EP4583922A2 - Conjugués anticorps-médicament de facteur tissulaire et leurs utilisations - Google Patents

Conjugués anticorps-médicament de facteur tissulaire et leurs utilisations

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Publication number
EP4583922A2
EP4583922A2 EP23863945.4A EP23863945A EP4583922A2 EP 4583922 A2 EP4583922 A2 EP 4583922A2 EP 23863945 A EP23863945 A EP 23863945A EP 4583922 A2 EP4583922 A2 EP 4583922A2
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EP
European Patent Office
Prior art keywords
substituted
alkyl
amino acid
aryl
amino
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP23863945.4A
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German (de)
English (en)
Inventor
Robyn M. BARFIELD
Maxine Bauzon
Penelope M. DRAKE
Seema Kantak
Brian Alan MENDELSOHN
Tiffany UNSULANGI
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Exelixis Inc
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Exelixis Inc
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Filing date
Publication date
Application filed by Exelixis Inc filed Critical Exelixis Inc
Publication of EP4583922A2 publication Critical patent/EP4583922A2/fr
Pending legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6849—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A61K47/68037—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a camptothecin [CPT] or derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6889—Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/36—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against blood coagulation factors

Definitions

  • the present disclosure relates generally to antibody-drug conjugates (ADCs) that bind to tissue factor (TF, e.g., human TF) and methods of their use.
  • ADCs antibody-drug conjugates
  • the Hydrazino-Ao-Pictet-Spengler (HIPS) conjugation method 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.
  • FGE formyl generating enzyme
  • 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.
  • ADCs that can target TF to treat, prevent, or alleviate TF-mediated diseases, disorders, or conditions, such as those involving tumor cells expressing TF.
  • the present disclosure also provides methods of treating, preventing, or alleviating a TF-mediated disease, disorder, or condition, such as alleviating one or more symptoms of the TF-mediated disease, disorder, or condition with a TF-ADC.
  • a TF-ADC comprising (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-Ao-Pictet-Spengler (HIPS) conjugation method.
  • HIPS Hydrazino-Ao-Pictet-Spengler
  • 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).
  • 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.
  • 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.
  • a TF-ADC is represented by Formula (I), the TF-ADC comprising: a. an antibody that binds to tissue factor (TF); and b. two or more drugs conjugated to a pyridazine-pyrrolo coupling moiety, each via a linker wherein:
  • Ab represents the antibody that binds to TF
  • Z 1 , Z 2 , and Z 4 are each independently CR 4 ;
  • R 1 , R 2 , R 3 , and R 4 are each selected from hydrogen and alkyl
  • L A is a first linker comprising:
  • L B is a second linker comprising:
  • T 7 , T 8 , T 9 , T 10 , T 11 , T 12 and T 13 are 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 , -(CR 13 OH) X -, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para- amino-benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxy
  • the glycoside is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.
  • T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , and T 12 are each optionally substituted with a glycoside.
  • MABO, MABC, PABO, PABC, PAB, PABA, PAP and PHP are each optionally substituted with a glycoside.
  • T 8 is (Ci-Ci 2 )alkyl and V 8 is -CONH-;
  • 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.
  • a glycoside such as a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.
  • Z 1 , Z 2 , and Z 4 are each independently CR 4 ;
  • L A is a first linker wherein:
  • T 1 is (Ci-Ci2)alkyl and V 1 is -C0NH-;
  • T 2 is substituted (Ci-Ci2)alkyl and V 2 is -CO-;
  • T 3 is (AA) P where p is an integer from 1-20 and V 3 is a covalent bond;
  • T 4 is PABC and V 4 is a covalent bond; a, b, c, and d are each 1; e and f are each 0; and
  • T 7 is a covalent bond and V 7 is -NHCO-;
  • T 8 is (Ci-Ci 2 )alkyl and V 8 is -CONH-;
  • T 9 is substituted (Ci-Ci2)alkyl and V 9 is -CO-;
  • T 10 is (AA) P where p is an integer from 1-20 and V 10 is a covalent bond;
  • T 11 is PABC and V 11 is a covalent bond; and g, h, i, j, and k are each 1; and
  • 1 and m are each 0; s is an integer from 1 to 10;
  • W 1 is a first drug
  • W 2 is a second drug.
  • W 1 and W 2 are camptothecin analogues, for example, belotecan.
  • a TF-ADC is represented by Formula (I): wherein:
  • Ab represents the antibody that binds to TF
  • Z 1 , Z 2 , and Z 4 are each independently CR 4 ;
  • Z 3 is C-L B -W 2 ;
  • R 1 , R 2 , R 3 and R 4 are each selected from hydrogen and (Ci-Ci2)alkyl;
  • L A is a linker wherein:
  • T 1 is (Ci-Ce)alkyl and V 1 is -CONH-;
  • T 2 is (Ci-Ce)alkylene substituted with -NHCO(PEG)k, wherein k is an integer from 2 to 10 and V 2 is -CO-;
  • T 7 is a covalent bond and V 7 is -NHCO-;
  • Ab represents the antibody that binds to TF; and s is an integer from 1 to 10.
  • Formula (II) may be prepared by conjugating one or more linker-payloads of Formula (Ila), shown below, with a TF antibody:
  • a TF-ADC can be represented by Formula (I) or (II) 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.
  • FIG. 9A shows a graph of mean tumor volume (mm 3 ) vs. days, which indicates in vivo efficacy against a BxPC3 xenograft of TF -targeted ADCs.
  • FIG. 9B shows body weights of the tested mice.
  • FIG. 9C plots the in vivo efficacy data of TF-ADC 6-8 and TF-ADC 7-8.
  • FIG. 9D plots the in vivo efficacy data of TF-ADC 6-4 and TF-ADC 7-4.
  • FIG. 9E plots the in vivo efficacy data of TF-ADC 6-4 and TF-ADC 6-8.
  • FIG. 9F plots the in vivo efficacy data of TF-ADC 7-4 and TF-ADC 7-8.
  • a single i.v. dose was delivered on Day 0.
  • FIG. 16 plots HMGB1 released by A431 tumor cells treated with control, 33 nM TF- ADC 6-8, 33 nM belotecan, 100 nM MMAE, or 100 nM benchmark ADC.
  • 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.
  • 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.
  • groups include, without limitation, fluoroalkyl groups, such as trifluoromethyl, difluoromethyl, trifluoroethyl and the like.
  • 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.
  • 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.
  • 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, thio
  • Heteroaryloxy refers to -O-heteroaryl.
  • 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.
  • 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.
  • 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,
  • heterocyclylthio refers to the group heterocyclic-S-.
  • heterocyclene refers to the diradical group formed from a heterocycle, as defined herein.
  • hydroxyamino refers to the group -NHOH.
  • Niro refers to the group -NO2.
  • Thiol refers to the group -SH.
