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WO2025111450A1 - Conjugués anticorps anti-cd74-médicament et leurs procédés d'utilisation - Google Patents

Conjugués anticorps anti-cd74-médicament et leurs procédés d'utilisation Download PDF

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Publication number
WO2025111450A1
WO2025111450A1 PCT/US2024/056875 US2024056875W WO2025111450A1 WO 2025111450 A1 WO2025111450 A1 WO 2025111450A1 US 2024056875 W US2024056875 W US 2024056875W WO 2025111450 A1 WO2025111450 A1 WO 2025111450A1
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Prior art keywords
cancer
antibody
seq
antigen
heavy chain
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Inventor
Matthew T. Burger
Zhuoliang Chen
Joseph Anthony D'ALESSIO
Eric Andrew Mcneill
Katsumasa Nakajima
Richard Vaughan Newcombe
Bing Yu
Qiang Zhang
Ana Leticia MARAGNO
Jerôme-Benoit STARCK
Imre Fejes
Zoltan Szlavik
Stuart Ray
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Novartis AG
Les Laboratoires Servier SAS
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Novartis AG
Les Laboratoires Servier SAS
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Publication of WO2025111450A1 publication Critical patent/WO2025111450A1/fr
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—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 organic compound
    • A61K47/55—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 organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6849—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—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
    • A61K47/6851—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 determinant of a tumour cell
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6889—Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • ADCs antibody-drug conjugates
  • the disclosure further relates to methods and compositions useful in the treatment and/or diagnosis of cancers that express a target antigen and/or are amenable to treatment by modulating expression and/or activity of Bcl-2 family proteins, as well as methods of making those compositions.
  • Apoptosis (programmed cell death) is an evolutionarily conserved pathway essential for tissue homeostasis, development and removal of damaged cells. Deregulation of apoptosis contributes to human diseases, including malignancies, neurodegenerative disorders, diseases of the immune system and autoimmune diseases (Hanahan and Weinberg, Cell.
  • the Bcl-2 protein family comprises key regulators of cell survival which can suppress (e.g., Bcl-2, Bcl-xL, Mcl-1) or promote (e.g., Bad, Bax) apoptosis (Gross et al., Genes Dev.1999 Aug 1;13(15):1899-911, Youle and Strasser, Nat. Rev. Mol. Cell Biol.2008 Jan;9(1):47-59). 1 ME150999036v.1 132043-01120 [06] In the face of stress stimuli, whether a cell survives or undergoes apoptosis is dependent on the extent of pairing between the Bcl-2 family members that promote cell death with family members that promote cell survival.
  • Bcl-2 homology 3 (BH3) domain of proapoptotic family members into a groove on the surface of pro- survival members.
  • Bcl-2 homology (BH) domain defines the membership of the Bcl- 2 family, which is divided into three main groups depending upon the particular BH domains present within the protein.
  • the prosurvival members such as Bcl-2, Bcl-xL, and Mcl-1 contain BH domains 1–4, whereas Bax and Bak, the proapoptotic effectors of mitochondrial outer membrane permeabilization during apoptosis, contain BH domains 1–3 (Youle and Strasser, Nat. Rev. Mol. Cell Biol.2008 Jan;9(1):47-59).
  • Bcl- xL (also named BCL2L1, from BCL2-like 1) is frequently amplified in cancer (Beroukhim et al., Nature 2010 Feb 18;463(7283):899-905) and it has been shown that its expression inversely correlates with sensitivity to more than 120 anti-cancer therapeutic molecules in a representative panel of cancer cell lines (NCI-60) (Amundson et al., Cancer Res.2000 Nov 1;60(21):6101-10).
  • This new class of drugs includes inhibitors of Bcl-2, Bcl-xL, Bcl-w and Mcl-1.
  • the first BH3 mimetics described were ABT- 737 and ABT-263, targeting Bcl-2, Bcl-xL and Bcl-w (Park et al., J. Med. Chem. 2008 Nov 13;51(21):6902-15; Roberts et al., J. Clin. Oncol. 2012 Feb 10;30(5):488-96).
  • selective inhibitors of Bcl-2 (ABT-199 and S55746 – Souers et al., Nat Med.
  • ABT-263 has shown activity in several hematological malignancies and solid tumors (Shoemaker et al., Clin. Cancer Res.2008 Jun 1;14(11):3268-77; Ackler et al., Cancer Chemother. Pharmacol.2010 Oct;66(5):869-80; Chen et al., Mol. Cancer Ther.2011 Dec;10(12):2340-9).
  • ABT-263 exhibited objective antitumor activity in lymphoid malignancies (Wilson et al., Lancet Oncol.2010 Dec;11(12):1149-59; Roberts et al., J. Clin. Oncol.
  • mice treated with ABT-737 resulted in potent inhibition of lymphocyte proliferation in vitro.
  • mice treated with ABT-737 in animal models of arthritis and lupus showed a significant decrease in disease severity (Bardwell et al., J Clin Invest. 1997 Feb 1;99(3):439-46. doi: 10.1172/JCI119178.PMID: 9022077).
  • CD74 (DHLAG) is an established and attractive target for antibody drug conjugates due to its restricted expression on normal tissues and significant upregulation in a range of hematological malignancies. CD74 functions as a chaperone that is necessary for the assembly and trafficking of MHC class II complexes as well as a receptor for macrophage migration inhibitory receptor (MIF).
  • MIF macrophage migration inhibitory receptor
  • CD74 is significantly upregulated at both the RNA and protein level in a range of B-cell and myeloid cell malignancies including acute myeloid leukemia (AML), multiple myeloma, and diffuse large B-cell lymphoma. Additionally CD74 is known to rapidly internalize upon antibody engagement and traffic to the lysosome as well as to be rapidly repopulated on the surface of tumor cells following internalization. Antibodies and antibody drug conjugates targeting CD74 have been shown previously to demonstrate anti-tumor activity in preclinical models of cancer.
  • the present disclosure provides an antibody-drug conjugate of Formula (I), (I), wherein is selected from the group consisting of: 4 ME150999036v.1 132043-01120 5 ME150999036v.1 132043-01120 wherein indicates the point of attachment to Ab; wherein Ab is an anti-CD74 antibody or an antigen-binding fragment thereof comprising three heavy chain complementarity determining regions (CDRs) and three light chain CDRs selected from the group consisting of: 1) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:1, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:2, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:10, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:11, and a light chain CDR3 (LCDR3) consisting of SEQEQ
  • the present disclosure provides an antibody-drug conjugate, wherein the anti-CD74 antibody or antigen-binding fragment thereof comprises: a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:17, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:18.
  • the definitions of the remaining variables are provided in any other embodiments described herein.
  • the present disclosure provides an antibody-drug conjugate, wherein the anti-CD74 antibody or antigen-binding fragment thereof comprises: a heavy chain amino acid sequence of SEQ ID NO:19 or a sequence that is at least 95% identical to SEQ ID NO:19, and a light chain amino acid sequence of SEQ ID NO:20 or a sequence that is at least 95% identical to SEQ ID NO:20.
  • the definitions of the remaining variables are provided in any other embodiments described herein.
  • the present disclosure provides an antibody-drug conjugate, wherein the anti-CD74 antibody or antigen-binding fragment thereof comprises one or more Fc silencing mutations. The definitions of the remaining variables are provided in any other embodiments described herein.
  • the present disclosure provides an antibody-drug conjugate, wherein the anti-CD74 antibody or antigen-binding fragment thereof comprise one or more CysMab mutations.
  • the definitions of the remaining variables are provided in any other embodiments described herein.
  • the present disclosure provides an antibody-drug conjugate, wherein the anti-CD74 antibody or antigen-binding fragment thereof comprises one or more CysMab mutations selected from E152C, S375C, or both E152C and S375C of the heavy chain of the antibody or antigen binding fragment thereof, wherein the position is numbered according to the EU system.
  • the definitions of the remaining variables are provided in any other embodiments described herein.
  • the present disclosure provides an antibody-drug conjugate, wherein 7 ME150999036v.1 132043-01120 the anti-CD74 antibody or antigen-binding fragment thereof comprises a Fab fragment, a Fab′ fragment, a F(ab′)2 fragment, a single chain Fv (scFv), a disulfide-linked Fv (sdFv), a Fd fragment, a Fv fragment, a dAb fragment, a maxibody, a minibody, an intrabody, a diabody, a half antibody, or a one-arm antibody.
  • the definitions of the remaining variables are provided in any other embodiments described herein.
  • the present disclosure provides an antibody-drug conjugate, wherein the anti-CD74 antibody or antigen-binding fragment thereof is a monoclonal antibody. The definitions of the remaining variables are provided in any other embodiments described herein. [21] In another embodiment, the present disclosure provides an antibody-drug conjugate, wherein The definitions of the remaining variables are provided in any other embodiments described herein. [22] In another embodiment, the present disclosure provides an antibody-drug conjugate of, wherein is: The definitions of the remaining variables are provided in any other embodiments described herein. [23] In another embodiment, the present disclosure provides an antibody-drug conjugate, wherein 8 ME150999036v.1 132043-01120 .
  • the present disclosure provides an antibody-drug conjugate of, wherein is: The definitions of the remaining variables are provided in any other embodiments described herein. [25] In another embodiment, the present disclosure provides an antibody-drug conjugate, wherein 9 ME150999036v.1 132043-01120 . The definitions of the remaining variables are provided in any other embodiments described herein. [26] In another embodiment, the present disclosure provides an antibody-drug conjugate, wherein . The definitions of the remaining variables are provided in a any other embodiment described herein. [27] In another embodiment, the present disclosure provides an antibody-drug conjugate of an antibody-drug conjugate, wherein is: .
  • a is an integer from 1 to 8. In some embodiments, a is an integer from 1 to 5. In some embodiments, a is an integer from 2 to 4. In some embodiments, a is 2. In some 10 ME150999036v.1 132043-01120 embodiments, a is 4. In some embodiments, a is determined by liquid chromatography-mass spectrometry (LC-MS).
  • LC-MS liquid chromatography-mass spectrometry
  • the anti-CD74 antibody is VHmil x VK1aNQ.
  • the antibody or antigen-binding fragment binds to a target antigen on a cancer cell.
  • the target antigen is CD74.
  • the present disclosure provides an antibody or antigen-binding fragment comprising one or more SEQ IDs listed in Tables A2 and A3 described herein.
  • compositions comprising multiple copies of an antibody-drug conjugate (e.g., any of the exemplary antibody-drug conjugates described herein).
  • the average a of the antibody-drug conjugates in the composition is from about 1 to about 8, about 1 to about 6, about 1 to about 4, about 1 to about 2, or about 2 to about 4.
  • compositions comprising an antibody-drug conjugate (e.g., any of the exemplary antibody-drug conjugates described herein) or a composition (e.g., any of the exemplary compositions described herein), and a pharmaceutically acceptable carrier.
  • an antibody-drug conjugate e.g., any of the exemplary antibody-drug conjugates described herein
  • a composition e.g., any of the exemplary compositions described herein
  • a pharmaceutically acceptable carrier e.g., any of the exemplary compositions described herein
  • therapeutic uses for the described antibody-drug conjugate compounds and compositions e.g., in treating a cancer.
  • the present disclosure provides methods of treating a cancer (e.g., a cancer that expresses an antigen targeted by the antibody or antigen-binding fragment of the antibody-drug conjugate, such as CD74).
  • the present disclosure provides methods of reducing or slowing the expansion of a cancer cell population in a subject. In some embodiments, the present disclosure provides methods of determining whether a subject having or suspected of having a cancer will be responsive to treatment with an antibody-drug conjugate compound or composition disclosed herein.
  • An exemplary embodiment is a method of treating a subject having or suspected of having a cancer, comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein).
  • the cancer expresses a target antigen. In some embodiments, the target antigen is CD74.
  • the cancer is a tumor or a hematological cancer.
  • the cancer is a breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular cancer, lymphoblastic leukemia including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myelogenous leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, 11 ME150999036v.1 132043-01120 pancreatic cancer, stomach cancer, colon cancer, or head and neck cancer.
  • the cancer is acute myeloid leukemia, multiple myeloma, or B-cell lymphoma.
  • Another exemplary embodiment is a method of reducing or inhibiting the growth of a tumor in a subject, comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein).
  • the tumor expresses a target antigen.
  • the target antigen is CD74.
  • the tumor is a breast cancer, gastric cancer, bladder cancer, brain cancer, cervical cancer, colorectal cancer, esophageal cancer, hepatocellular cancer, melanoma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, pancreatic cancer, stomach cancer, colon cancer, head and neck cancer, or spleen cancer.
  • administration of the antibody-drug conjugate, composition, or pharmaceutical composition reduces or inhibits the growth of the tumor by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%.
  • Another exemplary embodiment is a method of reducing or inhibiting a hematological cancer in a subject, comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein).
  • the tumor expresses a target antigen.
  • the target antigen is CD74.
  • the hematological cancer is chronic lymphocytic leukemia (CLL), follicular lymphoma, mantle cell lymphoma, diffuse large B-cell lymphoma, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myelomonocytic leukemia (CMML), acute monocytic leukemia (AMoL), Hodgkin's lymphoma, non-Hodgkin's lymphoma or myelodysplasia syndrome (MDS).
  • CLL chronic lymphocytic leukemia
  • ALL acute lymphoblastic leukemia
  • AML acute myeloid leukemia
  • CLL chronic lymphocytic leukemia
  • CML chronic myelogenous leukemia
  • CML chronic myelomonocytic leukemia
  • AoL acute monocytic leukemia
  • administration of the antibody-drug conjugate, composition, or pharmaceutical composition reduces or inhibits the growth of the hematological cancer by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%.
  • Another exemplary embodiment is a method of reducing or slowing the expansion of a cancer cell population in a subject, comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein).
  • the cancer cell population expresses a target antigen.
  • the target antigen is CD74.
  • the cancer cell population is from a tumor or a hematological cancer.
  • the cancer cell population is from a breast 12 ME150999036v.1 132043-01120 cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular cancer, lymphoblastic leukemia including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T- cell or B-cell origin, melanoma, myelogenous leukemia, myeloma, oral cancer, ovarian cancer, non- small cell lung cancer, prostate
  • the cancer cell population is from acute myeloid leukemia, multiple myeloma, or B-cell lymphoma.
  • administration of the antibody-drug conjugate, composition, or pharmaceutical composition reduces the cancer cell population by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%.
  • administration of the antibody-drug conjugate, composition, or pharmaceutical composition slows the expansion of the cancer cell population by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%.
  • Another exemplary embodiment is an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein) for use in treating a subject having or suspected of having a cancer.
  • the cancer expresses a target antigen.
  • the target antigen is CD74.
  • the cancer is a tumor or a hematological cancer.
  • the cancer is a breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular cancer, lymphoblastic leukemia including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myelogenous leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, stomach cancer, colon cancer, or head and neck cancer.
  • the cancer is acute myeloid leukemia, multiple myeloma, or B-cell lymphoma.
  • Another exemplary embodiment is a use of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein) in treating a subject having or suspected of having a cancer.
  • the cancer expresses a target antigen.
  • the target antigen is CD74.
  • the cancer is a tumor or a hematological cancer.
  • the cancer is a breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, 13 ME150999036v.1 132043-01120 hepatocellular cancer, lymphoblastic leukemia including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myelogenous leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, stomach cancer, colon cancer, or head and neck cancer.
  • lymphoblastic leukemia including acute lymphoblastic leukemia, follicular lymphoma, lymphoi
  • the cancer is acute myeloid leukemia, multiple myeloma, or B-cell lymphoma.
  • Another exemplary embodiment is a use of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein) in a method of manufacturing a medicament for treating a subject having or suspected of having a cancer.
  • the cancer expresses a target antigen.
  • the target antigen is CD74.
  • the cancer is a tumor or a hematological cancer.
  • the cancer is a breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular cancer, lymphoblastic leukemia including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B- cell origin, melanoma, myelogenous leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, stomach cancer, colon cancer, or head and neck cancer.
  • the cancer is acute myeloid leukemia, multiple myeloma, or B-cell lymphoma.
  • Another exemplary embodiment is a method of determining whether a subject having or suspected of having a cancer will be responsive to treatment with an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein) by providing a biological sample from the subject; contacting the sample with the antibody-drug conjugate; and detecting binding of the antibody-drug conjugate to cancer cells in the sample.
  • the cancer cells in the sample express a target antigen.
  • the cancer expresses a target antigen.
  • the target antigen is CD74.
  • the cancer is a tumor or a hematological cancer.
  • the cancer is a breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular cancer, lymphoblastic leukemia including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myelogenous leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, stomach cancer, colon cancer, or head and neck cancer.
  • the cancer is acute myeloid leukemia, multiple myeloma, or B-cell lymphoma.
  • the sample is a tissue biopsy sample, a blood sample, or a bone marrow sample. 14 ME150999036v.1 132043-01120 [43] Methods of producing the described antibody-drug conjugate compounds and compositions (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein) are also disclosed.
  • An exemplary embodiment is a method of producing an antibody-drug conjugate by conjugating an anti-CD74 antibody or antigen-binding fragment to a dual linker joined or covalently attached to two antineoplastic compounds described herein under conditions that allow conjugation.
  • FIG. 1 depicts a complex structure of VHmil x Vk1aNQ Fab and CD74 peptide showing the Fab as ribbon representation and the bound peptide as sticks with its molecular surface (Connolly surface). The heavy chain is shown in dark grey, the light chain is shown in white and the peptide is colored in light grey. The terminal amino acids of the peptide chain are labelled.
  • FIG. 2 is an unbiased Fo-Fc omit map contoured at 4.4 ⁇ which is shown with the final refined CD74 peptide model as ball-and-stick representation superimposed. Only for the eight C-terminal residues (AA289 to AA296) strong and clear electron density was revealed.
  • FIG. 3 is a graph showing the number of direct intermolecular contacts (# of non-H mAB atoms within 3.8 ⁇ and the reduction in solvent accessible surface ( ⁇ 2 ) upon binding plotted per CD74-peptide amino acid (X-axis).
  • FIG. 4A is a graph showing the number of direct intermolecular contacts (# of non-H mAB atoms within 3.8 ⁇ and the reduction in solvent accessible surface ( ⁇ 2 )) upon binding plotted per amino acid from the VHmil x Vk1aNQ Fab heavy chain (SEQ ID NO:24) (X-axis).
  • FIG. 48 is a graph showing the number of direct intermolecular contacts (# of non-H mAB atoms within 3.8 ⁇ and the reduction in solvent accessible surface ( ⁇ 2 ) upon binding plotted per amino acid from the VHmil x Vk1aNQ Fab heavy chain (SEQ ID NO:24) (X-axis).
  • FIG. 4B is a graph showing the number of direct intermolecular contacts (# of non-H mAB atoms within 3.8 ⁇ and the reduction in solvent accessible surface ( ⁇ 2 )) upon binding plotted per amino acid from the VHmil x Vk1aNQ Fab light chain (SEQ ID NO: 25) (X-axis)).
  • FIG. 5 is a graph showing growth inhibition (GI50) of the CD74 (VHmil x Vk1aNQ) -P1- L12-P4 ADC in ⁇ M for the representative cell lines tested.
  • FIG. 5 is a graph showing growth inhibition (GI50) of the CD74 (VHmil x Vk1aNQ) -P1- L12-P4 ADC in ⁇ M for the representative cell lines tested.
  • FIGS. 6 are graphs showing dose response curves of eight ADCs - VHmil x VK1aNQ-P1-L12- P3, VHmil x VK1aNQ-P1-L17-P2, VHmil x VK1aNQ-P1-L12-P2, VHmil x VK1aNQ-P1-L17-P3, VHmil x VK1aNQ-P1-L17-P4, VHmil x VK1aNQ-P1-L19-P2, VHmil x VK1aNQ-P1-L12-P4, IgG- P1-L12-P4 in the Nomo1, EOL1, and Monomac1 cell lines.
  • Fig. 7A-7D are graphs showing efficacy data generated in a disseminated EOL1-Luc tumor model treated with a CD74-MCL-1/BCL-2 dual ADC (VHmil x VK1aNQ-P1-L12-P4 ADC). Mice were also treated with an isotype control ADC (IgG-P1-L12-P4 ADC). ADCs were administered on Q2Wx2 schedule. EOL-1 tumors Leukemic burden was evaluated using a Xenogen bioluminescence imager. Group averages are presented in (7A), and individual tumor data is presented in (7B-7D).
  • compositions and methods may be understood more readily by reference to the following detailed description taken in connection with the accompanying figures, which form a part of this disclosure.
  • the descriptions refer to compositions and methods of using the compositions. Where the disclosure describes or claims a feature or embodiment associated with a composition, such a feature or embodiment is equally applicable to the methods of using the composition. Likewise, where the disclosure describes or claims a feature or embodiment associated with a method of using a composition, such a feature or embodiment is equally applicable to the composition. [54] When a range of values is expressed, it includes embodiments using any particular value within the range.
  • antibody drug conjugates can be identified using a naming convention in the general format of “target antigen/antibody-payload-dual linker-payload.”
  • the term “P#-L#-P#” refers to a specific dual linker-drug disclosed herein, wherein each of the codes “P#” refers to a specific antineoplastic compound (e.g., a BH3 mimetic) described here unless otherwise specified and L# refers to a specific dual linker unless otherwise specified.
  • an antibody drug conjugate is referred to as “Target X-P1-L1-P2”
  • such a conjugate would 16 ME150999036v.1 132043-01120 comprise an antibody that binds Target X, a dual linker designated as L1, and two payloads designated as P1 and P2, respectively, including an enantiomer, diastereoisomer, atropisomer, deuterated derivative, and/or pharmaceutically acceptable salt of any of the foregoing.
  • an antibody drug conjugate is referred to as “anti-Target X-P1-L1-P2”, such a conjugate would comprise an antibody that binds Target X, a dual linker designated as L1, and two payloads designated as P1 and P2, respectively.
  • an antibody drug conjugate is referred to as “AbX-P1-L1-P2”, such a conjugate would comprise the antibody designated as AbX, a dual linker designated as L1, and two payloads designated as P1 and P2, respectively.
  • a control antibody drug conjugate comprising a non-specific, isotype control antibody may be referenced as “isotype control IgG1-P1-L1-P2” or “IgG1-P1-L1-P2”.
  • Exemplary embodiments of the present disclosure include VHmil x VK1aNQ-P1-L12-P2, VHmil x VK1aNQ-P1-L12-P3, VHmil x VK1aNQ-P1-L12- P4, VHmil x VK1aNQ-P1-L17-P2, VHmil x VK1aNQ-P1-L17-P3, VHmil x VK1aNQ-P1-L17-P4, and VHmil x VK1aNQ-P1-L19-P2.
  • any formula given herein is also intended to represent unlabeled forms as well as isotopically labeled forms of the compounds.
  • Isotopically labeled compounds have structures depicted by the formulae given herein except that one or more atoms are replaced by an atom having a selected atomic mass or mass number.
  • Isotopes that can be incorporated into compounds of the invention include, for example, isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, and chlorine, such as 3 H, 11 C, 13 C, 14 C, 15 N, 18 F, and 36 Cl.
  • the present disclosure includes compounds that incorporate one or more of any of the aforementioned isotopes, including for example, radioactive isotopes, such as 3 H and 14 C, or those into which non-radioactive isotopes, such as 2 H and 13 C are present.
  • isotopically labelled compounds are useful in metabolic studies (with 14 C), reaction kinetic studies (with, for example 2 H or 3 H), detection or imaging techniques, such as positron emission tomography (PET) or single-photon emission computed tomography (SPECT) including drug or substrate tissue distribution assays, or in radioactive treatment of patients.
  • PET positron emission tomography
  • SPECT single-photon emission computed tomography
  • an 18 F or labeled compound may be particularly desirable for PET or SPECT studies.
  • Isotopically-labeled compounds can generally be prepared by conventional techniques known to those skilled in the art, e.g., using an appropriate isotopically-labeled reagents in place of the non-labeled reagent previously employed.
  • Definitions [59] Various terms relating to aspects of the description are used throughout the specification and claims. Such terms are to be given their ordinary meaning in the art unless otherwise indicated. Other specifically defined terms are to be construed in a manner consistent with the definitions provided herein. [60] As used herein, the singular forms “a,” “an,” and “the” include plural forms unless the context clearly dictates otherwise.
  • the term “about” refers to a range of values which are 1% more or less than the specified value.
  • the terms “antibody-drug conjugate,” “antibody conjugate,” “conjugate,” “immunoconjugate,” and “ADC” are used interchangeably, and refer to one or more therapeutic compounds described herein that is linked to one or more anti-CD74 antibodies or antigen-binding fragments.
  • “2a” refers to the number of antineoplastic payloads described herein linked to the antibody or antigen-binding fragment.
  • antibody is used in the broadest sense to refer to an immunoglobulin molecule that recognizes and specifically binds to a target, such as a protein, polypeptide, carbohydrate, polynucleotide, lipid, or combinations of the foregoing through at least one antigen recognition site within the variable region of the immunoglobulin molecule.
  • An antibody can be polyclonal or monoclonal, multiple or single chain, or an intact immunoglobulin, and may be derived from natural sources or from recombinant sources.
  • An “intact” antibody is a glycoprotein that typically comprises at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds.
  • Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region.
  • the heavy chain constant region comprises three domains, CH1, CH2 and CH3.
  • Each light chain is comprised of a light chain variable region (abbreviated herein as 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 18 ME150999036v.1 132043-01120 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 may 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 (C1q) of the classical complement system.
  • An antibody can be a monoclonal antibody, human antibody, humanized antibody, camelised antibody, or chimeric antibody.
  • the antibodies can be of any isotype (e.g., IgG, IgE, IgM, IgD, IgA and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2), or subclass.
  • An antibody can be an intact antibody or an antigen-binding fragment thereof.
  • the antibody or antibody fragment disclosed herein include modified or engineered amino acid residues, e.g., one or more cysteine residues, as sites for conjugation to a drug moiety (Junutula JR, et al., Nat Biotechnol 2008, 26:925-932).
  • the disclosure provides a modified antibody or antibody fragment comprising a substitution of one or more amino acids with cysteine at the positions described herein.
  • Sites for cysteine substitution are in the constant regions of the antibody or antibody fragment and are thus applicable to a variety of antibody or antibody fragment, and the sites are selected to provide stable and homogeneous conjugates.
  • a modified antibody or fragment can have one, two or more cysteine substitutions, and these substitutions can be used in combination with other modification and conjugation methods as described herein.
  • a modified antibody comprises a substitution of one or more amino acids with cysteine on its constant region selected from positions 117, 119, 121, 124, 139, 152, 153, 155, 157, 164, 169, 171, 174, 189, 191, 195, 197, 205, 207, 246, 258, 269, 274, 286, 288, 290, 292, 293, 320, 322, 326, 333, 334, 335, 337, 344, 355, 360, 375, 382, 390, 392, 398, 400 and 422 of a heavy chain of the antibody, and wherein the positions are numbered according to the EU system.
  • a modified antibody or antibody fragment comprises a substitution of one or more amino acids with cysteine on its constant region selected from positions 107, 108, 109, 114, 129, 142, 143, 145, 152, 154, 156, 159, 161, 165, 168, 169, 170, 182, 183, 197, 199, and 203 of a light chain of the antibody or antibody fragment, wherein the positions are numbered according to the EU system, and wherein the light chain is a human kappa light chain.
