Antibody-drug conjugate of tissue factor and uses thereof

Abstract

The present invention provides anti-tissue factor antibody-drug conjugate structures. The antibody-drug conjugate structures include a branched linker, with two or more payloads attached to the antibody per branched linker. Additionally, the present disclosure encompasses compounds and methods for producing such conjugates. Additionally, the present disclosure encompasses methods of using the conjugates. In some embodiments, such TF-ADCs bind to the same epitope of human TF as antibodies comprising the heavy chain variable region (VH) and light chain variable region (VL) described herein.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 404,447, filed September 7, 2022, and U.S. Provisional Patent Application No. 63 / 498,233, filed April 25, 2023, the disclosures of each of which are incorporated herein by reference in their entirety.

[0002] Sequence Listing This application contains a computer-readable sequence listing, which has been submitted herewith in XML file format, the entire contents of which are incorporated herein by reference. The sequence listing XML file submitted herewith is named "14529-138-228_SEQ_LISTING.xml", was created on September 5, 2023, and is 191,857 bytes in size.

[0003] Field The present disclosure relates generally to antibody-drug conjugates (ADCs) that bind to tissue factor (TF, e.g., human TF) and methods of use thereof.

[0004] Introduction Blood coagulation involves a series of complex processes that result in blood clotting. Tissue factor (TF) plays a key role in these clotting processes. TF is the cell surface receptor for the serine protease factor VIIa (FVIIa). The TF / FVIIa complex catalyzes the conversion of inactive protease factor X (FX) to active protease factor Xa (FXa). FXa and its cofactor FVa form the prothrombinase complex, which generates thrombin from prothrombin. Thrombin converts soluble fibrinogen to insoluble fibrin chains and catalyzes many other coagulation-related processes. TF is overexpressed in several types of solid tumors. In cancer, TF / FVIIa signaling can support angiogenesis, tumor progression, and metastasis.

[0005] Antibody-drug conjugates (ADCs) have emerged as a new class of targeted delivery therapy over the past two decades. A typical ADC contains an antibody-based targeting moiety that is conjugated to a highly potent pharmaceutical (payload) via a chemical linker using available bioconjugation methods. The molar ratio of the targeting moiety (e.g., antibody) to the attached payload can be varied and is referred to as the drug-to-antibody ratio (DAR). Commonly used bioconjugation methods either utilize endogenous amino acid residues (i.e., lysine and cysteine) in proteins or rely on the selective engagement of bioorthogonal functional groups that have been intentionally introduced into proteins. As an example of the latter approach, the hydrazino-iso-Picteth-Spengler (HIPS) conjugation method (Figure 1) utilizes an aldehyde functional group ("aldehyde tag"), which can be introduced into proteins (e.g., antibodies) via various means (e.g., by the action of formyl-generating enzymes (FGEs)) and serves as a conjugation handle. The aldehyde group reacts completely with the HIPS indole moiety to form a stable carbon-carbon bond, which permanently attaches the payload of choice to the protein in a single chemical step.

[0006] There remains a need in the art for ADCs that can target TF to treat, prevent, or ameliorate TF-mediated diseases, disorders, or conditions, such as those involving tumor cells that express TF. Summary of the Invention

[0007] The present disclosure provides ADCs comprising antibodies that bind to tissue factor ("TF-ADCs"). In some embodiments, such TF-ADCs bind to the same epitope of human TF as antibodies comprising the heavy chain variable region (VH) and light chain variable region (VL) described herein.

[0008] The present disclosure also provides pharmaceutical compositions comprising a TF-ADC comprising an antibody or fragment thereof that binds to TF (a "TF antibody") and a drug conjugated (directly or indirectly) thereto. In some embodiments, such pharmaceutical compositions comprise a TF-ADC comprising an antibody or fragment thereof that binds to essentially the same epitope of human TF as an antibody comprising a VH and VL described herein.

[0009] The disclosure also provides methods of treating, preventing, or ameliorating a TF-mediated disease, disorder, or condition (e.g., a method of alleviating one or more symptoms of a TF-mediated disease, disorder, or condition using a TF-ADC).

[0010] More specifically, the disclosure provides TF-ADCs comprising (a) a TF antibody and (b) one or more pyridazine-pyrrolo coupling moieties comprising a drug conjugated to the pyridazine-pyrrolo coupling moiety via a linker, e.g., using hydrazino-iso-Picteth-Spengler (HIPS) conjugation methods.

[0011] Traditionally, HIPS conjugation methods have been used to generate conjugates with one payload per HIPS moiety per aldehyde tag, resulting in antibody conjugates with DAR values ​​of up to 4. In some embodiments, the TF-ADCs disclosed herein comprise a branched HIPS linker with two (or more) of the same or different payload molecules per HIPS moiety, thus enabling the conjugation of two (or more) small molecule payloads per aldehyde group in a protein in a single conjugation step (Figure 2). Consequently, the use of such branched linkers allows for the generation of higher DAR site-specific conjugates (e.g., DARs up to 8) with controlled payload placement, which, in the context of therapeutic ADCs, may result in higher amounts of pharmaceutical agent delivered to target tissues.

[0012] The present disclosure provides TF-ADC structures comprising (a) a TF antibody, (b) a branched HIPS linker, and (c) a drug. The disclosure also encompasses compounds and methods for producing such conjugates, as well as methods of using the conjugates.

[0013] Embodiments of the disclosure include TF-ADCs comprising: (a) a TF antibody; and (b) one or more pyridazine-pyrrolo coupling moieties that comprise one or more drugs conjugated to the pyridazine-pyrrolo coupling moieties via one or more linkers.

[0014] In some embodiments, the TF-ADC is represented by formula (I), and the TF-ADC comprises: a. an antibody that binds to tissue factor (TF), and b. Two or more drugs, each conjugated via a linker to a pyridazine-pyrrolo coupling moiety; [ka] (In the formula, Ab represents an antibody that binds to TF; Z 1 , Z 2 , and Z 4 However, each independently, CR 4 and Z 3 But CL B -W 2 and R 1 , R 2 , R 3 , and R 4 are each selected from hydrogen and alkyl; L A is a first linker comprising: -(T 1 -V 1 ) a -(T 2 -V 2 ) b -(T 3 -V 3 ) c -(T 4 -V4 ) d -(T 5 -V 5 ) e -(T 6 -V 6 ) f -, 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; T 1 , T 2 , T 3 , T 4 , T 5 , and T 6 are each independently 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-aminopiperidine (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 group, hydrazine, disulfide, and ester; EDA is an ethylenediamine moiety; PEG is polyethylene glycol; AA is an amino acid residue or amino acid analog; each w is an integer of 1 to 20; each n is an integer of 1 to 30; each p is an integer of 1 to 20; and each x is an integer of 1 to 12; V 1 , V 2 , V 3 , V 4 , V 5 , and V 6 are each independently a covalent bond, -CO-, or -NR 15 -, -NR15 (CH2) q -, -NR 15 (C6H4)-, -CONR 15 -, -NR 15 CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR 15 -, -NR 15 and -SO2-, and -P(O)OH-, 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 are 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 is the second linker: -(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 -, g, h, i, j, k, l, and m are each independently 0 or 1, and at least one of g, h, i, j, k, l, and m is 1; T 7 , T 8 , T 9 , T 10 , T11 , T 12 , and T 13 are each independently 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-aminopiperidine (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 group, hydrazine, disulfide, and ester; EDA is an ethylenediamine moiety; PEG is polyethylene glycol; AA is an amino acid residue or amino acid analog; each w is an integer of 1 to 20; each n is an integer of 1 to 30; each p is an integer of 1 to 20; and each x is an integer of 1 to 12; V 7 , V 8 , V 9 , V 10 , V 11 , V 12 , and V 13 are each independently a covalent bond, -CO-, or -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 and -SO2-, and -P(O)OH-, where each q is an integer from 1 to 6; Each R13 is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl; Each R 15 are 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, W 1 is the first drug, W 2 is the second drug).

[0015] In some embodiments, Z 1 But, CR 4 is.

[0016] In some embodiments, Z 3 But CL B -W 2 is.

[0017] In some embodiments, W 1 and W 2 One or both of the is a camptothecin analog (e.g., belotecan).

[0018] In some embodiments, L A but includes: -(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, 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; T 1 , T 2 , T 3 , T 4 , T 5 , and T 6 are each independently 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-aminopiperidine (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 group, hydrazine, disulfide, and ester; EDA is an ethylenediamine moiety; PEG is polyethylene glycol; AA is an amino acid residue or amino acid analog; each w is an integer of 1 to 20; each n is an integer of 1 to 30; each p is an integer of 1 to 20; and each x is an integer of 1 to 12; V 1 , V 2 , V 3 , V 4 , V 5 , and V 6 are each independently a covalent bond, -CO-, or -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 and -SO2-, and -P(O)OH-, 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 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.

[0019] L A In some embodiments of T 1 However, (C1-C 12 ) alkyl and substituted (C1-C 12 ) alkyl; T 2 , T 3 , T 4 , T 5 , and T 6 are each independently 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-aminopiperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, acetal groups, hydrazines, and esters; V 1 , V 2 , V 3 , V 4 , V 5 , and V6 are each independently a covalent bond, -CO-, or -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 selected from the group consisting of SO2-, and -P(O)OH-; (PEG) n but, [ka] and n is an integer from 1 to 30; EDA is an ethylenediamine moiety having the following structure: [ka] y is an integer of 1 to 6, and r is an integer of 0 to 1; 4-aminopiperidine (4AP) [ka] and Each R 12 are independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, and any two adjacent R 12 The groups may be cyclically linked to form a piperazinyl ring. In a further embodiment, a, b, c, and d are each 1; e and f are 0.

[0020] In some embodiments, T 1 , T 2 , T 3 , T 4 , T 5 , and T 6 are each optionally substituted with a glycoside.

[0021] In some embodiments, MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP are each optionally substituted with a glycoside.

[0022] In some embodiments, the glycoside is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.

[0023] In some embodiments, L A is the linker, T 1 However, (C1-C 12 ) alkyl, and V 1 is -CONH-, T 2 is replaced by (C1-C 12 ) alkyl, and V 2 is -CO-, T 3 However, (AA) p and V 3 But it doesn't exist, T 4 But PABC, V 4 But it doesn't exist, p is an integer from 1 to 10, a, b, c, and d are each 1; e and f are each 0.

[0024] In further embodiments, the PABC is substituted with a glycoside, eg, a hydrogen of the PABC is replaced with a glycoside (eg, glucuronide, galactoside, glucoside, mannoside, fucoside, O-GlcNAc, and O-GalNAc).

[0025] In some embodiments, L B but includes: -(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 -, g, h, i, j, k, 1, and m are each independently 0 or 1, provided that at least one of g, h, i, j, k, 1, and m is 1; T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 are each independently 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-aminopiperidine (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 group, hydrazine, disulfide, and ester; EDA is an ethylenediamine moiety; PEG is polyethylene glycol; AA is an amino acid residue or amino acid analog; each w is an integer of 1 to 20; each n is an integer of 1 to 30; each p is an integer of 1 to 20; and each x is an integer of 1 to 12; V 7 , V8 , V 9 , V 10 , V 11 , V 12 , and V 13 are each independently a covalent bond, -CO-, or -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 and -SO2-, and -P(O)OH-, 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 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.

[0026] In some embodiments, T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 are each optionally substituted with a glycoside.

[0027] In some embodiments, MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP are each optionally substituted with a glycoside.

[0028] In some embodiments, the glycoside is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.

[0029] L B In some embodiments of T 7 is a covalent bond, T 8 , T 9 , T 10 , T 11 , and T 12 are each independently 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-aminopiperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, acetal groups, hydrazines, and esters; V 7 , V 8 , V 9 , V 10 , V 11 , and V 12 are each independently a covalent bond, -CO-, or -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 SO2-, and -P(O)OH-; (PEG) n but, [ka] and n is an integer from 1 to 30; EDA is an ethylenediamine moiety having the following structure: [ka] y is an integer of 1 to 6, and r is an integer of 0 to 1; 4-aminopiperidine (4AP) [ka] and Each R 12 are independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, and any two adjacent R 12 The groups may be cyclically linked to form a piperazinyl ring, g, h, i, j, and k are each 1; l and m are 0.

[0030] 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 are each optionally substituted with a glycoside.

[0031] In some embodiments, MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP are each optionally substituted with a glycoside.

[0032] In some embodiments, the glycoside is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.

[0033] In some embodiments, L B is the linker, T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CONH-, T 9 is replaced by (C1-C 12 ) alkyl, and V 9 is -CO-, T 10 However, (AA) p and V 10 But it doesn't exist, T 11 But PABC, V 11 But it doesn't exist, p is an integer from 1 to 10, g, h, i, j, and k are each 1; l and m are each 0.

[0034] In further embodiments, the PABC is substituted with a glycoside, eg, a hydrogen of the PABC is replaced with a glycoside (eg, glucuronide, galactoside, glucoside, mannoside, fucoside, O-GlcNAc, and O-GalNAc).

[0035] In some embodiments, the TF-ADC has formula (I): [ka] (In the formula, Ab represents an antibody that binds to TF; Z 1 , Z 2 , and Z 4 However, each independently, CR 4 and Z 3 But CL B -W 2 and R1 , R 2 , R 3 , and R 4 are hydrogen and (C1-C 12 ) alkyl; L A is the first linker, T 1 However, (C1-C 12 ) alkyl, and V 1 is -CONH-, T 2 is replaced by (C1-C 12 ) alkyl, and V 2 is -CO-, T 3 However, (AA) p where p is an integer from 1 to 20, and V 3 is a covalent bond, T 4 But PABC, V 4 is a covalent bond, a, b, c, and d are each 1; e and f are each 0; L B is the second linker, T 7 is a covalent bond, and V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CONH-, T 9 is replaced by (C1-C 12 ) alkyl, and V 9 is -CO-, T 10 However, (AA) p where p is an integer from 1 to 20, and V 10 is a covalent bond, T 11 But PABC, V 11 is a covalent bond, g, h, i, j, and k are each 1; l and m are each 0; s is an integer from 1 to 10, W 1 is the first drug, W 2 is the second drug).

[0036] In some embodiments, W 1 and W 2 One or both of the is a camptothecin analog (e.g., belotecan).

[0037] In some embodiments, the TF-ADC has formula (I): [ka] (In the formula, Ab represents an antibody that binds to TF; Z 1 , Z 2 , and Z 4 However, each independently, CR 4 and Z 3 But CL B -W 2 and R 1 , R 2 , R 3 , and R 4 are hydrogen and (C1-C 12 ) alkyl; L A is the linker, T 1 is (C1-C6) alkyl, and V 1 is -CONH-, T 2 But -NHCO(PEG) k where k is an integer from 2 to 10, and V 2 is -CO-, T 3 is (AA)2 and V 3 is a covalent bond, T 4 is a glycosidic-substituted PABC, and V 4is a covalent bond, a, b, c, and d are each 1; e and f are each 0; L B is the linker, T 7 is a covalent bond, and V 7 is -NHCO-, T 8 is (C1-C6) alkyl, and V 8 is -CONH-, T 9 But -NHCO(PEG) k where k is an integer from 2 to 10, and V 9 is -CO-, T 10 is (AA)2 and V 10 is a covalent bond, T 11 is a glycosidic-substituted PABC, and V 11 is a covalent bond, g, h, i, j, and k are each 1; l and m are each 0; s is an integer from 1 to 10, W 1 is the first drug, W 2 is the second drug).

[0038] In some embodiments, T 4 and T 11 In some embodiments, one or both PABCs in T are substituted with a glucuronide. 1 and T 8 In some embodiments, one or both of T 2 and T 9 One or both of the groups may be -NHCO(PEG) k and k is an integer from 5 to 10. In some embodiments, W 1 and W 2One or both of the is a camptothecin analog (e.g., belotecan).

[0039] In some embodiments, the TF-ADC has formula (II): [ka] (In the formula, Ab represents an antibody that binds to TF; and s is an integer from 1 to 10.

[0040] In some embodiments, s is an integer from 1 to 4.

[0041] Formula (II) can be prepared by conjugating one or more linker payloads of formula (IIa) shown below to the TF antibody: [ka]

[0042] In some embodiments, the TF-ADC can be represented by formula (I) or (II), and the Ab comprises a VH CDR1, a VH CDR2, and a VH CDR3 set forth in a VH comprising the amino acid sequence of SEQ ID NO: 25, and a VL CDR1, a VL CDR2, and a VL CDR3 set forth in a VL comprising the amino acid sequence of SEQ ID NO: 26.

[0043] In some embodiments, the TF-ADC can be represented by formula (I) or (II), and the Ab comprises a VH CDR1, VH CDR2, and VH CDR3 set forth in a VH comprising the amino acid sequence of SEQ ID NO: 41, and a VL CDR1, VL CDR2, and VL CDR3 set forth in a VL comprising the amino acid sequence of SEQ ID NO: 42.

[0044] In some embodiments, a TF-ADC can be represented by formula (I) or (II), and the Ab comprises: (i) a VH region comprising: a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 1, 7, 8, 15, 21, 27, 31, 32, 35, or 39; a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 2, 9, 14, 16, or 22; and a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 3, 10, 17, or 23; and (ii) a VL region comprising: a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 4, 11, 18, 24, 28, 33, 36, or 40; a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 5, 12, 19, 29, or 37; and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 6, 13, 20, 30, 34, or 38.

[0045] In some embodiments, the TF-ADC can be represented by formula (I) or (II), wherein the Ab competes with any one of the TF antibodies disclosed herein for binding to TF.

[0046] In some embodiments, the TF-ADC can be represented by formula (I) or (II), and the Ab comprises framework 1 (FR1), framework 2 (FR2), framework 3 (FR3), and / or framework 4 (FR4) sequences set forth in any one of SEQ ID NOs: 25, 26, 41, and 42.

[0047] In some embodiments, the TF-ADC can be represented by formula (I) or (II), and the Ab comprises human framework sequences.

[0048] In some embodiments, the TF-ADC can be represented by formula (I) or (II), and the Ab comprises a VH comprising the amino acid sequence of SEQ ID NO: 25 and a VL comprising the amino acid sequence of SEQ ID NO: 26.

[0049] In some embodiments, a TF-ADC can be represented by formula (I) or (II), and the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 82. Additionally or alternatively, the Ab comprises a VL comprising the amino acid sequence of SEQ ID NO: 91. In some embodiments, a TF-ADC can be represented by formula (I) or (II), and the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 82 and a VL comprising the amino acid sequence of SEQ ID NO: 91.

[0050] In some embodiments, a TF-ADC can be represented by formula (I) or (II), and the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 79. Additionally or alternatively, the Ab comprises a VL comprising the amino acid sequence of SEQ ID NO: 91. In some embodiments, a TF-ADC can be represented by formula (I) or (II), and the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 79 and a VL comprising the amino acid sequence of SEQ ID NO: 91.

[0051] In some embodiments, the TF-ADC can be represented by formula (I) or (II), and the Ab comprises (i) a VH comprising the amino acid sequence of SEQ ID NO: 41, and a VL comprising the amino acid sequence of SEQ ID NO: 42.

[0052] In some embodiments, a TF-ADC can be represented by formula (I) or (II), and the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 85. Additionally or alternatively, the Ab comprises a VL comprising the amino acid sequence of SEQ ID NO: 92. In some embodiments, a TF-ADC can be represented by formula (I) or (II), and the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 85 and a VL comprising the amino acid sequence of SEQ ID NO: 92.

[0053] In some embodiments, a TF-ADC can be represented by formula (I) or (II), and the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 88. Additionally or alternatively, the Ab comprises a VL comprising the amino acid sequence of SEQ ID NO: 92. In some embodiments, a TF-ADC can be represented by formula (I) or (II), and the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 88 and a VL comprising the amino acid sequence of SEQ ID NO: 92.

[0054] The present disclosure also provides pharmaceutical compositions comprising a TF-ADC (the TF-ADC can be represented by Formula (I) or Formula (II)) and a pharmaceutically acceptable excipient (wherein the TF antibody (TF Ab or Ab) is as described in any embodiment described herein). In some embodiments, such pharmaceutical compositions may have a drug-to-antibody ratio (DAR) of the TF-ADC of about 1 to about 20, e.g., 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 7.5, about 7 to about 8, about 6.5, about 7, about 7.5, or about 8.

[0055] The disclosure also provides a method for treating cancer or tumor in a subject, the method comprising administering to the subject a TF-ADC (the TF-ADC can be represented by formula (I) or (II)), or a pharmaceutical composition comprising a TF-ADC of formula (I) or (II) and a pharmaceutically acceptable excipient, wherein the TF antibody is as described in any embodiment herein.

[0056] Provided herein is a kit comprising an antibody-drug conjugate disclosed herein or a pharmaceutical composition disclosed herein and instructions for use. [Brief explanation of the drawings]

[0057] [Figure 1]Schematic of HIPS ligation for ADC synthesis: An antibody bearing an aldehyde moiety is reacted with a hydrazino-iso-Picteth-Spengler (HIPS) linker and a payload to yield a site-specifically conjugated ADC bearing a stable azacarboline bond. [Figure 2]

[0023] Figure 1 shows a schematic diagram of branched HIPS ligation for ADC synthesis. An antibody with four aldehyde sites is reacted with a branched HIPS linker to yield ADCs with drug-to-antibody (DAR) values ​​of up to 8, according to embodiments of the present disclosure. [Figure 3A] Graphs of total antibody and ADC concentrations for pharmacokinetic (PK) sample analysis are shown. Squares: TF-ADC 6-8-total mAb measurement; Triangles: TF-ADC 6-8-total ADC measurement. [Figure 3B] Graphs of total antibody and ADC concentrations for pharmacokinetic (PK) sample analysis are shown. Diamonds: TF-ADC 6-4-total mAb measurement; Stars: TF-ADC 6-4-total ADC measurement. [Figure 3C] Graphs of total antibody and ADC concentrations for pharmacokinetic (PK) sample analysis are shown. Inverted triangles: TF-ADC 7-8 - total mAb measurement; crosses: TF-ADC 7-8 - total ADC measurement. [Figure 3D] Graphs of total antibody and ADC concentrations for pharmacokinetic (PK) sample analysis are shown. Stars: TF-ADC 7-4-total mAb measurement; Pluses: TF-ADC 7-4-total ADC measurement. [Figure 4] Graphs of in vitro cytotoxicity assays of free drugs (i.e., monomethyl auristatin E (MMAE), belotecan, or exatecan derivative DxD) in various TF-positive cancer cell lines (including RF / 6A, A431, HCT-116, BxPC3, AU-565, HPAF-II, MDA-MB-231, AsPC-1, HCC1954, and SKOV-3). [Figure 5]Graphs are shown of in vitro cytotoxicity assays of TF-ADC 6-8TF antibody conjugated to belotecan (gray circles), TF-ADC 7-8TF antibody conjugated to belotecan (squares), corresponding isotype antibody conjugated to belotecan (triangles), MMAE (diamonds), or free belotecan (black circles) in various TF-positive cancer cell lines (including RF / 6A, A431, HCT-116, BxPC3, AU-565, HPAF-II, MDA-MB-231, AsPC-1, HCC1954, and SKOV-3). [Figure 6] Graphs of in vitro cytotoxicity assays of FITC-ADC 8 (upper panel, circles marked with FITC), TF-ADC 6-8 (upper panel, unmarked squares), TF-ADC 7-8 (upper panel, triangles), belotecan (upper panel, squares marked with B), FITC-ADC 4 (lower panel, circles marked with FITC), TF-ADC 6-4 (lower panel, squares), TF-ADC 7-4 (lower panel, triangles), MMAE (lower panel, diamonds), or belotecan (lower panel, circles marked with B) in three selected TF-positive cancer cell lines (including A431, HPAF-II, and BxPC-3). [Figure 7] Figure 1 shows a graph of in vitro cytotoxicity assays of TF-ADC 6-8 with various DAR levels in three selected TF-positive cancer cell lines (including A431, HPAF-II, and BxPC-3). [Figure 8] Figure 1 shows a graph of in vitro cytotoxicity assay of TF-ADC 6-4 with different DAR levels in three selected TF-positive cancer cell lines (including A431, HPAF-II, and BxPC-3). [Figure 9A] A graph of mean tumor volume (mm3) versus days is shown, demonstrating the in vivo efficacy of TF-targeted ADCs against BxPC3 xenografts. [Figure 9B] The body weights of the mice tested are shown. [Figure 9C] In vivo efficacy data for TF-ADC 6-8 and TF-ADC 7-8 are plotted. [Figure 9D] In vivo efficacy data for TF-ADC 6-4 and TF-ADC 7-4 are plotted. [Figure 9E] In vivo efficacy data for TF-ADC 6-4 and TF-ADC 6-8 are plotted. [Figure 9F] In vivo efficacy data for TF-ADC 7-4 and TF-ADC 7-8 are plotted. A single iv dose was administered on Day 0. [Figure 10A] 1 shows a graph of the in vivo efficacy of TF-targeted ADCs against HPAF-II xenografts. [Figure 10B] The body weights of the mice tested are shown. [Figure 10C] In vivo efficacy data for TF-ADC 6-8 and TF-ADC 7-8 are plotted. [Figure 10D] In vivo efficacy data for TF-ADC 6-4 and TF-ADC 7-4 are plotted. [Figure 10E] In vivo efficacy data for TF-ADC 6-4 and TF-ADC 6-8 are plotted. [Figure 10F] Figure 10 plots in vivo efficacy data for TF-ADC 7-4 and TF-ADC 7-8. A single iv dose was administered on day 0. [Figure 11A] 1 shows a graph of the concentration of the indicated ADC (tADC) and its antibody in the plasma of mice administered various ADCs. Data are plotted for mice administered 3 mg / kg or 10 mg / kg of TF-ADC 6-4 or TF-ADC 6-8. [Figure 11B] Figure 11B shows graphs of the concentrations of the indicated ADC (tADC) and its antibody in the plasma of mice administered various ADCs. Data are plotted for mice administered 1 mg / kg of TF-ADC 6-4 or TF-ADC 6-8. As shown in Figure 11B, EXMA-006 serves as the negative control in both graphs, while the benchmark ADC served as a comparison control. [Figure 12A]12A and 12B show graphs of tumor volumes measured in HPAF-II xenografted mice treated with various dosages of TF-ADC 6-4 (FIG. 12A) or TF-ADC 6-8 (FIG. 12B). [Figure 12B] 12A and 12B show graphs of tumor volumes measured in HPAF-II xenografted mice treated with various dosages of TF-ADC 6-4 (FIG. 12A) or TF-ADC 6-8 (FIG. 12B). [Figure 13A] Presented are concentrations of the indicated ADCs (ADCs) and their antibodies (tAbs) in the plasma of non-human primates (NHPs) treated with various dosages of TF-ADC 6-4 (FIG. 13A) or TF-ADC 6-8 (FIG. 13B). [Figure 13B] Presented are concentrations of the indicated ADCs (ADCs) and their antibodies (tAbs) in the plasma of non-human primates (NHPs) treated with various dosages of TF-ADC 6-4 (FIG. 13A) or TF-ADC 6-8 (FIG. 13B). [Figure 14A] Payload concentrations in the plasma of NHPs treated with various dosages of TF-ADC 6-4 (Figure 14A) or TF-ADC 6-8 (Figure 14B) are presented. [Figure 14B] Payload concentrations in the plasma of NHPs treated with various dosages of TF-ADC 6-4 (Figure 14A) or TF-ADC 6-8 (Figure 14B) are presented. [Figure 15A] Exemplary ATP release results for treated tumor cells are presented (Figures 15A-15B, A431; Figure 15C, SKOV3). AUC is plotted for treatment groups with control (isotype IgG1 antibody conjugated to belotecan), 33 nM TF-ADC 6-8, 33 nM belotecan, 100 nM MMAE, or 100 nM benchmark ADC. [Figure 15B] Exemplary ATP release results for treated tumor cells are presented (Figures 15A-15B, A431; Figure 15C, SKOV3). Area under the curve (AUC) for tumor cells treated with various concentrations of TF-ADC 6-8, isotype control (isotype IgG1 antibody conjugated to belotecan), or belotecan are presented. [Figure 15C] Exemplary ATP release results for treated tumor cells are presented (Figures 15A-15B, A431; Figure 15C, SKOV3). Area under the curve (AUC) for tumor cells treated with various concentrations of TF-ADC 6-8, isotype control (isotype IgG1 antibody conjugated to belotecan), or belotecan are presented. [Figure 16] HMGB1 released by A431 tumor cells treated with control, 33 nM TF-ADC 6-8, 33 nM belotecan, 100 nM MMAE, or 100 nM benchmark ADC is plotted. [Figure 17A] IFNγ released by PBMCs co-cultured with tumor cells pre-treated with isotype ADC, TF-ADC 6-8, free belotecan, or free MMAE and anti-PD-1 antibody (hereafter referred to as MMAE+aPD1) for 24 hours (FIG. 17A) or 96 hours (FIG. 17B) is plotted. Each bar represents data from one donor. [Figure 17B] IFNγ released by PBMCs co-cultured with tumor cells pre-treated with isotype ADC, TF-ADC 6-8, free belotecan, or free MMAE and anti-PD-1 antibody (hereafter referred to as MMAE+aPD1) for 24 hours (FIG. 17A) or 96 hours (FIG. 17B) is plotted. Each bar represents data from one donor. [Figure 18A] IFNα released by PBMCs co-cultured with tumor cells pretreated with isotype ADC, TF-ADC 6-8, belotecan, or MMAE and an anti-PD-1 antibody (hereafter referred to as MMAE+aPD1) for 24 hours (FIG. 18A) or 96 hours (FIG. 18B) is plotted. Each bar represents data from one donor. [Figure 18B] IFNα released by PBMCs co-cultured with tumor cells pretreated with isotype ADC, TF-ADC 6-8, belotecan, or MMAE and an anti-PD-1 antibody (hereafter referred to as MMAE+aPD1) for 24 hours (FIG. 18A) or 96 hours (FIG. 18B) is plotted. Each bar represents data from one donor. [Figure 19A] IP-10 released by PBMCs co-cultured with tumor cells pretreated with isotype ADC, TF-ADC 6-8, belotecan, or MMAE and an anti-PD-1 antibody (hereafter referred to as MMAE+aPD1) for 24 hours (FIG. 19A) or 96 hours (FIG. 19B) is plotted. Each bar represents data from one donor. [Figure 19B] IP-10 released by PBMCs co-cultured with tumor cells pretreated with isotype ADC, TF-ADC 6-8, belotecan, or MMAE and an anti-PD-1 antibody (hereafter referred to as MMAE+aPD1) for 24 hours (FIG. 19A) or 96 hours (FIG. 19B) is plotted. Each bar represents data from one donor. [Figure 20A] Figures 20A and 20B show plots of MIP-1α released by PBMCs co-cultured with tumor cells pretreated with isotype ADC, TF-ADC 6-8, belotecan, or MMAE and an anti-PD-1 antibody (hereafter referred to as MMAE+aPD1) for 24 hours (Figure 20A) or 96 hours (Figure 20B). Each bar represents data from one donor. [Figure 20B] Figures 20A and 20B show plots of MIP-1α released by PBMCs co-cultured with tumor cells pretreated with isotype ADC, TF-ADC 6-8, belotecan, or MMAE and an anti-PD-1 antibody (hereafter referred to as MMAE+aPD1) for 24 hours (Figure 20A) or 96 hours (Figure 20B). Each bar represents data from one donor. DETAILED DESCRIPTION OF THE INVENTION

[0058] The present disclosure provides antibody-drug conjugates (ADCs) that bind to TF and drugs conjugated thereto (directly or indirectly). Such TF-ADCs are useful in compositions and methods for treating, preventing, or ameliorating TF-mediated diseases, disorders, or conditions (e.g., one or more symptoms of a disease, disorder, or condition). TF-mediated diseases, disorders, and conditions include various cancers (including, but not limited to, any cancer in which tumor cells express or overexpress TF). In addition, TF-ADCs are useful for killing and / or eliminating tumor cells. The TF-ADCs described herein are useful in compositions and methods for treating cancer. definition

[0059] The following terms have the following meanings unless otherwise indicated: Any undefined terms have their art-recognized meanings.