  • Alkylthio or the term “thioalkoxy” refers to the group -S-alkyl, wherein alkyl is as defined herein.
  • sulfur may be oxidized to -S(O)-.
  • the sulfoxide may exist as one or more stereoisomers.
  • thioaryloxy refers to the group aryl-S- wherein the aryl group is as defined herein including optionally substituted aryl groups also defined herein.
  • heteroaryl oxy refers to the group heteroaryl-S- wherein the heteroaryl group is as defined herein including optionally substituted aryl groups as also defined herein.
  • heterocyclooxy refers to the group heterocyclyl-S- wherein the heterocyclyl group is as defined herein including optionally substituted heterocyclyl groups as also defined herein.
  • 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.
  • substituent groups for substituting for one or more hydrogens are, unless otherwise specified, -R 60 , halo,
  • -NR 80 R 80 is meant to include -NH2, -NH-alkyl, 7V-pyrrolidinyl, 7V-piperazinyl, 47V-methyl-piperazin-l-yl and A-morpholinyl.
  • substituent groups for hydrogens on unsaturated carbon atoms in “substituted” alkene, alkyne, aryl and heteroaryl groups are, unless otherwise specified, -R 60 , halo, -O M + , -OR 70 , -SR 70 , -S’M + , -NR 80 R 80 , trihalomethyl, -CF3, -CN, -OCN, -SCN, -NO, -NO2, -N3, -SO2R 70 , -SO 3 M + , -SO3R 70 , -OSO2R 70 , -OSO3-M+ -OSO3R 70 , -PO3' 2 (M + )2, -P(O)(OR 70 )O-M + , -P(O)(OR 70 )2, -C(O)R 70 , -C(S)R 70 , -C(NR 70
  • substituent groups for hydrogens on nitrogen atoms in “substituted” heteroalkyl and cycloheteroalkyl groups are, unless otherwise specified, -R 60 , -O M + , -OR 70 , -SR 70 , -S'M + , -NR 80 R 80 , trihalomethyl, -CF3, -CN, -NO, -NO2, -S(O) 2 R 70 , -S(O) 2 O'M + , -S(O) 2 OR 70 , -OS(O) 2 R 70 , -OS(O) 2 O-M + , -OS(O) 2 OR 70 , -P(O)(O-) 2 (M + )2, -P(O)(OR 70 )O'M + , -P(O)(OR 70 )(OR 70 ), -C(O) R 70 , -C
  • pyrazoles imidazoles, benzimidazoles, triazoles, and tetrazoles.
  • “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.
  • 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.
  • “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.
  • “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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • CDRs complementarity determining regions
  • 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.
  • the antibodies produced by the antibody producing cells isolated from a first animal immunized with an antigen are natural antibodies.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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 preexisting amino acid sequence modifications (such as additions, deletions and/or substitutions).
  • 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.
  • 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.
  • 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.
  • 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.
  • FGE Formylglycine Generating Enzyme
  • 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.
  • a converted aldehyde tag e.g., a reactive aldehyde group
  • aldehyde-reactive reactive partner which comprises an aldehyde-reactive group and a moiety of interest
  • 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.
  • subject refers to human and non-human subjects, especially mammalian subjects.
  • treating means the treating or treatment of a disease or medical condition in a subject, 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 subject; (c) suppressing the disease or medical condition, for example by, slowing or arresting the development of the disease or medical condition in a subject; or (d) alleviating a symptom of the disease or medical condition in a subject.
  • the term “treating,” or “treatment” excludes a prophylactic treatment.
  • amino acid sequence refers to the amino acid sequence of a polypeptide prior to modification to include a modified amino acid residue.
  • amino acid analog 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).
  • 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
  • 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 embodiments, 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 can 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, and the like) or an atom (such as Cl or Br, and the like), deletion of a group, substitution of a covalent bond (single bond for double bond, and the like), or combinations thereof.
  • amino acid analogs can include a-hydroxy acids, and a- amino acids, and the like.
  • amino acid side chain is 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.
  • carbohydrate is used to refer to monomer units and/or polymers of monosaccharides, disaccharides, oligosaccharides, and polysaccharides.
  • sugar is be used to refer to the smaller carbohydrates, such as monosaccharides, disaccharides.
  • tissue Factor tissue Factor
  • platelet tissue factor factor III
  • thromboplastin thromboplastin
  • CD 142 tissue Factor
  • the human TF does not comprise a signal peptide, for example amino acid (aa) 33 to aa 295 of SEQ ID NO: 175.
  • 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.
  • the TF protein is cynomolgus TF (cTF;
  • 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.
  • 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).
  • RIAs radioimmunoassays
  • ELISAs enzyme linked immunosorbent assays
  • 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.
  • assays examples 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.
  • RIA solid phase direct or indirect radioimmunoassay
  • EIA enzyme immunoassay
  • 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.
  • 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).
  • a purified antigen e.g., TF, such as human TF
  • test antigen binding protein e.g., test TF antibody or ADC
  • a labeled reference antigen binding protein e.g., reference TF antibody or ADC
  • 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).
  • 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).
  • a TF antibody can react with TF sequences other than human TF sequences (e.g., cynomolgus TF sequences).
  • variable 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 binding 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.
  • the variable region is a human variable region.
  • hypervariable region refers to the regions of an antibody variable region that are hypervariable in sequence and/or form structurally defined loops.
  • 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).
  • 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.
  • IMGT® ImMunoGeneTics
  • IG immunoglobulins
  • TR T cell receptors
  • MHC major histocompatibility complex
  • 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.
  • hypervariable region As used herein, the terms “hypervariable region,” “HVR,” “HV,” “complementarity determining region,” or “CDR” are used interchangeably.
  • vector refers to a substance that is used to carry or include a nucleic acid sequence, for example, to introduce a nucleic acid sequence into a host cell.
  • vectors include expression vectors, plasmids, phage vectors, viral vectors, episomes and artificial chromosomes, which can include selection sequences or markers operable for stable integration into a host cell’s chromosome.
  • a vector can include one or more selectable marker genes and appropriate expression control sequences. Selectable marker genes that can be included, for example, provide resistance to antibiotics or toxins, complement auxotrophic deficiencies, or supply critical nutrients not in the culture media.
  • nucleic acid analysis such as Northern blots or polymerase chain reaction (PCR) amplification of mRNA, or immunoblotting for expression of gene products, or other suitable analytical methods to test the expression of an introduced nucleic acid sequence or its corresponding gene product.
  • PCR polymerase chain reaction
  • suitable analytical methods to test the expression of an introduced nucleic acid sequence or its corresponding gene product.
  • the nucleic acid molecules are expressed in a sufficient amount to produce a desired product (e.g., a TF antibody), and it is further understood that expression levels can be optimized to obtain sufficient expression using methods well known in the art.
  • TF-mediated disease includes a cancer including, but not limited to, cancers that express or overexpress TF.
  • 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.
