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US-12617866-B2 - Humanized antibodies to mucin-16 and methods of use thereof

US12617866B2US 12617866 B2US12617866 B2US 12617866B2US-12617866-B2

Abstract

Provided herein are compositions, methods, and uses involving anti-Mucin-16 (MUC16) agents that immunospecifically bind an epitope of Mucin-16 (MUC16). Also provided herein are uses and methods for managing, treating, or preventing disorders, such as cancer and diseases associated with positive MUC16 expression.

Inventors

  • David Spriggs
  • Dharmarao Thapi
  • Su Yan
  • Cheng Liu

Assignees

  • MEMORIAL SLOAN KETTERING CANCER CENTER
  • Eureka Therapeutics, Inc.

Dates

Publication Date
20260505
Application Date
20200507

Claims (12)

  1. 1 . An anti-mucin 16 (MUC16) construct comprising an antibody moiety that immunospecifically recognizes a mucin 16 (MUC16) polypeptide, wherein the antibody moiety comprises: (a) (i) a variable heavy (VH) chain comprising SEQ ID NO: 4 or 5; and (ii) a variable light (VL) chain comprising SEQ ID NO: 2 or 3; or (b) (i) a variable heavy (VH) chain comprising SEQ ID NO: 22 or 23; and (ii) a variable light (VL) chain comprising SEQ ID NO: 20 or 21, wherein the VH chain and VL chain are humanized.
  2. 2 . The anti-MUC16 construct of claim 1 , wherein the antibody moiety (a) immunospecifically binds to the ectodomain of MUC16, or to a MUC16 c114 polypeptide comprising the amino acid sequence of SEQ ID NO: 44 or 180; (b) is a full-length antibody, a monoclonal antibody, a Fab, a Fab′, a F(ab′)2, an a Fv, or a single chain Fv (scFv), wherein the scFv comprises any one of SEQ ID NOs: 53-68; (c) comprises human-derived heavy and light chain constant regions, wherein the heavy chain constant region has an isotype selected from the group consisting of gamma 1, gamma 2, gamma 3, and gamma 4, or wherein the light chain constant region has an isotype selected from the group consisting of kappa and lambda; or (d) is an immunoglobulin comprising two identical heavy chains and two identical light chains, wherein the immunoglobulin is an IgG.
  3. 3 . The anti-MUC16 construct of claim 1 , wherein the anti-MUC16 construct inhibits in vitro invasion of a MUC16-expressing tumor cell in a gel invasion assay, wherein the MUC16-expressing tumor cell is an ovarian tumor cell; or is monospecific, multispecific, or bispecific, wherein the multispecific or bispecific anti-MUC16 construct comprises an anti-CD3 antibody moiety; or is (i) a tandem scFv, wherein the tandem scFv comprises two scFvs linked by a peptide linker; (ii) a diabody (Db); (iii) a single chain diabody (scDb); (iv) a dual-affinity retargeting (DART) antibody, (v) a F(ab′)2; (vi) a dual variable domain (DVD) antibody; (vii) a knob-into-hole (KiH) antibody; (viii) a dock and lock (DNL) antibody; (ix) a chemically cross-linked antibody; (x) a heteromultimeric antibody; or (xi) a heteroconjugate antibody; or is a chimeric antigen receptor (CAR) comprising at least one of: (i) a co-stimulatory domain, (ii) a CD3 zeta (ζ) chain cytoplasmic signaling domain, (iii) an scFv of any a one of SEQ ID NOS: 53-68, or (iv) any one of SEQ ID NOS: 80-87 and 97-103.
  4. 4 . The anti-MUC16 construct of claim 3 , wherein the multispecific or bispecific anti-MUC16 construct comprises a first antibody moiety that immunospecifically recognizes MUC16, and a second antibody moiety that immunospecifically a recognizes a second antigen, wherein the second antigen is a CD3 polypeptide selected from the group consisting of CD3γ, CD3δ, CD3ε, and CD3ζ or wherein the anti-MUC16 construct comprises any one of SEQ ID NOS: 42, 69-75, and 88-95.