[0060] "Alkyl" refers to a monovalent saturated aliphatic hydrocarbon group having 1 to 10 carbon atoms, e.g., 1 to 6 carbon atoms, or 1 to 5, or 1 to 4, or 1 to 3 carbon atoms. This term includes, by way of example, straight-chain 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-)).

[0061] The term "substituted alkyl" refers to an alkyl group, as defined herein, wherein one or more carbon atoms in the alkyl chain (except the C1 carbon atom) is replaced with a heteroatom (e.g., -O-, -N-, -S-, -S(O) n-(n is 0-2), -NR-(R is hydrogen or alkyl) and optionally substituted with alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, 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-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-aryl, -SO2-heteroaryl, and -NR a R b and R has 1 to 5 substituents selected from the group consisting of ’ and R ” may be the same or different and are selected from hydrogen, optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl, and heterocyclic.

[0062] "Alkylene" refers to a divalent aliphatic hydrocarbon group, preferably having 1 to 6, more preferably 1 to 3 carbon atoms, either straight or branched, which is optionally substituted with -O-, -NR 10 -, -NR 10 C(O)-, -C(O)NR 10 and the like. This term includes, by way of example, methylene (-CH-), ethylene (-CHCH-), n-propylene (-CHCHCH-), isopropylene (-CHCH(CH)-), (-C(CH)CHCH-), (-C(CH)CHC(O)-), (-C(CH)CHC(O)NH-), (-CH(CH)CH-), and the like.

[0063] "Substituted alkylene" refers to an alkylene group having 1 to 3 hydrogens replaced with a substituent as set forth for carbon in the definition of "substituted" below.

[0064] The term "alkane" refers to alkyl and alkylene groups as defined herein.

[0065] The terms "alkylaminoalkyl," "alkylaminoalkenyl," and "alkylaminoalkynyl" refer to the group R'NHR"- where R' is an alkyl group, as defined herein, and R" is an alkylene, alkenylene, or alkynylene group, as defined herein.

[0066] The terms "alkaryl" or "aralkyl" refer to the groups -alkylene-aryl and -substituted alkylene-aryl, where alkylene, substituted alkylene, and aryl are defined herein.

[0067] "Alkoxy" refers to the group -O-alkyl, where alkyl is as defined herein. Alkoxy includes, by way of example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, t-butoxy, sec-butoxy, n-pentoxy, and the like. The term "alkoxy" also refers to the groups alkenyl-O-, cycloalkyl-O-, cycloalkenyl-O-, and alkynyl-O-, where alkenyl, cycloalkyl, cycloalkenyl, and alkynyl are defined herein.

[0068] The term "substituted alkoxy" refers to the groups 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.

[0069] The term "alkoxyamino" refers to the group --NH-alkoxy, where alkoxy is defined herein.

[0070] The term "haloalkoxy" refers to an alkyl-O- group in which one or more hydrogen atoms on the alkyl group has been replaced with a halo group and includes, by way of example, groups such as trifluoromethoxy.

[0071] The term "haloalkyl" refers to a substituted alkyl group as described above, in which one or more hydrogen atoms on the alkyl group have been replaced with a halo group. Examples of such groups include, but are not limited to, fluoroalkyl groups (e.g., trifluoromethyl, difluoromethyl, trifluoroethyl, etc.).

[0072] The term "alkylalkoxy" refers to the groups -alkylene-O-alkyl, alkylene-O-substituted alkyl, substituted alkylene-O-alkyl, and substituted alkylene-O-substituted alkyl, where alkyl, substituted alkyl, alkylene, and substituted alkylene are defined herein.

[0073] The term "alkylthioalkoxy" refers to the groups -alkylene-S-alkyl, alkylene-S-substituted alkyl, substituted alkylene-S-alkyl, and substituted alkylene-S-substituted alkyl, where alkyl, substituted alkyl, alkylene, and substituted alkylene are defined herein.

[0074] "Alkenyl" refers to a straight or branched chain hydrocarbon group having 2 to 6 carbon atoms, preferably 2 to 4 carbon atoms, and at least one, preferably one or two, sites of double bond unsaturation. This term includes, by way of example, bivinyl, allyl, and but-3-en-1-yl. This term includes cis and trans isomers, or mixtures of these isomers.

[0075] The term "substituted alkenyl" refers to an alkenyl group, as defined herein, having one to five substitutions, or one to three substituents, selected from alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, 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-alkyl, -SO-substituted alkyl, -SO-aryl, and -SO-heteroaryl.

[0076] "Alkynyl" refers to a straight- or branched-chain monovalent hydrocarbon radical having two to six carbon atoms, preferably two to three carbon atoms, and having at least one, and preferably one or two, sites of triple bond unsaturation. Examples of such alkynyl radicals include acetylenyl (-C≡CH) and propargyl (-CHC≡CH).

[0077] The term "substituted alkynyl" refers to an alkynyl group, as defined herein, having one to five substitutions, or one to three substituents, including alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, 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, It is selected from -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl, and -SO2-heteroaryl.

[0078] "Alkynyloxy" refers to the group -O-alkynyl, where alkynyl is as defined herein. Alkynyloxy includes, by way of example, ethynyloxy, propynyloxy, and the like.

[0079] "Acyl" means 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)- , substituted heteroaryl-C(O)—, heterocyclyl-C(O)—, and substituted heterocyclyl-C(O)—, 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. For example, acyl includes the "acetyl" group CHC(O)—.

[0080] "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 C(O)-substituted heterocycle, R 20is 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, heterocycle, and substituted heterocycle are as defined herein.

[0081] The term "aminocarbonyl" or "aminoacyl" refers to -C(O)NR 21 R 22 R refers to the group 21 and R 22 is 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 optionally, together with the nitrogen to which it is bound, form a heterocyclic or substituted heterocyclic group, 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.

[0082] "Aminocarbonylamino" is -NR 21 C(O)NR 22 R 23 R refers to the group 21 , R 22 , and R 23 is independently selected from hydrogen, alkyl, aryl, or cycloalkyl, or two R groups join to form a heterocyclyl group.

[0083] The term "alkoxycarbonylamino" refers to the group -NRC(O)OR, where each R is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, or heterocyclyl, where alkyl, substituted alkyl, aryl, heteroaryl, and heterocyclyl are defined herein.

[0084] 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.

[0085] "Aminosulfonyl" is -SO2NR 21 R 22 R refers to the group 21 and R 22 is 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 optionally, together with the nitrogen to which it is attached, form a heterocyclic or substituted heterocyclic group, 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.

[0086] "Sulfonylamino" is -NR 21 SO2R 22 R refers to the group 21 and R 22is 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 optionally, together with the atoms to which they are attached, form a heterocyclic or substituted heterocyclic group, 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.

[0087] "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 condensed rings (examples of such aromatic ring systems include naphthyl, anthryl, and indanyl), which may or may not be aromatic, provided that the point of attachment is through an aromatic ring atom. This term includes, by way of example, phenyl and naphthyl. Unless otherwise limited by the definition of the aryl substituent, such aryl groups can be optionally substituted with one to five substituents, or one to three 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, azido, 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.

[0088] "Aryloxy" refers to the group -O-aryl, where aryl is as defined herein, that includes, by way of example, phenoxy, naphthoxy, and the like, which optionally includes optionally substituted aryl groups, also as defined herein.

[0089] "Amino" refers to the group -NH2.

[0090] The term "substituted amino" refers to the group -NRR, 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.

[0091] The term "azido" refers to the group -N3.

[0092] "Carboxyl," "carboxy," or "carboxylate" refers to -CO2H or salts thereof.

[0093] The terms "carboxyl ester" or "carboxy esters" 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, -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, 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.

[0094] "(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 cycloalkenyl, -OC(O)O-heteroaryl, -OC(O)O-substituted heteroaryl, -OC(O)O-heterocyclic, and -OC(O)O-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.

[0095] "Cyano" or "nitrile" refers to the group --CN.

[0096] "Cycloalkyl" refers to cyclic alkyl groups of 3 to 10 carbon atoms having single or multiple cyclic rings (e.g., fused, bridged, and spiro ring systems). Examples of suitable cycloalkyl groups include, for example, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like. Such cycloalkyl groups include, for example, single-ring structures (e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like) or multiple-ring structures (e.g., adamantanyl, and the like).

[0097] The term "substituted cycloalkyl" refers to a cycloalkyl group having one to five substituents or one to three substituents, which may be alkyl, substituted alkyl, alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, 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- Alkyl, It is selected from -SO2-substituted alkyl, -SO2-aryl and -SO2-heteroaryl.

[0098] "Cycloalkenyl" refers to a non-aromatic cyclic alkyl group of 3 to 10 carbon atoms having a single or multiple rings and having at least one double bond, preferably 1 to 2 double bonds.

[0099] The term "substituted cycloalkenyl" refers to a cycloalkenyl group having one to five substituents, or one to three substituents, selected from alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, 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.

[0100] "Cycloalkynyl" refers to a non-aromatic cycloalkyl group of 5 to 10 carbon atoms having mono- or polycyclic rings and having at least one triple bond.

[0101] "Cycloalkoxy" refers to -O-cycloalkyl.

[0102] "Cycloalkenyloxy" refers to -O-cycloalkenyl.

[0103] "Halo" or "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0104] "Hydroxy" or "hydroxyl" refers to the group --OH.

[0105] "Heteroaryl" refers to an aromatic group of 1 to 15 carbon atoms (e.g., 1 to 10 carbon atoms) and 1 to 10 heteroatoms selected from the group consisting of oxygen, nitrogen, and sulfur within the ring. Such heteroaryl groups can have a single ring (e.g., pyridinyl, imidazolyl, or furyl) or multiple condensed rings within the ring system (e.g., groups such as indolizinyl, quinolinyl, benzofuran, benzimidazolyl, or benzothienyl), wherein at least one ring within the ring system is aromatic. To satisfy valence requirements, the heteroatoms in such heteroaryl rings can be bonded to H or a substituent, such as an alkyl group or other substituent described herein, or can be unbonded. In certain embodiments, the nitrogen and / or sulfur ring atom(s) of a heteroaryl group are optionally oxidized to provide N-oxide (N→O), sulfinyl, or sulfonyl moieties. This term includes, by way of example, pyridinyl, pyrrolyl, indolyl, thiophenyl, and furanyl. Unless otherwise limited by the definition of the heteroaryl substituent, such heteroaryl groups can be optionally substituted with one to five substituents, or one to three 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, azido, 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, and trihalomethyl.

[0106] The term "heteroaralkyl" refers to the group -alkylene-heteroaryl, where alkylene and heteroaryl are defined herein. This term includes, by way of example, pyridylmethyl, pyridylethyl, indolylmethyl, and the like.

[0107] "Heteroaryloxy" refers to -O-heteroaryl.

[0108] "Heterocycle," "heterocyclic," "heterocycloalkyl," and "heterocyclyl" refer to saturated or unsaturated groups having a single ring or multiple fused rings, including fused-bridged and spiro ring systems, and having 3 to 20 ring atoms, including 1 to 10 heteroatoms. These ring atoms are selected from nitrogen, sulfur, or oxygen; in fused ring systems, one or more of the rings can be cycloalkyl, aryl, or heteroaryl, provided that the point of attachment is through a non-aromatic ring. In certain embodiments, the nitrogen and / or sulfur atoms of a heterocyclic group are optionally oxidized to provide N-oxide, -S(O)-, or -SO2- moieties. To satisfy valence requirements, the heteroatoms in such heterocyclic rings can be bonded to one or more H or one or more substituent(s) (e.g., alkyl groups or other substituents described herein), or can be unbonded.

[0109] Examples of heterocyclic and heteroaryl compounds include azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, dihydroindole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazoline, and the like. Examples of suitable amines 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, morpholinyl, thiomorpholinyl (also called thiamorpholinyl), 1,1-dioxothiomorpholinyl, piperidinyl, pyrrolidine, tetrahydrofuranyl, and the like.

[0110] Unless otherwise limited by the definition of a heterocyclic substituent, such heterocyclic groups can be optionally substituted with one to five, or one to three, substituents, including alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, 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- It is selected from alkyl, -SO2-substituted alkyl, -SO2-aryl, -SO2-heteroaryl, and fused heterocycle.

[0111] "Heterocyclyloxy" refers to the group --O-heterocyclyl.

[0112] The term "heterocyclylthio" refers to a heterocyclic -S- group.

[0113] The term "heterocyclene" refers to a diradical group formed from a heterocycle, as defined herein.

[0114] The term "hydroxyamino" refers to the group --NHOH.

[0115] "Nitro" refers to the -NO2 group.

[0116] "Oxo" refers to the atom (=O).

[0117] "Sulfonyl" refers to -SO2-alkyl, -SO2-substituted alkyl, -SO2-alkenyl, -SO2-substituted alkenyl, -SO2-cycloalkyl, -SO2-substituted cycloalkyl, -SO2-cycloalkenyl, Sulfonyl refers to -SO2-substituted cycloalkenyl, -SO2-aryl, -SO2-substituted aryl, -SO2-heteroaryl, -SO2-substituted heteroaryl, -SO2-heterocyclic, and -SO2-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. Sulfonyl, by way of example, includes methyl-SO2-, phenyl-SO2-, and 4-methylphenyl-SO2-.

[0118] "Sulfonyloxy" refers to the groups -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.

[0119] "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, and -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, 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.

[0120] The term "aminocarbonyloxy" refers to the group -OC(O)NRR, where each R is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, or heterocyclic, where alkyl, substituted alkyl, aryl, heteroaryl, and heterocyclic are defined herein.

[0121] "Thiol" refers to the group --SH.

[0122] The term "thioxo" or "thioketo" refers to the atom (=S).

[0123] The term "alkylthio" or "thioalkoxy" refers to an -S-alkyl group, where alkyl is as defined herein. In certain embodiments, sulfur can be oxidized to -S(O)-. Sulfoxides can exist as one or more stereoisomers.

[0124] The term "substituted thioalkoxy" refers to the group --S-substituted alkyl.

[0125] The term "thioaryloxy" refers to an aryl-S- group, where aryl is as defined herein, including optionally substituted aryl groups, also defined herein.

[0126] The term "thioheteroaryloxy" refers to the group heteroaryl-S-, where heteroaryl is as defined herein, including optionally substituted aryl, also defined herein.

[0127] The term "thioheterocyclooxy" refers to the group heterocyclyl-S-, where heterocyclyl is as defined herein, including optionally substituted heterocyclyl groups, also defined herein.

[0128] In the present disclosure, the term "substituted," when used to modify a particular group or radical, can also mean that one or more hydrogen atoms of the particular group or radical are each, independently of one another, replaced with the same or different substituents defined below.

[0129] In addition to the groups disclosed for individual terms herein, substituents (such as ═O, ═NR, etc.) to replace one or more hydrogens on a saturated carbon atom or radical of a given group are also included.70 , =N-OR 70 , =N2, or =S) is -R unless otherwise specified. 60 , halo, =O, -OR 70 , -SR 70 , -NR 80 R 80 , trihalomethyl, -CN, -OCN, -SCN, -NO, -NO2, =N2, -N3, -SO2R 70 , -SO2O - M + , -SO2OR 70 , -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 , -NR70 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 is selected from the group consisting of optionally substituted alkyl, cycloalkyl, heteroalkyl, heterocycloalkylalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; and each R 70 are independently hydrogen or R 60 and each R 80 But independently, R 70 Or alternatively, two R 80 together with the nitrogen atom to which they are attached form a 5-, 6-, or 7-membered heterocycloalkyl, which optionally contains 1 to 4 of the same or different additional heteroatoms selected from the group consisting of O, N, and S, of which N may have —H or C1-C3 alkyl substitution; and each M + is a counterion with a net single positive charge. + may independently be, for example, an alkali ion (e.g., K + , Na + , Li + ); ammonium ions (e.g., + N(R 60 ) 4); or alkaline earth ions (e.g., [Ca 2+ ] 0.5 , [Mg 2+ ] 0.5 , or [Ba 2+ ] 0.5) (the subscript 0.5 indicates that one of the counterions to such divalent alkaline earth ions is the ionized form of a compound of the invention, and another representative counterion (e.g., chloride), or a two-ionized compound disclosed herein, can serve as the counterion to such divalent alkaline earth ion, or a doubly-ionized compound of the invention can serve as the counterion to such divalent alkaline earth ion). Specific examples include -NR 80 R 80 is meant to include -NH2, -NH-alkyl, N-pyrrolidinyl, N-piperazinyl, N-methylpiperazin-1-yl, and N-morpholinyl.

[0130] Further to the disclosure herein, the substitution of hydrogen on an unsaturated carbon atom in a "substituted" alkene, alkyne, aryl, and heteroaryl group is represented by -R unless otherwise specified. 60 , halo, -O - M + , -OR 70 , -SR 70 , -S - M + , -NR 80 R 80 , trihalomethyl, -CF3, -CN, -OCN, -SCN, -NO, -NO2, -N3, -SO2R 70 , -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 R80 , -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)R 70 , -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 + is as defined above, except that in the case of a substituted alkene or alkyne, the substituent is -O - M + , -OR 70 , -SR 70 , or -S - M + isn't it.

[0131] In addition to the groups disclosed herein for each individual term, the substituent of a hydrogen on a nitrogen atom of a "substituted" heteroalkyl and cycloheteroalkyl group is, 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 + is as defined above.

[0132] Further to the disclosure herein, in certain embodiments, a substituted group has 1, 2, 3, or 4 substituents, 1, 2, or 3 substituents, 1 or 2 substituents, or 1 substituent.

[0133] For all substituents defined above, it is understood that polymers obtained by substituting the substituent itself with a further substituent (e.g., a substituted aryl having a substituted aryl group as a substituent, which is itself substituted with a substituted aryl group) are not intended to be included herein. In such cases, the maximum number of substitutions is 3. For example, the sequential substitution of substituted aryl groups specifically contemplated herein is limited to substituted aryl-(substituted aryl)-substituted aryl.

[0134] Unless otherwise indicated, the nomenclature of substituents not explicitly defined herein is performed by naming the terminal portion of the functional group followed by the adjacent functional group toward the point of attachment. For example, the substituent "arylalkyloxycarbonyl" refers to the group (aryl)-(alkyl)-OC(O)-.

[0135] For any of the groups disclosed herein that contain one or more substituents, it is of course understood that such groups do not contain any substitutions or substitution patterns that are sterically impractical and / or synthetically impractical. Additionally, the subject compounds include all stereochemical isomers arising from the substitution of these compounds.

[0136] The term "pharmaceutically acceptable salt" refers to a salt that is acceptable for administration to a patient (e.g., a mammal) (i.e., a salt containing a counterion that has acceptable mammalian safety 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 pharmaceutically acceptable salts of a compound, which are derived from a variety of organic and inorganic counterions well known in the art, including, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, etc., and, if the molecule contains a basic functional group, includes salts of organic or inorganic acids (e.g., hydrochloride, hydrobromide, formate, tartrate, besylate, mesylate, acetate, maleate, oxalate, etc.).

[0137] The term "salt thereof" refers to a compound formed when a proton of an acid is replaced by a cation (e.g., a metal cation or an organic cation, etc.). Where applicable, the salt is a pharmaceutically acceptable salt, although this is not required for salts of intermediate compounds not intended for administration to a patient. By way of example, salts of the present compounds include those that are protonated with an inorganic or organic acid to form a cation, with the conjugate base of the inorganic or organic acid as the anionic component of the salt.

[0138] A "solvate" refers to a complex formed by the combination of solvent molecules with solute molecules or ions. The solvent may be 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.

[0139] "Stereoisomer" and "stereoisomers" refer to compounds that have the same atomic bonds but different atomic arrangements in space. Stereoisomers include cis-trans isomers, E isomers, Z isomers, enantiomers, and diastereomers.

[0140] "Tautomers" refer to alternative forms of molecules that differ only in the position of the atoms' electron bonding and / or protons (e.g., enol-keto and imine-enamine tautomers), or tautomeric forms of heteroaryl groups (e.g., containing the -N=C(H)-NH- ring atom configuration (e.g., pyrazole, imidazole, benzimidazole, triazole, tetrazole)). One of ordinary skill in the art will recognize that other tautomeric ring atom configurations are possible.

[0141] It will be understood that the term "or a salt or solvate or stereoisomer thereof" is intended to include all permutations of salts, solvates, and stereoisomers (e.g., solvates of pharmaceutically acceptable salts of stereoisomers of the subject compound).

[0142] "Pharmaceutically effective amount" and "therapeutically effective amount" refer to an amount of a compound sufficient to treat a particular disorder or disease, or one or more of its symptoms, and / or prevent the onset of the disease or disorder. With respect to tumorigenic proliferative diseases, a pharmaceutically or therapeutically effective amount includes, inter alia, an amount sufficient to shrink a tumor or reduce the rate of tumor growth.

[0143] "Patient" refers to human and non-human subjects, particularly mammalian subjects.

[0144] As used herein, the term "treating" or "treatment" means treating or managing a disease or condition in a patient (e.g., a mammal, particularly a human), including: (a) preventing the occurrence of the disease or condition in question, e.g., prophylactic treatment; (b) ameliorating the disease or condition in the patient, e.g., ameliorating or causing regression of the disease or condition; (c) inhibiting the disease or condition in the patient, e.g., slowing or arresting the progression of the disease or condition; or (d) alleviating the symptoms of the disease or condition in the patient.

[0145] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to polymeric forms of amino acids of any length. Unless otherwise indicated, "polypeptide," "peptide," and "protein" can include genetically encoded and non-encoded amino acids, chemically or biochemically modified or derivatized amino acids, and polypeptides with modified peptide backbones. The terms include fusion proteins (including, but not limited to, fusion proteins with heterologous amino acid sequences, fusions with heterologous and homologous leader sequences, proteins containing at least one N-terminal methionine residue (e.g., to facilitate production in recombinant host cells), immunologically tagged proteins, etc.). In certain embodiments, the polypeptide is an antibody.

[0146] "Native amino acid sequence" or "parent amino acid sequence" are used interchangeably herein to refer to the amino acid sequence of a polypeptide before it has been modified to contain at least one modified amino acid residue.

[0147] The terms "amino acid analog," "unnatural amino acid," and the like, may be used interchangeably and include amino acid-like compounds similar in structure and / or overall 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, 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-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) are one or more modified groups within the molecule. Such modifications may include, but are not limited to, substituting an atom (e.g., N) for a related atom (e.g., S), adding a group (e.g., methyl, hydroxyl, etc.) or atom (e.g., Cl, Br, etc.), deleting a group, substituting a covalent bond (e.g., a single bond for a double bond), or a combination thereof. For example, amino acid analogs may include alpha-hydroxy acids, alpha-amino acids, and the like. Examples of amino acid analogs include, but are not limited to, sulfoalanine, and the like.

[0148] Terms such as "amino acid side chain" or "side chain of an amino acid" can be used to refer to a substituent attached to the alpha carbon of an amino acid residue (including natural amino acids, unnatural amino acids, and amino acid analogs). Amino acid side chains can also include amino acid side chains described in the context of modified amino acids and / or conjugates described herein.

[0149] Terms such as "carbohydrate" can be used to refer to monomeric units and / or polymers of monosaccharides, disaccharides, oligosaccharides, and polysaccharides. The term sugar can be 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 carbohydrate of interest 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 compounds and conjugates.

[0150] The term "glycoside" or "glycosyl" refers to a sugar molecule or group that is attached to a moiety via a glycosidic bond. For example, the moiety to which the glycoside is attached can be a cleavable linker described herein. The glycosidic bond can connect the glycoside to another moiety via various types of bonds, including, but not limited to, an O-glycosidic bond (O-glycoside), an N-glycosidic bond (glycosylamine), an S-glycosidic bond (thioglycoside), or a C-glycosidic bond (C-glycoside or C-glycosyl). In some cases, the glycoside can be cleaved from the moiety to which it is attached, for example, by chemical- or enzymatic-mediated hydrolysis.

[0151] The term "antibody" is used in the broadest sense and includes monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and multispecific antibodies (e.g., bispecific antibodies), humanized antibodies, single-chain antibodies, chimeric antibodies, and antibody fragments (e.g., Fab fragments). Antibodies are capable of binding to target antigens. (Janeway, C., Travers, P., Walport, M., Shlomchik (2001) Immuno Biology, 5th Ed., Garland Publishing, New York). Target antigens have one or more binding sites (also called epitopes), which are recognized by complementarity-determining regions (CDRs) formed by one or more variable regions of the antibody.