  • drugs include small molecule drugs, such as a cancer chemotherapeutic agent.
  • a cancer chemotherapeutic agent such as 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 (z.e., 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.
  • 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 Whyzodiazepine (PBD)).
  • PBD pyrroleauzodiazepine
  • 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 (CYNOTANTM), melphalan (L-sarcolysin), carmustine (BCNU), lomustine (CCNU), semustine (methyl-CCNU), streptozocin, chlorozotocin, uracil mustard, chlormethine, ifosfamide, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, dacarbazine, and temozolomide.
  • alkylating agents such as nitrogen mustards, nitrosoureas, ethylenimine derivatives, alkyl sulfonates, and triazenes,
  • Antimetabolite agents include folic acid analogs, pyrimidine analogs, purine analogs, and adenosine deaminase inhibitors, including, but not limited to, cytarabine (CYTOSAR-U®), cytosine arabinoside, fluorouracil (5-FU), floxuridine (FudR), 6- thioguanine, 6-mercaptopurine (6-MP), pentostatin, 5 -fluorouracil (5-FU), methotrexate, 10- propargyl-5,8-dideazafolate (PDDF, CB3717), 5,8-dideazatetrahydrofolic acid (DDATHF), leucovorin, fludarabine phosphate, pentostatin, and gemcitabine.
  • CYTOSAR-U® cytosine arabinoside
  • fluorouracil (5-FU) floxuridine
  • 6-MP 6-mercaptopurine
  • pentostatin 5 -fluorouraci
  • Suitable natural products and their derivatives include, but are not limited to, Ara-C, paclitaxel (TAXOL®), docetaxel (TAXOTERE®), deoxycoformycin, mitomycin-C, L- asparaginase, azathioprine; brequinar; alkaloids, e.g. vincristine, vinblastine, vinorelbine, vindesine, 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.
  • cytotoxic agents are navelbene, CPT-11, anastrazole, letrazole, capecitabine, reloxafine, cyclophosphamide, ifosamide, and droloxafine.
  • 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.
  • Hormone modulators and steroids 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.
  • chemotherapeutic agents include metal complexes, e.g. cisplatin (cis- DDP), carboplatin, and the like; ureas, e.g. hydroxyurea; hydrazines, e.g. N-methylhydrazine; epidophyllotoxin; a topoisomerase inhibitor; procarbazine; mitoxantrone; leucovorin; tegafur; and the like
  • metal complexes e.g. cisplatin (cis- DDP), carboplatin, and the like
  • ureas e.g. hydroxyurea
  • hydrazines e.g. N-methylhydrazine
  • epidophyllotoxin e.g. N-methylhydrazine
  • a topoisomerase inhibitor e.g. N-methylhydrazine
  • procarbazine mitoxantrone
  • leucovorin tegafur
  • mycophenolic acid 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.
  • 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.
  • 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).
  • analogs and derivatives e.g., TAXOTERE® docetaxel, as noted herein
  • paclitaxel conjugates e.g., paclitaxel -PEG, paclitaxel-dextran, or paclitaxel -xylose.
  • Taxane 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.
  • Biological response modifiers suitable for use include, but are not limited to, (1) inhibitors of tyrosine kinase (RTK) activity; (2) inhibitors of serine/threonine kinase activity; (3) tumor-associated antigen antagonists, such as antibodies that 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.
  • RTK tyrosine kinase
  • 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.
  • therapeutically effective amount 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).
  • 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 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.
  • 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.
  • pharmaceutically acceptable 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.
  • 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.
  • 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.
  • 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 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.
  • TF antibody also referred to herein as “TF antibody,” “anti-TF antibody,” “TF Ab,” “Ab” or “antibody”
  • 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.
  • 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.
  • the one or more functional groups and covalent bonds can include a branched linker as described herein.
  • Moi eties of interest can be conjugated to the TF antibody at any desired site of the antibody.
  • 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.
  • 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.
  • the TF antibody and the moi eties of interest are conjugated through a conjugation moiety.
  • 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.
  • 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.
  • 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.
  • a conjugation moiety e.g., a hydrazinyl-indolyl or a hydrazinyl- pyrrolo-pyridinyl conjugation moiety
  • R is attached to two or more drugs or active agents, R.
  • Combinations of the same or different payloads may be conjugated to the TF antibody through the branched linker.
  • 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).
  • 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.
  • the drugs or active agents may be selected from drugs and active agents that have a synergistic therapeutic effect.
  • synergistic 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.
  • 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.
  • linker is a cleavable linker, such as a cleavable linker as described herein.
  • 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 conjugation 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.
  • 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).
  • 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”).
  • FGE formylglycine generating enzyme
  • the fGly residue generated by an FGE may also be referred to as a “formylglycine”.
  • aldehyde tag is used herein to refer to an amino acid sequence that includes a “converted” sulfatase motif (z.e., 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).
  • 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 (i.e., 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).
  • an “unconverted” sulfatase motif i.e., 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).
  • 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.
  • 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).
  • a compound e.g., a compound containing a hydrazinyl-indolyl or a hydrazinyl-pyrrolo- pyridinyl conjugation moiety, as described above.
  • 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.
  • the reactive partner may include a hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety as described above.
  • two or more drugs or active agents may be attached to a hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety.
  • 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.
  • 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).
  • 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.
  • an amino acid residue of the TF antibody is a cysteine or serine residue that is modified to an fGly residue, as described above.
  • 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.
  • 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).
  • 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.
  • 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.
  • Ab represents the antibody that binds to TF
  • Z 1 , Z 2 , Z 3 and Z 4 are each independently selected from CR 4 , N and C-L B -W 2 , wherein at least one Z 1 , Z 2 , Z 3 and Z 4 is C-L B -W 2 ;
  • R 1 is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl;
  • R 2 and R 3 are each independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, 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 R 2 and R 3 are optionally cyclically linked to form a 5 or 6-membered heterocyclyl; each R 4 is independently selected from hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl
  • L A is a first linker
  • L B is a second linker; s is an integer from 1 to 10;
  • W 1 is a first drug
  • W 2 is a second drug.
  • Z 1 , Z 2 , Z 3 and Z 4 are each independently selected from CR 4 , N and C-L B -W 2 , wherein at least one Z 1 , Z 2 , Z 3 and Z 4 is C-L B -W 2 .
  • Z 1 is CR 4 .
  • Z 1 is N.
  • Z 1 is C- L B -W 2 .
  • Z 2 is CR 4 .
  • Z 2 is N.
  • Z 2 is C-L B -W 2 .
  • Z 3 is CR 4 .
  • Z 3 is N.
  • Z 1 is C-L B -W 2
  • Z 2 is CR 4
  • Z 3 is CR 4
  • Z 4 is CR 4
  • Z 1 is CR 4
  • Z 2 is C-L B -W 2
  • Z 3 is CR 4
  • Z 4 is CR 4
  • Z 1 is CR 4
  • Z 2 is CR 4
  • Z 3 is C-L B - W 2
  • Z 4 is CR 4 .
