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US-12624103-B2 - Engineered antibodies that bind LAG3

US12624103B2US 12624103 B2US12624103 B2US 12624103B2US-12624103-B2

Abstract

The present disclosure provides fully human anti-LAG3 IgG class antibodies engineered to have amino acid sequence in their heavy chain variable region and/or light chain variable region to improve antigen binding, cell binding, T cell activation and cytokine release capabilities.

Inventors

  • Damien Bresson
  • Min Soo Kim
  • Heyue Zhou
  • John Dixon Gray
  • Barbara A. Swanson
  • Alok Singh
  • Lisa Diane Kerwin

Assignees

  • YUHAN CORPORATION

Dates

Publication Date
20260512
Application Date
20210401

Claims (16)

  1. 1 . A LAG3-binding protein comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the CDR1, CDR2, and CDR3 sequences present in SEQ ID NO: 4 and the light chain variable region comprises the CDR1, CDR2, and CDR3 sequences present in SEQ ID NO: 5; or the heavy chain variable region comprises the CDR1, CDR2, and CDR3 sequences present in SEQ ID NO: 6 and the light chain variable region comprises the CDR1, CDR2, and CDR3 sequences present in SEQ ID NO:7; or the heavy chain variable region comprises the CDR1, CDR2, and CDR3 sequences present in SEQ ID NO: 8 and the light chain variable region comprises the CDR1, CDR2, and CDR3 sequences present in SEQ ID NO: 9; or the heavy chain variable region comprises the CDR1, CDR2, and CDR3 sequences present in SEQ ID NO: 10 and the light chain variable region comprises the CDR1, CDR2, and CDR3 sequences present in SEQ ID NO: 11; wherein the heavy chain region CDR1, CDR2, and CDR3 sequences, and the light chain variable region CDR1, CDR2, and CDR3 sequences are defined according to Kabat, Chothia, or IMGT numbering system, and optionally wherein the LAG3-binding protein is an antibody or an antigen-binding fragment thereof.
  2. 2 . The LAG3-binding protein of claim 1 , wherein the heavy chain variable region and the light chain variable region comprise the sequence of SEQ ID NOS: 4 and 5, SEQ ID NOS: 6 and 7, SEQ ID NOS: 8 and 9, or SEQ ID NOS: 10 and 11, respectively.
  3. 3 . The LAG3-binding protein of claim 1 , which is a fully human anti-LAG3 Fab fragment.
  4. 4 . The LAG3-binding protein of claim 1 , which is a single chain human anti-LAG3 antibody, and optionally wherein the variable domain region from a heavy chain and the variable domain region from a light chain are joined together with a peptide linker.
  5. 5 . The LAG3-binding protein of claim 4 , wherein the heavy chain variable region and the light chain variable region comprise the sequence of SEQ ID NOS: 4 and 5, SEQ ID NOS: 6 and 7, SEQ ID NOS: 8 and 9, or SEQ ID NOS: 10 and 11, respectively.
  6. 6 . The LAG3-binding protein of claim 1 , wherein the LAG3-binding protein binds human LAG3 with a K D of 10 −8 M or less; or wherein the LAG3-binding protein binds cynomolgus LAG3 with a K D of 10 −6 M or less.
  7. 7 . The LAG3-binding protein of claim 1 , comprising an IgG4 class antibody.
  8. 8 . A pharmaceutical composition comprising a pharmaceutically-acceptable excipient and the LAG3-binding protein of claim 1 .
  9. 9 . A kit comprising the LAG3-binding protein of claim 1 .
  10. 10 . A nucleic acid that encodes (a) (i) the heavy chain variable region of the LAG3-binding protein of claim 1 , and (ii) the light chain variable region of the LAG3-binding protein of claim 1 ; or (b) the LAG3-binding protein of claim 1 .
  11. 11 . A vector comprising the nucleic acid of claim 10 .
  12. 12 . A host cell comprising the vector of claim 11 , optionally wherein the vector comprises an expression vector, and wherein the host cell expresses the heavy and the light chain variable regions.
  13. 13 . A method for preparing a LAG3-binding protein, the method comprising: culturing a population of the host cell of claim 12 under conditions suitable for expressing the LAG3-binding protein, and optionally further comprising: recovering from the host cells the expressed LAG3-binding protein.
  14. 14 . A method of treating a disease associated with LAG3 over-expression in a subject in need thereof, comprising administering the LAG3-binding protein of claim 1 to the subject.
  15. 15 . The method of claim 14 , wherein the disease is bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, carcinoma of fallopian tubes, carcinoma of endometrium, carcinoma of cervix, carcinoma of vagina, carcinoma of vulva, Hodgkin's Disease, non-Hodgkin's lymphoma, cancer of esophagus, cancer of small intestine, cancer of endocrine system, cancer of thyroid gland, cancer of parathyroid gland, cancer of adrenal gland, sarcoma of soft tissue, cancer of urethra, cancer of penis, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, solid tumors of childhood, lymphocytic lymphoma, cancer of bladder, cancer of kidney or ureter, carcinoma of renal pelvis, neoplasm of central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancer, or a cancer induced by asbestos.
