Classifications

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  • 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
  • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
  • C07K16/2896—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against molecules with a "CD"-designation, not provided for elsewhere
• • 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
  • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
  • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
  • C07K16/2806—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD2
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
  • A61K35/48—Reproductive organs
  • A61K35/54—Ovaries; Ova; Ovules; Embryos; Foetal cells; Germ cells
  • A61K35/545—Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
• • 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
  • A61K47/6811—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a protein or peptide, e.g. transferrin or bleomycin
  • A61K47/6817—Toxins
• • 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
  • A61K47/6811—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a protein or peptide, e.g. transferrin or bleomycin
  • A61K47/6817—Toxins
  • A61K47/6831—Fungal toxins, e.g. alpha sarcine, mitogillin, zinniol or restrictocin
• • 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
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
  • A61P37/02—Immunomodulators
  • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
• • 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
  • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
  • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
  • C07K7/64—Cyclic peptides containing only normal peptide links
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
  • A61K2035/124—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells the cells being hematopoietic, bone marrow derived or blood cells
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
  • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
  • C07K2317/52—Constant or Fc region; Isotype
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
  • C07K2317/55—Fab or Fab'
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
  • C07K2317/71—Decreased effector function due to an Fc-modification
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
  • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
  • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
  • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
  • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
  • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
  • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

• the invention relates anti-CD1 17 antibodies, antibody drug conjugates (ADCs), and antigen-binding fragments thereof, as well as methods of treating patients suffering from various pathologies, such as blood diseases, metabolic disorders, cancers, and autoimmune diseases, among others, by administration of an antibody, or antibody drug conjugate (ADC), capable of binding an antigen expressed by a hematopoietic cell, such as a hematopoietic stem cell.
• ADC antibody drug conjugate
• hematopoietic stem cells have significant therapeutic potential, a limitation that has hindered their use in the clinic has been the difficulty associated with ensuring engraftment of hematopoietic stem cell transplants in a host.
• compositions that target specific endogenous stem cells that can be used as conditioning agents to promote the engraftment of exogenous hematopoietic stem cell grafts such that the multi-potency and hematopoietic functionality of these cells is preserved in the patient following transplantation.
• CD1 17 (also referred to as c-kit or Stem Cell Factor Receptor (SCRF)) is a single transmembrane, receptor tyrosine kinase that binds the ligand Stem Cell Factor (SCF) . SCF induces homodimerization of cKIT which activates its tyrosine kinase activity and signals through both the PI3-AKT and MAPK pathways (Kindblom et al., Am J. Path. 1998 152(5):1259).
• SCF Stem Cell Factor Receptor
• CD1 17 was initially discovered as an oncogene and has been studied in the field of oncology (see, for example, Stankov et al. (2014) Curr Pharm Des. 20(17):2849-80).
• An antibody drug conjugate (KTN0158) directed to CD1 17 is currently under investigation for the treatment of refractory gastrointestinal stromal tumors (GIST) (e.g., "KTN0158, a humanized anti-KIT monoclonal antibody, demonstrates biologic activity against both normal and malignant canine mast cells" London et al. (2016) Clin Cancer Res DOI: 10.1 158/1078-0432.CCR-16-2152).
• GIST refractory gastrointestinal stromal tumors
• CD1 17 is highly expressed on hematopoietic stem cells (HSCs). This expression pattern makes CD1 17 a potential target for conditioning across a broad range of diseases. There remains, however, a need for anti-CD1 17 based therapy that is effective for conditioning a patient for transplantation, such as a bone marrow transplantation.
• HSCs hematopoietic stem cells
• Described herein are antibodies, and antigen binding portions thereof, that specifically bind human CD1 17 (also known as c-kit), as well as compositions and methods of using said antibodies.
• the antibodies and fragments described herein can be used in anti-CD1 17 antibody drug conjugates (ADCs).
• the present invention provides compositions and methods for the direct treatment of various disorders of the hematopoietic system, metabolic disorders, cancers, and autoimmune diseases, among others.
• the invention additionally features methods for conditioning a patient, such as a human patient, prior to receiving hematopoietic stem cell transplant therapy so as to promote the engraftment of hematopoietic stem cell grafts.
• the patient may be one that is suffering from one or more blood disorders, such as a hemoglobinopathy or other hematopoietic pathology, and is thus in need of hematopoietic stem cell transplantation.
• hematopoietic stem cells are capable of differentiating into a multitude of cell types in the hematopoietic lineage, and can be administered to a patient in order to populate or re- populate a cell type that is deficient in the patient.
• the invention features methods of treating a patient with antibodies and antibody drug conjugates (ADCs) capable of binding proteins expressed by hematopoietic cells, such as CD1 17 (including, for example, GNNK+ CD1 17), so as to (i) directly treat a disease such as a blood disorder, metabolic disease, cancer, or autoimmune disease, among others described herein, by selectively depleting a population of cells that express CD1 17, such as an aberrant blood cell, cancer cell, or autoimmune cell, and/or (ii) deplete a population of endogenous hematopoietic stem cells within the patient.
• the former activity enables the direct treatment of a wide range of disorders associated with a cell of the hematopoietic lineage, as CD1 17 may be expressed by a cancerous cell, such as a leukemic cell, an
• autoimmune lymphocyte such as a T-cell that expresses a T-cell receptor that cross-reacts with a self antigen, among other cell types.
• the latter activity the selective depletion of hematopoietic stem cells, in turn creates a vacancy that can subsequently be filled by transplantation of an exogenous (for instance, an autologous, allogeneic, or syngeneic) hematopoietic stem cell graft.
• the invention thus provides methods of treating a variety of hematopoietic conditions, such as sickle cell anemia, thalassemia, Fanconi anemia, Wiskott-Aldrich syndrome, adenosine deaminase deficiency-severe combined immunodeficiency, metachromatic leukodystrophy, Diamond-Blackfan anemia and Schwachman-Diamond syndrome, human immunodeficiency virus infection, and acquired immune deficiency syndrome, as well as cancers and autoimmune diseases, among others.
• hematopoietic conditions such as sickle cell anemia, thalassemia, Fanconi anemia, Wiskott-Aldrich syndrome, adenosine deaminase deficiency-severe combined immunodeficiency, metachromatic leukodystrophy, Diamond-Blackfan anemia and Schwachman-Diamond syndrome, human immunodeficiency virus infection, and acquired immune deficiency syndrome, as well as cancers and autoimmune diseases
• the invention provides a method of depleting a population of CD1 17+ cells in a human patient by administering an effective amount of an antibody, antigen-binding fragment thereof, capable of binding CD1 17 where the antibody or fragment is conjugated to a cytotoxin (forming an ADC).
• the invention provides a method of depleting a population of CD1 17+ cells in a human patient in need of a hematopoietic stem cell transplant by administering, prior to the patient receiving a transplant including hematopoietic stem cells, an effective amount of an antibody or antigen-binding fragment thereof capable of binding CD1 17 conjugated to a cytotoxin (an ADC).
• the invention features a method, for example, of treating a human patient in need of a hematopoietic stem cell transplant, including administering to a human patient a transplant including hematopoietic stem cells, wherein the patient has been previously administered an antibody or antigen-binding fragment thereof capable of binding CD1 17 conjugated to a cytotoxin (forming an ADC) in an amount sufficient to deplete a population of CD1 17+ cells in the patient.
• an antibody or antigen-binding fragment thereof capable of binding CD1 17 conjugated to a cytotoxin (forming an ADC) in an amount sufficient to deplete a population of CD1 17+ cells in the patient.
• the invention features a method, for example, of treating a human patient in need of a hematopoietic stem cell transplant, including: administering to a human patient an antibody or antigen-binding fragment thereof capable of binding CD1 17 conjugated to a cytotoxin (forming an ADC) in an amount sufficient to deplete a population of CD1 17+ cells in the patient, and subsequently administering to the patient a transplant including hematopoietic stem cells.
• the cytotoxin may be, for example, pseudomonas exotoxin A, deBouganin, diphtheria toxin, an amatoxin, such as a-amanitin, saporin, maytansine, a
• auristatin an anthracycline, a calicheamicin, irinotecan, SN-38, a duocarmycin, a pyrrolobenzodiazepine, a pyrrolobenzodiazepine dimer, an indolinobenzodiazepine, or an indolinobenzodiazepine dimer, or a variant thereof.
• the cytotoxin is an amatoxin or derivative thereof, such as a-amanitin, ⁇ -amanitin, ⁇ -amanitin, ⁇ -amanitin, amanin, amaninamide, amanullin, amanullinic acid, and proamanullin.
• the cytotoxin is an amanitin.
• the cytotoxin is an amatoxin
• the antibody, or antigen-binding fragment thereof, conjugated to the cytotoxin is represented by the formula Ab-Z-L-Am, wherein Ab is the antibody, or antigen-binding fragment thereof, L is a linker, Z is a chemical moiety, and Am is the amatoxin.
• the amatoxin is conjugated to a linker.
• the amatoxin linker conjugate Am-L-Z is
• R 2 is H, OH, OR B , or OR c ;
• R A and R B when present, together with the oxygen atoms to which they are bound, combine to form an optionally substituted 5-membered heterocycloalkyl group;
• R 4 is H, OH, ORc, OR D , R c , or R D ;
• R 5 is H, OH, ORc, OR D , R c , or R D ;
• R 6 is H, OH, ORc, OR D , R c , or R D ;
• R 7 is H, OH, ORc, OR D , R c , or R D ;
• R 8 is OH, NH 2 , ORc, OR D , NHR C , or NR C R D ;
• R 9 is H, OH, ORc, or OR D ;
• X is -S-, -S(O)-, or -SO 2 -;
• Rc is -L-Z
• R D is optionally substituted alkyl (e.g., Ci-C 6 alkyl), optionally substituted heteroalkyl (e.g., Ci-C 6 heteroalkyl), optionally substituted alkenyl (e.g., C 2 -C 6 alkenyl), optionally substituted heteroalkenyl (e.g., C 2 -C 6 heteroalkenyl), optionally substituted alkynyl (e.g.
• C 2 -C 6 alkynyl optionally substituted heteroalkynyl (e.g., C 2 -C 6 heteroalkynyl), optionally substituted cycloalkyi, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl;
• Am contains exactly one R c substituent.
• linker L and the chemical moiety Z, taken together as L-Z is
• S is a sulfur atom which represents the reactive substituent present within an antibody,or antigen-binding fragment thereof, that binds CD1 17 (e.g., from the -SH group of a cysteine residue).
• Am-L-Z-Ab is one of:
• Am-L-Z-Ab is:
• R 2 is H, OH, OR B , or OR c ;
• R A and R B when present, together with the oxygen atoms to which they are bound, combine to form an optionally substituted 5-membered heterocycloalkyl group;
• R 4 is H, OH, ORc, OR D , R c , or R D ;
• R 5 is H, OH, ORc, OR D , R c , or R D ;
• R 6 is H, OH, ORc, OR D , R c , or R D ;
• R 7 is H, OH, ORc, OR D , R c , or R D ;
• R 8 is OH, NH 2 , ORc, OR D , NHR C , or NR C R D ;
• R 9 is H, OH, ORc, or OR D ;
• X is -S-, -S(O)-, or -SO 2 -;
• Rc is -L-Z
• R D is optionally substituted alkyl (e.g., C C 6 alkyl), optionally substituted heteroalkyl (e.g., C C 6 heteroalkyl), optionally substituted alkenyl (e.g., C 2 -C 6 alkenyl), optionally substituted heteroalkenyl (e.g., C 2 -C 6 heteroalkenyl), optionally substituted alkynyl (e.g., C 2 -C 6 alkynyl), optionally substituted heteroalkynyl (e.g., C 2 -C 6 heteroalkynyl), optionally substituted cycloalkyi, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl;
• alkyl e.g., C C 6 alkyl
• optionally substituted heteroalkyl e.g., C C 6 heteroalkyl
• optionally substituted alkenyl e.g., C 2 -C 6
• linker L and the chemical moiety Z, taken together as L-Z is
• R 2 is H, OH, OR B , or OR c ;
• R A and R B when present, together with the oxygen atoms to which they are bound, combine to form an optionally substituted 5-membered heterocycloalkyl group;
• R 3 is H, R c , or R D ;
• R 4 is H, OH, ORc, OR D , R c , or R D
• R 5 is H, OH, ORc, OR D , R c , or R D ;
• R 6 is H, OH, ORc, OR D , R c , or R D ;
• R 7 is H, OH, ORc, OR D , R c , or R D ;
• R 8 is OH, NH 2 , ORc, OR D , NHR C , or NR C R D ;
• R 9 is H, OH, ORc, or OR D ;
• X is -S-, -S(O)-, or -SO 2 -;
• Rc is -L-Z
• R D is optionally substituted alkyl (e.g., Ci-C 6 alkyl), optionally substituted heteroalkyl (e.g., Ci-C 6 heteroalkyl), optionally substituted alkenyl (e.g., C 2 -C 6 alkenyl), optionally substituted heteroalkenyl (e.g., C 2 -C 6 heteroalkenyl), optionally substituted alkynyl (e.g., C 2 -C 6 alkynyl), optionally substituted heteroalkynyl (e.g., C 2 -C 6 heteroalkynyl), optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl;
• alkyl e.g., Ci-C 6 alkyl
• heteroalkyl e.g., Ci-C 6 heteroalkyl
• optionally substituted alkenyl e.g., C 2 -C 6
• optionally substituted alkylene e.g., Ci-C 6 alkylene
• C C 6 heteroalkylene optional
• Z is a chemical moiety formed from a coupling reaction between a reactive substituent present on L and a reactive substituent present within an antibody, or antigen-binding fragment thereof, that binds CD1 17 (such as GNNK+ CD1 17);
• R A and R B together with the oxygen atoms to which they are bound, combine to form a 5-membered heterocyclo group of formula:
• R E and R E are each independently optionally substituted C C 6 alkylene-R c , optionally substituted C C 6 heteroalkylene-R c , optionally substituted C 2 -C 6 alkenylene-R c , optionally substituted C 2 -C 6 heteroalkenylene-R c , optionally substituted C 2 -C 6 alkynylene-R c , optionally substituted C 2 -C 6 heteroalkynylene-R c , optionally substituted cycloalkylene-R c , optionally substituted heterocycloalkylene-R c , optionally substituted arylene-R c , or optionally substituted heteroarylene-Rc.
