Antibody-drug conjugates and methods of their use

Abstract

The present disclosure provides methods of treatment of a colorectal cancer (CRC) or a non-small cell lung cancer (NSCLC) comprising administering anti-tissue factor antibody-drug conjugate structures. The antibody-drug conjugate structures include a branched linker, where two or more payloads per branched linker are attached to an antibody. In addition, the disclosure also encompasses compounds and methods for production of such conjugates.

Description

Attorney Docket No. 14529-193-228ANTIBODY-DRUG CONJUGATES AND METHODS OF THEIR USE CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Patent Application No.63 / 775,951, filed March 21, 2025, the disclosure of which is incorporated by reference herein in its entirety.SEQUENCE LISTING

[0002] This application contains an electronic Sequence Listing which has been submitted in XML file format with this application, the entire content of which is incorporated by reference herein in its entirety. The Sequence Listing XML file submitted with this application is entitled “14529-193 -228_SEQ_L1 STING. xml”, was created on March 4, 2026, and is 193,600 bytes in size.FIELD

[0003] The present disclosure relates generally to methods of using antibody-drug conjugates (ADCs) that bind to tissue factor (TF, e.g., human TF) in treating a cancer selected from a colorectal and non-small cell lung cancer.INTRODUCTION

[0004] Blood coagulation involves a complex set of processes that result in blood clotting. Tissue factor (TF) plays an important role in these coagulation processes. TF is a cell surface receptor for the serine protease factor Vila (FVIIa). The TF / FVIIa complex catalyzes conversion of the inactive protease factor X (FX) into the active protease factor Xa (FXa). FXa and its co-factor FVa form the prothrombinase complex, which generates thrombin from prothrombin. Thrombin converts soluble fibrinogen into insoluble strands of fibrin and catalyzes many other coagulation-related processes. TF is over-expressed on multiple types of solid tumors. In cancer, TF / FVIIa signaling can support angiogenesis, tumor progression, and metastasis.

[0005] Antibody-drug conjugates (ADCs) have emerged over the past two decades as a new class of targeted-delivery therapies. A typical ADC includes an antibody -based targeting element attached to a highly potent pharmaceutical agent (payload) via a chemical linker using an available bioconjugation method. The molar ratio of targeting element (e.g., antibody) to attached payload can vary, and is referred to as the drug-to-antibody ratio (DAR). Commonly used bioconjugation methods either exploit endogenous amino acid residues of a protein ( / .< ., lysine and cysteine), or rely on selective engagement of a bioorthogonal functional group that has been intentionally introduced into the protein. As an1NAI-5011166969vlexample of the latter approach, the Hydrazino-z o-Pictet-Spengler (HIPS) conjugation method (FIG. 1) takes an advantage of an aldehyde functional group (an “aldehyde tag”), which can be introduced into a protein, such as an antibody, through various means (e.g., by the action of formyl generating enzyme (FGE)), serving as the conjugation handle. The aldehyde group cleanly reacts with the HIPS indole moiety to form a stable carbon-carbon bond that 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 alleviate TF-mediated diseases, disorders, or conditions, such as those involving tumor cells expressing TF.SUMMARY

[0007] Provided herein are methods for treating, preventing, or alleviating a cancer selected from colorectal cancer (CRC) and non-small cell lung cancer (NSCLC) in a subject. The methods comprise administering an ADC to the subject, wherein the ADC comprises an antibody that binds to tissue factor (“TF-ADC”). Such TF-ADCs, in some embodiments, bind to the same epitope of human TF as an antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL) described herein. In some embodiments, the colorectal cancer is adenocarcinoma. Additionally or alternatively, the colorectal cancer is metastatic to lung. In some embodiments, the non-small cell lung cancer is adenocarcinoma, squamous cell carcinoma, or large cell carcinoma.

[0008] In certain embodiments, the methods comprise administering pharmaceutical compositions comprising a TF-ADC that comprises an antibody or fragment thereof that binds to TF (“TF antibody”) and a drug conjugated (directly or indirectly) thereto. Such pharmaceutical compositions, in some embodiments, include TF-ADCs comprising an antibody or fragment thereof that binds to essentially the same epitope of human TF as an antibody comprising a VH and a VL described herein.

[0009] The present disclosure also provides methods of treating, preventing, or alleviating a TF-mediated colorectal cancer (CRC) and non-small cell lung cancer (NSCLC), such as alleviating one or more symptoms of the TF-mediated colorectal cancer (CRC) and non-small cell lung cancer (NSCLC) with a TF-ADC. In some embodiments, the colorectal cancer is adenocarcinoma. Additionally or alternatively, the colorectal cancer is metastatic to lung. In some embodiments, the non-small cell lung cancer is adenocarcinoma, squamous cell carcinoma, or large cell carcinoma.

[0010] More specifically, the present disclosure provides a method of treating, preventing, or alleviating a cancer selected from colorectal cancer (CRC) and non-small cell lung cancer 2NAI-5011166969vl(NSCLC) comprising administering a TF-ADC, wherein the TF-ADC comprises (a) a TF antibody and (b) one or more pyridazine-pyrrolo coupling moieties comprising a drug conjugated to the pyridazine-pyrrolo coupling moiety through a linker, for example, using the Hydrazino- / .w-Pictet-Spengler (HIPS) conjugation method. In some embodiments, the colorectal cancer is adenocarcinoma. Additionally or alternatively, the colorectal cancer is metastatic to lung. In some embodiments, the non-small cell lung cancer is adenocarcinoma, squamous cell carcinoma, or large cell carcinoma.

[0011] Traditionally, the HIPS conjugation method has been used to produce conjugates carrying one payload per HIPS moiety per aldehyde tag, which produces antibody conjugates with DAR values of up to 4. In some embodiments, a TF-ADC as disclosed herein comprises branched HIPS linkers that carry two (or more) molecules of the same or different payload per one HIPS moiety and are therefore capable of conjugating two (or more) small molecule payloads per one aldehyde group in a protein in a single conjugation step (FIG. 2).Consequently, the usage of such branched linkers allows the generation of higher DAR sitespecific conjugates (e.g., DAR up to 8) with controlled payload placement, which in the context of therapeutic ADCs would result in larger quantities of pharmaceutical agent delivered to the targeted tissue.

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

[0013] Aspects of the present disclosure include a TF-ADC comprising (a) a TF antibody; and (b) one or more pyridazine-pyrrolo coupling moieties comprising one or more drugs conjugated to the pyridazine-pyrrolo coupling moiety via one or more linkers. In one embodiment, TF-ADC is as disclosed, e.g, in International Patent Publication No. WO WO2024 / 054821 A2 (International Patent Application No. PCT / US2023 / 073516), incorporated herein by reference in its entirety.

[0014] In some embodiments of the methods provided herein, the TF-ADC is represented by Formula (I), the TF-ADC comprising:a. an antibody that binds to tissue factor (TF); andb. two or more drugs conjugated to a pyridazine-pyrrolo coupling moiety, each via a linker3NAI-5011166969vlwherein:Ab represents the antibody that binds to TF;Z1, Z2, and Z4are each independently CR4;Z3is C-LB-W2;R1, R2, R3, and R4are each selected from hydrogen and alkyl;LAis a first linker comprising:-(T1-V1)a-(T2-V2)b-(T3-V3)c-(T4-V4)d-(T5-V5)e-(T6-V6)f-, wherein:a, b, c, d, e and f are each independently 0 or 1, provided at least one of a, b, c, d, e, and f are 1 ;T1, T2, T3, T4, T5and T6are each independently selected from a covalent bond, (Ci-Cn)alkyl, substituted (Ci-Ci2)alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA)W, (PEG)n, (AA)P, -(CR13OH)X-, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxy-phenyl (PHP), an acetal group, a hydrazine, a disulfide, and an ester, wherein EDA is an ethylene diamine moiety, PEG is a polyethylene glycol, and AA is an amino acid residue or an amino acid analog, wherein 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;V1, V2, V3, V4,V5and V6are each independently selected from the group consisting of a covalent bond, -CO-, -NR15-, -NR15(CH2)q-, -NR15(C6H4)-, -CONR15-, -NR15CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR15-, -NR15SO2- and -P(O)OH-, wherein each q is an integer from 1 to 6;4NAI-5011166969vleach R13is independently selected from hydrogen, an alkyl, a substituted alkyl, an aryl, and a substituted aryl; andeach R15is 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;LBis a second linker comprising:-(T7-V7)g-(T8-V8)h-(T9-V9)i-(T10-V10)j-(T11-V11)k-(T12-V12)i-(T13-V13)m-, wherein: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;T7, T8, T9, T10, T11, T12and T13are each independently selected from a covalent bond, (Ci-Ci2)alkyl, substituted (Ci-Ci2)alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA)w, (PEG)n, (AA)P, -(CR13OH)X-, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxy-phenyl (PHP), an acetal group, a hydrazine, a disulfide, and an ester, wherein EDA is an ethylene diamine moiety, PEG is a polyethylene glycol, and AA is an amino acid residue or an amino acid analog, wherein 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;V7, V8, V9, V10,Vn, V12and V13are each independently selected from the group consisting of a covalent bond, -CO-, -NR15-, -NR15(CH2)q-, -NR15(CeH4)-, -CONR15-, -NR15CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR15-, -NR15SO2- and -P(O)OH-, wherein each q is an integer from 1 to 6;each R13is independently selected from hydrogen, an alkyl, a substituted alkyl, an aryl, and a substituted aryl; andeach R15is 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;5NAI-5011166969vlW1is a first drug; andW2is a second drug.

[0015] In some embodiments, s is 2. In some embodiments, s is 4.

[0016] In some embodiments, one or both of W1and W2are camptothecin analogues, for example, belotecan.

[0017] In some embodiments, LAcomprises:-(T1-V1)a-(T2-V2)b-(T3-V3)c-(T4-V4)d-(T5-V5)e-(T6-V6)f-, whereina, b, c, d, e and f are each independently 0 or 1, provided at least one of a, b, c, d, e, and f is 1;T1, T2, T3, T4, T5and T6are each independently selected from a covalent bond, (Ci-Cn)alkyl, substituted (Ci-Ci2)alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA)W, (PEG)n, (AA)P, -(CR13OH)X-, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxy-phenyl (PHP), an acetal group, a hydrazine, a disulfide, and an ester, wherein EDA is an ethylene diamine moiety, PEG is a polyethylene glycol, and AA is an amino acid residue or an amino acid analog, wherein 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;V1, V2, V3, V4,V5and V6are each independently selected from the group consisting of a covalent bond, -CO-, -NR15-, -NR15(CH2)q-, -NR15(C6H4)-, -CONR15-, -NR15CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR15-, -NR15SO2- and -P(O)OH-, wherein each q is an integer from 1 to 6;each R13is independently selected from hydrogen, an alkyl, a substituted alkyl, an aryl, and a substituted aryl; andeach R15is 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.

[0018] In some embodiments of LA:T1is selected from a (Ci-Ci2)alkyl and a substituted (Ci-Ci2)alkyl;6NAI-5011166969vlT2, T3, T4, T5and T6are each independently selected from a covalent bond, (Ci-Cn)alkyl, substituted (Ci-Ci2)alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA)W, (PEG)n, (AA)p, -(CR13OH)X-, 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PAB A, PAP, PHP, an acetal group, a hydrazine, and an ester; andV1, V2, V3, V4,V5and V6are each independently selected from the group consisting of a covalent bond, -CO-, -NR15-, -NR15(CH2)q-, -NR15(C6H4)-, -CONR15-, -NR15CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2- , -SO2NR15-, -NR15SO2-, and -P(O)OH-; wherein:EDA is an ethylene diamine moiety having the following structure:, where y is an integer from 1 to 6 and r is 0 or 1;4-amino-piperidine (4AP) is ; andeach R12is independently selected from hydrogen, an alkyl, a substituted alkyl, a polyethylene glycol moiety, an aryl and a substituted aryl, wherein any two adjacent R12groups may be cyclically linked to form a piperazinyl ring.

[0019] In further embodiments, a, b, c, and d are each 1; and e and f are 0.

[0020] In some embodiments, T1, T2, T3, T4, T5and T6are each optionally substituted with a glycoside.

[0021] In some embodiments, MABO, MABC, PABO, PABC, PAB, PAB A, 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, LAis a linker wherein:T1is (Ci-Ci2)alkyl and V1is -CONH-;T2is substituted (Ci-Ci2)alkyl and V2is -CO-;T3is (AA)Pand V3is absent;T4is PABC and V4is absent;p is an integer from 1 to 10;7NAI-5011166969vla, b, c, and d are each 1; ande and f are each 0.

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

[0025] In some embodiments, LBcomprises:whereing, h, i, j, k, 1 and m are each independently 0 or 1, provided at least one of g, h, i, j, k, 1 and m is 1;T7, T8, T9, T10, T11, T12and T13are each independently selected from a covalent bond, (Ci-Ci2)alkyl, substituted (Ci-Ci2)alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA)w, (PEG)n, (AA)P, -(CR13OH)X-, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxy-phenyl (PHP), an acetal group, a hydrazine, a disulfide, and an ester, wherein EDA is an ethylene diamine moiety, PEG is a polyethylene glycol, and AA is an amino acid residue or an amino acid analog, wherein 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;V7, V8, V9, V10,Vn, V12and V13are each independently selected from the group consisting of a covalent bond, -CO-, -NR15-, -NR15(CH2)q-, -NR15(CeH4)-, -CONR15-, -NR15CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR15-, -NR15SO2- and -P(O)OH-, wherein each q is an integer from 1 to 6;each R13is independently selected from hydrogen, an alkyl, a substituted alkyl, an aryl, and a substituted aryl; andeach R15is 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, T7, T8, T9, T10, T11, T12and T13are each optionally substituted with a glycoside.8NAI-5011166969vl

[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] In some embodiments of LB:T7is a covalent bond;T8, T9, T10, T11and T12are each independently selected from a covalent bond, (Ci-Cnjalkyl, substituted (Ci-Cnjalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA)W, (PEG)n, (AA)p, -(CR13OH)X-, 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, an acetal group, a hydrazine, and an ester; andV7, V8, V9, V10,Vnand V12are each independently selected from the group consisting of a covalent bond, -CO-, -NR15-, -NR15(CH2)q-, -NR15(CeH4)-, -CONR15-, -NR15CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2- , -SO2NR15-, -NR15SO2-, and -P(O)OH-;wherein:(PEG)n is , where n is an integer from 1 to 30;EDA is an ethylene diamine moiety having the following structure:N \R12, where y is an integer from 1 to 6 and r is 0 or 1;N4-amino-piperidine (4AP) is R ; andeach R12is independently selected from hydrogen, an alkyl, a substituted alkyl, a polyethylene glycol moiety, an aryl and a substituted aryl, wherein any two adjacent R12groups may be cyclically linked to form a piperazinyl ring;g, h, i, j, and k are each 1; and1 and m is 0.

[0030] In some embodiments, T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, and T12are 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.9NAI-5011166969vl

[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, LBis a linker wherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CONH-;T9is substituted (Ci-Ci2)alkyl and V9is -CO-;T10is (AA)Pand V10is absent;T11is PABC and V11is absent;p is an integer from 1 to 10;g, h, i, j, and k are each 1; and1 and m are each 0.

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

[0035] In some embodiments, a TF-ADC is represented by Formula (I):(I)wherein:Ab represents the antibody that binds to TF;Z1, Z2, and Z4are each independently CR4;Z3is C-LB-W2;R1, R2, R3and R4are each selected from hydrogen and (Ci-Ci2)alkyl;LAis a first linker wherein:T1is (Ci-Ci2)alkyl and V1is -CONH-;T2is substituted (Ci-Ci2)alkyl and V2is -CO-;T3is (AA)Pwhere p is an integer from 1-20 and V3is a covalent bond;T4is PABC and V4is a covalent bond;a, b, c, and d are each 1;e and f are each 0; and10NAI-5011166969vlLBis a second linker whereinT7is a covalent bond and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CONH-;T9is substituted (Ci-Ci2)alkyl and V9is -CO-;T10is (AA)Pwhere p is an integer from 1-20 and V10is a covalent bond;T11is PABC and V11is a covalent bond; andg, h, i, j, and k are each 1; and1 and m are each 0;s is an integer from 1 to 10;W1is a first drug; andW2is a second drug.

[0036] In some embodiments, one or both of W1and W2are camptothecin analogues, for example, belotecan.

[0037] In some embodiments, a TF-ADC is represented by Formula (I):wherein:Ab represents the antibody that binds to TF;Z1, Z2, and Z4are each independently CR4;Z3is C-LB-W2;R1, R2, R3and R4are each selected from hydrogen and (Ci-Ci2)alkyl;LAis a linker wherein:T1is (Ci-Ce)alkyl and V1is -CONH-;T2is (Ci-Ce)alkylene substituted with -NHCO(PEG)k’, wherein k’ is an integer from 2 to 10 and V2is -CO-;T3is (AA)2and V3is a covalent bond;T4is PABC substituted with a glycoside and V4is a covalent bond;a, b, c, and d are each 1; ande and f are each 0; and11NAI-5011166969vlLBis a linker whereinT7is a covalent bond and V7is -NHCO-;T8is (Ci-Ce)alkyl and V8is -CONH-;T9is (Ci-Ce)alkylene substituted with -NHCO(PEG)k’, wherein k’ is an integer from 2 to 10 and V9is -CO-;T10is (AA)2 and V10is a covalent bond;T11is PABC substituted with a glycoside and V11is a covalent bond;g, h, i, j, and k are each 1; and1 and m are each 0;s is an integer from 1 to 10;W1is a first drug; andW2is a second drug.

[0038] In some embodiments, s is 2. In some embodiments, s is 4.

[0039] In some embodiments, the PABC of one or both of T4and T11is substituted with a glucuronide. In some embodiments, one or both of T1and T8is ethyl. In some embodiments, one or both of T2and T9is Cs alkylene substituted with -NHCO(PEG)k’, where k’ is an integer from 5-10. In some embodiments, one or both of W1and W2are camptothecin analogues, for example, belotecan.

[0040] In some embodiments, a TF-ADC is represented by Formula (II):12NAI-5011166969vlwherein:Ab represents the antibody that binds to TF; ands is an integer from 1 to 10.

[0041] In some embodiments, s is an integer from 1 to 4. In some embodiments, s is 2. In some embodiments, s is 4.

[0042] Formula (II) may be prepared by conjugating one or more linker-payloads of Formula (Ila), shown below, with a TF antibody:13NAI-5011166969vl

[0043] In some embodiments, a TF-ADC is produced by conjugating a linker-payload of Formula (Ila), shown above, with a TF antibody (Ab). In further embodiments, a TF-ADC is prepared by conjugating two linker-payloads of Formula (Ila), shown above, with a TF antibody (Ab). Accordingly, the TF-ADC has a drug-to-antibody ratio (DAR) of 4. In some embodiments, a TF-ADC is prepared by conjugating four linker-payloads of Formula (Ila), shown above, with a TF antibody (Ab). Accordingly, the TF-ADC has a drug-to-antibody ratio (DAR) of 8.

[0044] In some embodiments, a TF-ADC can be represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab), wherein Ab comprises: a VH CDR1, a VH CDR2, and a VH CDR3 as 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 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:26.

[0045] In some embodiments, a TF-ADC can be represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab), wherein Ab comprises: a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO:41 and a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:42.

[0046] In some embodiments, a TF-ADC can be represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab), wherein Ab comprises: (i) a 14NAI-5011166969vlVH region comprising a VH CDR1 comprising the amino acid sequence of SEQ ID NOs: 1, 7, 8, 15, 21, 27, 31, 32, 35, or 39, a VH CDR2 comprising the amino acid sequence of SEQ ID NOs:2, 9, 14, 16, or 22, and a VH CDR3 comprising the amino acid sequence of SEQ ID NOs:3, 10, 17, or 23; and (ii) a VL region comprising a VL CDR1 comprising the amino acid sequence of SEQ ID NOs:4, 11, 18, 24, 28, 33, 36, or 40, a VL CDR2 comprising the amino acid sequence of SEQ ID NOs:5, 12, 19, 29, or 37, and a VL CDR3 comprising the amino acid sequence of SEQ ID NOs:6, 13, 20, 30, 34, or 38.

[0047] In some embodiments, a TF-ADC can be represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab), wherein the Ab competes with any one of the TF antibodies as disclosed herein in binding to TF.

[0048] In some embodiments, a TF-ADC can be represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab), wherein Ab comprises a framework 1 (FR1), a framework 2 (FR2), a framework 3 (FR3) and / or a framework 4 (FR4) sequence as set forth in any one of SEQ ID NOs:25, 26, 41, and 42.

[0049] In some embodiments, a TF-ADC can be represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab), wherein Ab comprises human framework sequences.

[0050] In some embodiments, a TF-ADC can be represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab), wherein 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.

[0051] In some embodiments, a TF-ADC can be represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab), wherein 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) or produced by conjugating Formula (Ila) with a TF antibody (Ab), wherein 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.

[0052] In some embodiments, a TF-ADC can be represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab), wherein 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) or produced by 15NAI-5011166969vlconjugating Formula (Ila) with a TF antibody (Ab), wherein 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.

[0053] In some embodiments, a TF-ADC can be represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab), wherein Ab comprises 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.

[0054] In some embodiments, a TF-ADC can be represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab), wherein 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) or produced by conjugating Formula (Ila) with a TF antibody (Ab), wherein 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.

[0055] In some embodiments, a TF-ADC can be represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab), wherein 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) or produced by conjugating Formula (Ila) with a TF antibody (Ab), wherein 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.

[0056] The present disclosure also provides a pharmaceutical composition comprising (i) a TF-ADC, wherein the TF-ADC can be represented by Formula (I) or Formula (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab) and (ii) a pharmaceutically acceptable excipient, wherein the TF antibody (TF Ab or Ab) is as described in any embodiment described herein. In some embodiments, such a pharmaceutical composition can have a drug-to-antibody ratio (DAR) of the TF-ADC of about 1 to about 20, for example, a DAR of 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 8.16NAI-5011166969vl

[0057] In one embodiment, provided herein is a method for treating a cancer selected from a colorectal cancer and a non-small cell lung cancer in a subject comprising administering to the subject (i) the TF-ADC, wherein the TF-ADC can be represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab), or (ii) the pharmaceutical composition comprising a TF-ADC of Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab) and a pharmaceutically acceptable excipient, wherein the TF antibody is as described in any embodiment herein. In one embodiment, provided herein is a method for treating a colorectal cancer in a subject comprising administering to the subject (i) the TF-ADC, wherein the TF-ADC can be represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab), or (ii) the pharmaceutical composition comprising a TF-ADC of Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab) and a pharmaceutically acceptable excipient, wherein the TF antibody is as described in any embodiment herein. In one embodiment, provided herein is a method for treating a non-small cell lung cancer in a subject comprising administering to the subject (i) the TF-ADC, wherein the TF-ADC can be represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab) or (ii) the pharmaceutical composition comprising a TF-ADC of Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab) and a pharmaceutically acceptable excipient, wherein the TF antibody is as described in any embodiment herein. In some embodiments, the colorectal cancer is adenocarcinoma. Additionally or alternatively, the colorectal cancer is metastatic to lung. In some embodiments, the non-small cell lung cancer is adenocarcinoma, squamous cell carcinoma, or large cell carcinoma.

[0058] In one embodiment, the subject is human.BRIEF DESCRIPTION OF THE DRAWINGS

[0059] FIG. 1 shows a schematic drawing of the HIPS ligation for the synthesis of ADCs. Antibodies carrying aldehyde moieties are reacted with a Hydrazino-Ao-Pictet-Spengler (HIPS) linker and payload to generate a site-specifically conjugated ADC with a stable azacarboline linkage.

[0060] FIG. 2 shows a schematic representation of branched HIPS ligation for the synthesis of ADCs. Antibodies carrying four aldehyde moieties are reacted with a branched HIPS linker to generate ADCs with drug-to-antibody (DAR) value of up to 8, according to embodiments of the present disclosure.17NAI-5011166969vl

[0061] FIG. 3 provides a schematic drawing of TF-ADC 6-8. TF-ADC 6-8 utilizes SMARTAG® technology to conjugate a topoisomerase inhibitor (TOPOi) payload to a TF-targeting monoclonal antibody (mAb). The corresponding drug-antibody ratio (DAR) is 8.

[0062] FIGs. 4A-4C show in vitro internalization (FIG. 4A) and intracellular translocation (FIGs. 4B-4C) of TF-ADC 6-8 and its parental mAb. FIG. 4A shows the internalization of TF-ADC 6-8 and its parental mAb in A431 cells. Fluorescence-activated cell sorting (FACS) was used to measure the decrease of membrane-bound TF-ADC 6-8 or its parental mAb over time after initial binding. FIG. 4B provides confocal microscopy images showing TF-ADC 6-8 internalized to lysosomes. A431 cells were incubated with TF-ADC 6-8 for 24 hours. Scale bar, 10 pm. FIG. 4C provides quantification of TF-ADC 6-8 in lysosomal-associated membrane protein 1 (LAMP1) or early endosomal antigen 1 (EEA1) compartments using Pearson’s correlation coefficient (n = 10 images containing multiple cells). All data are represented as mean ± SEM. ****, p < 0.0001, One-way ANOVA. ANOVA, analysis of variance; SEM, standard error of the mean.

[0063] FIG. 5 shows that TF-ADC 6-8 demonstrated bystander tumor cell-killing activity. Mono-culture: Jurkat cells only; Co-culture: Jurkat cells plus MDA-MB-231 cells.

[0064] FIGs. 6A-6C show that TF-ADC 6-8 treatment induced immunogenic cell death assessed by high mobility group protein Bl (HMGB1) release (FIG. 6A), myeloid activation (FIG. 6B), and cytokine release (FIG. 6C). IFNy, interferon y; IP- 10, interferon-y induced protein 10; MMAE, monomethyl auristatin E; MIPla, macrophage inflammatory protein la; anti-PD-1, anti-PD-1 antibody (CAS Registry Number: 1374853-91-4); and TNFa, tumor necrosis factor alpha.

[0065] FIGs. 7A-7D show TF-ADC 6-8 mediated in vitro cytotoxicity and in vivo antitumor activity in TF-expressing cancer cell lines. FIG. 7A plots representative results of in vitro cytotoxicity tested using HPAF-II cells (TF antigen 142,998). FIG. 7B plots representative results of in vitro cytotoxicity tested using BxPC3 cells (TF antigen 142,952).FIG. 7C plots representative results of in vivo anti-tumor activity tested using HPAF-II xenograft. FIG. 7D plots representative results of in vivo anti-tumor activity tested using BxPC3 xenograft.

[0066] FIGs. 8A-8H show TF-ADC 6-8 mediated tumor volume regression and durable efficacy in CRC (FIGs. 8A-8C) and NSCLC (FIGs. 8D-8H) PDX models. FIGs. 8A-8C plots representative results obtained using three CRC PDX models (FIG. 8A, CRC-A-2 having a low H-score of 53; FIG. 8B, CRC-A-3 having an H-score of 146; and FIG. 8C, CRC-A-5 having a high H-score of 227). FIGs. 8D-8H plots representative results obtained 18NAI-5011166969vlusing five NSCLC PDX models (FIG. 8D, NSCLC-S-3 having an H-score of 187; FIG. 8E, NSCLC-L-1 having an H-score of 244; FIG. 8F, NSCLC-S-4 having an H-score of 122; FIG. 8G, NSCLC-S-2 having an H-score of 70; and FIG. 8H, NSCLC-S-1 having an H-score of 74). For all models, mean tumor volume (TV) was graphed as long as 80% animals remaining in treatment group. Arrows indicate dosing. *, P < 0.05; **, P < 0.01; ***, P < 0.001, one-way ANOVA, Tukey’s post-test or Kruskall-Wallis, Dunn’s post-test, ns, not significant. H-score, histology score.DETAILED DESCRIPTION

[0067] In one aspect, provided herein is a method for treating a cancer selected from a colorectal cancer (CRC) and a non-small cell lung cancer (NSCLC) in a subject, the method comprising administering an ADC to the subject at a therapeutically effective dose, wherein the ADC is represented by Formula (I) or (II) or produced by conjugating Formula (Ila) with a TF antibody (Ab). In some embodiments, the colorectal cancer is adenocarcinoma.Additionally or alternatively, the colorectal cancer is metastatic to lung. In some embodiments, the non-small cell lung cancer is adenocarcinoma, squamous cell carcinoma, or large cell carcinoma.DEFINITIONS

[0068] The following terms have the following meanings unless otherwise indicated. Any undefined terms have their art-recognized meanings.

[0069] “Alkyl” refers to monovalent saturated aliphatic hydrocarbyl groups having from 1 to 10 carbon atoms and such as 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, linear and branched hydrocarbyl groups such as 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-).

[0070] The term “substituted alkyl” refers to an alkyl group as defined herein wherein one or more carbon atoms in the alkyl chain (except the Ci carbon atom) have been optionally replaced with a heteroatom such as -O-, -N-, -S-, -S(O)n- (where n is 0 to 2), -NR- (where R is hydrogen or alkyl) and having from 1 to 5 substituents selected from the group consisting of 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,19NAI-5011166969vlaryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-aryl, -SO-heteroaryl, -SCh-alkyl, -SCh-aryl, -SO2-heteroaryl, and -NRaRb, wherein R and R may be the same or different and are chosen from hydrogen, optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic.

