EA040365B1 - TIM-3 ANTIBODY MOLECULES AND THEIR APPLICATIONS - Google Patents

Description

This application claims priority in US Provisional Application No. 61/934469, filed January 31, 2014, and US Provisional Application No. 62/094912, filed December 19, 2014, the contents of which are incorporated herein by reference in their entirety.

Sequence list

This application contains a listing of sequences, which was provided in electronic form in ASCII format and incorporated into the present description by reference in its entirety. The specified ASCII copy, created on January 26, 2015, is named C2160-7002WO_SL.txt and has a size of 208625 bytes.

State of the art

Activation of naive CD4+ T helper cells leads to the development of at least two distinct effector populations: Th1 cells and Th2 cells. See US 7470428, Mosmann T R et al. (1986) J Immunol 136:2348-57; Mosmann T R et al. (1996) Immunol Today 17:138-46; Abbas A K et al. (1996) Nature 383:787793. Th1 cells produce cytokines (eg, interferon-gamma, interleukin-2, tumor necrosis factor alpha, and lymphotoxin) that are commonly associated with cell-mediated immune responses against intracellular pathogens, delayed-type hypersensitivity reactions (Sher A et al. (1992) Annu Rev Immunol 10:385-409) and the induction of organ-specific autoimmune diseases. Liblau R S et al. (1995) Immunol Today 16:34-38. Th2 cells produce cytokines (eg, IL-4, IL-10 and IL-13) which are extremely important in controlling extracellular helminthic invasions and inducing atopic and allergic diseases. SherA et al. (1992) Annu Rev Immunol 10:38 5-409. In addition to their differing roles in disease, Th1 and Th2 cells cross-regulate each other's expansion and function. Thus, the preferential induction of Th2 cells inhibits autoimmune diseases (Kuchroo VK et al. (1995) Cell 80:707-18; Nicholson L B et al. (1995) Immunity 3:397-405), and the predominant induction of Th1 cells can regulate the induction of asthma, atopy and allergies. LackG et al. (1994) J Immunol 152:2546-54; Hofstra C L et al. (1998) J Immunol 161:5054-60.

TIM-3 is a transmembrane receptor protein that is expressed, for example, on Th1 (T helper 1) CD4+ cells and cytotoxic CD8+ T cells that secrete IFN-γ. TIM-3 is generally not expressed on naive T cells, but is upregulated on activated effector T cells. TIM-3 plays a role in the regulation of immunity and tolerance in vivo (see Hastings et al., Eur J Immunol. 2009 Sep; 39(9):2492-501). There is a need in the art for new molecules that regulate TIM-3 function and the function of TIM-3 expressing cells.

The essence of the invention

The present description describes antibody molecules that bind to TIM-3 (T-cell immunoglobulin and mucin domain 3) with high affinity and specificity. Also provided are nucleic acid molecules encoding antibody molecules, expression vectors, host cells, and methods for producing antibody molecules. Also contemplated are immunoconjugates, polyspecific or bispecific antibody molecules, and pharmaceutical compositions containing antibody molecules. The anti-TIM-3 antibody molecules described herein can be used (alone or in combination with other agents or therapeutic regimens) for the treatment, prevention and/or diagnosis of immune disorders, cancer, infectious disease, Crohn's disease, sepsis, SIRS ( systemic inflammatory response syndrome) and glomerulonephritis. Thus, the present description describes compositions and methods for detecting TIM-3, as well as methods for treating various diseases, including cancer and immune disorders, using anti-TIM-3 antibody molecules.

Thus, in certain aspects, the present invention provides an antibody molecule (e.g., an isolated or recombinant antibody molecule) having one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 , 13, 14, 15, 16 or all) of the following properties (a), (b), (c), (d), (e), (f), (g), ^(h) , (i), (j), ^(k) , ^(l) , ^(m) , ⁽ⁿ⁾ , (o), ⁽ p) or (q):

(a) binds TIM-3, such as human TIM-3, with high affinity, such as a dissociation constant (KD) of less than about 100 nM, typically about 10 nM, and more often about 1-0.1 nm, or stronger, e.g., less than about 0.2, 0.16, 0.15, 0.1, 0.075, 0.05, or 0.042 nM, (b) substantially binds to non-human primate TIM-3, e.g., TIM-3 cynomolgus monkey, with a dissociation constant (KD) of less than about 100 nM, typically about 10 nM, and more specifically about 3-0.3 nM or greater, such as 1-0.1 nM or greater, such as less than approximately 1 nM, 0.75 nM, or 0.68 nM, (c) inhibits TIM-3 binding to a TIM-3 ligand, such as phosphatidylserine (PtdSer), HMGB1, or CEACAM-1, (d) enhances IFN-gamma secretion and/ or TNF-alpha and/or proliferation of T cells, eg CD4+ or CD8+ T cells, eg CD4+ T cells, stimulated with anti-CD3/CD28 antibody in the presence of IL-12, or in autologous culture assays T cells-DCs stimulated with anti-CD3/CD28 antibody,

- 1 040365 (e) enhances the activity of cytotoxic NK cells (natural killers) against target cells (for example, K562 cells), for example, in in vitro assays, (f) enhances the ability of macrophages or antigen-presenting cells to stimulate a T-cell response, for example, increase the secretion of IL -12 antigen presenting cells, (g) binds specifically to an epitope on TIM-3, e.g., the same or similar epitope as an epitope recognized by an antibody molecule described herein, e.g., a mouse anti-TIM-3 antibody molecule or a humanized antibody molecule against TIM-3, as described in the present description, for example, an antibody molecule according to table. 1-4, (h) shows the same or similar binding affinity or specificity, or both affinity and specificity, as the antibody molecule according to table. 1-4, (i) exhibits the same or similar binding affinity or specificity, or both affinity and specificity, as an antibody molecule (eg, heavy chain variable region and light chain variable region) described in Table 1-4. 1-4, (j) exhibits the same or similar binding affinity or specificity, or both affinity and specificity, as an antibody molecule (e.g., heavy chain variable region and light chain variable region) containing the amino acid sequence shown in Table 1-4. 1-4, (g) binds specifically to an epitope on TIM-3, such as the same or a similar epitope as an epitope recognized by an antibody molecule described herein, such as a mouse anti-TIM-3 antibody molecule or a humanized anti-TIM-3 antibody molecule. TIM-3, as described in the present description, for example, an antibody molecule according to table. 1-4, (h) shows the same or similar binding affinity or specificity, or both affinity and specificity, as the molecule according to table. 1-4, (i) exhibits the same or similar binding affinity or specificity, or both affinity and specificity, as the molecule (eg, heavy chain variable region and light chain variable region) described in Table 1-4. 1-4, (j) exhibits the same or similar binding affinity or specificity, or both affinity and specificity, as an antibody molecule (e.g., heavy chain variable region and light chain variable region) containing the amino acid sequence shown in Table 1-4. 1-4, (k) inhibits, for example competitively inhibits, the binding of a second antibody molecule to TIM-3, where the second antibody molecule is an antibody molecule described in the present description, for example an antibody molecule selected from table. 1-4, (l) binds the same (or substantially the same) or overlapping (or substantially overlapping) epitope as a second anti-TIM-3 antibody molecule, wherein the second antibody molecule is the antibody molecule described herein , for example, an antibody molecule selected from the table. 1-4, (m) competes for binding and/or binds the same (or substantially the same) or overlapping (or substantially overlapping) epitope as a second anti-TIM-3 antibody molecule, wherein the second antibody molecule is a antibodies described in the present description, such as an antibody molecule selected from the table. 1-4, for example, as determined by the methods described in example 11, (n) has one or more biological properties of the antibody molecule described in the present description, such as an antibody molecule selected from table. 1-4, (o) has one or more pharmacokinetic properties of the antibody molecule described in the present description, such as an antibody molecule selected from table. 1-4, (p) modulates (eg, enhances or inhibits) one or more TIM-3 activities, for example results in one or more of: increased secretion of IFN-gamma and/or TNF-alpha in T cells; increased proliferation in T cells, such as CD4+ or CD8+ T cells; increased cytotoxic activity of NK cells; decrease in the suppressor activity of regulatory T cells (Treg); or enhancing the immunostimulatory properties of macrophages and/or antigen-presenting cells, such as enhancing the secretion of cytokines, such as the secretion of IL-12; or (q) binds to one or more of: two residues adjacent to the N-terminus of the A chain (residues Val24 and Glu25 in human TIM-3), loop BC, loop CC', F chain, loop FG and G TIM-3 chains, or one or more residues from a combination of two, three, four, five, or all of: two residues adjacent to the N-terminus of the A chain (Val24 and Glu25 residues in human TIM-3), BC loops, CC' loops, F chains, FG loops and TIM-3 G chains, for example, where binding is analyzed using ELISA or Biacore.

In some embodiments, the antibody molecule binds TIM-3 with high affinity, such as a KD that is at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% lower than the KD of a mouse anti-TIM-3 antibody molecule, such as the mouse anti-TIM-3 antibody molecule described herein.

In some embodiments, the level of expression of the TIM-3 antibody molecule is higher, such as at least about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times

- 2 040365 higher than the level of expression of a murine antibody molecule, such as a TIM-3 anti-mouse antibody molecule or a chimeric anti-TIM-3 antibody molecule described herein. In some embodiments, the antibody molecule is expressed in mammalian cells, such as rodent cells.

In some embodiments, the anti-TIM-3 antibody molecule reduces one or more TIM-3 activities with an IC ₅₀ (concentration at 50% inhibition) that is lower, e.g., at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% lower than the IC ₅₀ of a mouse anti-TIM-3 antibody molecule, such as the mouse anti-TIM-3 antibody molecule described herein. In some embodiments, TIM-3 activity is TIM-3 binding to one or more (e.g., one, two, three, four, or all) of the TIM-3 ligands described herein, e.g., one, two, or more (all ) from PtdSer, CEACAM-1 or HMGB1.

In some embodiments, an anti-TIM-3 antibody molecule interacts with, e.g., binds, surface (e.g., one, two, three, five, eight, ten, fifteen, or more) continuous or discontinuous (e.g., non-contiguous) amino acid residues of TIM-3, selected from Val24, Glu25, Thr41, Gly56, Ala57, Cys58, Pro59, Val60, Phe61, Glu121, Lys122, Phe123, Asn124, Leu125, Lys126 and/or Leu127.

In some embodiments, an anti-TIM-3 antibody molecule interacts with, e.g., binds, surface (e.g., one, two, three, five, eight, ten, fifteen, or more) continuous or discontinuous (e.g., non-contiguous) amino acid residues of TIM-3, selected from Val24, Glu25, Tyr26, Phe39, Tyr40, Thr41, Gly56, Ala57, Cys58, Pro59, Val60, Phe61, Ser105, Gly106, Ile107, Asn119, Asp120, Glu121, Lys122, Phe123, Asn124, Leu125, Lys126, Leu127 and /or Val128, for example, as detailed in the table. 13.

In some embodiments, an anti-TIM-3 antibody molecule interacts with, e.g., binds, surface (e.g., one, two, three, five, eight, ten, fifteen, or more) continuous or discontinuous (e.g., non-contiguous) amino acid residues of TIM-3, selected from Glu23, Val24, Glu25, Tyr26, Thr41, Pro42, Ala43, Ala44, Pro45, Gly46, Asn47, Leu48, Val49, Pro50, Val51, Cys52, Trp53, Gly54, Lys55, Gly56, Ala57, Cys58, Pro59, Val60, Phe61, Glu121, Lys122, Phe123, Asn124, Leu125, Lys126 and/or Leu127.

In some embodiments, an anti-TIM-3 antibody molecule interacts with, e.g., binds, surface (e.g., one, two, three, five, eight, ten, fifteen, or more) continuous or discontinuous (e.g., non-contiguous) amino acid residues of TIM-3, selected from Val24, Glu25, Tyr26, Phe39, Tyr40, Thr41, Pro42, Ala43, Ala44, Pro45, Gly46, Asn47, Leu48, Val49, Pro50, Val51, Cys52, Trp53, Gly54, Lys55, Gly56, Ala57, Cys58, Pro59, Val60, Phe61, Ser105, Gly106, Ile107, Asn119, Asp120, Glu121, Lys122, Phe123, Asn124, Leu125, Lys126, Leu127 and/or Val128.

In other embodiments, the anti-TIM-3 antibody molecule competes with CEACAM-1 for binding to TIM-3. In one embodiment, the anti-TIM-3 molecule interacts with, eg, binds, one, two, or all of Cys58, Asn119, and Lys122 of TIM-3, eg, displaces or competes with CEACAM-1 for binding to these residues. In one embodiment, the anti-TIM-3 antibody molecule reduces or blocks hydrogen bond formation between TIM-3 Lys122 and CEACAM-1 Asn42, e.g., at least about 10%, 20%, 30%, 40%, 50%, 60 %, 70%, 80% or 90% compared to hydrogen bonding between Lys122 TIM-3 and Asn42 CEACAM-1 in the absence of an anti-TIM-3 antibody molecule.

In another embodiment, the anti-TIM-3 antibody molecule interacts with, eg binds to, the PtdSer-binding loop of TIM-3. In one embodiment, the anti-TIM-3 antibody molecule interacts with, e.g., binds, at least two TIM-3 PtdSer-binding loops, e.g., the FG loop and the TIM-3 CC' loop (e.g., metal ion dependent ligand binding site (MILIBS) ). For example, the carboxyl group of PtdSer can bind to the CC' loop of TIM-3 and the amino group of PtdSer can bind to the FG loop of TIM-3. In one embodiment, the anti-TIM-3 antibody molecule reduces or prevents PtdSer-mediated membrane permeation of TIM-3.

In another embodiment, the anti-TIM-3 antibody molecule competes with HMGB1 for binding to TIM-3. For example, it reduces HMGB1 binding to TIM-3 residue 62 (Q in mouse TIM-3, E in human TIM-3), e.g., at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% compared to HMGB1 binding to TIM-3 residue 62 in the absence of an anti-TIM-3 antibody molecule.

In another embodiment, the anti-TIM-3 antibody molecule does not compete with the ligand galectin-9 (Gal-9) for binding to TIM-3.

In some embodiments, the anti-TIM-3 antibody molecule has increased stability, e.g., is at least about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times more stable in vivo, or in vitro than an anti-mouse TIM-3 antibody molecule or a humanized anti-TIM-3 antibody molecule, e.g. an anti-mouse TIM-3 antibody molecule or humanized

- 3 040365 antibody molecule against TIM-3 described in the present description.

In some embodiments, an anti-TIM-3 antibody molecule comprises at least one antigen-binding region, e.g., a variable region or an antigen-binding fragment thereof, from an antibody described herein, e.g., an antibody selected from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02 , ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3 -hum16, ABTIM3-hum17, ABTIM3hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Table. 1-4; or encoded by the nucleotide sequence shown in table. 1-4; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to the above sequences.

In certain embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, three, or four variable regions from an antibody described herein, e.g., an antibody selected from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3hum03, ABTIM3-hum04 ABTIM3-hum05 ABTIM3-hum06 ABTIM3-hum07 ABTIM3-hum08 ABTIM3hum09 ABTIM3-hum10 ABTIM3-hum11 ABTIM3-hum12 ABTIM3-hum13 ABTIM3-hum14 ABTIM3hum15 ABTIM3-hum16 ABTIM3- hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Table. 1-4; or encoded by the nucleotide sequence shown in table. 1-4; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to the above sequences.

In some embodiments, an anti-TIM-3 antibody molecule comprises at least one or two heavy chain variable regions from an antibody described herein, e.g., an antibody selected from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3hum03, ABTIM3- hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Table. 1-4; or encoded by the nucleotide sequence shown in table. 1-4; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to the above sequences.

In certain embodiments, an anti-TIM-3 antibody molecule comprises at least one or two light chain variable regions from an antibody described herein, e.g., an antibody selected from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3hum03, ABTIM3- hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Table. 1-4; or encoded by the nucleotide sequence shown in table. 1-4; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to the above sequences.

In one embodiment, the anti-TIM-3 antibody molecule comprises a heavy chain constant region for IgG4, eg, human IgG4. In another embodiment, human IgG4 comprises a substitution (e.g., substitution of Ser to Pro) at position 228 according to EU numbering or at position 108 of SEQ ID NO: 108 or 110. In another embodiment, the anti-TIM-3 antibody molecule comprises a heavy chains for IgG1, for example human IgG1. In one embodiment, human IgG1 comprises a substitution (e.g., substitution of Asn to Ala) at position 297 according to EU numbering or position 180 of SEQ ID NO: 112. In one embodiment, human IgG1 comprises a substitution (e.g., substitution of Asp to Ala) at position 265 according to EU numbering or at position 148 of SEQ ID NO: 113, a replacement (e.g. replacing Pro with Ala) at position 329 according to EU numbering, or at position 212 of SEQ ID NO: 113, or both of them. In one embodiment, human IgG1 comprises a substitution (e.g., substitution of Leu to Ala) at position 234 according to EU numbering or position 117 of SEQ ID NO: 114, a substitution (e.g., substitution of Leu to Ala) at position 235 according to EU numbering EU, or at position 118 of SEQ ID NO: 114, or both. In one embodiment, the heavy chain constant region contains the amino acid sequence shown in table. 1-5, or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to it.

In another embodiment, the anti-TIM-3 antibody molecule comprises a kappa light chain constant region, eg, a human kappa light chain constant region. In one embodiment, the light chain constant region contains the amino acid sequence shown in table. 1-5, or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to it.

- 4 040365

In another embodiment, the anti-TIM-3 antibody molecule comprises an IgG4 heavy chain constant region, such as human IgG4, and a kappa light chain constant region, such as a human kappa light chain constant region, such as a heavy and light chain constant region, comprising the amino acid sequence specified in tab. 1-5, or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to it. In another embodiment, the anti-TIM-3 antibody molecule comprises an IgG1 heavy chain constant region, such as human IgG1, and a kappa light chain constant region, such as a human kappa light chain constant region, such as a heavy and light chain constant region, comprising the amino acid sequence specified in tab. 1-5, or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to it. In one embodiment, human IgG1 includes a substitution at position 297 according to EU numbering (eg, Asn to Ala substitution). In one embodiment, human IgG1 comprises a substitution at position 265 according to EU numbering, a substitution at position 329 according to EU numbering, or both (e.g., substitution of Asp to Ala at position 265 and/or substitution of Pro to Ala at position 329 ). In one embodiment, the human IgG1 comprises a substitution at position 234 according to EU numbering, a substitution at position 235 according to EU numbering, or both (e.g., a change from Leu to Ala at position 234 and/or a change from Leu to Ala at position 235 ).

In another embodiment, the anti-TIM-3 antibody molecule comprises a heavy chain variable domain and constant domain, a light chain variable domain and constant region, or both, comprising the amino acid sequence of ABTIM3, ABTIM3-hum01, ABTIM3-hum02,

ABTIM3-hum03,

ABTIM3-hum09,

ABTIM3-hum15,

ABTIM3-hum04,

ABTIM3-hum10,

ABTIM3-hum16,

ABTIM3-hum05,

ABTIM3-hum11,

ABTIM3-hum17,

ABTIM3-hum06,

ABTIM3-hum12,

ABTIM3-hum18,

ABTIM3-hum07,

ABTIM3-hum13,

ABTIM3-hum19,

ABTIM3-hum08,

ABTIM3-hum14,

ABTIM3-hum20,

ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Table. 1-4; or encoded by the nucleotide sequence shown in table. 1-4; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the above sequences.

In some embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, or three complementarity determining regions (CDRs) from the heavy chain variable region of an antibody described herein, e.g., an antibody selected from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02 ABTIM3-hum03 ABTIM3-hum04 ABTIM3-hum05 ABTIM3hum06 ABTIM3-hum07 ABTIM3-hum08 ABTIM3-hum09 ABTIM3-hum10 ABTIM3-hum11 ABTIM3hum12 ABTIM3-hum13 ABTIM3-hum14 ABTIM3- hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Table. 1-4; or encoded by the nucleotide sequence shown in table. 1-4; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the above sequences.

In some embodiments, an anti-TIM-3 antibody molecule contains at least one, two, or three complementarity determining regions (CDRs) from a heavy chain variable region comprising the amino acid sequence shown in Table 1. 1-4, or encoded by the nucleotide sequence shown in table. 1-4. In one embodiment, one or more CDRs (or collectively all CDRs) have one, two, three, four, five, or more changes, such as amino acid substitutions, insertions, or deletions, relative to the amino acid sequence shown in Table 1. 1-4, or encoded by the nucleotide sequence shown in table. 1-4. In certain embodiments, the anti-TIM-3 antibody molecule comprises a heavy chain CDR substitution, eg one or more heavy chain CDR1, CDR2 and/or CDR3 substitutions. In one embodiment, the anti-TIM-3 antibody molecule comprises a substitution in the heavy chain CDR2 at position 55 of the heavy chain region, such as a substitution of asparagine for serie, or asparagine for glutamine, at position 55 of the heavy chain region according to Table 1. 1-4 (eg, any of SEQ ID NOs: 1 or 18 for an unmodified mouse sequence or a humanized sequence; or any of SEQ ID NOs: 26 or 32 for a modified sequence).

In some embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, or three complementarity determining regions (CDRs) from the light chain variable region of an antibody described herein, e.g., an antibody selected from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02 ABTIM3-hum03 ABTIM3-hum04 ABTIM3-hum05 ABTIM3hum06 ABTIM3-hum07 ABTIM3-hum08 ABTIM3-hum09 ABTIM3-hum10 ABTIM3-hum11 ABTIM3hum12 ABTIM3-hum13 ABTIM3-hum14 ABTIM3- hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Table. 1-4, or encoded by the nucleotide sequence shown in table. 1-4; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the aforementioned sequences.

- 5 040365

In certain embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, or three CDRs (or, collectively, all of the CDRs) from a light chain variable region comprising the amino acid sequence shown in Table 1. 1-4 or encoded by the nucleotide sequence shown in table. 1-4. In some embodiments, one or more CDRs (or, collectively, all CDRs) have one, two, three, four, five, six, or more changes, such as amino acid substitutions, insertions, or deletions, relative to the CDRs shown in Table 1. 1-4, or encoded by the nucleotide sequence shown in table. 1-4. In some embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, or three CDRs (or, collectively, all CDRs) from a light chain variable region containing the amino acid sequence shown in Table 1. 1-4 or encoded by the nucleotide sequence shown in table. 1-4. In some embodiments, one or more CDRs (or, collectively, all CDRs) have one, two, three, four, five, six, or more changes, such as amino acid substitutions, insertions, or deletions, relative to the CDRs shown in Table 1. 1-4, or encoded by the nucleotide sequence shown in table. 1-4.

In some embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, three, four, five, or six CDRs (or, collectively, all of the CDRs) from the heavy and light chain variable region comprising the amino acid sequence shown in Table . 1-4 or encoded by the nucleotide sequence shown in table. 1-4. In some embodiments, one or more CDRs (or, collectively, all CDRs) have one, two, three, four, five, six, or more changes, such as amino acid substitutions, insertions, or deletions, relative to the CDRs shown in Table 1. 1-4 or encoded by the nucleotide sequence shown in table. 1-4.

In certain embodiments, an anti-TIM-3 antibody molecule comprises all six CDRs from an antibody described herein, e.g., an antibody selected from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05 . , ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Table. 1-4; or encoded by the nucleotide sequence shown in table. 1-4, or highly similar CDRs, such as CDRs that are identical or that have at least one amino acid change but no more than two, three, or four changes (eg, substitutions, deletions, or insertions, eg, conservative substitutions). In certain embodiments, an anti-TIM-3 antibody molecule may include any of the CDRs described herein. In certain embodiments, the anti-TIM-3 antibody molecule comprises a heavy chain CDR substitution, eg one or more heavy chain CDR1, CDR2 and/or CDR3 substitutions. In one embodiment, the anti-TIM-3 antibody molecule comprises a substitution in the heavy chain CDR2 at position 55 of the heavy chain region, e.g., a substitution of asparagine for serine or asparagine for glutamine, at position 55 of the heavy chain region according to Table 1. 1-4 (eg, any of SEQ ID NOs: 1 or 18 for an unmodified mouse sequence or a humanized sequence; or any of SEQ ID NOs: 26 or 32 for a modified sequence).

In some embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, or three CDRs according to Kabat et al. (e.g., at least one, two, or three CDRs as defined by Kabat as listed in Tables 1-4) from the heavy chain variable region of an antibody described herein, e.g., an antibody selected from any of ABTIM3, ABTIM3hum01, ABTIM3- hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Table. 1-4; or encoded by the nucleotide sequence shown in table. 1-4; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the above sequences; or which have at least one amino acid change but no more than two, three, or four changes (eg, substitutions, deletions, or insertions, such as conservative substitutions) relative to one, two, or three CDRs according to Kabat et al. shown in Table . 1-4.

In certain embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, or three CDRs according to Kabat et al. (e.g., at least one, two, or three CDRs as defined by Kabat as indicated in Tables 1-4) from the light chain variable region of an antibody described herein, e.g., an antibody selected from any of ABTIM, ABTIM3hum01, ABTIM3- hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Table. 1-4; or encoded by the nucleotide sequence shown in table. 1-4; or sequentially

- 6 040365 substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the above sequences; or which have at least one amino acid change but no more than two, three, or four changes (eg, substitutions, deletions, or insertions, such as conservative substitutions) relative to one, two, or three CDRs according to Kabat et al. shown in Table . 1-4.