  • a modified antibody or antibody fragment thereof comprises a combination of substitution of two or more amino acids with cysteine on its constant regions wherein the combinations comprise substitutions at positions 375 of an antibody heavy chain, position 152 of an antibody heavy chain, position 360 of an antibody heavy chain, or position 107 of an antibody light chain and wherein the positions are numbered according to the EU system.
  • a modified antibody or antibody fragment thereof comprises a substitution of one amino acid with cysteine on its constant regions wherein the substitution is position 375 of an antibody heavy chain, position 152 of an antibody heavy 19 ME150999036v.1 132043-01120 chain, position 360 of an antibody heavy chain, position 107 of an antibody light chain, position 165 of an antibody light chain or position 159 of an antibody light chain and wherein the positions are numbered according to the EU system, and wherein the light chain is a kappa chain.
  • a modified antibody or antibody fragment thereof comprises a combination of substitution of two amino acids with cysteine on its constant regions wherein the combinations comprise substitutions at positions 375 of an antibody heavy chain and position 152 of an antibody heavy chain, wherein the positions are numbered according to the EU system.
  • a modified antibody or antibody fragment thereof comprises a substitution of one amino acid with cysteine at position 360 of an antibody heavy chain, wherein the positions are numbered according to the EU system.
  • a modified antibody or antibody fragment thereof comprises a substitution of one amino acid with cysteine at position 107 of an antibody light chain and wherein the positions are numbered according to the EU system, and wherein the light chain is a kappa chain.
  • antibody fragment or “antigen-binding fragment” or “functional antibody fragment,” as used herein, refers to at least one portion of an antibody that retains the ability to specifically interact with (e.g., by binding, steric hinderance, stabilizing/destabilizing, spatial distribution) an epitope of an antigen (e.g., CD74).
  • Antigen-binding fragments may also retain the ability to internalize into an antigen-expressing cell. In some embodiments, antigen-binding fragments also retain immune effector activity.
  • the terms antibody, antibody fragment, antigen- binding fragment, and the like, are intended to embrace the use of binding domains from antibodies in the context of larger macromolecules such as ADCs.
  • fragments of a full-length antibody can perform the antigen binding function of a full-length antibody.
  • antibody fragments include, but are not limited to, Fab, Fab’, F(ab’)2, Fv fragments, scFv antibody fragments, disulfide-linked Fvs (sdFv), a Fd fragment consisting of the VH and CH1 domains, linear antibodies, single domain antibodies such as sdAb (either VL or VH), camelid VHH domains, multi-specific antibodies formed from antibody fragments such as a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region, and an isolated CDR or other epitope binding fragments of an antibody.
  • An antigen-binding fragment can also be incorporated into single domain antibodies, maxibodies, minibodies, nanobodies, intrabodies, diabodies, triabodies, tetrabodies, bispecific or multi-specific antibody constructs, ADCs, v-NAR and bis-scFv (see, e.g., Holliger and Hudson (2005) Nat Biotechnol. 23(9):1126-36).
  • Antigen-binding fragments can also be grafted into scaffolds based on polypeptides such as a fibronectin type III (Fn3) (see US Patent No. 6,703,199, which describes fibronectin polypeptide minibodies).
  • scFv refers to a fusion protein comprising at least one antigen-binding fragment comprising a variable region of a light chain and at least one antigen-binding fragment comprising a variable region of a heavy chain, wherein the light and heavy chain variable regions are contiguously linked, e.g., via a synthetic linker, e.g., a short flexible polypeptide linker, and capable of being expressed as a single chain polypeptide, and wherein 20 ME150999036v.1 132043-01120 the scFv retains the specificity of the intact antibody from which it is derived.
  • a synthetic linker e.g., a short flexible polypeptide linker
  • an scFv may have the VL and VH variable regions in either order, e.g., with respect to the N-terminal and C-terminal ends of the polypeptide, the scFv may comprise VL-linker-VH or may comprise VH- linker-VL.
  • Antigen-binding fragments are obtained using conventional techniques known to those of skill in the art, and the binding fragments are screened for utility (e.g., binding affinity, internalization) in the same manner as are intact antibodies.
  • Antigen-binding fragments for example, may be prepared by cleavage of the intact protein, e.g., by protease or chemical cleavage.
  • CDR complementarity determining region
  • HCDR1, HCDR2, and HCDR3 three CDRs in each heavy chain variable region
  • LCDR1, LCDR2, and LCDR3 three CDRs in each light chain variable region
  • the precise amino acid sequence boundaries of a given CDR can be determined using any of a number of well-known schemes, including those described by Kabat et al. (1991) “Sequences of Proteins of Immunological Interest,” 5th Ed.
  • the CDRs correspond to the amino acid residues that are defined as part of the Kabat CDR, together with the amino acid residues that are defined as part of the Chothia CDR.
  • the CDRs defined according to the “Chothia” number scheme are also sometimes referred to as “hypervariable loops.”
  • the CDR amino acid residues in the heavy chain variable domain (VH) are numbered 31-35 (HCDR1) (e.g., insertion(s) after position 35), 50-65 (HCDR2), and 95-102 (HCDR3); and the CDR amino acid residues in the light chain variable domain (VL) are numbered 24-34 (LCDR1) (e.g., insertion(s) after position 27), 50-56 (LCDR2), and 89-97 (LCDR3).
  • the CDR amino acids in the VH are numbered 26-32 (HCDR1) (e.g., insertion(s) after position 31), 52-56 (HCDR2), and 95-102 (HCDR3); and the amino acid residues in VL are numbered 26-32 (LCDR1) (e.g., insertion(s) after position 30), 50-52 (LCDR2), and 91-96 (LCDR3).
  • the CDRs comprise or consist of, e.g., amino acid residues 26-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3) in human VH and amino acid residues 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3) in human VL.
  • the CDR amino acid residues in the VH are numbered approximately 26-35 (CDR1), 51-57 (CDR2) and 93-102 (CDR3), and the CDR amino acid residues in the VL are numbered approximately 27-32 (CDR1), 50- 52 (CDR2), and 89-97 (CDR3).
  • the CDR regions of an antibody may be determined using the program IMGT/DomainGap Align.
  • the term "monoclonal antibody,” as used herein, refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic epitope. In contrast, conventional (polyclonal) antibody preparations typically include a multitude of antibodies directed against (or specific for) different epitopes. The modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method.
  • the monoclonal antibodies to be used in accordance with the present disclosure may be made by the hybridoma method first described by Kohler et al. (1975) Nature 256:495, or may be made by recombinant DNA methods (see, e.g., US Patent No. 4,816,567).
  • Monoclonal antibodies may also be isolated from phage antibody libraries using the techniques described in Clackson et al. (1991) Nature 352:624-8, and Marks et al. (1991) J Mol Biol. 222:581-97, for example.
  • the term also includes preparations of antibody molecules of single molecular composition.
  • a monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope.
  • the monoclonal antibodies described herein can be non-human, human, or humanized.
  • the term specifically includes "chimeric" antibodies, in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they specifically bind the target antigen and/or exhibit the desired biological activity.
  • the term “human antibody,” as used herein, refers an antibody produced by a human or an antibody having an amino acid sequence of an antibody produced by a human.
  • the term includes antibodies having variable regions in which both the framework and CDR regions are derived from sequences of human origin. Furthermore, if the antibody contains a constant region, the constant region is also derived from such human sequences, e.g., human germline sequences, or mutated versions of human germline sequences or antibody containing consensus framework sequences derived from human framework sequences analysis, for example, as described in Knappik et al. ((2000) J Mol Biol. 296(1):57-86).
  • immunoglobulin variable domains e.g., CDRs
  • CDRs may be defined using well known numbering schemes, e.g., the Kabat numbering scheme, the Chothia numbering scheme, or a combination of Kabat and Chothia, and/or ImMunoGenTics (IMGT) numbering.
  • the human antibodies of the invention may include amino acid residues not encoded by human sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo, or a conservative substitution to promote stability or manufacturing).
  • human antibody is not intended to include 22 ME150999036v.1 132043-01120 antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.
  • recombinant human antibody refers to a human antibody that is prepared, expressed, created, or isolated by recombinant means, such as antibodies isolated from an animal (e.g., a mouse) that is transgenic or transchromosomal for human immunoglobulin genes or a hybridoma prepared therefrom, antibodies isolated from a host cell transformed to express the human antibody, e.g., from a transfectoma, antibodies isolated from a recombinant, combinatorial human antibody library, and antibodies prepared, expressed, created or isolated by any other means that involve splicing of all or a portion of a human immunoglobulin gene, sequences to other DNA sequences.
  • recombinant means such as antibodies isolated from an animal (e.g., a mouse) that is transgenic or transchromosomal for human immunoglobulin genes or a hybridoma prepared therefrom, antibodies isolated from a host cell transformed to express the human antibody, e.g., from a transfectoma, antibodies isolated from a
  • Such recombinant human antibodies have variable regions in which the framework and CDR regions are derived from human germline immunoglobulin sequences.
  • such recombinant human antibodies can be subjected to in vitro mutagenesis (or, when an animal transgenic for human Ig sequences is used, in vivo somatic mutagenesis) and thus the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that, while derived from and related to human germline VH and VL sequences, may not naturally exist within the human antibody germline repertoire in vivo.
  • chimeric antibody refers to antibodies wherein the amino acid sequence of the immunoglobulin molecule is derived from two or more species.
  • the variable regions of both heavy and light chains correspond to the variable regions of antibodies derived from one species with the desired specificity, affinity, and activity while the constant regions are homologous to antibodies derived from another species (e.g., human) to minimize an immune response in the latter species.
  • humanized antibody refers to forms of antibodies that contain sequences from non-human (e.g., murine) antibodies as well as human antibodies. Such antibodies are a type of chimeric antibody which contain minimal sequence derived from non-human immunoglobulin.
  • the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the framework (FR) regions are those of a human immunoglobulin sequence.
  • the humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.
  • Fc immunoglobulin constant region
  • the humanized antibody can be further modified by the substitution of residues, either in the Fv framework region and/or within the replaced non-human residues to refine and optimize antibody specificity, affinity, and/or activity.
  • Internalizing refers to an antibody or antigen-binding fragment that is capable of being taken through the cell’s lipid bilayer membrane to an internal compartment (i.e., “internalized”) upon binding to the cell, preferably into a degradative compartment in the cell.
  • an internalizing anti-CD74 antibody is one 23 ME150999036v.1 132043-01120 that is capable of being taken into the cell after binding to CD74 on the cell membrane.
  • the antibody or antigen-binding fragment used in the ADCs disclosed herein targets a cell surface antigen (e.g., CD74) and is an internalizing antibody or internalizing antigen-binding fragment (i.e., the ADC transfers through the cellular membrane after antigen binding).
  • the internalizing antibody or antigen-binding fragment binds a receptor on the cell surface.
  • An internalizing antibody or internalizing antigen-binding fragment that targets a receptor on the cell membrane may induce receptor-mediated endocytosis.
  • the internalizing antibody or internalizing antigen-binding fragment is taken into the cell via receptor- mediated endocytosis.
  • Non-internalizing as used herein in reference to an antibody or antigen-binding fragment refers to an antibody or antigen-binding fragment that remains at the cell surface upon binding to the cell.
  • the antibody or antigen-binding fragment used in the ADCs disclosed herein targets a cell surface antigen and is a non-internalizing antibody or non-internalizing antigen- binding fragment (i.e., the ADC remains at the cell surface and does not transfer through the cellular membrane after antigen binding).
  • cluster of differentiation 74 refers to any native form of human CD74 (also known as HLA class II histocompatibility antigen gamma chain or HLA-DR antigens-associated invariant chain).
  • the term encompasses full-length human CD74 (e.g., NCBI Reference Sequence: NP_001020330.1; SEQ ID NO:22), as well as any form of human CD74 that may result from cellular processing.
  • CD74 can be isolated from human, or may be produced recombinantly or by synthetic methods.
  • anti-CD74 antibody or “antibody that binds to CD74,” as used herein, refers to any form of antibody or antigen-binding fragment thereof that binds, e.g., specifically binds, to CD74.
  • the term encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and biologically functional antigen-binding fragments so long as they bind, e.g., specifically bind, to CD74.
  • WO2020/236817 provides and is incorporated herein by reference for exemplary CD74-binding sequences, including exemplary anti-CD74 antibody sequences.
  • the anti-CD74 antibody used in the ADCs disclosed herein is an internalizing antibody or internalizing antigen-binding fragment.
  • VHmil x VK1aNQ (WO2020/236817) is an exemplary anti-CD74 antibody.
  • binding specificity refers to the ability of an individual antibody or antigen binding fragment to preferentially react with one antigenic determinant over a different antigenic determinant. The degree of specificity indicates the extent to which an antibody or fragment preferentially binds to one antigenic determinant over a different antigenic determinant. Also, as used 24 ME150999036v.1 132043-01120 herein, the terms “specific,” “specifically binds,” and “binds specifically” refer to a binding reaction between an antibody or antigen-binding fragment (e.g., an anti-CD74 antibody) and a target antigen (e.g., CD74) in a heterogeneous population of proteins and other biologics.
  • an antibody or antigen-binding fragment e.g., an anti-CD74 antibody
  • a target antigen e.g., CD74
  • Antibodies can be tested for specificity of binding by comparing binding to an appropriate antigen to binding to an irrelevant antigen or antigen mixture under a given set of conditions. If the antibody binds to the appropriate antigen with at least 2, 5, 7, 10 or more times more affinity than to the irrelevant antigen or antigen mixture, then it is considered to be specific.
  • a “specific antibody” or a “target-specific antibody” is one that only binds the target antigen (e.g., CD74), but does not bind (or exhibits minimal binding) to other antigens.
  • an antibody or antigen-binding fragment that specifically binds a target antigen has a KD of less than 1x10 -6 M, less than 1x10 -7 M, less than 1x10 -8 M, less than 1x10 -9 M, less than 1x10 -10 M, less than 1x10 -11 M, less than 1x10 -12 M, or less than 1x10 -13 M.
  • the K D is 1 pM to 500 pM. In some embodiments, the K D is between 500 pM to 1 ⁇ M, 1 ⁇ M to 100 nM, or 100 mM to 10 nM.
  • the term “affinity,” as used herein, refers to the strength of interaction between antibody and antigen at single antigenic sites. Without being bound by theory, within each antigen binding site, the variable region of the antibody “arm” interacts through weak non-covalent forces with the antigen at numerous sites; the more interactions, typically the stronger the affinity.
  • the binding affinity of an antibody is the sum of the attractive and repulsive forces operating between the antigenic determinant and the binding site of the antibody.
  • the term “kon” or “ka” refers to the on-rate constant for association of an antibody to the antigen to form the antibody/antigen complex. The rate can be determined using standard assays, such as a surface plasmon resonance, biolayer inferometry, or ELISA assay.
  • the term "koff” or “kd” refers to the off-rate constant for dissociation of an antibody from the antibody/antigen complex. The rate can be determined using standard assays, such as a surface plasmon resonance, biolayer inferometry, or ELISA assay.
  • KD refers to the equilibrium dissociation constant of a particular antibody-antigen interaction. K D is calculated by k a /k d . The rate can be determined using standard assays, such as a surface plasmon resonance, biolayer inferometry, or ELISA assay.
  • the term “epitope” refers to the portion of an antigen capable of being recognized and specifically bound by an antibody (or antigen-binding fragment).
  • Epitope determinants generally consist of chemically active surface groupings of molecules such as amino acids or carbohydrate or sugar side chains and can have specific three-dimensional structural characteristics, as well as specific charge characteristics.
  • epitopes can be formed from contiguous amino acids or noncontiguous amino acids juxtaposed by tertiary folding of the polypeptide.
  • An epitope may be “linear” or “conformational.” Conformational and linear epitopes are distinguished in that the binding to the former but not the latter is lost in the presence of denaturing solvents.
  • the epitope bound by an antibody may be identified using any epitope 25 ME150999036v.1 132043-01120 mapping technique known in the art, including X-ray crystallography for epitope identification by direct visualization of the antigen-antibody complex, as well as monitoring the binding of the antibody to fragments or mutated variations of the antigen, or monitoring solvent accessibility of different parts of the antibody and the antigen.
  • Exemplary strategies used to map antibody epitopes include, but are not limited to, array-based oligo-peptide scanning, limited proteolysis, site-directed mutagenesis, high-throughput mutagenesis mapping, hydrogen-deuterium exchange, and mass spectrometry (see, e.g., Gershoni et al.
  • competitive binding is identified when a test antibody or binding protein reduces binding of a reference antibody or binding protein to a target antigen such as CD74 (e.g., a binding protein comprising CDRs and/or variable domains selected from those identified in Tables A2 and A3), by at least about 50% in the cross- blocking assay (e.g., 50%, 60%, 70%, 80%, 90%, 95%, 99%, 99.5%, or more, or any percentage in between), and/or vice versa.
  • a target antigen such as CD74
  • competitive binding can be due to shared or similar (e.g., partially overlapping) epitopes, or due to steric hindrance where antibodies or binding proteins bind at nearby epitopes (see, e.g., Tzartos, Methods in Molecular Biology (Morris, ed. (1998) vol. 66, pp. 55-66)).
  • competitive binding can be used to sort groups of binding proteins that share similar epitopes. For example, binding proteins that compete for binding can be “binned” as a group of binding proteins that have overlapping or nearby epitopes, while those that do not compete are placed in a separate group of binding proteins that do not have overlapping or nearby epitopes.
  • peptide As used herein, the terms “peptide,” “polypeptide,” and “protein” are used interchangeably to refer to a polymer of amino acid residues.
  • the terms encompass amino acid polymers comprising two or more amino acids joined to each other by peptide bonds, amino acid polymers in which one or more amino acid residues is an artificial chemical mimetic of a corresponding naturally-occurring amino acid, as well as naturally-occurring amino acid polymers and non-naturally-occurring amino acid polymers.
  • the terms include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others.
  • a "recombinant” protein refers to a protein (e.g., an antibody) made using recombinant techniques, e.g., through the expression of a recombinant nucleic acid. 26 ME150999036v.1 132043-01120 [87] An "isolated" protein refers to a protein unaccompanied by at least some of the material with which it is normally associated in its natural state.
  • an "isolated antibody,” as used herein, is an antibody that has been identified and separated from one or more (e.g., the majority) of the components (by weight) of its source environment, e.g., from the components of a hybridoma cell culture or a different cell culture that was used for its production.
  • the separation is performed such that it sufficiently removes components that may otherwise interfere with the suitability of the antibody for the desired applications (e.g., for therapeutic use).
  • Methods for preparing isolated antibodies include, without limitation, protein A chromatography, anion exchange chromatography, cation exchange chromatography, virus retentive filtration, and ultrafiltration.
  • variant refers to a nucleic acid sequence or an amino acid sequence that differs from a reference nucleic acid sequence or amino acid sequence respectively, but retains one or more biological properties of the reference sequence.
  • a variant may contain one or more amino acid substitutions, deletions, and/or insertions (or corresponding substitution, deletion, and/or insertion of codons) with respect to a reference sequence. Changes in a nucleic acid variant may not alter the amino acid sequence of a peptide encoded by the reference nucleic acid sequence, or may result in amino acid substitutions, additions, deletions, fusions, and/or truncations.
  • a nucleic acid variant disclosed herein encodes an identical amino acid sequence to that encoded by the unmodified nucleic acid or encodes a modified amino acid sequence that retains one or more functional properties of the unmodified amino acid sequence.
  • a variant of a nucleic acid or peptide can be a naturally-occurring variant or a variant that is not known to occur naturally. Variants of nucleic acids and peptides may be made by mutagenesis techniques, by direct synthesis, or by other techniques known in the art. A variant does not necessarily require physical manipulation of the reference sequence.
  • a variant has high sequence identity (i.e., 60% nucleic acid or amino acid sequence identity or higher) as compared to a reference sequence.
  • a peptide variant encompasses polypeptides having amino acid 27 ME150999036v.1 132043-01120 substitutions, deletions, and/or insertions as long as the polypeptide has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% amino acid sequence identity with a reference sequence, or with a corresponding segment (e.g., a functional fragment) of a reference sequence, e.g., those variants that also retain one or more functions of the reference sequence.
  • a corresponding segment e.g., a functional fragment
  • a nucleic acid variant encompasses polynucleotides having amino acid substitutions, deletions, and/or insertions as long as the polynucleotide has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% nucleic acid sequence identity with a reference sequence, or with a corresponding segment (e.g., a functional fragment) of a reference sequence.
  • the term “conservatively modified variant” applies to both amino acid and nucleic acid sequences.
  • nucleic acid sequences conservatively modified variants refer to those nucleic acids which encode identical or essentially identical amino acid sequences. Because of the degeneracy of the genetic code, a large number of functionally identical nucleic acids encode any given protein. For instance, the codons GCA, GCC, GCG and GCU all encode the amino acid alanine. Thus, at every position where an alanine is specified by a codon, the codon can be altered to any of the corresponding codons described without altering the encoded polypeptide. Such nucleic acid variations are “silent variations,” which are one species of conservatively modified variations. Every nucleic acid sequence herein which encodes a polypeptide also describes every possible silent variation of the nucleic acid.
  • each codon in a nucleic acid can be modified to yield a functionally identical molecule. Accordingly, each silent variation of a nucleic acid that encodes a polypeptide is implicit in each described sequence.
  • conservatively modified variants include individual substitutions, deletions, or additions to a polypeptide sequence which result in the substitution of an amino acid with a chemically similar amino acid. Conservative substitutions providing functionally similar amino acids are well known in the art.
  • conservative sequence modifications refers to amino acid modifications that do not significantly affect or alter the binding characteristics of, e.g., an antibody or antigen-binding fragment containing the amino acid sequence. Such conservative modifications include amino acid substitutions, additions, and deletions. Modifications can be introduced into an antibody or antigen-binding fragment by standard techniques known in the art, such as, e.g., site- directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions are ones in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art.
  • amino acids with basic side chains e.g., lysine, arginine, histidine
  • acidic side chains 28 ME150999036v.1 132043-01120 e.g., aspartic acid, glutamic acid
  • uncharged polar side chains e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan
  • nonpolar side chains e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine
  • beta-branched side chains e.g., threonine, valine, isoleucine
  • aromatic side chains e.g., tyrosine, phenylalanine, tryptophan, histidine
  • one or more amino acid residues within an antibody can be replaced with other amino acid residues from the same side chain family and the altered antibody can be tested using the functional assays described herein.
  • the term “homologous” or “identity,” as used herein, refers to the subunit sequence identity between two polymeric molecules, e.g., between two nucleic acid molecules, such as, two DNA molecules or two RNA molecules, or between two polypeptide molecules. When a subunit position in both of the two molecules is occupied by the same monomeric subunit; e.g., if a position in each of two DNA molecules is occupied by adenine, then they are homologous or identical at that position.
  • the homology between two sequences is a direct function of the number of matching or homologous positions. For example, if half (e.g., five positions in a polymer ten subunits in length) of the positions in two sequences are matched or homologous, the two sequences are 50% homologous; if 90% of the positions (e.g., 9 of 10), are matched or homologous, the two sequences are 90% homologous.
  • Percentage of “sequence identity” can be determined by comparing two optimally aligned sequences over a comparison window, where the fragment of the amino acid sequence in the comparison window may comprise additions or deletions (e.g., gaps or overhangs) as compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences.
  • the percentage can be calculated by determining the number of positions at which the identical amino acid residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison, and multiplying the result by 100 to yield the percentage of sequence identity.
  • the output is the percent identity of the subject sequence with respect to the query sequence.
  • the percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences.
  • amino acid identity or homology between proteins disclosed herein and variants thereof, including variants of target antigens (such as CD74) and variants of antibody variable domains (including individual variant CDRs) is at least 80% to the sequences depicted herein, e.g., identities or homologies of at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, almost 100%, or 100%.
  • the comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm.
  • the percent identity between two amino acid sequences is determined using the Needleman and Wunsch ((1970) J Mol Biol. 48:444-53) algorithm which has been incorporated into the GAP program in the GCG software 29 ME150999036v.1 132043-01120 package, using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6.
  • the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package, using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6.
  • An exemplary set of parameters is a Blossum 62 scoring matrix with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5.
  • the percent identity between two amino acid or nucleotide sequences can also be determined using the algorithm of Meyers and Miller ((1989) CABIOS 4:11-17) which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.
  • agent is used herein to refer to a chemical compound, a mixture of chemical compounds, a biological macromolecule, an extract made from biological materials, or a combination of two or more thereof.
  • therapeutic agent or “drug” refers to an agent that is capable of modulating a biological process and/or has biological activity.
  • the BH3 mimetics and the ADCs comprising them, as described herein, are exemplary therapeutic agents.
  • chemotherapeutic agent or “anti-cancer agent” is used herein to refer to all agents that are effective in treating cancer (regardless of mechanism of action). Inhibition of metastasis or angiogenesis is frequently a property of a chemotherapeutic agent.
  • Chemotherapeutic agents include antibodies, biological molecules, and small molecules, and encompass the BH3 mimetics and ADCs comprising them, as described herein.
  • a chemotherapeutic agent may be a cytotoxic or cytostatic agent.
  • cytostatic agent refers to an agent that inhibits or suppresses cell growth and/or multiplication of cells.
  • cytotoxic agent refers to a substance that causes cell death primarily by interfering with a cell’s expression activity and/or functioning.
  • antigenoplastic payload or “antineoplastic compound” as used herein, refers to a compound or compounds that slow or inhibit the division of cancerous cells or that kill the cancerous cells.
  • Non-limiting examples of antineoplastic payloads include the drugs described herein.
  • the term “BH3 mimetic,” as used herein refers to an agent capable of disrupting the interaction between the proapoptotic and antiapoptotic members of the Bcl-2 family and are potent inducers of apoptosis.
  • Exemplary BH3 mimetics include inhibitors of Bcl-2, Bcl-xL, Bcl-w and Mcl- 1.
  • Mcl-1 myeloid cell leukemia 1
  • Mcl-1 refers to any native form of human Mcl-1, an anti-apoptotic member of the Bcl-2 protein family.
  • Mcl-1 encompasses full- length human Mcl-1 (e.g., UniProt Reference Sequence: Q07820), as well as any form of human Mcl- 1 that may result from cellular processing.
  • the term also encompasses functional variants or fragments of human Mcl-1, including but not limited to splice variants, allelic variants, and isoforms that retain one or more biologic functions of human Mcl-1 (i.e., variants and fragments are encompassed unless the context indicates that the term is used to refer to the wild-type protein only).
  • Mcl-1 can be isolated from human, or may be produced recombinantly or by synthetic methods.
  • inhibitor means to reduce a biological activity or process by a measurable amount, and can include but does not require complete prevention or inhibition.
  • “inhibition” means to reduce the expression and/or activity of a Bcl-2 family protein (e.g., Bcl-2, Bcl-xL, Mcl-1) and/or one or more upstream modulators or downstream targets thereof.
  • Mcl-1 inhibitor refers to an agent capable of reducing the expression and/or activity of Mcl-1 and/or one or more upstream modulators or downstream targets thereof.
