Antibody molecules against LAG-3 and their use

Abstract

To provide novel agents that modulate the activity of the immune system to activate it, considering the importance of LAG-3 in downregulating an immune response.SOLUTION: Antibody molecules that specifically bind to LAG-3 are disclosed. The anti-LAG-3 antibody molecules can be used to treat, prevent and / or diagnose cancerous or infectious disorders.SELECTED DRAWING: Figure 5

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority under U.S. Provisional Application No. 61 / 953,536 (filed March 14, 2014), U.S. Provisional Application No. 62 / 059,690 (filed October 3, 2014), and U.S. Provisional Application No. 62 / 094,889 (filed December 19, 2014), and incorporates the contents of the said applications in their entirety by reference.

[0002] Sequence List This application includes a sequence listing, which has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy was created on 10 March 2015, is named C2160-7001WO_SL.txt, and is 258,890 bytes in size. [Background technology]

[0003] background Lymphocyte-activating gene-3, or LAG-3 (also known as CD223), is a member of the immunoglobulin supergene family and is expressed in activated T cells (Huard et al. (1994) Immunogenetics 39:213), NK cells (Triebel et al. (1990) J. Exp. Med. 171:1393-1405), regulatory T cells (Huang et al. (2004) Immunity 21:503-513; Camisaschi et al. (2010) J Immunol. 184:6545-6551; Gagliani et al. (2013) Nat Med 19:739-746), and plasmacytoid dendritic cells (DCs) (Workman et al. (2009) J Immunol 182:1885-1891). LAG-3 is a membrane protein encoded by a gene located on chromosome 12 and is structurally and genetically related to CD4.

[0004] Similar to CD4, LAG-3 can interact with MHC class II molecules on the cell surface (Baixeras et al. (1992) J. Exp. Med. 176:327-337; Huard et al. (1996) Eur. J. Immunol. 26:1180-1186). Direct binding of LAG-3 to MHC class II is similar to that of CD4. + It has been suggested that LAG-3 plays a role in downregulating antigen-dependent stimulation of T lymphocytes (Huard et al. (1994) Eur. J. Immunol. 24:3216-3221), and LAG-3 blockade is also effective in both tumor or autoantigen (Gross et al. (2007) J Clin Invest. 117:3383-3392) and viral models (Blackburn et al. (2009) Nat. Immunol. 10:29-37) for CD8 + It has also been shown to reactivate lymphocytes. Furthermore, the cytoplasmic domain of LAG-3 can interact with LAP (LAG-3-related protein), a signaling molecule involved in the downregulation of the CD3 / TCR activation pathway (Iouzalen et al. (2001) Eur. J. Immunol. 31:2885-2891). In addition, CD4 + CD25 + Regulatory T cells (T reg It has been shown that ) expresses LAG-3 upon activation, which is T reg It contributes to the suppressor activity of cells (Huang, C. et al. (2004) Immunity 21:503-513). LAG-3 also acts on T cells through both T cell-dependent and T cell-independent mechanisms. reg Cells can also negatively regulate T cell homeostasis (Workman, CJ and Vignali, DA (2005) J. Immunol. 174:688-695). [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Huard et al. (1994) Immunogenetics 39:213 [Non-licensed document 2] Triebel et al. (1990) J. Exp. Med. 171:1393-1405 [Non-licensed document 3] Huang et al. (2004) Immunity 21:503-513

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Summary of the Invention

Problems to be Solved by the Invention

[0006] Considering the importance of LAG-3 in the downregulation of the immune response, there is a need for new agents that modulate its activity to activate the immune system. Such agents can be used, for example, in the treatment of other conditions such as cancer immunotherapy and chronic infections.

Means for Solving the Problems

[0007] Summary Disclosed herein are antibody molecules (e.g., humanized antibody molecules) that bind to lymphocyte activation gene-3 (LAG-3) with high affinity and specificity. In certain embodiments, the anti-LAG-3 antibody molecules comprise novel combinations of framework regions (e.g., FW1, FW2, FW3 and / or FW4), e.g., novel combinations of heavy chain framework regions and / or light chain framework regions. Nucleic acid molecules, expression vectors, host cells and methods for producing the antibody molecules are also provided. Immune complexes, multi- or bispecific antibody molecules and pharmaceutical compositions comprising the antibody molecules are also provided. The anti-LAG-3 antibody molecules disclosed herein can be used (alone or in combination with other agents or therapies) in the treatment, prevention and / or diagnosis of cancerous disorders (e.g., solid and soft tissue tumors) as well as infectious diseases. Accordingly, compositions and methods for detecting LAG-3 and methods for treating a variety of disorders including cancer and / or infectious diseases using anti-LAG-3 antibody molecules are disclosed herein.

[0008] Thus, in one aspect, the invention relates to antibody molecules (e.g., isolated or recombinant antibody molecules) having one or more of the following characteristics. (i) High affinity for LAG-3, e.g., human LAG-3, e.g., at least about 10 7 M -1Generally about 10 8 M -1 , more generally about 10 9 M -1 ~10 10 M -1 They bind with the above affinity constants; (ii) LAG-3, for example, LAG-3-CHO transimplants, with K content of 5nM, 4nM, 3nM, 2nM, 1nM, for example, less than 1-3nM (e.g., about 1.92nM or about 2.3nM) D Join them together; (iii) Does not bind substantially to CD4; (iv) Inhibits the binding of LAG-3 to major histocompatibility (MHC) class II molecules, for example, approximately 1-20 nM, 5-15 nM, for example, 5.5 nM IC 50 To show; (v) Binding to the D1 domain of LAG-3 (e.g., human LAG-3), for example, binding to the D1 domain but not to the extra loop region of the D1 domain; (vi) modulate (e.g., stimulate, enhance, or restore) an immune response, such as an antigen-specific T cell response or an antitumor response; (vii) Specifically binds to epitopes identical or similar to LAG-3 epitopes, for example, epitopes recognized by the mouse monoclonal antibody BAP050 or the chimeric antibody BAP050-chi; (viii) It binds to an epitope on LAG-3 that is different from the one recognized by antibody BMS-986016; (ix)BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP05 0-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BA P050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20 , huBAP050(Ser) (e.g. BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, B AP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP 050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050 -hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I or BAP050-Clone-J exhibit the same or similar binding affinity or specificity, or both;

[0009] (x) Exhibits the same or similar binding affinity or specificity, or both, to the antibody molecule (e.g., heavy chain variable region and light chain variable region) listed in Table 1; (xi) Exhibits the same or similar binding affinity or specificity, or both, to an antibody molecule having the amino acid sequence shown in Table 1 (e.g., heavy chain variable region and light chain variable region); (xii) Exhibiting the same or similar binding affinity or specificity, or both, to the antibody molecule encoded by the nucleotide sequence shown in Table 1 (e.g., heavy chain variable region and light chain variable region); (xiii) Inhibit the binding of the second antibody molecule to LAG-3, for example, competitively inhibit, where the second antibody molecule is the antibody molecule described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP0 50-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BAP050-hum15, BAP050-hum16, BAP050- hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g., BAP050-hum01-Ser, BAP050-hum02-Se r, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050- hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, A molecule selected from BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I or BAP050-Clone-J, for example, an antibody molecule; (xiv) Binds to LAG-3 at an epitope identical or overlapping with that of the second antibody molecule, where the second antibody molecule is one of the antibody molecules listed herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP0 50-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BAP050-hum15, BAP050-hum16, BAP050- hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g., BAP050-hum01-Ser, BAP050-hum02-Se r, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050- hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, A molecule selected from BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I or BAP050-Clone-J, for example, an antibody molecule; (xv) For example, measured by Biacore, FACS, or both, to determine whether the second antibody molecule competes with and / or binds to the same epitope as the second antibody molecule, where the second antibody molecule is one of the antibody molecules described herein, e.g., BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BAP050-h um15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (for example, BAP050-hum01 -Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hu m07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050- An antibody molecule selected from one of the following: hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I or BAP050-Clone-J;

[0010] (xvi) The antibody molecules described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP 050-hum13, BAP050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BA P050-hum20, huBAP050(Ser) (e.g. BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum 04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Se r, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP A selected antibody molecule having one or more biological properties, for example, from among 050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I or BAP050-Clone-J; (xvii) The antibody molecules described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP 050-hum13, BAP050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BA P050-hum20, huBAP050(Ser) (e.g. BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum0 4-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser , BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP05 A selected antibody molecule having one or more pharmacokinetic properties, for example, from among 0-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I or BAP050-Clone-J; or (xviii) Inhibiting one or more activities of LAG-3, for example, CD4 + Increased antigen-dependent stimulation of T lymphocytes; increased T cell proliferation; increased expression of activating antigens, e.g., CD25; increased expression of cytokines, e.g., interferon-gamma (IFN-γ), interleukin-2 (IL-2), or interleukin-4 (IL-4); increased expression of chemokines, e.g., CCL3, CCL4, or CCL5; T regThis results in one or more of the following: decreased cellular suppressor activity; increased T cell homeostasis; increased tumor-infiltrating lymphocytes; or decreased immune evasion by cancer cells.

[0011] The term "huBAP050(Ser)" used herein refers to any humanized BAP050 antibody molecule having a Cys-to-Ser substitution at position 84 of the heavy chain framework region 3 (VHFW3), such as those listed in Table 1, or any of the humanized BAP050 antibody molecules listed herein. In one embodiment, the huBAP050(Ser) antibody molecule is BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050- Select from hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser, or BAP050-hum20-Ser.

[0012] In one embodiment, the anti-LAG-3 antibody molecule has high affinity for LAG-3, for example, a mouse or chimeric anti-LAG-3 antibody molecule, for example, the dissociation equilibrium constant (K) of the mouse or chimeric anti-LAG-3 antibody molecule described herein. D ) is approximately the same as or at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% higher or lower K D It binds to LAG-3, for example, LAG-3-CHO transtransferant, with K2 levels of 5nM, 4nM, 3nM, 2nM, for example, 1-3nM (e.g., about 1.92nM or about 2.3nM). D They are joined together.

[0013] In one embodiment, the expression level of the anti-LAG-3 antibody molecule is approximately the same as, higher than, or lower than, the expression level of a mouse or chimeric antibody molecule, e.g., the mouse or chimeric anti-LAG-3 antibody molecule described herein, e.g., at least about 0.5 times, 1 time, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, or 10 times higher or lower. In one embodiment, the antibody molecule is expressed in CHO cells.

[0014] In one embodiment, an anti-LAG-3 antibody molecule exhibits one or more LAG-3-related activities, such as the IC of a mouse or chimeric anti-LAG-3 antibody molecule, e.g., the mouse or chimeric anti-LAG-3 antibody molecule described herein. 50 Approximately the same as (50% inhibitory concentration), higher or lower, for example, at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% higher or lower IC. 50 Inhibits LAG-3 activity. In one embodiment, LAG-3-related activity is the binding of MHC class II molecules to LAG-3. In another embodiment, LAG-3-related activity is the binding of L-SECtin to LAG-3. In one embodiment, anti-LAG-3 antibodies inhibit IC50 at approximately 1-20 nM, 5-15 nM, and 5.5 nM. 50 It has (for example, detectable by inhibition of MHC class II or L-SECtin binding).

[0015] In one embodiment, the anti-LAG-3 antibody molecule has substantially the same or improved stability as the mouse or chimeric anti-LAG-3 antibody molecule, e.g., the mouse or chimeric anti-LAG-3 antibody molecule described herein, e.g., it is at least about 0.5-fold, 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or 10-fold stable in vivo or in vitro.

[0016] In one embodiment, the anti-LAG-3 antibody molecule is a humanized antibody molecule having a risk score of 800-1200, 850-1150, 900-1100, 950-1050, or any of the risk scores listed herein, based on T cell epitope analysis.

[0017] In other aspects, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BA P050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g., BAP050-hum01-Se r, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050 -hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14- Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP05 The antibody comprises at least one antigen-binding region, e.g., a variable region or its antigen-binding fragment, from an antibody selected from 0-Clone-H, BAP050-Clone-I, or BAP050-Clone-J; or one encoded by the nucleotide sequence as described in Table 1 or in Table 1; or a sequence substantially identical to any of the aforementioned sequences (e.g., matching at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, or higher).

[0018] In another aspect, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-h um14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g., BAP050 -hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07 -Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP05 An antibody selected from any of 0-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J; or one encoded by the nucleotide sequences listed in Table 1 or in Table 1; or comprising at least 1, 2, 3, or 4 variable regions from a sequence that is substantially identical to any of the aforementioned sequences (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher matching). In one embodiment, the antibody molecule includes a substitution in the heavy chain framework region 3 (VHFW3) (e.g., a Cys-to-Ser substitution at position 84) (e.g., as shown in Tables 1 and 2).

[0019] In another aspect, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050- hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (for example, BAP05 0-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum0 7-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser , BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP0 An antibody selected from any of 50-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J; or one encoded by the nucleotide sequence as described in Table 1 or in Table 1; or one comprising at least one or two-stranded variable regions from a sequence substantially identical to any of the aforementioned sequences (e.g., matching at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, or higher).

[0020] In another aspect, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050- hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (for example, BAP05 0-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum0 7-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser , BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP0 An antibody selected from any of 50-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J; or one encoded by the nucleotide sequence as described in Table 1 or in Table 1; or comprising at least one or two light chain variable regions from a sequence that is substantially identical to any of the aforementioned sequences (e.g., matching at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, or higher).

[0021] In yet another embodiment, the anti-LAG-3 antibody molecule comprises the heavy chain constant region of IgG4, e.g., human IgG4. In one embodiment, human IgG4 includes a substitution at EU numbering position 228 (e.g., Ser to Pro substitution). In yet another embodiment, the anti-LAG-3 antibody molecule comprises the heavy chain constant region of IgG1, e.g., human IgG1. In one embodiment, human IgG1 includes a substitution at EU numbering position 297 (e.g., Asn to Ala substitution). In one embodiment, human IgG1 includes a substitution at EU numbering position 265, a substitution at EU numbering position 329, or both (e.g., Asp to Ala substitution at EU numbering position 265 and / or Pro to Ala substitution at EU numbering position 329). In one embodiment, human IgG1 includes a substitution at EU numbering position 234, a substitution at EU numbering position 235, or both (e.g., Leu to Ala substitution at EU numbering position 234 and / or Leu to Ala substitution at EU numbering position 235). In one embodiment, the heavy chain constant region includes a sequence that is substantially identical to the amino acid sequence shown in Table 3 (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher).

[0022] In yet another embodiment, the anti-LAG-3 antibody molecule includes a kappa light chain constant region, for example, a human kappa light chain constant region. In one embodiment, the light chain constant region includes an amino acid sequence shown in Table 3 or a sequence substantially identical thereto (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, or higher).

[0023] In other embodiments, the anti-LAG-3 antibody molecule comprises IgG4, e.g., the heavy chain constant region and the kappa light chain constant region of human IgG4, e.g., the human kappa light chain constant region, and includes the heavy chain and light chain constant regions containing sequences that are substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher) to the amino acid sequences shown in Table 3. In one embodiment, the constant region is mutant IgG4, e.g., mutant human IgG4 (e.g., having a mutation at position 228 according to EU numbering (e.g., the S228P mutation)). In yet another embodiment, the anti-LAG-3 antibody molecule includes the heavy chain constant region and the kappa light chain constant region of IgG1, e.g., human IgG1, e.g., the human kappa light chain constant region, e.g., the amino acid sequence shown in Table 3 or substantially identical thereto (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher). In one embodiment, human IgG1 includes a substitution at EU numbering position 297 (e.g., Asn to Ala substitution). In another embodiment, human IgG1 includes a substitution at EU numbering position 265, a substitution at EU numbering position 329, or both (e.g., Asp to Ala substitution at EU numbering position 265 and / or Pro to Ala substitution at EU numbering position 329). In one embodiment, human IgG1 includes substitution at EU numbering position 234, substitution at EU numbering position 235, or both (e.g., Leu to Ala substitution at EU numbering position 234 and / or Leu to Ala substitution at EU numbering position 235).

[0024] In other aspects, the anti-LAG-3 antibody molecules are BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BAP050-hum 15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g., BAP050-hum01-Ser, BAP0 50-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hu m08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-S er, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP The anti-LAG-3 antibody molecule may optionally include a heavy chain variable domain and constant region, a light chain variable domain and constant region, or both, containing the amino acid sequence of 050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J; or encoded by the nucleotide sequence as described in Table 1 or in Table 1; or a sequence substantially identical to any of the aforementioned sequences (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, or higher). The anti-LAG-3 antibody molecule may optionally include a leader sequence from the heavy chain, light chain, or both, as shown in Table 4; or a sequence substantially identical thereto.

[0025] In another aspect, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BAP050-hum15, BAP050- hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g. BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum 03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10 An antibody selected from one of the following: -Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I or BAP050-Clone-J; or Table 1 The sequences described in; or encoded by the nucleotide sequences in Table 1; or sequences substantially identical to any of the aforementioned sequences (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher matching); or comprising at least 1, 2, or 3 complementarity-determining regions (CDRs) from the heavy chain variable region of a sequence having at least one amino acid change compared to the 1, 2, or 3 CDRs shown in Table 1, but not more than 2, 3, or 4 changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions).

[0026] In one embodiment, the anti-LAG-3 antibody molecule comprises at least one, two, or three CDRs (or collectively, all CDRs) from a heavy chain variable region containing or encoded by the amino acid sequences shown in Table 1. In one embodiment, one or more CDRs (or collectively, all CDRs) have one, two, three, four, five, six, or more changes, e.g., amino acid substitutions or deletions, compared to the amino acid sequences shown in or encoded by the nucleotide sequences shown in Table 1.

[0027] In another aspect, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BAP050-hum15, BAP050- hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g. BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum 03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10 An antibody selected from one of the following: -Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I or BAP050-Clone-J; or Table 1 The sequences described in; or encoded by the nucleotide sequences in Table 1; or sequences that are substantially identical to any of the aforementioned sequences (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher matching); or sequences comprising at least 1, 2, or 3 complementarity-determining regions (CDRs) from the light chain variable region of a sequence having at least one amino acid change compared to the 1, 2, or 3 CDRs shown in Table 1, but not more than 2, 3, or 4 changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions).

[0028] In yet another embodiment, the anti-LAG-3 antibody molecule comprises at least one, two, or three CDRs (or collectively all CDRs) from the light chain variable region containing or encoded by the amino acid sequences shown in Table 1. In one embodiment, one or more CDRs (or collectively all CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions or deletions, compared to the amino acid sequences shown in or encoded by the nucleotide sequences shown in Table 1.

[0029] In another embodiment, the anti-LAG-3 antibody molecule comprises at least 1, 2, 3, 4, 5, or 6 CDRs (or collectively, all CDRs) from the heavy and light chain variable regions containing or encoded by the amino acid sequences shown in Table 1. In one embodiment, one or more CDRs (or collectively, all CDRs) are amino acid sequences shown in or encoded by the nucleotide sequences shown in Table 1; or sequences that are substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher) to any of the aforementioned sequences; or sequences that have at least one amino acid change compared to the 1, 2, 3, 4, 5, or 6 CDRs shown in Table 1, but not more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions), and have one, two, three, four, five, six, or more changes, e.g., amino acid substitutions or deletions.

[0030] In another embodiment, the anti-LAG-3 antibody molecule comprises at least 1, 2, 3, 4, 5, or 6 CDRs (or collectively, all CDRs) from the heavy chain and the light chain variable region which includes or is encoded by the amino acid sequences shown in Table 1. In one embodiment, one or more CDRs (or collectively, all CDRs) have 1, 2, 3, 4, 5, 6, or more changes, e.g., amino acid substitutions or deletions, compared to the amino acid sequences shown in or encoded by the nucleotide sequences shown in Table 1.

[0031] In one respect, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BAP0 50-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g., BAP050-hum01-Ser, B AP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum 08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, B AP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clon An antibody selected from e-H, BAP050-Clone-I, or BAP050-Clone-J; or a total of 6 CDRs from CDRs that are identical to or have at least one amino acid change compared to CDRs 1, 2, 3, 4, 5, or 6 shown in Table 1, but have not been changed beyond 2, 3, or 4 (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions). In one embodiment, an anti-LAG-3 antibody molecule may contain any of the CDRs described herein.

[0032] In one aspect, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP 050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (for example For example, BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BA P050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), B An antibody selected from AP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J, or one listed in Table 1; or one encoded by a nucleotide sequence in Table 1; or a sequence substantially identical to any of the aforementioned sequences (e.g., matching at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, or higher);Alternatively, it includes at least 1, 2, or 3 CDRs by Kabat (e.g., at least 1, 2, or 3 CDRs by Kabat as defined in Table 1) from the heavy chain variable region, having at least one amino acid change but not more than 2, 3, or 4 changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions), compared to at least 1, 2, or 3 CDRs by Kabat as shown in Table 1.

[0033] In one respect, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP0 50-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g. , BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, B AP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP 050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BA An antibody selected from P050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J; or one listed in Table 1; or one encoded by a nucleotide sequence in Table 1; or a sequence substantially identical to any of the aforementioned sequences (e.g., matching at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, or higher);Alternatively, it includes at least 1, 2, or 3 CDRs by Kabat (e.g., at least 1, 2, or 3 CDRs by Kabat as defined in Table 1) from the light chain variable region that have at least one amino acid change, but not more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions), compared to at least 1, 2, or 3 CDRs by Kabat as shown in Table 1.

[0034] In another aspect, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, B AP050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser)( For example, BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser , BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, B AP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), B An antibody selected from AP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J; or one listed in Table 1; or one encoded by a nucleotide sequence in Table 1; or a sequence substantially identical to any of the aforementioned sequences (e.g., matching at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, or higher);Alternatively, it may include at least 1, 2, 3, 4, 5, or 6 CDRs from the heavy and light chain variable regions of Kabat et al. (e.g., at least 1, 2, 3, 4, 5, or 6 CDRs according to Kabat's definition as shown in Table 1) that have at least one amino acid change but not more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions), compared to at least 1, 2, 3, 4, 5, or 6 CDRs by Kabat et al. shown in Table 1. In some embodiments, an anti-LAG-3 antibody molecule may contain any of the CDRs described herein.

[0035] In another aspect, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BAP050-hum15, BAP050-hum m16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g., BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03- Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser An antibody selected from BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I or BAP050-Clone-J; or one of those listed in Table 1. ; or those encoded by the nucleotide sequences in Table 1; or sequences substantially identical to any of the aforementioned sequences (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher matching); or including all 6 CDRs by Kabat from the heavy and light chain variable regions (e.g., all 6 CDRs by Kabat definition as shown in Table 1) that have at least one amino acid change but not more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions).In one embodiment, the anti-LAG-3 antibody molecule may include any of the CDRs described herein.

[0036] In other aspects, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, B AP050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser ) (for example, BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06 -Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12 -Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum2 An antibody selected from any of the following: 0-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J; or one encoded by a nucleotide sequence in Table 1; or a sequence substantially identical to any of the aforementioned sequences (e.g., matching at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, or higher);Alternatively, it includes at least one, two, or three highly variable loops from the heavy chain variable region that have at least one amino acid change, but not more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions), compared to Chothia's definition of one, two, or three highly variable loops as shown in Table 1.

[0037] In other aspects, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, B AP050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser ) (for example, BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06 -Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12 -Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum2 An antibody selected from any of the following: 0-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J; or one encoded by a nucleotide sequence in Table 1; or a sequence substantially identical to any of the aforementioned sequences (e.g., matching at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, or higher);Alternatively, compared to Chothia's 1, 2, or 3 highly variable loops as shown in Table 1, this includes Chothia's 1, 2, or 3 highly variable loops (e.g., at least 1, 2, or 3 CDRs as defined by Chothia, as shown in Table 1) from the light chain variable region having at least one amino acid change but not more than 2, 3, or 4 changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions).

[0038] In another aspect, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, B AP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (for example, BAP050-hum01-Se r, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050- hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Se r, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-C An antibody selected from lone-H, BAP050-Clone-I, or BAP050-Clone-J; or one listed in Table 1; or one encoded by the nucleotide sequence in Table 1; or comprising at least 1, 2, 3, 4, 5, or 6 highly variable loops (e.g., at least 1, 2, 3, 4, 5, or 6 highly variable loops by Chothia definition as shown in Table 1) from the heavy and light chain variable regions of amino acids from at least one highly variable loop in contact with LAG-3. In one embodiment, the anti-LAG-3 antibody molecule comprises at least 1, 2, 3, 4, 5, or 6 Chothia highly variable loops in Table 1.

[0039] In one aspect, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BA P050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g., BAP050-hum01-Se r, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050 -hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14- Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP05 The antibody selected from either 0-Clone-H, BAP050-Clone-I, or BAP050-Clone-J, or a closely related highly variable loop, for example, all six highly variable loops (e.g., all six highly variable loops by Chothia definition as shown in Table 1) that are identical to or have at least one amino acid change but not more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions). In some embodiments, an anti-LAG-3 antibody molecule may contain any of the highly variable loops described herein.

[0040] In another aspect, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum1 3, BAP050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(S er) (e.g. BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum0 6-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12 -Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20 It contains at least one, two, or three highly variable loops having the same standard structure as the corresponding highly variable loop of an antibody selected from any of the following: BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J, for example, at least one loop 1 and / or loop 2 of the heavy chain and / or light chain variable domain of the antibody described herein. For example, see Chothia et al., (1992) J. Mol. Biol. 227:799-817; Tomlinson et al., (1992) J. Mol. Biol. 227:776-798 for a description of the highly variable loop standard structure. These structures can be determined by examining the tables described in these references.

[0041] In one embodiment, the anti-LAG-3 antibody molecule comprises a combination of CDR or highly variable loop as defined by Kabat et al. and Chothia et al.

[0042] In one aspect, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, B AP050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser ) (for example, BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06 -Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12 -Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum2 An antibody selected from any of the following: 0-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J; or one encoded by a nucleotide sequence in Table 1; or a sequence substantially identical to any of the aforementioned sequences (e.g., matching at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, or higher);Alternatively, it includes at least 1, 2, or 3 CDRs or highly variable loops as defined by Kabat and Chothia (e.g., at least 1, 2, or 3 CDRs or highly variable loops as defined by Kabat and Chothia, as shown in Table 1), which have at least one amino acid change but not more than 2, 3, or 4 changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) from the heavy chain variable region.

[0043] For example, an anti-LAG-3 antibody molecule may include, for example, Kabat et al.'s VH CDR1 or Chothia et al.'s VH highly variable loop 1 or a combination thereof, as shown in Table 1. In one embodiment, the Kabat and Chothia CDR combination of VH CDR1 includes the amino acid sequence GFTLTNYGMN (SEQ ID NO: 286) or a substantially identical amino acid sequence (e.g., having at least one amino acid change but not more than two, three, or four changes (e.g., substitution, deletion, or insertion, e.g., conservative substitution)). The anti-LAG-3 antibody molecule may further include, for example, Kabat et al.'s VH CDR2-3 and Kabat et al.'s VL CDR1-3, as shown in Table 1. Thus, in one embodiment, the framework region is defined based on a combination of the CDR defined by Kabat et al. and the highly variable loop defined by Chothia et al. For example, an anti-LAG-3 antibody molecule may include VH FR1 defined based on the highly variable VH loop 1 by Chothia et al., and VH FR2 defined based on VH CDR1-2 by Kabat et al., as shown in Table 1. The anti-LAG-3 antibody molecule may further include VH FR3-4 defined based on VH CDR2-3 by Kabat et al., and VL FR1-4 defined based on VL CDR1-3 by Kabat et al.

[0044] Anti-LAG-3 antibody molecules may include any combination of CDR or highly variable loops as defined by Kabat and Chothia. In one aspect, an anti-LAG-3 antibody molecule is one of the antibodies listed herein, e.g., BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum1 3, BAP050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050 (Ser) (e.g. BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050- hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050 -hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP0 The light chain variable region of an antibody selected from any of the following: 50-hum20-Ser, BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J, comprises at least 1, 2, or 3 CDRs according to the Kabat and Chothia definitions (e.g., at least 1, 2, or 3 CDRs according to the Kabat and Chothia definitions as shown in Table 1).

[0045] In some embodiments, for example, in embodiments including a variable region, a CDR (e.g., CDR or Kabat CDR), or other sequences as shown here, for example, in Table 1, the antibody molecule is a monospecific antibody molecule, a bispecific antibody molecule, or an antibody molecule including an antigen-binding fragment of an antibody, for example, a half-antibody or an antigen-binding fragment of a half-antibody. In some embodiments, the antibody molecule is a bispecific antibody molecule having first binding specificity to LAG-3 and second binding specificity to PD-1, TIM-3, CEACAM (e.g., CEACAM-1 and / or CEACAM-5), PD-L1, or PD-L2.

[0046] In one embodiment, an anti-LAG-3 antibody is (i) the VHCDR1 amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 4, or SEQ ID NO: 286; the VHCDR2 amino acid sequence of SEQ ID NO: 2; and the heavy chain variable region (VH) including the VHCDR3 amino acid sequence of SEQ ID NO: 3; and (ii) The light chain variable region (VL) containing the VLCDR1 amino acid sequence of SEQ ID NO: 10, the VLCDR2 amino acid sequence of SEQ ID NO: 11, and the VLCDR3 amino acid sequence of SEQ ID NO: 12. Includes.

[0047] In other embodiments, the anti-LAG-3 antibody molecule is (i) VHCDR1 amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 4, or SEQ ID NO: 286; heavy chain variable region (VH) including the VHCDR2 amino acid sequence of SEQ ID NO: 5 and the VHCDR3 amino acid sequence of SEQ ID NO: 3; and (ii) The light chain variable region (VL) containing the VLCDR1 amino acid sequence of SEQ ID NO: 13, the VLCDR2 amino acid sequence of SEQ ID NO: 14, and the VLCDR3 amino acid sequence of SEQ ID NO: 15. Includes.

[0048] In one embodiment, the anti-LAG-3 antibody molecule contains the VHCDR1 amino acid sequence of SEQ ID NO: 1. In another embodiment, the anti-LAG-3 antibody molecule contains the VHCDR1 amino acid sequence of SEQ ID NO: 4. In yet another embodiment, the anti-LAG-3 antibody molecule contains the VHCDR1 amino acid sequence of SEQ ID NO: 286.

