Search

CN-121986113-A - Humanized 40H3 antibodies

CN121986113ACN 121986113 ACN121986113 ACN 121986113ACN-121986113-A

Abstract

Antibodies (e.g., humanized monoclonal antibodies and antigen binding fragments) that specifically bind epidermal growth factor receptor variant III (egfrvlll) and/or gene-amplified EGFR, as well as conjugates and Chimeric Antigen Receptors (CARs) comprising such antibodies, are disclosed. Nucleic acid molecules encoding the antibodies, conjugates, or CARs disclosed herein, as well as host cells expressing these nucleic acid molecules, are also provided. Further disclosed are methods of using the disclosed compositions, e.g., for treating or detecting tumors, e.g., tumors that express egfrvlll and/or gene amplified EGFR.

Inventors

  • FITZGERALD DESMOND JAMES
  • A. Antignani
  • R. Sarnovsky

Assignees

  • 美国政府(由卫生和人类服务部的部长所代表)

Dates

Publication Date
20260505
Application Date
20240708
Priority Date
20230707

Claims (20)

  1. 1. An isolated humanized monoclonal antibody or antigen binding fragment thereof comprising a heavy chain variable region (V H ) and a light chain variable region (V L ), said V H and V L : a) Comprising SEQ ID NO 5 and SEQ ID NO 8 (A10, VH3+VL 3), respectively; b) Comprising SEQ ID NO 3 and SEQ ID NO 6 (A2, VH1+VL 1), respectively; c) Comprising SEQ ID NO 3 and SEQ ID NO 7 (A3, VH1+VL 2), respectively; d) Comprising SEQ ID NO 3 and SEQ ID NO 8 (A4, VH1+VL 3), respectively; e) Comprising SEQ ID NO 4 and SEQ ID NO 6 (A5, VH2+VL 1), respectively; f) Comprising SEQ ID NO 4 and SEQ ID NO 7 (A6, VH2+VL 2), respectively; g) Comprising SEQ ID NO 4 and SEQ ID NO 8 (A7, VH2+VL 3), respectively; h) Comprising SEQ ID NO 5 and SEQ ID NO 6 (A8, VH3+VL 1), respectively; i) Comprising SEQ ID NO 5 and SEQ ID NO 7 (A9, VH3+VL 2), respectively; j) Comprising SEQ ID NO: 1 and SEQ ID NO: 9 (B1, 40H3 VH+VL-EG), respectively; k) Comprising SEQ ID NO 1 and SEQ ID NO 10 (B2, 40H3 VH+VL-DA), respectively; l) comprises SEQ ID NO 4 and SEQ ID NO 11 (B3, VH2+VL 1-DA), respectively; m) comprises SEQ ID NO. 4 and SEQ ID NO. 12 (B4, VH2+VL 2-DA), respectively; n) comprises SEQ ID NO 5 and SEQ ID NO 11 (B5, VH3 and VL 1-DA), respectively; o) comprises SEQ ID NO. 31 and SEQ ID NO. 32 (C10), respectively; p) comprise SEQ ID NO 5 and SEQ ID NO 14 (D2), respectively, or Q) comprises SEQ ID NO 5 and SEQ ID NO 24 (D3), respectively.
  2. 2. The isolated humanized monoclonal antibody of claim 1, wherein the antibody comprises a human constant domain.
  3. 3. The isolated humanized monoclonal antibody of claim 1 or claim 2, wherein the antibody is IgG.
  4. 4. The isolated humanized monoclonal antibody of any one of the preceding claims, comprising a recombinant constant domain comprising a modification that extends the half-life of the antibody.
  5. 5. The isolated humanized monoclonal antibody or antigen binding fragment of any one of the preceding claims, conjugated to a toxin or chemotherapeutic agent.
  6. 6. The isolated humanized monoclonal antibody or antigen binding fragment of claim 5, wherein the toxin is pseudomonas (Pseudononas) exotoxin (PE), ricin, abrin, diphtheria toxin, ribosomal toxin, ribonuclease, saporin, calicheamicin, or botulinum toxin.
  7. 7. The isolated humanized monoclonal antibody or antigen binding fragment of claim 5 or claim 6, wherein the toxin is PE, and wherein the PE is PE25, PE38 or PE40.
  8. 8. The isolated humanized monoclonal antibody or antigen binding fragment of claim 5, wherein the chemotherapeutic agent is monomethyl auristatin E or maytansinoids.
  9. 9. The antigen binding fragment of any one of the preceding claims.
  10. 10. The antigen binding fragment of any one of the preceding claims, wherein the antigen binding fragment is Fv, dsFV, ds-scvFV, fab, F (ab') 2 , scFV, or scFV 2 fragment.
  11. 11. The antigen binding fragment of any one of the preceding claims, wherein the antigen binding fragment is a Fab or scFV.
  12. 12. The isolated humanized monoclonal antibody or antigen binding fragment of any one of the preceding claims, conjugated to a detectable marker.
  13. 13. A Chimeric Antigen Receptor (CAR) comprising the antigen binding fragment of any one of the preceding claims.
  14. 14. A bispecific antibody comprising the humanized monoclonal antibody or antigen-binding fragment of any one of claims 1-11.
  15. 15. An isolated nucleic acid molecule encoding the humanized antibody or antigen-binding fragment of any one of claims 1-12, or V H or V L of the antibody or antigen-binding fragment, the CAR of claim 13, or the bispecific antibody of claim 14.
  16. 16. The isolated nucleic acid molecule of claim 15, wherein the nucleic acid molecule is a cDNA sequence.
  17. 17. The isolated nucleic acid molecule of claim 15 or claim 16, operably linked to a promoter.
  18. 18. A vector comprising the nucleic acid molecule of any one of claims 15-17.
  19. 19. The vector of claim 18, wherein the vector is a viral vector.
  20. 20. An isolated host cell comprising the nucleic acid molecule of any one of claims 15-17, or the vector of claim 18 or 19.

