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CN-122003440-A - Anti-MUC 16 antibodies and uses thereof

CN122003440ACN 122003440 ACN122003440 ACN 122003440ACN-122003440-A

Abstract

The present disclosure provides MUC16 binding molecules, including anti-MUC 16 antibodies, particularly antibodies comprising VHH, methods of making and uses of the antibodies.

Inventors

  • WANG YIFENG
  • HU YAOHUA
  • WANG XIA
  • Z.WANG
  • NIE SIWEI
  • CHEN YUNYING
  • GU JIJIE

Assignees

  • 上海药明生物技术有限公司

Dates

Publication Date
20260508
Application Date
20240809
Priority Date
20230811

Claims (20)

  1. A muc16 binding molecule comprising an immunoglobulin single variable domain, wherein the single variable domain comprises CDR1, CDR2 and CDR3 of a VHH as set forth in SEQ ID No. 4 or 5.
  2. 2. The MUC 16-binding molecule of claim 1, wherein the CDR1 comprises the amino acid sequence shown in SEQ ID No. 1, the CDR2 comprises the amino acid sequence shown in SEQ ID No. 2, and the CDR3 comprises the amino acid sequence shown in SEQ ID No. 3.
  3. 3. The MUC 16-binding molecule of any one of claims 1 to 2, wherein the single variable domain comprises: (A) An amino acid sequence as shown in any one of SEQ ID NOs 4 to 5; (B) An amino acid sequence which is at least 85%, 90% or 95% identical to the amino acid sequence as set forth in any one of SEQ ID NOS.4-5, but which retains specific binding affinity for MUC 16.
  4. 4. The MUC 16-binding molecule of any one of claims 1 to 3, comprising one or more amino acid substitutions, additions and/or deletions in a framework region of the single variable domain, e.g., FRW1, FRW2, FRW3 and/or FRW 4.
  5. 5. The MUC 16-binding molecule of any one of claims 1 to 6, wherein the single variable domain comprises the amino acid sequence set forth in SEQ ID No. 4 or 5.
  6. 6. The MUC 16-binding molecule of any one of claims 1 to 5, wherein the MUC 16-binding molecule further comprises a human IgG constant domain.
  7. 7. The MUC 16-binding molecule of claim 6, wherein the human IgG constant domain is a human IgG1, igG2, igG3 or IgG4 constant domain, such as a human IgG1 constant domain or variant thereof.
  8. 8. The MUC 16-binding molecule of any one of claims 1 to 7, having one or more of the following properties: (a) Specifically bind to human MUC16 and cynomolgus MUC16 but not CA125 soluble proteins; (b) Has a low tendency to self-interaction; (c) Has good thermal stability, and (D) There is no non-specific binding.
  9. 9. The MUC 16-binding molecule of any one of claims 1 to 8, wherein the MUC 16-binding molecule is a chimeric or humanized antibody.
  10. 10. The MUC16 binding molecule of any one of claims 1 to 9, comprising an amino acid sequence as set forth in any one of SEQ ID NOs 6-7.
  11. 11. The MUC16 binding molecule of any one of claims 1 to 10, which is a dimer.
  12. 12. A fusion protein comprising a MUC16 binding molecule as defined in any one of claims 1 to 11 fused to a heterologous peptide, such as an antigen binding domain targeting a different antigen.
  13. 13. A nucleic acid molecule comprising a nucleic acid sequence encoding a single variable domain of a MUC16 binding molecule as defined in any one of claims 1 to 12.
  14. 14. A vector comprising the nucleic acid molecule of claim 13.
  15. 15. A host cell comprising the vector of claim 16 or the nucleic acid molecule of claim 13.
  16. 16. A pharmaceutical composition comprising at least one MUC16 binding molecule as defined in any one of claims 1 to 11 and a pharmaceutically acceptable carrier.
  17. 17. A method for producing a MUC16 binding molecule comprising the steps of: -culturing the host cell of claim 15 under conditions suitable for expression of said MUC16 binding molecule, and -Isolating the MUC16 binding molecules from the culture supernatant.
  18. 18. A method of modulating a MUC 16-associated immune response in a subject, comprising administering to the subject a MUC 16-binding molecule as defined in any one of claims 1 to 11 or a pharmaceutical composition of claim 16, such that the immune response is modulated in the subject.
  19. 19. A method for treating or preventing cancer in a subject, comprising administering to the subject an effective amount of a MUC16 binding molecule as defined in any one of claims 1 to 11 or a pharmaceutical composition of claim 16, wherein the cancer is MUC16 positive or overexpressed.
  20. 20. The method of claim 19, wherein the cancer is selected from ovarian cancer, lung cancer, pancreatic cancer, breast cancer, uterine cancer, fallopian tube cancer, primary peritoneal cancer, or cancer of any other tissue expressing MUC 16.