[0049] In one embodiment, the light chain or heavy chain variable framework of an anti-LAG-3 antibody molecule (e.g., a region encompassing at least FR1, FR2, FR3 and optionally FR4) is: (a) a human light chain or heavy chain variable framework from a human mature antibody, human germline sequence, or human consensus sequence, e.g., a light chain or heavy chain variable framework comprising at least 80%, 85%, 87%, 90%, 92%, 93%, 95%, 97%, 98%, or preferably 100% of amino acids from light chain or heavy chain variable framework residues; (b) human mature The following can be selected: (c) a human light chain or heavy chain variable framework from an antibody, human germline sequence, or human consensus sequence, e.g., a light chain or heavy chain variable framework comprising 20% ​​to 80%, 40% to 60%, 60% to 90%, or 70% to 95% of amino acid residues from light chain or heavy chain variable framework residues; (c) a non-human framework (e.g., a rodent framework); or (d) a non-human framework that has been modified to remove antigenic or cytotoxic determinants, e.g., deimmunized or partially humanized. In one embodiment, the light chain or heavy chain variable framework region (particularly FR1, FR2 and / or FR3) comprises a light chain or heavy chain variable framework sequence that is at least 70%, 75%, 80%, 85%, 87%, 88%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99% identical to or identical to the framework of a VL or VH segment of a human germline gene.

[0050] In one embodiment, the anti-LAG-3 antibody molecule includes a heavy chain variable domain having at least 1, 2, 3, 4, 5, 6, 7, 10, 15, 20 or more changes, e.g., amino acid substitutions or deletions, from the amino acid sequence of BAP050-chi-HC, e.g., the amino acid sequence of the FR region in the entire variable region, as shown in Figures 9A-9B or SEQ ID NO: 20 or 22. In one embodiment, the anti-LAG-3 antibody molecule includes, for example, an amino acid sequence of BAP050-chi-HC, e.g., E at position 1, V at position 2, A at position 9, V at position 11, A at position 16, S at position 17, L at position 18, R at position 19, V at position 20, V or G at position 24, I at position 37, A or S at position 40, and R at position 41. Alternatively, the heavy chain variable domain may contain one or more of the following: T, S at position 42, Q or R at position 43, R at position 44, E at position 46, I or L at position 48, V at position 68, V or T at position 69, I at position 70, A at position 72, D at position 73, K at position 74, V or I at position 76, Y at position 80, W at position 83, C or S at position 84, S or T at position 85, A at position 88, E or S at position 89, V or M at position 93, or Y at position 95. In one embodiment, the antibody molecule includes a substitution in the heavy chain framework region 3 (VHFW3) (e.g., a Cys to Ser substitution at position 84) (e.g., as shown in Table 2).

[0051] In addition to or in combination with the heavy chain substitutions of BAP050-chi-HC described herein, the anti-LAG-3 antibody molecule includes a light chain variable domain having at least 1, 2, 3, 4, 5, 6, 7, 10, 15, 20 or more amino acid changes, e.g., amino acid substitutions or deletions, from the amino acid sequence of BAP050-chi-LC, e.g., the amino acid sequence shown in Figures 10A-10B or SEQ ID NO: 24 or 26. In one embodiment, the anti-LAG-3 antibody molecule has the amino acid sequence of BAP050-chi-LC, for example, the amino acid sequence shown in Figures 10A-10B or SEQ ID NO: 24 or 26, with E at position 1, V at position 3, L at position 4, S at position 7, P at position 8, A or L or D at position 9, T or F at position 10, Q at position 11, P at position 12, V or L at position 13, T at position 14, V or P at position 15, K at position 16, Q or E at position 17, T or P or K at position 18, A at position 19, and S at position 20. It includes a heavy chain variable domain having one or more of the following at position 21: L, 22: T, 37: L, 41: G, 42: K or Q, 43: A or S, 44: P, 45: R or Q, 46: L, 58: I, 60: P or D, 67: Y, 70: E, 71: F, 72: T, 73: F, 76: N, 77: S or R, 78: I, 79: Q, 80: A, S, or P, 81: D, 83: A or F, 85: Y or V, or 87: F.

[0052] In other embodiments, the anti-LAG-3 antibody molecule comprises a 1, 2, 3, or 4 heavy-chain framework region (e.g., a VHFW amino acid or nucleotide sequence encoded by the nucleotide sequences shown in Table 2 or Table 2) or a sequence substantially identical thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99%, or more identical thereto and / or has 1, 2, 3, or more substitutions, insertions, or deletions, e.g., conserved substitutions). In one embodiment, the antibody molecule comprises a substitution in heavy-chain framework region 3 (VHFW3) (e.g., a Cys-to-Ser substitution at position 84) (e.g., as shown in Table 2).

[0053] In yet another embodiment, the anti-LAG-3 antibody molecule comprises a 1, 2, 3, or 4 light chain framework region (e.g., a VLFW amino acid sequence shown in Table 2 or encoded by a nucleotide sequence shown in Table 2) or a sequence substantially identical thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical thereto and / or has 1, 2, 3 or more substitutions, insertions, or deletions, e.g., a conserved substitution).

[0054] In another embodiment, the anti-LAG-3 antibody molecule comprises a 1, 2, 3, or 4 heavy chain framework region (e.g., a VHFW amino acid sequence shown in Table 2 or encoded by a nucleotide sequence shown in Table 2) or a sequence substantially identical thereto; and a 1, 2, 3, or 4 light chain framework region (e.g., a VLFW amino acid sequence shown in Table 2 or encoded by a nucleotide sequence shown in Table 2) or a sequence substantially identical thereto.

[0055] In one respect, the anti-LAG-3 antibody molecules are BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum14, BAP050-hum15, BAP050-hum18, BAP050-hum19, BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03- Includes Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser, and heavy chain framework region 1 (VHFW1) of BAP050-Clone-F or BAP050-Clone-G (e.g., SEQ ID NO: 187). In one embodiment, the antibody molecule includes the heavy chain framework region 1 (VHFW1) of BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13 or BAP050-hum20, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-Clone-H, BAP050-Clone-I or BAP050-Clone J (e.g., SEQ ID NO: 190). In another embodiment, the antibody molecule includes the heavy chain framework region 1 (VHFW1) of BAP050-hum16 (e.g., SEQ ID NO: 194). In one embodiment, the antibody molecule contains the heavy chain framework region 1 (VHFW1) of BAP050-hum17 (e.g., SEQ ID NO: 196).In another embodiment, the antibody molecule includes a heavy chain framework region 1 (VHFW1) that has or is encoded by a sequence that is substantially identical to any of the sequences (for example, at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or has one, two, three or more substitutions, insertions or deletions, e.g., a conserved substitution).

[0056] In one respect, the anti-LAG-3 antibody molecules are BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum13, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03 -Includes heavy chain framework region 2 (VHFW2) of BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum13-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser, BAP050-Clone-F, BAP050-Clone-G, or BAP050-Clone-J (e.g., SEQ ID NO: 198). In one embodiment, the antibody molecule includes heavy chain framework region 2 (VHFW2) of BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum20, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum20-Ser, or BAP050-Clone-I (e.g., SEQ ID NO: 202). In another embodiment, the antibody molecule includes heavy chain framework region 2 (VHFW2) of BAP050-hum14, BAP050-hum15, BAP050-hum14-Ser, or BAP050-hum15-Ser (e.g., SEQ ID NO: 206). In one embodiment, the antibody molecule includes the heavy chain framework region 2 (VHFW2) of BAP050-hum16 (e.g., SEQ ID NO: 208).In another embodiment, the antibody molecule includes a heavy chain framework region 2 (VHFW2) that has or is encoded by a sequence that is substantially identical to any of the sequences (for example, at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or has one, two, three or more substitutions, insertions or deletions, e.g., a conserved substitution).

[0057] In one aspect, the anti-LAG-3 antibody molecule includes the heavy chain framework region 3 (VHFW3) of BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BAP050-hum15, BAP050-hum18, BAP050-hum19, or BAP050-hum20 (e.g., SEQ ID NO: 210). In one respect, the antibody molecules are BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Se r, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser, BAP050-hum20-Ser, BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J include heavy chain framework region 3 (VHFW3) (e.g., SEQ ID NO: 212). In one embodiment, the antibody molecule includes heavy chain framework region 3 (VHFW3) of BAP050-hum16 (e.g., SEQ ID NO: 217). In one embodiment, the antibody molecule includes heavy chain framework region 3 (VHFW3) of BAP050-hum17 (e.g., SEQ ID NO: 219).In another embodiment, the antibody molecule includes a heavy chain framework region 3 (VHFW3) that has or is encoded by a sequence that is substantially identical to any of the sequences (for example, at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or has one, two, three or more substitutions, insertions or deletions, e.g., a conserved substitution).

[0058] In one respect, the anti-LAG-3 antibody molecules are BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum1 2, BAP050-hum13, BAP050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-h um19 or BAP050-hum20, BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BA P050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP05 In other embodiments, the antibody molecule includes a heavy chain framework region 4 (VHFW4) of the following sequence: 0-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser, BAP050-hum20-Ser, BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J (e.g., SEQ ID NO: 221). In other embodiments, the antibody molecule includes a heavy chain framework region 4 (VHFW4) having or encoded by a sequence that is substantially identical to any of the aforementioned sequences (e.g., at least about 85%, 90%, 95%, 99% or more identical to any of the aforementioned sequences and / or having one, two, three or more substitutions, insertions, or deletions, e.g., conserved substitutions).

[0059] In one respect, anti-LAG-3 antibody molecules include BAP050-hum01, BAP050-hum02, BAP050-hum04, BAP050-hum07, BAP050-hum09, BAP050-hum11, BAP050-hum13, BAP050-hum17, BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum04-Ser, and BA. The antibody molecule contains light chain framework region 1 (VLFW1) of P050-hum07-Ser, BAP050-hum09-Ser, BAP050-hum11-Ser, BAP050-hum13-Ser, BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J (e.g., SEQ ID NO: 226). In one embodiment, the antibody molecule contains light chain framework region 1 (VLFW1) of BAP050-hum03, BAP050-hum10, BAP050-hum14, BAP050-hum03-Ser, BAP050-hum10-Ser, or BAP050-hum14-Ser (e.g., SEQ ID NO: 230). In one embodiment, the antibody molecule includes a light chain framework region 1 (VLFW1) of BAP050-hum05 or BAP050-hum05-Ser (e.g., SEQ ID NO: 232). In another embodiment, the antibody molecule includes a light chain framework region 1 (VLFW1) of BAP050-hum06, BAP050-hum20, BAP050-hum06-Ser or BAP050-hum20-Ser (e.g., SEQ ID NO: 234). In one embodiment, the antibody molecule includes a light chain framework region 1 (VLFW1) of BAP050-hum08, BAP050-hum12, BAP050-hum15, BAP050-hum16, BAP050-hum19, BAP050-hum08-Ser, BAP050-hum12-Ser, BAP050-hum15-Ser, or BAP050-hum19-Ser (e.g., SEQ ID NO: 236). In another embodiment, the antibody molecule includes a light chain framework region 1 (VLFW1) of BAP050-hum18 or BAP050-hum18-Ser (e.g., SEQ ID NO: 238).In another embodiment, the antibody molecule includes a light chain framework region 1 (VLFW1) that has or is encoded by a sequence that is substantially identical to any of the sequences (for example, at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or has one, two, three or more substitutions, insertions or deletions, e.g., a conserved substitution).

[0060] In one aspect, the anti-LAG-3 antibody molecule includes the light chain framework region 2 (VLFW2) of BAP050-hum01, BAP050-hum02, BAP050-hum05, BAP050-hum09, BAP050-hum13, BAP050-hum17, BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum05-Ser, BAP050-hum09-Ser, BAP050-hum13-Ser, BAP050-hum17-Ser, BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J (e.g., SEQ ID NO: 240). In one respect, the antibody molecules are BAP050-hum03, BAP050-hum06, BAP050-hum08, BAP050-hum10, BAP050-hum12, BAP050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum18, BAP050-hum19, BAP050-hum20, BAP050-hum03-Ser, BAP050-h The antibody molecule contains a light chain framework region 2 (VLFW2) of um06-Ser, BAP050-hum08-Ser, BAP050-hum10-Ser, BAP050-hum12-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser, or BAP050-hum20-Ser (e.g., SEQ ID NO: 244). In one embodiment, the antibody molecule contains a light chain framework region 2 (VLFW2) of BAP050-hum04 or BAP050-hum04-Ser (e.g., SEQ ID NO: 246). In one embodiment, the antibody molecule includes a light chain framework region 2 (VLFW2) of BAP050-hum07, BAP050-hum11, BAP050-hum07-Ser, or BAP050-hum11-Ser (e.g., SEQ ID NO: 248).In another embodiment, the antibody molecule includes a light chain framework region 2 (VLFW2) that has or is encoded by a sequence that is substantially identical to any of the sequences (for example, at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or has one, two, three or more substitutions, insertions or deletions, e.g., a conserved substitution).

[0061] In one aspect, the anti-LAG-3 antibody molecule includes the light chain framework region 3 (VLFW3) of BAP050-hum01, BAP050-hum03, BAP050-hum05, BAP050-hum10, BAP050-hum14, BAP050-hum19, BAP050-hum01-Ser, BAP050-hum03-Ser, BAP050-hum05-Ser, BAP050-hum10-Ser, BAP050-hum14-Ser, BAP050-hum19-Ser, or BAP050-Clone-F (e.g., SEQ ID NO: 252). In one embodiment, the antibody molecule includes a light chain framework region 3 (VLFW3) of BAP050-hum02, BAP050-hum09, BAP050-hum13, BAP050-hum02-Ser, BAP050-hum09-Ser, BAP050-hum13-Ser, BAP050-Clone-G, BAP050-Clone-H, or BAP050-Clone-J (e.g., SEQ ID NO: 255). In another embodiment, the antibody molecule includes a light chain framework region 3 (VLFW3) of BAP050-hum04 or BAP050-hum04-Ser (e.g., SEQ ID NO: 259). In one embodiment, the antibody molecule includes a light chain framework region 3 (VLFW3) of BAP050-hum06, BAP050-hum07, BAP050-hum11, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum11-Ser, or BAP050-Clone-I (e.g., SEQ ID NO: 261). In another embodiment, the antibody molecule includes a light chain framework region 3 (VLFW3) of BAP050-hum08, BAP050-hum12, BAP050-hum15, BAP050-hum16, BAP050-hum18, BAP050-hum08-Ser, BAP050-hum12-Ser, BAP050-hum15-Ser, or BAP050-hum18-Ser (e.g., SEQ ID NO: 265). In one embodiment, the antibody molecule includes the light chain framework region 3 (VLFW3) of BAP050-hum17 (e.g., SEQ ID NO: 267).In one embodiment, the antibody molecule includes a light chain framework region 3 (VLFW3) of BAP050-hum20 or BAP050-hum20-Ser (e.g., SEQ ID NO: 269). In another embodiment, the antibody molecule includes a light chain framework region 3 (VHLW3) having or encoded by a sequence that is substantially identical to any of the aforementioned sequences (e.g., at least about 85%, 90%, 95%, 99% or more identical to any of the aforementioned sequences and / or having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0062] In one respect, the anti-LAG-3 antibody molecules are BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum1 2, BAP050-hum13, BAP050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050- hum19, BAP050-hum20, BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, B AP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP 050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050 In other embodiments, the antibody molecule includes a light chain framework region 4 (VLFW4) of -hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser, BAP050-hum20-Ser, BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J (e.g., SEQ ID NO: 271). In other embodiments, the antibody molecule includes a light chain framework region 4 (VLFW4) having or encoded by a sequence that is substantially identical to any of the sequences described above (e.g., at least about 85%, 90%, 95%, 99% or more identical to any of the sequences described above and / or having one, two, three or more substitutions, insertions, or deletions, e.g., conserved substitutions).

[0063] In one embodiment, the anti-LAG-3 antibody molecule includes heavy chain framework regions 1-3 of BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum18, and BAP050-hum19 (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 210 (VHFW3)). In one embodiment, the antibody molecule includes heavy chain framework regions 1-3 of BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, and BAP050-hum20 (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 202 (VHFW2), and SEQ ID NO: 210 (VHFW3)). In one embodiment, the antibody molecule includes heavy chain framework regions 1-3 of BAP050-hum13 (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 210 (VHFW3)). In one embodiment, the antibody molecule includes heavy chain framework regions 1-3 of BAP050-hum14 or BAP050-hum15 (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 206 (VHFW2), and SEQ ID NO: 210 (VHFW3)). In one embodiment, the antibody molecule includes heavy chain framework regions 1-3 of BAP050-hum16 (e.g., SEQ ID NO: 194 (VHFW1), SEQ ID NO: 208 (VHFW2), and SEQ ID NO: 217 (VHFW3)). In another embodiment, the antibody molecule includes heavy chain framework regions 1-3 of BAP050-hum17 (e.g., SEQ ID NO: 196 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 219 (VHFW3)).In one embodiment, the antibody molecule includes heavy chain framework regions 1-3 of BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser, BAP050-Clone-F, or BAP050-Clone-G (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 212 (VHFW3)). In one embodiment, the antibody molecule includes heavy chain framework regions 1-3 of BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum20-Ser, BAP050-Clone-H, or BAP050-Clone-I (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 202 (VHFW2), and SEQ ID NO: 212 (VHFW3)). In another embodiment, the antibody molecule includes heavy chain framework regions 1-3 of BAP050-hum13-Ser or BAP050-Clone-J (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 212 (VHFW3)). In one embodiment, the antibody molecule includes heavy chain framework regions 1-3 of BAP050-hum14-Ser or BAP050-hum15-Ser (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 206 (VHFW2), and SEQ ID NO: 212 (VHFW3)).In one respect, the antibody molecule is further divided into BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP0 Includes heavy chain framework region 4 of 50-hum18, BAP050-hum19, BAP050-hum20, BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum05-Ser, BAP050-hum09-Ser, BAP050-hum11-Ser, BAP050-hum13-Ser, BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J (e.g., SEQ ID NO: 221). In another embodiment, the antibody molecule includes a heavy chain framework region having or encoded by a sequence that is substantially identical to any of the sequences (for example, at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or having one, two, three or more substitutions, insertions or deletions, e.g., a conserved substitution).

[0064] In one embodiment, the anti-LAG-3 antibody molecule includes light chain framework regions 1-3 of BAP050-hum01, BAP050-hum01-Ser, or BAP050-Clone-F (e.g., SEQ ID NO: 226 (VLFW1), SEQ ID NO: 240 (VLFW2), and SEQ ID NO: 252 (VLFW3)). In another embodiment, the antibody molecule includes light chain framework regions 1-3 of BAP050-hum02, BAP050-hum09, BAP050-hum13, BAP050-hum02-Ser, BAP050-hum09-Ser, BAP050-hum13-Ser, BAP050-Clone-G, BAP050-Clone-H, or BAP050-Clone-J (e.g., SEQ ID NO: 226 (VLFW1), SEQ ID NO: 240 (VLFW2), and SEQ ID NO: 255 (VLFW3)). In one embodiment, the antibody molecule includes light chain framework regions 1-3 of BAP050-hum03, BAP050-hum10, BAP050-hum14, BAP050-hum03-Ser, BAP050-hum10-Ser, or BAP050-hum14-Ser (e.g., SEQ ID NO: 230 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 252 (VLFW3)). In another embodiment, the antibody molecule includes light chain framework regions 1-3 of BAP050-hum04 or BAP050-hum04-Ser (e.g., SEQ ID NO: 226 (VLFW1), SEQ ID NO: 246 (VLFW2), and SEQ ID NO: 259 (VLFW3)). In one embodiment, the antibody molecule includes light chain framework regions 1-3 of BAP050-hum05 or BAP050-hum05-Ser (e.g., SEQ ID NO: 232 (VLFW1), SEQ ID NO: 240 (VLFW2), and SEQ ID NO: 252 (VLFW3)). In another embodiment, the antibody molecule includes light chain framework regions 1-3 of BAP050-hum06 or BAP050-hum06-Ser (e.g., SEQ ID NO: 234 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 261 (VLFW3)).In one embodiment, the antibody molecule includes light chain framework regions 1-3 of BAP050-hum07, BAP050-hum11, BAP050-hum07-Ser, BAP050-hum11-Ser, or BAP050-Clone-I (e.g., SEQ ID NO: 226 (VLFW1), SEQ ID NO: 248 (VLFW2), and SEQ ID NO: 261 (VLFW3)). In another embodiment, the antibody molecule includes light chain framework regions 1-3 of BAP050-hum08, BAP050-hum12, BAP050-hum15, BAP050-hum16, BAP050-hum08-Ser, BAP050-hum12-Ser, or BAP050-hum15-Ser (e.g., SEQ ID NO: 236 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 265 (VLFW3)). In one embodiment, the antibody molecule includes light chain framework regions 1-3 of BAP050-hum17 (e.g., SEQ ID NO: 226 (VLFW1), SEQ ID NO: 240 (VLFW2), and SEQ ID NO: 267 (VLFW3)). In one embodiment, the antibody molecule includes light chain framework regions 1-3 of BAP050-hum18 or BAP050-hum18-Ser (e.g., SEQ ID NO: 238 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 265 (VLFW3)). In one embodiment, the antibody molecule includes light chain framework regions 1-3 of BAP050-hum19 or BAP050-hum19-Ser (e.g., SEQ ID NO: 236 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 252 (VLFW3)). In one embodiment, the antibody molecule comprises light chain framework regions 1-3 of BAP050-hum20 or BAP050-hum20-Ser (e.g., SEQ ID NO: 234 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 269 (VLFW3)).In one respect, the antibody molecule is further divided into BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum1 2, BAP050-hum13, BAP050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050 -hum19, BAP050-hum20, BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser , BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, B In other embodiments, the antibody molecule includes a heavy chain framework region 4 of AP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser, BAP050-hum20-Ser, BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J (e.g., SEQ ID NO: 271). In other embodiments, the antibody molecule includes a light chain framework region having or encoded by a sequence that is substantially identical to any of the sequences described above (e.g., at least about 85%, 90%, 95%, 99% or more identical to any of the sequences described above and / or having one, two, three or more substitutions, insertions, or deletions, e.g., conserved substitutions).

[0065] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum01 (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or heavy chain framework regions 1-3 of BAP050-hum01-Ser or BAP050-Clone-F (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum01, BAP050-hum01-Ser, or BAP050-Clone-F (e.g., SEQ ID NO: 226 (VLFW1), SEQ ID NO: 240 (VLFW2), and SEQ ID NO: 252 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0066] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum02 (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or heavy chain framework regions 1-3 of BAP050-hum02-Ser or BAP050-Clone-G (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum02, BAP050-hum02-Ser, or BAP050-Clone-G (e.g., SEQ ID NO: 226 (VLFW1), SEQ ID NO: 240 (VLFW2), and SEQ ID NO: 255 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0067] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum03 (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or heavy chain framework regions 1-3 of BAP050-hum03-Ser (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum03 (e.g., SEQ ID NO: 230 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 252 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0068] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum04 (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or heavy chain framework regions 1-3 of BAP050-hum04-Ser (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum04 (e.g., SEQ ID NO: 226 (VLFW1), SEQ ID NO: 246 (VLFW2), and SEQ ID NO: 259 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0069] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum05 or BAP050-hum05-Ser (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 210 (VHFW3)) (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum05 or BAP050-hum05-Ser (e.g., SEQ ID NO: 232 (VLFW1), SEQ ID NO: 240 (VLFW2), and SEQ ID NO: 252 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0070] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum06 (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or heavy chain framework regions 1-3 of BAP050-hum06-Ser (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum06 (e.g., SEQ ID NO: 234 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 261 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0071] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum07 (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or heavy chain framework regions 1-3 of BAP050-hum07-Ser (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum07 (e.g., SEQ ID NO: 226 (VLFW1), SEQ ID NO: 248 (VLFW2), and SEQ ID NO: 261 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0072] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum08 (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or heavy chain framework regions 1-3 of BAP050-hum08-Ser (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum08 (e.g., SEQ ID NO: 236 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 265 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0073] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum09 (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 202 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or BAP050-hum09-Ser or BAP050-Clone-H (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 202 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum09, BAP050-hum09-Ser, or BAP050-Clone-H (e.g., SEQ ID NO: 226 (VLFW1), SEQ ID NO: 240 (VLFW2), and SEQ ID NO: 255 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0074] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum10 (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 202 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or heavy chain framework regions 1-3 of BAP050-hum10-Ser (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 202 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum10 (e.g., SEQ ID NO: 230 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 252 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0075] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum11 (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 202 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or BAP050-hum11-Ser or BAP050-Clone-I (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 202 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum11, BAP050-hum11-Ser, or BAP050-Clone-I (e.g., SEQ ID NO: 226 (VLFW1), SEQ ID NO: 248 (VLFW2), and SEQ ID NO: 261 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0076] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum12 (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 202 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or BAP050-hum12-Ser (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 202 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum12 or BAP050-hum12-Ser (e.g., SEQ ID NO: 236 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 265 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0077] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum13 (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or heavy chain framework regions 1-3 of BAP050-hum13-Ser or BAP050-Clone-J (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum13, BAP050-hum13-Ser, or BAP050-Clone-J (e.g., SEQ ID NO: 226 (VLFW1), SEQ ID NO: 240 (VLFW2), and SEQ ID NO: 255 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0078] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum14 (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 206 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or heavy chain framework regions 1-3 of BAP050-hum14-Ser (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 206 (VHFW2), and SEQ ID NO: 210 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum14 (e.g., SEQ ID NO: 230 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 252 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0079] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum15 (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 206 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or heavy chain framework regions 1-3 of BAP050-hum15-Ser (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 206 (VHFW2), and SEQ ID NO: 210 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum15 (e.g., SEQ ID NO: 236 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 265 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0080] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum16 (e.g., SEQ ID NO: 194 (VHFW1), SEQ ID NO: 208 (VHFW2), and SEQ ID NO: 217 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum16 (e.g., SEQ ID NO: 236 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 265 (VLFW3)). In another embodiment, the antibody molecule comprises heavy chain and light chain framework regions having or encoding a sequence that is substantially identical to any of the sequences (e.g., at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or having one, two, three or more substitutions, insertions, or deletions, e.g., conserved substitutions).

[0081] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum17 (e.g., SEQ ID NO: 196 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 219 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum17 (e.g., SEQ ID NO: 226 (VLFW1), SEQ ID NO: 240 (VLFW2), and SEQ ID NO: 267 (VLFW3)). In another embodiment, the antibody molecule comprises heavy chain and light chain framework regions having or encoding a sequence that is substantially identical to any of the sequences (e.g., at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or having one, two, three or more substitutions, insertions, or deletions, e.g., conserved substitutions).

[0082] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum18 (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or heavy chain framework regions 1-3 of BAP050-hum18-Ser (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum18 (e.g., SEQ ID NO: 238 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 265 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0083] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum19 (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or heavy chain framework regions 1-3 of BAP050-hum18-Ser (e.g., SEQ ID NO: 187 (VHFW1), SEQ ID NO: 198 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum19 (e.g., SEQ ID NO: 236 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 252 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0084] In one embodiment, the anti-LAG-3 antibody molecule comprises heavy chain framework regions 1-3 of BAP050-hum20 (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 202 (VHFW2), and SEQ ID NO: 210 (VHFW3)) or BAP050-hum20-Ser (e.g., SEQ ID NO: 190 (VHFW1), SEQ ID NO: 202 (VHFW2), and SEQ ID NO: 212 (VHFW3)); and light chain framework regions 1-3 of BAP050-hum20 (e.g., SEQ ID NO: 234 (VLFW1), SEQ ID NO: 244 (VLFW2), and SEQ ID NO: 269 (VLFW3)). In another embodiment, the antibody molecule includes heavy and light chain framework regions having or encoded by sequences that are substantially identical to any of the sequences (for example, sequences that are at least about 85%, 90%, 95%, 99% or more identical to any of the sequences and / or sequences having one, two, three or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0085] In one embodiment, the anti-LAG-3 antibody molecule includes a heavy chain framework region having a combination of framework regions FW1, FW2, and FW3, as shown in Figure 4 or 6. In another embodiment, the antibody molecule includes a light chain framework region having a combination of framework regions FW1, FW2, and FW3, as shown in Figure 4 or 6. In yet another embodiment, the antibody molecule includes a heavy chain framework region having a combination of framework regions FW1, FW2, and FW3, as shown in Figure 4 or 6, and a light chain framework region having a combination of framework regions FW1, FW2, and FW3, as shown in Figure 4 or 6.

[0086] In one aspect, the heavy chain or light chain variable domain or both of the anti-LAG-3 antibody molecule has an amino acid sequence substantially identical to the amino acids disclosed herein, for example, the antibodies described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum07 m08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BAP050-hum1 5, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g., BAP050-h um01-Ser, BAP050-hum02-Ser, BAP050-hum05-Ser, BAP050-hum09-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser , BAP050-hum13-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I or BAP050-Clone -An antibody selected from any of the following, which matches the variable region of the antibody by at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, or more; or as described in Table 1 or encoded by the nucleotide sequence in Table 1; or including at least 1 or 5 residues, but less than 40, 30, 20, or 10 residues, from the variable region of the antibodies described herein.

[0087] In one embodiment, the heavy chain or light chain variable region or both of the anti-LAG-3 antibody molecule comprises the nucleic acid sequence described herein or an amino acid sequence encoded by a nucleic acid (e.g., a specific nucleic acid sequence or nucleic acid sequence encoding the amino acid sequence described herein, such as those shown in Tables 1 and 2) or its complement, which hybridizes with the nucleic acid sequence described herein under, for example, low stringency, medium stringency, high stringency, or other hybridization conditions described herein.