[0071] “Alkylene” refers to divalent aliphatic hydrocarbyl groups preferably having from 1 to 6 and more preferably 1 to 3 carbon atoms that are either straight-chained or branched, and which are optionally interrupted with one or more groups selected from -O-,-NR10-, -NR10C(O)-, -C(O)NR10- and the like. This term includes, by way of example, methylene (-CH2-), ethylene (-CH2CH2-), n-propylene (-CH2CH2CH2-), iso-propylene (-CH2CH(CH3)-), (-C(CH3)2CH2CH2-), (-C(CH3)2CH2C(O)-), (-C(CH3)2CH2C(O)NH-), (-CH(CH3)CH2-), and the like.

[0072] “ Substituted alkylene” refers to an alkylene group having from 1 to 3 hydrogens replaced with substituents as described for carbons in the definition of “substituted” below.

[0073] The term “alkane” refers to alkyl group and alkylene group, as defined herein.

[0074] The term “alkylaminoalkyl”, “alkylaminoalkenyl” and “alkylaminoalkynyl” refers to the groups R’NHR”- where R’ is alkyl group as defined herein and R” is alkylene, alkenylene or alkynylene group as defined herein.

[0075] The term “alkaryl” or “aralkyl” refers to the groups -alkylene-aryl and -substituted alkylene-aryl where alkylene, substituted alkylene and aryl are defined herein.

[0076] “Alkoxy” refers to the group -O-alkyl, wherein 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 as defined herein.

[0077] 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.

[0078] The term “alkoxyamino” refers to the group -NH-alkoxy, wherein alkoxy is defined herein.

[0079] The term “haloalkoxy” refers to the groups alkyl-O- wherein one or more hydrogen atoms on the alkyl group have been substituted with a halo group and include, by way of examples, groups such as trifluoromethoxy, and the like.20NAI-5011166969vl

[0080] The term “haloalkyl” refers to a substituted alkyl group as described above, wherein one or more hydrogen atoms on the alkyl group have been substituted with a halo group. Examples of such groups include, without limitation, fluoroalkyl groups, such as trifluoromethyl, difluoromethyl, trifluoroethyl and the like.

[0081] The term “alkylalkoxy” refers to the groups -alkylene-O-alkyl, alkylene-O-substituted alkyl, substituted alkylene-O-alkyl, and substituted alkylene-O-substituted alkyl wherein alkyl, substituted alkyl, alkylene and substituted alkylene are as defined herein.

[0082] The term “alkylthioalkoxy” refers to the group -alkylene-S-alkyl, alkylene-S-substituted alkyl, substituted alkylene-S-alkyl and substituted alkylene-S-substituted alkyl wherein alkyl, substituted alkyl, alkylene and substituted alkylene are as defined herein.

[0083] “Alkenyl” refers to straight chain or branched hydrocarbyl groups having from 2 to 6 carbon atoms and preferably 2 to 4 carbon atoms and having at least 1 and preferably from 1 to 2 sites of double bond unsaturation. This term includes, by way of example, bi-vinyl, allyl, and but-3-en-l-yl. Included within this term are the cis and trans isomers or mixtures of these isomers.

[0084] The term “substituted alkenyl” refers to an alkenyl group as defined herein having from 1 to 5 substituents, or from 1 to 3 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, -SCh-alkyl, -SCh-substituted alkyl, -SCh-aryl and -SCh-heteroaryl.

[0085] “Alkynyl” refers to straight or branched monovalent hydrocarbyl groups having from 2 to 6 carbon atoms and preferably 2 to 3 carbon atoms and having at least 1 and preferably from 1 to 2 sites of triple bond unsaturation. Examples of such alkynyl groups include acetylenyl (-C=CH), and propargyl (-CH2OCH).

[0086] The term “substituted alkynyl” refers to an alkynyl group as defined herein having from 1 to 5 substituents, or from 1 to 3 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,21NAI-5011166969vlheteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SCh-alkyl, -SCh-substituted alkyl, -SO2-aryl, and -SCh-heteroaryl.

[0087] “Alkynyloxy” refers to the group -O-alkynyl, wherein alkynyl is as defined herein. Alkynyloxy includes, by way of example, ethynyloxy, propynyloxy, and the like.

[0088] “Acyl” refers to the groups H-C(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)-, 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 are as defined herein. For example, acyl includes the “acetyl” group CH3C(O)-

[0089] “Acylamino” refers to the groups -NR20C(O)alkyl, -NR20C(O)substituted alkyl, N R20C(O)cycloalkyl, -NR20C(O) substituted cycloalkyl, - NR20C(O)cycloalkenyl, -NR20C(O)substituted cycloalkenyl, -NR20C(O)alkenyl, -NR20C(O)substituted alkenyl, -NR20C(O)alkynyl, -NR20C(O) substitutedalkynyl, -NR20C(O)aryl, -NR20C(O)substituted aryl, -NR20C(O)heteroaryl, -NR20C(O)substituted heteroaryl, -NR20C(O)heterocyclic, and -NR20C(O)substituted heterocyclic, wherein R20is hydrogen or alkyl and 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 are as defined herein.

[0090] “Aminocarbonyl” or the term “aminoacyl” refers to the group -C(O)NR21R22, wherein R21and R22independently are 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 and where R21and R22are optionally joined together with the nitrogen bound thereto to form a heterocyclic or substituted heterocyclic group, and 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 are as defined herein.22NAI-5011166969vl

[0091] “Aminocarbonylamino” refers to the group -NR21C(O)NR22R23where R21, R22, and R23are independently selected from hydrogen, alkyl, aryl or cycloalkyl, or where two R groups are joined to form a heterocyclyl group.

[0092] The term “alkoxycarbonylamino” refers to the group -NRC(O)OR where each R is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, or heterocyclyl wherein alkyl, substituted alkyl, aryl, heteroaryl, and heterocyclyl are as defined herein.

[0093] 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- wherein alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, heteroaryl, and heterocyclyl are as defined herein.

[0094] “Aminosulfonyl” refers to the group -SC>2NR21R22, wherein R21and R22independently are 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, substituted heterocyclic and where R21and R22are optionally joined together with the nitrogen bound thereto to form a heterocyclic or substituted heterocyclic group and alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic are as defined herein.

[0095] “Sulfonylamino” refers to the group -NR21SC>2R22, wherein R21and R22independently are 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 and where R21and R22are optionally joined together with the atoms bound thereto to form a heterocyclic or substituted heterocyclic group, and 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 are as defined herein.

[0096] “Aryl” or “Ar” refers to a monovalent aromatic carbocyclic group of from 6 to 18 carbon atoms having a single ring (such as is 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 condensed rings may or may not be aromatic, provided that the 23NAI-5011166969vlpoint of attachment is through an atom of an aromatic ring. This term includes, by way of example, phenyl and naphthyl. Unless otherwise constrained by the definition for the aryl substituent, such aryl groups can optionally be substituted with from 1 to 5 substituents, or from 1 to 3 substituents, selected from acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted alkoxy, substituted alkenyl, substituted alkynyl, substituted cycloalkyl, substituted cycloalkenyl, amino, substituted amino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, 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, -SCh-alkyl, -SCh-substituted alkyl, -SO2-aryl, -SCh-heteroaryl and trihalom ethyl.

[0097] “Aryloxy” refers to the group -O-aryl, wherein aryl is as defined herein, including, by way of example, phenoxy, naphthoxy, and the like, including optionally substituted aryl groups as also defined herein.

[0098] “Amino” refers to the group -NH2.

[0099] 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.

[0100] The term “azido” refers to the group -N3.

[0101] “Carboxyl,” “carboxy” or “carboxylate” refers to -CO2H or salts thereof.

[0102] “Carboxyl ester” or “carboxy ester” or the terms “carboxyalkyl” or “carboxylalkyl” refers to the groups -C(O)O-alkyl, -C(O)O-substitutedalkyl, -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-substitutedcycloalkenyl, -C(O)O-heteroaryl, -C(O)O-substituted heteroaryl, -C(O)O-heterocyclic, and -C(O)O-substituted heterocyclic, 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 are as defined herein.

[0103] “(Carboxyl ester)oxy” or “carbonate” refers to the groups -O-C(O)O-alkyl, -O-C(O)O-substituted alkyl, -O-C(O)O-alkenyl, -O-C(O)O-substituted alkenyl, -O- 24NAI-5011166969vlC(O)O-alkynyl, -O-C(O)O-substituted alkynyl, -O-C(O)O-aryl, -O-C(O)O-substituted aryl, -O-C(O)O-cycloalkyl, -O-C(O)O-substituted cycloalkyl, -O-C(O)O-cycloalkenyl, -O-C(O)O-substituted cycloalkenyl, -O-C(O)O-heteroaryl, -O-C(O)O-substituted heteroaryl, -O-C(O)O-heterocyclic, and -O-C(O)O-substituted heterocyclic, 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 are as defined herein.

[0104] “Cyano” or “nitrile” refers to the group -CN.

[0105] “Cycloalkyl” refers to cyclic alkyl groups of from 3 to 10 carbon atoms having single or multiple cyclic rings including fused, bridged, and spiro ring systems. Examples of suitable cycloalkyl groups include, for instance, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl and the like. Such cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like, or multiple ring structures such as adamantanyl, and the like.

[0106] The term “substituted cycloalkyl” refers to cycloalkyl groups having from 1 to 5 substituents, or from 1 to 3 substituents, selected from 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, -SCh-alkyl, -SCh-substituted alkyl, -SCh-aryl and -SCh-heteroaryl.

[0107] “Cycloalkenyl” refers to non-aromatic cyclic alkyl groups of from 3 to 10 carbon atoms having single or multiple rings and having at least one double bond and preferably from 1 to 2 double bonds.

[0108] The term “substituted cycloalkenyl” refers to cycloalkenyl groups having from 1 to 5 substituents, or from 1 to 3 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-25NAI-5011166969vlalkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SCh-alkyl, -SCh-substituted alkyl, -SCh-aryl and -SCh-heteroaryl.

[0109] “Cycloalkynyl” refers to non-aromatic cycloalkyl groups of from 5 to 10 carbon atoms having single or multiple rings and having at least one triple bond.

[0110] “Cycloalkoxy” refers to -O-cycloalkyl.

[0111] “Cycloalkenyloxy” refers to -O-cycloalkenyl.

[0112] “Halo” or “halogen” refers to fluoro, chloro, bromo, and iodo.

[0113] “Hydroxy” or “hydroxyl” refers to the group -OH.

[0114] “Heteroaryl” refers to an aromatic group of from 1 to 15 carbon atoms, such as from 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 (such as, pyridinyl, imidazolyl or furyl) or multiple condensed rings in a ring system (for example as in groups such as, indolizinyl, quinolinyl, benzofuran, benzimidazolyl or benzothienyl), wherein at least one ring within the ring system is aromatic. To satisfy valence requirements, any heteroatoms in such heteroaryl rings may or may not be bonded to H or a substituent group, e.g., an alkyl group or other substituent as described herein. In certain embodiments, the nitrogen and / or sulfur ring atom(s) of the heteroaryl group are optionally oxidized to provide for the N-oxide (N— >0), sulfinyl, or sulfonyl moieties. This term includes, by way of example, pyridinyl, pyrrolyl, indolyl, thiophenyl, and furanyl. Unless otherwise constrained by the definition for the heteroaryl substituent, such heteroaryl groups can be optionally substituted with 1 to 5 substituents, or from 1 to 3 substituents, selected from acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted alkoxy, substituted alkenyl, substituted alkynyl, substituted cycloalkyl, substituted cycloalkenyl, amino, substituted amino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, 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 trihalom ethyl.

[0115] The term “heteroaralkyl” refers to the groups -alkylene-heteroaryl where alkylene and heteroaryl are defined herein. This term includes, by way of example, pyridylmethyl, pyridylethyl, indolylmethyl, and the like.

[0116] “Heteroaryloxy” refers to -O-heteroaryl.26NAI-5011166969vl

[0117] “Heterocycle,” “heterocyclic,” “heterocycloalkyl,” and “heterocyclyl” refer to a saturated or unsaturated group having a single ring or multiple condensed rings, including fused bridged and spiro ring systems, and having from 3 to 20 ring atoms, including 1 to 10 hetero atoms. These ring atoms are selected from nitrogen, sulfur, or oxygen, where, in fused ring systems, one or more of the rings can be cycloalkyl, aryl, or heteroaryl, provided that the point of attachment is through the non-aromatic ring. In certain embodiments, the nitrogen and / or sulfur atom(s) of the heterocyclic group are optionally oxidized to provide for the N-oxide, -S(O)-, or -SO2- moieties. To satisfy valence requirements, any heteroatoms in such heterocyclic rings may or may not be bonded to one or more H or one or more substituent group(s), e.g., an alkyl group or other substituent as described herein.

[0118] Examples of heterocycles and heteroaryls include, but are not limited to, 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, isoxazole, 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 referred to as thiamorpholinyl), 1,1-dioxothiomorpholinyl, piperidinyl, pyrrolidine, tetrahydrofuranyl, and the like.

[0119] Unless otherwise constrained by the definition for the heterocyclic substituent, such heterocyclic groups can be optionally substituted with 1 to 5, or from 1 to 3 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, -SCh-alkyl, -SO2- substituted alkyl, -SCh-aryl, -SCh-heteroaryl, and fused heterocycle.

[0120] “Heterocyclyloxy” refers to the group -O-heterocyclyl.

[0121] The term “heterocyclylthio” refers to the group heterocyclic-S-.

[0122] The term “heterocyclene” refers to the diradical group formed from a heterocycle, as defined herein.27NAI-5011166969vl

[0123] The term “hydroxyamino” refers to the group -NHOH.

[0124] “Nitro” refers to the group -NO2.

[0125] “ Oxo” refers to the atom (=0).

[0126] “Sulfonyl” refers to the group -SO2-alkyl, -SO2-substituted alkyl, -SO2-alkenyl, -SO2- substituted alkenyl, -SO2-cycloalkyl, -SO2-substituted cycloalkyl, -SO2-cycloalkenyl, -SO2- substituted cycloalkenyl, -SO2-aryl, -SO2-substituted aryl, -SO2-heteroaryl, -SO2-substituted heteroaryl, -SO2-heterocyclic, and -SO2-substituted heterocyclic, 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 are as defined herein. Sulfonyl includes, by way of example, methyl-SO2-, phenyl-SO2-, and 4-methylphenyl-SO2-.

[0127] “Sulfonyloxy” refers to the group -OSCh-alkyl, -OSCh-substituted alkyl, -OSO2-alkenyl, -OSCh-substituted alkenyl, -OSCh-cycloalkyl, -OSCh-substituted cycloalkyl, -OSO2-cycloalkenyl, -OSCh-substituted cycloalkenyl, -OSO2-aryl, -OSO2-substituted aryl, -OSO2-heteroaryl, -OSO2-substituted heteroaryl, -OSO2-heterocyclic, and -OSO2-substituted heterocyclic, 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 are as defined herein.

[0128] “Sulfate” or “sulfate ester” refers the group -O-SO2-OH, -O-SO2-O-alkyl, -O-SO2-O-substituted alkyl, -O-SO2-O-alkenyl, -O-SO2-O-substituted alkenyl, -O-SO2-O-cycloalkyl, -O-SO2-O-substituted cycloalkyl, -O-SO2-O-cycloalkenyl, -O-SO2-O-substituted cycloalkenyl, -O-SO2-O-aryl, -O-SO2-O-substituted aryl, -O-SO2-O-heteroaryl, -O-SO2-O-substituted heteroaryl, -O-SO2-O-heterocyclic, and -O-SO2-O-substituted heterocyclic, 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 are as defined herein.

[0129] The term “aminocarbonyloxy” refers to the group -OC(O)NRR where each R is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, or heterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl and heterocyclic are as defined herein.

[0130] “ Thiol” refers to the group -SH.

[0131] “ Thioxo” or the term “thioketo” refers to the atom (=S).28NAI-5011166969vl

[0132] “Alkylthio” or the term “thioalkoxy” refers to the group -S-alkyl, wherein alkyl is as defined herein. In certain embodiments, sulfur may be oxidized to -S(O)-. The sulfoxide may exist as one or more stereoisomers.

[0133] The term “substituted thioalkoxy” refers to the group -S-substituted alkyl.

[0134] The term “thioaryloxy” refers to the group aryl-S- wherein the aryl group is as defined herein including optionally substituted aryl groups also defined herein.

[0135] The term “thioheteroaryl oxy” refers to the group heteroaryl-S- wherein the heteroaryl group is as defined herein including optionally substituted aryl groups as also defined herein.

[0136] The term “thioheterocyclooxy” refers to the group heterocyclyl-S- wherein the heterocyclyl group is as defined herein including optionally substituted heterocyclyl groups as also defined herein.

[0137] In the disclosure herein, the term “substituted,” when used to modify a specified group or radical, can also mean that one or more hydrogen atoms of the specified group or radical are each, independently of one another, replaced with the same or different substituent groups as defined below.

[0138] In addition to the groups disclosed with respect to the individual terms herein, substituent groups for substituting for one or more hydrogens (any two hydrogens on a single carbon can be replaced with =0, =NR70, =N-OR70, =N2 or =S) on saturated carbon atoms in the specified group or radical are, unless otherwise specified, -R60, halo,=0, -OR70, -SR70, -NR80R80, trihalomethyl, -CN, -OCN, -SCN, -NO, -NO2,=N2, -N3, -SO2R70, -SO2O M+, -SO2OR70, -OSO2R70, -OSO2O M+,-OSO2OR70, -P(O)(O )2(M+)2, -P(O)(OR70)O M+, -P(O)(OR70)2,-C(O)R70, -C(S)R70, -C(NR70)R70, -C(O)O M+, -C(O)OR70, -C(S)OR70, -C(O)NR80R80, -C(NR70)NR80R80, -OC(O)R70, -OC(S)R70, -0C(0)0M+, -OC(O)OR70, -OC(S)OR70, -NR70C (O)R70, -NR70C(S)R70, -NR70C02M , -NR70C02R70, -NR70C(S)OR70, -NR70C(O)NR80R80, -NR70C(NR70)R70and -NR70C(NR70)NR80R80, where R60is selected from the group consisting of optionally substituted alkyl, cycloalkyl, heteroalkyl, heterocycloalkylalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl and heteroarylalkyl, each R70is independently hydrogen or R60; each R80is independently R70or alternatively, two R80s, taken together with the nitrogen atom to which they are bonded, form a 5-, 6- or 7-membered heterocycloalkyl which may optionally include from 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 counter ion with a net single positive charge. Each M+may 29NAI-5011166969vlindependently be, for example, an alkali ion, such as K+, Na+, Li+; an ammonium ion, such as+N(R60)4; or an alkaline earth ion, such as [Ca2+]o.s, [Mg2+]o.s, or [Ba2+]o.s (“subscript 0.5 means that one of the counter ions for such divalent alkali earth ions can be an ionized form of a compound of the invention and the other a typical counter ion such as chloride, or two ionized compounds disclosed herein can serve as counter ions for such divalent alkali earth ions, or a doubly ionized compound of the invention can serve as the counter ion for such divalent alkali earth ions). As specific examples, -NR80R80is meant to include -NH2, -NH-alkyl, A-pyrrolidinyl, A-piperazinyl, 4 / f-methyl-piperazin-l-yl and 7V-morpholinyl.

[0139] In addition to the disclosure herein, substituent groups for hydrogens on unsaturated carbon atoms in “substituted” alkene, alkyne, aryl and heteroaryl groups are, unless otherwise specified, -R60, halo, -OM+, -OR70, -SR70, -S M , -NR80R80,trihalomethyl, -CF3, -CN, -OCN, -SCN, -NO, -NO2, -N3, -SO2R70, -SO3M+, -SO3R70, -OSO2R70, -OSO3 M+, -OSO3R70, -PO3-2(M+)2, -P(O)(OR70)O M+, -P(O)(OR70)2, -C(O)R70, -C(S)R70, -C(NR70)R70, -CO2 M+, -CO2R70, -C(S)OR70, -C(O)NR80R80, -C(NR70)NR80R80, -OC(O)R70, -OC(S)R70, -OCO2 M+, -OCO2R70, -OC(S)OR70, -NR70C(O)R70, -NR7OC(S)R70, -NR70C02 M , -NR70C02R70, -NR70C(S)OR70, -NR70C(O)NR80R80, -NR70C(NR70)R70and -NR70C(NR70)NR80R80, where R60, R70, R80and M+are as previously defined, provided that in case of substituted alkene or alkyne, the substituents are not -OM+, -OR70, -SR70, or -S M .

[0140] In addition to the groups disclosed with respect to the individual terms herein, substituent groups for hydrogens on nitrogen atoms in “substituted” heteroalkyl and cycloheteroalkyl groups are, unless otherwisespecified, -R60, -OM+, -OR70, -SR70, -S’M+, -NR80R80,trihalomethyl, -CF3, -CN, -NO, -NO2, -S(O)2R70, -S(O)2O M+, -S(O)2OR70, -OS(O)2R70, -OS(O)2O M+, -OS(O)2OR70, -P(O)(O )2(M+)2, -P(O)(OR70)O M+, -P(O)(OR70)(OR70), -C(O) R70, -C(S)R70, -C(NR70)R70, -C(O)OR70, -C(S)OR70, -C(O)NR80R80, -C(NR70)NR80R80, -OC(O)R70, -OC(S)R70, -OC(O)OR70, -OC(S)OR70, -NR70C(O)R70, -NR70C(S)R70, -NR70C(O)OR70, -NR70C(S)OR70, -NR70C(O)NR80R80, -NR70C(NR70)R70and -NR70C(NR70)NR80R80, where R60, R70, R80and M+are as previously defined.

[0141] In addition to the disclosure herein, in a certain embodiment, a group that is substituted has 1, 2, 3, or 4 substituents, 1, 2, or 3 substituents, 1 or 2 substituents, or 1 substituent.

[0142] It is understood that in all substituted groups defined above, polymers arrived at by defining substituents with further substituents to themselves (e.g., substituted aryl having a 30NAI-5011166969vlsubstituted aryl group as a substituent which is itself substituted with a substituted aryl group, which is further substituted by a substituted aryl group, etc.) are not intended for inclusion herein. In such cases, the maximum number of such substitutions is three. For example, serial substitutions of substituted aryl groups specifically contemplated herein are limited to substituted aryl-(substituted aryl)-substituted aryl.

[0143] Unless indicated otherwise, the nomenclature of substituents that are not explicitly defined herein are arrived at by naming the terminal portion of the functionality followed by the adjacent functionality toward the point of attachment. For example, the substituent “arylalkyloxycarbonyl” refers to the group (aryl)-(alkyl)-O-C(O)-.

[0144] As to any of the groups disclosed herein which contain one or more substituents, it is understood, of course, that such groups do not contain any substitution or substitution patterns which are sterically impractical and / or synthetically non-feasible. In addition, the subject compounds include all stereochemical isomers arising from the substitution of these compounds.

[0145] The term “pharmaceutically acceptable salt” means a salt which is acceptable for administration to a patient, such as a mammal (salts with counterions having acceptable mammalian safety for a given dosage regime). Such salts can be derived from pharmaceutically acceptable inorganic or organic bases and from pharmaceutically acceptable inorganic or organic acids. “Pharmaceutically acceptable salt” refers to pharmaceutically acceptable salts of a compound, which salts are derived from a variety of organic and inorganic counter ions well known in the art and include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like; and when the molecule contains a basic functionality, salts of organic or inorganic acids, such as hydrochloride, hydrobromide, formate, tartrate, besylate, mesylate, acetate, maleate, oxalate, and the like.

[0146] The term “salt thereof’ means a compound formed when a proton of an acid is replaced by a cation, such as a metal cation or an organic cation and the like. Where applicable, the salt is a pharmaceutically acceptable salt, although this is not required for salts of intermediate compounds that are not intended for administration to a patient. By way of example, salts of the present compounds include those wherein the compound is protonated by 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.

[0147] “ Solvate” refers to a complex formed by combination of solvent molecules with molecules or ions of the solute. The solvent can be an organic compound, an inorganic 31NAI-5011166969vlcompound, or a mixture of both. Some examples of solvents include, but are not limited to, methanol, -di methyl form am ide, tetrahydrofuran, dimethylsulfoxide, and water. When the solvent is water, the solvate formed is a hydrate.

[0148] “ Stereoisomer” and “stereoisomers” refer to compounds that have same atomic connectivity but different atomic arrangement in space. Stereoisomers include cis-trans isomers, E and Z isomers, enantiomers, and diastereomers.

[0149] “ Tautomer” refers to alternate forms of a molecule that differ only in electronic bonding of atoms and / or in the position of a proton, such as enol-keto and imine-enamine tautomers, or the tautomeric forms of heteroaryl groups containing a -N=C(H)-NH- ring atom arrangement, such as pyrazoles, imidazoles, benzimidazoles, triazoles, and tetrazoles. A person of ordinary skill in the art would recognize that other tautomeric ring atom arrangements are possible.

[0150] It will be appreciated that the term “or a salt or solvate or stereoisomer thereof’ is intended to include all permutations of salts, solvates and stereoisomers, such as a solvate of a pharmaceutically acceptable salt of a stereoisomer of subject compound.

[0151] “Pharmaceutically effective amount” and “therapeutically effective amount” refer to an amount of a compound sufficient to treat a specified disorder or disease or one or more of its symptoms and / or to prevent the occurrence of the disease or disorder. In reference to tumorigenic proliferative disorders, a pharmaceutically or therapeutically effective amount comprises an amount sufficient to, among other things, cause the tumor to shrink or decrease the growth rate of the tumor.

[0152] “Patient” refers to human and non-human subjects, especially mammalian subjects. The term “treating” or “treatment” as used herein means the treating or treatment of a disease or medical condition in a patient, such as a mammal (particularly a human) that includes: (a) preventing the disease or medical condition from occurring, such as, prophylactic treatment of a subject; (b) ameliorating the disease or medical condition, such as, eliminating or causing regression of the disease or medical condition in a patient; (c) suppressing the disease or medical condition, for example by, slowing or arresting the development of the disease or medical condition in a patient; or (d) alleviating a symptom of the disease or medical condition in a patient. In some embodiments, the term “treating,” or “treatment” excludes a prophylactic treatment.

[0153] The terms “polypeptide,” “peptide,” and “protein” are used interchangeably herein to refer to a polymeric form of amino acids of any length. Unless specifically indicated otherwise, “polypeptide,” “peptide,” and “protein” can include genetically coded and non- 32NAI-5011166969vlcoded amino acids, chemically or biochemically modified or derivatized amino acids, and polypeptides having modified peptide backbones. The term includes fusion proteins, including, but not limited to, fusion proteins with a heterologous amino acid sequence, fusions with heterologous and homologous leader sequences, proteins which contain at least one N-terminal methionine residue (e.g., to facilitate production in a recombinant host cell); immunologically tagged proteins; and the like. In certain embodiments, a polypeptide is an antibody.

[0154] “Native amino acid sequence” or “parent amino acid sequence” are used interchangeably herein to refer to the amino acid sequence of a polypeptide prior to modification to include at least one modified amino acid residue.

[0155] The terms “amino acid analog,” “unnatural amino acid,” and the like may be used interchangeably, and include amino acid-like compounds that are similar in structure and / or overall shape to one or more amino acids commonly found in naturally occurring proteins (e.g., Ala or A, Cys or C, Asp or D, Glu or E, Phe or F, Gly or G, His or H, He or I, Lys or K, Leu or L, Met or M, Asn or N, Pro or P, Gin or Q, Arg or R, Ser or S, Thr or T, Vai 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 with the same stereochemistry as in the naturally occurring D-form, as well as the L-form of amino acid analogs. In some instances, the amino acid analogs share backbone structures, and / or the side chain structures of one or more natural amino acids, with difference(s) being one or more modified groups in the molecule. Such modification may include, but is not limited to, substitution of an atom (such as N) for a related atom (such as S), addition of a group (such as methyl, or hydroxyl, etc.) or an atom (such as Cl or Br, etc.), deletion of a group, substitution of a covalent bond (single bond for double bond, etc.), or combinations thereof. For example, amino acid analogs may include a-hydroxy acids, and a-amino acids, and the like. Examples of amino acid analogs include, but are not limited to, sulfoalanine, and the like.

[0156] The terms “amino acid side chain” or “side chain of an amino acid” and the like may be used to refer to the substituent attached to the a-carbon of an amino acid residue, including natural amino acids, unnatural amino acids, and amino acid analogs. An amino acid side chain can also include an amino acid side chain as described in the context of the modified amino acids and / or conjugates described herein.

[0157] The term “carbohydrate” and the like may be used to refer to monomers units and / or polymers of monosaccharides, disaccharides, oligosaccharides, and polysaccharides. The term sugar may be used to refer to the smaller carbohydrates, such as monosaccharides,33NAI-5011166969vldi saccharides. The term “carbohydrate derivative” includes compounds where one or more functional groups of a carbohydrate of interest are substituted (replaced by any convenient substituent), modified (converted to another group using any convenient chemistry) or absent (e.g., eliminated or replaced by H). A variety of carbohydrates and carbohydrate derivatives are available and may be adapted for use in the subject compounds and conjugates.