In certain embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, three, four, five, or six CDRs according to Kabat et al. (e.g., at least one, two, three, four, five, or six CDRs as defined by Kabat as indicated in Tables 1-4) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody selected from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3- hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in tables 1-4; or encoded by the nucleotide sequence shown in table. 1-4; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the above sequences; or which have at least one amino acid change but no more than two three or four changes (eg, substitutions, deletions, or insertions, such as conservative substitutions) relative to one, two, three, four, five, or six CDRs according to Kabat et al. shown in Table. 1-4.

In some embodiments, the anti-TIM-3 antibody molecule includes all six CDRs of Kabat et al. (e.g., all six CDRs as defined by Kabat as listed in Tables 1-4) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody selected from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3- hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Table. 1-4; or encoded by the nucleotide sequence presented in Table. 1-4; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the above sequences; or which have at least one amino acid change, but no more than two, three, or four changes (eg, substitutions, deletions, or insertions, eg, conservative substitutions) relative to all six CDRs according to Kabat et al., shown in table. 14. In one embodiment, an anti-TIM-3 antibody molecule may include any of the CDRs described herein.

In some embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, or three Chothia hypervariable loops (e.g., at least one, two, or three Chothia hypervariable loops as listed in Tables 1-4) of heavy chain variable region of an antibody described herein, e.g., an antibody selected from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09 ABTIM3-hum10 ABTIM3hum11 ABTIM3-hum12 ABTIM3-hum13 ABTIM3-hum14 ABTIM3-hum15 ABTIM3-hum16 ABTIM3hum17 ABTIM3-hum18 ABTIM3-hum19 ABTIM3-hum20 ABTIM3-hum21 ABTIM3- hum22, ABTIM3hum23; or as described in Table. 1-4; or encoded by the nucleotide sequence shown in table. 1-4; or at least amino acids from these hypervariable loops that contact TIM-3; or which have at least one amino acid change but no more than two, three, or four changes (eg, substitutions, deletions, or insertions, such as conservative substitutions) relative to one, two, or three hypervariable loops according to Chothia et al. shown in Table . 1-4.

In certain embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, or three Chothia hypervariable loops (e.g., at least one, two, or three Chothia hypervariable loops, as listed in Tables 1-4) of the variable light chain region of an antibody described herein, e.g., an antibody selected from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3 -hum09, ABTIM3-hum10, ABTIM3hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22 , ABTIM3hum23; or as described in Table. 1-4; or encoded by the nucleotide sequence shown in table. 1-4; or at least amino acids from these hypervariable loops that contact TIM-3; or which have at least one amino acid change but no more than two, three, or four changes (eg, substitutions, deletions, or insertions, such as conservative substitutions) relative to one, two, or three hypervariable loops according to Chothia et al., is shown

- 7 040365 in the table. 1-4.

In certain embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, three, four, five, or six hypervariable loops (e.g., at least one, two, three, four, five, or six hypervariable loops as defined by Chothia, as indicated in Tables 1-4) from the heavy and light chain variable regions of an antibody described herein, e.g. an antibody selected from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3- hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Table. 1-4; or encoded by the nucleotide sequence shown in table. 1-4; or at least amino acids from these hypervariable loops that contact TIM-3; or which have at least one amino acid change but no more than two, three or four changes (e.g. substitutions, deletions or insertions, e.g. conservative substitutions) relative to one, two, three, four, five or six hypervariable loops according to Chothia et al., shown in tables 1-4.

In some embodiments, an anti-TIM-3 antibody molecule includes all six hypervariable loops (e.g., all six hypervariable loops as defined by Chothia as listed in Tables 1-4) of an antibody described herein, e.g., an antibody selected from any of ABTIM3 , ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3 -hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or highly similar hypervariable loops, eg hypervariable loops that are identical or that have at least one amino acid change but no more than two, three or four changes (eg substitutions, deletions or insertions, eg conservative substitutions); or which have at least one amino acid change, but no more than two, three, or four changes (eg, substitutions, deletions, or insertions, eg, conservative substitutions) relative to all six hypervariable loops according to Chothia et al., shown in table. 1-4. In one embodiment, an anti-TIM-3 antibody molecule may include a hypervariable loop as described herein.

In a further embodiment, an anti-TIM-3 antibody molecule comprises at least one, two, or three hypervariable loops that have the same canonical structures as the corresponding hypervariable loop of an antibody described herein, e.g., an antibody selected from any of ABTIM3 , ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3 -hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22,

ABTIM3-hum23, for example, the same canonical structures as at least loop 1 and/or loop 2 of the heavy and/or light chain variable domains of the antibody described herein. See, for example, Chothia et al., (1992) J. Mol. Biol. 227:799-817; Tomlinson et al., (1992) J. Mol. Biol. 227:776-798 for descriptions of the canonical structures of hypervariable loops. These structures can be determined by examining the tables described in these links.

In certain embodiments, an anti-TIM-3 antibody molecule comprises a combination of CDRs or hypervariable loops as defined by Kabat et al. and Chothia et al.

In one embodiment, an anti-TIM-3 antibody molecule comprises at least one, two, or three CDRs or hypervariable loops from the heavy chain variable region of an antibody described herein, e.g., an antibody selected from any of ABTIM3, ABTIM3hum01, ABTIM3-hum02, ABTIM3-hum03 ABTIM3-hum04 ABTIM3-hum05 ABTIM3-hum06 ABTIM3hum07 ABTIM3-hum08 ABTIM3-hum09 ABTIM3-hum10 ABTIM3-hum11 ABTIM3-hum12 ABTIM3hum13 ABTIM3-hum14 ABTIM3-hum15 ABTIM3- hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23, as defined by Kabat and Chothia (e.g., at least one, two, or three CDRs or hypervariable loops, as defined Kabat and Chothia as listed in Tables 1-4); or encoded by the nucleotide sequence shown in table. 1-4; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the aforementioned sequences; or which have at least one amino acid change but no more than two, three or four changes (e.g. substitutions, deletions or insertions, e.g. conservative substitutions) relative to one, two or three CDRs or hypervariable loops according to Kabat and/or Chothia, shown in table. 1-4.

In another embodiment, the anti-TIM-3 antibody molecule comprises at least one, two, or three CDRs or hypervariable loops from the light chain variable region of an antibody described herein, e.g., an antibody selected from any of ABTIM3, ABTIM3-hum01,

- 8 040365

ABTIM3-hum02,

ABTIM3-hum08,

ABTIM3-hum14,

ABTIM3-hum03,

ABTIM3-hum09,

ABTIM3-hum15,

ABTIM3-hum04,

ABTIM3-hum10,

ABTIM3-hum16,

ABTIM3-hum05,

ABTIM3-hum11,

ABTIM3-hum17,

ABTIM3-hum06,

ABTIM3-hum12,

ABTIM3-hum18,

ABTIM3-hum07,

ABTIM3-hum13,

ABTIM3-hum19,

ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23, as defined by Kabat and Chothia (e.g., at least one, two, or three CDRs or hypervariable loops, as defined by Kabat and Chothia, as listed in Tables 1-4 ); or encoded by the nucleotide sequence shown in table. 1-4; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or more identical) to any of the above sequences; or which have at least one amino acid change but no more than two, three or four changes (e.g. substitutions, deletions or insertions, e.g. conservative substitutions) relative to one, two or three CDRs or hypervariable loops according to Kabat and/or Chothia, shown in table. 1-4.

An anti-TIM-3 antibody molecule may contain any combination of CDRs or hypervariable loops as defined by Kabat and Chothia.

In some embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, or three Chothia hypervariable loops from the heavy chain variable region of an antibody described herein, such as the antibody shown in Table 1. 1-4, or at least amino acids from these hypervariable loops that contact TIM-3.

In some embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, or three Chothia hypervariable loops from the light chain variable region of an antibody described herein, such as the antibody shown in Table 1. 1-4, or at least amino acids from these hypervariable loops that contact TIM-3.

In some embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, or three hypervariable loops according to Kabat from the heavy chain variable region of an antibody described herein, such as an antibody according to Table 1. 1-4, or at least amino acids from these hypervariable loops that contact TIM-3.

In some embodiments, the anti-TIM-3 antibody molecule comprises at least one, two, or three Kabat hypervariable loops from the light chain variable region of an antibody described herein, such as the antibody shown in Table 1. 1-4, or at least amino acids from these hypervariable loops that contact TIM-3.

In certain embodiments, an anti-TIM-3 antibody molecule comprises at least one, two, three, four, five, or six hypervariable loops from the heavy and light chain variable regions of an antibody described herein, such as an antibody shown in Table 1. 1-4, or at least amino acids from these variable loops that contact TIM-3.

In certain embodiments, an anti-TIM-3 antibody molecule includes all six hypervariable loops from the heavy and light chain variable regions of an antibody described herein, such as an antibody according to Table 1. 1-4, or at least amino acids from these hypervariable loops that contact TIM-3, or at least amino acids from these hypervariable loops that contact TIM-3, or highly related hypervariable loops, e.g., hypervariable loops that are identical or which have at least one amino acid change but no more than two, three or four changes (eg substitutions, eg conservative substitutions, deletions or insertions).

In some embodiments, an anti-TIM-3 antibody molecule includes at least one, two, or three hypervariable loops that have the same canonical structures as the corresponding hypervariable loop of an antibody described herein, such as an antibody according to Table 1. 1-4, for example, the same canonical structures as at least loop 1 and/or loop 2 of the heavy and/or light chain variable domains of an antibody described herein. See, for example, Chothia et al., (1992) J. Mol. Biol. 227:799-817; Tomlinson et al., (1992) J. Mol. Biol. 227:776798 for descriptions of the canonical structures of hypervariable loops. These structures can be determined by examining the tables described in these links. In one embodiment, such as an embodiment comprising a variable region, a CDR (eg, CDR according to Chothia or CDR according to Kabat) or other sequence mentioned in the present description, for example in table. 1-4, an antibody molecule is a monospecific antibody molecule, a bispecific antibody molecule, or is an antibody molecule that contains an antigen-binding fragment of an antibody, such as a half-antibody or an antigen-binding fragment of a half-antibody. In certain embodiments, an antibody molecule is a bispecific antibody molecule having a first binding specificity for TIM-3 and a second binding specificity for PD-1, LAG-3, CEACAM (e.g., CEACAM-1, CEACAM-3 and/or CEACAM -5), PD-L1 or PD-L2.

In certain embodiments, a light or heavy chain variable framework region (e.g., a region spanning at least FR1, FR2, FR3, or FR4) of an antibody molecule

- 9 040365 TIM-3 tive can be selected from: (a) a light or heavy chain variable framework region comprising at least 80%, 85%, 87% 90%, 92%, 93%, 95%, 97%, 98% or preferably 100% amino acid residues from a light or heavy chain variable framework region, eg, light or heavy chain variable framework region residues from a mature human antibody, human germline sequence, or human consensus sequence; (b) a light or heavy chain variable framework region comprising 20% to 80%, 40% to 60%, 60% to 90%, or 70% to 95% of the amino acid residues from the light or heavy chain variable framework, for example, light or heavy chain variable framework residues from a mature human antibody, human germline sequence, or human consensus sequence, (c) a non-human framework (eg, a rodent framework); or (d) a non-human framework region that has been modified, eg to remove antigenic or cytotoxic determinants, eg deimmunized or partially humanized. In some embodiments, the light or heavy chain variable framework comprises the light or heavy chain variable framework sequence at least 70, 75, 80, 85, 87, 88, 90, 92, 94, 95, 96, 97, 98 , 99% identical to the framework regions of the VL or VH segment of the human germline gene.

In certain embodiments, an anti-TIM-3 antibody molecule comprises a heavy chain variable domain having at least one, two, three, four, five, six, seven, ten, fifteen, twenty, or more changes, such as amino acid substitutions, insertions, or deletions. in the amino acid sequence, for example the amino acid sequence of the FR region of the entire variable region, for example as shown in FIG. 1A. In some embodiments, an anti-TIM-3 antibody molecule comprises a heavy chain variable domain having one or more (e.g., all) of: A at position 2, Y at position 3, S at position 7, R at position 13, V at position 37, R at position 42, V at position 72, A at position 79, or F at position 95, for example, the amino acid sequence of FR of the entire variable region, for example, as shown in FIG. 1A. In some embodiments, an anti-TIM-3 antibody molecule comprises a heavy chain variable domain having 2, 3, 4, 5, 6, 7, 8, or 9 positions selected from: A at position 2, Y at position 3, S at position 7, R at position 13, V at position 37, R at position 42, V at position 72, A at position 79 or F at position 95 of the amino acid sequence of the antibody, for example, according to table. 1-4.

In certain embodiments (and optionally in combination with the heavy chain substitutions described herein, e.g., in the previous paragraph), the anti-TM-3 antibody molecule contains a light chain variable domain having at least one, two, three, four, five, six, seven, ten, fifteen, twenty or more amino acid changes, such as amino acid substitutions, insertions or deletions, in the amino acid sequence shown in table. 1-4, such as the amino acid sequence of the FR region of the entire variable region, such as that shown in FIG. 1b. In certain embodiments, an anti-TIM-3 antibody comprises a light chain variable domain having M at position 89 of the amino acid sequence of the antibody according to Table 1. 1-4.

In some embodiments, the heavy or light chain variable domain, or both, of the anti-TIM-3 antibody molecule comprises an amino acid sequence that is substantially identical to the amino acid sequence described herein, e.g., at least 80%, 85%, 90 %, 92%, 95%, 97%, 98%, 99% or more identical to the variable region of an antibody described herein, e.g. an antibody selected from ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3 -hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17 , ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Table. 1-4; or encoded by the nucleotide sequence shown in table. 1-4; or which differs by at least 1 or 5 residues, but less than 40, 30, 20, or 10 residues, from the variable region of an antibody described herein.

In certain embodiments, the heavy or light chain variable region, or both, of an anti-TIM-3 antibody molecule comprises an amino acid sequence encoded by a nucleic acid sequence described herein or a nucleic acid that hybridizes to a nucleic acid sequence described herein. description (eg, the nucleic acid sequence as shown in Tables 1-4), or its complementary sequence, eg, under low stringency, moderate stringency, or high stringency conditions, or other hybridization conditions described herein.

In certain embodiments, an anti-TIM-3 antibody molecule contains at least one, two, three, or four antigen-binding regions, such as variable regions, having an amino acid sequence as shown in Table 1. 1-4, or a sequence substantially identical thereto (e.g., a sequence of at least about 85%, 90%, 95%,

- 10 040365

99% or more identical to it or differing by no more than 1, 2, 5, 10 or 15 amino acid residues from the sequences presented in table. 1-4). In certain embodiments, an anti-TIM-3 antibody molecule comprises a VH and/or VL domain encoded by a nucleic acid having a nucleotide sequence that encodes an antibody shown in Table 1. 1-4, or a sequence substantially identical to any of the nucleotide sequences (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical to it or differing by no more than 3, 6, 15 , 30 or 45 nucleotides from the sequences shown in Tables 1-4).

In certain embodiments, the anti-TIM-3 antibody molecule contains at least one, two, or three (eg, all) CDRs from the heavy chain variable region having the amino acid sequence as shown in Table 1. 1-4, or a sequence substantially homologous to it (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical to it and / or having one, two, three or more substitutions, insertions or deletions, such as conservative substitutions). In some embodiments, an anti-TIM3 antibody molecule contains at least one, two, or three (eg, all) CDRs from a light chain variable region having an amino acid sequence as shown in Table 1. 1-4, or a sequence substantially homologous to it (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical to it and / or having one, two, three or more substitutions, insertions or deletions, such as conservative substitutions). In certain embodiments, the anti-TIM-3 antibody molecule comprises at least one, two, three, four, five, or six (eg, all) of the heavy and light chain variable region CDRs having the amino acid sequence as shown in Table 1. 1-4, or a sequence substantially homologous to it (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical to it and / or having one, two, three or more substitutions, insertions or deletions, such as conservative substitutions).

In some embodiments, an anti-TIM-3 antibody molecule contains at least one, two, or three (e.g., all) CDRs and/or hypervariable loops from a heavy chain variable region having the amino acid sequence of an antibody described herein, e.g., an antibody selected from any of ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12 ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23 . 1-4, or a sequence substantially identical to it (for example, a sequence at least about 85%, 90%, 95%, 99% or more identical to it and / or having one, two, three or more substitutions, insertions or deletions, such as conservative substitutions). In certain embodiments, an anti-TIM-3 antibody molecule contains at least one, two, or three (e.g., all) CDRs and/or hypervariable loops from a light chain variable region having the amino acid sequence of an antibody described herein, e.g., an antibody selected from any of hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, hum13, ABTIM3 -hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18,

ABTIM3ABTIM3ABTIM3ABTIM3hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23, as summarized in Table 1-4, or a sequence substantially identical to it (for example, a sequence at least about 85%, 90%, 95%, 99% or more identical to it and / or having one, two, three or more substitutions, insertions or deletions, such as conservative substitutions). In some embodiments, the implementation of the TIM-3 antibody molecule contains all six CDRs and/or hypervariable loops described in the present description, for example, described in table. 1-4.

In some embodiments, the antibody molecule has a variable region that is identical in sequence or that differs by 1, 2, 3, or 4 amino acids from the variable region described herein (e.g., the FR region described herein).

In some embodiments, the anti-TIM-3 antibody molecule is a full length antibody or a fragment thereof (eg, Fab, F(ab') ₂ , Fv, or single chain Fv fragment (scFv)). In certain embodiments, the anti-TIM-3 antibody molecule is a monoclonal antibody or an antibody with a single specificity. The anti-TIM-3 antibody molecule can also be a humanized, chimeric antibody molecule, a camelid antibody molecule, a shark antibody molecule, or an in vitro-produced antibody molecule. In some embodiments, the anti-TIM-3 antibody molecule is a humanized antibody molecule. The heavy and light chains of an anti-TIM-3 antibody molecule may be full length (for example, an antibody may include at least one or at least two complete heavy chains and at least one or at least two complete light chains) or may include an antigen binding fragment (e.g. Fab, F(ab') ₂ , Fv, single-stranded Fv fragment, one

- 11 040365 nodomain antibody, diantibody (dAb), bivalent or bispecific antibody or fragment thereof, single domain variant thereof or antibody of camelids).

In certain embodiments, the anti-TIM-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 for TIM-3 and a second binding specificity, e.g., a second binding specificity for PD-1, LAG-3, CEACAM (e.g., CEACAM-1, -3 and/or -5 ), PD-L1 or PD-L2. In one embodiment, the bispecific antibody molecule binds to TIM-3 and PD-1. In another embodiment, the bispecific antibody molecule binds to TIM-3 and LAG-3. In another embodiment, the bispecific antibody molecule binds to TIM-3 and CEACAM (eg, CEACAM-1, -3 and/or -5). In another embodiment, the bispecific antibody molecule binds to TIM-3 and CEACAM-1. In another embodiment, the bispecific antibody molecule binds TIM-3 and CEACAM-3. In another embodiment, the bispecific antibody molecule binds TIM-3 and CEACAM-5. In another embodiment, the bispecific antibody molecule binds TIM-3 and PD-L1. In another embodiment, the bispecific antibody molecule binds TIM-3 and PD-L2. Any combination of the above molecules can be converted into a polyspecific antibody molecule, e.g., a trispecific antibody that includes a first binding specificity for TIM-3 and a second and third binding specificity for one or more of: PD-1, LAG-3, CEACAM (e.g., CEACAM-1, -3 and/or -5), PD-L1 or PD-L2.

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

In certain embodiments, the anti-TIM-3 antibody molecule has a heavy chain constant region (Fc) selected from, for example, IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE heavy chain constant regions; in particular selected from, for example, heavy chain constant regions of IgG1, IgG2, IgG3, and IgG4, more specifically, heavy chain constant region of IgG1 or IgG2 (eg human IgG1 or IgG2). In some embodiments, the heavy chain constant region is human IgG1. In some embodiments, the anti-TIM-3 antibody molecule has a light chain constant region selected, for example, from kappa or lambda light chain constant regions, in some embodiments kappa (eg, human kappa). In some embodiments, the constant region is altered, e.g., by mutation to modify properties of the anti-TIM-3 antibody molecule (e.g., to increase or decrease one or more of: Fc receptor binding, antibody glycosylation, amount of cysteine residues, effector cellular function, or complement function). For example, the constant region can be 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 change Fc- receptor (eg, mutant positions correspond to positions 132 (M to Y), 134 (S to T), 136 (T to E), 313 (H to K) and 314 (N to F) SEQ ID NO: 108 or 110; or positions 135 (M to Y), 137 (S to T), 139 (T to E), 316 (H to K) and 317 (N to F) SEQ ID NO: 111, 112, 113 or 114). In another embodiment, the heavy chain constant region of an IgG4, eg, human IgG4, is mutated at position 228 according to EU numbering (eg, S to P), eg, as shown in Table 1. 5. In certain embodiments, the anti-TIM-3 antibody molecule comprises human IgG4 mutated at position 228 according to EU numbering (eg, S to P), for example, as shown in Table 1. 5; and a kappa light chain constant region, for example, as shown in Table 1. 5. In the following embodiment, a heavy chain constant region of an IgG1, e.g., human IgG1, is mutated at one or more of position 297 (e.g., N to A), position 265 (e.g., D to A), position 329 (e.g., P on A), position 234 (eg L on A) or position 235 (eg L on A), all in accordance with the EU numbering, for example, as shown in table. 5. In certain embodiments, the anti-TIM-3 antibody molecule comprises human IgG1 mutated at one or more of the above positions, for example, as shown in Table 1. 5; and a kappa light chain constant region, for example, as shown in Table 1. 5. In some embodiments, the anti-TIM-3 antibody molecule is a humanized antibody molecule.

In some embodiments, anti-TIM-3 antibody molecules comprise combinations of human frameworks or humanized frameworks with CDRs (complementarity determining regions).

The invention also provides an antibody molecule that competes with a monoclonal antibody, such as the antibody molecule described herein, for binding to human TIM-3.

In certain embodiments, the implementation of the monoclonal antibody contains:

- 12 040365 (a) a heavy chain variable region (VH) containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 10 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14;

(b) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 4 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7 and the amino acid sequence of VLCDR3 SEQ ID NO: 8;

(c) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 2; and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14;

(d) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 24 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7 and the amino acid sequence of VLCDR3 SEQ ID NO: 8;

(e) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 31 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14; or (f) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 30 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7, and the amino acid sequence of VLCDR3 SEQ ID NO: 8.

The invention also relates to an antibody molecule that binds to the same (or substantially the same) or overlapping (or substantially overlapping) epitope as a monoclonal antibody, such as the antibody molecule described herein to human TIM-3.

In certain embodiments, the implementation of the monoclonal antibody contains:

(a) a heavy chain variable region (VH) containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 10 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14;

(b) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 4 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7 and the amino acid sequence of VLCDR3 SEQ ID NO: 8;

(c) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 25 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14;

(d) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 24 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7 and the amino acid sequence of VLCDR3 SEQ ID NO: 8;

(e) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 31 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14; or (f) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3;

- 13 040365 the amino acid sequence of VHCDR2 SEQ ID NO: 30 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7, and the amino acid sequence of VLCDR3 SEQ ID NO: 8.

The invention also relates to a nucleic acid molecule that contains one or both nucleotide sequences that encode heavy and light chain variable regions, CDRs, hypervariable loops, anti-TIM-3 antibody framework regions as described herein. In certain embodiments, the nucleotide sequence that encodes an anti-TIM-3 antibody molecule is codon-optimized. For example, the invention relates to first and second nucleic acids encoding the heavy and light chain variable regions, respectively, of an anti-TIM-3 antibody molecule selected from one or more, for example, any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03 , ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3 -hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23, as summarized in Table 1-4, or a sequence substantially identical to them. For example, the nucleic acid may contain a nucleotide sequence, as indicated in table. 1-4, or a sequence substantially identical thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical to it, or differing by no more than 3, 6, 15, 30, or 45 nucleotides from the sequences shown in Table 14).

In some embodiments, nucleic acids are described comprising nucleotide sequences that encode the heavy and light chain variable regions and CDRs of anti-TIM-3 antibody molecules as described herein. For example, the invention relates to first and second nucleic acids encoding the variable regions of the heavy and light chains, respectively, of the anti-TIM-3 antibody molecule according to Table. 1-4 or a sequence substantially identical thereto. For example, the nucleic acid may contain a nucleotide sequence encoding an anti-TIM-3 antibody molecule according to Table. 1-4, or a sequence substantially identical to that nucleotide sequence (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical to it or differing by no more than 3, 6, 15, 30 or 45 nucleotides from the above nucleotide sequence.

In certain embodiments, the implementation of the nucleic acid may contain a nucleotide sequence encoding at least one, two or three CDRs or hypervariable loops from the variable region of the heavy chain having the amino acid sequence as listed in table. 1-4, or a sequence substantially homologous thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical and/or having one, two, three or more substitutions, insertions, or deletions, such as conservative substitutions).

In certain embodiments, the implementation of the nucleic acid may contain a nucleotide sequence encoding at least one, two or three CDRs or hypervariable loops from the variable region of the light chain having the amino acid sequence as listed in table. 1-4, or a sequence substantially homologous to it (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical to it and / or having one, two, three or more substitutions, insertions or deletions, such as conservative substitutions).

In some embodiments, the nucleic acid may comprise a nucleotide sequence encoding at least one, two, three, four, five, or six CDRs or hypervariable loops from the heavy and light chain variable regions having the amino acid sequence as shown in Table 1. 1-4, or a sequence substantially homologous to it (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical to it and / or having one, two, three or more substitutions, insertions or deletions, such as conservative substitutions).