[0088] In another embodiment, the anti-LAG-3 antibody molecule includes at least 1, 2, 3, or 4 antigen-binding regions, e.g., variable regions, having the amino acid sequences shown in Table 1 or sequences substantially identical thereto (e.g., sequences identical by at least about 85%, 90%, 95%, 99%, or more, or sequences that do not differ by more than 1, 2, 5, 10, or 15 amino acid residues from the sequences shown in Table 1). In another embodiment, the anti-LAG-3 antibody molecule includes VH and / or VL domains encoded by nucleic acids having the nucleotide sequences shown in Table 1 or sequences substantially identical thereto (e.g., sequences identical by at least about 70%, 75%, 85%, 90%, 95%, 99%, or more, or sequences that do not differ by more than 3, 6, 15, 30, or 45 nucleotides from the sequences shown in Table 1).

[0089] In yet another embodiment, the anti-LAG-3 antibody molecule comprises at least 1, 2, or 3 CDRs from a heavy chain variable region having the amino acid sequence shown in Table 1 or a sequence substantially homologous thereto (e.g., a sequence identical to it by at least about 85%, 90%, 95%, 99%, or more, and / or a sequence having 1, 2, 3, or more substitutions, insertions, or deletions, e.g., conserved substitutions). In yet another embodiment, the anti-LAG-3 antibody molecule comprises at least 1, 2, or 3 CDRs from a light chain variable region having the amino acid sequence shown in Table 1 or a sequence substantially homologous thereto (e.g., a sequence identical to it by at least about 85%, 90%, 95%, 99%, or more, and / or a sequence having 1, 2, 3, or more substitutions, insertions, or deletions, e.g., conserved substitutions). In yet another embodiment, the anti-LAG-3 antibody molecule comprises at least 1, 2, 3, 4, 5, or 6 CDRs from the heavy and light chain variable regions having the amino acid sequences shown in Table 1 or sequences substantially homologous thereto (e.g., sequences that are at least about 85%, 90%, 95%, 99% or more identical thereto and / or sequences having 1, 2, 3 or more substitutions, insertions or deletions, e.g., conserved substitutions). In one embodiment, at least 1, 2, 3, 4, 5, or 6 CDRs are defined by Kabat, for example, as shown in Table 1. In another embodiment, at least 1, 2, 3, 4, 5, or 6 CDRs are defined by Chothia, for example, as shown in Table 1.

[0090] In one aspect, the anti-LAG-3 antibody molecule is the antibody described herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum08, BAP050-hum09, BAP050-hum09, as summarized in Table 1. m10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BAP050-hum15, BAP050-hum16, BAP0 50-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g., BAP050-hum01-Ser, BAP0 The antibody comprises at least 1, 2, or 3 CDRs and / or highly variable loops from a heavy chain variable region having an amino acid sequence of an antibody selected from any of the following: 50-hum02-Ser, BAP050-hum05-Ser, BAP050-hum09-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J, or a sequence substantially identical thereto (e.g., at least about 85%, 90%, 95%, 99% or more identical thereto and / or a sequence having 1, 2, 3 or more substitutions, insertions, or deletions, e.g., a conserved substitution).In other aspects, anti-LAG-3 antibody molecules include the antibodies listed herein, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050 -hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BAP050-hum15, BAP050-hum16 , BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g., BAP050-hum01-S The antibody comprises at least 1, 2, or 3 CDRs from a light chain variable region having the amino acid sequence of an antibody selected from any of the following: er, BAP050-hum02-Ser, BAP050-hum05-Ser, BAP050-hum09-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J, or a sequence substantially identical thereto (e.g., at least about 85%, 90%, 95%, 99% or more identical thereto and / or having 1, 2, 3 or more substitutions, insertions, or deletions, e.g., a sequence having conserved substitutions). In one embodiment, the anti-LAG-3 antibody molecule comprises a total of 6 CDRs and / or highly variable loops, as described herein, for example, as shown in Table 1.

[0091] In one embodiment, the anti-LAG-3 antibody molecule includes a variable region that is identical in sequence to the variable region described herein (e.g., the FR region disclosed herein) or differs by one, two, three, or four amino acids.

[0092] In one embodiment, the anti-LAG-3 antibody molecule is a complete antibody or a fragment thereof (e.g., Fab, F(ab')2, Fv, or a single-chain Fv fragment (scFv)). In one embodiment, the anti-LAG-3 antibody molecule is a monoclonal antibody or a monospecific antibody. The anti-LAG-3 antibody molecule may also be a humanized, chimeric, camel, shark, or in vitro-generated antibody molecule. In one embodiment, the anti-LAG-3 antibody molecule is a humanized antibody molecule. The heavy and light chains of the anti-LAG-3 antibody molecule may be full-length (e.g., the antibody may contain at least one, preferably two, complete heavy chains and at least one, preferably two, complete light chains) or antigen-binding fragments (e.g., Fab, F(ab')2, Fv, a single-chain Fv fragment, a single-domain antibody, a bispecific antibody (dAb), a bivalent antibody or a bispecific antibody or a fragment thereof, its single-domain variant, or a camel antibody).

[0093] In one embodiment, the anti-LAG-3 antibody molecule is in the form of a bispecific or polyspecific antibody molecule. In one embodiment, the bispecific antibody molecule has a first binding specificity to LAG-3 and a second binding specificity to PD-1, TIM-3, CEACAM (e.g., CEACAM-1 and / or CEACAM-5), PD-L1, or PD-L2. In one embodiment, the bispecific antibody molecule binds to LAG-3 and PD-1. In another embodiment, the bispecific antibody molecule binds to LAG-3 and TIM-3. In yet another embodiment, the bispecific antibody molecule binds to LAG-3 and CEACAM (e.g., CEACAM-1 and / or CEACAM-5). In yet another embodiment, the bispecific antibody molecule binds to LAG-3 and CEACAM-1. In yet another embodiment, the bispecific antibody molecule binds to LAG-3 and CEACAM-5. In yet another embodiment, the bispecific antibody molecule binds to LAG-3 and PD-L1. In yet another embodiment, the bispecific antibody molecule binds to LAG-3 and PD-L2. Any combination of the aforementioned molecules can be used to form a multispecific antibody molecule, for example, a triplicate antibody having primary binding specificity to LAG-3 and secondary and tertiary binding specificities to one or more of PD-1, TIM-3, CEACAM (e.g., CEACAM-1 or CEACAM-5), PD-L1, or PD-L2.

[0094] In other embodiments, an anti-LAG-3 antibody molecule is used in combination with a bispecific molecule containing one or more of PD-1, TIM-3, CEACAM (e.g., CEACAM-1 or CEACAM-5), PD-L1, or PD-L2. In one embodiment, the bispecific antibody molecule used in the combination binds to CEACAM (e.g., CEACAM-1 and / or CEACAM-5) and PD-1. In another embodiment, the bispecific antibody molecule used in the combination binds to CEACAM (e.g., CEACAM-1 and / or CEACAM-5) and TIM-3. In yet another embodiment, the bispecific antibody molecule used in the combination binds to PD-1 and TIM-3.

[0095] In yet another embodiment, the anti-LAG-3 antibody molecule is selected from, for example, the heavy chain constant regions of IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE; in particular, it has a heavy chain constant region (Fc) of, for example, the heavy chain constant region of IgG1, IgG2, IgG3, and IgG4, more specifically, the heavy chain constant region of IgG1, IgG2, or IgG4 (e.g., human IgG1, IgG2, or IgG4). In one embodiment, the heavy chain constant region is human IgG1 or human IgG4. In another embodiment, the anti-LAG-3 antibody molecule has a light chain constant region selected from, for example, the light chain constant region of kappa or lambda, preferably kappa (e.g., human kappa). In one embodiment, the constant region is modified, e.g., mutated, for example, to modify the properties of the anti-LAG-3 antibody molecule (e.g., for an increase or decrease of one or more Fc receptor binding, antibody glycosylation, cysteine ​​residue count, effector cell function, or complement function). For example, the constant region is mutated at positions 296 (M to Y), 298 (S to T), 300 (T to E), 477 (H to K), and 478 (N to F) to alter Fc receptor binding (for example, the mutation sites correspond to positions 132 (M to Y), 134 (S to T), 136 (T to E), 313 (H to K), and 314 (N to F) in sequence numbers 212 or 214; or positions 135 (M to Y), 137 (S to T), 139 (T to E), 316 (H to K), and 317 (N to F) in sequence numbers 215, 216, 217, or 218). In another embodiment, the heavy chain constant region of IgG4, for example, human IgG4, is mutated at position 228 according to EU numbering (e.g., S to P), as shown in Table 3. In one embodiment, the anti-LAG-3 antibody molecule includes, for example, human IgG4 with a mutation at position 228 of EU numbering (e.g., S to P), as shown in Table 3, and, for example, the kappa light chain constant region, as shown in Table 3.In yet another embodiment, the heavy chain constant region of IgG1, for example, human IgG1, is mutated at one or more EU numbering positions, such as position 297 (e.g., N to A), position 265 (e.g., D to A), position 329 (e.g., P to A), position 234 (e.g., L to A), or position 235 (e.g., L to A), as shown in Table 3. In one embodiment, the anti-LAG-3 antibody molecule includes, for example, human IgG1 with one or more of the aforementioned positions mutated, as shown in Table 3; and, for example, the kappa light chain constant region shown in Table 3.

[0096] In one embodiment, the anti-LAG-3 antibody molecule is either isolated or recombinant. In one embodiment, it is a humanized antibody molecule that is an anti-LAG-3 antibody molecule.

[0097] In one embodiment, anti-LAG-3 antibody molecules have risk scores of 1200, 1150, 1100, 1050, 1000, 950, 900, 850, or less than 800, based on T cell epitope analysis.

[0098] In one embodiment, the anti-LAG-3 antibody molecule is a humanized antibody molecule having a risk score of 800-1200, 850-1150, 900-1100, 950-1050, or any of the risk scores listed herein, based on T cell epitope analysis.

[0099] The present invention also relates to nucleic acid molecules comprising one or more nucleotide sequences encoding heavy and light chain variable regions, CDRs, highly variable loops and / or framework regions of an anti-LAG-3 antibody molecule, as described herein. In some embodiments, the nucleotide sequences encoding the anti-LAG-3 antibody molecule are codon-optimized. For example, the present invention relates to, for example, BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum 13, BAP050-hum14, BAP050-hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, h uBAP050(Ser) (e.g. BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050-hum04-Ser, BAP050-hum0 5-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP0 50-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Se This relates to the first and second nucleic acids, or substantially identical sequences thereof, encoding the heavy chain and light chain variable regions, of an anti-LAG-3 antibody molecule selected from one or more of the following: r, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I or BAP050-Clone-J, respectively.For example, nucleic acids may include nucleotide sequences shown in Tables 1 and 2 or sequences that are substantially identical thereto (e.g., sequences that are at least about 85%, 90%, 95%, 99% or more identical thereto, or sequences that do not differ by more than 3, 6, 15, 30, or 45 nucleotides from the sequences shown in Tables 1 and 2).

[0100] In other embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding a heavy chain variable domain and a heavy chain constant region, including the amino acid sequences of BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J; or one encoded by the nucleotide sequences listed in Table 1 or in Table 1; or a sequence substantially identical to any of the aforementioned sequences (e.g., matching at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, or higher).

[0101] In other embodiments, the nucleic acid comprises a nucleotide sequence encoding a light chain variable domain and / or light chain constant region, including the amino acid sequence of BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone-I, or BAP050-Clone-J; or one encoded by the nucleotide sequences listed in Table 1 or in Table 1; or a sequence substantially identical to any of the aforementioned sequences (e.g., matching at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99%, or higher).

[0102] The nucleotide sequences encoding the anti-LAG-3 heavy and light chain variable domains and constant region can be present in separate nucleic acid molecules or in the same nucleic acid molecule. In one embodiment, the nucleic acid molecule includes a nucleotide sequence encoding a leader sequence, for example, the leader sequences shown in Table 4 or a sequence substantially identical thereto.

[0103] In one embodiment, the nucleic acid molecule comprises a nucleotide sequence encoding at least 1, 2, or 3 CDRs or highly variable loops from a heavy chain variable region, having the amino acid sequences shown in Table 1 or sequences substantially homologous thereto (e.g., sequences that are at least about 85%, 90%, 95%, 99% or more identical thereto and / or sequences having 1, 2, 3 or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0104] In other embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding at least 1, 2, or 3 CDRs or highly variable loops from a light chain variable region having the amino acid sequences shown in Table 1 or sequences substantially homologous thereto (e.g., sequences that are at least about 85%, 90%, 95%, 99% or more identical thereto and / or sequences having 1, 2, 3 or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0105] In yet another embodiment, the nucleic acid molecule may include nucleotide sequences encoding at least 1, 2, 3, 4, 5, or 6 CDRs or highly variable loops from heavy and light chain variable regions having the amino acid sequences shown in Table 1 or sequences substantially homologous thereto (e.g., sequences that are at least about 85%, 90%, 95%, 99% or more identical thereto and / or sequences having 1, 2, 3 or more substitutions, insertions or deletions, e.g., conserved substitutions).

[0106] In one embodiment, the nucleic acid molecule comprises a nucleotide sequence encoding an anti-LAG-3 antibody molecule, which includes a substitution in the heavy chain framework region 3 (VHFW3) (e.g., a Cys-to-Ser substitution at position 84) (e.g., as shown in Tables 1 and 2).

[0107] In other aspects, nucleic acid molecules are summarized in Tables 1 and 2 as follows: BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BAP050-hum15, BAP050-hum 16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g. BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum 03-Ser, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hu One or more heavy chains from among m10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP049-Clone-F, BAP049-Clone-G, BAP049-Clone-H, BAP049-Clone-I or BAP049-Clone-J The framework region includes, for example, VHFW1(type a), VHFW1(type b), VHFW1(type c), VHFW1(type d), VHFW2(type a), VHFW2(type b), VHFW2(type c), VHFW2(type d), VHFW3(type a), VHFW3(type a'), VHFW3(type b), VHFW3(type c), or VHFW4 or any combination thereof, for example, any of the framework combinations listed herein) or an array that is substantially identical thereto.For example, nucleic acid molecules may contain nucleotide sequences shown in Tables 1 and 2 or sequences substantially identical thereto (e.g., sequences that are at least about 85%, 90%, 95%, 99% or more identical thereto, or sequences that do not differ by more than 3, 6, 15, 30, or 45 nucleotides from the sequences shown in Tables 1 and 2).

[0108] In other aspects, nucleic acid molecules are summarized in Tables 1 and 2 as follows: BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05, BAP050-hum06, BAP050-hum07, BAP050-hum08, BAP050-hum09, BAP050-hum10, BAP050-hum11, BAP050-hum12, BAP050-hum13, BAP050-hum14, BAP050-hum15, BAP050-hum16 , BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (e.g., BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-S er, BAP050-hum04-Ser, BAP050-hum05-Ser, BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-S One or more light chain frameworks from among BAP050-hum11-Ser, BAP050-hum12-Ser, BAP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP049-Clone-F, BAP049-Clone-G, BAP049-Clone-H, BAP049-Clone-I or BAP049-Clone-J Domains (for example, VLFW1 (type a), VLFW1 (type b), VLFW1 (type c), VLFW1 (type d), VLFW1 (type e), VLFW1 (type f), VLFW2 (type a), VLFW2 (type b), VLFW2 (type c), VLFW2 (type d), VLFW3 (type a), VLFW3 (type b), VLFW3 (type c), VLFW3 (type d), VLFW3 (type e), VLFW3 (type f), VLFW3 (type g), or VLFW4 or any combination thereof, for example,Nucleic acid molecules may contain any of the framework combinations described herein or sequences that are substantially identical thereto. For example, nucleic acid molecules may contain nucleotide sequences shown in Tables 1 and 2 or sequences that are substantially identical thereto (e.g., sequences that are at least about 85%, 90%, 95%, 99% or more identical thereto, or sequences that do not differ by more than 3, 6, 15, 30, or 45 nucleotides from the sequences shown in Tables 1 and 2).

[0109] In other embodiments, the nucleic acid molecule comprises one or more heavy chain framework regions and one or more light chain framework regions as described herein. The heavy chain and light chain framework regions may reside in the same vector or in different vectors.

[0110] In other words, this application relates to host cells and vectors containing the nucleic acids described herein. The nucleic acids may be present in a single vector or in different vectors present in the same host cell or different host cells. The host cells may be eukaryotic cells, e.g., mammalian cells, insect cells, yeast cells, or prokaryotic cells, e.g., Escherichia coli. Mammalian cells may be cultured cells or cell lines, for example. Representative mammalian cells include lymphoid cell lines (e.g., NSO), Chinese hamster ovary cells (CHO), human Per C6 cell lines (e.g., PER C6 cells from Crucell), COS cells, oocytes, and cells from transgenic animals, e.g., mammary epithelial cells.

[0111] In one aspect, the present invention relates to a method for providing an antibody molecule as described herein. The method comprises providing a LAG-3 antigen (e.g., an antigen comprising at least a portion of a LAG-3 epitope); obtaining an antibody molecule that specifically binds to a LAG-3 polypeptide; and evaluating whether the antibody molecule specifically binds to the LAG-3 polypeptide or evaluating the efficacy of the antibody molecule in modulating, for example, inhibiting, LAG-3 activity. The method may further comprise administering the antibody molecule to a target, for example, a human or a non-human animal.

[0112] In other aspects, the present invention provides compositions, such as pharmaceutical compositions, comprising a pharmaceutically acceptable carrier, additive or stabilizer and at least one of the anti-LAG3 antibody molecules described herein. In one embodiment, the composition, such as pharmaceutical compositions, comprises an anti-LAG-3 antibody molecule and one or more agents described herein, such as a therapeutic agent or a combination of other antibody molecules. In one embodiment, the antibody molecule is conjugated with a label or therapeutic agent.

[0113] The antibody molecules disclosed herein can inhibit, reduce, or neutralize one or more activities of LAG-3. In one embodiment, the anti-LAG-3 antibody molecule is CD4 + T lymphocytes or CD8 + Increased antigen-dependent stimulation of T lymphocytes, increased T cell proliferation; increased expression of activating antigens, e.g., CD25; increased expression of cytokines, e.g., interferon-gamma (IFN-γ), interleukin-2 (IL-2), interleukin-4 (IL-4), chemokine (CC motif) ligand 3 (CCL3), chemokine (CC motif) ligand 4 (CCL4), or chemokine (CC motif) ligand 5 (CCL5); T reg This results in one or more of the following: decreased cellular suppressor activity, increased T cell homeostasis, increased tumor-infiltrating lymphocytes, or decreased immune evasion by cancer cells. Therefore, such antibody molecules can be used alone or in combination to treat and prevent disorders in which enhancement of the immune response in the target is desired.

[0114] Use of anti-LAG-3 antibody molecules Therefore, in other respects, methods for modulating the immune response in a subject are provided. These methods involve administering an antibody molecule disclosed herein (e.g., a therapeutically effective dose of an anti-LAG-3 antibody molecule) alone or in combination with one or more agents or methods, so as to modulate the immune response in a subject. In some embodiments, the antibody molecule restores, enhances, stimulates, or increases the immune response in the subject.

[0115] The subjects may be mammals, e.g., primates, preferably higher primates, e.g., humans (e.g., patients with or at risk of having the disorders described herein). In some embodiments, the subjects have a need for an enhanced immune response. In some embodiments, the anti-LAG-3 antibody molecule restores, enhances, or stimulates the antigen-specific T cell response in the subject, e.g., interleukin-2 (IL-2) or interferon-gamma (IFN-γ) production. In some embodiments, the immune response is an antitumor response. In some embodiments, the subjects have or are at risk of having the disorders described herein, e.g., cancer or infectious disorders described herein. In some embodiments, the subjects are immunocompromised or at risk of becoming immunocompromised. For example, the subjects are receiving or have received chemotherapy and / or radiotherapy in the past. Separately or in combination with this, the subjects are immunocompromised or at risk of becoming immunocompromised as a result of infection.

[0116] In some respects, the present invention provides a method for treating cancer or tumors in a subject (e.g., one or more of mitigation, inhibition, or progression delay). The method comprises administering to the subject a therapeutically effective dose of an anti-LAG-3 antibody molecule as described herein, e.g., an anti-LAG-3 antibody molecule, alone, e.g., as monotherapy, or in combination with one or more agents or methods. In some embodiments, the anti-LAG-3 antibody molecule is administered in combination with a co-inhibitor modulator (e.g., a co-inhibitor agonist) or an inhibitory molecule modulator (e.g., an immune checkpoint inhibitor), e.g., as described herein. In some embodiments, the anti-LAG-3 antibody molecule is administered in combination with an immune checkpoint modulator inhibitor or activator (e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule), a TIM-3 modulator (e.g., a TIM-3 activator or inhibitor, e.g., an anti-TIM-3 antibody molecule), or a CTLA-4 inhibitor (e.g., an anti-CTLA4 antibody).

[0117] In one embodiment, cancers treated with anti-LAG-3 antibody molecules, alone or in combination, include solid tumors, hematopoietic cancers (e.g., leukemia, lymphoma, myeloma) and their metastatic lesions. In one embodiment, cancer is a solid tumor. Examples of solid tumors include malignancies of diverse organ systems, such as those affecting the lungs, breasts, lymphatic system, digestive or colorectal tract, reproductive and urogenital tract (e.g., kidneys, urothelium, bladder cells), pharynx, CNS (e.g., brain, nerve or glial cells), skin (e.g., melanoma), head and neck (e.g., head and neck squamous cell carcinoma (HNCC)), and pancreas. For example, this includes melanoma, colon cancer, gastric cancer, rectal cancer, renal cell carcinoma, breast cancer (e.g., breast cancer that does not express estrogen receptor, progesterone receptor or Her2 / neu 1, 2 or all of them, e.g., trinegative breast cancer), liver cancer, lung cancer (e.g., non-small cell lung cancer (NSCLC) (e.g., NSCLC with squamous and / or non-squamous histology) or small cell lung cancer), prostate cancer, head and neck cancer (e.g., HPV-positive squamous cell carcinoma), small intestine cancer, and esophageal cancer. Examples of hematopoietic cancers include, but are not limited to, leukemia (e.g., myeloid leukemia, lymphocytic leukemia, or chronic lymphocytic leukemia (CLL)), lymphoma (e.g., Hodgkin lymphoma (HL), non-Hodgkin lymphoma (NHL), disseminated large B-cell lymphoma (DLBCL), T-cell lymphoma, or mantle cell lymphoma (MCL)), and myeloma, such as multiple myeloma. Cancer can be early, intermediate, late, or metastatic.

[0118] In one embodiment, cancer is selected from colorectal cancer (e.g., CRC), melanoma, e.g., advanced melanoma (e.g., stage II-IV melanoma) or HLA-A2 positive melanoma; pancreatic cancer, e.g., advanced pancreatic cancer; breast cancer, e.g., metastatic breast cancer or trinegative breast cancer; head and neck cancer (e.g., HNSCC); esophageal cancer; renal cell carcinoma (RCC), e.g., clear renal cell carcinoma (ccRCC) or metastatic renal cell carcinoma (MRCC); lung cancer (e.g., NSCLC); cervical cancer; bladder cancer; or hematological malignancies, e.g., leukemia (e.g., lymphocytic leukemia) or lymphoma (e.g., Hodgkin lymphoma (HL), non-Hodgkin lymphoma (NHL), generalized large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL) or CLL, e.g., relapsed or refractory chronic lymphocytic leukemia).

[0119] In one embodiment, the cancer is advanced or unresectable melanoma that does not respond to other treatments. In another embodiment, the cancer is melanoma with a BRAF mutation (e.g., BRAF V600 mutation). In yet another embodiment, the anti-LAG-3 antibody molecule is used alone (e.g., as monotherapy) or in combination with one or more secondary agents (e.g., BRAF inhibitors). In one embodiment, the anti-LAG-3 antibody molecule is administered (e.g., before, after, or concurrently with) an immune checkpoint modulator inhibitor (e.g., PD-1 inhibitor, PD-L1 inhibitor, TIM-3 inhibitor, CEACAM (e.g., CEACAM1 and / or CEACAM5) inhibitor or CTLA4 inhibitor (e.g., anti-CLA4 antibody, e.g., ipilimumab)) in combination with or without a BRAF inhibitor (e.g., vemurafenib or dabrafenib) to treat the melanoma. In one embodiment, an anti-LAG-3 antibody molecule is administered in combination with a PD-1 or PD-L1 inhibitor, such as an anti-PD-1 or anti-PD-L1 antibody molecule, to treat melanoma as described herein.

[0120] In one embodiment, an anti-LAG-3 antibody molecule is administered alone, for example as monotherapy, or in combination with an immune checkpoint modulator inhibitor (e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule), a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule), a CEACAM (e.g., a CEACAM1 and / or CEACAM5) inhibitor (e.g., an anti-CEACAM antibody molecule) or a CTLA-4 inhibitor (e.g., an anti-CTLA4 antibody) to treat head and neck cancers such as those described herein.

[0121] In one embodiment, an anti-LAG-3 antibody molecule is administered alone, for example as monotherapy, or in combination with an immune checkpoint modulator inhibitor or activator (e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule), a TIM-3 modulator (e.g., a TIM-3 activator or inhibitor, for example, an anti-TIM-3 antibody molecule), a CEACAM (e.g., a CEACAM1 and / or CEACAM5) inhibitor (e.g., an anti-CEACAM antibody molecule), or a CTLA-4 inhibitor (e.g., an anti-CTLA4 antibody) for the treatment of lung cancer (e.g., NSCLC) as described herein.

[0122] In one embodiment, an anti-LAG-3 antibody molecule is administered alone, for example as monotherapy, or in combination with an immune checkpoint modulator inhibitor (e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule), a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule), a CEACAM (e.g., a CEACAM1 and / or CEACAM5) inhibitor (e.g., an anti-CEACAM antibody molecule) or a CTLA-4 inhibitor (e.g., an anti-CTLA4 antibody) to treat gastric cancer as described herein. In one embodiment, an anti-LAG-3 antibody molecule is administered in combination with a PD-1 or PD-L1 inhibitor, for example, an anti-PD-1 or anti-PD-L1 antibody molecule, to treat gastric cancer as described herein.

[0123] In one embodiment, an anti-LAG-3 antibody molecule may be used alone, for example as monotherapy, or as an inhibitor of an immune checkpoint modulator (for example, a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), or a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule) to treat lymphomas (e.g., Hodgkin lymphoma (HL), non-Hodgkin lymphoma (NHL), disseminated large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL), or CLL, e.g., relapsed or refractory chronic lymphocytic leukemia) The anti-LAG-3 antibody molecule is administered in combination with a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule), a CEACAM (e.g., CEACAM1 and / or CEACAM5) inhibitor (e.g., an anti-CEACAM antibody molecule), or a CTLA-4 inhibitor (e.g., an anti-CTLA4 antibody). In one embodiment, the anti-LAG-3 antibody molecule is administered in combination with a PD-1 or PD-L1 inhibitor, e.g., an anti-PD-1 or anti-PD-L1 antibody molecule, to treat lymphomas such as those described herein.

[0124] In one embodiment, the tumor microenvironment has elevated PD-L1 expression levels. Separately or in combination with this, the tumor microenvironment may have elevated IFNγ and / or CD8 expression.

[0125] In one embodiment, an anti-LAG-3 antibody molecule is administered alone or in combination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) or a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule) to treat subjects identified as having or having tumors that are high PD-L1 levels or expression and TIL+ (e.g., having an increased TIL count) or both. In one embodiment, a subject is identified as having or having tumors that are high PD-L1 levels or expression and TIL+. In one embodiment, the method described herein further includes identifying a subject based on having a tumor that is high PD-L1 levels or expression and TIL+ or one or more of both. In one embodiment, the method described herein further includes identifying a subject based on having a tumor that is high PD-L1 levels or expression and TIL+. In one embodiment, a TIL+ tumor is CD8 and IFNγ positive. In one embodiment, a subject is identified as having or possessing a high percentage of cells that are positive for one or more of PD-L1, CD8, and / or IFN-γ. In another embodiment, a subject is identified as having or possessing a high percentage of cells that are positive for all of PD-L1, CD8, and IFN-γ.

[0126] In one embodiment, the method described herein further includes identifying subjects based on a high percentage of cells positive for one or more PD-L1, CD8, and / or IFN-γ. In one embodiment, the method described herein further includes identifying subjects based on a high percentage of cells positive for all of PD-L1, CD8, and / or IFN-γ. In one embodiment, subjects are identified as having or having one, two, or more PD-L1, CD8, and / or IFN-γ and one or more lung cancers, e.g., squamous cell lung cancer or lung adenocarcinoma, head and neck cancer, squamous cell cervical cancer, gastric cancer, esophageal cancer, thyroid cancer, melanoma, and / or nasopharyngeal cancer (NPC). In one embodiment, the method described herein further describes identifying subjects based on having PD-L1, CD8 and / or IFN-γ 1, 2 or more and one or more lung cancers, such as squamous cell lung cancer or lung adenocarcinoma, head and neck cancer, squamous cell cervical cancer, gastric cancer, thyroid cancer, melanoma and / or nasopharyngeal cancer.

[0127] The methods and compositions described herein are useful for treating metastatic lesions associated with the aforementioned cancer.

[0128] In a further aspect, the present invention provides a method for treating an infectious disease in a subject, comprising administering to the subject a therapeutically effective dose of the anti-LAG-3 antibody molecule described herein, either alone or in combination with one or more agents or methods. While the antibody of the present invention is preferred for use in this method, other anti-LAG-3 antibodies or their antigen-binding fragments may be used instead (or in combination with the anti-LAG-3 antibody molecule described herein).

[0129] In one embodiment, the infectious disease is hepatitis (e.g., hepatitis B infection). In one embodiment, an anti-LAG-3 antibody molecule is administered in combination with a hepatitis B antigen or vaccine and optionally with an aluminum-containing adjuvant.

[0130] In another embodiment, the infectious disease is influenza. In one embodiment, an anti-LAG-3 antibody molecule is administered in combination with an influenza antigen or vaccine.

[0131] Furthermore, the present invention provides a method for enhancing the immune response to an antigen in a subject, comprising administering to the subject (i) an antigen; and (ii) an anti-LAG-3 antibody molecule, such that the immune response to the antigen in the subject is enhanced. The antigen may be, for example, a tumor antigen, a viral antigen, a bacterial antigen, or an antigen from a pathogen.