[0158] The term “glycoside” or “glycosyl” refers to a sugar molecule or group bound to a moiety via a glycosidic bond. For example, the moiety that the glycoside is bound to can be a cleavable linker as described herein. A glycosidic bond can link the glycoside to the other moiety through various types of bonds, such as, but not limited to, an O-glycosidic bond (an O-glycoside), an N-glycosidic bond (a glycosylamine), an S-glycosidic bond (a thioglycoside), or C-glycosidic bond (a C-glycoside or C-glycosyl). In some cases, glycosides can be cleaved from the moiety they are attached to, such as by chemically-mediated hydrolysis or enzymatically-mediated hydrolysis.

[0159] 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, antibody fragments (e.g., Fab fragments), and the like. An antibody is capable of binding a target antigen. (Janeway, C., Travers, P., Walport, M., Shlomchik (2001) Immuno Biology, 5th Ed., Garland Publishing, New York). A target antigen can have one or more binding sites, also called epitopes, recognized by complementarity determining regions (CDRs) formed by one or more variable regions of an antibody.

[0160] The term “natural antibody” refers to an antibody in which the heavy and light chains of the antibody have been made and paired by the immune system of a multi-cellular organism. Spleen, lymph nodes, bone marrow and serum are examples of tissues that produce natural antibodies. For example, the antibodies produced by the antibody producing cells isolated from a first animal immunized with an antigen are natural antibodies.

[0161] The term “humanized antibody” or “humanized immunoglobulin” refers to a nonhuman (e.g., mouse or rabbit) antibody containing one or more amino acids (in a framework region, a constant region or a CDR, for example) that have been substituted with a correspondingly positioned amino acid from a human antibody. In general, humanized antibodies produce a reduced immune response in a human host, as compared to a nonhumanized version of the same antibody. Antibodies can be humanized using a variety of techniques known in the art including, for example, CDR-grafting (EP 239,400; PCT34NAI-5011166969vlpublication WO 91 / 09967; U.S. Pat. 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. Pat. No. 5,565,332). In certain embodiments, framework substitutions are identified by modeling of the interactions of the CDR and framework residues to identify framework residues important for antigen binding and sequence comparison to identify unusual framework residues at particular positions (see, e.g., U.S. Pat. No. 5,585,089; Riechmann et al., Nature 332:323 (1988)). Additional methods for humanizing antibodies contemplated for use in the present invention are described in U.S. Pat. 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, and PCT publications WO 98 / 45331 and WO 98 / 45332. In particular embodiments, a subject rabbit antibody may be humanized according to the methods set forth in US20040086979 and US20050033031. Accordingly, the antibodies described above may be humanized using methods that are well known in the art.

[0162] The term “chimeric antibodies” refer to antibodies whose light and heavy chain genes have been constructed, typically by genetic engineering, from antibody variable and constant region genes belonging to different species. For example, the variable segments of the genes from a mouse monoclonal antibody may be joined to human constant segments, such as gamma 1 and gamma 3. An example of a therapeutic chimeric antibody is a hybrid protein composed of the variable or antigen-binding domain from a mouse antibody and the constant or effector domain from a human antibody, although domains from other mammalian species may be used.

[0163] An immunoglobulin polypeptide immunoglobulin light or heavy chain variable region is composed of a framework region (FR) interrupted by three hypervariable regions, also called “complementarity determining regions” or “CDRs”. The extent of the framework region and CDRs have been defined (see, “Sequences of Proteins of Immunological Interest,” E. Kabat et al., U.S. Department of Health and Human Services, 1991). The framework region of an antibody, which is the combined framework regions of the constituent light and heavy chains, serves to position and align the CDRs. The CDRs are primarily responsible for binding to an epitope of an antigen.

[0164] A “parent Ig polypeptide” is a polypeptide comprising an amino acid sequence which lacks 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-35NAI-5011166969vlexisting amino acid sequence modifications (such as additions, deletions and / or substitutions).

[0165] As used herein the term “isolated” is meant to describe a compound of interest that is in an environment different from that in which the compound naturally occurs. “Isolated” is meant to include compounds that are within samples that are substantially enriched for the compound of interest and / or in which the compound of interest is partially or substantially purified.

[0166] As used herein, the term “substantially purified” refers to a compound that is 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 more than 98% free, from other components with which it is naturally associated.

[0167] The term “physiological conditions” is meant to encompass those conditions compatible with living cells, e.g., predominantly aqueous conditions of a temperature, pH, salinity, etc. that are compatible with living cells.

[0168] By “reactive partner” is meant a molecule or molecular moiety that specifically reacts with another reactive partner to produce a reaction product. Exemplary reactive partners include a cysteine or serine of a sulfatase motif and Formylglycine Generating Enzyme (FGE), which react to form a reaction product of a converted aldehyde tag containing a formylglycine (fGly) in lieu of cysteine or serine in the motif. Other exemplary reactive partners include an aldehyde of an fGly residue of a converted aldehyde tag (e.g., a reactive aldehyde group) and an “aldehyde-reactive reactive partner”, which comprises an aldehyde-reactive group and a moiety of interest, and which reacts to form a reaction product of a polypeptide having the moiety of interest conjugated to the polypeptide through the fGly residue.

[0169] “N-terminus” refers to the terminal amino acid residue of a polypeptide having a free amine group, which amine group in non-N-terminus amino acid residues normally forms part of the covalent backbone of the polypeptide.

[0170] “ C-terminus” refers to the terminal amino acid residue of a polypeptide having a free carboxyl group, which carboxyl group in non-C-terminus amino acid residues normally forms part of the covalent backbone of the polypeptide.

[0171] By “internal site” as used in referenced 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 at the C-terminus.36NAI-5011166969vl

[0172] The term “subject” refers to human and non-human subjects, especially mammalian subjects.

[0173] The terms “Tissue Factor,” “TF,” “platelet tissue factor,” “factor III,” “thromboplastin,” and “CD 142” are used interchangeably herein to refer to TF, or any variants (e.g., splice variants and allelic variants), isoforms, and species homologs of TF that are naturally expressed by cells, or that are expressed by cells transfected with a TF gene. In some aspects, the TF protein is a TF protein naturally expressed by a primate (e.g., a monkey or a human), a rodent (e.g., a mouse or a rat), a dog, a camel, a cat, a cow, a goat, a horse, a pig or a sheep. The term TF encompasses “full-length” TF, as well as any form of TF or any fragment thereof, for example those resulted from processing in a cell. In some embodiments, the TF comprises a signal sequence. In some embodiments, the TF does not include a signal sequence. In some embodiments, the term TF refers to a fragment of the full-length TF, which comprises a TF extracellular domain (ECD). The term TF also encompasses naturally occurring variants of TF, such as SNP variants, splice variants and allelic variants. In some aspects, the TF protein is human TF (hTF;METPAWPRVPRPETAVARTLLLGWVFAQVAGASGTTNTVAAYNLTWKSTNFKTILE WEPKPVNQVYTVQISTKSGDWKSKCFYTTDTECDLTDEIVKDVKQTYLARVFSYPA GNVESTGSAGEPLYENSPEFTPYLETNLGQPTIQSFEQVGTKVNVTVEDERTLVRRNN TFLSLRDVFGKDLIYTLYYWKSSSSGKKTAKTNTNEFLIDVDKGENYCFSVQAVIPSR TVNRKSTDSPVECMGQEKGEFREIFYIIGAVVFVVIILVIILAISLHKCRKAGVGQSWK ENSPLNVS (SEQ ID NO: 175) wherein the underline denotes signal peptide). In further embodiments, the human TF does not comprise a signal peptide, for example amino acid (aa) 33 to aa 295 of SEQ ID NO: 175. In yet further embodiments, the human TF as used herein refers to the extracellular domain (ECD) of the human TF, for example, aa 33 to aa 251 of SEQ ID NO: 175. In some aspects, the TF protein is cynomolgus TF (cTF;SGTTNTVAAYNLTWKSTNFKTILEWEPKPINQVYTVQISTKSGDWKSKCFYTADTEC DLTDEIVKDVKQTYLARVFSYPAGHVESTGSTEEPPYENSPEFTPYLETNLGQPTIQSF EQVGTKVNVTVQDEWTLVRRNDTFLSLRDVFGKDLIYTLYYWKSSSSGKKTAKTNT NEFLIDVDKGENYCFSVQAVIPSRRTANRKSTDSPVECMGHEKGESREIFYIIGAVVF VVIILVIILAISLHKCKKARVGRSWKENSPLNVA (SEQ ID NO: 176)). In further embodiments, the cynomolgus TF as used herein refers to its ECD, for example aa 1 to aa 220 of SEQ ID NO: 176. In some aspects, the TF protein is mouse TF (mTF;AGIPEKAFNLTWISTDFKTILEWQPKPTNYTYTVQISDRSRNWKNKCFSTTDTECDLT DEIVKDVTWAYEAKVLSVPRRNSVHGDGDQLVIHGEEPPFTNAPKFLPYRDTNLGQP37NAI-5011166969vlVIQQFEQDGRKLNVVVKDSLTLVRKNGTFLTLRQVFGKDLGYIITYRKGSSTGKKTN ITNTNEFSIDVEEGVSYCFFVQAMIFSRKTNQNSPGSSTVCTEQWKSFLGETLIIVGAV VLLATIFIILLSISLCKRRKNRAGQKGKNTPSRLA (SEQ ID NO: 177)). In further embodiments, the mouse TF as used herein refers to its ECD, for example aa 1 to aa 223 of SEQ ID NO: 177. In some aspects, the TF protein is pig TF (pTF;TGTTDVIVAYNLTWKSTNFKTILEWEPKPINYVYTVQISPRLGDWKNKCFHTTDTEC DVTDEIMRNVKETYVARVLSYPADTVLTAQEPPFTNSPPFTPYLDTNLGQPVIQSFEQ VGTKLNVTVEAARTLVRVNGTFLRLRDVFGKDLNYTLYYWRASSTGKKKATTNTN EFLIDVDKGENYCFSVQAVIPSRRVNQKSPESRIECTSQEKAVSRELFLIVGAVVFAVI VFVLVLSVSLYKCRKERAGPSGKENAPLNVA (SEQ ID NO: 178)). In some embodiments, the pig TF as used herein refers to its ECD, for example aa to aa 216 of SEQ ID NO: 178. TF is a cell surface receptor for the serine protease factor Vila. It is often times constitutively expressed by certain cells surrounding blood vessels and in some disease settings.

[0174] In some embodiments, the term TF as used herein refers to a TF epitope.

[0175] The terms “antibody,” “immunoglobulin,” or “Ig” are used interchangeably herein and are used in the broadest sense and specifically covers, for example polyclonal antibodies, monoclonal antibodies (including agonist, antagonist, neutralizing antibodies, 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 as well as antibody fragments (and / or polypeptides that comprise antibody fragments) that retain TF binding characteristics. Non-limiting examples of antibody fragments include antigenbinding regions and / or effector regions of the antibody, e.g., Fab, Fab’, F(ab’)2, Fv, scFv, (SCFV)2, single-chain antibody molecule, dual variable domain antibody, single variable domain antibody, linear antibody, V region, a multispecific antibody formed from antibody fragments, F(ab)2, Fd, Fc, diabody, di-diabody, disulfide-linked Fvs (dsFv), single-domain antibody (e.g., nanobody) or other fragments (e.g., fragments consisting of the variable regions of the heavy and light chains that are non-covalently coupled). In general terms, a variable (V) region can be any suitable arrangement of immunoglobulin heavy (VH) and / or light (VL) variable regions. For example, antibodies also include tetrameric antibodies comprising two heavy chain and two light chain molecules, an antibody light chain monomer,38NAI-5011166969vland an antibody heavy chain monomer. Thus, for example, the V region can be dimeric and contain VH-VH, VH-VL, or VL-VL dimers that bind TF. In any embodiment, a VH region and a VL region can be covalently coupled either directly or through a linker to form a single-chain Fv (scFv). For ease of reference, scFv proteins are referred to herein as included in the category “antibody fragments.” Another form of an antibody fragment is a peptide comprising one or more complementarity determining regions (CDRs) of an antibody. CDRs (also termed “minimal recognition units” or “hypervariable regions”) can be obtained by constructing polynucleotides that encode one or more CDRs of interest. Such polynucleotides are prepared, for example, by using the polymerase chain reaction to synthesize the variable region using mRNA of 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, for example, into single domain antibodies, maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, variable regions of new antigen receptors (v-NAR), 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 VL contain a light chain and / or a heavy chain constant region, such as one or more constant regions, including one or more IgGl, IgG2, IgG3 and / or IgG4 constant regions. In some embodiments, antibodies can include 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, IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2) of immunoglobulin molecule.

[0176] The term “monospecific,” as used herein denotes an antibody that has one or more binding sites each of which binds to the same epitope of the same antigen.

[0177] The term “bispecific” means that the antibody can specifically bind to at least two distinct antigenic determinants, for example two binding sites each formed by a pair of an antibody heavy chain variable region (VH) and an antibody light chain variable region (VL) binding to different antigens or to different epitopes on the same antigen. Such a bispecific antibody can have a 1+1 format (comprising one binding site for a first antigen or epitope and one binding site for a second antigen or epitope). Other bispecific antibody formats can be 2+1 or 1+2 formats (comprising two binding sites for a first antigen or epitope and one 39NAI-5011166969vlbinding site for a second antigen or epitope) or 2+2 format (comprising two binding sites for a first antigen or epitope and two binding sites for a second antigen or epitope). When a bispecific antibody comprises two antigen binding sites, each can bind to a different antigenic determinant. Such a bispecific antibody can bind to two different epitopes on the same antigen (e.g., epitopes on TF).

[0178] The terms “identical” or percent “identity” in the context of two or more nucleic acids or polypeptides, refer to two or more sequences or subsequences that are the same or have a specified percentage of nucleotides or amino acid residues that are the same, when compared and aligned (introducing gaps, if necessary) for maximum correspondence, not considering any conservative amino acid substitutions as part of the sequence identity. The 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 variants thereof. In some embodiments, two nucleic acids or polypeptides are substantially identical, 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, when compared and aligned for maximum correspondence, as measured using a sequence comparison algorithm or by visual inspection. In some embodiments, identity exists over a region of the amino acid sequences that is at least about ten residues, at least about 20 residues, at least about 40-60 residues, at least about 60-80 residues in length or any integral value there between. In some embodiments, identity exists over a longer region than 60-80 residues, such as at least about 80-100 residues, and in some embodiments, the sequences are substantially identical over the full-length of the sequences being compared, such as the coding region of a target protein or an antibody. In some embodiments, identity exists over a region of the nucleotide sequences that is at least about ten bases, at least about 20 bases, at least about 40-60 bases, at least about 60-80 bases in length or any integral value there between. In some embodiments, identity exists over a longer region than 60-80 bases, such as at least about 80-1000 bases or more, and in some embodiments, the sequences are substantially identical over the full length of the sequences being compared, such as a nucleotide sequence encoding a protein of interest.

[0179] A “conservative amino acid substitution” is one in which one amino acid residue is replaced with another amino acid residue having a side chain with similar chemical characteristics. Families of amino acid residues having similar side chains have been40NAI-5011166969vlgenerally defined in the art, including 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), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). For example, substitution of a phenylalanine for a tyrosine is a conservative substitution. Generally, conservative substitutions in the sequences of the polypeptides, soluble proteins, and / or antibodies of the disclosure do not abrogate the binding of the polypeptide, soluble protein, or antibody containing the amino acid sequence, to the target binding site. Methods of identifying amino acid conservative substitutions that do not eliminate binding are well known in the art.

[0180] As used herein, an “antigen” is a moiety or molecule that contains an epitope to which an antibody can bind. As such, an antigen can be bound by an antibody. In some embodiments, the antigen to which an antibody described herein binds is TF (e.g., human TF), or a fragment thereof, such as a fragment that comprises one or more regions of TF.

[0181] As used herein, an “epitope” is a term in the art and refers to a localized region of an antigen to which an antibody can bind. An epitope can be a linear epitope or a conformational, non-linear, or discontinuous epitope. In the case of a polypeptide antigen, for example, an epitope can be contiguous amino acids of the polypeptide (a “linear” epitope) or an epitope can comprise amino acids from two or more non-contiguous regions of the polypeptide (a “conformational,” “non-linear” or “discontinuous” epitope), e.g., human TF. It will be appreciated by one of skill in the art that, in general, a linear epitope may or may not be dependent 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 in a natural three-dimensional protein structure. In other embodiments, an antibody requires amino acid residues making up the epitope to exhibit a particular conformation (e.g., bend, twist, turn or fold) in order to recognize and bind the epitope.

[0182] An antibody binds “an epitope,” “essentially the same epitope,” or “the same epitope” as a reference antibody. The most widely used and rapid methods for determining whether two antibodies bind to identical, overlapping, or adjacent epitopes in a three-dimensional space are competition assays, which can be configured in a number of different formats, for example, 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 of41NAI-5011166969vlunlabeled antibodies to block the binding of labeled antibodies is measured using radioactive, fluorescent or enzyme labels.

[0183] “Epitope binning” is the process of grouping antibodies based on the epitopes they recognize. More particularly, epitope binning comprises methods and systems for discriminating the epitope recognition properties of different antibodies, using competition assays combined with computational processes for clustering antibodies based on their epitope recognition properties and identifying antibodies having distinct binding specificities. Additional details regarding methods for epitope binning and determining epitope binding of antibodies are described herein, as shown in Example 5.

[0184] As used herein, the terms “binds” “specifically binds,” “specifically recognizes,” “immunospecifically binds,” “selectively binds,” “immunospecifically recognizes” and “immunospecific” are analogous terms in the context of antibodies and refer to molecules that bind to an antigen (e.g., epitope) as understood by one skilled in the art.

[0185] In some embodiments, “specifically binds” means, for instance, that a polypeptide or molecule interacts more frequently, more rapidly, with greater duration, with greater affinity, or with some combination of the above to the epitope, protein, or target molecule than with alternative substances, including related and unrelated proteins. For example, a molecule that specifically binds to an antigen can bind to other peptides or polypeptides, generally with lower affinity as determined by, e.g, immunoassays, BIACORE™, KinExA 3000 instrument (Sapidyne Instruments, Boise, ID), the OctetQK384 system (ForteBio, Menlo Park, CA), or other assays known in the art. In some embodiments, an antibody or antigen binding region binds to or specifically binds to an antigen when it binds to the antigen with higher affinity than to any cross-reactive antigen as determined using experimental techniques, such as radioimmunoassays (RIAs) and enzyme linked immunosorbent assays (ELISAs). Typically, a specific or selective reaction will be at least twice background signal or noise and can be more than ten times background. See, e.g, Fundamental Immunology 332-36 (Paul ed., 2d ed. 1989) for a discussion regarding binding specificity. In some embodiments, the extent of binding of an antibody or antigen-binding region to a “nontarget” protein is less than about 10% of the binding of the antibody or antigen-binding region to its target antigen, for example, as determined by fluorescence activated cell sorting (FACS) analysis or RIAs. In some embodiments, molecules that specifically bind to an antigen bind to the antigen with a KA that is at least 2 logs, 2.5 logs, 3 logs, 4 logs or greater than the KA when the molecules bind to another antigen. In some embodiments, molecules that specifically bind to an antigen do not cross react with other proteins. In another specific 42NAI-5011166969vlembodiment, molecules that specifically bind to an antigen do not cross react with other non-TF proteins. In some embodiments, “specifically binds” means, for instance, that a polypeptide or molecule binds a protein or target with a KD of about 0.1 mM or less, but more usually less than about 1 pM. In some embodiments, “specifically binds” means that a polypeptide or molecule binds a target with a KD of at least about 0.1 pM or less, at least about 0.01 pM or less, or at least about 1 nM or less. Because of the sequence identity between homologous proteins in different species, specific binding can include a polypeptide or molecule that recognizes a protein or target in more than one species. Likewise, because of homology within certain regions of polypeptide sequences of different proteins, specific binding can include a polypeptide or molecule that recognizes more than one protein or target. It is understood that, in some embodiments, a polypeptide or molecule that specifically binds a first target may or may not specifically bind a second target. As such, “specific binding” does not necessarily require (although it can include) exclusive binding, e.g. , binding to a single target. Thus, in some embodiments, a polypeptide or molecule can specifically bind more than one target. In some embodiments, multiple targets can be bound by the same antigen-binding site on the polypeptide or molecule. For example, in some embodiments, an antibody can comprise two identical antigen-binding sites, each of which specifically binds the same epitope on two or more proteins. In alternative embodiments, an antibody can be bispecific and comprise at least two antigen-binding sites with differing specificities. Generally, but not necessarily, reference to “binding” means “specific binding.”

[0186] The term “binding affinity” generally refers to the strength of the sum of noncovalent interactions between a single binding site of a molecule (e.g., antibody) and its binding partner (e.g., an antigen such as TF). Unless indicated otherwise, as used herein, “binding affinity” refers to intrinsic binding affinity that reflects a 1 : 1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of a binding molecule X for its binding partner Y can generally be represented by the dissociation constant (KD). Affinity can be measured by common methods known in the art, including those described herein. Low-affinity antibodies generally bind antigen slowly and tend to dissociate readily, whereas high-affinity antibodies generally bind antigen faster and tend to remain bound longer. A variety of methods of measuring binding affinity is known in the art, any of which can be used for purposes of the present disclosure. In one embodiment, the “KD” or “KD value” can be measured by biolayer interferometry (BLI) using, for example, the OctetQK384 system (ForteBio, Menlo Park, CA). Alternatively, the KD may also be measured in a radiolabeled antigen binding assay (RIA), for example, performed with the Fab 43NAI-5011166969vlversion of an antibody of interest and its antigen (Chen et al., (1999) J. Mol Biol 293:865-881) or using surface plasmon resonance (SPR) assays by BIACORE™, using, for example, a BIACORE™-2000 or a BIACORE™-3000 (BIAcore, Inc., Piscataway, NJ). An “on-rate” or “rate of association” or “association rate” or “kon,” as well as an “off-rate” or “rate of dissociation” or “dissociation rate” or “koff,” can also be determined with the same SPR or BLI techniques described herein using, for example, the OctetQK384 system (ForteBio, Menlo Park, CA) or a BIACORE™-2000 or a BIACORE™-3000 (BIACORE™, Inc., Piscataway, NJ), respectively.

[0187] The term “compete,” when used in the context of a TF antibody, describes a binding agent that, in the presence of another binding agent, is at least partially inhibited from binding to an epitope or binding site due to binding of the other binding agent. Competition can be determined by an assay in which the binding agent under study prevents or inhibits the specific binding of a reference molecule (e.g., a reference ligand, or reference antigen binding protein, such as a reference antibody) to a common antigen (e.g, TF). Numerous types of competitive binding assays can be used to determine if a test binding agent competes with a reference molecule for binding to TF (e.g, human TF). Examples of assays that can be employed include solid phase direct or indirect radioimmunoassay (RIA), solid phase direct or indirect enzyme immunoassay (EIA), sandwich competition assay (see, e.g., Stahli 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 labeled assay, solid phase direct labeled sandwich assay (see, e.g, Harlow and Lane, (1988) Antibodies, A Laboratory Manual, Cold Spring Harbor Press); solid phase direct label RIA using 1-125 label (see, e.g, Morel et al., (1988) Molec. Immunol. 25:7-15); and direct labeled RIA (Moldenhauer et al., (1990) Scand. J. Immunol.32:77-82). Typically, such an assay involves the use of a purified antigen (e.g., TF, such as human TF) bound to a solid surface or cells bearing either of an unlabeled test antigen binding protein (e.g., test TF antibody or ADC) or a labeled reference antigen binding protein (e.g., reference TF antibody or ADC). Competitive inhibition can be measured by determining the amount of label bound to the solid surface or cells in the presence of the test antigen binding protein. Usually, the test antigen binding protein is present in excess.Antibodies identified by competition assay (competing antibodies) include antibodies binding to the same epitope as the reference antibody and / or antibodies binding to an adjacent epitope sufficiently proximal to the epitope bound by the reference for antibodies steric hindrance to occur (e.g., similar epitope or overlapping epitope). Usually, when a competing antibody is 44NAI-5011166969vlpresent in excess, it will inhibit specific binding of a reference antibody to a common antigen by at least 20%, for example, 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.

[0188] The terms “constant region” and “constant domain” are used interchangeably herein, are well-known antibody terms of art, and refer to an antibody portion, for example, a carboxyl terminal portion of a light and / or heavy chain which is not directly involved in binding of an antibody to an antigen, but which can exhibit various effector functions, such as interaction with an Fc receptor. The term includes the portion of an immunoglobulin molecule having a generally more conserved amino acid sequence relative to an immunoglobulin variable region.

[0189] Antibody “effector functions” refer to those biological activities attributable to the Fc region (e.g., a native sequence Fc region or amino acid sequence variant Fc region) of an antibody and which vary with the antibody isotype. Examples of antibody effector functions include: Clq binding and complement dependent cytotoxicity; Fc receptor binding; antibodydependent cell-mediated cytotoxicity (ADCC); phagocytosis; down regulation of cell surface receptors (e.g., B cell receptor); and B cell activation.

[0190] The term “Fc region” herein is used to define a 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 might vary, the human IgG heavy chain Fc region is often defined to stretch from an amino acid residue at position Cys226 (of the EU numbering system) or from Pro230 (of the EU numbering system) to the carboxyl-terminus thereof. The C-terminal lysine (residue 447 of the EU numbering system) of the Fc region can be removed, for example, during production or purification of the antibody, or by recombinantly engineering the nucleic acid encoding a heavy chain of the antibody. An exemplary Fc region sequence is provided below (CH2 domain = bold text; CH3 domain = underline text):CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVD GVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGOPREPQVYTLPPSRDELTKNOVSLTCLVKGFYPSDIAVEWESNGOPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWOOGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 76).

[0191] A “functional Fc region” possesses an “effector function” of a native sequence Fc region. Exemplary “effector functions” include Clq binding; complement dependent45NAI-5011166969vlcytotoxicity (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 generally require the Fc region to be combined with a binding region or binding domain e.g., an antibody variable region or domain) and can be assessed using various assays as disclosed.

[0192] A “native sequence Fc region” comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature, and not manipulated, modified, and / or changed (e.g., isolated, purified, selected, including, or combining with other sequences such as variable region sequences) by a human. Native sequence human Fc regions include a native sequence human IgGl Fc region (non- A and A allotypes); native sequence human IgG2 Fc region; native sequence human IgG3 Fc region; and native sequence human IgG4 Fc region; as well as naturally occurring variants thereof.

[0193] A “variant Fc region” comprises an amino acid sequence that differs from that of a native sequence Fc region by virtue of at least one amino acid modification, (e.g., substituting, addition, or deletion) preferably one or more amino acid substitution(s). In some embodiments, the variant Fc region has at least one amino acid substitution compared to a native sequence Fc region or to the Fc region of a parent polypeptide, for example, from about one to about ten amino acid substitutions, and preferably from about one to about five amino acid substitutions in a native sequence Fc region or in the Fc region of the parent polypeptide. The variant Fc region described herein can possess at least about 80% homology with a native sequence Fc region and / or with an Fc region of a parent polypeptide, or at least about 90% homology therewith, for example, at least about 95% homology therewith. The variant Fc region herein described herein can have a loss of effector function (e.g., silent Fc).

[0194] The term “heavy chain” when used in reference to an antibody refers to a polypeptide chain of about 50-70 kDa, wherein the amino-terminal portion includes a variable region of about 120 to 130 or more amino acids, and a carboxy -terminal portion includes one or more constant regions. The “heavy chain” can refer to any distinct types, e.g., for example, alpha (a), delta (8), epsilon (a), gamma (y) and mu (p), based on the amino acid sequence of the constant region, which give rise to IgA, IgD, IgE, IgG and IgM classes of antibodies, respectively, including subclasses of IgG, e.g., IgGl, IgG2, IgG3 and IgG4.

[0195] As used herein, the term “light chain” when used in reference to an antibody can refer to a polypeptide chain of about 25 kDa, wherein the amino-terminal portion includes a variable region of about 100 to about 110 or more amino acids, and a carboxy -terminal portion includes a constant region. The approximate length of a light chain is 211 to 21746NAI-5011166969vlamino acids. There are two distinct types, e.g., kappa (K) or lambda (1) based on the amino acid sequence of the constant regions. Light chain amino acid sequences are well known in the art.

[0196] The terms “antigen binding fragment,” “antigen binding domain,” “antigen binding region,” and similar terms refer to a portion of an antibody that comprises amino acid residues that interact with an antigen and confer on the binding fragment or region its specificity and affinity for the antigen (e.g., the CDRs). “Antigen binding fragment” as used herein includes “antibody fragment,” which comprises a portion of an antibody including one or more CDRs, such as the antigen binding or variable region of the antibody.

[0197] Antibodies described herein include, but are not limited to, synthetic antibodies, monoclonal antibodies, recombinantly produced antibodies, multispecific antibodies (e.g., including bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, intrabodies, single-chain Fvs (scFv) (e.g., including monospecific, bispecific, and the like), camelized antibodies, Fab fragments, F(ab’) fragments, disulfide-linked Fvs (sdFv), anti-idiotypic (anti-Id) antibodies, and epitope-binding fragments of any of the above.

[0198] In some embodiments, antibodies described herein include immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, such as molecules that contain one or more antigen binding sites that bind to TF.

