Anti-Tissue Factor Antibodies, Antibody-Drug Conjugates, and Related Methods

Abstract

Provided herein are antibodies that specifically bind to human tissue factor (TF), anti-TF antibody-drug conjugates (ADCs), and compositions comprising the antibodies or ADCs. Also provided herein are methods of making and using the antibodies or ADCs, such as therapeutic and diagnostic methods.

1 - 503 . (canceled) 
     
     
         504 . An isolated human antibody which binds to the extracellular domain of human Tissue Factor (TF), wherein the antibody binds human TF at a human TF binding site that is distinct from a human TF binding site bound by human FVIIa. 
     
     
         505 . The isolated human antibody of  claim 504 , wherein
 (1) the isolated antibody does not inhibit human thrombin generation as determined by thrombin generation assay (TGA) optionally:   compared to a reference antibody comprising a VH sequence of SEQ ID NO:821 and a VL sequence of SEQ ID NO:822, or   wherein the antibody does not reduce the thrombin peak on a thrombin generation curve (Peak IIa) compared to an isotype control,   wherein the antibody does not increase the time from the assay start to the thrombin peak on a thrombin generation curve (ttPeak) compared to an isotype control, or wherein the antibody does not decrease the endogenous thrombin potential (ETP) as determined by the area under a thrombin generation curve compared to an isotype control;   
       and optionally wherein
 (2) the binding between the isolated antibody and a variant TF extracellular domain comprising a mutation at amino acid residue 149 of the sequence shown in SEQ ID NO:810 is less than 50% of the binding between the isolated antibody and the extracellular domain of TF of the sequence shown in SEQ ID NO:810, as determined by the median fluorescence intensity value of the isolated antibody relative to an isotype control in a live cell staining assay. 
 
     
     
         506 . The isolated human antibody of  claim 504 , wherein the antibody comprises:
 a VH-CDR1 comprising the sequence set forth in SEQ ID NO:779; a VH-CDR2 comprising the sequence set forth in SEQ ID NO:780; a VH-CDR3 comprising the sequence set forth in SEQ ID NO:781; a VL-CDR1 comprising the sequence set forth in SEQ ID NO:782; a VL-CDR2 comprising the sequence set forth in SEQ ID NO:783; and a VL-CDR3 comprising the sequence set forth in SEQ ID NO:784;   a VH-CDR1 comprising the sequence set forth in SEQ ID NO:872; a VH-CDR2 comprising the sequence set forth in SEQ ID NO:873; a VH-CDR3 comprising the sequence set forth in SEQ ID NO:874; a VL-CDR1 comprising the sequence set forth in SEQ ID NO:875; a VL-CDR2 comprising the sequence set forth in SEQ ID NO:876; and a VL-CDR3 comprising the sequence set forth in SEQ ID NO:877; or   a VH-CDR1 comprising the sequence set forth in SEQ ID NO:878; a VH-CDR2 comprising the sequence set forth in SEQ ID NO:879; a VH-CDR3 comprising the sequence set forth in SEQ ID NO:880; a VL-CDR1 comprising the sequence set forth in SEQ ID NO:881; a VL-CDR2 comprising the sequence set forth in SEQ ID NO:882; and a VL-CDR3 comprising the sequence set forth in SEQ ID NO:883.   
     