[0152] The term "natural antibody" refers to an antibody in which the heavy and light chains of the antibody are paired and produced by the immune system of a multicellular organism. The spleen, lymph nodes, bone marrow, and serum are examples of tissues that produce natural antibodies. For example, antibodies produced by antibody-producing cells isolated from a first animal immunized with an antigen are natural antibodies.

[0153] The term "humanized antibody" or "humanized immunoglobulin" refers to a non-human (e.g., mouse or rabbit) antibody that contains one or more amino acids (e.g., in a framework region, constant region, or CDR) substituted with an amino acid at the corresponding position in a human antibody. Generally, a humanized antibody elicits a reduced immune response in a human host compared to a non-humanized version of the same antibody. Antibodies can be humanized using various techniques known in the art, including, for example, CDR grafting (EP 239,400; PCT Publication WO 91 / 09967; U.S. Patent Nos. 5,225,539, 5,530,101, and 5,585,089), veneering or resurfacing (EP 592,106; EP 519,596; 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 No. 5,565,332). In certain embodiments, framework substitutions are identified by modeling CDR and framework residue interactions to identify framework residues important for antigen binding, and sequence comparison to identify unusual framework residues at specific positions (see, e.g., U.S. Pat. No. 5,585,089; Riechmann et al., Nature 332:323 (1988)). Further methods for humanizing antibodies contemplated for use in the present invention are described in 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 Publication Nos. WO98 / 45331 and WO98 / 45332. In certain embodiments, the subject rabbit antibodies can be humanized according to the methods described in US20040086979 and US20050033031. Thus, the above antibodies can be humanized using methods well known in the art.

[0154] The term "chimeric antibody" refers to an antibody whose light and heavy chain genes have been constructed, usually by genetic engineering, from variable and constant region genes of antibodies belonging to different species. For example, the variable segments of a mouse monoclonal antibody gene may be joined to human constant segments (e.g., gamma 1 and gamma 3). An example of a therapeutic chimeric antibody is a hybrid protein consisting of the variable or antigen-binding domain of a mouse antibody and the constant or effector domain of a human antibody, although domains from other mammalian species may be used.

[0155] The immunoglobulin light or heavy chain variable region of an immunoglobulin polypeptide is composed of a framework region (FR) flanked by three hypervariable regions, also called "complementarity-determining regions" or "CDRs." The extent of the framework region and CDRs has been defined (see, for example, "Sequences of Proteins of Immunological Interest," E. Kabat et al., USDapartment of Health and Human Services, 1991). The framework region of an antibody is the combined framework region of the constituent light and heavy chains and serves to position and align the CDRs. The CDRs are primarily responsible for binding to an epitope of an antigen.

[0156] A "parent Ig polypeptide" is a polypeptide comprising an amino acid sequence lacking an aldehyde-tagged constant region as described herein. The parent polypeptide may comprise a native sequence constant region or may comprise a constant region with pre-existing amino acid sequence modifications (e.g., additions, deletions, and / or substitutions).

[0157] As used herein, the term "isolated" is meant to describe a compound of interest that is present in an environment that is different from the environment in which the compound naturally occurs. "Isolated" means that the compound of interest is in a sample that is substantially enriched and / or includes compounds from which the compound of interest is partially or substantially purified.

[0158] As used herein, the term "substantially purified" refers to a compound that has been removed from its natural environment and is at least 60% free, at least 75% free, at least 80% free, at least 85% free, at least 90% free, at least 95% free, at least 98% free, or greater than 98% free from other components with which it is naturally associated.

[0159] The term "physiological conditions" is meant to encompass conditions compatible with living cells, eg, primarily aqueous conditions of temperature, pH, salinity, etc., compatible with living cells.

[0160] "Reaction partner" refers to a molecule or molecular moiety that specifically reacts with another reaction partner to produce a reaction product. Exemplary reaction partners include the cysteine ​​or serine of a sulfatase motif and formylglycine generating enzyme (FGE), which react to form a reaction product of a converted aldehyde tag that contains formylglycine (fGly) in place of the cysteine ​​or serine in the motif. Another exemplary reaction partner includes the aldehyde (e.g., reactive aldehyde group) of the fGly residue of a converted aldehyde tag and an "aldehyde-reactive reaction partner," which includes an aldehyde-reactive group and a moiety of interest, which reacts to form a reaction product of a polypeptide having a moiety of interest conjugated to the polypeptide via the fGly residue.

[0161] "N-terminus" refers to the terminal amino acid residue of a polypeptide having a free amine group; the amine group of the non-N-terminal amino acid residue usually forms part of the covalent backbone of the polypeptide.

[0162] "C-terminus" refers to the terminal amino acid residue of a polypeptide having a free carboxyl group; the carboxyl group of the non-C-terminal amino acid residue usually forms part of the covalent backbone of the polypeptide.

[0163] An "internal site" when used in reference to a polypeptide or an amino acid sequence of a polypeptide means a region of the polypeptide that is not at the N-terminus or C-terminus.

[0164] The term "subject" refers to human and non-human subjects, particularly mammalian subjects.

[0165] As used herein, the term "treating" or "treatment" means treating or managing a disease or condition in a subject (e.g., a mammal (particularly a human)), including: (a) preventing the occurrence of the disease or condition in a subject, e.g., prophylactic treatment; (b) ameliorating the disease or condition in a subject, e.g., ameliorating or causing regression of the disease or condition; (c) inhibiting the disease or condition in a subject, e.g., slowing or arresting the progression of the disease or condition; or (d) alleviating the symptoms of the disease or condition in a subject. In some embodiments, the term "treating" or "treatment" excludes prophylactic treatment.

[0166] As used herein, the term "native amino acid sequence" refers to the amino acid sequence of a polypeptide before it has been modified to contain an altered amino acid residue.

[0167] The terms "amino acid analog," "unnatural amino acid," and the like are used interchangeably and include amino acid-like compounds similar in structure and / or overall 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, 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-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) are one or more modified groups within the molecule. Such modifications can include, but are not limited to, substitution of an atom (e.g., N) for a related atom (e.g., S), addition of a group (e.g., methyl, hydroxyl, etc.) or atom (e.g., Cl, Br, etc.), deletion of a group, replacement of a covalent bond (such as a single bond for a double bond), or combinations thereof. For example, amino acid analogs include α-hydroxy acids, α-amino acids, etc.

[0168] The term "amino acid side chain" is used to refer to a substituent attached to the alpha carbon of an amino acid residue (including, for example, natural amino acids, unnatural amino acids, and amino acid analogs). Amino acid side chains can also include those amino acid side chains described in the context of modified amino acids and / or conjugates described herein.

[0169] 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 carbohydrate of interest 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 compounds and conjugates.

[0170] The term "glycoside" or "glycosyl" refers to a sugar molecule or group that is attached to a moiety via a glycosidic bond. For example, the moiety to which the glycoside is attached can be a cleavable linker described herein. The glycosidic bond can connect the glycoside to another moiety via various types of bonds, including, but not limited to, an O-glycosidic bond (O-glycoside), an N-glycosidic bond (glycosylamine), an S-glycosidic bond (thioglycoside), or a C-glycosidic bond (C-glycoside or C-glycosyl). In some cases, the glycoside can be cleaved from the moiety to which it is attached, for example, by chemical- or enzymatic-mediated hydrolysis.

[0171] The terms "tissue factor," "TF," "platelet tissue factor," "factor III," "thromboplastin," and "CD142" are used interchangeably herein to refer to TF, or any variants (e.g., splice and allelic variants), isoforms, and species homologs of TF, which are naturally expressed by a cell or expressed by a cell transfected with the TF gene. In some aspects, the TF protein is a TF protein naturally expressed by a primate (e.g., monkey or human), rodent (e.g., mouse or rat), dog, camel, cat, cow, goat, horse, pig, or sheep. The term TF encompasses "full-length" TF and any form of TF or fragment thereof (e.g., generated by intracellular processing). In some embodiments, TF includes a signal sequence. In some embodiments, TF does not include a signal sequence. In some embodiments, the term TF refers to a fragment of full-length TF that includes the TF extracellular domain (ECD). The term TF also encompasses naturally occurring variants of TF (e.g., SNP variants, splice variants, and allelic variants). In some embodiments, the TF protein is human TF (hTF; [ka] The underline indicates the signal peptide. In further embodiments, human TF does not include the signal peptide (e.g., amino acids (aa) 33 to 295 of SEQ ID NO: 175. In yet another embodiment, human TF as used herein refers to the extracellular domain (ECD) of human TF, e.g., amino acids 33 to 251 of SEQ ID NO: 175. In some aspects, the TF protein is derived from cynomolgus monkey TF (cTF; SGTTNTVAAYNLTWKSTNFKTILEWEPKPINQVYTVQISTKSGDWKSKCFYTADTECDLTDEIVKDVKQTYLARVFSYPAGHVESTGSTEEPPYENSPEFTPYLETNLGQPTIQSFEQVGTKVNVTVQDEWTLVRRNDTFLSLRDVFGKDLIYTLYYWKSSSSGKKTAKTNTNEFLIDVDKGENYCFSVQAVIPSRRTANRKSTDSPVECMGHEKGESREIFYIIGAVVFVVIILVIILAISLHKCKKARVGRSWKENSPLNVA (SEQ ID NO: 176). In further embodiments, cynomolgus TF as used herein refers to its ECD, e.g., aa 1 to aa 220 of SEQ ID NO: 176. In some aspects, the TF protein is mouse TF (mTF; AGIPEKAFNLTWISTDFKTILEWQPKPTNYTYTVQISDRSRNWKNKCFSTTDTECDLTDEIVKDVTWAYEAKVLSVPRRNSVHGDGDQLVIHGEEPPFTNAPKFLPYRDTNLGQPVIQQFEQDGRKLNVVVKDSLTLVRKNGTFLTLRQVFGKDLGYIITYRKGSSTGKKTNITNTNEFSIDVEEGVSYCFFVQAMIFSRKTNQNSPGSSTVCTEQWKSFLGETLIIVGAVVLLATIFIILLSISLCKRRKNRAGQKGKNTPSRLA (SEQ ID NO: 177). In further embodiments, mouse TF as used herein refers to its ECD, e.g., aa 1 to aa 223 of SEQ ID NO: 177. In some aspects, the TF protein is porcine TF (pTF; TGTTDVIVAYNLTWKSTNFKTILEWEPKPINYVYTVQISPRLGDWKNKCFHTTDTECDVTDEIMRNVKETYVARVLSYPADTVLTAQEPPFTNSPPFTPYLDTNLGQPVIQSFEQVGTKLNVTVEAARTLVRVNGTFLRLRDVFGKDLNYTLYYWRASSTGKKKATTNTNEFLIDVDKGENYCFSVQAVIPSRRVNQKSPESRIECTSQEKAVSRELFLIVGAVVFAVIVFVLVLSVSLYKCRKERAGPSGKENAPLNVA (SEQ ID NO: 178). In some embodiments, porcine TF as used herein refers to its ECD, e.g., aa to aa 216 of SEQ ID NO: 178. TF is a cell surface receptor for the serine protease Factor VIIa. It is often constitutively expressed in specific cells surrounding blood vessels and in some disease settings.

[0172] In some embodiments, the term TF as used herein refers to a TF epitope.

[0173] The terms "antibody," "immunoglobulin," or "Ig" are used interchangeably herein and are used in the broadest sense to specifically encompass, for example, polyclonal antibodies, monoclonal antibodies (including agonists, antagonists, neutralizing antibodies, and full-length monoclonal antibodies), antibody compositions with polyepitopic or monoepitopic specificity, recombinantly produced antibodies, single domain antibodies, monospecific antibodies, multispecific antibodies (including bispecific 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 TF-binding properties, as well as antibody fragments (and / or polypeptides comprising antibody fragments). Non-limiting examples of antibody fragments include antigen-binding and / or effector regions of antibodies, such as Fab, Fab', F(ab')2, Fv, scFv, (scFv)2, single-chain antibody molecules, dual variable domain antibodies, single variable domain antibodies, linear antibodies, V regions, multispecific antibodies formed from antibody fragments, F(ab)2, Fd, Fc, diabodies, didiabodies, disulfide-linked Fvs (dsFv), single-domain antibodies (e.g., nanobodies), or other fragments (e.g., fragments consisting of non-covalently linked heavy and light chain variable regions). Generally speaking, the variable (V) region can be any suitable arrangement of immunoglobulin heavy (VH) and / or light (VL) chain variable regions. For example, antibodies also include tetrameric antibodies comprising two heavy chain molecules and two light chain molecules, antibody light chain monomers, and antibody heavy chain monomers. Thus, for example, the V region may be dimeric and contain VH-VH, VH-VL, or VL-VL dimers that bind to TF. In either embodiment, the VH and VL regions can be covalently linked, either directly or via a linker, to form a single-chain Fv (scFv). For ease of reference, scFv proteins are referred to herein as being within the category of "antibody fragments." Another form of antibody fragment is a peptide comprising one or more complementarity-determining regions (CDRs) of an antibody.CDRs (also called "minimal recognition units" or "hypervariable regions") can be obtained by constructing a polynucleotide encoding one or more CDRs of interest. Such polynucleotides can be prepared, for example, by using the polymerase chain reaction to synthesize variable regions 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, variable regions of novel antigen receptors (v-NARs), and bis-single-chain Fv regions (see, e.g., Hollinger and Hudson, Nature Biotechnology, 23(9):1126-1136, 2005). In some embodiments, antibodies comprising a VH and / or a 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, antibodies can comprise epitope-binding fragments of any of the above.The antibodies described herein can be of any class (e.g., IgG, IgE, IgM, IgD, and IgA) or any subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2) of immunoglobulin molecule.

[0174] The term "monospecific" as used herein describes an antibody that has one or more binding sites, each of which binds to the same epitope on the same antigen.

[0175] The term "bispecific" means that an antibody can specifically bind to at least two different antigenic determinants; for example, two binding sites are formed by pairing an antibody heavy chain variable region (VH) and an antibody light chain variable region (VL), each binding to a different antigen or a different epitope on the same antigen. Such bispecific antibodies can 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 are the 2+1 or 1+2 formats (containing two binding sites for a first antigen or epitope and one binding site for a second antigen or epitope), or the 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 can bind to a different antigenic determinant. Such bispecific antibodies may bind to two different epitopes on the same antigen (eg, epitopes on TF).

[0176] The term "identical" or percent "identity" in the context of two or more nucleic acids or polypeptides refers to two or more sequences or subsequences that are the same or have a specified percentage of the same nucleotides or amino acid residues when compared and aligned (introducing gaps, if necessary) to obtain maximum correspondence, without considering any conservative amino acid substitutions as part of the sequence identity. Percent identity can be measured using sequence comparison software or algorithms or by visual inspection. Various algorithms and software that can be used to obtain alignments of amino acid or nucleotide sequences are well known in the art. These include, but are not limited to, Basic Local Alignment Search Tool (BLAST), ALIGN, MegAlign, BestFit, GCG Wisconsin Package, and variations thereof. In some embodiments, two nucleic acids or polypeptides are substantially identical when compared and aligned for maximum correspondence, as determined using a sequence comparison algorithm or by visual inspection, meaning 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, identity exists over a region of the amino acid sequence that is at least about 10 residues in length, at least about 20 residues in length, at least about 40-60 residues in length, at least about 60-80 residues in length, or any integer value therebetween. In some embodiments, 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 regions of target proteins or antibodies). In some embodiments, the identity exists over a region of the nucleotide sequence that is at least about 10 bases in length, at least about 20 bases in length, at least about 40-60 bases in length, at least about 60-80 bases in length, or any integer value therebetween.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 sequences are substantially identical over the entire length of the sequences being compared (e.g., nucleotide sequences encoding proteins of interest).

[0177] "Conservative amino acid substitution" refers to a substitution in which one amino acid residue is replaced with 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), uncharged polar 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 phenylalanine for tyrosine is a conservative substitution. In general, conservative substitutions in the sequences of the polypeptides, soluble proteins, and / or antibodies of the present disclosure do not eliminate binding to the target binding site of the polypeptide, soluble protein, or antibody containing the amino acid sequence. Methods for identifying conservative amino acid substitutions that do not eliminate binding are well known in the art.

[0178] The term "polypeptide" refers to a polymer of amino acids of any length. The polymer can be linear or branched, and it can contain modified amino acids and non-amino acids (e.g., non-amino acids can be substituted). The term also encompasses amino acid polymers that are modified naturally or by intervention, such as disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification (e.g., linkage or conjugation (directly or indirectly) with a moiety, such as a labeling component or a drug (e.g., a toxin)). For example, polypeptides containing one or more analogs of an amino acid (e.g., including unnatural amino acids), as well as other modifications known in the art, are also included within the definition. Because the polypeptides of the present disclosure may be based on antibodies or other members of the immunoglobulin superfamily, it is understood that in some embodiments, the polypeptides can exist as single chains or single-chain dimers.

[0179] As used herein, an "antigen" is a moiety or molecule that contains an epitope to which an antibody can bind. Thus, an antigen is capable of being bound by an antibody. In some embodiments, the antigen to which the antibodies described herein bind is TF (e.g., human TF), or a fragment thereof (e.g., a fragment comprising one or more regions of TF).

[0180] As used herein, "epitope" is a term used in the art to refer to a localized region of an antigen to which an antibody can bind. An epitope can be a linear epitope, or a conformational, nonlinear, or discontinuous epitope. For example, in the case of a polypeptide antigen, an epitope can be consecutive amino acids of the polypeptide (a "linear" epitope), or it can include amino acids from two or more noncontiguous regions of the polypeptide (a "conformational," "nonlinear," or "discontinuous" epitope) (e.g., human TF). Those skilled in the art will understand that, in general, a linear epitope 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 they are folded into a native three-dimensional protein structure. In other embodiments, an antibody requires that the amino acid residues comprising the epitope exhibit a particular conformation (e.g., a bend, twist, turn, or fold) in order to recognize and bind to the epitope.

[0181] An antibody binds to an "epitope," "essentially the same 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 competitive assay, which can be configured in a variety of formats (e.g., using either labeled antigen or labeled antibody). In some assays, the antigen is immobilized on a 96-well plate or expressed on a cell surface, and the ability of an unlabeled antibody to block the binding of a labeled antibody is measured using radioactive, fluorescent, or enzymatic labels.

[0182] "Epitope binning" is the process of grouping antibodies based on the epitopes they recognize. More specifically, epitope binning includes methods and systems for clustering antibodies based on their epitope recognition characteristics and distinguishing the epitope recognition characteristics of different antibodies using competitive assays combined with computational processes to identify antibodies with different binding specificities. Additional details regarding epitope binning and methods for determining antibody epitope binding are described herein, as shown in Example 5.

[0183] As used herein, the terms "specifically bind," "specifically recognize," "immunospecifically bind," "selectively bind," "immunospecifically recognize," and "immunospecific" are analogous terms in the context of antibodies and refer to a molecule that binds to an antigen (e.g., an epitope), as would be understood by one of skill in the art.

[0184] In some embodiments, "specifically binds" means, for example, that a polypeptide or molecule interacts with an epitope, protein, or target molecule more frequently, more rapidly, for longer, with higher affinity, or a combination of the above, than with alternative substances (including related and unrelated proteins). For example, a molecule that specifically binds to an antigen may generally bind other peptides or polypeptides with lower affinity as determined by, for example, immunoassays, BIACORE™, KinExA 3000 instruments (Sapidyne Instruments, Boise, ID), OctetQK384 systems (ForteBio, Menlo Park, CA), or other assays known in the art. In some embodiments, an antibody or antigen-binding region binds or specifically binds to an antigen with higher affinity than any cross-reactive antigens as determined using experimental techniques (e.g., radioimmunoassays (RIA) and enzyme-linked immunosorbent assays (ELISA)). Typically, a specific or selective reaction will be at least twice the background signal or noise and may be more than 10 times background. For a discussion of binding specificity, see, e.g., Fundamental Immunology 332-36 (Paul ed., 2d ed. 1989). In some embodiments, the extent of binding of an antibody or antigen-binding region to a "non-target" protein 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 exhibits a K A At least 2 logs, 2.5 logs, 3 logs, 4 logs, or more than K A In some embodiments, a molecule that specifically binds to an antigen does not cross-react with other proteins. In another specific embodiment, a molecule that specifically binds to an antigen does not cross-react with other non-TF proteins. In some embodiments, "specifically binds" refers to, for example, a polypeptide or molecule that binds to a protein or target with a K of about 0.1 mM or less (more typically, less than about 1 μM). DIn 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 Specific binding means binding at a specific site. Due to sequence identity between homologous proteins of different species, specific binding may include polypeptides or molecules that recognize proteins or targets from multiple species. Similarly, due to homology within specific regions of the polypeptide sequences of different proteins, specific binding may include polypeptides or molecules that recognize multiple proteins or targets. It is understood 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 require (but can include) exclusive binding, e.g., binding to a single target. Thus, in some embodiments, a polypeptide or molecule may 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 may contain two identical antigen-binding sites, each of which specifically binds to the same epitope on two or more proteins. In another embodiment, an antibody may be bispecific and contain at least two antigen-binding sites with different specificities. Generally, but not necessarily, reference to "binding" means "specific binding."

[0185] The term "binding affinity" generally refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen such as TF). 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 a binding molecule X for its binding partner Y is generally determined by the dissociation constant (K D) Affinity can be measured by common methods known in the art, including those described herein. Low affinity antibodies generally bind antigens slowly and tend to dissociate easily, whereas high affinity antibodies generally bind antigens quickly and tend to remain bound longer. Various methods for measuring binding affinity are known in the art, any of which can be used for purposes of the present disclosure. In one embodiment, "K" is used. D " or "K D The "K value" can be measured by biolayer interferometry (BLI), for example, using an OctetQK384 system (ForteBio, Menlo Park, CA). Alternatively, the K D may be measured, for example, by a radiolabeled antigen binding assay (RIA) (Chen et al., (1999) J. Mol Biol 293:865-881) performed using a Fab version of the antibody of interest and its antigen, or by using a surface plasmon resonance (SPR) assay by BIACORE™, for example, using a BIACORE™-2000 or BIACORE™-3000 (BIAcore, Inc., Piscataway, NJ). "On rate" or "rate of association" or "association rate" or "k on " and "off rate" or "rate of dissociation" or "dissociation rate" or "k off " can also be determined using the same SPR or BLI techniques described herein, for example, using an OctetQK384 system (ForteBio, Menlo Park, CA) or a BIACORE™-2000 or BIACORE™-3000 (BIACORE™, Inc., Piscataway, NJ), respectively.

[0186] The term "competition," when used in the context of TF antibodies, describes a binding substance whose binding to an epitope or binding site is at least partially inhibited in the presence of another binding substance by the binding of the other binding substance. Competition can be determined in assays in which the binding substance under study blocks or inhibits 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., TF). Various types of competitive binding assays can be used to determine whether a test binding substance competes with a reference molecule for binding to TF (e.g., human TF). Examples of assays that can be used include solid-phase direct or indirect radioimmunoassays (RIA), solid-phase direct or indirect enzyme immunoassays (EIA), sandwich competition assays (see, e.g., Stahl et al., (1983) Methods in Enzymology 9:242-253); solid-phase direct biotin-avidin EIA (see, e.g., Kirkland et al., (1986) J. Immunol. 137:3614-3619 or Cheung et al., (1990) Virology 176:546-552); solid-phase direct label assays, solid-phase direct label sandwich assays (see, e.g., Harlow and Lane, (1988) Antibodies, A Laboratory Manual, Cold Spring Harbor Press); solid-phase direct label RIA using I-125 labels (see, e.g., Morel et al., (1988) Antibodies, A Laboratory Manual, Cold Spring Harbor Press); al., (1988) Molec. Immunol. 25:7-15; and direct labeling RIA (Moldenhauer et al., (1990) Scand. (J. Immunol. 32:77-82). Typically, such assays involve the use of purified antigen (e.g., TF (e.g., human TF)) bound to a solid surface or cells with either an unlabeled test antigen-binding protein (e.g., a test TF antibody or ADC) or a labeled reference antigen-binding protein (e.g., a reference TF antibody or ADC). Competitive inhibition can be measured by determining the amount of label bound to the solid surface or cells in the presence of the test antigen-binding protein.Typically, the test antigen-binding protein is present in excess. Antibodies identified in a competition assay (competing antibodies) include antibodies that bind to the same epitope as the reference antibody and / or antibodies that bind to adjacent epitopes sufficiently close to the epitope bound by the reference antibody (e.g., similar or overlapping epitopes) that steric hindrance of the antibodies occurs. Typically, when present in excess, a competing antibody will inhibit specific binding of the reference antibody to a common antigen 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%, 99%, or more.

[0187] As used herein, the terms "constant region" and "constant domain" are used interchangeably and are well-known antibody terminology, referring to portions of an antibody, e.g., the carboxyl-terminal portions of the light and / or heavy chains, that are not directly involved in binding the antibody to an antigen, but which may exhibit various effector functions (e.g., interaction with Fc receptors). The terms include portions of immunoglobulin molecules that generally have more conserved amino acid sequences than immunoglobulin variable regions.

[0188] Antibody "effector functions" refer to the biological activities attributable to the Fc region of an antibody (e.g., a native sequence Fc region or an amino acid sequence variant Fc region), which vary 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; down-regulation of cell surface receptors (e.g., B cell receptors); and B cell activation.

[0189] The term "Fc region" is used herein to define the C-terminal region of an immunoglobulin heavy chain (including, for example, native sequence Fc regions, recombinant Fc regions, and variant Fc regions). Although the boundaries of the Fc region of an immunoglobulin heavy chain can vary, the human IgG heavy chain Fc region is often defined to stretch from the amino acid residue at position Cys226 (EU numbering system), or from position 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 recombinantly engineering the nucleic acid encoding the antibody heavy chain. Exemplary Fc region sequences are provided below (CH2 domain = bold text, CH3 domain = underlined text): [ka]

[0190] A "functional Fc region" possesses an "effector function" 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; down-regulation of cell surface receptors (e.g., B cell receptor, BCR), and the like. Such effector functions typically require that the Fc region be combined with a binding region or domain (e.g., an antibody variable region or domain), and can be assessed using the various assays disclosed.

[0191] A "native sequence Fc region" comprises an amino acid sequence identical to that of an Fc region found in nature, and has not been manipulated, modified, and / or altered (e.g., isolated, purified, selected, included, or combined with other sequences (e.g., variable region sequences)) by man. Native sequence human Fc regions include native sequence human IgG1 Fc regions (non-A and A allotypes); native sequence human IgG2 Fc regions; native sequence human IgG3 Fc regions; and native sequence human IgG4 Fc regions; and naturally occurring variants thereof.

[0192] A "variant Fc region" comprises an amino acid sequence that differs from that of a native-sequence Fc region by at least one amino acid modification (e.g., substitution, addition, or deletion), preferably one or more amino acid substitution(s). In some embodiments, a variant Fc region comprises at least one amino acid substitution compared to a native-sequence Fc region or the Fc region of a parent polypeptide, e.g., about one to about ten amino acid substitutions, preferably about one to about five amino acid substitutions, in the native-sequence Fc region or the Fc region of a parent polypeptide. The variant Fc regions described herein may have at least about 80% homology to a native-sequence Fc region and / or the Fc region of a parent polypeptide, or at least about 90% homology thereto (e.g., at least about 95% homology). The variant Fc regions described herein may have loss of effector function (e.g., silent Fc).

[0193] The term "heavy chain" when used with respect to an antibody refers to a polypeptide chain of about 50 to 70 kDa, the amino-terminal portion of which contains a variable region of about 120 to 130 or more amino acids, and the carboxy-terminal portion of which contains one or more constant regions. "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., IgG subclasses, e.g., IgG1, IgG2, IgG3, and IgG4), respectively.

[0194] As used herein, the term "light chain" when used in reference to an antibody refers to a polypeptide chain of approximately 25 kDa, the amino-terminal portion of which contains a variable region of about 100 to about 110 or more amino acids, and the carboxy-terminal portion of which contains a constant region. The approximate length of a light chain is 211 to 217 amino acids. There are two different types, e.g., kappa (κ) or lambda (λ), based on the amino acid sequence of the constant region. Light chain amino acid sequences are well known in the art.