[0132] Anti-LAG-3 antibody molecules can be administered to a subject systemically (e.g., orally, non-enteral, subcutaneous, intravenously, rectally, intramuscularly, intraperitoneally, intranasally, percutaneously, or by inhalation or intracavitary placement), topically, or by application to mucous membranes such as the nose, throat, and bronchi, either alone or in combination.

[0133] The dosage and treatment regimen of the anti-LAG-3 antibody molecule can be determined by those skilled in the art. In one embodiment, the anti-LAG-3 antibody molecule is administered by injection (e.g., subcutaneously or intravenously) in doses of approximately 1 to 30 mg / kg, for example, approximately 5 to 25 mg / kg, approximately 10 to 20 mg / kg, approximately 1 to 10 mg / kg, or approximately 1 mg / kg, 3 mg / kg, or 10 mg / kg. The dosing schedule can vary, for example, from once a week to once every two, three, or four weeks. In one embodiment, the anti-LAG-3 antibody molecule is administered every other week at a dose of approximately 10 to 20 mg / kg. In one embodiment, the anti-LAG-3 antibody molecule is administered (e.g., intravenously) in doses of approximately 3 to 800 mg, for example, approximately 3 mg, 20 mg, 80 mg, 240 mg, or 800 mg. In one embodiment, the anti-LAG-3 antibody molecule is administered alone in doses of approximately 20 to 800 mg, for example, approximately 3 mg, 20 mg, 80 mg, 240 mg, or 800 mg. In another embodiment, the anti-LAG-3 antibody molecule is administered in doses of approximately 3 to 240 mg, for example, approximately 3 mg, 20 mg, 80 mg, or 240 mg, when combined with a second agent or therapeutic modality, for example, one of the second agents or therapeutic modalities described herein. In one embodiment, the anti-LAG-3 antibody molecule is administered every two weeks (for example, between weeks 1, 3, 5, and 7) during each 8-week cycle up to 96 weeks.

[0134] The antibody molecules described herein are suitable for use in the methods described herein, but other anti-LAG-3 antibodies can be used in place of or in combination with the anti-LAG-3 antibody molecules of the present invention.

[0135] Combination therapy The methods and compositions described herein can be used in combination with other agents or treatments. In one embodiment, the method described herein includes administering the anti-LAG-3 antibody molecule described herein in an amount effective for treating or preventing the disorder, in combination with an agent, treatment, or modality. The anti-LAG-3 antibody molecule and the agent, treatment, or modality can be administered simultaneously or sequentially in any order. Any combination and order of the anti-LAG-3 antibody molecule and other therapeutic agents, methods, or modalities (e.g., those described herein) can be used. The antibody molecule and / or other therapeutic agents, methods, or modalities can be administered during periods of active disorder or during periods of remission or low-activity disease. The antibody molecule can be administered before, concurrently with, after, or during remission of the disorder.

[0136] In one embodiment, the methods and compositions described herein are administered in combination with one or more of the following: other antibody molecules, chemotherapeutic agents, other anticancer agents (e.g., targeted anticancer agents, gene therapy, viral therapy, RNA therapy, bone marrow transplantation, nanotherapy, or oncolytic agents), cytotoxic agents, immuno-based therapies (e.g., cytokine or cell-based immunotherapy), surgical procedures (e.g., mammary tumor removal or mastectomy), and / or radiotherapy or any combination thereof. Further treatment may take the form of adjuvant or neoadjuvant therapy. In one embodiment, further treatment may be an enzyme inhibitor (e.g., a small molecule enzyme inhibitor) or a metastasis inhibitor.

[0137] Examples of typical cytotoxic agents that can be administered concomitantly include microtubule inhibitors, topoisomerase inhibitors, antimetabolites, mitotic inhibitors, alkylating agents, anthracyclines, vinca alkaloids, interstitials, agents that can interfere with signaling pathways, agents that promote apoptosis, proteosome inhibitors, and irradiation (e.g., local or total body irradiation (e.g., gamma irradiation)). In other embodiments, further treatment is surgery or irradiation or a combination thereof. In other embodiments, further treatment involves one or more of the PI3K / AKT / mTOR pathways. The targeted therapies are HSP90 inhibitors or tubulin inhibitors. Other typical antibody molecules that can be administered in combination include, but are not limited to, checkpoint inhibitors (e.g., anti-PD-1, anti-PD-L1); antibodies that stimulate immune cells (e.g., agonist GITR or CD137 antibody); and anticancer antibodies (e.g., rituximab (Rituxan® or MabThera®), trastuzumab (Herceptin®), cetuximab (Erbitux®)).

[0138] In addition to or in combination with the aforementioned combinations, the methods and compositions described herein may be administered in combination with one or more immunomodulators (e.g., activators of costimulatory molecules or inhibitors of immunosuppressive molecules, e.g., immune checkpoint molecules); vaccines, e.g., therapeutic cancer vaccines; or other forms of cellular immunotherapy.

[0139] Representative, non-limiting combinations and uses of anti-LAG-3 antibody molecules include the following:

[0140] In one embodiment, an anti-LAG-3 antibody molecule is administered in combination with a modulator of a co-inhibitory molecule (e.g., an agonist of a co-inhibitory molecule) or a modulator of an inhibitory molecule (e.g., an inhibitor of an immune checkpoint inhibitor).

[0141] In one embodiment, an anti-LAG-3 antibody molecule is administered in combination with a co-inhibitor modulator, such as an agonist. In one embodiment, the co-inhibitor agonist is selected from agonists (e.g., agonist antibodies or soluble fusions) of OX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a / CD18), ICOS (CD278), 4-1BB (CD137), GITR, CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160, B7-H3, or CD83 ligands.

[0142] In one embodiment, an anti-LAG-3 antibody molecule is administered in combination with an inhibitor of an inhibitory (or immune checkpoint) molecule selected from PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4, CEACAM (e.g., CEACAM-1 and / or CEACAM-5) and / or TGFR beta. Inhibition of the inhibitory molecule may be carried out by inhibition at the DNA, RNA, or protein level. In one embodiment, an inhibitory nucleic acid (e.g., dsRNA, siRNA, or shRNA) can be used to inhibit the expression of the inhibitory molecule. In another embodiment, the inhibitor of the inhibitory signal is a polypeptide that binds to the inhibitory molecule, e.g., a soluble ligand or antibody or antibody fragment. In one embodiment, the inhibitor is a soluble ligand (e.g., CTLA-4-Ig) or antibody or antibody fragment that binds to PD-1, PD-L1, PD-L2, or CTLA-4.

[0143] For example, administering an anti-LAG-3 antibody molecule in combination with an inhibitor of PD-1, PD-L1, PD-L2, or CTLA-4, such as an antibody or antibody fragment that binds to it, can help treat cancer (e.g., colorectal cancer (e.g., CRC); melanoma, such as advanced melanoma (e.g., stage II-IV melanoma) or HLA-A2 positive melanoma; pancreatic cancer, such as advanced pancreatic cancer; breast cancer, such as metastatic breast cancer or triglyceride-negative breast cancer; head and neck cancer (e.g., HNSCC); esophageal cancer; renal cell carcinoma) It can treat (RCC), for example, clear renal cell carcinoma (ccRCC) or metastatic renal cell carcinoma (MRCC); lung cancer (e.g., NSCLC); cervical cancer; bladder cancer; or hematological malignancies, for example, leukemia (e.g., lymphocytic leukemia) or lymphoma (e.g., Hodgkin lymphoma (HL), non-Hodgkin lymphoma (NHL), generalized large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL), or CLL, for example, cancer selected from relapsed or refractory chronic lymphocytic leukemia).

[0144] In one embodiment, an anti-LAG-3-1 antibody molecule is administered in combination with (e.g., before, together with, or after) treatment with an anti-CTLA4 antibody (e.g., ipilimumab), with or without a BRAF inhibitor (e.g., vemurafenib or dabrafenib).

[0145] In another embodiment, the anti-LAG-3 antibody molecule is administered in combination with an anti-PD-1 antibody (e.g., nivolumab or pembrolizumab) or its antigen-binding fragment. In yet another embodiment, the anti-LAG-3 antibody molecule is administered in combination with an anti-TIM-3 antibody or its antigen-binding fragment. In yet another embodiment, the anti-LAG-3 antibody molecule is administered in combination with an anti-PD-L1 antibody or its antigen-binding fragment. In yet another embodiment, the anti-LAG-3 antibody molecule is administered in combination with an anti-PD-1 antibody and an anti-TIM-3 antibody (or its antigen-binding fragment). In one embodiment, the anti-LAG-3 antibody molecule is administered in combination with an anti-PD-1 antibody and an anti-PD-L1 antibody (or its antigen-binding fragment). In one embodiment, the anti-LAG-3 antibody molecule is administered in combination with an anti-TIM-3 antibody and an anti-PD-L1 antibody (or its antigen-binding fragment).

[0146] In another embodiment, the anti-LAG-3 antibody molecule is administered in combination with a CEACAM inhibitor (e.g., a CEACAM-1 and / or CEACAM-5 inhibitor), such as an anti-CEACAM antibody molecule. In another embodiment, the anti-LAG-3 antibody molecule is administered in combination with a CEACAM-1 inhibitor, such as an anti-CEACAM-1 antibody molecule. In another embodiment, the anti-LAG-3 antibody molecule is administered in combination with a CEACAM-5 inhibitor, such as an anti-CEACAM-5 antibody molecule.

[0147] In yet another embodiment, an anti-LAG-3 antibody molecule is administered in combination with an anti-CEACAM (e.g., anti-CEACAM-1 and / or anti-CEACAM-5) antibody molecule and an anti-PD-1 antibody molecule. In yet another embodiment, an anti-LAG-3 antibody molecule is administered in combination with an anti-CEACAM (e.g., anti-CEACAM-1 and / or anti-CEACAM-5) antibody molecule and an anti-TIM-3 antibody molecule. In yet another embodiment, an anti-LAG-3 antibody molecule is administered in combination with an anti-CEACAM (e.g., anti-CEACAM-1 and / or anti-CEACAM-5) antibody molecule and an anti-PD-L1 antibody molecule. The antibody combinations described herein may be administered separately, for example, as individual antibodies or their antigen-binding fragments, or conjugated together, for example, as bispecific or trispecific antibody molecules. In one embodiment, a bispecific antibody comprising an anti-LAG-3 antibody molecule and one of the following is administered: an anti-TIM-3 antibody, an anti-CEACAM (e.g., anti-CEACAM-1 and / or anti-CEACAM-5) antibody, an anti-PD-L1 antibody, or an anti-PD-1 antibody or its antigen-binding fragment. In one embodiment, the antibody combination described herein is used to treat cancer, for example, cancers such as those described herein (e.g., solid tumors or hematological malignancies). In one embodiment, the anti-LAG-3 antibody molecule is administered in combination with an anti-PD-1 or anti-PD-L1 antibody to treat solid tumors.

[0148] In another embodiment, an anti-LAG-3 antibody molecule is administered in combination with a cytokine. The cytokine can be administered as a fusion molecule to the anti-LAG-3 antibody molecule or as a separate composition. In one embodiment, the anti-LAG-3 antibody is administered in combination with one, two, three or more cytokines, for example, as a fusion molecule or as a separate composition. In one embodiment, the cytokine is an IL selected from one, two, three or more interleukins (IL)-1, IL-2, IL-12, IL-15, or IL-21. In one embodiment, the bispecific antibody molecule has a first binding specificity to a first target (e.g., LAG-3) and a second binding specificity to a second target (e.g., PD-1, TIM-3, or PD-L1), and optionally binds to an interleukin (e.g., IL-12) domain, e.g., full-length IL-12 or a portion thereof. In one embodiment, the anti-LAG-3 antibody molecule and cytokine combination described herein is used for the treatment of cancer, e.g., cancers such as those described herein (e.g., solid tumors).

[0149] In other embodiments, an anti-LAG-3 antibody molecule is administered in combination with a vaccine, e.g., a therapeutic cancer vaccine or another form of cellular immunotherapy. In some embodiments, the vaccine may be peptide-based, DNA-based, RNA-based, antigen-based, or a combination thereof. In some embodiments, the vaccine comprises one or more peptides, nucleic acids (e.g., DNA or RNA), antigens, or a combination thereof. In some embodiments, the cancer vaccine comprises an adjuvant (e.g., aluminum phosphate or aluminum hydroxide). In some embodiments, the methods described herein are administered in combination with one or more surgical excision of tissue, chemotherapy, or other anti-cancer treatments, the primary or sole target being metastatic lesions, e.g., metastases to the bone marrow or lymph nodes.

[0150] In one embodiment, the cancer is melanoma, e.g., advanced melanoma (e.g., stage II-IV melanoma) or HLA-A2 positive melanoma. In one embodiment, an anti-LAG-3 antibody molecule is combined with a tumor antigenic peptide, e.g., one or more HLA-A2 peptides, and optionally with an adjuvant, e.g., Montanide. TM It is administered in combination with the anti-LAG-3 antibody molecule. Representative onco-peptides that can be administered in combination with the anti-LAG-3 antibody molecule include tyrosinase.A2, MAGE-C2.A2, NY-ESO-1b.A2, MAGE-4.A2, MAGE-3.A2, MAGE-1.A2, NA17.A2(GnTV), and MAGE-10.A2.

[0151] In another embodiment, the cancer is pancreatic cancer, for example, advanced pancreatic cancer. In one embodiment, the antibody molecule can be administered in combination with a chemotherapeutic agent, for example, gemcitabine.

[0152] In another embodiment, the cancer is breast cancer, for example, metastatic breast cancer or 3-negative breast cancer. In one embodiment, the antibody molecule can be administered in combination with a chemotherapeutic agent, for example, paclitaxel.

[0153] In other embodiments, cancer is renal cell carcinoma, for example, advanced clear cell carcinoma (e.g., stage IV) or metastatic renal cell carcinoma (MRCC). In another aspect, the cancer is a head and neck cancer, such as HPV-positive squamous cell carcinoma.

[0154] In another embodiment, the anti-LAG-3 antibody molecule is administered in combination with an antigen. For example, the anti-LAG-3 antibody molecule is administered in combination with a hepatitis B antigen (e.g., Engerix B). In another embodiment, the anti-LAG-3 antibody molecule is administered in combination with an influenza antigen.

[0155] Anti-LAG-3 antibody molecules may be used alone in a non-conjugated form, or they may be conjugated to substances, such as cytotoxic agents or moieties (e.g., therapeutic drugs; radioactive compounds; molecules of plant, fungal, or bacterial origin; or biological proteins (e.g., protein toxins) or particles (e.g., recombinant viral particles via viral coat proteins). For example, antibodies can be coupled to radioactive isotopes such as α-, β-, or γ-emitters or β- and γ-emitters.

[0156] Further combination therapy The methods and compositions described herein (e.g., LAG-3 antibodies and their uses) can be used in combination with other drugs or therapies, such as a second therapeutic agent selected from one or more drugs listed in Table 7. In one embodiment, the method described herein comprises administering to a subject the anti-LAG-3 antibody molecule described herein (optionally PD-1, PD-L1, TIM-3, CEACAM (e.g., in combination with one or more inhibitors of CEACAM-1 and / or CEACAM-5) or CTLA-4), and further comprising administering a second therapeutic agent selected from one or more drugs listed in Table 7 in an amount effective for treating or preventing a disorder, such as the disorder described herein, such as cancer. When administered in combination, the anti-LAG-3 antibody molecule, the additional drug (e.g., a second or third drug), or all of them can be administered individually, for example, in amounts or doses higher, lower, or the same as the amount or dose of each drug used as a monotherapy. In one embodiment, the amount or dose of the anti-LAG-3 antibody, additional agents (e.g., a second or third agent), or all administered is lower individually (e.g., at least 20%, at least 30%, at least 40%, or at least 50%) than the amount or dose of each agent used as monotherapy. In another embodiment, the amount or dose of the anti-LAG-3 antibody, additional agents (e.g., a second or third agent), or all administered is lower (e.g., at least 20%, at least 30%, at least 40%, or at least 50%) to produce the desired effect (e.g., treatment of cancer).

[0157] In other embodiments, the second therapeutic agent is selected from one or more of the drugs listed in Table 7. In some embodiments, the cancer is selected from lung cancer (e.g., non-small cell lung cancer (NSCLC) (e.g., NSCLC or NSCLC adenocarcinoma with squamous and / or non-squamous histology)) or as disclosed in the publications listed in Table 7.In one embodiment, the second therapeutic agent is, for example, as described here and in Table 7, 1) protein kinase C (PKC) inhibitors; 2) heat shock protein 90 (HSP90) inhibitors; 3) phosphoinositide 3-kinase (PI3K) and / or target of rapamycin (mTOR) inhibitors; 4) cytochrome P450 inhibitors (e.g., CYP17 inhibitors or 17-alpha-hydroxylase / C17-20 lyase inhibitors); 5) iron chelators; 6) aromatase inhibitors; 7) p5 3) Inhibitors, e.g., p53 / Mdm2 interaction inhibitors; 8) Apoptosis inducers; 9) Angiogenesis inhibitors; 10) Aldosterone synthase inhibitors; 11) Smound (SMO) receptor inhibitors; 12) Prolactin receptor (PRLR) inhibitors; 13) Wnt signaling inhibitors; 14) CDK4 / 6 inhibitors; 15) Fibroblast growth factor receptor 2 (FGFR2) / Fibroblast growth factor receptor 4 (FGFR4) inhibitors; 16) Macrophage colony-stimulating factor (M-CSF) inhibitors; 17) One or more inhibitors of c-KIT, histamine release, Flt3 (e.g., FLK2 / STK1), or PKC; 18) One or more inhibitors of VEGFR-2 (e.g., FLK-1 / KDR), PDGFR beta, c-KIT, or Raf kinase C; 19) Somatostatin agonists and / or growth hormone release inhibitors; 20) Anaplastic lymphoma kinase (ALK) inhibitors; 21) Insulin-like growth factor 1 receptor (IGF-1R) inhibitors; 22) P-glycoprotein 1 inhibitors; 23) Vascular One or more of the following are selected: 24) Endothelial growth factor receptor (VEGFR) inhibitors; 25) BCR-ABL kinase inhibitors; 26) FGFR inhibitors; 27) CYP11B2 inhibitors; 28) HDM2 inhibitors, e.g., HDM2-p53 interaction inhibitors; 29) Tyrosine kinase inhibitors; 30) c-MET inhibitors; 31) JAK inhibitors; 32) DAC inhibitors; 33) 11β-hydroxylase inhibitors; 34) IAP inhibitors; 35) PIM kinase inhibitors; 36) Porcupine inhibitors; 37) BRAF inhibitors, e.g., BRAF V600E or wild-type BRAF inhibitors; 38) HER3 inhibitors; 39) MEK inhibitors; or lipid kinase inhibitors.

[0158] In one embodiment, the second therapeutic agent is selected from one or more of compounds A8, A17, A23, A24, A27, A29, A33, and A13.

[0159] In another embodiment, the second therapeutic agent is selected from one or more compounds A5, A8, A17, A23, A24, A29, and A40.

[0160] In another embodiment, the second therapeutic agent is selected from one or more compounds A9, A16, A17, A21, A22, A25, A28, A48, and 49.

[0161] In one embodiment, the second therapeutic agent is administered at a therapeutic dose or a dose lower than therapeutic. In one embodiment, the concentration of the second therapeutic agent required to achieve inhibition, e.g., proliferation inhibition, is lower when the second therapeutic agent is combined with an anti-LAG-3 antibody molecule than when the second therapeutic agent is administered separately. In one embodiment, the concentration of the anti-LAG-3 antibody molecule required to achieve inhibition, e.g., proliferation inhibition, is lower when the anti-LAG-3 antibody molecule is administered in combination with the second therapeutic agent than when the anti-LAG-3 antibody molecule is administered separately. In one embodiment, in combination therapy, the concentration of the second therapeutic agent required to achieve inhibition, e.g., proliferation inhibition, is lower than the therapeutic dose of the second therapeutic agent as monotherapy, e.g., 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, or 80-90% lower. In one embodiment, in combination therapy, the concentration of the anti-LAG-3 antibody molecule required to achieve inhibition, e.g., proliferation inhibition, is lower than the therapeutic dose of the anti-PD-1 antibody molecule as monotherapy, e.g., 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, or 80-90% lower.

[0162] detection In other words, the present invention relates to a method for detecting the presence of LAG-3 in a sample, e.g., in vitro or in vivo (e.g., a biological sample, e.g., serum, semen or urine, or e.g., a tissue biopsy from a hyperproliferative or cancerous lesion). The method can be used for evaluation (e.g., treatment or progression, diagnosis and / or monitoring of the disorders described herein in a subject, e.g., hyperproliferative or cancerous disorders). The method comprises (i) contacting a sample (and optionally a reference, e.g., a control sample) with the anti-LAG-3 antibody molecule described herein under conditions that induce interaction, or administering it to a subject, and (ii) detecting the formation of a complex between the antibody molecule and the sample (and optionally a reference, e.g., a control sample). The formation of the complex is an indicator of LAG-3 and may indicate the suitability or necessity of the treatment described herein. The method may include immunohistochemistry, immunocytochemistry, flow cytometry (e.g., FACS), antibody molecule complexing magnetic beads, ELISA assay, and PCR (e.g., RT-PCR).

[0163] Generally, antibody molecules used in in vivo and in vitro diagnostic methods are directly or indirectly labeled with detectable substances that facilitate the detection of bound or unbound binders. Suitable detectable substances include a variety of biologically active enzymes, prosthetic groups, fluorescent substances, luminescent substances, paramagnetic (e.g., nuclear magnetic resonance active) substances, and radioactive substances.

[0164] Further embodiments provide a method for treating cancer, comprising identifying the presence of 1, 2, or all of PD-L1, CD8, or IFN-γ in a sample (e.g., a sample of a subject containing cancer cells and optionally immune cells such as TILs), thereby providing values ​​for 1, 2, or all of PD-L1, CD8, and IFN-γ. The method further comprises comparing the PD-L1, CD8, and / or IFN-γ values ​​to a reference value, e.g., a control value, and if the PD-L1, CD8, and / or IFN-γ values ​​are higher than the reference value, e.g., a control value, administering a therapeutically effective dose of an anti-LAG-3 antibody (e.g., the anti-LAG-3 antibody described herein) to the subject alone or in combination with an anti-PD-1 antibody molecule, an anti-PD-L1 antibody molecule, or both, and optionally in combination with one or more other agents, thereby treating the cancer. Cancer may be any of the cancers described herein, such as, for example, lung cancer (squamous cell), lung cancer (adenocarcinoma), head and neck cancer, cervical cancer (squamous cell), gastric cancer, thyroid cancer, melanoma, nasopharyngeal cancer, or breast cancer, such as TN breast cancer, such as IM-TN breast cancer. In some aspects, cancer may be ER+ breast cancer or pancreatic cancer.

[0165] Also provided is a method for treating cancer, comprising testing a sample (e.g., a sample of a subject containing cancer cells) for the presence of PD-L1, thereby identifying the PD-L1 value, comparing the PD-L1 value to a control value, and if the PD-L1 value is higher than the control value, administering a therapeutically effective dose of an anti-LAG-3 antibody (e.g., the anti-LAG-3 antibody described herein) to the subject alone or in combination with an anti-PD-1 antibody molecule, an anti-PD-L1 antibody molecule, or both, or optionally in combination with one or more other agents, thereby treating the cancer. The cancer is, for example, one of the cancers described herein, which may be, for example, non-small cell lung (NSCLC) adenocarcinoma (ACA), NSCLC squamous cell carcinoma (SCC), or hepatocellular carcinoma (HCC).

[0166] In other respects, the present invention relates to a diagnostic or therapeutic kit comprising the anti-LAG-3 antibody molecule described herein and instructions for its use.

[0167] All publications, patent applications, patents, and other documents mentioned herein are incorporated herein by reference in their entirety.

[0168] Other characteristics, purposes, and advantages of the present invention are evident from the specification and drawings, as well as from the claims. [Brief explanation of the drawing]

[0169] [Figure 1] Figure 1 shows the amino acid sequences of the light chain (SEQ ID NO: 16) and heavy chain (SEQ ID NO: 6) variable regions of the mouse anti-LAG-3 mAb BAP050. The light chain and heavy chain CDR sequences based on Kabat numbering are underlined. The light chain and heavy chain CDR sequences based on Chothia numbering are shown in bold italics.

[0170] [Figure 2] Figure 2 shows the amino acid sequences of the light chain (SEQ ID NO: 16) and heavy chain (SEQ ID NO: 6) variable regions of mouse anti-LAG-3 mAb BAP050 aligned with germline sequences (SEQ ID NOs: 290-291, in order of appearance). The upper and lower sequences are the germline (GL) and BAP050 (Mu mAb) sequences, respectively. Light and heavy chain CDR sequences based on Kabat numbering are underlined. Light and heavy chain CDR sequences based on Chothia numbering are shown in bold italics. "-" indicates identical amino acid residues.

[0171] [Figure 3] Figure 3 is a bar graph showing the results of FACS binding analysis of 20 humanized BAP050 clones (BAP050-hum01 to BAP050-hum20) and a chimeric mAb (BAP050-chi). The antibody concentrations for each tested mAb are 200 ng / ml, 100 ng / ml, 50 ng / ml, 25 ng / ml, and 12.5 ng / ml from the leftmost bar to the rightmost bar.

[0172] [Figure 4]Figure 4 shows the structural analysis of humanized BAP049 clones (a, b, c, d, e, f, g represent various types of framework region sequences). The concentrations of mAbs in the samples are also shown.

[0173] [Figure 5] Figures 5A-5B show the binding affinity and specificity of humanized mAbs measured in a competitive binding assay using a constant concentration of FITC-labeled mouse mAbs, serial dilutions of the test antibody, and LAG-3-expressing CHO cells. The experiment was performed twice, and the results are shown in Figures 5A and 5B, respectively.

[0174] [Figure 6] Figure 6 shows the ranking of humanized BAP050 clones based on FACS data, competitive binding, and structural analysis. The concentrations of mAbs in the samples are also shown.

[0175] [Figure 7] Figure 7 shows the binding affinity and specificity of huBAP050(Ser) clones as measured in a competitive binding assay using a constant concentration of FITC-labeled mouse mAb, serial dilutions of the test antibody, and LAG-3 expressing CHO cells. HuBAP050(Ser) clones such as BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum05-Ser, BAP050-hum09-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, and BAP050-hum13-Ser were evaluated. Mouse mAb BAP050, chimeric mAb BAP050-chi, and humanized BAP050-hum01, BAP050-hum02, BAP050-hum05, BAP050-hum09, BAP050-hum11, BAP050-hum12, and BAP050-hum13 were also included in this analysis.

[0176] [Figure 8]Figure 8 shows the blockade of LAG-3-Ig binding to Daudi cells by the huBAP050(Ser) clone. HuBAP050(Ser) clones such as BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum05-Ser, BAP050-hum09-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, and BAP050-hum13-Ser were evaluated. Mouse mAb BAP050 and chimeric mAb BAP050-chi were also included in this analysis.

[0177] [Figure 9A] Figures 9A and 9B show the alignment of the heavy chain variable domain sequences of 20 humanized BAP050 clones and BAP050 chimeras (BAP050-chi). Figure 9A shows all sequences (in order of appearance, SEQ ID NOs. 20, 28, 28, 28, 28, 28, 28, 28, 28, 64, 64, 64, 64, 64, 68, 72, 72, 76, and 80). Figure 9B shows only the amino acid sequences that differ from the mouse sequences (in order of appearance, SEQ ID NOs. 20, 28, 28, 28, 28, 28, 28, 28, 28, 28, 64, 64, 64, 64, 64, 68, 72, 72, 76, and 80). [Figure 9B] Figures 9A and 9B show the alignment of the heavy chain variable domain sequences of 20 humanized BAP050 clones and BAP050 chimeras (BAP050-chi). Figure 9A shows all sequences (in order of appearance, SEQ ID NOs. 20, 28, 28, 28, 28, 28, 28, 28, 28, 64, 64, 64, 64, 64, 68, 72, 72, 76, and 80). Figure 9B shows only the amino acid sequences that differ from the mouse sequences (in order of appearance, SEQ ID NOs. 20, 28, 28, 28, 28, 28, 28, 28, 28, 28, 64, 64, 64, 64, 64, 68, 72, 72, 76, and 80).

[0178] [Figure 10A]Figures 10A and 10B show the alignment of the light chain variable domain sequences of 20 humanized BAP050 clones and BAP050 chimeras (BAP050-chi). Figure 10A shows all sequences (in order of appearance, SEQ ID NOs. 24, 32, 36, 36, 36, 292, 292, 292, 44, 48, 52, 56, 56, 60, 60, 60, 60, 84, 88, 92, and 96). Figure 10B shows only the amino acid sequences that differ from the mouse sequences (in order of appearance, SEQ ID NOs. 24, 32, 36, 36, 36, 292, 292, 292, 44, 48, 52, 56, 56, 60, 60, 60, 60, 84, 88, 92, and 96). [Figure 10B] Figures 10A and 10B show the alignment of the light chain variable domain sequences of 20 humanized BAP050 clones and BAP050 chimeras (BAP050-chi). Figure 10A shows all sequences (in order of appearance, SEQ ID NOs. 24, 32, 36, 36, 36, 292, 292, 292, 44, 48, 52, 56, 56, 60, 60, 60, 60, 84, 88, 92, and 96). Figure 10B shows only the amino acid sequences that differ from the mouse sequences (in order of appearance, SEQ ID NOs. 24, 32, 36, 36, 36, 292, 292, 292, 44, 48, 52, 56, 56, 60, 60, 60, 60, 84, 88, 92, and 96).

[0179] [Figure 11] Figure 11 shows representative cancers with a relatively high proportion of patients who are positive for all three tests: PD-L1, CD8, and IFN-γ.

[0180] [Figure 12] Figure 12 shows representative ER+ breast and pancreatic cancers with a relatively low proportion of patients who are positive for all three PD-L1 / CD8 / IFN-γ tests.

[0181] [Figure 13] Figure 13 shows the proportion of representative breast cancer patients who are positive for all three PD-L1 / CD8 / IFN-γ tests.

[0182] [Figure 14] Figure 14 shows the proportion of representative colon cancer patients who are positive for all three PD-L1 / CD8 / IFN-γ tests.