[0199] A TF antibody, as described herein, can be of any type (e.g., IgG, IgE, IgM, IgD, IgA or IgY), any class (e.g., IgGl, IgG2, IgG3, IgG4, IgAl or IgA2), or any subclass (e.g., IgG2a or IgG2b) of immunoglobulin molecule. In some embodiments, a TF antibody, as described herein, is an IgG antibody (e.g., human IgG), or a class (e.g., human IgGl, IgG2, IgG3, or IgG4) or a subclass thereof.

[0200] In some embodiments, a TF antibody is a 4-chain antibody unit comprising two heavy (H) chain / 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 and / or L chains comprise constant regions, for example, human constant regions. In some embodiments, the L chain constant region of a TF antibody is a kappa or lambda light chain constant region, for example, a human kappa or lambda light chain constant region. In some embodiments, the H chain constant region of a TF antibody comprises a gamma heavy chain constant region, for example, a human gamma heavy chain constant region. In some embodiments, a TF antibody comprise an IgG constant region, for example, human IgG constant regions (e.g., IgGl, IgG2, IgG3, and / or IgG4 constant regions).47NAI-5011166969vl

[0201] 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 can preferentially bind to TF, such as human TF, which means that the antibody or fragment thereof binds to TF, such as human TF, with greater affinity than it binds to an unrelated control protein. For example, the antibody or fragment thereof can specifically recognize and bind to TF or a portion thereof. “Specific binding” means that the 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 greater than the affinity for an unrelated control protein (e.g, hen egg white lysozyme). In some embodiments, the TF antibody or fragment thereof can bind TF substantially exclusively (e.g., is able to distinguish TF from other known polypeptides, for example, by virtue of measurable differences in binding affinity). In some embodiments, a TF antibody can react with TF sequences other than human TF sequences (e.g., cynomolgus TF sequences).

[0202] The terms “variable region” and “variable domain” are used interchangeably to refer to a portion of the light and heavy chains of an antibody that are generally located at the amino-terminal of the light and heavy chain, has a length of about 120 to 130 amino acids in the heavy chain, about 100 to 110 amino acids in the light chain, and is used in the binding and specificity of each antibody for 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 certain segments of the variable regions differ extensively in sequence among antibodies. The V region mediates antigen binding and defines specificity of a particular antibody for its antigen. However, the variability is not evenly distributed across the 110-amino acid span of the variable regions. Instead, the V regions consist of less variable (e.g., relatively invariant) stretches called framework regions (FRs) of about 15-30 amino acids separated by shorter regions of greater variability (e.g., extreme variability) called “hypervariable regions” or alternatively called “complementarity determining regions.” The variable regions of heavy and light chains each comprise four frameworks (FR1, FR2, FR3 and FR4), largely adopting a P sheet configuration, connected by three hypervariable regions, which form loops connecting, and in some cases forming part of, the P sheet structure. The hypervariable regions in each chain are held together in proximity by the frameworks and, with the hypervariable regions from the other chain, contribute to 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 involved directly in 48NAI-5011166969vlbinding an antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody dependent cellular cytotoxicity (ADCC) and complement dependent cytotoxicity (CDC). The variable regions differ extensively in sequence between different antibodies. The variability in sequence is concentrated in the CDRs while the less variable portions in the variable region are referred to as framework regions (FR). The CDRs of the light and heavy chains are primarily responsible for the interaction of the antibody with antigen. In specific embodiments, the variable region is a human variable region.

[0203] The term “hypervariable region,” “HVR,” “HV,” “complementarity determining region, “and “CDR” when used herein refers to the regions of an antibody variable region that are hypervariable in sequence and / or form structurally defined loops. Generally, antibodies comprise six hypervariable regions: three in the VH (Hl or VH CDR1, H2 or VH CDR2, and H3 or VH CDR3), and three in the VL (LI or VL CDR1, L2 or VL CDR2, and L3 or VL CDR3). Several hypervariable region delineations are in use and are encompassed herein. The Kabat CDRs are based on sequence variability and are the most 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 refers instead to the location of the structural loops (see, e.g., Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). The end of the Chothia CDR-H1 loop when numbered using the Kabat numbering convention varies between H32 and H34 depending on the length of the loop (this is because the Kabat numbering scheme places the insertions at H35A and H35B; if neither 35 A nor 35B is present, the loop ends at 32; if only 35A is present, the loop ends at 33; if both 35A and 35B are present, the loop ends at 34). The AbM hypervariable regions represent a compromise between the Kabat CDRs and Chothia structural loops, and are used by Oxford Molecular’s AbM antibody modeling software (see, e.g., Martin, in Antibody Engineering, Vol. 2, Chapter 3, Springer Verlag). The “contact” hypervariable regions are based on an analysis of the available complex crystal structures. The residues from each of these hypervariable regions or CDRs are noted below.

[0204] A universal numbering system has been developed and widely adopted, ImMunoGeneTics (IMGT®) Information System (Lefranc et al., Dev. Comp. Immunol. 27(l):55-77 (2003)). IMGT® is an integrated information system specializing in immunoglobulins (IG), T cell receptors (TR) and major histocompatibility complex (MHC) of human and other vertebrates. Herein, the CDRs are referred to in terms of both the amino acid sequence and the location within the light or heavy chain. As the “location” of the CDRs within the structure of the immunoglobulin variable region is conserved between species and 49NAI-5011166969vlpresent in structures called loops, by using numbering systems that align variable region sequences of structural features, CDR and framework residues and are readily identified. This information can be used in grafting and replacement of CDR residues from immunoglobulins of one species into an acceptor framework from, typically, a human antibody. An additional numbering system (AHon) has been developed by Honegger and Pliickthun, J. Mol. Biol. 309: 657-670 (2001). Correspondence between the numbering system, including, for example, the Kabat numbering and the IMGT® unique numbering system, is well known to one skilled in the art (see, e.g., Kabat, supra, Chothia and Lesk, supra, Martin, supra, Lefranc et al., supra) and is also illustrated below. Various systems known in the art or described herein represent different ways of delineating CDRs, and when they are used to define the same antibody, they are often considered equivalent. An exemplary system, shown herein, combines Kabat and Chothia.Exemplary IMGT® Kabat AbM Chothia Contact VHCDR1 26-35 27-38 31-35 26-35 26-32 30-35 VHCDR2 50-65 56-65 50-65 50-58 53-55 47-58 VHCDR3 95-102 105-117 95-102 95-102 96-101 93-101 VLCDR1 24-34 27-38 24-34 24-34 26-32 30-36 VLCDR2 50-56 56-65 50-56 50-56 50-52 46-55VLCDR3 89-97 105-117 89-97 89-97 91-96 89-96

[0205] Hypervariable regions can comprise “extended hypervariable regions” as follows: 24-36 or 24-34 (LI), 46-56 or 50-56 (L2) and 89-97 or 89-96 (L3) in the VL and 26-35 or 26-35A (Hl), 50-65 or 49-65 (H2) and 93-102, 94-102, or 95-102 (H3) in the VH. As used herein, the terms “hypervariable region,” “HVR,” “HV,” “complementarity determining region,” or “CDR” are used interchangeably.

[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, such as TF-expressing tumor cells. A TF-mediated disease includes a cancer including, but not limited to, cancers that express or overexpress TF.

[0207] The term “tumor,” in any embodiment herein, refers to any neoplastic cell growth or proliferation, whether malignant or benign, and to 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.50NAI-5011166969vl

[0209] The term “ADC” refers to an antibody-drug conjugate, which in the context of the present invention refers to a TF antibody, which is coupled to another moiety which includes a drug, as described herein.

[0210] As used herein, “drug” refers to a compound that has biological activity, such as a cytotoxic compound (e.g., a cytotoxic small molecule, a cytotoxic synthetic peptide, and the like).

[0211] Examples of drugs include small molecule drugs, such as a cancer chemotherapeutic agent. For example, where the polypeptide is an antibody (or fragment thereof) that has specificity for a tumor cell, the antibody can be modified as described herein to include a modified amino acid, which can be subsequently conjugated to a cancer chemotherapeutic agent. Cancer chemotherapeutic agents include non-peptidic ( / .< ., non-proteinaceous) compounds that reduce proliferation of cancer cells, and encompass cytotoxic agents and cytostatic agents. Non-limiting examples of chemotherapeutic agents include alkylating agents, nitrosoureas, antimetabolites, antitumor antibiotics, plant (vinca) alkaloids, and steroid hormones. Peptidic compounds can also be used.

[0212] Suitable cancer chemotherapeutic agents include dolastatin and active analogs and derivatives thereof; and auristatin and active analogs and derivatives thereof (e.g., Monomethyl auristatin D (MM D), monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), and the like). See, e.g, WO 96 / 33212, WO 96 / 14856, and U.S.6,323,315. For example, dolastatin 10 or auristatin PE can be included in a TF-ADC of the present disclosure. Suitable cancer chemotherapeutic agents also include maytansinoids and active analogs and derivatives thereof (see, e.g, EP 1391213; and Liu et al (1996) Proc. Natl. Acad. Set. USA 93:8618-8623); duocarmycins and active analogs and derivatives thereof (e.g., including the synthetic analogues, KW-2189 and CB 1-TM1); and benzodiazepines and active analogs and derivatives thereof (e.g., pyrrol obenzodiazepine (PBD)).

[0213] Agents that act to reduce cellular proliferation are known in the art and widely used. Such agents include alkylating agents, such as nitrogen mustards, nitrosoureas, ethylenimine derivatives, alkyl sulfonates, 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] Antimetabolite agents include folic acid analogs, pyrimidine analogs, purine analogs, and adenosine deaminase inhibitors, including, but not limited to, cytarabine51NAI-5011166969vl(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 their derivatives, e.g., vinca alkaloids, antitumor antibiotics, enzymes, lymphokines, and epipodophyllotoxins), include, but are not limited to, Ara-C, paclitaxel (TAXOL®), docetaxel (TAXOTERE®), deoxy coformycin, mitomycin-C, L-asparaginase, azathioprine; brequinar; alkaloids, e.g. vincristine, vinblastine, vinorelbine, vindesine, and the like; podophyllotoxins, e.g. etoposide, teniposide, and the like; antibiotics, e.g. anthracycline, daunorubicin hydrochloride (daunomycin, rubidomycin, cerubidine), idarubicin, doxorubicin, epirubicin and morpholino derivatives, and the like; phenoxizone biscyclopeptides, e.g. dactinomycin; basic glycopeptides, e.g. bleomycin; anthraquinone glycosides, e.g. plicamycin (mithramycin); anthracenediones, e.g. mitoxantrone; azirinopyrrolo indolediones, e.g. mitomycin; macrocyclic immunosuppressants, e.g. cyclosporine, FK-506 (tacrolimus, prograf), rapamycin, and the like; and the like.

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

[0217] Microtubule affecting agents that have antiproliferative activity are also suitable for use and include, but are 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 cysterin, vinblastine sulfate, vincristine sulfate, natural and synthetic epothilones including but not limited to, eopthilone A, epothilone B, discodermolide; estramustine, nocodazole, and the like.

[0218] Hormone modulators and steroids (including synthetic analogs) that are suitable for use include, but are not limited to, adrenocorticosteroids, e.g. prednisone, dexamethasone, and the like; estrogens and pregestins, e.g. hydroxyprogesterone caproate, medroxyprogesterone acetate, megestrol acetate, estradiol, clomiphene, tamoxifen; and the like; adrenocortical suppressants, e.g. aminoglutethimide; 17a-ethinylestradiol; diethylstilbestrol, testosterone, fluoxymesterone, dromostanolone propionate, testolactone, methylprednisolone, methyl-testosterone, prednisolone, triamcinolone, chlorotrianisene, hydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesterone acetate, leuprolide, flutamide (DROGENIL®), toremifene (FARESTON®), and goserelin52NAI-5011166969vl(ZOLADEX®), and the like. Estrogens stimulate proliferation and differentiation; therefore, compounds that bind to the estrogen receptor are used to block this activity. Corticosteroids can inhibit T cell proliferation.

[0219] Other suitable chemotherapeutic agents include metal complexes, e.g. cisplatin (cis-DDP), carboplatin, and the like; ureas, e.g. hydroxyurea; hydrazines, e.g. N-m ethylhydrazine; epidophyllotoxin; a topoisomerase inhibitor; procarbazine; mitoxantrone; leucovorin; tegafur; and the like Other anti-proliferative agents of interest include immunosuppressants, e.g. mycophenolic acid, thalidomide, desoxyspergualin, azasporine, leflunomide, mizoribine, azaspirane (SKF 105685); gefitinib (IRESSA®, ZD 1839, 4-(3-chloro-4-fluorophenylamino)-7-methoxy-6-(3-(4-morpholinyl)propoxy)quinazoline); and the like.

[0220] Taxanes are suitable for use. “Taxanes” include paclitaxel, as well as any active taxane derivative or pro-drug. “Paclitaxel” (which should be understood herein to include analogues, formulations, and derivatives such as, for example, docetaxel, TAXOL®, TAXOTERE® (a formulation of docetaxel), 10-desacetyl analogs of paclitaxel and 3’N-desbenzoyl-3’N-t-butoxycarbonyl analogs of paclitaxel) can be readily prepared utilizing techniques known to those skilled in the art (see also WO 94 / 07882, WO 94 / 07881, WO 94 / 07880, WO 94 / 07876, WO 93 / 23555, WO 93 / 10076; U.S. Pat. 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 obtained from a variety of commercial sources, including for example, Sigma Chemical Co., St. Louis, Mo. (T7402 from Taxus brevifoHa or T-1912 from Taxus yannanensis). Paclitaxel should be understood to refer to not only the common chemically available form of paclitaxel, but analogs and derivatives (e.g., TAXOTERE® docetaxel, as noted herein) and paclitaxel conjugates (e.g., paclitaxel -PEG, paclitaxel-dextran, or paclitaxel -xylose).

[0221] Also included within the term “taxane” are a variety of known derivatives, including both hydrophilic derivatives, and hydrophobic derivatives. Taxane derivatives include, but are not limited to, galactose and mannose derivatives described in International Patent Application No. WO 99 / 18113; piperazino and other derivatives described in WO 99 / 14209; taxane derivatives described in WO 99 / 09021, WO 98 / 22451, and U.S. Patent No. 5,869,680; 6-thio derivatives described in WO 98 / 28288; sulfenamide derivatives described in U.S. Patent No. 5,821,263; and taxol derivative described in U.S. Patent No. 5,415,869. It further includes prodrugs of paclitaxel including, but not limited to, those described in WO 98 / 58927; WO 98 / 13059; and U.S. Patent 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;53NAI-5011166969vl(3) tumor-associated antigen antagonists, such as antibodies that bind specifically to a tumor antigen; (4) apoptosis receptor agonists; (5) interleukin-2; (6) IFN-a; (7) IFN-y; (8) colonystimulating factors; and (9) inhibitors of angiogenesis.

[0223] An “effective amount” is generally an amount sufficient to reduce the severity and / or frequency of symptoms, eliminate the symptoms and / or underlying cause, prevent the occurrence of symptoms and / or their underlying cause, and / or improve or remediate the damage that results from or is associated with a disease, disorder, or condition. In some embodiments, the effective amount is a therapeutically effective amount.

[0224] The term “therapeutically effective amount” as used herein refers to the amount of an antibody or ADC described herein that is sufficient to reduce and / or ameliorate the severity and / or duration of a given disease, disorder, or condition, and / or a symptom related thereto. A therapeutically effective amount of an agent, including a therapeutic agent, can be an amount necessary for (i) reduction or amelioration of the advancement or progression of a given disease, disorder, or condition, (ii) reduction or amelioration of the recurrence, development or onset of a given disease, disorder or conditions, and / or (iii) to improve or enhance the therapeutic effect of another therapy (e.g., a therapy other than the administration of an antibody or ADC described herein). A “therapeutically effective amount” of a substance / molecule / agent of the present disclosure (e.g., a TF antibody or ADC) can vary based on a number of factors such as the disease state, age, sex, and weight of the individual, and the ability of the substance / molecule / agent, to elicit a desired response in the individual. A therapeutically effective amount encompasses an amount in which any toxic or detrimental 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 affecting agent that has antiproliferative activity, such as a maytansinoid. In some embodiments, the drug is an antimitotic agent, such as an auristatin or an active auristatin analog or derivative thereof. In some embodiments, the drug is a DNA alkylating agent.

[0226] The term “pharmaceutically acceptable” as used herein means being approved by a regulatory agency of the federal or a state government, or listed in the U.S. Pharmacopeia, European Pharmacopeia, or other generally recognized Pharmacopeia for use in animals, and more particularly in humans.54NAI-5011166969vl

[0227] “Excipients” include carriers, excipients, preservatives, or stabilizers that are nontoxic to the cell or mammal being exposed thereto at the dosages and concentrations employed and can be included, for example, to affect stability, bulk up formulations, or to confer a therapeutic enhancement on the active ingredient in the final dosage form (e.g., facilitating absorption, reducing viscosity, enhancing solubility). An "excipient" can be an organic or inorganic ingredient, natural or synthetic with which the active ingredient is combined to facilitate the use of the active ingredient, e.g., the administration of the active ingredient to a subject. Examples of excipients include buffers such as phosphate, citrate, and other organic acids; antioxidants such as ascorbic acid; low molecular weight (e.g., less than about ten amino acid residues) polypeptide; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates such as glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as 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 with which the therapeutic is administered. Such excipients can be sterile liquids, such as 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 a composition (e.g., a pharmaceutical composition) is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed 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, dried skim milk, glycerol, propylene, glycol, water, ethanol, and the like. The composition, in any embodiment, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. Compositions can take the form of solutions, suspensions, emulsion, tablets, pills, capsules, powders, sustained-release formulations, and the like. Oral compositions, such as formulations, can include standard excipients such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, and the like. Examples of suitable excipients are described in Remington: The Science and Practice of Pharmacy (2020) (Elsevier Science, Amsterdam, Netherlands), including pharmaceutical compounds, can contain an effective amount or therapeutically effective amount of a TF-ADC, for example,55NAI-5011166969vlin isolated or purified form, together with a suitable amount of excipient to provide the form for proper administration to the subject. The formulation should suit 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 to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0229] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that 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 are also encompassed within the invention, subject to any specifically excluded limit in the stated range. 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 is 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 sub-combination. All combinations of the embodiments pertaining to the invention are specifically embraced by the present invention and are disclosed herein just as if each and every combination was individually and explicitly disclosed, to the extent that such combinations embrace subject matter that are, for example, compounds that are stable compounds ( / .< ., compounds that can be made, isolated, characterized, and tested for biological activity). In addition, all sub-combinations of the various embodiments and elements thereof (e.g., elements of the chemical groups listed in the embodiments describing such variables) are also specifically embraced by the present invention and are disclosed herein just as if each and every such sub-combination was individually and explicitly disclosed herein.

[0231] Unless defined otherwise, 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 any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, the 56NAI-5011166969vlpreferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited.

[0232] It must be noted that as used herein and in 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 optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as “solely,” “only” and the like in connection with the recitation of claim elements, or use of a “negative” limitation.

[0233] In some embodiments, the terms “first,” “second,” “third,” “fourth” and similar in a component name are used to distinguish and identify more than one component sharing certain identity in their names. For example, “first antibody” and “second antibody” are used to distinguish two antibodies.

[0234] It is understood that wherever embodiments are described herein with the term “comprising” otherwise analogous embodiments described in terms of “consisting of’ and / or “consisting essentially of’ are also provided. It is also understood that wherever embodiments are described herein with the phrase “consisting essentially of’ otherwise analogous embodiments described in terms of “consisting of’ are also provided.

[0235] The term “between” as used in a phrase as such “between A and B” or “between A-B” refers to a range including both A and B or any subranges thereof.

[0236] The term “and / or” as used in a phrase such as “A and / or B” herein is intended to include both A and B; A or B; A (alone); and B (alone). Likewise, the term “and / or” as used in a phrase such as “A, B, and / or C” is intended to encompass each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0237] The term “optional” or “optionally” means that the subsequently described circumstance may or may not occur, so that the description includes instances wherein the circumstance occurs, and the instances wherein the circumstance does not occur.

[0238] It is 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 sub-combination.57NAI-5011166969vl

[0239] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.TF-ADCS

[0240] An antibody that binds to TF (also referred to herein as “TF antibody,” “anti-TF antibody,” “TF Ab,” “Ab” or “antibody”) and a drug can be linked directly or indirectly to each other via a pyridazine-pyrrolo coupling moiety to form a TF-ADC as described herein. In certain embodiments, the TF antibody and the two or more drugs or active agents are bound to each other through 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.

[0241] Moi eties of interest (e.g., drugs or active agents) can be conjugated to the TF antibody at any desired site of the antibody. Thus, the present disclosure provides, for example, a TF antibody having moieties conjugated at two or more sites on the antibody, such as 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 the N-terminus of the antibody (e.g, at an internal site of the antibody). Combinations of the above conjugation sites are also possible.

[0242] In certain embodiments, a conjugate of the present disclosure includes two (or more) drugs or active agents conjugated to an amino acid residue of a TF antibody at the a-carbon of an amino acid residue. Stated another way, a conjugate includes a TF antibody where the side chain of an amino acid residue in the antibody has been modified and attached to two (or more) drugs or active agents (e.g, attached to two drugs or active agents through a branched linker as described herein). For example, a conjugate includes a TF antibody where the a-carbon of an amino acid residue in the antibody has been modified and attached to two drugs or active agents (e.g. , attached to two drugs or active agents through a branched linker as described herein).

[0243] Embodiments of the present disclosure include conjugates where a TF antibody is conjugated to two or more moieties, such as 3 moieties, 4 moieties, 5 moieties, 6 moieties, 7 moieties, 8 moieties, 9 moieties, 10 moieties, 11 moieties, 12 moieties, 13 moieties, 14 moieties, 15 moieties, 16 moieties, 17 moieties, 18 moieties, 19 moieties, or 20 or more58NAI-5011166969vlmoieties. The moieties may be conjugated to the TF antibody at multiple sites in the antibody. In some embodiments, two moieties may be conjugated to a single amino acid residue of the TF antibody. For instance, two moieties may be conjugated to the same amino acid residue of the TF antibody. In other embodiments, two moieties are conjugated to a first amino acid residue of the TF antibody and two other moieties are conjugated to a second amino acid residue of the TF antibody. For example, a TF antibody can be conjugated to first and second moieties at a first amino acid residue and conjugated to third and fourth moieties at a second amino acid residue, etc. In some cases, two or more amino acid residues in the TF antibody are each conjugated to a pair of moieties ( / .< ., two moieties), where each pair of moieties is conjugated to the TF antibody through a branched linker as described herein. In some cases, 1 amino acid residue in the TF antibody is conjugated to a pair of moieties through a branched linker as described herein. In other instances, 2 or more amino acid residues, such as 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 through a branched linker as described herein.

[0244] The one or more amino acid residues of the TF antibody that are conjugated to the moieties of interest may be naturally occurring amino acids, unnatural amino acids, or combinations thereof. For instance, the conjugate may include moieties of interest (e.g., drugs or active agents) conjugated to a naturally occurring amino acid residue of the TF antibody. In other instances, the conjugate may include moieties of interest conjugated to an unnatural amino acid residue of the TF antibody. The moieties of interest may be conjugated to the TF antibody at a single natural or unnatural amino acid residue as described above. One or more natural or unnatural amino acid residues in the TF antibody may be conjugated to the moieties of interest as described herein. For example, two (or more) amino acid residues (e.g., natural or unnatural amino acid residues) in the TF antibody may each be conjugated to two moieties through a branched linker, such that multiple sites in the TF antibody are conjugated to the moieties of interest.

[0245] As described herein, a TF antibody may be conjugated to two or more moieties of interest. In certain embodiments, the moiety of interest is a payload, for instance, a chemical entity, such as a drug, an active agent, or a detectable label. For example, drugs (or active agents, such as cytokines) may be conjugated to the TF antibody, or in other embodiments, detectable labels may be conjugated to the TF antibody. In other embodiments, combinations of different payloads may be conjugated to the TF antibody. Thus, for instance, embodiments of the present disclosure include, but are not limited to, the following: a conjugate of a TF antibody and two or more drugs; a conjugate of a TF antibody and two or more active agents,59NAI-5011166969vlsuch as cytokines; a conjugate of a TF antibody and two or more detectable labels; and combinations thereof.

[0246] In certain embodiments, the TF antibody and the moi eties of interest (e.g., drugs or active agents) are conjugated through a conjugation moiety. For example, the TF antibody and the moi eties of interest may each be bound (e.g., covalently bonded) to the conjugation moiety, thus indirectly binding the TF antibody and the moieties of interest together through the conjugation moiety. In some cases, the conjugation moiety includes a hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl compound, or a derivative of a hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl compound. For instance, a general scheme for coupling moieties of interest to a TF antibody through a hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety is shown in the general reaction scheme below. Hydrazinyl-indolyl and hydrazinyl-pyrrolo-pyridinyl conjugation moieties are also referred to herein as a hydrazino- / .w-Pictet-Spengler (HIPS) conjugation moiety and an aza-hy drazin o-iso- Pictet-Spengler (azaHIPS) conjugation moiety, respectively. Accordingly, in some embodiments, it would be understood by one of skill in the art that the carbon which is denoted in a formula as disclosed herein, such as Formula (I) or (II), as being immediately adjacent to Ab was originally a part of the antibody prior to the conjugation. In some embodiments, this carbon is conjugated to a (fGly’) residue, thus conjugating the antibody and the linker-payload. In other embodiments, this carbon is interpreted as a part of a (fGly’) residue conjugating the antibody and the linker-payload.

[0247] In the reaction scheme above, each R independently includes a moiety of interest (e.g., drug or active agent) that is conjugated to the TF antibody (e.g., conjugated to the TF antibody through a linker as described herein), where n’ is an integer from 1 to 4. As shown in the reaction scheme above, a conjugation moiety (e.g., a hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety) is attached to two or more drugs or active agents, R. A TF antibody that includes a 2-formylglycine residue (fGly) is reacted with the conjugation moiety to produce a TF antibody conjugate, thus attaching the two or more drugs or active agents to the TF antibody through the conjugation moiety.60NAI-5011166969vl

[0248] As described herein, the moieties can be any of a variety of moieties such as, but not limited to, chemical entities, such as detectable labels, or a drugs or active agents. R’ and R” may each independently be any desired substituent, such as, 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, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. Z may be CR21, NR22, N, O or S, where R21and R22are each independently selected from any of the substituents described for R’ and R” above.

[0249] Other hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moieties are also possible, as shown in the conjugates and compounds described herein. For example, the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moieties may be attached (e.g., covalently attached) to two or more linkers. As such, embodiments of the present disclosure include a hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety attached to two or more drugs or active agents each through a corresponding linker. Thus, conjugates of the present disclosure may include two or more linkers, where each linker attaches a corresponding drug or active agent to the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety. Accordingly, the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety and two or more linkers may be viewed overall as a “branched linker,” where the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety is attached to two of more “branches,” where each branch includes a linker attached to a drug or active agent.

[0250] Combinations of the same or different payloads may be conjugated to the TF antibody through the 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., drug, active agent or detectable label). For example, a first branch of a branched linker may be attached to a payload (e.g., drug, active agent or detectable label) and a second branch of the branched linker may be attached to the same payload (e.g., drug, active agent or detectable label) as the first branch.

[0251] 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 a branched linker may be attached to a first payload (e.g., a first drug, active agent or detectable label) and a second branch of the61NAI-5011166969vlbranched linker may be attached to a second payload (e.g., a second drug, active agent or detectable label) different from the first payload (e.g., the first drug, active agent or detectable label) attached to the first branch.

[0252] In some embodiments, where two different drugs or active agents are attached to the branched linker, the drugs or active agents may be selected from drugs and active agents that have a synergistic therapeutic effect. By “synergistic”, “synergism” or “synergy” is meant a therapeutic effect that is greater than the sum of the effects of the drugs or active agents taken separately. For example, in some instances, the use of two different drugs or active agents attached to the branched linker may provide a lower therapeutically effective concentration at which both payloads act, thereby increasing overall potency of the ADC.

[0253] In some embodiments, where two different drugs or active agents are attached to the branched linker, the drugs or active agents may be selected from drugs and active agents that provide an enhanced therapeutic benefit as compared to the use of the drugs or active agents separately. For example, the drugs or active agents may provide an increased effect on drug delivery of the ADC (e.g., some payloads, such as the iRGD peptide, can increase extravasation into tissues and augment tumor penetration).

[0254] In some embodiments, where two different drugs or active agents are attached to the branched linker, the drugs or active agents may be selected from drugs and active agents that use different mechanisms of action. In some cases, this may provide a decrease in tumor drug resistance by targeting multiple pathways. Examples of payload combinations can include, but are not limited to, cytotoxic drugs, immunomodulatory molecules to activate or inhibit immune cell populations, cytokines, hormones, chelating agents loaded with radioisotopes, and the like.

[0255] In some embodiments, where two different payloads are attached to the branched linker, the payloads may be selected from combinations of drugs or active agents and detectable labels. For example, a first payload may be a detectable label that is used as an imaging agent or tracer to detect the location of the ADC in vivo, while a second payload may be a drug or active agent that provides a therapeutic activity.

[0256] Various embodiments of the linkers that may couple the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety to the drugs or active agents are described in detail herein. For example, in some instances, the linker is a cleavable linker, such as a cleavable linker as described herein.