• Am-L-Z is represented by formula (IA) or formula (IB), wherein R is H, OH, OR A , or OR c ;
• R 2 is H, OH, OR B , or OR c ;
• R A and R B together with the oxygen atoms to which they are bound, combine to form:
• R 3 is H or R c ;
• R 4 is H, OH, ORc, OR D , Rc, or R D
• R 5 is H, OH, ORc, OR D , Rc, or R D
• R 6 is H, OH, ORc, OR D , Rc, or R D
• R 7 is H, OH, ORc, ORD, Rc, or R D
• R 8 is OH, NH 2 , ORc, or NHR c ;
• R 9 is H or OH
• R c and R D are each as defined above.
• Am-L-Z is represented by formula (IA) or formula (IB),
• R is H, OH, OR A , or OR c ;
• R A and R B together with the oxygen atoms to which they are bound, combine to form:
• R 3 is H or R c ;
• R 4 and R 5 are each independently H, OH, OR c , Rc, or OR D ;
• R 6 and R 7 are each H
• R 8 is OH, NH 2 , ORc, or NHR C ;
• R 9 is H or OH
• R c is as defined above.
• Am-L-Z is represented by formula (IA) or formula (IB),
• R is H, OH, or OR A ;
• R 2 is H, OH, or OR B ;
• R A and R B together with the oxygen atoms to which they are bound, combine to form:
• R 3 , R 4 , R 6 , and R 7 are each H;
• R 5 is OR c ;
• R 8 is OH or NH 2 ;
• R 9 is H or OH;
• R c is as defined above.
• Am-L-Z is represented by formula (IA) or formula (IB),
• R and R 2 are each independently H or OH;
• R 3 is R c ;
• R 4 , R 6 , and R 7 are each H;
• R 5 is H, OH, or OC C 6 alkyl
• R 8 is OH or NH 2 ;
• R 9 is H or OH
• R c is as defined above.
• Am-L-Z is represented by formula (IA) or formula (IB),
• R 2 are each independently H or OH
• R 3 , R 6 , and R 7 are each H;
• R 4 and R 5 are each independently H, OH, OR c , or R c ;
• R 8 is OH or NH 2 ;
• R 9 is H or OH
• R c is as defined above.
• Am-L-Z is represented by formula (IA) or formula (IB),
• R and R 2 are each independently H or OH;
• R 3 , R 6 , and R 7 are each H;
• R 4 and R 5 are each independently H or OH
• R 8 is OH, NH 2 , ORc, or NHR C ;
• R 9 is H or OH
• R c is as defined above.
• linker L and the chemical moiety Z, taken together as L-Z is
• Ri is H or a linker covalently bound to the antibody or antigen-binding fragment thereof through a chemical moeity Z, formed from a coupling reaction between a reactive substituent present on the linker and a reactive substituent present within an antibody, or antigen- binding fragment thereof; and R 2 is H or a linker covalently bound to the antibody or antigen- binding fragment thereof through a chemical moeity Z, formed from a coupling reaction between a reactive substituent present on the linker and a reactive substituent present within an antibody, or antigen-binding fragment thereof; wherein when is H, R 2 is the linker, and when R 2 is H, is the linker.
• the linker comprises a -(CH) 2n - unit, where n is an integer from
• Ri is the linker and R 2 is H, and the linker and chemical moiety, together as L-Z, is
• Am-L-Z-Ab is:
• Am-L-Z-Ab is:
• the cytotoxin is a maytansinoid selected from the group consisting of DM1 and DM4.
• the cytotoxin is an auristatin selected from the group consisting of monomethyl auristatin E and monomethyl auristatin F.
• the cytotoxin is an anthracycline selected from the group consisting of daunorubicin, doxorubicin, epirubicin, and idarubicin.
• the invention features a method of depleting a population of CD1 17+ cells in a human patient by administering an effective amount of an antibody, an antigen-binding fragment, or an ADC thereof capable of binding GNNK+ CD1 17.
• the invention features a method of depleting a population of CD1 17+ cells in a human patient in need of a hematopoietic stem cell transplant by administering, prior to the patient receiving a transplant containing hematopoietic stem cells, an effective amount of an antibody or antigen-binding fragment thereof, or ADC capable of binding GNNK+ CD1 17.
• the invention features a method, for example, of treating a human patient in need of a hematopoietic stem cell transplant, including administering to a human patient a transplant containing hematopoietic stem cells, wherein the patient has been previously administered an antibody, antigen-binding fragment thereof, or ADC capable of binding GNNK+ CD1 17 in an amount sufficient to deplete a population of CD1 17+ cells in the patient.
• the invention features a method, for example, of treating a human patient in need of a hematopoietic stem cell transplant, including: administering to a human patient an antibody, antigen-binding fragment thereof, or ADC capable of binding GNNK+ CD1 17 in an amount sufficient to deplete a population of CD1 17+ cells in the patient, and subsequently administering to the patient a transplant including hematopoietic stem cells.
• the antibody, antigen-binding fragment thereof is selected from the group consisting of a monoclonal antibody or antigen-binding fragment thereof, a polyclonal antibody or antigen-binding fragment thereof, a humanized antibody or antigen-binding fragment thereof, a bispecific antibody or antigen-binding fragment thereof, a dual-variable immunoglobulin domain, a single-chain Fv molecule (scFv), a diabody, a triabody, a nanobody, an antibody-like protein scaffold, a Fv fragment, a Fab fragment, a F(ab') 2 molecule, and a tandem di-scFv.
• the antibody has an isotype selected from the group consisting of IgG, IgA, IgM, IgD, and IgE.
• the antibody, or antigen-binding fragment thereof is internalized by a hematopoietic cell, such as a hematopoietic stem cell, cancer cell, or autoimmune cell following administration to the patient.
• a hematopoietic cell such as a hematopoietic stem cell, cancer cell, or autoimmune cell following administration to the patient.
• the antibody, or antigen-binding fragment thereof, or ADC may be internalized by hematopoietic stem cells, cancer cells, or autoimmune cells by receptor-mediated endocytosis (e.g., upon binding to cell-surface CD1 17, such as GNNK+ CD1 17).
• a cytotoxin covalently bound to the antibody or antigen-binding fragment thereof may be released intracellularly by chemical cleavage (for instance, by enzymatic or non-specific cleavage of a linker described herein).
• the cytotoxin may then access its intracellular target (such as the mitotic spindle apparatus, nuclear DNA, ribosomal RNA, or topoisomerases, among others) so as to promote the death of an endogenous hematopoietic cell, such as an endogenous hematopoietic stem cell prior to transplantation therapy, an endogenous cancer cell, or an endogenous autoimmune cell, among others.
• the antibody, or antigen-binding fragment thereof, or ADC is capable of promoting necrosis of a hematopoietic cell, such as a hematopoietic stem cell, cancer cell, or autoimmune cell, among others.
• the antibody or antigen-binding fragment thereof may promote the death of an endogenous hematopoietic stem cell prior to transplantation therapy, an endogenous cancer cell, or an endogenous autoimmune cell, among others, by recruiting one or more complement proteins, natural killer (NK) cells, macrophages, neutrophils, and/or eosinophils to the cell, such as a hematopoietic stem cell upon administration to the patient.
• NK natural killer
• the transplant containing hematopoietic stem cells is administered to the patient after the concentration of the antibody, antigen-binding fragment thereof, or ADC has substantially cleared from the blood of the patient.
• the hematopoietic stem cells or progeny thereof maintain hematopoietic stem cell functional potential after two or more days (for example, from about 2 to about 5 days, from about 2 to about 7 days, from about 2 to about 20 days, from about 2 to about 30 days, such as 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 1 1 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, or more) following transplantation of the hematopoietic stem cells into the patient.
• days for example, from about 2 to about 5 days, from about 2 to about 7 days, from about 2 to about 20 days, from about 2 to about 30 days, such as 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 1 1 days, 12 days, 13
• the hematopoietic stem cells or progeny thereof are capable of localizing to hematopoietic tissue, such as the bone marrow, and/or reestablishing hematopoiesis following transplantation of the hematopoietic stem cells into the patient.
• the hematopoietic stem cells upon transplantation into the patient, give rise to recovery of a population of cells selected from the group consisting of megakaryocytes, thrombocytes, platelets, erythrocytes, mast cells, myeoblasts, basophils, neutrophils, eosinophils, microglia, granulocytes, monocytes, osteoclasts, antigen- presenting cells, macrophages, dendritic cells, natural killer cells, T-lymphocytes, and B- lymphocytes.
• a population of cells selected from the group consisting of megakaryocytes, thrombocytes, platelets, erythrocytes, mast cells, myeoblasts, basophils, neutrophils, eosinophils, microglia, granulocytes, monocytes, osteoclasts, antigen- presenting cells, macrophages, dendritic cells, natural killer cells, T-lymphocytes, and B- lymphocytes.
• the method is used to treat one or more disorders, such as by depleting a population of hematopoietic stem cells in a patient prior to hematopoietic stem cell transplant therapy so as to provide a niche to which the transplanted hematopoietic stem cells may home.
• the hematopoietic stem cells may establish productive hematopoiesis, so as to replenish a deficient cell type in the patient or a cell type that is being actively killed or has been killed, for instance, by chemotherapeutic methods.
• the patient may be one that is suffering from a stem cell disorder.
• the patient is suffering from a hemoglobinopathy disorder, such as sickle cell anemia, thalassemia, Fanconi anemia, aplastic anemia, and Wiskott-Aldrich syndrome.
• a hemoglobinopathy disorder such as sickle cell anemia, thalassemia, Fanconi anemia, aplastic anemia, and Wiskott-Aldrich syndrome.
• the patient may be suffering from an immunodeficiency disorder, such as a congenital
• immunodeficiency disorder or an acquired immunodeficiency disorder (e.g., human
• the patient is suffering from a metabolic disorder, such as glycogen storage diseases,
• the patient is suffering from a disorder selected from the group consisting of adenosine deaminase deficiency and severe combined immunodeficiency, hyper immunoglobulin M syndrome, Chediak-Higashi disease, hereditary lymphohistiocytosis, osteopetrosis, osteogenesis imperfecta, storage diseases, thalassemia major, systemic sclerosis, systemic lupus erythematosus, and juvenile rheumatoid arthritis.
• the patient is suffering from an autoimmune disease, such as scleroderma, multiple sclerosis, ulcerative colitis, Crohn's disease, ant Type 1 diabetes.
• the patient is suffering from cancer or myeloproliferative disease, such as a hematological cancer.
• the patient is suffering from acute myeloid leukemia (AML), acute lymphoid leukemia, chronic myeloid leukemia, chronic lymohoid leukemia, multiple meloma, diffuse large B- cell lymphoma, or non-Hodgkin's lymphoma.
• AML acute myeloid leukemia
• acute lymphoid leukemia chronic myeloid leukemia
• chronic lymohoid leukemia chronic lymohoid leukemia
• multiple meloma diffuse large B- cell lymphoma
• non-Hodgkin's lymphoma non-Hodgkin's lymphoma.
• the patient is suffering from a myelodysplastic disease, such as myelodysplastic syndrome.
• the method is used to directly treat a cancer, such as a cancer characterized by CD1 17+ cells (e.g., a leukemia characterized by CD1 17+ cells), by administration of an antibody, or antigen-binding fragment thereof, or ADC that depletes a population of CD1 17+ cancer cells in the patient and/or by administration of an antibody, or antigen-binding fragment thereof, so as to deplete a population of endogenous hematopoietic stem cells prior to hematopoietic stem cell transplantation.
• the transplantation may in turn re-constitute, for example, a population of cells depleted during the process of eradicating cancer cells.
• the cancer may be a hematological cancer, such as acute myeloid leukemia, acute lymphoid leukemia, chronic myeloid leukemia, chronic lymohoid leukemia, multiple meloma, diffuse large B-cell lymphoma, or non-Hodgkin's lymphoma.
• a hematological cancer such as acute myeloid leukemia, acute lymphoid leukemia, chronic myeloid leukemia, chronic lymohoid leukemia, multiple meloma, diffuse large B-cell lymphoma, or non-Hodgkin's lymphoma.
• the method is used to treat an autoimmune disease, such as by administration of an antibody, antigen-binding fragment thereof, or ADC so as to deplete a population of CD1 17+ autoimmune cells and/or by administration of an antibody, or antigen-binding fragment thereof, or ADC so as to deplete a population of
• transplantation may in turn re-constitute, for example, a population of cells depleted during the process of eradicating autoimmune cells.
• the autoimmune disease may be, for example, scleroderma, multiple sclerosis (MS), human systemic lupus (SLE), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), treating psoriasis, Type 1 diabetes mellitus (Type 1 diabetes), acute disseminated encephalomyelitis (ADEM), Addison's disease, alopecia universalis, ankylosing spondylitisis, antiphospholipid antibody syndrome (APS), aplastic anemia, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease (AIED), autoimmune lymphoproliferative syndrome (ALPS), autoimmune oophoritis, Balo disease, Behcet's disease, bullous pemphigoid, cardiomyopathy, Chagas' disease, chronic fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, Crohn's disease, cicatrical pemphigoid, coeliac
• the invention features a method of treating a hemoglobinopathy disorder, such as sickle cell anemia, thalassemia, Fanconi anemia, aplastic anemia, and Wiskott-Aldrich syndrome.
• a hemoglobinopathy disorder such as sickle cell anemia, thalassemia, Fanconi anemia, aplastic anemia, and Wiskott-Aldrich syndrome.
• an immunodeficiency disorder such as a congenital immunodeficiency disorder or an acquired immunodeficiency disorder (e.g., human immunodeficiency virus or acquired immune deficiency syndrome).
• the invention features a method of treating a metabolic disorder, such as glycogen storage diseases, mucopolysaccharidoses, Gaucher's Disease, Hurlers Disease, sphingolipidoses, and metachromatic leukodystrophy.
• a metabolic disorder such as glycogen storage diseases, mucopolysaccharidoses, Gaucher's Disease, Hurlers Disease, sphingolipidoses, and metachromatic leukodystrophy.
• the invention features a method of treating a disorder selected from the group consisting of adenosine deaminase deficiency and severe combined immunodeficiency, hyper immunoglobulin M syndrome, Chediak-Higashi disease, hereditary lymphohistiocytosis, osteopetrosis, osteogenesis imperfecta, storage diseases, thalassemia major, systemic sclerosis, systemic lupus erythematosus, and juvenile rheumatoid arthritis
• the invention features a method of treating an autoimmune disease, such as scleroderma, multiple sclerosis, ulcerative colitis, Chron's disease, ant Type 1 diabetes.
• the invention features a method of treating a cancer or myeloproliferative disease, such as a hematological cancer.
• the invention features a method of treating acute myeloid leukemia, acute lymphoid leukemia, chronic myeloid leukemia, chronic lymohoid leukemia, multiple meloma, diffuse large B-cell lymphoma, or non-Hodgkin's lymphoma.