  • Mcl-1 modulators are described in WO 2015/097123; WO 2016/207216; WO 2016/207217; WO 2016/207225; WO 2016/207226; WO 2017/125224; WO 2019/035899, WO 2019/035911, WO 2019/035914, WO 2019/035927, US 2019/0055264, WO 2016/033486, WO 2017/147410, WO 2018/183418, and WO 2017/182625, each of which are incorporated herein by reference as exemplary Mcl-1 modulators, including exemplary Mcl-1 inhibitors, that can be included as drug moieties in the disclosed ADCs.
  • Mcl-1 inhibitor when referring to an Mcl-1 inhibitor, or the like, means any such compound that retains essentially the same, similar, or enhanced biological function or activity as compared to the original compound but has an altered chemical or biological structure.
  • a “Mcl-1 inhibitor drug moiety”, “Mcl-1 inhibitor”, and the like refer to the component of an ADC or composition that provides the structure of an Mcl-1 inhibitor compound or a compound modified for attachment to an ADC that retains essentially the same, similar, or enhanced biological function or activity as compared to the original compound.
  • B-cell lymphoma-extra large refers to any native form of human Bcl-xL, an anti-apoptotic member of the Bcl-2 protein family.
  • the term encompasses full-length human Bcl-xL (e.g., UniProt Reference Sequence: Q07817-1), as well as any form of human Bcl-xL that may result from cellular processing.
  • Bcl-xL can be isolated from human, or may be produced recombinantly or by synthetic methods.
  • Bcl-xL inhibitor refers to an agent capable of reducing the expression and/or activity of Bcl-xL and/or one or more upstream modulators or downstream targets thereof.
  • Exemplary Bcl-xL modulators are described in WO2010/080503, WO2010/080478, WO2013/055897, WO2013/055895, WO2016/094509, WO2016/094517, WO2016/094505, WO 2021/018858, WO 2021/018857, Tao et al., ACS Medicinal Chemistry Letters (2014), 5(10), 1088-109, and Wang et al., ACS Medicinal Chemistry Letters (2020), 11(10), 1829 ⁇ 1836, each of which are incorporated herein by reference as exemplary Bcl-xL 31 ME150999036v.1 132043-01120 modulators, including exemplary Bcl-xL inhibitors, that can be included as drug moieties in the disclosed ADCs.
  • a “Bcl-xL inhibitor drug moiety”, “Bcl-xL inhibitor”, and the like refer to the component of an ADC or composition that provides the structure of a Bcl-xL inhibitor compound or a compound modified for attachment to an ADC that retains essentially the same, similar, or enhanced biological function or activity as compared to the original compound.
  • the term encompasses full-length human Bcl-2 (e.g., UniProt Reference Sequence: P10415), as well as any form of human Bcl-2 that may result from cellular processing.
  • the term also encompasses functional variants or fragments of human Bcl-2, including but not limited to splice variants, allelic variants, and isoforms that retain one or more biologic functions of human Mcl-1 (i.e., variants and fragments are encompassed unless the context indicates that the term is used to refer to the wild-type protein only).
  • Bcl-2 can be isolated from human, or may be produced recombinantly or by synthetic methods.
  • Bcl-2 inhibitor refers to an agent capable of reducing the expression and/or activity of Bcl-2 and/or one or more upstream modulators or downstream targets thereof.
  • Exemplary Bcl-2 modulators are described in WO 2013/110890, WO 2015/011400, WO 2015/011399, WO 2015/011397, WO 2015/011396, WO 2015/011164 and WO 2019081559, each of which are incorporated herein by reference as exemplary Bcl-2 modulators, including exemplary Bcl-2 inhibitors, that can be included as drug moieties in the disclosed ADCs.
  • a “Bcl-2 inhibitor drug moiety”, “Bcl-2 inhibitor”, and the like refer to the component of an ADC or composition that provides the structure of a Bcl-2 inhibitor compound or a compound modified for attachment to an ADC that retains essentially the same, similar, or enhanced biological function or activity as compared to the original compound.
  • the term “cancer,” as used herein, refers to the presence of cells possessing characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and/or certain morphological features.
  • cancer cells can be in the form of a tumor or mass, but such cells may exist alone within a subject, or may circulate in the blood stream as independent cells, such as leukemic or lymphoma cells.
  • the term "cancer” includes all types of cancers and cancer metastases, including hematological cancers, solid tumors, sarcomas, carcinomas and other solid and non-solid tumor cancers.
  • Hematological cancers may include B-cell malignancies, cancers of the blood (leukemias), cancers of plasma cells (myelomas, e.g., multiple myeloma), or cancers of the lymph nodes (lymphomas).
  • Exemplary B-cell malignancies include chronic lymphocytic leukemia (CLL), follicular lymphoma, mantle cell lymphoma, and diffuse large B-cell lymphoma.
  • Leukemias may include acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), 32 ME150999036v.1 132043-01120 chronic myelomonocytic leukemia (CMML), acute monocytic leukemia (AMoL), etc.
  • ALL acute lymphoblastic leukemia
  • AML acute myeloid leukemia
  • CLL chronic lymphocytic leukemia
  • CML chronic myelogenous leukemia
  • CMML chronic myelomonocytic leukemia
  • AoL acute monocytic leukemia
  • Lymphomas may include Hodgkin's lymphoma, non-Hodgkin's lymphoma, etc.
  • Other hematologic cancers may include myelodysplasia syndrome (MDS).
  • Solid tumors may include carcinomas such as adenocarcinoma, e.g., breast cancer, pancreatic cancer, prostate cancer, colon or colorectal cancer, lung cancer, gastric cancer, cervical cancer, endometrial cancer, ovarian cancer, cholangiocarcinoma, glioma, melanoma, etc.
  • the cancer is a breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular cancer, lymphoblastic leukemia including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T- cell or B-cell origin, melanoma, myelogenous leukemia, myeloma, oral cancer, ovarian cancer, non- small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, stomach cancer, colon cancer, or head and neck cancer.
  • the cancer is acute myeloid leukemia, multiple myeloma, or B-cell lymphoma.
  • tumor refers to any mass of tissue that results from excessive cell growth or proliferation, either benign or malignant, including precancerous lesions.
  • the tumor is a breast cancer, gastric cancer, bladder cancer, brain cancer, cervical cancer, colorectal cancer, esophageal cancer, hepatocellular cancer, melanoma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, pancreatic cancer, stomach cancer, colon cancer, head and neck cancer, or spleen cancer.
  • tumor cell and “cancer cell” may be used interchangeably herein and refer to individual cells or the total population of cells derived from a tumor or cancer, including both non- tumorigenic cells and cancer stem cells.
  • tumor cell and “cancer cell” will be modified by the term “non-tumorigenic” when referring solely to those cells lacking the capacity to renew and differentiate to distinguish those cells from cancer stem cells.
  • target-negative refers to the absence of target antigen expression by a cell or tissue.
  • target-positive target antigen-positive
  • antigen-positive refers to the presence of target antigen expression.
  • Non-human animals include all vertebrates (e.g., mammals and non-mammals) such as any mammal.
  • mammals include humans, chimpanzees, apes, monkeys, cattle, horses, sheep, goats, swine, rabbits, dogs, cats, rats, mice, and guinea pigs.
  • non-mammals include birds and fish.
  • the subject is a human. 33 ME150999036v.1 132043-01120
  • a subject in need of treatment refers to a subject that would benefit biologically, medically, or in quality of life from a treatment (e.g., a treatment with any one or more of the exemplary ADC compounds described herein).
  • the term “treat,” “treating,” or “treatment” refers to any improvement of any consequence of disease, disorder, or condition, such as prolonged survival, less morbidity, and/or a lessening of side effects which result from an alternative therapeutic modality.
  • treatment comprises delaying or ameliorating a disease, disorder, or condition (i.e., slowing or arresting or reducing the development of a disease or at least one of the clinical symptoms thereof).
  • treatment comprises delaying, alleviating, or ameliorating at least one physical parameter of a disease, disorder, or condition, including those which may not be discernible by the patient.
  • treatment comprises modulating a disease, disorder, or condition, either physically (e.g., stabilization of a discernible symptom), physiologically (e.g., stabilization of a physical parameter), or both.
  • treatment comprises administration of a described ADC compound or composition to a subject, e.g., a patient, to obtain a treatment benefit enumerated herein.
  • the treatment can be to cure, heal, alleviate, delay, prevent, relieve, alter, remedy, ameliorate, palliate, improve, or affect a disease, disorder, or condition (e.g., a cancer), the symptoms of a disease, disorder, or condition (e.g., a cancer), or a predisposition toward a disease, disorder, or condition (e.g., a cancer).
  • a composition disclosed herein in addition to treating a subject having a disease, disorder, or condition, can also be provided prophylactically to prevent or reduce the likelihood of developing that disease, disorder, or condition.
  • the term “prevent”, “preventing,” or “prevention” of a disease, disorder, or condition refers to the prophylactic treatment of the disease, disorder, or condition; or delaying the onset or progression of the disease, disorder, or condition.
  • a "pharmaceutical composition” refers to a preparation of a composition, e.g., an ADC compound or composition, in addition to at least one other (and optionally more than one other) component suitable for administration to a subject, such as a pharmaceutically acceptable carrier, stabilizer, diluent, dispersing agent, suspending agent, thickening agent, and/or excipient.
  • a pharmaceutically acceptable carrier such as a pharmaceutically acceptable carrier, stabilizer, diluent, dispersing agent, suspending agent, thickening agent, and/or excipient.
  • the pharmaceutical compositions provided herein are in such form as to permit administration and subsequently provide the intended biological activity of the active ingredient(s) and/or to achieve a therapeutic effect.
  • the pharmaceutical compositions provided herein preferably contain no additional components which are unacceptably toxic to a subject to which the formulation would be administered.
  • Pharmaceutically acceptable carriers may enhance or stabilize the composition or can 34 ME150999036v.1 132043-01120 be used to facilitate preparation of the composition.
  • Pharmaceutically acceptable carriers can include solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial agents, antifungal agents), isotonic agents, absorption delaying agents, salts, preservatives, drug stabilizers, binders, excipients, disintegration agents, lubricants, sweetening agents, flavoring agents, dyes, and the like and combinations thereof, as would be known to those skilled in the art (see, for example, Remington's Pharmaceutical Sciences, 18th Ed. Mack Printing Company, 1990, pp.1289- 1329). Except insofar as any conventional carrier is incompatible with the active ingredient, its use in the therapeutic or pharmaceutical compositions is contemplated.
  • preservatives e.g., antibacterial agents, antifungal agents
  • isotonic agents e.g., absorption delaying agents, salts, preservatives, drug stabilizers, binders, excipients, disintegration agents, lubricants, sweetening
  • the carrier may be selected to minimize adverse side effects in the subject, and/or to minimize degradation of the active ingredient(s).
  • An adjuvant may also be included in any of these formulations.
  • the term "excipient” refers to an inert substance added to a pharmaceutical composition to further facilitate administration of an active ingredient.
  • Formulations for parenteral administration can, for example, contain excipients such as sterile water or saline, polyalkylene glycols such as polyethylene glycol, vegetable oils, or hydrogenated napthalenes.
  • excipients include, but are not limited to, calcium bicarbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, ethylene-vinyl acetate co-polymer particles, and surfactants, including, for example, polysorbate 20.
  • pharmaceutically acceptable salt refers to a salt which does not abrogate the biological activity and properties of the compounds of the invention, and does not cause significant irritation to a subject to which it is administered.
  • salts include, but are not limited to: (a) acid addition salts formed with inorganic acids, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid and the like; and salts formed with organic acids, for example, acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, polygalacturonic acid, and the like; and (b) salts formed from elemental anions such as chlorine, bromine, and iodine.
  • inorganic acids for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phospho
  • the antibody-drug conjugates (ADCs), linkers, payloads and linker-payloads described herein can contain a monovalent anionic counterion M1-. Any suitable anionic counterion can be used.
  • the monovalent anionic counterion is a pharmaceutically acceptable monovalent anionic counterion.
  • the monovalent anionic counterion M1- can be selected from bromide, chloride, iodide, acetate, trifluoroacetate, benzoate, mesylate, tosylate, triflate, formate, or the like. In some embodiments, the monovalent anionic counterion M1- is trifluoroacetate or formate.
  • the term “therapeutically effective amount” or “therapeutically effective dose,” refers to an amount of a compound described herein, e.g., an ADC compound or composition described herein, to effect the desired therapeutic result (i.e., reduction or inhibition of an enzyme or a protein activity, amelioration of symptoms, alleviation of symptoms or conditions, delay of disease progression, a reduction in tumor size, inhibition of tumor growth, prevention of metastasis).
  • a therapeutically effective amount does not induce or cause undesirable side effects.
  • a therapeutically effective amount induces or causes side effects but only those that are acceptable by a treating clinician in view of a patient’s condition.
  • a therapeutically effective amount is effective for detectable killing, reduction, and/or inhibition of the growth or spread of cancer cells, the size or number of tumors, and/or other measure of the level, stage, progression and/or severity of a cancer.
  • the term also applies to a dose that will induce a particular response in target cells, e.g., a reduction, slowing, or inhibition of cell growth.
  • a therapeutically effective amount can be determined by first administering a low dose, and then incrementally increasing that dose until the desired effect is achieved.
  • a therapeutically effective amount can also vary depending upon the intended application (in vitro or in vivo), or the subject and disease condition being treated, e.g., the weight and age of the subject, the severity of the disease condition, the manner of administration and the like, which can readily be determined by one of ordinary skill in the art.
  • the specific amount may vary depending on, for example, the particular pharmaceutical composition, the subject and their age and existing health conditions or risk for health conditions, the dosing regimen to be followed, the severity of the disease, whether it is administered in combination with other agents, timing of administration, the tissue to which it is administered, and the physical delivery system in which it is carried.
  • a therapeutically effective amount of an ADC may reduce the number of cancer cells, reduce tumor size, inhibit (e.g., slow or stop) tumor metastasis, inhibit (e.g., slow or stop) tumor growth, and/or relieve one or more symptoms.
  • the term “prophylactically effective amount” or “prophylactically effective dose,” refers to an amount of a compound disclosed herein, e.g., an ADC compound or composition described herein, that is effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease, the prophylactically effective amount will be less than the therapeutically effective amount.
  • a prophylactically effective amount can prevent the onset of disease symptoms, including symptoms associated with a cancer.
  • the term “p” or “drug loading” or “drug:antibody ratio” or “drug-to-antibody ratio” or “DAR” refers to the number of drug moieties per antibody or antigen-binding fragment, i.e., drug loading, or the number of antineoplastic payloads per antibody or antigen-binding fragment (Ab) in ADCs of Formula (I).
  • one dual linker attaches two antineoplastic compounds (e.g.
  • p is 2 if the antibody or antigen-binding fragment only links with one dual linker having two antineoplastic compounds (e.g. two BH3 mimetic drug moieties) attached thereto.
  • “average p” refers to the average number of antineoplastic compounds (e.g. two BH3 mimetic drug moieties) per antibody or antigen- binding fragment, also referred to as “average drug loading.” 1.
  • the antibody-drug conjugate (ADC) compounds of the present disclosure include those with anti-cancer activity.
  • the ADC compounds include an antibody or antigen-binding fragment conjugated (i.e., covalently attached by a dual linker) to two antineoplastic compounds as described herein.
  • the antineoplastic compound when not conjugated to an antibody or antigen-binding fragment is capable of reducing the expression and/or activity of a Bcl-2 family protein (e.g., Mcl-1, Bcl-2 and/or Bcl-xL) and/or one or more upstream modulators or downstream targets thereof.
  • a Bcl-2 family protein e.g., Mcl-1, Bcl-2 and/or Bcl-xL
  • the ADCs disclosed herein may provide potent anti-cancer agents.
  • the ADC may provide improved activity, better cytotoxic specificity, and/or reduced off-target killing as compared to the antineoplastic compound when administered alone.
  • the components of the ADC are selected to (i) retain one or more therapeutic properties exhibited by the antibody and antineoplastic compounds in isolation, (ii) maintain the specific binding properties of the antibody or antigen-binding fragment; (iii) optimize drug loading and drug-to-antibody ratios; (iv) allow delivery, e.g., intracellular delivery, of the antineoplastic compound via stable attachment to the antibody or antigen-binding fragment; (v) retain ADC stability as an intact conjugate until transport or delivery to a target site; (vi) minimize aggregation of the ADC prior to or after administration; (vii) allow for the therapeutic effect, e.g., cytotoxic effect, of the antineoplastic compound after cleavage or other release mechanism in the cellular environment; (viii) exhibit in vivo anti-cancer treatment efficacy comparable to or superior to that of the antibody and antineoplastic compounds in isolation; (ix) minimize off-target killing by the antineoplastic compound; and/or (
  • the ADC compounds of the present disclosure may selectively deliver an effective dose of a cytotoxic or cytostatic agent to cancer cells or to tumor tissue.
  • the cytotoxic and/or cytostatic activity of the ADC is dependent on target antigen expression in a cell.
  • the disclosed ADCs are particularly effective at killing cancer cells expressing a target 37 ME150999036v.1 132043-01120 antigen while minimizing off-target killing.
  • the disclosed ADCs do not exhibit a cytotoxic and/or cytostatic effect on cancer cells that do not express a target antigen.
  • ADC compounds comprising an antibody or antigen- binding fragment thereof (Ab) covalently linked to two antineoplastic payloads, such as a BH3 mimetic (D 1 and D 2 ) through a dual linker (L).
  • the antibody or antigen-binding fragment thereof (Ab) targets a cancer cell.
  • the antibody or antigen-binding fragment is able to bind to a tumor-associated antigen (e.g., CD74), e.g., with high specificity and high affinity.
  • the antibody or antigen-binding fragment is internalized into a target cell upon binding, e.g., into a degradative compartment in the cell.
  • the ADCs internalize upon binding to a target cell, undergo degradation, and release the antineoplastic payload to kill cancer cells.
  • the antineoplastic payloads such as BH3 mimetics, may be released from the antibody and/or the linker moiety of the ADC by enzymatic action, hydrolysis, oxidation, or any other mechanism.
  • the antibody or antigen-binding fragment (Ab) of Formula (I) includes within its scope an anti-CD74 antibody or antigen-binding fragment that specifically binds to a target antigen on a cell. In some embodiment, the antibody or antigen-binding fragment (Ab) of Formula (I) includes within its scope an anti-CD74 antibody or antigen-binding fragment that specifically binds to a target antigen on a cancer cell (e.g., CD74).
  • the antibody or antigen-binding fragment may bind to a target antigen with a dissociation constant (KD) of ⁇ 1 mM, ⁇ 100 nM or ⁇ 10 nM, or any amount in between, as measured by, e.g., BIAcore ® analysis.
  • KD dissociation constant
  • the KD is 1 pM to 500 pM.
  • the KD is between 500 pM to 1 ⁇ M, 1 ⁇ M to 100 nM, or 100 mM to 10 nM.
  • the antibody or antigen-binding fragment is a four-chain anti-CD74 antibody (also referred to as an immunoglobulin or a full-length or intact antibody), comprising two heavy chains and two light chains.
  • the antibody or antigen-binding fragment is an antigen-binding fragment of an immunoglobulin.
  • the antibody or antigen- binding fragment is an antigen-binding fragment of an immunoglobulin that retains the ability to bind a target cancer antigen and/or provide at least one function of the immunoglobulin.
  • the antibody or antigen-binding fragment is an internalizing anti-CD74 antibody or internalizing antigen-binding fragment thereof.
  • the internalizing anti-CD74 antibody or internalizing antigen-binding fragment thereof binds to a target cancer antigen expressed on the surface of a cell and enters the cell upon binding.
  • the antineoplastic payload of the ADC is released from the anti-CD74 antibody or antigen-binding fragment of the ADC after the ADC enters and is present in a cell expressing the target cancer antigen (i.e., after the ADC has been internalized), e.g., by cleavage, by degradation of the antibody or antigen-binding fragment, or by any other suitable release mechanism.
  • the anti-CD74 antibodies or antigen-binding fragments comprise mutations that mediate reduced or no antibody-dependent cellular cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC). In some embodiments, these mutations are known as Fc Silencing, Fc Silent, or Fc Silenced mutations.
  • ADCC antibody-dependent cellular cytotoxicity
  • CDC complement-dependent cytotoxicity
  • amino acid residues L234 and L235 of the IgG1 constant region are substituted to A234 and A235 (also known as “LALA”).
  • amino acid residue N297 of the IgG1 constant region is substituted to A297 (also known as “N297A”).
  • amino acid residues D265 and P329 of the IgG1 constant region are substituted to A265 and A329 (also known as “DAPA”).
  • Other antibody Fc silencing mutations may also be used.
  • the Fc silencing mutations are used in combination, for example D265A, N297A and P329A (also known as “DANAPA”).
  • D265A, N297A and P329A also known as “DANAPA”.
  • E152C, S375C according to EU numbering of the antibody heavy chain to facilitate conjugation to linker-drug moieties
  • CysMab the 40 ME150999036v.1 132043-01120 antibody or antigen-binding fragment has been modified with Fc silencing mutations D265A, N297A and P329A of the IgG1 constant region according to EU numbering
  • DANAPA is added to the antibody name
  • DAPA is added to the antibody name.
  • Amino acid sequences of exemplary antibodies or antigen-binding fragments of the present disclosure, in addition to exemplary antigen targets, are set forth in Tables A1-A3.
  • the antibody or antigen-binding fragment of an ADC disclosed herein may comprise amino acid sequences that are conservatively modified and/or homologous to the sequences listed in the tables above, so long as the ADC retains the ability to bind to its target cancer antigen (e.g., with a 42 ME150999036v.1 132043-01120 KD of less than 1x10 -8 M) and retains one or more functional properties of the ADCs disclosed herein (e.g., ability to internalize, bind to an antigen target, e.g., an antigen expressed on a tumor or other cancer cell, etc.).
  • target cancer antigen e.g., with a 42 ME150999036v.1 132043-01120 KD of less than 1x10 -8 M
  • one or more functional properties of the ADCs disclosed herein e.g., ability to internalize, bind to an antigen target, e.g., an antigen expressed on a tumor or other cancer cell, etc.
  • the antibody or antigen-binding fragment of an ADC disclosed herein further comprises human heavy and light chain constant domains or fragments thereof.
  • the antibody or antigen-binding fragment of the described ADCs may comprise a human IgG heavy chain constant domain (such as an IgG1) and a human kappa or lambda light chain constant domain.
  • the antibody or antigen-binding fragment of the described ADCs comprises a human immunoglobulin G subtype 1 (IgG1) heavy chain constant domain with a human Ig kappa light chain constant domain.
  • the target antigen for an ADC of the present disclosure is CD74.
  • the anti-CD74 antibody or antigen-binding fragment thereof is a VHmil x VK1aNQ antibody or antigen-binding fragment thereof.
  • the anti-CD74 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:1, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:2, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:10, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:11, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:12.
  • the anti-CD74 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:4, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:5, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:13, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:14, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:15.
  • HCDR1 consisting of SEQ ID NO:4
  • HCDR2 HCDR2
  • HCDR3 heavy chain CDR3
  • LCDR1 light chain CDR1
  • LCDR2 light chain CDR2
  • LCDR3 light chain CDR3
  • the anti-CD74 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:6, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:7, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:8; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:16, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:11, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:15.
  • HCDR1 consisting of SEQ ID NO:6
  • HCDR2 consisting of SEQ ID NO:7
  • HCDR3 HCDR3
  • LCDR1 light chain CDR1
  • LCDR2 light chain CDR2
  • LCDR3 light chain CDR3
  • the anti-CD74 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:9, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:5, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:13, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:14, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:15.
  • HCDR1 consisting of SEQ ID NO:9
  • HCDR2 consisting of SEQ ID NO:5
  • HCDR3 heavy chain CDR3
  • LCDR1 light chain CDR1
  • LCDR2 light chain CDR2
  • LCDR3 light chain CDR3
  • the anti-CD74 antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:17, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:18. In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises the heavy chain variable region amino acid sequence of SEQ ID NO:17 and the light chain variable region amino acid sequence of SEQ ID NO:18, or sequences that are at least 95% identical to the disclosed sequences.
  • the anti-CD74 antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:17 and/or a light chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:18.
  • the anti-CD74 antibody or antigen-binding fragment thereof comprises the heavy chain amino acid sequence of SEQ ID NO:19 or a sequence that is at least 95% identical to SEQ ID NO:19, and the light chain amino acid sequence of SEQ ID NO:20 or a sequence that is at least 95% identical to SEQ ID NO:20.
  • the anti-CD74 antibody or antigen- binding fragment thereof comprises the heavy chain amino acid sequence of SEQ ID NO:19 and the light chain amino acid sequence of SEQ ID NO:20, or sequences that are at least 95% identical to the disclosed sequences.
  • the anti-CD74 antibody or antigen-binding fragment thereof has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:19 and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:20.
  • Residues in two or more polypeptides are said to "correspond" if the residues occupy an analogous position in the polypeptide structures. Analogous positions in two or more polypeptides can be determined by aligning the polypeptide sequences based on amino acid sequence or structural similarities. Those skilled in the art understand that it may be necessary to introduce gaps in either sequence to produce a satisfactory alignment.
  • amino acid substitutions are of single residues. Insertions usually will be on the order of from about 1 to about 20 amino acid residues, although considerably larger insertions may be tolerated as long as biological function is retained (e.g., binding to a target antigen).
  • Deletions usually range from about 1 to about 20 amino acid residues, although in some cases deletions may be much larger. Substitutions, deletions, insertions, or any combination thereof may be used to arrive at a final derivative or variant. Generally, these changes are done on a few amino acids to minimize the alteration of the molecule, particularly the immunogenicity and specificity of the antigen binding protein. However, larger changes may be tolerated in certain circumstances. Conservative substitutions can be made in accordance with the following chart depicted as Table B.
  • the variant may be designed such that the biological activity of the antigen binding protein is altered. For example, glycosylation sites may be altered or removed.
  • CD74-targeting antibodies such as VHmil x VK1aNQ, provided particularly improved drug:antibody ratio, aggregation level, stability (i.e., in vitro and in vivo stability), tumor targeting (i.e., cytotoxicity, potency), minimized off-target killing, and/or treatment efficacy.
  • linker in an ADC is stable extracellularly in a sufficient manner to be therapeutically effective.
  • the linker is stable outside a cell, such that the ADC remains intact when present in extracellular conditions (e.g., prior to transport or delivery into a cell).
  • the term “intact,” used in the context of an ADC, means that the antibody or antigen-binding fragment remains attached to the drug moiety (e.g., the antineoplastic payloads, such as BH3 mimetics).
  • “stable,” in the context of a linker or ADC comprising a linker, means that no more than 20%, no more than about 15%, no more than about 10%, no more than about 5%, no more than about 3%, or no more than about 1% of the linkers (or any percentage in between) in a sample of ADC are cleaved (or in the case of an overall ADC are otherwise not intact) when the ADC is present in extracellular conditions.
  • the linkers and/or ADCs disclosed herein are stable compared to alternate linkers and/or ADCs with alternate linkers and/or antineoplastic payloads, such as BH3 mimetics, topoisomerase 1 inhibitors, or anti-mitotic drugs.
  • the ADCs disclosed herein can remain intact for more than about 48 hours, more than 60 hours, more than about 72 hours, more than about 84 hours, or more than about 96 hours.