[0257] In certain embodiments, the TF antibody may be conjugated to two or more moieties of interest, where one or more amino acids of the TF antibody are modified before 62NAI-5011166969vlconjugation to the moieties of interest. Modification of one or more amino acids of the TF antibody may produce a TF antibody that contains one or more reactive groups suitable for conjugation to the moieties of interest. In some cases, the TF antibody may include one or more modified amino acid residues to provide one or more reactive groups suitable for conjugation to the moieties of interest (e.g., where two or more moieties are attached to a conjugation moiety, such as a hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety as described above). For example, an amino acid of the TF antibody may be modified to include a reactive aldehyde group (e.g., a reactive aldehyde). A reactive aldehyde may be included in an “aldehyde tag” or “aid-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) that has been converted by action of a formylglycine generating enzyme (FGE) to contain a 2-formylglycine residue (referred to herein as “fGly”). The fGly residue generated by an FGE may also be referred to as a “formylglycine”. Stated differently, the term “aldehyde tag” is used herein to refer to an amino acid sequence that includes a “converted” sulfatase motif ( / .< ., a sulfatase motif in which a cysteine or serine residue has been converted to fGly by action of an FGE, e.g., L(fGly)TPSR, SEQ ID NO: 123). A converted sulfatase motif may be produced from an amino acid sequence that includes an “unconverted” sulfatase motif ( / .< ., a sulfatase motif in which the cysteine or serine residue has not been converted to fGly by an FGE, but is capable of being converted, e.g., an unconverted sulfatase motif with the sequence: LCTPSR, SEQ ID NO: 100). By “conversion” as used in the context of action of a formylglycine generating enzyme (FGE) on a sulfatase motif refers to biochemical modification of a cysteine or serine residue in a sulfatase motif to a formylglycine (fGly) residue (e.g., Cys to fGly, or Ser to fGly). Additional aspects of aldehyde tags and uses thereof in site-specific protein modification are described in U.S. Patent No. 7,985,783 and U.S. Patent No. 8,729,232, the disclosures of each of which are incorporated herein by reference.

[0258] In some cases, to produce the conjugate, the TF antibody containing the fGly residue may be conjugated to the moieties of interest by reaction of the fGly with a compound (e.g., a compound containing a hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety, as described above). For example, an fGly-containing TF antibody may be contacted with a reactive partner under conditions suitable to provide for conjugation of two or more drugs to the TF antibody. In some instances, the reactive partner may include a hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety as described above. For example, two or more drugs or active agents may be attached to a 63NAI-5011166969vlhydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety. In some cases, the drugs or active agents are attached to a hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety, such as covalently attached to a hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl, where each drug or active agent is attached through a corresponding linker to the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety.

[0259] In certain embodiments, a conjugate of the present disclosure includes a TF antibody having at least one amino acid residue that has been attached to two or more moi eties of interest (e.g., drugs or active agents). In order to make the conjugate, an amino acid residue of the TF antibody may be modified and then coupled to two or more drugs or active agents attached to a hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety as described above. In certain embodiments, an amino acid residue of the TF antibody is a cysteine or serine residue that is modified to an fGly residue, as described above. In certain embodiments, the modified amino acid residue (e.g, fGly residue) is conjugated to two or more drugs or active agents containing a hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety as described above to provide a conjugate of the present disclosure where the two or more drugs or active agents are conjugated to the TF antibody through the hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety. As used herein, the term fGly’ refers to the amino acid residue of the TF antibody that is coupled to the moi eties of interest (e.g, drugs or active agents).

[0260] In certain embodiments, the conjugate includes a TF antibody having at least one amino acid residue attached to a branched linker as described herein, which in turn is attached to two or more drugs or active agents. For instance, the conjugate may include a TF antibody having at least one amino acid residue (fGly’) that is conjugated to the moieties of interest (e.g., drugs or active agents) as described above.

[0261] Aspects of the present disclosure include a conjugate of Formula (I):64NAI-5011166969vlwherein:Ab represents the antibody that binds to TF;Z1, Z2, Z3and Z4are each independently selected from CR4, N and C-LB-W2, wherein at least one Z1, Z2, Z3and Z4is C-LB-W2;R1is 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;R2and R3are 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, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, or R2and R3are optionally cyclically linked to form a 5 or 6-membered heterocyclyl;each R4is 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, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl;LAis a first linker;LBis a second linker;s is an integer from 1 to 10;W1is a first drug; andW2is a second drug.

[0262] In some embodiments, s is 2. In some embodiments, s is 4.

[0263] The substituents related to conjugates of Formula (I) are described in more detail below.

[0264] In certain embodiments, Z1, Z2, Z3and Z4are each independently selected from CR4, N and C-LB-W2, wherein at least one Z1, Z2, Z3and Z4is C-LB-W2. In certain embodiments, Z1is CR4. In certain embodiments, Z1is N. In certain embodiments, Z1is C-LB-W2. In certain embodiments, Z2is CR4. In certain embodiments, Z2is N. In certain embodiments, Z2is C-LB-W2. In certain embodiments, Z3is CR4. In certain embodiments, Z3is N. In certain embodiments, Z3is C-LB-W2. In certain embodiments, Z4is CR4. In certain 65NAI-5011166969vlembodiments, Z4is N. In certain embodiments, Z4is C-LB-W2. In some embodiments, each of Z1, Z3, and Z4is CR4. In some embodiments, Z3is C-LB-W2.

[0265] Combinations of various Z1, Z2, Z3and Z4are possible. For example, in some instances, Z1is C-LB-W2, Z2is CR4, Z3is CR4, and Z4is CR4. In some instances, Z1is CR4, Z2is C-LB-W2, Z3is CR4, and Z4is CR4. In some instances, Z1is CR4, Z2is CR4, Z3is C-LB-W2, and Z4is CR4. In some instances, Z1is CR4, Z2is CR4, Z3is CR4, and Z4is C-LB-W2.

[0266] In certain embodiments, R1is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, substituted heterocyclyl. In certain embodiments, R1is hydrogen. In certain embodiments, R1is alkyl or substituted alkyl, such as Ci-6 alkyl or Ci-6 substituted alkyl, or Ci-4 alkyl or Ci-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R1is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R1is alkynyl or substituted alkynyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R1is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a Cs aryl or Cs substituted aryl, or a Ce aryl or Ce substituted aryl. In certain embodiments, R1is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or C5-8 substituted heteroaryl, such as a Cs heteroaryl or Cs substituted heteroaryl, or a Ce heteroaryl or Ce substituted heteroaryl. In certain embodiments, R1is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R1is heterocyclyl or substituted heterocyclyl, such as C3-8 heterocyclyl or C3-8 substituted heterocyclyl, such as a C3-6 heterocyclyl or C3-6 substituted heterocyclyl, or a C3-5 heterocyclyl or C3-5 substituted heterocyclyl.

[0267] In certain embodiments, R2and R3are 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, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, or R2and R3are optionally cyclically linked to form a 5 or 6-membered heterocyclyl.66NAI-5011166969vl

[0268] In certain embodiments, R2is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R2is hydrogen. In certain embodiments, R2is alkyl or substituted alkyl, such as Ci-6 alkyl or Ci-6 substituted alkyl, or Ci-4 alkyl or Ci-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R2is methyl. In certain embodiments, R2is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R2is alkynyl or substituted alkynyl. In certain embodiments, R2is alkoxy or substituted alkoxy. In certain embodiments, R2is amino or substituted amino. In certain embodiments, R2is carboxyl or carboxyl ester. In certain embodiments, R2is acyl or acyloxy. In certain embodiments, R2is acyl amino or amino acyl. In certain embodiments, R2is alkylamide or substituted alkylamide. In certain embodiments, R2is sulfonyl. In certain embodiments, R2is thioalkoxy or substituted thioalkoxy. In certain embodiments, R2is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a Cs aryl or Cs substituted aryl, or a Ce aryl or Ce substituted aryl. In certain embodiments, R2is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or C5-8 substituted heteroaryl, such as a Cs heteroaryl or Cs substituted heteroaryl, or a Ce heteroaryl or Ce substituted heteroaryl. In certain embodiments, R2is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R2is heterocyclyl or substituted heterocyclyl, such as a C3-6 heterocyclyl or C3-6 substituted heterocyclyl, or a C3-5 heterocyclyl or C3-5 substituted heterocyclyl.

[0269] In certain embodiments, R3is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R3is hydrogen. In certain embodiments, R3is alkyl or substituted alkyl, such as C1-6 alkyl or C1-6 substituted alkyl, or Ci-4 alkyl or Ci-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R3is methyl. In certain embodiments, R3is alkenyl or substituted alkenyl, such 67NAI-5011166969vlas C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R3is alkynyl or substituted alkynyl. In certain embodiments, R3is alkoxy or substituted alkoxy. In certain embodiments, R3is amino or substituted amino. In certain embodiments, R3is carboxyl or carboxyl ester. In certain embodiments, R3is acyl or acyloxy. In certain embodiments, R3is acyl amino or amino acyl. In certain embodiments, R3is alkylamide or substituted alkylamide. In certain embodiments, R3is sulfonyl. In certain embodiments, R3is thioalkoxy or substituted thioalkoxy. In certain embodiments, R3is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a Cs aryl or Cs substituted aryl, or a Ce aryl or Ce substituted aryl. In certain embodiments, R3is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or C5-8 substituted heteroaryl, such as a Cs heteroaryl or Cs substituted heteroaryl, or a Ce heteroaryl or Ce substituted heteroaryl. In certain embodiments, R3is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R3is heterocyclyl or substituted heterocyclyl, such as C3-8 heterocyclyl or C3-8 substituted heterocyclyl, such as a C3-6 heterocyclyl or C3-6 substituted heterocyclyl, or a C3-5 heterocyclyl or C3-5 substituted heterocyclyl.

[0270] In certain embodiment, both R2and R3are methyl.

[0271] In certain embodiments, R2and R3are optionally cyclically linked to form a 5 or 6-membered heterocyclyl. In certain embodiments, R2and R3are cyclically linked to form a 5 or 6-membered heterocyclyl. In certain embodiments, R2and R3are cyclically linked to form a 5-membered heterocyclyl. In certain embodiments, R2and R3are cyclically linked to form a 6-membered heterocyclyl.

[0272] In certain embodiments, each R4is 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, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

[0273] The various possibilities for each R4are described in more detail as follows. In certain embodiments, R4is hydrogen. In certain embodiments, each R4is hydrogen. In certain embodiments, R4is halogen, such as F, Cl, Br or I. In certain embodiments, R4is F. In certain embodiments, R4is Cl. In certain embodiments, R4is Br. In certain embodiments, R4is I. In certain embodiments, R4is alkyl or substituted alkyl, such as C1-6 alkyl or C1-6 substituted 68NAI-5011166969vlalkyl, or Ci-4 alkyl or Ci-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R4is methyl. In certain embodiments, R4is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R4is alkynyl or substituted alkynyl. In certain embodiments, R4is alkoxy or substituted alkoxy. In certain embodiments, R4is amino or substituted amino. In certain embodiments, R4is carboxyl or carboxyl ester. In certain embodiments, R4is acyl or acyloxy. In certain embodiments, R4is acyl amino or amino acyl. In certain embodiments, R4is alkylamide or substituted alkylamide. In certain embodiments, R4is sulfonyl. In certain embodiments, R4is thioalkoxy or substituted thioalkoxy. In certain embodiments, R4is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a Cs aryl or Cs substituted aryl, or a Ce aryl or Ce substituted aryl (e.g., phenyl or substituted phenyl). In certain embodiments, R4is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or C5-8 substituted heteroaryl, such as a Cs heteroaryl or Cs substituted heteroaryl, or a Ce heteroaryl or Ce substituted heteroaryl. In certain embodiments, R4is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R4is heterocyclyl or substituted heterocyclyl, such as C3-8 heterocyclyl or C3-8 substituted heterocyclyl, such as a C3-6 heterocyclyl or C3-6 substituted heterocyclyl, or a C3-5 heterocyclyl or C3-5 substituted heterocyclyl.

[0274] In certain embodiments, LAis a first linker. Examples of linkers that can be used in the conjugates of the present disclosure are described in more detail below.

[0275] In certain embodiments, LBis a second linker. Examples of linkers that can be used in the conjugates of the present disclosure are described in more detail below.

[0276] In certain embodiments, W1is a first drug (or a first active agent). Examples of drugs and active agents that can be used in the conjugates of the present disclosure are described in more detail below.

[0277] In certain embodiments, W2is a second drug (or a second active agent). Examples of drugs and active agents that can be used in the conjugates of the present disclosure are described in more detail below.

[0278] In certain embodiments, Ab represents 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 rest of the conjugate69NAI-5011166969vlthrough an fGly’ residue as described herein. Examples of TF antibodies that can be used in the conjugates of the present disclosure are described in more detail below.

[0279] In certain embodiments, the conjugate of Formula (I) includes a first linker, LA. The first linker, LA, may be utilized to bind a first moiety of interest (e.g., a first drug or active agent) to a TF antibody through a conjugation moiety. The first linker, LA, may be bound (e.g., covalently bonded) to the conjugation moiety (e.g., as described herein). For example, the first linker, LA, may attach a hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety to a first drug. The hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety may be used to conjugate the first linker, LA, (and thus the first drug) to a TF antibody.

[0280] For example, as shown in Formula (I) above, LAis attached to Ab through a conjugation moiety, and thus Ab is indirectly bonded to the linker LAthrough the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety. As described above, Ab is a TF antibody, and thus LAis attached through the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety to the TF antibody, e.g., the linker LAis indirectly bonded to the TF antibody through the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety.

[0281] Any convenient linker may be utilized for the first linker LAin the subject conjugates and compounds. In certain embodiments, the first linker LAmay include a group selected from alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl amino, alkylamide, substituted alkylamide, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, the first linker LAmay include an alkyl or substituted alkyl group. In certain embodiments, the first linker LAmay include an alkenyl or substituted alkenyl group. In certain embodiments, the first linker LAmay include an alkynyl or substituted alkynyl group. In certain embodiments, the first linker LAmay include an alkoxy or substituted alkoxy group. In certain embodiments, the first linker LAmay include an amino or substituted amino group. In certain embodiments, the first linker LAmay include a carboxyl or carboxyl ester group. In certain embodiments, the first linker LAmay include an acyl amino group. In certain embodiments, the first linker LAmay include an alkylamide or substituted alkylamide group. In certain embodiments, the first linker LAmay include an aryl or substituted aryl group. In certain embodiments, the first linker LAmay include a heteroaryl or substituted heteroaryl group. In certain embodiments, the first linker LAmay include a cycloalkyl or 70NAI-5011166969vlsubstituted cycloalkyl group. In certain embodiments, the first linker LAmay include a heterocyclyl or substituted heterocyclyl group.

[0282] In certain embodiments, the first linker LAmay include a polymer. For example, the polymer may include a polyalkylene glycol and derivatives thereof, including polyethylene glycol, methoxypolyethylene glycol, polyethylene glycol homopolymers, polypropylene glycol homopolymers, copolymers of ethylene glycol with propylene glycol (e.g., where the homopolymers and copolymers are unsubstituted or substituted at one end with an alkyl group), polyvinyl alcohol, polyvinyl ethyl ethers, polyvinylpyrrolidone, combinations thereof, and the like. In certain embodiments, the polymer is a polyalkylene glycol. In certain embodiments, the polymer is a polyethylene glycol. Other linkers are also possible, as shown in the conjugates and compounds described in more detail below.

[0283] In some embodiments, LAis a first linker described by the formula:-(L^a-CL^b-CL^c-CL^d-CL^e-CL^f-,wherein L1, L2, L3, L4, L5and L6are each independently a linker subunit, and a, b, c, d, e and f are each independently 0 or 1, provided that at least one of a, b, c, d, e and f is 1.

[0284] 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.

[0285] In certain embodiments, the linker subunit L1is attached to the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety (e.g., as shown in Formula (I) above). In certain embodiments, the linker subunit L2, if present, is attached to the first drug or active agent W1. In certain embodiments, the linker subunit L3, if present, is attached to the first drug or active agent W1. In certain embodiments, the linker subunit L4, if present, is attached to the first drug or active agent W1. In certain embodiments, the linker subunit L5, if present, is attached to the first drug or active agent W1. In certain embodiments, the linker subunit L6, if present, is attached to the first drug or active agent W1.71NAI-5011166969vl

[0286] Any convenient linker subunits may be utilized in the first linker LA. Linker subunits of interest include, but are not limited to, units of polymers such as polyethylene glycols, polyethylenes and polyacrylates, amino acid residue(s), carbohydrate-based polymers or carbohydrate residues and derivatives thereof, polynucleotides, alkyl groups, aryl groups, heterocyclic groups, combinations thereof, and substituted versions thereof. In some embodiments, each of L1, L2, L3, L4, L5and L6(if present) comprises one or more groups independently selected from a polyethylene glycol, a modified polyethylene glycol, an amino acid residue, an alkyl group, a substituted alkyl, an aryl group, a substituted aryl group, and a diamine (e.g., a linking group that includes an alkylene diamine).

[0287] In some embodiments, L1(if present) comprises a polyethylene glycol, a 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, L1comprises a polyethylene glycol. In some embodiments, L1comprises a modified polyethylene glycol. In some embodiments, L1comprises an amino acid residue. In some embodiments, L1comprises an alkyl group or a substituted alkyl. In some embodiments, L1comprises an aryl group or a substituted aryl group. In some embodiments, L1comprises a diamine (e.g., a linking group comprising an alkylene diamine).

[0288] In some embodiments, L2(if present) comprises a polyethylene glycol, a 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, L2comprises a polyethylene glycol. In some embodiments, L2comprises a modified polyethylene glycol. In some embodiments, L2comprises an amino acid residue. In some embodiments, L2comprises an alkyl group or a substituted alkyl. In some embodiments, L2comprises an aryl group or a substituted aryl group. In some embodiments, L2comprises a diamine (e.g., a linking group comprising an alkylene diamine).

[0289] In some embodiments, L3(if present) comprises a polyethylene glycol, a 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, L3comprises a polyethylene glycol. In some embodiments, L3comprises a modified polyethylene glycol. In some embodiments, L3comprises an amino acid residue. In some embodiments, L3comprises an alkyl group or a substituted alkyl. In some embodiments, L3comprises an aryl group or a substituted aryl group. In some embodiments, L3comprises a diamine e.g., a linking group comprising an alkylene diamine).72NAI-5011166969vl

[0290] In some embodiments, L4(if present) comprises a polyethylene glycol, a 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, L4comprises a polyethylene glycol. In some embodiments, L4comprises a modified polyethylene glycol. In some embodiments, L4comprises an amino acid residue. In some embodiments, L4comprises an alkyl group or a substituted alkyl. In some embodiments, L4comprises an aryl group or a substituted aryl group. In some embodiments, L4comprises a diamine (e.g., a linking group comprising an alkylene diamine).

[0291] In some embodiments, L5(if present) comprises a polyethylene glycol, a 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, L5comprises a polyethylene glycol. In some embodiments, L5comprises a modified polyethylene glycol. In some embodiments, L5comprises an amino acid residue. In some embodiments, L5comprises an alkyl group or a substituted alkyl. In some embodiments, L5comprises an aryl group or a substituted aryl group. In some embodiments, L5comprises a diamine e.g., a linking group comprising an alkylene diamine).

[0292] In some embodiments, L6(if present) comprises a polyethylene glycol, a 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, L6comprises a polyethylene glycol. In some embodiments, L6comprises a modified polyethylene glycol. In some embodiments, L6comprises an amino acid residue. In some embodiments, L6comprises an alkyl group or a substituted alkyl. In some embodiments, L6comprises an aryl group or a substituted aryl group. In some embodiments, L6comprises a diamine (e.g., a linking group comprising an alkylene diamine).

[0293] In some embodiments, LAis a first linker comprising-(L1)a-(L2)b-(L3)c-(L4)d-(L5)e-(L6)f-, where:-(LJ)a- is -(TkvV;-(L2)b- is -(T2-V2)b-;-(L3)C- is -(T3-V3)C-;-(L4)d- is -(T4-V4)d-;-(L5)e- is -(T5-V5)e-; and-(L6)f- is -(T6-V6)f-,wherein T1, T2, T3, T4, T5and T6, if present, are tether groups;73NAI-5011166969vlV1, V2, V3, V4, V5and V6, if present, are covalent bonds or linking functional groups; anda, 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.

[0294] 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.

[0295] As described above, in certain embodiments, L1is attached to the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety (e.g., as shown in Formula (I) above). As such, in certain embodiments, T1is attached to the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety (e.g., as shown in Formula (I) above). In certain embodiments, V1is attached to the first drug or active agent. In certain embodiments, L2, if present, is attached to the first drug or active agent. As such, in certain embodiments, T2, if present, is attached to the first drug or active agent, or V2, if present, is attached to the first drug or active agent. In certain embodiments, L3, if present, is attached to the first drug or active agent. As such, in certain embodiments, T3, if present, is attached to the first drug or active agent, or V3, if present, is attached to the first drug or active agent. In certain embodiments, L4, if present, is attached to the first drug or active agent. As such, in certain embodiments, T4, if present, is attached to the first drug or active agent, or V4, if present, is attached to the first drug or active agent. In certain embodiments, L5, if present, is attached to the first drug or active agent. As such, in certain embodiments, T5, if present, is attached to the first drug or active agent, or V5, if present, is attached to the first drug or active agent. In certain embodiments, L6, if present, is attached to the first drug or active agent. As such, in certain embodiments, T6, if present, is attached to the first drug or active agent, or V6, if present, is attached to the first drug or active agent.

[0296] In certain embodiments, the conjugate of Formula (I) includes a second linker, LB. The second linker, LB, may be utilized to bind a second moiety of interest (e.g., a second drug 74NAI-5011166969vlor active agent) to a TF antibody through a conjugation moiety. The second linker, LB, may be bound (e.g., covalently bonded) to the conjugation moiety (e.g., as described herein). For example, the second linker, LB, may attach a hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety to a second drug. The hydrazinyl-indolyl or hydrazinyl-pyrrolo-pyridinyl conjugation moiety may be used to conjugate the second linker, LB, (and thus the second drug) to a TF antibody.

[0297] For example, as shown in Formula (I) above, LBis attached to Ab through a conjugation moiety, and thus Ab is indirectly bonded to the second linker LBthrough the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety. As described above, Ab is a TF antibody, and thus LBis attached through the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety to the TF antibody, e.g., the linker LBis indirectly bonded to the TF antibody through the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety.

[0298] Any convenient linker may be utilized for the second linker LBin the subject conjugates and compounds. In certain embodiments, the second linker LBmay include a group selected from alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl amino, alkylamide, substituted alkylamide, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, the second linker LBmay include an alkyl or substituted alkyl group. In certain embodiments, the second linker LBmay include an alkenyl or substituted alkenyl group. In certain embodiments, the second linker LBmay include an alkynyl or substituted alkynyl group. In certain embodiments, the second linker LBmay include an alkoxy or substituted alkoxy group. In certain embodiments, the second linker LBmay include an amino or substituted amino group. In certain embodiments, the second linker LBmay include a carboxyl or carboxyl ester group. In certain embodiments, the second linker LBmay include an acyl amino group. In certain embodiments, the second linker LBmay include an alkylamide or substituted alkylamide group. In certain embodiments, the second linker LBmay include an aryl or substituted aryl group. In certain embodiments, the second linker LBmay include a heteroaryl or substituted heteroaryl group. In certain embodiments, the second linker LBmay include a cycloalkyl or substituted cycloalkyl group. In certain embodiments, the second linker LBmay include a heterocyclyl or substituted heterocyclyl group.

[0299] In certain embodiments, the second linker LBmay include a polymer. For example, the polymer may include a polyalkylene glycol and derivatives thereof, including75NAI-5011166969vlpolyethylene glycol, methoxypolyethylene glycol, polyethylene glycol homopolymers, polypropylene glycol homopolymers, copolymers of ethylene glycol with propylene glycol (e.g., where the homopolymers and copolymers are unsubstituted or substituted at one end with an alkyl group), polyvinyl alcohol, polyvinyl ethyl ethers, polyvinylpyrrolidone, combinations thereof, and the like. In certain embodiments, the polymer is a polyalkylene glycol. In certain embodiments, the polymer is a polyethylene glycol. Other linkers are also possible, as shown in the conjugates and compounds described in more detail below.

[0300] In some embodiments, LBis a second linker described by the formula:wherein L7, L8, L9, L10, L11, L12and L13are each independently a linker subunit, and g, h, i, j, k, 1 and m are each independently 0 or 1, provided at least one of g, h, i, j, k, 1 and m is 1.

[0301] In certain embodiments, the sum of g, h, i, j, k, 1 and m is 1 to 7. In certain embodiments, the sum of g, h, i, j, k, 1 and m is 1. In certain embodiments, the sum of g, h, i, j, k, 1 and m is 2. In certain embodiments, the sum of g, h, i, j, k, 1 and m is 3. In certain embodiments, the sum of g, h, i, j, k, 1 and m is 4. In certain embodiments, the sum of g, h, i, j, k, 1 and m is 5. In certain embodiments, the sum of g, h, i, j, k, 1 and m is 6. In certain embodiments, the sum of g, h, i, j, k, 1 and m is 7. In certain embodiments, g, h, i, j, k, 1 and m are each 1. In certain embodiments, g, h, i, j, k and 1 are each 1 and m is 0. In certain embodiments, g, h, i, j and k are each 1 and 1 and m are each 0. In certain embodiments, g, h, i and j are each 1 and k, 1 and m are each 0. In certain embodiments, g, h, and i are each 1 and j, k, 1 and m are each 0. In certain embodiments, g and h are each 1 and i, j, k, 1 and m are each 0. In certain embodiments, g is 1 and h, i, j, k, 1 and m are each 0. In certain embodiments, g, h, i, j, k, 1 and m are each 0.

[0302] In certain embodiments, the linker subunit L7is attached to the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety (e.g., as shown in Formula (I) above). In certain embodiments, the linker subunit L8, if present, is attached to the second drug or active agent W2. In certain embodiments, the linker subunit L9, if present, is attached to the second drug or active agent W2. In certain embodiments, the linker subunit L10, if present, is attached to the second drug or active agent W2. In certain embodiments, the linker subunit L11, if present, is attached to the second drug or active agent W2. In certain embodiments, the linker subunit L12, if present, is attached to the second drug or active agent W2. In certain embodiments, the linker subunit L13, if present, is attached to the second drug or active agent W2.76NAI-5011166969vl

[0303] Any convenient linker subunits may be utilized in the second linker LB. Linker subunits of interest include, but are not limited to, units of polymers such as polyethylene glycols, polyethylenes and polyacrylates, amino acid residue(s), carbohydrate-based polymers or carbohydrate residues and derivatives thereof, polynucleotides, alkyl groups, aryl groups, heterocyclic groups, combinations thereof, and substituted versions thereof. In some embodiments, each of L7, L8, L9, L10, L11, L12and L13(if present) comprise one or more groups independently selected from a polyethylene glycol, a modified polyethylene glycol, an amino acid residue, an alkyl group, a substituted alkyl, an aryl group, a substituted aryl group, and a diamine (e.g., a linking group that includes an alkylene diamine).

[0304] In some embodiments, L7(if present) comprises a polyethylene glycol, a 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, L7comprises a polyethylene glycol. In some embodiments, L7comprises a modified polyethylene glycol. In some embodiments, L7comprises an amino acid residue. In some embodiments, L7comprises an alkyl group or a substituted alkyl. In some embodiments, L7comprises an aryl group or a substituted aryl group. In some embodiments, L7comprises a diamine (e.g., a linking group comprising an alkylene diamine).

[0305] In some embodiments, L8(if present) comprises a polyethylene glycol, a 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, L8comprises a polyethylene glycol. In some embodiments, L8comprises a modified polyethylene glycol. In some embodiments, L8comprises an amino acid residue. In some embodiments, L8comprises an alkyl group or a substituted alkyl. In some embodiments, L8comprises an aryl group or a substituted aryl group. In some embodiments, L8comprises a diamine (e.g., a linking group comprising an alkylene diamine).

[0306] In some embodiments, L9(if present) comprises a polyethylene glycol, a 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, L9comprises a polyethylene glycol. In some embodiments, L9comprises a modified polyethylene glycol. In some embodiments, L9comprises an amino acid residue. In some embodiments, L9comprises an alkyl group or a substituted alkyl. In some embodiments, L9comprises an aryl group or a substituted aryl group. In some embodiments, L9comprises a diamine (e.g., a linking group comprising an alkylene diamine).77NAI-5011166969vl

[0307] In some embodiments, L10(if present) comprises a polyethylene glycol, a 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, L10comprises a polyethylene glycol. In some embodiments, L10comprises a modified polyethylene glycol. In some embodiments, L10comprises an amino acid residue. In some embodiments, L10comprises an alkyl group or a substituted alkyl. In some embodiments, L10comprises an aryl group or a substituted aryl group. In some embodiments, L10comprises a diamine (e.g., a linking group comprising an alkylene diamine).