[0183] A brief explanation of the table Table 1 is a summary of the amino acid and nucleotide sequences of mouse, chimeric, and humanized anti-LAG-3 antibody molecules. The antibody molecules include mouse mAb BAP050 and chimeric mAb BAP050-chi, humanized mAb BAP050-hum01 to BAP050-hum20, BAP050-hum01-Ser to BAP050-hum15-Ser, BAP050-hum18-Ser to BAP050-hum20-Ser, and BAP050-Clone-F to BAP050-Clone-J. The amino acid and nucleotide sequences of the heavy chain and light chain CDR, the amino acid and nucleotide sequences of the heavy chain and light chain variable regions, and the amino acid and nucleotide sequences of the heavy chain and light chain are shown in this table.

[0184] Table 2 shows the amino acid and nucleotide sequences of the heavy and light chain framework regions of the humanized mAbs BAP050-hum01 to BAP049-hum20, BAP050-hum01-Ser to BAP050-hum15-Ser, BAP050-hum18-Ser to BAP050-hum20-Ser, and BAP049-Clone-F to BAP049-Clone-J.

[0185] Table 3 shows the constant region amino acid sequences of the human IgG heavy chain and human kappa light chain. Table 4 shows the amino acid sequences of the heavy chain and light chain leader sequences of the humanized mAb BAP050-Clone-F to BAP050-Clone-J.

[0186] Table 5 summarizes the yield, titer, monomer content, and endotoxin levels of representative humanized BAP050 mAbs expressed in CHO cells. Table 6 shows the charge isoforms of representative humanized BAP050 expressed in CHO cells, as detected by Novex IEF analysis.

[0187] Table 7 is a summary of selected therapeutic agents that can be administered in combination with the anti-LAG-3 antibody molecules described herein and other immunomodulators (e.g., one or more co-inhibitor activators and / or immune checkpoint molecule inhibitors). From left to right, Table 7 lists the compound name, compound structure, and patent numbers disclosing the compound for each second therapeutic agent. [Modes for carrying out the invention]

[0188] Detailed description The immune system has the ability to recognize and eliminate tumor cells; however, tumors can employ numerous strategies to evade immunity. Blocking immune checkpoints is one attempt to activate or reactivate therapeutic antitumor immunity. Lymphocyte-activating gene-3 (LAG-3) has been described as an inhibitory receptor in immune synapses (Chen and Flies (2013) Nat Rev Immunol. 13(4):227-42). Therefore, blocking LAG-3 may lead to an enhancement of antitumor immunity.

[0189] Several cell types express LAG-3. For example, LAG-3 is activated CD4 + and CD8 + T cells, T reg It is expressed in cells, natural killer (NK) cells, and plasmacytoid dendritic cells (DCs). LAG-3 is expressed in tumor-infiltrating lymphocytes, for example, in infiltrating lymphocytes in head and neck squamous cell carcinoma (HNSCC). LAG-3 is expressed in highly suppressed induced T reg and natural T reg It is expressed in, for example, highly inhibitory FoxP3 + nT reg and FoxP3 - iT regLAG-3 is positive in melanoma and colorectal cancer (Camisaschi et al. (2010) J. Immunol. 184(11):6545-6551; Scurr et al. (2014) Mucosal. Immunol. 7(2):428-439).

[0190] LAG-3 negatively regulates T cell signaling and function. Ligands for LAG-3 include, for example, MHC class II and L-SECtin. Anti-LSECtin has been shown to inhibit B16 melanoma cell proliferation (Xu et al. (2014) Cancer Res. 74(13):3418-3428). Blocking LAG-3 restores the activity of effector cells, T reg This can reduce suppressor activity and / or enhance anti-PD-1 antitumor activity.

[0191] LAG-3 is generally PD-1 + Although not exclusively co-expressed on cells, blocking a single IFN-γ / TNF-α molecule can restore the cell's in vitro activity. For example, CD8 is shown by the percentage of dual IFN-γ / TNF-α producing strains. + The degree of T cell exhaustion correlates with the number of inhibitory receptors expressed (Blackburn et al. (2009) Nat. Immunol. 10(1): 29-37). High PD-1 / LAG-3 expression correlates with T cell infiltration in melanoma. Co-blockade of LAG-3 and anti-PD-1 or PD-L1 may produce tumor suppressor activity in preclinical models. For example, anti-LAG-3 and anti-PD-1 blockade has shown efficacy in Sa1N fibrosarcoma and MC38 colon cancer models (Woo et al. (2012) Cancer Res. 72(4):917-27).

[0192] LAG-3 blockade is also effective in lymphocytic choriomeningitis virus (LCMV) models. For example, PD-L1+ LAG-3 blockade during chronic LCMV infection is effective against CD8 +It enhances the T cell response (Blackburn et al. (2009) Nat. Immunol. 10(1): 29-37).

[0193] Accordingly, the present invention relates, at least in part, to antibody molecules (e.g., humanized antibody molecules) that bind to lymphocyte-activating gene-3 (LAG-3) with high affinity and specificity. In one embodiment, a humanized antibody against LAG-3 exhibiting low immunogenicity is disclosed. For example, the humanized BAP050 antibody was found to have risk scores of less than 1200, 1150, 1100, 1050, 1000, 950, 900, 850, or 800 in the T cell epitope assay described herein. In other embodiments, selected combinations of framework regions, such as those shown in Figures 4 and 6, have been shown to have different production efficiencies and binding characteristics.

[0194] Further aspects of the present invention include nucleic acid molecules encoding antibody molecules, expression vectors, host cells, and methods for producing antibody molecules. Pharmaceutical compositions comprising immune complexes, multi- or bispecific molecules, and antibody molecules are also provided. The anti-LAG-3 antibody molecules disclosed herein can be used for the treatment, prevention, and / or diagnosis of cancerous or malignant disorders (e.g., melanoma, e.g., advanced melanoma; pancreatic cancer, e.g., advanced pancreatic cancer; solid tumors; breast cancer, e.g., metastatic breast cancer; renal cell carcinoma, e.g., cancers such as advanced or metastatic renal cell carcinoma (MRCC) or clear cell renal cell carcinoma), as well as infectious diseases (e.g., hepatitis, e.g., hepatitis B; influenza). Therefore, methods for detecting LAG-3, and methods for treating a variety of disorders, including cancer and infectious diseases, using anti-LAG-3 antibody molecules alone or in combination are disclosed herein.

[0195] The term "lymphocyte activation gene-3" or "LAG-3" includes all isoforms, mammals, such as human LAG-3, species homologs of human LAG-3, and analogs containing at least one common epitope with LAG-3. The amino acid and nucleotide sequences of LAG-3, such as human LAG-3, are known in the art (e.g., Triebel et al. (1990) J. Exp. Med. 171:1393-1405).

[0196] Additional terms are defined below and in the specification. As used herein, the singular forms refer to the grammatical object of the article being one or more (e.g., at least one).

[0197] The term "or" means "and / or" herein and is used interchangeably with it, unless the context clearly indicates otherwise.

[0198] "About" and "approximately" generally mean the degree of allowable error of the measured quantity, taking into account the nature or accuracy of the measurement. Representative degrees of error are within 20 percent (%) of a value or range of values, generally within 10%, and more commonly within 5%.

[0199] The compositions and methods of the present invention include polypeptides and nucleic acids having sequences that are identical or substantially identical or similar to the specified sequences, e.g., sequences that are at least 70%, 75%, 80%, 85%, 90%, 95% or more identical to the specified sequences. In amino acid sequences, the term "substantially identical" is used herein to refer to a first amino acid that is i) identical or ii) contains a sufficient or minimum number of amino acid residues such that the aligned amino acid residues in a second amino acid sequence are conservative substitutions such that the first and second amino acid sequences may have a common structural domain and / or a common functional activity. For example, an amino acid sequence containing a common structural domain having at least about 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to a target sequence, such as the sequences provided herein.

[0200] In nucleotide sequences, the term “substantially identical” is used to mean a first nucleic acid sequence containing a sufficient or minimum number of nucleotides that are identical to the aligned nucleotides in the second nucleic acid sequence such that the first and second nucleotide sequences encode a polypeptide having common functional activity or encode a common structural polypeptide domain or common functional polypeptide activity. For example, the target sequence, e.g., a nucleotide sequence having at least about 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the sequence provided herein.

[0201] The term "functional variant" refers to a polypeptide that has substantially the same amino acid sequence as a naturally occurring sequence, or is encoded by substantially the same nucleotide sequence, and is capable of having one or more of the activities of a naturally occurring sequence.

[0202] The calculation of homology or sequence identity between sequences (these terms are used interchangeably here) is performed as follows:

[0203] To determine the percentage of identity between two amino acid sequences or two nucleic acid sequences, these sequences are aligned for optimal comparison purposes (for example, gaps can be inserted into one or both of the first and second amino acid or nucleic acid sequences for optimal alignment, and non-homologous sequences can be ignored for comparison purposes). In a preferred embodiment, the length of the target sequences aligned for comparison purposes is at least 30%, preferably at least 40%, more preferably at least 50%, 60%, and even more preferably at least 70%, 80%, 90%, or 100% of the length of the target sequences. The amino acid residues or nucleotides at the corresponding amino acid or nucleotide positions are then compared. If the position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical in that position (as used herein, amino acid or nucleic acid "identity" is equivalent to amino acid or nucleic acid "homology").

[0204] The identity percentage between two arrays is a function of the number of identical positions shared by the arrays, taking into account the number of gaps that needed to be inserted for optimal alignment of the two arrays and the length of each gap.

[0205] The comparison of sequences and the determination of the percentage of identity between two sequences can be achieved using mathematical algorithms. In a preferred embodiment, the percentage of identity between two amino acid sequences is determined using the Needleman and Wunsch ((1970) J. Mol. Biol. 48:444-453)) algorithm, which is incorporated into the GAP program of the GCG software package (available at http: / / www.gcg.com), using a Blossum 62 matrix or PAM250 matrix and gap weights of 16, 14, 12, 10, 8, 6, or 4 and length weights of 1, 2, 3, 4, 5, or 6. In yet another preferred embodiment, the percentage of identity between two nucleotide sequences is determined using the GAP program of the GCG software package (available at http: / / www.gcg.com), using an NWSgapdna.CMP matrix and gap weights of 40, 50, 60, 70, or 80 and length weights of 1, 2, 3, 4, 5, or 6. A particularly preferred set of parameters (and one that should be used unless otherwise specified) is the Blossum 62 scoring matrix with a gap penalty of 12, a gap extension penalty of 4, and a frameshift gap penalty of 5.

[0206] The percentage of identity between two amino acid or nucleotide sequences can be determined using the algorithm by E. Meyers and W. Miller ((1989) CABIOS, 4:11-17), which is incorporated into the ALIGN program (version 2.0), using the PAM120 weighted residue table, gap length penalty 12, and gap penalty 4.

[0207] The nucleic acid and protein sequences described herein can be used as “query sequences” to perform searches against public databases, for example, to identify other family members or related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) described in Altschul, et al. (1990) J. Mol. Biol. 215:403-10. A BLAST nucleotide search can be performed in the NBLAST program with a score of 100 and a word length of 12 to obtain nucleotide sequences homologous to the nucleic acid (SEQ ID NO: 1) molecule of the present invention. A BLAST protein search can be performed in the XBLAST program with a score of 50 and a word length of 3 to obtain amino acid sequences homologous to the protein molecule of the present invention. For comparative purposes, gap alignment can be obtained using Gapped BLAST as described in Altschul et al., (1997) Nucleic Acids Res. 25:3389-3402. When using the BLAST and Gapped BLAST programs, you can use the default parameters for each program (e.g., XBLAST and NBLAST). See http: / / www.ncbi.nlm.nih.gov.

[0208] The terms “hybridize under low stringency, medium stringency, high stringency, or very high stringency conditions” used herein describe the hybridization and washing conditions. Guidelines for carrying out the hybridization reaction can be found in Current Protocols in Molecular Biology, John Wiley & Sons, NY (1989), 6.3.1–6.3.6, which are incorporated herein by reference. Aqueous and non-aqueous methods are described in that literature, and either can be used. The specific hybridization conditions referred to herein are as follows: 1) Low stringency hybridization conditions are two washes with 6× sodium chloride / sodium citrate (SSC), 45°C, followed by 0.2× SSC, 0.1% SDS at at least 50°C (wash temperature may be raised to 55°C for low stringency conditions); 2) Medium stringency hybridization conditions are one or more washes with 6× SSC, 45°C, followed by 0.2× SSC, 0.1% SDS at 60°C; 3) High stringency hybridization conditions are one or more washes with 6× SSC, about 45°C, followed by 0.2× SSC, 0.1% SDS at 65°C; and preferably 4) Ultra-high stringency hybridization conditions are one or more washes with 0.5M sodium phosphate, 7% SDS, 65°C, followed by 0.2× SSC, 1% SDS, 65°C. Very high stringency conditions (4) are preferred conditions and should be used unless otherwise specified.

[0209] It is understood that the molecules of the present invention may have further conservative or non-essential amino acid substitutions that do not substantially affect their function.

[0210] The term “amino acid” is intended to encompass all molecules, whether natural or synthetic, that include both amino- and acid-functional properties and can be included in polymers of naturally occurring amino acids. Representative amino acids include naturally occurring amino acids; their analogs, derivatives, and congeners; amino acid analogs with heteromorphic side chains; and all stereoisomers of any of the above. The term “amino acid” as used herein includes both D- and L-optical isomers and peptide mimetic compounds.

[0211] A “conservative amino acid substitution” is one in which an amino acid residue is replaced by an amino acid residue having a similar side chain. Families of amino acid residues with similar side chains are defined in this art. These families include amino acids having basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).

[0212] The terms “polypeptide,” “peptide,” and “protein” (if single-chain) are used interchangeably here to refer to polymers of amino acids of any length. Polymers may be linear or branched, may contain modified amino acids, or may be interrupted by non-amino acids. The term also encompasses amino acid polymers that have undergone any other modification, such as disulfide bond formation, glycosylation, lipid addition, acetylation, phosphorylation, or attachment of labeling elements. Polypeptides can be isolated from natural sources, produced by recombinant techniques from eukaryotic or prokaryotic hosts, or are products of synthetic methods.

[0213] The terms “nucleic acid,” “nucleic acid sequence,” “nucleotide sequence,” or “polynucleotide sequence,” and “polynucleotide” are interchangeable. A polynucleotide is a polymeric form of nucleotides of any length, either deoxyribonucleotides or ribonucleotides or their analogues. A polynucleotide may be single-stranded or double-stranded, and if single-stranded, it may be a coding strand or a non-coding (antisense) strand. A polynucleotide may contain modified nucleotides such as methylated nucleotides and nucleotide analogs. The nucleotide sequence may be interrupted by non-nucleotide elements. Polynucleotides may be further assembled after polymerization, such as in conjugation with a labeling component. Nucleic acids may be recombinant polynucleotides or polynucleotides of genomic, cDNA, semi-synthetic, or synthetic origin, which are not naturally occurring or are conjugated with other polynucleotides in unnatural configurations.

[0214] The term “isolated” as used herein refers to a substance that has been removed from its original or natural environment (for example, its natural environment if it exists naturally). For example, a naturally occurring polynucleotide or polypeptide present in a living animal is not isolated, but the same polynucleotide or polypeptide separated by human intervention from some or all of the substances that coexist in the natural system is isolated. Such a polynucleotide is part of a vector and / or such a polynucleotide or polypeptide is part of a composition, and such a vector or composition is still isolated in that it is not part of the environment in which it naturally occurs.

[0215] Various aspects of the present invention are described in further detail below. Further definitions are given in the specification.

[0216] antibody molecule In certain embodiments, the antibody molecule binds to mammalian, e.g., human LAG-3. For example, the antibody molecule specifically binds to an epitope of LAG-3, e.g., a linear or conformational epitope (e.g., an epitope described herein). In certain embodiments, the antibody molecule binds to one or more extracellular Ig-like domains of LAG-3, e.g., the first, second, third, or fourth extracellular Ig-like domain of LAG-3.

[0217] As used herein, the term "antibody molecule" refers to a protein comprising at least one immunoglobulin variable domain sequence, e.g., an immunoglobulin chain or a fragment thereof. The term "antibody molecule" includes, for example, monoclonal antibodies (including full-length antibodies having an immunoglobulin Fc region). In certain embodiments, the antibody molecule comprises a full-length antibody or a full-length immunoglobulin chain. In certain embodiments, the antibody molecule comprises an antigen-binding or functional fragment of a full-length antibody or a full-length immunoglobulin chain.

[0218] In certain embodiments, the antibody molecule is a monospecific antibody molecule that binds to a single epitope. For example, a monospecific antibody molecule having multiple immunoglobulin variable domain sequences that each bind to the same epitope.

[0219] In one embodiment, the antibody molecule is a polyspecific antibody molecule, for example, comprising a number of immunoglobulin variable domain sequences, where the number of first immunoglobulin variable domain sequences has binding specificity to a first epitope and the number of second immunoglobulin variable domain sequences has binding specificity to a second epitope. In one embodiment, the first and second epitopes are on the same antigen, for example, the same protein (or a subunit of a multimeric protein). In one embodiment, the first and second epitopes overlap. In one embodiment, the first and second epitopes do not overlap. In one embodiment, the first and second epitopes are on different antigens, for example, different proteins (or different subunits of a multimeric protein). In one embodiment, the polyspecific antibody molecule comprises a third, fourth, or fifth immunoglobulin variable domain. In one embodiment, the polyspecific antibody molecule is a bispecific antibody molecule, a triplicate antibody molecule, or a quadruplicate antibody molecule.

[0220] In some embodiments, a polyspecific antibody molecule is a bispecific antibody molecule. A bispecific antibody does not have specificity for more than two antigens. A bispecific antibody molecule is characterized by a first immunoglobulin variable domain sequence having binding specificity to a first epitope and a second immunoglobulin variable domain sequence having binding specificity to a second epitope. In some embodiments, the first and second epitopes are on the same antigen, e.g., the same protein (or a subunit of a multimeric protein). In some embodiments, the first and second epitopes overlap. In some embodiments, the first and second epitopes do not overlap. In some embodiments, the first and second epitopes are on different antigens, e.g., different proteins (or different subunits of a multimeric protein). In some embodiments, a bispecific antibody molecule comprises a heavy chain variable domain sequence and a light chain variable domain sequence having binding specificity to a first epitope and a heavy chain variable domain sequence and a light chain variable domain sequence having binding specificity to a second epitope. In some embodiments, a bispecific antibody molecule comprises a semi-antibody having binding specificity to a first epitope and a semi-antibody having binding specificity to a second epitope. In one embodiment, the bispecific antibody molecule comprises a semi-antibody or a fragment thereof having binding specificity to a first epitope and a semi-antibody or a fragment thereof having binding specificity to a second epitope. In another embodiment, the bispecific antibody molecule comprises an scFv or a fragment thereof having binding specificity to a first epitope and an scFv or a fragment thereof having binding specificity to a second epitope. In another embodiment, the first epitope is located on LAG-3, and the second epitope is located on PD-1, TIM-3, CEACAM (e.g., CEACAM-1 and / or CEACAM-5), PD-L1, or PD-L2.

[0221] In one embodiment, an antibody molecule comprises a bispecific antibody and a single-chain molecule, as well as antigen-binding fragments of the antibody (e.g., Fab, F(ab')2, and Fv). For example, an antibody molecule may comprise a heavy (H) chain variable domain sequence (hereinafter abbreviated as VH) and a light (L) chain variable domain sequence (hereinafter abbreviated as VL). In one embodiment, an antibody molecule comprises or consists of one heavy chain and one light chain (hereinafter referred to as a half-antibody). In other examples, an antibody molecule comprises two heavy (H) chain variable domain sequences and two light (L) chain variable domain sequences, thereby forming two antigen-binding sites such as Fab, Fab', F(ab')2, Fc, Fd, Fd', Fv, a single-chain antibody (e.g., scFv), a monovariate domain antibody, a bispecific antibody (Dab) (bivalent and bispecific), and a chimeric (e.g., humanized) antibody, which may be produced by modification of a complete antibody or de novo synthesized using recombinant DNA technology. These functional antibody fragments retain their selective binding ability to their respective antigens or receptors. Antibodies and antibody fragments may be derived from any class and any subclass of antibodies (e.g., IgG1, IgG2, IgG3, and IgG4), including but not limited to IgG, IgA, IgM, IgD, and IgE. Preparations of antibody molecules may be monoclonal or polyclonal. Antibody molecules may also be human, humanized, CDR-transplanted, or in vitro-produced antibodies. Antibodies may have a heavy chain constant region selected from, for example, IgG1, IgG2, IgG3, or IgG4. Antibodies may also have a light chain selected from, for example, kappa or lambda. The term “immunoglobulin” (Ig) is used here interchangeably with the term “antibody.”

[0222] Examples of antigen-binding fragments of antibody molecules include (i) monovalent fragments consisting of Fab fragments, VL, VH, CL, and CH1 domains; (ii) bivalent fragments containing two Fab fragments linked by disulfide crosslinks in the hinge region, F(ab')2 fragments; (iii) Fd fragments consisting of VH and CH1 domains; (iv) Fv fragments consisting of VL and VH domains of one arm of the antibody; (v) bispecific antibody (dAb) fragments consisting of a VH domain; (vi) camelid or camelid variable domains; (vii) single-stranded Fv (scFv) (see, e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883); and (viii) single-domain antibodies. These fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened for their usefulness in the same manner as complete antibodies.

[0223] The term “antibody” includes the complete molecule and its functional fragments. The constant region of an antibody may be modified, for example, by mutation, to alter its properties (e.g., for Fc receptor binding, antibody glycosylation, cysteine ​​residue count, or an increase or decrease of one or more effector cell functions or complement functions).

[0224] The antibodies of the present invention may also be single-domain antibodies. Single-domain antibodies may include antibodies in which the complementarity-determining region is part of a single-domain polypeptide. Examples include, but are not limited to, heavy-chain antibodies, naturally light-chain-deficient antibodies, conventionally four-chain-derived single-domain antibodies, manipulated antibodies, and single-domain scaffolds other than those derived from antibodies. Single-domain antibodies may be any cutting-edge or future single-domain antibodies. Single-domain antibodies may be derived from any species, including, but not limited to, mice, humans, camels, llamas, fish, sharks, goats, rabbits, and cattle. In other aspects of the present invention, single-domain antibodies are naturally occurring single-domain antibodies known as light-chain-deficient heavy-chain antibodies. Such single-domain antibodies are disclosed, for example, in WO94 / 04678. For clarity, the variable domain derived from this naturally light-chain-deficient heavy-chain antibody is known here as VHH or nanobody to distinguish it from VH of conventional four-chain immunoglobulins. Such VHH molecules may originate from antibodies induced in camelid species, such as camels, llamas, dromedaries, alpacas, and guanacos. Other species outside the camelid family can naturally produce heavy-chain antibodies lacking light chains, and such VHHs fall within the scope of this invention.

[0225] VH and VL regions can be subdivided into more conserved regions called "framework regions" (FR or FW) and hypervariable regions called "complementarity-determining regions" (CDR). The degree of framework regions and CDR is precisely defined by many methods (see Kabat, EA, et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242; Chothia, C. et al. (1987) J. Mol. Biol. 196:901-917; and AbM definition using Oxford Molecular's AbM antibody modeling software). Generally, see, for example, Protein Sequence and Structure Analysis of Antibody Variable Domains. In: Antibody Engineering Lab Manual (Ed.: Duebel, S. and Kontermann, R., Springer-Verlag, Heidelberg).

[0226] The terms "complementarity-determining region" and "CDR" used here refer to the amino acid sequences within the antibody variable region that confer antigen specificity and binding affinity. Generally, each heavy chain variable region has three CDRs (HCDR1, HCDR2, HCDR3), and each light chain variable region has three CDRs (LCDR1, LCDR2, LCDR3).

[0227] The precise amino acid sequence boundaries of a certain CDR are defined in Kabat et al. (1991), “Sequences of Proteins of Immunological Interest,” 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (“Kabat” numbering scheme), and Al-Lazikani et al., (1997) JMB 273, 927-948 (“Chothia” numbering scheme). The CDRs defined using the “Chothia” numbering scheme are sometimes called “highly variable loops.”

[0228] For example, Kabat numbered the CDR amino acid residues in the heavy chain variable domain (VH) as 31-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3), and the CDR amino acid residues in the light chain variable domain (VL) as 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3). Chothia numbered the CDR amino acids in VH as 26-32 (HCDR1), 52-56 (HCDR2), and 95-102 (HCDR3), and the amino acid residues in VL as 26-32 (LCDR1), 50-52 (LCDR2), and 91-96 (LCDR3). By combining the Kabat and Chothia definitions of CDRs, the CDR in human VH consists of amino acid residues 26-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3), while the CDR in human VL consists of amino acid residues 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3).

[0229] Generally, unless otherwise specified, anti-LAG-3 antibody molecules are combinations of Kabat CDRs and / or Chothia highly variable loops, for example, as listed in Table 1. In one embodiment, the following definitions are used for the anti-LAG-3 antibody molecules listed in Table 1: HCDR1 by a combination of the CDR definitions of both Kabat and Chothia, and HCCDR2-3 and LCCDR1-3 by the CDR definition of Kabat. In all definitions, each VH and VL generally contains 6 CDRs and 4 FRs arranged in the following order from amino-terminus to carboxyl-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0230] As used herein, “immunoglobulin variable domain sequence” refers to an amino acid sequence capable of forming the structure of an immunoglobulin variable domain. For example, the sequence may include all or part of the amino acid sequence of a naturally occurring variable domain. For example, the sequence may or may not include one, two, or more N-terminal or C-terminal amino acids, or may include other modifications that are compatible with the formation of the protein structure.

[0231] The term “antigen-binding site” refers to the portion of an antibody molecule that contains determinants that form an interface for binding to the LAG-3 polypeptide or its epitope. For proteins (or protein mimes), the antigen-binding site generally comprises one or more loops (at least four amino acids or amino acid mimes) that form an interface for binding to the LAG-3 polypeptide. Generally, the antigen-binding site of an antibody molecule comprises at least one or two CDRs and / or highly variable loops, or more typically at least three, four, five, or six CDRs and / or highly variable loops.

[0232] The terms “competition” or “cross-competition” are used interchangeably here to refer to the ability of an antibody molecule to interfere with the binding of an anti-LAG-3 antibody molecule, e.g., the target of the anti-LAG-3 antibody molecule provided herein, e.g., human LAG-3. The interference with binding may be direct or indirect (e.g., via allosteric regulation of the antibody molecule or target). The extent to which an antibody molecule can interfere with the binding of another antibody molecule to its target, and therefore whether or not it constitutes competition, can be determined using competitive binding assays, e.g., FACS assays, ELISAs, or BIACORE assays. In some embodiments, the competitive binding assay is a quantitative competitive assay. In one embodiment, a first anti-LAG-3 antibody molecule is considered competitive when, in a competitive binding assay (for example, the competitive assay described herein), the binding of the first antibody molecule to the target decreases by 10% or more, for example, 20%, 30%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% or more.

[0233] The term “epitope” as used herein refers to a portion of an antigen (e.g., human LAG-3) that specifically interacts with an antibody molecule. Such portions, referred to here as epitope determinants, generally constitute or are part of elements such as amino acid side chains or sugar side chains. Epitope determinants can be determined by methods known in the art or disclosed herein, e.g., crystallography or hydrogen-deuterium exchange. At least one or more portions on the antibody molecule that specifically interact with epitope determinants are generally located in the CDR. Generally, epitopes have specific three-dimensional structural features. Generally, epitopes have specific charge features. Some epitopes are linear epitopes, while others are stereoepitopes.

[0234] The terms “monoclonal antibody” or “monoclonal antibody composition” as used herein refer to preparations of antibody molecules in single-molecule composition form. Monoclonal antibody compositions exhibit single-binding specificity and affinity for a particular epitope. Monoclonal antibodies can be manufactured by hybridoma technology or by non-hybridoma technology methods (e.g., recombinant methods).

[0235] An “effective human” protein is a protein that does not induce a neutralizing antibody response, such as a human anti-mouse antibody (HAMA) response. HAMA can be problematic in many situations, for example, when antibody molecules are repeatedly administered, for example, in the treatment of chronic or relapsing disease conditions. The HAMA response may render repeated antibody administration ineffective due to increased antibody clearance from serum (see, e.g., Saleh et al., Cancer Immunol. Immunother., 32:180-190 (1990)) and also due to the possibility of an allergic reaction (see, e.g., LoBuglio et al., Hybridoma, 5:5117-5123 (1986)).

[0236] The antibody molecule may be either a polyclonal antibody or a monoclonal antibody. In other embodiments, the antibody can be produced by recombinant methods, for example, by phage display or combinatorial methods.

[0237] Phage display and combinatorial methods for antibody production are well known in this field (e.g., Ladner et al. U.S. Patent No. 5,223,409; Kang et al. International Publication WO92 / 18619; Dower et al. International Publication WO91 / 17271; Winter et al. International Publication WO92 / 20791; Markland et al. International Publication WO92 / 15679; Breitling et al. International Publication WO93 / 01288; McCafferty et al. International Publication WO92 / 01047; Garrard et al. International Publication WO92 / 09690; Ladner et al. International Publication WO90 / 02809; Fuchs et al. (1991) Bio / Technology 9:1370-1372; Hay et al. (1992)). Hum Antibod Hybridomas 3:81-85; Huse et al. (1989) Science 246:1275-1281; Griffths et al. (1993) EMBO J 12:725-734; Hawkins et al. (1992) J Mol Biol 226:889-896; Clackson et al. (1991) Nature 352:624-628; Gram et al. (1992) PNAS 89:3576-3580; Garrad et al. (1991) Bio / Technology 9:1373-1377; Hoogenboom et al. (1991) Nuc Acid Res 19:4133-4137; and Barbas et al. (1991) PNAS As disclosed in 88:7978-7982 (these contents are incorporated herein by reference).

[0238] In one embodiment, the antibody may be a fully human antibody (e.g., an antibody produced in a mouse genetically engineered to produce antibodies from a human immunoglobulin sequence) or a non-human antibody, such as a rodent (mouse or rat), goat, primate (e.g., monkey), or camel antibody. Preferably, the non-human antibody is a rodent (mouse or rat antibody). Methods for producing rodent antibodies are known in the art.

