Search

CN-121986119-A - Humanized MUC1 antibodies and antibody drug conjugates

CN121986119ACN 121986119 ACN121986119 ACN 121986119ACN-121986119-A

Abstract

The present invention relates to humanized MUC1 antibodies, antibody Drug Conjugates (ADCs) comprising such antibodies, and their use for treating conditions such as cancer, particularly in the case of MUC1 overexpression.

Inventors

  • N. K. Damel
  • J. Shakti
  • N. KUMAR
  • V. B. Mehra
  • A. P. Beira
  • S. GUPTA
  • V. P. Kumar
  • A. TUCKER
  • R. Churuwatier
  • A. Darth
  • B. Samantha

Assignees

  • 太阳医药高级研究有限公司

Dates

Publication Date
20260505
Application Date
20240731
Priority Date
20230731

Claims (20)

  1. 1. A humanized monoclonal antibody that binds to a MUC1 SEA domain, wherein the antibody comprises: (a) A heavy chain variable region comprising the amino acid sequence of any one of SEQ ID NOS.7-11, 45 and 46, and (B) A light chain variable region comprising the amino acid sequence of any one of SEQ ID nos. 12-15.
  2. 2. The humanized antibody of claim 1, wherein the antibody comprises: (a) A heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 8 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 12; (b) A heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 8 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 13; (c) A heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 8 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 14; (d) A heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 9 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 13; (e) A heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 9 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 14; (f) A heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 10 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 13; (g) A heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 10 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 14; (h) A heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 11 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 13; (i) A heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 11 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 14, or (J) A heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 11 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 15.
  3. 3. The humanized antibody according to claim 1 or 2, wherein the constant domain of the heavy chain comprises the amino acid sequence of SEQ ID No. 39 and the constant domain of the light chain comprises the amino acid sequence of SEQ ID No. 40.
  4. 4. A humanized monoclonal antibody that binds to the MUC1 SEA domain, wherein the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID No. 11 and a light chain variable region comprising the amino acid sequence of SEQ ID No. 15.
  5. 5. A humanized monoclonal antibody that binds to the MUC1 SEA domain, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID No. 37 and a light chain comprising the amino acid sequence of SEQ ID No. 38.
  6. 6. The humanized antibody of claim 5, wherein part of Conjugated to the light chain of the humanized antibody through its maleimide terminus.
  7. 7. The humanized monoclonal antibody of claim 5 or 6, wherein the heavy chain has the amino acid sequence of SEQ ID No. 37 and the light chain has the amino acid sequence of SEQ ID No. 38.
  8. 8. The humanized monoclonal antibody of any of the preceding claims, wherein the antigen binding fragment thereof is Fv, single chain Fv (scFv), single chain Fv-Fc (scFv-Fc), fab ', fab, F (ab') 2 or F (ab) 2 .
  9. 9. A humanized antibody that binds to an epitope in the MUC1 SEA domain, preferably having a binding affinity K D of less than 100 pM.
  10. 10. The humanized antibody of claim 9, wherein the humanized antibody comprises means for binding to an epitope in the SEA domain of MUC1 (SEQ ID NO: 41) formed by arginine at position 1108, glutamic acid at position 1109, asparagine at position 1113 and glutamic acid at position 1118 of MUC 1.
  11. 11. The humanized antibody according to claim 9 or 10, wherein the humanized antibody binds to the same epitope as a chimeric antibody comprising a heavy chain variable region and a light chain variable region comprising the amino acid sequences of SEQ ID NOs 1 and 2, respectively.
  12. 12. The humanized antibody of claim 9 or 10, wherein the antibody is a humanized version of any of the chimeric antibodies DMB4F4 (4F 4), DMB7F3 (7F 3) or DMB10F10 (10F 10).
  13. 13. A humanized antibody that binds to an epitope in the SEA domain of MUC1 (SEQ ID NO: 41) that is formed from arginine at position 1108, glutamic acid at position 1109, asparagine at position 1113, and glutamic acid at position 1118 of MUC 1.
  14. 14. A humanized antibody that binds to the same epitope as a chimeric antibody comprising a heavy chain variable region and a light chain variable region comprising the amino acid sequences of SEQ ID NOs 1 and 2, respectively.
  15. 15. An isolated nucleic acid molecule comprising a nucleotide sequence encoding the antibody of any one of the preceding claims.
  16. 16. An expression vector comprising the isolated nucleic acid molecule of claim 15.
  17. 17. A host cell transfected with the expression vector of claim 16.
  18. 18. An immunoconjugate comprising the antibody of any one of claims 1 to 14, and an additional cytotoxic or therapeutic agent.
  19. 19. The immunoconjugate according to claim 18, wherein the cytotoxic agent is selected from the group consisting of an alkylating drug, an anthracycline, a pyrimidine derivative, a vinca alkaloid, a photodynamic drug, a platinum-containing compound, a taxane, a topoisomerase inhibitor, a ribosome inactivating agent, a DNA damage inducing agent, a tubulin inhibitor, an antimitotic agent, a radioisotope, a cytotoxic antibody, and a bacterial toxin.
  20. 20. The immunoconjugate according to claim 18, wherein the cytotoxic agent is a pseudomonas (pseudomonas) exotoxin.