  • Z 1 is CR 4
  • Z 2 is CR 4
  • Z 3 is CR 4
  • Z 4 is C-L B -W 2 .
  • R 1 is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, substituted heterocyclyl.
  • R 1 is hydrogen.
  • R 1 is 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.
  • R 1 is 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.
  • R 1 is 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.
  • R 1 is 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.
  • R 1 is 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.
  • R 1 is 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.
  • R 1 is 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.
  • R 2 and R 3 are each independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, 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 R 2 and R 3 are optionally cyclically linked to form a 5 or 6-membered heterocyclyl.
  • R 2 is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.
  • R 2 is hydrogen. In certain embodiments, R 2 is 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, R 2 is methyl. In certain embodiments, R 2 is 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, R 2 is alkynyl or substituted alkynyl.
  • R 2 is alkoxy or substituted alkoxy. In certain embodiments, R 2 is amino or substituted amino. In certain embodiments, R 2 is carboxyl or carboxyl ester. In certain embodiments, R 2 is acyl or acyloxy. In certain embodiments, R 2 is acyl amino or amino acyl. In certain embodiments, R 2 is alkylamide or substituted alkylamide. In certain embodiments, R 2 is sulfonyl. In certain embodiments, R 2 is thioalkoxy or substituted thioalkoxy.
  • R 2 is 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.
  • R 2 is 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.
  • R 2 is 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.
  • R 2 is 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.
  • R 3 is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.
  • R 3 is hydrogen. In certain embodiments, R 3 is 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, R 3 is methyl. In certain embodiments, R 3 is 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, R 3 is alkynyl or substituted alkynyl.
  • R 3 is alkoxy or substituted alkoxy. In certain embodiments, R 3 is amino or substituted amino. In certain embodiments, R 3 is carboxyl or carboxyl ester. In certain embodiments, R 3 is acyl or acyloxy. In certain embodiments, R 3 is acyl amino or amino acyl. In certain embodiments, R 3 is alkylamide or substituted alkylamide. In certain embodiments, R 3 is sulfonyl. In certain embodiments, R 3 is thioalkoxy or substituted thioalkoxy.
  • R 3 is 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.
  • R 3 is 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.
  • R 3 is 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.
  • R 3 is 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.
  • both R 2 and R 3 are methyl.
  • R 2 and R 3 are optionally cyclically linked to form a 5 or 6- membered heterocyclyl. In certain embodiments, R 2 and R 3 are cyclically linked to form a 5 or 6-membered heterocyclyl. In certain embodiments, R 2 and R 3 are cyclically linked to form a 5-membered heterocyclyl. In certain embodiments, R 2 and R 3 are cyclically linked to form a 6-membered heterocyclyl.
  • each R 4 is independently selected from hydrogen, halogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, amino, substituted amino, carboxyl, carboxyl ester, acyl, acyloxy, acyl amino, amino acyl, alkylamide, substituted alkylamide, sulfonyl, thioalkoxy, substituted thioalkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.
  • R 4 is hydrogen. In certain embodiments, each R 4 is hydrogen. In certain embodiments, R 4 is halogen, such as F, Cl, Br or I. In certain embodiments, R 4 is F. In certain embodiments, R 4 is Cl. In certain embodiments, R 4 is Br. In certain embodiments, R 4 is I. In certain embodiments, R 4 is alkyl or substituted alkyl, 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, R 4 is methyl.
  • R 4 is 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.
  • R 4 is alkynyl or substituted alkynyl.
  • R 4 is alkoxy or substituted alkoxy.
  • R 4 is amino or substituted amino.
  • R 4 is carboxyl or carboxyl ester.
  • R 4 is acyl or acyloxy.
  • R 4 is acyl amino or amino acyl.
  • R 4 is 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.
  • R 4 is 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.
  • R 4 is 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.
  • L A is a first linker. Examples of linkers that can be used in the conjugates of the present disclosure are described in more detail below.
  • L B is a second linker. Examples of linkers that can be used in the conjugates of the present disclosure are described in more detail below.
  • W 1 is 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.
  • W 2 is a second drug (or a second active agent).
  • 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.
  • Ab represents an antibody that binds to tissue factor (“TF antibody”).
  • TF antibody tissue factor
  • Ab comprises one or more fGly’ residues as described herein.
  • the TF antibody is attached to the rest of the conjugate through 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.
  • the conjugate of Formula (I) includes a first linker, L A .
  • the first linker, L A 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, L A may be bound (e.g., covalently bonded) to the conjugation moiety (e.g., as described herein).
  • the first linker, L A 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, L A , (and thus the first drug) to a TF antibody.
  • L A is attached to Ab through a conjugation moiety, and thus Ab is indirectly bonded to the linker L A through the hydrazinyl- indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety.
  • Ab is a TF antibody, and thus L A is attached through the hydrazinyl-indolyl or a hydrazinyl-pyrrolo- pyridinyl conjugation moiety to the TF antibody, e.g, the linker L A is indirectly bonded to the TF antibody through the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety.
  • the first linker L A may 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.
  • the first linker L A may include an alkyl or substituted alkyl group. In certain embodiments, the first linker L A may include an alkenyl or substituted alkenyl group. In certain embodiments, the first linker L A may include an alkynyl or substituted alkynyl group. In certain embodiments, the first linker L A may include an alkoxy or substituted alkoxy group. In certain embodiments, the first linker L A may include an amino or substituted amino group. In certain embodiments, the first linker L A may include a carboxyl or carboxyl ester group. In certain embodiments, the first linker L A may include an acyl amino group.
  • the first linker L A may include an alkylamide or substituted alkylamide group. In certain embodiments, the first linker L A may include an aryl or substituted aryl group. In certain embodiments, the first linker L A may include a heteroaryl or substituted heteroaryl group. In certain embodiments, the first linker L A may include a cycloalkyl or substituted cycloalkyl group. In certain embodiments, the first linker L A may include a heterocyclyl or substituted heterocyclyl group.
  • the first linker L A may include a polymer.
  • 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.
  • the polymer is a polyalkylene glycol.
  • the polymer is a polyethylene glycol.
  • Other linkers are also possible, as shown in the conjugates and compounds described in more detail below.
  • L A is a first linker described by the formula: -(L 1 )a-(L 2 )b-(L 3 )c-(L 4 )d-(L 5 ) e -(L 6 )f-, wherein L 1 , L 2 , L 3 , L 4 , L 5 and L 6 are each independently a linker subunit, and a, b, c, d, e and f are each independently 0 or 1, provided that at least one of a, b, c, d, e and f is 1. [00301] In certain embodiments, the sum of a, b, c, d, e and f is 1 to 6.
  • 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.