[0239] Human monoclonal antibodies can be produced using transgenic mice that carry human immunoglobulin genes, rather than using mouse strains. Splenocytes derived from these transgenic mice immunized with the target antigen are used to produce hybridomas that secrete human mAbs with specific affinity for epitopes from human proteins (e.g., Wood et al. International Application WO91 / 00906, Kucherlapati et al. PCT Publication WO91 / 10741; Lonberg et al. International Application WO92 / 03918; Kay et al. International Application 92 / 03917; Lonberg, N. et al. 1994 Nature 368:856-859; Green, LL et al. 1994 Nature Genet. 7:13-21; Morrison, SL et al. 1994 Proc. Natl. Acad. Sci. USA 81:6851-6855; Bruggeman et al. 1993 Year Immunol 7:33-40; Tuaillon et al. 1993 PNAS 90:3720-3724; Bruggeman et al. 1991 Eur J Immunol 21:1323-1326).

[0240] Antibodies may have a variable region or part thereof, for example, a CDR, produced in a non-human organism, such as a rat or mouse. Chimeric antibodies, CDR-implanted antibodies, and humanized antibodies are within the scope of the present invention. Antibodies produced in a non-human organism, such as a rat or mouse, and then modified, for example, within the variable framework or constant region, to reduce antigenicity in humans are also within the scope of the present invention.

[0241] Chimeric antibodies can be produced by recombinant DNA methods known in this field (Robinson et al., International Patent Application PCT / US86 / 02269; Akira, et al., European Patent Applications 184,187; Taniguchi, M., European Patent Application 171,496; Morrison et al., European Patent Application 173,494; Neuberger et al., International Application WO86 / 01533; Cabilly et al., US Patent No. 4,816,567; Cabilly et al., European Patent Application 125,023; Better et al. (1988 Science 240:1041-1043); Liu et al. (1987) PNAS 84:3439-3443; Liu et al., 1987, J. Immunol. 139:3521-3526; Sun et al. See also: al. (1987) PNAS 84:214-218; Nishimura et al., 1987, Canc. Res. 47:999-1005; Wood et al. (1985) Nature 314:446-449; and Shaw et al., 1988, J. Natl Cancer Inst. 80:1553-1559).

[0242] Humanized or CDR-transplanted antibodies have at least one or two, but generally three, recipient CDRs (heavy and / or light immunoglobulin chains) that have been replaced by donor CDRs. The antibody may have at least some of the non-human CDRs replaced, or only some of the CDRs may have been replaced with non-human CDRs. It is only necessary to replace the number of CDRs required for the humanized antibody to bind to LAG-3. Preferably, the donor is a rodent antibody, e.g., a rat antibody or a mouse antibody, and the recipient is a human framework or human consensus framework. Generally, the immunoglobulin that provides the CDRs is called the “donor,” and the immunoglobulin that provides the framework is called the “acceptor.” In some embodiments, the donor immunoglobulin is non-human (e.g., rodent). The acceptor framework is a naturally occurring (e.g., human) framework or consensus framework or a sequence that matches it by about 85% or more, preferably 90%, 95%, 99%, or more.

[0243] The term “consensus sequence” as used herein refers to a sequence formed from the most frequently occurring amino acids (or nucleotides) in a family of related sequences (see, for example, Winnaker, From Genes to Clones (Verlagsgesellschaft, Weinheim, Germany 1987)). In a family of proteins, each position in the consensus sequence is occupied by the most frequently occurring amino acid at that position in the family. If two amino acids occur equally frequently, both can be included in the consensus sequence. “Consensus framework” refers to the framework region in a consensus immunoglobulin sequence.

[0244] Antibodies can be humanized using methods known in this field (see Morrison, SL, 1985, Science 229:1202-1207, by Oi et al., 1986, BioTechniques 4:214 and Queen et al., US issues 5,585,089, 5,693,761, and 5,693,762).

[0245] Humanized or CDR-implanted antibodies can substitute 1, 2, or all CDRs in the immunoglobulin chain, and can be produced by CDR implantation or CDR substitution. See, for example, U.S. Patent No. 225,539; Jones et al. 1986 Nature 321:552-525; Verhoeyan et al. 1988 Science 239:1534; Beidler et al. 1988 J. Immunol. 141:4053-4060; Winter U.S. Patent No. 5,225,539 (all of which are expressly incorporated herein by reference). Winter describes a CDR implantation method that can be used to produce the humanized antibodies of the present invention (U.S. Patent Application GB2188638A (filed March 26, 1987); Winter U.S. Patent No. 5,225,539), which is expressly incorporated herein by reference.

[0246] Furthermore, the scope of this invention includes humanized antibodies in which specific amino acids are substituted, deleted, or added. Criteria for selecting amino acids from a donor are described, for example, in columns 12-16 of U.S. Patent No. 5,585,089, which are incorporated herein by reference. Other techniques for humanizing antibodies are described in Padlan et al. EP519596A1, published December 23, 1992.

[0247] The antibody molecule may be a single-chain antibody. Single-chain antibodies (scFVs) can be manipulated (see, for example, Colcher, D. et al. (1999) Ann NY Acad Sci 880:263-80; and Reiter, Y. (1996) Clin Cancer Res 2:245-52). Single-chain antibodies can be dimerized or multimerized to produce polyvalent antibodies characterized to different epitopes of the same target protein.

[0248] In yet another embodiment, the antibody molecule has a heavy chain constant region selected from, for example, the heavy chain constant regions of IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE, particularly the (e.g., human) heavy chain constant region of, for example, IgG1, IgG2, IgG3, and IgG4. In another embodiment, the antibody molecule has a light chain constant region selected from, for example, the light chain constant region of kappa or lambda, for example, the (e.g., human) light chain constant region. The constant region can be modified, e.g., mutated, to modify the properties of the antibody (e.g., Fc receptor binding, antibody glycosylation, cysteine ​​residue count, increase or decrease of one or more effector cell functions and / or complement functions). In one embodiment, the antibody has effector function and can fix complement. In another embodiment, the antibody does not recruit effector cells or fix complement. In another embodiment, the antibody has reduced ability to bind to the Fc receptor or does not have such ability. For example, isotypes or subtypes, fragments or other variants that do not support binding to the Fc receptor, and which have, for example, a mutated or deficient Fc receptor binding region.

[0249] Methods for modifying the constant region of an antibody are known in this field. An antibody with modified function, such as a modified affinity for an effector ligand like FcR on cells or the C1 component of complement, can be produced by replacing at least one amino acid residue in the constant region of the antibody with a different residue (see, for example, EP388,151A1, U.S. Patent No. 5,624,821, and U.S. Patent No. 5,648,260 (all of which are incorporated herein by reference)). Similar types of modifications may also be described in which immunoglobulins would reduce these functions if applied to mice or other species.

[0250] Antibody molecules can be derivatized or conjugated to other functional molecules (e.g., other peptides or proteins). As used herein, “derivativeized” antibody molecules are modified. Methods of derivatization include, but are not limited to, the addition of affinity ligands such as fluorescent moieties, radionucleotides, toxins, enzymes, or biotin. Therefore, the antibody molecules of the present invention are intended to include modified forms other than those described herein, including derivatized and immunoadhesion molecules. For example, an antibody molecule can functionally conjugate (by chemical coupling, gene fusion, non-covalent bonding, or otherwise) to one or more other molecules, such as other antibodies (e.g., bispecific antibodies or bispecific antibodies), detectable drugs, cytotoxic agents, drugs, and / or other molecules, or proteins or peptides (e.g., streptavidin core regions or polyhistidine tags) that can mediate the binding of an antibody or antibody moiety to those molecules.

[0251] One type of derivatized antibody molecule is produced by crosslinking one or more antibodies (of the same type or, for example, different types to produce a bispecific antibody). Suitable crosslinkers include heterobifunctional (e.g., m-maleimidobenzoyl-N-hydroxysuccinimide) or homobifunctional (e.g., disaxinimidyl suberate) compounds having two distinctly reactive groups separated by a suitable spacer. Such linkers are available from Pierce Chemical Company, Rockford, Ill.

[0252] Useful detectable agents that can derivatize (or label) the antibody molecules of the present invention include fluorescent compounds, various enzymes, prosthetic groups, luminescent substances, bioluminescent substances, fluorescently emitting metal atoms such as europium (Eu) and other lanthanides, and radioactive materials. Representative fluorescent detectable agents include fluorescein, fluorescein isothiocyanate, rhodamine, 5-dimethylamine-1-naphthalenesulfonyl chloride, and phycoerythrin. Antibodies may also be derivatized with detectable enzymes such as alkaline phosphatase, horseradish peroxidase, β-galactosidase, acetylcholinesterase, and glucose oxidase. When antibodies are derivatized with detectable enzymes, they can be detected by adding further reagents that the enzyme uses to produce a detectable reaction product. For example, in the presence of the detectable agent horseradish peroxidase, the addition of hydrogen peroxide and diaminobenzidine produces a detectable colored reaction product. Antibody molecules may be derivatized with prosthetic groups (e.g., streptavidin / biotin and avidin / biotin). For example, an antibody can be derivatized with biotin and detected by indirect measurement of avidin or streptavidin binding. Suitable fluorescent substances include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansil chloride, or phycoerythrin; luminescent substances include luminol; and bioluminescent substances include luciferase, luciferin, and aequorin.

[0253] Labeled antibody molecules can be used in many situations, for example, diagnostically and / or in research, including (i) to isolate a target antibody by standard techniques such as affinity chromatography or immunoprecipitation; (ii) to detect a target antigen (e.g., in cell lysates or cell supernatants) to evaluate the quantity or expression pattern of a protein; and (iii) to monitor protein levels in tissue as part of a clinical trial method to determine the effectiveness of a treatment regimen.

[0254] Antibody molecules can be conjugated to other molecules, generally to label or therapeutic (e.g., cytotoxic or cell proliferation inhibitory) agents. Radioisotopes can be used in diagnostic or therapeutic applications. Radioisotopes that can bind to anti-PSMA antibodies include, but are not limited to, α-, β-, or γ-emitters or β- and γ-emitters. Such radioisotopes include iodine ( 131 I or 125 I), Yttrium ( 90 Y), Lutetium ( 177 Lu), Actinium ( 225 Ac), praseodymium, astatine ( 211 At), Rhenium ( 186 Re), bismuth ( 212 Bi or 213 Bi), Indium ( 111 In), technetium ( 99 mTc), phosphorus ( 32 P), Rhodium ( 188 Rh), sulfur ( 35 S), carbon ( 14 C), tritium ( 3 H), chromium ( 51 Cr), chlorine ( 36 Cl), cobalt ( 57 Co or 58 Co), iron ( 59 Fe), selenium ( 75 Se) or gallium ( 67 This includes, but is not limited to, Ga. Radioactive isotopes useful as therapeutic agents include yttrium ( 90 Y), Lutetium ( 177 Lu), Actinium ( 225 Ac), praseodymium, astatine ( 211 At), Rhenium ( 186 Re), bismuth ( 212 Bi or 213 Bi) and rhodium ( 188 It contains Rh. For example, radioactive isotopes useful as labels for use in diagnostic agents include iodine ( 131 I or 125 I), Indium ( 111 In), technetium (99 mTc), phosphorus ( 32 P), carbon ( 14 C) and tritium ( 3 H) or one or more of the therapeutic isotopes listed above.

[0255] This invention provides radiolabeled antibody molecules and methods for labeling them. In one embodiment, a method for labeling antibody molecules is disclosed. The method includes contacting an antibody molecule with a chelating agent to produce a conjugated antibody. The conjugated antibody is then labeled with a radioisotope, for example, 111 indium, 90 Yttrium and 177 Lutetium is used for labeling, thereby producing labeled antibody molecules.

[0256] As described above, antibody molecules can be conjugated with therapeutic agents. Therapeutic active radioisotopes have already been mentioned. Examples of other therapeutic agents include Taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, teniposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracinedione, mitoxantrone, mitramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, puromycin, mytansinoids, e.g., mytansinol (see U.S. Patent No. 5,208,020), CC-1065 (see U.S. Patents No. 5,475,092, 5,585,499, and 5,846,545) and their analogues or homologues. Therapeutic agents include, but are not limited to, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g., mechloretamine, thiotepa, chlorambucil, CC-1065, melphalan, carmustine (BSNU) and lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C and cis-dichlorodiamine platinum(II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mitramycin and anthramycin (AMC)), and antimitotic agents (e.g., vincristine, vinblastine, taxol and mytansinoids).

[0257] In one aspect, the present invention relates to a method for providing a target-binding molecule that specifically binds to the LAG-3 receptor. For example, the target-binding molecule is an antibody molecule. The method provides a target protein comprising at least a portion of a non-human protein, the portion being homologous to a corresponding portion of a human target protein (at least 70%, 75%, 80%, 85%, 87%, 90%, 92%, 94%, 95%, 96%, 97%, 98% identical), but differing by at least one amino acid (e.g., at least 1, 2, 3, 4, 5, 6, 7, 8, or 9 amino acids); obtaining an antibody molecule that specifically binds to an antigen; and evaluating the effect of the binder on modulating the activity of the target protein. The method may further include administering the binder (e.g., an antibody molecule) or a derivative (e.g., a humanized antibody molecule) to a human subject.

[0258] The present invention comprises an isolated nucleic acid molecule encoding the antibody molecule, its vector, and a host cell. The nucleic acid molecule includes, but is not limited to, RNA, genomic DNA, and cDNA.

[0259] In some embodiments, the antibody molecule is a multispecific (e.g., bispecific or triplicate) antibody molecule. Methods for producing bispecific or heterodimeric antibody molecules are known in the art, e.g., the “knob in hole” approach described in US5731168; e.g., electrostatic steering Fc pair formation described in WO09 / 089004, WO06 / 106905 and WO2010 / 129304; e.g., chain exchange domain (SEED) heterodimer formation described in WO07 / 110205; e.g., WO08 / 119353, WO2011 / 131746 and WO2013 / 060867 Fab arm replacement as described in the issue; for example, bi-antibody conjugates by antibody crosslinking to produce bistructural structures using heterobifunctional reactants having, for example, amine-reactive and sulfhydryl-reactive models, as described in US4433059; for example, bispecific antibody determinants produced by the recombination of hemiantibodies (heavy-light chain pairs or Fab) from different antibodies to reduce the reduction and oxidation cycles of the disulfide bond between two heavy chains, as described in US4444878; trifunctional antibodies, e.g., US527 For example, Fab' fragments of 3 crosslinked via sulfhydryl reactive groups, as described in No. 3743; biosynthetic binding proteins, for example, pairs of scFv crosslinked via the C-terminal tail, preferably by disulfide or amine reactive chemical crosslinking, as described in US No. 5534254; bifunctional antibodies, for example, Fab fragments with different specificities dimerized via leucine zippers (e.g., c-fos and c-jun) with substituted constant domains, as described in US No. 5582996; bispecificity and oligo-specific monovalent and oligovalent receptors, e.g., VH-CH1 regions of two antibodies (two Fab fragments) conjugated via a polypeptide spacer between the CH1 region of one antibody and the VH region of the other antibody with its generally accompanying light chain, as described in US5591828; bispecific DNA-antibody conjugates, e.g., crosslinking of antibodies or Fab fragments via double-strand mutation of DNA, as described in US5635602; bispecific fusion proteins, e.g., as described in US5637481Expression constructs comprising two scFvs, a hydrophilic helical peptide linker between them, and a fully constant region; polyvalent and polyspecific binding proteins, e.g., dimers of a first domain with a binding region for the Ig heavy chain variable region and a second domain with a binding region for the Ig light chain variable region, commonly called bispecific antibodies, as described in US5837242 (including higher-order structures that create bispecific, triplicate, or quadruplicate molecules); e.g., minibody constructs having bound VL and VH chains, further bound to the antibody hinge region and CH3 region by peptide spacers, which can be dimerized to form bispecific / polyvalent molecules, as described in US5837821; short peptide linkers of any orientation (e.g., 5 or 10) which can be dimerized to form bispecific antibodies. VH and VL domains linked with or without linkers having a mino acid; for example, trimers and tetramers as described in US5844094; for example, strings of VH domains (or family member VL domains) linked by peptide bonds to a C-terminal crosslinkable group, further linked to a VL domain to form a series of FVs (or scFvs) as described in US5864019; and single-chain linked polypeptides with both VH and VL domains linked by peptide linkers that combine into a multivalent structure by non-covalent or chemical crosslinking to form, for example, homobivalent, heterobivalent, trivalent, and tetravalent structures using both scFV or bispecific antibody type forms as described in US5869620. Further representative polyspecific and bispecific molecules and their manufacturing methods are, for example, US5910573, US5932448, US5959083, US5989830, US6005079, US6239259, US6294353, US6333396, US6476198, US6511663, US6670 No. 453, US6743896, US6809185, US6833441, US7129330, US7183076, US7521056, US752 No. 7787, No. US7534866, No. US7612181, No. US2002004587A1, No. US2002076406A1, No. US2002103345A1,US2003207346A1, US2003211078A1, US2004219643A1, US2004220388A1, US2004242847A1, US2005003403A1, US2005004352A1, US20 No. 05069552A1, No. US2005079170A1, No. US2005100543A1, No. US2005136049A1, No. US2005136051A1, No. US2005163782A1, No. US2005266425A1, No. US200608 No. 3747A1, No. US2006120960A1, No. US2006204493A1, No. US2006263367A1, No. US2007004909A1, No. US2007087381A1, No. US2007128150A1, No. US2007141049 A1, US2007154901A1, US2007274985A1, US2008050370A1, US2008069820A1, US2008152645A1, US2008171855A1, US2008241884A1, U S2008254512A1, US2008260738A1, US2009130106A1, US2009148905A1, US2009155275A1, US2009162359A1, US2009162360A1, US200 No. 9175851A1, No. US2009175867A1, No. US2009232811A1, No. US2009234105A1, No. US2009263392A1, No. US2009274649A1, No. EP346087A2, No. WO0006605A2, W This is seen in applications O02072635A2, WO04081051A1, WO06020258A2, WO2007044887A2, WO2007095338A2, WO2007137760A2, WO2008119353A1, WO2009021754A2, WO2009068630A1, WO9103493A1, WO9323537A1, WO9409131A1, WO9412625A2, WO9509917A1, WO9637621A2, and WO9964460A1. The contents of the above cited applications are incorporated herein by reference in their entirety. ,

[0260] In other embodiments, an anti-LAG-3 antibody molecule (e.g., a monospecific, bispecific, or polyspecific antibody molecule) is covalently bound, e.g., fused, to another partner, e.g., a protein, e.g., one, two, or more cytokines, e.g., as a fusion molecule, e.g., a fusion protein. In other embodiments, the fusion molecule comprises one or more proteins, e.g., one, two, or more cytokines. In one embodiment, the cytokine is an IL selected from one, two, three, or more interleukins (IL)-1, IL-2, IL-12, IL-15, or IL-21. In one embodiment, the bispecific antibody molecule has a first binding specificity to a first target (e.g., LAG-3) and a second binding specificity to a second target (e.g., PD-1, TIM-3, or PD-L1), and optionally binds to an interleukin (e.g., IL-12) domain, e.g., full-length IL-12 or a portion thereof.

[0261] A “fusion protein” and a “fusion polypeptide” are polypeptides having at least two covalently linked parts, where each part is a polypeptide with a different property. These properties can be biological properties, such as in vitro or in vivo activity. They can also be simple chemical or physical properties, such as binding to a target molecule or catalysis of a reaction. The two parts may be linked directly by a monopeptide bond or via a peptide linker, but they are within a leading frame.

[0262] The present invention provides isolated nucleic acid molecules encoding the antibody molecules described above, vectors thereof, and host cells. The nucleic acid molecules include, but are not limited to, RNA, genomic DNA, and cDNA.

[0263] Representative anti-LAG-3 antibody molecules In one embodiment, the anti-LAG-3 antibody molecule (i) the VHCDR1 amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 4, or SEQ ID NO: 286; the VHCDR2 amino acid sequence of SEQ ID NO: 2; and the heavy chain variable region (VH) including the VHCDR3 amino acid sequence of SEQ ID NO: 3; and (ii) The light chain variable region (VL) containing the VLCDR1 amino acid sequence of SEQ ID NO: 10, the VLCDR2 amino acid sequence of SEQ ID NO: 11, and the VLCDR3 amino acid sequence of SEQ ID NO: 12. Includes.

[0264] In other embodiments, the anti-LAG-3 antibody molecule is (i) VHCDR1 amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 4, or SEQ ID NO: 286; heavy chain variable region (VH) including the VHCDR2 amino acid sequence of SEQ ID NO: 5 and the VHCDR3 amino acid sequence of SEQ ID NO: 3; and (ii) The light chain variable region (VL) containing the VLCDR1 amino acid sequence of SEQ ID NO: 13, the VLCDR2 amino acid sequence of SEQ ID NO: 14, and the VLCDR3 amino acid sequence of SEQ ID NO: 15. Includes.

[0265] In one embodiment of the antibody molecule, VHCDR1 contains the amino acid sequence of SEQ ID NO: 1. In another embodiment, VHCDR1 contains the amino acid sequence of SEQ ID NO: 4. In yet another embodiment, VHCDR1 contains the amino acid sequence of SEQ ID NO: 286.

[0266] In one embodiment, the antibody molecule has a heavy chain variable region that includes at least one framework (FW) region containing an amino acid sequence that is at least 90% identical thereto to any of the amino acid sequences of SEQ ID NOs. 187, 190, 194, 196, 198, 202, 206, 208, 210, 212, 217, 219, or 221, or an amino acid sequence that has no more than two amino acid substitutions, insertions, or deletions compared to any of the amino acid sequences of SEQ ID NOs. 187, 190, 194, 196, 198, 202, 206, 208, 210, 212, 217, 219, or 221.

[0267] In another embodiment, the antibody molecule has a heavy chain variable region that includes at least one framework region containing any of the amino acid sequences of SEQ ID NOs: 187, 190, 194, 196, 198, 202, 206, 208, 210, 212, 217, 219, or 221.

[0268] In yet another embodiment, the antibody molecule has a heavy chain variable region comprising at least two, three, or four framework regions containing any of the amino acid sequences of SEQ ID NOs: 187, 190, 194, 196, 198, 202, 206, 208, 210, 212, 217, 219, or 221.

[0269] In another embodiment, the antibody molecule comprises the VHFW1 amino acid sequence of SEQ ID NO: 187, 190, 194, or 196, the VHFW2 amino acid sequence of SEQ ID NO: 198, 202, 206, or 208, and the VHFW3 amino acid sequence of SEQ ID NO: 210, 212, 217, or 219, and optionally further comprises the VHFW4 amino acid sequence of SEQ ID NO: 221.

[0270] In another embodiment, the antibody molecule has amino acids having variable light chain substitutions, insertions, or deletions, including at least one framework region containing an amino acid sequence that is at least 90% identical thereto to any of the amino acid sequences of SEQ ID NOs: 226, 230, 232, 234, 236, 238, 240, 244, 246, 248, 252, 255, 259, 261, 265, 267, 269, or 271, or an amino acid sequence having no more than 2 amino acid substitutions, insertions, or deletions compared to any of the amino acid sequences of SEQ ID NOs: 226, 230, 232, 234, 236, 238, 240, 244, 246, 248, 252, 255, 259, 261, 265, 267, 269, or 271.

[0271] In another embodiment, the antibody molecule has a light chain variable region that includes at least one framework region containing any of the amino acid sequences of light chain variable region SEQ ID NOs: 226, 230, 232, 234, 236, 238, 240, 244, 246, 248, 252, 255, 259, 261, 265, 267, 269, or 271.

[0272] In another embodiment, the antibody molecule has a light chain variable region comprising at least two, three, or four framework regions containing any of the amino acid sequences of SEQ ID NOs: 226, 230, 232, 234, 236, 238, 240, 244, 246, 248, 252, 255, 259, 261, 265, 267, 269, or 271.

[0273] In another embodiment, the antibody molecule comprises the VLFW1 amino acid sequence of SEQ ID NO: 226, 230, 232, 234, 236, or 238, the VLFW2 amino acid sequence of SEQ ID NO: 240, 244, 246, or 248, and the VLFW3 amino acid sequence of SEQ ID NO: 252, 255, 259, 261, 265, 267, or 269, and optionally further comprises the VLFW4 amino acid sequence of SEQ ID NO: 271.

[0274] In another embodiment, the antibody molecule includes a heavy chain variable domain having an amino acid sequence that is at least 85% identical to any of SEQ ID NOs: 8, 28, 64, 68, 72, 76, 80, 100, 104, or 108.

[0275] In another embodiment, the antibody molecule includes a heavy chain variable domain comprising the amino acid sequence of SEQ ID NOs: 8, 28, 64, 68, 72, 76, 80, 100, 104, or 108.

[0276] In another embodiment, the antibody molecule includes a light chain variable domain having an amino acid sequence that is at least 85% identical to any of SEQ ID NOs: 32, 36, 40, 44, 48, 52, 56, 60, 84, 88, 92, or 96.

[0277] In another embodiment, the antibody molecule includes a light chain variable domain comprising the amino acid sequence of SEQ ID NOs: 32, 36, 40, 44, 48, 52, 56, 60, 84, 88, 92, or 96.

[0278] In another embodiment, the antibody molecule includes a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 8.

[0279] In another embodiment, the antibody molecule comprises a heavy chain having the amino acid sequence of SEQ ID NO: 18.

[0280] In another embodiment, the antibody molecule includes a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 28.

[0281] In another embodiment, the antibody molecule comprises a heavy chain having the amino acid sequence of SEQ ID NO: 30.

[0282] In another embodiment, the antibody molecule includes a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 64.

[0283] In another embodiment, the antibody molecule comprises a heavy chain having the amino acid sequence of SEQ ID NO: 66.

[0284] In another embodiment, the antibody molecule includes a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 68.

[0285] In another embodiment, the antibody molecule comprises a heavy chain having the amino acid sequence of SEQ ID NO: 70.

[0286] In another embodiment, the antibody molecule includes a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 72.

[0287] In another embodiment, the antibody molecule comprises a heavy chain having the amino acid sequence of SEQ ID NO: 74.

[0288] In another embodiment, the antibody molecule includes a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 76.

[0289] In another embodiment, the antibody molecule comprises a heavy chain having the amino acid sequence of SEQ ID NO: 78.

[0290] In another embodiment, the antibody molecule includes a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 80.

[0291] In another embodiment, the antibody molecule comprises a heavy chain having the amino acid sequence of SEQ ID NO: 82.

[0292] In another embodiment, the antibody molecule includes a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 100.

[0293] In another embodiment, the antibody molecule comprises a heavy chain having the amino acid sequence of SEQ ID NO: 102 or SEQ ID NO: 113.

[0294] In another embodiment, the antibody molecule includes a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 104.

[0295] In another embodiment, the antibody molecule comprises a heavy chain having the amino acid sequence of SEQ ID NO: 106.

[0296] In another embodiment, the antibody molecule comprises a heavy chain having the amino acid sequence of SEQ ID NO: 122.

[0297] In another embodiment, the antibody molecule includes a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 108.

[0298] In another embodiment, the antibody molecule comprises a heavy chain having the amino acid sequence of SEQ ID NO: 110.

[0299] In another embodiment, the antibody molecule comprises a heavy chain having the amino acid sequence of SEQ ID NO: 134.

[0300] In another embodiment, the antibody molecule includes a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 32.

[0301] In another embodiment, the antibody molecule comprises a light chain having the amino acid sequence of SEQ ID NO: 34.

[0302] In another embodiment, the antibody molecule includes a light chain variable domain containing the amino acid sequence of SEQ ID NO: 36.

[0303] In another embodiment, the antibody molecule includes a light chain having the amino acid sequence of SEQ ID NO: 38.

[0304] In another embodiment, the antibody molecule includes a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 40.

[0305] In another embodiment, the antibody molecule includes a light chain having the amino acid sequence of SEQ ID NO: 42.

[0306] In another embodiment, the antibody molecule includes a light chain variable domain containing the amino acid sequence of SEQ ID NO: 44.

[0307] In another embodiment, the antibody molecule includes a light chain having the amino acid sequence of SEQ ID NO: 46.

[0308] In another embodiment, the antibody molecule includes a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 48.

[0309] In another embodiment, the antibody molecule includes a light chain having the amino acid sequence of SEQ ID NO: 50.

[0310] In another embodiment, the antibody molecule includes a light chain variable domain containing the amino acid sequence of SEQ ID NO: 52.

[0311] In another embodiment, the antibody molecule includes a light chain having the amino acid sequence of SEQ ID NO: 54.

[0312] In another embodiment, the antibody molecule includes a light chain variable domain containing the amino acid sequence of SEQ ID NO: 56.

[0313] In another embodiment, the antibody molecule includes a light chain having the amino acid sequence of SEQ ID NO: 58.

[0314] In another embodiment, the antibody molecule includes a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 60.

[0315] In another embodiment, the antibody molecule includes a light chain having the amino acid sequence of SEQ ID NO: 62.

[0316] In another embodiment, the antibody molecule includes a light chain variable domain containing the amino acid sequence of SEQ ID NO: 84.

[0317] In another embodiment, the antibody molecule includes a light chain having the amino acid sequence of SEQ ID NO: 86.

[0318] In another embodiment, the antibody molecule includes a light chain variable domain containing the amino acid sequence of SEQ ID NO: 88.

[0319] In another embodiment, the antibody molecule includes a light chain having the amino acid sequence of SEQ ID NO: 90.

[0320] In another embodiment, the antibody molecule includes a light chain variable domain containing the amino acid sequence of SEQ ID NO: 92.

[0321] In another embodiment, the antibody molecule includes a light chain having the amino acid sequence of SEQ ID NO: 94.

[0322] In another embodiment, the antibody molecule includes a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 96.

[0323] In another embodiment, the antibody molecule includes a light chain having the amino acid sequence of SEQ ID NO: 98.

[0324] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 32.

[0325] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 36.

[0326] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 40.

[0327] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 44.

[0328] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 48.

