ -V ８ ) ｈ -(T ９ -V ９ ) ｉ -(T １０ -V １０ ) ｊ -(T １１ -V １１ ) ｋ -(T １２ -V １２ ) ｌ -(T １３ -V １３ ) ｍ - It is a second linker that includes, g, h, i, j, k, l, and m are each independently 0 or 1, provided that at least one of g, h, i, j, k, l, and m is 1; T ７ 、 T ８ 、 T ９ 、 T １０ 、 T １１ 、 T １２ 、 and T １３ are each independently selected from a covalent bond, (C １ -C １２ ), alkyl, substituted (C １ -C １２ ), aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA) ｗ 、(PEG) ｎ 、(AA) ｐ 、-(CR １３ OH) ｘ -、4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxy-phenyl (PHP), an acetal group, hydrazine, disulfide, and an ester, where EDA is an ethylenediamine moiety, PEG is polyethylene glycol, AA is an amino acid residue or an amino acid analog, each w is an integer from 1 to 20, each n is an integer from 1 to 30, each p is an integer from 1 to 20, and each x is an integer from 1 to 12; V ７ , V ８ , V ９ , V １０ , V １１ , V １２ , and V １３ is a covalent bond, -CO-, -NR １５ -, -NR １５ (CH ２ ) ｑ -, -NR １５ (C ６ H ４ )-,-CONR １５ -, -NR １５ CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO ２ -, -SO ２ NR １５ -, -NR １５ SO ２ Each of the groups consisting of - and -P(O)OH- is independently selected, where each q is an integer from 1 to 6; Each R １３ is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl; Each R １５ This is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; s is an integer from 1 to 10, and is arbitrarily 4; W １ It is the first drug; W ２ (This is the second drug.)

2. T １ is, (C １ -C １２ ) alkyl and substituted (C １ -C １２ ) Selected from alkyl; T ２ , T ３ , T ４ , T ５ and T ６ 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 OH) ｘ -, 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, acetal group, hydrazine, and ester, each independently selected; V １ , V ２ , V ３ , V ４ , V ５ , and V ６ is a covalent bond, -CO-, -NR １５ -, -NR １５ (CH ２ ) ｑ -, -NR １５ (C ６ H ４ )-,-CONR １５ -, -NR １５ CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO ２ -, -SO ２ NR １５ -, -NR １５ SO ２ They are independently selected from the group consisting of - and -P(O)OH-; (PEG) ｎ teeth, 【Chemical 86】 And; EDA has the following structure: 【Chemistry 87】 It is an ethylenediamine moiety having; 4-amino-piperidine (4AP) is 【Chemical 88】 And; Each R １２ R is independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, and any two adjacent R １２ The groups can be linked in a cyclic manner to form a piperazinyl ring; q is an integer between 1 and 6; r is either 0 or 1; y is an integer between 1 and 6. The ADC according to claim 1.

3. T １ is, (C １ -C １２ ) is alkyl, V １ is -CONH-; T ２ is substitution (C １ -C １２ ) is alkyl, V ２ is -CO-; T ３ (AA) ｐ V ３ It is non-existent; T ４ PABC and V ４ It is non-existent; p is an integer between 1 and 10; a, b, c, and d are each 1; e and f are both 0. The ADC according to claim 1 or 2.

4. T ７ It is a covalent bond; T ８ , T ９ , T １０ , T １１ and T １２ 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 OH) ｘ -, 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, acetal group, hydrazine, and ester, each independently selected; V ７ , V ８ , V ９ , V １０ , V １１ and V １２ is a covalent bond, -CO-, -NR １５ -, -NR １５ (CH ２ ) ｑ -, -NR １５ (C ６ H ４ )-,-CONR １５ -, -NR １５ CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO ２ -, -SO ２ NR １５ -, -NR １５ SO ２ They are independently selected from the group consisting of - and -P(O)OH-; (PEG) ｎ teeth, 【Chemical 89】 And n is an integer from 1 to 30; EDA has the following structure: [Chemical 90] The ethylenediamine moiety has , where y is an integer from 1 to 6, and r is 0 or 1; 4-amino-piperidine (4AP) is 【Chemistry 91】 And; Each R １２ R is independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, and any two adjacent R １２ The groups can be linked in a cyclic manner to form a piperazinyl ring; Each of g, h, i, j, and k is 1; l and m are both 0. The ADC according to any one of claims 1 to 3.