  • Whether a linker is stable extracellularly can be determined, for example, by including an ADC in plasma for a predetermined time period (e.g., 2, 4, 6, 8, 16, 24, 48, or 72 hours) and then quantifying the amount of free drug moiety present in the plasma. Stability may allow the ADC time to localize to target cancer cells and prevent the premature release of the drug moiety, which could lower the therapeutic index of the ADC by indiscriminately damaging both normal and cancer tissues.
  • the linker is stable outside of a target cell and releases the drug moiety from the ADC once inside of the cell, such that the drug can bind to its target.
  • an effective linker will: (i) maintain the specific binding properties of the antibody or antigen-binding fragment; (ii) allow delivery, e.g., intracellular delivery, of the drug moiety via stable attachment to the antibody or antigen-binding fragment; (iii) remain stable and intact until the ADC has been transported or delivered to its target site; and (iv) allow for the therapeutic effect, e.g., cytotoxic effect, of the drug moiety after cleavage or alternate release mechanism.
  • Linkers may impact the physico-chemical properties of an ADC.
  • a linker may be "cleavable” or “non-cleavable” (Ducry and Stump (2010) Bioconjugate Chem. 21:5-13).
  • Cleavable linkers are designed to release the drug moiety (e.g., antineoplastic payloads, such as BH3 mimetics, topoisomerase 1 inhibitors, or anti-mitotic drugs) when subjected to 46 ME150999036v.1 132043-01120 certain environment factors, e.g., when internalized into the target cell, whereas non-cleavable linkers generally rely on the degradation of the antibody or antigen-binding fragment itself.
  • a wavy line ( ) indicates the point of attachment of the partial structure to the rest of the molecule.
  • Dual linkers of the present disclosure are described in International PCT Application PCT/US2023/022990, which is incorporated herein by reference in its entirety.
  • an intermediate which is the precursor of the linker moiety, is reacted with the drug moiety (e.g., BH3 mimetics, such as a Mcl-1 inhibitor, a Bcl-2 inhibitor and/or a Bcl-xL inhibitor) under appropriate conditions.
  • the drug moiety e.g., BH3 mimetics, such as a Mcl-1 inhibitor, a Bcl-2 inhibitor and/or a Bcl-xL inhibitor
  • reactive groups are used on the drug and/or the intermediate or linker.
  • the product of the reaction between the drug and the intermediate, or the derivatized drug (drug plus linker) is subsequently reacted with the antibody or antigen-binding fragment under conditions that facilitate conjugation of the drug and intermediate or derivatized drug and antibody or antigen-binding fragment.
  • the intermediate or linker may first be reacted with the antibody or antigen-binding fragment, or a derivatized antibody or antigen-binding fragment, and then reacted with the drug or derivatized drug.
  • a number of different reactions are available for covalent attachment of the drug moiety and/or linker moiety to the antibody or antigen-binding fragment. This is often accomplished by reaction of one or more amino acid residues of the antibody or antigen-binding fragment, including the amine groups of lysine, the free carboxylic acid groups of glutamic acid and aspartic acid, the sulfhydryl groups of cysteine, and the various moieties of the aromatic amino acids.
  • non-specific covalent attachment may be undertaken using a carbodiimide reaction to link a carboxy (or amino) group on a drug moiety to an amino (or carboxy) group on an antibody or antigen-binding fragment.
  • bifunctional agents such as dialdehydes or imidoesters may also be used to link the amino group on a drug moiety to an amino group on an antibody or antigen-binding fragment.
  • drugs e.g., a BH3 mimetic
  • This method involves the periodate oxidation of a drug that contains glycol or hydroxy groups, thus forming an aldehyde which is then reacted with the binding agent.
  • Drug loading is represented by p (or 2a in ADCs of formula (I) of the present disclosure), and is also referred to herein as the drug-to-antibody ratio (DAR). Drug loading may range from 2 to 32 drug moieties per antibody or antigen-binding fragment.
  • a is an integer from 1 to 16. In some embodiments, a is an integer from 1 to 16, 1 to 15, 1 to 14, 1 to 13, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2.
  • a is an integer from 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, or 2 to 3. In some embodiments, a is an integer from 1 to 16. In some embodiments, a is an integer from 1 to 8. In some embodiments, a is an integer from 1 to 5. In some embodiments, a is an integer from 2 to 4. In some embodiments, a is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, a is 2. In some embodiments, a is 4. [161] Drug loading may be limited by the number of attachment sites on the antibody or antigen- binding fragment.
  • the linker moiety (L) of the ADC attaches to the antibody or antigen-binding fragment through a chemically active group on one or more amino acid residues on the antibody or antigen-binding fragment.
  • the linker may be attached to the antibody or antigen-binding fragment via a free amino, imino, hydroxyl, thiol, or carboxyl group (e.g., to the N- or C-terminus, to the epsilon amino group of one or more lysine residues, to the free carboxylic acid group of one or more glutamic acid or aspartic acid residues, or to the sulfhydryl group of one or more cysteine residues).
  • the site to which the linker is attached can be a natural residue in the amino acid sequence of the antibody or antigen-binding fragment, or it can be introduced into the antibody or antigen-binding fragment, e.g., by DNA recombinant technology (e.g., by introducing a cysteine residue into the amino acid sequence) or by protein biochemistry (e.g., by reduction, pH adjustment, or hydrolysis).
  • the number of drug moieties that can be conjugated to an antibody or antigen-binding fragment is limited by the number of free cysteine residues.
  • an antibody may have only one or a few cysteine thiol groups, or may have only one or a few sufficiently reactive thiol groups through which a linker may be attached.
  • antibodies do not contain many free and reactive cysteine thiol groups that may be linked to a drug moiety. Indeed, most cysteine thiol residues in antibodies are involved in either interchain 50 ME150999036v.1 132043-01120 or intrachain disulfide bonds. Conjugation to cysteines can therefore, in some embodiments, require at least partial reduction of the antibody.
  • an optimal drug:antibody ratio should increase potency of the ADC (by increasing the number of attached drug moieties per antibody) without destabilizing the antibody or antigen-binding fragment.
  • an optimal ratio may be 2, 4, 6, or 8.
  • an optimal ratio may be 2 or 4.
  • an antibody or antigen-binding fragment is exposed to reducing conditions prior to conjugation in order to generate one or more free cysteine residues.
  • An antibody in some embodiments, may be reduced with a reducing agent such as dithiothreitol (DTT) or tris(2- carboxyethyl)phosphine (TCEP), under partial or total reducing conditions, to generate reactive cysteine thiol groups.
  • DTT dithiothreitol
  • TCEP tris(2- carboxyethyl)phosphine
  • Unpaired cysteines may be generated through partial reduction with limited molar equivalents of TCEP, which can reduce the interchain disulfide bonds which link the light chain and heavy chain (one pair per H-L pairing) and the two heavy chains in the hinge region (two pairs per H-H pairing in the case of human IgG1) while leaving the intrachain disulfide bonds intact (Stefano et al. (2013) Methods Mol Biol. 1045:145-71).
  • disulfide bonds within the antibodies are reduced electrochemically, e.g., by employing a working electrode that applies an alternating reducing and oxidizing voltage.
  • This approach can allow for on-line coupling of disulfide bond reduction to an analytical device (e.g., an electrochemical detection device, an NMR spectrometer, or a mass spectrometer) or a chemical separation device (e.g., a liquid chromatograph (e.g., an HPLC) or an electrophoresis device (see, e.g., US 2014/0069822)).
  • an antibody is subjected to denaturing conditions to reveal reactive nucleophilic groups on amino acid residues, such as cysteine.
  • the drug loading of an ADC may be controlled in different ways, e.g., by: (i) limiting the molar excess of drug-linker intermediate or linker reagent relative to antibody; (ii) limiting the conjugation reaction time or temperature; (iii) partial or limiting reductive conditions for cysteine thiol modification; and/or (iv) engineering by recombinant techniques the amino acid sequence of the antibody such that the number and position of cysteine residues is modified for control of the number and/or position of linker-drug attachments.
  • free cysteine residues are introduced into the amino acid sequence of the antibody or antigen-binding fragment.
  • cysteine engineered antibodies can be prepared wherein one or more amino acids of a parent antibody are replaced with a cysteine amino acid. Any form of antibody may be so engineered, i.e. mutated.
  • a parent Fab antibody fragment may be engineered to form a cysteine engineered Fab referred to as a "ThioFab.”
  • a parent monoclonal antibody may be engineered to form a "ThioMab.”
  • a single site mutation yields a single engineered cysteine residue in a ThioFab, whereas a single site mutation yields two engineered cysteine residues in a ThioMab, due to the dimeric nature of the IgG antibody.
  • DNA 51 ME150999036v.1 132043-01120 encoding an amino acid sequence variant of the parent polypeptide can be prepared by a variety of methods known in the art (see, e.g., the methods described in WO 2006/034488). These methods include, but are not limited to, preparation by site-directed (or oligonucleotide-mediated) mutagenesis, PCR mutagenesis, and cassette mutagenesis of an earlier prepared DNA encoding the polypeptide. Variants of recombinant antibodies may also be constructed by restriction fragment manipulation or by overlap extension PCR with synthetic oligonucleotides.
  • ADCs of Formula (1) include, but are not limited to, antibodies that have 1, 2, 3, or 4 engineered cysteine amino acids (Lyon et al. (2012) Methods Enzymol. 502:123-38).
  • one or more free cysteine residues are already present in an antibody or antigen-binding fragment, without the use of engineering, in which case the existing free cysteine residues may be used to conjugate the antibody or antigen-binding fragment to a drug moiety.
  • the resulting product can be a mixture of ADC compounds with a distribution of one or more drug moieties attached to each copy of the antibody or antigen-binding fragment in the mixture.
  • the drug loading in a mixture of ADCs resulting from a conjugation reaction ranges from 1 to 16 drug moieties attached per antibody or antigen-binding fragment.
  • the average number of drug moieties per antibody or antigen- binding fragment may be calculated by any conventional method known in the art, e.g., by mass spectrometry (e.g., liquid chromatography-mass spectrometry (LC-MS)) and/or high-performance liquid chromatography (e.g., HIC-HPLC).
  • mass spectrometry e.g., liquid chromatography-mass spectrometry (LC-MS)
  • HIC-HPLC high-performance liquid chromatography
  • the average number of drug moieties per antibody or antigen-binding fragment is determined by liquid chromatography-mass spectrometry (LC-MS).
  • the average number of drug moieties per antibody or antigen-binding fragment is from about 1.5 to about 3.5, about 2.5 to about 4.5, about 3.5 to about 5.5, about 4.5 to about 6.5, about 5.5 to about 7.5, about 6.5 to about 8.5, or about 7.5 to about 9.5. In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is from about 2 to about 4, about 3 to about 5, about 4 to about 6, about 5 to about 7, about 6 to about 8, about 7 to about 9, about 2 to about 8, or about 4 to about 8. [167] In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is about 2.
  • the average number of drug moieties per antibody or antigen-binding fragment is about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2, about 2.1, about 2.2, about 2.3, about 2.4, or about 2.5. In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is 2. [168] In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is about 4.
  • the average number of drug moieties per antibody or antigen-binding fragment is about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, about 4, about 4.1, 52 ME150999036v.1 132043-01120 about 4.2, about 4.3, about 4.4, or about 4.5. In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is 4. [169] In some embodiments, the term “about,” as used with respect to the average number of drug moieties per antibody or antigen-binding fragment, means plus or minus 20%, 15%, 10%, 5%, or 1%. In one embodiment, the term “about” refers to a range of values which are 10% more or less than the specified value.
  • the term “about” refers to a range of values which are 5% more or less than the specified value. In another embodiment, the term “about” refers to a range of values which are 1% more or less than the specified value.
  • Individual ADC compounds, or “species,” may be identified in the mixture by mass spectroscopy and separated by, e.g., UPLC or HPLC, e.g. hydrophobic interaction chromatography (HIC-HPLC). In some embodiments, a homogeneous or nearly homogenous ADC product with a single loading value may be isolated from the conjugation mixture, e.g., by electrophoresis or chromatography.
  • higher drug loading may cause aggregation, insolubility, toxicity, or loss of cellular permeability of certain antibody-drug conjugates. Higher drug loading may also negatively affect the pharmacokinetics (e.g., clearance) of certain ADCs.
  • lower drug loading e.g., p ⁇ 2
  • the drug loading for an ADC of the present disclosure ranges from about 2 to about 16, about 2 to about 10, about 2 to about 8; from about 2 to about 6; from about 2 to about 5; from about 3 to about 5; from about 2 to about 4; or from about 4 to about 8.
  • a drug loading and/or an average drug loading of about 2 is achieved, e.g., using partial reduction of intrachain disulfides on the antibody or antigen-binding fragment, and provides beneficial properties.
  • a drug loading and/or an average drug loading of about 4 or about 6 or about 8 is achieved, e.g., using partial reduction of intrachain disulfides on the antibody or antigen-binding fragment, and provides beneficial properties.
  • a drug loading and/or an average drug loading of less than about 2 may result in an unacceptably high level of unconjugated antibody species, which can compete with the ADC for binding to a target antigen and/or provide for reduced treatment efficacy.
  • a drug loading and/or average drug loading of more than about 16 may result in an unacceptably high level of product heterogeneity and/or ADC aggregation.
  • a drug loading and/or an average drug loading of more than about 16 may also affect stability of the ADC, due to loss of one or more chemical bonds required to stabilize the antibody or antigen-binding fragment.
  • the present disclosure includes methods of producing the described ADCs. Briefly, the ADCs comprise an antibody or antigen-binding fragment as the antibody or antigen-binding fragment, a drug moiety (e.g., a BH3 mimetic), and a linker that joins the drug moiety and the antibody or antigen-binding fragment.
  • the ADCs can be prepared using a linker having reactive functionalities for covalently attaching to the drug moiety and to the antibody or antigen- 53 ME150999036v.1 132043-01120 binding fragment.
  • the antibody or antigen-binding fragment is functionalized to prepare a functional group that is reactive with a linker or a drug-linker intermediate.
  • a cysteine thiol of an antibody or antigen-binding fragment can form a bond with a reactive functional group of a linker or a drug-linker intermediate to make an ADC.
  • an antibody or antigen-binding fragment is prepared with bacterial transglutaminase (BTG) - reactive glutamines specifically functionalized with an amine containing cyclooctyne BCN (N-[(1R,8S,9s)-Bicyclo[6.1.0]non-4-yn-9-ylmethyloxycarbonyl]-1,8-diamino-3,6-dioxaoctane) moiety.
  • BCG transglutaminase
  • BCN cyclooctyne
  • site-specific conjugation of a linker or a drug-linker intermediate to a BCN moiety of an antibody or antigen-binding fragment is performed, e.g., as described and exemplified herein.
  • an ADC is produced by contacting an antibody or antigen-binding fragment with a linker and a drug moiety (e.g., a BH3 mimetic) in a sequential manner, such that the antibody or antigen-binding fragment is covalently linked to the linker first, and then the pre-formed antibody-linker intermediate reacts with the drug moiety.
  • a linker and a drug moiety e.g., a BH3 mimetic
  • an ADC is produced by contacting an antibody or antigen-binding fragment with a linker-drug compound pre-formed by reacting a linker with a drug moiety.
  • the pre-formed linker-drug compound may or may not be subjected to a purification step prior to contacting the antibody or antigen-binding fragment.
  • the antibody or antigen-binding fragment contacts the linker and the drug moiety in one reaction mixture, allowing simultaneous formation of the covalent bonds between the antibody or antigen-binding fragment and the linker, and between the linker and the drug moiety.
  • This method of producing ADCs may include a reaction, wherein the antibody or antigen-binding fragment contacts the antibody or antigen-binding fragment prior to the addition of the linker to the reaction mixture, and vice versa.
  • an ADC is produced by reacting an antibody or antigen-binding fragment with a linker joined to a drug moiety, such as a BH3 mimetic, under conditions that allow conjugation.
  • the ADCs prepared according to the methods described above may be subjected to a purification step.
  • the purification step may involve any biochemical methods known in the art for purifying proteins, or any combination of methods thereof.
  • compositions described herein e.g., the disclosed ADC compounds and compositions, in treating a subject for a disorder, e.g., a cancer.
  • compositions e.g., ADCs
  • ADCs may be administered alone or in combination with at least one additional inactive and/or active agent, e.g., at least one additional therapeutic agent, and may be administered in any pharmaceutically acceptable formulation, dosage, and dosing regimen.
  • Treatment efficacy may be evaluated for toxicity as well as indicators of efficacy and adjusted accordingly.
  • Efficacy measures include, but are not limited to, a cytostatic and/or cytotoxic effect observed in vitro or in vivo, reduced tumor volume, tumor growth inhibition, and/or prolonged survival. [177] Methods of determining whether an ADC exerts a cytostatic and/or cytotoxic effect on a cell are known.
  • the cytotoxic or cytostatic activity of an ADC can be measured by, e.g., exposing mammalian cells expressing a target antigen of the ADC in a cell culture medium; culturing the cells for a period from about 6 hours to about 6 days; and measuring cell viability (e.g., using a CellTiter-Glo® (CTG) or MTT cell viability assay).
  • CCG CellTiter-Glo®
  • MTT cell viability assay Cell-based in vitro assays may also be used to measure viability (proliferation), cytotoxicity, and induction of apoptosis (caspase activation) of the ADC.
  • Necrosis is typically accompanied by increased permeability of the plasma membrane, swelling of the cell, and rupture of the plasma membrane.
  • Apoptosis can be quantitated, for example, by measuring DNA fragmentation.
  • Commercial photometric methods for the quantitative in vitro determination of DNA fragmentation are available. Examples of such assays, including TUNEL (which detects incorporation of labeled nucleotides in fragmented DNA) and ELISA-based assays, are described in Biochemica (1999) 2:34-7 (Roche Molecular Biochemicals).
  • Apoptosis may also be determined by measuring morphological changes in a cell.
  • loss of plasma membrane integrity can be determined by measuring uptake of certain dyes (e.g., a fluorescent dye such as, for example, acridine orange or ethidium bromide).
  • a fluorescent dye such as, for example, acridine orange or ethidium bromide.
  • a method for measuring apoptotic cell number has been described by Duke and Cohen, Current Protocols in Immunology (Coligan et al., eds. (1992) pp. 3.17.1-3.17.16).
  • Cells also can be labeled with a DNA dye (e.g., acridine orange, ethidium bromide, or propidium iodide) and the cells observed for chromatin condensation and margination along the inner nuclear membrane.
  • Apoptosis may also be determined, in some embodiments, by screening for caspase activity.
  • a Caspase-Glo® Assay can be used to measure activity of caspase-3 and caspase-7.
  • the assay provides a luminogenic caspase-3/7 substrate in a reagent optimized for caspase activity, luciferase activity, and cell lysis.
  • adding Caspase-Glo® 3/7 Reagent in an “add-mix-measure” format may result in cell lysis, followed by caspase cleavage of the substrate and generation of a “glow-type” luminescent signal, produced by luciferase.
  • luminescence may be proportional to the amount of caspase activity present, and can serve as an indicator of apoptosis.
  • Other morphological changes that can be measured to determine 55 ME150999036v.1 132043-01120 apoptosis include, e.g., cytoplasmic condensation, increased membrane blebbing, and cellular shrinkage. Determination of any of these effects on cancer cells indicates that an ADC is useful in the treatment of cancers.
  • Cell viability may be measured, e.g., by determining in a cell the uptake of a dye such as neutral red, trypan blue, Crystal Violet, or ALAMARTM blue (see, e.g., Page et al.
  • Cell viability may also be measured, e.g., by quantifying ATP, an indicator of metabolically active cells.
  • in vitro potency and/or cell viability of prepared ADCs or antineoplastic payloads such as BH3 mimetic compounds (e.g., MCl-1 inhibitor, Bcl-xL inhibitor or Bcl-2 inhibitor) or topoisomerase 1 inhibitors (e.g., topotecan, exatecan, deruxtecan or SN-38) or anti- mitotic drugs (e.g., monomethyl auristatin E (MMAE) or a taxane) may be assessed using a CellTiter- Glo® (CTG) cell viability assay, as described in the examples provided herein.
  • CTG CellTiter- Glo®
  • the single reagent (CellTiter-Glo® Reagent) is added directly to cells cultured in serum-supplemented medium.
  • the addition of reagent results in cell lysis and generation of a luminescent signal proportional to the amount of ATP present.
  • the amount of ATP is directly proportional to the number of cells present in culture.
  • Cell viability may also be measured, e.g., by measuring the reduction of tetrazolium salts.
  • in vitro potency and/or cell viability of prepared ADCs or antineoplastic payloads may be assessed using an MTT cell viability assay, as described in the examples provided herein.
  • MTT cell viability assay in some embodiments, the yellow tetrazolium MTT (3-(4, 5-dimethylthiazolyl-2)-2,5- diphenyltetrazolium bromide) is reduced by metabolically active cells, in part by the action of dehydrogenase enzymes, to generate reducing equivalents such as NADH and NADPH.
  • the present disclosure features a method of killing, inhibiting or modulating the growth of a cancer cell or tissue by disrupting the expression and/or activity of Bcl-2 family protein (e.g., Mcl-1, Bcl-2 and/or Bcl-xL) and/or one or more upstream modulators or downstream targets thereof.
  • Bcl-2 family protein e.g., Mcl-1, Bcl-2 and/or Bcl-xL
  • the method may be used with any subject where disruption of Bcl-2 family protein expression and/or activity provides a therapeutic benefit.
  • Subjects that may benefit from disrupting Bcl-2 family protein expression and/or activity include, but are not limited to, those having or at risk of having a cancer such as a tumor or a hematological cancer.
  • the cancer is a breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular cancer, lymphoblastic leukemia including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B- 56 ME150999036v.1 132043-01120 cell origin, melanoma, myelogenous leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, stomach cancer, colon cancer, or head and neck cancer.
  • the cancer is acute myeloid leukemia, multiple myeloma, or B-cell lymphoma.
  • the disclosed ADCs may be administered in any cell or tissue that expresses CD74, such as a CD74-expressing cancer cell or tissue.
  • An exemplary embodiment includes a method of killing a CD74-expressing cancer cell or tissue. The method may be used with any cell or tissue that expresses CD74, such as a cancerous cell or a metastatic lesion.
  • Non-limiting examples of CD74-expressing cancers include a breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular cancer, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myelogenous leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or spleen cancer.
  • the CD74-expressing cancer is a hematological cancer.
  • the CD74-expressing cancer is chronic lymphocytic leukemia (CLL), follicular lymphoma, mantle cell lymphoma, diffuse large B-cell lymphoma, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myelomonocytic leukemia (CMML), acute monocytic leukemia (AMoL), Hodgkin's lymphoma, non-Hodgkin's lymphoma or myelodysplasia syndrome (MDS).
  • CLL chronic lymphocytic leukemia
  • ALL acute lymphoblastic leukemia
  • AML acute myeloid leukemia
  • CLL chronic lymphocytic leukemia
  • CML chronic myelogenous leukemia
  • CML chronic myelomonocy
  • the CD74-expressing cancer is acute myeloid leukemia, multiple myeloma, or B-cell lymphoma.
  • Exemplary methods include the steps of contacting a cell with an ADC, as described herein, in an effective amount, i.e., an amount sufficient to kill the cell.
  • the method can be used on cells in culture, e.g., in vitro, in vivo, ex vivo, or in situ.
  • cells that express CD74 e.g., cells collected by biopsy of a tumor or metastatic lesion; cells from an established cancer cell line; or recombinant cells
  • the method will result in killing of cells expressing CD74, including in particular cancer cells expressing CD74.
  • the ADC can be administered to a subject by any suitable administration route (e.g., intravenous, subcutaneous, or direct contact with a tumor tissue) to have an effect in vivo.
  • This approach can be used for antibodies targeting cell surface antigens (e.g., CD74).
  • the in vivo effect of a disclosed ADC therapeutic composition can be evaluated in a suitable animal model.
  • xenogeneic cancer models can be used, wherein cancer explants or passaged xenograft tissues are introduced into immune compromised animals, such as nude or SCID mice (Klein et al. (1997) Nature Med. 3:402-8).
  • Efficacy may be predicted using assays that measure inhibition of tumor formation, tumor regression or metastasis, and the like. 57 ME150999036v.1 132043-01120 [187] In vivo assays that evaluate the promotion of tumor death by mechanisms such as apoptosis may also be used. In some embodiments, xenografts from tumor bearing mice treated with the therapeutic composition can be examined for the presence of apoptotic foci and compared to untreated control xenograft-bearing mice. The extent to which apoptotic foci are found in the tumors of the treated mice provides an indication of the therapeutic efficacy of the composition. [188] Further provided herein are methods of treating a disorder, e.g., a cancer.
  • compositions described herein can be administered to a non-human mammal or human subject for therapeutic purposes.
  • the therapeutic methods include administering to a subject having or suspected of having a cancer a therapeutically effective amount of a composition comprising an ADC with a targeting antibody that binds to an antigen (e.g., CD74) (1) expressed on a cancer cell, (2) is accessible to binding, and/or (3) is localized or predominantly expressed on a cancer cell surface as compared to a non-cancer cell.
  • an antigen e.g., CD74
  • An exemplary embodiment is a method of treating a subject having or suspected of having a cancer, comprising administering to the subject a therapeutically effective amount of a composition disclosed herein, e.g., an ADC, composition, or pharmaceutical composition (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein).
  • a composition disclosed herein e.g., an ADC, composition, or pharmaceutical composition
  • the cancer expresses a target antigen.
  • the target antigen is CD74.
  • the cancer is a tumor or a hematological cancer.
  • the cancer is a breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular cancer, lymphoblastic leukemia including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myelogenous leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, stomach cancer, colon cancer, or head and neck cancer.
  • the cancer is acute myeloid leukemia, multiple myeloma, or B-cell lymphoma.
  • Another exemplary embodiment is a method of delivering antineoplastic payloads (e.g. BH3 mimetics) to a cell expressing CD74, comprising conjugating the antineoplastic payloads (e.g. BH3 mimetics) to an antibody that immunospecifically binds to a CD74 epitope and exposing the cell to the ADC.
  • antineoplastic payloads e.g. BH3 mimetics
  • Exemplary cancer cells that express CD74 for which the ADCs of the present disclosure are indicated include hematological cancer cells.
  • the present disclosure further provides methods of reducing or inhibiting growth of a tumor (e.g., a CD74-expressing tumor), comprising administering a therapeutically effective amount of an ADC or composition comprising an ADC.
  • a tumor e.g., a CD74-expressing tumor
  • the treatment is sufficient to reduce or inhibit the growth of the patient's tumor, reduce the number or size of metastatic lesions, reduce tumor load, reduce primary tumor load, reduce invasiveness, prolong survival time, and/or maintain or improve the quality of life.
  • the tumor is 58 ME150999036v.1 132043-01120 resistant or refractory to treatment with the antibody or antigen-binding fragment of the ADC (e.g., an anti-CD74 antibody or antigen-binding fragment) when administered alone, and/or the tumor is resistant or refractory to treatment with a BH3 mimetic drug moiety when administered alone.
  • An exemplary embodiment is a method of reducing or inhibiting the growth of a tumor in a subject, comprising administering to the subject a therapeutically effective amount of an ADC, composition, or pharmaceutical composition (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein).
  • the tumor expresses a target antigen.
  • the target antigen is CD74.
  • the tumor is a breast cancer, gastric cancer, bladder cancer, brain cancer, cervical cancer, colorectal cancer, esophageal cancer, hepatocellular cancer, melanoma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, or spleen cancer.