Description

Anti-MUC 16 antibodies and uses thereof Sequence listing The present application comprises a sequence listing, which is incorporated herein by reference in its entirety. Technical Field The present application relates generally to antibodies. More specifically, the application relates to monoclonal antibodies against MUC16, in particular antibodies comprising VHH, methods for their preparation and uses of the antibodies. Background Mucin 16 (MUC 16, previously known as cancer antigen 125, CA 125) was first identified by Bast et al in 1981, and its cDNA sequence was later found to correspond to mucin MUC16 [1-2]. MUC16 is a highly glycosylated single-transmembrane protein with a molecular weight of about 3000-5000 kD [2]. It is the largest mucin, consisting of a number of domains, including the extracellular N-terminal domain, the large tandem repeat domain interspersed with SEA urchin sperm, enterokinase, and collectin (SEA) domains, and the C-terminal domain that contains a transmembrane region and a short cytoplasmic tail [3]. MUC16 has 56 SEA domains, with the penultimate SEA (55 th) domain having a conserved cleavage site [4]. MUC16 can be shed from the cell surface and released into the blood stream, proteolytically cleaving into soluble MUC16 (CA 125), and the extracellular domain that remains between the cell membrane and the cleavage site remains on the cell surface [4]. Under normal physiological conditions, MUC16 is expressed at low levels in only a few tissues, including the respiratory tract and female genital tract, particularly in glandular and epithelial cells [5]. MUC16 is expressed at significantly higher levels than normal tissue in a range of human cancers including ovarian, endometrial, and pancreatic cancers. The shed domain, called CA125, is a poor prognostic and diagnostic serum marker for ovarian cancer. CA125 is the most widely used ovarian tumor marker and is generally considered to be the "gold standard". [6, 7]. Abnormal CA125 levels (> 35U/mL) were observed in 99% in serous cancer patients graded from grade I to IV in the FIGO (international union of obstetrics and gynecology (International Federation of Gynecologists and Obstetricians)). In many FIGO Iv serous ovarian cancer patients, serum CA125 levels can be increased up to (up to) 10-fold and over 2000U/mL [8] as compared to stage I. In addition to ovarian cancer, elevated expression of CA125 is strongly correlated with poor prognosis for a variety of cancers [9]. The limited expression of MUC16 on normal human tissue and its high expression in many common cancers makes it an attractive target for cancer therapies. Although several therapeutic antibodies targeting MUC16, including ago Fu Shan antibody (oregovomab) and aba Fu Shan antibody (abagovomab), have been tested in clinical trials, only limited efficacy has been achieved in cancer patients [10, 11]. A potential disadvantage of therapeutic agents based on several of the described antibodies is that they target the membrane distal region of MUC16, and therefore, due to the high levels of circulating CA125 and CA125 antigen sinking effects (SINK EFFECT) in cancer patients, the antibodies bound by the target cells are significantly reduced, and the tumor killing effects will therefore be impaired [4]. In developing therapeutic antibodies that target MUC16 positive cancers, it may be critical to avoid binding to soluble CA125 in the blood circulation. There remains a need to develop new anti-MUC 16 antibodies, preferably those that bind to MUC16 extracellular domains on the cell membrane, rather than shed CA125, to minimize the antigen sinking effects from high levels of soluble CA125 in cancer patients. Disclosure of Invention The present disclosure relates to compounds, methods, compositions and articles of manufacture that provide MUC16 binding molecules with improved efficacy. The benefits provided by the present disclosure are broadly applicable to the field of antibody therapeutics and diagnostics, and can be used in combination with other therapeutics (such as antibodies that react with multiple targets). The present disclosure provides MUC16 binding molecules, such as monoclonal antibodies, that can specifically bind to human MUC16 and cross-react with cynomolgus MUC 16. Such MUC16 binding molecules provide certain advantages over reagents, compositions and/or methods currently used and/or known in the art. These advantages include improved therapeutic and pharmacological properties, increased specificity, reduced immunogenicity, and other advantageous properties. In the present disclosure, MUC16 binding molecules, such as monoclonal antibodies, to MUC16 have been developed that can be used to treat tumors that overexpress MUC 16. The present disclosure provides MUC16 binding molecules, nucleic acid molecules encoding the same, expression vectors and host cells for expressing the MUC16 binding molecules, and methods of using the MUC16 binding molecules. The MUC16 b