5. T ７ V is non-existent, ７ It is -NHKO-; T ８ is, (C １ -C １２ ) is alkyl, V ８ is -CONH-; T ９ is substitution (C １ -C １２ ) is alkyl, V ９ is -CO-; T １０ (AA) ｐ V １０ It is non-existent; T １１ PABC and V １１ It is non-existent; p is an integer between 1 and 10; Each of g, h, i, j, and k is 1; l and m are both 0. The ADC according to any one of claims 1 to 4.

6. T ２ and T ９ One or both of these are -NHCO (PEG) ｔ Replaced with (C １ -C ６ ) is alkylene, (PEG) ｔ teeth, 【Chemistry 92】 The ADC according to any one of claims 1 to 5, wherein t is an integer from 2 to 10, and is arbitrarily 8.

7. T ３ and T １０ The ADC according to any one of claims 1 to 6, wherein one or both of the elements have 2 p.

8. The ADC according to any one of claims 1 to 7, wherein s is 4.

9. The ADC according to any one of claims 1 to 8, wherein MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP are each optionally substituted with a glycoside, and optionally the glycoside is selected from glucuronide, galactoside, glucoside, mannoside, fucoside, O-GlcNAc, and O-GalNAc.

10. W １ and W ２ The ADC according to any one of claims 1 to 9, wherein one or both of are camptothecin analogs, and optionally the camptothecin analog is berotecan.

11. W １ and W ２ Each of them is belotecan, according to any one of claims 1 to 10.

12. Formula (II): 【Chemistry 93】 (In the formula, Ab represents an antibody that binds to ROR, optionally an activatable antibody that binds to ROR, and optionally the activatable antibody comprises (a) a masking peptide comprising a masking moiety (MM) and a cleavable moiety (CM); and (b) a target binding site (TBM); (s is an integer from 1 to 10, and is arbitrarily 4.) ADC represented by

13. The ADC according to any one of claims 1 to 12, wherein the TBM of the activatable antibody comprises an antibody light chain variable (VL) region and an antibody heavy chain variable (VH) region, the VH region comprising VH complementarity determination region 1 (CDR1), VH complementarity determination region 2 (CDR2), and VH complementarity determination region 3 (CDR3) as shown in VH comprising the amino acid sequence of SEQ ID NO: 25, and the VL region comprising VL CDR1, VL CDR2, and VL CDR3 as shown in VL comprising the amino acid sequence of SEQ ID NO: 26; and the activatable antibody is capable of binding to ROR when the CM is cleaved.

14. Ab is, Masking peptide, antibody light chain variable (VL) region, and antibody heavy chain variable (VH) region The masking peptide comprises any one amino acid sequence of SEQ ID NOs: 29, 30, or 31. The VH region includes VH complementarity determination region 1 (CDR1), VH complementarity determination region 2 (CDR2), and VH complementarity determination region 3 (CDR3) as shown in VH containing the amino acid sequence of SEQ ID NO: 25, and the VL region includes VL CDR1, VL CDR2, and VL CDR3 as shown in VL containing the amino acid sequence of SEQ ID NO: 26; The activatable antibody or fragment thereof optionally comprises a polypeptide from the N-terminus to the C-terminus that includes the masking peptide and the antibody VL region. The ADC according to any one of claims 1 to 13.

15. Ab is, (1) VH CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 7, 12, 13, 18, and 27; (2) VH CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 8, 14, 19, and 24; and (3) VH CDR3 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 9, 15, 20, and 28 The VH region including; and (1) VL CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 10, 16, and 21; (2) VL CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 11, and 22; and (3) VL CDR3 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 17, and 23 VL area including An ADC according to any one of claims 1 to 14, including the ADC described in any one of claims 1 to 14.