[0195] The terms "antigen-binding fragment," "antigen-binding domain," and "antigen-binding region" refer to a portion of an antibody that contains amino acid residues that interact with an antigen and confer specificity and affinity to the binding fragment or region for the antigen (e.g., CDRs). As used herein, "antigen-binding fragment" includes "antibody fragments" that contain a portion of an antibody (e.g., one or more CDRs, e.g., the antigen-binding region or variable region of an antibody).

[0196] Antibodies as described herein include, but are not limited to, synthetic antibodies, monoclonal antibodies, recombinant antibodies, multispecific antibodies (including, e.g., bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, intrabodies, single-chain Fvs (scFvs) (including, e.g., monospecific, bispecific, etc.), camelized antibodies, Fab fragments, F(ab') fragments, disulfide-linked Fvs (sdFvs), anti-idiotypic (anti-Id) antibodies, and epitope-binding fragments of any of the above.

[0197] In some embodiments, the antibodies described herein include immunoglobulin molecules and immunologically active portions of immunoglobulin molecules (e.g., molecules that contain one or more antigen-binding sites that bind to TF).

[0198] The TF antibodies described herein can be of any type (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) of immunoglobulin molecule. In some embodiments, the TF antibodies described herein are IgG antibodies (e.g., human IgG), or classes (e.g., human IgG1, IgG2, IgG3, or IgG4) or subclasses thereof.

[0199] In some embodiments, the TF antibody is a four-chain antibody unit comprising two heavy (H) / light (L) chain pairs, wherein the amino acid sequences of the H chains are identical and the amino acid sequences of the L chains are identical. In some embodiments, the H chain and / or L chain comprise a constant region (e.g., a human constant region). In some embodiments, the L chain constant region of the TF antibody is a κ or λ light chain constant region (e.g., a human κ or λ light chain constant region). In some embodiments, the H chain constant region of the TF antibody comprises a gamma heavy chain constant region (e.g., a human gamma heavy chain constant region). In some embodiments, the TF antibody comprises an IgG constant region, e.g., a human IgG constant region (e.g., an IgG1, IgG2, IgG3, and / or IgG4 constant region).

[0200] As used herein, "TF antibody" and "antibody that binds to TF" are used interchangeably and refer to an antibody that preferentially binds to TF. An antibody or fragment thereof may preferentially bind to TF (e.g., human TF), meaning that the antibody or fragment thereof binds to TF (e.g., human TF) with higher affinity than it binds to an unrelated control protein. For example, an antibody or fragment thereof may specifically recognize and bind to TF or a portion thereof. "Specific binding" means that a TF antibody or fragment thereof binds to TF with an affinity that is at least 5, 10, 15, 20, 25, 50, 100, 250, 500, 1000, or 10,000 times higher than its affinity for an unrelated control protein (e.g., hen egg white lysozyme). In some embodiments, a TF antibody or fragment thereof may bind substantially exclusively to TF (e.g., a measurable difference in binding affinity allows TF to be distinguished from other known polypeptides). In some embodiments, a TF antibody may react with a TF sequence other than a human TF sequence (e.g., a cynomolgus monkey TF sequence).

[0201] The terms "variable region" and "variable domain" are used interchangeably to refer to portions of an antibody's light and heavy chains, generally located at the amino termini of the light and heavy chains, approximately 120-130 amino acids in length for heavy chains and approximately 100-110 amino acids in length for light chains, which are used to determine the binding and specificity of each antibody 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 certain segments of the variable region vary widely among antibodies. The V region mediates antigen binding and defines the specificity of a particular antibody for its antigen. However, variability is not evenly distributed across the 110-amino acid span of the variable region. Instead, the V region consists of stretches of approximately 15-30 amino acids of low variability (e.g., relatively invariant) called framework regions (FRs), separated by shorter regions of high variability (e.g., extreme variability) called "hypervariable regions" or "complementarity-determining regions." The heavy and light chain variable regions each contain four framework regions (FR1, FR2, FR3, and FR4) and primarily adopt a β-sheet configuration connected by three hypervariable regions, which form loops to connect, and in some cases, form part of, the β-sheet structure. The hypervariable regions of each chain are held together in close proximity by the framework regions and, together with the hypervariable regions of the other chain, participate in the formation of the antigen-binding site of antibodies (see, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, (1991)). The constant regions are not directly involved in binding the antibody to the antigen, but exert various effector functions (e.g., antibody participation in antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC)). The sequences of the variable regions vary significantly among different antibodies. The sequence variability is concentrated in the CDRs, while the less variable portions within the variable regions are called framework regions (FRs). The CDRs of the light and heavy chains are primarily responsible for the interaction of the antibody with the antigen.In certain embodiments, the variable region is a human variable region.

[0202] As used herein, the terms "hypervariable region," "HVR," "HV," "complementarity-determining region," and "CDR" refer to regions of an antibody variable region that are hypervariable in sequence and / or form structurally defined loops. Generally, antibodies contain six hypervariable regions: three in the VH (H1 or VH CDR1, H2 or VH CDR2, and H3 or VH CDR3) and three in the VL (L1 or VL CDR1, L2 or VL CDR2, and L3 or VL CDR3). Several hypervariable region delineations are in use and are encompassed herein. Kabat CDRs are based on sequence diversity and are the most widely used (see, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)). Chothia, instead, refers to the location of the structural loops (e.g., Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). The ends of the Chothia CDR-H1 loop, when numbered according to the Kabat numbering convention, vary from H32 to H34 depending on the length of the loop (this is because the Kabat numbering scheme places the insertion at H35A and H35B; if neither 35A nor 35B is present, the loop ends at 32; if only 35A is present, the loop ends at 33; and if both 35A and 35B are present, the loop ends at 34). AbM hypervariable regions are identified according to the Kabat numbering convention. They represent a compromise between the CDRs and Chothia structural loops and are used in Oxford Molecular's AbM antibody modeling software (e.g., Martin, in Antibody Engineering, Vol. 2, Chapter 3, Springer Verlag). The "contact" hypervariable regions are based on an analysis of available complex crystal structures. Residues from each of these hypervariable regions or CDRs are shown below.

[0203] 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 specialized for immunoglobulins (IGs), T cell receptors (TRs), and major histocompatibility complexes (MHCs) from humans and other vertebrates. Herein, CDRs are referred to in terms of both amino acid sequence and location within the light or heavy chain. Because the "location" of CDRs within the structure of immunoglobulin variable regions is conserved across species and resides within structures called loops, CDR and framework residues can be readily identified by using a numbering system that aligns variable region sequences with structural features. This information can be used to graft and replace CDR residues from one immunoglobulin species onto an acceptor framework, typically derived from a human antibody. Additional numbering systems (AHon) have been developed by Honegger and Pluckthun, J. Mol. Biol. 309:657-670 (2001). For example, correspondences between numbering systems, including Kabat numbering and the IMGT® unique numbering system, are well known to those skilled in the art (see, e.g., Kabat, supra; Chothia and Lesk, supra; Martin, supra; Lefranc et al., supra), and are also provided below. The various systems known in the art or described herein represent different ways of depicting CDRs, and are often considered equivalent when used to define the same antibody. The exemplary system provided herein combines Kabat and Chothia. [Table 4]

[0204] The hypervariable regions may include "extended hypervariable regions" as follows: 24-36 or 24-34 (L1), 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3) for VL; 26-35 or 26-35A (H1), 50-65 or 49-65 (H2), and 93-102, 94-102, or 95-102 (H3) for VH. As used herein, the terms "hypervariable region," "HVR," "HV," "complementarity-determining region," or "CDR" are used interchangeably.

[0205] The term "vector" refers to a substance used to carry or contain a nucleic acid sequence (e.g., to introduce a nucleic acid sequence into a host cell). Examples of vectors include expression vectors, plasmids, phage vectors, viral vectors, episomes, and artificial chromosomes, which may contain selection sequences or markers that enable stable integration into a host cell chromosome. Furthermore, a vector may contain one or more selection marker genes and appropriate expression control sequences. For example, selection marker genes that may be included provide resistance to antibiotics or toxins, complement auxotrophic deficiencies, or supply critical nutrients not present in the culture medium. Expression control sequences include constitutive and inducible promoters, transcription enhancers, and transcription terminators, all of which are well known in the art. When two or more nucleic acid molecules (e.g., both the heavy and light chains of an antibody, or both the VH and VL regions of an antibody) are to be coexpressed, both nucleic acid molecules can be inserted, for example, into a single expression vector or into separate expression vectors. For single vector expression, the encoding nucleic acids can be operably linked to one common expression control sequence or can be linked to different expression control sequences (e.g., one inducible promoter and one constitutive promoter). Introduction of the nucleic acid molecule into the host cell can be confirmed using methods well known in the art. Such methods include, for example, nucleic acid analysis (e.g., Northern blotting or polymerase chain reaction (PCR) amplification of mRNA), immunoblotting for expression of the gene product, or other suitable analytical methods for testing the expression of the introduced nucleic acid sequence or its corresponding gene product. Those skilled in the art will understand that the nucleic acid molecule will be expressed in an amount sufficient to produce the desired product (e.g., TF antibody), and will further understand that expression levels can be optimized to obtain sufficient expression using methods well known in the art.

[0206] The terms "TF-mediated disease," "TF-mediated disorder," and "TF-mediated condition" are used interchangeably and refer to any disease, disorder, or condition associated with or characterized by TF-expressing cells (e.g., TF-expressing tumor cells). TF-mediated diseases include cancers (including, but not limited to, cancers that express or overexpress TF).

[0207] As used herein in any embodiment, the term "tumor" refers to any neoplastic cell growth or proliferation, whether malignant or benign, and all pre-cancerous and cancerous cells and tissues.

[0208] The terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth.

[0209] The term "ADC" refers to an antibody-drug conjugate, which in the context of the present invention refers to a TF antibody linked to another moiety, including a drug, as described herein.

[0210] As used herein, "drug" refers to a compound having biological activity, for example, a cytotoxic compound (e.g., a cytotoxic small molecule, a cytotoxic synthetic peptide, etc.).

[0211] Examples of drugs include small molecule drugs (e.g., cancer chemotherapy drugs). For example, if the polypeptide is an antibody (or fragment thereof) with specificity for tumor cells, the antibody can be modified to include modified amino acids as described herein, which can then be conjugated to a cancer chemotherapy drug. Cancer chemotherapy drugs include non-peptide (i.e., non-proteinaceous) compounds that reduce the proliferation of cancer cells, including cytotoxic drugs and cytostatic drugs. 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.

[0212] Suitable cancer chemotherapeutic agents include dolastatin and its active analogs and derivatives, and auristatin and its active analogs and derivatives (e.g., monomethylauristatin D (MMAD), monomethylauristatin E (MMAE), monomethylauristatin F (MMAF), etc.). See, e.g., WO 96 / 33212, WO 96 / 14856, and US 6,323,315. For example, dolastatin 10 or auristatin PE may be included in the TF-ADCs of the present disclosure. Suitable cancer chemotherapeutic agents also include maytansinoids and their active analogs and derivatives (see, e.g., EP 1391213, and Liu et al (1996) Proc. Natl. Acad. Sci. USA 93:8618-8623); duocarmycins and their active analogs and derivatives (including, e.g., the synthetic analogs KW-2189 and CB1-TM1); and benzodiazepines and their active analogs and derivatives (e.g., pyrrolobenzodiazepines (PBDs)).

[0213] Drugs that act to reduce cell proliferation are known in the art and are widely used. Such drugs include alkylating agents (e.g., nitrogen mustards, nitrosoureas, ethyleneimine derivatives, alkylsulfonates, and triazenes), including, but not limited to, mechlorethamine, cyclophosphamide (CYNOTAN™), melphalan (L-sarcolysin), carmustine (BCNU), lomustine (CCNU), semustine (methyl-CCNU), streptozocin, chlorozotocin, uracil mustard, chlormethine, ifosfamide, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, dacarbazine, and temozolomide.

[0214] Antimetabolites include folic acid analogs, pyrimidine analogs, purine analogs, and adenosine deaminase inhibitors, including, but 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-dideazatetrahydrofolic acid (DDATHF), leucovorin, fludarabine phosphate, pentostatin, and gemcitabine.

[0215] Suitable natural products and derivatives thereof (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, brequinar; alkaloids (e.g., vincristine, vinblastine, vinorelbine, vindesine, etc.); podophyllotoxins (e.g., etoposide, teniposide, etc.); antibiotics (e.g., an tracycline, daunorubicin hydrochloride (daunomycin, rubidomycin, cerbidine), idarubicin, doxorubicin, epirubicin, and morpholino derivatives, etc.); phenoxyzolidinib cyclopeptides (e.g., dactinomycin); basic glycopeptides (e.g., bleomycin); anthraquinone glycosides (e.g., plicamycin (mithramycin)); anthracenediones (e.g., mitoxantrone); azirinopyrroloindoleziones (e.g., mitomycin); macrocyclic immunosuppressants (e.g., cyclosporine, FK-506 (tacrolimus, prograf), rapamycin, etc.), etc.

[0216] Other antiproliferative cytotoxic agents are navelbene, CPT-11, anastrazole, letrazole, capecitabine, reloxafine, cyclophosphamide, ifosamide, and droloxafine.

[0217] Microtubule-acting agents 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), dolstatin 10 (NSC 376128), maytansine (NSC 153858), rhizoxin (NSC 332598), paclitaxel (Taxol®), Taxol® derivatives, docetaxel (Taxotere®), thiocolchicine (NSC 361792), trityl cysterol, vinblastine sulfate, vincristine sulfate, natural and synthetic epothilones (including but not limited to eoptilone A, epothilone B, discodermolide; estramustine, nocodazole, and the like).

[0218] Hormone modulating agents and steroids (including synthetic analogs) suitable for use include, but are not limited to, corticosteroids (e.g., prednisone, dexamethasone, etc.), estrogens and pregestins (e.g., hydroxyprogesterone caproate, medroxyprogesterone acetate, megestrol acetate, estradiol, clomiphene, tamoxifen, etc.), adrenocortical suppressants (e.g., aminoglutethimide; 17α-ethinylestradiol; diethylstilbestrol; testosterone, fluoxymesterone, dromostanolone propionate, etc.), and the like. Estrogens stimulate proliferation and differentiation, therefore compounds that bind to the estrogen receptor are used to inhibit this activity. Corticosteroids can inhibit T-cell proliferation.

[0219] Other suitable chemotherapeutic agents include metal complexes (e.g., cisplatin (cis-DDP), carboplatin, etc.); ureas (e.g., hydroxyurea), hydrazines (e.g., N-methylhydrazine); epidophyllotoxins; topoisomerase inhibitors, procarbazine; mitoxantrone; leucovorin; tegafur, etc. Other antiproliferative agents of interest include immunosuppressants (e.g., mycophenolic acid, thalidomide, desoxispargualine, azasporine, leflunomide, mizoribine, azaspirane (SKF 105685), gefitinib (IRESSA®, ZD 1839, 4-(3-chloro-4-fluorophenylamino)-7-methoxy-6-(3-(4-morpholinyl)propoxy)quinazoline), etc.

[0220] 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, such as 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 techniques known to those skilled in the art (see, for example, WO 94 / 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 EP 590,267), or can be obtained from various commercial sources (e.g., Sigma Chemical Co., St. Louis, Missouri (T7402 from Taxus brevifolia or T-1912 from Taxus yannanensis)). Paclitaxel should be understood to refer not only to the common chemically available 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).

[0221] The term "taxane" also encompasses 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 No. WO 99 / 18113; the piperazino and other derivatives described in WO 99 / 14209; the taxane derivatives described in WO 99 / 09021, WO 98 / 22451, and U.S. Pat. No. 5,869,680; the 6-thio derivatives described in WO 98 / 28288; the sulfenamide derivatives described in U.S. Pat. No. 5,821,263; and the taxol derivatives described in U.S. Pat. No. 5,415,869. Additionally, prodrugs of paclitaxel are included, including, but not limited to, those described in WO 98 / 58927, WO 98 / 13059, and U.S. Pat. No. 5,824,701.

[0222] Biological response modifiers suitable for use include, but are not limited to, (1) inhibitors of tyrosine kinase (RTK) activity; (2) inhibitors of serine / threonine kinase activity; (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.

[0223] An "effective amount" is generally an amount sufficient to reduce the severity and / or frequency of symptoms, eliminate symptoms and / or underlying causes, prevent the occurrence of symptoms and / or underlying causes, and / or ameliorate or correct damage caused by or associated with a disease, disorder, or condition. In some embodiments, the effective amount is a therapeutically effective amount.

[0224] As used herein, the term "therapeutically effective amount" refers to an amount of an antibody or ADC described herein sufficient to reduce and / or ameliorate the severity and / or duration of a particular disease, disorder, or condition, and / or its associated symptoms. A therapeutically effective amount of an agent, including a therapeutic agent, can be the amount necessary to (i) reduce or ameliorate the advancement or progression of a given disease, disorder, or condition, (ii) reduce or ameliorate the recurrence, progression, or onset of a given disease, disorder, or condition, and / or (iii) improve or enhance the therapeutic efficacy of another therapy (e.g., a therapy other than administration of an antibody or ADC described herein). A "therapeutically effective amount" of a substance / molecule / agent (e.g., a TF antibody or ADC) of the present disclosure can vary based on a number of factors (e.g., the individual's condition, age, sex, and weight, as well as the ability of the substance / molecule / agent) to elicit a desired response in an individual. A therapeutically effective amount encompasses an amount in which any toxic or adverse effects of the substance / molecule / agent are outweighed by the therapeutically beneficial effects. In some embodiments, the term "therapeutically effective amount" refers to an amount of an antibody or other agent (e.g., or drug) effective to "treat" a disease, disorder, or condition in a subject or mammal.

[0225] In some embodiments, the drug is a microtubule-acting drug with antiproliferative activity (e.g., a maytansinoid). In some embodiments, the drug is an antimitotic drug (e.g., an auristatin or an active auristatin analog or derivative thereof). In some embodiments, the drug is a DNA alkylating agent.

[0226] As used herein, the term "pharmaceutically acceptable" means approved by a regulatory agency of the federal or state government, or listed in the United States Pharmacopoeia, the European Pharmacopoeia, or other generally recognized pharmacopoeias for use in animals, particularly humans.

[0227] "Excipients" include carriers, fillers, preservatives, or stabilizers that are nontoxic to cells or mammals exposed thereto at the dosages and concentrations employed, and may be included, for example, to affect stability, bulk the formulation, or impart a therapeutic effect to the active ingredient in the final dosage form (e.g., to facilitate absorption, reduce viscosity, increase solubility). An "excipient" may be an organic or inorganic, natural or synthetic ingredient with which an active ingredient is combined to facilitate use of the active ingredient (e.g., combined with the active ingredient to facilitate administration of the active ingredient to a subject). Examples of excipients include: buffers (e.g., phosphate, citric acid, and other organic acids); antioxidants (e.g., ascorbic acid); low molecular weight (e.g., less than about 10 amino acid residues) polypeptides; proteins (e.g., serum albumin, gelatin, or immunoglobulins); 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 non-ionic surfactants (e.g., TWEEN®, polyethylene glycol (PEG), and PLURONICS®). The term "excipient" can also refer to a diluent, adjuvant (e.g., Freund's adjuvant (complete or incomplete)), excipient, or vehicle used in administering a therapeutic agent. Such excipients can be sterile liquids (e.g., water and oils, such as those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like). Water is an exemplary excipient when the composition (e.g., pharmaceutical composition) is administered intravenously. Saline solutions, aqueous dextrose, and glycerol solutions can also be used as liquid excipients, particularly for injectable solutions.Suitable excipients (e.g., pharmaceutical excipients) include: starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, nonfat dry milk, glycerol, propylene, glycol, water, ethanol, etc. In any embodiment, the composition may also contain minor amounts of wetting agents, emulsifying agents, or pH buffering agents. 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 grades of 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, The Netherlands). For example, a pharmaceutical compound may contain an effective or therapeutically effective amount of TF-ADC, e.g., in isolated or purified form, together with a suitable amount of excipient to provide a form for proper administration to a subject. The formulation should be compatible with the mode of administration.

[0228] Before the present invention is further described, it is to be understood that this invention is not limited to particular embodiments described, as such may, of course, vary. It is also understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, as the scope of the present invention will be limited only by the appended claims.

[0229] When a range of values ​​is provided, it is understood that each intervening value (to the nearest tenth of the lower limit, unless the context clearly indicates otherwise), between the upper and lower limit of that range, and any other stated or intervening value in the stated range, is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and, subject to any specifically excluded limit in the stated range, are also encompassed within the invention. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

[0230] It will be understood that certain features of the invention that are described, for clarity, in the context of separate embodiments may also be provided in combination in a single embodiment. Conversely, various features of the invention that are described, for brevity, in the context of a single embodiment may also be provided separately or in any suitable combination. All combinations of embodiments related to the present invention are specifically embraced by the present invention and are disclosed herein as if each and every combination were individually and explicitly disclosed, so long as such combinations include subject matter that is, for example, a stable compound (i.e., a compound that can be produced, isolated, characterized, and tested for biological activity). In addition, all subcombinations of various embodiments and elements thereof (e.g., elements of chemical groups listed in the embodiments describing such variables) are also specifically embraced by the present invention and are disclosed herein as if each and every such subcombination were individually and explicitly disclosed herein.

[0231] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, the preferred methods and materials are described herein. All publications mentioned herein are incorporated by reference to disclose and describe the methods and / or materials in which the publications are cited.

[0232] It should be noted that, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It is further noted that the claims may be drafted to exclude any element. Accordingly, this statement is intended to serve as a predicate for using such exclusive terminology, such as "only," "only," etc., in connection with the recitation of claim elements, or for using a "negative" limitation.

[0233] It will be appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable combination.

[0234] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein should be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the publication dates provided may be different from the actual publication dates, which may need to be independently confirmed.

[0235] TF-DAC An antibody that binds to TF (also referred to herein as a "TF antibody," "anti-TF antibody," "TFAb," "Ab," or "antibody") and a drug can be directly or indirectly linked to each other via a pyridazine-pyrrolo coupling moiety to form a TF-ADC as described herein. In certain embodiments, the TF antibody and two or more drugs or active agents can be linked to each other via one or more functional groups and covalent bonds. For example, the one or more functional groups and covalent bonds can include a branched linker as described herein.

[0236] A moiety of interest (e.g., a drug or active agent) can be conjugated to a TF antibody at any desired site on the antibody. Thus, for example, the present disclosure provides TF antibodies having 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 position at or near the N-terminus of the antibody, and a position between the C-terminus and N-terminus of the antibody (e.g., an internal site on the antibody). Combinations of the above conjugation sites are also possible.

[0237] In certain embodiments, the conjugates of the present disclosure comprise two (or more) drugs or active agents conjugated to the alpha carbons of amino acid residues of a TF antibody. In other words, the conjugates comprise a TF antibody in which the side chains of amino acid residues in the antibody have been modified and are attached to two (or more) drugs or active agents (e.g., attached to the two drugs or active agents via a branched linker as described herein). For example, the conjugates comprise a TF antibody in which the alpha carbons of amino acid residues in the antibody have been modified and are attached to two drugs or active agents (e.g., attached to the two drugs or active agents via a branched linker as described herein).

[0238] Embodiments of the present disclosure include conjugates in which a TF antibody is conjugated to two or more moieties (e.g., three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, or twenty or more moieties). These moieties may be conjugated to the TF antibody at multiple sites within the antibody. In some embodiments, two moieties may be conjugated to a single amino acid residue of the TF antibody. For example, two moieties may be conjugated to the same amino acid residue of the TF antibody. In other embodiments, two moieties are conjugated to the first amino acid residue of the TF antibody, and the other two moieties are conjugated to the second amino acid residue of the TF antibody. For example, a TF antibody may be conjugated to the first and second moieties at the first amino acid residue, the third and fourth moieties at the second amino acid residue, etc. In some cases, two or more amino acid residues of the TF antibody are each conjugated to a pair of moieties (i.e., two moieties), and each pair of moieties is conjugated to the TF antibody via a branched linker as described herein. In some cases, one amino acid residue in the TF antibody is conjugated to a pair of moieties 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 the TF antibody are each conjugated to a pair of moieties via a branched linker as described herein.

[0239] One or more amino acid residues of the TF antibody conjugated to the moiety of interest may be a natural amino acid, a non-natural amino acid, or a combination thereof. For example, the conjugate may include a moiety of interest (e.g., a drug or active agent) conjugated to a natural amino acid residue of the TF antibody. In another example, the conjugate may include a moiety of interest conjugated to a non-natural amino acid residue of the TF antibody. The moiety of interest may be conjugated to the TF antibody at a single natural or non-natural amino acid residue as described above. One or more natural or non-natural amino acid residues in the TF antibody may be conjugated to the moiety of interest described herein. For example, two (or more) amino acid residues (e.g., natural or non-natural amino acid residues) in the TF antibody may be conjugated to two moieties via branched linkers, respectively, resulting in multiple sites in the TF antibody being conjugated to the moiety of interest.

[0240] As described herein, a TF antibody can be conjugated to two or more moieties of interest. In certain embodiments, the moiety of interest is a payload, e.g., a chemical entity (e.g., a drug, an active agent, or a detectable label). For example, a drug (or an active agent, e.g., a cytokine) can be conjugated to a TF antibody, or in other embodiments, a detectable label can be conjugated to a TF antibody. In other embodiments, a combination of different payloads can be conjugated to a TF antibody. Thus, for example, embodiments of the present disclosure include, but are not limited to, conjugates of a TF antibody with two or more drugs, conjugates of a TF antibody with two or more active agents (e.g., cytokines), conjugates of a TF antibody with two or more detectable labels, and combinations thereof.

[0241] In certain embodiments, the TF antibody and the moiety of interest (e.g., a drug or active agent) are conjugated via a conjugation moiety. For example, the TF antibody and the moiety of interest can each be bound (e.g., covalently linked) to a conjugation moiety, thereby indirectly linking the TF antibody and the moiety of interest via the conjugation moiety. In some cases, the conjugation moiety comprises a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl compound, or a derivative of a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl compound. A general scheme for attaching a coupling moiety of interest to a TF antibody, for example, via a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety, is shown in the following general reaction scheme: Hydrazinyl-indolyl and hydrazinyl-pyrrolo-pyridinyl conjugation moieties are also referred to herein as hydrazino-iso-Picteth-Spengler (HIPS) conjugation moieties and aza-hydrazino-iso-Picteth-Spengler (azaHIPS) conjugation moieties, respectively. Thus, one of skill in the art will understand that in some embodiments, the carbon shown directly adjacent to Ab in a formula disclosed herein (e.g., Formula (I) or (II)) was originally part of the antibody prior to conjugation. In some embodiments, this carbon is conjugated to a (fGly') residue, thereby conjugating the antibody to the linker payload. In other embodiments, this carbon is interpreted as part of the (fGly') residue that conjugates the antibody to the linker payload.

[0242] [ka]

[0243] In the above reaction scheme, each R independently comprises a moiety of interest (e.g., a drug or active agent) conjugated to a TF antibody (e.g., conjugated to a TF antibody via a linker described herein), and n is an integer from 1 to 4. As shown in the above reaction scheme, a conjugation moiety (e.g., a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety) is attached to two or more drugs or active agents R. A TF antibody containing a 2-formylglycine residue (fGly) reacts with the conjugation moiety to generate a TF antibody conjugate, whereby two or more drugs or active agents are attached to the TF antibody via the conjugation moieties.

[0244] As described herein, a moiety can be any of a variety of moieties (including, but not limited to, a chemical entity such as a detectable label, or a drug or active agent). R' and R" can each independently be any desired substituent (including, 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, alkylamido, substituted alkylamido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl). Z is CR 21 , N.R. 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.

[0245] 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 attached (e.g., covalently attached) to two or more linkers. Thus, embodiments of the present disclosure include hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moieties attached to two or more drugs or active agents via corresponding linkers. Thus, conjugates of the present disclosure may include two or more linkers, each linker attaching a corresponding drug or active agent to the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety. Thus, the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety and two or more linkers may be considered collectively as a "branched linker," in which the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety is attached to two or more "branches," each branch comprising a linker attached to a drug or active agent.