[0308] In some embodiments, L11(if present) comprises a polyethylene glycol, a 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, L11comprises a polyethylene glycol. In some embodiments, L11comprises a modified polyethylene glycol. In some embodiments, L11comprises an amino acid residue. In some embodiments, L11comprises an alkyl group or a substituted alkyl. In some embodiments, L11comprises an aryl group or a substituted aryl group. In some embodiments, L11comprises a diamine (e.g., a linking group comprising an alkylene diamine).

[0309] In some embodiments, L12(if present) comprises a polyethylene glycol, a 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, L12comprises a polyethylene glycol. In some embodiments, L12comprises a modified polyethylene glycol. In some embodiments, L12comprises an amino acid residue. In some embodiments, L12comprises an alkyl group or a substituted alkyl. In some embodiments, L12comprises an aryl group or a substituted aryl group. In some embodiments, L12comprises a diamine (e.g., a linking group comprising an alkylene diamine).

[0310] In some embodiments, L13(if present) comprises a polyethylene glycol, a 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, L13comprises a polyethylene glycol. In some embodiments, L13comprises a modified polyethylene glycol. In some embodiments, L13comprises an amino acid residue. In some embodiments, L13comprises an alkyl group or a substituted alkyl. In some embodiments, L13comprises an aryl group or a substituted aryl group. In some embodiments, L13comprises a diamine e.g., a linking group comprising an alkylene diamine).

[0311] In some embodiments, LBis a second linker comprising-(^-(L8^-^9);-^10^-^11^-^12)!-^13^-, where:78NAI-5011166969vl-(L7)g- is -(T7-V7)g-;-(L8)h- is -(T8-V8)h-;-(L9)i- is -(T9-V9)i-;-(L10)j- is -(T10-V10)j-;-(L11)k- is -(T11-V11)k-;-(L12)I- is -(T12-V12)I-; and-(L13)m- is -(T13-V13)m-,wherein T7, T8, T9, T10, T11, T12and T13, if present, are tether groups;V7, V8, V9, V10, V11, V12and V13, if present, are covalent bonds or linking functional groups; andg, h, i, j, k, 1 and m are each independently 0 or 1, provided at least one of g, h, i, j, k, 1, and m is 1.

[0312] In certain embodiments, the sum of g, h, i, j, k, 1 and m is 1 to 7. In certain embodiments, the sum of g, h, i, j, k, 1 and m is 1. In certain embodiments, the sum of g, h, i, j, k, 1 and m is 2. In certain embodiments, the sum of g, h, i, j, k, 1 and m is 3. In certain embodiments, the sum of g, h, i, j, k, 1 and m is 4. In certain embodiments, the sum of g, h, i, j, k, 1 and m is 5. In certain embodiments, the sum of g, h, i, j, k, 1 and m is 6. In certain embodiments, the sum of g, h, i, j, k, 1 and m is 7. In certain embodiments, g, h, i, j, k, 1 and m are each 1. In certain embodiments, g, h, i, j, k and 1 are each 1 and m is 0. In certain embodiments, g, h, i, j and k are each 1 and 1 and m are each 0. In certain embodiments, g, h, i and j are each 1 and k, 1 and m are each 0. In certain embodiments, g, h, and i are each 1 and j, k, 1 and m are each 0. In certain embodiments, g and h are each 1 and i, j, k, 1 and m are each 0. In certain embodiments, g is 1 and h, i, j, k, 1 and m are each 0. In certain embodiments, g, h, i, j, k, 1 and m are each 0.

[0313] As described above, in certain embodiments, L7is attached to the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety (e.g., as shown in Formula (I) above). As such, in certain embodiments, T7is attached to the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety (e.g., as shown in Formula (I) above). In certain embodiments, V7is attached to the second drug or active agent. In certain embodiments, L8, if present, is attached to the second drug or active agent. As such, in certain embodiments, T8, if present, is attached to the second drug or active agent, or V8, if present, is attached to the second drug or active agent. In certain embodiments, L9, if present, is attached to the second drug or active agent. As such, in certain embodiments, T9, if present, is attached to the second drug or active agent, or V9, if present, is attached to the second drug or active agent. In 79NAI-5011166969vlcertain embodiments, L10, if present, is attached to the second drug or active agent. As such, in certain embodiments, T10, if present, is attached to the second drug or active agent, or VI 04, if present, is attached to the second drug or active agent. In certain embodiments, L11, if present, is attached to the second drug or active agent. As such, in certain embodiments, T11, if present, is attached to the second drug or active agent, or V11, if present, is attached to the second drug or active agent. In certain embodiments, L12, if present, is attached to the second drug or active agent. As such, in certain embodiments, T12, if present, is attached to the second drug or active agent, or V12, if present, is attached to the second drug or active agent. In certain embodiments, L13, if present, is attached to the second drug or active agent. As such, in certain embodiments, T13, if present, is attached to the second drug or active agent, or V13, if present, is attached to the second drug or active agent.

[0314] Regarding the tether groups, T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and T13, any convenient tether groups may be utilized in the subject linkers. In some embodiments, T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and T13each comprise one or more groups independently selected from a covalent bond, a (Ci-Cnjalkyl, a substituted (Ci-Cnjalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA)W, (PEG)n, (AA)P, -(CR13OH)X-, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), paraaminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), parahydroxy-phenyl (PHP), an acetal group, a hydrazine, a disulfide, and an ester, where each w is an integer from 1 to 20, each n is an integer from 1 to 30, each p is an integer from 1 to 20, and each x is an integer from 1 to 12.

[0315] In certain embodiments, the tether group e.g., T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and / or T13) includes a (Ci-Ci2)alkyl or a substituted (Ci-Ci2)alkyl. In certain embodiments, (Ci-Ci2)alkyl is a straight chain or branched alkyl group that includes from 1 to 12 carbon atoms, such as 1 to 10 carbon atoms, or 1 to 8 carbon atoms, or 1 to 6 carbon atoms, or 1 to 5 carbon atoms, or 1 to 4 carbon atoms, or 1 to 3 carbon atoms. In some instances, (Ci-Ci2)alkyl may be an alkyl or substituted alkyl, such as C1-C12 alkyl, or C1-C10 alkyl, or Ci-Ce alkyl, or C1-C3 alkyl. In some instances, (Ci-Ci2)alkyl is a C2-alkyl. For example, (Ci-Ci2)alkyl may be an alkylene or substituted alkylene, such as C1-C12 alkylene, or C1-C10 alkylene, or Ci-Ce alkylene, or C1-C3 alkylene. In some instances, (Ci-Ci2)alkyl is a Ci-alkylene (e.g., CH2). In some instances, (Ci-Ci2)alkyl is a C2-alkylene (e.g., CH2CH2). In some instances, (Ci-Ci2)alkyl is a C3-alkylene (e.g., CH2CH2CH2).80NAI-5011166969vl

[0316] In certain embodiments, substituted (Ci-Ci2)alkyl is a straight chain or branched substituted alkyl group that includes from 1 to 12 carbon atoms, such as 1 to 10 carbon atoms, or 1 to 8 carbon atoms, or 1 to 6 carbon atoms, or 1 to 5 carbon atoms, or 1 to 4 carbon atoms, or 1 to 3 carbon atoms. In some instances, substituted (Ci-Ci2)alkyl may be a substituted alkyl, such as substituted C1-C12 alkyl, or substituted C1-C10 alkyl, or substituted Ci-Ce alkyl, or substituted C1-C3 alkyl. In some instances, substituted (Ci-Ci2)alkyl is a substituted C2-alkyl. For example, substituted (Ci-Ci2)alkyl may be a substituted alkylene, such as substituted C1-C12 alkylene, or substituted C1-C10 alkylene, or substituted Ci-Ce alkylene, or substituted C1-C3 alkylene. In some instances, substituted (Ci-Ci2)alkyl is a substituted Ci-alkylene (e.g., Ci-alkylene substituted with -SO3H). In some instances, substituted (Ci-Ci2)alkyl is a substituted C2-alkylene. In some instances, substituted (Ci-Cn)alkyl is a substituted C3-alkylene. For example, substituted (Ci-Ci2)alkyl may include Ci-C12 alkylene (e.g., C3-alkylene or Cs-alkylene) substituted with a (PEG)k’ group as described herein (e.g,-CONH(PEG>, such as -CONH(PEG)3or -CONH(PEG)5; or -NHCO(PEG)k’, such as -NHCO(PEG)?), or may include C1-C12 alkylene (e.g., C3-alkylene) substituted with a -CONHCH2CH2SO3H group, or may include C1-C12 alkylene (e.g., Cs-alkylene) substituted with a -NHCOCH2SO3H group.

[0317] In certain embodiments, the tether group (e.g., T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and / or T13) includes an aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, or substituted heterocyclyl. In some instances, the tether group (e.g., T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and / or T13) includes an aryl or substituted aryl. For example, the aryl can be phenyl. In some cases, the substituted aryl is a substituted phenyl. The substituted phenyl can be substituted with one or more substituents selected from (Ci-Ci2)alkyl, a substituted (Ci-Ci2)alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In some instances, the substituted aryl is a substituted phenyl, where the substituent includes a cleavable moiety as described herein (e.g, an enzymatically cleavable moiety, such as a glycoside or glycoside derivative).

[0318] In some instances, the tether group (e.g, T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and / or T13) includes a heteroaryl or substituted heteroaryl, such triazolyl (e.g., 1,2,3-triazolyl). In some instances, the tether group (e.g., T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and / or T13) includes a cycloalkyl or substituted cycloalkyl. In some instances, the tether group (e.g., T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and / or T13) includes a heterocyclyl or substituted heterocyclyl. In some instances, the substituent on the substituted heteroaryl,81NAI-5011166969vlsubstituted cycloalkyl or substituted heterocyclyl includes a cleavable moiety as described herein (e.g., an enzymatically cleavable moiety, such as a glycoside or glycoside derivative).

[0319] In certain embodiments, the tether group (e.g., T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and / or T13) includes an ethylene diamine (EDA) moiety, e.g, an EDA containing tether group. In certain embodiments, (EDA)Wincludes one or more EDA moieties, such as where w is an integer from 1 to 50, such as from 1 to 40, from 1 to 30, from 1 to 20, from 1 to 12 or from 1 to 6, such as 1, 2, 3, 4, 5 or 6). The linked ethylene diamine (EDA) moieties may optionally be substituted at one or more convenient positions with any convenient substituents, e.g, with an alkyl, a substituted alkyl, an acyl, a substituted acyl, an aryl or a substituted aryl. In certain embodiments, the EDA moiety is described by the structure:where y is an integer from 1 to 6, r is 0 or 1, and each R12is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, y is 1, 2, 3, 4, 5 or 6. In certain embodiments, y is 1 and r is 0. In certain embodiments, y is 1 and r is 1. In certain embodiments, y is 2 and r is 0. In certain embodiments, y is 2 and r is 1. In certain embodiments, each R12is independently selected from hydrogen, an alkyl, a substituted alkyl, an aryl and a substituted aryl. In certain embodiments, any two adjacent R12groups of the EDA may be cyclically linked, e.g., to form a piperazinyl ring. In certain embodiments, y is 1 and the two adjacent R12groups are an alkyl group, cyclically linked to form a piperazinyl ring. In certain embodiments, y is 1 and the adjacent R12groups are selected from hydrogen, an alkyl (e.g, methyl) and a substituted alkyl (e.g, lower alkyl-OH, such as ethyl-OH or propyl-OH).

[0320] In certain embodiments, the tether group (e.g., T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and / or T13) includes a 4-amino-piperidine (4AP) moiety (also referred to herein as piperidin-4-amino, P4A). The 4AP moiety may optionally be substituted at one or more convenient positions with any convenient substituents, e.g., with an alkyl, a substituted alkyl,82NAI-5011166969vla polyethylene glycol moiety, an acyl, a substituted acyl, an aryl or a substituted aryl. In certain embodiments, the 4AP moiety is described by the structure:where R12is selected from hydrogen, alkyl, substituted alkyl, a polyethylene glycol moiety (e.g., a polyethylene glycol or a modified polyethylene glycol), alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R12is a polyethylene glycol moiety. In certain embodiments, R12is a carboxy modified polyethylene glycol.

[0321] In certain embodiments, R12includes a polyethylene glycol moiety described by the formula: (PEG)k’, which may be represented by the structure:where k’ is an integer from 1 to 20, such as from 1 to 18, or from 1 to 16, or from 1 to 14, or from 1 to 12, or from 1 to 10, or from 1 to 8, or from 1 to 6, or from 1 to 4, or 1 or 2, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20. In some instances, k’ is 2. In certain embodiments, R17is selected from OH, COOH, OR, or COOR, where R is selected from alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R17is COOH. In certain embodiments, R17is OH. In certain embodiments, R17is OCH3.

[0322] In certain embodiments, a tether group (e.g., T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and / or T13) includes (PEG)n, where (PEG)n is a polyethylene glycol or a modified polyethylene glycol linking unit. In certain embodiments, (PEG)n is described by the structure:where n is an integer from 1 to 50, such as from 1 to 40, from 1 to 30, from 1 to 20, from 1 to 12 or from I to 6, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20.83NAI-5011166969vlIn some instances, n is 2. In some instances, n is 3. In some instances, n is 6. In some instances, n is 12.

[0323] In certain embodiments, a tether group (e.g., T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and / or T13) includes (AA)P, where AA is an amino acid residue. Any convenient amino acids may be utilized. Amino acids of interest include but are not limited to, L- and D-amino acids, naturally occurring amino acids such as any of the 20 primary alpha-amino acids and beta-alanine, non-naturally occurring amino acids (e.g., amino acid analogs), such as a non-naturally occurring alpha-amino acid or a non-naturally occurring beta-amino acid, etc. In certain embodiments, p is an integer from 1 to 50, such as from 1 to 40, from 1 to 30, from 1 to 20, from 1 to 12 or from 1 to 6, such as 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.

[0324] In certain embodiments, a tether group (e.g., T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and / or T13) includes an amino acid analog. Amino acid analogs include compounds that are similar in structure and / or overall shape to one or more amino acids commonly found in naturally occurring proteins e.g., Ala or A, Cys or C, Asp or D, Glu or E, Phe or F, Gly or G, His or H, He or I, Lys or K, Leu or L, Met or M, Asn or N, Pro or P, Gin or Q, Arg or R, Ser or S, Thr or T, Vai 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 with the same stereochemistry as in the naturally occurring D-form, as well as the L-form of amino acid analogs. In some instances, the amino acid analogs share backbone structures, and / or the side chain structures of one or more natural amino acids, with difference(s) being one or more modified groups in the molecule. Such modification may include, but is not limited to, substitution of an atom (such as N) for a related atom (such as S), addition of a group (such as methyl, or hydroxyl, etc.) or an atom (such as Cl or Br, etc.), deletion of a group, substitution of a covalent bond (single bond for double bond, etc.), or combinations thereof. For example, amino acid analogs may include a-hydroxy acids, and a-amino acids, and the like. Examples of amino acid analogs include, but are not limited to, sulfoalanine, and the like.

[0325] In certain embodiments, a tether group (e.g., T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and / or T13) includes a moiety described by the formula -(CR13OH)X-, where x is 0 or x is an integer from 1 to 50, such as from 1 to 40, from 1 to 30, from 1 to 20, from 1 to 12 or from 1 to 6, such as 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, R13is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy,84NAI-5011166969vlsubstituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl. In certain embodiments, R13is hydrogen. In certain embodiments, R13is alkyl or substituted alkyl, such as Ci-6 alkyl or Ci-6 substituted alkyl, or Ci-4 alkyl or Ci-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R13is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R13is alkynyl or substituted alkynyl. In certain embodiments, R13is alkoxy or substituted alkoxy. In certain embodiments, R13is amino or substituted amino. In certain embodiments, R13is carboxyl or carboxyl ester. In certain embodiments, R13is acyl or acyloxy. In certain embodiments, R13is acyl amino or amino acyl. In certain embodiments, R13is alkylamide or substituted alkylamide. In certain embodiments, R13is sulfonyl. In certain embodiments, R13is thioalkoxy or substituted thioalkoxy. In certain embodiments, R13is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a Cs aryl or Cs substituted aryl, or a Ce aryl or Ce substituted aryl. In certain embodiments, R13is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or Cs-8 substituted heteroaryl, such as a Cs heteroaryl or Cs substituted heteroaryl, or a Ce heteroaryl or Ce substituted heteroaryl. In certain embodiments, R13is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R13is heterocyclyl or substituted heterocyclyl, such as C3-8 heterocyclyl or C3-8 substituted heterocyclyl, such as a C3-6 heterocyclyl or C3-6 substituted heterocyclyl, or a C3-5 heterocyclyl or C3-5 substituted heterocyclyl.

[0326] In certain embodiments, R13is selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. In these embodiments, alkyl, substituted alkyl, aryl, and substituted aryl are as described above for R13.

[0327] In certain embodiments, the tether group (e.g., T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and / or T13) includes an acetal group, a disulfide, a hydrazine, or an ester. In some embodiments, the tether group includes an acetal group. In some embodiments, the tether group includes a hydrazine. In some embodiments, the tether group includes a disulfide. In some embodiments, the tether group includes an ester.

[0328] In certain embodiments, a tether group (e.g., T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and / or T13) includes a meta-amino-benzyloxy (MABO), meta-amino- 85NAI-5011166969vlbenzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), or para-hydroxy -phenyl (PHP).

[0329] In some embodiments, a tether group includes a MABO group described by the following structure:

[0330] In some embodiments, a tether group includes a MABC group described by the following structure:

[0331] In some embodiments, a tether group includes a PABO group described by the following structure:R14

[0332] In some embodiments, a tether group includes a PABC group described by the following structure:

[0333] In some embodiments, a tether group includes a PAB group described by the following structure:

[0334] In some embodiments, a tether group includes a PABA group described by the following structure:86NAI-5011166969vlR14

[0335] In some embodiments, a tether group includes a PAP group described by the following structure:R14

[0336] In some embodiments, a tether group includes a PHP group described by the following structure:

[0337] In certain embodiments, each R14is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

[0338] In certain embodiments, R14is hydrogen. In certain embodiments, each R14is hydrogen. In certain embodiments, R14is alkyl or substituted alkyl, such as Ci-6 alkyl or Ci-6 substituted alkyl, or Ci-4 alkyl or Ci-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R14is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R14is alkynyl or substituted alkynyl. In certain embodiments, R14is alkoxy or substituted alkoxy. In certain embodiments, R14is amino or substituted amino. In certain embodiments, R14is carboxyl or carboxyl ester. In certain embodiments, R14is acyl or acyloxy. In certain embodiments, R14is acyl amino or amino acyl. In certain embodiments, R14is alkylamide or substituted alkylamide. In certain embodiments, R14is sulfonyl. In certain embodiments, R14is thioalkoxy or substituted thioalkoxy. In certain embodiments, R14is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a Cs aryl or Cs substituted aryl, or a Ce aryl or Ce substituted aryl. In certain embodiments, R14is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or Cs-87NAI-5011166969vl8 substituted heteroaryl, such as a Cs heteroaryl or Cs substituted heteroaryl, or a Ce heteroaryl or Ce substituted heteroaryl. In certain embodiments, R14is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R14is heterocyclyl or substituted heterocyclyl, such as C3-8 heterocyclyl or C3-8 substituted heterocyclyl, such as a C3-6 heterocyclyl or C3-6 substituted heterocyclyl, or a C3-5 heterocyclyl or C3-5 substituted heterocyclyl.

[0339] In some embodiments of the MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP tether structures shown above, the phenyl ring may be substituted with one or more additional groups selected from halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

[0340] In certain embodiments, one or more of the tether groups T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12and / or T13is each optionally substituted with a glycoside or glycoside derivative. For example, in some instances, T1, T2, T3, T4, T5and T6are each optionally substituted with a glycoside. In some instances, T7, T8, T9, T10, T11, T12and T13are each optionally substituted with a glycoside. In certain embodiments, the glycoside or glycoside derivative is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.

[0341] In certain embodiments, the MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP tether structures shown above may be substituted with one or more additional groups selected from a glycoside and a glycoside derivative. For example, in some embodiments of the MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP tether structures shown above, the phenyl ring may be substituted with one or more additional groups selected from a glycoside and a glycoside derivative. In certain embodiments, the glycoside or glycoside derivative is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc. In some embodiments, the PABC is substituted with a glycoside, for example, a hydrogen of PABC is replaced with a glycoside, such as a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.

[0342] For example, in some embodiments, the glycoside or glycoside derivative can be selected from the following structures:88NAI-5011166969vlOH O OH QH OH

[0343] Regarding the linking functional groups, V1, V2, V3, V4, V5, V6, V7, V8, V9, V10, V11, V12and V13any convenient linking functional groups may be utilized in the subject linkers. Linking functional groups of interest include, but are not limited to, amino, carbonyl, amido, oxycarbonyl, carboxy, sulfonyl, sulfoxide, sulfonylamino, aminosulfonyl, thio, oxy, phospho, phosphoramidate, thiophosphoraidate, and the like. In some embodiments, V1, V2, V3, V4, V5, V6, V7, V8, V9, V10, V11, V12and V13are each independently selected from a covalent bond, -CO-, -NR15-, -NR15(CH2)q-, -NR15(C6H4)-, -CONR15-, -NR15CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR15-, -NR15SO2- 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.

[0344] In some embodiments, each R15is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.

[0345] In certain embodiments, R15is hydrogen. In certain embodiments, each R15is hydrogen. In certain embodiments, R15is alkyl or substituted alkyl, such as C1-6 alkyl or C1-6 substituted alkyl, or Ci-4 alkyl or Ci-4 substituted alkyl, or C1-3 alkyl or C1-3 substituted alkyl. In certain embodiments, R15is alkenyl or substituted alkenyl, such as C2-6 alkenyl or C2-6 substituted alkenyl, or C2-4 alkenyl or C2-4 substituted alkenyl, or C2-3 alkenyl or C2-3 substituted alkenyl. In certain embodiments, R15is alkynyl or substituted alkynyl. In certain embodiments, R15is alkoxy or substituted alkoxy. In certain embodiments, R15is amino or substituted amino. In certain embodiments, R15is carboxyl or carboxyl ester. In certain 89NAI-5011166969vlembodiments, R15is acyl or acyloxy. In certain embodiments, R15is acyl amino or amino acyl. In certain embodiments, R15is alkylamide or substituted alkylamide. In certain embodiments, R15is sulfonyl. In certain embodiments, R15is thioalkoxy or substituted thioalkoxy. In certain embodiments, R15is aryl or substituted aryl, such as C5-8 aryl or C5-8 substituted aryl, such as a C5 aryl or C5 substituted aryl, or a Ce aryl or Ce substituted aryl. In certain embodiments, R15is heteroaryl or substituted heteroaryl, such as C5-8 heteroaryl or C5-8 substituted heteroaryl, such as a C5 heteroaryl or C5 substituted heteroaryl, or a Ce heteroaryl or Ce substituted heteroaryl. In certain embodiments, R15is cycloalkyl or substituted cycloalkyl, such as C3-8 cycloalkyl or C3-8 substituted cycloalkyl, such as a C3-6 cycloalkyl or C3-6 substituted cycloalkyl, or a C3-5 cycloalkyl or C3-5 substituted cycloalkyl. In certain embodiments, R15is heterocyclyl or substituted heterocyclyl, such as C3-8 heterocyclyl or C3-8 substituted heterocyclyl, such as a C3-6 heterocyclyl or C3-6 substituted heterocyclyl, or a C3-5 heterocyclyl or C3-5 substituted heterocyclyl.

[0346] In certain embodiments, each R15is 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 as described above for R15.

[0347] As described above, in some embodiments, LAis a first linker comprising -(TkV^a-(T2-V2)b-(T3-V3)c-(T4-V4)d-(T5-V5)e-(T6-V6)f-, where a, b, c, d, e and f are each independently 0 or 1, provided at least one of a, b, c, d, e, and f is 1.

[0348] In some embodiments, in the first linker LA:T1is selected from a (Ci-Ci2)alkyl and a substituted (Ci-Ci2)alkyl;T2, T3, T4, T5and T6are each independently selected from (Ci-Ci2)alkyl, substituted (Ci-Ci2)alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA)W, (PEG)n, (AA)P, -(CR13OH)X-, 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, an acetal group, a disulfide, a hydrazine, and an ester; andV1, V2, V3, V4,V5and V6are each independently selected from a covalent bond, -CO-, -NR15-, -NR15(CH2)q-, -NR15(C6H4)-, -CONR15-, -NR15CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR15-, -NR15SO2- and -P(O)OH-, wherein q is an integer from 1 to 6;90NAI-5011166969vlwherein:(PEG)n is , where n is an integer from 1 to 30;EDA is an ethylene diamine moiety having the following structure:N4-amino-piperidine (4AP) is R12•AA is an amino acid residue, where p is an integer from 1 to 20; andeach R12is independently selected from hydrogen, an alkyl, a substituted alkyl, a polyethylene glycol moiety, an aryl and a substituted aryl, wherein any two adjacent R12groups may be cyclically linked to form a piperazinyl ring;each R13is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl; andeach R15is 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.

[0349] In some embodiments, LAcomprises:-(T1-V1)a-(T2-V2)b-(T3-V3)c-(T4-V4)d-(T5-V5)e-(T6-V6)f-, whereina, b, c, d, e and f are each independently 0 or 1, provided at least one of a, b, c, d, e, and f is 1;T1, T2, T3, T4, T5and T6are each independently selected from a covalent bond, (Ci-Cn)alkyl, substituted (Ci-Ci2)alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA)W, (PEG)n, (AA)P, -(CR13OH)X-, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxy-phenyl (PHP), an acetal group, a hydrazine, a disulfide, and an ester, wherein EDA is an ethylene diamine moiety, PEG is a polyethylene glycol, and AA is an amino acid residue or an amino acid analog, wherein each w is an91NAI-5011166969vlinteger 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;V1, V2, V3, V4,V5and V6are each independently selected from the group consisting of a covalent bond, -CO-, -NR15-, -NR15(CH2)q-, -NR15(C6H4)-, -CONR15-, -NR15CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR15-, -NR15SO2- and -P(O)OH-, wherein each q is an integer from 1 to 6;each R13is independently selected from hydrogen, an alkyl, a substituted alkyl, an aryl, and a substituted aryl; andeach R15is 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.

[0350] In some embodiments of LA:T1is selected from a (Ci-Ci2)alkyl and a substituted (Ci-Ci2)alkyl;T2, T3, T4, T5and T6are each independently selected from a covalent bond, (Ci-Cn)alkyl, substituted (Ci-Ci2)alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA)W, (PEG)n, (AA)p, -(CR13OH)X-, 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PAB A, PAP, PHP, an acetal group, a hydrazine, and an ester; andV1, V2, V3, V4,V5and V6are each independently selected from the group consisting of a covalent bond, -CO-, -NR15-, -NR15(CH2)q-, -NR15(C6H4)-, -CONR15-, -NR15CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2- , -SO2NR15-, -NR15SO2-, and -P(O)OH-; wherein:(PEG)nis , where n is an integer from 1 to 30;EDA is an ethylene diamine moiety having the following structure:N \R12, where y is an integer from 1 to 6 and r is 0 or 1;4-amino-piperidine (4AP) is92NAI-5011166969vleach R12is independently selected from hydrogen, an alkyl, a substituted alkyl, a polyethylene glycol moiety, an aryl and a substituted aryl, wherein any two adjacent R12groups may be cyclically linked to form a piperazinyl ring;a, b, c, and d are each 1; ande and f are 0.

[0351] In some embodiments, T1, T2, T3, T4, T5and T6are each optionally substituted with a glycoside.

[0352] In some embodiments, MABO, MABC, PABO, PABC, PAB, PABA, PAP and PHP are each optionally substituted with a glycoside.

[0353] In some embodiments, the glycoside is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.