  5. 5 . The anti-MUC16 construct of claim 1 , further conjugated to a peptide agent, a detection agent, an imaging agent, a therapeutic agent, a cytotoxic agent, an alpha emitter, an Auger-emitter, a beta-emitter, a gamma-emitter, a positron-emitter, or an x-ray a emitter.
  6. 6 . A polypeptide comprising an amino acid sequence of the anti-MUC16 construct of claim 1 .
  7. 7 . A polynucleotide or vector comprising a nucleic acid sequence encoding one or more polypeptides of claim 6 , wherein the nucleic acid sequence is operably linked to a promoter.
  8. 8 . A cell comprising the polynucleotide or vector of claim 7 , wherein the cell is a mammalian cell, an immune cell, a lymphocyte, a T cell or a B cell.
  9. 9 . A pharmaceutical composition comprising: a a therapeutically effective amount of the anti-MUC16 construct of claim 1 ; and a pharmaceutically acceptable carrier.
  10. 10 . A method of treating a MUC16-associated disease or disorder in a patient in need thereof, comprising administering to said patient a a therapeutically effective amount of the anti-MUC16 construct of claim 1 , wherein said MUC16-associated disease or disorder is a metastatic cancer or a cancer of the ovary, lung, pancreas, breast, uterine, fallopian tube, or primary peritoneum.
  11. 11 . A method of detecting MUC16 in a sample, a comprising: (a) contacting the sample with the anti-MUC16 construct of claim 1 ; and (b) detecting direct or indirect binding between the anti-MUC16 construct and a MUC16 polypeptide in the sample, wherein the anti-MUC16 construct is conjugated to a detectable label selected from among a chromogenic label, an enzymatic label, a radioisotopic label, an isotopic label, a fluorescent label, a toxic label, a chemiluminescent label, and a nuclear magnetic resonance contrast agent.
  12. 12 . A method for detecting cancer in a subject in vivo comprising (a) administering to the subject an effective amount of the anti-MUC16 construct a of claim 1 , wherein the anti-MUC16 construct is configured to localize to a cancer cell expressing MUC16 and is labeled with a radioisotope; and (b) detecting the presence of a tumor in the subject by detecting radioactive levels emitted by the anti-MUC16 construct that are higher than a reference value, wherein the radioisotope is 89Zr-desferrioxamine B (DFO) or wherein the radioactive levels emitted by the anti-MUC16 construct are detected using positron emission tomography or single photon emission computed tomography.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a U.S. National Stage Application under 35 U.S.C. § 371 of International Patent Application No. PCT/US2020/031886, filed on May 7, 2020, which claims the benefit of and priority to U.S. Provisional Patent Application No. 62/845,065, filed May 8, 2019, the entire contents of each of which are incorporated herein by reference. STATEMENT OF GOVERNMENT SUPPORT This invention was made with government support under P01 CA190174-01, P01 CA190174-02, and P01 CA190174-03, awarded by the National Institutes of Health. The government has certain rights in the invention. SEQUENCE LISTING The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Jun. 12, 2020, is named 115872-0832_SL.txt and is 545,591 bytes in size. BACKGROUND Mucins are important biomolecules for cellular homeostasis and protection of epithelial surfaces. Changes in expression of mucins in cancers, such as ovarian cancer, are useful as a biomarker for diagnosis, prognosis and treatment (Singh A P, et al., Lancet Oncol 2008; 9(11): 1076-85). MUC16 is a mucin that is over expressed on most ovarian carcinoma cells and is an established surrogate serum marker (CA-125) for the detection and progression of ovarian cancers (Badgwell D, et al., Dis Markers 23(5-6):397410 (2007); Bast RC, Jr, et al., Int J Gynecol Cancer 15 Suppl 3:274-81 (2005); Fritsche H A, et al., Clin Chem 44(7): 1379-80 (1998); and Krivak T C et al., Gynecol Oncol 115(1):81-5 (2009)). MUC16 is a highly glycosylated mucin composed of a large extracellular domain (CA-125), which is cleaved and released, and a retained domain (MUC-CD) (FIG. 1). MUC-CD comprises a non-repeating extracellular domain (MUC16 ectodomain) proximal to a cleavage site, a transmembrane domain, and a cytoplasmic tail with potential phosphorylation sites. Distal to the cleavage site, the released extracellular domain (CA-125) contains 16-20 tandem repeats of 156 amino acids, each with many potential glycosylation sites (O'Brien T J, et al., Tumor Biol 22(6):348-66 (2001)). Since the MUC16 antigen is otherwise expressed only at low levels in normal tissues of the uterus, endometrium, fallopian tubes, ovaries, and serosa of the abdominal and thoracic cavities, MUC16 is a potentially attractive target for immune-based therapies, including the targeting and treatment of cancer. A significant portion of the extracellular domain of MUC16 is cleaved and secreted (i.e., CA-125), which limits the utility of this portion of MUC16 to be used as a target antigen on ovarian carcinomas. Many reported MUC16 monoclonal antibodies bind to epitopes present on the large secreted CA-125 fraction of the glycoprotein, and not to the retained MUC16 ectodomain (Bellone S Am J Obstet Gynecol 200(1):75 el-10 (2009), Berek J S. Expert Opin Biol Ther. 4(7): 1159-65 (2004); O'Brien T J, et al., Int J Biol Markers 13(4): 188-95 (1998)). Thus, the generation of new antibodies to the region of MUC16 that is not shed are needed for diagnostic and therapeutic purposes. SUMMARY OF THE PRESENT TECHNOLOGY Provided herein are compositions, methods, and uses of anti-Mucin 16 (MUC16) constructs that comprise antibody moieties that immunospecifically bind to Mucin 16 (MUC16), and modulate expression and/or activity of MUC16 for managing or treating MUC16-mediated disorders, such as cancer. Provided herein, in certain embodiments, are anti-mucin 16 (MUC16) constructs comprising an antibody moiety that immunospecifically recognizes a mucin 16 (MUC16) polypeptide, wherein the antibody moiety comprises a humanized heavy chain variable domain and a humanized light chain variable domain of a 4H11 or 18C6 murine monoclonal antibody. In some embodiments, the antibody moiety comprises (a) (i) a variable heavy (VH) chain comprising a heavy chain complementarity determining region 1 (HC-CDR1), HC-CDR2, and HC-CDR3 of SEQ ID NOS: 17, 18, and 19, respectively, and a heavy chain framework region 1 (HC-FW1), HC-FW2, and HC-FW3 of SEQ ID NOS: 136, 137, and 138, respectively, wherein one or more amino acids selected from amino acid positions 1, 3, 5, 11 and 19 of SEQ ID NO: 136, amino acid positions 5, 7, 8, and 9 of SEQ ID NO: 137, and amino acid positions 12, 14, 18, 22, and 23 of SEQ ID NO: 138 is humanized relative to a mouse HC-FW1, HC-FW2, and HC-FW3 of SEQ ID NOS: 124, 125, and 126, respectively; (ii) a variable light (VL) chain comprising a light chain complementarity determining region 1 (LC-CDR1), LC-CDR2, and LC-CDR3 of SEQ ID NOS: 14, 15, and 16, respectively, and a light chain framework region 1 (LC-FW1), LC-FW2, LC-FW3, and LC-FW4 of SEQ ID NOS: 120, 121, 122, and 123, respectively, wherein one or more amino acids selected from positions 3, 9, 15, 18, and 22 of SEQ ID NO: 120, amino acid positions 7 and 27 of SEQ ID NO: 122, and amino acid positions 3 and 9 of SEQ ID NO