     
         507 . The isolated human antibody  claim 504 , wherein:
 the antibody binds human TF at a human TF binding site that is distinct from a human TF binding site bound by human FX; and/or   the antibody does not interfere with the ability of TF:FVIIa to convert FX into FXa; and/or   the antibody does not compete for binding to human TF with human FVIIa; and/or the antibody does not inhibit human thrombin generation as determined by thrombin generation assay (TGA), allows human thrombin generation as determined by thrombin generation assay (TGA), binds human TF at a human TF binding site that is distinct from a human TF binding site bound by human FX, does not interfere with the ability of TF:FVIIa to convert FX into FXa, and does not compete for binding to human TF with FVIIa; and/or   the antibody does not inhibit human thrombin generation as determined by thrombin generation assay (TGA), does not decrease the endogenous thrombin potential (ETP) as determined by the area under a thrombin generation curve compared to an isotype control, allows human thrombin generation as determined by thrombin generation assay (TGA), preserves the endogenous thrombin potential (ETP) as determined by the area under a thrombin generation curve compared to an isotype control, binds human TF at a human TF binding site that is distinct from a human TF binding site bound by human FX, does not interfere with the ability of TF:FVIIa to convert FX into FXa, and does not compete for binding to human TF with FVIIa; and/or   the antibody does not inhibit human thrombin generation as determined by thrombin generation assay (TGA), does not reduce the thrombin peak on a thrombin generation curve (Peak IIa) compared to an isotype control, does not increase the time from the assay start to the thrombin peak on a thrombin generation curve (ttPeak) compared to an isotype control, does not decrease the endogenous thrombin potential (ETP) as determined by the area under a thrombin generation curve compared to an isotype control, allows human thrombin generation as determined by thrombin generation assay (TGA), maintains the thrombin peak on a thrombin generation curve (Peak IIa) compared to an isotype control, maintains the time from the assay start to the thrombin peak on a thrombin generation curve (ttPeak) compared to an isotype control, preserves the endogenous thrombin potential (ETP) as determined by the area under a thrombin generation curve compared to an isotype control, binds human TF at a human TF binding site that is distinct from a human TF binding site bound by human FX, does not interfere with the ability of TF:FVIIa to convert FX into FXa, and does not compete for binding to human TF with FVIIa; and/or   the antibody inhibits FVIIa-dependent TF signaling; and/or   
       the binding between the isolated antibody and a variant TF extracellular domain comprising a mutation at amino acid residue 149, and/or or at amino acid residues 171 and 197, of the sequence shown in SEQ ID NO:810 is less than 50% of the binding between the isolated antibody and the extracellular domain of TF of the sequence shown in SEQ ID NO:810, as determined by the median fluorescence intensity value of the isolated antibody relative to an isotype control in a live cell staining assay, optionally wherein the mutation at amino acid residue 149 of the sequence shown in SEQ ID NO:810 is K149N, and/or or the mutations at amino acid residues 171 and 197 of the sequence shown in SEQ ID NO:810 are N171H and T197K; and/or
 the binding between the isolated antibody and a variant TF extracellular domain comprising a mutation at amino acid residue 68 of the sequence shown in SEQ ID NO:810 is greater than 50% of the binding between the isolated antibody and the extracellular domain of TF of the sequence shown in SEQ ID NO:810, as determined by the median fluorescence intensity value of the isolated antibody relative to an isotype control in a live cell staining assay, optionally wherein the mutation at amino acid residue 68 of the sequence shown in SEQ ID NO:810 is K68N; and/or 
 the binding between the isolated antibody and a human TF extracellular domain with amino acid residues (i) 1-77, (ii) 39-77, or (iii) 94-107 of the sequence shown in SEQ ID NO:810 replaced by rat TF extracellular domain amino acid residues (i) 1-76, (ii) 39-77, or (iii) 99-112, respectively of the sequence shown in SEQ ID NO:838 is greater than 50% of the binding between the isolated antibody and the extracellular domain of TF of the sequence shown in SEQ ID NO:810, as determined by the median fluorescence intensity value of the isolated antibody relative to an isotype control in a live cell staining assay; and/or 
 the binding between the isolated antibody and a human TF extracellular domain with amino acid residues (i) 146-158, (ii) 159-219, (iii) 159-289, (iv) 159-174, or (v) 167-174 of the sequence shown in SEQ ID NO:810 replaced by rat TF extracellular domain amino acid residues (i) 151-163, (ii) 164-224, (iii) 164-194, (iv) 164-179, or (v) 172-179, respectively of the sequence shown in SEQ ID NO:838 is less than 50% of the binding between the isolated antibody and the extracellular domain of TF of the sequence shown in SEQ ID NO:810, as determined by the median fluorescence intensity value of the isolated antibody relative to an isotype control in a live cell staining assay; and/or 
 the binding between the isolated antibody and a rat TF extracellular domain with amino acid residues 141-194 of the sequence shown in SEQ ID NO:838 replaced by human TF extracellular domain amino acid residues 136-189 of the sequence shown in SEQ ID NO:810 is greater than 50% of the binding between the isolated antibody and the extracellular domain of TF of the sequence shown in SEQ ID NO:810, as determined by the median fluorescence intensity value of the isolated antibody relative to an isotype control in a live cell staining assay. 
 