  • administration of the ADC, composition, or pharmaceutical composition reduces or inhibits the growth of the tumor by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%, as compared to growth in the absence of treatment.
  • Another exemplary embodiment is a method of delaying or slowing the growth of a tumor in a subject, comprising administering to the subject a therapeutically effective amount of an ADC, composition, or pharmaceutical composition (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein).
  • the tumor expresses a target antigen.
  • the target antigen is CD74.
  • the tumor is a breast cancer, gastric cancer, bladder cancer, brain cancer, cervical cancer, colorectal cancer, esophageal cancer, hepatocellular cancer, melanoma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, or spleen cancer.
  • administration of the ADC, composition, or pharmaceutical composition delays or slows the growth of the tumor by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%, as compared to growth in the absence of treatment.
  • An exemplary embodiment is a method of reducing or inhibiting the growth of a hematological cancer in a subject, comprising administering to the subject a therapeutically effective amount of an ADC, composition, or pharmaceutical composition (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein).
  • the hematological cancer expresses a target antigen.
  • the target antigen is CD74.
  • the hematological cancer is chronic lymphocytic leukemia (CLL), follicular lymphoma, mantle cell lymphoma, diffuse large B-cell lymphoma, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous 59 ME150999036v.1 132043-01120 leukemia (CML), chronic myelomonocytic leukemia (CMML), acute monocytic leukemia (AMoL), Hodgkin's lymphoma, non-Hodgkin's lymphoma or myelodysplasia syndrome (MDS).
  • CLL chronic lymphocytic leukemia
  • ALL acute lymphoblastic leukemia
  • AML acute myeloid leukemia
  • CLL chronic lymphocytic leukemia
  • administration of the ADC, composition, or pharmaceutical composition reduces or inhibits the growth of the hematological cancer by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%, as compared to growth in the absence of treatment.
  • the present disclosure further provides methods of reducing or slowing the expansion of a cancer cell population (e.g., a CD74-expressing tumor), comprising administering a therapeutically effective amount of an ADC or composition comprising an ADC.
  • An exemplary embodiment is a method of reducing or slowing the expansion of a cancer cell population in a subject, comprising administering to the subject a therapeutically effective amount of an ADC, composition, or pharmaceutical composition (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein).
  • the cancer cell population expresses a target antigen.
  • the target antigen is CD74.
  • the cancer cell population is from a tumor or a hematological cancer.
  • the cancer cell population is from a breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular cancer, lymphoblastic leukemia including acute lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myelogenous leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer.
  • the cancer cell population is from acute myeloid leukemia, multiple myeloma, or B-cell lymphoma.
  • administration of the ADC, composition, or pharmaceutical composition reduces the cancer cell population by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%, as compared to the population in the absence of treatment.
  • administration of the ADC, composition, or pharmaceutical composition slows the expansion of the cancer cell population by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%, as compared to expansion in the absence of treatment.
  • Also provided herein are methods of determining whether a subject having or suspected of having a cancer will be responsive to treatment with the disclosed ADCs and compositions.
  • An exemplary embodiment is a method of determining whether a subject having or suspected of having a cancer will be responsive to treatment with an ADC, composition, or pharmaceutical composition 60 ME150999036v.1 132043-01120 (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein) by providing a biological sample from the subject; contacting the sample with the ADC; and detecting binding of the ADC to cancer cells in the sample.
  • the sample is a tissue biopsy sample, a blood sample, or a bone marrow sample.
  • the method comprises providing a biological sample from the subject; contacting the sample with the ADC; and detecting one or more markers of cancer cell death in the sample (e.g., increased expression of one or more apoptotic markers, reduced expansion of a cancer cell population in culture, etc.).
  • markers of cancer cell death in the sample e.g., increased expression of one or more apoptotic markers, reduced expansion of a cancer cell population in culture, etc.
  • An exemplary embodiment is an ADC, composition, or pharmaceutical composition (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein) for use in treating a subject having or suspected of having a cancer (e.g., a CD74-expressing cancer).
  • Another exemplary embodiment is a use of an ADC, composition, or pharmaceutical composition (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein) in treating a subject having or suspected of having a cancer (e.g., a CD74-expressing cancer).
  • Another exemplary embodiment is a use of an ADC, composition, or pharmaceutical composition (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein) in a method of manufacturing a medicament for treating a subject having or suspected of having a cancer (e.g., a CD74-expressing cancer).
  • ADCs of the present disclosure may be administered to a non-human mammal expressing an antigen with which the ADC is capable of binding for veterinary purposes or as an animal model of human disease. Regarding the latter, such animal models may be useful for evaluating the therapeutic efficacy of the disclosed ADCs (e.g., testing of dosages and time courses of administration).
  • the therapeutic compositions used in the practice of the foregoing methods may be formulated into pharmaceutical compositions comprising a pharmaceutically acceptable carrier suitable for the desired delivery method.
  • An exemplary embodiment is a pharmaceutical composition
  • a pharmaceutical composition comprising an ADC of the present disclosure and a pharmaceutically acceptable carrier, e.g., one suitable for a chosen means of administration, e.g., intravenous administration.
  • the pharmaceutical composition may also comprise one or more additional inactive and/or therapeutic agents that are suitable for treating or preventing, for example, a cancer (e.g., a standard-of-care agent, etc.).
  • the pharmaceutical composition may also comprise one or more carrier, excipient, and/or stabilizer components, and the like. Methods of formulating such pharmaceutical compositions and suitable formulations are known in the art (see, e.g., "Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, PA).
  • Suitable carriers include any material that, when combined with the therapeutic composition, retains the anti-tumor function of the therapeutic composition and is generally non-reactive with the patient's immune system.
  • Pharmaceutically acceptable carriers include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. Examples of pharmaceutically acceptable carriers include one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol, mesylate salt, and the like, as well as combinations thereof.
  • isotonic agents are included, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition.
  • Pharmaceutically acceptable carriers may further comprise minor amounts of auxiliary substances such as wetting or emulsifying agents, preservatives or buffers, which enhance the shelf life or effectiveness of the ADC.
  • a pharmaceutical composition of the present disclosure can be administered by a variety of methods known in the art. The route and/or mode of administration may vary depending upon the desired results. In some embodiments, the therapeutic formulation is solubilized and administered via any route capable of delivering the therapeutic composition to the cancer site.
  • routes of administration include, but are not limited to, parenteral (e.g., intravenous, subcutaneous), intraperitoneal, intramuscular, intratumor, intradermal, intraorgan, orthotopic, and the like.
  • the administration is intravenous, subcutaneous, intraperitoneal, or intramuscular.
  • the pharmaceutically acceptable carrier should be suitable for the route of administration, e.g., intravenous or subcutaneous administration (e.g., by injection or infusion).
  • the active compound(s) i.e., the ADC and/or any additional therapeutic agent, may be coated in a material to protect the compound(s) from the action of acids and other natural conditions that may inactivate the compound(s).
  • the therapeutic compositions disclosed herein may be sterile and stable under the conditions of manufacture and storage, and may be in a variety of forms. These include, for example, liquid, semi-solid, and solid dosage forms, such as liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, tablets, pills, powders, liposomes, and suppositories. The form depends on the intended mode of administration and therapeutic application.
  • the disclosed ADCs can be incorporated into a pharmaceutical composition suitable for parenteral administration.
  • the injectable solution may be composed of either a liquid or lyophilized dosage form in a flint or amber vial, ampule, or pre-filled syringe, or other known delivery or storage device.
  • one or more of the ADCs or pharmaceutical compositions is supplied as a dry sterilized lyophilized powder or water free concentrate in a hermetically sealed container and can be reconstituted (e.g., with water or saline) to the appropriate concentration for administration to a subject.
  • a therapeutically effective amount or efficacious amount of a disclosed composition e.g., a disclosed ADC, is employed in the pharmaceutical compositions of the present 62 ME150999036v.1 132043-01120 disclosure.
  • compositions e.g., one comprising an ADC
  • the composition may be formulated into a pharmaceutically acceptable dosage form by conventional methods known in the art. Dosages and administration protocols for the treatment of cancers using the foregoing methods will vary with the method and the target cancer, and will generally depend on a number of other factors appreciated in the art. [207] Dosage regimens for compositions disclosed herein, e.g., those comprising ADCs alone or in combination with at least one additional inactive and/or active therapeutic agent, may be adjusted to provide the optimum desired response (e.g., a therapeutic response).
  • a single bolus of one or both agents may be administered at one time, several divided doses may be administered over a predetermined period of time, or the dose of one or both agents may be proportionally increased or decreased as indicated by the exigencies of the therapeutic situation.
  • treatment involves single bolus or repeated administration of the ADC preparation via an acceptable route of administration.
  • the ADC is administered to the patient daily, weekly, monthly, or any time period in between.
  • specific dosage regimens may be adjusted over time according to the individual’s need, and the professional judgment of the treating clinician.
  • Parenteral compositions may be formulated in dosage unit form for ease of administration and uniformity of dosage.
  • Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit contains a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.
  • Dosage values for compositions comprising an ADC and/or any additional therapeutic agent(s) may be selected based on the unique characteristics of the active compound(s), and the particular therapeutic effect to be achieved. A physician or veterinarian can start doses of the ADC employed in the pharmaceutical composition at levels lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.
  • effective doses of the compositions of the present disclosure, for the treatment of a cancer may vary depending upon many different factors, including means of administration, target site, physiological state of the patient, whether the patient is human or an animal, other medications administered, and whether treatment is prophylactic or therapeutic.
  • the selected dosage level may also depend upon a variety of pharmacokinetic factors including the activity of the particular compositions of the present disclosure employed, or the ester, salt, or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compositions employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors.
  • Treatment dosages may be titrated to optimize safety and efficacy.
  • Toxicity and therapeutic efficacy of compounds provided herein can be determined by standard pharmaceutical procedures in cell culture or in animal models. For example, LD50, ED50, EC50, and IC50 may be determined, and the dose ratio between toxic and therapeutic effects (LD50/ED50) may be calculated as the therapeutic index.
  • the data obtained from in vitro and in vivo assays can be used in estimating or formulating a range of dosage for use in humans.
  • the compositions and methods disclosed herein may initially be evaluated in xenogeneic cancer models (e.g., an NCI-H929 multiple myeloma mouse model).
  • an ADC or composition comprising an ADC is administered on a single occasion. In other embodiments, an ADC or composition comprising an ADC is administered on multiple occasions. Intervals between single dosages can be, e.g., daily, weekly, monthly, or yearly. Intervals can also be irregular, based on measuring blood levels of the administered agent (e.g., the ADC) in the patient in order to maintain a relatively consistent plasma concentration of the agent.
  • the dosage and frequency of administration of an ADC or composition comprising an ADC may also vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic applications, a relatively low dosage may be administered at relatively infrequent intervals over a long period of time. Some patients continue to receive treatment for the rest of their lives.
  • kits for use in the therapeutic and/or diagnostic applications described herein are also provided.
  • kits may comprise a carrier, package, or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) comprising one of the separate elements to be used in a method disclosed herein.
  • a label may be present on or with the container(s) to indicate that an ADC or composition within the kit is used for a specific therapy or non-therapeutic application, such as a prognostic, prophylactic, diagnostic, or laboratory application.
  • a label may also indicate directions for either in vivo or in vitro use, such as those described herein. Directions and or other information may also be included on an insert(s) or label(s), which is included with or on the kit.
  • the label may be on or associated with the container.
  • a label may be on a container when letters, numbers, or other characters forming the label are molded or etched into the container itself.
  • a label may be associated with a container when it is present within a receptacle or carrier that also holds the container, e.g., as a package insert.
  • the label may indicate 64 ME150999036v.1 132043-01120 that an ADC or composition within the kit is used for diagnosing or treating a condition, such as a cancer a described herein.
  • a kit comprises an ADC or composition comprising an ADC.
  • the kit further comprises one or more additional components, including but not limited to: instructions for use; other reagents, e.g., a therapeutic agent (e.g., a standard-of-care agent); devices, containers, or other materials for preparing the ADC for administration; pharmaceutically acceptable carriers; and devices, containers, or other materials for administering the ADC to a subject.
  • a therapeutic agent e.g., a standard-of-care agent
  • kits, or other materials for preparing the ADC for administration e.g., a standard-of-care agent
  • pharmaceutically acceptable carriers e.g., a standard-of-care agent
  • the kit comprises an ADC and instructions for use of the ADC in treating, preventing, and/or diagnosing a cancer.
  • ADCs Antibody-drug conjugates
  • BCL2 family members e.g., Bcl-xL
  • Antibody-drug conjugates that may not be sufficiently effective as monotherapy to treat cancer can be administered in combination with other therapeutic agents (including non-targeted and targeted therapeutic agents) or radiation therapy (including radioligand therapy) to provide therapeutic benefit.
  • other therapeutic agents including non-targeted and targeted therapeutic agents
  • radiation therapy including radioligand therapy
  • the ADCs described herein sensitize tumor cells to the treatment with other therapeutic agents (including standard of care chemotherapeutic agents to which the tumor cells may have developed resistance) and/or radiation therapy.
  • antibody drug conjugates described herein are administered to a subject having cancer in an amount effective to sensitize the tumor cells.
  • the term “sensitize” means that the treatment with ADC increases the potency or efficacy of the treatment with other therapeutic agents and/or radiation therapy against tumor cells. 3.
  • Combination Therapies [215] In some embodiments, the present disclosure provides methods of treatment wherein the antibody-drug conjugates disclosed herein are administered in combination with one or more (e.g., 1 or 2) additional therapeutic agents. Exemplary combination partners are disclosed herein. [216] In certain embodiments, a combination described herein comprises a PD-1 inhibitor.
  • the PD-1 inhibitor is chosen from PDR001 (Novartis), Nivolumab (Bristol-Myers Squibb), Pembrolizumab (Merck & Co), Pidilizumab (CureTech), MEDI0680 (Medimmune), REGN2810 (Regeneron), TSR-042 (Tesaro), PF-06801591 (Pfizer), BGB-A317 (Beigene), BGB-108 (Beigene), INCSHR1210 (Incyte), or AMP-224 (Amplimmune).
  • the PD-1 inhibitor is PDR001.
  • PDR001 is also known as Spartalizumab.
  • a combination described herein comprises a LAG-3 inhibitor.
  • the LAG-3 inhibitor is chosen from LAG525 (Novartis), BMS-986016 (Bristol- Myers Squibb), or TSR-033 (Tesaro). 65 ME150999036v.1 132043-01120 [218] In certain embodiments, a combination described herein comprises a TIM-3 inhibitor.
  • the TIM-3 inhibitor is MBG453 (Novartis), TSR-022 (Tesaro), LY-3321367 (Eli Lily), Sym23 (Symphogen), BGB-A425 (Beigene), INCAGN-2390 (Agenus), BMS-986258 (BMS), RO-7121661 (Roche), or LY-3415244 (Eli Lilly).
  • a combination described herein comprises a PDL1 inhibitor.
  • the PDL1 inhibitor is chosen from FAZ053 (Novartis), atezolizumab (Genentech), durvalumab (Astra Zeneca), or avelumab (Pfizer).
  • a combination described herein comprises a GITR agonist.
  • the GITR agonist is chosen from GWN323 (NVS), BMS-986156, MK-4166 or MK- 1248 (Merck), TRX518 (Leap Therapeutics), INCAGN1876 (Incyte/Agenus), AMG 228 (Amgen) or INBRX-110 (Inhibrx).
  • a combination described herein comprises an IAP inhibitor.
  • the IAP inhibitor comprises LCL161 or a compound disclosed in International Application Publication No. WO 2008/016893.
  • the combination comprises an mTOR inhibitor, e.g., RAD001 (also known as everolimus).
  • the combination comprises a HDAC inhibitor, e.g., LBH589. LBH589 is also known as panobinostat.
  • the combination comprises an IL-17 inhibitor, e.g., CJM112.
  • a combination described herein comprises an estrogen receptor (ER) antagonist.
  • the estrogen receptor antagonist is used in combination with a PD- 1 inhibitor, a CDK4/6 inhibitor, or both.
  • the combination is used to treat an ER positive (ER+) cancer or a breast cancer (e.g., an ER+ breast cancer).
  • the estrogen receptor antagonist is a selective estrogen receptor degrader (SERD).
  • SESDs are estrogen receptor antagonists which bind to the receptor and result in e.g., degradation or down-regulation of the receptor (Boer K. et al., (2017) Therapeutic Advances in Medical Oncology 9(7): 465-479).
  • ER is a hormone-activated transcription factor important for e.g., the growth, development and physiology of the human reproductive system. ER is activated by, e.g., the hormone estrogen (17beta estradiol).
  • the SERD is chosen from LSZ102, fulvestrant, brilanestrant, or elacestrant. [227] In some embodiments, the SERD comprises a compound disclosed in International Application Publication No. WO 2014/130310, which is hereby incorporated by reference in its entirety. [228] In some embodiments, the SERD comprises LSZ102.
  • LSZ102 has the chemical name: (E)-3- (4-((2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6-hydroxybenzo[b]thiophen-3-yl)oxy)phenyl)acrylic 66 ME150999036v.1 132043-01120 acid.
  • the SERD comprises fulvestrant (CAS Registry Number: 129453-61-8), or a compound disclosed in International Application Publication No. WO 2001/051056, which is hereby incorporated by reference in its entirety.
  • the SERD comprises elacestrant (CAS Registry Number: 722533-56-4), or a compound disclosed in U.S. Patent No.
  • Elacestrant is also known as RAD1901, ER-306323 or (6R)-6- ⁇ 2-[Ethyl( ⁇ 4-[2-(ethylamino)ethyl]phenyl ⁇ methyl)amino]-4-methoxyphenyl ⁇ - 5,6,7,8-tetrahydronaphthalen-2-ol.
  • Elacestrant is an orally bioavailable, non-steroidal combined selective estrogens receptor modulator (SERM) and a SERD.
  • SERM selective estrogens receptor modulator
  • Elacestrant is also disclosed, e.g., in Garner F et al., (2015) Anticancer Drugs 26(9):948-56.
  • the SERD is brilanestrant (CAS Registry Number: 1365888-06-7), or a compound disclosed in International Application Publication No. WO 2015/136017, which is incorporated by reference in its entirety. [229] In some embodiments, the SERD is chosen from RU 58668, GW7604, AZD9496, apeledoxifene, pipendoxifene, arzoxifene, OP-1074, or acolbifene, e.g., as disclosed in McDonell et al. (2015) Journal of Medicinal Chemistry 58(12) 4883-4887.
  • a combination described herein comprises an inhibitor of Cyclin- Dependent Kinases 4 or 6 (CDK4/6).
  • CDK4/6 Cyclin- Dependent Kinases 4 or 6
  • the CDK4/6 inhibitor is used in combination with a PD-1 inhibitor, an estrogen receptor (ER) antagonist, or both.
  • the combination is used to treat an ER positive (ER+) cancer or a breast cancer (e.g., an ER+ breast cancer).
  • the CDK4/6 inhibitor is chosen from ribociclib, abemaciclib (Eli Lilly), or palbociclib.
  • the CDK4/6 inhibitor comprises ribociclib (CAS Registry Number: 1211441-98-3), or a compound disclosed in U.S. Patent Nos. 8,415,355 and 8,685,980, which are incorporated by reference in their entirety.
  • the CDK4/6 inhibitor comprises a compound disclosed in International Application Publication No. WO 2010/020675 and U.S. Patent Nos. 8,415,355 and 8,685,980, which are incorporated by reference in their entirety.
  • the CDK4/6 inhibitor comprises ribociclib (CAS Registry Number: 1211441-98-3). Ribociclib is also known as LEE011, KISQALI®, or 7-cyclopentyl-N,N-dimethyl-2- ((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide.
  • the CDK4/6 inhibitor comprises abemaciclib (CAS Registry Number: 1231929-97-7).
  • Abemaciclib is also known as LY835219 or N-[5-[(4-Ethyl-1-piperazinyl)methyl]-2- pyridinyl]-5-fluoro-4-[4-fluoro-2-methyl-1-(1-methylethyl)-1H-benzimidazol-6-yl]-2- pyrimidinamine.
  • Abemaciclib is a CDK inhibitor selective for CDK4 and CDK6 and is disclosed, e.g., in Torres-Guzman R et al. (2017) Oncotarget 10.18632/oncotarget.17778.
  • the CDK4/6 inhibitor comprises palbociclib (CAS Registry Number: 571190-30-2).
  • Palbociclib is also known as PD-0332991, IBRANCE® or 6-Acetyl-8-cyclopentyl-5- methyl-2- ⁇ [5-(1-piperazinyl)-2-pyridinyl]amino ⁇ pyrido[2,3-d]pyrimidin-7(8H)-one.
  • Palbociclib inhibits CDK4 with an IC50 of 11nM, and inhibits CDK6 with an IC50 of 16nM, and is disclosed, e.g., in Finn et al.
  • a combination described herein comprises an inhibitor of chemokine (C-X-C motif) receptor 2 (CXCR2).
  • CXCR2 inhibitor is chosen from 6- chloro-3-((3,4-dioxo-2-(pentan-3-ylamino)cyclobut-1-en-1-yl)amino)-2-hydroxy-N-methoxy-N- methylbenzenesulfonamide, danirixin, reparixin, or navarixin.
  • the CSF-1/1R binding agent is chosen from an inhibitor of macrophage colony-stimulating factor (M-CSF), e.g., a monoclonal antibody or Fab to M-CSF (e.g., MCS110), a CSF-1R tyrosine kinase inhibitor (e.g., 4-((2-(((1R,2R)-2- hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide or BLZ945), a receptor tyrosine kinase inhibitor (RTK) (e.g., pexidartinib), or an antibody targeting CSF-1R (e.g., emactuzumab or FPA008).
  • M-CSF macrophage colony-stimulating factor
  • MCS110 monoclonal antibody or Fab to M-CSF
  • CSF-1R tyrosine kinase inhibitor e
  • the CSF-1/1R inhibitor is BLZ945.
  • the CSF-1/1R binding agent is MCS110.
  • the CSF-1/1R binding agent is pexidartinib.
  • a combination described herein comprises a c-MET inhibitor.
  • c- MET a receptor tyrosine kinase overexpressed or mutated in many tumor cell types, plays key roles in tumor cell proliferation, survival, invasion, metastasis, and tumor angiogenesis. Inhibition of c- MET may induce cell death in tumor cells overexpressing c-MET protein or expressing constitutively activated c-MET protein.
  • the c-MET inhibitor is chosen from capmatinib (INC280), JNJ-3887605, AMG 337, LY2801653, MSC2156119J, crizotinib, tivantinib, or golvatinib.
  • a combination described herein comprises a transforming growth factor beta (also known as TGF- ⁇ TGF ⁇ , TGFb, or TGF-beta, used interchangeably herein) inhibitor.
  • the TGF- ⁇ inhibitor is chosen from fresolimumab or XOMA 089.
  • a combination described herein comprises an adenosine A2a receptor (A2aR) antagonist (e.g., an inhibitor of A2aR pathway, e.g., an adenosine inhibitor, e.g., an inhibitor of A2aR or CD-73).
  • A2aR antagonist is used in combination with a PD-1 inhibitor, and one or more (e.g., two, three, four, five, or all) of a CXCR2 inhibitor, a CSF-1/1R binding agent, LAG-3 inhibitor, a GITR agonist, a c-MET inhibitor, or an IDO inhibitor.
  • the combination is used to treat a pancreatic cancer, a colorectal cancer, a gastric cancer, or a melanoma (e.g., a refractory melanoma).
  • the A2aR antagonist is chosen from PBF509 (NIR178) (Palobiofarma/Novartis), CPI444/V81444 (Corvus/Genentech), AZD4635/HTL-1071 (AstraZeneca/Heptares), Vipadenant (Redox/Juno), GBV-2034 (Globavir), AB928 (Arcus Biosciences), Theophylline, Istradefylline (Kyowa Hakko Kogyo), Tozadenant/SYN- 115 (Acorda), KW-6356 (Kyowa Hakko Kogyo), ST-4206 (Leadiant Biosciences), or 68 ME150999036v.1 132043
  • a combination described herein comprises an inhibitor of indoleamine 2,3-dioxygenase (IDO) and/or tryptophan 2,3-dioxygenase (TDO).
  • IDO indoleamine 2,3-dioxygenase
  • TDO tryptophan 2,3-dioxygenase
  • the IDO inhibitor is used in combination with a PD-1 inhibitor, and one or more (e.g., two, three, four, or all) of a TGF- ⁇ inhibitor, an A2aR antagonist, a CSF-1/1R binding agent, a c-MET inhibitor, or a GITR agonist.
  • the combination is used to treat a pancreatic cancer, a colorectal cancer, a gastric cancer, or a melanoma (e.g., a refractory melanoma).
  • the IDO inhibitor is chosen from (4E)-4-[(3-chloro-4-fluoroanilino)- nitrosomethylidene]-1,2,5-oxadiazol-3-amine (also known as epacadostat or INCB24360), indoximod (NLG8189), (1-methyl-D-tryptophan), ⁇ -cyclohexyl-5H-Imidazo[5,1-a]isoindole-5-ethanol (also known as NLG919), indoximod, BMS-986205 (formerly F001287).
  • a combination described herein comprises a Galectin, e.g., Galectin- 1 or Galectin-3, inhibitor.
  • the combination comprises a Galectin-1 inhibitor and a Galectin-3 inhibitor.
  • the combination comprises a bispecific inhibitor (e.g., a bispecific antibody molecule) targeting both Galectin-1 and Galectin-3.
  • the Galectin inhibitor is used in combination with one or more therapeutic agents described herein.
  • the Galectin inhibitor is chosen from an anti-Galectin antibody molecule, GR-MD-02 (Galectin Therapeutics), Galectin-3C (Mandal Med), Anginex, or OTX-008 (OncoEthix, Merck).
  • a combination described herein comprises an inhibitor of the MAP kinase pathway including ERK inhibitors, MEK inhibitors and RAF inhibitors.
  • a combination described herein comprises a MEK inhibitor.
  • the MEK inhibitor is chosen from Trametinib, selumetinib, AS703026, BIX 02189, BIX 02188, CI-1040, PD0325901, PD98059, U0126, XL-518, G-38963, or G02443714. [246] In some embodiments, the MEK inhibitor is trametinib.
  • Trametinib is also known as JTP- 74057, TMT212, N-(3- ⁇ 3-cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino]-6,8-dimethyl-2,4,7-trioxo- 3,4,6,7-tetrahydropyrido[4,3-d]pyrimidin-1(2H)-yl ⁇ phenyl)acetamide, or Mekinist (CAS Number 871700-17-3).
  • the MEK inhibitor comprises selumetinib which has the chemical name: (5-[(4-bromo-2-chlorophenyl)amino]-4-fluoro-N-(2-hydroxyethoxy)-1-methyl-1H- benzimidazole-6-carboxamide.
  • Selumetinib is also known as AZD6244 or ARRY 142886, e.g., as described in PCT Publication No. WO2003077914.
  • the MEK inhibitor comprises AS703026, BIX 02189 or BIX 02188.
  • the MEK inhibitor comprises 2-[(2-Chloro-4-iodophenyl)amino]-N- (cyclopropylmethoxy)-3,4-difluoro-benzamide (also known as CI-1040 or PD184352), e.g., as described in PCT Publication No. WO2000035436).