In some embodiments, the anti-TIM-3 antibody molecule is isolated or recombinant.

In some aspects, the present invention relates to host cells and vectors containing the nucleic acids described in the present description. Nucleic acids can be in the same vector or in separate vectors in the same host cell or in different host cells. The host cell may be a eukaryotic cell, such as a mammalian cell, a yeast cell, or a prokaryotic cell, such as E. coli. For example, the mammalian cell may be a cultured cell or cell line. Exemplary mammalian cells include lymphocytic cell lines (eg, NS0), Chinese hamster ovary (CHO) cells, COS cells, oocytes, and cells from a transgenic animal, such as mammary epithelial cells.

In some aspects, the present invention relates to a method of providing an anti

- 14 040365 of the body described in the present description. The method may include: providing a TIM-3 antigen (eg, an antigen containing at least a portion of a TIM-3 epitope, eg, a TIM-3 IgV domain); obtaining an antibody molecule that specifically binds to the TIM-3 antigen; and evaluating whether the antibody molecule specifically binds to the TIM-3 antigen, or evaluating the effectiveness of the antibody molecule in modulating, eg stimulating or inhibiting, TIM-3 activity. In addition, the method may include administering the antibody molecule to an individual, such as a human or non-human animal.

In some aspects, the invention relates to compositions, such as pharmaceutical compositions, which include a pharmaceutically acceptable carrier, excipient or stabilizer, and at least one of the anti-TIM-3 antibody molecules described in the present description. In one embodiment, a composition, such as a pharmaceutical composition, comprises a combination of an antibody molecule and one or more agents, such as a drug or other antibody molecule, as described herein. In some embodiments, the antibody molecule is conjugated to a label or drug. In some embodiments, the compositions, eg pharmaceutical compositions, comprise a combination of an antibody molecule and a second agent, eg a drug, or two or more of the aforementioned antibody molecules, as further described herein.

Anti-TIM-3 antibody molecules described herein can inhibit, reduce, or neutralize one or more TIM-3 activities, for example, resulting in blockade or reduction of immune control of T cells or NK cells, or resumption of the immune response by modulating antigen presenting cells. In one embodiment, the antibody molecule results in one or more of: increased secretion of IFN-gamma and/or TNF-alpha in T cells; enhancing the proliferation of T cells, such as CD4+ or CD8+ T cells; enhancing the cytotoxic activity of NK cells or reducing the suppressive activity of regulatory T cells (Treg) or macrophages; or increasing the ability of macrophages or dendritic cells to stimulate an immune response. Thus, such antibody molecules can be used to treat or prevent disorders in which it is desirable to enhance the immune response in an individual.

Application of Anti-TIM-3 Antibody Molecules

The antibody molecules described herein can modulate (eg, enhance, stimulate, increase, inhibit, decrease, or neutralize) one or more TIM-3 activities. In some embodiments, the antibody molecule results in one or more of: increased secretion of IFN-gamma and/or proliferation of T cells, or increased cytotoxic activity of NK cells. For example, in some embodiments, an anti-TIM-3 antibody molecule increases IFN-gamma secretion by at least 16%, 18%, 20%, 22%, 24%, 26%, 28%, or 30%, such as in the assay according to Example 4. In certain embodiments, an anti-TIM-3 antibody molecule increases the cytotoxic activity of NK cells by at least about 10%, 20%, 30%, 40%, 60%, 80%, or 100%, for example, in the assay according to Example 5. For example, an anti-TIM-3 antibody molecule can increase the cytotoxic activity of NK cells to at least about 60% or 70% of target cells killed when E/T=5, to at least about 75% or 85% target cells killed when E/T=12, or up to at least about 85% or 95% target cells killed when E/T=25, for example, in the assay of Example 5.

In some aspects, a method is provided for modulating (eg, stimulating or inhibiting) an immune response in an individual. The method comprises administering to an individual an anti-TIM-3 antibody molecule as described herein (e.g., a therapeutically effective amount of an anti-TIM-3 antibody molecule), alone or in combination with one or more agents or procedures, e.g., in combination with other immunomodulatory agents), so that there is a modulation of the immune response in the individual. In some embodiments, the implementation of the antibody molecule enhances, stimulates or enhances the immune response in the individual. In some embodiments, the antibody molecule inhibits, reduces, or neutralizes an immune response in an individual.

The subject may be a mammal, such as a monkey, a primate, preferably a higher primate, such as a human (eg, a patient having or at risk of having a disorder described herein). In some embodiments, the implementation of the individual needs to enhance the immune response and in some embodiments, the implementation of the individual needs to inhibit the immune response. In one embodiment, the individual has a disorder or is at risk for a disorder described herein, such as a cancer or an infectious disorder as described herein. In certain embodiments, the individual is immunodeficient or at risk for immunodeficiency. For example, the subject is or has been subjected to chemotherapy treatment and/or radiation therapy. Alternatively, or in combination, the subject is immunodeficient or at risk of immunodeficiency as a result of infection.

In one aspect, a method of treatment is provided (for example, one or more of reducing,

- 15 040365 inhibiting or slowing the progression) of a malignant tumor or neoplasm in an individual. The method includes administering to an individual an anti-TIM-3 antibody molecule as described herein, for example, a therapeutically effective amount of an anti-TIM3 antibody molecule, alone or in combination with one or more agents or procedures. In certain embodiments, an anti-TIM-3 antibody molecule is administered in combination with a costimulatory molecule modulator (eg, a costimulatory molecule agonist) or an inhibitory molecule modulator (eg, an immune response checkpoint inhibitor), for example, as described herein.

The present invention also relates to a method for reducing or inhibiting the growth of cancer or neoplasm cells (e.g., treatment of cancer) in an individual, comprising administering to the individual an anti-TIM-3 antibody molecule described herein, for example, a therapeutically effective amount of an anti-TIM-3 antibody molecule. , alone or in combination with a second agent, such as an immunomodulator (eg, a PD1, PD-L1, LAG-3, or CEACAM-1 inhibitor (eg, an antibody) or a combination).

In certain embodiments, the cancer being treated with an anti-TIM-3 antibody molecule, alone or in combination with one or more immunomodulators, includes, but is not limited to, solid tumor, hematologic cancer (e.g., leukemia, lymphoma, myeloma, e.g., multiple myeloma) and metastatic lesions. In one embodiment, the cancer is a solid tumor. Examples of solid tumors include malignant tumors, such as sarcomas and carcinomas, such as adenocarcinomas of various organ systems such as the lung, breast, ovary, lymphoid organs, gastrointestinal tract (eg, large intestine), anal canal, genital organs, and genitourinary tract ( eg, kidney, urothelium, bladder cells, prostate), pharynx, CNS (eg, brain cells, neuronal or glial cells), head and neck, skin (eg, melanoma) and pancreas, and adenocarcinomas, which include malignant tumors such as colon cancer, rectal cancer, renal cell carcinoma, liver cancer, non-small cell lung cancer, small intestine cancer and cancer of the esophagus. Malignant may be an early, intermediate, late stage, or metastatic cancer.

In one embodiment, the cancer is selected from lung cancer (e.g. lung adenocarcinoma or non-small cell lung cancer (NSCLC) (e.g. NSCLC with squamous and/or non-squamous histology or NSCLC adenocarcinoma)), melanoma (e.g. extended melanoma), kidney cancer (eg, renal cell carcinoma), liver cancer (eg, hepatic cell carcinoma), myeloma (eg, multiple myeloma), prostate cancer, breast cancer (eg, breast cancer that does not express one, two, or all of the estrogen receptor, progesterone receptor or Her2/neu, eg, triple-negative breast cancer), ovarian cancer, colorectal cancer, pancreatic cancer, head and neck cancer (eg, head and neck squamous cell carcinoma (HNSCC), anal cancer, cancer stomach and esophagus (eg, squamous cell carcinoma of the esophagus), mesothelioma, nasopharyngeal cancer, thyroid cancer, cervical cancer, lymphopro a liferative disease (eg, post-transplant lymphoproliferative disease) or a hematological malignancy (eg, diffuse large B-cell lymphoma, T-cell lymphoma, B-cell lymphoma, or non-Hodgkin's lymphoma) or leukemia (eg, myeloid leukemia or lymphoid leukemia).

In another embodiment, the cancer is selected from carcinoma (eg, expanded or metastatic carcinoma), melanoma, or lung carcinoma, eg, non-small cell lung carcinoma.

In one embodiment, the cancer is a lung cancer, such as lung adenocarcinoma, non-small cell lung cancer, or small cell lung cancer.

In one embodiment, the cancer is a melanoma, such as an extended melanoma. In one embodiment, the cancer is an advanced or unresectable melanoma that has not responded to other therapies. In other embodiments, the cancer is a melanoma with a BRAF mutation (eg, BRAF V600 mutation). In other embodiments, the anti-TIM-3 antibody molecule is administered following treatment with an anti-CTLA-4 antibody (eg, ipilimumab) with or without a BRAF inhibitor (eg, vemurafenib or dabrafenib).

In another embodiment, the cancer is a liver carcinoma, such as an advanced liver carcinoma with or without viral infection, such as chronic viral hepatitis.

In another embodiment, the cancer is prostate cancer, such as advanced prostate cancer.

In another embodiment, the cancer is a myeloma, such as multiple myeloma.

- 16 040365

In another embodiment, the cancer is a kidney cancer, such as renal cell carcinoma (RCC) (eg, metastatic RCC, clear cell renal cell carcinoma (CCRCC), or renal papillary cell carcinoma).

In one embodiment, the tumor microenvironment has an increased level of PD-L1 expression. Alternatively, or in combination, the tumor microenvironment has increased expression of IFNy and/or CD8.

In some embodiments, the individual has or is identified as having a tumor that has one or more of a high level or expression of PD-L1, or as having tumor-infiltrating lymphocytes (TIL)+ (e.g., as having an increased amount of TIL), or both. . In certain embodiments, the individual has or is identified as having a tumor that has a high level or expression of PD-L1 and that is TIL+. In some embodiments, the methods described herein further include identifying the individual as having a tumor that has one or more of high levels or expression of PD-L1, or as being TIL+, or both. In certain embodiments, the methods described herein further include identifying the individual as having a tumor that has a high level or expression of PD-L1 and as being TIL+. In some embodiments, tumors that are TIL+ are positive for CD8 and IFNy. In some embodiments, the individual has or is identified as having a high percentage of cells that are positive for one, two, or more of PD-L1, CD8, and/or IFNy. In certain embodiments, the individual has or is identified as having a high percentage of cells that are positive for all of PD-L1, CD8, and IFNy.

In some embodiments, the methods described herein further include identifying an individual based on the presence of a high percentage of cells that are positive for one, two, or more of PD-L1, CD8, and/or IFNy. In certain embodiments, the methods described herein further include identifying an individual based on the presence of a high percentage of cells that are positive for all of PD-L1, CD8, and IFNy. In some embodiments, the individual has or is identified as having one, two or more of PD-L1, CD8, and/or IFNy and one or more of lung cancer, eg, squamous cell lung cancer or lung adenocarcinoma; head and neck cancer; squamous cell carcinoma of the cervix; stomach cancer; esophageal cancer; thyroid cancer; melanoma and/or oropharyngeal cancer (NPC). In certain embodiments, the methods described herein further comprise identifying an individual based on the presence of one, two, or more of PD-L1, CD8, and/or IFNy and one or more of a lung cancer, eg, squamous cell lung cancer or lung adenocarcinoma; head and neck cancer; squamous cell carcinoma of the cervix; stomach cancer; thyroid cancer; melanoma and/or nasopharyngeal cancer.

In some embodiments, the individual has or is identified as having a tumor that has one, two or more of a high level or expression of PD-1, a high level or expression of TIM-3, and/or a high level of infiltration of regulatory T cells into the tumor, e.g., an increased number or percentage of Tregs present in the tumor. In certain embodiments, the individual has, or is identified as having, a tumor that has a high level or expression of PD-1 and TIM-3 and a high level, eg, number, of regulatory T cells in the tumor. In some embodiments, the methods described herein further comprise identifying an individual based on one, two, or more of a high percentage of cells that are positive for PD-1, a high percentage of cells that are positive for TIM-3, and/or a high level of infiltration of regulatory T cells into the tumor, eg an increased number or percentage of Tregs present in the tumor. In some embodiments, the methods described herein further comprise identifying an individual based on one, two, or more of a high percentage of cells that are positive for PD-1, a high percentage of cells that are positive for TIM-3, and/or a high level of regulatory T cell infiltration into the tumor, eg, an increased number or percentage of Tregs present in the tumor, and one or more of a lung cancer, eg, non-small cell lung cancer (NSCLC); hepatocellular carcinoma, eg hepatocellular carcinoma; or ovarian cancer, such as ovarian carcinoma.

The methods and compositions described herein are useful in the treatment of metastatic lesions associated with the aforementioned cancers.

In further aspects, the present invention relates to a method of treating an infectious disease in a subject, comprising administering to the subject a therapeutically effective amount of an anti-TIM-3 antibody described herein, or an antigen-binding portion thereof, alone or in combination with one or more agents or techniques (e.g., one or more immunomodulatory agents).

- 17 040365

In addition, the present invention provides methods for enhancing an immune response to an antigen in an individual, comprising administering to the individual: (i) an antigen; and (ii) antibodies against TIM-3, or an antigen-binding portion thereof, such that the immune response to the antigen is enhanced in the individual. The antigen may be, for example, a tumor antigen, a viral antigen, a bacterial antigen, or an antigen from a pathogen.

The anti-TIM-3 antibody molecule can be administered to an individual systemically (e.g., orally, parenterally, subcutaneously, intravenously, rectally, intramuscularly, intraperitoneally, intranasally, transdermally, or by inhalation or intrapalmary insertion), topically, or by application to mucous membranes, such as mucosal membranes of the nose, pharynx and bronchial tubes.

The anti-TIM-3 antibody molecule may be used alone in unconjugated form, or it may be coupled to a substance, such as a cytotoxic agent or moiety (eg, a therapeutic agent); a compound that emits radiation; molecules of plant, fungal or bacterial origin; or a biological protein (eg, a protein toxin) or a particle (eg, a recombinant viral particle, eg, via a viral envelope protein). For example, an anti-TIM-3 antibody can be coupled to a radioactive isotope such as an α-, β- or γ-emitter or a β- and γ-emitter.

Dosages and therapeutic regimens of the anti-TIM-3 antibody molecule can be determined by the skilled artisan. In certain embodiments, the anti-TIM-3 antibody molecule is administered by injection (e.g., subcutaneously or intravenously) at a dose of about 1 to 30 mg/kg, e.g., about 5 to 25 mg/kg, about 10 to 20 mg/kg, about 1 to 5 mg/kg, or about 3 mg/kg. The dosing schedule may vary, for example, from once a week to once every 2, 3 or 4 weeks. In one embodiment, the anti-TIM-3 antibody molecule is administered at a dose of about 10 to 20 mg/kg biweekly.

The antibody molecules described herein are preferred for use in the methods described herein, although other anti-TIM-3 antibodies may be used instead of or in combination with an anti-TIM-3 antibody molecule of the invention.

Combination Therapy

The methods and compositions described herein may be used in combination with other therapies. In some embodiments, the methods described herein comprise administering to an individual an anti-TIM-3 antibody molecule as described herein in combination with a cytotoxic agent in an amount effective to treat or prevent said disorder. The antibody molecule and the cytotoxic agent may be administered simultaneously or sequentially.

Any combination and sequence of anti-TIM-3 antibody molecules and other treatments can be used. The anti-TIM-3 antibody molecule and/or other therapeutic agent may be administered during periods of active disorder or during periods of remission or less active disease. The anti-TIM-3 antibody molecule and other therapeutic agent may be administered prior to treatment, concurrently with treatment, after treatment, or during remission of the disorder.

In certain embodiments, the methods and compositions described herein are administered in combination with one or more other antibody molecules, chemotherapy, another anti-cancer agent (e.g., targeted anti-cancer therapy, gene therapy, viral therapy, RNA therapy, bone marrow transplantation, nanotherapy, or oncolytic drugs), cytotoxic agents, immune therapies (eg, cytokines or cellular immune therapy), surgical procedures (eg, lumpectomy or mastectomy), or radiation procedures, or a combination of any of the above. Additional therapy may be in the form of adjuvant or neoadjuvant therapy. In some embodiments, the complementary therapy is an enzymatic inhibitor (eg, a small molecule enzymatic inhibitor) or a metastatic inhibitor. Exemplary cytotoxic agents that can be administered in combination include antimicrotubulin agents, topoisomerase inhibitors, antimetabolites, mitosis inhibitors, alkylating agents, anthracyclines, vinca alkaloids, intercalating agents, agents capable of interfering with the signal transduction cascade, agents that stimulate apoptosis, proteasome inhibitors, and radiation therapy (eg, local irradiation or whole body irradiation (eg, gamma irradiation)). In other embodiments, the implementation of additional therapy is surgery, or radiation therapy, or a combination of both. In other embodiments, the adjunctive therapy is a therapy that targets one or more of the PI3K/AKT/mTOR cascades, an HSP90 inhibitor, or a tubulin inhibitor.

Alternatively, or in combination with the above combinations, the methods and administration of the compositions described herein may be carried out in combination with one or more of: immunomodulators (eg, an activator of a costimulatory molecule or an inhibitor of an inhibitory molecule, eg, an immune response checkpoint molecule); vaccines, for example

- 18 040365 measures, a therapeutic vaccine against a malignant tumor; or other forms of cellular immunotherapy.

Illustrative non-limiting combinations and uses of anti-TIM-3 antibody molecules include the following.

In certain embodiments, an anti-TIM-3 antibody molecule is administered in combination with a modulator of a co-stimulatory molecule or an inhibitory molecule, such as a co-inhibitory ligand or receptor.

In one embodiment, an anti-TIM-3 antibody molecule is administered in combination with a modulator, such as an agonist, of a costimulatory molecule. In one embodiment, the co-stimulatory molecule agonist is selected from an agonist (e.g., an antibody agonist or an antigen-binding fragment thereof, or a soluble fusion construct) 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 ligand.

In one embodiment, an anti-TIM-3 antibody molecule is administered in combination with an inhibitor of an inhibitory molecule (or immune response checkpoint inhibitor) selected from PD-1, PD-L1, PD-L2, CTLA-4, LAG-3, CEACAM ( eg CEACAM-1, -3 and/or -5), VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGFR-beta. Inhibition of an inhibitory molecule can be accomplished by inhibition at the DNA, RNA or protein level. In embodiments, an inhibitory nucleic acid (eg, dsRNA, siRNA, or shRNA) can be used to inhibit the expression of an inhibitory molecule. In other embodiments, the inhibitor of the inhibitory signal is a polypeptide, such as a soluble ligand, or an antibody or antigen-binding fragment thereof, that binds to an inhibitory molecule. In one embodiment, the inhibitor is a soluble ligand (eg, CTLA-4-Ig), or an antibody or antibody fragment that binds to PD-L1, PD-L2, or CTLA-4. For example, an anti-TIM-3 antibody molecule can be administered in combination with an anti-CTLA-4 antibody, e.g., ipilimumab, e.g., for the treatment of cancer (e.g., a cancer selected from: melanoma, e.g. metastatic melanoma; lung cancer, e.g. non-small cell lung carcinoma or prostate cancer). In one embodiment, the anti-TIM-3 antibody molecule is administered following treatment with an anti-CTLA-4 antibody (eg, ipilimumab) with or without a BRAF inhibitor (eg, vemurafenib or dabrafenib).

In another embodiment, an anti-TIM-3 antibody molecule is administered in combination with an anti-TIM-3 antibody or antigen-binding fragment thereof.

In another embodiment, the anti-TIM-3 antibody molecule is administered in combination with an anti-PD-1 antibody or antigen-binding fragment thereof.

In other embodiments, an anti-TIM-3 antibody molecule is administered in combination with an anti-TIM-3 antibody and an anti-TIM-3 antibody (or antigen-binding fragments thereof).

In another embodiment, an anti-TIM-3 antibody molecule is administered in combination with a CEACAM inhibitor (eg, a CEACAM-1, -3 and/or -5 inhibitor), eg, an anti-CEACAM antibody molecule. In another embodiment, an anti-TIM-3 antibody molecule is administered in combination with a CEACAM-1 inhibitor, such as an anti-CEACAM-1 antibody molecule. In another embodiment, an anti-TIM-3 antibody molecule is administered in combination with a CEACAM-3 inhibitor, such as an anti-CEACAM-3 antibody molecule. In another embodiment, an anti-TIM-3 antibody molecule is administered in combination with a CEACAM-5 inhibitor, such as an anti-CEACAM-5 antibody molecule.

The combination of antibodies described herein can be administered alone, eg as single antibodies or antigen-binding fragments thereof, or in a bound state, eg as a bispecific or trispecific antibody molecule. In one embodiment, a bispecific antibody is administered that comprises an anti-TIM-3 antibody molecule and an anti-PD-1, anti-CEACAM (e.g., anti-CEACAM-1, -3 and/or -5) or anti-TIM-3 antibody, or an antigen-binding antibody thereof. fragment. In certain embodiments, a combination of antibodies described herein is used to treat a cancer, eg, a cancer as described herein (eg, a solid tumor or a hematologic cancer).

In other embodiments, the anti-TIM-3 antibody molecule is administered in combination with a cytokine. The cytokine can be administered as a fusion molecule with an anti-TIM-3 antibody molecule or as separate formulations. In one embodiment, the TIM-3 antibody is administered in combination with one, two, three or more cytokines, eg, as a fusion molecule or as separate compositions. In one embodiment, the cytokine is an interleukin (IL) selected from one, two, three or more of IL-1, IL-2, IL-12, IL-15, or IL-21. In one embodiment, the bispecific antibody molecule has a first binding specificity for a first target (e.g., TIM-3), a second binding specificity for a second target (e.g., LAG-3 or PD-1), and optionally associated with an interleukin domain (e.g., , IL

- 19 040365

12), for example, full-length IL-12 or part of it. In certain embodiments, a combination of an anti-TIM-3 antibody molecule and a cytokine described herein is used to treat a cancer, eg, a cancer as described herein (eg, a solid tumor).

In certain embodiments, the anti-TIM-3 antibody molecule is administered in combination with an antibody specific for HLA C, eg, an antibody specific for immunoglobulin-like killer cell receptors (also referred to as an anti-KIR antibody). In certain embodiments, a combination of an anti-TIM-3 antibody molecule and an anti-KIR antibody is used to treat a cancer, eg a cancer as described herein (eg, a solid tumor, eg, an expanded solid tumor).

In one embodiment, the anti-TIM-3 antibody molecule is administered in combination with cellular immunotherapy (eg, Provenge® (eg, Sipuleucel-T)) and optionally in combination with cyclophosphamide. In certain embodiments, a combination of an anti-TIM-3 antibody molecule, Provenge®, and/or cyclophosphamide is used to treat a cancer, eg, a cancer as described herein (eg, prostate cancer, eg, advanced prostate cancer).

In another embodiment, the anti-TIM-3 antibody molecule is administered in combination with a vaccine, eg, a cancer vaccine (eg, dendritic cell-based renal carcinoma (DC-RCC) vaccine). In one embodiment, the vaccine is peptide-based, DNA-based, RNA-based, or antigen-based, or a combination thereof. In embodiments, the vaccine contains one or more peptides, nucleic acids (eg, DNA or RNA), antigens, or a combination thereof. In certain embodiments, the combination of an anti-TIM-3 antibody molecule and a DC-RCC vaccine is used to treat a cancer, e.g., a cancer as described herein (e.g., renal carcinoma, e.g., metastatic renal cell carcinoma (RCC) or clear cell renal cell carcinoma (CCRCC). )).

In another embodiment, the anti-TIM-3 antibody molecule is administered in combination with an adjuvant.

In another embodiment, the TIM-3 antibody molecule is administered in combination with chemotherapy and/or immunotherapy. For example, an anti-TIM-3 antibody molecule can be used to treat myeloma, alone or in combination with one or more of: chemotherapeutic or other anti-cancer agents (e.g., thalidomide analogs, e.g. lenalidomide), anti-PD-1 antibodies treated with a tumor antigen dendritic cells, fusion (eg, by electrofusion) of tumor cells and dendritic cells, or vaccination with an immunoglobulin idiotype produced by malignant plasma cells. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with an anti-TIM-3 antibody for the treatment of myeloma, eg, multiple myeloma.

In one embodiment, an anti-TIM-3 antibody molecule is used in combination with chemotherapy to treat lung cancer, eg, non-small cell lung cancer. In one embodiment, an anti-TIM-3 antibody molecule is used with standard lung cancer chemotherapy, eg NSCLC, eg platinum doublet therapy, to treat lung cancer. In other embodiments, an anti-TIM-3 antibody molecule is used in combination with an indolamine-pyrrole-2,3-dioxygenase (IDO) inhibitor (e.g., INCB24360) in an individual with advanced or metastatic cancer (e.g., a patient with metastatic and recurrent cancer). NSCLC).

In other embodiments, an anti-TIM-3 antibody molecule is used in combination with one or more of: an immune-based strategy (eg, interleukin-2 or interferon-alpha), a targeting agent (eg, a VEGF inhibitor such as an anti-VEGF monoclonal antibody); a VEGF tyrosine kinase inhibitor such as sunitinib, sorafenib, axitinib and pazopanib; an RNA-i inhibitor; or an inhibitor of a downstream mediator of VEGF signaling, eg the mammalian target of rapamycin (mTOR) inhibitor, eg everolimus and temsirolimus. Any of these combinations can be used to treat kidney cancer, eg, renal cell carcinoma (RCC) (eg, clear cell renal cell carcinoma (CCRCC)) or metastatic RCC.