  • the linker subunit L 1 is attached to the hydrazinyl-indolyl or a hydrazinyl-pyrrolo-pyridinyl conjugation moiety (e.g., as shown in Formula (I) above).
  • the linker subunit L 2 if present, is attached to the first drug or active agent W 1 .
  • the linker subunit L 3 if present, is attached to the first drug or active agent W 1 .
  • the linker subunit L 4 if present, is attached to the first drug or active agent W 1 .
  • each of L 1 , L 2 , L 3 , L 4 , L 5 and L 6 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).
  • L 1 (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.
  • L 1 comprises a polyethylene glycol.
  • L 1 comprises a modified polyethylene glycol.
  • L 1 comprises an amino acid residue.
  • L 1 comprises an alkyl group or a substituted alkyl.
  • L 1 comprises an aryl group or a substituted aryl group.
  • L 1 comprises a diamine (e.g., a linking group comprising an alkylene diamine).
  • L 2 (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.
  • L 2 comprises a polyethylene glycol.
  • L 2 comprises a modified polyethylene glycol.
  • L 2 comprises an amino acid residue.
  • L 2 comprises an alkyl group or a substituted alkyl.
  • L 2 comprises an aryl group or a substituted aryl group.
  • L 2 comprises a diamine (e.g., a linking group comprising an alkylene diamine).
  • L 3 (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.
  • L 3 comprises a polyethylene glycol.
  • L 3 comprises a modified polyethylene glycol.
  • L 3 comprises an amino acid residue.
  • L 3 comprises an alkyl group or a substituted alkyl.
  • L 3 comprises an aryl group or a substituted aryl group.
  • L 3 comprises a diamine (e.g., a linking group comprising an alkylene diamine).
  • L 6 (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.
  • L 6 comprises a polyethylene glycol.
  • L 6 comprises a modified polyethylene glycol.
  • L 6 comprises an amino acid residue.
  • L 6 comprises an alkyl group or a substituted alkyl.
  • L 6 comprises an aryl group or a substituted aryl group.
  • L 6 comprises a diamine (e.g., a linking group comprising an alkylene diamine).
  • L 10 (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.
  • L 10 comprises a polyethylene glycol.
  • L 10 comprises a modified polyethylene glycol.
  • L 10 comprises an amino acid residue.
  • L 10 comprises an alkyl group or a substituted alkyl.
  • L 10 comprises an aryl group or a substituted aryl group.
  • L 10 comprises a diamine (e.g., a linking group comprising an alkylene diamine).
  • L B is a second linker comprising -(L 7 ) g -(L 8 )h-(L 9 )i-(L 10 )j-(L 11 )k-(L 12 )i-(L 13 )m-, where: -(L 7 ) g - is -(T 7 -V 7 ) g -;
  • 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.
  • 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.
  • the tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 and/or T 13 ) includes an aryl or substituted aryl.
  • the aryl can be phenyl.
  • the substituted aryl is a substituted phenyl.
  • (EDA) W includes 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.
  • each R 12 is independently selected from hydrogen, an alkyl, a substituted alkyl, an aryl and a substituted aryl.
  • any two adjacent R 12 groups of the EDA may be cyclically linked, e.g., to form a piperazinyl ring.
  • y is 1 and the two adjacent R 12 groups are an alkyl group, cyclically linked to form a piperazinyl ring.
  • y is 1 and the adjacent R 12 groups 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).
  • an alkyl e.g, methyl
  • a substituted alkyl e.g., lower alkyl-OH, such as ethyl-OH or propyl-OH.
  • the 4AP moiety is described by the structure: where R 12 is 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.
  • R 12 is a polyethylene glycol moiety.
  • R 12 is a carboxy modified polyethylene glycol.
  • a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 and/or T 13 ) includes (PEG)n, where (PEG)n is a polyethylene glycol or a modified polyethylene glycol linking unit.
  • (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.
  • n is 2.
  • n is 3.
  • n is 6.
  • n is 12.
  • 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.
  • 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.
  • p is 1.
  • p is 2.
  • R 13 is amino or substituted amino. In certain embodiments, R 13 is carboxyl or carboxyl ester. In certain embodiments, R 13 is acyl or acyloxy. In certain embodiments, R 13 is acyl amino or amino acyl. In certain embodiments, R 13 is alkylamide or substituted alkylamide. In certain embodiments, R 13 is sulfonyl. In certain embodiments, R 13 is thioalkoxy or substituted thioalkoxy.
  • R 13 is selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl.
  • alkyl, substituted alkyl, aryl, and substituted aryl are as described above for R 13 .
  • the tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 and/or T 13 ) includes an acetal group, a disulfide, a hydrazine, or an ester.
  • the tether group includes an acetal group.
  • the tether group includes a hydrazine.
  • the tether group includes a disulfide.
  • the tether group includes an ester.
  • a tether group (e.g., T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , T 12 and/or T 13 ) includes a 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), or para-hydroxy -phenyl (PHP).
  • MABO meta-amino-benzyloxy
  • MABC meta-amino- benzyloxycarbonyl
  • PABO para-amino-benzyloxycarbonyl
  • PABC para-amino-benzyloxycarbonyl
  • a tether group includes a PAB group described by the following structure:
  • a tether group includes a PAP group described by the following structure:
  • 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.
  • the glycoside or glycoside derivative is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.
  • 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.
  • the phenyl ring may be substituted with one or more additional groups selected from a glycoside and a glycoside derivative.
  • R 15 is 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.
  • R 15 is 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.
  • T 1 is selected from a (Ci-Ci2)alkyl and a substituted (Ci-Ci2)alkyl;
  • V 1 , V 2 , V 3 , V 4 ,V 5 and V 6 are each independently selected from a covalent bond, -CO-, -NR 15 -, -NR 15 (CH 2 )q-, -NR 15 (C 6 H 4 )-, -CONR 15 -, -NR 15 CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2-, -SO2NR 15 -, -NR 15 SO2- and -P(O)OH-, wherein q is an integer from 1 to 6; wherein: integer from 1 to 30;
  • L A comprises:
  • T 1 , T 2 , T 3 , T 4 , T 5 and T 6 are each independently selected from a covalent bond, (Ci- Cnjalkyl, substituted (Ci-Ci2)alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl, (EDA) W , (PEG)n, (AA) P , -(CR 13 OH) X -, 4-amino-piperidine (4AP), meta-amino-benzyloxy (MABO), meta-amino-benzyloxycarbonyl (MABC), para-amino-benzyloxy (PABO), para-amino- benzyloxycarbonyl (PABC), para-aminobenzyl (PAB), para-amino-benzylamino (PABA), para-amino-phenyl (PAP), para-hydroxy
  • T 1 is selected from a (Ci-Ci2)alkyl and a substituted (Ci-Ci2)alkyl;
  • T 2 , T 3 , T 4 , T 5 and T 6 are 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, -(CR 13 OH)X-, 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PAB A, PAP, PHP, an acetal group, a hydrazine, and an ester; and
  • V 1 , V 2 , V 3 , V 4 ,V 5 and V 6 are each independently selected from the group consisting of a covalent bond, -CO-, -NR 15 -, -NR 15 (CH 2 ) q -, -NR 15 (C 6 H 4 )-, -CONR 15 -, -NR 15 CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2- , -SO2NR 15 -, -NR 15 SO 2 -, and -P(O)OH-; wherein: integer from 1 to 30;
  • EDA is an ethylene diamine moiety having the following structure: integer from 1 to 6 and r is 0 or 1;
  • each R 12 is independently selected from hydrogen, an alkyl, a substituted alkyl, a polyethylene glycol moiety, an aryl and a substituted aryl, wherein any two adjacent R 12 groups may be cyclically linked to form a piperazinyl ring; a, b, c, and d are each 1; and e and f are 0.