• the patient is suffering from a myelodyplastic disease, such as myelodysplastic syndrome.
• the method may include the steps of administering an antibody, or antigen-binding fragment thereof, or ADC that binds CD1 17 (e.g., GNNK+ CD1 17) and/or a hematopoietic stem cell transplant according to the method of any of the above-described aspects and embodiments of the invention.
• CD1 17 e.g., GNNK+ CD1 17
• ADC that binds CD1 17 and/or a hematopoietic stem cell transplant according to the method of any of the above-described aspects and embodiments of the invention.
• the invention provides a method of treating cancer directly, such as a cancer characterized by CD1 17+ cells (e.g., a leukemia characterized by CD1 17+ cells).
• a cancer characterized by CD1 17+ cells e.g., a leukemia characterized by CD1 17+ cells.
• the method includes
• the cancer may be a hematological cancer, such as acute myeloid leukemia, acute lymphoid leukemia, chronic myeloid leukemia, chronic lymohoid leukemia, multiple meloma, diffuse large B-cell lymphoma, or non-Hodgkin's lymphoma.
• the invention provides a method of treating an autoimmune disease, such as multiple sclerosis (MS), human systemic lupus (SLE), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), treating psoriasis, Type
• an autoimmune disease such as multiple sclerosis (MS), human systemic lupus (SLE), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), treating psoriasis, Type
• Type 1 diabetes mellitus Type 1 diabetes
• ADAM acute disseminated encephalomyelitis
• ADAM additive disseminated encephalomyelitis
• APS antiphospholipid antibody syndrome
• APS antiphospholipid antibody syndrome
• aplastic anemia autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease (AIED), autoimmune lymphoproliferative syndrome (ALPS), autoimmune oophoritis, Balo disease, Behcet's disease, bullous pemphigoid, cardiomyopathy, Chagas' disease, chronic fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, Crohn's disease, cicatrical pemphigoid, coeliac sprue-dermatitis herpetiformis, cold agglutinin disease, CREST syndrome, Degos disease, discoid lupus, dysautonomia, endometriosis, essential mixed cryoglobulinemia, fibro
• the method includes administering an antibody, or antigen-binding fragment thereof, or ADC that binds CD1 17 (e.g., GNNK+ CD1 17).
• the invention features a method of depleting a population of CD1 17+ (e.g., GNNK+ CD1 17+) cells by contacting the population with an effective amount of a conjugate (or ADC) represented by the formula Ab-Z-L-Am, wherein Ab is an antibody or antigen-binding fragment thereof that binds CD1 17, Z is a chemical moiety, L is a linker and Am is an amatoxin.
• ADC conjugate
• Am-L-Z may be represented by formula (IA)
• R 2 is H, OH, OR B , or OR c ;
• R A and R B together with the oxygen atoms to which they are bound, combine to form an optionally substituted 5-membered heterocycloalkyl group;
• R 3 is H, R c , or R D ;
• R 4 , R 5 , R 6, and R 7 are each independently H, OH, OR c , OR D , R c , or R D ;
• R 8 is OH, NH 2 , ORc, OR D , NHR C , or NR C R D ;
• R 9 is H, OH, ORc, or OR D ;
• X is -S-, -S(O)-, or -SO 2 -;
• Rc is -L-Z
• R D is optionally substituted alkyl (e.g., Ci-C 6 alkyl), optionally substituted heteroalkyl (e.g., Ci-C 6 heteroalkyl), optionally substituted alkenyl (e.g., C 2 -C 6 alkenyl), optionally substituted heteroalkenyl (e.g., C 2 -C 6 heteroalkenyl), optionally substituted alkynyl (e.g., C 2 -C 6 alkynyl), optionally substituted heteroalkynyl (e.g., C 2 -C 6 heteroalkynyl), optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl;
• alkyl e.g., Ci-C 6 alkyl
• heteroalkyl e.g., Ci-C 6 heteroalkyl
• optionally substituted alkenyl e.g., C 2 -C 6
• Z is a chemical moiety formed from a coupling reaction between a reactive substituent present on L and a reactive substituent present within the antibody or antigen-binding fragment thereof,
• linker L and the chemical moiety Z, taken together as L-Z is
• R 2 is H, OH, OR B , or OR c ;
• R A and R B when present, together with the oxygen atoms to which they are bound, combine to form an optionally substituted 5-membered heterocycloalkyl group;
• R 3 is H, R c , or R D ;
• R 4 is H, OH, ORc, OR D , R c , or R D ;
• R 5 is H, OH, ORc, OR D , R c , or R D ;
• R 6 is H, OH, ORc, OR D , R c , or R D ;
• R 7 is H, OH, ORc, OR D , R c , or R D ;
• R 8 is OH, NH 2 , ORc, OR D , NHR C , or NR C R D ;
• R 9 is H, OH, ORc, or OR D ;
• X is -S-, -S(O)-, or -SO 2 -;
• Rc is -L-Z
• R D is optionally substituted alkyl (e.g., Ci-C 6 alkyl), optionally substituted heteroalkyl (e.g., Ci-C 6 heteroalkyl), optionally substituted alkenyl (e.g., C 2 -C 6 alkenyl), optionally substituted heteroalkenyl (e.g., C 2 -C 6 heteroalkenyl), optionally substituted alkynyl (e.g., C 2 -C 6 alkynyl), optionally substituted heteroalkynyl (e.g., C 2 -C 6 heteroalkynyl), optionally substituted cycloalkyi, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl;
• alkyl e.g., Ci-C 6 alkyl
• heteroalkyl e.g., Ci-C 6 heteroalkyl
• optionally substituted alkenyl e.g., C 2 -C 6
• optionally substituted alkylene e.g., C C 6 alkylene
• C C 6 heteroalkylene optionally substituted al
• Z is a chemical moiety formed from a coupling reaction between a reactive substituent present on L and a reactive substituent present within an antibody, or antigen-binding fragment thereof, that binds CD1 17 (such as GNNK+ CD1 17);
• linker L and the chemical moiety Z, taken together as L-Z is
• Am-L-Z-Ab is selected from
• Am-L-Z-Ab is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl
• the invention features a conjugate represented by the formula Ab-Z-L- Am, wherein Ab is an antibody or antigen-binding fragment thereof that binds CD1 17 (e.g., GNNK+ CD1 17) and Am is an amatoxin.
• Am-L-Z is represented by formula (I), (IA), (IB), (II), (HA), (IIB), (IV), (IVA), or (IVB), above.
• the antibody or antigen-binding fragment thereof is conjugated to the amatoxin by way of a cysteine residue in the Fc domain of the antibody or antigen-binding fragment thereof.
• the cysteine residue is introduced by way of a mutation in the Fc domain of the antibody or antigen-binding fragment thereof.
• the cysteine residue may be selected from the group consisting of Cys1 18, Cys239, and Cys265.
• the cysteine residue is naturally occurring in the Fc domain of the antibody or antigen-binding fragment thereof.
• the Fc domain may be an IgG Fc domain, such as a human lgG1 Fc domain, and the cysteine residue may be selected from the group consisting of Cys261 , Csy321 , Cys367, and Cys425.
• R is H, OH, or OR A ;
• R 2 is H, OH, or OR B ;
• R A and R B together with the oxygen atom which they are bound, combine to form:
• R 3 , R 4 , R 6 , and R 7 are each H;
• R 5 is OR c ;
• R 8 is OH or NH 2 ;
• R 9 is H or OH
• X is -S-, -S(O)-, or -S0 2 -.ln some embodiments, R and R 2 are each independently H or
• R 3 is R c ;
• R 4 , R 6 , and R 7 are each H;
• R 5 is H, OH, or OC C 6 alkyl
• R 8 is OH or NH 2 ;
• R 9 is H or OH
• Ri and R 2 are each independently H or
• R 3 , R 6 , and R 7 are each H;
• R 4 is ORc, or R c ;
• R 5 is H, OH, or OC C 6 alkyl
• R 8 is OH or NH 2 ;
• R 9 is H or OH
• X is -S-, -S(O)-, or -S0 2 -.ln some embodiments, R and R 2 are each independently H or
• R 3 , R 6 , and R 7 are each H;
• R 4 and R 5 are each independently H or OH
• R 8 is ORc or NHR C ;
• R 9 is H or OH
• X is -S-, -S(O)-, or -S0 2 -.ln some embodiments of these aspects, the antibody or antigen- binding fragment thereof is internalized by a CD1 17+ cell.
• the antibody or antigen-binding fragment thereof binds CD1 17 with a K d of less than 1 ⁇ , less than 750 nM, less than 500 nM, less than 250 nM, less than 200 nM, less than 150 nM, less than 00 nM, less than 75 nM, less than 50 nM, less than 10 nM, less than 1 nM, less than 0.1 nM, less than 10 pM, less than 1 pM, or less than 0.1 pM, In some embodiments, the K d is from about 0.1 pM to about 1 ⁇ .
• the antibody or antigen-binding fragment thereof binds CD1 17 with a k on of from about 9 x 10 "2 M “1 s “1 to about 1 x 10 2 M “1 s “1 .
• the antibody or antigen-binding fragment thereof competitively inhibits the binding of CD1 17 to a second antibody or antigen binding fragment thereof, or binds the same epitope as a second antibody, wherein the second antibody or antigen- binding fragment thereof has the following complementarity determining regions (CDRs):
• the disclosure further provides isolated anti-CD1 17 antibodies that may be used in the antibody drug conjugates (ADCs) disclosed herein comprising heavy and light chain CDRs and variable regions described in Table 1 , Table 6, or Table 8.
• ADCs antibody drug conjugates
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 8.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 9.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 9.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 9.
• CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 10.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 1 1 .
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 13.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 14.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 14.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 14.
• CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 15.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 16.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 18.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 19.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 19.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 19.
• CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 20.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 21 .
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 23.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 24, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 25.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 26, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 27.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 28, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 29.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 30, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 31 .
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO:
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 36, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 37.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 38, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 39.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 40, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 41 .
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 32, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 42.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO:
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 47, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 48.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 49, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 50.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 51 , and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 52.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 53, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 54.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO:
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 59, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 60.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 61 , and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 50.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 62, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 63.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 64, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 65.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO:
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 70, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 71 .
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 72, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 73.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 74, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 75.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 76, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 77.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO:
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 82, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 83.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 84, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 85.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 86, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 87. In one embodiment, the anti-CD1 17 antibody, or antigen binding portion thereof, comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 88. In one embodiment, the anti-CD1 17 antibody, or antigen binding portion thereof, comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 90.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 91 .
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 91 .
• CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 92.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 93.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO:95.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 96.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 96.
• CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 97.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 97.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO:
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 143, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 156.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 159, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 156.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 160, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 152.
• the anti- CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 98, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 99.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 99.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 100.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 98, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 101 .
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 98, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 102.
• the antibody, or antigen-binding fragment thereof is selected from the group consisting of a monoclonal antibody or antigen-binding fragment thereof, a polyclonal antibody or antigen-binding fragment thereof, a humanized antibody or antigen-binding fragment thereof, a bispecific antibody or antigen-binding fragment thereof, a dual- variable immunoglobulin domain, a single-chain Fv molecule (scFv), a diabody, a triabody, a nanobody, an antibody-like protein scaffold, a Fv fragment, a Fab fragment, a F(ab') 2 molecule, and a tandem di-scFV.
• the antibody is an intact antibody.
• the invention features a conjugate (ADC) represented by the formula Ab-Z-L-Cy, wherein Ab is an antibody or antigen-binding fragment thereof that binds CD1 17 (e.g., GNNK+ CD1 17) and Cy is a cytotoxin.
• the cytotoxin is pseudomonas exotoxin A, deBouganin, diphtheria toxin, saporin, maytansine, a maytansinoid, an auristatin, an anthracycline, a calicheamicin, irinotecan, SN-38, a duocarmycin, a
• pyrrolobenzodiazepine a pyrrolobenzodiazepine dimer, an indolinobenzodiazepine, or an indolinobenzodiazepine dimer, or a variant of any of the foregoing cytotoxins.
• the antibody, antigen-binding fragment thereof, or ADC is internalized by a CD1 17+ cell.
• the antibody, antigen-binding fragment thereof, or ADC binds CD1 17 with a K d of less than 1 ⁇ , less than 750 nM, less than 500 n , less than 250 nM, less than 200 nM, less than 150 nM, less than 100 nM, less than 75 nM, less than 50 nM, less than 10 nM, less than 1 nM, less than 0.1 nM, less than 10 pM, less than 1 pM, or less than 0.1 pM measured by bio-layer interferometry (BLi).
• the K d is from about 0.1 pM to about 1 ⁇ .
• the antibody, antigen-binding fragment thereof, or ADC binds CD1 17 with a k on of from about 9 x 10 "2 M “1 s “1 to about 1 x 10 2 M “1 s “1 measured by a bio-layer interferometry (BLI) assay.
• BBI bio-layer interferometry
• an anti-CD1 17 antibody, or antigen binding fragment thereof has a certain dissociation rate which is particularly advantageous when used as a part of a conjugate.
• an anti-CD1 17 antibody has, in certain embodiments, an off rate constant (Kdis) for human CD1 17 of 1 x 10 "2 to 1 x 10 "7 , 1 x 10 "3 to 1 x 10 "7 , 1 x 10 "4 to 1 x 10 "7 , 1 x 10 "5 to 1 x 10 "7 , or 1 x 10 "6 to 1 x 10 "7 s "1 , measured by bio-layer interferometry (BLI).
• Kdis off rate constant
• the antibody, antigen-binding fragment thereof, or ADC competitively inhibits the binding of CD1 17 to a second antibody or antigen binding fragment thereof, wherein the second antibody or antigen-binding fragment thereof has the following CDRs: a. a CDR-H1 having the amino acid sequence SYWIG (SEQ ID NO: 1 );
• a CDR-H2 having the amino acid sequence IIYPGDSDTRYSPSFQG (SEQ ID NO: 2);
• c. a CDR-H3 having the amino acid sequence HGRGYNGYEGAFDI (SEQ ID NO: 3);
• the antibody or antigen-binding fragment thereof is selected from the group consisting of a monoclonal antibody or antigen-binding fragment thereof, a polyclonal antibody or antigen-binding fragment thereof, a humanized antibody or antigen-binding fragment thereof, a bispecific antibody or antigen-binding fragment thereof, a dual-variable immunoglobulin domain, a single-chain Fv molecule (scFv), a diabody, a triabody, a nanobody, an antibody-like protein scaffold, a Fv fragment, a Fab fragment, a F(ab') 2 molecule, and a tandem di- scFv.