[0246] A combination of the same or different payloads can be conjugated to a TF antibody via a branched linker. In certain embodiments, the two payloads (e.g., drugs, active agents, or detectable labels) attached to the branched linker are the same payload (e.g., drugs, active agents, or detectable labels). For example, the first branch of the branched linker can be attached to a payload (e.g., a drug, active agent, or detectable label), and the second branch of the branched linker can be attached to the same payload (e.g., a drug, active agent, or detectable label) as the first branch.

[0247] In other embodiments, the two payloads (e.g., drugs, active agents, or detectable labels) attached to the branched linker are different payloads (e.g., drugs, active agents, or detectable labels). For example, a first branch of the branched linker may be attached to a first payload (e.g., a first drug, active agent, or detectable label), and a second branch of the branched linker may be attached to a second payload (e.g., a second drug, active agent, or detectable label) that is different from the first payload (e.g., the first drug, active agent, or detectable label) attached to the first branch.

[0248] In some embodiments, when two different drugs or active agents are attached to a branched linker, the drugs or active agents can be selected from drugs and active agents that have a synergistic therapeutic effect. "Synergistic," "synergistic effect," or "synergistic effect" refers to a therapeutic effect that is greater than the sum of the effects of the drugs or active agents taken separately. For example, in some cases, using two different drugs or active agents attached to a branched linker can reduce the therapeutically effective concentration at which both payloads act, increasing the overall efficacy of the ADC.

[0249] In some embodiments, when two different drugs or active agents are attached to a branched linker, the drugs or active agents may be selected from drugs and active agents that provide an enhanced therapeutic effect compared to the drugs or active agents used separately. For example, the drugs or active agents may provide enhanced drug delivery for the ADC (e.g., some payloads (e.g., iRGD peptides) may increase extravasation into tissues and enhance tumor infiltration).

[0250] In some embodiments, when two different drugs or active agents are attached to a branched linker, the drugs or active agents may be selected from drugs and active agents that use different mechanisms of action. In some cases, targeting multiple pathways may provide a reduction in tumor drug resistance. Examples of payload combinations may include, but are not limited to, cytotoxic drugs, immunomodulatory molecules that activate or inhibit immune cell populations, cytokines, hormones, radioisotopes-loaded chelators, etc.

[0251] In some embodiments, two different payloads are attached to the branched linker, and the payloads may be selected from a combination of a drug or active agent and a detectable label. For example, one payload may be a detectable label used as an imaging agent or tracer to detect the location of the ADC in vivo, while the second payload may be a drug or active agent that provides a therapeutic effect.

[0252] Various embodiments of linkers that can connect hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moieties to drugs or active agents are described in detail herein.For example, in some examples, the linker is a cleavable linker (e.g., a cleavable linker) described herein.

[0253] In certain embodiments, a TF antibody can be conjugated to two or more moieties of interest, and one or more amino acids of the TF antibody are modified prior to conjugation to the moieties of interest. Modifying one or more amino acids of the TF antibody can result in a TF antibody containing one or more reactive groups suitable for conjugation to the moieties of interest. In some cases, the TF antibody may contain one or more modified amino acid residues to provide one or more reactive groups suitable for conjugation to the moieties of interest (e.g., two or more sites are attached to conjugation moieties (e.g., hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moieties as described above)). For example, an amino acid of the TF antibody can be modified to contain a reactive aldehyde group (e.g., a reactive aldehyde). The reactive aldehyde 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 to contain 2-formylglycine residues (referred to herein as "fGly") by the action of formylglycine generating enzyme (FGE). The fGly residues generated by FGE are also referred to as "formylglycines." In other words, the term "aldehyde tag" is used herein to refer to a "converted" sulfatase motif (i.e., 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 (i.e., a sulfatase motif in which the cysteine ​​or serine residue has not been converted to fGly by FGE, but can be converted (e.g., an unconverted sulfatase motif having the sequence LCTPSR, SEQ ID NO: 100). "Conversion," as used in the context of the action of formylglycine generating enzyme (FGE) on a sulfatase motif, refers to the biochemical modification of a cysteine ​​or serine residue within the sulfatase motif to a formylglycine (fGly) residue (e.g., Cys to fGly or Ser to fGly).Further aspects of aldehyde tags and their use in site-specific protein modification are described in U.S. Pat. No. 7,985,783 and U.S. Pat. No. 8,729,232, the disclosures of each of which are incorporated herein by reference.

[0254] In some cases, a TF antibody containing an fGly residue can be conjugated to a moiety of interest by reacting the fGly with a compound (e.g., a compound containing a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety, as described above) to produce a conjugate. For example, an fGly-containing TF antibody can be contacted with a reaction partner under conditions suitable to provide for conjugation of two or more drugs to the TF antibody. In some cases, the reaction partner can include a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety, as described above. For example, two or more drugs or active agents can be attached to the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety. In some cases, drugs or active agents are attached to the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety (e.g., covalently attached to the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl), with each drug or active agent being attached to the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety via a corresponding linker.

[0255] In certain embodiments, the conjugates of the present disclosure comprise a TF antibody having at least one amino acid residue conjugated to two or more moieties of interest (e.g., drugs or active agents). To create the conjugate, the amino acid residue of the TF antibody may be modified and then conjugated to two or more drugs or active agents that are conjugated to hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moieties, as described above. In certain embodiments, the amino acid residue of the TF antibody is a cysteine ​​or serine residue that has been modified to an fGly residue, as described above. In certain embodiments, the modified amino acid residue (e.g., an fGly residue) is conjugated to two or more drugs or active agents containing hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moieties, as described above, and the conjugates of the present disclosure are provided in which the two or more drugs or active agents are conjugated to the TF antibody via hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moieties. As used herein, the term "fGly" refers to the amino acid residue of the TF antibody that is attached to a moiety of interest (eg, a drug or active agent).

[0256] In certain embodiments, the conjugate comprises a TF antibody having at least one amino acid residue attached to a branched linker described herein, which in turn is attached to two or more drugs or active agents. For example, the conjugate may comprise a TF antibody having at least one amino acid residue (fGly') conjugated to a moiety of interest (e.g., a drug or active agent) as described above.

[0257] Embodiments of the present disclosure include conjugates of formula (I): [ka] (In the formula, Ab represents an antibody that binds to TF; Z 1 , Z 2 , Z 3 , and Z 4 However, each independently, CR4 , N and CL B -W 2 and at least one Z 1 , Z 2 , Z 3 , and Z 4 But CL B -W 2 and R 1 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; R 2 and R 3 are each 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, alkylamido, substituted alkylamido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; or R 2 and R 3 are optionally annularly joined to form a 5- or 6-membered heterocyclyl; Each R 4 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, alkylamido, substituted alkylamido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; L A is the first linker, L B is the second linker, s is an integer from 1 to 10, W 1 is the first drug, W 2 is the second drug).

[0258] The substituents associated with the conjugates of formula (I) are described in more detail below.

[0259] In certain embodiments, Z 1 , Z 2 , Z 3 , and Z 4 However, each independently, CR 4 , N and CL B -W 2 and at least one Z 1 , Z 2 , Z 3 , and Z 4 But CL B -W 2 In certain embodiments, Z 1 But, CR 4 In certain embodiments, Z 1 is N. In certain embodiments, Z 1 But CL B -W 2 In certain embodiments, Z 2 But, CR 4 In certain embodiments, Z 2 is N. In certain embodiments, Z 2 But CL B -W 2 In certain embodiments, Z 3 But, CR 4 In certain embodiments, Z 3 is N. In certain embodiments, Z 3 But CL B -W 2 In certain embodiments, Z 4 But, CR 4 In certain embodiments, Z 4 is N. In certain embodiments, Z 4 But CL B -W2 In some embodiments, Z 1 , Z 3 , and Z 4 Each of these is CR 4 In some embodiments, Z 3 But CL B -W 2 is.

[0260] Various Z 1 , Z 2 , Z 3 , Z 4 For example, in some cases, Z 1 But CL B -W 2 , Z 2 But, CR 4 and Z 3 But, CR 4 and Z 4 But, CR 4 In some cases, Z 1 But, CR 4 and Z 2 But CL B -W 2 and Z 3 But, CR 4 and Z 4 But, CR 4 In some cases, Z 1 But, CR 4 and Z 2 But, CR 4 and Z 3 But CL B -W 2 and Z 4 But, CR 4 In some cases, Z 1 But, CR 4 and Z 2 But, CR 4 and Z 3 But, CR 4 and Z 4 But CL B -W 2 is.

[0261] In certain embodiments, R1 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. 1 is hydrogen. In certain embodiments, R 1 is alkyl or substituted alkyl (e.g., C 1-6 Alkyl or C 1-6 Substituted alkyl, or C 1-4 Alkyl or C 1-4 Substituted alkyl, or C 1-3 Alkyl or C 1-3 In certain embodiments, R 1 is an alkenyl or substituted alkenyl (e.g., C 2-6 Alkenyl or C 2-6 substituted alkenyl, or C 2-4 Alkenyl or C 2-4 substituted alkenyl, or C 2-3 Alkenyl or C 2-3 In certain embodiments, R 1 is alkynyl or substituted alkynyl (e.g., C 2-6 Alkenyl or C 2-6 substituted alkenyl, or C 2-4 Alkenyl or C 2-4 substituted alkenyl, or C 2-3 Alkenyl or C 2-3 In certain embodiments, R 1 is an aryl or substituted aryl (e.g., C 5-8 Aryl or C 5-8 substituted aryl, e.g., C aryl or C substituted aryl, or C aryl or C substituted aryl). In certain embodiments, R 1 is heteroaryl or substituted heteroaryl (e.g., C 5-8 Heteroaryl or C 5-8In certain embodiments, R is a C heteroaryl or a C substituted heteroaryl, such as a C heteroaryl or a C substituted heteroaryl. 1 is a cycloalkyl or substituted cycloalkyl (e.g., C 3-8 Cycloalkyl or C 3-8 Substituted cycloalkyl, e.g., C 3-6 Cycloalkyl or C 3-6 substituted cycloalkyl, or C 3-5 Cycloalkyl or C 3-5 In certain embodiments, R 1 is heterocyclyl or substituted heterocyclyl (e.g., C 3-8 Heterocyclyl or C 3-8 Substituted heterocyclyl, e.g., C 3-6 Heterocyclyl or C 3-6 substituted heterocyclyl, or C 3-5 Heterocyclyl or C 3-5 substituted heterocyclyl).

[0262] In certain embodiments, R 2 and R 3 are each 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, alkylamido, substituted alkylamido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl; or R 2 and R 3 are optionally cyclically joined to form a 5- or 6-membered heterocyclyl.

[0263] In certain embodiments, R 2is 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, alkylamido, substituted alkylamido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. 2 is hydrogen. In certain embodiments, R 2 is alkyl or substituted alkyl (e.g., C 1-6 Alkyl or C 1-6 Substituted alkyl, or C 1-4 Alkyl or C 1-4 Substituted alkyl, or C 1-3 Alkyl or C 1-3 In certain embodiments, R 2 is methyl. In certain embodiments, R 2 is an alkenyl or substituted alkenyl (e.g., C 2-6 Alkenyl or C 2-6 substituted alkenyl, or C 2-4 Alkenyl or C 2-4 substituted alkenyl, or C 2-3 Alkenyl or C 2-3 In certain embodiments, R 2 is alkynyl or substituted alkynyl. In certain embodiments, R 2 is alkoxy or substituted alkoxy. In certain embodiments, R 2 is amino or substituted amino. In certain embodiments, R 2 is carboxyl or carboxyl ester. In certain embodiments, R 2 is acyl or acyloxy. In certain embodiments, R 2 is acylamino or aminoacyl. In certain embodiments, R 2 is an alkylamide or a substituted alkylamide. In certain embodiments, R2 is sulfonyl. In certain embodiments, R 2 is thioalkoxy or substituted thioalkoxy. In certain embodiments, R 2 is an aryl or substituted aryl (e.g., C 5-8 Aryl or C 5-8 substituted aryl, e.g., C aryl or C substituted aryl, or C aryl or C substituted aryl). In certain embodiments, R 2 is heteroaryl or substituted heteroaryl (e.g., C 5-8 Heteroaryl or C 5-8 In certain embodiments, R is a C heteroaryl or a C substituted heteroaryl, such as a C heteroaryl or a C substituted heteroaryl. 2 is a cycloalkyl or substituted cycloalkyl (e.g., C 3-8 Cycloalkyl or C 3-8 Substituted cycloalkyl, e.g., C 3-6 Cycloalkyl or C 3-6 substituted cycloalkyl, or C 3-5 Cycloalkyl or C 3-5 In certain embodiments, R 2 is heterocyclyl or substituted heterocyclyl (e.g., C 3-6 Heterocyclyl or C 3-6 substituted heterocyclyl, or C 3-5 Heterocyclyl or C 3-5 substituted heterocyclyl).

[0264] In certain embodiments, R 3is 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, alkylamido, substituted alkylamido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. 3 is hydrogen. In certain embodiments, R 3 is alkyl or substituted alkyl (e.g., C 1-6 Alkyl or C 1-6 Substituted alkyl, or C 1-4 Alkyl or C 1-4 Substituted alkyl, or C 1-3 Alkyl or C 1-3 In certain embodiments, R 3 is methyl. In certain embodiments, R 3 is an alkenyl or substituted alkenyl (e.g., C 2-6 Alkenyl or C 2-6 substituted alkenyl, or C 2-4 Alkenyl or C 2-4 substituted alkenyl, or C 2-3 Alkenyl or C 2-3 In certain embodiments, R 3 is alkynyl or substituted alkynyl. In certain embodiments, R 3 is alkoxy or substituted alkoxy. In certain embodiments, R 3 is amino or substituted amino. In certain embodiments, R 3 is carboxyl or carboxyl ester. In certain embodiments, R 3 is acyl or acyloxy. In certain embodiments, R 3 is acylamino or aminoacyl. In certain embodiments, R 3 is an alkylamide or a substituted alkylamide. In certain embodiments, R3 is sulfonyl. In certain embodiments, R 3 is thioalkoxy or substituted thioalkoxy. In certain embodiments, R 3 is an aryl or substituted aryl (e.g., C 5-8 Aryl or C 5-8 substituted aryl, e.g., C aryl or C substituted aryl, or C aryl or C substituted aryl). In certain embodiments, R 3 is heteroaryl or substituted heteroaryl (e.g., C 5-8 Heteroaryl or C 5-8 In certain embodiments, R is a C heteroaryl or a C substituted heteroaryl, such as a C heteroaryl or a C substituted heteroaryl. 3 is a cycloalkyl or substituted cycloalkyl (e.g., C 3-8 Cycloalkyl or C 3-8 Substituted cycloalkyl, e.g., C 3-6 Cycloalkyl or C 3-6 substituted cycloalkyl, or C 3-5 Cycloalkyl or C 3-5 In certain embodiments, R 3 is heterocyclyl or substituted heterocyclyl (e.g., C 3-8 Heterocyclyl or C 3-8 Substituted heterocyclyl, e.g., C 3-6 Heterocyclyl or C 3-6 substituted heterocyclyl, or C 3-5 Heterocyclyl or C 3-5 substituted heterocyclyl).

[0265] In certain embodiments, R 2 and R 3 are both methyl.

[0266] In certain embodiments, R 2 and R 3 are optionally cyclically linked to form a 5- or 6-membered heterocyclyl. In certain embodiments, R2 and R 3 are cyclically linked to form a 5- or 6-membered heterocyclyl. In certain embodiments, R 2 and R 3 are cyclically linked to form a 5-membered heterocyclyl. In certain embodiments, R 2 and R 3 is cyclically linked to form a 6-membered heterocyclyl.

[0267] In certain embodiments, each R 4 is 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, alkylamido, substituted alkylamido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

[0268] Each R 4 The various possibilities for R are detailed below. In certain embodiments, R 4 is hydrogen. In certain embodiments, each R 4 is hydrogen. In certain embodiments, R 4 is halogen (e.g., F, Cl, Br, or I). In certain embodiments, R 4 is F. In certain embodiments, R 4 is Cl. In certain embodiments, R 4 is Br. In certain embodiments, R 4 is I. In certain embodiments, R 4 is alkyl or substituted alkyl (e.g., C 1-6 Alkyl or C 1-6 Substituted alkyl, or C 1-4 Alkyl or C 1-4 Substituted alkyl, or C 1-3 Alkyl or C 1-3 In certain embodiments, R4 is methyl. In certain embodiments, R 4 is an alkenyl or substituted alkenyl (e.g., C 2-6 Alkenyl or C 2-6 substituted alkenyl, or C 2-4 Alkenyl or C 2-4 substituted alkenyl, or C 2-3 Alkenyl or C 2-3 In certain embodiments, R 4 is alkynyl or substituted alkynyl. In certain embodiments, R 4 is alkoxy or substituted alkoxy. In certain embodiments, R 4 is amino or substituted amino. In certain embodiments, R 4 is carboxyl or carboxyl ester. In certain embodiments, R 4 is 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 sulfonyl. In certain embodiments, R 4 is thioalkoxy or substituted thioalkoxy. In certain embodiments, R 4 is an aryl or substituted aryl (e.g., C 5-8 Aryl or C 5-8 substituted aryl, e.g., C aryl or C substituted aryl, or C aryl or C substituted aryl (e.g., phenyl or substituted phenyl). In certain embodiments, R 4 is heteroaryl or substituted heteroaryl (e.g., C 5-8 Heteroaryl or C 5-8 In certain embodiments, R is a C heteroaryl or a C substituted heteroaryl, such as a C heteroaryl or a C substituted heteroaryl. 4 is a cycloalkyl or substituted cycloalkyl (e.g., C 3-8 Cycloalkyl or C3-8 Substituted cycloalkyl, e.g., C 3-6 Cycloalkyl or C 3-6 substituted cycloalkyl, or C 3-5 Cycloalkyl or C 3-5 In certain embodiments, R 4 is heterocyclyl or substituted heterocyclyl (e.g., C 3-8 Heterocyclyl or C 3-8 Substituted heterocyclyl, e.g., C 3-6 Heterocyclyl or C 3-6 substituted heterocyclyl, or C 3-5 Heterocyclyl or C 3-5 substituted heterocyclyl).

[0269] In certain embodiments, L A is the first linker. Examples of linkers that can be used in the conjugates of the present disclosure are detailed below.

[0270] In certain embodiments, L B is the second linker. Examples of linkers that can be used in the conjugates of the present disclosure are detailed below.

[0271] In certain embodiments, W 1 is the first drug (or first active agent). Examples of drugs and active agents that can be used in the conjugates of the present disclosure are detailed below.

[0272] In certain embodiments, W 2 is a second drug (or second active agent). Examples of drugs and active agents that can be used in the conjugates of the present disclosure are detailed below.

[0273] In certain embodiments, Ab refers to an antibody that binds to tissue factor ("TF antibody"). In certain embodiments, Ab comprises one or more fGly' residues as described herein. In certain embodiments, the TF antibody is attached to the remainder of the conjugate via an fGly' residue, as described herein. Examples of TF antibodies that can be used in the conjugates of the present disclosure are detailed below.

[0274] In certain embodiments, the conjugate of Formula (I) comprises a first linker, L A The first linker, L A may be utilized to attach a first moiety of interest (e.g., a first drug or active agent) to a TF antibody via a conjugation moiety. A can be attached (e.g., covalently attached) to a conjugation moiety (e.g., as described herein). For example, a first linker, L A can attach a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety to a first drug. The hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety can be attached to a first linker, L A (and thus the first drug) can be used to conjugate to the TF antibody.

[0275] For example, as shown in formula (I) above, L A is attached to the Ab via a conjugation moiety, such that the Ab is linked to the linker L via a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety. A As mentioned above, the Ab is a TF antibody, thereby binding to L A is attached to the TF antibody via a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety, e.g., a linker L A is indirectly attached to the TF antibody via a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety.

[0276] The first linker L in the subject conjugates and compounds A Any convenient linker may be utilized. In certain embodiments, the first linker L A may comprise a group selected from alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acylamino, alkylamido, substituted alkylamido, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, the first linker L A may comprise an alkyl or substituted alkyl group. In certain embodiments, the first linker L A may comprise an alkenyl or substituted alkenyl group. In certain embodiments, the first linker L A may comprise an alkynyl or substituted alkynyl group. In certain embodiments, the first linker, L A may comprise an alkoxy or substituted alkoxy group. In certain embodiments, the first linker L A may comprise an amino or substituted amino group. In certain embodiments, the first linker L A may contain a carboxyl or carboxyl ester group. In certain embodiments, the first linker L A may comprise an acylamino group. In certain embodiments, the first linker L A may comprise an alkylamide or substituted alkylamide group. In certain embodiments, the first linker, L A may comprise an aryl or substituted aryl group. In certain embodiments, the first linker L A may comprise a heteroaryl or substituted heteroaryl group. In certain embodiments, the first linker, L A may comprise a cycloalkyl or substituted cycloalkyl group. In certain embodiments, the first linker, L Amay contain a heterocyclyl or substituted heterocyclyl group.

[0277] In certain embodiments, the first linker L A may comprise 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, copolymers of ethylene glycol and propylene glycol (e.g., the homopolymers and copolymers are unsubstituted or substituted at one end with an alkyl group), polyvinyl alcohol, polyvinyl ethyl ether, polyvinylpyrrolidone, combinations thereof, and the like. In certain embodiments, the polymer is a polyalkylene glycol. In certain embodiments, the polymer is polyethylene glycol. Other linkers can be used, as indicated in the conjugates and compounds described in more detail below.

[0278] In some embodiments, L A is a first linker described by the formula: -(L 1 ) a -(L 2 ) b -(L 3 ) c -(L 4 ) d -(L 5 ) e -(L 6 ) f -, L 1 , L 2 , L 3 , L 4 , L 5 , and L 6 are each independently a linker subunit; and a, b, c, d, e, and f are each independently 0 or 1, with the proviso that at least one of a, b, c, d, e, and f is 1.

[0279] In certain embodiments, the sum of a, b, c, d, e, and f is 1 to 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 each 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.

[0280] In certain embodiments, the linker subunit L 1 is attached to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety (e.g., as shown in formula (I) above). In certain embodiments, the linker subunit L 2 If present, the first drug or active agent W 1 In certain embodiments, the linker subunit L 3 If present, the first drug or active agent W 1 In certain embodiments, the linker subunit L 4 If present, the first drug or active agent W 1 In certain embodiments, the linker subunit L 5 If present, the first drug or active agent W 1 In certain embodiments, the linker subunit L 6 If present, the first drug or active agent W1 is bonded to.

[0281] First linker L A Any convenient linker subunit may be used in the present invention. Linker subunits of interest include, but are not limited to, units of polymers (e.g., polyethylene glycol, polyethylene and polyacrylate, amino acid residue(s), carbohydrate-based 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 (when 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 comprising alkylenediamines).

[0282] In some embodiments, L 1 (when present) comprises polyethylene glycol, modified polyethylene glycol, an amino acid residue, an alkyl group, a substituted alkyl, an aryl group, a substituted aryl group, or a diamine. In some embodiments, L 1 In some embodiments, L 1 In some embodiments, L comprises a modified polyethylene glycol. 1 In some embodiments, L 1 In some embodiments, L 1 comprises an aryl group or a substituted aryl group. In some embodiments, L 1 comprises a diamine (eg, a linking group comprising an alkylenediamine).

[0283] In some embodiments, L 2(when present) comprises polyethylene glycol, modified polyethylene glycol, an amino acid residue, an alkyl group, a substituted alkyl, an aryl group, a substituted aryl group, or a diamine. In some embodiments, L 2 In some embodiments, L 2 In some embodiments, L comprises a modified polyethylene glycol. 2 In some embodiments, L 2 In some embodiments, L 2 comprises an aryl group or a substituted aryl group. In some embodiments, L 2 comprises a diamine (eg, a linking group comprising an alkylenediamine).

[0284] In some embodiments, L 3 (when present) comprises polyethylene glycol, modified polyethylene glycol, an amino acid residue, an alkyl group, a substituted alkyl, an aryl group, a substituted aryl group, or a diamine. In some embodiments, L 3 In some embodiments, L 3 In some embodiments, L comprises a modified polyethylene glycol. 3 In some embodiments, L 3 In some embodiments, L 3 comprises an aryl group or a substituted aryl group. In some embodiments, L 3 comprises a diamine (eg, a linking group comprising an alkylenediamine).

[0285] In some embodiments, L 4 (when present) comprises polyethylene glycol, modified polyethylene glycol, an amino acid residue, an alkyl group, a substituted alkyl, an aryl group, a substituted aryl group, or a diamine. In some embodiments, L 4 In some embodiments, L 4In some embodiments, L comprises a modified polyethylene glycol. 4 In some embodiments, L 4 In some embodiments, L 4 comprises an aryl group or a substituted aryl group. In some embodiments, L 4 comprises a diamine (eg, a linking group comprising an alkylenediamine).

[0286] In some embodiments, L 5 (when present) comprises polyethylene glycol, modified polyethylene glycol, an amino acid residue, an alkyl group, a substituted alkyl, an aryl group, a substituted aryl group, or a diamine. In some embodiments, L 5 In some embodiments, L 5 In some embodiments, L comprises a modified polyethylene glycol. 5 In some embodiments, L 5 In some embodiments, L 5 comprises an aryl group or a substituted aryl group. In some embodiments, L 5 comprises a diamine (eg, a linking group comprising an alkylenediamine).

[0287] In some embodiments, L 6 (when present) comprises polyethylene glycol, modified polyethylene glycol, an amino acid residue, an alkyl group, a substituted alkyl, an aryl group, a substituted aryl group, or a diamine. In some embodiments, L 6 In some embodiments, L 6 In some embodiments, L comprises a modified polyethylene glycol. 6 In some embodiments, L 6 In some embodiments, L 6comprises an aryl group or a substituted aryl group. In some embodiments, L 6 comprises a diamine (eg, a linking group comprising an alkylenediamine).

[0288] In some embodiments, L A is a first linker comprising: -(L 1 ) a -(L 2 ) b -(L 3 ) c -(L 4 ) d -(L 5 ) e -(L 6 ) f -, -(L 1 ) a -But-(T 1 -V 1 ) a - and -(L 2 ) b -But-(T 2 -V 2 ) b - and -(L 3 ) c -But-(T 3 -V 3 ) c - and -(L 4 ) d -But-(T 4 -V 4 ) d - and -(L 5 ) e -But-(T 5 -V 5 ) e - and -(L 6 ) f -But-(T 6 -V 6 ) f - and T 1 , T 2 , T 3 , T4 , T 5 , and T 6 is a tether group, if present, V 1 , V 2 , V 3 , V 4 , V 5 , and V 6 is, when present, a covalent bond or linking functional group; 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.

[0289] In certain embodiments, the sum of a, b, c, d, e, and f is 1 to 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 each 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.

[0290] As noted above, in certain embodiments, L 1 is attached to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety (e.g., as shown in formula (I) above). Thus, in certain embodiments, T 1is attached to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety (e.g., as shown in formula (I) above). In certain embodiments, V 1 is attached to the first drug or active agent. In certain embodiments, L 2 When present, T is attached to the first drug or active agent. 2 is attached to a first drug or active agent, if present, or V 2 When present, L is attached to the first drug or active agent. In certain embodiments, L 3 When present, T is attached to the first drug or active agent. 3 is attached to a first drug or active agent, if present, or V 3 When present, L is attached to the first drug or active agent. In certain embodiments, L 4 When present, T is attached to the first drug or active agent. 4 is attached to a first drug or active agent, if present, or V 4 When present, L is attached to the first drug or active agent. In certain embodiments, L 5 When present, T is attached to the first drug or active agent. 5 is attached to a first drug or active agent, if present, or V 5 When present, L is attached to the first drug or active agent. In certain embodiments, L 6 When present, T is attached to the first drug or active agent. 6 is attached to a first drug or active agent, if present, or V 6 When present, is attached to the first drug or active agent.

[0291] In certain embodiments, the conjugate of Formula (I) comprises a second linker, L B The second linker LB can be utilized to attach a second moiety of interest (e.g., a second drug or active agent) to the TF antibody via a conjugation moiety. B may be attached (e.g., covalently attached) to a conjugation moiety (e.g., as described herein). For example, a second linker, L B can link a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety to a second drug. The hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety can be linked to a second linker, L B (and thus a second drug) to the TF antibody.