  16. 16 . The LAG3-binding protein of claim 2 , which is a fully human anti-LAG3 Fab fragment.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a national phase entry pursuant to 35 U.S.C. (371 of International Application No. PCT/US2021/025423, filed Apr. 1, 2021, which claims the benefit of priority of U.S. Provisional Patent Application No. 63/004,798, filed Apr. 3, 2020, each of which are incorporated by reference herein in their entireties for all purposes. Throughout this application various publications, patents, and/or patent applications are referenced. The disclosures of the publications, patents and/or patent applications are hereby incorporated by reference in their entireties into this application in order to more fully describe the state of the art to which this disclosure pertains. To the extent any material incorporated by reference conflicts with the express content of this application, the express content controls. 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 Jan. 14, 2021, is named 2021-01-14_01223-0075-00PCT_Seq_List_ST25.txt and is 23,793 bytes in size. TECHNICAL FIELD The present disclosure provides fully human anti-LAG3 IgG class antibodies engineered to have amino acid sequence in their heavy chain variable region and/or light chain variable region to improve antigen binding, cell binding, T cell activation and cytokine release capabilities. INTRODUCTION AND SUMMARY Lymphocyte Activation Gene-3, or LAG3 (also known as CD223), is a member of the immunoglobulin supergene family and is structurally and genetically related to CD4. LAG3 is not expressed on resting peripheral blood lymphocytes but is expressed on activated T cells and NK cells. LAG3 is a membrane protein encoded by a gene located on the distal part of the short arm of chromosome 12, near the CD4 gene, suggesting that the LAG3 gene may have evolved through gene duplication (Triebel et al. (1990) J. Exp. Med. 171:1393-1405). Similar to CD4, LAG3 has been demonstrated to interact with MHC Class II molecules but, unlike CD4, LAG3 does not interact with the human immunodeficiency virus gp120 protein (Baixeras et al. (1992) J. Exp. Med. 176:327-337). Studies using a soluble LAG3 immunoglobulin fusion protein (sLAG3Ig) demonstrated direct and specific binding of LAG3 to MHC class II on the cell surface (Huard et al. (1996) Eur. J Immunol. 26:1180-1186). In in vitro studies of antigen-specific T cell responses, the addition of anti-LAG3 antibodies led to increased T cell proliferation, higher expression of activation antigens such as CD25, and higher concentrations of cytokines such as interferon-gamma and interleukin-4, supporting a role for the LAG3/MHC class II interaction in down-regulating antigen-dependent stimulation of CD4+ T lymphocytes (Huard et al. (1994) Eur. J Immunol. 24:3216-3221). The intra-cytoplasmic region of LAG3 has been demonstrated to interact with a protein termed LAP, which is thought to be a signal transduction molecule involved in the downregulation of the CD3/TCR activation pathway (Iouzalen et al. (2001) Eur. J. Immunol. 31:2885-2891). Furthermore, CD4+CD25+ regulatory T cells (Treg) have been shown to express LAG3 upon activation and antibodies to LAG3 inhibit suppression by induced Treg cells, both in vitro and in vivo, suggesting that LAG3 contributes to the suppressor activity of Treg cells (Huang, C. et al. (2004) Immunity 21:503-513). Still further, LAG3 has been shown to negatively regulate T cell homeostasis by regulatory T cells in both T cell-dependent and independent mechanisms (Workman and Vignali (2005) J. Immunol. 174:688-695). In certain circumstances, LAG3 also has been shown to have immunostimulatory effects. For example, LAG3 transfected tumor cells transplanted into syngeneic mice showed growth reduction or complete regression as compared to untransfected tumor cells, suggesting that LAG3 expression on the tumor cells stimulated an anti-tumor response by triggering antigen LAG3 presenting cells via MHC class II molecules (Prigent et al. (1999) Eur. J. Immunol. 29:3867-3876). Additionally, soluble LAG3 Ig fusion protein has been shown to stimulate both humoral and cellular immune responses when administered to mice together with an antigen, indicating that soluble LAG3Ig can function as a vaccine adjuvant (El Mir and Triebel (2000) J. Immunol. 164:5583-5589). Furthermore, soluble human LAG3Ig has been shown to amplify in vitro generation of type I tumor-specific immunity (Casati et al. (2006) Cancer Res. 66:4450-4460). The functional activity of LAG3 is reviewed further in Triebel (2003) Trends Immunol. 24:619-622. In view of the above, additional agents for modulating the activity of LAG3 are of interest. There remains a need in the art for effective treatments based on LAG3, particularly anti-LAG3 antibodies. The present disclosure provides antibodies engineered to exhibit higher affinity binding to the