• the antibody has an isotype selected from the group consisting of IgG, IgA, IgM, IgD, and IgE.
• the foregoing methods and compositions include an isolated anti- CD1 17 antibody or antigen-binding fragment thereof comprising the CDRs set forth in the heavy and light chain amino acid sequences set forth in Table 1 , Table 6, or Table 8.
• the foregoing methods and compositions include an anti-CD1 17 antibody or antigen-binding fragment thereof comprising the variable regions set forth in the heavy and light chain amino acid sequences set forth in Table 1 .
• the foregoing methods and compositions include an lgG1 anti-CD1 17 antibody or antigen-binding fragment thereof comprises the variable regions set forth in the heavy and light chain amino acid sequences set forth in Table 1 , Table 6, or Table 8.
• the invention features a method treating acute myeloid leukemia (AML) in a human patient, the method comprising administering an effective amount of an anti-CD1 17
• AML acute myeloid leukemia
• the anti-CD1 17 ADC comprises an anti-CD1 17 antibody conjugated to a cytotoxin.
• the cytotoxin is an RNA polymerase inhibitor.
• the RNA polymerase inhibitor is an amatoxin.
• anti-CD1 17 antibody comprises the CDR sequences set forth in the heavy and light chain amino acid sequences of Table 1 .
• the anti-CD1 17 antibody is an intact antibody.
• the antibody is an lgG1 or an lgG4.
• the invention provides a conjugate represented by the formula Ab-Z-L- Am, wherein Ab is an antibody or antigen-binding fragment thereof that binds CD1 17, L is a linker, Z is a chemical moiety, and Am is an amatoxin, wherein the antibody or antigen-binding fragment thereof comprises the CDRs set forth in the heavy and light chain amino acid sequences of Table 1 , Table 6 or Table 8.
• linker- chemical moiety-amatoxin portion (Am-L-Z) of the conjugate is represented by formula
• R 2 is H, OH, OR B , or OR c ;
• R A and R B when present, together with the oxygen atoms to which they are bound, combine to form an optionally substituted 5-membered heterocycloalkyl group;
• R 3 is H, R c , or R D ;
• R 4 , R 5 , R 6, and R 7 are each independently H, OH, OR c , OR D , R c , or R D ;
• R 8 is OH, NH 2 , ORc, OR D , NHR C , or NR C R D ;
• R 9 is H, OH, ORc, or OR D ;
• X is -S-, -S(O)-, or -S0 2 -;
• Rc is -L-Z
• R D is optionally substituted C C 6 alkyl, optionally substituted C C 6 heteroalkyl, optionally substituted C 2 -C 6 alkenyl, optionally substituted C 2 -C 6 heteroalkenyl, optionally substituted C 2 -C 6 alkynyl, optionally substituted C 2 -C 6 heteroalkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl;
• Z is a chemical moiety formed from a coupling reaction between a reactive substituent present on L and a reactive substituent present within the antibody or antigen-binding fragment thereof,
• R 2 is H, OH, OR B , or OR c ;
• R A and R B when present, together with the oxygen atoms to which they are bound, combine to form an optionally substituted 5-membered heterocycloalkyl group;
• R 3 is H, R c , or R D ;
• R 4 , R 5 , R 6, and R 7 are each independently H, OH, OR c , OR D , R c , or R D ;
• R 8 is OH, NH 2 , ORc, OR D , NHR C , or NR C R D ;
• R 9 is H, OH, ORc, or OR D ;
• X is -S-, -S(O)-, or -S0 2 -;
• Rc is -L-Z
• R D is optionally substituted Ci-C 6 alkyl, optionally substituted Ci-C 6 heteroalkyl, optionally substituted C 2 -C 6 alkenyl, optionally substituted C 2 -C 6 heteroalkenyl, optionally substituted C 2 -C 6 alkynyl, optionally substituted C 2 -C 6 heteroalkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl;
• Z is a chemical moiety formed from a coupling reaction between a reactive substituent present on L and a reactive substituent present within the antibody or antigen-binding fragment thereof,
• linker- chemical moiety-amatoxin portion (Am-L-Z) of the conjugate is one of:
• linker- chemical moiety-amatoxin portion (Am-L-Z) of the conjugate is shown to indicate the point of attachment of the antibody.
• linker- chemical moiety-amatoxin portion (Am-L-Z) of the conjugate is
• linker- chemical moiety-amatoxin portion (Am-L-Z) of the conjugate is
• the Am-L-Z precursor is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-N-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-
• maleimide reacts with a thiol group found on a cysteine in the antibody.
• X is S, SO, or S0 2 ;
• Ri is H or a linker covalently bound to the antibody or antigen-binding fragment thereof through a chemical moeity Z, formed from a coupling reaction between a reactive substituent present on the linker and a reactive substituent present within an antibody, or antigen-binding fragment thereof;
• R 2 is H or a linker covalently bound to the antibody or antigen-binding fragment thereof through a chemical moeity Z, formed from a coupling reaction between a reactive substituent present on the linker and a reactive substituent present within an antibody, or antigen-binding fragment thereof;
• R 2 is the linker, and when R 2 is H, R is the linker.
• Ab-Z-L-Am is
• Ab-Z-L-Am is
• Fig. 1 demonstrates the measurement of binding by Bio-Layer Interferometry (BLI) of the indicated purified IgG (sensor-associated) to purified human CD1 17 ectodomain (R&D Systems #332-SR) at concentrations of 33.3 nM (top traces) and 1 1 nM (bottom traces) as a function of time.
• BLI Bio-Layer Interferometry
• the purified IgGs correspond to Ab1 (i.e., 001 ), Ab2 (i.e., 002), Ab3 (i.e., 003), Ab4 (i.e., 004), Ab5 (i.e., 005), Ab6 (i.e., 006), Ab7 (i.e., 007), Ab8 (i.e., 008), Ab9 (i.e., 009), Ab10 (i.e., 010), Ab1 1 (i.e., 01 1 ), Ab12 (i.e., 012), Ab13 (i.e., 013), Ab14 (i.e., 014), Ab15 (i.e., 015), and Ab16 (i.e., 016).
• Ab1 1 i.e., 01 1
• Ab12 i.e., 012
• Ab13 i.e., 013
• Ab14 i.e., 014
• Ab15 i.
• Figs. 2A and 2B graphically depict the results of in vitro cell killing assays that show Kasumi-1 cell viability as measured in luminescene (RLU) by Celltiter Glo (Fig. 2A) or viable CD34+ CD90+ cell count (Fig. 2B) in the presence of CD1 17-ADC or controls (y-axis) as a function of CD1 17-ADC or controls concentration (x-axis).
• Figs. 3A, 3B, 3C and 3D graphically depict the results of an in vivo cell depletion assay that shows CD1 17-ADC selectively depletes human HSCs in humanized NSG mice.
• FIG. 1 Schematic of in vivo mouse model.
• B Shows the percent of human myeloid or
• C the percentage of T cells present in the peripheral blood of CD1 17-ADC or control treated mice, expressed as a percent of that cell population prior to treatment (normalized to baseline).
• D Shows the absolute number of CD34+ cells in the bone marrow of CD1 17-ADC or control treated mice 21 days after a single administration of the ADC.
• Figs. 4A, 4B, 4C and 4D graphically depict the results of an in vivo tumor study that show CD1 17-ADC effectively depletes human leukemic cells in NSG mice.
• A Phenotypic analysis of Kasumi-1 cells in culture.
• B Phenotypic analysis of Kasumi-1 cells from bone marrow of tumor bearing mice at time of euthanasia.
• C Survival curve of mice treated 7 days after tumor injection with CD1 17-ADC or a naked anti-CD1 17 antibody or untreated controls.
• D Survival curve of mice treated 42 days after tumor injection with CD1 17-ADC or a naked anti-CD1 17 antibody or untreated controls.
• Figs. 5A, 5B and 5C graphically depict the results of an in vivo cell depletion assay that shows CD1 17-ADC selectively depletes HSCs in B6 mice.
• Fig. 6 graphically depicts the results of a non-human primate pharmacokinetic assay expressed as the concentration (ng/mL) of an isotype control antibody (i.e., "wild type antibody") in comparison to an CK6 variant antibody with a shorter half life as a function time (i.e., hours post- administration; x-axis).
• an isotype control antibody i.e., "wild type antibody”
• Figs. 7A and 7B graphically depict the results of an in vivo depletion assay showing that CD1 17-ADC effectively depletes human CD34+ cells in humanized NSG mice.
• A Phenotypic analysis of human cells in NSG bone marrow at day 21 when treated with a single dose of (i) wild type anti-CD1 17-ADC, (ii) a CK6 variant anti-CD1 17-ADC with a shorter half life or (iii) a control.
• FIG. B Shows the results of a humanized NSG mouse depletion assay after the administration of a single dose of a wild type anti-CD1 17-ADC (mg/kg) in comparison to a CK6 variant anti-CD1 17- ADC with a shorter half life (mg/kg) and various controls (i.e., "control,” “isotype-ADC (1 mg/kg)” and "Anti-CD1 17 (1 mg/kg)”), expressed as the CD34+ cell number per femur (y-axis) as a function of a single injection of (i) wild type anti-CD1 17-ADC, (ii) "short half life” variant anti- CD1 17-ADC or (iii) the controls (x-axis).
• Figs. 8A, 8B and 8C graphically depict the results of an in vivo depletion assay showing that human peripheral lymphocytes are maintained 21 days after administration of a single dose of anti-CD1 17 ADC in a humanized mouse model.
• A Shows the percentage of human CD33+ cells (relative to baseline) maintained after 21 days in mice treated with various concentrations of a single dose of (i) a wild type anti-CD1 17-ADC (mg/kg), (ii) a CK6 variant anti-CD1 17-ADC with a shorter half life (mg/kg) or (iii) various controls (i.e., "control,” “isotype-ADC (1 mg/kg)" and "Anti- CD1 17 (1 mg/kg)").
• (B) Shows the percentage of human CD3+ cells (relative to baseline) maintained after 21 days in mice treated with various concentrations of a single dose of (i) a wild type anti-CD1 17-ADC (mg/kg), (ii) a CK6 variant anti-CD1 17-ADC with a shorter half life (mg/kg) or (iii) various controls (i.e., "control,” “isotype-ADC (1 mg/kg)" and "Anti-CD1 17 (1 mg/kg)”).
• (C) Shows the percentage of human CD19+ cells (relative to baseline) maintained after 21 days in mice treated with various concentrations of a single dose of (i) a wild type anti-CD1 17-ADC (mg/kg), (ii) a CK6 variant anti-CD1 17-ADC with a shorter half life (mg/kg) or (iii) various controls (i.e., "control,” “isotype-ADC (1 mg/kg)” and "Anti-CD1 17 (1 mg/kg)”).
• Figs. 9A, 9B, 9C and 9D describe measurement of anti-CD1 17 antibody binding by Bio- Layer Interferometry (BLI) of the indicated purified IgG (sensor-associated) to purified human CD1 17 ectodomain (R&D Systems #332-SR) at concentrations of 33.3 nM (top trace) and 1 1 nM (bottom trace) as a function of time for the (A) HC-1 /LC-1 (Ab1 ) antibody, (B) the HC-77/LC-77 (Ab 77) antibody, (C) the HC-79/LC-79 (Ab79) antibody, and (D) the HC-81/LC-81 (Ab81 ) antibody.
• BLI Bio- Layer Interferometry
• Figs. 10A and 10B demonstrate the measurement of binding by Bio-Layer Interferometry (BLI) of the indicated purified IgG (sensor-associated) to purified human CD1 17 ectodomain (R&D Systems #332-SR) at concentrations of 33.3 nM (top traces) and 1 1 nM (bottom traces) as a function of time for the (A) HC-85/LC-85 antibody and the (B) HC-1/LC-1 antibody.
• BBI Bio-Layer Interferometry
• Figs. 11 A, 11 B, 11 C, and 11 D provide the variable heavy (VH) and variable light (VL) chain region of the amino acid sequences of CK6, Ab85 and Ab249.
• A Depicts the alignment of the variable heavy (VH) chain regions of CK6 (SEQ ID NO: 161 ) and Ab85 (SEQ ID NO: 143).
• B Depicts the alignment of the variable light (VL) chain regions of CK6 (SEQ ID NO: 162) and Ab85 (SEQ ID NO: 144).
• C Depicts the alignment of the variable heavy (VH) chain regions of CK6 (SEQ ID NO: 161 ) and Ab249 (SEQ ID NO: 98).
• D Depicts the alignment of the variable light (VL) chain regions of CK6 (SEQ ID NO: 162) and Ab249 (SEQ ID NO: 102).
• Figs. 12A, 12B and 12C depict the binding by bio-layer interferometry (BLI) of the indicated purified IgG (sensor-associated) to 1 1 nm (bottom trace) and 33 nM (top trace) purified human CD1 17 ectodomain as a function of time for the (A) CK6 antibody, (B) the HC-85/LC-85 (Ab85) antibody and the (C) HC-249/LC-249 (Ab 249) antibody.
• B bio-layer interferometry
• Figs. 13A and 13B illustrate the fraction of acidic variants present in the indicated antibody under the indicated incubation conditions (x-axis) for (A) Day 7 (25 e C and 50 e C) and (B) Day 15 (25 e C and 50 e C) compared to T 0 as determined by capillary electrophoresis.
• Figs. 14A, 14B, 14C, 14D and 14E depict chromatograms demonstrating the elution profile of (A) the CK6 antibody, (B) the HC-77 / LC-77 (Ab77) antibody, (C) the HC-79 / LC-79 (Ab79) antibody, (D) the HC-81 / LC-81 (Ab81 ) antibody, and (E) the HC-85 / LC-85 (Ab85) antibody,under the indication incubation conditions (see legend) after analysis by hydrophobic interaction chromatography (HIC).
• HIC hydrophobic interaction chromatography
• Fig. 15 graphically depicts the results of in vitro cell killing assays that show Kasumi-1 cell viability as measured in luminescene (RLU) by Celltiter Glo as a function of the indicated anti- CD1 17 ADC or control concentration.
• the IC50 (M) was determined by non-linear regression 4- parameter fit in Graphpad Prism.
• Fig. 16 graphically depicts the results of in vitro cell killing assays that show viable CD34+ CD90+ cell count as a function of the indicated anti-CD1 17 ADC or control concentration.
• the IC50 (M) was determined by non-linear regression 4-parameter fit in Graphpad Prism.
• Figs. 17A and 17B graphically depict the results of an in vivo cell depletion assay that shows a 0.3 mg/kg dose of the HC-85/LC-85 antibody (i.e., Ab85) ADC selectively depletes human HSCs in humanized NSG mice.