[0292] For example, as shown in formula (I) above, L B is attached to the Ab via a conjugation moiety, such that the Ab is linked to a second linker L via a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety. B As mentioned above, the Ab is a TF antibody and therefore binds to L B is attached to the TF antibody via a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety, e.g., a linker L B is indirectly attached to the TF antibody via a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety.

[0293] The second linker L in the subject conjugates and compounds B Any convenient linker may be utilized. In certain embodiments, the second linker, L Bmay comprise a group selected from alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acylamino, alkylamido, substituted alkylamido, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, the second linker L B may comprise an alkyl or substituted alkyl group. In certain embodiments, the second linker, L B may comprise an alkenyl group or a substituted alkenyl group. In certain embodiments, the second linker, L B may comprise an alkynyl group or a substituted alkynyl group. In certain embodiments, the second linker, L B may comprise an alkoxy group or a substituted alkoxy group. In certain embodiments, the second linker, L B may comprise an amino group or a substituted amino group. In certain embodiments, the second linker, L B may contain a carboxyl group or a carboxyl ester group. In certain embodiments, the second linker L B may comprise an acylamino group. In certain embodiments, the second linker, L B may comprise an alkylamide group or a substituted alkylamide group. In certain embodiments, the second linker, L B may comprise an aryl group or a substituted aryl group. In certain embodiments, the second linker, L B may comprise a heteroaryl group or a substituted heteroaryl group. In certain embodiments, the second linker, L B may comprise a cycloalkyl or substituted cycloalkyl group. In certain embodiments, the second linker, L B may contain a heterocyclyl group or a substituted heterocyclyl group.

[0294] In certain embodiments, the second linker L Bmay comprise 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, copolymers of ethylene glycol and propylene glycol (e.g., the homopolymers and copolymers are unsubstituted or substituted at one end with an alkyl group), polyvinyl alcohol, polyvinyl ethyl ether, polyvinylpyrrolidone, combinations thereof, and the like. In certain embodiments, the polymer is a polyalkylene glycol. In certain embodiments, the polymer is polyethylene glycol. Other linkers can be used, as indicated in the conjugates and compounds described in more detail below.

[0295] In some embodiments, L B is a second linker described by the formula: -(L 7 ) g -(L 8 ) h -(L 9 ) i -(L 10 ) j -(L 11 ) k -(L 12 ) l -(L 13 ) m , L 7 , L 8 , L 9 , L 10 , L 11 , L 12 , and L 13 are each independently a linker subunit; and g, h, i, j, k, l, and m are each independently 0 or 1, with the proviso that at least one of g, h, i, j, k, l, and m is 1.

[0296] In certain embodiments, the sum of g, h, i, j, k, l, and m is 1 to 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 certain embodiments, g, h, i, j, and k are each 1 and l and m are each 0. In certain embodiments, g, h, i, and j are each 1 and k, l, and m are each 0. In certain embodiments, g, h, and i are each 1 and j, k, l, and m are each 0. In certain embodiments, g and h are each 1 and i, j, k, l, and m are each 0. In certain embodiments, g is 1 and h, i, j, k, l, and m are each 0. In certain embodiments, g, h, i, j, k, l, and m are each 0.

[0297] In certain embodiments, the linker subunit L 7 is attached to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety (e.g., as shown in formula (I) above). In certain embodiments, the linker subunit L 8 If present, the second drug or active agent W 2 In certain embodiments, the linker subunit L 9 If present, the second drug or active agent W 2 In certain embodiments, the linker subunit L 10 If present, the second drug or active agent W 2In certain embodiments, the linker subunit L 11 If present, the second drug or active agent W 2 In certain embodiments, the linker subunit L 12 If present, the second drug or active agent W 2 In certain embodiments, the linker subunit L 13 If present, the second drug or active agent W 2 is bonded to.

[0298] Any convenient linker subunit can be used as the second linker L B Linker subunits of interest include, but are not limited to, units of polymers (e.g., polyethylene glycol, polyethylene and polyacrylate, amino acid residue(s), carbohydrate-based 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 (when 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 comprising alkylenediamines).

[0299] In some embodiments, L 7 (when present) comprises polyethylene glycol, modified polyethylene glycol, an amino acid residue, an alkyl group, a substituted alkyl, an aryl group, a substituted aryl group, or a diamine. In some embodiments, L 7 In some embodiments, L 7 In some embodiments, L comprises a modified polyethylene glycol. 7In some embodiments, L 7 In some embodiments, L 7 comprises an aryl group or a substituted aryl group. In some embodiments, L 7 comprises a diamine (eg, a linking group comprising an alkylenediamine).

[0300] In some embodiments, L 8 (when present) comprises polyethylene glycol, modified polyethylene glycol, an amino acid residue, an alkyl group, a substituted alkyl, an aryl group, a substituted aryl group, or a diamine. In some embodiments, L 8 In some embodiments, L 8 In some embodiments, L comprises a modified polyethylene glycol. 8 In some embodiments, L 8 In some embodiments, L 8 comprises an aryl group or a substituted aryl group. In some embodiments, L 8 comprises a diamine (eg, a linking group comprising an alkylenediamine).

[0301] In some embodiments, L 9 (when present) comprises polyethylene glycol, modified polyethylene glycol, an amino acid residue, an alkyl group, a substituted alkyl, an aryl group, a substituted aryl group, or a diamine. In some embodiments, L 9 In some embodiments, L 9 In some embodiments, L comprises a modified polyethylene glycol. 9 In some embodiments, L 9 In some embodiments, L 9 comprises an aryl group or a substituted aryl group. In some embodiments, L 9comprises a diamine (eg, a linking group comprising an alkylenediamine).

[0302] In some embodiments, L 10 (when present) comprises polyethylene glycol, modified polyethylene glycol, an amino acid residue, an alkyl group, a substituted alkyl, an aryl group, a substituted aryl group, or a diamine. In some embodiments, L 10 In some embodiments, L 10 In some embodiments, L comprises a modified polyethylene glycol. 10 In some embodiments, L 10 In some embodiments, L 10 comprises an aryl group or a substituted aryl group. In some embodiments, L 10 comprises a diamine (eg, a linking group comprising an alkylenediamine).

[0303] In some embodiments, L 11 (when present) comprises polyethylene glycol, modified polyethylene glycol, an amino acid residue, an alkyl group, a substituted alkyl, an aryl group, a substituted aryl group, or a diamine. In some embodiments, L 11 In some embodiments, L 11 In some embodiments, L comprises a modified polyethylene glycol. 11 In some embodiments, L 11 In some embodiments, L 11 comprises an aryl group or a substituted aryl group. In some embodiments, L 11 comprises a diamine (eg, a linking group comprising an alkylenediamine).

[0304] In some embodiments, L 12(when present) comprises polyethylene glycol, modified polyethylene glycol, an amino acid residue, an alkyl group, a substituted alkyl, an aryl group, a substituted aryl group, or a diamine. In some embodiments, L 12 In some embodiments, L 12 In some embodiments, L comprises a modified polyethylene glycol. 12 In some embodiments, L 12 In some embodiments, L 12 comprises an aryl group or a substituted aryl group. In some embodiments, L 12 comprises a diamine (eg, a linking group comprising an alkylenediamine).

[0305] In some embodiments, L 13 (when present) comprises polyethylene glycol, modified polyethylene glycol, an amino acid residue, an alkyl group, a substituted alkyl, an aryl group, a substituted aryl group, or a diamine. In some embodiments, L 13 In some embodiments, L 13 In some embodiments, L comprises a modified polyethylene glycol. 13 In some embodiments, L 13 In some embodiments, L 13 comprises an aryl group or a substituted aryl group. In some embodiments, L 13 comprises a diamine (eg, a linking group comprising an alkylenediamine).

[0306] In some embodiments, L B is a second linker comprising: -(L 7 ) g -(L 8 ) h -(L 9 ) i -(L 10 ) j -(L11 ) k -(L 12 ) l -(L 13 ) m -, -(L 7 ) g -But-(T 7 -V 7 ) g - and -(L 8 ) h -But-(T 8 -V 8 ) h - and -(L 9 ) i -But-(T 9 -V 9 ) i - and -(L 10 ) j -But-(T 10 -V 10 ) j - and -(L 11 ) k -But-(T 11 -V 11 ) k - and -(L 12 ) l -But-(T 12 -V 12 ) l - and -(L 13 ) m -But-(T 13 -V 13 ) m - and T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 is a tether group, if present, V 7 , V 8 , V 9 , V 10 , V 11 , V 12 , and V13 is, when present, a covalent bond or linking functional group; 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.

[0307] In certain embodiments, the sum of g, h, i, j, k, l, and m is 1 to 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 certain embodiments, g, h, i, j, and k are each 1 and l and m are each 0. In certain embodiments, g, h, i, and j are each 1 and k, l, and m are each 0. In certain embodiments, g, h, and i are each 1 and j, k, l, and m are each 0. In certain embodiments, g and h are each 1 and i, j, k, l, and m are each 0. In certain embodiments, g is 1 and h, i, j, k, l, and m are each 0. In certain embodiments, g, h, i, j, k, l, and m are each 0.

[0308] As noted above, in certain embodiments, L 7 is attached to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety (e.g., as shown in formula (I) above). Thus, in certain embodiments, T 7is attached to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety (e.g., as shown in formula (I) above). In certain embodiments, V 7 is attached to a second drug or active agent. In certain embodiments, L 8 When present, T is attached to a second drug or active agent. 8 If present, is attached to a second drug or active agent, or V 8 When present, L is attached to a second drug or active agent. 9 When present, T is attached to a second drug or active agent. 9 If present, is attached to a second drug or active agent, or V 9 When present, L is attached to a second drug or active agent. 10 When present, T is attached to a second drug or active agent. 10 If present, is attached to a second drug or active agent, or V 10 When present, L is attached to a second drug or active agent. 11 When present, T is attached to a second drug or active agent. 11 If present, is attached to a second drug or active agent, or V 11 When present, L is attached to a second drug or active agent. 12 When present, T is attached to a second drug or active agent. 12 If present, is attached to a second drug or active agent, or V 12 When present, L is attached to a second drug or active agent. 13 When present, T is attached to a second drug or active agent. 13If present, is attached to a second drug or active agent, or V 13 If present, is attached to a second drug or active agent.

[0309] 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 With respect to T, any convenient tether group may be utilized in the subject linkers. 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 are covalent bonds, (C1 to 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-aminobenzyloxy (MABO), meta-aminobenzyloxycarbonyl (MABC), para-aminobenzyloxy (PABO), para-aminobenzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-aminobenzylamino (PABA), para-aminophenyl (PAP), para-hydroxyphenyl (PHP), acetal group, hydrazine, disulfide, and ester, wherein each w is an integer of 1 to 20, each n is an integer of 1 to 30, each p is an integer of 1 to 20, and each x is an integer of 1 to 12.

[0310] 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 In certain embodiments, (C1-C 12 ) alkyl is a straight-chain or branched 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, (C1 to C 12 ) alkyl is alkyl or substituted alkyl (e.g., C1-C 12 Alkyl, or C1-C 10 alkyl, or C1-C6 alkyl, or C1-C3 alkyl). 12 ) alkyl is C2 alkyl. For example, (C1-C 12 ) alkyl is alkylene or substituted alkylene (e.g., C1-C 12 Alkylene or C1-C 10alkylene, or C1-C6 alkylene, or C1-C3 alkylene). 12 ) alkyl is C1-alkylene (e.g., CH2). In some cases, (C1-C 12 ) alkyl is C2 alkylene (e.g., CH2CH2). In some cases, (C1-C 12 ) alkyl is C alkylene (e.g., CH2CH2CH2).

[0311] In certain embodiments, the substitution (C1-C 12 ) alkyl is a straight-chain 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, substituted (C1 to C 12 ) alkyl is a substituted alkyl (e.g., substituted C1-C 12 Alkyl or substituted C1-C 10 alkyl, or substituted C1-C6 alkyl, or substituted C1-C3 alkyl). 12 ) alkyl is a substituted C2 alkyl. For example, substituted (C1-C 12 ) alkyl is substituted alkylene (e.g., substituted C1-C 12 Alkylene or substituted C1-C 10 alkylene, or substituted C1-C6 alkylene, or substituted C1-C3 alkylene). 12 ) alkyl is a substituted C1-alkylene (e.g., a C1-alkylene substituted with —SO3H). In some cases, the substituted (C1-C 12 ) alkyl is a substituted C2 alkylene. In some cases, the substituted (C1-C 12 ) alkyl is a substituted C alkylene. For example, substituted (C-C 12 ) alkyl is 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 (e.g., -NHCO(PEG)7), but also C1-C substituted with -CONHCH2CH2SO3H groups. 12 It also includes alkylene (e.g., C3-alkylene), C1-C substituted with -NHCOCH2SO3H groups. 12 It may include alkylene (eg, C5-alkylene).

[0312] 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 ) includes aryl or substituted aryl. For example, the aryl can be phenyl. In some cases, the substituted aryl is substituted phenyl. Substituted phenyl is (C-C 12 ) alkyl, substituted (C1-C 12) 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 substituent comprises a cleavable moiety (e.g., an enzymatically cleavable moiety, e.g., a glycoside or glycoside derivative) described herein.

[0313] 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 heteroaryl or substituted heteroaryl, for example, triazolyl (e.g., 1,2,3-triazolyl). 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 cycloalkyl or substituted cycloalkyl. 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 substituted heterocyclyl. In some examples, a substituent on a substituted heteroaryl, substituted cycloalkyl, or substituted heterocyclyl comprises a cleavable moiety (e.g., an enzymatically cleavable moiety, e.g., a glycoside or glycoside derivative) described herein.

[0314] 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) w comprises one or more EDA moieties, e.g., 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 attached ethylenediamine (EDA) moieties can 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 by the following structure: [ka] (wherein y is an integer of 1 to 6, r is 0 or 1, and each R 12are 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, alkylamido, substituted alkylamido, 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 are independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. In certain embodiments, any two adjacent R 12 The groups can be cyclically linked, for example to form a piperazinyl ring. In certain embodiments, y is 1 and two adjacent R 12 In certain embodiments, y is 1 and the adjacent R 12 Groups are selected from hydrogen, alkyl (eg, methyl), and substituted alkyl (eg, lower alkyl-OH, such as ethyl-OH or propyl-OH).

[0315] 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 to herein as piperidine-4-amino, P4A). The 4AP moiety can be optionally 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 by the following structure: [ka] (In the formula, R 12 is 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, alkylamido, substituted alkylamido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl). 12 is a polyethylene glycol moiety. In certain embodiments, R 12 is a carboxy-modified polyethylene glycol.

[0316] In certain embodiments, R 12 is of the formula: (PEG) k which may 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 17is 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. 17 is COOH. In certain embodiments, R 17 is OH. In certain embodiments, R 17 However, it is OCH3.

[0317] 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 is a polyethylene glycol or modified polyethylene glycol linking unit. In certain embodiments, (PEG) n is described by the following structure: [ka] (wherein n 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 some cases, n is 2. In some cases, n is 3. In some cases, n is 6. In some cases, n is 12.

[0318] In certain embodiments, 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 ) but (AA) p where AA is an amino acid residue. Any convenient amino acid can be utilized. Amino acids of interest include, but are 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), 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 between 1 and 50, e.g., between 1 and 40, 1 and 30, 1 and 20, 1 and 12, or 1 and 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.

[0319] 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 includes amino acid analogs. Amino acid analogs include compounds similar in structure and / or overall 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-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) are one or more modified groups within the molecule. Such modifications may include, but are not limited to, substituting an atom (e.g., N) for a related atom (e.g., S), adding a group (e.g., methyl, hydroxyl, etc.) or atom (e.g., Cl, Br, etc.), deleting a group, substituting a covalent bond (e.g., a single bond for a double bond), or a combination thereof. For example, amino acid analogs may include alpha-hydroxy acids, alpha-amino acids, and the like. Examples of amino acid analogs include, but are not limited to, sulfoalanine, and the like.

[0320] 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 the formula -(CR 13 OH) x-, where x is 0 or x 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, or 12. In certain embodiments, x is 1. In certain embodiments, x is 2. In certain embodiments, R 13 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, alkylamido, substituted alkylamido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. 13 is hydrogen. In certain embodiments, R 13 is alkyl or substituted alkyl (e.g., C 1-6 Alkyl or C 1-6 Substituted alkyl, or C 1-4 Alkyl or C 1-4 Substituted alkyl, or C 1-3 Alkyl or C 1-3 In certain embodiments, R 13 is an alkenyl or substituted alkenyl (e.g., C 2-6 Alkenyl or C 2-6 substituted alkenyl, or C 2-4 Alkenyl or C 2-4 substituted alkenyl, or C 2-3 Alkenyl or C 2-3 In certain embodiments, R 13 is alkynyl or substituted alkynyl. In certain embodiments, R 13 is alkoxy or substituted alkoxy. In certain embodiments, R 13 is amino or substituted amino. In certain embodiments, R 13 is carboxyl or carboxyl ester. In certain embodiments, R13 is 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 sulfonyl. In certain embodiments, R 13 is thioalkoxy or substituted thioalkoxy. In certain embodiments, R 13 is an aryl or substituted aryl (e.g., C 5-8 Aryl or C 5-8 substituted aryl, e.g., C aryl or C substituted aryl, or C aryl or C substituted aryl). In certain embodiments, R 13 is heteroaryl or substituted heteroaryl (e.g., C 5-8 Heteroaryl or C 5-8 In certain embodiments, R is a C heteroaryl or a C substituted heteroaryl, such as a C heteroaryl or a C substituted heteroaryl. 13 is a cycloalkyl or substituted cycloalkyl (e.g., C 3-8 Cycloalkyl or C 3-8 Substituted cycloalkyl, e.g., C 3-6 Cycloalkyl or C 3-6 substituted cycloalkyl, or C 3-5 Cycloalkyl or C 3-5 In certain embodiments, R 13 is heterocyclyl or substituted heterocyclyl (e.g., C 3-8 Heterocyclyl or C 3-8 Substituted heterocyclyl, e.g., C 3-6 Heterocyclyl or C 3-6 substituted heterocyclyl, or C 3-5 Heterocyclyl or C 3-5 substituted heterocyclyl).

[0321] In certain embodiments, R 13is selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. In these embodiments, the alkyl, substituted alkyl, aryl, and substituted aryl are selected from R 13 The above is as described above.

[0322] 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 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.

[0323] 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 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).

[0324] In some embodiments, the tether group comprises a MABO group, which is depicted in the following structure: [ka]

[0325] In some embodiments, the tether group comprises an MABC group, which is described by the following structure: [ka]

[0326] In some embodiments, the tether group comprises a PABO group, which is depicted in the following structure: [ka]

[0327] In some embodiments, the tether group comprises a PABC group, which is described by the following structure: [ka]

[0328] In some embodiments, the tether group comprises a PAB group described by the following structure: [ka]

[0329] In some embodiments, the tether group comprises a PABA group described by the following structure: [ka]

[0330] In some embodiments, the tether group comprises a PAP group, which is described by the following structure: [ka]

[0331] In some embodiments, the tether group comprises a PHP group described by the following structure: [ka]

[0332] In certain embodiments, each R 14 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, alkylamido, substituted alkylamido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

[0333] In certain embodiments, R 14 is hydrogen. In certain embodiments, each R 14 is hydrogen. In certain embodiments, R 14 is alkyl or substituted alkyl (e.g., C 1-6 Alkyl or C 1-6 Substituted alkyl, or C 1-4 Alkyl or C 1-4 Substituted alkyl, or C 1-3 Alkyl or C 1-3 In certain embodiments, R 14 is an alkenyl or substituted alkenyl (e.g., C 2-6 Alkenyl or C 2-6 substituted alkenyl, or C 2-4 Alkenyl or C 2-4 substituted alkenyl, or C 2-3 Alkenyl or C 2-3 In certain embodiments, R 14 is alkynyl or substituted alkynyl. In certain embodiments, R 14 is alkoxy or substituted alkoxy. In certain embodiments, R 14is amino or substituted amino. In certain embodiments, R 14 is carboxyl or carboxyl ester. In certain embodiments, R 14 is 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 sulfonyl. In certain embodiments, R 14 is thioalkoxy or substituted thioalkoxy. In certain embodiments, R 14 is an aryl or substituted aryl (e.g., C 5-8 Aryl or C 5-8 substituted aryl, e.g., C aryl or C substituted aryl, or C aryl or C substituted aryl). In certain embodiments, R 14 is heteroaryl or substituted heteroaryl (e.g., C 5-8 Heteroaryl or C 5-8 In certain embodiments, R is a C heteroaryl or a C substituted heteroaryl, such as a C heteroaryl or a C substituted heteroaryl. 14 is a cycloalkyl or substituted cycloalkyl (e.g., C 3-8 Cycloalkyl or C 3-8 Substituted cycloalkyl, e.g., C 3-6 Cycloalkyl or C 3-6 substituted cycloalkyl, or C 3-5 Cycloalkyl or C 3-5 In certain embodiments, R 14 is heterocyclyl or substituted heterocyclyl (e.g., C 3-8 Heterocyclyl or C 3-8 Substituted heterocyclyl, e.g., C 3-6 Heterocyclyl or C 3-6 substituted heterocyclyl, or C 3-5 Heterocyclyl or C 3-5 substituted heterocyclyl).

[0334] In some embodiments of the MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP tether structures shown above, the phenyl ring can be substituted with one or more additional groups selected from halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acylamino, aminoacyl, alkylamido, substituted alkylamido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

[0335] 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 and one or more of T are each optionally substituted with a glycoside or glycoside derivative. For example, in some cases, T 1 , T 2 , T 3 , T 4 , T 5 , and T 6 are each optionally substituted with a glycoside. 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 are each 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.

[0336] In certain embodiments, the MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP tether structures shown above can 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 can 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, e.g., a hydrogen atom of PABC is substituted with a glycoside (e.g., glucuronide, galactoside, glucoside, mannoside, fucoside, O-GlcNAc, and O-GalNAc).

[0337] For example, in some embodiments, the glycoside or glycoside derivative can be selected from the following structures: [ka]

[0338] Bonding functional group 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 13With respect to V, any convenient linking functional group may be utilized in the subject linkers. Binding functional groups of interest include, but are not limited to, amino, carbonyl, amido, oxycarbonyl, carboxy, sulfonyl, sulfoxide, sulfonylamino, aminosulfonyl, thio, oxy, phospho, phosphoramidate, thiophosphoramidate, and the like. 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 are each independently a covalent bond, -CO-, or -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 and -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.

[0339] In some embodiments, each R 15is 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, alkylamido, substituted alkylamido, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

[0340] In certain embodiments, R 15 is hydrogen. In certain embodiments, each R 15 is hydrogen. In certain embodiments, R 15 is alkyl or substituted alkyl (e.g., C 1-6 Alkyl or C 1-6 Substituted alkyl, or C 1-4 Alkyl or C 1-4 Substituted alkyl, or C 1-3 Alkyl or C 1-3 In certain embodiments, R 15 is an alkenyl or substituted alkenyl (e.g., C 2-6 Alkenyl or C 2-6 substituted alkenyl, or C 2-4 Alkenyl or C 2-4 substituted alkenyl, or C 2-3 Alkenyl or C 2-3 In certain embodiments, R 15 is alkynyl or substituted alkynyl. In certain embodiments, R 15 is alkoxy or substituted alkoxy. In certain embodiments, R 15 is amino or substituted amino. In certain embodiments, R 15 is carboxyl or carboxyl ester. In certain embodiments, R 15 is 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 sulfonyl. In certain embodiments, R 15 is thioalkoxy or substituted thioalkoxy. In certain embodiments, R 15 is an aryl or substituted aryl (e.g., C 5-8 Aryl or C 5-8 substituted aryl, e.g., C aryl or C substituted aryl, or C aryl or C substituted aryl). In certain embodiments, R 15 is heteroaryl or substituted heteroaryl (e.g., C 5-8 Heteroaryl or C 5-8 In certain embodiments, R is a C heteroaryl or a C substituted heteroaryl, such as a C heteroaryl or a C substituted heteroaryl. 15 is a cycloalkyl or substituted cycloalkyl (e.g., C 3-8 Cycloalkyl or C 3-8 Substituted cycloalkyl, e.g., C 3-6 Cycloalkyl or C 3-6 substituted cycloalkyl, or C 3-5 Cycloalkyl or C 3-5 In certain embodiments, R 15 is heterocyclyl or substituted heterocyclyl (e.g., C 3-8 Heterocyclyl or C 3-8 Substituted heterocyclyl, e.g., C 3-6 Heterocyclyl or C 3-6 substituted heterocyclyl, or C 3-5 Heterocyclyl or C 3-5 substituted heterocyclyl).

[0341] In certain embodiments, each R 15are 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 are independently selected from R 15 The above is as described above.

[0342] As noted above, in some embodiments, L A But, -(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 -, wherein a, b, c, d, e, and f are each independently 0 or 1, and at least one of a, b, c, d, e, and f is 1.

[0343] In some embodiments, the first linker L A In: T 1 However, (C1-C 12 ) alkyl and substituted (C1-C 12 ) alkyl; T 2 , T 3 , T 4 , T 5 , and T 6are respectively and independently (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-aminopiperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, acetal groups, disulfides, hydrazines, and esters; V 1 , V 2 , V 3 , V 4 , V 5 , and V 6 are each independently a covalent bond, -CO-, or -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 selected from -SO2- and -P(O)OH-, and q is an integer of 1 to 6; (PEG) n but, [ka] and n is an integer from 1 to 30; EDA is an ethylenediamine moiety having the following structure: [ka] y is an integer from 1 to 6, and r is 0 or 1; 4-aminopiperidine (4AP) [ka] and AA is an amino acid residue, and p is an integer of 1 to 20; Each R 12 are independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, and any two adjacent R 12 The groups may be cyclically linked to form a piperazinyl ring, Each R 13 is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl; Each R 15 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.

[0344] In some embodiments, L A but includes: -(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, 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; T 1 , T 2 , T 3 , T 4 , T 5 , and T 6 are each independently 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-aminopiperidine (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 group, hydrazine, disulfide, and ester; EDA is an ethylenediamine moiety; PEG is polyethylene glycol; AA is an amino acid residue or amino acid analog; each w is an integer of 1 to 20; each n is an integer of 1 to 30; each p is an integer of 1 to 20; and each x is an integer of 1 to 12; V 1 , V 2 , V 3 , V 4 , V 5 , and V 6 are each independently a covalent bond, -CO-, or -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 and -SO2-, and -P(O)OH-, 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 15is 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.

[0345] L A In some embodiments of T 1 However, (C1-C 12 ) alkyl and substituted (C1-C 12 ) alkyl; T 2 , T 3 , T 4 , T 5 , and T 6 are each independently 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-aminopiperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, acetal groups, hydrazines, and esters; V 1 , V 2 , V 3 , V 4 , V 5 , and V 6 are each independently a covalent bond, -CO-, or -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 selected from the group consisting of SO2-, and -P(O)OH-; (PEG) n but, [ka] and n is an integer from 1 to 30; EDA is an ethylenediamine moiety having the following structure: [ka] y is an integer of 1 to 6, and r is an integer of 0 to 1; 4-aminopiperidine (4AP) [ka] and Each R 12 are independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, and any two adjacent R 12 The groups may be cyclically linked to form a piperazinyl ring, a, b, c, and d are each 1; e and f are 0.

[0346] In some embodiments, T 1 , T 2 , T 3 , T 4 , T 5 , and T 6 are each optionally substituted with a glycoside.

[0347] In some embodiments, MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP are each optionally substituted with a glycoside.

[0348] In some embodiments, the glycoside is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.