In some embodiments, an anti-TIM-3 antibody molecule, eg, an anti-TIM-3 antibody molecule described herein, is used in combination with a MEK inhibitor (eg, a MEK inhibitor as described herein). In some embodiments, a combination of an anti-TIM-3 antibody and a MEK inhibitor is used to treat a cancer (eg, the cancer described herein). In some embodiments, the cancer being treated with the combination is selected from melanoma, colorectal cancer, non-small cell lung cancer, ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, hematologic malignancy, or

- 20 040365 renal cell carcinoma. In certain embodiments, the cancer comprises a BRAF mutation (eg, BRAF V600E mutation), wild-type BRAF, wild-type KRAS, or an activating KRAS mutation. The cancer may be an early, intermediate or late stage cancer.

In another embodiment, an anti-TIM-3 antibody molecule is used in combination with one, two, or all of oxaliplatin, leucovorin, or 5-FU (eg, FOLFOX co-treatment). Alternatively, or in combination, the combination further comprises a VEGF inhibitor (eg, a VEGF inhibitor as described herein). In some embodiments, a combination of an anti-TIM-3 antibody, FOLFOX co-treatment, and a VEGF inhibitor is used to treat a cancer (eg, the cancer described herein). In some embodiments, the cancer being treated with the combination is selected from melanoma, colorectal cancer, non-small cell lung cancer, ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, hematologic malignancy, or renal cell cancer. The cancer may be an early, intermediate or late stage cancer.

In other embodiments, the anti-TIM-3 antibody molecule is administered with a tyrosine kinase inhibitor (eg, axitinib) for the treatment of renal cell carcinoma and other solid tumors.

In other embodiments, the anti-TIM-3 antibody molecule is administered with an agent that targets the 4-1BB receptor (eg, an antibody that stimulates signaling through 4-1BB (CD-137), eg, PF-2566). In one embodiment, an anti-TIM-3 antibody molecule is administered in combination with a tyrosine kinase inhibitor (eg, axitinib) and an agent that targets the 41BB receptor.

An anti-TIM-3 antibody molecule may be associated with a substance, such as a cytotoxic agent or a moiety (eg, a therapeutic drug; a compound that emits radiation; molecules of plant, fungal, or bacterial origin; or a biological protein (eg, a protein toxin) or particle (for example, a recombinant viral particle, for example, through a viral envelope protein)). For example, an antibody may be associated with a radioactive isotope such as an α-, β- or γ-emitter or a β- and γ-emitter.

Additional combination therapies

The methods and compositions described herein (for example, anti-TIM-3 antibodies and methods using them) can be used in combination with other agents or therapeutic regimens, for example, a second drug selected from one or more of the agents listed in Table. 6. In one embodiment, the methods described herein comprise administering to an individual an anti-TIM-3 antibody molecule as described herein (optionally in combination with one or more inhibitors of PD-1, PD-L1, PD-L2, LAG -3, CEACAM (eg, CEACAM-1 and/or CEACAM-5) or CTLA-4) also include the introduction of a second drug selected from one or more of the funds listed in table. 6 in an amount effective to treat or prevent a disorder, such as a disorder as described herein, such as cancer. When administered in combination, an anti-TIM-3 antibody molecule, an additional agent (e.g., a second or third agent), or all of them, may be administered in an amount or dose that is greater than, lower than, or the same as the amount or dosage of each agent. alone, for example, as monotherapy. In certain embodiments, the administered amount or dosage of the anti-TIM-3 antibody, additional agent (e.g., second or third agent), or all of them is lower (e.g., at least 20%, at least 30%, at least at least 40% or at least 50%) than the amount or dosage of each agent used alone, for example, as monotherapy. In other embodiments, the amount or dosage of the anti-TIM-3 antibody, additional agent (e.g., second or third agent), or all of them that results in the desired effect (e.g., treatment of cancer) is lower (e.g., at least 20%, at least 30%, at least 40%, or at least 50% lower).

In other embodiments, the implementation of the second drug is selected from one or more of the funds listed in table. 6. In one embodiment, the cancer is selected from lung cancer (e.g., non-small cell lung cancer (NSCLC) (e.g., NSCLC with squamous and/or non-squamous histology or NSCLC adenocarcinoma)) or described in the publication shown in Table. 6. In some embodiments, the second drug is selected from one or more of: 1) a protein kinase C (PKC) inhibitor; 2) inhibitor of heat shock protein 90 (HSP90); 3) a phosphoinositide 3-kinase inhibitor (PI3K) and/or a rapamycin target (mTOR); 4) a cytochrome P450 inhibitor (eg, a CYP17 inhibitor or a 17-alpha-hydroxylase/C17-20 lyase inhibitor); 5) an iron chelating agent; 6) an aromatase inhibitor; 7) a p53 inhibitor, eg an inhibitor of the p53/Mdm2 interaction; 8) apoptosis inducer; 9) an angiogenesis inhibitor; 10) an aldosterone synthase inhibitor; 11) a smoothened receptor inhibitor (SMO); 12) prolactin receptor inhibitor

- 21 040365 (PRLR); 13) Wnt signal transduction inhibitor; 14) a CDK4/6 inhibitor; 15) fibroblast growth factor receptor 2 (FGFR2)/fibroblast growth factor receptor 4 (FGFR4) inhibitor; 16) an inhibitor of macrophage colony stimulating factor (M-CSF); 17) an inhibitor of one or more of c-KIT, histamine release, Flt3 (eg FLK2/STK1) or PKC; 18) an inhibitor of one or more of VEGFR-2 (eg, FLK-1/KDR), PDGFR-beta, c-KIT, or Raf kinase C; 19) a somatostatin agonist and/or a growth hormone releasing inhibitor; 20) an anaplastic lymphoma kinase (ALK) inhibitor; 21) inhibitor of insulin-like growth factor receptor 1 (IGF-1R); 22) P-glycoprotein 1 inhibitor; 23) vascular endothelial growth factor receptor (VEGFR) inhibitor; 24) BCR-ABL kinase inhibitor; 25) an FGFR inhibitor; 26) CYP11B2 inhibitor; 27) an HDM2 inhibitor, eg an HDM2-p53 interaction inhibitor; 28) a tyrosine kinase inhibitor; 29) cMET inhibitor; 30) a JAK inhibitor; 31) DAC inhibitor; 32) 11b-hydroxylase inhibitor; 33) an IAP inhibitor; 34) a PIM kinase inhibitor; 35) porcupine inhibitor; 36) a BRAF inhibitor, eg V600E BRAF or wild-type BRAF; 37) HER3 inhibitor; 38) MEK inhibitor; or 39) a lipid kinase inhibitor, for example, as described in the present description and in table. 6.

In one embodiment, the second drug is selected from one or more of: Compound A8, Compound A17, Compound A23, Compound A24, Compound A27, Compound A29, Compound A33, and Compound A13.

In other embodiments, the second drug is selected from one or more of: Compound A5, Compound A8, Compound A17, Compound A23, Compound A24, Compound A29, and Compound A40.

In other embodiments, the second drug is selected from one or more of: Compound A9, Compound A16, Compound A17, Compound A21, Compound A22, Compound A25, Compound 28, Compound A48, and Compound 49.

In other embodiments, the second drug is selected from an apoptotic cascade modulator, such as an IDH1 inhibitor or a Bcl-2 or Bcl-XL inhibitor. In one embodiment, the second drug is selected from Compound A21, A14, or combinations thereof. Without being bound by theory, it is known that TIM-3 interacts with PtdSer, which tends to be exposed on the surface of apoptotic cells and can cause immunosuppression. Blockade of the PtdSer-TIM-3 interaction, for example, using an anti-TIM-3 antibody molecule as described herein, may mitigate or overcome immunosuppression.

In other embodiments, the second drug is a CSF-1R inhibitor, such as an anti-CSF-1R antibody or a small molecule inhibitor (such as Compound A15 or A33). These second drugs can inhibit macrophages (eg M2 macrophages). In certain embodiments, such second drugs may facilitate conversion to M1 macrophages.

In embodiments, the second drug is administered at or below a therapeutic dose. In certain embodiments, the concentration of the second drug that is required to achieve inhibition, such as growth inhibition, is lower when the second drug is administered in combination with an anti-TIM-3 antibody molecule than when the second drug is administered alone. In certain embodiments, the concentration of the anti-TIM-3 antibody molecule that is required to achieve inhibition, e.g., growth inhibition, is lower when the anti-TIM-3 antibody molecule is administered in combination with a second drug than when the anti-TIM-3 antibody molecule is administered. individually. In certain combination therapy embodiments, the concentration of the second drug that is required to achieve inhibition, e.g., growth inhibition, is lower than the therapeutic dose of the second drug as monotherapy, e.g., 10-20%, 20-30%, 30 -40%, 40-50%, 50-60%, 60-70%, 70-80% or 80-90% lower. In certain embodiments, in combination therapy, the concentration of the anti-TIM-3 antibody molecule that is required to achieve inhibition, e.g., growth inhibition, is lower than the therapeutic dose of the anti-TIM-3 antibody molecule as monotherapy, e.g., by 10-20%, 20 -30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80% or 80-90% lower.

Detection

In some aspects, the present invention relates to methods for detecting the presence of TIM-3 in a sample, for example, in vitro or in vivo (for example, in a biological sample, for example, in blood, serum, semen or urine, or in a tissue biopsy, for example, from a hyperproliferative or malignancy). The methods described herein can be used to evaluate (eg, monitor treatment or progression, diagnose, and/or stage a disorder described herein, such as an immune disorder, cancer, or infectious disease in an individual). The method may include: (i) contacting a sample (and optionally a reference, such as a control sample), or administering to an individual with an anti-TIM-3 antibody molecule, as described herein, under conditions that allow interaction

- 22 040365 ve, and (ii) detecting whether the formation of a complex between the antibody molecule and the sample (and optionally a reference, for example, a control sample). Complex formation indicates the presence of TIM-3 and may indicate the suitability or need for the treatment described herein. The method may involve, for example, immunohistochemistry, immunocytochemistry, flow cytometry, antibody molecule complexed with magnetic beads, ELISA assays, PCR methods (eg, RT-PCR).

Typically, the anti-TIM-3 antibody molecule used in in vivo and in vitro diagnostic methods is directly or indirectly labeled with a detectable substance to facilitate detection of bound or unbound binding compound. Suitable detectable substances include various biologically active enzymes, prosthetic groups, fluorescent materials, luminescent materials, paramagnetic (eg, nuclear magnetic resonance active) materials, and radioactive materials.

Additional embodiments relate to a method for treating cancer, comprising: identifying in a sample (e.g., a sample from an individual containing cancer cells and optionally immune cells such as TIL) the presence of one, two, or all of PD-L1, CD8, or IFN-γ , thereby providing a value for one, two, or all of PD-L1, CD8, and IFN-γ. In addition, the method may include comparing the values for PD-L1, CD8 and/or IFN-γ with a reference value, such as a control value. If the values for PD-L1, CD8 and/or IFN-γ exceed a reference value, e.g., control values, a therapeutically effective amount of an anti-TIM-3 antibody (e.g., an anti-TIM-3 antibody described herein) is administered to the subject, optionally combination with one or more other agents, thereby treating cancer. The malignant tumor may be, for example, a malignant tumor described herein, such as 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 embodiments, the cancer is ER+ breast cancer or pancreatic cancer.

Also provided is a method for treating cancer, comprising: examining a sample (e.g., a sample of an individual containing malignant cells) for the presence of PD-L1, thereby identifying a value for PD-L1, comparing the value for PD-L1 with a control value, and if the value for PD-L1 is greater than a control value, administering a therapeutically effective amount of an anti-TIM-3 antibody (e.g., an anti-TIM-3 antibody described herein) to the subject, optionally in combination with one or more other agents, for example, an anti-PD-1 antibody molecule thereby carrying out the treatment of a malignant tumor. The cancer may be, for example, a cancer as described herein, such as non-small cell lung adenocarcinoma (NSCLC) (ASA), NSCLC squamous cell carcinoma (SCC), or hepatocellular carcinoma (HCC).

In some aspects, the present invention provides diagnostic or therapeutic kits that include the anti-TIM-3 antibody molecules described herein and instructions for use.

The invention contemplates all combinations of any one or more of the above aspects and/or embodiments, as well as combinations with one or more of the embodiments set forth in the Detailed Description and Examples.

Other features, objectives and advantages of the compositions and methods described in the present description will become clear from the description and drawings and from the claims.

Drawings and tables are attached to this document.

Brief description of the drawings

Each of the drawings is described in the present description in more detail.

In FIG. 1A-1B are exemplary anti-TIM-3 antibodies. In FIG. 1A shows the heavy chain and light chain variable regions of ABTIM3 (SEQ ID NOs: 1 and 2, respectively, in order of occurrence). In FIG. 1B shows the sequence alignment between ABTIM3 variable regions and murine germline antibodies (mouse) (SEQ ID NOS: 134 and 135, respectively, in order of occurrence). CDRs are framed (depicted as white text on a black background in priority documents).

In FIG. 2A-2E illustrate the binding and activity of various anti-TIM-3 antibodies. In FIG. 2A summarizes the affinity data for the mouse ABTIM3 antibody and another TIM-3 binding antibody. In FIG. 2B shows the binding curve for one panel of human TIM-3 antibodies in transfected cells. In FIG. 2C shows the binding curve for a second panel of antibodies, including ABTIM3 (triangles), to human TIM-3 in transfected cells. In FIG. 2D shows the binding curve of ABTIM3 and other anti-TIM-3 antibodies to cynomolgus TIM-3. In FIG. 2E

- 23 040365 shows the affinity of several anti-TIM-3 antibodies for TIM-3 of the cynomolgus monkey. The monoclonal antibody ABTIM3 has the highest affinity among the antibodies tested in these experiments, indicating that it has high cross-reactivity with human and monkey targets.

In FIG. 3A-3B show that anti-TIM-3 monoclonal antibodies, including ABTIM3, bind to the IgV domain, while 4A4 binds to the mucin domain. In FIG. 3A illustrates the recombinant construct used for epitope analysis. In FIG. 3B shows that the anti-TIM-3 monoclonal antibody (anti-TIM-3 antibody #3) and the control anti-PD-L1 monoclonal antibodies (anti-PD-L1 antibodies #1 and #2) bind to the chimeric protein shown in FIG. 3A while anti-TIM-3 #2 and ABTIM3 do not substantially bind to it.

In FIG. 4 illustrates that anti-TIM-3 antibodies anti-TIM-3 antibody #2 and ABTIM3 block TIM-3 binding to PtdSer (phosphatidylserine).

In FIG. 5A-5B illustrate that the anti-TIM-3 antibody ABTIM3 enhances IFN-gamma secretion and proliferation in IL-12 stimulated CD4+ T cells. In FIG. 5A shows the results of a representative experiment where cells were exposed to ABTIM3 antibodies, anti-TIM-3 #2, mIgG1 and control anti-PD-L1 antibodies (from left to right). IFN levels were measured by flow cytometry. In FIG. 5B shows the quantification of IFN-gamma expression in cells exposed to these four antibodies.

In FIG. 6 shows that blockade of ABTIM3 enhances the in vitro cytotoxic activity of purified NK cells.

In FIG. 7 shows that humanized anti-TIM-3 antibodies competed for binding with the mouse parental ABTIM3 antibody in the FACS analysis.

In FIG. 8A-8B illustrate that humanized anti-TIM-3 antibodies bind to cells expressing human TIM-3. In FIG. 8A shows that humanized anti-TIM-3 antibodies bind to cells expressing huTIM-3 in a FACS assay. In FIG. 8B shows that humanized anti-TIM-3 antibodies competed with parental mouse ABTIM3 for huTIM-3 expressing cells in FACS analysis.

In FIG. 9A-9B illustrate the structure of the TIM-3 binding ABTIM3-hum21 Fab. In FIG. 9A shows the general structure of the TIM-3 binding ABTIM3-hum21 Fab. On figa. labeled: 1) identified binding sites for PtdSer, Ca ²⁺ and galectin-9 on human TIM-3 and 2) names of β chains and loops BC, FG and CC'. In FIG. 9B shows a detailed image of ABTIM3hum21 epitope residues on TIM-3 (shown as bars and labeled). In FIG. 9B shows residues 56-61 (GACPVF) as SEQ ID NO: 136 and residues 119-127 (NDEKFNLKL) as SEQ ID NO: 137.

In FIG. 10A-10C compare the ABTIM3-hum21 epitope to the CEACAM-1 binding site on human TIM-3. In FIG. 10A shows a comparison of CEACAM-1 binding residues on TIM-3 (residues 117-120 (IMND) described as SEQ ID NO: 138) (left panel, gray surface, residues labeled) and the ABTIM3-hum21 epitope (right panel, gray surface, residues that overlap with the CECAM1 binding site are indicated). Since TIM-3 is oriented in the same way in both panels, it is clear that the ABTIM3-hum21 epitope overlaps with the CEACAM-1 binding site. In FIG. 10B shows that K122 TIM-3 forms a hydrogen bond with CEACAM1 (left panel) and is completely blocked by ABTIM3-hum21 (right panel). In FIG. 10C shows 2-angle images of the TIM-3/Fab ABTIM3-hum21 and TIM3/CEACAM-1 structures that show a pronounced collision between ABTIM3-hum21 and TIM-3, indicating that ABTIM3-hum21 will disrupt CEACAM- 1 with TIM-3.

In FIG. 11 illustrates a comparison of PtdSer-mediated membrane permeation of mouse TIM-3 (left panel) and ABTIM3-hum21 binding to human TIM-3 (right panel). The two TIM-3 structures are oriented in the same way. The angle of attack of ABTIM3-hum21 is similar to the orientation of the membrane through which TIM-3 penetrates, indicating that ABTIM3-hum21 interferes with PtdSer-mediated entry of TIM-3.

In FIG. 12 shows the cancers with the highest expression of TIM-3 (HAVCR2) from the TCGA database.

In FIG. 13 shows the cancers with the highest expression macrophage expression profile from the TCGA database.

In FIG. 14 shows exemplary cancers having relatively high proportions of patients who are triple positive for PD-L1/CD8/IFN-y.

In FIG. 15 is an exemplary ER+ breast cancer and pancreatic cancer having relatively low proportions of patients that are PDL1/CD8/IFN-y triple positive.

In FIG. 16 shows the proportion of exemplary breast cancer patients who are triple positive for PD-L1/CD8/IFN-y.

In FIG. 17 shows the proportion of patients with illustrative colon cancer who

- 24 040365 are triple positive for PD-L1/CD8/IFN-y.

In FIG. 18 shows peptides monitored in HDx-MS experiments on human TIM-3 (residues 23 to 135 (SEVEYRAEVGQNAYLPCFYTPAAPGNLVPVCWGKGACPVFECGNWLRTDERDVNYWTSRYW

LNGDFRKGDvsLTiENVTLADSGiYCCRiQiPGiMNDEKFNLKLviKPAKVT) in SEQ ID NO: 139). Each bar corresponds to a peptide.

In FIG. Figure 19 illustrates the differences in deuterium uptake by the TIM-3 complex and ABTIM3-hum03 (gray bars) and the TIM-3 and ABTIM3-hum11 complex (black bars) for amino acids 22 to 127. All differences are relative to deuterium uptake by unbound TIM-3 (control ).

In FIG. 20 shows competition between ABTIM3-hum21 and ABTIM3-hum03 and ABTIM3-hum11 for binding to human TIM3 as determined by flow cytometric analysis.

In FIG. 21 is a representative sensogram obtained from a Biacore competitive assay study of competition between 1st antibody and 2nd antibody for immobilized human TIM3.

In FIG. 22 shows that ABTIM3 increases proliferation in a coculture containing dendritic cells and T cells (DC-T coculture). DC-T cocultures were incubated without antibody or with titrated dilution series (0.01-25 μg/ml) of the following antibodies: mouse IgG1 (control), ABTIM3, or anti-TIM3 antibody #3.

In FIG. 23A-23B show the concentration of ABTIM3-hum11 found in rodent serum over time. The indicated dosages were injected into mice or rats and the concentration of the antibody in the blood at the indicated time points was calculated. In FIG. 23A shows the mean serum concentration of BTIM3-hum11 in mice after antibody challenge. In FIG. 23B shows the mean serum concentration of ABTIM3-hum11 in rats after antibody challenge.

Brief description of the tables

Each of the tables is described in the present description in more detail.

In table. 1 summarizes the sequences of the mouse anti-TIM-3 antibody ABTIM3.

In table. 2 shows the amino acid sequences of the heavy chain variable domain and the light chain variable domain of ABTIM3.

In table. 3 shows the amino acid sequences of the heavy chain CDR and the light chain CDR of ABTIM3.

In table. 4 summarizes the amino acid and nucleotide sequences for mouse anti-TIM-3 antibody molecules and humanized anti-TIM-3 antibody molecules. Antibody molecules include mouse ABTIM3 and humanized anti-TIM-3 antibodies: ABTIM3-hum01, ABTIM3hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3 -hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3hum20, ABTIM3-hum21, ABTIM3-hum22, and ABTIM3-hum23. This table lists the amino acid and nucleotide sequences of the heavy and light chain CDRs, the amino acid and nucleotide sequences of the heavy and light chain variable regions, and the amino acid and nucleotide sequences of the heavy and light chains.

In table. 5 shows the amino acid sequences of the human IgG heavy chain and human kappa light chain constant regions.

In table. 6 summarizes individual drugs that can be administered in combination with anti-TIM-3 antibody molecules and other immunomodulators (e.g., one or more of a co-stimulatory molecule activator and/or immune response checkpoint molecule inhibitor) described herein. In table. 6 shows from left to right the following: the compound designation for the second drug, the structure of the compound, and the patent publication(s) in which the compound is described.

In table. 7 summarizes the K _D values for anti-TIM-3 antibody binding to activated PBMCs.

In table. 8 summarizes the K _D values for anti-TIM-3 antibody binding to the PD-L1 IgV/TIM-3 mucin construct.

In table. 9 summarizes the K _D values for a panel of humanized anti-TIM-3 antibodies as measured by the Biacore assay.

In table. 10 summarizes the K _D values for anti-TIM-3 antibody binding to cells expressing human TIM-3.

In table. 11 summarizes the K _D values for anti-TIM-3 antibody binding to TIM-3Ig.

In table. 12 summarizes the amino acid sequences used to determine the crystal structure.

In table. 13 summarizes the amino acids in TIM-3 and anti-TIM-3 antibody that are involved in the binding interaction.

- 25 040365

In table. 14 summarizes the Biacore competitive assay runs.

In table. 15 summarizes the results of the Biacore competitive analysis.

In table. 16 summarizes the pharmacokinetic properties of ABTIM3-hum11.

Detailed description

T-cell immunoglobulin and mucin domain 3 (TIM-3, also known as hepatitis A virus cell receptor 2 and HAVCR2) is a cell surface protein expressed, for example, on activated CD4+ and CD8+ T cells, natural regulatory T cells (nTreg ), NK cells, and innate cells such as macrophages, monocytes, and dendritic cells (DC). TIM-3 is generally not expressed on naïve T cells, but rather is upregulated on activated effector T cells, such as the PD-1+ subset of cells. TIM-3 is also expressed in tissue natural regulatory cells and in mouse models. TIM3+ Tregs have been shown to have a more suppressive phenotype, while TIM-3+ Tregs have also been shown to correlate with disease severity in NSCLC, hepatic cell carcinoma, and ovarian carcinoma. TIM-3 is constitutively expressed on DCs, monocytes/macrophages, and NK cells, and blockade of TIM-3 has been shown to correlate with increased cytotoxicity in NK cells; increased secretion of IL-12/TNF-a by monocytes/macrophages; and increased expression of NF-kB in DC. Blockade of TIM-3 (partially alone and additively or synergistically in combination with blockade of the PD-1 cascade) has shown antitumor efficacy in several preclinical cancer models, including CT26 colon carcinoma (Sakuishi et al., J Exp Med. 2010;207 (10 ):2187-94), WT3 sarcoma, and TRAMP-C1 prostate carcinoma (Ngiow et al., Cancer Res. 2011; 71(10):3540-3551). Recent studies have demonstrated that TIM-3 is an important participant in the depletion and suppression of effector T cells that occur in chronic immune conditions such as infection, such as bacterial or viral, and malignancy, both in humans and in experimental models. TIM-3 has been described as an inhibitory receptor at immunological synapses, and blocking TIM-3 may enhance the immune response against infection and cancer.

Blockade of TIM-3 has been shown to restore activity in effector cells such as cytokine secretion and proliferation. In virus-depleted cell populations, for example, HCV-infected cells expressing TIM-3 (TIM3+ cells) express less TNF-alpha and IFN-gamma cytokines than TIM-3 negative cells in both populations of effector cells: CD4+ and CD8+ T cells (Golden-Mason et al., 2009, J. Virol, 83:9122). Blockade of TIM-3 restores proliferation in CD8+ T cells from an HIV patient or in cells that reproduce viral depletion (Jones et al., 2008, J. Exp. Med., 205:2763), or proliferation and secretion of IFN-γ and/or TNF-α in NY-ESO-1 specific T cells from PBMCs from metastatic patients (Fourcade et al., 2010, J. Exp. Med., 207:2175).