  • the MEK inhibitor comprises N-[(2R)-2,3-Dihydroxypropoxy]-3,4- difluoro-2-[(2-fluoro-4-iodophenyl)amino]- benzamide (also known as PD0325901), e.g., as described in PCT Publication No. WO2002006213).
  • the MEK inhibitor comprises 2’-amino-3’-methoxyflavone (also known as PD98059) which is available from Biaffin GmbH & Co., KG, Germany.
  • the MEK inhibitor comprises 2,3-bis[amino[(2- aminophenyl)thio]methylene]-butanedinitrile (also known as U0126), e.g., as described in US Patent No. 2,779,780).
  • the MEK inhibitor comprises XL-518 (also known as GDC-0973) which has a CAS No. 1029872-29-4 and is available from ACC Corp.
  • the MEK inhibitor comprises G-38963.
  • the MEK inhibitor comprises G02443714 (also known as AS703206) [256] Additional examples of MEK inhibitors are disclosed in WO 2013/019906, WO 03/077914, WO 2005/121142, WO 2007/04415, WO 2008/024725 and WO 2009/085983, the contents of which are incorporated herein by reference. Further examples of MEK inhibitors include, but are not limited to, 2,3-Bis[amino[(2-aminophenyl)thio]methylene]-butanedinitrile (also known as U0126 and described in US Patent No.
  • WO2003076424 vemurafenib (PLX-4032, CAS 918504- 65-1); (R)-3-(2,3-Dihydroxypropyl)-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3- d]pyrimidine-4,7(3H,8H)-dione (TAK-733, CAS 1035555-63-5); pimasertib (AS-703026, CAS 1204531-26-9); 2-(2-Fluoro-4-iodophenylamino)-N-(2-hydroxyethoxy)-1,5-dimethyl-6-oxo-1,6- dihydropyridine-3-carboxamide (AZD 8330); and 3,4-Difluoro-2-[(2-fluoro-4-iodophenyl)amino]-N- (2-hydroxyethoxy)-5-[(3-oxo-[1,2]o
  • a combination described herein comprises a RAF inhibitor.
  • RAF inhibitors include, but are not limited to, Vemurafenib (or Zelboraf®, PLX-4032, CAS 918504-65-1), GDC-0879, PLX-4720 (available from Symansis), Dabrafenib (or GSK2118436), LGX 818, CEP-32496, UI-152, RAF 265, Regorafenib (BAY 73-4506), CCT239065, or Sorafenib (or Sorafenib Tosylate, or Nexavar®).
  • the RAF inhibitor is Dabrafenib.
  • the RAF inhibitor is LXH254.
  • a combination described herein comprises an ERK inhibitor.
  • ERK inhibitors include, but are not limited to, LTT462, ulixertinib (BVD-523), LY3214996, GDC-0994, KO-947 and MK-8353.
  • the ERK inhibitor is LTT462.
  • LTT462 is 4-(3-amino-6-((1S,3S,4S)-3- fluoro-4-hydroxy ⁇ cyclohexyl)pyrazin-2-yl)-N-((S)-1-(3-bromo-5-fluorophenyl)-2- (methylamino) ⁇ ethyl)-2-fluorobenzamide and is the compound of the following structure: 70 ME150999036v.1 132043-01120 [264] The preparation of LTT462 is described in PCT patent application publication WO2015/066188. LTT462 is an inhibitor of extracellular signal-regulated kinases 1 and 2 (ERK 1/2).
  • a combination described herein comprises a taxane, a vinca alkaloid, a MEK inhibitor, an ERK inhibitor, or a RAF inhibitor.
  • a combination described herein comprises at least two inhibitors selected, independently, from a MEK inhibitor, an ERK inhibitor, and a RAF inhibitor.
  • a combination described herein comprises an anti-mitotic drug.
  • a combination described herein comprises a taxane.
  • Taxanes include, but are not limited to, docetaxel, paclitaxel, or cabazitaxel. In some embodiments, the taxane is docetaxel.
  • a combination described herein comprises a vinca alkaloid.
  • Vinca alkaloids include, but are not limited to, vincristine, vinblastine, and leurosine.
  • a combination described herein comprises a topoisomerase inhibitor.
  • Topoisomerase inhibitors include, but are not limited to, topotecan, irinotecan, camptothecin, diflomotecan, lamellarin D, ellipticines, etoposide (VP-16), teniposide, doxorubicin, daunorubicin, mitoxantrone, amsacrine, aurintricarboxylic acid, and HU-331.
  • a combination described herein includes an interleukin-1 beta (IL-1 ⁇ ) inhibitor.
  • the IL-1 ⁇ inhibitor is chosen from canakinumab, gevokizumab, Anakinra, or Rilonacept.
  • a combination described herein comprises an IL-15/IL-15Ra complex.
  • the IL-15/IL-15Ra complex is chosen from NIZ985 (Novartis), ATL-803 (Altor) or CYP0150 (Cytune).
  • a combination described herein comprises a mouse double minute 2 homolog (MDM2) inhibitor.
  • the human homolog of MDM2 is also known as HDM2.
  • an MDM2 inhibitor described herein is also known as a HDM2 inhibitor.
  • the MDM2 inhibitor is chosen from HDM201 or CGM097.
  • the MDM2 inhibitor comprises (S)-1-(4-chlorophenyl)-7-isopropoxy-6- methoxy-2-(4-(methyl(((1r,4S)-4-(4-methyl-3-oxopiperazin-1-yl)cyclohexyl)methyl)amino)phenyl)- 1,2-dihydroisoquinolin-3(4H)-one (also known as CGM097) or a compound disclosed in PCT Publication No. WO 2011/076786 to treat a disorder, e.g., a disorder described herein).
  • a therapeutic agent disclosed herein is used in combination with CGM097.
  • a combination described herein comprises a hypomethylating agent (HMA).
  • HMA hypomethylating agent
  • the HMA is chosen from decitabine or azacitidine.
  • a combination described herein comprises a glucocorticoid.
  • the glucocorticoid is dexamethasone.
  • a combination described herein comprises asparaginase.
  • a combination described herein comprises an inhibitor acting on any pro-survival proteins of the Bcl2 family.
  • a combination described herein comprises a Bcl-2 inhibitor.
  • the Bcl-2 inhibitor is venetoclax (also known as ABT-199): [282]
  • the Bcl-2 inhibitor is selected from the compounds described in WO 2013/110890 and WO 2015/011400.
  • the Bcl-2 inhibitor comprises navitoclax (ABT-263), ABT-737, BP1002, SPC2996, APG-1252, obatoclax mesylate (GX15-070MS), PNT2258, Zn-d5, BGB-11417, or oblimersen (G3139).
  • the Bcl-2 inhibitor is N-(4-hydroxyphenyl)-3-[6-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]- 1,3-benzodioxol-5-yl]-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide, compound A1: (compound A1).
  • the Bcl-2 inhibitor is (S)-5-(5-chloro-2-(3-(morpholinomethyl)- 1,2,3,4-tetrahydroisoquinoline-2-carbonyl)phenyl)-N-(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl)-N-(4- hydroxyphenyl)-1,2-dimethyl-1H-pyrrole-3-carboxamide), compound A2: (compound A2).
  • the antibody-drug conjugates or combinations disclosed herein are suitable for the treatment of cancer in vivo.
  • the combination can be used to inhibit the growth of cancerous tumors.
  • the combination can also be used in combination with one or more of: a standard of care treatment (e.g., for cancers or infectious disorders), a vaccine (e.g., a therapeutic cancer vaccine), a cell therapy, a hormone therapy (e.g., with anti-estrogens or anti-androgens), a radiation therapy, surgery, or any other therapeutic agent or modality, to treat a disorder herein.
  • a standard of care treatment e.g., for cancers or infectious disorders
  • a vaccine e.g., a therapeutic cancer vaccine
  • a cell therapy e.g., a hormone therapy (e.g., with anti-estrogens or anti-androgens)
  • a radiation therapy e.g., surgery, or any other therapeutic agent or modality
  • the combination can be administered together with an antigen of interest.
  • a combination disclosed herein can be administered in either order or simultaneously.
  • conjugates have favorable properties, for example properties that would make them easier to manufacture, easier to administer to patients, more efficacious, and/or potentially safer for patients.
  • One example is the determination of molecular size by size exclusion chromatography (SEC) wherein the amount of desired antibody species in a sample is determined relative to the amount of high molecular weight contaminants (e.g., dimer, multimer, or aggregated antibody) or low molecular weight contaminants (e.g., antibody fragments, degradation products, or individual antibody chains) present in the sample.
  • SEC size exclusion chromatography
  • a further example is the determination of the hydrophobicity by hydrophobic interaction chromatography (HIC) wherein the hydrophobicity of a sample is assessed relative to a set of standard antibodies of known properties.
  • HIC hydrophobic interaction chromatography
  • TLC Thin layer chromatography was conducted with 5 x 10 cm plates coated with Merck Type 60 F254 silica-gel.
  • Microwave Reactions [297] Microwave heating was performed with a CEM Discover® SP, or with an Anton Paar Monowave Microwave Reactor.
  • NMR [299] 1H-NMR measurements were performed on a Bruker Avance III 500 MHz spectrometer, a Bruker Avance III 400 MHz spectrometer, or a Bruker DPX-400 spectrometer using DMSO-d6 or CDCl3 as solvent.
  • 1H NMR data is in the form of delta values, given in part per million (ppm), using the residual peak of the solvent (2.50 ppm for DMSO-d6 and 7.26 ppm for CDCl3) as internal standard.
  • Splitting patterns are designated as: s (singlet), d (doublet), t (triplet), q (quartet), quint (quintet), sept (septet), m (multiplet), br s (broad singlet), dd (doublet of doublets), td (triplet of doublets), dt (doublet of triplets), ddd (doublet of doublet of doublets).
  • Acidic LCMS KINATEX XB-C18-100A, 2.6 ⁇ m, 50 mm*2.1 mm column at 40 °C, at a flow rate of 1 mL min-1 using 0.02% v/v aqueous formic acid (Solvent A) and 0.02% v/v formic acid in acetonitrile (Solvent B) with a gradient starting from 100% Solvent A and finishing at 100% Solvent B over various/certain duration of time.
  • Certain other compounds of the present invention were characterized HPLC-MS under specific named methods as follows. For all of these methods UV detection was by diode array detector at 230, 254, and 270 nm. Sample injection volume was 1 ⁇ L.
  • LCMS-V-B methods [309] Using an Agilent 1200 SL series instrument linked to an Agilent MSD 6140 single quadrupole with an ESI-APCI multimode source (Methods LCMS-V-B1 and LCMS-V-B2) or using an Agilent 1290 Infinity II series instrument connected to an Agilent TOF 6230 with an ESI-jet stream source (Method LCMS-V-B1); column: Thermo Accucore 2.6 ⁇ m, C18, 50 mm x 2.1 mm at 55 oC.
  • HPLC-V-A methods [316] These were performed on a Waters FractionLynx MS autopurification system, with a Gemini ® 5 ⁇ m C18(2), 100 mm ⁇ 20 mm i.d. column from Phenomenex, running at a flow rate of 20 cm 3 min -1 with UV diode array detection (210–400 nm) and mass-directed collection.
  • the mass spectrometer was a Waters Micromass ZQ2000 spectrometer, operating in positive or negative ion electrospray ionisation modes, with a molecular weight scan range of 150 to 1000.
  • HPLC-V-B methods [322] Performed on an AccQPrep HP125 (Teledyne ISCO) system, with a Gemini® NX 5 ⁇ m C18(2), 150 mm ⁇ 21.2 mm i.d. column from Phenomenex, running at a flow rate of 20 cm 3 min -1 with UV (214 and 254 nm) and ELS detection. [323] Method HPLC-V-B1 (pH 4): [324] Solvent A: water + 0.08% (v/v) formic acid; solvent B: acetonitrile + 0.08% (v/v) formic acid.
  • High-resolution MS High-resolution mass spectra were acquired on an Agilent 6230 time-of-flight mass spectrometer equipped with a Jet Stream electrospray ion source in positive ion mode. Injections of 0.5 ⁇ l were directed to the mass spectrometer at a flow rate 1.5 ml/min (5mM ammonium-formate in water and acetonitrile gradient program), using an Agilent 1290 Infinity HPLC system.
  • Jet Stream parameters drying gas (N2) flow and temperature: 8.0 l/min and 325 °C, respectively; nebulizer gas (N2) pressure: 30 psi; capillary voltage: 3000 V; sheath gas flow and temperature: 325 °C and 10.0 l/min; TOFMS parameters: fragmentor voltage: 100 V; skimmer potential: 60 V; OCT 1 RF Vpp:750 V. Full-scan mass spectra were acquired over the m/z range 105-1700 at an acquisition rate of 995.6 ms/spectrum and processed by Agilent MassHunter B.04.00 software.
  • Step B benzyl 6-[4-(ethoxycarbonyl)-1,5-dimethylpyrrol-2-yl]-7- ⁇ [(3R)-3-methyl-3,4- dihydro-1H-isoquinolin-2-yl]carbonyl ⁇ -3,4-dihydro-1H-isoquinoline-2-carboxylate [356] To a solution of Preparation I (3 g, 6.3 mmol, 1 eq.) and the product from Step A (1.27 g, 6.93 mmol, 1.1 eq.) in DMF (30 mL) was added DIPEA (3.13 mL, 18.89 mmol, 3 eq.) and PyBop (3.6 g, 6.93 mmol, 1.1 eq.) and the mixture stirred at rt for 12 h.
  • the mixture was diluted with water (120 mL), stirred for 15 mins and the resultant cream precipitate was collected by filtration, washing with water.
  • the filter cake was dissolved in DCM and washed with sat. aq. NaHCO3 solution and water, dried over MgSO4 and concentrated in vacuo to afford the title product (3.7 g, 6.11 mmol, 97%).
  • Step C ethyl 1,2-dimethyl-5-(7- ⁇ [(3R)-3-methyl-3,4-dihydro-1H-isoquinolin-2-yl]carbonyl ⁇ - 1,2,3,4-tetrahydroisoquinolin-6-yl)pyrrole-3-carboxylate 83 ME150999036v.1 132043-01120 [360] To a solution of the product from Step B (3.7 g, 6.11 mmol, 1 eq.) in MeOH (70 mL) and EtOH (20 mL) was added 10% Pd/C (100 mg).
  • Step D tert-butyl 6-[4-(ethoxycarbonyl)-1,5-dimethylpyrrol-2-yl]-7- ⁇ [(3R)-3-methyl-3,4- dihydro-1H-isoquinolin-2-yl]carbonyl ⁇ -3,4-dihydro-1H-isoquinoline-2-carboxylate [364] To a solution of the product from Step C (2.94 g, 6.23 mmol, 1 eq.) in THF (45 mL) and water (6 mL) was added bis(tert-butyl) dicarbonate (1.43 g, 6.55 mmol, 1.05 eq.) followed by TEA (1.73 mL, 12.47 mmol, 2 eq.) and the mixture stirred at rt for 12 h.
  • Step E 5-[2-(tert-butoxycarbonyl)-7- ⁇ [(3R)-3-methyl-3,4-dihydro-1H-isoquinolin-2- yl]carbonyl ⁇ -3,4-dihydro-1H-isoquinolin-6-yl]-1,2-dimethylpyrrole-3-carboxylic acid
  • LiOH.H2O 995 mg, 23.72 mmol, 4 eq.
  • MeOH 40 mL
  • water 20 mL
  • Step B 4-( ⁇ 4-[(tert-butyldimethylsilyl)oxy]phenyl ⁇ amino)-1,5-dimethylpyrrole-2-carbonitrile
  • Step A Using General procedure 1a and the product from Step A (6.1 g, 30.65 mmol, 1 eq.) as the appropriate aryl bromide and Preparation IIIa (7.19 g, 32.18 mmol, 1.05 eq.) as the appropriate aniline afforded the title product (8.2 g, 24 mmol, 78%).
  • Step C tert-butyl 6-[4-( ⁇ 4-[(tert-butyldimethylsilyl)oxy]phenyl ⁇ (5-cyano-1,2-dimethylpyrrol- 3-yl)carbamoyl)-1,5-dimethylpyrrol-2-yl]-7- ⁇ [(3R)-3-methyl-3,4-dihydro-1H-isoquinolin-2- yl]carbonyl ⁇ -3,4-dihydro-1H-isoquinoline-2-carboxylate [384] The title compound was prepared according to General procedure 2a using Preparation IIa (2.5 g, 4.6 mmol, 1 eq.) as the appropriate acid and the product from Step B (3.14 g, 9.2 mmol, 2 eq.) as the appropriate aniline.
  • Preparation IIa 2.5 g, 4.6 mmol, 1 eq.
  • Step D N-(5-cyano-1,2-dimethylpyrrol-3-yl)-N-(4-hydroxyphenyl)-1,2-dimethyl-5-(7- ⁇ [(3R)- 3-methyl-3,4-dihydro-1H-isoquinolin-2-yl]carbonyl ⁇ -1,2,3,4-tetrahydroisoquinolin-6-yl)pyrrole-3- carboxamide hydrochloride [387] A solution of the product from Step C (2.8 g, 3.23 mmol, 1 eq.) in MeOH (5 mL) was treated with 3 M HCl in MeOH (10 mL
  • Step B ⁇ 4-[2-(morpholin-4-yl)ethoxy]phenyl ⁇ acetic acid
  • Step B 1,2-dimethyl-5-(7- ⁇ [(3R)-3-methyl-3,4-dihydro-1H-isoquinolin-2-yl]carbonyl ⁇ -2-(2- ⁇ 4-[2-(morpholin-4-yl)ethoxy]phenyl ⁇ acetyl)-3,4-dihydro-1H-isoquinolin-6-yl)pyrrole-3-carboxylic acid
  • Preparation Va (1 g, 3.77 mmol, 1 eq) in anhydrous DCM (10 mL) under N2
  • 2 M oxalyl chloride solution in DCM (0.43 mL, 4.52 mmol, 1.2 eq.
  • Step A of Preparation IIa After stirring for 10 mins the product from Step A of Preparation IIa (18.6 g, 101 mmol, 1.05 eq.) was added and the mixture was stirred at rt for 1h then it was poured into water (1500 mL) and the precipitates were filtered out, washed with water.
  • This crude intermediate was dissolved in methanol (250 mL) and water (25 mL) then NaOH (28 g, 700 mmol, 7.3 eq.) was added and mixture was stirred at reflux temperature for 18 h. Evaporated at reduced pressure, then the pH was adjusted to 6 by the addition of cc. aq. HCl.
  • Step B 5-[2-(9H-fluoren-9-ylmethoxycarbonyl)-7-[(3R)-3-methyl-3,4-dihydro-1H- isoquinoline-2-carbonyl]-3,4-dihydro-1H-isoquinolin-6-yl]-1,2-dimethyl-pyrrole-3-carboxylic acid [413] To the biphasic mixture of the product from Step A (13.96 g, 28.33 mmol, 1 eq.) dissolved in dioxane (160 mL) and NaHCO 3 (5.47 g, 65.2 mmol, 2.3 eq.) dissolved in water (160 mL) 9H-fluoren- 9-ylmethyl carbonochloridate (8.06 g, 31.2 mmol, 1.1
  • Step C 9H-fluoren-9-ylmethyl 6-[4-[[4-[tert-butyl(dimethyl)silyl]oxyphenyl]-(5-cyano-1,2- dimethyl-pyrrol-3-yl)carbamoyl]-1,5-dimethyl-pyrrol-2-yl]-7-[(3R)-3-methyl-3,4-dihydro-1H- isoquinoline-2-carbonyl]-3,4-dihydro-1H-isoquinoline-2-carboxylate [417] Using General procedure 2b and the product from Step B (10.67 g, 16.03 mmol, 1 eq.) as the appropriate acid, oxalyl dichloride (2.17 mL, 25.6 mmol, 1.6 eq.) and the product
  • Step D N-[4-[tert-butyl(dimethyl)silyl]oxyphenyl]-N-(5-cyano-1,2-dimethyl-pyrrol-3-yl)-1,2- dimethyl-5-[7-[(3R)-3-methyl-3,4-dihydro-1H-isoquinoline-2-carbonyl]-1,2,3,4- tetrahydroisoquinolin-6-yl]pyrrole-3-carboxamide [421] To a solution of the product from Step C (8.12 g, 8.21 mmol, 1 eq.) in DCM (41 mL) morpholine (41 mL, 475 mmol, 58 eq.) was added then the mixture was stirred at rt for 18 h.
  • reaction mixture is stirred at 0°C for 1 hour. Then, the reaction mixture is hydrolysed by addition of 420 mL of ice-cold water, diluted with ethyl acetate, and successively washed with 0.1M aqueous hydrochloric acid (HCl) solution, saturated aqueous LiCl solution and then brine. The organic phase is dried over MgSO4, filtered, concentrated to dryness and purified by chromatography over silica gel (petroleum ether/AcOEt gradient).
  • HCl hydrochloric acid
  • Step 2 Ethyl 5-(5-chloro-2-formylphenyl)-1,2-dimethyl-1H-pyrrole-3-carboxylate [429] To a solution of the compound obtained in Step 1 (10.5 g, 62,8 mmol) in 65 mL of N,N- dimethylacetamide there are successively added 2-bromo-4-chlorobenzaldehyde (15.2 g, 69 mmol), potassium acetate (12.3 g, 125.6 mmol) and then the reaction mixture is stirred under argon for 20 minutes. There is added PdCl2(PPh3)2 (2.2 g, 3.14 mmol).
  • the reaction mixture is heated at 130°C overnight, allowed to cool down to room temperature and it is diluted with dichloromethane.
  • Animal charcoal is added (20g), the suspension is stirred at room temperature for 1 hour and filtered.
  • the organic phase is washed with water, dried over MgSO 4 and concentrated to dryness.
  • the crude product thereby obtained is purified by chromatography over silica gel (petroleum ether/AcOEt gradient). The title product is obtained in the form of a solid.
  • Step 3 4-Chloro-2-[4-(ethoxycarbonyl)-1,5-dimethyl-1H-pyrrol-2-yl]benzoic acid
  • a solution is prepared containing the compound obtained in Step 2 (12.8 g, 42 mmol) and 2- methyl-2-butene (35.7 mL, 336 mmol) in a mixture containing 20 mL of acetone and 20 mL of tetrahydrofuran. There are added, dropwise, 200 mL of an aqueous solution containing a mixture of sodium chlorite (NaClO2, 13.3 g, 147 mmol) and sodium hydrogen phosphate (NaHPO4, 14.5 g, 105 mmol).
  • NaClO2 sodium chlorite
  • NaHPO4 sodium hydrogen phosphate
  • the reaction mixture is vigorously stirred at room temperature for 7 hours and concentrated to remove the acetone.
  • Ethyl acetate is added, the organic phase is washed with water, dried over MgSO 4 and then concentrated to dryness. The residue is then taken up in a minimum of ethyl ether.
  • the solid then obtained is filtered off, washed with ether and then dried in vacuo at 40°C overnight.
  • the title product is obtained in the form of a solid, which is subsequently used without being otherwise purified.
  • Step 4 ⁇ (3S)-2-[(4-Methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinolin-3-yl ⁇ methyl 4- methylbenzenesulphonate 93 ME150999036v.1 132043-01120 [435] To a solution of commercially available [(3S)-1,2,3,4-tetrahydroisoquinolin-3-yl]methanol (30.2 g, 185 mmol) in 750 mL of dichloromethane there are successively added tosyl chloride (91.7 g, ,481 mmol) and then, dropwise, N,N,N-triethylamine (122 mL, 740 mmol).
  • the reaction mixture is stirred at room temperature for 20 hours, diluted with dichloromethane, washed successively with 1M HCl solution, saturated aqueous NaHCO3 solution and then brine until neutral.
  • the organic phase is then dried over MgSO4, filtered and concentrated to dryness.
  • the solid obtained is dissolved in a minimum volume of dichloromethane and then cyclohexane is added until a precipitate is formed. This precipitate is then filtered off and washed with cyclohexane. After drying, the title product is obtained in the form of a white crystalline powder.
  • Step 5 (3S)-3-(iodomethyl)-2-(p-tolylsulfonyl)-3,4-dihydro-1H-isoquinoline [438] To a suspension of the compound obtained in Step 4 (4 g, 8.5 mmol) in acetonitrile (10 mL) is added sodium iodide (1.4 g, 9.3 mmol). The reaction mixture is heated under microwave irradiations (100W for 6h), cooled to room temperature. The suspension is filtered. The solid is washed with dichloromethane. The filtrate and the washings are pooled together and concentrated to dryness.
  • Step 6 diethyl 2-[[(3R)-2-(p-tolylsulfonyl)-3,4-dihydro-1H-isoquinolin-3- yl]methyl]propanedioate
  • To a suspension of sodium hydride (442 mg, 11 mmol) in THF (8 mL) is added dropwise diethyl malonate (1.5 mL, 10 mmol) at room temperature. After 15 minutes, a solution of the compound obtained in Step 5 (4.2 g, 10 mmol) in THF (10 mL) is added dropwise. After 20 minutes at room temperature, the reaction mixture is added in microwave reactor (100°C-275W) during 19h.
  • the reaction mixture is extracted with ethyl acetate twice. The organic phase is washed with brine, dried over MgSO4 and concentrated to dryness.
  • the crude material is diluted with DMSO (10 mL) and a solution of sodium chloride (0.25 g, 4.2 mmol) in water (1 mL). The reaction mixture is heated to 140°C for 1h, then cooled to room temperature and diluted with ethyl acetate. The organic layer is separated, dried over MgSO4 and concentrated to dryness, affording the desired compound.
  • Step 8 N,N-dimethyl-3-[(3R)-2-(p-tolylsulfonyl)-3,4-dihydro-1H-isoquinolin-3- yl]propenamide
  • N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride 5.4 g, 28 mmol
  • 1- hydroxybenzotriazole hydrate 3.8 g, 28 mmol
  • N,N-diisopropylethylamine (6.9 mL, 41 mmol)
  • 2 M dimethylamine in THF (20.8 mL).
  • reaction mixture is stirred overnight at room temperature, diluted with dichloromethane, washed successively with water, aqueous 1M HCl solution and brine.
  • organic phase is dried over MgSO 4 , concentrated to dryness and purified by chromatography over silica gel using cyclohexane and ethyl acetate as eluants, affording the desired compound.
  • Step 9 N,N-dimethyl-3-[(3R)-2-(p-tolylsulfonyl)-3,4-dihydro-1H-isoquinolin-3-yl]propan-1- amine
  • Step 8 To a solution of Step 8 (6.6 g, 17.1 mmol) in anhydrous THF (70 mL) is added dropwise under inert atmosphere a solution of 1M borane tetrahydrofuran complex in tetrahydrofuran (69 mL, 69 mmol). The reaction mixture is heated to 70°C overnight and cooled to room temperature. The reaction mixture is diluted with dichloromethane and water.
  • the organic phase is washed successively with an aqueous 1M HCl solution, a saturated aqueous NaHCO 3 solution, brine, and dried over MgSO 4 and concentrated.
  • the crude mixture is purified by chromatography over silica gel using dichloromethane and NH 3 2M in ethanol as eluants, affording the desired compound.