[0349] In certain embodiments, 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 is selected from: T 1 However, (C1-C 12 ) alkyl, and V 1 is -CO-, T 2 However, (AA) p and V 2 But it doesn't exist. T 3 But PABC, V 3 But it doesn't exist, p is an integer from 1 to 10, d, e, and f are each 0, or T 1 However, (C1-C 12 ) alkyl, and V 1 is -CONH-, T 2 However, (PEG) n and V 2 is -CO-, T 3 However, (AA) p and V 3 But it doesn't exist, T 4 But PABC, V 4 But it doesn't exist, p is an integer from 1 to 10, e and f are each 0, or T 1 However, (C1-C 12 ) alkyl, and V 1 is -CO-, T 2 is an amino acid analog, and V 2is -NH-, T 3 However, (PEG) n and V 3 is -CO-, T 4 However, (AA) p and V 4 But it doesn't exist, T 5 But PABC, V 5 But it doesn't exist, p is an integer from 1 to 10, f is 0, or T 1 However, (C1-C 12 ) alkyl, and V 1 is -CONH-, T 2 However, (PEG) n and V 2 is -CO-, T 3 However, (AA) p and V 3 But it doesn't exist, T 4 But PABC, V 4 But it doesn't exist, p is an integer from 1 to 10, e and f are each 0, or T 1 However, (C1-C 12 ) alkyl, and V 1 is -CONH-, T 2 is replaced by (C1-C 12 ) alkyl, and V 2 is -CO-, T 3 However, (AA) p and V 3 But it doesn't exist, T 4 But PABC, V 4 But it doesn't exist, p is an integer from 1 to 10, e and f are each 0, or T 1 However, (C1-C12 ) alkyl, and V 1 is -CONH-, T 2 However, (PEG) n and V 2 is -CO-, T 3 However, (AA) p and V 3 But it doesn't exist, T 4 However, PABA and V 4 is -CO-, T 5 However, (C1-C 12 ) alkyl, and V 5 But it doesn't exist, p is an integer from 1 to 10, f is 0, or T 1 However, (C1-C 12 ) alkyl, and V 1 is -CO-, T 2 But it is 4AP, V 2 is -CO-, T 3 However, (C1-C 12 ) alkyl, and V 3 is -CO-, T 4 However, (AA) p and V 4 But it doesn't exist, T 5 But PABC, V 5 But it doesn't exist, p is an integer from 1 to 10, f is 0, or T 1 However, (C1-C 12 ) alkyl, and V 1 is -CO-, T 2 But it is 4AP, V 2 is -CO-, T 3 However, (C1-C 12 ) alkyl, and V 3is -O-, T 4 However, (C1-C 12 ) alkyl, and V 4 is -CO-, T 5 However, (AA) p and V 5 But it doesn't exist, p is an integer from 1 to 10, T 6 But PABC, V 6 does not exist, or T 1 However, (C1-C 12 ) alkyl, and V 1 is -CO-, T 2 is an amino acid analog, and V 2 But it doesn't exist, T 3 However, (AA) p and V 3 But it doesn't exist, T 4 But PABC, V 4 But it doesn't exist, p is an integer from 1 to 10, e and f are each 0, or T 1 However, (C1-C 12 ) alkyl, and V 1 is -CONH-, T 2 However, (PEG) n and V 2 is -CONH-, T 3 is replaced by (C1-C 12 ) alkyl, and V 3 is -CO-, T 4 However, (AA) p and V 4 But it doesn't exist, T 5 But PABC, V 5 But it doesn't exist, p is an integer from 1 to 10, f is 0, or T 1 However, (C1-C 12 ) alkyl, and V 1 is -CO-, T 2 However, (AA) p and V 2 is -NH-, T 3 However, (PEG) n and V 3 is -CO-, T 4 However, (AA) p and V 4 But it doesn't exist, T 5 But PABC, V 5 But it doesn't exist, p is an integer from 1 to 10, f is 0, or T 1 However, (C1-C 12 ) alkyl, and V 1 is -CONH-, T 2 However, (PEG) n and V 2 is -CO-, T 3 However, (AA) p and V 3 But it doesn't exist, T 4 is PAP, and V 4 is -C(O)O-, p is an integer from 1 to 10, e and f are each 0, or T 1 However, (C1-C 12 ) alkyl, and V 1 is -CONH-, T 2 is replaced by (C1-C 12 ) alkyl, and V 2 is -CO-, T 3 However, (AA) p and V 3But it doesn't exist, T 4 But PABC, V 4 But it doesn't exist, p is an integer from 1 to 10, e and f are each 0, or T 1 However, (C1-C 12 ) alkyl, and V 1 is -CONH-, T 2 is replaced by (C1-C 12 ) alkyl, and V 2 is -CO-, T 3 But PABC, V 3 But it doesn't exist, d, e, and f are each 0.

[0350] In certain embodiments, the first linker L A is attached to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety, and a first linker L A The right side of the above linker structure is attached to a first drug or active agent.

[0351] As noted 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-, wherein g, h, i, j, k, l, and m are each independently 0 or 1, and at least one of g, h, i, j, k, l, and m is 1.

[0352] In some embodiments, the second linker L B In: T 7 However, (C1-C 12 ) alkyl and substituted (C1-C 12 ) alkyl; T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 However, each independently, 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-aminopiperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, acetal groups, disulfides, hydrazines, and esters; V 7 , V 8 , V 9 , V 10 , V 11 , V 12 and V 13 are each independently a covalent bond, -CO-, or -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 -, -NR15 -SO2- and -P(O)OH-, and q is an integer from 1 to 6; (PEG) n but, [ka] and n is an integer from 1 to 30; EDA is an ethylenediamine moiety having the following structure: [ka] y is an integer from 1 to 6, and r is 0 or 1; 4-aminopiperidine (4AP) [ka] and AA is an amino acid residue, and p is an integer of 1 to 20; Each R 12 are independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, and any two adjacent R 12 The groups may be cyclically linked to form a piperazinyl ring, Each R 13 is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl; Each R 15 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.

[0353] T 7 , T 8 , T 9 , T 10 , T 11 , T 12 and T 13 Any convenient tether group to T may be utilized. For example,1 , T 2 , T 3 , T 4 , T 5 , and T 6 In connection with any of the above tether groups, the tether group T 7 , T 8 , T 9 , T 10 , 11 , T 12 , and T 13 may be used for

[0354] V 7 , V 8 , V 9 , V 10 , V 11 , V 12 and V 13 Any convenient linking functionality to V may be used. For example, 1 , V 2 , V 3 , V 4 , V 5 , and V 6 In connection with any of the above linking functional groups, the linking functional group V 7 , V 8 , V 9 , V 10 , V 11 , V 12 , and V 13 can be used for.

[0355] In certain embodiments, each R 13 are independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. In these embodiments, alkyl, substituted alkyl, aryl, and substituted aryl are independently selected from R 13 The above is as described above.

[0356] In certain embodiments, each R 15are 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 are independently selected from R 15 In these embodiments, the various possible substituents are as described above for R 15 The above is as described above.

[0357] Second linker L B In certain embodiments, the tether group T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 is optionally substituted with a glycoside or glycoside derivative, respectively. In certain embodiments, the glycoside or glycoside derivative is selected from glucuronide, galactoside, glucoside, mannoside, fucoside, O-GlcNAc, and O-GalNAc.

[0358] Second linker L BIn certain embodiments, the MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP tether structures shown above can 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 can 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.

[0359] In some embodiments, T 7 , T 8 , T 9 , T 10 , T 11 , T 12 , and T 13 are each optionally substituted with a glycoside.

[0360] In some embodiments, MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP are each optionally substituted with a glycoside.

[0361] In some embodiments, the glycoside is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.

[0362] L B In some embodiments of g, h, i, j, and k are each 1; l and m are 0, T 7 is a covalent bond, T 8 , T 9 , T 10 , T 11 , and T 12 are each independently 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-aminopiperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, acetal groups, hydrazines, and esters; V 7 , V 8 , V 9 , V 10 , V 11 , and V 12 are each independently a covalent bond, -CO-, or -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 SO2-, and -P(O)OH-; (PEG) n but, [ka] and n is an integer from 1 to 30; EDA is an ethylenediamine moiety having the following structure: [ka] y is an integer of 1 to 6, and r is an integer of 0 to 1; 4-aminopiperidine (4AP) [ka] and Each R 12 are independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, and any two adjacent R 12 The groups may be cyclically linked to form a piperazinyl ring.

[0363] 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 are each optionally substituted with a glycoside.

[0364] In some embodiments, MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP are each optionally substituted with a glycoside.

[0365] In some embodiments, the glycoside is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.

[0366] In certain embodiments, 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 is selected from the following: T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12) alkyl, and V 8 is -CO-, T 9 However, (AA) p and V 9 But it doesn't exist, T 10 But PABC, V 10 But it doesn't exist, k, l, and m are each 0, or T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CONH-, T 9 However, (PEG) n and V 9 is -CO-, T 10 However, (AA) p and V 10 But it doesn't exist, T 11 But PABC, V 11 But it doesn't exist, l and m are each 0, or T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CO-, T 9 is an amino acid analog, and V 9 is -NH-, T 10 However, (PEG) n and V 10 is -CO-, T 11 However, (AA) p and V 11 But it doesn't exist, T 12 But PABC, V 12 But it doesn't exist, m is 0, or T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CONH-, T 9 However, (PEG) n and V 9 is -CO-, T 10 However, (AA) p and V 10 But it doesn't exist, T 11 But PABC, V 11 But it doesn't exist, l and m are each 0, or T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CONH-, T 9 is replaced by (C1-C 12 ) alkyl, and V 9 is -CO-, T 10 However, (AA) p and V 10 But it doesn't exist, T 11 But PABC, V 11 But it doesn't exist, l and m are each 0, or T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CONH-, T 9 However, (PEG) n and V 9 is -CO-, T 10 However, (AA) p and V 10 But it doesn't exist, T 11 However, PABA and V 11 is -CO-, T 12 However, (C1-C 12 ) alkyl, and V 12 But it doesn't exist, m is 0, or T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CO-, T 9 But it is 4AP, V 9 is -CO-, T 10 However, (C1-C 12 ) alkyl, and V 10 is -CO-, T 11 However, (AA) p and V 11 But it doesn't exist, T 12 But PABC, V 12 But it doesn't exist, m is 0, or T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CO-, T 9 But it is 4AP, V 9 is -CO-, T 10 However, (C1-C 12 ) alkyl, and V 10 is -O-, T 11 However, (C1-C 12) alkyl, and V 11 is -CO-, T 12 However, (AA) p and V 12 But it doesn't exist, T 13 PABC and V 13 does not exist, or T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CO-, T 9 is an amino acid analog, and V 9 But it doesn't exist, T 10 However, (AA) p and V 10 But it doesn't exist, T 11 But PABC, V 11 But it doesn't exist, l and m are each 0, or T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CONH-, T 9 However, (PEG) n and V 9 is -CONH-, T 10 is replaced by (C1-C 12 ) alkyl, and V 10 is -CO-, T 11 However, (AA) p and V 11 But it doesn't exist, T 12 But PABC, V 12 But it doesn't exist, m is 0, or T7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CO-, T 9 However, (AA) p and V 9 is -NH-, T 10 However, (PEG) n and V 10 is -CO-, T 11 However, (AA) p and V 11 But it doesn't exist, T 12 But PABC, V 12 But it doesn't exist, m is 0, or T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CONH-, T 9 However, (PEG) n and V 9 is -CO-, T 10 However, (AA) p and V 10 But it doesn't exist, T 11 is PAP, and V 11 is -C(O)O-, l and m are each 0, or T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CO-, T 9 However, (AA) pand V 9 But it doesn't exist, T 10 But PABC, V 10 But it doesn't exist, T 11 is PAP, and V 11 is -C(O)O-, l and m are each 0, or T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CONH-, T 9 is replaced by (C1-C 12 ) alkyl, and V 9 is -CO-, T 10 But PABC, V 10 But it doesn't exist, k, l, and m are each 0, or T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 But it doesn't exist, T 9 is heteroaryl, and V 9 But it doesn't exist, T 10 However, (C1-C 12 ) alkyl, and V 10 is -CONH-, T 11 However, (PEG) n and V 11 is -CO-, l and m are each 0, or T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12) alkyl, and V 8 But it doesn't exist, T 9 is heteroaryl, and V 9 But it doesn't exist, T 10 However, (C1-C 12 ) alkyl, and V 10 is -CONH-, T 11 is replaced by (C1-C 12 ) alkyl, and V 11 is -CO-, T 12 However, (AA) p and V 12 But it doesn't exist, T 13 PAB and V 13 does not exist, or T 7 But it doesn't exist, V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 But it doesn't exist, T 9 is heteroaryl, and V 9 But it doesn't exist, T 10 However, (C1-C 12 ) alkyl, and V 10 is -CONH-, T 11 is replaced by (C1-C 12 ) alkyl, and V 11 is -CO-, T 12 However, (AA) p and V 12 But it doesn't exist, T 13 PABC and V 13 But it doesn't exist.

[0367] In certain embodiments, the second linker L Bis attached to a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety, and a second linker, L B The right side of the above linker structure is attached to a second drug or active agent.

[0368] In certain embodiments, the conjugate is an antibody-drug conjugate, as described above, in which the TF antibody and the drug are joined by a linker. In some cases, the linker m (e.g., L A and / or L B ) is a cleavable linker. A cleavable linker is a linker that includes one or more cleavable moieties, where the cleavable moieties include one or more bonds that can dissociate under certain conditions, thereby separating the cleavable linker into two or more separable moieties; for example, the cleavable moieties may include one or more covalent bonds that can dissociate or decompose under certain conditions, separating the cleavable linker into two or more moieties. Thus, the linker included in the antibody-drug conjugate can be a cleavable linker such that, under appropriate conditions, the cleavable linker is cleaved, separating or releasing the drug from the antibody at the desired target site of action of the drug.

[0369] In some cases, a cleavable linker comprises two cleavable moieties (e.g., a first cleavable moiety and a second cleavable moiety). The cleavable moieties can be configured such that cleavage of both cleavable moieties is required to separate or release the drug from the TF antibody at the desired target site of action of the drug. For example, cleavage of the cleavable linker can be achieved by first cleaving one of the two cleavable moieties and then cleaving the other of the two cleavable moieties. In certain embodiments, the cleavable linker comprises a first cleavable moiety and a second cleavable moiety that prevents cleavage of the first cleavable moiety. "Prevents cleavage" means that the presence of an uncleaved second cleavable moiety reduces or substantially inhibits the likelihood of cleavage of the first cleavable moiety, thereby substantially reducing or preventing the amount of cleavable linker. For example, the presence of an uncleaved second cleavable moiety can prevent cleavage of the first cleavable moiety. If the presence of the second cleavable moiety prevents cleavage of the first cleavable moiety, then the amount of drug from the antibody is greatly reduced or its release is prevented, For example, premature release of the drug from the antibody can be greatly reduced or prevented until the antibody-drug conjugate reaches or near the desired target site of action of the drug.

[0370] In some cases, because the second cleavable moiety prevents the cleavage of the first cleavable moiety, cleavage of the cleavable linker can be achieved by first cleaving the second cleavable moiety and then cleaving the first cleavable moiety. Cleavage of the second cleavable moiety reduces or eliminates the interference with cleavage of the first cleavable moiety, thereby enabling cleavage of the first cleavable moiety. Cleavage of the first cleavable moiety can cause the cleavable linker to dissociate or separate into two or more moieties, as described above, thereby releasing the drug from the antibody-drug conjugate. In some cases, cleavage of the first cleavable moiety does not substantially occur in the presence of an uncleaved second cleavable moiety. Substantially means that about 10% or less of cleavage of the first cleavable portion occurs in the presence of an uncleaved second cleavable portion, for example, about 9% or less, or about 8% or less, or about 7% or less, or about 6% or less, or about 5% or less, or about 4% or less, or about 3% or less, or about 2% or less, or about 1% or less, or about 0.5% or less, or about 0.1% or less of cleavage of the first cleavable portion occurs in the presence of an uncleaved second cleavable portion.

[0371] In other words, the second cleavable moiety can protect the first cleavable moiety from cleavage. For example, the presence of an uncleaved second cleavable moiety protects the first cleavable moiety from cleavage, thereby significantly reducing or preventing premature release of the drug from the antibody until the antibody-drug conjugate reaches or near the intended target site of action of the drug. Thus, cleavage of the second cleavable moiety exposes the first cleavable moiety (e.g., deprotects the first cleavable moiety), allowing cleavage of the first cleavable moiety, resulting in cleavage of the cleavable linker, and then separating or releasing the drug from the antibody at the desired target site of action of the drug, as described above. In certain cases, cleavage of the second cleavable moiety exposes the first cleavable moiety to subsequent cleavage, but cleavage of the second cleavable moiety does not, in itself, result in cleavage of the cleavable linker (i.e., cleavage of the first cleavable site is still required to cleave the cleavable linker).

[0372] Each cleavable moiety included in the cleavable linker may be an enzymatically cleavable moiety. For example, the first cleavable moiety may be a first enzymatically cleavable moiety, and the second cleavable moiety may be a second enzymatically cleavable moiety. An enzymatically cleavable moiety is a cleavable moiety that can be separated into two or more moieties by the enzymatic action of an enzyme. The enzymatically cleavable moiety may be any moiety (e.g., but not limited to, an ester, a peptide, a glycoside, etc.) that can be cleaved through the enzymatic action of an enzyme. In some cases, the enzyme that cleaves the enzymatically cleavable moiety is present at the desired target site of action (e.g., the desired target site of action of the drug to be released from the antibody-drug conjugate). In some cases, the enzyme that cleaves the enzymatically cleavable moiety is not present in significant amounts in other regions (e.g., whole blood, plasma, serum). Thus, when the enzymatically cleavable moiety is cleaved, substantial cleavage occurs at the desired site of action, but cleavage can be controlled so that it does not occur significantly in other regions or before the antibody-drug conjugate reaches the desired site of action.

[0373] For example, as described herein, the antibody-drug conjugates of the present disclosure can be used to treat cancer (e.g., to deliver a cancer therapeutic to a desired site of action where cancer cells are present). In some cases, enzymes (e.g., esterases, which cleave ester bonds, and glycosidases, which cleave glycosidic bonds) can be biomarkers for cancer that are overexpressed in cancer cells. The overexpression, and thus localization, of specific enzymes in cancer can be used in conjunction with an enzymatically cleavable moiety included in the cleavable linker of an antibody-drug conjugate of the present disclosure to specifically release a drug at a desired site of action (i.e., the site of the 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 that is overexpressed in cancer cells. For example, the enzyme can be an esterase. Thus, in some cases, the enzymatically cleavable moiety is a cleavable moiety (e.g., an ester) that can be cleaved by an esterase enzyme. In some cases, the enzyme can be a glycosidase. Thus, in some cases, the enzymatically cleavable moiety is a cleavable moiety (e.g., a glycoside or glycoside derivative) that can be cleaved with a glycosidase enzyme.

[0374] In certain embodiments, the enzymatically cleavable moiety is an ester bond. For example, the first cleavable moiety (i.e., the cleavable moiety protected from premature cleavage by the second cleavable moiety) described above comprises an ester. The presence of the uncleaved second cleavable moiety protects the first cleavable moiety (ester) from cleavage by an esterase enzyme, 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 action of the drug. In some cases, a portion of the linker adjacent to the first cleavable moiety is bound to or includes a substituent that includes a second cleavable moiety. In some cases, the second cleavable moiety includes a glycoside or a glycoside derivative.

[0375] In some embodiments, the enzymatically cleavable moiety is a sugar moiety, such as a glycoside (or glyosyl) or glycoside derivative. In some cases, the glycoside or glycoside derivative may promote increased hydrophilicity of the cleavable linker compared to a cleavable linker that does not contain a glycoside or glycoside derivative. The glycoside or glycoside derivative may be any glycoside or glycoside derivative that is suitable for use in a cleavable linker and can be cleaved by the enzymatic action of an enzyme. For example, the second cleavable moiety (i.e., the cleavable moiety that protects the first cleavable moiety from premature cleavage) may be a glycoside or glycoside derivative. For example, in some embodiments, the first cleavable moiety comprises an ester and the second cleavable moiety comprises a glycoside or glycoside derivative. In certain embodiments, the second cleavable moiety is a glycoside or glycoside derivative selected from glucuronide, galactoside, glucoside, mannoside, fucoside, O-GlcNAc, and O-GalNAc. In some cases, the second cleavable moiety is a glucuronide. In some cases, the second cleavable moiety is a galactoside. In some cases, the second cleavable moiety is a glucoside. In some cases, the second cleavable moiety is a mannoside. In some cases, the second cleavable moiety is a fucoside. In some cases, the second cleavable moiety is O-GlcNAc. In some cases, the second cleavable moiety is O-GalNAc.

[0376] The glycoside or glycoside derivative can be covalently attached to the cleavable linker via a glycosidic bond. The glycosidic bond can be attached to the cleavable linker via various types of bonds (e.g., but not limited to, an O-glycosidic bond (O-glycoside), an N-glycosidic bond (glycosylamine), an S-glycosidic bond (thioglycoside), or a C-glycosidic bond (C-glycoside or C-glycosyl). In some cases, the glycosidic bond is an O-glycosidic bond (O-glycoside). In some cases, the glycoside or glycoside derivative can be enzymatically cleaved from the cleavable linker to which it is attached (e.g., via enzyme-mediated hydrolysis of the glycosidic bond). The glycoside or glycoside derivative can be removed or cleaved from the cleavable linker by any convenient enzyme capable of cleaving (hydrolyzing) the glycosidic bond connecting the glycoside or glycoside derivative to the cleavable linker. Examples of enzymes that can be used to mediate the cleavage (hydrolysis) of the glycosidic bond connecting the glycoside or glycoside derivative to the cleavable linker are glycosidases (e.g., glucuronidase, galactosidase, glucosidase, mannosidase, fucosidase, etc.). Other suitable enzymes can also be used to mediate the cleavage (hydrolysis) of the glycosidic bond connecting the glycoside or glycoside derivative to the cleavable linker. In some cases, the enzyme that mediates the cleavage (hydrolysis) of the glycosidic bond connecting the glycoside or glycoside derivative to the cleavable linker is found at or near the desired site of action of the drug of the antibody-drug conjugate. For example, the enzyme can be a lysosomal enzyme (e.g., a lysosomal glycosidase) found within the cell at or near the desired site of action of the drug of the antibody drug conjugate. In some cases, the enzyme is found at or near the target site where the enzyme that mediates cleavage of the first cleavable moiety is found.

[0377] In some embodiments, the TF-ADC has formula (I): [ka] (In the formula, Ab represents an antibody that binds to TF; Z 1 , Z 2 , and Z 4 However, each independently, CR 4 and Z 3 But CL B -W 2 and R 1 , R 2 , R 3 , and R 4 are hydrogen and (C1-C 12 ) alkyl; L A is the first linker, T 1 However, (C1-C 12 ) alkyl, and V 1 is -CONH-, T 2 is replaced by (C1-C 12 ) alkyl, and V 2 is -CO-, T 3 However, (AA) p where p is an integer from 1 to 20, and V 3 is a covalent bond, T 4 But PABC, V 4 is a covalent bond, a, b, c, and d are each 1; e and f are each 0; L B is the second linker, T 7 is a covalent bond, and V 7 is -NHCO-, T 8 However, (C1-C 12 ) alkyl, and V 8 is -CONH-, T 9 is replaced by (C1-C12 ) alkyl, and V 9 is -CO-, T 10 However, (AA) p where p is an integer from 1 to 20, and V 10 is a covalent bond, T 11 But PABC, V 11 is a covalent bond, h, i, j, and k are each 1; l and m are each 0; s is an integer from 1 to 10, W 1 is the first drug, W 2 is the second drug).

[0378] In some embodiments, W 1 and W 2 One or both of the is a camptothecin analog (e.g., belotecan).

[0379] In some embodiments, the TF-ADC has formula (I): [ka] (In the formula, Ab represents an antibody that binds to TF; Z 1 , Z 2 , and Z 4 However, each independently, CR 4 and Z 3 But CL B -W 2 and R 1 , R 2 , R 3 , and R 4 are hydrogen and (C1-C 12 ) alkyl; L A is the linker, T 1 is (C1-C6) alkyl, and V1 is -CONH-, T 2 But -NHCO(PEG) k where k is an integer from 2 to 10, and V 2 is -CO-, T 3 is (AA)2 and V 3 is a covalent bond, T 4 is a glycosidic-substituted PABC, and V 4 is a covalent bond, a, b, c, and d are each 1; e and f are each 0; L B is the linker, T 7 is a covalent bond, and V 7 is -NHCO-, T 8 is (C1-C6) alkyl, and V 8 is -CONH-, T 9 But -NHCO(PEG) k where k is an integer from 2 to 10, and V 9 is -CO-, T 10 is (AA)2 and V 10 is a covalent bond, T 11 is a glycosidic-substituted PABC, and V 11 is a covalent bond, h, i, j, and k are each 1; l and m are each 0; s is an integer from 1 to 10, W 1 is the first drug, W 2 is the second drug).

[0380] In some embodiments, T 4 and T 11In some embodiments, one or both PABCs in T are substituted with a glucuronide. 1 and T 8 In some embodiments, one or both of T 2 and T 9 One or both of the groups may be -NHCO(PEG) k and k is an integer from 5 to 10. In some embodiments, W 1 and W 2 One or both of the is a camptothecin analog (e.g., belotecan).

[0381] In some embodiments, the TF-ADCs disclosed herein comprise any payload, any linker, or any linker-payload disclosed in U.S. Patent Application No. 2022-0241423 and International Publication No. WO2022187370, each of which is incorporated herein by reference in its entirety.

[0382] In some embodiments, the TF-ADC has formula (II): [ka] (In the formula, Ab represents an antibody that binds to TF; and s is an integer from 1 to 10.

[0383] In some embodiments, s is an integer from 1 to 4.

[0384] Any of the chemicals, linkers, and conjugation moieties described in the structures above can be adapted for use in the subject compounds and conjugates.

[0385] Additional disclosure regarding hydrazinyl-indolyl and hydrazinyl-pyrrolo-pyridinyl compounds and methods for producing the conjugates can be found in U.S. Pat. Nos. 9,310,374 and 9,493,413, the disclosures of each of which are incorporated herein by reference.

[0386] Tissue factor (TF) antibody As noted above, the subject conjugates include an antibody (Ab) that binds to TF. The amino acid sequence of the antibody can be modified to include 2-formylglycine (fGly) residues. As used herein, amino acids may be referred to by their standard name, their standard three-letter abbreviation, and / or their standard one-letter abbreviation (e.g., alanine or Ala or A, cysteine ​​or Cys or C, aspartic acid or Asp or D, glutamic acid or Glu or E, phenylalanine or Phe or F, glycine or Gly or G, histidine or His or H, isoleucine or Ile or I, lysine or Lys or K, leucine or Leu or L, methionine or Met or M, asparagine or Asn or N, proline or Pro or P, glutamine or Gln or Q, arginine or Arg or R, serine or Ser or S, threonine or Thr or T, valine or Val or V, tryptophan or Trp or W, and tyrosine or Tyr or Y).

[0387] The TF-ADCs described herein comprise a drug and a TF antibody conjugated thereto. In some embodiments, a TF antibody refers to an antibody that specifically binds to TF (e.g., a TF protein, a TF polypeptide, a TF polypeptide fragment, a TF peptide, or a TF epitope). In some embodiments, the TF antibody is a human or humanized antibody (e.g., comprising a human constant region) that binds to TF. In some embodiments, the TF antibody can bind to TF expressed on the surface of mammalian (e.g., human) cells, including TF-expressing tumor cells. In some embodiments, the TF antibody binds to a TF extracellular epitope (e.g., an extracellular TF epitope) expressed on a cell (e.g., a tumor cell). In some embodiments, the TF is human TF. An exemplary amino acid sequence of human TF is described herein (SEQ ID NO: 175).

[0388] In some embodiments, a TF antibody competes for binding to TF with a reference TF antibody that comprises the VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 of any one of the antibodies described herein (e.g., the amino acid sequences of the VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 set forth in any one of Tables 1-2). Thus, in some embodiments, a TF antibody competes for binding to TF with a reference TF antibody that comprises one, two, and / or three VH CDRs and / or one, two, and / or three VL CDRs from (a) the antibody designated EXMA-006, or (b) the antibody designated EXMA-007. In some embodiments, a TF-ADC comprises a drug conjugated (directly or indirectly) to a TF antibody, where the TF antibody competes for binding to TF with a reference TF antibody comprising one, two, and / or three VH CDRs and one, two, and / or three VL CDRs from (a) the antibody designated EXMA-006 or (b) the antibody designated EXMA-007. In some embodiments, the TF antibody competes for binding to TF with a reference TF antibody comprising one, two, and / or three VH CDRs and one, two, and / or three VL CDRs from (a) the antibody designated EXMA-006 or (b) the antibody designated EXMA-007. In some embodiments, the TF antibody competes for binding to TF with a reference TF antibody comprising (a) a VH region comprising the amino acid sequence of SEQ ID NO:25 and a VL region comprising the amino acid sequence of SEQ ID NO:26, or (b) a VH region comprising the amino acid sequence of SEQ ID NO:41 and a VL region comprising the amino acid sequence of SEQ ID NO:42. In some embodiments, other suitable TF antibodies (see, e.g., International Publication Nos. WO2019136309 and WO2021003399) may be used, each of which is incorporated by reference herein in its entirety.