Blockade of TIM-3 can also reduce the suppressor activity of regulatory T cells. TIM-3+ T cells have been found to concentrate in tumors and contribute to the immunosuppressive environment of the tumor (Sakuishi et al., 2013, Oncoimmunology, 2:e23849; Gao et al., 2012, Plos One; and Yan et al. , 2013, Plos One.). Thus, blockade of TIM-3, for example, by antibodies that inhibit TIM-3 function, can enhance the immune response against infection and antitumor immunity.

TIM-3 is also involved in the regulation of the immune response through the activity of macrophages. Blockade of TIM-3 leads to an increase in TLR-mediated production of IL-12 (Zhang et al., 2010, J Leukoc Biol, 91:189). Thus, blockade of TIM-3 can increase the immunostimulatory properties of macrophages, enhancing the immune response against infection and antitumor activity.

TIM-3 has five described ligands: galectin-9 (Gal-9), phosphatidylserine (PtdSer), HMGB1, semaphorin-4A, and CEACAM-1. The S-type lectin galectin-9 can inhibit TIM-3-associated Th1 effector function and induce apoptosis on TIM-3-expressing T cells in a mouse model. PtdSer is normally located on the intracellular side of the plasma membrane, but moves to the extracellular side during apoptosis. PtdSer binds the cleft that is preserved in all three members of the human TIM family (TIM-1, 3, 4). Inhibition of PtdSer binding to TIM-3 can activate the T cell response. Galectin-9 is secreted by tumor cells and may contribute to escaping antitumor immunity. The HMGB1 DNA alarmin, for which TIM3 can act as a sinker, may interfere with HMGB1/RAGE interactions that stimulate innate immunity. Semaphorin-4A and CEACAM-1 (another immune response checkpoint molecule whose inhibition can enhance the immune response) can interact with TIM-3 both in the cis configuration as a heterodimer on T cells and in the trans configuration as a ligand. The interaction between CEACAM-1 and TIM-3 may help mediate blocking of signal transduction in the immune response. Co-blockade of TIM-3 and CEACAM-1 in CT26 colon carcinoma showed similar efficacy to that observed for co-blockade of PD-L1 and TIM-3.

The cytoplasmic tail of TIM-3 has seven sites for tyrosine phosphorylation and

- 26 040365 has no known inhibitory (ie ITIM) motifs, indicating that TIM-3 could costimulate with the T cell receptor, causing functional depletion through increased T cell signaling. TIM-3 can interact with Fyn and promote the accumulation of CD148 and CD45 receptor phosphatases in the immunological synapse. The presence of CEACAM-1 as a co-receptor in the TIM-3/CEACAM-I heterodimer indicates that this co-expression may result in inhibitory signaling in T cells via the ITIM motif in the cytoplasmic tail of CEACAM-1, which has been shown to interacts with both SHP1 and SHP2.

The present disclosure describes antibody molecules that bind to TIM-3 with high affinity and specificity. In one embodiment, humanized anti-TIM-3 antibodies are described. Additional aspects of the invention include nucleic acid molecules encoding antibody molecules, expression vectors, host cells, and methods for producing antibody molecules are also provided. Also contemplated are immunoconjugates, polyspecific or bispecific antibody molecules, and pharmaceutical compositions containing antibody molecules. The anti-TIM-3 antibody molecules described herein can be used (alone or in combination with other agents or therapeutic regimens) for the treatment, prevention and/or diagnosis of immune disorders, cancer, infectious disease, Crohn's disease, sepsis, SIRS ( systemic inflammatory response syndrome) and glomerulonephritis. Thus, compositions and methods for detecting TIM-3 as well as methods for treating various disorders, including cancer and immune disorders, are described herein using anti-TIM-3 antibody molecules.

The term TIM-3 includes isoforms, mammalian, eg human, TIM-3, human TIM-3 species homologues, and analogues containing at least one epitope in common with TIM-3. The amino acid sequence of TIM-3, eg human TIM-3, is known in the art, eg Sabatos et al., 2003. Nat Immunol, 4(11):1102.

Definitions

As used herein, the singular form refers to one or more than one (eg, at least one) grammatical entity in the singular form.

The term or is used herein to refer to and is used interchangeably with the term and/or unless the context clearly indicates otherwise.

Approximately and approximately generally means an acceptable degree of error for the quantity measured, given the nature or accuracy of the measurements. Exemplary error rates are within 20 percent (%), typically within 10%, and more specifically within 5% of a given value or range of values.

The compositions and methods described herein encompass polypeptides and nucleic acids having said sequences, or sequences substantially identical or similar thereto, such as sequences that are at least 85%, 90%, 95% or greater identical to said sequence. In the context of amino acid sequence, the term substantially identical is used herein to refer to the first amino acid that contains a sufficient or minimum number of amino acid residues that i) are identical or ii) are conservative substitutions for aligned amino acid residues in the second amino acid sequence such that the first and second amino acid sequences may share a common structural domain and/or a common functional activity. Examples are amino acid sequences that contain a common structural domain having at least about 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with reference sequence, such as the sequence described in the present description.

In the context of a nucleotide sequence, the term substantially identical is used herein to refer to a first nucleic acid sequence that contains a sufficient or minimum number of nucleotides that are identical to aligned nucleotides in a second nucleic acid sequence such that the first and second nucleotide sequences encode polypeptides having a common functional activity, or encode the general structural polypeptide domain or the general functional activity of the polypeptide. Examples are nucleotide sequences having at least about 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with a reference sequence, e.g. described in the present description.

The term functional variant refers to polypeptides that have a substantially identical amino acid sequence to a naturally occurring sequence, or are encoded by a substantially identical nucleotide sequence, and are capable of having one or more of the activities of a naturally occurring sequence.

To determine the percent identity of two amino acid sequences or two nucleic acid sequences, the sequences are aligned for optimal

- 27 040365 comparisons (for example, gaps can be made in one or both of the first and second amino acid or nucleic acid sequences for optimal alignment, and non-homologous sequences can be neglected for comparison purposes). In a preferred embodiment, the length of the reference sequence aligned for comparison purposes is at least 30%, such as at least 40%, 50%, 60%, such as at least 70%, 80%, 90%, 100% of the length reference sequence. The amino acid residues or nucleotides at the respective amino acid positions or nucleotide positions are then compared. When a 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 at that position.

Percent identity between two sequences is a function of the number of identical positions between sequences, taking into account the number of gaps and the length of each gap that must be made for optimal alignment of the two sequences.

Sequence comparison and determination of percent identity between two sequences can be performed using a mathematical algorithm. In some embodiments, percent identity between two amino acid sequences is determined using the algorithm of Needleman and Wunsch ((1970) J. Mol. Biol. 48:444-453), which was included in the GAP program of the GCG software package (available at http:// www.gcg.com), using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a gap extension weight of 1, 2, 3, 4, 5, or 6. In certain embodiments, percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package (available at http://www.gcg.com) using the NWSgapdna matrix. CMP and a skip weight of 40, 50, 60, 70, or 80 and a skip continuation weight of 1, 2, 3, 4, 5, or 6. One suitable parameter set (and the parameter set that should be used unless otherwise indicated) is the estimated a Blossum 62 matrix with a gap penalty of 12, a gap continuation penalty of 4, and a frameshift gap penalty of 5.

Percent identity between two amino acid or nucleotide sequences can be determined using the algorithm of E. Meyers and W. Miller ((1989) CABIOS, 4:11-17), which is included in the ALIGN program (version 2.0), using the PAM120 residue weight table, gap continuation penalty 12 and gap penalty 4.

The nucleic acid and protein sequences described herein can be used as a query sequence to conduct searches against public databases, for example, to identify other members of a family or related sequences. Such searches can be performed using the NBLAST and XBLAST (version 2.0) programs from Altschul, et al. (1990) J. Mol. Biol. 215:403-10. Nucleotide BLAST searches can be performed using the NBLAST program, weight=100, wordlength=12, to obtain nucleotide sequences homologous to a nucleic acid as described herein. BLAST protein searches can be performed using the XBLAST program, weight=50, wordlength=3, to obtain amino acid sequences homologous to the protein molecules described herein. To perform gapped alignments for comparison purposes, Gapped BLAST can be used 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 settings of the respective programs (eg XBLAST and NBLAST). See www.ncbi.nlm.nih.gov.

As used herein, the term hybridize under low stringency, moderate stringency, high stringency, or very high stringency describes the conditions for hybridization and washing. Guidelines for conducting hybridization reactions can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6, which is incorporated by reference. This reference describes aqueous and non-aqueous methods, and any of them can be used. Specific hybridization conditions referred to herein are as follows: 1) low stringency hybridization conditions in 6X sodium chloride/sodium citrate (SSC) at approximately 45°C followed by two washes in 0.2X SSC, 0.1% SDS at least 50°C (washing temperature can be increased up to 55°C for low stringency conditions); 2) moderate stringency hybridization conditions in 6X SSC at approximately 45°C followed by one or more washes in 0.2X SSC, 0.1% SDS at 60°C; 3) high stringency hybridization conditions in 6X SSC at approximately 45°C followed by one or more washes in 0.2X SSC, 0.1% SDS at 65°C; and preferably 4) very stringent hybridization conditions are 0.5M sodium phosphate, 7% SDS at 65°C followed by one or more washes in 0.2X SSC, 1% SDS at 65°C. Very severe conditions (4) are suitable conditions and conditions should be used unless otherwise indicated.

It is understood that the molecules described herein may have additional conservative amino acid substitutions or non-essential amino acid substitutions that do not have a significant effect on their function.

- 28 040365

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 having similar side chains are defined in the art. These families include amino acids with 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 (eg, alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (eg, threonine, valine, isoleucine), and aromatic side chains (eg, tyrosine, phenylalanine , tryptophan, histidine).

The terms polypeptide, peptide, and protein (if single chain) are used interchangeably herein.

The terms nucleic acid, nucleic acid sequence, nucleotide sequence or polynucleotide sequence and polynucleotide are used interchangeably.

The term isolated, as used herein, refers to material that has been removed from its original or natural environment (eg, the natural environment if it is naturally occurring). 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 its coexisting materials in the natural system is isolated. Such polynucleotides may be part of a vector and/or such polynucleotides or polypeptides may be part of a composition and still be isolated because such vector or composition is not part of the environment in which it occurs naturally.

Various aspects of the compositions and methods described herein are described in more detail below. Throughout the description, additional definitions are provided.

Antibody molecules

In some embodiments, the antibody molecule binds to TIM-3 of a mammal, such as a human. For example, an antibody molecule binds specifically to an epitope, such as a linear or conformational epitope (eg, an epitope as described herein) on TIM-3. In some embodiments, the epitope is at least a portion of a human or cynomolgus TIM-3 IgV domain.

As used herein, the term antibody molecule refers to a protein, such as an immunoglobulin chain or fragment thereof, containing at least one immunoglobulin variable domain sequence. The term antibody molecule includes, for example, a monoclonal antibody (including a full-length antibody that has an immunoglobulin Fc region). In one embodiment, the antibody molecule comprises a full length antibody or a full length immunoglobulin chain. In one embodiment, the antibody molecule comprises an antigen-binding or functional fragment of a full-length antibody or a full-length immunoglobulin chain.

In one embodiment, the antibody molecule is a monospecific antibody molecule and binds a single epitope. An example is a monospecific antibody molecule having multiple immunoglobulin variable domain sequences, each of which binds the same or substantially the same epitope.

In one embodiment, the antibody molecule is a multispecific antibody molecule, e.g., it comprises a plurality of immunoglobulin variable domain sequences, wherein a first immunoglobulin variable domain sequence of the plurality has binding specificity for a first epitope and a second immunoglobulin variable domain sequence of the plurality has binding specificity for second epitope. In one embodiment, the first and second epitopes are on the same antigen, eg, on the same protein (or subunit of a multimeric protein). In one embodiment, the first and second epitopes overlap or substantially overlap. In one embodiment, the first and second epitopes do not overlap or do not substantially overlap. In one embodiment, the first and second epitopes are on different antigens, such as on different proteins (or different subunits of a multimeric protein). In one embodiment, the multispecific antibody molecule contains a third, fourth, or fifth immunoglobulin variable domain. In one embodiment, the multispecific antibody molecule is a bispecific antibody molecule, a trispecific antibody molecule, or a tetraspecific antibody molecule.

In one embodiment, the multispecific antibody molecule is a bispecific antibody molecule. A bispecific antibody molecule has specificity for no more than two antigens. A bispecific antibody molecule is characterized by the first sequence of the immunoglobulin variable domain, which has binding specificity.

- 29 040365 for the first epitope, and a second immunoglobulin variable domain sequence that has binding specificity for the second epitope. In one embodiment, the first and second epitopes are on the same antigen, eg on the same protein (or subunit of a multimeric protein). In one embodiment, the first and second epitopes overlap or substantially overlap. In one embodiment, the first and second epitopes do not overlap or do not substantially overlap. In one embodiment, the first and second epitopes are on different antigens, such as different proteins (or different subunits of a multimeric protein). In one embodiment, the bispecific antibody molecule comprises a light chain variable domain sequence and a light chain variable domain sequence that have binding specificity for a first epitope, and a heavy chain variable domain sequence and a light chain variable domain sequence that have binding specificity for a second epitope. . In one embodiment, the bispecific antibody molecule comprises a half-antibody having binding specificity for a first epitope and a half-antibody having binding specificity for a second epitope. In one embodiment, the bispecific antibody molecule comprises a half-antibody or fragment thereof having a binding specificity for a first epitope and a half-antibody or fragment thereof having a binding specificity for a second epitope. In one embodiment, the bispecific antibody molecule comprises an scFv or fragment thereof that has a binding specificity for a first epitope and an scFv or fragment thereof that has a binding specificity for a second epitope. In one embodiment, the first epitope is located on TIM-3 and the second epitope is located on PD-1, LAG-3, CEACAM (e.g., CEACAM-1, CEACAM-3 and/or CEACAM-5), PD-L1 or PD-L2 .

In one embodiment, the antibody molecule includes a diantibody, a single chain molecule, as well as an antigen-binding fragment of an antibody (eg, Fab, F(ab') ₂ and Fv). For example, an antibody molecule may include a heavy (H) chain variable domain sequence (abbreviated herein as VH) and a light chain (L) variable domain sequence (abbreviated herein as VL). In one embodiment, the antibody molecule contains or consists of a heavy chain or a light chain (abbreviated herein as a half antibody). In another example, 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, single chain antibodies (e.g., scFv), single variable domain antibodies, diabodies (Dab) (bivalent and bispecific), and chimeric (e.g., humanized) antibodies, which can be obtained by modifying whole antibodies or antibodies synthesized de novo using recombinant DNA technologies. These functional antibody fragments retain the ability to selectively bind to their respective antigen or receptor. Antibodies and antibody fragments can be from any class of antibodies, including but not limited to IgG, IgA, IgM, IgD, and IgE, and from any subclass (eg, IgG1, IgG2, IgG3, and IgG4) of antibodies. The preparation of antibody molecules can be monoclonal or polyclonal. The antibody molecule can also be a human antibody, a humanized antibody, a CDR-grafted antibody, or an in vitro generated antibody. The antibody may have a heavy chain constant region selected from, for example, IgG1, IgG2, IgG3, or IgG4. The antibody may have a light chain selected from, for example, kappa or lambda. The term immunoglobulin (Ig) is used herein interchangeably with the term antibody.

Examples of antigen-binding fragments of an antibody molecule include: (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL, and CH1 domains; (ii) F (ab') ₂ fragment, divalent fragment containing two Fab-fragment connected by a disulfide bridge in the hinge region; (iii) an Fd fragment consisting of VH and CH1 domains; (iv) an Fv fragment consisting of the VL and VH domains of one arm of an antibody, (v) a diaantibody fragment (dAb) which consists of a VH domain; (vi) a camelid variable domain or camelized variable domain; (vii) single chain Fv (scFv), see, for example, Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. sci. USA 85:5879-5883); (viii) single domain antibody. These antibody fragments can be obtained using any suitable method, including several conventional methods known to those skilled in the art, and the fragments can be screened for utility in the same way as intact antibodies.

The term antibody includes intact molecules as well as their functional fragments. The constant regions of an antibody can be altered, eg, by mutation, to modify the properties of the antibody (eg, to increase or decrease one or more of: Fc receptor binding, antibody glycosylation, cysteine residue count, cellular effector function, or complement function).

The antibodies described herein may also be single domain anti

- 30 040365 bodies. Single domain antibodies may include antibodies whose complementarity determining regions are part of a single domain polypeptide. Examples include, but are not limited to, heavy chain antibodies, antibodies naturally lacking light chains, single domain antibodies derived from conventional 4 chain antibodies, engineered antibodies, and single domain scaffolds other than those derived from antibodies. Single domain antibodies can be any prior art single domain antibody or any future single domain antibody. Single domain antibodies can be derived from any species, including, but not limited to, mouse, human, camel, llama, fish, goat, rabbit, and ruminants. In some aspects, a single domain antibody is a naturally occurring single domain antibody known as a heavy chain antibody lacking light chains. Such single domain antibodies are described, for example, in WO 9404678. For clarity, this heavy chain antibody derived variable domain naturally lacking a light chain is known herein as VHH or nanoantibody to distinguish it from the conventional VH of heavy chain immunoglobulins. . Such a VHH molecule may be derived from antibodies induced in Camelidae species such as camel, llama, dromedary, alpaca and guanaco. Species other than the Camelidae can produce heavy chain antibodies naturally lacking a light chain; such VHHs are also provided.

The VH and VL regions can be subdivided into regions of hypervariability called complementarity determining regions (CDRs) located between regions that are more conserved regions called framework regions (FRs). The extent of the framework region and CDR has been accurately determined by a number of methods (see, Kabat, E.A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242; Chothia, C. et al (1987) J. Mol Biol 196:901-917 and the AbM definition used by Oxford Molecular's AbM antibody modulation software See mainly for example Protein Sequence and Structure Analysis of Antibody Variable Domains Antibody Engineering Lab Manual (Ed.: Duebel, S. and Kontermann, R., Springer-Verlag, Heidelberg) In some embodiments, the following definitions are used: definition of heavy chain variable domain CDR1 according to AbM and definitions of other CDRs according to Kabat.In certain embodiments, Kabat definitions are used for all CDRs.In addition, embodiments described in relation to CDRs according to Kabat or AbM can also be implemented using hypervariable loops of Chothia. Each VH and VL typically includes three CDRs and four FRs, arranged from N-terminus to C-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

As used herein, an immunoglobulin variable domain sequence refers to an amino acid sequence that can form 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- or C-terminal amino acids, or may include other changes that are compatible with the formation of a protein structure.

The term antigen binding site refers to the portion of an antibody molecule that contains determinants that form a contact surface that binds to a TIM-3 polypeptide or epitope thereof. With respect to proteins (or protein mimetics), the antigen binding site typically includes one or more loops (at least, for example, four amino acids or amino acid mimetics) that form a contact surface that binds to the TIM-3 polypeptide. Typically, the antigen-binding site of an antibody molecule includes at least one or two CDRs, or more particularly at least three, four, five, or six CDRs.

The terms compete or cross-compete are used interchangeably herein to refer to the ability of an antibody molecule to prevent an anti-TIM-3 antibody molecule, such as the anti-TIM-3 antibody molecule described herein, from binding to a target, such as human TIM-3. Interfering with binding may be direct or indirect (eg, through allosteric modulation of the antibody or target molecule). The degree to which an antibody molecule is able to prevent another antibody molecule from binding to a target, and thus whether competition can be asserted, can be determined using a competitive binding assay, such as a FACS, ELISA, or BIACORE assay. In some embodiments, the competitive binding assay is a quantitative competitive assay. In some embodiments, the first anti-TIM-3 antibody molecule is said to compete for target binding with the second anti-TIM-3 antibody molecule when the binding of the first antibody molecule to the target is reduced by 10% or more, e.g., 20% or more, 30% or more, 40% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, 95% or more, 98% or more, 99% or more in a competitive binding assay (eg, a competitive assay described herein).

As used herein, the term epitope refers to portions of an antigen (eg, human TIM-3) that specifically interact with an antibody molecule. Such parts referred to herein as epitope determinants typically contain or are part of elements such as amino acid side chains or sugar side chains. The epitope determinant can be determined by methods known in the art or described herein, for example, by crystallography or hydrogen-deuterium exchange. At least one or some of the moieties on an antibody molecule that interact specifically with an epitope determinant are typically located in a CDR (multiple CDRs). As a rule, an epitope has certain characteristics of a three-dimensional structure. Typically, an epitope has certain charge characteristics. Some epitopes are linear epitopes while others are conformational epitopes.

In one embodiment, the epitope determinant is a portion on an antigen, such as an amino acid side chain or a sugar side chain, or a portion thereof, which, when the antigen and antibody molecule co-crystallize, is at a predetermined distance, such as within 5 angstroms, from the portion on an antibody molecule, referred to herein as a crystallographic epitope determinant.

The crystallographic epitope determinants of an epitope are collectively referred to herein as a crystallographic epitope.

The first antibody molecule binds to the same epitope as the second antibody molecule (e.g., a reference antibody molecule, e.g., an antibody molecule described herein, e.g., ABTIM3-hum21, ABTIM-hum11, or ABTIM3-hum03) if the first antibody is specific interacts with the same epitope determinants on the antigen as the second or reference antibody, for example, when the interaction is measured in the same way for both the antibody and the second or reference antibody. Epitopes that overlap share at least one epitope determinant in common. The first antibody molecule binds an overlapping epitope to a second antibody molecule (e.g., a reference antibody molecule, e.g., an antibody described herein, e.g., ABTIM3-hum21, ABTIM-hum11, or ABTIM3-hum03) when both antibody molecules specifically interact with a common epitope determinant. The first and second antibody molecules (e.g., a reference antibody molecule, e.g., an antibody molecule described herein, e.g., ABTIM3-hum21, ABTIM-hum11, or ABTIM3-hum03) bind substantially overlapping epitopes if at least half of the epitope determinants of the second or reference antibodies occur as epitope determinants in the epitope of the first antibody. The first and second antibody molecules (e.g., a reference antibody molecule, e.g., an antibody molecule described herein, e.g., ABTIM3-hum21, ABTIM-hum11, or ABTIM3-hum03) bind substantially the same epitope if the first antibody molecule binds at least half of the central epitope determinants of the second or reference antibody, where the central epitope determinants are determined by crystallography and hydrogen-deuterium exchange, for example, including residues Val24, Glu25, Thr41, Glu121, Lys122, Phe123, Asn124, Leu125, Lys126, Leu127, Val128 , Gly56, Ala57, Cys58, Pro59, Val60 and Phe61 human TIM-3.

The terms monoclonal antibody or monoclonal antibody composition, as used herein, refers to a preparation of antibody molecules with a single molecular composition. A monoclonal antibody composition exhibits one binding specificity and affinity for a particular epitope. The monoclonal antibody can be produced by hybridoma technology or by methods that do not use hybridoma technology (eg, recombinant methods).

An effective human protein is one that does not induce a neutralizing antibody response, such as a human anti-mouse (HAMA) response. HAMA can be a problem in some cases, for example if the antibody molecule is administered multiple times, such as in the treatment of a chronic or relapsing disease. The HAMA response may render multiple antibody administrations ineffective due to increased serum clearance of antibodies (see, e.g., Saleh et al., Cancer Immunol. Immunother., 32:180-190 (1990)) and also due to potential allergic reactions (see, for example, LoBuglio et al., Hybridoma, 5:5117-5123 (1986)).

The antibody molecule may be a polyclonal or monoclonal antibody. In other embodiments, the antibody may be recombinantly produced, eg produced by any suitable phage display or combinatorial methods.

Various phage display methods and combinatorial methods for producing antibodies are known in the art (as described, for example, in Ladner et al., US Patent No. 5,223,409; Kang et al. No. WO 91/17271 Winter et al. International Publication No. WO 92/20791 Markland et al. International Publication No. WO 92/15679 Breitling et al. International Publication No. WO93/01288 McCafferty et al. Publication No. WO 92/01047 Garrard et al., International Publication No. WO 92/09690, Ladner et al., International Publication No. WO 90/02809, 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 352:624-32 040365

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 88:7978-7982, the contents of which are incorporated herein by reference).

In some embodiments, the antibody is a fully human antibody (e.g., an antibody produced by a mouse that has been genetically modified to produce antibodies with human immunoglobulin sequences) or a non-human antibody, such as a rodent (mouse or rat), goat, primate (e.g. , monkey), camel. In certain embodiments, the non-human antibody is a rodent (mouse or rat antibody). Methods for producing rodent antibodies are known in the art.

Human monoclonal antibodies can be generated using transgenic mice containing human immunoglobulin genes instead of the mouse system. Splenocytes from these transgenic mice immunized with an antigen of interest are used to generate hybridomas that secrete a human mAb with a specific affinity for epitopes from a human protein (see, for example, Wood et al., WO 91/00906, Kucherlapati et al. ., PCT Publication WO 91/10741; Lonberg et al., International Application WO 92/03918; Kay et al., International Application 92/03917; Lonberg, N. et al. 1994 Nature 368:856-859; Green, L.L. et al 1994 Nature Genet 7:13-21; Morrison, S. L. 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 90:3720-37'24; Bruggeman et al 1991 Eur J Immunol 21:1323-1326).

The antibody may be an antibody in which the variable region or portion thereof, such as a CDR, is produced in a non-human body, such as a rat or mouse. Also contemplated are chimeric antibodies, CDR-grafted antibodies, and humanized antibodies. Also contemplated are antibodies produced in a non-human, eg rat or mouse, and then modified, eg in a variable framework region or constant region, to reduce antigenicity in humans.