  • T 1 , T 2 , T 3 , T 4 , T 5 and T 6 are each optionally substituted with a glycoside.
  • MABO, MABC, PABO, PABC, PAB, PABA, PAP and PHP are each optionally substituted with a glycoside.
  • the glycoside is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.
  • T 2 is (PEG)n and V 2 is -CO-;
  • T 3 is (AA) P and V 3 is absent;
  • T 1 is (Ci-Ci2)alkyl and V 1 is -CONH-;
  • T 3 is (AA) P and V 3 is absent;
  • T 4 is PABC and V 4 is absent; p is an integer from 1 to 10; and e and f are each 0; or wherein:
  • T 1 is (Ci-Ci2)alkyl and V 1 is -CONH-;
  • T 2 is (PEG)n and V 2 is -CO-;
  • T 5 is PABC and V 5 is absent; p is an integer from 1 to 10; and f is 0; or wherein:
  • T 3 is (Ci-Ci2)alkyl and V 3 is -O-;
  • T 4 is (Ci-Ci 2 )alkyl and V 4 is -CO-;
  • T 6 is PABC and V 6 is absent; or wherein:
  • T 2 is an amino acid analog and V 2 is absent;
  • T 3 is (AA) P and V 3 is absent;
  • T 1 is (Ci-Ci2)alkyl and V 1 is -CONH-;
  • T 2 is (PEG)n and V 2 is -CONH-;
  • T 1 is (Ci-Ci 2 )alkyl and V 1 is -CO-;
  • T 2 is an (AA) P and V 2 is -NH-;
  • T 3 is (PEG)n and V 3 is -CO-;
  • T 4 is (AA) P and V 4 is absent;
  • T 5 is PABC and V 5 is absent; p is an integer from 1 to 10; and f is 0; or wherein:
  • T 1 is (Ci-Ci2)alkyl and V 1 is -CONH-;
  • T 2 is (PEG)n and V 2 is -CO-;
  • T 4 is PAP and V 4 is -C(O)O-; p is an integer from 1 to 10; and e and f are each 0; or wherein:
  • AA is an amino acid residue, where p is an integer from 1 to 20; and each R 12 is independently selected from hydrogen, an alkyl, a substituted alkyl, a polyethylene glycol moiety, an aryl and a substituted aryl, wherein any two adjacent R 12 groups may be cyclically linked to form a piperazinyl ring; each R 13 is independently selected from hydrogen, alkyl, substituted alkyl, aryl, and substituted aryl; and each R 15 is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, carboxyl, carboxyl ester, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclyl, and substituted heterocyclyl.
  • 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.
  • the phenyl ring may be substituted with one or more additional groups selected from a glycoside and a glycoside derivative.
  • T 7 , T 8 , T 9 , T 10 , T 11 , T 12 and T 13 are each optionally substituted with a glycoside.
  • MABO, MABC, PABO, PABC, PAB, PABA, PAP and PHP are each optionally substituted with a glycoside.
  • the glycoside is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.
  • L B In some embodiments of L B : g, h, i, j, and k are each 1;
  • T 7 is a covalent bond
  • T 8 , T 9 , T 10 , T 11 and T 12 are 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, -(CR 13 OH)X-, 4-amino-piperidine (4AP), MABO, MABC, PABO, PABC, PAB, PABA, PAP, PHP, an acetal group, a hydrazine, and an ester; and
  • V 7 , V 8 , V 9 , V 10 ,V n and V 12 are each independently selected from the group consisting of a covalent bond, -CO-, -NR 15 -, -NR 15 (CH2)q-, -NR 15 (CeH4)-, -CONR 15 -, -NR 15 CO-, -C(O)O-, -OC(O)-, -O-, -S-, -S(O)-, -SO2- , -SO2NR 15 -, -NR 15 SO2-, and -P(O)OH-; wherein: integer from 1 to 30;
  • EDA is an ethylene diamine moiety having the following structure: integer from 1 to 6 and r is 0 or 1;
  • each R 12 is independently selected from hydrogen, an alkyl, a substituted alkyl, a polyethylene glycol moiety, an aryl and a substituted aryl, wherein any two adjacent R 12 groups may be cyclically linked to form a piperazinyl ring.
  • T 1 , T 2 , T 3 , T 4 , T 5 , T 6 , T 7 , T 8 , T 9 , T 10 , T 11 , and T 12 are each optionally substituted with a glycoside.
  • the glycoside is selected from a glucuronide, a galactoside, a glucoside, a mannoside, a fucoside, O-GlcNAc, and O-GalNAc.