[0389] In some embodiments, the TF antibody comprises the VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 of any one of the antibodies described herein (e.g., the amino acid sequences of the VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 set forth in any one of Tables 1-2). Thus, in some embodiments, the TF antibody comprises one, two, and / or three heavy chain CDRs and / or one, two, and / or three light chain CDRs from (a) the antibody designated EXMA-006 or (b) the antibody designated EXMA-007. In some embodiments, the TF antibody comprises one, two, and / or three heavy chain CDRs and one, two, and / or three light chain CDRs from (a) the antibody designated EXMA-006 or (b) the antibody designated EXMA-007.

[0390] In some embodiments, the TF antibody comprises a VH region comprising one or more (e.g., one, two, or three) VH CDR1, VH CDR2, and VH CDR3 described herein (e.g., in any one of Tables 1-2), and / or a VL region comprising one or more (e.g., one, two, or three) VL CDR1, VL CDR2, and VL CDR3 described herein (e.g., in any one of Tables 1-2). In some embodiments, the TF antibody is bispecific, comprising a first binding region comprising one, two, and / or three heavy chain CDRs and / or one, two, and / or three light chain CDRs described in any one of Tables 1-2, and a second region comprising one, two, and / or three heavy chain CDRs and / or one, two, and / or three light chain CDRs from a binding agent that binds to a second target antigen that is not TF. In some embodiments, the TF antibody is bispecific and comprises a first binding domain comprising one, two, and / or three heavy chain CDRs and / or one, two, and / or three light chain CDRs as set forth in any one of Tables 1-2, and a second binding domain comprising one, two, and / or three heavy chain CDRs and / or one, two, and / or three light chain CDRs from a binding agent that binds to a second TF epitope.

[0391] The antibody designated EXMA-006 comprises the VH amino acid sequence of SEQ ID NO:25 and the VL amino acid sequence of SEQ ID NO:26.

[0392] The antibody designated EXMA-007 comprises the VH amino acid sequence of SEQ ID NO:41 and the VL amino acid sequence of SEQ ID NO:42. [Table 1] [Table 2]

[0393] In some embodiments, the TF antibody comprises a VH region. In some embodiments, the TF antibody comprises a VL region. In some embodiments, the TF antibody has a combination of (i) a VH region and (ii) a VL region.

[0394] In some embodiments, a TF antibody comprises a heavy chain having a combination of (i) a VH described herein (e.g., in any one of Tables 1-2) and (ii) one or more heavy chain constant regions (e.g., CH1, hinge, CH2, and CH3). An exemplary IgG heavy chain can comprise any VH amino acid sequence described herein and the following CH1, hinge, CH2, and CH3 amino acid sequences: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 89).

[0395] In some embodiments, a TF antibody comprises a heavy chain having a combination of (i) a VH described herein (e.g., in any one of Tables 1-2) and (ii) one or more heavy chain constant regions (e.g., CH1, hinge, CH2, and CH3). An exemplary IgG heavy chain can comprise any VH amino acid sequence described herein and the following CH1, hinge, CH2, and CH3 amino acid sequences: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 90).

[0396] In some embodiments, a TF antibody comprises a light chain having a combination of (i) a VL region described herein (e.g., in any one of Tables 1-2) and (ii) a light chain constant region (CL). An exemplary light chain (e.g., for pairing with an IgG heavy chain) can comprise any VL amino acid sequence described herein and the following CL amino acid sequence: RTVAAPSVFIFPPSDSQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 95).

[0397] In some embodiments, a TF antibody comprises a light chain having a combination of (i) a VL region described herein (e.g., in any one of Tables 1-2) and (ii) a light chain constant region (CL). An exemplary light chain (e.g., for pairing with an IgG heavy chain) can comprise any VL amino acid sequence described herein and the following CL amino acid sequence: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 96).

[0398] In some embodiments, the TF antibody comprises: (a) a heavy chain having a combination of (i) a VH described herein (e.g., in any one of Tables 1-2); and (ii) one or more heavy chain constant regions (e.g., CH1, hinge, CH2, and CH3); and (b) a light chain having a combination of (i) a VL described herein (e.g., in any one of Tables 1-2); and (ii) an IgG-format light chain constant region (CL1).

[0399] In some embodiments, the TF antibody comprises: a VH (having the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYTFDVYGISWVRQAPGQGLEWMGWIAPYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDAGTYSPFGYGMDVWGQGTTVTVSS (SEQ ID NO: 25)); and VL (having the following amino acid sequence: DIQMTQSPSTLSASVGDRVTITCQASQSINNWLAWYQQKPGKAPKLLIYKAYNLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQLFQSLPPFTFGGGTKVEIK (SEQ ID NO: 26)).

[0400] In some embodiments, the TF antibody comprises: a VH (having the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYTFDAYGISWVRQAPGQGLEWMGWIAPYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDAGTYSPFGYGMDVWGQGTTVTVSS (SEQ ID NO: 41)); and VL (having the following amino acid sequence: DIQMTQSPSTLSASVGDRVTITCRASESISNWLAWYQQKPGKAPKLLIYKAYSLEYGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQFQKLPPFTFGGGTKVEIK (SEQ ID NO: 42).

[0401] In some embodiments, the antibody that binds to TF has a heavy chain (having the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYTFDVYGISWVRQAPGQGLEWMGWIAPYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDAGTYSPFGYGMDVWGQGTTVT VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEL LGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 93)); and a light chain (having the following amino acid sequence: DIQMTQSPSTLSASVGDRVTITCQASQSINNWLAWYQQKPGKAPKLLIYKAYNLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQLFQSLPPFTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 91)).

[0402] In some embodiments, an antibody that binds to TF has a heavy chain (having the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYTFDAYGISWVRQAPGQGLEWMGWIAPYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDAGTYSPFGYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL and a light chain (having the following amino acid sequence: DIQMTQSPSTLSASVGDRVTITCRASESISNWLAWYQQKPGKAPKLLIYKAYSLEYGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQFQKLPPFTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 92)).

[0403] In some embodiments, one or both of the heavy chains of the TF antibody further comprises a signal peptide, e.g., at the N-terminus of the chain. Additionally or alternatively, one or both of the light chains of the TF antibody further comprises a signal peptide, e.g., at the N-terminus of the chain. In some embodiments, the signal peptide comprises the amino acid sequence of MMSFVSLLLVGILFHATQA (SEQ ID NO: 97). In some embodiments, the signal peptide comprises the amino acid sequence of MGWSLILLFLVAVATRVHS (SEQ ID NO: 98).

[0404] In some embodiments, the TF antibody comprises a heavy chain with a signal peptide and therefore comprises the following amino acid sequence: [ka]

[0405] In some embodiments, the TF antibody comprises a heavy chain with a signal peptide and therefore comprises the following amino acid sequence: [ka] (The underlined amino acids represent the signal peptide sequence).

[0406] In some embodiments, a TF antibody comprises one or more CDRs (e.g., 1, 2, 3, 4, 5, or 6 CDRs), e.g., a VH CDR1, a VH CDR2, a VH CDR3, a VL CDR1, a VL CDR2, and / or a VL CDR3, as set forth in Table 1. In some embodiments, a TF antibody comprises one or more CDRs (e.g., 1, 2, 3, 4, 5, or 6 CDRs), e.g., a VH CDR1, a VH CDR2, a VH CDR3, a VL CDR1, a VL CDR2, and / or a VL CDR3, as set forth in Table 2.

[0407] In some embodiments, the TF antibody comprises one or more CDRs (e.g., one, two, or three VH CDRs), e.g., VH CDR1, VH CDR2, VH CDR3, as set forth in Table 1. In some embodiments, the TF antibody comprises one or more CDRs (e.g., one, two, or three VL CDRs), e.g., VL CDR1, VL CDR2, and / or VL CDR3, as set forth in Table 1. In some embodiments, the TF antibody comprises one or more CDRs (e.g., one, two, or three VH CDRs), e.g., VH CDR1, VH CDR2, VH CDR3, as set forth in Table 1, and one or more CDRs (e.g., one, two, or three VL CDRs), e.g., VL CDR1, VL CDR2, and / or VL CDR3, as set forth in Table 1.

[0408] In some embodiments, the TF antibody comprises one or more CDRs (e.g., one, two, or three VH CDRs), e.g., VH CDR1, VH CDR2, VH CDR3, as set forth in Table 2. In other embodiments, the TF antibody comprises one or more CDRs (e.g., one, two, or three VL CDRs), e.g., VL CDR1, VL CDR2, and / or VL CDR3, as set forth in Table 2. In some embodiments, the TF antibody comprises one or more CDRs (e.g., one, two, or three VH CDRs), e.g., VH CDR1, VH CDR2, VH CDR3, as set forth in Table 2, and one or more CDRs (e.g., one, two, or three VL CDRs), e.g., VL CDR1, VL CDR2, and / or VL CDR3, as set forth in Table 2.

[0409] In some embodiments, a TF antibody comprises one or more (e.g., one, two, or three) VH CDRs described herein (e.g., in Tables 1-2). In other embodiments, a TF antibody comprises one or more (e.g., one, two, or three) VL CDRs described herein (e.g., in Tables 1-2). In some embodiments, a TF antibody comprises one or more (e.g., one, two, or three) VH CDRs described herein (e.g., in Tables 1-2) and one or more (e.g., one, two, or three) VL CDRs described herein (e.g., in Tables 1-2). Thus, in some embodiments, a TF antibody comprises a VH CDR1 comprising an amino acid sequence comprising any one of SEQ ID NOs: 1, 7, 8, 15, 21, 27, 31, 32, 35, and 39. In some embodiments, a TF antibody comprises a VH CDR2 comprising an amino acid sequence comprising any one of SEQ ID NOs: 2, 9, 14, 16, and 22. In some embodiments, the TF antibody comprises a VH CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 3, 10, 17, and 23. In some embodiments, the TF antibody comprises a VH CDR1 and / or a VH CDR2 and / or a VH CDR3 independently selected from the VH CDR1, VH CDR2, and VH CDR3 set forth in any one of Tables 1-2. In some embodiments, the TF antibody comprises a VL CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 4, 11, 18, 24, 28, 33, 36, and 40. In some embodiments, the TF antibody comprises a VL CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 5, 12, 19, 29, and 37. In some embodiments, the TF antibody comprises a VL CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 6, 13, 20, 30, 34, and 38. In some embodiments, the TF antibody comprises a VL CDR1 and / or a VL CDR2 and / or a VL CDR3 independently selected from a VL CDR1, VL CDR2, VL CDR3 described herein (e.g., any one of Tables 1-2).

[0410] In some embodiments, a TF-ADC comprises a TF antibody, wherein the antibody comprises all three heavy chain CDRs and / or all three light chain CDRs from: (a) an antibody comprising the VH amino acid sequence of SEQ ID NO: 25 and the VL amino acid sequence of SEQ ID NO: 26 (designated EXMA-006), or (b) an antibody comprising the VH amino acid sequence of SEQ ID NO: 41 and the VL amino acid sequence of SEQ ID NO: 42 (designated EXMA-007). In some embodiments, a TF-ADC comprises a TF antibody, wherein the TF antibody comprises all three heavy chain CDRs and / or all three light chain CDRs from the antibody designated EXMA-006. In some embodiments, a TF-ADC comprises a TF antibody, wherein the TF antibody comprises all three heavy chain CDRs and / or all three light chain CDRs from the antibody designated EXMA-006.

[0411] In some embodiments, a TF-ADC comprises a TF antibody, wherein the antibody comprises (a) a VH region comprising the VH CDR1, VH CDR2, and VH CDR3 amino acid sequences described herein (e.g., in Tables 1-2); and / or (b) a VL region comprising the VL CDR1, VL CDR2, and VL CDR3 amino acid sequences described herein (e.g., in Tables 1-2). In some embodiments, a TF-ADC comprises a TF antibody, wherein the antibody comprises a VH region comprising the VH CDR1, VH CDR2, and VH CDR3 amino acid sequences described herein (e.g., in Tables 1-2). In some embodiments, a TF-ADC comprises a TF antibody, wherein the antibody comprises a VL region comprising the VL CDR1, VL CDR2, and VL CDR3 amino acid sequences described herein (e.g., in Tables 1-2).

[0412] In some embodiments, the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

[0413] In some embodiments, the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO:7, a VH CDR2 comprising the amino acid sequence of SEQ ID NO:2, a VH CDR3 comprising the amino acid sequence of SEQ ID NO:3, a VL CDR1 comprising the amino acid sequence of SEQ ID NO:4, a VL CDR2 comprising the amino acid sequence of SEQ ID NO:5, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO:6.

[0414] In some embodiments, the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO:8, a VH CDR2 comprising the amino acid sequence of SEQ ID NO:9, a VH CDR3 comprising the amino acid sequence of SEQ ID NO:10, a VL CDR1 comprising the amino acid sequence of SEQ ID NO:11, a VL CDR2 comprising the amino acid sequence of SEQ ID NO:12, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO:13.

[0415] In some embodiments, the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 4, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

[0416] In some embodiments, the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 15, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 16, a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 17, a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 18, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 19, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 20.

[0417] In some embodiments, the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 21, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 23, a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 24, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 12, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 6.

[0418] In some embodiments, the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 27, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 28, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 29, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 30.

[0419] In some embodiments, the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 31, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 28, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 29, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 30.

[0420] In some embodiments, the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 32, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 9, a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 10, a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 33, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 12, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 34.

[0421] In some embodiments, the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 27, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 14, a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 28, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 29, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 30.

[0422] In some embodiments, the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 35, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 16, a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 17, a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 36, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 37, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 38.

[0423] In some embodiments, the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 39, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 22, a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 23, a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 40, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 12, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 30.

[0424] In some embodiments, a TF-ADC comprises a TF antibody, wherein the antibody comprises a VH region and / or a VL region described herein, wherein the VH and / or VL comprise human framework sequences. In some embodiments, the VH and / or VL region comprises framework 1 (FR1), framework 2 (FR2), framework 3 (FR3), and / or framework 4 (FR4) sequences, e.g., human FR1, human FR2, human FR3, and / or human FR4.

[0425] In some embodiments, the CDRs of TF antibodies can be determined using the Kabat system (Kabat et al. (1971) Ann. NY Acad. Sci. 190:382-391 and, Kabat et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USDapartment of Health and Human Services, NIH Publication No. 91-3242).

[0426] In some embodiments, the CDRs of a TF antibody can be determined using the Chothia system, which will be referred to herein as "Chothia CDRs" (see, e.g., Chothia and Lesk, 1987, J. Mol. Biol., 196:901-917; Al-Lazikani et al., 1997, J. Mol. Biol., 273:927-948; Chothia et al., 1992, J. Mol. Biol., 227:799-817; Tramontano A. et al., 1990, J. Mol. Biol. 215(1):175-82; and U.S. Patent No. 7,709,226).

[0427] In some embodiments, the CDRs of TF antibodies can be determined ("IMGT® CDRs") using the ImMunoGeneTics (IMGT®) system, e.g., as described in Lefranc, M.-P., 1999, The Immunologist, 7:132-136 and Lefranc, M.-P. et al., 1999, Nucleic Acids Res., 27:209-212.

[0428] In some embodiments, the CDRs of TF antibodies can be determined using the AbM system, which will be referred to herein as "AbM CDRs" (e.g., as described in MacCallum et al., 1996, J. Mol. Biol., 262:732-745). See also, e.g., Martin, A., "Protein Sequence and Structure Analysis of Antibody Variable Domains," in Antibody Engineering, Kontermann and Dubel, eds., Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001).

[0429] In some embodiments, the CDRs of TF antibodies can be determined using the Contact system, which will be referred to herein as "Contact CDRs" (see, e.g., MacCallum RM et al., 1996, J Mol Biol 5:732-745). Contact CDRs are based on analysis of available complex crystal structures.

[0430] In some embodiments, the TF antibody comprises: a heavy chain variable region (VH) ((1) a VH CDR1 comprising the amino acid sequence of any one of (i) SEQ ID NO:1, (ii) SEQ ID NO:7, (iii) SEQ ID NO:8, (iv) SEQ ID NO:15, and (v) SEQ ID NO:21; (2) a VH CDR2 comprising the amino acid sequence of any one of (i) SEQ ID NO:2, (ii) SEQ ID NO:9, (iii) SEQ ID NO:14, (iv) SEQ ID NO:16, and (v) SEQ ID NO:22; and (3) a VH CDR3 comprising the amino acid sequence of any one of (i) SEQ ID NO:3, (ii) SEQ ID NO:10, (iii) SEQ ID NO:17; and (iv) SEQ ID NO:23); and / or a VL region ((1) a VL CDR1 comprising the amino acid sequence of any one of (i) SEQ ID NO:4, (ii) SEQ ID NO:11, (iii) SEQ ID NO:18, and (iv) SEQ ID NO:24; (2) a VL CDR2 comprising the amino acid sequence of any one of (i) SEQ ID NO:5, (ii) SEQ ID NO:12, and (iii) SEQ ID NO:19). CDR2; and (3) a VL CDR3 comprising the amino acid sequence of any one of (i) SEQ ID NO: 6, (ii) SEQ ID NO: 13, and (iii) SEQ ID NO: 20).

[0431] In some embodiments, the TF antibody comprises the following: a...

Claims

1. Antibody-drug conjugates (ADCs) of formula (I) including the following: a. Antibodies that bind to tissue factor (TF), and b. Two or more drugs, each conjugated to a pyridazine-pyrrolo coupling site via a linker, 【Transformation 67】 (In the formula, Ab represents the antibody that binds to TF, Z １ Z ２ , and Z ４ However, each operates independently, CR ４ And, Z ３ However, C-L Ｂ -W ２ And, R １ , R ２ , R ３ , and R ４ are each selected from hydrogen and alkyl, L Ａ However, it is the first linker, which includes the following: -(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, 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. T １ , T ２ , T ３ , T ４ , T ５ , and T ６ However, each is independent, covalently bonded, (C １ -C １２ ) Alkylene, substitution (C １ -C １２ ) Alkylene, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA) ｗ (PEG) ｎ (AA) ｐ ,-(CR １３ OH) ｘ -, selected from 4-aminopiperidine (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 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 However, each is independent of covalent bonds, -CO-, and -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 Selected from the group consisting of - and -P(O)OH-, where each q is an integer from 1 to 6, Each R １３ However, they are independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. Each R １５ However, they are 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 However, it is a second linker that includes the following: -(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 -、 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 ) alkylene, substituted (C １ -C １２ ) alkylene, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA) ｗ , (PEG) ｎ , (AA) ｐ , -(CR １３ OH) ｘ -, 4-aminopiperidine (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), an acetal group, hydrazine, disulfide, and ester; wherein 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 of 1 to 20, each n is an integer of 1 to 30, each p is an integer of 1 to 20, and each x is an integer of 1 to 12, V ７ , V ８ , V ９ , V １０ , V １１ , V １２ , and V １３ However, each is independent of covalent bonds, -CO-, and -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 Selected from the group consisting of - and -P(O)OH-, where each q is an integer from 1 to 6, Each R １３ However, they are independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. Each R １５ However, they are 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 However, it is the first drug, W ２ However, it is the second drug.

2. T １ However, (C １ -C １２ ) Alkylene and substitution (C １ -C １２ ) Selected from alkylenes, T ２ , T ３ , T ４ , T ５ , and T ６ However, each is independent, covalently bonded, (C １ -C １２ ) Alkylene, substitution (C １ -C １２ ) Alkylene, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA) ｗ (PEG) ｎ (AA) ｐ ,-(CR １３ OH) ｘ - Selected from 4-aminopiperidine (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 However, each is independent of covalent bonds, -CO-, and -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 Selected from the group consisting of - and -P(O)OH-, (PEG) ｎ but, 【Transformation 68】 And, EDA is an ethylenediamine moiety having the following structure; 【Transformation 69】 4-aminopiperidine (4AP) 【Transformation 70】 And, Each R １２ However, independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, any two adjacent R １２ The groups are cyclically bonded to form a piperazinyl ring, q is an integer from 1 to 6, r is 0 or 1, The ADC according to claim 1, wherein y is an integer from 1 to 6.

3. T １ However, (C １ -C １２ ) is alkylene, V １ However, it is -CONH-, T ２ However, substitution (C １ -C １２ ) is alkylene, V ２ However, it is -CO-, T ３ However, (AA) ｐ V ３ However, it does not exist. T ４ However, it is PABC, and V ４ However, it does not exist. p is an integer between 1 and 10, a, b, c, and d are each 1, The ADC according to claim 1 or 2, wherein e and f are each 0.

4. T ７ However, it is a covalent bond, T ８ , T ９ , T １０ , T １１ , and T １２ However, each is independent, covalently bonded, (C １ -C １２ ) Alkylene, substitution (C １ -C １２ ) Alkylene, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA) ｗ (PEG) ｎ (AA) ｐ ,-(CR １３ OH) ｘ - Selected from 4-aminopiperidine (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 However, each is independent of covalent bonds, -CO-, and -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 Selected from the group consisting of - and -P(O)OH-, (PEG) ｎ but, 【Chemistry 71】 And n is an integer from 1 to 30. EDA is an ethylenediamine moiety having the following structure: 【Chemistry 72】 y is an integer from 1 to 6, and r is 0 or 1. 4-aminopiperidine (4AP) 【Transformation 73】 And, Each R １２ However, independently selected from hydrogen, alkyl, substituted alkyl, polyethylene glycol moiety, aryl, and substituted aryl, any two adjacent R １２ The groups are cyclically bonded to form a piperazinyl ring, g, h, i, j, and k are each 1. The ADC according to claim 1, wherein l and m are 0.

5. (i) T ７ However, it does not exist, V ７ However, it is NHKCO, T ８ However, (C １ -C １２ ) is alkylene, V ８ However, it is -CONH-, T ９ However, substitution (C １ -C １２ ) is alkylene, V ９ However, it is -CO-, T １０ However, (AA) ｐ V １０ However, it does not exist. T １１ However, it is PABC, and V １１ However, it does not exist. p is an integer between 1 and 10, g, h, i, j, and k are each 1. l and m are both 0, (ii) T1 is (C1-C12)alkylene, and V1 is -CONH-, T2 is a substituted (C1-C12) alkylene, and V2 is -CO-. T3 is (AA)p (where p is an integer from 1 to 20), and V3 is a covalent bond. T4 is PABC, and V4 is a covalent bond. a, b, c, and d are each 1, e and f are both 0, T7 is a covalent bond, and V7 is -NHCO-. T8 is (C1-C12)alkylene, and V8 is -CONH-, T9 is a substituted (C1-C12) alkylene, and V9 is -CO-. T 10 is (AA) p (where p is an integer from 1 to 20), and V 10 is a covalent bond. T 11 is PABC, and V 11 is a covalent bond. h, i, j, and k are each 1. l and m are either 0 or (iii) T1 is (C1-C12)alkylene, and V1 is -CONH-, T2 is an alkylene (C1-C6) substituted with -NHCO(PEG)k (where k is an integer from 2 to 10), and V2 is -CO-. T3 is (AA)2, and V3 is a covalent bond. T4 is a PABC substituted with a glycoside, and V4 is a covalent bond. a, b, c, and d are each 1, e and f are both 0, T7 is a covalent bond, and V7 is -NHCO-. T8 is (C1-C6) alkyl, and V8 is -CONH-, T9 is an alkylene (C1-C6) substituted with -NHCO(PEG)k (where k is an integer from 2 to 10), and V9 is -CO-. T 10 is (AA) 2, and V 10 is a covalent bond. T 11 is a PABC substituted with a glycoside, and V 11 is a covalent bond. h, i, j, and k are each 1. l and m are both 0. The ADC according to claim 1. Claim 6 (i) T ２ and T ９ one or both of are substituted with -NHCO(PEG) ｋ substituted (C １ to C ６ ) alkylene, and k is an integer of 2 to 10, (ii) p is 2, (iii)s is 2 or 4 (iv) One or both of W1 and W2 are camptothecin analogs. (v) The camptothecin analog is belotecan, and / or (vi) W1 and W2 are each Belotecan, The ADC according to claim 1.

7. ADC represented by equation (II): 【Chemistry 74】 (In the formula, Ab represents the antibody that binds to TF, (where s is an integer between 1 and 10, or s is 2 or 4).

8. Ab is, (A) (i) VH CDR1, VH CDR2, and VH CDR3 described in VH containing the amino acid sequence of SEQ ID NO: 25, and VL CDR1, VL CDR2, and VL CDR3 described in VL containing the amino acid sequence of SEQ ID NO: 26; or (ii) VH CDR1, VH CDR2, and VH CDR3 described in VH containing the amino acid sequence of SEQ ID NO: 41, and VL CDR1, VL CDR2, and VL CDR3 described in VL containing the amino acid sequence of SEQ ID NO: 42, and optionally the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 is determined according to the Kabat numbering rules, Chothia numbering rules, AbM numbering rules, Contact numbering rules, IMGT numbering rules, or a combination of Kabat and Chothia, and / or (B) (i) A VH region comprising VH CDR1 containing the amino acid sequence of SEQ ID NOs: 1, 7, 8, 15, 21, 27, 31, 32, 35, or 39; VH CDR2 containing the amino acid sequence of SEQ ID NOs: 2, 9, 14, 16, or 22; and VH CDR3 containing the amino acid sequence of SEQ ID NOs: 3, 10, 17, or 23; and a VL region comprising VL CDR1 containing the amino acid sequence of SEQ ID NOs: 4, 11, 18, 24, 28, 33, 36, or 40; VL CDR2 containing the amino acid sequence of SEQ ID NOs: 5, 12, 19, 29, or 37; and VL CDR3 containing the amino acid sequence of SEQ ID NOs: 6, 13, 20, 30, 34, or 38, The ADC according to claim 1 or 7.

9. Ab is, (i) One or more of the Framework 1 (FR1), Framework 2 (FR2), Framework 3 (FR3), and Framework 4 (FR4) sequences described in any one of Sequence IDs 25, 26, 41, and 42, or (ii) Human framework sequence The ADC according to claim 1 or 7, including the ADC described in claim 1 or 7.

10. Ab is, (i) containing VH containing the amino acid sequence of SEQ ID NO: 25 and VL containing the amino acid sequence of SEQ ID NO: 26, (ii) Contains a heavy chain containing the amino acid sequence of SEQ ID NO: 82, (iii) A heavy chain containing the amino acid sequence of SEQ ID NO: 82 and a light chain containing the amino acid sequence of SEQ ID NO: 91, (iv) Contains a heavy chain containing the amino acid sequence of SEQ ID NO: 79, (v) comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 79 and a light chain containing the amino acid sequence of SEQ ID NO: 91, (vi) containing VH containing the amino acid sequence of SEQ ID NO: 41 and VL containing the amino acid sequence of SEQ ID NO: 42, (vii) Contains a heavy chain containing the amino acid sequence of SEQ ID NO: 85, (viiii) A heavy chain containing the amino acid sequence of SEQ ID NO: 85 and a light chain containing the amino acid sequence of SEQ ID NO: 92, (ix) Contains a heavy chain containing the amino acid sequence of SEQ ID NO: 88, or (x) A heavy chain containing the amino acid sequence of SEQ ID NO: 88 and a light chain containing the amino acid sequence of SEQ ID NO: 92 The ADC according to claim 1 or 7.

11. A pharmaceutical composition comprising the ADC described in claim 1 or 7 and a pharmaceutically acceptable excipient.

12. The pharmaceutical composition according to claim 11, wherein the ADC drug-to-antibody ratio (DAR) is about 1 to about 20, or about 2 to about 8, or about 4, or about 8.

13. A method for treating a target cancer or tumor, comprising administering to the target an ADC according to claim 1 or 7, or a pharmaceutical composition comprising an ADC according to claim 1 or 7 and a pharmaceutically acceptable excipient.

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