Description

Humanized MUC1 antibodies and antibody drug conjugates The present application claims the benefit of indian patent application number 202321051338 filed on 7/31 2023, which is hereby incorporated by reference. The computer readable form of the sequence listing is submitted with the present application by way of electronic submission and is hereby incorporated by reference in its entirety. The ST26 sequence Listing is contained in the file created on month 7 and 31 of 2024 under the file name "159818-02601_SL.xml". Technical Field The present invention relates to humanized MUC1 antibodies, antibody Drug Conjugates (ADCs) comprising such antibodies, and their use for treating conditions such as cancer, particularly in the case of MUC1 overexpression. Background MUC1 glycoprotein is overexpressed by a variety of high-incidence, high-mortality human epithelial malignancies (including breast, prostate, pancreatic, ovarian, lung and colon cancers), as well as by malignant plasma cells of multiple myeloma and myeloid cells of acute myelogenous leukemia. MUC1 has been studied as a target for targeted cancer therapy and as a marker of disease progression, since it is preferentially highly expressed by malignant cells, and since it is expressed on the cell surface and thus is a naked molecule. RIVALLAND et al, expert opinion on biological therapy (Expert Opin Biol Ther), 15 (2015) 1773-1787; taylor-Papadimitriou et al, society of biochemistry (Biochem Soc Trans), 46 (2018) 659-668. Structurally, the MUC1 molecule is a transmembrane glycoprotein (known as MUC-TM). MUC-TM is a heterodimer consisting of an extracellular domain comprising 20 to 125 or more repeats of a 20 amino acid long sequence (known as variable number of tandem repeats, VNTR), a transmembrane domain, and a short cytoplasmic tail that mediates intracellular signaling. The MUC1 molecule is self-proteolytically cleaved within the SEA (sperm protein, enterokinase and agrin) module, which is a highly conserved domain of 110 amino acids. This results in the binding of large extracellular alpha subunits containing tandem repeat arrays to transmembrane beta subunits containing the transmembrane and cytoplasmic domains of the molecule with strong non-covalent interactions. The binding of the alpha chain to the beta chain is intermittent in that the alpha chain binds to the beta chain in an intermittent manner. The alpha chain with its VNTR only intermittently maintains cell binding when the beta chain remains on the cell surface at all times. Many anti-MUC 1 antibodies have been reported in the literature, most of which are directed against highly immunogenic VNTR for the alpha chain. Although anti-VNTR antibodies can successfully bind muc1+ cells in vitro, the shedding of the muc1α chain containing VNTR into the peripheral circulation in vivo severely compromises the ability of anti-VNTR antibodies to clinically affect MUC1 expressing tumors. Not only does the shedding of alpha chains from the tumor cell surface greatly reduce the number of MUC1 targets of anti-alpha chain antibodies, but in addition, free circulating MUC1 alpha chains in the peripheral circulation in vivo can bind and neutralize anti-VNTR antibodies or anti-glycosylated VNTR antibodies, thereby limiting their ability to even reach MUC1 expressing tumors. Antibodies recognizing cancer-specific truncated O glycoforms of VNTR (such as antibodies PankoMab-Gex, 5E5, SM3 and VU-2-G7) have been proposed as a possible way to overcome the potential toxicity of targeting MUC1 expressed by normal tissues. However, limitations on targeting the alpha chain VNTR, i.e., its shedding from the cell surface and its ability to bind therapeutically administered anti-MUC 1 antibodies, remain. Burchell et al, (J MAMMARY GLAND Biol Neoplasia) journal of breast biology and tumors, 6 (2001) 355-364; fiedler et al, (Eur J Cancer), 63 (2016) 55-63; ryuko et al, (Tumour Biol) journal of tumor biology, 21 (2000) 197-210; tarp et al, (Glycobiology), 17 (2007) 197-209; zhou et al, (Molecules) molecule, 23 (6) (2018) 1326). Because of the instability of the target of the antibody, which binds to tumor cells intermittently with only an intermittent mechanism, as described above, no anti-MUC 1 VNTR antibody has proven clinically effective yet. Wu et al, (CANCER CELL International) 22:417; fiedler, supra; kimura et al, (expert opinion on biological therapy) 13 (2013) 35-49; ibrahim et al, (CLIN CANCER RES) clinical cancer research 17 (2011) 6822-6830). In contrast to the alpha chain and its VNTR, the MUC1 SEA domain formed by the interaction of the alpha and beta subunits' extracellular portions is a stable, cell membrane-immobilized molecular moiety. anti-MUC 1 alpha/beta linked antibodies are disclosed in Rubenstein et al, cancer research CANCER RES, 66 (2006) 11247-11253; pichenak et al, cancer research 72 (2012) 3324-3336; rubenstein et al, international journal of Cancer (Int J Cancer), 124 (2009) 46-54. U.S. patent No.