Chimeric antibodies can be produced by any suitable recombinant DNA method. Several such methods are known in the art (see Robinson et al., International Patent Publication PCT/US86/02269; Akira, et al., European Patent Application 184187; Taniguchi, M., European Patent Application 171496; Morrison et al., European Patent Application 173494 Neuberger et al., WO 86/01533 Cabilly et al., US Patent No. 4816567 Cabilly et al., European Patent Application 125023 Better et al. ; Liu et al. (1987) PNAS 84:3439-3443; Liu et al., 1987, J. Immunol. 139:3521-3526; Sun et 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).

In a humanized or CDR-grafted antibody, at least one or two, but typically all three, of the recipient CDR (Immunoglobulin heavy and/or light chain) are replaced by a donor CDR. In an antibody, at least a portion of the non-human CDR may be replaced, or only some of the CDRs may be replaced with non-human CDRs. It is only necessary to change the set of CDRs required to bind the humanized antibody to TIM-3. In some embodiments, the donor is a rodent antibody, such as a rat or mouse antibody, and the recipient is a human framework or human consensus framework. Typically, an immunoglobulin providing a CDR is referred to as a donor and an immunoglobulin providing a framework region is referred to as an acceptor. In some embodiments, the implementation of the donor immunoglobulin is not human (eg, rodent). An acceptor framework is typically a naturally occurring (eg, human) framework or a consensus framework or sequence about 85% or more, eg, 90%, 95%, 99% or more identical thereto.

As used herein, the term consensus sequence 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 proteins, each position in the consensus sequence is occupied by the amino acid that occurs most frequently at that position in the family.If two amino acids occur with equal frequency, either of them can be included in the consensus sequence.The consensus framework refers to the framework region in the consensus sequence of an immunoglobulin.

An antibody can be humanized by any suitable method, and several such methods are known in the art (see, for example, Morrison, S. L., 1985, Science 229:1202-1207, by Oi et al., 1986, BioTechniques 4:214, and Queen et al US 5585089, US 5693761 and US 5693762, the contents of which are incorporated herein by reference).

Humanized or CDR-grafted antibodies can be produced by CDR grafting or CDR replacement, where one, two or all of the immunoglobulin chain CDRs can be replaced. See, for example, US Pat. No. 5,225,539; 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 US 5225539, contents of each cat

- 33 040365 ryh expressly included in the present description by reference. Winter describes a CDR transplant method that can be used to produce humanized antibodies (GB Patent Application GB 2188638A, filed March 26, 1987; Winter, US 5225539, the contents of each of which are expressly incorporated herein by reference).

Also contemplated are humanized antibodies in which specific amino acids are substituted, deleted, or inserted. Donor amino acid selection criteria are described, for example, in US Pat. No. 5,585,089, for example in columns 12-16 of US Pat. No. 5,585,089, the contents of which are incorporated herein by reference.

Other methods for humanizing antibodies are described in Padlan et al. EP 519596A1 published December 23, 1992.

The antibody molecule may be a single chain antibody. A single chain antibody (scFV) can be engineered (see, for example, Colcher, D. et al. (1999) All N Y Acad Sci 880:263-80; and Reiter, Y. (1996) Clin Cancer Res 2:245 -52). A single chain antibody can be dimerized or multimerized to produce polyvalent antibodies having specificity for different epitopes on the same target protein.

In some embodiments, the antibody molecule has a heavy chain constant region selected from, for example, IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE heavy chain constant regions; in particular selected, for example, from heavy chain (eg human) constant regions of IgG1, IgG2, IgG3 and IgG4. In another embodiment, the antibody molecule has a light chain constant region selected, for example, from kappa and lambda (eg, human) light chain constant regions. The constant region can be changed, for example, by mutation, to modify the properties of the antibody (for example, to increase or decrease one or more of: Fc receptor binding, antibody glycosylation, number of cysteine residues, effector cell function, and/or complement function). In some embodiments, the antibody has an effector function and can fix complement. In other embodiments, the antibody does not attract effector cells or does not fix complement. In certain embodiments, the antibody has reduced or no ability to bind the Fc receptor. For example, it may be an isotype or subtype, fragment, or other mutant that does not support binding to the Fc receptor, for example, it has a mutated or deleted Fc receptor binding site.

In some embodiments, the implementation of the constant region of the antibody is changed. Methods for changing the constant region of an antibody are known in the art. Antibodies with an altered function, such as an altered affinity for an effector ligand, such as an FcR on a cell, or the C1 component of complement, can be obtained by replacing at least one amino acid residue in the constant portion of the antibody with another residue (see, for example, EP 388151A1, patent US No. 5624821 and US patent No. 5648260, the contents of all of which are incorporated into the present description by reference). Also contemplated are amino acid mutations that stabilize the antibody structure, such as S228P (EU nomenclature, Kabat nomenclature S241P) in human IgG4. A similar type of change can be described which, when applied to an immunoglobulin of a mouse or other species, will reduce or abolish these functions.

In some embodiments, the amino acids in an anti-TIM-3 antibody molecule are only canonical amino acids. In some embodiments, the anti-TIM-3 antibody molecule contains naturally occurring amino acids; their analogues, derivatives and related compounds; amino acid analogs having different side chains; and/or all stereoisomers of any of the above. An anti-TIM-antibody molecule may contain D- or L-optical isomers of amino acids and peptidomimetics.

The polypeptide of an anti-TIM-3 antibody molecule may be linear or branched, it may contain modified amino acids, and it may be interrupted by non-amino acids. The antibody molecule can also be modified, for example, by disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation such as conjugation to a labeling moiety. The polypeptide may be isolated from natural sources, may be produced by recombinant methods from a eukaryotic or prokaryotic host, or it may be the product of synthetic procedures.

An antibody molecule may be derivatized or linked to another functional molecule (eg, another peptide or protein). As used herein, a derivatized antibody molecule is one that has been modified. Derivatization methods include, but are not limited to, attachment of a fluorescent moiety, a radionucleotide, a toxin, an enzyme, or an affinity ligand such as biotin. Thus, antibody molecules are meant to include derivatized or otherwise modified forms of the antibodies described herein, including immunoadhesion molecules. For example, an antibody molecule may be operably linked (via chemical bonding, genetic fusion, non-covalent association, or otherwise) to one or more other molecular entities,

- 34 040365 such as another antibody (for example, a bispecific antibody or diantibody), a detectable agent, a cytotoxic agent, a pharmaceutical agent, and/or a protein or peptide that can mediate the association of an antibody or antibody portion with another molecule (such as a streptavidin or polyhistidine label).

Some types of derivatized antibody molecule are obtained by cross-linking two or more antibodies (of the same type or different types, for example, to obtain bispecific antibodies). Suitable crosslinkers include crosslinkers that are heterobifunctional, having two separate reactive groups separated by an appropriate spacer (eg, mmalimidobenzoyl-M-hydroxysuccinimide ester) or homobifunctional (eg, disuccinimidyl suberate). Such linkers are available from Pierce Chemical Company, Rockford, 111.

Suitable detectable means by which an anti-TIM-3 antibody molecule can be derivatized (or labeled) include fluorescent compounds, various enzymes, prosthetic groups, luminescent materials, bioluminescent materials, fluorescent metal atoms, such as europium (Eu) and other lanthanides. , and radioactive materials (described below). Illustrative fluorescent detectable agents include fluorescein, fluorescein isothiocyanate, rhodamine, 5-dimethylamine-1-naphthalenesulfonyl chloride, phycoerythrin, and the like.

The antibody can also be derivatized with detectable enzymes such as alkaline phosphatase, horseradish peroxidase, β-galactosidase, acetylcholinesterase, glucose oxidase, and the like. When an antibody is derivatized with a detectable enzyme, it is detected by adding additional reagents that the enzyme uses to produce a detectable reaction product. For example, when the detectable agent horseradish peroxidase is present, the addition of hydrogen peroxide and diaminobenzidine results in a colored reaction product that is detectable. The antibody molecule can also be derivatized with a prosthetic group (eg streptavidin/biotin and avidin/biotin). For example, an antibody can be derivatized with biotin and can be detected by indirect measurement of avidin or streptavidin binding. Examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; and examples of bioluminescent materials include luciferase, luciferin, and aequorin.

The labeled antibody molecule can be used, for example, for diagnostic purposes and/or experimentally in a number of contexts, including (i) to isolate a given antigen by standard methods such as affinity chromatography or immunoprecipitation; (ii) to detect a given antigen (eg, in a cell lysate or cell supernatant) to assess the magnitude and profile of protein expression; (iii) to monitor tissue protein levels as part of a clinical testing procedure, for example to determine the effectiveness of a given treatment regimen.

An antibody molecule may be conjugated to another molecular entity, typically a label or therapeutic (eg, immunomodulatory, immunostimulatory, cytotoxic, or cytostatic) agent or moiety. In diagnostic or therapeutic applications, radioactive isotopes can be used. Radioactive isotopes that may be associated with anti-TIM-3 antibodies include, but are not limited to, α-, β- or γ-emitters or β- and γ-emitters. Such radioactive isotopes include, but are not limited to, 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 ( ^1u 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 Ga). Radioisotopes useful as drugs 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 Rh). Radioisotopes useful as labels, for example for diagnostic applications, 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.

The present invention relates to radiolabeled antibody molecules and methods for labeling them. In some embodiments, a method for labeling an antibody molecule is disclosed. The method includes bringing the antibody molecule into contact with a chelating agent, thereby obtaining a conjugated antibody. The conjugated antibody is radioactively labeled with a radioisotope such as ^1P indium, ⁹⁰ yttrium, and ¹⁷⁷ lutetium, thereby producing a labeled antibody molecule.

As discussed above, an antibody molecule may be conjugated to a drug. Therapeutically active radioisotopes have already been mentioned. Examples of other drugs include taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxy anthracine dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids , procaine, tatracaine,

- 35 040365 lidocaine, propranolol, puromycin, maytansinoids, such as maytansinol (see US patent No. 5208020), CC-1065 (see US patent No. 5475092, 5585499, 5846545) and their analogues or homologues. Drugs include, but are not limited to, antimetabolites (eg, methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine), alkylating agents (eg, mechlorethamine, thiotepa, chlorambucil, CC-1065, melphalan, carmustine (BSNU) and lomustine (CCNU), cyclophosphamide, busulfan, dibromomannite, streptozotocin, mitomycin C, and cis-dichlorodiaminplatinum(II) ((DDP) cisplatin), anthracyclines (eg, daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (eg, dactinomycin (formerly actinomycin), bleomycin, mithramycin and anthramycin (AMC)) and antimitotic agents (eg vincristine, vinblastine, taxol and maytansinoids).

In some aspects, the present invention relates to a method of providing a target-binding molecule that specifically binds to the TIM-3 receptor. For example, the target-binding molecule is an antibody molecule. The method includes: providing a target protein that contains at least a portion of a non-human protein, the portion being homologous (at least 70, 75, 80, 85, 87, 90, 92, 94, 95, 96, 97, 98 or 99% identical) to the corresponding human target protein, but differs by at least one amino acid (eg, at least one, two, three, four, five, six, seven, eight, or nine amino acids); obtaining an antibody molecule that specifically binds to the antigen; and evaluating the effectiveness of the binding compound in modulating the activity of the target protein. In addition, the method may include administering a binding compound (eg, an antibody molecule) or derivative (eg, a humanized antibody molecule) to a human.

In certain embodiments, the antibody molecule is a multispecific (eg, bispecific or trispecific) antibody molecule. Protocols for obtaining bispecific or heterodimeric antibody molecules are known in the art; including, but not limited to, for example, the ledge-in-cavity approach described, for example, in US 5,731,168; Fc pairing via electrostatic directional interaction as described, for example, in WO 09/089004, WO 06/106905 and WO 2010/129304; the formation of heterodimers of engineered chain exchange domains (SEEDs), as described, for example, in WO 07/110205; exchange of Fab arms as described, for example, in WO 08/119353, WO 2011/131746 and wO 2013/060867; a dual antibody conjugate, for example, by cross-linking to form a bispecific structure using a heterobifunctional reagent having an amine reactive group and a sulfhydryl reactive group, as described, for example, in US 4433059; bispecific antibody determinants obtained by recombination of half antibodies (heavy-light chain or Fab pairs) from different antibodies through a cycle of reduction and oxidation of disulfide bonds between the two heavy chains, as described, for example, in US 4444878; trifunctional antibodies, eg three Fab' fragments cross-linked via sulfhydryl reactive groups, as described, eg, in US5273743; biosynthetic binding proteins, for example a pair of scFvs, linked via C-terminal tails, preferably via disulfide or amine reactive chemical crosslinking, as described, for example, in US5534254; bifunctional antibodies, eg Fab fragments with different binding specificities, dimerized through leucine zippers (eg, c-fos and c-jun), which have a constant domain substituted, as described, for example, in US5582996; bispecific and oligospecific mono- and oligovalent receptors, e.g., the VH-CH1 regions of two antibodies (two Fab fragments) linked via a polypeptide spacer between the CH1 region of one antibody and the VH region of the other antibody, typically with their associated light chains as described , for example, in US5591828; bispecific DNA-antibody conjugates, eg cross-linking of antibodies or Fab fragments through a double-stranded DNA fragment, as described, for example, in US5635602; bispecific fusion proteins, for example an expression construct containing two scFvs with a hydrophilic helical peptide linker between them and a complete constant region, as described, for example, in US5637481; multivalent or multispecific binding proteins, e.g., a dimer of polypeptides having a first domain with an Ig heavy chain variable region binding region and a second domain with an Ig light chain variable region binding region, commonly referred to as diabodies (higher order structures are also contemplated, creating bispecific, trispecific, or tetraspecific molecules as described in, for example, US5837242, mini-antibody constructs with cross-linked VL and VH chains further linked via peptide spacers to the hinge and CH3 regions of the antibody, which can dimerize to form bispecific/multivalent molecules, as described in, for example, US5837821; VH and VL domains linked by a short peptide linker (e.g. 5 or 10 amino acids) or no linker at all in any orientation, which can form dimers to form bispecific diabodies; trimers and tetramers as described in, e.g., US5844094; VH strand -domains (or VL domains in family members) linked by peptide bonds with crosslinkable groups at the C-terminus, further linked to VL domains to form a series of FVs (or scFvs), as described, for example, in US5864019; and single-stranded binding polypeptides with both VH and VL domain linked via a peptide linker, which are combined into multivalent structures via non-covalent or chemical crosslinking

- 36 040365 formation of, for example, homobivalent, heterobivalent, trivalent and tetravalent structures using both scFV and diabody formats, as described, for example, in US5869620. Additional illustrative multispecific and bispecific molecules and methods for their preparation are presented in, for example, US5910573, US5932448, US5959083, US5989830, US6005079, US6239259, US6294353, US6333396, US6476198, US6511663, US66670458

US6833441, US7129330, US7183076, US7521056, US7527787, US7534866, US7612181, US2002004587A1,

US2002076406A1,

US2004220388A1,

US2005079170A1,

US2005266425A1,

US2007004909A1,

US2007274985A1,

US2008241884A1,

US2009155275A1,

US2002103345A1,

US2004242847A1,

US2005100543A1,

US2006083747A1,

US2007087381A1,

US2008050370A1,

US2008254512A1,

US2009162359A1,

US2003207346A1,

US2005003403A1,

US2005136049A1,

US2006120960A1,

US2007128150A1,

US2008069820A1,

US2008260738A1,

US2009162360A1,

US2003211078A1,

US2005004352A1,

US2005136051A1,

US2006204493A1,

US2007141049A1,

US2008152645A1,

US2009130106A1,

US2009175851A1,

US2004219643A1

US2005069552A1

US2005163782A1

US2006263367A1

US2007154901A1

US2008171855A1

US2009148905A1

US2009175867A1

US2009232811A1, US2009234105A1, US2009263392A1, US2009274649A1, EP346087A2, WO0006605A2, WO02072635A2, WO04081051A1, WO06020258A2, WO2985724

WO2007137760A2, WO2008119353A1, WO2009021754A2, WO2009068630A1, WO9103493A1,

WO9323537A1, WO9409131A1, WO9412625A2, WO9509917A1, WO9637621A2, WO9964460A1. The content of the above applications is incorporated into the present description by reference in its entirety.

In other embodiments, an anti-TIM-3 antibody molecule (e.g., a monospecific, bispecific, or multispecific antibody molecule) is covalently linked, e.g., fused, to another partner, e.g., a protein, e.g., one, two or more cytokines, e.g., as a fusion molecule, for example, a fusion protein. In other embodiments, the implementation of the fusion molecule contains one or more proteins, such as one, two or more cytokines. In one embodiment, the cytokine is an interleukin (IL) selected from one, two, three or more of IL-1, IL-2, IL-12, IL-15, or IL-21. In one embodiment, the bispecific antibody molecule has a first binding specificity for a first target (e.g., TIM-3), a second binding specificity for a second target (e.g., LAG-3 or PD-1), and optionally associated with an interleukin domain (e.g., IL -12), for example, full-length IL-12 or part of it. In other embodiments, the anti-TIM-3 antibody molecule is fused to another protein, eg one, two or more cytokines, eg as a fusion molecule. In other embodiments, the implementation of the fusion molecule contains one or more proteins, such as one, two or more cytokines. In one embodiment, the cytokine is an interleukin (IL) selected from one, two, three or more of IL-1, IL-2, IL-12, IL-15, or IL-21.

A fusion protein and a fusion polypeptide refer to a polypeptide having at least two parts covalently linked together, where each part is a polypeptide having a different property. The property may be a biological property such as in vitro or in vivo activity. The property can also be a simple chemical or physical property such as binding to a target molecule, catalyzing a reaction, etc. The two parts can be linked directly by one peptide bond or through a peptide linker, however, they are in the same reading frame with each other.

Exemplary Anti-TIM-3 Antibody Molecules

In certain embodiments, the anti-TIM-3 antibody comprises:

(a) a heavy chain variable region (VH) containing the amino acid sequence VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 10 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14;

(b) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 4 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7 and the amino acid sequence of VLCDR3 SEQ ID NO: 8;

(c) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 25 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14;

(d) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 24 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and VL containing the amino acid sequence of VLCDR1 SEQ ID NO:

- 37 040365

6, the amino acid sequence of VLCDR2 SEQ ID NO: 7 and the amino acid sequence of VLCDR3 SEQ ID NO: 8;

(e) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 31 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14; or (f) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 30 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7, and the amino acid sequence of VLCDR3 SEQ ID NO: 8.

In certain embodiments, the antibody molecule comprises a VH containing the VHCDR1 amino acid sequence selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 10 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14.

In certain embodiments, the antibody molecule comprises a VH containing the VHCDR1 amino acid sequence selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 4 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7, and the amino acid sequence of VLCDR3 SEQ ID NO: 8.

In certain embodiments, the antibody molecule comprises a VH containing the VHCDR1 amino acid sequence selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 25 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14.

In certain embodiments, the antibody molecule comprises a VH containing the VHCDR1 amino acid sequence selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 24 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7, and the amino acid sequence of VLCDR3 SEQ ID NO: 8.

In certain embodiments, the antibody molecule comprises a VH containing the VHCDR1 amino acid sequence selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 31; and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14.

In certain embodiments, the antibody molecule comprises a VH containing the VHCDR1 amino acid sequence selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 30 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7, and the amino acid sequence of VLCDR3 SEQ ID NO: 8.

In certain embodiments, an anti-TIM-3 antibody molecule comprises:

(i) a heavy chain variable region (VH) comprising the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3 or SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 4 or SEQ ID NO: 10 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and (ii) a light chain variable region (VL) comprising the VLCDR1 amino acid sequence of SEQ ID NO: 6 or SEQ ID NO: 12, the VLCDR2 amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 13, and the VLCDR3 amino acid sequence of SEQ ID NO: 8 or SEQ ID NO: 14.

In other embodiments, the anti-TIM-3 antibody molecule comprises:

(i) a heavy chain variable region (VH) comprising the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3 or SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 24 or SEQ ID NO: 25 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and (ii) a light chain variable region (VL) comprising the VLCDR1 amino acid sequence of SEQ ID NO: 6 or SEQ ID NO: 12, the VLCDR2 amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 13, and the VLCDR3 amino acid sequence of SEQ ID NO: 8 or SEQ ID NO: 14.

In other embodiments, the anti-TIM-3 antibody molecule comprises:

(i) a heavy chain variable region (VH) comprising the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3 or SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 30 or SEQ ID NO: 31 and the amino acid sequence of VHCDR3 SEQ ID

- 38 040365

NO: 5; and (ii) a light chain variable region (VL) comprising the VLCDR1 amino acid sequence of SEQ ID NO: 6 or SEQ ID NO: 12, the VLCDR2 amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 13, and the VLCDR3 amino acid sequence of SEQ ID NO: 8 or SEQ ID NO: 14.

In embodiments of the above antibody molecules, VHCDR1 comprises the amino acid sequence of SEQ ID NO: 3. In other embodiments, VHCDR1 comprises the amino acid sequence of SEQ ID NO: 9.

In embodiments of the above antibody molecules, VHCDR2 comprises the amino acid sequence of SEQ ID NO: 4. In other embodiments, VHCDR2 comprises the amino acid sequence of SEQ ID NO: 10. In other embodiments, VHCDR2 comprises the amino acid sequence of SEQ ID NO: 24. In other embodiments, VHCDR2 comprises amino acid sequence of SEQ ID NO: 25. In other embodiments, VHCDR2 comprises the amino acid sequence of SEQ ID NO: 30. In other embodiments, VHCDR2 comprises the amino acid sequence of SEQ ID NO: 31.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising an amino acid sequence at least 85% identical to any of SEQ ID NOs: 1, 16, 26, 32, 36, 44, 48, 52, 60, 68, 72, 76, 80, 84, 92 or 100.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 1, 16, 26, 32, 36, 44, 48, 52, 60, 68, 72, 76, 80, 84, 92, or 100.

In other embodiments, the aforementioned antibody molecules comprise a light chain variable domain containing an amino acid sequence at least 85% identical to any of SEQ ID NOs: 2, 20, 40, 56, 64, 88, 96, or 104.

In other embodiments, the aforementioned antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 2, 20, 40, 56, 64, 88, 96, or 104.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 1.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 16.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 18.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 26.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 28.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 32.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 34.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 36.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 38.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 44.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 46.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 48.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 50.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 52.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 54.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 60.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 62.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 68.

- 39 040365

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 70.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 72.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 74.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 76.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 78.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 80.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 82.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 84.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 86.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 92.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 94.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 100.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 102.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 116.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 121.

In other embodiments, the aforementioned antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 2.

In other embodiments, the aforementioned antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 20.

In other embodiments, the aforementioned antibody molecules comprise a light chain comprising the amino acid sequence of SEQ ID NO: 22.

In other embodiments, the aforementioned antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 40.

In other embodiments, the aforementioned antibody molecules comprise a light chain comprising the amino acid sequence of SEQ ID NO: 42.

In other embodiments, the aforementioned antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 56.

In other embodiments, the aforementioned antibody molecules comprise a light chain comprising the amino acid sequence of SEQ ID NO: 58.

In other embodiments, the aforementioned antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 64.

In other embodiments, the aforementioned antibody molecules comprise a light chain comprising the amino acid sequence of SEQ ID NO: 66.

In other embodiments, the aforementioned antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 88.

In other embodiments, the aforementioned antibody molecules comprise a light chain comprising the amino acid sequence of SEQ ID NO: 90.

In other embodiments, the aforementioned antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 96.

In other embodiments, the aforementioned antibody molecules comprise a light chain comprising the amino acid sequence of SEQ ID NO: 98.

In other embodiments, the aforementioned antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 104.

In other embodiments, the aforementioned antibody molecules comprise a light chain comprising the amino acid sequence of SEQ ID NO: 106.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 1 and the variable

- 40 040365 light chain domain containing the amino acid sequence of SEQ ID NO: 2.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 16 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 20.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 26 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 20.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 32 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 20.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 36 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 40.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 44 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 40.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 48 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 40.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 36 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 20.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 16 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 40.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 56.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 60 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 56.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 64.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 60 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 64.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 68 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 64.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 72 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 64.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 76 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 56.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 80 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 56.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 68 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 56.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 72 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 56.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 76 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 64.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 80 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 64.

In other embodiments, the aforementioned antibody molecules comprise a variable

- 41 040365 a heavy chain domain containing the amino acid sequence of SEQ ID NO: 84, and a light chain variable domain containing the amino acid sequence of SEQ ID NO: 88.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 92 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 96.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 100 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 104.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 18 and a light chain containing the amino acid sequence of SEQ ID NO: 22.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 28 and a light chain containing the amino acid sequence of SEQ ID NO: 22.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 34 and a light chain containing the amino acid sequence of SEQ ID NO: 22.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 38 and a light chain containing the amino acid sequence of SEQ ID NO: 42.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 46 and a light chain containing the amino acid sequence of SEQ ID NO: 42.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 50 and a light chain containing the amino acid sequence of SEQ ID NO: 42.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 116 and a light chain containing the amino acid sequence of SEQ ID NO: 22.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 121 and a light chain containing the amino acid sequence of SEQ ID NO: 42.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 54 and a light chain containing the amino acid sequence of SEQ ID NO: 58.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 62 and a light chain containing the amino acid sequence of SEQ ID NO: 58.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 54 and a light chain containing the amino acid sequence of SEQ ID NO: 66.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 62 and a light chain containing the amino acid sequence of SEQ ID NO: 66.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 70 and a light chain containing the amino acid sequence of SEQ ID NO: 66.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 74 and a light chain containing the amino acid sequence of SEQ ID NO: 66.