• ADC Shows the percent of human myeloid cells present in the peripheral blood of Ab85-ADC or control treated mice, expressed as a percent of that cell population prior to treatment (normalized to baseline) is shown for samples collected on day 0, 7, and 14.
• Fig. 18 graphically depicts the results of an in vitro toxicity assay using human bone marrow CD34+ cells for an anti-CD1 17 antibody (CK6) site specifically conjugated to various cytotoxic payloads.
• the results show that the anti-CD1 17 (CK6) antibody conjugated to amanitin resulted in a >90% depletion of human HSCs in humanized mice at 0.3 mg/kg.
• These results also show that the calicheamicin and amanitin ADCs demonstrate comparable depletion of HSCs
• Fig. 19 graphically depicts hematopoietic stem and progenitor cell number in bone marrow.
• CD1 17-AM amanitin conjugated anti-CD1 17
• Fig. 20 graphically depicts hematopoietic stem and progenitor cell number in bone marrow.
• the results show that a single dose or a fractionated dose of amanitin conjugated anti-CD1 17 (HC-85-LC 85) modified with a fast half-life Fc (H435A) eliminates bone marrow HSCs in cynomolgus monkeys.
• Male cynomolgus monkeys received a single i.v. dose (0.1 or 0.3 mg/kg) or a fractionated i.v. dose (0.3 mg/kg and 0.2 mg/kg Q3D) of anti-CD1 17 ADC.
• Bone marrow HSC counts were determined by flow cytometry 7 days post administration.
• FIGs. 21 A, 21 B, and 21 C graphically depict the results of a survival study using three AML- patient-derived xenografts developed from FLT-3+NPM1 + AML samples showing survival was significantly increased in mouse models administered 0.3 mg/kg anti-CD1 17-amanitin or 1 mg/kg anti-CD1 17-amanitin.
• A Survival curve of AML-PDX developed from FLT-3+NPM1 + AML sample J000106132 after single intravenous administration of ADCs (anti-CD1 17-AM, isotype-AM), an unconjugated anti-CD1 17 antibody or PBS (control).
• Fig. 22 demonstrates the measurement of binding by Bio-Layer Interferometry (BLI) of the indicated purified IgG (sensor-associated) to 100 nM purified human CD1 17 ectodomain (R&D Systems #332-SR) as a function of time.
• BLI Bio-Layer Interferometry
• the purified IgGs correspond to Ab17 (i.e., 017), Ab18 (i.e., 018), Ab19 (i.e., 019), Ab20 (i.e., 020), Ab21 (i.e., 021 ), Ab22 (i.e., 022), Ab23 (i.e., 023), Ab24 (i.e., 024), Ab25 (i.e., 025), Ab27 (i.e., 027), and Ab28 (i.e., 028).
• Fig. 23 demonstrates the measurement of binding by Bio-Layer Interferometry (BLI) of the indicated purified IgG (sensor-associated) to 100 nM purified rhesus CD1 17 ectodomain as a function of time.
• the purified IgGs correspond to Ab17 (i.e., 017), Ab18 (i.e., 018), Ab19 (i.e., 019), Ab20 (i.e., 020), Ab21 (i.e., 021 ), Ab22 (i.e., 022), Ab23 (i.e., 023), Ab24 (i.e., 024), Ab25 (i.e., 025), Ab27 (i.e., 027), and Ab28 (i.e., 028).
• BBI Bio-Layer Interferometry
• Described herein are isolated anti-CD1 17 human antibodies that bind to human CD1 17.
• antibodies provided herein have many characteristics making them advantageous for therapy, including methods of conditioning human patients for stem cell transplantation.
• antibodies disclosed herein in certain embodiments, have high affinity and a law off rate for huamn CD1 17, as well as the ability to internalize in cells expressing CD1 17.
• certain of the antibodies presented herein have improved biophysical stability. These features also make the anti-CD1 17 antibodies disclosed herein advantageous for use in conjugates (antibody drug conjugates (ADCs)) for delivering cytotoxins to CD1 17 expressing cells.
• ADCs antibody drug conjugates
• the invention provides anti-CD1 17 antibodies, specifically isolated human anti-CD1 17 antibodies that bind to the ectodomain of human CD1 17.
• the binding regions of the isolated anti- CD1 17 antibodies identified herein are described below and in Table 1 , Table 6, and Table 8.
• the anti-CD1 17 antibodies and ADCs described herein can be used in methods of treating a variety of disorders, such as diseases of a cell type in the hematopoietic lineage, cancers, autoimmune diseases, metabolic disorders, and stem cell disorders, among others.
• compositions and methods described herein may (i) directly deplete a population of cells that give rise to a pathology, such as a population of cancer cells (e.g., leukemia cells) and autoimmune cells (e.g., autoreactive T-cells), and/or (ii) deplete a population of endogenous hematopoietic stem cells so as to promote the engraftment of transplanted hematopoietic stem cells by providing a niche to which the transplanted cells may home.
• a pathology such as a population of cancer cells (e.g., leukemia cells) and autoimmune cells (e.g., autoreactive T-cells)
• autoimmune cells e.g., autoreactive T-cells
• deplete a population of endogenous hematopoietic stem cells so as to promote the engraftment of transplanted hematopoietic stem cells by providing a niche to which the transplanted cells may home.
• the foregoing activities
• this administration can cause a reduction in the quantity of the cells that give rise to the pathology of interest.
• this administration can cause the selective depletion of a population of endogenous hematopoietic stem cells, thereby creating a vacancy in the hematopoietic tissue, such as the bone marrow, that can subsequently be filled by transplanted, exogenous hematopoietic stem cells.
• the invention is based in part on the discovery that ADCs, antibodies, or antigen-binding fragments thereof, capable of binding CD1 17 (such as GNNK+ D1 17) can be administered to a patient to affect both of the above activities.
• ADCs, antibodies, or antigen-binding fragments thereof, that bind CD1 17 can be administered to a patient suffering from a cancer or autoimmune disease to directly deplete a population of cancerous cells or autoimmune cells, and can also be administered to a patient in need of hematopoietic stem cell transplant therapy in order to promote the survival and engraftment potential of transplanted hematopoietic stem cells.
• ADCs, antibodies, or antigen-binding fragments thereof can manifest in a variety of empirical measurements. For instance, engraftment of transplanted hematopoietic stem cells can be evaluated by assessing the quantity of competitive repopulating units (CRU) present within the bone marrow of a patient following administration of an ADC, antibody or antigen-binding fragment thereof capable of binding CD1 17 and subsequent administration of a hematopoietic stem cell transplant. Additionally, one can observe engraftment of a hematopoietic stem cell transplant by incorporating a reporter gene, such as an enzyme that catalyzes a chemical reaction yielding a fluorescent, chromophoric, or luminescent product, into a vector with which the donor
• a reporter gene such as an enzyme that catalyzes a chemical reaction yielding a fluorescent, chromophoric, or luminescent product
• hematopoietic stem cells have been transfected and subsequently monitoring the corresponding signal in a tissue into which the hematopoietic stem cells have homed, such as the bone marrow.
• a tissue into which the hematopoietic stem cells have homed such as the bone marrow.
• Engraftment can also be determined by measuring white blood cell counts in peripheral blood during a post-transplant period, and/or by measuring recovery of marrow cells by donor cells in a bone marrow aspirate sample.
• ADCs antibodies, or antigen-binding fragments thereof, that can be administered to a patient, such as a patient suffering from a cancer (such as acute myelogenous leukemia or myelodysplastic syndrome) or autoimmune disease, or a patient in need of hematopoietic stem cell transplant therapy in order to promote engraftment of hematopoietic stem cell grafts, as well as methods of administering such therapeutics to a patient (e.g., prior to hematopoietic stem cell transplantation).
• a cancer such as acute myelogenous leukemia or myelodysplastic syndrome
• autoimmune disease a patient in need of hematopoietic stem cell transplant therapy in order to promote engraftment of hematopoietic stem cell grafts, as well as methods of administering such therapeutics to a patient (e.g., prior to hematopoietic stem cell transplantation).
• the term “about” refers to a value that is within 10% above or below the value being described.
• the term “about 5 nM” indicates a range of from 4.5 nM to 5.5 nM.
• amatoxin refers to a member of the amatoxin family of peptides produced by Amanita phalloides mushrooms, or a variant or derivative thereof, such as a variant or derivative thereof capable of inhibiting RNA polymerase II activity.
• Amatoxins useful in conjunction with the compositions and methods described herein include compounds according to, but are not limited to, formula (III), including a-amanitin, ⁇ -amanitin, ⁇ -amanitin, ⁇ -amanitin, amanin, amaninamide, amanullin, amanullinic acid, or proamanullin.
• amatoxins may be conjugated to an antibody, or antigen-binding fragment thereof, for instance, by way of a linker moiety (L) (thus forming an ADC).
• L linker moiety
• Exemplary methods of amatoxin conjugation and linkers useful for such processes are described below.
• Exemplary linker-containing amatoxins useful for conjugation to an antibody, or antigen-binding fragment, in accordance with the compositions and methods are also described herein.
• R 2 is H, OH, or OR B ;
• R A and R B when present, together with the oxygen atoms to which they are bound, combine to form an optionally substituted 5-membered heterocycloalkyl group;
• R 3 is H or R D ;
• R 4 is H, OH, OR D , or R D ;
• R 5 is H, OH, OR D , or R D ;
• R 6 is H, OH, OR D , or R D ;
• R 7 is H, OH, OR D , or R D ;
• R 8 is OH, NH 2 , or OR D ;
• R 9 is H, OH, or OR D ;
• X is -S-, -S(O)-, or -SO 2 -;
• R D is optionally substituted alkyl (e.g., Ci-C 6 alkyl), optionally substituted heteroalkyi (e.g., Ci-C 6 heteroalkyi), optionally substituted alkenyl (e.g., C 2 -C 6 alkenyl), optionally substituted heteroalkenyl (e.g., C 2 -C 6 heteroalkenyl), optionally substituted alkynyl (e.g., C 2 -C 6 alkynyl), optionally substituted heteroalkynyl (e.g., C 2 -C 6 heteroalkynyl), optionally substituted cycloalkyi, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl.
• alkyl e.g., Ci-C 6 alkyl
• optionally substituted heteroalkyi e.g., Ci-C 6 heteroalkyi
• optionally substituted alkenyl e.g., C 2
• amatoxins useful in conjunction with the compositions and methods described herein include compounds according to formula (III A), below:
• R 2 is H, OH, or OR B ;
• R A and R B when present, together with the oxygen atoms to which they are bound, combine to form an optionally substituted 5-membered heterocycloalkyl group;
• R 3 is H or R D ;
• R 4 is H, OH, OR D , or R D
• R 5 is H, OH, OR D , or R D
• R 6 is H, OH, OR D , or R D
• R 7 is H, OH, OR D , or R D
• R 8 is OH, NH 2 , or OR D ;
• R 9 is H, OH, or OR D ;
• X is -S-, -S(O)-, or -SO 2 -;
• R D is optionally substituted alkyl (e.g., Ci-C 6 alkyl), optionally substituted heteroalkyi (e.g., C C 6 heteroalkyi), optionally substituted alkenyl (e.g., C 2 -C 6 alkenyl), optionally substituted heteroalkenyl (e.g., C 2 -C 6 heteroalkenyl), optionally substituted alkynyl (e.g., C 2 -C 6 alkynyl), optionally substituted heteroalkynyl (e.g., C 2 -C 6 heteroalkynyl), optionally substituted cycloalkyi, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl.
• alkyl e.g., Ci-C 6 alkyl
• optionally substituted heteroalkyi e.g., C C C 6 heteroalkyi
• optionally substituted alkenyl e.g., C 2
• NRB amino acids useful in conjunction with the compositions and methods described herein also include compounds according to formula (NIB), below:
• R 2 is H, OH, or OR B ;
• R A and R B when present, together with the oxygen atoms to which they are bound, combine to form an optionally substituted 5-membered heterocycloalkyl group;
• R 3 is H or R D ;
• R 4 is H, OH, OR D , or R D ;
• R 5 is H, OH, OR D , or R D ;
• R 6 is H, OH, OR D , or R D ;
• R 7 is H, OH, OR D , or R D ;
• R 8 is OH, NH 2 , or OR D ;
• R 9 is H, OH, or OR D ;
• X is -S-, -S(O)-, or -SO 2 -;
• R D is optionally substituted alkyl (e.g., Ci-C 6 alkyl), optionally substituted heteroalkyl (e.g., Ci-C 6 heteroalkyl), optionally substituted alkenyl (e.g., C 2 -C 6 alkenyl), optionally substituted heteroalkenyl (e.g., C 2 -C 6 heteroalkenyl), optionally substituted alkynyl (e.g., C 2 -C 6 alkynyl), optionally substituted heteroalkynyl (e.g., C 2 -C 6 heteroalkynyl), optionally substituted cycloalkyi, optionally substituted heterocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl.
• alkyl e.g., Ci-C 6 alkyl
• heteroalkyl e.g., Ci-C 6 heteroalkyl
• optionally substituted alkenyl e.g., C 2 -C 6
• amatoxins may be conjugated to an antibody, or antigen-binding fragment thereof, for instance, by way of a linker moiety (L) (thus forming an ADC).
• L linker moiety
• Exemplary methods of amatoxin conjugation and linkers useful for such processes are described below, including Table 3.
• Exemplary linker-containing amatoxins useful for conjugation to an antibody, or antigen-binding fragment, in accordance with the compositions and methods described herein are shown in structural formulas (I), (IA), (IB), (II), (HA), or (MB), recited herein.
• antibody refers to an immunoglobulin molecule that specifically binds to, or is immunologically reactive with, a particular antigen, and includes monoclonal, genetically engineered, and otherwise modified forms of antibodies, including but not limited to chimeric antibodies, humanized antibodies, heteroconjugate antibodies (e.g., bi- tri- and quad- specific antibodies, diabodies, triabodies, and tetrabodies), and antigen binding fragments of antibodies, including, for example, Fab', F(ab') 2 , Fab, Fv, rlgG, and scFv fragments.
• the antibodies of the present invention are generally isolated or recombinant.
• isolated when used herein refers to a polypeptide, e.g., an antibody, that has been identified and separated and/or recovered from a cell or cell culture from which it was expressed. Ordinarily, an isolated antibody will be prepared by at least one purification step. Thus, an “isolated antibody,” refers to an antibody which is substantially free of other antibodies having different antigenic specificities. For instance, an isolated antibody that specifically binds to CD1 17 is substantially free of antibodies that specifically bind antigens other than CD1 17.