16. Ab is, (i) VH regions including VH CDR1 containing the amino acid sequence of SEQ ID NO: 1, VH CDR2 containing the amino acid sequence of SEQ ID NO: 2, and VH CDR3 containing the amino acid sequence of SEQ ID NO: 3; and VL regions including VL CDR1 containing the amino acid sequence of SEQ ID NO: 4, VL CDR2 containing the amino acid sequence of SEQ ID NO: 5, and VL CDR3 containing the amino acid sequence of SEQ ID NO: 6; (ii) A VH region comprising VH CDR1 containing the amino acid sequence of SEQ ID NO: 7, VH CDR2 containing the amino acid sequence of SEQ ID NO: 8, and VH CDR3 containing the amino acid sequence of SEQ ID NO: 9; and a VL region comprising VL CDR1 containing the amino acid sequence of SEQ ID NO: 10, VL CDR2 containing the amino acid sequence of SEQ ID NO: 11, and VL CDR3 containing the amino acid sequence of SEQ ID NO: 6; (iii) A VH region containing VH CDR1 containing the amino acid sequence of SEQ ID NO: 12, VH CDR2 containing the amino acid sequence of SEQ ID NO: 2, and VH CDR3 containing the amino acid sequence of SEQ ID NO: 3; and a VL region containing VL CDR1 containing the amino acid sequence of SEQ ID NO: 4, VL CDR2 containing the amino acid sequence of SEQ ID NO: 5, and VL CDR3 containing the amino acid sequence of SEQ ID NO: 6; (iv) A VH region comprising VH CDR1 containing the amino acid sequence of SEQ ID NO: 13, VH CDR2 containing the amino acid sequence of SEQ ID NO: 14, and VH CDR3 containing the amino acid sequence of SEQ ID NO: 15; and a VL region comprising VL CDR1 containing the amino acid sequence of SEQ ID NO: 16, VL CDR2 containing the amino acid sequence of SEQ ID NO: 11, and VL CDR3 containing the amino acid sequence of SEQ ID NO: 17; (v) A VH region comprising VH CDR1 containing the amino acid sequence of SEQ ID NO: 18, VH CDR2 containing the amino acid sequence of SEQ ID NO: 19, and VH CDR3 containing the amino acid sequence of SEQ ID NO: 20; and a VL region comprising VL CDR1 containing the amino acid sequence of SEQ ID NO: 21, VL CDR2 containing the amino acid sequence of SEQ ID NO: 22, and VL CDR3 containing the amino acid sequence of SEQ ID NO: 23; (vi) A VH region including VH CDR1 containing the amino acid sequence of SEQ ID NO: 1, VH CDR2 containing the amino acid sequence of SEQ ID NO: 24, and VH CDR3 containing the amino acid sequence of SEQ ID NO: 3; and a VL region including VL CDR1 containing the amino acid sequence of SEQ ID NO: 4, VL CDR2 containing the amino acid sequence of SEQ ID NO: 5, and VL CDR3 containing the amino acid sequence of SEQ ID NO: 6; or (vii) VH region including VH CDR1 containing the amino acid sequence of SEQ ID NO: 27, VH CDR2 containing the amino acid sequence of SEQ ID NO: 24, and VH CDR3 containing the amino acid sequence of SEQ ID NO: 28; and VL region including VL CDR1 containing the amino acid sequence of SEQ ID NO: 4, VL CDR2 containing the amino acid sequence of SEQ ID NO: 5, and VL CDR3 containing the amino acid sequence of SEQ ID NO: 17 An ADC according to any one of claims 1 to 15, including the ADC described in any one of claims 1 to 15.

17. The ADC according to any one of claims 1 to 16, wherein Ab further comprises the framework 1 (FR1), framework 2 (FR2), framework 3 (FR3), and / or framework 4 (FR4) sequences.

18. The ADC according to any one of claims 1 to 17, wherein Ab further comprises a human framework sequence, optionally FR1, FR), FR3 and / or FR) sequence as shown in SEQ ID NO: 25 or 26.

19. The ADC according to any one of claims 14 to 18, wherein VH comprises the amino acid sequence of SEQ ID NO: 25, and VL comprises the amino acid sequence of SEQ ID NO:

26.

20. The ADC according to any one of claims 14 to 19, wherein the masking peptide comprises the amino acid sequence of SEQ ID NO:

29.

21. The ADC according to claim 20, wherein Ab comprises the polypeptide, and the polypeptide comprises the amino acid sequence of SEQ ID NO: 34 or 35.

22. The ADC according to any one of claims 14 to 19, wherein the masking peptide comprises the amino acid sequence of SEQ ID NO:

30.

23. The ADC according to claim 22, wherein Ab comprises the polypeptide, and the polypeptide comprises the amino acid sequence of SEQ ID NO: 36 or 37.