[0329] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 52.

[0330] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 56.

[0331] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 60.

[0332] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 64 or SEQ ID NO: 104; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 36.

[0333] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 64 or SEQ ID NO: 104; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 40.

[0334] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 64 or SEQ ID NO: 104; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 56.

[0335] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 64 or SEQ ID NO: 104; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 60.

[0336] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 68 or SEQ ID NO: 108; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 36.

[0337] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 72 or SEQ ID NO: 8; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 40.

[0338] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 72 or SEQ ID NO: 8; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 60.

[0339] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 76 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 60.

[0340] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 80 and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 84.

[0341] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 88.

[0342] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 92.

[0343] In another embodiment, the antibody molecule comprises a heavy chain variable domain containing the amino acid sequence of SEQ ID NO: 64 or SEQ ID NO: 104; and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 96.

[0344] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 34.

[0345] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 38.

[0346] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 42.

[0347] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 46.

[0348] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 50.

[0349] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 54.

[0350] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 58.

[0351] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 62.

[0352] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 66 or SEQ ID NO: 106; and a light chain containing the amino acid sequence of SEQ ID NO: 38.

[0353] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 66 or SEQ ID NO: 106; and a light chain containing the amino acid sequence of SEQ ID NO: 42.

[0354] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 66 or SEQ ID NO: 106; and a light chain containing the amino acid sequence of SEQ ID NO: 58.

[0355] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 66 or SEQ ID NO: 106; and a light chain containing the amino acid sequence of SEQ ID NO: 62.

[0356] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 70 or SEQ ID NO: 110; and a light chain containing the amino acid sequence of SEQ ID NO: 38.

[0357] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 74 or SEQ ID NO: 18; and a light chain containing the amino acid sequence of SEQ ID NO: 42.

[0358] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 74 or SEQ ID NO: 18; and a light chain containing the amino acid sequence of SEQ ID NO: 62.

[0359] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 78 and a light chain containing the amino acid sequence of SEQ ID NO: 62.

[0360] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 82 and a light chain containing the amino acid sequence of SEQ ID NO: 86.

[0361] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 94.

[0362] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 66 or SEQ ID NO: 106; and a light chain containing the amino acid sequence of SEQ ID NO: 98.

[0363] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 113 and a light chain containing the amino acid sequence of SEQ ID NO: 34.

[0364] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 113 and a light chain containing the amino acid sequence of SEQ ID NO: 38.

[0365] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 122 and a light chain containing the amino acid sequence of SEQ ID NO: 38.

[0366] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 122 and a light chain containing the amino acid sequence of SEQ ID NO: 58.

[0367] In another embodiment, the antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 134 and a light chain containing the amino acid sequence of SEQ ID NO: 38.

[0368] In other embodiments, the antibody molecule is selected from Fab, F(ab')2, Fv, or a single-stranded Fv fragment (scFv).

[0369] In another embodiment, the antibody molecule comprises a heavy chain constant region selected from IgG1, IgG2, IgG3, and IgG4.

[0370] In another embodiment, the antibody molecule includes a light chain constant region selected from the kappa or lambda light chain constant regions.

[0371] In another embodiment, the antibody molecule comprises a human IgG quadruplex constant region and a kappa light chain constant region having a mutation at position 228 according to EU numbering or at position 108 of SEQ ID NO: 275 or 277.

[0372] In another embodiment, the antibody molecule comprises a human IgG 4-chain constant region and a kappa light chain constant region having a serine-to-proline mutation at position 228 according to EU numbering or at position 108 of SEQ ID NO: 275 or 277.

[0373] In another embodiment, the antibody molecule comprises a human IgG1 heavy chain constant region and a kappa light chain constant region having an asparagine-to-alanine mutation at position 297 according to EU numbering or position 180 of SEQ ID NO: 279.

[0374] In another embodiment, the antibody molecule comprises a human IgG1 heavy chain constant region and a kappa light chain constant region having an aspartic acid to alanine mutation at EU numbering position 265 or 148 and a proline to alanine mutation at EU numbering position 329 or position 212 of SEQ ID NO: 280.

[0375] In another embodiment, the antibody molecule comprises a human IgG1 heavy chain constant region and a kappa light chain constant region having a leucine-to-alanine mutation at EU numbering position 234 or 117 and a leucine-to-alanine mutation at EU numbering position 235 or position 118 of SEQ ID NO: 281.

[0376] In another embodiment, the antibody molecule has a dissociation constant (K) less than about 0.2 nM. D It can bind to human LAG-3.

[0377] In certain embodiments, the antibody molecule binds to human LAG-3 with a K, for example, measured by the Biacore method, of less than about 0.2 nM, 0.15 nM, 0.1 nM, 0.05 nM or 0.02 nM, such as about 0.05 nM to 0.15 nM, such as about 0.11 nM D

[0378] In other embodiments, the antibody molecule binds to cynomolgus LAG-3 with a K, for example, measured by the Biacore method, of less than about 0.2 nM, 0.15 nM, 0.1 nM, 0.05 nM or 0.02 nM, such as about 0.05 nM to 0.15 nM D

[0379] In certain embodiments, the antibody molecule binds with a K, for example, measured by the Biacore method, that is similar for both human LAG-3 and cynomolgus LAG-3, such as in the nM range D D In certain embodiments, the antibody molecule binds to the human LAG-3-Ig fusion protein with a K, for example, measured by ELISA, of less than about 0.5 nM, 0.2 nM, 0.1 nM, 0.05 nM, 0.025 nM or 0.01 nM

[0380] In certain embodiments, the antibody molecule binds to CHO cells expressing human LAG-3 (e.g., human LAG-3 transfected CHO cells) with a K, for example, measured by FACS analysis, of less than about 4 nM, 2.5 nM, 2 nM, 1.5 nM, 1 nM, 0.75 nM, 0.5 nM, 0.4 nM, 0.3 nM, 0.2 nM, 0.1 nM or 0.05 nM, such as about 2.3 nM, 1.92 nM or about 0.2 nM D

[0381] In certain embodiments, the antibody molecule binds to human T cells with a K, for example, measured by FACS analysis, of less than about 0.5 nM, 0.4 nM, 0.3 nM, 0.2 nM, 0.1 nM or 0.05 nM, such as about 0.26 nM D

[0382] ​​​​​In one embodiment, the antibody molecule is measured, for example by FACS analysis, to have a K content of approximately 20 nM, 15 nM, 10 nM, 5 nM, 2 nM, or less than 1 nM in cells expressing LAG-3 (e.g., human LAG-3 expressing 300.19 cells), for example, approximately 13.6 nM. D They are joined together.

[0383] In one embodiment, the antibody molecule is measured, for example by FACS analysis, to have a K content of approximately 15 nM, 10 nM, 9 nM, 8 nM, 6 nM, 5 nM, 2 nM, or less than 1 nM in cells expressing rhesus monkey LAG-3 (e.g., cells transfected with rhesus monkey LAG-3), for example, approximately 8.03 nM. D They are joined together.

[0384] In one embodiment, the antibody molecule is not cross-reactive with mouse LAG-3. In another embodiment, the antibody is not cross-reactive with rat LAG-3. In yet another embodiment, the antibody is cross-reactive with rhesus monkey LAG-3. In yet another embodiment, the antibody is cross-reactive with rat LAG-3. For example, cross-reactivity can be measured by the Biacore method or a binding assay using cells expressing LAG-3 (e.g., human LAG-3 expressing 300.19 cells).

[0385] In other embodiments, the antibody molecule binds to any of the extracellular Ig-like domains of LAG-3 (e.g., human LAG-3), e.g., domain 1 (D1), domain 2 (D2), domain 3 (D3), or domain 4 (D4). In some embodiments, the antibody molecule binds to one or more amino acid residues in D1. In some embodiments, the antibody molecule does not bind to the extra loop or fragment of D1 (e.g., as measured by Biacore or FACS). In some embodiments, the antibody does not bind to D2. In some embodiments, the antibody molecule binds to both D1 and D2. In some embodiments, the antibody molecule binds to one or more amino acid residues in D1 and / or D2 that bind to MHC class II molecules. In other embodiments, the antibody molecule can reduce the binding of LAG-3 to major histocompatibility (MHC) class II molecules or cells expressing MHC class II molecules. In one embodiment, the antibody molecule has an IC50 concentration of about 10 nM, 8 nM, 5 nM, 4 nM, 3 nM, 2 nM, 1 nM, or less than 0.5 nM, for example, about 8 nM to about 10 nM or about 2 nM to about 3 nM, for example, about 5.5 nM or about 2.3 nM. 50 For example, this reduces (e.g., blocks) LAG-3-Ig binding to MHC class II molecules in Raji cells or Daudi cells.

[0386] In another aspect, the antibody molecule can enhance the antigen-specific T cell response.

[0387] In some embodiments, the antibody molecule is a monospecific antibody molecule or a bispecific antibody molecule. In some embodiments, the antibody molecule has first binding specificity to LAG-3 and second binding specificity to PD-1, TIM-3, CEACAM (e.g., CEACAM-1 and / or CEACAM-5), PD-L1, or PD-L2. In some embodiments, the antibody molecule has an antigen-binding fragment of the antibody, e.g., a half-antibody or an antigen-binding fragment of a half-antibody.

[0388] In one embodiment, the antibody molecule increases IL-2 expression from cells activated with Staphylococcus enterotoxin B (SEB) (e.g., 25 μg / mL) by at least approximately 2-3 times, 3-4-5 times, for example, approximately 2-3 times, compared to IL-2 expression using an isotype control (e.g., IgG4), as measured, for example, in an SEB T cell activation assay or a human whole blood ex vivo assay.

[0389] In one embodiment, the antibody molecule increases IFN-γ expression from T cells stimulated with anti-CD3 (e.g., 0.1 μg / mL) by at least about 0.5, 1, 2, 3, 4, 5, 6, 7, or 8 times, for example about 0.9 to 5.1 times, for example about 3 times, compared to IFN-γ expression using an isotype control (e.g., IgG4), as measured, for example, in an IFN-γ activity assay.

[0390] In one embodiment, the antibody molecule increases IFN-γ expression from T cells activated by SEB (e.g., 3 pg / mL) by at least approximately 2-fold, 3-fold, 4-fold, 5-fold, for example, approximately 1.2-2-fold, for example, approximately 1.6-fold, compared to IFN-γ expression using an isotype control (e.g., IgG4), as measured, for example, in an IFN-γ activity assay.

[0391] In one embodiment, the antibody molecule does not increase the expression of IL-2 or IFN-γ without T cell receptor activation (e.g., in the absence of SEB).

[0392] In one embodiment, the antibody molecule increases IFN-γ expression from CMV peptide-activated T cells by at least approximately 2-fold, 3-fold, 4-fold, 5-fold, for example, approximately 1.1-1.7-fold, for example, approximately 1.4-fold, compared to IFN-γ expression using an isotype control (e.g., IgG4), as measured, for example, in an IFN-γ activity assay. In one embodiment, the antibody molecule is CMV peptide-activated CD8 + T cell proliferation is, for example, CD8 cells that have gone through at least n (e.g., n=2 or 4) cell divisions. +Measured by the percentage of T cells, CD8 when using an isotype control (e.g., IgG4) + T cell proliferation is increased by at least about 1, 2-fold, 3-fold, 4-fold, 5-fold, e.g., about 1.5-fold compared to that of the isotype control (e.g., IgG4).

[0393] In certain embodiments, the antibody molecule has a C of about 50 μg / mL to about 400 μg / mL, about 100 μg / mL to about 350 μg / mL, about 150 μg / mL to about 300 μg / mL or about 200 μg / mL to about 250 μg / mL, e.g., about 166 μg / mL, measured in an animal, for example max has.

[0394] In certain embodiments, the antibody molecule has a T of about 50 hours to about 400 hours, about 100 hours to about 350 hours, about 150 hours to about 300 hours or about 200 hours to about 250 hours, e.g., about 231.9 hours, measured in an animal, for example 1 / 2 [[ID=​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​In other respects, the present invention provides isolated nucleic acid molecules encoding any of the aforementioned antibody molecules, vectors thereof, and host cells.

[0397] In one embodiment, the isolated nucleic acid encodes either the antibody heavy chain variable region or the light chain variable region or both of the antibody molecule.

[0398] In one embodiment, the isolated nucleic acid encodes a heavy chain CDR1-3, where the nucleic acid comprises the nucleotide sequences of SEQ ID NOs. 140-144, 151-155, 162-166, 173-177, 184-186, or 287.

[0399] In another embodiment, the isolated nucleic acid encodes light chain CDR1-3, where the nucleic acid comprises the nucleotide sequences of SEQ ID NOs. 145-150, 156-161, 167-172, or 178-183.

[0400] In another embodiment, the nucleic acid further comprises a nucleotide sequence encoding a heavy chain variable domain, wherein the nucleotide sequence is at least 85% identical to any of SEQ ID NOs: 9, 29, 65, 69, 73, 77, 81, 101, 105, 109, 112, 121, 124, 125, 132, or 133.

[0401] In another embodiment, the nucleic acid further comprises a nucleotide sequence encoding a heavy chain variable domain, wherein the nucleotide sequence includes any of SEQ ID NOs: 9, 29, 65, 69, 73, 77, 81, 101, 105, 109, 112, 121, 124, 125, 132, or 133.

[0402] In another embodiment, the nucleic acid further comprises a nucleotide sequence encoding a heavy chain, wherein the nucleotide sequence is at least 85% identical to any of SEQ ID NOs: 19, 31, 67, 71, 75, 79, 83, 103, 107, 111, 114, 123, 126, 127, 135, or 136.

[0403] In another embodiment, the nucleic acid further comprises a nucleotide sequence encoding a heavy chain, wherein the nucleotide sequence includes any of SEQ ID NOs: 19, 31, 67, 71, 75, 79, 83, 103, 107, 111, 114, 123, 126, 127, 135, or 136.

[0404] In another embodiment, the nucleic acid further comprises a nucleotide sequence encoding a light chain variable domain, wherein the nucleotide sequence is at least 85% identical to any of SEQ ID NOs: 33, 37, 41, 45, 49, 53, 57, 61, 85, 89, 93, 97, 115, 118, 128, 129, or 137.

[0405] In another embodiment, the nucleic acid further comprises a nucleotide sequence encoding a light chain variable domain, wherein the nucleotide sequence includes any of SEQ ID NOs: 33, 37, 41, 45, 49, 53, 57, 61, 85, 89, 93, 97, 115, 118, 128, 129, or 137.

[0406] In another embodiment, the nucleic acid further comprises a nucleotide sequence encoding a light chain, wherein the nucleotide sequence is at least 85% identical to any of SEQ ID NOs: 35, 39, 43, 47, 51, 55, 59, 63, 87, 91, 95, 99, 117, 120, 130, 131, 138, or 139.

[0407] In another embodiment, the nucleic acid further comprises a nucleotide sequence encoding a light chain, wherein the nucleotide sequence includes any of SEQ ID NOs: 35, 39, 43, 47, 51, 55, 59, 63, 87, 91, 95, 99, 117, 120, 130, 131, 138, or 139.

[0408] In one embodiment, one or more expression vectors containing the nucleic acid and host cells are provided.

[0409] We also provide a method for producing antibody molecules or fragments thereof, which includes culturing host cells as described herein under conditions suitable for gene expression.

[0410] Pharmaceutical compositions and kits In other words, the present invention provides compositions comprising the antibody molecules described herein, formulated with a pharmaceutically acceptable carrier, for example, a pharmaceutically acceptable composition. The “pharmaceutically acceptable carrier” as used herein includes any and all physiologically compatible solvents, dispersion media, isotonic agents, and absorption retarders. The carrier may be suitable for intravenous, intramuscular, subcutaneous, non-enteral, rectal, spinal, or epithelial administration (e.g., by injection or infusion).

[0411] The compositions of the present invention may take various forms. These include liquid, semi-solid, and solid dosage forms, such as liquid solutions (e.g., injectable and drip-proof solutions), dispersants or suspensions, liposomes, and suppositories. The preferred form depends on the intended method of administration and therapeutic application. Generally, the preferred composition is in the form of an injectable or drip-proof solution. Preferred methods of administration are non-enteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). In a preferred embodiment, the antibody is administered by intravenous drip or injection. In another preferred embodiment, the antibody is administered by intramuscular or subcutaneous injection.

[0412] The terms “non-enteral administration” and “administered non-enterally” as used herein refer to methods of administration other than enteral and local administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intra-arterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions.

[0413] Therapeutic compositions must be sterile and stable under manufacturing and storage conditions. Compositions can be formulated as solutions, microemulsions, dispersants, liposomes, or other indicated configurations suitable for high antibody concentrations. Sterile injectable solutions can be prepared by incorporating the required amount of the active compound (i.e., antibody or antibody moiety) into a suitable solvent containing one or a combination of the components listed above, and then, if necessary, by filtration sterilization. Generally, dispersants are prepared by incorporating the active compound into a sterile medium containing a basic dispersion medium and the necessary components from those listed above. For sterile powders for the preparation of sterile injectable solutions, preferred methods of preparation are vacuum drying and lyophilization, which produce a powder of the active component and any further desired components from a pre-filtered solution. Appropriate fluidity of the solution can be maintained, for example, by the use of a coating such as lecithin, by maintaining the required particle size in the case of dispersants, and by the use of surfactants. Long-term absorption of injectable compositions can be achieved by incorporating absorption-delaying agents, such as monostearate and gelatin, into the composition.

[0414] Antibody molecules can be administered by a variety of methods known in this field, but for many therapeutic applications, the preferred route / method of administration is intravenous injection or infusion. In one embodiment, the antibody molecule is administered at approximately 35-440 mg / m². 2 Preferably about 70-310 mg / m² 2 Comfortably, approximately 110-130 mg / m² 2 To reach the dose, the antibody molecule is administered by intravenous infusion at a rate faster than 20 mg / min, for example, 20-40 mg / min, preferably 40 mg / min or higher. In another embodiment, the antibody molecule is administered at a rate of approximately 1-100 mg / m³. 2 Preferably about 5-50 mg / m² 2 , about 7~25mg / m 2 Comfortably, approximately 10 mg / m² 2To reach the desired dose, the drug is administered by intravenous infusion at a rate of less than 10 mg / min, preferably 5 mg / min or less. As will be recognized by those skilled in the art, the route / method of administration varies depending on the intended outcome. In some embodiments, the active compound may be formulated with a carrier that protects the compound drug from rapid release, such as controlled-release formulations, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydride, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. Many methods for producing such formulations are patented or generally known to those skilled in the art. See, for example, Sustained and Controlled Release Drug Delivery Systems, JR Robinson, ed., Marcel Dekker, Inc., New York, 1978.

[0415] In one embodiment, antibody molecules can be administered orally, for example, with an inert diluent or an absorbable edible carrier. The compound (and, if desired, other components) may also be filled into hard or soft-shelled gelatin, compressed into tablets, or directly incorporated into the target diet. For oral therapeutic administration, the compound can be incorporated with additives and may be used in the form of ingestible tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, wafers, etc. To administer the compound of the present invention by means other than enteral administration, it may be necessary to coat the compound with a substance that inhibits its inactivation, or to co-administer the compound with such a substance. Therapeutic compositions can also be administered using medical devices known in the art.

[0416] The administration regimen is adjusted to provide the optimal desired response (e.g., a therapeutic response). For example, a single bolus may be administered, divided doses may be administered over time, or the dose may be proportionally decreased or increased, as indicated by the presence of the treatment situation. For ease of administration and dose uniformity, it is particularly advantageous to formulate non-enteral compositions in dose-unit forms. The dose-unit forms used herein refer to physically separate units suitable as unit doses for the subject to be treated, each unit containing a predetermined amount of the active compound calculated to produce the desired therapeutic effect, along with the necessary pharmaceutical carrier. The details of the dose-unit forms of the present invention are specified and directly depend on (a) the unique characteristics of the active compound and the specific therapeutic effect to be achieved, and (b) the limitations inherent in the field in the manufacture of such active compounds for the sensitivity of the treatment in the individual.

[0417] A typical, non-limiting range for the therapeutically or prophylactically effective dose of the antibody molecule is 0.1–30 mg / kg, more preferably 1–25 mg / kg. The dosage and treatment regimen of the anti-LAG-3 antibody molecule can be determined by those skilled in the art. In one embodiment, the anti-LAG-3 antibody molecule is administered by injection (e.g., subcutaneously or intravenously) at doses of about 1–40 mg / kg, e.g., 1–30 mg / kg, e.g., about 5–25 mg / kg, about 10–20 mg / kg, about 1–5 mg / kg, 1–10 mg / kg, 5–15 mg / kg, 10–20 mg / kg, 15–25 mg / kg, or about 3 mg / kg. The dosing schedule can vary, for example, from once a week to once every two, three, or four weeks. In one embodiment, the anti-LAG-3 antibody molecule is administered every other week at a dose of about 10–20 mg / kg. Antibody molecules, approximately 35-440 mg / m² 2 Preferably about 70-310 mg / m² 2 Comfortably, approximately 110-130 mg / m² 2 To reach the desired dose, the drug is administered by intravenous infusion at a rate faster than 20 mg / min, for example, 20-40 mg / min, preferably 40 mg / min or higher. In one embodiment, approximately 110-130 mg / min 2The infusion rate achieves a level of approximately 3 mg / kg. In one embodiment, the anti-LAG-3 antibody molecule is administered (e.g., intravenously) in doses of approximately 3 to 800 mg, for example, approximately 3 mg, 20 mg, 80 mg, 240 mg, or 800 mg. In one embodiment, the anti-LAG-3 antibody molecule is administered alone in doses of approximately 20 to 800 mg, for example, approximately 3 mg, 20 mg, 80 mg, 240 mg, or 800 mg. In another embodiment, the anti-LAG-3 antibody molecule is administered in doses of approximately 3 to 240 mg, for example, approximately 3 mg, 20 mg, 80 mg, or 240 mg, in combination with a second agent or therapeutic modality, for example, a second agent or therapeutic modality described herein. In one embodiment, the anti-LAG-3 antibody molecule is administered every two weeks (e.g., between weeks 1, 3, 5, and 7) during each 8-week cycle up to 96 weeks.

[0418] Antibody molecules, approximately 35-440 mg / m² 2 Preferably about 70-310 mg / m² 2 Comfortably, approximately 110-130 mg / m² 2 To reach the desired dose, it can be administered by intravenous infusion at a rate faster than 20 mg / min, for example, 20-40 mg / min, preferably 40 mg / min or higher. In one embodiment, approximately 110-130 mg / min 2 The injection rate achieves a level of approximately 3 mg / kg. In another embodiment, the antibody molecule is injected at approximately 1-100 mg / m³. 2 For example, approximately 5-50 mg / m² 2 , about 7~25mg / m 2 Comfortably, approximately 10 mg / m² 2 To reach the desired dose, the antibody is administered by intravenous infusion at a rate of less than 10 mg / min, for example, 5 mg / min or less. In one embodiment, the antibody is infused over a period of approximately 30 minutes.

[0419] It should be noted that dosage values ​​will vary depending on the type and severity of the condition to be alleviated. For any specific subject, a specific dosing regimen should be adjusted over time according to individual needs and at the professional discretion of the person administering or supervising the administration of the composition. It should be further understood that the dosage ranges shown herein are merely representative examples and are not intended to limit the scope or implementation of the claimed composition.

[0420] The pharmaceutical compositions of the present invention may contain a “therapeutic effective dose” or “preventive effective dose” of the antibody or antibody moiety of the present invention. “Therapeutic effective dose” refers to the amount and duration of time that is effective in a dose to achieve a desired therapeutic outcome. The therapeutic effective dose of a modified antibody or antibody fragment may vary depending on factors such as the individual’s disease state, age, sex, and weight, and the ability of the antibody or antibody moiety to produce the desired effect in the individual. The therapeutic effective dose also refers to the amount at which the therapeutic effect outweighs any toxic or adverse effects of the modified antibody or antibody fragment. A “therapeutic effective dose” preferably inhibits a measurable parameter, such as tumor growth rate, by at least about 20%, more preferably at least about 40%, even less preferably about 60%, and still more preferably at least about 80% in an untreated subject. Measurable parameters of a compound, such as its ability to inhibit cancer, can be evaluated in animal model systems that predict efficacy in human tumors. Alternatively, the properties of the composition can be evaluated by testing the inhibitory ability of the compound, such as in vitro inhibition by assays known to those skilled in the art.

[0421] The “protective dose” refers to the amount and duration of the dose that is effective in achieving the desired protective effect. Generally, the protective dose is less than the therapeutic dose because the protective dose is used before or in the early stages of the disease.

[0422] Also within the scope of the present invention are kits comprising the antibody molecules described herein. The kit may comprise one or more other elements, including instructions for use; other agents, such as labels, therapeutic agents, or agents or radioprotective compositions useful for labeling or chelating or otherwise coupling antibodies with therapeutic agents; devices or other tools for preparing antibodies for administration; pharmaceutically acceptable carriers; and devices or other tools for administration to a subject.

[0423] Using anti-LAG-3 antibody molecule The anti-LAG-3 antibody molecules disclosed herein have diagnostic, therapeutic, and prophylactic utility in vitro and in vivo. For example, these molecules can enhance immunity by administering them to cells in culture in vitro or ex vivo, or to a subject, e.g., a human subject, in vivo. In one embodiment, the anti-LAG-3 antibody molecule enhances the immune response in a subject, for example, by blocking LAG-3 (e.g., by blocking LAG-3 binding to MHC molecules or other ligands).

[0424] Accordingly, in some respects, the present invention provides a method for modifying the immune response in a subject, comprising administering the subject an antibody molecule described herein so that the immune response in the subject is modified. In some embodiments, the immune response is enhanced, stimulated, or upregulated. In some embodiments, the anti-LAG-3 antibody molecule restores, enhances, or stimulates the antigen-specific T cell response in the subject, for example, the production of interleukin-2 (IL-2) or interferon-gamma (IFN-γ). In some embodiments, the immune response is an antitumor response. The methods and compositions described herein are suitable for the treatment of human patients having disorders that can be treated by enhancing the T cell-mediated immune response. For example, anti-LAG-3 antibody molecules, alone or in combination, can be administered to a subject to treat, prevent, and / or diagnose a variety of disorders, such as cancer (melanoma or liver cancer) or infectious disorders.

[0425] The term “subject” as used herein is intended to include humans and non-human animals. In some embodiments, the subject is a human subject, e.g., a human patient having a disorder or condition characterized by abnormal LAG-3 function. The term “non-human animal” in this invention includes mammals and non-mammals, e.g., non-human primates. In some embodiments, the subject is human. In some embodiments, the subject is a human patient requiring an enhanced immune response. In some embodiments, the subject has or is at risk of having one of the disorders described herein, e.g., cancer or infectious disorders described herein. In some embodiments, the subject is immunocompromised or at risk of becoming immunocompromised. For example, the subject is receiving or has received chemotherapy and / or radiotherapy. Separately or in combination therewith, the subject is immunocompromised or at risk of becoming immunocompromised as a result of infection. For example, the methods and compositions described herein enhance a number of immune activities. In some embodiments, the subject has an increased number or activity of tumor-infiltrating T lymphocytes (TILs). In other embodiments, the subject has an increased expression or activity of interferon-gamma (IFN-γ). In yet another embodiment, the subject has increased PD-L1 expression or activity. Therefore, in one embodiment, any of TIL, IFN-γ, CD8, or PD-L1 (e.g., 1, 2, 3, or all of them) can be used as biomarkers for the anti-LAG-3 immunotherapy described herein.

[0426] therapeutic use cancer Antibody-mediated blockade of LAG-3 can enhance the immune response against cancer cells in a target. Similar to CD4, LAG-3 interacts with MHC class II molecules, but unlike CD4, LAG-3 does not interact with the human immunodeficiency virus gp120 protein (Baixeras et al. (1992) J. Exp. Med. 176:327-337). Direct and specific binding of LAG-3 to MHC class II on the cell surface has been demonstrated in studies (Huard et al. (1996) Eur. J. Immunol. 26:1180-1186). The LAG-3 / MHC class II interaction is similar to that of CD4.+ and CD8 + It plays a role in the downregulation of antigen-dependent stimulation of T lymphocytes. Addition of anti-LAG-3 antibodies can result in T cell proliferation, high expression of activating antigens such as CD25, and increased concentrations of cytokines such as interferon-gamma and interleukin-4 (Huard et al. (1994) Eur. J. Immunol. 24:3216-3221). The cytoplasmic domain of LAG-3 can also interact with LAP, a signaling molecule involved in the downregulation of the CD3 / TCR activation pathway (Iouzalen et al. (2001) Eur. J. Immunol. 31:2885-2891). Furthermore, LAG-3 interacts with CD4 + CD25 + Regulatory T cells (T reg It contributes to the suppressor activity of ). reg Upon activation, cells express LAG-3, and antibodies against LAG-3 trigger T14. reg LAG-3 inhibits cellular suppression (Huang, C. et al. (2004) Immunity 21:503-513). LAG-3 can also negatively regulate T cell homeostasis in regulatory T cells through both T cell-dependent and T cell-independent mechanisms (Workman, CJ and Vignali, DA (2005) J. Immunol. 174:688-695). Therefore, LAG-3 inhibition can lead to an enhancement of the immune response.

[0427] Therefore, in some respects, the present invention provides a method for treating (e.g., mitigating or inhibiting) cancer or tumors in a subject. The method comprises administering to the subject a therapeutically effective dose of the anti-LAG-3 antibody molecule described herein, e.g., an anti-LAG-3 antibody molecule, alone or in combination with, for example, one or more agents or methods. In some embodiments, the anti-LAG-3 antibody molecule may be used alone for inhibiting the growth of cancerous tumors. Alternatively, the anti-LAG-3 antibody may be used in combination with one or more of the following, as described below: standard treatment (e.g., for cancer or infectious disorders), other antibodies, immunomodulators (e.g., activators of co-inhibitory molecules or inhibitors of inhibitory molecules); vaccines, e.g., therapeutic cancer vaccines; or other forms of cellular immunotherapy. In some embodiments, the anti-LAG-3 antibody molecule is administered in combination with, for example, a modulator of a co-inhibitory molecule (e.g., an agonist of a co-inhibitory molecule) or a modulator of an inhibitory molecule (e.g., an inhibitor of an immune checkpoint inhibitor), as described herein.