[0354] In certain embodiments, T1, T2, T3, T4, T5and T6and V1, V2, V3, V4,V5and V6are selected from the following:wherein:T1is (Ci-Ci2)alkyl and V1is -CO-;T2is (AA)Pand V2is absent;T3is PABC and V3is absent;p is an integer from 1 to 10; andd, e and f are each 0; orwherein:T1is (Ci-Ci2)alkyl and V1is -CONH-;T2is (PEG)nand V2is -CO-;T3is (AA)Pand V3is absent;T4is PABC and V4is absent;p is an integer from 1 to 10; ande and f are each 0; orwherein:T1is (Ci-Ci2)alkyl and V1is -CO-;T2is an amino acid analog and V2is -NH-;T3is (PEG)nand V3is -CO-;T4is (AA)Pand V4is absent;T5is PABC and V5is absent;p is an integer from 1 to 10; andf is 0; or93NAI-5011166969vlwherein:T1is (Ci-Ci2)alkyl and V1is -CONH-;T2is (PEG)nand V2is -CO-;T3is (AA)Pand V3is absent;T4is PABC and V4is absent;p is an integer from 1 to 10; ande and f are each 0; orwherein:T1is (Ci-Ci2)alkyl and V1is -CONH-;T2is substituted (Ci-Ci2)alkyl and V2is -CO-; T3is (AA)Pand V3is absent;T4is PABC and V4is absent;p is an integer from 1 to 10; ande and f are each 0; orwherein:T1is (Ci-Ci2)alkyl and V1is -CONH-;T2is (PEG)nand V2is -CO-;T3is (AA)Pand V3is absent;T4is PABA and V4is -CO-;T5is (Ci-Ci2)alkyl and V5is absent;p is an integer from 1 to 10; andf is 0; orwherein:T1is (Ci-Ci2)alkyl and V1is -CO-;T2is 4AP and V2is -CO-;T3is (Ci-Ci2)alkyl and V3is -CO-;T4is (AA)Pand V4is absent;T5is PABC and V5is absent;p is an integer from 1 to 10; andf is 0; orwherein:T1is (Ci-Ci2)alkyl and V1is -CO-;T2is 4AP and V2is -CO-;T3is (Ci-Ci2)alkyl and V3is -O-;94 NAI-5011166969vlT4is (Ci-Ci2)alkyl and V4is -CO-;T5is (AA)Pand V5is absent;p is an integer from 1 to 10; andT6is PABC and V6is absent; orwherein:T1is (Ci-Ci2)alkyl and V1is -CO-;T2is an amino acid analog and V2is absent; T3is (AA)Pand V3is absent;T4is PABC and V4is absent;p is an integer from 1 to 10; ande and f are each 0; orwherein:T1is (Ci-Ci2)alkyl and V1is -CONH-;T2is (PEG)n and V2is -CONH-;T3is substituted (Ci-Ci2)alkyl and V3is -CO-; T4is (AA)Pand V4is absent;T5is PABC and V5is absent;p is an integer from 1 to 10; andf is 0; orwherein:T1is (Ci-Ci2)alkyl and V1is -CO-;T2is an (AA)Pand V2is -NH-;T3is (PEG)nand V3is -CO-;T4is (AA)Pand V4is absent;T5is PABC and V5is absent;p is an integer from 1 to 10; andf is 0; orwherein:T1is (Ci-Ci2)alkyl and V1is -CONH-;T2is (PEG)nand V2is -CO-;T3is (AA)Pand V3is absent;T4is PAP and V4is -C(O)O-;p is an integer from 1 to 10; ande and f are each 0; or95 NAI-5011166969vlwherein:T1is (Ci-Ci2)alkyl and V1is -CONH-;T2is substituted (Ci-Ci2)alkyl and V2is -CO-;T3is (AA)Pand V3is absent;T4is PABC and V4is absent;p is an integer from 1 to 10; ande and f are each 0; orwherein:T1is (Ci-Ci2)alkyl and V1is -CONH-;T2is substituted (Ci-Ci2)alkyl and V2is -CO-;T3is PABC and V3is absent; andd, e and f are each 0.

[0355] In certain embodiments, the left-hand side of the above linker structure for the first linker LAis attached to the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety, and the right-hand side of the above linker structure for the first linker LAis attached to the first drug or active agent.

[0356] As described above, in some embodiments, LBis a second linker comprising -(T7-V7)g-(T8-V8)h-(T9-V9)i-(T10-V10)j-(T11-V11)k-(T12-V12)i-(T13-V13)m-, where 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.

[0357] In some embodiments, in the second linker LB:T7is selected from a (Ci-Ci2)alkyl and a substituted (Ci-Ci2)alkyl;T8, T9, T10, T11, T12and T13are each independently selected from (Ci-Ci2)alkyl, substituted (Ci-Ci2)alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA)W, (PEG)n, (AA)P, -(CR13OH)X-, 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, an acetal group, a disulfide, a hydrazine, and an ester; andV7, V8, V9, V10,Vn, V12and V13are each independently selected from a covalent bond, -CO-, -NR15-, -NR15(CH2)q-, -NR15(C6H4)-, -CONR15-, -NR15CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR15-, -NR15SO2- and -P(O)OH-, wherein q is an integer from 1 to 6;wherein:(PEG)n is , where n is an integer from 1 to 30;96NAI-5011166969vlEDA is an ethylene diamine moiety having the following structure:N4-amino-piperidine (4AP) is R12•AA is an amino acid residue, where p is an integer from 1 to 20; andeach R12is independently selected from hydrogen, an alkyl, a substituted alkyl, a polyethylene glycol moiety, an aryl and a substituted aryl, wherein any two adjacent R12groups may be cyclically linked to form a piperazinyl ring;each R13is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl; andeach R15is 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.

[0358] Any convenient tether groups may be utilized for T7, T8, T9, T10, T11, T12and T13. For example, any of the tether groups described above in relation to T1, T2, T3, T4, T5and T6may be used for the tether groups T7, T8, T9, T10, T11, T12and T13.

[0359] Any convenient linking functional groups may be utilized for V7, V8, V9, V10,Vn, V12and V13. For example, any of the linking functional groups described above in relation to V1, V2, V3, V4, V5and V6may be used for the linking functional groups V7, V8, V9, V10,Vn, V12and V13.

[0360] In certain embodiments, each R13is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl. In these embodiments, alkyl, substituted alkyl, aryl, and substituted aryl are as described above for R13.

[0361] In certain embodiments, each R15is 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,97NAI-5011166969vlsubstituted cycloalkyl, heterocyclyl, and substituted heterocyclyl are as described above for R15. In these embodiments, various possible substituents are as described above for R15.

[0362] In certain embodiments of the second linker LB, one or more of the tether groups T7, T8, T9, T10, T11, T12and T13is each optionally substituted with a glycoside or glycoside derivative. In certain embodiments, the glycoside or glycoside derivative is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.

[0363] In certain embodiments of the second linker LB, the MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP tether structures shown above may be substituted with one or more additional groups selected from a glycoside and a glycoside derivative. For example, in some embodiments of the MABO, MABC, PABO, PABC, PAB, PABA, PAP, and PHP tether structures shown above, the phenyl ring may be substituted with one or more additional groups selected from a glycoside and a glycoside derivative. In certain embodiments, the glycoside or glycoside derivative is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.

[0364] In some embodiments, T7, T8, T9, T10, T11, T12and T13are each optionally substituted with a glycoside.

[0365] In some embodiments, MABO, MABC, PABO, PABC, PAB, PABA, PAP and PHP are each optionally substituted with a glycoside.

[0366] In some embodiments, the glycoside is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.

[0367] In some embodiments of LB:g, h, i, j, and k are each 1;1 and m is 0;T7is a covalent bond;T8, T9, T10, T11and T12are each independently selected from a covalent bond, (Ci-Cn)alkyl, substituted (Ci-Ci2)alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA)W, (PEG)n, (AA)p, -(CR13OH)X-, 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, an acetal group, a hydrazine, and an ester; andV7, V8, V9, V10,Vnand V12are each independently selected from the group consisting of a covalent bond, -CO-, -NR15-, -NR15(CH2)q-, -NR15(CeH4)-, -CONR15-, -NR15CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2- , -SO2NR15-, -NR15SO2-, and -P(O)OH-;wherein:98NAI-5011166969vl(PEG)nis , where n is an integer from 1 to 30;EDA is an ethylene diamine moiety having the following structure:NdR12, where y is an integer from 1 to 6 and r is 0 or 1;ND124-amino-piperidine (4AP) is R ; andeach R12is independently selected from hydrogen, an alkyl, a substituted alkyl, a polyethylene glycol moiety, an aryl and a substituted aryl, wherein any two adjacent R12groups may be cyclically linked to form a piperazinyl ring.

[0368] In some embodiments, T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, and T12are each optionally substituted with a glycoside.

[0369] In some embodiments, MABO, MABC, PABO, PABC, PAB, PABA, PAP and PHP are each optionally substituted with a glycoside.

[0370] In some embodiments, the glycoside is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.

[0371] In certain embodiments, T7, T8, T9, T10, T11, T12and T13and V7, V8, V9, V10,Vn, V12and V13are selected from the following:wherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CO-;T9is (AA)Pand V9is absent;T10is PABC and V10is absent; andk, 1 and m are each 0; orwherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CONH-;T9is (PEG)nand V9is -CO-;T10is (AA)Pand V10is absent; andT11is PABC and V11is absent; and1 and m are each 0; orwherein:99NAI-5011166969vlT7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CO-;T9is an amino acid analog and V9is -NH-;T10is (PEG)nand V10is -CO-;T11is (AA)Pand V11is absent;T12is PABC and V12is absent; andm is 0; orwherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CONH-;T9is (PEG)nand V9is -CO-;T10is (AA)Pand V10is absent;T11is PABC and V11is absent; and1 and m are each 0; orwherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CONH-;T9is substituted (Ci-Ci2)alkyl and V9is -CO-; T10is (AA)Pand V10is absent;T11is PABC and V11is absent; and1 and m are each 0; orwherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CONH-;T9is (PEG)nand V9is -CO-;T10is (AA)Pand V10is absent;T11is PABA and V11is -CO-;T12is (Ci-Ci2)alkyl and V12is absent; and m is 0; orwherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CO-;T9is 4AP and V9is -CO-;T10is (Ci-Ci2)alkyl and V10is -CO-;100 NAI-5011166969vlT11is (AA)Pand V11is absent;T12is PABC and V12is absent; andm is 0; orwherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CO-;T9is 4AP and V9is -CO-;T10is (Ci-Ci2)alkyl and V10is -O-;T11is (Ci-Ci2)alkyl and V11is -CO-;T12is (AA)Pand V12is absent; andT13PABC and V13is absent; orwherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CO-;T9is an amino acid analog and V9is absent;T10is (AA)Pand V10is absent;T11is PABC and V11is absent; and1 and m are each 0; orwherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CONH-;T9is (PEG)n and V9is -CONH-;T10is substituted (Ci-Ci2)alkyl and V10is -CO-; T11is (AA)Pand V11is absent;T12is PABC and V12is absent; andm is 0; orwherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CO-;T9is (AA)Pand V9is -NH-;T10is (PEG)nand V10is -CO-;T11is (AA)Pand V11is absent;T12is PABC and V12is absent; andm is 0; or101NAI-5011166969vlwherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CONH-;T9is (PEG)nand V9is -CO-;T10is (AA)Pand V10is absent;T11is PAP and V11is -C(O)O-; and1 and m are each 0; orwherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CO-;T9is (AA)Pand V9is absent;T10is PABC and V10is absent;T11is PAP and V11is -C(O)O-; and1 and m are each 0; orwherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CONH-;T9is substituted (Ci-Ci2)alkyl and V9is -CO-; T10is PABC and V10is absent; andk, 1 and m are each 0; orwherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is absent;T9is heteroaryl and V9is absent;T10is (Ci-Ci2)alkyl and V10is -CONH-;T11is (PEG)nand V11is -CO-; and1 and m are each 0; orwherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is absent;T9is heteroaryl and V9is absent;T10is (Ci-Ci2)alkyl and V10is -CONH-;T11is substituted (Ci-Ci2)alkyl and V11is -CO-; T12is (AA)Pand V12is absent; and102NAI-5011166969vlT13PAB and V13is absent; orwherein:T7is absent and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is absent;T9is heteroaryl and V9is absent;T10is (Ci-Ci2)alkyl and V10is -C0NH-;T11is substituted (Ci-Ci2)alkyl and V11is -CO-;T12is (AA)Pand V12is absent; andT13PABC and V13is absent.

[0372] In certain embodiments, the left-hand side of the above linker structure for the second linker LBis attached to the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety, and the right-hand side of the above linker structure for the second linker LBis attached to the second drug or active agent.

[0373] In certain embodiments, the conjugate is an antibody-drug conjugate where the TF antibody and the drugs are linked together by linkers as described above. In some instances, the linker (e.g., LAand / or LB) is a cleavable linker. A cleavable linker is a linker that includes one or more cleavable moieties, where the cleavable moiety includes one or more bonds that can dissociate under certain conditions, thus separating the cleavable linker into two or more separable portions. For example, the cleavable moiety may include one or more covalent bonds, which under certain conditions, can dissociate or break apart to separate the cleavable linker into two or more portions. As such the linkers that are included in an antibody-drug conjugate can be cleavable linkers, such that under appropriate conditions, the cleavable linker is cleaved to separate or release the drug from the antibody at a desired target site of action for the drug.

[0374] In some instances, a cleavable linker includes two cleavable moieties, such as a first cleavable moiety and a second cleavable moiety. The cleavable moieties can be configured such that cleavage of both cleavable moieties is needed in order to separate or release the drug from the TF antibody at a desired target site of action for the drug. For example, cleavage of a cleavable linker can be achieved by initially cleaving one of the two cleavable moieties and then cleaving the other of the two cleavable moieties. In certain embodiments, a cleavable linker includes a first cleavable moiety and a second cleavable moiety that hinders cleavage of the first cleavable moiety. By “hinders cleavage” is meant that the presence of an uncleaved second cleavable moiety reduces the likelihood or substantially inhibits the cleavage of the first cleavable moiety, thus substantially reducing the amount or preventing 103NAI-5011166969vlthe cleavage of the cleavable linker. For instance, the presence of uncleaved second cleavable moiety can hinder cleavage of the first cleavable moiety. The hinderance of cleavage of the first cleavable moiety by the presence of the second cleavable moiety, in turn, substantially reduces the amount or prevents the release of the drug from the antibody. For example, the premature release of the drug from the antibody can be substantially reduced or prevented until the antibody-drug conjugate is at or near the desired target site of action for the drug.

[0375] In some cases, since the second cleavable moiety hinders cleavage of the first cleavable moiety, cleavage of the cleavable linker can be achieved by initially cleaving the second cleavable moiety and then cleaving the first cleavable moiety. Cleavage of the second cleavable moiety can reduce or eliminate the hinderance on the cleavage of the first cleavable moiety, thus allowing the first cleavable moiety to be cleaved. Cleavage of the first cleavable moiety can result in the cleavable linker dissociating or separating into two or more portions as described above to release the drug from the antibody-drug conjugate. In some instances, cleavage of the first cleavable moiety does not substantially occur in the presence of an uncleaved second cleavable moiety. By substantially is meant that about 10% or less cleavage of the first cleavable moiety occurs in the presence of an uncleaved second cleavable moiety, such as 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 cleavage of the first cleavable moiety occurs in the presence of an uncleaved second cleavable moiety.

[0376] Stated another way, the second cleavable moiety can protect the first cleavable moiety from cleavage. For instance, the presence of uncleaved second cleavable moiety can protect the first cleavable moiety from cleavage, and thus substantially reduce or prevent premature release of the drug from the antibody until the antibody-drug conjugate is at or near the desired target site of action for the drug. As such, cleavage of the second cleavable moiety exposes the first cleavable moiety (e.g., deprotects the first cleavable moiety), thus allowing the first cleavable moiety to be cleaved, which results in cleavage of the cleavable linker, which, in turn, separates or releases the drug from the antibody at a desired target site of action for the drug as described above. In certain instances, cleavage of the second cleavable moiety exposes the first cleavable moiety to subsequent cleavage, but cleavage of the second cleavable moiety does not in and of itself result in cleavage of the cleavable linker ( / .< ., cleavage of the first cleavable moiety is still needed in order to cleave the cleavable linker).104NAI-5011166969vl

[0377] The cleavable moieties included in the cleavable linker may each be an enzymatically cleavable moiety. For example, the first cleavable moiety can be a first enzymatically cleavable moiety and the second cleavable moiety can be a second enzymatically cleavable moiety. An enzymatically cleavable moiety is a cleavable moiety that can be separated into two or more portions as described above through the enzymatic action of an enzyme. The enzymatically cleavable moiety can be any cleavable moiety that can be cleaved through the enzymatic action of an enzyme, such as, but not limited to, an ester, a peptide, a glycoside, and the like. In some instances, the enzyme that cleaves the enzymatically cleavable moiety is present at a desired target site of action, such as the desired target site of action of the drug that is to be released from the antibody-drug conjugate. In some cases, the enzyme that cleaves the enzymatically cleavable moiety is not present in a significant amount in other areas, such as in whole blood, plasma or serum. As such, the cleavage of an enzymatically cleavable moiety can be controlled such that substantial cleavage occurs at the desired site of action, whereas cleavage does not significantly occur in other areas or before the antibody-drug conjugate reaches the desired site of action.

[0378] For example, as described herein, antibody-drug conjugates of the present disclosure can be used for the treatment of cancer, such as for the delivery of a cancer therapeutic drug to a desired site of action where the cancer cells are present. In some cases, enzymes, such as an esterase that cleaves ester bonds or a glycosidase that cleaves glycosidic bonds, can be a biomarker for cancer that is overexpressed in cancer cells. The overexpression, and thus localization, of certain enzymes in cancer can be used in the context of the enzymatically cleavable moieties included in the cleavable linkers of the antibody-drug conjugates of the present disclosure to specifically release the drug at the desired site of action ( / .< ., the site of the cancer (and overexpressed enzyme)). Thus, in some embodiments, the enzymatically cleavable moiety is a cleavable moiety (e.g., an ester or a glycoside) that can be cleaved by an enzyme that is overexpressed in cancer cells. For instance, the enzyme can be an esterase. As such, in some instances, the enzymatically cleavable moiety is a cleavable moiety (e.g., an ester) that can be cleaved by an esterase enzyme. In some instances, the enzyme can be a glycosidase. As such, in some instances, the enzymatically cleavable moiety is a cleavable moiety (e.g., a glycoside or glycoside derivative) that can be cleaved by a glycosidase enzyme.

[0379] In certain embodiments, the enzymatically cleavable moiety is an ester bond. For example, the first cleavable moiety described above (i.e., the cleavable moiety protected from premature cleavage by the second cleavable moiety) can include an ester. The presence of 105NAI-5011166969vluncleaved second cleavable moiety can protect the first cleavable moiety (ester) from cleavage by an esterase enzyme, and thus substantially reduce or prevent premature release of the drug from the antibody until the antibody-drug conjugate is at or near the desired target site of action for the drug. In some instances, a portion of the linker adjacent to the first cleavable moiety is linked to or includes a substituent, where the substituent comprises the second cleavable moiety. In some instances, the second cleavable moiety includes a glycoside or glycoside derivative.

[0380] In some embodiments, the enzymatically cleavable moiety is sugar moiety, such as a glycoside (or glyosyl) or glycoside derivative. In some cases, the glycoside or glycoside derivative can facilitate an increase in the hydrophilicity of the cleavable linker as compared to a cleavable linker that does not include the glycoside or glycoside derivative. The glycoside or glycoside derivative can be any glycoside or glycoside derivative suitable for use in the cleavable linker and that can be cleaved through the enzymatic action of an enzyme. For example, the second cleavable moiety ( / .< ., the cleavable moiety that protects the first cleavable moiety from premature cleavage) can be a glycoside or glycoside derivative. For instance, in some embodiments, the first cleavable moiety includes an ester and the second cleavable moiety includes a glycoside or glycoside derivative. In certain embodiments, the second cleavable moiety is a glycoside or glycoside derivative selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc. In some instances, the second cleavable moiety is a glucuronide. In some instances, the second cleavable moiety is a galactoside. In some instances, the second cleavable moiety is a glucoside. In some instances, the second cleavable moiety is a mannoside. In some instances, the second cleavable moiety is a fucoside. In some instances, the second cleavable moiety is O-GlcNAc. In some instances, the second cleavable moiety is O-GalNAc.

[0381] The glycoside or glycoside derivative can be attached (covalently bonded) to the cleavable linker through a glycosidic bond. The glycosidic bond can link the glycoside or glycoside derivative to the cleavable linker through various types of bonds, such as, but not limited to, an O-glycosidic bond (an O-glycoside), an N-glycosidic bond (a glycosylamine), an S-glycosidic bond (a thioglycoside), or C-glycosidic bond (a C-glycoside or C-glycosyl). In some instances, the glycosidic bond is an O-glycosidic bond (an O-glycoside). In some cases, the glycoside or glycoside derivative can be cleaved from the cleavable linker it is attached to by an enzyme (e.g., through enzymatically-mediated hydrolysis of the glycosidic bond). A glycoside or glycoside derivative can be removed or cleaved from the cleavable linker by any convenient enzyme that is able to carry out the cleavage (hydrolysis) of the 106NAI-5011166969vlglycosidic bond that attaches the glycoside or glycoside derivative to the cleavable linker. An example of an enzyme that can be used to mediate the cleavage (hydrolysis) of the glycosidic bond that attaches the glycoside or glycoside derivative to the cleavable linker is a glycosidase, such as a glucuronidase, a galactosidase, a glucosidase, a mannosidase, a fucosidase, and the like. Other suitable enzymes may also be used to mediate the cleavage (hydrolysis) of the glycosidic bond that attaches the glycoside or glycoside derivative to the cleavable linker. In some cases, the enzyme used to mediate the cleavage (hydrolysis) of the glycosidic bond that attaches the glycoside or glycoside derivative to the cleavable linker is found at or near the desired site of action for the drug of the antibody-drug conjugate. For instance, the enzyme can be a lysosomal enzyme, such as a lysosomal glycosidase, found in cells at or near the desired site of action for the drug of the antibody-drug conjugate. In some cases, the enzyme is an enzyme found at or near the target site where the enzyme that mediates cleavage of the first cleavable moiety is found.

[0382] In some embodiments, a TF-ADC is represented by Formula (I):wherein:Ab represents the antibody that binds to TF;Z1, Z2, and Z4are each independently CR4;Z3is C-LB-W2;R1, R2, R3and R4are each selected from hydrogen and (Ci-Ci2)alkyl;LAis a first linker wherein:T1is (Ci-Ci2)alkyl and V1is -C0NH-;T2is substituted (Ci-Ci2)alkyl and V2is -CO-;T3is (AA)Pwhere p is an integer from 1-20 and V3is a covalent bond;T4is PABC and V4is a covalent bond;a, b, c, and d are each 1;e and f are each 0; andLBis a second linker wherein107NAI-5011166969vlT7is a covalent bond and V7is -NHCO-;T8is (Ci-Ci2)alkyl and V8is -CONH-;T9is substituted (Ci-Ci2)alkyl and V9is -CO-;T10is (AA)Pwhere p is an integer from 1-20 and V10is a covalent bond;T11is PABC and V11is a covalent bond; andh, i, j, and k are each 1; and1 and m are each 0;s is an integer from 1 to 10;W1is a first drug; andW2is a second drug.

[0383] In some embodiments, one or both of W1and W2are camptothecin analogues, for example, belotecan.

[0384] In some embodiments, a TF-ADC is represented by Formula (I):wherein:Ab represents the antibody that binds to TF;Z1, Z2, and Z4are each independently CR4;Z3is C-LB-W2;R1, R2, R3and R4are each selected from hydrogen and (Ci-Ci2)alkyl;LAis a linker wherein:T1is (Ci-Ce)alkyl and V1is -CONH-;T2is (Ci-Ce)alkylene substituted with -NHCO(PEG)k’, wherein k’ is an integer from 2 to 10 and V2is -CO-;T3is (AA)2 and V3is a covalent bond;T4is PABC substituted with a glycoside and V4is a covalent bond;a, b, c, and d are each 1; ande and f are each 0; andLBis a linker wherein108NAI-5011166969vlT7is a covalent bond and V7is -NHCO-;T8is (Ci-Ce)alkyl and V8is -CONH-;T9is (Ci-Ce)alkylene substituted with -NHCO(PEG)k’, wherein k’ is an integer from 2 to 10 and V9is -CO-;T10is (AA)2 and V10is a covalent bond;T11is PABC substituted with a glycoside and V11is a covalent bond;h, i, j, and k are each 1; and1 and m are each 0;s is an integer from 1 to 10;W1is a first drug; andW2is a second drug.

[0385] In some embodiments, s is 2. In some embodiments, s is 4.

[0386] In some embodiments, the PABC of one or both of T4and T11is substituted with a glucuronide. In some embodiments, one or both of T1and T8is ethyl. In some embodiments, one or both of T2and T9is Cs alkylene substituted with -NHCO(PEG)k’, where k’ is an integer from 5 to 10. In some embodiments, one or both of W1and W2are camptothecin analogues, for example, belotecan.

[0387] In some embodiments, a TF-ADC as disclosed herein comprises any payload, any linker, or any linker-payload as disclosed in US Patent Application No. 2022-0241423 and International Publication No. WO 2022187370, each of which is incorporated herein by reference in its entirety.

[0388] In some embodiments, the TF-ADC is represented by Formula (II):109NAI-5011166969vlAbs(II), wherein:Ab represents the antibody that binds to TF; ands is an integer from 1 to 10.

[0389] In some embodiments, s is an integer from 1 to 4. In some embodiments, s is 2. In some embodiments, s is 4.

[0390] In some embodiments, a TF-ADC is produced by conjugating a linker-payload of Formula (Ila), shown below, with a TF antibody (Ab). In further embodiments, a TF-ADC is prepared by conjugating two linker-payloads of Formula (Ila) with one TF antibody (Ab). Accordingly, the TF-ADC has a drug-to-antibody ratio (DAR) of 4. In some embodiments, a TF-ADC is prepared by conjugating four linker-payloads of Formula (Ila) with one TF antibody (Ab). Accordingly, the TF-ADC has a drug-to-antibody ratio (DAR) of 8.110NAI-5011166969vl

[0391] Any of the chemical entities, linkers and conjugation moieties set forth in the structures above may be adapted for use in the subject compounds and conjugates.

[0392] Additional disclosure related to hydrazinyl-indolyl and hydrazinyl-pyrrolo-pyridinyl compounds and methods for producing a conjugate is found in U.S. Patent No. 9,310,374 and U.S. Patent No. 9,493,413, the disclosures of each of which are incorporated herein by reference.TISSUE FACTOR (TF) ANTIBODIES

[0393] As noted above, a subject conjugate comprises an antibody (Ab) that binds to TF. The amino acid sequence of the antibody can be modified to include a 2-formylglycine (fGly) residue. 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, such as: 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 He 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 Gin or Q; Arginine or Arg or R; Serine or Ser or S;Threonine or Thr or T; Valine or Vai or V; Tryptophan or Trp or W; and Tyrosine or Tyr or Y.illNAI-5011166969vl

[0394] TF-ADCs described herein comprise a drug and TF antibody conjugated thereto. In some embodiments, a TF antibody refers to an antibody, specifically binding to TF, such as 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 human constant regions) that binds to TF. In some embodiments, a TF antibody can bind to TF which is expressed on the surface of a mammalian (e.g., human) cell, including a TF-expressing tumor cell. In some embodiments, a TF antibody binds a TF extracellular epitope expressed on a cell such as a tumor cell (e.g., an extracellular TF epitope). In some embodiments, TF is a human TF. An exemplary amino acid sequence of human TF is described herein (SEQ ID NO: 175).

[0395] In some embodiments, the TF antibody competes for binding to TF with a reference TF antibody that comprises a 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, such as an amino acid sequence of a VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 as described in any one of Tables 1-2. Accordingly, in some embodiments, the 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 as EXMA-006; or (b) the antibody designated as EXMA-007. In some embodiments, a TF-ADC comprises a drug conjugated (directly or indirectly) to a TF antibody that competes for binding to TF with a reference TF antibody that comprises one, two, and / or three VH CDRs and one, two, and / or three VL CDRs from: (a) the antibody designated as EXMA-006; or (b) the antibody designated as EXMA-007. In some embodiments, the TF antibody competes for binding to TF with a reference TF antibody that comprises a VH region and VL region from: (a) the antibody designated as EXMA-006; or (b) the antibody designated as EXMA-007. In some embodiments, the TF antibody competes for binding to TF with a reference TF antibody that comprises: (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 can be used, see, for example, International Publication Nos. WO2019136309 and W02021003399, each of which in incorporated herein by reference in its entirety.

[0396] In some embodiments, the TF antibody comprises a VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 of any one of the 112NAI-5011166969vlantibodies described herein, such as an amino acid sequence of a VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 as described in any one of Tables 1-2. Accordingly, 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 as EXMA-006; or (b) the antibody designated as 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 as EXMA-006; or (b) the antibody designated as EXMA-007.

[0397] In some embodiments, the TF antibody comprises a VH region, which comprises one or more (such as one, two, or three) of VH CDR1, VH CDR2, and VH CDR3 as described herein, such as in any one of Tables 1-2; and / or a VL region, which comprises one or more (such as one, two, or three) of VL CDR1, VL CDR2, and VL CDR3 as described herein, such as in any one of Tables 1-2. In some embodiments, the TF antibody is bispecific and comprises a first binding region that comprises one, two, and / or three heavy chain CDRs and / or one, two, and / or three light chain CDRs as described in any one of Tables 1-2 and a second region that comprises 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 that comprises one, two, and / or three heavy chain CDRs and / or one, two, and / or three light chain CDRs as described in any one of Tables 1-2 and a second binding domain that comprises 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.

[0398] The antibody designated as EXMA-006 comprises a VH amino acid sequence of SEQ ID NO:25 and a VL amino acid sequence of SEQ ID NO:26.