In other embodiments, the aforementioned antibody molecules comprise 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: 58.

In other embodiments, the aforementioned antibody molecules comprise 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: 58.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 70 and a light chain containing the amino acid sequence of SEQ ID NO: 58.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 74 and a light chain containing the amino acid sequence of SEQ ID NO: 58.

- 42 040365

In other embodiments, the aforementioned antibody molecules comprise 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: 66.

In other embodiments, the aforementioned antibody molecules comprise 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: 66.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 86 and a light chain containing the amino acid sequence of SEQ ID NO: 90.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 94 and a light chain containing the amino acid sequence of SEQ ID NO: 98.

In other embodiments, the aforementioned antibody molecules comprise a heavy chain containing the amino acid sequence of SEQ ID NO: 102 and a light chain containing the amino acid sequence of SEQ ID NO: 106.

In other embodiments, the aforementioned antibody molecules are selected from Fab, F(ab') ₂ , Fv, or a single chain Fv fragment (scFv).

In other embodiments, the aforementioned antibody molecules comprise a heavy chain constant region selected from IgG1, IgG2, IgG3, and IgG4.

In other embodiments, the aforementioned antibody molecules comprise a light chain constant region selected from the kappa and lambda light chain constant regions.

In some embodiments, the anti-TIM-3 antibody molecule comprises the CDR2 of the VH region of SEQ ID NO: 1 using the CDR definitions of Kabat or Chothia. In some embodiments, the anti-TIM-3 antibody molecule comprises a CDR2 and one or both of the CDR1 and CDR3 of the VH region of SEQ ID NO: 1 using the CDR definitions of Kabat or Chothia. In some embodiments, the anti-TIM-3 antibody molecule comprises the CDR2 of the VH region of SEQ ID NO: 1 in combination with the other 1, 2, 3, 4, or 5 (e.g., collectively, all) CDRs found in SEQ ID NO: 1 or SEQ ID NO: 2 using the CDR definitions according to Kabat or Chothia. In some embodiments, an anti-TIM-3 antibody molecule comprises VHCDR2 SEQ ID NO: 4. For example, an anti-TIM-3 antibody molecule may comprise VHCDR2 SEQ ID NO: 4 in combination with one or both of VHCDR1 SEQ ID NO: 3 and VHCDR3 SEQ ID NO: 5. As another example, an anti-TIM-3 antibody molecule may comprise the VHCDR2 of SEQ ID NO: 4 in combination with another 1, 2, 3, 4, or 5 (e.g., collectively, all) CDRs selected from SEQ ID NOs: 3, 5, 6, 7 and 8.

In some embodiments, the anti-TIM-3 antibody molecule comprises the CDR3 of the VL region of SEQ ID NO: 2 using the CDR definitions of Kabat or Chothia. In some embodiments, the anti-TIM-3 antibody molecule comprises a CDR3 and one or both of the CDR1 and CDR2 of the VL region of SEQ ID NO: 2 using the CDR definitions of Kabat or Chothia. In some embodiments, the anti-TIM-3 antibody molecule comprises the CDR3 of the VL region of SEQ ID NO: 2 in combination with the other 1, 2, 3, 4, or 5 (e.g., collectively, all) CDRs found in SEQ ID NO: 1 or SEQ ID NO: 2, using the CDR definitions according to Kabat of Chothia. In some embodiments, an anti-TIM-3 antibody molecule comprises VLCDR3 SEQ ID NO: 8. For example, an anti-TIM-3 antibody molecule may comprise VLCDR3 SEQ ID NO: 8 in combination with one or both of VHCDR1 SEQ ID NO: 6 and VHCDR2 SEQ ID NO: 7. As a further example, an anti-TIM-3 antibody molecule may comprise the VLCDR3 of SEQ ID NO: 8 in combination with another 1, 2, 3, 4, or 5 (e.g., collectively, all) CDRs selected from SEQ ID NO: 3-7.

In some embodiments, an anti-TIM-3 antibody molecule comprises a VH region CDR2 of SEQ ID NO: 1 and a VL region CDR3 of SEQ ID NO: 2, optionally in combination with an additional 1, 2, 3, or 4 (e.g., collectively, all) CDRs found in SEQ ID NO: 1 and SEQ ID NO: 2 using the CDR definitions according to Kabat or Chothia. In certain embodiments, the anti-TIM-3 antibody molecule comprises VHCDR2 SEQ ID NO: 4 and VLCDR3 SEQ ID NO: 8, optionally in combination with an additional 1, 2, 3, or 4 (e.g., collectively, all) CDRs selected from SEQ ID NO: 3, 5, 6 or 7.

In some embodiments, an anti-TIM-3 antibody molecule comprises a heavy chain constant region, a light chain constant region, the heavy chain variable region, and a light chain variable region as shown in Table 1. 1-4 (for example, SEQ ID NO: 1 and SEQ ID NO: 2). In certain embodiments, an anti-TIM-3 antibody molecule comprises a heavy chain constant region, a light chain constant region, and 1, 2, 3, 4, 5, or 6 (eg, all) of the CDRs shown in Table 1. 1-4.

In some embodiments, the anti-TIM-3 antibody molecule contains all or part of the heavy chain sequence of SEQ ID NO: 1. For example, in some embodiments, the anti-TIM-3 antibody molecule contains amino acids 1-98, 1-107, or 1-118 of SEQ ID NO: 1. In some embodiments, an anti-TIM-3 antibody molecule comprises amino acids 1-98 of SEQ ID NO: 1, an hCDR3 region (e.g., SEQ ID NO: 5 or a sequence substantially identical thereto) and a region

- 43 040365

VHFW4 (eg, human VHFW4 region, homologous region of human D or J sequences, amino acids 108-118 of SEQ ID NO: 1, or a sequence substantially identical thereto). In some embodiments, the VHFW4 region has no more than 1 or 2 non-identity positions relative to amino acids 108-118 of SEQ ID NO: 1. In some embodiments, the VHFW4 region has no more than 3, 4, 5, 6, 7, 8, 9, or 10 positions non-identities relative to amino acids 108-118 of SEQ ID NO: 1. In some embodiments, the hCDR3 region has no more than 1 or 2 positions of non-identity relative to SEQ ID NO: 5.

In other embodiments, the aforementioned antibody molecules are capable of binding to human TIM-3 with a dissociation constant (K _D ) of less than 0.5 nM.

In some embodiments, the anti-TIM-3 antibody molecule is capable of independently binding to human TIM-3 and cynomolgus TIM-3 with high affinity. In some embodiments, high affinity refers to a K _D of less than 5, 2, 1, 0.5, 0.4, 0.3, 0.2, or 0.1 nM, such as about 0.3 to 0.01 nM, eg from about 0.2 to 0.05 nM, eg as measured by the Biacore method.

In other embodiments, the aforementioned antibody molecules bind to cynomolgus TIM-3 with a K _D of less than 10, 5, 4, 3, 2, or 1 nM, for example, as measured using the Biacore method, FACS assay, or ELISA.

In other embodiments, the aforementioned antibody molecules bind to human TIM-3 with a K _D less than 5, 2, 1, 0.5, 0.4, 0.3, 0.2, or 0.1 nM, such as when measured using Biacore method, FACS analysis or ELISA.

In embodiments, the aforementioned antibody molecules do not bind to mouse TIM-3.

In some embodiments, the antibody molecule binds to TIM-3 of a mammal, such as a human. For example, an antibody molecule specifically binds to an epitope, such as a linear or conformational epitope (eg, an epitope as described herein) on TIM-3. In some embodiments, the epitope is at least a portion of a human or cynomolgus TIM-3 IgV domain. In some aspects, it is advantageous to identify an antibody that binds with high affinity to human and cynomolgus monkey homologues of the protein of interest. This desired cross-reactivity allows the same antibody (or two antibodies with the same CDR or variable regions) to be tested in an animal model and then administered to human patients as a therapeutic agent.

In certain embodiments, the above antibody molecules do not cross-react with mouse TIM-3. In certain embodiments, the above antibody molecules are less cross-reactive with rat TIM-3. For example, cross-reactivity can be measured by the Biacore method or by a binding assay using cells that express TIM-3 (eg, TIM-3 expressing human 300.19 cells). In other embodiments, the aforementioned antibody molecules bind an extracellular Ig-like domain (eg, IgV domain) of TIM-3.

In some embodiments, the aforementioned anti-TIM-3 antibody molecules bind to one or more of: two residues adjacent to the N-terminus of the A chain, BC loop, CC' loop, F chain, FG loop, and TIM- G chain. 3, or one or more (e.g., two, five, ten, fifteen, twenty, twenty-five, thirty, thirty-five, or all) residues of two or more of: two residues adjacent to the N-terminus of the A-strand, BC loop, CC' loops, F-chains, FG loops, or TIM-3 G-chains. The TIM-3 F chain contains residues G106 through I112; The TIM-3 G chain contains residues E121 through K130; the FG loop of TIM-3 contains residues between the F chain and the G chain, for example, including residues Q113 to D120; the BC loop of TIM-3 contains residues between the B chain and the C chain, for example, including residues P37 to P50; two residues adjacent to the N-terminus of the A chain include residues V24 and E25; the CC' loop contains residues between the C chain and the C' chain, for example, including residues G56 to N65. In other embodiments, the aforementioned anti-TIM-3 antibody molecules bind to one or more of the: A chain, EF loop, C chain, C'C loop, or C chain. The A chain contains residues Y26 through E29; the EF loop contains residues between the E chain and the F chain, for example including residues E98 to S105; The C chain contains residues V51 through K55; the C'C loop contains residues between the C' chain and the C' chain, for example, including residues D71 to D74; and the C chain contains residues V75 to W78. TIM-3 residue numbering is described, for example, in FIG. 18. In one embodiment, anti-TIM-3 antibody molecules bind to one or more (e.g., two, five, ten, fifteen, twenty, twenty-five, thirty, thirty-five, or all) residues in the F chain, G chain, and CC'-loop of TIM-3.

In some embodiments, the aforementioned anti-TIM-3 antibody molecules reduce or inhibit plasma membrane permeation or PtdSer-dependent TIM-3 permeation. In some embodiments, the aforementioned anti-TIM-3 antibody molecules reduce or inhibit binding to the PtdSer TIM-3 ligand. In some embodiments, the aforementioned anti-TIM-3 antibody molecules reduce or inhibit binding to the TIM-3 ligand of HMGB1. In some embodiments, the aforementioned antibody molecules

- 44 040365 tive TIM-3 reduce or inhibit binding to the TIM-3 ligand CEACAM-1. In some embodiments, the aforementioned anti-TIM-3 antibody molecules reduce or inhibit binding to the TIM-3 ligand semaphorin-4A. In some embodiments, the aforementioned anti-TIM-3 antibody molecules do not reduce or inhibit binding to the TIM-3 ligand galectin-9.

In some embodiments, the anti-TIM-3 antibody molecule interacts with, e.g., binds, the surface of TIM-3 (e.g., one, two, three, five, eight, ten, fifteen, or more contiguous or discontinuous (e.g., non-contiguous) amino acid residues selected from Val24, Glu25, Thr41, Gly56, Ala57, Cys58, Pro59, Val60, Phe61, Glu121, Lys122, Phe123, Asn124, Leu125, Lys126 and/or Leu127).

In some embodiments, an anti-TIM-3 antibody molecule interacts with, e.g., binds, the surface of TIM-3 (e.g., one, two, three, five, eight, ten, fifteen, twenty, twenty-one, twenty-five or more continuous or discontinuous ( e.g. non-contiguous) amino acid residues selected from Val24, Glu25, Tyr26, Phe39, Tyr40, Thr41, Gly56, Ala57, Cys58, Pro59, Val60, Phe61, Ser105, Gly106, Ile107, Asn119, Asp120, Glu121, Lys122, Phe123, Asn124 , Leu125, Lys126, Leu127 and/or Val128) for example, as shown in detail in table. 13.

In some embodiments, an anti-TIM-3 antibody molecule interacts with, e.g., binds, the surface of TIM-3 (e.g., one, two, three, five, eight, ten, fifteen, twenty, twenty-one, twenty-five or more continuous or discontinuous ( e.g. non-contiguous) amino acid residues selected from Glu23, Val24, Glu25, Tyr26, Thr41, Pro42, Ala43, Ala44, Pro45, Gly46, Asn47, Leu48, Val49, Pro50, Val51, Cys52, Trp53, Gly54, Lys55, Gly56, Ala57 , Cys58, Pro59, Val60, Phe61, Glu121, Lys122, Phe123, Asn124, Leu125, Lys126 and/or Leu127).

In some embodiments, an anti-TIM-3 antibody molecule interacts with, e.g., binds, the surface of TIM-3 (e.g., one, two, three, five, eight, ten, fifteen, twenty, twenty-one, twenty-five or more continuous or discontinuous ( e.g. non-contiguous) amino acid residues selected from Val24, Glu25, Tyr26, Phe39, Tyr40, Thr41, Pro42, Ala43, Ala44, Pro45, Gly46, Asn47, Leu48, Val49, Pro50, Val51, Cys52, Trp53, Gly54, Lys55, Gly56 , Ala57, Cys58, Pro59, Val60, Phe61, Ser105, Gly106, Ile107, Asn119, Asp120, Glu121, Lys122, Phe123, Asn124, Leu125, Lys126, Leu127 and/or Val128).

In other embodiments, the anti-TIM-3 antibody molecule competes with CEACAM-1 for binding to TIM-3. In one embodiment, the anti-TIM-3 antibody molecule interacts with, e.g., binds, one, two, or more (all) of C58, N119, and K122 TIM-3, e.g., displaces or competes with CEACAM-1 for binding to these residues. In one embodiment, the anti-TIM-3 antibody molecule reduces or blocks hydrogen bonding between TIM-3 K122 and CEACAM-1 N42. Regarding CEACAM-1, CEACAM-1 has been shown to be a ligand for TIM-3 and is required for its ability to mediate T cell inhibition, which may play an important role in autoimmunity regulation and antitumor immunity (Huang, et al. (2014) Nature doi:10.1038/nature13848).

Inhibition of the interaction between TIM-3 and CEACAM-1 can be used with other immunomodulators described herein (eg, an anti-PD-1 inhibitor) to enhance the immune response against cancer.

In another embodiment, the anti-TIM-3 antibody molecule interacts with, eg, binds to, a TIM-3 PtdSer-binding loop, eg, a human TIM-3 IgV domain. In one embodiment, the anti-TIM-3 antibody molecule interacts with, e.g., binds, at least two TIM-3 PtdSer-binding loops, e.g., the FG loop and the TIM-3 CC' loop (e.g., metal ion dependent ligand binding site (MILIBS )). For example, the carboxyl group of PtdSer can bind to the CC' loop of TIM-3 and the amino group of PtdSer can bind to the FG loop of TIM-3. In one embodiment, the anti-TIM-3 antibody molecule reduces or prevents PtdSer-mediated penetration of TIM-3 across the membrane. Thus, an anti-TIM-3 antibody molecule can reduce the recruitment of TIM-3 expressing cells and/or membrane penetration of apoptotic cells (which may expose PtdSer) for uptake.

In another embodiment, the anti-TIM-3 antibody molecule competes with HMGB1 for binding to TIM-3. For example, it reduces HMGB1 binding to TIM-3 residue 62 (Q in mouse TIM-3, E in human TIM-3). With respect to HMGB1, it has been reported to interact with TIM-3 to help tumor-associated dendritic cells suppress the nucleic acid-mediated innate immune response (Chiba et al., (2012) Nat. Immunol. 13(9): 832-842) . Thus, an anti-TIM-3 antibody molecule can enhance a nucleic acid-mediated innate immune response.

In another embodiment, the TIM-3 antibody molecule does not compete with or reduce the binding of the galectin-9 (Gal-9) ligand to TIM-3.

In embodiments, the anti-TIM-3 antibody molecule is a monospecific antibody molecule or a bispecific antibody molecule. In embodiments, the molecule

- 45 040365 la antibody has a first binding specificity for TIM-3 and a second binding specificity for PD-1, LAG-3, CEACAM (for example, CEACAM-1 and/or CEACAM-5), PD-L1 or PD-L2 . In an embodiment, the antibody molecule comprises an antigen-binding fragment of an antibody, such as a half-antibody or an anti-gnathic-binding fragment of a half-antibody.

In other embodiments, the aforementioned antibody molecules are capable of enhancing the response of antigen-specific T cells.

Within the scope of the present invention, an isolated nucleic acid molecule encoding the aforementioned antibody molecule, its vectors and host cells is provided. A nucleic acid molecule includes, but is not limited to, RNA, genomic DNA, and cDNA.

In embodiments, the isolated nucleic acid encodes a heavy chain variable region, or a light chain variable region, or both, of any of the aforementioned antibody molecules.

In other embodiments, the isolated nucleic acid 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: 11, 17, 29, 33, 37, 45, 49, 53, 61 , 69, 73, 77, 81, 85, 93, 101, 115 or 120.

In other embodiments, the isolated nucleic acid comprises a nucleotide sequence encoding a heavy chain variable domain, wherein the nucleotide sequence comprises any of SEQ ID NOs: 11, 17, 27, 33, 37, 45, 49, 53, 61, 69, 73, 77 , 81, 85, 93, 101, 115 or 120.

In other embodiments, the isolated nucleic acid comprises a nucleotide sequence encoding a heavy chain, wherein the nucleotide sequence is at least 85% identical to any of SEQ ID NOs: 19, 29, 35, 39, 47, 51, 55, 63, 71, 75 , 79, 83, 87, 95, 103, 117 or 122.

In other embodiments, the isolated nucleic acid comprises a nucleotide sequence encoding a heavy chain, wherein the nucleotide sequence comprises any of SEQ ID NOs: 19, 29, 35, 39, 47, 51, 55, 63, 71, 75, 79, 83, 87 , 95, 103, 117 or 122.

In other embodiments, the isolated nucleic acid 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: 15, 21, 41, 57, 65, 89, 97, 105, 118 , 123, 125 or 127.

In other embodiments, the isolated nucleic acid comprises a nucleotide sequence encoding a light chain variable domain, wherein the nucleotide sequence comprises any of SeQ ID NOs: 15, 21, 41, 57, 65, 89, 97, 105, 118, 123, 125, or 127 .

In other embodiments, the isolated nucleic acid comprises a nucleotide sequence encoding a light chain, wherein the nucleotide sequence is at least 85% identical to any of SEQ ID NOs: 23, 43, 59, 67, 91, 99, 107, 119, 124, 126 or 128.

In other embodiments, the isolated nucleic acid comprises a nucleotide sequence encoding a light chain, wherein the nucleotide sequence comprises any of SEQ ID NOs: 23, 43, 59, 67, 91, 99, 107, 119, 124, 126, or 128.

Pharmaceutical compositions and kits

In some aspects, the present invention relates to compositions, such as pharmaceutically acceptable compositions, which comprise the anti-TIM-3 antibody molecule described herein, together with a pharmaceutically acceptable carrier. As used herein, a pharmaceutically acceptable carrier includes any and all diluents, dispersion media, isotonicity and absorption delaying agents, and the like, which are physiologically compatible. The carrier may be suitable for intravenous, intramuscular, subcutaneous, parenteral, intraspinal, or epidermal administration (eg, by injection or infusion).

The compositions described in the present description may take various forms. They include, for example, liquid, semi-solid and solid dosage forms such as liquid solutions (eg injectable and infusible solutions), dispersions or suspensions, liposomes and suppositories. The appropriate form depends on the intended route of administration and therapeutic application. Typical suitable compositions are in the form of injectable or infusible solutions. One suitable route of administration is parenteral (eg, intravenous, subcutaneous, intraperitoneal, intramuscular). In some embodiments, the antibody molecule is administered by intravenous infusion or injection. In certain embodiments, the antibody is administered via intramuscular or subcutaneous injection.

The terms parenteral administration and parenterally administered, as used herein, mean routes of administration other than enteral and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal,

- 46 040365 intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intravertebral, epidural and intrasternal injection and infusion.

Therapeutic compositions generally must be sterile and stable under the conditions of manufacture and storage. The composition may be formulated as a solution, microemulsion, dispersion, liposome, or other ordered structure suitable for high antibody concentrations. Sterile injectable solutions can be prepared by incorporating the active compound (ie, antibody or antibody portion) in the required amount in an appropriate solvent with one or a combination of the ingredients listed above, if necessary, followed by filtered sterilization. Typically, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains the basic dispersion medium and the other required ingredients from the ingredients listed above. In the case of sterile powders for the preparation of sterile injectable solutions, vacuum drying and freeze drying are preferred methods of manufacture, which provide a powder of the active ingredient with any additional required ingredient from its previously filter-sterilized solution. Proper fluidity of the solution can be maintained, for example, using coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by using surfactants. Prolonged absorption of an injectable composition can be achieved by including in the composition an agent that delays absorption, such as monostearates and gelatin.

Antibody molecules can be administered in a variety of ways. Several such routes are known in the art, and for many therapeutic applications, intravenous injection or infusion is a suitable route/mode of administration. In one embodiment, the antibody molecules can be administered via intravenous infusion at a rate greater than 20 mg/min, such as 20-40 mg/min, and preferably greater than or equal to 40 mg/min to achieve a dose of about 35 to 440 mg/m ² , preferably from about 70 to 310 mg/m ² and more preferably from about 110 to 130 mg/m ² . In one embodiment, the antibody molecules can be administered by intravenous infusion at a rate of less than 10 mg/min; preferably less than or equal to 5 mg/min to achieve a dose of about 1 to 100 mg/m ² , preferably about 5 to 50 mg/m ² , about 7 to 25 mg/m ² and more preferably about 10 mg/m ² . As will be appreciated by one of skill in the art, the route and/or route of administration will vary depending on the desired results. In certain embodiments, the active compound can be formulated with a carrier that protects the compound from rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters and polylactic acid can be used. Many methods for preparing such formulations are patented or well 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.

In certain embodiments, the antibody molecule may be administered orally, for example, with an inert diluent or digestible food carrier. The antibody molecule (and other ingredients, if desired) may also be enclosed in a hard or soft gelatin capsule, compressed into tablets, or incorporated directly into the individual's diet. For oral therapeutic administration, the antibody molecule may be incorporated with excipients and used in the form of swallowable tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, wafers, and the like. For administration of the antibody molecule by a route other than parenteral administration, it may be necessary to coat the compound or co-administer the compound with the material to prevent its inactivation. Therapeutic compositions can also be administered using medical devices, and several such devices are known in the art.

Dosing regimens are adjusted to provide the desired response (eg, therapeutic response). For example, a single bolus may be administered, several divided doses may be administered over time, or the dose may be proportionally reduced or increased as necessary for the therapeutic case. It is particularly advantageous to formulate parenteral compositions in unit dosage form for ease of administration and dosage uniformity. Unit dosage form, as used in the present invention, refers to physically discrete units suitable as single dosages for individuals to be treated; wherein each unit contains a predetermined amount of the active compound, calculated to achieve the desired therapeutic effect, together with the required pharmaceutical carrier. The characteristics of the unit dosage forms of the present invention are determined by and are directly dependent on (a) the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in this area in relation to the formulation of the active compound for the treatment of sensitivity of individuals.

An exemplary non-limiting range for a therapeutically or prophylactically effective amount of an antibody molecule is 0.1-30 mg/kg, more preferably 1-25 mg/kg. Dozi

The patterns and therapeutic regimens of an anti-TIM-3 antibody molecule can be determined by the skilled artisan. In certain embodiments, the anti-TIM-3 antibody molecule is administered by injection (e.g., subcutaneously or intravenously) at a dose of about 1 to 40 mg/kg, e.g., 1 to 30 mg/kg, e.g., about 5 to 25 mg/kg, about 10 to 20 mg/kg, about 1 to 5 mg/kg, 1 to 10 mg/kg, 5 to 15 mg/kg, 10 to 20 mg/kg, 15 to 25 mg/kg, or approximately 3 mg/kg. The dosing schedule may vary, for example, from once a week to once every 2, 3 or 4 weeks. In one embodiment, the anti-TIM-3 antibody molecule is administered at a dose of about 10 to 20 mg/kg biweekly. The antibody molecule can be administered by intravenous infusion at a rate greater than 20 mg/min, such as 20-40 mg/min and preferably greater than or equal to 40 mg/min to achieve a dose of about 35 to 440 mg/m ² , preferably about 70 to 310 mg/m ² and more preferably from about 110 to 130 mg/m ² . In embodiments, an infusion rate of about 110 to 130 mg/m ² provides a level of about 3 mg/kg. In other embodiments, the antibody molecule can be administered via intravenous infusion at a rate of less than 10 mg/min, such as less than or equal to 5 mg/min, to achieve a dose of about 1 to 100 mg/m ² , such as about 5 to 50 mg/m ² , from about 7 to 25 mg/m ² or about 10 mg/m ² . In some embodiments, the antibody is infused over a period of approximately 30 minutes. It should be noted that dosage values may vary depending on the type and severity of the condition to be alleviated. In addition, it should be understood that for any particular individual, specific dosage regimens should be adjusted over time in accordance with the needs of the individual and the professional opinion of the person who administers or supervises the administration of the compositions, and that the dosage ranges described herein are only illustrative and are not intended to limit the scope or practice of the claimed composition.