Description

Humanized 40H3 antibodies Cross Reference to Related Applications The present application claims priority from U.S. provisional application No. 63/525,493 filed 7 at 2023, which is incorporated herein by reference in its entirety. Field of disclosure The present application relates to the field of cancer biology, in particular humanized monoclonal antibodies and antigen binding fragments, which specifically bind to epidermal growth factor receptor mutants/variants expressed by tumor cells (e.g., egfrvlll and/or gene amplified EGFR). Government support statement The present invention was completed under government support under item number z01#: z1abc 008757 from the national institute of health (National Institutes of Health) national institute of cancer (National Cancer Institute). The united states government has certain rights in this invention. Sequence listing reference The sequence listing is submitted with an XML file named "sequence. XML", which was created at month 7, 8 (1,386,226 bytes) of 2024, which is incorporated herein by reference. Background EGFR is often involved in the oncogenic progression of human cancers. Various changes in expression, including gene amplification and activating mutations, result in tumorigenesis. The human form of this large receptor has an extracellular domain of 621 amino acids (ECD), a single transmembrane domain of 23 amino acids (TM) and an enzymatically active intracellular domain of 542 amino acids (ICD). EGFR is a member of the receptor tyrosine kinase family and is the first receptor to show positive association with human cancers. Ligand binding results in receptor dimer formation and activation of the kinase domain, signaling one of several pathways that may promote mammalian cell growth, survival and diffusion. Activating mutations may occur in ECD or ICD. Gene amplification and deletion may also activate EGFR. Exemplary deletions include deletions of exons 2-7, which produce EGFR variant III (EGFRvIII), and deletions of exon 19 produce a constitutively active enzyme mutant. Gene amplification of EGFR, expression of egfrvlll or deletion of EGFR exon 19 is reported only in cancer cells. Antibodies typically bind tightly and specifically to their target antigens, making them useful therapeutic agents for the treatment of a wide variety of diseases characterized by altered protein expression (e.g., expression of EGFR by EGFRvIII or gene amplification). However, for many therapeutic applications, the efficacy and safety of non-human antibodies is compromised because non-human immunoglobulin (Ig) molecules are inherently immunogenic and thus capable of inducing anti-Ig immune responses, such that repeated dosing is highly compromised. Thus, for therapeutic use, it may be beneficial to modify an antibody to reduce or eliminate its immunogenicity. There remains a need for humanized antibodies that can bind egfrvlll or gene amplified EGFR but not wild-type EGFR to exhibit cancer cell specificity. Summary of the disclosure Isolated humanized monoclonal antibodies and antigen binding fragments thereof are disclosed, wherein the monoclonal antibodies specifically bind to EGFR in EGFRvIII and/or gene amplification. In some aspects, the monoclonal antibody or antigen-binding fragment comprises a heavy chain variable region (V H) and a light chain variable region (V L), V H and V L: a) Comprising SEQ ID NO 5 and SEQ ID NO 8 (A10, VH3+VL 3), respectively; b) Comprising SEQ ID NO 3 and SEQ ID NO 6 (A2, VH1+VL 1), respectively; c) Comprising SEQ ID NO 3 and SEQ ID NO 7 (A3, VH1+VL 2), respectively; d) Comprising SEQ ID NO 3 and SEQ ID NO 8 (A4, VH1+VL 3), respectively; e) Comprising SEQ ID NO 4 and SEQ ID NO 6 (A5, VH2+VL 1), respectively; f) Comprising SEQ ID NO 4 and SEQ ID NO 7 (A6, VH2+VL 2), respectively; g) Comprising SEQ ID NO 4 and SEQ ID NO 8 (A7, VH2+VL 3), respectively; h) Comprising SEQ ID NO 5 and SEQ ID NO 6 (A8, VH3+VL 1), respectively; i) Comprising SEQ ID NO 5 and SEQ ID NO 7 (A9, VH3+VL 2), respectively; j) Comprising SEQ ID NO: 1 and SEQ ID NO: 9 (B1, 40H3 VH+VL-EG), respectively; k) Comprising SEQ ID NO 1 and SEQ ID NO 10 (B2, 40H3 VH+VL-DA), respectively; l) comprises SEQ ID NO 4 and SEQ ID NO 11 (B3, VH2+VL 1-DA), respectively; m) comprises SEQ ID NO. 4 and SEQ ID NO. 12 (B4, VH2+VL 2-DA), respectively; n) comprises SEQ ID NO 5 and SEQ ID NO 11 (B5, VH3 and VL 1-DA), respectively; o) comprises SEQ ID NO. 31 and SEQ ID NO. 32 (C10), respectively; p) comprise SEQ ID NO 5 and SEQ ID NO 14 (D2), respectively, or Q) comprises SEQ ID NO 5 and SEQ ID NO 24 (D3), respectively. Conjugates of these antibodies and antigen binding fragments are also disclosed. Further disclosed are Chimeric Antigen Receptors (CARs), including the disclosed antibodies or antigen binding fragments, and cells (e.g., immune cells) expressing such chimeric antigen receptors (e.g., CAR T cells). Also disclosed are nucleic acid molecules encoding V H and/or V L of any of the antibodies