[0399] The antibody designated as EXMA-007 comprises a VH amino acid sequence of SEQ ID NO:41 and a VL amino acid sequence of SEQ ID NO:42.113NAI-5011166969vlTable 1: Antibody EXMA-006-CDR SequencesExemplary* Kabat Chothia AbM Contact IMGT VH GYTFDVYGIS VYGIS GYTFDVY GYTFDVYGIS DVYGIS GYTFDVYG CDR1 (SEQ ID NO:1) (SEQ ID NO:7) (SEQ ID NO: 8) (SEQ ID NO:1) (SEQ ID NO: 15) (SEQ ID NO:21) WIAPYSGNTNYA WIAPYSGNTNYA WMGWIAPYSGN VH PYSG WIAPYSGNTN IAPYSGNT VH CDR QKLQG QKLQG TN CDR2 (SEQ ID NOV) (SEQ ID NO: 14) (SEQ ID NO:22) Seq. (SEQ ID NO:2) (SEQ ID NO:2) (SEQ ID NO: 16)DAGTYSPFGYGM DAGTYSPFGYGM DAGTYSPFGYGM ARDAGTYSPFGY ARDAGTYSPFGY VH AGTYSPFGYGMD DV DV DV GMD GMDV CDR3 (SEQ ID NO: 10)(SEQ ID NO:3) (SEQ ID NO:3) (SEQ ID NO:3) (SEQ ID NO: 17) (SEQ ID NO:23) VL QASQSINNWLA QASQSINNWLA SQSINNW QASQSINNWLA NNWLAWY QSINNW CDR1 (SEQ ID NO:4) (SEQ ID NO:4) (SEQ ID NO: 11) (SEQ ID NO:4) (SEQ ID NO: 18) (SEQ ID NO:24) VL CDR VL KAYNLES KAYNLES KAY KAYNLES LLIYKAYNLE KAYSeq. CDR2 (SEQ ID NO:5) (SEQ ID NO:5) (SEQ ID NO: 12) (SEQ ID NO:5) (SEQ ID NO: 19) (SEQ ID NO: 12)VL QLFQSLPPFT QLFQSLPPFT FQSLPPF QLFQSLPPFT QLFQSLPPF QLFQSLPPFT CDR3 (SEQ ID NO:6) (SEQ ID NO:6) (SEQ ID NO: 13) (SEQ ID NO:6) (SEQ ID NO:20) (SEQ ID NO:6) VH Sequence*:QVQLVQSGAEVKKPGASVKVSCKASGYTFDVYGISWVRQAPGQGLEWMGWIAPYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDT AVYYCARDAGTYSPFGYGMDVWGQGTTVTVSS (SEQ ID NO:25)VL Sequence*:DIQMTQSPSTLSASVGDRVTITCQASQSINNWLAWYQQKPGKAPKLLIYKAYNLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQLFQSLPPFTFGGGTKVEIK (SEQ ID NO:26)* Exemplary CDR sequences encompass amino acids as determined by Kabat plus Chothia114NAI-5011166969vlTable 2: Antibody EXMA-007-CDR SequencesExemplary* Kabat Chothia AbM Contact IMGT VH GYTFDAYGIS AYGIS GYTFDAY GYTFDAYGIS DAYGIS GYTFDAYG CDR1 (SEQ ID NO:27) (SEQ ID NO:31) (SEQ ID NO:32) (SEQ ID NO:27) (SEQ ID NO:35) (SEQ ID NO:39) WIAPYSGNTNYA WIAPYSGNTNYA WMGWIAPYSGN VH PYSG WIAPYSGNTN IAPYSGNT VH CDR QKLQG QKLQG TN CDR2 (SEQ ID NOV) (SEQ ID NO: 14) (SEQ ID NO:22) Seq. (SEQ ID NO: 2) (SEQ ID NO:2) (SEQ ID NO: 16)DAGTYSPFGYGM DAGTYSPFGYGM DAGTYSPFGYGM ARDAGTYSPFGY ARDAGTYSPFGY VH AGTYSPFGYGMD DV DV DV GMD GMDV CDR3 (SEQ ID NO: 10)(SEQ ID NO:3) (SEQ ID NO:3) (SEQ ID NO:3) (SEQ ID NO: 17) (SEQ ID NO:23) VL RASESISNWLA RASESISNWLA SESISNW RASESISNWLA SNWLAWY ESISNW CDR1 (SEQ ID NO:28) (SEQ ID NO:28) (SEQ ID NO:33) (SEQ ID NO:28) (SEQ ID NO:36) (SEQ ID NO:40) VL CDR VL KAYSLEY KAYSLEY KAY KAYSLEY LLIYKAYSLE KAYSeq. CDR2 (SEQ ID NO:29) (SEQ ID NO:29) (SEQ ID NO: 12) (SEQ ID NO:29) (SEQ ID NO:37) (SEQ ID NO: 12)VL QQFQKLPPFT QQFQKLPPFT FQKLPPF QQFQKLPPFT QQFQKLPPF QQFQKLPPFT CDR3 (SEQ ID NO:30) (SEQ ID NO:30) (SEQ ID NO:34) (SEQ ID NO:30) (SEQ ID NO:38) (SEQ ID NO:30) VH Sequence*:QVQLVQSGAEVKKPGASVKVSCKASGYTFDAYGISWVRQAPGQGLEWMGWIAPYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDT AVYYCARDAGTYSPFGYGMDVWGQGTTVTVSS (SEQ ID NO:41)VL Sequence*:DIQMTQSPSTLSASVGDRVTITCRASESISNWLAWYQQKPGKAPKLLIYKAYSLEYGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQFQKLPPFTFGGGTKVEIK (SEQ ID NO:42)* Exemplary CDR sequences encompass amino acids as determined by Kabat plus Chothia115NAI-5011166969vl

[0400] 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.

[0401] In some embodiments, the TF antibody comprises a heavy chain having a combination of (i) a VH as described herein, such as in any one of Tables 1-2, and (ii) one or more heavy chain constant regions (e.g., CHI, hinge, CH2, and CH3). An exemplary IgG heavy chain can comprise any VH amino acid sequence as described herein and the following CHI, hinge, CH2, and CH3 amino acid sequence:ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAK TKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR EPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO:89).

[0402] In some embodiments, the TF antibody comprises a heavy chain having a combination of (i) a VH as described herein, such as in any one of Tables 1-2, and (ii) one or more heavy chain constant regions (e.g., CHI, hinge, CH2, and CH3). An exemplary IgG heavy chain can comprise any VH amino acid sequence as described herein and the following CHI, hinge, CH2, and CH3 amino acid sequence:ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAK TKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR EPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NOVO).

[0403] In some embodiments, a TF antibody comprises a light chain having a combination of (i) a VL region as described herein, such as 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:RTVAAPSVFIFPPSDSQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:95).116NAI-5011166969vl

[0404] In some embodiments, a TF antibody comprises a light chain having a combination of (i) a VL region as described herein, such as 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:RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 96)

[0405] In some embodiments, the TF antibody comprises (a) a heavy chain having a combination of (i) a VH as described herein, such as in any one of Tables 1-2, and (ii) one or more heavy chain constant regions (e.g., CHI, hinge, CH2, and CH3); and (b) a light chain having a combination of (i) a VL as described herein, such as in any one of Tables 1-2, and (ii) a light chain constant region in an IgG format (CL1).

[0406] In some embodiments, the TF antibody comprises a VH having an amino acid sequence of:QVQLVQSGAEVKKPGASVKVSCKASGYTFDVYGISWVRQAPGQGLEWMGWIAPYS GNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDAGTYSPFGYGM DVWGQGTTVTVSS (SEQ ID NO:25); and a VL having an amino acid sequence of:DIQMTQSPSTLSASVGDRVTITCQASQSINNWLAWYQQKPGKAPKLLIYKAYNLESG VPSRFSGSGSGTEFTLTISSLQPDDFATYYCQLFQSLPPFTFGGGTKVEIK (SEQ ID NO:26).

[0407] In some embodiments, the TF antibody comprises a VH having an amino acid sequence of:QVQLVQSGAEVKKPGASVKVSCKASGYTFDAYGISWVRQAPGQGLEWMGWIAPYS GNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDAGTYSPFGYGM DVWGQGTTVTVSS (SEQ ID NO:41); and a VL having an amino acid sequence of:DIQMTQSPSTLSASVGDRVTITCRASESISNWLAWYQQKPGKAPKLLIYKAYSLEYG VPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQFQKLPPFTFGGGTKVEIK (SEQ ID NO:42)

[0408] In some embodiments, the antibody that binds to TF comprises a heavy chain having an amino acid sequence of:QVQLVQSGAEVKKPGASVKVSCKASGYTFDVYGISWVRQAPGQGLEWMGWIAPYS GNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDAGTYSPFGYGM DVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG117NAI-5011166969vlALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW YVDGVEVHNAI<TI<PREEQYNSTYRVVSVLTVLHQDWLNGI<EYI<CI<VSNI<ALPAPI EKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO:93); and a light chain having an amino acid sequence of:DIQMTQSPSTLSASVGDRVTITCQASQSINNWLAWYQQKPGKAPKLLIYKAYNLESG VPSRFSGSGSGTEFTLTISSLQPDDFATYYCQLFQSLPPFTFGGGTKVEIKRTVAAPSV FIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDST YSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:91).

[0409] In some embodiments, the antibody that binds to TF comprises a heavy chain having an amino acid sequence of:QVQLVQSGAEVKKPGASVKVSCKASGYTFDAYGISWVRQAPGQGLEWMGWIAPYS GNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDAGTYSPFGYGM DVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW YVDGVEVHNAI<TI<PREEQYNSTYRVVSVLTVLHQDWLNGI<EYI<CI<VSNI<ALPAPI EKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 94); and a light chain having an amino acid sequence of:DIQMTQSPSTLSASVGDRVTITCRASESISNWLAWYQQKPGKAPKLLIYKAYSLEYG VPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQFQKLPPFTFGGGTKVEIKRTVAAPSV FIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDST YSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:92).

[0410] In some embodiments, one or both of the heavy chains of the TF antibody further comprises a signal peptide, for example 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, for example at the N terminus of the chain. In some embodiments, the signal peptide comprises an amino acid sequence of MMSFVSLLLVGILFHATQA (SEQ ID NO:97). In some embodiments, the signal peptide comprises an amino acid sequence of MGWSLILLFLVAVATRVHS (SEQ ID NO:98).

[0411] In some embodiments, the TF antibody comprises a heavy chain with a signal peptide and therefore have an amino acid sequence of:118NAI-5011166969vlMMSFVSLLLVGILFHATOAOVQLVQSGAEVKKPGASVKVSCKASGYTFDVYGISWV RQAPGQGLEWMGWIAPYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTA VYYCARDAGTYSPFGYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAAL GCLVKDYFPEP VT VSWNSGALTSGVHTFP AVLQ SSGL YSLS S VVTVPS S SLGTQT YIC NVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPG (SEQ ID NO: 179), wherein the underlined amino acids represent the signal peptide sequence.

[0412] In some embodiments, the TF antibody comprises a heavy chain with a signal peptide and therefore have an amino acid sequence of:MMSFVSLLLVGILFHATOAOVOLVQSGAEVKKPGASVKVSCKASGYTFDAYGISW VRQAPGQGLEWMGWIAPYSGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDD TAVYYCARDAGTYSPFGYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCL VKDYFPEPVTVSWNSGALTSGVHTFP A VLQS SGL YSLS S VVTVPS S SLGTQ TYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLT CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNV FSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 180), wherein the underlined amino acids represent the signal peptide sequence.

[0413] In some embodiments, a TF antibody comprises one or more CDRs (e.g., one, two, three, four, five, or six CDRs), for example, a VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 as described in Table 1. In some embodiments, a TF antibody comprises one or more CDRs, (e.g, one, two, three, four, five, or six CDRs), for example, a VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 as described in Table 2

[0414] In some embodiments, a TF antibody comprises one or more CDRs, (e.g, one, two, or three VH CDRs), for example, a VH CDR1, VH CDR2, VH CDR3, as described in Table 1. In some embodiments, a TF antibody comprises one or more CDRs, (e.g., one, two, or three VL CDRs), for example, a VL CDR1, VL CDR2, and / or VL CDR3, as described in Table 1. In some embodiments, a TF antibody comprises one or more CDRs, (e.g., one, two, or three VH CDRs), for example, a VH CDR1, VH CDR2, VH CDR3, as described in 119NAI-5011166969vlTable 1 and one or more CDRs, (e.g., one, two, or three VL CDRs), for example, a VL CDR1, VL CDR2, and / or VL CDR3, as described in Table 1.

[0415] In some embodiments, a TF antibody comprises one or more CDRs, (e.g., one, two, or three VH CDRs), for example, a VH CDR1, VH CDR2, VH CDR3, as described in Table 2. In other embodiments, a TF antibody comprises one or more CDRs, (e.g., one, two, or three VL CDRs), for example, a VL CDR1, VL CDR2, and / or VL CDR3, as described in Table 2. In some embodiments, a TF antibody comprises one or more CDRs, (e.g., one, two, or three VH CDRs), for example, a VH CDR1, VH CDR2, VH CDR3, as described in Table 2 and one or more CDRs, (e.g., one, two, or three VL CDRs), for example, a VL CDR1, VL CDR2, and / or VL CDR3, as described in Table 2.

[0416] In some embodiments, a TF antibody comprises one or more (e.g., one, two, or three) VH CDRs as described herein, such as in Tables 1-2. In other embodiments, a TF antibody comprises one or more (e.g., one, two, or three) VL CDRs as described herein, such as in Tables 1-2. In some embodiments, a TF antibody comprises one or more (e.g., one, two, or three) VH CDRs as described herein, such as in Tables 1-2 and one or more (e.g., one, two, or three) VL CDRs as described herein, such as in Tables 1-2. Accordingly, in some embodiments, a TF antibody comprises a VH CDR1 comprising an amino acid sequence comprising any one of SEQ ID NOs:l, 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, a TF antibody comprises a VH CDR3 comprising an amino acid sequence of any one of SEQ ID NOs:3, 10, 17, and 23. In some embodiments, a TF antibody comprises a VH CDR1 and / or a VH CDR2 and / or a VH CDR3 independently selected from a VH CDR1, VH CDR2, VH CDR3 as described in any one of Tables 1-2. In some embodiments, a TF antibody comprises a VL CDR1 comprising an amino acid sequence of any one of SEQ ID NOs:4, 11, 18, 24, 28, 33, 36, and 40. In some embodiments, a TF antibody comprises a VL CDR2 comprising an amino acid sequence of any one of SEQ ID NOs:5, 12, 19, 29, and 37. In some embodiments, a TF antibody comprises a VL CDR3 comprising an amino acid sequence of any one of SEQ ID NOs:6, 13, 20, 30, 34, and 38. In some embodiments, a 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 as described herein, such as in any one of Tables 1-2.

[0417] 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) the antibody designated as EXMA-006 that comprises a VH amino acid sequence of SEQ ID 120NAI-5011166969vlNO:25 and a VL amino acid sequence of SEQ ID NO:26; or (b) the antibody designated as EXMA-007 that comprises a VH amino acid sequence of SEQ ID NO:41 and a VL amino acid sequence of SEQ ID NO:42. 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 as 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 as EXMA-006.

[0418] In some embodiments, a TF-ADC comprises a TF antibody, wherein the antibody comprises: (a) a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 amino acid sequence as described herein, such as in Tables 1-2; and / or (b) a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 amino acid sequence as described herein, such as in Tables 1-2. In some embodiments, a TF-ADC comprises a TF antibody wherein the antibody comprises: a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 amino acid sequence as described herein, such as in Tables 1-2. In some embodiments, a TF-ADC comprises a TF antibody, wherein the antibody comprises a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 amino acid sequence as described herein, such as in Tables 1-2

[0419] 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 IDNO:6.

[0420] 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 IDNO:6.

[0421] 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 NOTO, a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 11, a VL CDR2 comprising the121NAI-5011166969vlamino acid sequence of SEQ ID NO: 12, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 13.

[0422] 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 IDNO:6.

[0423] 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.

[0424] 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.

[0425] 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.

[0426] 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.

[0427] 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 122NAI-5011166969vlSEQ ID N0:9, 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.

[0428] 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.

[0429] 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.

[0430] 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.

[0431] In some embodiments, a TF-ADC comprises a TF antibody, wherein the antibody comprises a VH region and / or VL region described herein, wherein an VH and / or VL comprises human framework sequences. In some embodiments, an VH region and / or VL region comprises a framework 1 (FR1), a framework 2 (FR2), a framework 3 (FR3) and / or a framework 4 (FR4) sequence, such as a human FR1, a human FR2, a human FR3 and / or a human FR4.

[0432] In some embodiments, an CDRs of a TF antibody can be determined by 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, U.S. Department of Health and Human Services, NIH Publication No. 91-3242).

[0433] In some embodiments, CDRs of a TF antibody can be determined by the Chothia system, which will be referred to herein as the “Chothia CDRs” (see, e.g., Chothia and Lesk,123NAI-5011166969vl1987, 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).

[0434] In some embodiments, CDRs of a TF antibody can be determined by the ImMunoGeneTics (IMGT®) system, for example, 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 (“IMGT® CDRs”).

[0435] In some embodiments, CDRs of a TF antibody can be determined by the AbM system, which will be referred to herein as the “AbM CDRs,” for example 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 Diibel, eds., Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001).

[0436] In some embodiments, CDRs of a TF antibody can be determined by the Contact system, which will be referred to herein as the “Contact CDRs” (see, e.g., MacCallum RM et al., 1996, J Mol Biol 5:732-745). The Contact CDRs are based on an analysis of the available complex crystal structures.

[0437] In some embodiments, a TF antibody comprises a heavy chain variable region (VH) comprising: (1) a VH CDR1 comprising an 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 an amino acid sequence of any one of: (i) SEQ ID NO:2, (ii) SEQ ID NOV, (iii) SEQ ID NO: 14, (iv) SEQ ID NO: 16, and (v) SEQ ID NO:22; and (3) a VH CDR3 comprising an 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 comprising: (1) a VL CDR1 comprising an 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 an amino acid sequence of any one of: (i) SEQ ID NO:5, (ii) SEQ ID NO: 12, and (iii) SEQ ID NO: 19; and (3) a VL CDR3 comprising an amino acid sequence of any one of: (i) SEQ ID NO:6, (ii) SEQ ID NO: 13, and (iii) SEQ IDNO:20.

[0438] In some embodiments, a TF antibody comprises a VH region comprising: (1) a VH CDR1 comprising an amino acid sequence of any one of: (i) SEQ ID NO:1, (ii) SEQ ID NOV, (iii) SEQ ID NO:8, (iv) SEQ ID NO: 15, and (v) SEQ ID NO:21; (2) a VH CDR2 comprising an amino acid sequence of any one of: (i) SEQ ID NO:2, (ii) SEQ ID NOV, (iii) SEQ ID NO: 14, (iv) SEQ ID NO: 16, and (v) SEQ ID NO:22; and (3) a VH CDR3124NAI-5011166969vlcomprising an 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.

[0439] In some embodiments, a TF antibody comprises a VL region comprising: (1) a VL CDR1 comprising an 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 an amino acid sequence of any one of: (i) SEQ ID NO:5, (ii) SEQ ID NO: 12, and (iii) SEQ ID NO: 19; and (3) a VL CDR3 comprising an amino acid sequence of any one of: (i) SEQ ID NO:6, (ii) SEQ ID NO: 13, and (iii) SEQ ID NO:20.

[0440] In some embodiments, a TF-ADC comprises a TF antibody, wherein the antibody comprises: (1) a VL CDR1 comprising an amino acid sequence of any one of: SEQ ID NOs:4, 11, 18, and 24; (2) a VL CDR2 comprising an amino acid sequence of any one of: SEQ ID NOs:5, 12, and 19; and (3) a VL CDR3 comprising an amino acid sequence of any one of: SEQ ID NOs:6, 13, and 20.

[0441] In some embodiments, a TF-ADC comprises a TF antibody, wherein the antibody comprises a VL region comprising: (1) a VL CDR1 comprising an amino acid sequence of SEQ ID NO:4; (2) a VL CDR2 comprising an amino acid sequence of SEQ ID NO:5; and (3) a VL CDR3 comprising an amino acid sequence of SEQ ID NO:6.

[0442] In some embodiments, a TF-ADC comprises a TF antibody, wherein the antibody comprises: (a) a VH region comprising: (1) a VH CDR1 comprising an amino acid sequence of SEQ ID NO:1; and (b) a VL region comprising: (1) a VL CDR1 comprising an amino acid sequence of any one of SEQ ID NOs:4, 11, 18, and 24; (2) a VL CDR2 comprising an amino acid sequence of any one of SEQ ID NOs:5, 12, and 19; and (3) a VL CDR3 comprising an amino acid sequence of any one of SEQ ID NOs:6, 13, and 20.

[0443] In some embodiments, a TF-ADC comprises a TF antibody, wherein the antibody comprises: (a) a VH region comprising: (1) a VH CDR1 comprising an amino acid sequence of SEQ ID NO:1; and (b) a VL region comprising: (1) a VL CDR1 comprising an amino acid sequence of SEQ ID NO:4; (2) a VL CDR2 comprising an amino acid sequence of SEQ ID NO:5; and (3) a VL CDR3 comprising an amino acid sequence of SEQ ID NO:6.

[0444] In some embodiments, a TF-ADC is prepared from a TF antibody, wherein the antibody comprises a VH comprising an amino acid sequence of SEQ ID NO:25, and / or a VL comprising an amino acid sequence of SEQ ID NO:26.

[0445] In some embodiments, a TF-ADC is prepared from a TF antibody, wherein the antibody comprises a heavy chain that is inserted with one or more fGly site(s) and comprises an amino acid sequence of SEQ ID NO:78 or 81 and / or a light chain comprising an amino 125NAI-5011166969vlacid sequence of SEQ ID NO:26. In some embodiments, a TF-ADC is prepared from a TF antibody, wherein the antibody comprises a heavy chain that is inserted with one or more fGly site(s) and comprises an amino acid sequence of SEQ ID NO:78 or 81 and / or a light chain comprising an amino acid sequence of SEQ ID NO:91.

[0446] In some embodiments, a TF-ADC comprises a TF antibody, wherein the antibody comprises a heavy chain that is inserted with one or more fGly site(s) conjugated to a linkerdrug construct as disclosed herein and comprises an amino acid sequence of SEQ ID NO:84 or 87, and / or a light chain comprising an amino acid sequence of SEQ ID NO:26. In some embodime...

Claims

WHAT IS CLAIMED IS:

1. A method of treating a colorectal cancer (CRC) or a non-small cell lung cancer (NSCLC) in a subject, the method comprising administering an antibody-drug conjugate (ADC) of Formula (I), wherein the ADC comprises:a. an antibody that binds to tissue factor (TF); andb. two or more drugs conjugated to a pyridazine-pyrrolo coupling moiety, each via a linkerwherein:Ab represents the antibody that binds to TF;Z1, Z2, and Z4are each independently CR4;Z3is C-LB-W2;R1, R2, R3, and R4are each selected from hydrogen and alkyl;LAis a first linker comprising:-(T1-V1)a-(T2-V2)b-(T3-V3)c-(T4-V4)d-(T5-V5)e-(T6-V6)f-, wherein:a, b, c, d, e and f are each independently 0 or 1, provided at least one of a, b, c, d, e, and f are 1 ;T1, T2, T3, T4, T5and T6are each independently selected from a covalent bond, (Ci-Cn)alkyl, substituted (Ci-Ci2)alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA)W, (PEG)n, (AA)P, -(CR13OH)X-, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxy-phenyl (PHP), an acetal group, a hydrazine, a194NAI-5011166969vldisulfide, and an ester, wherein EDA is an ethylene diamine moiety, PEG is a polyethylene glycol, and AA is an amino acid residue or an amino acid analog, wherein 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;V1, V2, V3, V4,V5and V6are each independently selected from the group consisting of a covalent bond, -CO-, -NR15-, -NR15(CH2)q-, -NR15(C6H4)-, -CONR15-, -NR15CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR15-, -NR15SO2- and -P(O)OH-, wherein each q is an integer from 1 to 6;each R13is independently selected from hydrogen, an alkyl, a substituted alkyl, an aryl, and a substituted aryl; andeach R15is 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;LBis a second linker comprising:-(T7-V7)g-(T8-V8)h-(T9-V9)i-(T10-V10)j-(T11-V11)k-(T12-V12)i-(T13-V13)m-, wherein: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;T7, T8, T9, T10, T11, T12and T13are each independently selected from a covalent bond, (Ci-Ci2)alkyl, substituted (Ci-Ci2)alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA)w, (PEG)n, (AA)P, -(CR13OH)X-, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxy-phenyl (PHP), an acetal group, a hydrazine, a disulfide, and an ester, wherein EDA is an ethylene diamine moiety, PEG is a polyethylene glycol, and AA is an amino acid residue or an amino acid analog, wherein 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;V7, V8, V9, V10,Vn, V12and V13are each independently selected from the group consisting of a covalent bond, -CO-, -NR15-, -NR15(CH2)q-, -NR15(CeH4)-, -CONR15-, -195NAI-5011166969vlNR15CO-, -C(0)0-, -0C(0)-, -0-, -S-, -S(0)-, -SO2-, -SO2NR15-, -NR15SO2- and -P(O)OH-, wherein each q is an integer from 1 to 6;each R13is independently selected from hydrogen, an alkyl, a substituted alkyl, an aryl, and a substituted aryl; andeach R15is 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;W1is a first drug; andW2is a second drug.

2. The method of claim 1, wherein one or both of W1and W2is a camptothecin analog.

3. The method of claim 1 or 2, wherein each of W1and W2is belotecan.

4. A method for treating colorectal cancer (CRC) or non-small cell lung cancer (NSCLC) in a subject, the method comprising administering an ADC to the subject, wherein the ADC is represented by Formula (II):196NAI-5011166969vlwherein:Ab represents the antibody that binds to TF; ands is an integer from 1 to 10.

5. The method of any one of claims 1-4, wherein s is 2 or 4.

6. A method for treating colorectal cancer (CRC) or non-small cell lung cancer (NSCLC) in a subject, the method comprising administering an ADC to the subject, wherein the ADC is produced by conjugating an antibody that binds to TF (Ab) to a linker-payload of Formula (Ila):197NAI-5011166969vl(Ila).

7. The method of claim 6, wherein the ADC is produced by conjugating one Ab to one to ten, optionally, two or four, linker-payloads of Formula (Ila).

8. The method of any one of claims 1-7, wherein Ab comprises: (i) a VH CDR1, a VH CDR2, and a VH CDR3 as 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 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:26; or (ii) a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO:41 and a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO:42.

9. The method of any one of claims 1-8, wherein Ab comprises: a VH region comprising a VH CDR1 comprising the amino acid sequence of SEQ ID NOs:l, 7, 8, 15, 21, 27, 31, 32, 35, or 39, a VH CDR2 comprising the amino acid sequence of SEQ ID NOs:2, 9, 14, 16, or 22, and a VH CDR3 comprising the amino acid sequence of SEQ ID NOs:3, 10, 17, or 23; and a VL region comprising a VL CDR1 comprising the amino acid sequence of SEQ ID NOs:4, 11, 18, 24, 28, 33, 36, or 40, a VL CDR2198NAI-5011166969vlcomprising the amino acid sequence of SEQ ID NOs:5, 12, 19, 29, or 37, and a VL CDR3 comprising the amino acid sequence of SEQ ID NOs:6, 13, 20, 30, 34, or 38.

10. The method of any one of claims 1-9, wherein Ab comprises one or more of a framework 1 (FR1), a framework 2 (FR2), a framework 3 (FR3) and a framework 4 (FR4) sequence as set forth in any one of SEQ ID NOs:25, 26, 41, and 42.

11. The method of any one of claims 1-9, wherein Ab comprises human framework sequences.

12. The method of any one of claims 1-11, wherein Ab comprises a VH comprising the amino acid sequence of SEQ ID NO:25 and a VL comprising the amino acid sequence of SEQ IDNO:26.

13. The method of any one of claims 1-12, wherein the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:82.

14. The method of any one of claims 1-13, wherein the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:82 and a light chain comprising the amino acid sequence of SEQ ID NO:91.

15. The method of any one of claims 1-12, wherein the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:79.

16. The method of any one of claims 1-12 and 15, wherein the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:79 and a light chain comprising the amino acid sequence of SEQ ID NO:91.

17. The method of any one of claims 1-11, wherein Ab comprises: a VH comprising the amino acid sequence of SEQ ID NO:41 and a VL comprising the amino acid sequence of SEQ IDNO:42.

18. The method of any one of claims 1-11 and 17, wherein the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:85.

19. The method of any one of claims 1-11 and 17-18, wherein the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:85 and a light chain comprising the amino acid sequence of SEQ ID NO:92.199NAI-5011166969vl20. The method of any one of claims 1-11 and 17, wherein the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:88.

21. The method of any one of claims 1-11, 17 and 20, wherein the Ab comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:88 and a light chain comprising the amino acid sequence of SEQ ID NO:92.

22. A method for treating colorectal cancer (CRC) or non-small cell lung cancer (NSCLC) in a subject, the method comprising administering the subject a pharmaceutical composition comprising the ADC of any one of claims 1-21 and a pharmaceutically acceptable excipient.

23. The method of claim 22, wherein the pharmaceutical composition is characterized by an ADC drug-to-antibody ratio (DAR) of about 1 to about 20.

24. The method of claim 23, wherein the DAR is about 2 to about 8, optionally about 4 or about 8.

25. The method of any one of claims 1 to 24, wherein the cancer is characterized by expression of tissue factor.

26. The method of any one of claims 1-25, wherein the cancer is colorectal cancer.

27. The method of any one of claims claim 1-26, wherein the colorectal cancer is adenocarcinoma.

28. The method of claim 26 or 27, wherein the colorectal cancer is metastatic to lung.

29. The method of any one of claims 1-25, wherein the cancer is non-small cell lung cancer.

30. The method of any one of claims 1-25 and 29, wherein the non-small cell lung cancer is adenocarcinoma, squamous cell carcinoma, or large cell carcinoma.

31. The method of any one of claims 1-30, wherein the subject is human.200NAI-5011166969vl

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