24. The ADC according to any one of claims 14 to 19, wherein the masking peptide comprises the amino acid sequence of SEQ ID NO:

31.

25. The ADC according to claim 24, wherein Ab comprises the polypeptide, and the polypeptide comprises the amino acid sequence of SEQ ID NO: 38 or 39.

26. The ADC according to any one of claims 1 to 25, wherein Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 32 or a variant thereof that has been manipulated to be conjugated into a linker payload, or SEQ ID NO: 42 or a variant thereof that has been conjugated into the linker payload.

27. The antibody Ab has the sequence of formula (VIII) X １ (fGly’)X ２ Z ２０ X ３ Z ３０ (VIII) (In the formula, fGly' is an amino acid residue coupled to the drug via a linker; Z ２０ is either a proline (P) or alanine (A) residue; Z ３０ is a basic amino acid residue optionally selected from the group consisting of arginine (R), lysine (K), and histidine (H), or an aliphatic amino acid residue optionally selected from the group consisting of alanine (A), glycine (G), leucine (L), valine (V), isoleucine (I), and proline (P); X １ X may or may not be present, and if present, it may be any amino acid residue, provided that the sequence of formula (VIII) is at the N-terminus of antibody Ab. １ It exists; X ２ and X ３ These can be any amino acid residue, independently. Optionally, The sequence of formula (VIII) is L(fGly')TPSR (sequence number 146), M(fGly')TPSR (sequence number 147), V(fGly')TPSR (sequence number 148), L(fGly')SPSR (sequence number 149), L(fGly')APSR (sequence number 150), L(fGly')VPSR (sequence number 151), L(fGly')GPSR (sequence number 152), I(fGly')TPAR (sequence number 153), L(fGly')TPSK (sequence number 154), M(fGly')TPSK (sequence number 155), V(fGly')TPSK (sequence number 156), L(fGly')SP Selected from the group consisting of SK (SEQ ID NO: 157), L(fGly')APSK (SEQ ID NO: 158), L(fGly')VPSK (SEQ ID NO: 159), L(fGly')GPSK (SEQ ID NO: 160), L(fGly')TPSA (SEQ ID NO: 161), I(fGly')TPAA (SEQ ID NO: 162), M(fGly')TPSA (SEQ ID NO: 163), V(fGly')TPSA (SEQ ID NO: 164), L(fGly')SPSA (SEQ ID NO: 165), L(fGly')APSA (SEQ ID NO: 166), L(fGly')VPSA (SEQ ID NO: 167), and L(fGly')GPSA (SEQ ID NO: 168); Furthermore, optionally, the sequence of formula (VIII) includes L(fGly')TPSR (sequence number 146), The ADC according to any one of claims 1 to 26.

28. The ADC according to any one of claims 1 to 27, wherein Ab is an IgG1 antibody, or optionally an IgG1 kappa antibody.

29. The ADC according to any one of claims 1 to 28, wherein Ab comprises a heavy chain comprising one or more amino acid sequences of sequence number 32 or 169 or a variant thereof, and a light chain comprising one of the amino acid sequences of sequence number 34, 36, or 38.

30. The ADC according to any one of claims 1 to 29, wherein Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO:

48.

31. The ADC according to any one of claims 1 to 29, wherein Ab comprises a heavy chain containing the amino acid sequence of SEQ ID NO:

171.

32. The ADC according to any one of claims 1 to 31, wherein Ab comprises a light chain having one of the amino acid sequences of SEQ ID NOs: 34, 36, or 38.

33. Ab is as follows (i) to (vi): (i) A heavy chain containing the amino acid sequence shown in SEQ ID NO: 48 and a light chain containing the amino acid sequence shown in SEQ ID NO: 34; (ii) A heavy chain containing the amino acid sequence shown in SEQ ID NO: 48 and a light chain containing the amino acid sequence shown in SEQ ID NO: 36; (iii) A heavy chain containing the amino acid sequence shown in SEQ ID NO: 48 and a light chain containing the amino acid sequence shown in SEQ ID NO: 38; (iv) A heavy chain containing the amino acid sequence shown in SEQ ID NO: 171 and a light chain containing the amino acid sequence shown in SEQ ID NO: 34; (v) A heavy chain containing the amino acid sequence shown in SEQ ID NO: 171 and a light chain containing the amino acid sequence shown in SEQ ID NO: 36; (vi) A heavy chain containing the amino acid sequence shown in SEQ ID NO: 171 and a light chain containing the amino acid sequence shown in SEQ ID NO: 38 An ADC according to any one of claims 1 to 32, comprising any one of the above.