• mAb monoclonal antibody
• mAb monoclonal antibody
• Fab and F(ab') 2 fragments refer to antibody fragments that lack the Fc fragment of an intact antibody. Examples of these antibody fragments are described herein.
• antibodies comprise heavy and light chains containing antigen binding regions.
• Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region.
• the heavy chain constant region is comprised of three domains, CH1 , CH2 and CH3.
• Each light chain is comprised of a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region.
• the light chain constant region is comprised of one domain, CL.
• the VH, and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR).
• CDR complementarity determining regions
• Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxyl-terminus in the following order: FR1 , CDR1 , FR2, CDR2, FR3, CDR3, FR4.
• the variable regions of the heavy and light chains contain a binding domain that interacts with an antigen.
• the constant regions of the antibodies can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system.
• an “intact” or “full length” antibody refers to an antibody having two heavy
• Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as HCVR or
• the heavy chain constant region is comprised of three domains, CH1 , CH2 and CH3. Each light chain is comprised of a light chain variable region
• LCVR LCVR
• VL light chain constant region
• the light chain constant region is comprised of one domain, CL.
• CL complementarity determining regions
• FR framework regions
• Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxyl-terminus in the following order: FR1 , CDR1 , FR2, CDR2, FR3, CDR3, FR4.
• the variable regions of the heavy and light chains contain a binding domain that interacts with an antigen.
• the constant regions of the antibodies can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system.
• Fc refers to the portion of an IgG antibody that correlates to a crystallizable fragment obtained by papain digestion of an IgG molecule.
• the Fc region comprises the C-terminal half of two heavy chains of an IgG molecule that are linked by disulfide bonds. It has no antigen binding activity but contains the carbohydrate moiety and binding sites for complement and Fc receptors, including the FcRn receptor.
• An Fc region contains the second constant domain CH2 (e.g., residues at EU positions 231 -340 of lgG1 ) and the third constant domain CH3 (e.g., residues at EU positions 341 -447 of human lgG1 ).
• the Fc region includes the "lower hinge region” (e.g., residues at EU positions 233- 239 of lgG1 ).
• Fc can refer to this region in isolation, or this region in the context of an antibody, antibody fragment, or Fc fusion protein. Polymorphisms have been observed at a number of positions in Fc domains, including but not limited to EU positions 270, 272, 312, 315, 356, and 358, and thus slight differences between the sequences presented in the instant application and sequences known in the art can exist.
• a "wild type IgG Fc domain” or "WT IgG Fc domain” refers to any naturally occurring IgG Fc region (i.e., any allele).
• sequences of the heavy chains of human lgG1 , lgG2, lgG3 and lgG4 can be found in a number of sequence databases, for example, at the Uniprot database (www.uniprot.org) under accession numbers P01857
• modified Fc region or “variant Fc region” as used herein refers to an IgG Fc domain comprising one or more amino acid substitutions, deletions, insertions or modifications introduced at any position within the Fc region.
• antigen-binding fragment refers to one or more fragments of an antibody that retain the ability to specifically bind to a target antigen.
• the antigen-binding function of an antibody can be performed by fragments of a full-length antibody.
• the antibody fragments can be, for example, a Fab, F(ab') 2 , scFv, diabody, a triabody, an affibody, a nanobody, an aptamer, or a domain antibody. Examples of binding fragments encompassed of the term
• antigen-binding fragment of an antibody include, but are not limited to: (i) a Fab fragment, a monovalent fragment consisting of the V L , V H , C L , and C H 1 domains; (ii) a F(ab') 2 fragment, a bivalent fragment containing two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the V H and C H 1 domains; (iv) a Fv fragment consisting of the V L and V H domains of a single arm of an antibody, (v) a dAb including V H and V L domains; (vi) a dAb fragment that consists of a V H domain (see, e.g., Ward et al., Nature 341 :544-546, 1989); (vii) a dAb which consists of a V H or a V L domain; (viii) an isolated complementarity determining region (CDR); and (ix)
• the two domains of the Fv fragment, V L and V H are coded for by separate genes, they can be joined, using recombinant methods, by a linker that enables them to be made as a single protein chain in which the V
• scFv single chain Fv
• These antibody fragments can be obtained using conventional techniques known to those of skill in the art, and the fragments can be screened for utility in the same manner as intact antibodies.
• Antigen-binding fragments can be produced by recombinant DNA
• anti-CD1 17 antibody or “an antibody that binds to CD1 17” refers to an antibody that is capable of binding CD1 17 with sufficient affinity such that the antibody is useful as a diagnostic and/or therapeutic agent in targeting CD1 17.
• bispecific antibody refers to, for example, a monoclonal, often a human or humanized antibody that is capable of binding at least two different antigens.
• hematopoietic stem cell surface antigen or another cell surface protein such as a receptor or receptor subunit involved in a signal transduction pathway that potentiates cell growth, among others.
• CDR complementarity determining region
• FRs framework regions
• the amino acid positions that delineate a hypervariable region of an antibody can vary, depending on the context and the various definitions known in the art. Some positions within a variable domain may be viewed as hybrid hypervariable positions in that these positions can be deemed to be within a hypervariable region under one set of criteria while being deemed to be outside a hypervariable region under a different set of criteria. One or more of these positions can also be found in extended hypervariable regions.
• variable domains of native heavy and light chains each contain four framework regions that primarily adopt a ⁇ -sheet configuration, connected by three CDRs, which form loops that connect, and in some cases form part of, the ⁇ - sheet structure.
• the CDRs in each chain are held together in close proximity by the framework regions in the order FR1 -CDR1 -FR2-CDR2-FR3-CDR3-FR4 and, with the CDRs from the other antibody chains, contribute to the formation of the target binding site of antibodies (see Kabat et al., Sequences of Proteins of Immunological Interest, National Institute of Health, Bethesda, MD., 1987).
• numbering of immunoglobulin amino acid residues is performed according to the immunoglobulin amino acid residue numbering system of Kabat et al., unless otherwise indicated (although any antibody numbering scheme, including, but not limited to IMGT and Chothia, can be utilized).
• condition refers to processes by which a patient is prepared for receipt of a transplant containing hematopoietic stem cells.
• a patient may be a hematopoietic stem cell transplant (for instance, as inferred from a sustained increase in the quantity of viable hematopoietic stem cells within a blood sample isolated from a patient following a conditioning procedure and subsequent hematopoietic stem cell transplantation.
• a patient may be a hematopoietic stem cell transplant
• an antibody or antigen-binding fragment thereof capable of binding an antigen expressed by hematopoietic stem cells such as CD1 17 (e.g., GNNK+ CD1 17).
• CD1 17 e.g., GNNK+ CD1 17
• antibody may be covalently conjugated to a cytotoxin so as to form an ADC.
• antigens to a patient in need of hematopoietic stem cell transplant therapy can promote the engraftment of a hematopoietic stem cell graft, for example, by selectively depleting
• conjugate refers to a compound formed by the chemical bonding of a reactive functional group of one molecule, such as an antibody or antigen-binding fragment thereof, with an appropriately reactive functional group of another molecule, such as a cytotoxin described herein.
• Conjugates may include a linker between the two molecules bound to one another.
• linkers that can be used for the formation of a conjugate include peptide-containing linkers, such as those that contain naturally occurring or non-naturally occurring amino acids, such as D-amino acids. Linkers can be prepared using a variety of strategies described herein and known in the art.
• a linker may be cleaved, for example, by enzymatic hydrolysis, photolysis, hydrolysis under acidic conditions, hydrolysis under basic conditions, oxidation, disulfide reduction, nucleophilic cleavage, or organometallic cleavage (see, for example, Leriche et al., Bioorg. Med. Chem., 20:571 -582, 2012).
• the foregoing conjugates are also referred to
• drug antibody conjugate an "antibody drug conjugate” and an “ADC”.
• coupling reaction refers to a chemical reaction in which two or more substituents suitable for reaction with one another react so as to form a chemical moiety that joins
• Coupling reactions include those in which a reactive substituent bound to a fragment that is a cytotoxin, such as a cytotoxin known in the art or described herein, reacts with a suitably reactive substituent bound to a fragment that is an antibody, or antigen-binding fragment thereof, such as an antibody, or antigen-binding fragment thereof, specific for CD1 17 (such as GNNK+ CD1 17) known in the art or described herein.
• suitably reactive substituents include a nucleophile/electrophile pair (e.g., a thiol/haloalkyl pair, an amine/carbonyl pair, or a thiol/a ⁇ -unsatu rated carbonyl pair, among others), a diene/dienophile pair (e.g., an azide/alkyne pair, among others), and the like.
• a nucleophile/electrophile pair e.g., a thiol/haloalkyl pair, an amine/carbonyl pair, or a thiol/a ⁇ -unsatu rated carbonyl pair, among others
• diene/dienophile pair e.g., an azide/alkyne pair, among others
• Coupling reactions include, without limitation, thiol alkylation, hydroxyl alkylation, amine alkylation, amine condensation, amidation, esterification, disulfide formation, cycloaddition (e.g., [4+2] Diels-Alder cycloaddition, [3+2] Huisgen cycloaddition, among others), nucleophilic aromatic substitution, electrophilic aromatic substitution, and other reactive modalities known in the art or described herein.
• cycloaddition e.g., [4+2] Diels-Alder cycloaddition, [3+2] Huisgen cycloaddition, among others
• nucleophilic aromatic substitution e.g., [4+2] Diels-Alder cycloaddition, [3+2] Huisgen cycloaddition, among others
• nucleophilic aromatic substitution e.g., [4+2] Diels-Alder cycloa
• CRU competitive repopulating unit
• drug-to-antibody ratio refers to the number of cytotoxins, e.g., amatoxin, attached to the antibody of an ADC.
• the DAR of an ADC can range from 1 to 8,
• an ADC described herein has a DAR of 1 , 2, 3, 4, 5, 6, 7, or 8.
• the term "donor” refers to a human or animal from which one or more cells are isolated prior to administration of the cells, or progeny thereof, into a recipient.
• the one or more cells may be, for example, a population of hematopoietic stem cells.
• the term "diabody” refers to a bivalent antibody containing two polypeptide chains, in which each polypeptide chain includes V H and V L domains joined by a linker that is too short (e.g., a linker composed of five amino acids) to allow for intramolecular association of V H and V L domains on the same peptide chain. This configuration forces each domain to pair with a linker that is too short (e.g., a linker composed of five amino acids) to allow for intramolecular association of V H and V L domains on the same peptide chain. This configuration forces each domain to pair with a linker that is too short (e.g., a linker composed of five amino acids) to allow for intramolecular association of V H and V L domains on the same peptide chain. This configuration forces each domain to pair with a linker that is too short (e.g., a linker composed of five amino acids) to allow for intramolecular association of V H and V L domains on the same peptid
• the term "triabody” refers to trivalent antibodies containing three peptide chains, each of which contains one V H domain and one V L domain joined by a linker that is exceedingly short (e.g., a linker composed of 1 -2 amino acids) to permit intramolecular association of V H and V L domains within the same peptide chain.
• a linker that is exceedingly short (e.g., a linker composed of 1 -2 amino acids) to permit intramolecular association of V H and V L domains within the same peptide chain.
• peptides configured in this way typically trimerize so as to position the V H and V L domains of neighboring peptide chains spatially proximal to one another (see, for example, Holliger et al., Proc. Natl. Acad. Sci. USA 90:6444-48, 1993).
• a “dual variable domain immunoglobulin” refers to an antibody that combines the target-binding variable domains of two monoclonal antibodies via linkers to create a tetravalent, dual-targeting single agent (see, for example, Gu et al., Meth. Enzymol.,
• the term "endogenous” describes a substance, such as a molecule, cell, tissue, or organ (e.g., a hematopoietic stem cell or a cell of hematopoietic lineage, such as a megakaryocyte, thrombocyte, platelet, erythrocyte, mast cell, myeoblast, basophil, neutrophil, eosinophil, microglial cell, granulocyte, monocyte, osteoclast, antigen-presenting cell, macrophage, dendritic cell, natural killer cell, T-lymphocyte, or B-lymphocyte) that is found naturally in a particular organism, such as a human patient.
• a hematopoietic stem cell or a cell of hematopoietic lineage such as a megakaryocyte, thrombocyte, platelet, erythrocyte, mast cell, myeoblast, basophil, neutrophil, eosinophil, microglial cell, granulocyte, monocyte,
• hematopoietic stem and progenitor cells to repopulate a tissue, whether such cells are naturally circulating or are provided by transplantation.
• the term encompasses all events surrounding or leading up to engraftment, such as tissue homing of cells and colonization of cells within the tissue of interest.
• the engraftment efficiency or rate of engraftment can be evaluated or quantified using any clinically acceptable parameter as known to those of skill in the art and can include, for example, assessment of competitive repopulating units (CRU); incorporation or expression of a marker in tissue(s) into which stem cells have homed, colonized, or become engrafted; or by evaluation of the progress of a subject through disease progression, survival of hematopoietic stem and progenitor cells, or survival of a recipient.
• Engraftment can also be determined by measuring white blood cell counts in peripheral blood during a post-transplant period. Engraftment can also be assessed by measuring recovery of marrow cells by donor cells in a bone marrow aspirate sample.
• exogenous describes a substance, such as a molecule, cell, tissue, or organ (e.g., a hematopoietic stem cell or a cell of hematopoietic lineage, such as a megakaryocyte, thrombocyte, platelet, erythrocyte, mast cell, myeoblast, basophil, neutrophil, eosinophil, microglial cell, granulocyte, monocyte, osteoclast, antigen-presenting cell,
• a hematopoietic stem cell or a cell of hematopoietic lineage such as a megakaryocyte, thrombocyte, platelet, erythrocyte, mast cell, myeoblast, basophil, neutrophil, eosinophil, microglial cell, granulocyte, monocyte, osteoclast, antigen-presenting cell,
• Exogenous substances include those that are provided from an external source to an organism or to cultured matter extracted therefrom.
• frame region includes amino acid residues that are adjacent to the CDRs of an antibody or antigen-binding fragment thereof.
• FW region residues may be present in, for example, human antibodies, humanized antibodies, monoclonal antibodies, antibody fragments, Fab fragments, single chain antibody fragments, scFv fragments, antibody domains, and bispecific antibodies, among others.
• HSCs hematopoietic stem cells
• granulocytes e.g., promyelocytes, neutrophils, eosinophils, basophils
• erythrocytes e.g., reticulocytes, erythrocytes
• thrombocytes e.g., megakaryoblasts, platelet producing megakaryocytes, platelets
• monocytes e.g., monocytes, macrophages
• dendritic cells e.g., NK cells, B-cells and
• Such cells may include CD34 + cells.