     
     
         508 . The isolated human antibody of  claim 504 , wherein:
 the antibody binds to cynomolgus, mouse, rabbit, and/or pig TF; and/or
 the antibody reduces lesion size in a swine choroidal neovascularization (CNV) model; and/or 
 the antibody: 
 (a) does not inhibit human thrombin generation as determined by thrombin generation assay (TGA); and 
 (b) the binding between the antibody and a variant TF extracellular domain comprising mutations at amino acid residues 171 and 197, optionally N171H and T197K, of the sequence shown in SEQ ID NO:810 is less than 50% of the binding between the antibody and the extracellular domain of TF of the sequence shown in SEQ ID NO:810, as determined by the median fluorescence intensity value of the antibody relative to an isotype control in a live cell staining assay. 
   
     
     
         509 . An isolated antibody that competes for binding to human TF with, or binds the human TF epitope bound by, an isolated human antibody which binds to the extracellular domain of human Tissue Factor (TF), wherein the antibody binds human TF at a human TF binding site that is distinct from a human TF binding site bound by human FVIIa. 
     
     
         510 . The isolated human antibody of  claim 504 , wherein the antibody competes for binding to human TF with, binds to the same human TF epitope bound by, comprises all three heavy chain Complementary Determining Regions (CDRs) and all three light chain CDRs from, comprises, or consists of: the antibody designated 25A, the antibody designated 25A3, the antibody designated 25A5, the antibody designated 25A5-T, the antibody designated 25G, the antibody designated 25G1, the antibody designated 25G9, the antibody designated 43B, the antibody designated 43B1, the antibody designated 43B7, the antibody designated 43D, the antibody designated 43D7, the antibody designated 43D8, the antibody designated 43E, or the antibody designated 43Ea,
 optionally wherein   the three heavy chain CDRs and the three light chain CDRs are determined using Kabat, Chothia, AbM, Contact, or IMGT numbering; and/or   the antibody is human, humanized, or chimeric; and/or   the antibody comprises a VH sequence of SEQ ID NO:113 and a VL sequence of SEQ ID NO:114; and/or   the antibody comprises a VH sequence of SEQ ID NO:151 and a VL sequence of SEQ ID NO:152; and/or   the antibody comprises a VH sequence of SEQ ID NO:189 and a VL sequence of SEQ ID NO:190; and/or   the antibody comprises a VH sequence of SEQ ID NO:836 and a VL sequence of SEQ ID NO:837; and/or   the antibody comprises a VH sequence of SEQ ID NO:227 and a VL sequence of SEQ ID NO:228; and/or   the antibody comprises a VH sequence of SEQ ID NO:265 and a VL sequence of SEQ ID NO:266; and/or   the antibody comprises a VH sequence of SEQ ID NO:303 and a VL sequence of SEQ ID NO:304; and/or   the antibody comprises a VH sequence of SEQ ID NO:455 and a VL sequence of SEQ ID NO:456; and/or   the antibody comprises a VH sequence of SEQ ID NO:493 and a VL sequence of SEQ ID NO:494; and/or   the antibody comprises a VH sequence of SEQ ID NO:531 and a VL sequence of SEQ ID NO:532; and/or   the antibody comprises a VH sequence of SEQ ID NO:569 and a VL sequence of SEQ ID NO:570; and/or   the antibody comprises a VH sequence of SEQ ID NO:607 and a VL sequence of SEQ ID NO:608; and/or   the antibody comprises a VH sequence of SEQ ID NO:645 and a VL sequence of SEQ ID NO:646; and/or   the antibody comprises a VH sequence of SEQ ID NO:683 and a VL sequence of SEQ ID NO:684; and/or   the antibody comprises a VH sequence of SEQ ID NO:721 and a VL sequence of SEQ ID NO:722; and/or   the antibody comprises: a VH-CDR1 comprising the sequence set forth in SEQ ID NO:779; a VH-CDR2 comprising the sequence set forth in SEQ ID NO:780; a VH-CDR3 comprising the sequence set forth in SEQ ID NO:781; a VL-CDR1 comprising the sequence set forth in SEQ ID NO:782; a VL-CDR2 comprising the sequence set forth in SEQ ID NO:783; and a VL-CDR3 comprising the sequence set forth in SEQ ID NO:784; and/or   
       the antibody comprises: a VH-CDR1 comprising the sequence set forth in SEQ ID NO:872; a VH-CDR2 comprising the sequence set forth in SEQ ID NO:873; a VH-CDR3 comprising the sequence set forth in SEQ ID NO:874; a VL-CDR1 comprising the sequence set forth in SEQ ID NO:875; a VL-CDR2 comprising the sequence set forth in SEQ ID NO:876; and a VL-CDR3 comprising the sequence set forth in SEQ ID NO:877; and/or
 the antibody comprises: a VH-CDR1 comprising the sequence set forth in SEQ ID NO:878; a VH-CDR2 comprising the sequence set forth in SEQ ID NO:879; a VH-CDR3 comprising the sequence set forth in SEQ ID NO:880; a VL-CDR1 comprising the sequence set forth in SEQ ID NO:881; a VL-CDR2 comprising the sequence set forth in SEQ ID NO:882; and a VL-CDR3 comprising the sequence set forth in SEQ ID NO:883; and/or 
 