  • Step 10 N,N-dimethyl-3-[(3R)-1,2,3,4-tetrahydroisoquinolin-3-yl]propan-1-amine [454] To a solution of naphthalene (13 g, 103 mmol) in anhydrous THF (35 mL) under argon atmosphere is added sodium (2.4 g, 103 mmol) portionwise. After stirring 1h at room temperature, the reaction mixture is cooled to -78°C and a solution of Step 9 (4.8 g, 12.2 mmol) in THF (35 mL) is added dropwise.
  • reaction mixture After 3 h of stirring at -78°C, the reaction mixture is allowed to warm to 0°C and a solution of saturated aqueous ammonium chloride is carefully added (3 mL). The reaction mixture is allowed to warm to room temperature and is evaporated to dryness. The crude mixture is purified by chromatography over silica gel using dichloromethane and NH32M in ethanol as eluants, affording the desired compound.
  • Step 11 ethyl 5-[5-chloro-2-[(3R)-3-[3-(dimethylamino)propyl]-3,4-dihydro-1H- isoquinoline-2-carbonyl]phenyl]-1,2-dimethyl-pyrrole-3-carboxylate [457] To a solution of the compound obtained in Step 3 (15.5 g, 48 mmol) in dichloromethane (3L) are added N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (9.2 g, 48 mmol), 1- hydroxybenzotriazole hydrate (7.4 g, 48 mmol), the compound obtained in Step 10 (10 g, 48 mmol) and N-ethyl-N-isopropyl-propan-2-amine (28 mL, 160 mmol).
  • reaction mixture is stirred for 3h at room temperature, diluted with dichloromethane, washed with water, concentrated to dryness and purified by chromatography over silica gel using dichloromethane and NH 3 2M in ethanol as eluants, affording the desired compound.
  • Step 12 5-[5-chloro-2-[(3R)-3-[3-(dimethylamino)propyl]-3,4-dihydro-1H-isoquinoline-2- carbonyl]phenyl]-1,2-dimethyl-pyrrole-3-carboxylic acid [461] To a solution of the compound obtained in Step 11 (400 mg, 0.76 mmol) in methanol (2 mL) is added a solution of lithium hydroxide monohydrate (112 mg, 2.6 mmol) in water (2 mL). The reaction mixture is heated at reflux overnight then partially concentrated.
  • Step 13 N-[4-[tert-butyl(dimethyl)silyl]oxyphenyl]-5-[5-chloro-2-[(3R)-3-[3- (dimethylamino)propyl]-3,4-dihydro-1H-isoquinoline-2-carbonyl]phenyl]-N-(5-cyano-1,2-dimethyl- pyrrol-3-yl)-1,2-dimethyl-pyrrole-3-carboxamide [464] To a solution of the compound obtained in Step 12 (480 mg, 0.97 mmol) in 1,2- dichloroethane (60 mL) is added 1-chloro-N,N,2-trimethyl-prop-1-en-1-amine (170 ⁇ L, 1.3 mmol).
  • Step 14 5-[5-chloro-2-[(3R)-3-[3-(dimethylamino)propyl]-3,4-dihydro-1H-isoquinoline-2- carbonyl]phenyl]-N-(5-cyano-1,2-dimethyl-pyrrol-3-yl)-N-(4-hydroxyphenyl)-1,2-dimethyl-pyrrole-3- carboxamide [469] To a solution of the compound obtained in Step 13 (0.5 g, 0.61 mmol) in THF (20 mL) is added at room temperature a solution of 1M tetra-n-butylammonium fluoride in THF (1 mL, 1 mmol).
  • the reaction mixture is stirred for 4h at room temperature and diluted with dichloromethane and a saturated aqueous solution of NaHCO3.
  • the organic phase is separated, washed successively with water, brine, then dried over MgSO4 and concentrated.
  • the crude is purified by chromatography over silica gel using dichloromethane and NH32M methanol as eluants, affording the desired compound.
  • Step B 9H-fluoren-9-ylmethyl 6-[4-[[4-[tert-butyl(dimethyl)silyl]oxyphenyl]-[1- (difluoromethyl)pyrazol-4-yl]carbamoyl]-1,5-dimethyl-pyrrol-2-yl]-7-[(3R)-3-methyl-3,4-dihydro- 1H-isoquinoline-2-carbonyl]-3,4-dihydro-1H-isoquinoline-2-carboxylate [479] Using the procedure described in Preparation VIIb Step C and the product from Step A (1.16 g, 3.42 mmol) as the appropriate aniline instead of the product of Preparation IVa Step B afforded the title product (2.18 g, 77 %).
  • Step C N-[4-[tert-butyl(dimethyl)silyl]oxyphenyl]-N-[1-(difluoromethyl)pyrazol-4-yl]-1,2- dimethyl-5-[7-[(3R)-3-methyl-3,4-dihydro-1H-isoquinoline-2-carbonyl]-1,2,3,4- tetrahydroisoquinolin-6-yl]pyrrole-3-carboxamide [483] Using the procedure described in Preparation VIIb Step D and the product from Step B (2.18 g, 2.21 mmol) instead of the product from Preparation VIIb Step C afforded the title product (0.83 g, 49 %).
  • Step D N-[4-[tert-butyl(dimethyl)silyl]oxyphenyl]-N-[1-(difluoromethyl)pyrazol-4-yl]-1,2- dimethyl-5-[7-[(3R)-3-methyl-3,4-dihydro-1H-isoquinoline-2-carbonyl]-2-[2-[4-(2- morpholinoethoxy)phenyl]acetyl]-3,4-dihydro-1H-isoquinolin-6-yl]pyrrole-3-carboxamide [487] Using modified General procedure 4a (using TBTU instead of HATU and omitting preparative HPLC flash chromatography), starting from the product of Preparation Va (283 mg, 1.2 eq.) as the appropriate carboxylic acid and the product from Step C (680 mg, 0.889 mmol) as the appropriate amine, afforded the title compound
  • Step E N-[1-(difluoromethyl)pyrazol-4-yl]-5-[2-[2-[4-(2-morpholinoethoxy) phenyl]acetyl]- 7-[(3R)-3-methyl-3,4-dihydro-1H-isoquinoline-2-carbonyl]-3,4-dihydro-1H-isoquinolin-6-yl]-N-(4- hydroxyphenyl)-1,2-dimethyl-pyrrole-3-carboxamide [491] To the product from Step D (100 mg, 0.099 mmol) dissolved in acetonitrile (150 mL/mmol) 2M aq.
  • Step B N-(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl)-N-(4-hydroxyphenyl)-1,2-dimethyl-5- ⁇ 7- [(3R)-3-methyl-1,2,3,4-tetrahydroisoquinoline-2-carbonyl]-2-(2- ⁇ 4-[2-(morpholin-4- yl)ethoxy]phenyl ⁇ acetyl)-1,2,3,4-tetrahydroisoquinolin-6-yl ⁇ -1H-pyrrole-3-carboxamide [498] According the procedure described in Step E of the Preparation of P3, the product from Step A (390 mg, 0.385 mmol) was treated with 2M aq.
  • Linker-payloads P1-L12-P2, P1-L12-P3, P1-L12-P4, P1-L17-P2, P1-L17-P3, P1-L17-P4, and P1-L19-P2 were prepared according to the methods described in International PCT PCT/US2023/022990, which is incorporated herein by reference, and as described herein. Below is a table showing structures of these Linker-payloads.
  • NMR data were acquired at a temperature of 298K on a Bruker Avance NMR spectrometer equipped with a 5 mm BBFO CryoProbe with z-gradient operating at a frequency of 400.13 MHz for 1 H, 376.50 MHz for 19 F, 100.61 MHz for 13 C.
  • Chemical shifts for the 1 H and 13 C spectra were referenced by setting internal tetramethylsilane (TMS) to 0 ppm.
  • TFA method solvent: A water + 0.05 % TFA, B acetonitrile + 0.05 % TFA, gradient from 5 to 100% B in 15 to 30 CV b .
  • NH4HCO3 method solvent: A water + 0.02 M NH4HCO3, B acetonitrile/water 80/20 + 0.02 M NH4HCO3, gradient from 5 to 100 % B in 15 to 30 CV c .
  • Neutral method solvent: A water, B acetonitrile, gradient from 5 to 100% B in 15 to 30 CV d .
  • Formic Acid method solvent: A water + 0.05 % Formic Acid, B acetonitrile + 0.05 % Formic Acid, gradient from 5 to 100% B in 15 to 30 CV Gradient variations of methods a.-d. were employed as appropriate. All the fractions containing the pure compound were combined and directly freeze-dried to afford the compound as an amorphous powder.
  • Step 1 Synthesis of (9H-fluoren-9-yl)methyl (5-amino-2-(((tert- butyldiphenylsilyl)oxy)methyl)benzyl)(methyl)carbamate
  • Step 1 Synthesis of 2-(hydroxymethyl)-N-methyl-5-nitrobenzamide
  • MeNH2 3.00 kg, 29.94 mol, 600 mL, 31.0% purity
  • Step 2 Synthesis of (2-((methylamino)methyl)-4-nitrophenyl)methanol
  • a solution of 2-(hydroxymethyl)-N-methyl-5-nitrobenzamide (560 g, 2.66 mol) in THF (5000 mL) was cooled to 0 °C, then BH3-Me2S (506 g, 6.66 mol) (2.0 M in THF) was added drop wise for 60 min and the mixture was heated to 70 °C for 5 h.
  • LCMS showed the starting material was consumed.
  • 4M HCl (1200 mL) in Methanol was added to the reaction mixture at 0 °C and heated at 65 °C for 8 h.
  • Step 5 Synthesis of (9H-fluoren-9-yl)methyl (5-amino-2-(((tert- butyldiphenylsilyl)oxy)methyl)benzyl)(methyl)carbamate
  • reaction solution was dripped into 4L H2O with stirring over ca.30 minutes at which time the ppt was filtered, rinsed with H2O (1L) and air dried under vacuum.
  • the wet cake was dissolved in 15% iPrOH/EtOAc (ca.
  • the reaction was kept stirring in the ice bath for 1 hr. The reaction was slowly warmed up to RT and stirred at RT for 1 hour. The volatiles were removed in-vacuo. The product was retaken in dicholoroethane (210 mL). The resulting solution was removed in-vacuo. The process was repeated 2 more times (3 times in total) to obtain a dark yellow solid. The solid was suspended in diethyl ether (300 mL). It was sonicated for 10 min then decanted. The process was repeated 2 more times. The solid was collected by vacuum filtration. After air drying overnight under vacuum, the solid was dissolved in MeOH (100 mL) and this solution was added dropwise over ca.
  • the solution was purified by ISCO C18 RP-HPLC. After lyophilization, the residue (630 mg) was dissolved in DMF (2 mL) and 2M dimethyl amine in MeOH (2 mL, ca. 4000 ⁇ mol) was added. After standing two hours, 1.0 M tetrabutyl ammonium fluoride (0.20 mL, 200 ⁇ mol) was added. After 60 minutes, the solution was diluted in DMSO and purified by C18 ISCO RP-HPLC.
  • Acetic acid (0.036 mL, 0.627 mmol) and DIPEA (1.096 mL, 6.27 mmol) were added.
  • the reaction atmosphere was switched to hydrogen (treat reaction chamber with vacuum and refill with nitrogen, repeat 4x.
  • Then treat 130 ME150999036v.1 132043-01120 reaction chamber with vacuum and refill with hydrogen, repeat 4x).
  • the reaction was stirred at RT for 16 hr.
  • the reaction solution was filtered through a celite pad. The pad was then washed with EtOAc (20 mL).
  • EtOAc (20 mL).
  • the combined organic layer was removed under reduced pressure.
  • the product was then dried under high vacuum for 30 min.
  • the mixture was stirred at rt for 4 h, and was concentrated to ca. 400 mL and was washed with 1:1 brine / water (150 mL). The aqueous was extracted with CH2Cl2 (20 mL). The washing and back extraction procedure was repeated three more times. The combined organic phase was washed with satd. NaHCO3 (50 mLx2), brine, dried over anh.
  • the bottom oily material was washed with fresh 1:1 ether / heptane (2 X50 mL) and dried in vacuum to give an oil (7.718 g) as HOAc salt of the title compound.
  • the material was dissolved with 1/9 TFE / CH2Cl2 (250 mL) and was washed with satd. NaHCO3 (2X 100 mL) until the aqueous was basic.
  • the combined organic phase was washed with brine (75 mL).
  • the aqueous was extracted with 1/9 TFE / CH2Cl2 (20 mL) twice.
  • the combined organic phase was dried over anh.
  • the mixture was stirred at rt/20 h, The volatile was removed via rotary evaporation.
  • the resulting oil was diluted with DMSO (6 mL) and was purified by RP-HPLC (C18, ACN/water with 0.1% NH4OH).
  • the desired product-containing fractions (total ca.112 mL) were diluted with EtOAc (75 mL) and acidified with 1 N HCl (3.5 mL).
  • the organic phase was separated.
  • the aqueous was extracted with EtOAc (2X30 mL).
  • the combined EtOAc extract was mixed with heptane (20 mL) and the mixture was washed with brine (2X15 mL), dried over anh.
  • the resin was quickly washed with 50 column volumes PBS on a vacuum manifold in multiple additions. The resin was then resuspended in an equal volume PBS containing 250 nM CuCl2. Reformation of antibody interchain disulfides was monitored by taking time points. At each time point, 25 ⁇ L of resin slurry was removed, 1 ⁇ L of 20 mM MC-valcit-MMAE was added, and the tube flicked several times. The resin was spun down, supernatant removed, and then eluted with 50 ⁇ L Antibody elution buffer (Thermo).
  • the resin was pelleted and the supernatant analyzed by reverse phase chromatography using an Agilent PLRP-S 4000A 5um, 4.6x50mm column (Buffer A is water, 174 ME150999036v.1 132043-01120 0.1% TFA, Buffer B Acetonitrile, 0.1% TFA, column held at 80°C, Flowrate 1.5 ml/min; Gradient from 30-45% B, 0-5 min, 100%B 5.01-6.5 min, re-equilibrate to 30% B).
  • the resin was washed with 10 column volumes PBS and the resin was resuspended in an equal volume PBS and between 8-12 equivalents of the indicated linker-payload (20 mM) in DMSO was added, with a final concentration of 10% DMSO in reaction and then incubated at room temperature for 3 hours. The resin was then washed with 50 column volumes PBS to remove excess linker-payload.
  • the ADC was eluted from the protein A resin with Antibody elution buffer. The ADC was then buffer exchanged into PBS or other suitable buffer.
  • DAR drug-to-antibody ratio
  • LC/MS liquid chromatography-mass spectrometry
  • mobile phase A was purified MS grade water (Honeywell, LC015-l)
  • mobile phase B was MS grade 80% Isopropanol (Honeywell LC323-l): 20% acetonitrile (Honeywell, LC015-l), LC323-1), supplemented with 1 % of formic acid (FA) (Thermo Scientific, 85178).
  • the column temperature was set at 80°C.
  • a general MS method was optimized for all ADCs synthesized.
  • the column used for analysis was an Agilent PLRP-S 4000 A; 2.lx150mm, 8um (Agilent, PL1912-3803). Flowrate used was 0.3 ml/min.
  • the gradient used was 0-0.75 minute 95%A, 0.76 -1.9 minute 75%A, 1.91-11.0 minute 50%A, 11.01-11.5010%A, 11.51-13.50 minute 95%A, 13.51-18 minute 95%A on an Acuity Bio H-Class Quaternary UPLC (Waters).
  • MS system was Xevo G2-XS QToF ESI mass spectrometer (Waters) and data acquired from 1.5-11 minutes and masses were analyzed between 15000-80000 daltons.
  • DAR was determined from the deconvoluted spectra or UV chromatogram by summing the integrated MS (total ion current) or UV (280 nm) peak area of unconjugated and conjugated given species (mAb or associated fragment), weighted by multiplying each area by the number of drug attached. The summed, weighted areas were divided by the sum of total area and the results produced a final average DAR value for the full ADC.
  • the analysis was performed on analytical column Superdex 200 Increase 5/150 GL (GE Healthcare, 28990945) in isocratic conditions 100% PBS pH 7.2 ((Hyclone SH30028.03)), flow 0.45 ml/min for 8 minutes.
  • the % aggregate fraction of the ADC sample was quantified based on the peak area absorbance at 280 nm. Calculation was based on the ratio between the high molecular weight eluent at 280 nm divided by the sum of peak area absorbance at the same wavelength of the high molecular weight and monomeric eluents multiplied by 100%.
  • the peptide was dissolved at 50 mM in 90% dDMSO.
  • a CD74 Fab fragment (VHmil x Vk1aNQ) was dissolved at 0.52 mM (24.8 mg/ml) in 0.02 M phosphate buffer pH7.2, 0.2 M sucrose, 5% glycerol, 0.01% CHAPS and 0.001M TCEP HCl.
  • 6 ⁇ l of peptide solution were added to 150 ⁇ l of Fab solution resulting in a 4x fold molar excess of peptide over the Fab.
  • the complex solution was incubated at room temperature for 1h before setting up the crystallization experiment.
  • X-ray diffraction data were collected at the Swiss Light Source, beamline X10SA, with an Eiger pixel detector (Dectris) using a monochromatic X-ray beam of 1.0000 ⁇ wavelength.
  • Raw diffraction data were processed using the autoPROC software pipeline (Global Phasing Ltd, Cambridge UK). The crystal diffracted to 1.6 ⁇ resolution and belonged to space group H3. Structure determination, refinement and analysis [599] The structure of the CD74 peptide/Fab complex was solved by molecular replacement with PHASER (McCoy et al., J. Appl. Crystallogr.
  • the CD74 dual antibody drug conjugates were tested against three endogenous AML cancer cell lines: NOMO-1: DSMZ No. ACC 542 cultured in RPMI-1640 + 10% FBS EOL-1: DSMZ No. ACC 386 cultured in RPMI-1640 + 10% FBS MONO-MAC-1: DSMZ No. ACC 252 cultured RPMI-1640 + 10% FBS Inhibition of cell proliferation and survival [606] The ability of the CD74 dual antibody drug conjugate to inhibit cell proliferation as a monotherapy was assessed using the Promega CellTiter-Glo ® proliferation assay. [607] Cell lines were cultured in media that is optimal for their growth at 5% CO2, 37°C in a tissue culture incubator.
  • the cells Prior to seeding for the proliferation assay, the cells were split at least 2 days before the assay to ensure optimal growth density. On the day of seeding, suspension cells were harvested. Cell viability and cell density were determined using a cell counter (Vi-Cell XR Cell Viability Analyzer, Beckman Coulter). Cells with higher than 85% viability were seeded in white clear bottom 384-well TC treated plates (Corning cat. # 3765). Cells were seeded at a density of 1,000 cells per well in 45 ⁇ L of standard growth media. The primary AML PDX ex vivo samples were seeded at a density of 10,000 cells per well in 45 ⁇ L of standard growth media.
  • Plates were incubated at room temperature for 10 minutes to stabilize the luminescent signals prior to reading.
  • the luminescent signal for all plates was measured using a luminescence reader (PHERAstar FSX Plate Reader, BMG Labtech). This signal represents the cell viability of untreated cells on the day of dosing, or day 0. [609] Plates were incubated at 5% CO2, 37°C for 5 days in a tissue culture incubator, after which cell viability was assessed through the addition of 25 ⁇ L of CellTiter Glo® (Promega, cat# G7573). Plates were incubated at room temperature for 10 minutes to stabilize luminescent signals prior to reading using a luminescence reader (PHERAstar FSX Plate Reader, BMG Labtech).
  • EOL1 cells expressing luciferase were cultured at 37 °C (atmosphere of 5% CO2) in RPMI 1640 media supplemented with 2mM Glutamine and 10% Fetal Bovine Serum (FBS). The cell density was maintained between 0.3-1.5 million cells/mL throughout the expansion.
  • EOL1-Luc xenografts cells were harvested and re-suspended in phosphate buffered saline. A total of 1 x 10 6 EOL1-Luc cells were injected intravenously into female NSG mice (The Jackson Laboratory, USA) in a volume of 200 ⁇ L.
  • %T/C values were calculated according to the following formula: (( ⁇ Photons/sec in experimental group) / ( ⁇ Photons/sec in vehicle control group))*100.
  • Tumor regression values were calculated according to the following formula: (( ⁇ Photons/sec in experimental group) / (Photons/sec in experimental group at start of treatment))*100. Results are presented in Table 7 as mean ⁇ SEM.
  • Tumor volume data were analyzed for statistical significance relative to the untreated control. The statistical analysis is presented in Table 7. Unpaired two-tailed T-tests were used to make comparisons between groups. Table 7.
  • the CD74 VHmil x VK1aNQ-P1-L12-P4 ADC reduced the leukemic burden in an EOL1- Luc disseminated tumor model. Regressions were observed early after the first dose of ADC, at dose levels as low as 2.5 mpk. Relapse was observed by day 11 post first dose. A second dose of the CD74 VHmil x VK1aNQ-P1-L12-P4 ADC was administered on Day 15, however the effect of this dose what rather limited, as tumor burden continued to increase.

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Abstract

L'invention concerne des conjugués anticorps anti-CD74-médicament. Les conjugués anticorps-médicament comprennent un anticorps anti-CD74 ou un fragment de liaison à l'antigène de celui-ci lié de manière covalente à deux charges utiles antinéoplasiques par l'intermédiaire d'un lieur double. L'invention concerne en outre des méthodes et des compositions destinées à être utilisées dans le traitement de cancers par administration des conjugués anticorps-médicament décrits ici.