  • T 7 , T 8 , T 9 , T 10 , T 11 , T 12 and T 13 and V 7 , V 8 , V 9 , V 10 ,V n , V 12 and V 13 are selected from the following: wherein:
  • T 10 is PABC and V 10 is absent; and k, 1 and m are each 0; or wherein:
  • T 8 is (Ci-Ci 2 )alkyl and V 8 is -CONH-;
  • T 11 is PABC and V 11 is absent;
  • T 12 is PABC and V 12 is absent; and m is 0; or wherein:
  • T 11 is PABC and V 11 is absent;
  • T 9 is substituted (Ci-Ci2)alkyl and V 9 is -CO-;
  • T 10 is (AA) P and V 10 is absent;
  • T 11 is PABC and V 11 is absent;
  • T 7 is absent and V 7 is -NHCO-;
  • T 8 is (Ci-Ci 2 )alkyl and V 8 is -CONH-;
  • T 9 is (PEG)n and V 9 is -CO-;
  • T 10 is (AA) P and V 10 is absent;
  • T 7 is absent and V 7 is -NHCO-;
  • T 8 is (Ci-Ci 2 )alkyl and V 8 is -CO-;
  • T 9 is 4AP and V 9 is -CO-;
  • T 10 is (Ci-Ci 2 )alkyl and V 10 is -CO-; T 11 is (AA) P and V 11 is absent;
  • T 12 is PABC and V 12 is absent; and m is 0; or wherein:
  • T 7 is absent and V 7 is -NHCO-;
  • T 8 is (Ci-Ci 2 )alkyl and V 8 is -CO-;
  • T 9 is 4AP and V 9 is -CO-;
  • T 10 is (Ci-Ci 2 )alkyl and V 10 is -O-;
  • T 11 is (Ci-Ci 2 )alkyl and V 11 is -CO-;
  • T 13 PABC and V 13 is absent; or wherein:
  • T 7 is absent and V 7 is -NHCO-;
  • T 9 is an amino acid analog and V 9 is absent;
  • T 8 is (Ci-Ci 2 )alkyl and V 8 is -CONH-;
  • T 9 is (PEG)n and V 9 is -CONH-;
  • T 10 is substituted (Ci-Ci2)alkyl and V 10 is -CO-;
  • T 11 is (AA) P and V 11 is absent;
  • T 12 is PABC and V 12 is absent; and m is 0; or wherein:
  • T 7 is absent and V 7 is -NHCO-;
  • T 8 is (Ci-Ci 2 )alkyl and V 8 is -CO-;
  • T 8 is (Ci-Ci 2 )alkyl and V 8 is -CONH-;
  • T 11 is PAP and V 11 is -C(O)O-;
  • T 10 is PABC and V 10 is absent; and k, 1 and m are each 0; or wherein:
  • T 7 is absent and V 7 is -NHCO-;
  • T 8 is (Ci-Ci2)alkyl and V 8 is absent;
  • T 11 is (PEG)n and V 11 is -CO-;
  • T 7 is absent and V 7 is -NHCO-;
  • T 9 is heteroaryl and V 9 is absent;
  • T 9 is heteroaryl and V 9 is absent;
  • T 10 is (Ci-Ci 2 )alkyl and V 10 is -C0NH-;
  • T 11 is substituted (Ci-Ci2)alkyl and V 11 is -CO-;
  • T 12 is (AA) P and V 12 is absent;
  • T 13 PABC and V 13 is absent.
  • the conjugate is an antibody-drug conjugate where the TF antibody and the drugs are linked together by linkers as described above.
  • the linker m(e.g., L A and/or L B ) is a cleavable linker.
  • a cleavable linker is a linker that includes one or more cleavable moieties, where the cleavable moiety includes one or more bonds that can dissociate under certain conditions, thus separating the cleavable linker into two or more separable portions.
  • 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.
  • 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.
  • 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.
  • 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.
  • a cleavable linker includes a first cleavable moiety and a second cleavable moiety that hinders cleavage of the first cleavable moiety.
  • 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 the cleavage of the cleavable linker.
  • the presence of uncleaved second cleavable moiety can hinder cleavage of the first cleavable moiety.
  • substantially means 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.
  • 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
  • the second cleavable moiety can protect the first cleavable moiety from cleavage.
  • 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.
  • 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 (z.e., cleavage of the first cleavable moiety is still needed in order to cleave the cleavable linker).
  • the cleavable moieties included in the cleavable linker may each be an enzymatically cleavable moiety.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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 (z.e., the site of the cancer (and overexpressed enzyme)).
  • 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.
  • the enzyme can be an esterase.
  • the enzymatically cleavable moiety is a cleavable moiety (e.g., an ester) that can be cleaved by an esterase enzyme.
  • the enzyme can be a glycosidase.
  • the enzymatically cleavable moiety is a cleavable moiety (e.g., a glycoside or glycoside derivative) that can be cleaved by a glycosidase enzyme.
  • the enzymatically cleavable moiety is an ester bond.
  • the first cleavable moiety described above i.e., the cleavable moiety protected from premature cleavage by the second cleavable moiety
  • the presence of uncleaved 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.
  • 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.
  • the second cleavable moiety includes a glycoside or glycoside derivative.
  • the enzymatically cleavable moiety is sugar moiety, such as a glycoside (or glyosyl) or glycoside derivative.
  • 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.
  • the second cleavable moiety (z.e., the cleavable moiety that protects the first cleavable moiety from premature cleavage) can be a glycoside or glycoside derivative.
  • the first cleavable moiety includes an ester and the second cleavable moiety includes a glycoside or glycoside derivative.
  • 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.
  • 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).
  • the glycosidic bond is an O-glycosidic bond (an O-glycoside).
  • 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 glycosidic bond that attaches the glycoside or glycoside derivative to the cleavable linker.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • a TF-ADC is represented by Formula (I): wherein:
  • L A is a first linker wherein:
  • the TF antibody comprises a heavy chain with a signal peptide and therefore have an amino acid sequence of:
  • 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.
  • 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
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 1, a VH CDR2 comprising the amino acid sequence of SEQ ID NO:2, a VH CDR3 comprising the amino acid sequence of SEQ ID NO:3, a VL CDR1 comprising the amino acid sequence of SEQ ID NO:4, a VL CDR2 comprising the amino acid sequence of SEQ ID NO:5, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO:6.
  • the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO:7, a VH CDR2 comprising the amino acid sequence of SEQ ID NO:2, a VH CDR3 comprising the amino acid sequence of SEQ ID NO:3, a VL CDR1 comprising the amino acid sequence of SEQ ID NO:4, a VL CDR2 comprising the amino acid sequence of SEQ ID NO:5, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO:6.
  • 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 NOV, 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 the amino acid sequence of SEQ ID NO: 12, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 13.
  • 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 NON, a VL CDR2 comprising the amino acid sequence of SEQ ID NO:5, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO:6.
  • 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.
  • 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.
  • 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.
  • the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NON 1, a VH CDR2 comprising the amino acid sequence of SEQ ID NON, a VH CDR3 comprising the amino acid sequence of SEQ ID NON, 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.
  • the TF antibody comprises a VH CDR1 comprising the amino acid sequence of SEQ ID NO:32, a VH CDR2 comprising the amino acid sequence of SEQ ID 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.
  • 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.
  • 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.
  • 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.
  • 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”).
  • IMGT® CDRs ImMunoGeneTics
  • 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 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.
  • 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.
  • 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 NON; (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.
  • 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 NON; (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.
  • 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:26.
  • 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.
  • 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:27, (ii) SEQ ID NO:31, (iii) SEQ ID NO:32, (iv) SEQ ID NO:35, and (v) SEQ ID NO:39; (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
  • 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:28, (ii) SEQ ID NO:33, (iii) SEQ ID NO:36, and (iv) SEQ ID NO:40; (2) a VL CDR2 comprising an amino acid sequence of any one of: (i) SEQ ID NO: 12, (ii) SEQ ID NO:29, and (iii) SEQ ID NO:37; and (3) a VL CDR3 comprising an amino acid sequence of any one of: (i) SEQ ID NO:30, (ii) SEQ ID NO:34, and (iii) SEQ ID NO:38.