       the antibody comprises: a VH-CDR1 comprising the sequence set forth in SEQ ID NO:797; a VH-CDR2 comprising the sequence set forth in SEQ ID NO:798; a VH-CDR3 comprising the sequence set forth in SEQ ID NO:799; a VL-CDR1 comprising the sequence set forth in SEQ ID NO:800; a VL-CDR2 comprising the sequence set forth in SEQ ID NO:801; and a VL-CDR3 comprising the sequence set forth in SEQ ID NO:802; and/or
 the antibody comprises a VH sequence of SEQ ID NO:763 and a VL sequence of SEQ ID NO:764; and/or 
 the antibody comprises a VH sequence of SEQ ID NO:868 and a VL sequence of SEQ ID NO:869; and/or 
 
       the antibody comprises a VH sequence of SEQ ID NO:870 and a VL sequence of SEQ ID NO:871; and/or
 the antibody comprises a VH sequence of SEQ ID NO:769 and a VL sequence of SEQ ID NO:770. 
 
     
     
         511 . An isolated antibody that competes for binding to human TF with: the antibody designated 1F, the antibody designated 1G, the antibody designated 29D, the antibody designated 29E, the antibody designated 39A, or the antibody designated 54E. 
     
     
         512 . The isolated antibody of  claim 511 , comprising all three heavy chain Complementary Determining Regions (CDRs) and all three light chain CDRs from, or comprising, or consisting of: the antibody designated 1F, the antibody designated 1G, the antibody designated 29D, the antibody designated 29E, the antibody designated 39A, the antibody designated 43Ea, or the antibody designated 54E, optionally wherein the three heavy chain CDRs and the three light chain CDRs are determined using Kabat, Chothia, AbM, Contact, or IMGT numbering; and/or
 the antibody comprises all three heavy chain CDRs and all three light chain CDRs from the antibody designated 1F, 1G, 29D, 29E, 39A, or 54E; or   the antibody comprises a VH sequence of SEQ ID NO:37 and a VL sequence of SEQ ID NO:38; or   the antibody comprises a VH sequence of SEQ ID NO:75 and a VL sequence of SEQ ID NO:76; or   the antibody comprises a VH sequence of SEQ ID NO:341 and a VL sequence of SEQ ID NO:342; or   the antibody comprises a VH sequence of SEQ ID NO:379 and a VL sequence of SEQ ID NO:380; or   the antibody comprises a VH sequence of SEQ ID NO:417 and a VL sequence of SEQ ID NO:418; or   the antibody comprises a VH sequence of SEQ ID NO:759 and a VL sequence of SEQ ID NO:760; or   the antibody comprises: a VH-CDR1 comprising the sequence set forth in SEQ ID NO:773; a VH-CDR2 comprising the sequence set forth in SEQ ID NO:774; a VH-CDR3 comprising the sequence set forth in SEQ ID NO:775; a VL-CDR1 comprising the sequence set forth in SEQ ID NO:776; a VL-CDR2 comprising the sequence set forth in SEQ ID NO:777; and a VL-CDR3 comprising the sequence set forth in SEQ ID NO:778; or   the antibody comprises: a VH-CDR1 comprising the sequence set forth in SEQ ID NO:785; a VH-CDR2 comprising the sequence set forth in SEQ ID NO:786; a VH-CDR3 comprising the sequence set forth in SEQ ID NO:787; a VL-CDR1 comprising the sequence set forth in SEQ ID NO:788; a VL-CDR2 comprising the sequence set forth in SEQ ID NO:789; and a VL-CDR3 comprising the sequence set forth in SEQ ID NO:790; or   the antibody comprises: a VH-CDR1 comprising the sequence set forth in SEQ ID NO:791; a VH-CDR2 comprising the sequence set forth in SEQ ID NO:792; a VH-CDR3 comprising the sequence set forth in SEQ ID NO:793; a VL-CDR1 comprising the sequence set forth in SEQ ID NO:794; a VL-CDR2 comprising the sequence set forth in SEQ ID NO:795; and a VL-CDR3 comprising the sequence set forth in SEQ ID NO:796; or   the antibody comprises: a VH-CDR1 comprising the sequence set forth in SEQ ID NO:803; a VH-CDR2 comprising the sequence set forth in SEQ ID NO:804; a VH-CDR3 comprising the sequence set forth in SEQ ID NO:805; a VL-CDR1 comprising the sequence set forth in SEQ ID NO:806; a VL-CDR2 comprising the sequence set forth in SEQ ID NO:807; and a VL-CDR3 comprising the sequence set forth in SEQ ID NO:808; or   the antibody comprises a VH sequence of SEQ ID NO:761 and a VL sequence of SEQ ID NO:762; or   the antibody comprises a VH sequence of SEQ ID NO:765 and a VL sequence of SEQ ID NO:766; or   the antibody comprises a VH sequence of SEQ ID NO:767 and a VL sequence of SEQ ID NO:768; or   the antibody comprises a VH sequence of SEQ ID NO:771 and a VL sequence of SEQ ID NO:772.   
     