• CD34 + cells are immature cells that express the
• CD34 cell surface marker In humans, CD34+ cells are believed to include a subpopulation of cells with the stem cell properties defined above, whereas in mice, HSCs are CD34-. In addition,
• HSCs also refer to long term repopulating HSCs (LT-HSC) and short term repopulating HSCs (ST-HSC).
• LT-HSC long term repopulating HSCs
• ST-HSC short term repopulating HSCs
• LT-HSCs and ST-HSCs are differentiated, based on functional potential and on cell surface marker expression.
• human HSCs are CD34+, CD38-, CD45RA-, CD90+, CD49F+, and lin- (negative for mature lineage markers including CD2, CD3, CD4, CD7, CD8, CD10, CD1 1 B, CD19, CD20, CD56, CD235A).
• bone marrow LT-HSCs are CD34-, SCA-1 +, C- kit+, CD135-, Slamfl/CD150+, CD48-, and lin- (negative for mature lineage markers including Ter1 19, CD1 1 b, Gr1 , CD3, CD4, CD8, B220, IL7ra), whereas ST-HSCs are CD34+, SCA-1 +, C- kit+, CD135-, Slamfl/CD150+, and lin- (negative for mature lineage markers including Ter1 19, CD1 1 b, Gr1 , CD3, CD4, CD8, B220, IL7ra).
• ST-HSCs are less quiescent and more proliferative than LT-HSCs under homeostatic conditions.
• LT-HSC have greater self renewal potential (i.e., they survive throughout adulthood, and can be serially transplanted through successive recipients), whereas ST-HSCs have limited self renewal (i.e., they survive for only a limited period of time, and do not possess serial transplantation potential). Any of these HSCs can be used in the methods described herein.
• ST-HSCs are particularly useful because they are highly proliferative and thus, can more quickly give rise to differentiated progeny.
• hematopoietic stem cell functional potential refers to the functional properties of hematopoietic stem cells which include 1 ) multi-potency (which refers to the ability to differentiate into multiple different blood lineages including, but not limited to, granulocytes (e.g., promyelocytes, neutrophils, eosinophils, basophils), erythrocytes (e.g., reticulocytes, erythrocytes), thrombocytes (e.g., megakaryoblasts, platelet producing
• multi-potency which refers to the ability to differentiate into multiple different blood lineages including, but not limited to, granulocytes (e.g., promyelocytes, neutrophils, eosinophils, basophils), erythrocytes (e.g., reticulocytes, erythrocytes), thrombocytes (e.g., megakaryoblasts, platelet producing
• monocytes e.g., monocytes, macrophages
• dendritic cells e.g., microglia, osteoclasts
• lymphocytes e.g., NK cells, B-cells and T-cells
• human antibody is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences.
• a human antibody may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or during gene rearrangement or by somatic mutation in vivo).
• human antibody as used herein, is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.
• a human antibody can be produced in a human cell (for example, by recombinant expression) or by a non-human animal or a prokaryotic or eukaryotic cell that is capable of expressing functionally rearranged human immunoglobulin (such as heavy chain and/or light chain) genes.
• a human antibody when a human antibody is a single chain antibody, it can include a linker peptide that is not found in native human antibodies.
• an Fv can contain a linker peptide, such as two to about eight glycine or other amino acid residues, which connects the variable region of the heavy chain and the variable region of the light chain. Such linker peptides are considered to be of human origin.
• Human antibodies can be made by a variety of methods known in the art including phage display methods using antibody libraries derived from human immunoglobulin sequences. Human antibodies can also be produced using transgenic mice that are incapable of expressing functional endogenous immunoglobulins, but which can express human immunoglobulin genes (see, for example, PCT Publication Nos. WO 1998/24893; WO 1992/01047; WO 1996/34096; WO 1996/33735; U.S. Patent Nos. 5,413,923; 5,625,126;
• patients that are "in need of" a hematopoietic stem cell transplant include patients that exhibit a defect or deficiency in one or more blood cell types, as well as patients having a stem cell disorder, autoimmune disease, cancer, or other pathology described herein.
• Hematopoietic stem cells generally exhibit 1 ) multi-potency, and can thus differentiate into multiple different blood lineages including, but not limited to, granulocytes (e.g., promyelocytes,
• neutrophils neutrophils, eosinophils, basophils
• erythrocytes e.g., reticulocytes, erythrocytes
• thrombocytes e.g., megakaryoblasts, platelet producing megakaryocytes, platelets
• monocytes e.g., monocytes, macrophages
• dendritic cells microglia, osteoclasts, and lymphocytes
• lymphocytes e.g., NK cells, B-cells and T-cells
• 2) self-renewal and can thus give rise to daughter cells that have equivalent potential as the mother cell, and 3) the ability to be reintroduced into a transplant recipient whereupon they home to the hematopoietic stem cell niche and re-establish productive and sustained hematopoiesis.
• Hematopoietic stem cells can thus be administered to a patient defective or deficient in one or more cell types of the hematopoietic lineage in order to reconstitute the defective or deficient population of cells in vivo.
• the patient may be suffering from cancer, and the deficiency may be caused by administration of a chemotherapeutic agent or other medicament that depletes, either selectively or non-specifically, the cancerous cell population.
• the patient may be suffering from a hemoglobinopathy (e.g., a non-malignant hemoglobinopathy), such as sickle cell anemia, thalassemia, Fanconi anemia, aplastic anemia, and Wiskott-Aldrich syndrome.
• a hemoglobinopathy e.g., a non-malignant hemoglobinopathy
• the subject may be one that is suffering from adenosine deaminase severe combined immunodeficiency (ADA SCID), HIV/AIDS, metachromatic leukodystrophy, Diamond-Blackfan anemia, and Schwachman-Diamond syndrome.
• ADA SCID adenosine deaminase severe combined immunodeficiency
• the subject may have or be affected by an inherited blood disorder (e.g., sickle cell anemia) or an autoimmune disorder.
• the subject may have or be affected by a malignancy, such as neuroblastoma or a hematologic cancer.
• the subject may have a leukemia, lymphoma, or myeloma.
• the subject has acute myeloid leukemia, acute lymphoid leukemia, chronic myeloid leukemia, chronic lymphoid leukemia, multiple myeloma, diffuse large B-cell lymphoma, or non-Hodgkin's lymphoma.
• the subject has myelodysplastic syndrome.
• the subject has an autoimmune disease, such as scleroderma, multiple sclerosis, ulcerative colitis, Crohn's disease, Type 1 diabetes, or another autoimmune pathology described herein.
• the subject is in need of chimeric antigen receptor T-cell (CART) therapy.
• the subject has or is otherwise affected by a metabolic storage disorder.
• the subject may suffer or otherwise be affected by a metabolic disorder selected from the group consisting of glycogen storage diseases, mucopolysaccharidoses, Gaucher's Disease, Hurlers Disease, sphingolipidoses, metachromatic leukodystrophy, or any other diseases or disorders which may benefit from the treatments and therapies disclosed herein and including, without limitation, severe combined immunodeficiency, Wiscott-Aldrich syndrome, hyper immunoglobulin M (IgM) syndrome, Chediak-Higashi disease, hereditary lymphohistiocytosis, osteopetrosis, osteogenesis imperfecta, storage diseases, thalassemia major, sickle cell disease, systemic sclerosis, systemic lupus erythematosus, multiple sclerosis, juvenile rheumatoid arthritis and those diseases, or disorders described in "Bone Marrow Transplantation for Non-Malignant Disease," ASH Education Book, 1 :319-338 (2000), the disclosure of which is incorporated herein by reference in its entirety as
• a patient "in need of" a hematopoietic stem cell transplant may one that is or is not suffering from one of the foregoing pathologies, but nonetheless exhibits a reduced level (e.g., as compared to that of an otherwise healthy subject) of one or more endogenous cell types within the hematopoietic lineage, such as megakaryocytes, thrombocytes, platelets, erythrocytes, mast cells, myeoblasts, basophils, neutrophils, eosinophils, microglia, granulocytes, monocytes, osteoclasts, antigen-presenting cells, macrophages, dendritic cells, natural killer cells, T- lymphocytes, and B- lymphocytes.
• endogenous cell types within the hematopoietic lineage such as megakaryocytes, thrombocytes, platelets, erythrocytes, mast cells, myeoblasts, basophils, neutrophils, eosinophil
• FACS fluorescence activated cell sorting
• the term "recipient” refers to a patient that receives a transplant, such as a transplant containing a population of hematopoietic stem cells.
• a transplant such as a transplant containing a population of hematopoietic stem cells.
• administered to a recipient may be, e.g., autologous, syngeneic, or allogeneic cells.
• sample refers to a specimen (e.g., blood, blood component (e.g., serum or plasma), urine, saliva, amniotic fluid, cerebrospinal fluid, tissue (e.g., placental or dermal), pancreatic fluid, chorionic villus sample, and cells) taken from a subject.
• a specimen e.g., blood, blood component (e.g., serum or plasma), urine, saliva, amniotic fluid, cerebrospinal fluid, tissue (e.g., placental or dermal), pancreatic fluid, chorionic villus sample, and cells
• scFv refers to a single chain Fv antibody in which the variable domains of the heavy chain and the light chain from an antibody have been joined to form one chain.
• scFv fragments contain a single polypeptide chain that includes the variable region of an antibody light chain (V L ) (e.g., CDR-L1 , CDR-L2, and/or CDR-L3) and the variable region of an antibody heavy chain (V H ) (e.g., CDR-H1 , CDR-H2, and/or CDR-H3) separated by a linker.
• V L variable region of an antibody light chain
• V H variable region of an antibody heavy chain
• the linker that joins the V L and V H regions of a scFv fragment can be a peptide linker composed of proteinogenic amino acids.
• linkers can be used to so as to increase the resistance of the scFv fragment to proteolytic degradation (for example, linkers containing D-amino acids), in order to enhance the solubility of the scFv fragment (for example, hydrophilic linkers such as polyethylene glycol-containing linkers or polypeptides containing repeating glycine and serine residues), to improve the biophysical stability of the molecule (for example, a linker containing cysteine residues that form intramolecular or intermolecular disulfide bonds), or to attenuate the immunogenicity of the scFv fragment (for example, linkers containing glycosylation sites).
• linkers containing D-amino acids for example, hydrophilic linkers such as polyethylene glycol-containing linkers or polypeptides containing repeating glycine and serine residues
• hydrophilic linkers such as polyethylene glycol-containing linkers or polypeptides containing repeating glycine and serine residues
• variable regions of the scFv molecules described herein can be modified such that they vary in amino acid sequence from the antibody molecule from which they were derived.
• nucleotide or amino acid substitutions leading to conservative substitutions or changes at amino acid residues can be made (e.g., in CDR and/or framework residues) so as to preserve or enhance the ability of the scFv to bind to the antigen recognized by the corresponding antibody.
• the terms “subject” and “patient” refer to an organism, such as a human, that receives treatment for a particular disease or condition as described herein.
• a patient such as a human patient, may receive treatment prior to hematopoietic stem cell transplant therapy in order to promote the engraftment of exogenous hematopoietic stem cells.
• stem cell disorder broadly refers to any disease, disorder, or condition that may be treated or cured by conditioning a subject's target tissues, and/or by ablating an endogenous stem cell population in a target tissue (e.g., ablating an endogenous hematopoietic stem or progenitor cell population from a subject's bone marrow tissue) and/or by engrafting or transplanting stem cells in a subject's target tissues.
• Type I diabetes has been shown to be cured by hematopoietic stem cell transplant and may benefit from conditioning in accordance with the compositions and methods described herein.
• Additional disorders that can be treated using the compositions and methods described herein include, without limitation, sickle cell anemia, thalassemias, Fanconi anemia, aplastic anemia, Wiskott-Aldrich syndrome, ADA SCID,
• HIV/AIDS metachromatic leukodystrophy
• Diamond-Blackfan anemia and Schwachman-Diamond syndrome.
• Additional diseases that may be treated using the patient conditioning and/or hematopoietic stem cell transplant methods described herein influde inherited blood disorders
• a malignancy such as a neuroblastoma or a hematologic cancer, such as leukemia, lymphoma, and myeloma.
• the cancer may be acute myeloid leukemia, acute lymphoid leukemia, chronic myeloid leukemia, chronic lymphoid leukemia, multiple myeloma, diffuse large B-cell lymphoma, or non- Hodgkin's lymphoma.
• Additional diseases treatable using the conditioning and/or transplantation methods described herein include myelodysplastic syndrome.
• the subject has or is otherwise affected by a metabolic storage disorder.
• the subject may suffer or otherwise be affected by a metabolic disorder selected from the group consisting of glycogen storage diseases, mucopolysaccharidoses, Gaucher's Disease, Hurlers Disease,
• sphingolipidoses metachromatic leukodystrophy, or any other diseases or disorders which may benefit from the treatments and therapies disclosed herein and including, without limitation, severe combined immunodeficiency, Wiscott-Aldrich syndrome, hyper immunoglobulin M (IgM) syndrome, Chediak-Higashi disease, hereditary lymphohistiocytosis, osteopetrosis, osteogenesis imperfecta, storage diseases, thalassemia major, sickle cell disease, systemic sclerosis, systemic lupus erythematosus, multiple sclerosis, juvenile rheumatoid arthritis and those diseases, or disorders described in "Bone Marrow Transplantation for Non-Malignant Disease," ASH Education Book, 1 :319-338 (2000), the disclosure of which is incorporated herein by reference in its entirety as it pertains to pathologies that may be treated by administration of hematopoietic stem cell transplant therapy.
• IgM hyper immunoglobulin M
• transfection refers to any of a wide variety of techniques commonly used for the introduction of exogenous DNA into a prokaryotic or eukaryotic host cell, such as electroporation, lipofection, calcium- phosphate precipitation, DEAE- dextran transfection and the like.
• the terms “treat” or “treatment” refers to reducing the severity and/or frequency of disease symptoms, eliminating disease symptoms and/or the underlying cause of said symptoms, reducing the frequency or likelihood of disease symptoms and/or their underlying cause, and improving or remediating damage caused, directly or indirectly, by disease.
• Beneficial or desired clinical results include, but are not limited to, promoting the engraftment of exogenous hematopoietic cells in a patient following antibody conditioning therapy as described herein and subsequent hematopoietic stem cell transplant therapy. Additional beneficial results include an increase in the cell count or relative concentration of hematopoietic stem cells in a patient in need of a hematopoietic stem cell transplant following conditioning therapy and subsequent
• Beneficial results of therapy described herein may also include an increase in the cell count or relative concentration of one or more cells of hematopoietic lineage, such as a megakaryocyte, thrombocyte, platelet, erythrocyte, mast cell, myeoblast, basophil, neutrophil, eosinophil, microglial cell, granulocyte, monocyte, osteoclast, antigen-presenting cell, macrophage, dendritic cell, natural killer cell, T- lymphocyte, or B-lymphocyte, following conditioning therapy and subsequent hematopoietic stem cell transplant therapy.