[0428] In one embodiment, the method is suitable for in vivo cancer treatment. To achieve antigen-specific enhancement of the immune response, an anti-LAG-3 antibody molecule can be administered together with the target antigen. When an antibody against LAG-3 is administered in combination with one or more drugs, the combination can be administered in any order or simultaneously.

[0429] Cancer type; Theranostic method In one embodiment, a method is provided for treating a subject, for example, a method for reducing or mitigating a hyperproliferative state or disorder in the subject (e.g., cancer), for example, a solid tumor, hematopoietic carcinoma, soft tissue tumor, or metastatic lesion. The method comprises administering to the subject one or more of the anti-LAG-3 antibody molecules described herein, alone or in combination with other therapeutic agents or treatments.

[0430] As used herein, the term “cancer” means all types of cancerous growth or carcinogenesis, metastatic tissue, or malignantly transformed cells, tissues, or organs, regardless of histopathological type or degree of invasiveness. Examples of cancerous disorders include, but are not limited to, solid tumors, hematopoietic cancers, soft tissue tumors, and metastatic lesions. Examples of solid tumors include malignant tumors of various organs, such as those affecting the liver, lungs, breasts, lymphatic system, digestive system (e.g., colon), genitourinary system (e.g., kidneys, urothelial cells), prostate, and pharynx, e.g., sarcomas and cell tumors (including adenocarcinoma and squamous cell carcinoma). Adenocarcinoma includes malignant tumors such as most colon cancers, rectal cancers, renal cell carcinomas, liver cancers, non-small cell lung cancers, small intestine cancers, and esophageal cancers. Squamous cell carcinoma includes malignant tumors such as those affecting the lungs, esophagus, skin, head and neck region, oral cavity, anus, and cervix. Metastatic lesions of the said cancers can also be treated or prevented using the methods and compositions of the present invention.

[0431] Representative cancers whose growth can be inhibited using the antibody molecules disclosed herein include cancers that are generally responsive to immunotherapy. Non-limiting examples of preferred cancers for treatment include melanoma (e.g., advanced stage (e.g., stage II-IV) melanoma or HLA-A2 positive melanoma), pancreatic cancer (e.g., advanced pancreatic cancer), solid tumors, breast cancer (e.g., metastatic breast cancer, breast cancer that does not express estrogen receptor, progesterone receptor or Her2 / neu 1, 2 or all, e.g., trinegative breast cancer), and renal cell carcinoma (e.g., advanced stage (e.g., stage IV) or metastatic renal cell carcinoma (MRCC)). Furthermore, refractory or recurrent malignancies can be treated using the antibody molecules described herein.

[0432] Other cancers that can be treated include, for example, solid tumors such as prostate cancer (e.g., hormone-refractory prostate cancer), colon cancer, lung cancer (e.g., non-small cell lung cancer), bone cancer, skin cancer, head and neck cancer (e.g., HPV-positive squamous cell carcinoma), cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, perianal cancer, gastric cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Merkel cell carcinoma, pediatric solid tumors, bladder cancer, kidney or ureteral cancer, renal pelvis cancer, central nervous system (CNS) neoplasms, tumor angiogenesis, spinal axial tumors, brainstem gliomas, pituitary adenomas, Kaposi's sarcoma, epidermal carcinoma or squamous cell carcinoma or hematopoietic malignancies such as Hodgkin lymphoma, non-Hodgkin lymphoma, and esophageal cancer. This includes cancers of the small intestine, endocrine cancers, thyroid cancers, parathyroid cancers, adrenal cancers, soft tissue sarcomas, urethral cancers, penile cancers, chronic or acute leukemias including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia (e.g., relapsed or refractory chronic lymphocytic leukemia), pediatric solid tumors, lymphocytic lymphomas, multiple myeloma, myelodysplastic syndromes, bladder cancers, kidney or ureteral cancers, renal pelvis cancers, central nervous system (CNS) neoplasms, primary CNS lymphomas, tumor angiogenesis, spinal axial tumors, brainstem gliomas, pituitary adenomas, Kaposi's sarcoma, epidermal carcinoma, squamous cell carcinoma, T-cell lymphomas, and environmentally induced cancers including those induced by asbestos (e.g., mesothelioma), as well as combinations thereof. Metastatic cancers, such as those expressing MHC class II molecules or LAG-3, can be treated using the antibody molecules described herein.

[0433] Without intending to be constrained by theory, in some embodiment, if a patient has cancer that highly expresses PD-L1 and / or if the cancer is infiltrated by antitumor immune cells, e.g., TILs, then the patient is likely to respond to treatment with anti-LAG-3, either alone or in combination with anti-PD-1 or PD-L1 antibody molecules (or, as desired, in combination with one or more agents as described herein). Antitumor immune cells are positive for CD8, PD-L1, and / or IFN-γ, and therefore the levels of CD8, PD-L1, and / or IFN-γ may be useful as indicators of the level of TILs in the tumor microenvironment. In some embodiment, this tumor microenvironment is triply positive for PD-L1 / CD8 / IFN-γ.

[0434] Therefore, in one respect, the present invention provides a method for determining whether a tumor sample is positive for one or more of PD-L1, CD8, and IFN-γ, and if the tumor sample is positive for one or more of these markers, for example, two or three in total, administering to the patient a therapeutically effective dose of an anti-PD-1 antibody molecule, optionally in combination with one or more other immunomodulatory agents or anticancer agents.

[0435] In the following indications, the majority of patients are triplicate-positive for PD-L1 / CD8 / IFN-γ: lung cancer (squamous cell); lung cancer (adenocarcinoma); head and neck cancer; gastric cancer; NSCLC; HNSCC; gastric cancer (e.g., MSIhi and / or EBV+); CRC (e.g., MSIhi); nasopharyngeal cancer (NPC); cervical cancer (e.g., squamous cell); thyroid cancer, e.g., papillary thyroid cancer; melanoma; TN breast cancer; and DLBCL (diffuse large B-cell lymphoma). Generally, in breast cancer and colon cancer, a significant proportion are triplicate-positive for PD-L1 / CD8 / IFN-γ. Also, in the indications for ER+ breast cancer and pancreatic cancer, a small proportion of patients are triplicate-positive for PD-L1 / CD8 / IFN-γ. These findings will be further examined in Example 4. Regardless of the proportion of patients who are positive for all three of these markers, screening patients for these markers allows for the determination of patient groups that are particularly likely to respond favorably to treatment with LAG-3 antibody alone or in combination with PD-1 antibody (e.g., blocking PD-1 antibody), optionally in combination with one or more other immunomodulatory agents (e.g., anti-TIM-3 antibody molecules or anti-PD-L1 antibody molecules) and / or anticancer agents, e.g., those listed in Table 7 and those disclosed in the publications listed in Table 7.

[0436] In one embodiment, cancer samples are classified as triplicate (positive for PD-L1 / CD8 / IFN-γ). This measurement is broadly divided into two thresholds: whether individual cells are classified as positive, and whether the sample as a whole is classified as positive. First, the levels of PD-L1, CD8, and / or IFN-γ can be measured within individual cells. In one embodiment, cells that are positive for one or more of these markers are cells with high levels of the markers compared to control cells or reference values. For example, in one embodiment, a high level of PD-L1 in a cell is higher than the level of PD-L1 in the corresponding non-cancerous tissue in the patient. As another example, in one embodiment, a high level of CD8 or IFN-γ in a cell is the level of this protein generally seen in TILs. Secondly, the percentage of cells in the sample that are positive for PD-L1, CD8, and / or IFN-γ can also be measured. (It is not necessary for a single cell to express all three markers.) In one embodiment, a triplicate sample is one that has a high percentage of cells that are positive for these markers, for example, those that are higher than the reference value or higher than the control sample.

[0437] In another embodiment, the overall levels of PD-L1, CD8, and / or IFN-γ can be measured in a sample. In this case, high levels of CD8 or IFN-γ in the sample may be levels of this protein generally found in cells infiltrated by TILs. Similarly, high levels of PD-L1 may be levels of this protein generally found in tumor samples, e.g., the tumor microenvironment.

[0438] Identifying a subset of patients who are triple-positive for PD-L1 / CD8 / IFN-γ, as shown in Example 4 below, reveals a subpopulation of patients who may be particularly responsive to PD-1 antibody therapy. For example, many IM-TN (immunomodulatory, triple-negative) breast cancer patients are triple-positive for PD-L1 / CD8 / IFN-γ. IM-TN breast cancer is described, for example, in Brian D. Lehmann et al., “Identification of human triple-negative breast cancer subtypes and preclinical models for selection of targeted therapies”, J Clin Invest. Jul 1, 2011; 121(7): 2750-2767. Triple-negative breast cancer does not express the estrogen receptor (ER), progesterone receptor (PR), and Her2 / neu. These cancers are difficult to treat because they generally do not respond to drugs targeting ER, PR, and Her2 / neu. Triple negative breast cancer can be further subdivided into different groups, one of which is immunomodulatory. As described by Lehmann et al., IM-TN breast cancer is rich in one or more genes involved in factors related to immune cell processes, such as immune cell signaling (e.g., TH1 / TH2 pathway, NK cell pathway, B cell receptor signaling pathway, DC pathway, and T cell receptor signaling), cytokine signaling (e.g., cytokine pathway, IL-12 pathway, and IL-7 pathway), antigen processing and presentation, signaling via core immune signaling pathways (e.g., NFKB, TNF, and JAK / STAT signaling), T cell function, immune transcription, interferon (IFN) response, and antigen processing.Therefore, in one embodiment, the cancer to be treated is a cancer that is positive or determined to be positive for one or more markers of IM-TN breast cancer, such as factors that promote immune cell signaling (e.g., TH1 / TH2 pathway, NK cell pathway, B cell receptor signaling pathway, DC pathway and T cell receptor signaling), cytokine signaling (e.g., cytokine pathway, IL-12 pathway and IL-7 pathway), antigen processing and presentation, signaling via core immune signaling pathways (e.g., NFKB, TNF and JAK / STAT signaling), T cell function, immune transcription, interferon (IFN) response and antigen processing.

[0439] As another example, a subset of colon cancer patients with high MSI (microsatellite instability) is also shown here to be triple-positive for PD-L1 / CD8 / IFN-γ. Therefore, in one embodiment, a LAG-3 antibody, e.g., the LAG-3 antibody described herein, alone or in combination with a PD-1 antibody (optionally in combination with a TIM-3 antibody or a PD-L1 antibody and one or more anticancer agents, e.g., one or more immunomodulators such as those listed in Table 7 or in publications in Table 7), is administered to patients with or identified as having high MSI colon cancer, thereby treating the cancer. In one embodiment, cells with high MSI are cells with higher levels of MSI than reference or control cells, e.g., non-cancerous cells of the same tissue type as the cancer.

[0440] As another example, it is shown here that a subset of gastric cancer patients who are high MSI and / or EBV-positive are also triplicate positive for PD-L1 / CD8 / IFN-γ. Therefore, in one embodiment, a LAG-3 antibody, e.g., the LAG-3 antibody described herein, alone or in combination with a PD-1 antibody (optionally in combination with a TIM-3 antibody or a PD-L1 antibody and one or more anticancer agents, e.g., one or more immunomodulators such as those listed in Table 7 or in the publications in Table 7), is administered to patients who have or have been identified as having high MSI and / or EBV-positive gastric cancer, thereby treating the cancer. In one embodiment, cells with high MSI are cells that have a higher level of MSI than reference or control cells, e.g., non-cancerous cells of the same tissue type as the cancer.

[0441] Disclosed herein are methods for assaying cancer for PD-L1 and then treating the cancer with a LAG-3 antibody, either alone or in combination with a PD-1 antibody. Cancer samples can be assayed at the PD-L1 protein level or mRNA level, as described in Example 5 herein. Samples having PD-L1 (protein or mRNA) levels higher than reference values ​​or control cells (e.g., non-cancerous cells) can be classified as PD-L1 positive. Thus, in one embodiment, a LAG-3 antibody, such as the LAG-3 antibody described herein, either alone or in combination with a PD-1 antibody (or optionally in combination with one or more anticancer agents) is administered to a patient who has or has been identified as having PD-L1 positive cancer. The cancer may be, for example, non-small cell lung (NSCLC) adenocarcinoma (ACA), NSCLC squamous cell carcinoma (SCC), or hepatocellular carcinoma (HCC).

[0442] In one embodiment, the method herein involves using a LAG-3 antibody, e.g., the LAG-3 antibody described herein, in combination with, for example, a PD-1 antibody, for the treatment of cancers that are positive for (or identified as positive for) PD-L1. In one embodiment, the cancers are colorectal cancer (e.g., high MSI), gastric cancer (e.g., high MSI and / or EBV+), NPC, cervical cancer, breast cancer (e.g., TN breast cancer), and ovarian cancer. In one embodiment, the cancers are NSCLC, melanoma, or HNSCC. In one embodiment, the LAG-3 antibody is administered in doses of, for example, 1, 3, 10, or 20 mg / kg.

[0443] For example, based on Example 4 below, it was found that certain gastric cancers that are triple-positive for PD-L1 / CD8 / IFN-γ are also positive for PIK3CA. Therefore, in one embodiment, cancer can be treated with LAG-3 antibodies, alone or in combination with anti-PD1 antibody molecules (optionally in combination with one or more immunomodulators, e.g., anti-TIM-3 antibody molecules or anti-PD-L1 antibody molecules) and PIK3CA inhibitors. Representative drugs in this category are described in Stein RC (September 2001). “Prospects for phosphoinositide 3-kinase inhibition as a cancer treatment”. Endocrine-related Cancer 8 (3): 237-48 and Marone R, Cmiljanovic V, Giese B, Wymann MP (January 2008). “Targeting phosphoinositide 3-kinase: moving towards therapy”. Biochimica et Biophysica Acta 1784 (1): 159-85.

[0444] For example, based on Example 4, patients with (or identified as having) high CRC, e.g., MSI high CRC, may be treated with LAG-3 antibody, alone or in combination with PD-1 antibody, optionally in combination with a treatment targeting one or both RNF43 and BRAF. For example, these cancers may be treated with LAG-3 antibody and PD-1 antibody, optionally in combination with one or more therapeutic agents targeting one or more of RNF43 and BRAF. In one embodiment, one or more therapeutic agents include anticancer agents listed in Table 7 or in the publications listed in Table 7. PD-1 inhibitors, e.g., antibodies, are described herein. RNF43 can be inhibited with, for example, antibodies, small molecules (e.g., 2-(2',3-dimethyl-[2,4'-bipyridine]-5-yl)-N-(5-(pyrazine-2-yl)pyridine-2-yl)acetamide (compound A28)), siRNA or Rspo ligand or derivatives thereof. BRAF inhibitors (e.g., vemurafenib or dabrafenib) are listed here.

[0445] For example, based on Example 4 below, a patient with (or identified as having) squamous cell lung cancer may be treated with a LAG-3 antibody molecule in combination with a PD-1-targeting therapeutic agent, such as a PD-1 antibody molecule, and optionally one or more anticancer agents, such as those listed in Table 7 or in publications in Table 7, or a TIM-3-targeting therapeutic agent, such as a TIM-3 antibody.

[0446] For example, based on Example 4 herein, patients with (or identified as having) thyroid cancer may be treated with a LAG-3 antibody molecule, alone or in combination with a PD-1 antibody molecule, optionally in combination with a BRAF-targeting agent, and optionally in combination with one or more immunomodulators, such as an anti-TIM-3 antibody molecule and an anti-PD-L1 antibody molecule. BRAF inhibitors (e.g., vemurafenib or dabrafenib) are described herein, for example, in Table 7 and the publications listed in Table 7.

[0447] In one aspect, the treatments herein may be used to treat patients who have (or have been identified as having) cancer associated with an infection, such as a viral or bacterial infection. Representative cancers include cervical cancer, anal cancer, HPV-associated head and neck squamous cell carcinoma, HPV-associated esophageal papilloma, HHV6-associated lymphoma, EBV-associated lymphoma (including Burkitt lymphoma), gastric MALT lymphoma, other infection-associated MALT lymphoma, HCC, and Kaposi's sarcoma. In another aspect, cancer includes, but is not limited to, hematopoietic malignancies or leukemia or lymphoma. For example, anti-LAG-3 antibody molecules are effective against acute leukemias including, but not limited to, B-cell acute lymphoblastic leukemia ("BALL"), T-cell acute lymphoblastic leukemia ("TALL"), and acute lymphoblastic leukemia (ALL); and one or more chronic leukemias including, but not limited to, chronic myeloid leukemia (CML) and chronic lymphocytic leukemia (CLL); for example, B-cell prelymphocytic leukemia, blastocyte plasmacytoid dendritic cell neoplasms, Burkitt lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, and small cell or large cell follicular lymphoma. It can be used to treat cancers and malignancies, including, but not limited to, further hematological malignancies or hematological conditions, including, but not limited to, “preleukemic conditions,” which is a diverse collection of hematopoietic conditions collectively defined by failure to produce (or dysplasia) malignant lymphoproliferative conditions, MALT lymphoma, mantle cell lymphoma, periphery zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndromes, non-Hodgkin lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasms, Waldenström macroglobulinemia, and myeloid hematological cells.

[0448] In one embodiment, the cancer is melanoma, e.g., advanced melanoma. In another embodiment, the cancer is advanced or unresectable melanoma that does not respond to other treatments. In yet another embodiment, the cancer is melanoma with a BRAF mutation (e.g., BRAF V600 mutation). In yet another embodiment, an anti-LAG-3 antibody molecule is administered after treatment with or without an anti-CTLA-4 antibody (e.g., ipilimumab) in combination with a BRAF inhibitor (e.g., vemurafenib or dabrafenib).

[0449] The methods and compositions described herein are useful for treating metastatic lesions associated with the aforementioned cancer.

[0450] Combination of anti-LAG-3 antibody and cancer vaccine LAG-3 can be combined with immunogenic agents such as cancer cells, purified tumor antigens (including recombinant proteins, peptides (e.g., HLA-A2 peptide) and carbohydrate molecules), cells, and cells transfected with genes encoding immunostimulatory cytokines (He et al. (2004) J. Immunol. 173:4919-28). Non-limiting examples of tumor vaccines that can be used include, for example, melanoma antigen peptides such as gp100 peptide, MAGE antigen, Trp-2, MART1 and / or tyrosinase or cytokine GM-CSF transfected tumor cells, DNA-based vaccines, RNA-based vaccines, and viral transduction-based vaccines. Cancer vaccines may be prophylactic or therapeutic.

[0451] LAG-3 blockade can be combined with vaccination protocols. Numerous experimental strategies have been devised for tumor vaccination (see Rosenberg, S., 2000, Development of Cancer Vaccines, ASCO Educational Book Spring: 60-62; Logothetis, C., 2000, ASCO Educational Book Spring: 300-302; Khayat, D. 2000, ASCO Educational Book Spring: 414-428; Foon, K. 2000, ASCO Educational Book Spring: 730-738; also see Restifo, N. and Sznol, M., Cancer Vaccines, Ch. 61, pp. 3023-3043 in DeVita, V. et al. (eds.), 1997, Cancer: Principles and Practice of Oncology. Fifth Edition). In one of these strategies, vaccines are prepared using autologous or allogeneic tumor cells. These cell vaccines have been shown to be most effective when tumor cells are transduced to express GM-CSF. GM-CSF has been shown to be a potent activator of antigen presentation for tumor vaccination (Dranoff et al. (1993) Proc. Natl. Acad. Sci. USA 90: 3539-43).

[0452] LAG-3 blockade can be combined with a collection of recombinant proteins and / or peptides expressed in tumors to produce an immune response to these proteins. These proteins are typically recognized by the immune system as autoantigens and are therefore tolerable to them. This may include protein telomerases, which are necessary for the synthesis of telomerase on tumor antigen chromosomes, expressed in over 85% of human cancers, and expressed in only a limited number of somatic tissues (Kim, N et al. (1994) Science 266: 2011-2013). (These somatic tissues can be protected from immune attack by various means). Tumor antigens can also be “neoantigens” expressed in cancer cells due to somatic mutations that alter protein sequences or create fusion proteins of two unrelated sequences (e.g., bcr-abl in the Philadelphia chromosome) or idiotypes from B-cell tumors.

[0453] Other tumor vaccines may contain proteins from viruses involved in human cancer, such as human papillomavirus (HPV), hepatitis viruses (HBV and HCV), Epstein-Barr virus (EBV), and Kaposi's herpes sarcoma virus (KHSV). Another form of tumor-specific antigen that can be used in combination with LAG-3 blockade is purified heat shock proteins (HSPs) isolated from tumor tissue itself. These heat shock proteins contain protein fragments from tumor cells, and these HSPs are highly effective for delivery to antigen-presenting cells to induce tumor immunity (Suot, R & Srivastava, P (1995) Science 269:1585-1588; Tamura, Y. et al. (1997) Science 278:117-120).

[0454] Dendritic cells (DCs) are potent antigen-presenting cells that can be used to induce antigen-specific responses. DCs can be produced ex vivo and can carry a variety of protein and peptide antigens as well as tumor cell extracts (Nestle, F. et al. (1998) Nature Medicine 4: 328-332). DCs can also be transduced by genetic means to express these tumor antigens as well. DCs have also been directly fused to tumor cells for immunization purposes (Kugler, A. et al. (2000) Nature Medicine 6:332-336). As a vaccination method, DC immunization can be effectively combined with LAG-3 blockade to activate a more p...

Claims

1. A pharmaceutical composition comprising an anti-LAG-3 antibody molecule for the treatment of cancer in a subject, The aforementioned pharmaceutical composition is formulated for use in combination with an alkylating agent, The aforementioned anti-LAG-3 antibody molecule is (a) A heavy chain variable region (VH) comprising the VHCDR1 amino acid sequence of SEQ ID NO: 4, the VHCDR2 amino acid sequence of SEQ ID NO: 5, and the VHCDR3 amino acid sequence of SEQ ID NO: 3; and a light chain variable region (VL) comprising the VLCDR1 amino acid sequence of SEQ ID NO: 13, the VLCDR2 amino acid sequence of SEQ ID NO: 14, and the VLCDR3 amino acid sequence of SEQ ID NO: 15; (b) VH, which includes the VHCDR1 amino acid sequence of SEQ ID NO: 1, the VHCDR2 amino acid sequence of SEQ ID NO: 2, and the VHCDR3 amino acid sequence of SEQ ID NO: 3; and VL, which includes the VLCDR1 amino acid sequence of SEQ ID NO: 10, the VLCDR2 amino acid sequence of SEQ ID NO: 11, and the VLCDR3 amino acid sequence of SEQ ID NO: 12; (c) VH including the VHCDR1 amino acid sequence of SEQ ID NO: 286, the VHCDR2 amino acid sequence of SEQ ID NO: 5, and the VHCDR3 amino acid sequence of SEQ ID NO: 3; and VL including the VLCDR1 amino acid sequence of SEQ ID NO: 13, the VLCDR2 amino acid sequence of SEQ ID NO: 14, and the VLCDR3 amino acid sequence of SEQ ID NO: 15; or (d) VH including the VHCDR1 amino acid sequence of SEQ ID NO: 286, the VHCDR2 amino acid sequence of SEQ ID NO: 2, and the VHCDR3 amino acid sequence of SEQ ID NO: 3; and VL including the VLCDR1 amino acid sequence of SEQ ID NO: 10, the VLCDR2 amino acid sequence of SEQ ID NO: 11, and the VLCDR3 amino acid sequence of SEQ ID NO: 12 A pharmaceutical composition containing the above.

2. The anti-LAG-3 antibody molecule, (a) VH containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and VL containing the amino acid sequence of SEQ ID NO: 32; (b) VH containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and VL containing the amino acid sequence of SEQ ID NO: 36; (c) VH containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and VL containing the amino acid sequence of SEQ ID NO: 40; (d) VH containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and VL containing the amino acid sequence of SEQ ID NO: 44; (e) VH containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and VL containing the amino acid sequence of SEQ ID NO: 48; (f) VH containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and VL containing the amino acid sequence of SEQ ID NO: 52; (g) VH containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and VL containing the amino acid sequence of SEQ ID NO: 56; (h) VH containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and VL containing the amino acid sequence of SEQ ID NO: 60; (i) VH containing the amino acid sequence of SEQ ID NO: 64 or SEQ ID NO: 104; and VL containing the amino acid sequence of SEQ ID NO: 36; (j) VH containing the amino acid sequence of SEQ ID NO: 64 or SEQ ID NO: 104; and VL containing the amino acid sequence of SEQ ID NO: 40; (k) VH containing the amino acid sequence of SEQ ID NO: 64 or SEQ ID NO: 104; and VL containing the amino acid sequence of SEQ ID NO: 56; (l) VH containing the amino acid sequence of SEQ ID NO: 64 or SEQ ID NO: 104; and VL containing the amino acid sequence of SEQ ID NO: 60; (m) VH containing the amino acid sequence of SEQ ID NO: 68 or SEQ ID NO: 108; and VL containing the amino acid sequence of SEQ ID NO: 36; (n) VH containing the amino acid sequence of SEQ ID NO: 72 or SEQ ID NO: 8; and VL containing the amino acid sequence of SEQ ID NO: 40; (o) VH containing the amino acid sequence of SEQ ID NO: 72 or SEQ ID NO: 8; and VL containing the amino acid sequence of SEQ ID NO: 60; (p) VH containing the amino acid sequence of SEQ ID NO: 76 and VL containing the amino acid sequence of SEQ ID NO: 60; (q) VH containing the amino acid sequence of SEQ ID NO: 80 and VL containing the amino acid sequence of SEQ ID NO: 84; (r) VH containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and VL containing the amino acid sequence of SEQ ID NO: 88; (s) VH containing the amino acid sequence of SEQ ID NO: 28 or SEQ ID NO: 100; and VL containing the amino acid sequence of SEQ ID NO: 92; or (t) VH containing the amino acid sequence of SEQ ID NO: 64 or SEQ ID NO: 104; and VL containing the amino acid sequence of SEQ ID NO: 96 A pharmaceutical composition according to claim 1, comprising:

3. The anti-LAG-3 antibody molecule, (a) A heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 34; (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 38; (c) A heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 42; (d) A heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 46; (e) A heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 50; (f) A heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 54; (g) A heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 58; (h) A heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 62; (i) a heavy chain containing the amino acid sequence of SEQ ID NO: 66 or SEQ ID NO: 106; and a light chain containing the amino acid sequence of SEQ ID NO: 38; (j) A heavy chain containing the amino acid sequence of SEQ ID NO: 66 or SEQ ID NO: 106; and a light chain containing the amino acid sequence of SEQ ID NO: 42; (k) A heavy chain containing the amino acid sequence of SEQ ID NO: 66 or SEQ ID NO: 106; and a light chain containing the amino acid sequence of SEQ ID NO: 58; (l) A heavy chain containing the amino acid sequence of SEQ ID NO: 66 or SEQ ID NO: 106; and a light chain containing the amino acid sequence of SEQ ID NO: 62; (m) A heavy chain containing the amino acid sequence of SEQ ID NO: 70 or SEQ ID NO: 110; and a light chain containing the amino acid sequence of SEQ ID NO: 38; (n) A heavy chain containing the amino acid sequence of SEQ ID NO: 74 or SEQ ID NO: 18; and a light chain containing the amino acid sequence of SEQ ID NO: 42; (o) A heavy chain containing the amino acid sequence of SEQ ID NO: 74 or SEQ ID NO: 18; and a light chain containing the amino acid sequence of SEQ ID NO: 62; (p) A heavy chain containing the amino acid sequence of SEQ ID NO: 78 and a light chain containing the amino acid sequence of SEQ ID NO: 62; (q) A heavy chain containing the amino acid sequence of SEQ ID NO: 82 and a light chain containing the amino acid sequence of SEQ ID NO: 86; (r) A heavy chain containing the amino acid sequence of SEQ ID NO: 30 or SEQ ID NO: 102; and a light chain containing the amino acid sequence of SEQ ID NO: 94; (s) A heavy chain containing the amino acid sequence of SEQ ID NO: 66 or SEQ ID NO: 106; and a light chain containing the amino acid sequence of SEQ ID NO: 98; (t) A heavy chain containing the amino acid sequence of SEQ ID NO: 113 and a light chain containing the amino acid sequence of SEQ ID NO: 34; (u) A heavy chain containing the amino acid sequence of SEQ ID NO: 113 and a light chain containing the amino acid sequence of SEQ ID NO: 38; (v) A heavy chain containing the amino acid sequence of SEQ ID NO: 122 and a light chain containing the amino acid sequence of SEQ ID NO: 38; (w) A heavy chain containing the amino acid sequence of SEQ ID NO: 122 and a light chain containing the amino acid sequence of SEQ ID NO: 58; or (x) Heavy chain containing the amino acid sequence of SEQ ID NO: 134 and light chain containing the amino acid sequence of SEQ ID NO: 38 A pharmaceutical composition according to claim 1, comprising:

4. The pharmaceutical composition according to claim 1, wherein the anti-LAG-3 antibody molecule comprises VH containing the amino acid sequence of SEQ ID NO: 104 and VL containing the amino acid sequence of SEQ ID NO:

56.

5. The pharmaceutical composition according to claim 1, wherein the anti-LAG-3 antibody molecule comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 122 and a light chain containing the amino acid sequence of SEQ ID NO:

58.

6. Pharmaceutical compositions, (a) A dose of 3 mg to 800 mg once a week, or once every two weeks, three weeks, or four weeks. (b) in doses of 3 mg, 20 mg, 80 mg, 240 mg or 800 mg, or (c) Once a week, once every two weeks, once every three weeks, or once every four weeks The pharmaceutical composition according to claim 1, formulated for administration.

7. A pharmaceutical composition according to any one of claims 1 to 6, wherein the alkylating agent is cisplatin.

8. The pharmaceutical composition according to any one of claims 1 to 6, wherein the alkylating agent is oxaliplatin.

9. The pharmaceutical composition according to any one of claims 1 to 8, wherein the cancer is a solid tumor, a hematopoietic carcinoma, a soft tissue tumor, or a metastatic lesion thereof.

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