     
         513 . An isolated polynucleotide or set of polynucleotides encoding the antibody of  claim 504 , a VH thereof, a VL thereof, a light chain thereof, a heavy chain thereof, or an antigen-binding portion thereof. 
     
     
         514 . A vector or set of vectors comprising the polynucleotide or set of polynucleotides of  claim 513 . 
     
     
         515 . A host cell comprising the polynucleotide or set of polynucleotides of  claim 513 . 
     
     
         516 . A method of producing an antibody comprising expressing the antibody with the host cell of  claim 515  and isolating the expressed antibody. 
     
     
         517 . An antibody-drug conjugate comprising: an anti-human Tissue Factor (anti-hTF) antibody, a cytotoxic agent linked to the antibody, and optionally a linker that links the antibody to the cytotoxic agent, wherein the antibody binds to the extracellular domain of human Tissue Factor (TF) at a human TF binding site that is distinct from a human TF binding site bound by human FVIIa. 
     
     
         518 . A pharmaceutical composition comprising the antibody of  claim 504  and a pharmaceutically acceptable excipient. 
     
     
         519 . A method of treating or preventing a disease or condition in a subject in need thereof, comprising administering to the subject an effective amount of an antibody of  claim 504   
     
     
         520 . The method of  claim 519 , wherein the disease or condition is cancer, optionally head and neck cancer, ovarian cancer, gastric cancer, esophageal cancer, cervical cancer, prostate cancer, pancreatic cancer, estrogen receptors negative (ER−), progesterone receptors negative (PR−), and HER2 negative (HER2−) triple negative breast cancer, glioblastoma, lung cancer, bladder cancer, melanoma or kidney cancer;
 or wherein the method is for treatment of a disease or condition that involves neovascularization, optionally age-related macular degeneration (AMD), diabetic retinopathy or cancer, and optionally wherein the disease or condition involves vascular inflammation. 
 
     
     
         521 . The method of  claim 519 , further comprising administering one or more additional therapeutic agents to the subject, optionally wherein
 the additional therapeutic agent is formulated in the same pharmaceutical composition as the antibody or antibody-drug conjugate, or   wherein the additional therapeutic agent is formulated in a different pharmaceutical composition from the antibody or antibody-drug conjugate, or   wherein the additional therapeutic agent is administered prior to administering the antibody or antibody-drug conjugate, or   wherein the additional therapeutic agent is administered after administering the antibody or antibody-drug conjugate, or   wherein the additional therapeutic agent is administered contemporaneously with the antibody or antibody-drug conjugate.   
     
     
         522 . A method of detecting TF in a subject having or suspected of having a disease or condition, the method comprising:
 (i) (a) administering to the subject the antibody of  claim 504 ; and (b) detecting the presence or the level of TF in the subject;   
       or
 (ii) (a) receiving a sample from the subject; and (b) detecting the presence or the level of TF in the sample by contacting the sample with the antibody of  claim 504 . 
 
     
     
         523 . A kit comprising
 (i) the antibody of  claim 504 ; and   (ii) instructions for use.