• hematopoietic lineage such as a megakaryocyte, thrombocyte, platelet, erythrocyte, mast cell, myeoblast, basophil, neutrophil, eosinophil, microglial cell, granulocyte, monocyte, osteoclast, antigen-presenting cell, macrophage, dendritic cell, natural killer cell, T- lymphocyte, or B-lymphocyte, following conditioning
• Additional beneficial results may include the reduction in quantity of a disease-causing cell population, such as a population of cancer cells (e.g., CD1 17+ leukemic cells) or autoimmune cells (e.g., CD1 17+ autoimmune lymphocytes, such as a CD1 17+ T-cell that expresses a T-cell receptor that cross-reacts with a self antigen).
• a disease-causing cell population such as a population of cancer cells (e.g., CD1 17+ leukemic cells) or autoimmune cells (e.g., CD1 17+ autoimmune lymphocytes, such as a CD1 17+ T-cell that expresses a T-cell receptor that cross-reacts with a self antigen).
• a disease-causing cell population such as a population of cancer cells (e.g., CD1 17+ leukemic cells) or autoimmune cells (e.g., CD1 17+ autoimmune lymphocytes, such as a CD1 17+ T-cell that expresses a T
• variants and “derivative” are used interchangeably and refer to naturally-occurring, synthetic, and semi-synthetic analogues of a compound, peptide, protein, or other substance described herein.
• a variant or derivative of a compound, peptide, protein, or other substance described herein may retain or improve upon the biological activity of the original material.
• vector includes a nucleic acid vector, such as a piasmid, a DNA vector, a piasmid, a RNA vector, virus, or other suitable replicon.
• Expression vectors described herein may contain a polynucleotide sequence as well as, for example, additional sequence elements used for the expression of proteins and/or the integration of these polynucleotide sequences into the genome of a mammalian cell.
• Certain vectors that can be used for the expression of antibodies and antibody fragments of the invention include plasmids that contain regulatory sequences, such as promoter and enhancer regions, which direct gene transcription.
• kits for expression of antibodies and antibody fragments contain polynucleotide sequences that enhance the rate of translation of these genes or improve the stability or nuclear export of the mRNA that results from gene transcription. These sequence elements may include, for example, 5' and 3' untranslated regions and a polyadenylation signal site in order to direct efficient transcription of the gene carried on the expression vector.
• the expression vectors described herein may also contain a polynucleotide encoding a marker for selection of cells that contain such a vector. Examples of a suitable marker include genes that encode resistance to antibiotics, such as ampicillin, chloramphenicol, kanamycin, and nourseothricin.
• R is hydrogen (“aldehyde”), alkyl, alkenyl, alkynyl, carbocyclyl, aryl, heteroaryl, or heterocyclyl, as defined herein.
• Non-limiting examples include formyl, acetyl, propanoyl, benzoyl, and acryloyl.
• alkyl refers to a straight- or branched-chain alkyl group having, for example, from 1 to 20 carbon atoms in the chain.
• alkyl groups include methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, tert-pentyl, hexyl, isohexyl, and the like.
• alkylene refers to a straight- or branched-chain divalent alkyl group. The divalent positions may be on the same or different atoms within the alkyl chain.
• alkylene examples include methylene, ethylene, propylene, isopropylene, and the like.
• heteroalkyl refers to a straight or branched-chain alkyl group having, for example, from 1 to 20 carbon atoms in the chain, and further containing one or more heteroatoms (e.g., oxygen, nitrogen, or sulfur, among others) in the chain.
• heteroalkylene refers to a straight- or branched-chain divalent heteroalkyl group.
• the divalent positions may be on the same or different atoms within the heteroalkyl chain.
• the divalent positions may be one or more heteroatoms.
• alkenyl refers to a straight- or branched-chain alkenyl group having, for example, from 2 to 20 carbon atoms in the chain.
• alkenyl groups include vinyl, propenyl, isopropenyl, butenyl, tert-butylenyl, hexenyl, and the like.
• alkenylene refers to a straight- or branched-chain divalent alkenyl group. The divalent positions may be on the same or different atoms within the alkenyl chain. Examples of alkenylene include ethenylene, propenylene, isopropenylene, butenylene, and the like.
• heteroalkenyl refers to a straight- or branched-chain alkenyl group having, for example, from 2 to 20 carbon atoms in the chain, and further containing one or more heteroatoms (e.g., oxygen, nitrogen, or sulfur, among others) in the chain.
• heteroalkenylene refers to a straight- or branched-chain divalent heteroalkenyl group.
• the divalent positions may be on the same or different atoms within the heteroalkenyl chain.
• the divalent positions may be one or more heteroatoms.
• alkynyl refers to a straight- or branched-chain alkynyl group having, for example, from 2 to 20 carbon atoms in the chain.
• alkynyl groups include propargyl, butynyl, pentynyl, hexynyl, and the like.
• alkynylene refers to a straight- or branched-chain divalent alkynyl group. The divalent positions may be on the same or different atoms within the alkynyl chain.
• heteroalkynyl refers to a straight- or branched-chain alkynyl group having, for example, from 2 to 20 carbon atoms in the chain, and further containing one or more heteroatoms (e.g., oxygen, nitrogen, or sulfur, among others) in the chain.
• heteroalkynylene refers to a straight- or branched-chain divalent heteroalkynyl group.
• the divalent positions may be on the same or different atoms within the heteroalkynyl chain.
• the divalent positions may be one or more heteroatoms.
• cycloalkyi refers to a monocyclic, or fused, bridged, or spiro polycyclic ring structure that is saturated and has, for example, from 3 to 12 carbon ring atoms.
• cycloalkyi groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, bicyclo[3.1 .0]hexane, and the like.
• cycloalkylene refers to a divalent cycloalkyl group. The divalent positions may be on the same or different atoms within the ring structure. Examples of
• cycloalkylene include cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, and the like.
• heterocyloalkyl refers to a monocyclic, or fused, bridged, or spiro polycyclic ring structure that is saturated and has, for example, from 3 to 12 ring atoms per ring structure selected from carbon atoms and heteroatoms selected from, e.g., nitrogen, oxygen, and sulfur, among others.
• the ring structure may contain, for example, one or more oxo groups on carbon, nitrogen, or sulfur ring members.
• heterocycloalkyls include by way of example and not limitation dihydroypyridyl, tetrahydropyridyl (piperidyl), tetrahydrothiophenyl, piperidinyl, 4-piperidonyl, pyrrolidinyl, 2-pyrrolidonyl, tetrahydrofuranyl, tetrahydropyranyl, bis- tetrahydropyranyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl,
• heterocycloalkylene refers to a divalent heterocyclolalkyl group.
• the divalent positions may be on the same or different atoms within the ring structure.
• aryl refers to a monocyclic or multicyclic aromatic ring system containing, for example, from 6 to 19 carbon atoms.
• Aryl groups include, but are not limited to, phenyl, fluorenyl, naphthyl, and the like. The divalent positions may be one or more heteroatoms.
• arylene refers to a divalent aryl group.
• the divalent positions may be on the same or different atoms.
• heteroaryl refers to a monocyclic heteroaromatic, or a bicyclic or a tricyclic fused-ring heteroaromatic group in which one or more ring atoms is a heteroatom, e.g., nitrogen, oxygen, or sulfur.
• Heteroaryl groups include pyridyl, pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, 1 ,2,3-triazolyl, 1 ,2,4-triazolyl, 1 ,2,3- oxadiazolyl, 1 ,2,4-oxadia-zolyl, 1 ,2,5-oxadiazolyl, 1 ,3,4-oxadiazolyl, 1 ,3,4-triazinyl, 1 ,2,3-triazinyl, benzofuryl, [2,3-dihydro]benzofuryl, isobenzofuryl, benzothienyl, benzotriazolyl, isobenzothienyl, indolyl, isoindolyl, 3H-indolyl, benzimidazolyl, imidazo[1
• heteroarylene refers to a divalent heteroaryl group.
• the divalent positions may be on the same or different atoms.
• the divalent positions may be one or more heteroatoms.
• alkenylene "heteroalkenyl”, “heteroalkenylene”, “alkynyl”, “alkynylene”, “heteroalkynyl”,
• heteroalkynylene "cycloalkyl”, “cycloalkylene”, “heterocyclolalkyl”, heterocycloalkylene", "aryl,”
• arylene, “heteroaryl”, and “heteroarylene” groups can optionally be substituted with, for example, from 1 to 5 substituents selected from the group consisting of alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, alkyl aryl, alkyl heteroaryl, alkyl cycloalkyl, alkyl heterocycloalkyl, amino, ammonium, acyl, acyloxy, acylamino, aminocarbonyl, alkoxycarbonyl, ureido, carbamate, aryl, heteroaryl, sulfinyl, sulfonyl, alkoxy, sulfanyl, halogen, carboxy, trihalomethyl, cyano, hydroxy, mercapto, nitro, and the like.
• substitution may include situations in which neighboring substituents have undergone ring closure, such as ring closure of vicinal functional substituents, to form, for instance, lactams, lactones, cyclic anhydrides, acetals, hemiacetals, thioacetals, aminals, and hemiaminals, formed by ring closure, for example, to furnish a protecting group.
• ring closure such as ring closure of vicinal functional substituents, to form, for instance, lactams, lactones, cyclic anhydrides, acetals, hemiacetals, thioacetals, aminals, and hemiaminals, formed by ring closure, for example, to furnish a protecting group.
• radical naming conventions can include either a mono- radical or a di-radical, depending on the context.
• a substituent requires two points of attachment to the rest of the molecule, it is understood that the substituent is a di-radical.
• a substituent identified as alkyl that requires two points of attachment includes di- radicals such as -CH 2 -, -CH 2 CH 2 -, -CH 2 CH(CH 3 )CH 2 -, and the like.
• Other radical naming conventions clearly indicate that the radical is a di-radical such as "alkylene,” “alkenylene,” “arylene,” “heterocycloalkylene,” and the like.
• the present invention is based in part on the discovery of novel anti-CD1 17 antibodies and antigen binding portions thereof that are useful for therapeutic purposes.
• the present invention is also based in part on the discovery that antibodies, or antigen-binding fragments thereof, capable of binding CD1 17, such as GNNK+ CD1 17, can be used as therapeutic agents alone or as ADCs to (i) treat cancers (such as acute myelogenous leukemia or myelodysplastic syndrome) and autoimmune diseases characterized by CD1 17+ cells and (ii) promote the engraftment of transplanted hematopoietic stem cells in a patient in need of transplant therapy.
• therapeutic activities can be caused, for instance, by the binding of anti-CD1 17 antibodies, or antigen-binding fragments thereof, to CD1 17 (e.g., GNNK+ CD1 17) expressed on the surface of a cell, such as a cancer cell, autoimmune cell, or hematopoietic stem cell and subsequently inducing cell death.
• CD1 17 e.g., GNNK+ CD1 17
• the depletion of endogenous hematopoietic stem cells can provide a niche toward which transplanted hematopoietic stem cells can home, and subsequently establish productive hematopoiesis. In this way, transplanted hematopoietic stem cells may successfully engraft in a patient, such as human patient suffering from a stem cell disorder described herein.
• Antibodies and antigen-binding fragments capable of binding human CD1 17 can be used in conjunction with the compositions and methods described herein in order to condition a patient for
• CD1 17 Polymorphisms affecting the coding region or extracellular domain of CD1 17 in a significant percentage of the population are not currently well- known in non-oncology indications.
• Two of the CD1 17 isoforms are located on the intracellular domain of the protein, and two are present in the external juxtamembrane region.
• the two extracellular isoforms, GNNK+ and GNNK- differ in the presence (GNNK+) or absence (GNNK-) of a 4 amino acid sequence.
• GNNK+ isoform can be used as an immunogen in order to generate antibodies capable of binding CD1 17, as antibodies generated against this isoform will be inclusive of the GNNK+ and GNNK- proteins.
• anti-CD1 17 antibodies whose heavy and light chain amino acid sequences are provided in Table 1 , Table 6, Table 8, and Table 16.
• anti-CD1 17 antibody drug conjugates comprising binding regions (heavy and light chain CDRs or variable regions) as set forth in SEQ ID Nos: 7 to 168.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 8.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 9. In one embodiment, the anti-CD1 17 antibody, or antigen binding portion thereof, comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 1 1 .
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 12.
• the anti- CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 13.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 14.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 16.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 17.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 17.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 17.
• CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 18.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 19.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 21 .
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 22.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 22.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 22.
• CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 23.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 24, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 25.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 28, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 29.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 30, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 31 .
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 32, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 33.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 34, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO:35.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 36, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 37. In one embodiment, the anti-CD1 17 antibody, or antigen binding portion thereof, comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO:
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 32, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 42.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 43, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 44.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 45, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 46.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 47, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 48.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO:
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 53, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 54.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 55, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 56.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 57, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 58.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 9, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 60.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO:
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 64, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 65.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 66, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 67.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 68, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 69. In one embodiment, the anti-CD1 17 antibody, or antigen binding portion thereof, comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 70, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 71 . In one embodiment, the anti-CD1 17 antibody, or antigen binding portion thereof, comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO:
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 76, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 77.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 78, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 79.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 80, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 81 .
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 82, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 83.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO:
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 88.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 89.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 90. In one embodiment, the anti-CD1 17 antibody, or antigen binding portion thereof, comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 91 . In one embodiment, the anti-CD1 17 antibody, or antigen binding portion thereof, comprises a heavy chain variable region as set forth in the amino acid sequence of
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 93.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 94.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 94.
• CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO:95.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 96.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 97. In one embodiment, the anti-CD1 17 antibody, or antigen binding portion thereof, comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 97.
• the anti- CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 143, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 144.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 151 , and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 152.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 143, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 156.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 158, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 156.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 160, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 152.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 98, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 99.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 99.
• the anti- CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 7, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 100.
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 98, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 101 .
• the anti-CD1 17 antibody, or antigen binding portion thereof comprises a heavy chain variable region as set forth in the amino acid sequence of SEQ ID NO: 98, and a light chain variable region as set forth in the amino acid sequence of SEQ ID NO: 102.
• the antibody is an intact antibody comprising a heavy chain and a light chain variable region as set forth in Table 1 .
• the anti-CD1 17 antibody is engineered to have a short half life.
• nucleic acid sequences corresponding to the heavy and light chain regions of certain sequences described above are provided in Table 2.

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