34. The ADC according to any one of claims 1 to 33, wherein Ab is a monoclonal antibody.

35. The ADC according to any one of claims 1 to 34, wherein Ab is a humanized, human, or chimeric antibody.

36. Ab is Fab, Fab', F(ab') ２ , Fv, scFv, (scFv) ２ The ADC according to any one of claims 1 to 27, which is a multispecific antibody formed from a single-chain antibody molecule, a bivariable region antibody, a single variable region antibody, a linear antibody, a V region, or an antibody fragment.

37. The ADC according to any one of claims 1 to 35, wherein Ab is a multispecific antibody, and optionally the multispecific antibody is a bispecific antibody.

38. The ADC according to any one of claims 1 to 37, wherein Ab binds to both ROR1 and ROR2, optionally Ab binds to both human ROR1 and human ROR2, optionally Ab is a silent Fc (sFc), and optionally Ab comprises an sFc having the amino acid sequence shown in SEQ ID NO:

44.

39. A pharmaceutical composition comprising the ADC described in any one of claims 1 to 38 and a pharmaceutically acceptable excipient.

40. The pharmaceutical composition according to claim 39, characterized by an ADC drug-to-antibody ratio (DAR) of approximately 1 to approximately 20.

41. The pharmaceutical composition according to claim 40, wherein the DAR is approximately 2 to approximately 8.

42. The pharmaceutical composition according to claim 40 or 41, wherein the DAR is approximately 8.

43. An activated, activatable ADC produced by activating the ADC according to any one of claims 1 to 38.

44. An activated activatable ADC, which is an ADC according to any one of claims 1 to 38, wherein the activatable antibody Ab is activated.

45. The ADC or Ab is activated by treating the ADC containing the activatable antibody Ab with one or more proteases, wherein the Ab comprises a masking peptide, the one or more proteases cleave within the masking peptide, and optionally the one or more proteases comprise MMP-9, as per claim 43 or 44.

46. The ADC according to any one of claims 43 to 45, wherein Ab comprises a heavy chain having the amino acid sequence shown in SEQ ID NO: 48 or 171 and a light chain having the amino acid sequence shown in SEQ ID NO:

33.

47. Ab is, (i) a heavy chain having the amino acid sequence shown in SEQ ID NO: 48 and a light chain having the amino acid sequence shown in SEQ ID NO: 33; or (ii) A heavy chain containing the amino acid sequence shown in SEQ ID NO: 171 and a light chain containing the amino acid sequence shown in SEQ ID NO: 33 An ADC according to any one of claims 43 to 46, including the ADC described in any one of claims 43 to 46.

48. A method for treating a subject having cancer, the method comprising administering to the subject a therapeutically effective amount of an ADC according to any one of claims 1 to 38 and 43 to 47 or a pharmaceutical composition according to any one of claims 39 to 42.

49. The method according to claim 48, wherein the cancer is a cancer that expresses the ROR antigen.

50. The method according to claim 48 or 49, wherein the cancer expresses the ROR1 antigen.

51. The method according to claim 48 or 49, wherein the cancer expresses the ROR2 antigen.

52. The method according to claim 48 or 49, wherein the cancer expresses ROR1 antigen and ROR2 antigen.

53. The method according to any one of claims 48 to 52, wherein the cancer is selected from the group consisting of pancreatic cancer, ovarian cancer, breast cancer, lung cancer, gastric cancer, melanoma, Ewing's sarcoma, chronic lymphocytic leukemia, mantle cell lymphoma, B-ALL, hematological cancer, prostate cancer, colon cancer, kidney cancer, thyroid cancer, liver cancer, urothelial carcinoma, melanoma, endometrial cancer, clear cell renal cell carcinoma, clear cell carcinoma, and uterine cancer, and optionally the cancer is chronic myeloid leukemia, colorectal cancer, estrogen receptor-positive, progesterone receptor-positive and HER2-negative breast cancer, triple-negative breast cancer, or non-small cell lung cancer.

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