  • a TF-ADC comprises a TF antibody, wherein the antibody comprises: a VL CDR1 comprising an amino acid sequence of any one of SEQ ID NOs:28, 33, 36, and 40; (2) a VL CDR2 comprising an amino acid sequence of any one of SEQ ID SEQ ID NOs: 12, 29, and 37; and (3) a VL CDR3 comprising an amino acid sequence of any one of SEQ ID SEQ ID NOs:30, 34, and 38.
  • 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:27; and (b) a VL region comprising: (1) a VL CDR1 comprising an amino acid sequence of any one of SEQ ID NOs:28, 33, 36, and 40; (2) a VL CDR2 comprising an amino acid sequence of any one of SEQ ID NOs: 12, 29, and 37; and (3) a VL CDR3 comprising an amino acid sequence of any one of SEQ ID SEQ ID NOs:30, 34, and 38.
  • 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:27; and (b) a VL region comprising: (1) a VL CDR1 comprising an amino acid sequence of SEQ ID NO:28; and (2) a VL CDR2 comprising an amino acid sequence of SEQ ID NO:29; and (3) a VL CDR3 comprising an amino acid sequence of SEQ ID SEQ ID NO:30.
  • a TF-ADC is prepared from a TF antibody, wherein the antibody comprises a heavy chain comprising an amino acid sequence of SEQ ID NO:41, and/or a light chain comprising an amino acid sequence of SEQ ID NO:42.
  • 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:42.
  • 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:92.
  • a TF-ADC comprises a TF antibody, wherein the antibody comprises a heavy chain that is conjugated to 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:42.
  • a TF-ADC comprises a TF antibody, wherein the antibody comprises a heavy chain that is conjugated to one or more fGly site(s) conjugated to a linker-drug 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:92.
  • the amino terminus of a VH and/or VL CDR1, CDR2, and/or CDR3 can be shortened by one, two, three, four, five or more amino acids compared to one or more of the CDRs described by SEQ ID NOs: 1-24 and 27-40, provided that binding to TF (e.g., human TF) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%).
  • TF e.g., human TF
  • the carboxy terminus of a VH and/or VL CDR1, CDR2, and/or CDR3 can be shortened by one, two, three, four, five or more amino acids compared to one or more of the CDRs described by SEQ ID NOs: 1-24 and 27-40, provided that binding to TF (e.g., human TF) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%). Any method known in the art can be used to ascertain whether binding to TF (e.g., human TF) is maintained, for example, the binding assays and conditions described in the “Examples” section described herein.
  • TF e.g., human TF
  • a TF-ADC comprises a TF antibody that comprises one or more (e.g., one, two, three, four, or more) conservative sequence modifications.
  • conservative sequence modifications include conservative amino acid substitutions in which an amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families are disclosed herein. Thus, in some embodiments, a predicted nonessential amino acid residue in a TF antibody can be replaced with another amino acid residue from the same side chain family.
  • the amino acid sequence modifications refer to at most one, two, three, four, five, or six amino acid substitutions to the CDRs, such as those described in any one of Tables 1-2.
  • each such CDR can contain up to five conservative amino acid substitutions, for example up to (not more than) four conservative amino acid substitutions, for example up to (not more than) three conservative amino acid substitutions, for example up to (not more than) two conservative amino acid substitutions, or no more than one conservative amino acid substitution.
  • the antibody in a TF-ADC comprises a VH comprising an amino acid sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:25 and a VL comprising an amino acid sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:26, and the binding of the antibody to TF (e.g., human TF) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%).
  • TF e.g., human TF
  • the antibody in a TF-ADC comprises a VH comprising an amino acid sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:41 and a VL comprising an amino acid sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:42, and the binding of the antibody to TF (e.g., human TF) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%).
  • TF e.g., human TF
  • the antibody in a TF-ADC comprises a heavy chain comprising an amino acid sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to any one of SEQ ID NOs:77-90 and a light chain comprising an amino acid sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:91 or 92, and the binding of the antibody to TF (e.g., human TF) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%).
  • TF e.g., human TF
  • the antibody in a TF-ADC comprises a heavy chain comprising an amino acid sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:93 and a light chain comprising an amino acid sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO:91, and the binding of the antibody to TF (e.g., human TF) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95%).
  • TF e.g., human TF
  • the antibody in the TF-ADC competes with the antibody designated as EXMA-006. Additionally or alternatively, the antibody in the TF-ADC competes with the antibody designated EXMA-007. In some embodiments, the antibody in the TF-ADC competes with any TF antibody as disclosed herein.
  • a cell can comprise one or more polynucleotides and/or one or more vectors.
  • a cell can be transformed or transfected with one or more polynucleotides encoding a TF antibody (e.g., a human TF antibody) or one or more vectors comprising the one or more polynucleotides encoding a TF antibody (e.g., a human TF antibody).
  • Methods for introducing DNA or RNA into a host cell include, but are not limited to, transformation, transfection, electroporation, nuclear injection, and fusion with carriers such as liposomes, micelles, ghost cells, and protoplasts.
  • host cells are useful for amplifying polynucleotides and for expressing polypeptides (e.g., antibodies) encoded by the polynucleotides.
  • a process to produce a TF antibody can comprise introducing RNA or DNA that encodes for a TF antibody, as described herein, into a host cell, culturing the host cell, and isolating the TF antibody thus produced.

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Abstract

La présente divulgation concerne des structures conjuguées anticorps-médicament d'anti-facteur tissulaire. Les structures conjuguées anticorps-médicament comprennent un lieur ramifié, au moins deux charges utiles par lieur ramifié étant fixées à un anticorps. La divulgation concerne également des composés et des méthodes de production de tels conjugués. La divulgation concerne, en outre, des méthodes d'utilisation des conjugués.
EP23863945.4A 2022-09-07 2023-09-06 Conjugués anticorps-médicament de facteur tissulaire et leurs utilisations Pending EP4583922A2 (fr)

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WO2025081117A2 (fr) * 2023-10-13 2025-04-17 Sutro Biopharma, Inc. Anticorps anti-facteur tissulaire et conjugués d'anticorps, compositions comprenant des anticorps anti-facteur tissulaire ou des conjugués d'anticorps, et méthodes de fabrication et d'utilisation d'anticorps anti-facteur tissulaire et de conjugués d'anticorps

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JP2025516569A (ja) * 2022-05-13 2025-05-30 エグゼリクシス， インコーポレイテッド ５ｔ４抗体薬物複合体及びその使用

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JP2025530129A (ja) 2025-09-11
KR20250060248A (ko) 2025-05-07
CO2025003047A2 (es) 2025-03-27
CL2025000595A1 (es) 2025-05-02
TW202423991A (zh) 2024-06-16
CA3265517A1 (fr) 2024-03-14
WO2024054821A3 (fr) 2024-04-11
AU2023338206A1 (en) 2025-03-06
PE20251256A1 (es) 2025-05-06
MX2025002614A (es) 2025-04-02
DOP2025000050A (es) 2025-04-08

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