PCT/US2024/056875 2023-11-22 2024-11-21 Conjugués anticorps anti-cd74-médicament et leurs procédés d'utilisation Pending WO2025111450A1 (fr)

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Citations (62)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2779780A (en) 1955-03-01 1957-01-29 Du Pont 1, 4-diamino-2, 3-dicyano-1, 4-bis (substituted mercapto) butadienes and their preparation
US4816567A (en) 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
WO2000035436A2 (fr) 1998-12-16 2000-06-22 Warner-Lambert Company Traitement de l'arthrite a l'aide d'inhibiteurs de la mek
WO2001051056A1 (fr) 2000-01-10 2001-07-19 Astrazeneca Ab Preparation de fulvestrant
WO2002006213A2 (fr) 2000-07-19 2002-01-24 Warner-Lambert Company Esters oxygenes d'acides 4-iodophenylamino benzhydroxamiques
WO2003076424A1 (fr) 2002-03-08 2003-09-18 Eisai Co. Ltd. Composes macrocycliques utiles comme produits pharmaceutiques
WO2003077914A1 (fr) 2002-03-13 2003-09-25 Array Biopharma, Inc Utilisation de derives de benzimidazole alkyles n3 en tant qu'inhibiteurs de mek
US6703199B1 (en) 1997-06-12 2004-03-09 Research Corporation Technologies, Inc. Artificial antibody polypeptides
WO2005121142A1 (fr) 2004-06-11 2005-12-22 Japan Tobacco Inc. Dérivés de 5-amino-2,4,7-trioxo-3,4,7,8-tétrahydro-2h-pyrido’2,3-d! pyrimidine et composés apparentés pour le traitement du cancer
WO2006034488A2 (fr) 2004-09-23 2006-03-30 Genentech, Inc. Anticorps et conjugués produits avec de la cystéine
WO2007004415A1 (fr) 2005-07-01 2007-01-11 Murata Manufacturing Co., Ltd. Substrat céramique à couches multiples, procédé pour le fabriquer et feuille verte composite pour la fabrication dudit substrat
WO2008016893A1 (fr) 2006-08-02 2008-02-07 Novartis Ag Composés organiques
WO2008024725A1 (fr) 2006-08-21 2008-02-28 Genentech, Inc. Composés aza-benzofuranyle et leurs procédés d'utilisation
WO2009085983A1 (fr) 2007-12-19 2009-07-09 Genentech, Inc. 5-anilinoimidazopyridines et procédés d'utilisation
US7612114B2 (en) 2002-12-26 2009-11-03 Eisai R&D Management Co., Ltd. Selective estrogen receptor modulator
WO2010020675A1 (fr) 2008-08-22 2010-02-25 Novartis Ag Composés de pyrrolopyrimidine et leurs utilisations
WO2010080478A1 (fr) 2008-12-19 2010-07-15 Genentech, Inc. Composés et méthodes d'utilisation
WO2010080503A1 (fr) 2008-12-19 2010-07-15 Genentech, Inc. Composés hétérocycliques et leurs procédés d'utilisation
WO2011005481A1 (fr) 2009-06-22 2011-01-13 Medimmune, Llc Régions fc de synthèse pour une conjugaison spécifique à un site
WO2011076786A1 (fr) 2009-12-22 2011-06-30 Novartis Ag Isoquinolinones et quinazolinones substituées
WO2011156518A2 (fr) 2010-06-10 2011-12-15 Aragon Pharmaceuticals, Inc. Modulateur du récepteur oestrogénique et utilisation de ces derniers
WO2011159769A2 (fr) 2010-06-17 2011-12-22 Aragon Pharmaceuticals, Inc. Modulateurs de récepteur d'œstrogène d'indane et utilisations de ceux-ci
WO2012037410A2 (fr) 2010-09-16 2012-03-22 Aragon Pharmaceuticals, Inc. Modulateurs des récepteurs des oestrogènes et leurs utilisations
WO2013019906A1 (fr) 2011-08-01 2013-02-07 Genentech, Inc. Procédés de traitement du cancer à l'aide d'antagonistes se liant à l'axe pd-1 et inhibiteurs de mek
WO2013055897A1 (fr) 2011-10-14 2013-04-18 Abbvie Inc. Dérivés 8-carbamoyl-2-(2,3-di-substitué pyrid-6-yl)-1,2,3,4-tétrahydroisoquinoléine en tant qu'agents induisant une apoptose pour le traitement du cancer et de maladies immunes et auto-immunes
WO2013055895A1 (fr) 2011-10-14 2013-04-18 Abbvie Inc. Agents d'induction de l'apoptose pour traitement du cancer et de maladies immunitaires et auto-immunes
WO2013110890A1 (fr) 2012-01-24 2013-08-01 Les Laboratoires Servier Nouveaux derives d'indolizine, leur procede de preparation et les compositions pharmaceutiques qui les contiennent
US20140069822A1 (en) 2012-09-10 2014-03-13 Antec Leyden B.V. Electrochemical reduction of disulfide bonds in proteinaceous substances and electrochemical cell for carrying out such reduction
WO2014124316A2 (fr) 2013-02-08 2014-08-14 Irm Llc Sites spécifiques de modification d'anticorps pour fabriquer des immunoconjugués
WO2014130310A1 (fr) 2013-02-19 2014-08-28 Novartis Ag Dérivés de benzothiophène et compositions correspondantes en tant qu'agents de dégradation sélectifs des récepteurs des œstrogènes
WO2015011397A1 (fr) 2013-07-23 2015-01-29 Les Laboratoires Servier Nouveaux dérivés d'indolizine, leur procédé de préparation et les compositions pharmaceutiques qui les contiennent
WO2015011400A1 (fr) 2013-07-23 2015-01-29 Les Laboratoires Servier Nouveaux dérives de pyrrole, leur procédé de préparation et les compositions pharmaceutiques qui les contiennent
WO2015011396A1 (fr) 2013-07-23 2015-01-29 Les Laboratoires Servier Nouveaux derives d'indole et de pyrrole, leur procede de preparation et les compositions pharmaceutiques qui les contiennent
WO2015011164A1 (fr) 2013-07-23 2015-01-29 Les Laboratoires Servier Nouveaux composés isoindoline ou isoquinoléine, procédé pour leur préparation et compositions pharmaceutiques les contenant
WO2015011399A1 (fr) 2013-07-23 2015-01-29 Les Laboratoires Servier Nouveaux derives phosphates, leur procede de preparation et les compositions pharmaceutiques qui les contiennent
WO2015066188A1 (fr) 2013-11-01 2015-05-07 Novartis Ag Aminohétéroaryles benzamides en tant qu'inhibiteurs de kinase
WO2015097123A1 (fr) 2013-12-23 2015-07-02 Les Laboratoires Servier Nouveaux dérivés de thiénopyrimidine, procédé pour leur préparation et compositions pharmaceutiques les contenant
WO2015136017A1 (fr) 2014-03-13 2015-09-17 F. Hoffmann-La Roche Ag Méthodes et compositions pour moduler des mutants du récepteur des oestrogènes
WO2015138615A2 (fr) 2014-03-12 2015-09-17 Irm Llc Sites spécifiques utilisables pour la modification d'anticorps en vue de l'obtention d'immunoconjugués
WO2016033486A1 (fr) 2014-08-29 2016-03-03 Amgen Inc. Dérivés du tétrahydronaphtalène inhibant la protéine mcl-1
WO2016094509A1 (fr) 2014-12-09 2016-06-16 Abbvie Inc. Composés inhibiteurs de bcl xl ayant une faible perméabilité cellulaire et conjugués anticorps-médicament comprenant ceux-ci
WO2016094505A1 (fr) 2014-12-09 2016-06-16 Abbvie Inc. Conjugués anticorps médicaments avec des inhibiteurs bcl-xl à perméabilité cellulaire
WO2016094517A1 (fr) 2014-12-09 2016-06-16 Abbvie Inc. Composés inhibiteurs de bcl-xl et conjugués anticorps-médicament comprenant ceux-ci
WO2016207225A1 (fr) 2015-06-23 2016-12-29 Les Laboratoires Servier Nouveaux dérivés d'hydroxyester, leur procédé de préparation et compositions pharmaceutiques les contenant
WO2016207226A1 (fr) 2015-06-23 2016-12-29 Les Laboratoires Servier Nouveaux dérivés d'acide aminé, leur procédé de préparation et compositions pharmaceutiques les contenant
WO2016207217A1 (fr) 2015-06-23 2016-12-29 Les Laboratoires Servier Nouveaux dérivés bicycliques, leur procédé de préparation, et compositions pharmaceutiques les contenant
WO2016207216A1 (fr) 2015-06-23 2016-12-29 Les Laboratoires Servier Nouveaux dérivés d'hydroxyacide, leur procédé de préparation, et compositions pharmaceutiques les contenant
WO2017125224A1 (fr) 2016-01-19 2017-07-27 Les Laboratoires Servier Nouveaux dérivés d'ammonium, procédé de préparation de ceux-ci et compositions pharmaceutiques les contenant
WO2017147410A1 (fr) 2016-02-25 2017-08-31 Amgen Inc. Composés inhibant la protéine mcl-1
WO2017182625A1 (fr) 2016-04-22 2017-10-26 Astrazeneca Ab Inhibiteurs de mcl1 macrocycliques pour le traitement du cancer
WO2018015526A1 (fr) 2016-07-22 2018-01-25 Les Laboratoires Servier Combinaison d'un inhibiteur de bcl-2 et d'un inhibiteur de mcl-1, utilisations et compositions pharmaceutiques associées
WO2018183418A1 (fr) 2017-03-30 2018-10-04 Amgen Inc. Composés inhibant la protéine mcl-1
WO2019035927A1 (fr) 2017-08-15 2019-02-21 Abbvie Inc. Inhibiteurs de mcl-1 macrocyclique et procédés d'utilisation
WO2019035899A1 (fr) 2017-08-15 2019-02-21 Abbvie Inc. Inhibiteurs de mcl-1 macrocyclique et procédés d'utilisation
WO2019035911A1 (fr) 2017-08-15 2019-02-21 Abbvie Inc. Inhibiteurs macrocycliques de mcl-1 et méthodes d'utilisation
WO2019081559A1 (fr) 2017-10-25 2019-05-02 Les Laboratoires Servier Nouveaux derives macrocycliques, leur procede de preparation et les compositions pharmaceutiques qui les contiennent
WO2020236817A2 (fr) 2019-05-20 2020-11-26 Novartis Ag Conjugués anticorps-médicament inhibiteurs de mcl-1 et méthodes d'utilisation
WO2021018857A1 (fr) 2019-07-29 2021-02-04 Les Laboratoires Servier Dérivés de 3,6-diamino-pyridazin-3-yle, compositions pharmaceutiques les contenant et leurs utilisations en tant qu'agents pro-apoptotiques
WO2021018858A1 (fr) 2019-07-29 2021-02-04 Les Laboratoires Servier Dérivés de 6,7-dihydro-5h-pyrido[2,3-c]pyridazine et composés apparentés en tant qu'inhibiteurs de protéine bcl-xl et agents pro-apoptotiques pour le traitement du cancer
WO2022090443A1 (fr) * 2020-10-30 2022-05-05 Les Laboratoires Servier Administration et posologie pour une combinaison d'un inhibiteur de bcl-2 et d'un inhibiteur de mcl-1
WO2023161317A1 (fr) * 2022-02-24 2023-08-31 Les Laboratoires Servier Dérivés de 5-[7-(3,4-dihydro-1h-isoquinoline-2-carbonyl)-1,2,3,4 tétrahydroisoquinolin-6-y l]-1h-pyrrole-3-carboxamide, compositions pharmaceutiques les contenant et leurs utilisations en tant qu'agents pro-apoptotiques
WO2023225359A1 (fr) * 2022-05-20 2023-11-23 Novartis Ag Conjugués anticorps-médicament de composés anti-cancéreux et procédés d'utilisation

Patent Citations (68)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2779780A (en) 1955-03-01 1957-01-29 Du Pont 1, 4-diamino-2, 3-dicyano-1, 4-bis (substituted mercapto) butadienes and their preparation
US4816567A (en) 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
US6703199B1 (en) 1997-06-12 2004-03-09 Research Corporation Technologies, Inc. Artificial antibody polypeptides
WO2000035436A2 (fr) 1998-12-16 2000-06-22 Warner-Lambert Company Traitement de l'arthrite a l'aide d'inhibiteurs de la mek
WO2001051056A1 (fr) 2000-01-10 2001-07-19 Astrazeneca Ab Preparation de fulvestrant
WO2002006213A2 (fr) 2000-07-19 2002-01-24 Warner-Lambert Company Esters oxygenes d'acides 4-iodophenylamino benzhydroxamiques
WO2003076424A1 (fr) 2002-03-08 2003-09-18 Eisai Co. Ltd. Composes macrocycliques utiles comme produits pharmaceutiques
WO2003077914A1 (fr) 2002-03-13 2003-09-25 Array Biopharma, Inc Utilisation de derives de benzimidazole alkyles n3 en tant qu'inhibiteurs de mek
US7612114B2 (en) 2002-12-26 2009-11-03 Eisai R&D Management Co., Ltd. Selective estrogen receptor modulator
WO2005121142A1 (fr) 2004-06-11 2005-12-22 Japan Tobacco Inc. Dérivés de 5-amino-2,4,7-trioxo-3,4,7,8-tétrahydro-2h-pyrido’2,3-d! pyrimidine et composés apparentés pour le traitement du cancer
WO2006034488A2 (fr) 2004-09-23 2006-03-30 Genentech, Inc. Anticorps et conjugués produits avec de la cystéine
WO2007004415A1 (fr) 2005-07-01 2007-01-11 Murata Manufacturing Co., Ltd. Substrat céramique à couches multiples, procédé pour le fabriquer et feuille verte composite pour la fabrication dudit substrat
WO2008016893A1 (fr) 2006-08-02 2008-02-07 Novartis Ag Composés organiques
WO2008024725A1 (fr) 2006-08-21 2008-02-28 Genentech, Inc. Composés aza-benzofuranyle et leurs procédés d'utilisation
WO2009085983A1 (fr) 2007-12-19 2009-07-09 Genentech, Inc. 5-anilinoimidazopyridines et procédés d'utilisation
WO2010020675A1 (fr) 2008-08-22 2010-02-25 Novartis Ag Composés de pyrrolopyrimidine et leurs utilisations
US8415355B2 (en) 2008-08-22 2013-04-09 Novartis Ag Pyrrolopyrimidine compounds and their uses
US8685980B2 (en) 2008-08-22 2014-04-01 Novartis Ag Pyrrolopyrimidine compounds and their uses
WO2010080478A1 (fr) 2008-12-19 2010-07-15 Genentech, Inc. Composés et méthodes d'utilisation
WO2010080503A1 (fr) 2008-12-19 2010-07-15 Genentech, Inc. Composés hétérocycliques et leurs procédés d'utilisation
WO2011005481A1 (fr) 2009-06-22 2011-01-13 Medimmune, Llc Régions fc de synthèse pour une conjugaison spécifique à un site
WO2011076786A1 (fr) 2009-12-22 2011-06-30 Novartis Ag Isoquinolinones et quinazolinones substituées
WO2011156518A2 (fr) 2010-06-10 2011-12-15 Aragon Pharmaceuticals, Inc. Modulateur du récepteur oestrogénique et utilisation de ces derniers
WO2011159769A2 (fr) 2010-06-17 2011-12-22 Aragon Pharmaceuticals, Inc. Modulateurs de récepteur d'œstrogène d'indane et utilisations de ceux-ci
WO2012037410A2 (fr) 2010-09-16 2012-03-22 Aragon Pharmaceuticals, Inc. Modulateurs des récepteurs des oestrogènes et leurs utilisations
US20120071535A1 (en) 2010-09-16 2012-03-22 Aragon Pharmaceuticals, Inc. Estrogen receptor modulators and uses thereof
WO2012037411A2 (fr) 2010-09-16 2012-03-22 Aragon Pharmaceuticals, Inc. Modulateurs des récepteurs des oestrogènes et leurs utilisations
WO2013019906A1 (fr) 2011-08-01 2013-02-07 Genentech, Inc. Procédés de traitement du cancer à l'aide d'antagonistes se liant à l'axe pd-1 et inhibiteurs de mek
WO2013055897A1 (fr) 2011-10-14 2013-04-18 Abbvie Inc. Dérivés 8-carbamoyl-2-(2,3-di-substitué pyrid-6-yl)-1,2,3,4-tétrahydroisoquinoléine en tant qu'agents induisant une apoptose pour le traitement du cancer et de maladies immunes et auto-immunes
WO2013055895A1 (fr) 2011-10-14 2013-04-18 Abbvie Inc. Agents d'induction de l'apoptose pour traitement du cancer et de maladies immunitaires et auto-immunes
WO2013110890A1 (fr) 2012-01-24 2013-08-01 Les Laboratoires Servier Nouveaux derives d'indolizine, leur procede de preparation et les compositions pharmaceutiques qui les contiennent
US20140069822A1 (en) 2012-09-10 2014-03-13 Antec Leyden B.V. Electrochemical reduction of disulfide bonds in proteinaceous substances and electrochemical cell for carrying out such reduction
WO2014124316A2 (fr) 2013-02-08 2014-08-14 Irm Llc Sites spécifiques de modification d'anticorps pour fabriquer des immunoconjugués
WO2014130310A1 (fr) 2013-02-19 2014-08-28 Novartis Ag Dérivés de benzothiophène et compositions correspondantes en tant qu'agents de dégradation sélectifs des récepteurs des œstrogènes
WO2015011397A1 (fr) 2013-07-23 2015-01-29 Les Laboratoires Servier Nouveaux dérivés d'indolizine, leur procédé de préparation et les compositions pharmaceutiques qui les contiennent
WO2015011400A1 (fr) 2013-07-23 2015-01-29 Les Laboratoires Servier Nouveaux dérives de pyrrole, leur procédé de préparation et les compositions pharmaceutiques qui les contiennent
WO2015011396A1 (fr) 2013-07-23 2015-01-29 Les Laboratoires Servier Nouveaux derives d'indole et de pyrrole, leur procede de preparation et les compositions pharmaceutiques qui les contiennent
WO2015011164A1 (fr) 2013-07-23 2015-01-29 Les Laboratoires Servier Nouveaux composés isoindoline ou isoquinoléine, procédé pour leur préparation et compositions pharmaceutiques les contenant
WO2015011399A1 (fr) 2013-07-23 2015-01-29 Les Laboratoires Servier Nouveaux derives phosphates, leur procede de preparation et les compositions pharmaceutiques qui les contiennent
WO2015066188A1 (fr) 2013-11-01 2015-05-07 Novartis Ag Aminohétéroaryles benzamides en tant qu'inhibiteurs de kinase
WO2015097123A1 (fr) 2013-12-23 2015-07-02 Les Laboratoires Servier Nouveaux dérivés de thiénopyrimidine, procédé pour leur préparation et compositions pharmaceutiques les contenant
WO2015138615A2 (fr) 2014-03-12 2015-09-17 Irm Llc Sites spécifiques utilisables pour la modification d'anticorps en vue de l'obtention d'immunoconjugués
WO2015136017A1 (fr) 2014-03-13 2015-09-17 F. Hoffmann-La Roche Ag Méthodes et compositions pour moduler des mutants du récepteur des oestrogènes
WO2016033486A1 (fr) 2014-08-29 2016-03-03 Amgen Inc. Dérivés du tétrahydronaphtalène inhibant la protéine mcl-1
WO2016094509A1 (fr) 2014-12-09 2016-06-16 Abbvie Inc. Composés inhibiteurs de bcl xl ayant une faible perméabilité cellulaire et conjugués anticorps-médicament comprenant ceux-ci
WO2016094505A1 (fr) 2014-12-09 2016-06-16 Abbvie Inc. Conjugués anticorps médicaments avec des inhibiteurs bcl-xl à perméabilité cellulaire
WO2016094517A1 (fr) 2014-12-09 2016-06-16 Abbvie Inc. Composés inhibiteurs de bcl-xl et conjugués anticorps-médicament comprenant ceux-ci
WO2016207225A1 (fr) 2015-06-23 2016-12-29 Les Laboratoires Servier Nouveaux dérivés d'hydroxyester, leur procédé de préparation et compositions pharmaceutiques les contenant
WO2016207226A1 (fr) 2015-06-23 2016-12-29 Les Laboratoires Servier Nouveaux dérivés d'acide aminé, leur procédé de préparation et compositions pharmaceutiques les contenant
WO2016207217A1 (fr) 2015-06-23 2016-12-29 Les Laboratoires Servier Nouveaux dérivés bicycliques, leur procédé de préparation, et compositions pharmaceutiques les contenant
WO2016207216A1 (fr) 2015-06-23 2016-12-29 Les Laboratoires Servier Nouveaux dérivés d'hydroxyacide, leur procédé de préparation, et compositions pharmaceutiques les contenant
WO2017125224A1 (fr) 2016-01-19 2017-07-27 Les Laboratoires Servier Nouveaux dérivés d'ammonium, procédé de préparation de ceux-ci et compositions pharmaceutiques les contenant
WO2017147410A1 (fr) 2016-02-25 2017-08-31 Amgen Inc. Composés inhibant la protéine mcl-1
WO2017182625A1 (fr) 2016-04-22 2017-10-26 Astrazeneca Ab Inhibiteurs de mcl1 macrocycliques pour le traitement du cancer
WO2018015526A1 (fr) 2016-07-22 2018-01-25 Les Laboratoires Servier Combinaison d'un inhibiteur de bcl-2 et d'un inhibiteur de mcl-1, utilisations et compositions pharmaceutiques associées
WO2018183418A1 (fr) 2017-03-30 2018-10-04 Amgen Inc. Composés inhibant la protéine mcl-1
WO2019035927A1 (fr) 2017-08-15 2019-02-21 Abbvie Inc. Inhibiteurs de mcl-1 macrocyclique et procédés d'utilisation
WO2019035899A1 (fr) 2017-08-15 2019-02-21 Abbvie Inc. Inhibiteurs de mcl-1 macrocyclique et procédés d'utilisation
US20190055264A1 (en) 2017-08-15 2019-02-21 Abbvie Inc. Macrocyclic MCL-1 inhibitors and methods of use
WO2019035914A1 (fr) 2017-08-15 2019-02-21 Abbvie Inc. Inhibiteurs de mcl-1 macrocyclique et procédés d'utilisation
WO2019035911A1 (fr) 2017-08-15 2019-02-21 Abbvie Inc. Inhibiteurs macrocycliques de mcl-1 et méthodes d'utilisation
WO2019081559A1 (fr) 2017-10-25 2019-05-02 Les Laboratoires Servier Nouveaux derives macrocycliques, leur procede de preparation et les compositions pharmaceutiques qui les contiennent
WO2020236817A2 (fr) 2019-05-20 2020-11-26 Novartis Ag Conjugués anticorps-médicament inhibiteurs de mcl-1 et méthodes d'utilisation
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Non-Patent Citations (84)

* Cited by examiner, † Cited by third party
Title
"Remington's Pharmaceutical Sciences", 1990, ED. MACK PRINTING COMPANY, pages: 1289 - 1329
"Roche Molecular Biochemicals", BIOCHEMICA, vol. 2, 1999, pages 34 - 7
"UniProt", Database accession no. Q07817-1
AB ET AL., MOL CANCER THER., vol. 14, 2015, pages 1605 - 13
ACKLER ET AL., CANCER CHEMOTHER. PHARMACOL., vol. 66, no. 5, pages 869 - 80
AL-LAZIKANI ET AL., J MOL BIOL., vol. 273, no. 4, 1997, pages 927 - 48
AMUNDSON ET AL., CANCER RES., vol. 60, no. 21, 1 November 2000 (2000-11-01), pages 6101 - 10
BAH ET AL., CELL DEATH AND DISEASE,, vol. 5, 2014, pages 1291
BARDWELL ET AL., J CLIN INVEST., vol. 99, no. 3, 1 February 1997 (1997-02-01), pages 439 - 46
BENNETT ET AL., OPEN BIOL.,, vol. 6, 2016, pages 160134
BERGE ET AL.: "Pharmaceutical Salts", J. PHARMACEUTICAL SCIENCES, vol. 66, no. 1, 1977, XP002675560, DOI: 10.1002/jps.2600660104
BEROUKHIM ET AL., NATURE, vol. 463, no. 7283, 18 February 2010 (2010-02-18), pages 899 - 905
BOER K. ET AL., THERAPEUTIC ADVANCES IN MEDICAL ONCOLOGY, vol. 9, no. 7, 2017, pages 465 - 479
BRICOGNE ET AL.: "BUSTER", 2017, GLOBAL PHASING LTD.
CAENEPEEL ET AL., CANCER DISCOV., vol. 8, no. 12, December 2018 (2018-12-01), pages 1582 - 1597
CASARA ET AL., ONCOTARGET, vol. 9, no. 28, 13 April 2018 (2018-04-13), pages 20075 - 20088
CHEN ET AL., MOL. CANCER THER., vol. 10, no. 12, December 2011 (2011-12-01), pages 2340 - 9
CIPPA ET AL., TRANSPL INT., vol. 24, no. 7, July 2011 (2011-07-01), pages 722 - 32
CLACKSON, NATURE, vol. 352, 1991, pages 624 - 8
CZABOTAR ET AL., NAT. REV. MOL. CELL BIOL., vol. 15, no. 1, January 2014 (2014-01-01), pages 49 - 63
DAMLE, N.K., NAT BIOTECHNOL., vol. 26, no. 1204531-26-9, 2008, pages 884 - 885
DROIN ET AL., BIOCHIM BIOPHYS ACTA, 1 March 2004 (2004-03-01)
DUCRYSTUMP, BIOCONJUGATE CHEM., vol. 21, 2010, pages 5 - 13
DUKECOHEN ET AL.: "Current Protocols in Immunology", 1992, pages: 1 - 16
EMSLEY AND COWTAN: "Acta Crystallogr. D. Biol. Crystallogr.", CRYSTALLOGR., vol. 60, no. 12, 2004, pages 2126 - 32
FINN ET AL., BREAST CANCER RESEARCH, vol. 11, no. 571190-30-2, 2009, pages R77
GARNER F ET AL., ANTICANCER DRUGS, vol. 26, no. 722533-56-4, 2015, pages 948 - 56
GERSHONI ET AL., BIODRUGS, vol. 21, 2007, pages 145 - 56
GROSS ET AL., GENES DEV., vol. 13, no. 15, 1 August 1999 (1999-08-01), pages 1899 - 911
GRUNDY ET AL., ONCOTARGET, vol. 9, no. 102, 28 December 2018 (2018-12-28), pages 37777 - 37789
HAGER-BRAUNTOMER, EXPERT REV PROTEOMICS, vol. 2, 2005, pages 745 - 56
HANAHANWEINBERG, CELL, vol. 100, no. 1, 7 January 2000 (2000-01-07), pages 57 - 70
HANAHANWEINBERG, CELL, vol. 144, no. 5, 4 March 2011 (2011-03-04), pages 646 - 74
HAYNES ET AL.: "ommentary: Occurrence of Pharmaceutically Acceptable Anions and Cations in the Cambridge Structural Database", J. PHARMACEUTICAL SCIENCES, vol. 94, no. 10, 2005, XP002593272
HOLLIGERHUDSON, NAT BIOTECHNOL., vol. 23, no. 9, 2005, pages 1126 - 36
J. ORG. CHEM., vol. 77, 2012, pages 4226 - 4234
JUNUTULA JR ET AL., NAT BIOTECHNOL, vol. 26, 2008, pages 925 - 932
KLEIN ET AL., NATURE MED., vol. 3, 1997, pages 402 - 8
KNAPPIK, J MOL BIOL., vol. 296, no. 1, 2000, pages 57 - 86
KOHLER ET AL., NATURE, vol. 256, 1975, pages 495
KOTSCHY ET AL., NATURE, vol. 538, 2016, pages 477 - 482
LEFRANC ET AL., DEV COMP IMMUNOL., vol. 27, no. 1, 2003, pages 55 - 77
LEFRANC, NUCLEIC ACIDS RES., vol. 29, no. 1, 2001, pages 207 - 9
LEVERSON ET AL., CELL DEATH DIS., vol. 6, 15 January 2015 (2015-01-15), pages e1590
LEVERSON ET AL., SCI TRANSL MED., vol. 7, no. 279, 18 March 2015 (2015-03-18), pages 279 - 40
LEVERSON ET AL., SCI. TRANSL. MED., vol. 7, no. 279, 18 March 2015 (2015-03-18), pages 279 - 40
LEVERSON ET AL., SCIENCE TRANSLATION MEDICINE, vol. 7, no. 279, 18 March 2015 (2015-03-18), pages 279 - 40
LYON ET AL., METHODS ENZYMOL., vol. 502, 2012, pages 123 - 38
LYONS ET AL., PROTEIN ENG., vol. 3, 1990, pages 703 - 708
MARAGNO ET AL., AACR, 2019
MARKS ET AL., J MOL BIOL., vol. 222, 1991, pages 581 - 97
MARSDENSTRASSER, ANNU REV IMMUNOL., vol. 21, 2003, pages 71 - 105
MCCOY ET AL., J. APPL. CRYSTALLOGR., vol. 40, no. 4, 2007, pages 658 - 674
MCDONELL, JOURNAL OF MEDICINAL CHEMISTRY, vol. 58, no. 1365888-06-7, 2015, pages 4883 - 4887
MERINO ET AL., APOPTOSIS, vol. 14, no. 4, April 2009 (2009-04-01), pages 570 - 83
MERINO ET AL., SCIENCE TRANSLATIONAL MEDICINE, vol. 9, no. 401, 2 August 2017 (2017-08-02), pages eaam7049
MEYERSMILLER, CABIOS, vol. 4, 1989, pages 11 - 17
MOUJALLED ET AL., BLOOD ADV., vol. 4, no. 12, 23 June 2020 (2020-06-23), pages 2762 - 2767
MOUJALLED ET AL., LEUKEMIA, vol. 33, no. 4, April 2019 (2019-04-01), pages 905 - 917
NEEDLEMANWUNSCH, J MOL BIOL., vol. 48, 1970, pages 444 - 53
NGUYEN ET AL., CLIN CANCER RES, vol. 17, no. 6, 15 March 2011 (2011-03-15), pages 1394 - 1404
PAGE ET AL., INTL J ONCOLOGY, vol. 3, 1993, pages 473 - 6
PARK ET AL., J. MED. CHEM., vol. 51, no. 21, 13 November 2008 (2008-11-13), pages 6902 - 15
POPE ET AL., NAT REV IMMUNOL., vol. 2, no. 7, July 2002 (2002-07-01), pages 527 - 35
ROBERTS ET AL., J. CLIN. ONCOL., vol. 30, no. 5, 10 February 2012 (2012-02-10), pages 488 - 96
SARAH KEHR ET AL., CANCER LETT., vol. 482, 10 July 2020 (2020-07-10), pages 19 - 32
SHOEMAKER ET AL., CLIN. CANCER RES., vol. 14, no. 11, 1 June 2008 (2008-06-01), pages 3268 - 77
SINGH, S.K, PHARM RES., vol. 32, no. 11, 2015, pages 3541 - 71
SODERQUIST ET AL., NAT COMMUN., vol. 9, no. 1, 29 August 2018 (2018-08-29), pages 3513
SOUERS ET AL., NAT MED., vol. 19, no. 2, February 2013 (2013-02-01), pages 202 - 8
STEFANO ET AL., METHODS MOL BIOL., vol. 1045, 2013, pages 145 - 71
TAO ET AL., ACS MED CHEM LETT., vol. 5, no. 10, 26 August 2014 (2014-08-26), pages 1088 - 93
TAO ET AL., ACS MED. CHEM. LETT., vol. 5, no. 10, 26 August 2014 (2014-08-26), pages 1088 - 93
TAO ET AL., ACS MEDICINAL CHEMISTRY LETTERS, vol. 5, no. 10, 2014, pages 1088 - 109
TOPHAM ET AL., CANCER CELL,, vol. 28, 2015, pages 129 - 140
TORRES-GUZMAN R ET AL., ONCOTARGET, no. 1231929-97-7, 2017
TRON ET AL., NAT. COMMUN., vol. 9, no. 1, 17 December 2018 (2018-12-17), pages 5341
TZARTOS, METHODS IN MOLECULAR BIOLOG, vol. 66, 1998, pages 55 - 66
VAUXFLAVELL, CURR OPIN IMMUNOL., vol. 12, no. 6, December 2000 (2000-12-01), pages 719 - 24
WANG ET AL., ACS MEDICINAL CHEMISTRY LETTERS, vol. 11, no. 10, 2020, pages 1829 - 1826
WEEDEN ET AL., ONCOGENE, vol. 37, no. 32, August 2018 (2018-08-01), pages 4475 - 4488
WILSON ET AL., LANCET ONCOL, vol. 11, no. 12, December 2010 (2010-12-01), pages 1149 - 59
WONG ET AL., MOL CANCER THER, vol. 11, no. 4, April 2012 (2012-04-01), pages 1026 - 1035
YOULESTRASSER, NAT. REV. MOL. CELL BIOL., vol. 9, no. 1, January 2008 (2008-01-01), pages 47 - 59

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