Pharmaceutical compositions within the scope of the present invention comprise a therapeutically effective amount or a prophylactically effective amount of an antibody molecule. A therapeutically effective amount refers to an amount effective, at the required dosages and for the required periods of time, to achieve the desired therapeutic result. The therapeutically effective amount of the modified antibody or antibody fragment may vary depending on such factors as the disease, age, sex and body weight of the individual and the ability of the antibody or antibody portion to induce the desired response in the individual. A therapeutically effective amount is also one in which any toxic or deleterious effects of the antibody molecule are outweighed by the therapeutically beneficial effects. A therapeutically effective dosage preferably inhibits the measurable parameter by at least about 20%, more preferably at least about 40%, even more preferably at least about 60%, and even more preferably at least about 80% relative to untreated treatment of individuals. The measurable parameter can be, for example, the growth rate of a tumor or the growth rate of a pathogen. The ability of a compound to inhibit a measurable parameter can be assessed in an animal model system predictive of efficacy in the corresponding disease in humans. Alternatively, this property of the composition can be assessed by testing the ability of the compound to inhibit, such as in vitro inhibition, using assays known to the skilled artisan.

A prophylactic effective amount refers to an amount effective, at the required dosages and for the required periods of time, to achieve the desired prophylactic result. Generally, since a prophylactic dose is used in individuals before or at an early stage of the disease, the prophylactically effective amount is less than the therapeutically effective amount.

Also within the scope of the present invention is a kit containing an antibody molecule as described herein. The kit may include one or more elements, including: instructions for use; other reagents, such as a label, drug, or agent suitable for chelating or otherwise binding the antibody to the label, or drug, or radioprotective composition; devices or other materials for preparing an antibody molecule for administration; pharmaceutically acceptable carriers; and devices or other materials for administration to an individual.

Application of Anti-TIM-3 Antibody Molecules

TIM-3 is a co-inhibitory protein expressed, for example, on activated T-helper 1 (Th1) CD4+ and cytotoxic CD8+ T cells that secrete IFN-γ. TIM-3 is largely co-expressed on PD-1+ depleted T cells, as shown in preclinical models of cancer and viral depletion. Co-blockade of these cascades may restore effector T cell function (eg, IFN-γ secretion, proliferation) in several models, as well as in human PBMCs derived from patients with metastatic melanoma and patients

- 48 040365 with HIV or HCV. TIM-3 is also abundant on native FoxP3+ regulatory T cells (and FoxP3-negative induced regulatory cells), and nTreg expression correlates with disease severity in NSCLC, hepatic cell carcinoma, and ovarian carcinoma. In TIM-3+ mouse models, nTregs have been shown to be more immunosuppressive (secreting higher levels of IL-10 and TGF-β).

In addition, TIM-3 may play an important role on innate immune cells, including NK cells, monocytes/macrophages, and dendritic cells (DCs). TIM-3 is constitutively expressed on macrophages and DCs, and its blockade can enhance TNF-α secretion from human monocytes and increase NF-κB expression in a mouse dendritic cell line. TIM-3 may also contribute to the expansion of myeloid-derived suppressor cells (MDSCs). Constitutive expression of TIM-3 on macrophages is associated with less IL-12 secretion, and suppression of TIM-3 activation after TLR may lead to increased IL-12 secretion and subsequent effector T cell responses.

The antibody molecules described herein have in vitro and in vivo diagnostic utility as well as therapeutic and prophylactic utility. In some embodiments, the antibody molecules modulate (eg, enhance or inhibit) the immune response in an individual through TIM-3 binding. For example, these molecules can be administered to cells in culture, in vitro or ex vivo, or to an individual, eg, a human, eg, in vivo, to modulate (eg, enhance or inhibit) immunity.

Thus, in some aspects, the invention relates to a method for modifying an immune response in an individual, comprising administering to the individual an antibody molecule described herein such that modification of the immune response in the individual occurs. In some embodiments, the implementation of the immune response is enhanced, stimulated or activated. In certain embodiments, the implementation of the immune response is inhibited or suppressed. For example, these antibody molecules can be administered to cells in culture, eg in vitro or ex vivo, or to the individual, eg in vivo, for the treatment, prevention and/or diagnosis of various disorders such as cancers, immune disorders and infectious diseases.

As used herein, the term individual includes human and non-human animals. In some embodiments, the subject is a human, such as a human patient, having a disorder or condition characterized by abnormal TIM-3 functioning. Typically, an individual has at least a portion of TIM-3, including a TIM-3 epitope, that is bound by the antibody molecule, eg, a sufficiently high level of protein and epitope to support binding of the antibody to TIM-3. The term non-human animals includes mammals and non-mammals such as non-human primates. In some embodiments, the individual is a human. In some embodiments, the subject is a human patient in need of enhanced immune response. The methods and compositions described herein are useful in the treatment of human patients having a disorder that can be treated by modulating (eg, enhancing or inhibiting) an immune response.

Methods for the treatment of immune disorders.

TIM-3 is a transmembrane receptor expressed on T cells, such as CD4+ T cells, CD8+ T cells, regulatory T cells, and differentiated Th1 cells. TIM-3 dependent transport of Th1 cells into the target tissue can be inhibited by soluble TIM-3 (see US 7470428). Thus, modulating TIM-3 function can reduce T cell transport to the target tissue, for example, in individuals with an autoimmune disease. TIM-3 may play an important role in the induction of autoimmune diseases through the regulation of macrophage activation and/or function. Thus, in certain embodiments, the anti-TIM-3 antibody molecules described herein are useful for use in suppressing an unwanted immune response, such as in the treatment of autoimmune diseases.

Moreover, as described in the Examples section of this specification, anti-TIM-3 antibodies can stimulate NK cell-mediated killing of target cells and can enhance IFN-gamma secretion and CD4+ T cell proliferation. Thus, in certain embodiments, the anti-TIM-3 antibody molecules described herein are useful for stimulating a desired immune response, such as an immune response against a cancer cell or pathogen.

Anti-TIM-3 antibodies described herein can be used to treat immune disorders, especially T-lymphocyte-mediated disorders, including, but not limited to, chronic inflammatory diseases and disorders such as Crohn's disease, reactive arthritis, including Lyme disease, insulin-dependent diabetes, organ-specific autoimmunity, including multiple sclerosis, Hashimoto's thyroiditis and Graves' disease, contact dermatitis, psoriasis, graft rejection, graft-versus-host disease, sarcoidosis, atopic conditions such as asthma and allergies, including allergic rhinitis, gastrointestinal intestinal allergies, including food

- 49 040365 allergies, eosinophilia, conjunctivitis, glomerular nephritis (eg IgA nephropathy), sensitivity to certain pathogens such as helminths (eg leishmaniasis).

In certain embodiments, an anti-TIM-3 antibody is used to modulate T cell function, eg, CD4+ T cell, CD8+ T cell, Treg, Th17, and Th1 function. In some embodiments, an anti-TIM-3 antibody molecule causes blockade of TIM-3 and is used to treat an immune disorder that is not a Th1 dependent disease (see Schroll et al., Am J Pathol 2010 April; 176(4):1716 -1742). In certain embodiments, the anti-TIM-3 antibody molecule does not cause blockade of TIM-3.

In some aspects, the present invention provides methods for administering an anti-TIM-3 antibody molecule resulting in stimulation or reduction of T cell transport into a target tissue, stimulation or inhibition of antigen presenting cell (APC) activation.

In some embodiments, the individual is in need of treatment for an autoimmune disease. Autoimmune disease includes diseases in which the individual's own antibodies react with host tissue or in which immune effector T cells are autoreactive against endogenous self peptides and cause tissue destruction. Thus, an immune response is induced against the individual's own antigens, called self-antigens. Autoimmune diseases include, but are not limited to, rheumatoid arthritis, Crohn's disease, e.g., childhood Crohn's disease, multiple sclerosis, systemic lupus erythematosus (SLE), autoimmune encephalomyelitis, myasthenia gravis (MG), Hashimoto's thyroiditis, Goodpasture's syndrome, pemphigus (e.g., pemphigus vulgaris), Graves' disease, autoimmune hemolytic anemia, autoimmune thrombocytopenic purpura, scleroderma with anti-collagen antibodies, mixed connective tissue disease, polymyositis, pernicious anemia, idiopathic Addison's disease, autoimmunity-associated infertility, glomerulonephritis (eg, sickle-shaped glomerulonephritis, proliferative glomerulonephritis), proliferative glomerulonephritis bullous pemphigoid, Sjögren's syndrome, insulin resistance, autoimmune diabetes mellitus (type 1 diabetes mellitus; insulin-dependent diabetes mellitus), atherosclerosis, and Alzheimer's disease.

In some aspects, the anti-TIM-3 antibody molecule described herein is administered to treat an unwanted immune response to an allergen. Examples of natural animal and plant allergens include proteins specific to the following genera: Canine (Canis familiaris); Dermatophagoides (eg Dermatophagoides farinae); Felis (Felis domesticus); Ambrosia (Ambrosia artemiisfolia; Lolium (e.g. Lolium perenne or Lolium multiflorum); Cryptomeria (Cryptomeria japonica); Alternaria (Alternaria alternata); Alder; Alnus (Alnus gultinosa); Betula (Betula verrucosa); Quercus (Quercus alba); Olea ( Olea europa); Artemisia (Artemisia vulgaris); Plantago (e.g. Plantago lanceolata); Parietaria (e.g. Parietaria officinalis or Parietaria judaica); Blattella (e.g. Blattella germanica); Apis (e.g. Apis multiflorum); Cupressus (e.g. Cupressus sempervirens, Cupressus arizonica and Cupressus macrocarpa); Juniperus (eg Juniperus sabinoides, Juniperus virginiana, Juniperus communis and Juniperus ashei); Thuya (eg Thuya orientalis); Chamaecyparis (eg Chamaecyparis obtusa); Periplaneta (eg Periplaneta americana); Agropyron (e.g. Agropyron repens); Secale (e.g. Secale cereale); Triticum (e.g. Triticum aestivum); Dactylis (e.g. Dactylis glomerata); Festuca (e.g. Festuca elat ior); Poa (for example, Poa pratensis or Poa compressa); Avena (for example, Avena sativa); Holcus (for example, Holcus lanatus); Anthoxanthum (eg Anthoxanthum odoratum); Arrhenatherum (for example, Arrhenatherum elatius); Agrostis (eg Agrostis alba); Phleum (for example, Phleum pratense); Phalaris (eg Phalaris arundinacea); Paspalum (for example, Paspalum notatum); Sorghum (eg Sorghum halepensis) and Bromus (eg Bromus inermis).

In some embodiments, an anti-TIM-3 antibody molecule is administered to treat multiple sclerosis, Crohn's disease, sepsis, SIRS (systemic inflammatory response syndrome), or glomerulonephritis.

Methods for the treatment of malignant tumors.

In some aspects, the present invention provides methods for administering an anti-TIM-3 antibody molecule for the treatment of cancer. Without wishing to be bound by theory, in some embodiments, the anti-TIM-3 antibody molecule stimulates the patient's immune system to recognize and destroy cancer cells, thereby treating the cancer. In some embodiments, the cancer being treated expresses TIM-3 and the anti-TIM-3 antibody molecule targets the cancer cells or cells in the tumor microenvironment.

In some aspects, the present invention relates to in vivo treatment of an individual with an anti-TIM-3 antibody molecule such that the growth of malignant tumors is inhibited. Anti-TIM-3 antibody can be used alone to inhibit the growth of malignant tumors. Alternatively, an anti-TIM-3 antibody may be used in combination with one or more of: a standard cancer treatment (eg, for malignant or infectious disorders) or another antibody or antigen-binding fragment thereof, an immunomodulator (eg, a co-stimulatory molecule activator or an inhibitor of an inhibitory

- 50 040365 molecules); vaccines (eg, cancer vaccine); or other forms of cellular immunotherapy as described below.

Thus, in some embodiments, the invention relates to a method of inhibiting tumor cell growth in an individual, comprising administering to the individual a therapeutically effective amount of an anti-TIM-3 antibody molecule described herein.

In some embodiments, the methods are useful for treating cancer in vivo. To achieve antigen-specific enhancement of immunity, the anti-TIM-3 antibody molecule can be administered together with the antigen of interest. When anti-TIM-3 antibodies are administered in combination with one or more agents, the combination may be administered in any order or simultaneously.

Types of cancer

In some aspects, a method is provided for treating, e.g., reducing or alleviating, a hyperproliferative condition or disorder (e.g., cancer), e.g., a solid tumor, hematologic cancer, soft tissue tumor, or metastatic lesion, in an individual. The method includes administering to an individual one or more of the anti-TIM-3 antibody molecules described herein, alone or in combination with other agents or therapeutic moieties.

As used herein, the term cancer is intended to include all types of malignant growth or oncogenic processes, metastatic tissues or malignantly transformed cells, tissues or organs, regardless of histopathological type or degree of invasiveness. Examples of malignant disorders include, but are not limited to, solid tumors, hematologic malignancies, soft tissue tumors, and metastatic lesions. Examples of solid tumors include malignant tumors such as sarcomas and carcinomas (including adenocarcinomas and squamous cell carcinomas) of various organ systems such as tumors affecting the liver, lung, breast, lymphoid organs, gastrointestinal tract (eg colon), urinary tract (eg, kidneys, urothelial cells), prostate, and pharynx. Adenocarcinomas include malignancies such as most types of colon cancer, rectal cancer, renal cell carcinoma, liver cancer, non-small cell lung carcinoma, small intestine cancer, and esophageal cancer. Squamous cell carcinoma includes malignant tumors, for example, in the lung, esophagus, skin, head and neck, mouth, anus, and cervix. In one embodiment, the cancer is a melanoma, such as advanced stage melanoma. Metastatic lesions of the aforementioned cancers can also be treated or prevented using the methods and compositions described herein.

Exemplary cancers whose growth is inhibited using the antibody molecules described herein include cancers that typically respond to immunotherapy. Non-limiting examples of cancers suitable for treatment include melanoma (eg, metastatic malignant melanoma), kidney cancer (eg, clear cell carcinoma), prostate cancer (eg, hormone-refractory adenocarcinoma of the prostate), breast cancer, colon cancer, and lung cancer. (eg, non-small cell lung cancer). In addition, refractory or recurrent cancers can be treated using the antibody molecules described herein.

Malignant tumors include, but are not limited to, basal cell carcinoma, biliary tract cancer; bladder cancer; bone cancer; cancer of the brain and central nervous system; primary CNS lymphoma; neoplasm of the central nervous system (CNS); mammary cancer; cervical cancer; choriocarcinoma; cancer of the colon and rectum; malignant tumor of the connective tissue; malignant tumor of the digestive system; endometrial cancer; esophageal carcinoma; malignant tumor of the eye; head and neck cancer; stomach cancer; intraepithelial neoplasm; kidney cancer; throat cancer; leukemia (including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic or acute leukemias); liver cancer; lung cancer (eg, small cell and non-small cell); lymphoma, including Hodgkin's lymphoma and non-Hodgkin's lymphoma; lymphocytic lymphoma; melanoma, such as cutaneous or intraocular malignant melanoma; myeloma; neuroblastoma; a malignant tumor of the oral cavity (eg, lips, tongue, oral cavity and pharynx); ovarian cancer; pancreas cancer; prostate cancer; retinoblastoma; rhabdomyosarcoma; rectal cancer; malignant tumor of the respiratory system; sarcoma; skin cancer; stomach cancer; testicular cancer; thyroid cancer; cancer of the body of the uterus; malignant tumor of the urinary system, hepatocarcinoma, cancer of the anal area, carcinoma of the fallopian tubes, carcinoma of the vagina, carcinoma of the external female genital organs, cancer of the small intestine, cancer of the endocrine system, cancer of the parathyroid gland, cancer of the adrenal gland, soft tissue sarcoma, cancer of the urethra, cancer of the penis, childhood solid tumors, spinal tumor, brainstem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, environmentally induced malignancies, including

- 51 040365 tea malignant tumors induced by asbestos, as well as other carcinomas and sarcomas and combinations of these malignant tumors.

In some embodiments, the cancer to be treated with antibody molecules includes, but is not limited to, solid tumors, hematologic malignancies, soft tissue tumors, and metastatic lesions. Examples of solid tumors include malignant tumors such as sarcomas, adenocarcinomas, and carcinomas of various organ systems such as sarcomas, adenocarcinomas, and carcinomas affecting the lung, breast, lymphoid organs, gastrointestinal tract (eg, large intestine), genitals, and genitourinary tract (eg, kidney cells, urothelium, bladder cells), pharynx, CNS (eg, brain, neuronal or glial cells), skin (eg, melanoma), and pancreas, as well as adenocarcinomas, which include malignancies such as most types of colon cancer, rectal cancer, renal cell carcinoma, liver cancer, non-small cell lung carcinoma, small intestine cancer, and esophageal cancer. The methods and compositions described herein are also useful in the treatment of metastatic lesions associated with the aforementioned cancers.

Without wishing to be bound by theory, in some embodiments, a patient is more likely to respond to treatment with an immunomodulator (optionally in combination with one or more agents as described herein) if the patient has a cancer that highly expresses PD-L1, and /or if the cancer is infiltrated with antitumor immune cells, eg TIL. Antitumor immune cells may be positive for CD8, PD-L1 and/or IFN-γ; thus, CD8, PD-L1 and/or IFN-γ levels can serve as an indicator of TIL levels in the microenvironment. In certain embodiments, the cancer microenvironment is referred to as triple positive for PD-L1/CD8/IFN-Y.

Thus, in some aspects, the present application relates to methods 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, for example, two or all three of the markers, then administering to the patient a therapeutically effective amount of an anti-PD-1 antibody molecule, optionally in combination with one or more other immunomodulators or anti-cancer agents, such as an anti-TIM3 antibody, as described herein.

In the following indications, a high proportion of patients are triple positive for PDL1/CD8/IFN-Y: lung cancer (squamous cell); lung cancer (adenocarcinoma); head and neck cancer; stomach cancer; NSCLC; HNSCC; gastric cancer (eg, MSIhi and/or EBV+); CRC (for example, MSIhi); nasopharyngeal cancer (NPC); cervical cancer (eg, squamous cell); thyroid cancer, such as papillary thyroid cancer; melanoma; breast cancer TN; and DLBCL (diffuse large B-cell lymphoma). In breast cancer in general and colon cancer in general, a moderate proportion of patients are triple positive for PD-L1/CD8/IFN-γ. In the following indications, a small proportion of patients are triple positive for PDL1/CD8/IFN-Y: ER+ breast cancer and pancreatic cancer. These data are discussed further in Example 9. Regardless of whether a large or small proportion of patients are triple positive for these markers, patient screening for these markers identifies the proportion of patients who have a particularly high likelihood of a favorable response to anti-PD-1 antibody therapy ( for example, an anti-PD-1 blocking antibody), optionally in combination with one or more other immunomodulators (for example, an anti-TIM-3 antibody molecule described herein, an anti-LAG-3 antibody molecule, or an anti-PD-L1 antibody molecule) and/ or means against a malignant tumor, for example, the means shown in table. 6 and described in the publications given in table. 6.

In some embodiments, the sample is classified as triple positive for PDL1/CD8/IFN-γ. This indicator can be roughly reduced to two thresholds: whether an individual cell is classified as positive and whether the sample as a whole is classified as positive. First, the level of PD-L1, CD8 and/or IFN-γ can be measured in an individual cell. In some embodiments, a cell that is positive for one or more of these markers is a cell that has a higher level of the marker than a control cell or reference value. For example, in some embodiments, a high level of PD-L1 in a given cell is a level greater than the level of PD-L1 in a corresponding non-cancerous tissue in a patient. As another example, in some embodiments, the high level of CD8 or IFN-γ in a given cell is the level of that protein normally seen in TIL. Second, it is also possible to measure the percentage of cells in a sample that are positive for PD-L1, CD8, and/or IFN-γ (it is not necessary that one cell expresses all three markers). In some embodiments, a triple positive sample is a sample that has a high percentage of cells, such as higher

- 52 040365 reference value or higher than in the control sample, which are positive for these markers.

In other embodiments, the levels of PDL1, CD8 and/or IFN-γ as a whole in the sample can be measured. In this case, a high level of CD8 or IFN-γ in the sample may represent the level of this protein commonly seen in a TIL-infiltrated tumor. Likewise, a high level of PD-L1 may represent the level of this protein commonly seen in a tumor sample, eg, in the tumor microenvironment.

The identification of patient subgroups that are PDL1/CD8/IFN-Y triple positive, as shown in Example 10 of the present disclosure, has identified certain patient subpopulations that are likely to respond particularly well to anti-PD-1 antibody therapy. For example, many IM-TN (immunomodulatory, triple negative) breast cancer patients are triple positive for PDL1/CD8/IFN-Y. 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 is a breast cancer that does not express the estrogen receptor (ER), progesterone receptor (PR), and Her2/neu. These cancers are difficult to treat because they usually do not respond to agents that target ER, PR, and Her2/neu. Triple negative breast cancer can be further subdivided into different classes, one of which is immunomodulatory. As described in Lehmann et al., IM-TN breast cancer is enriched in factors involved in immune cell processes, e.g., one or more of immune cell signaling (e.g., TH1/TH2 cascade, NK cell cascade, B signaling cascade α-cell receptor, DC cascade and T cell receptor signaling), cytokine signaling (eg, cytokine cascade, IL-12 cascade, and IL-7 cascade), antigen processing and presentation, signaling through central immune signaling cascades (eg , NFKB, TNF and JAK/STAT signaling), genes involved in T-cell function, immune transcription, interferon response (IFN) and antigen processing. Thus, in some embodiments, the cancer being treated is a cancer that is or has been determined to be positive for one or more IM-TN breast cancer markers, e.g., a factor that stimulates one or more from immune cell signaling (eg, TH1/TH2 cascade, NK cell cascade, B cell receptor signaling cascade, DC cascade, and T cell receptor signaling), cytokine signaling (eg, cytokine cascade, IL-12 cascade, and IL-7), antigen processing and presentation, signaling through central immune signaling cascades (eg, signaling through NFKB, TNF, and JAK/STAT), genes involved in T cell function, immune transcription, interferon (IFN) response, and processing antigen.

As another example, within the scope of the present invention, it has been shown that a subset of colon cancer patients with high MSI (microsatellite instability) is also triple positive for PD-L1/CD8/IFN-Y. Thus, in some embodiments, an anti-PD-1 antibody, optionally in combination with one or more immunomodulators, such as an anti-TIM-3 antibody described herein, an anti-LAG-3 antibody, or an anti-PD-L1 antibody, and one or more several anti-cancer agents, such as the anti-cancer agent described in Table 1. 6 or in the publication in table. 6 is administered to a patient who has or is identified as having high MSI colon cancer, thereby treating the cancer. In some embodiments, a high MSI cell is a cell having an MSI level above a reference value, or a value in a control cell, such as a non-cancerous cell of the same tissue type as the cancer.

As another example, the present invention shows that a subset of gastric cancer patients who have high MSI and/or who are EBV+ are also triple positive for PD-L1/CD8/IFN-Y. Thus, in some embodiments, an anti-PD-1 antibody, optionally in combination with one or more immunomodulators, such as an anti-TIM-3 antibody described herein, an anti-LAG-3 antibody, or an anti-PD-L1 antibody, and one or more anti-cancer agents, such as the anti-cancer agent described in Table 1. 6 or in the publication in table. 6 is administered to a patient who has or is identified as having gastric cancer with high MSI and/or EBV+, thereby treating the cancer. In some embodiments, a high MSI cell is a cell having an MSI level above a reference value or a value in a control cell, such as a non-cancerous cell of the same tissue type as the cancer.

Also described herein are methods for assaying a cancer for PD-L1 and then treating the cancer with an anti-PD-1 antibody, optionally in combination

- 53 040365 with one or more immunomodulators, such as an anti-TIM-3 antibody described herein, an anti-LAG-3 antibody, or an anti-PD-L1 antibody. As described in Example 10 herein, a cancer sample can be analyzed for protein levels or mRNA levels of PD-L1. A sample having levels of PD-L1 (protein or mRNA) greater than a reference value or a value in a control cell (eg, a non-cancerous cell) can be classified as PD-L1 positive. Thus, in some embodiments, an anti-PD-1 antibody (optionally in combination with one or more anti-cancer agents, optionally in combination with one or more immunomodulators such as an anti-TIM-3 antibody described herein, an anti-LAG -3 or anti-PD-L1 antibody) is administered to a patient who has or is identified as having a cancer that is positive for PD-L1. The malignant tumor may be, for example, non-small cell lung adenocarcinoma (NSCLC) (ASA), NSCLC squamous cell carcinoma (SCC), or hepatocellular carcinoma (HCC).

Based on, for example, example 9 of the present description, it was found that certain types of gastric cancer, which are triple positive for PDL1/CD8/IFN-y, are also positive for PIK3CA. Thus, in some embodiments, a cancer may be treated with an anti-PD-1 antibody molecule (optionally in combination with one or more immunomodulators, e.g., an anti-LAG-3 antibody molecule, an anti-TIM-3 antibody molecule as described herein, or a antibodies against PD-L1) and an agent that inhibits PIK3CA. Illustrative means 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):23748, 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 .

Based on, for example, Example 9 of the present disclosure, a CRC of, for example, a patient who has (or is identified as having) a high MSI CRC can be treated with an anti-PD-1 antibody, optionally in combination with a therapeutic agent that targets one or more of TIM-3, such as the anti-TIM-3 antibody described herein, LAG-3, RNF43 and BRAF. For example, these cancers can be treated with an anti-PD-1 antibody, optionally in combination with one or more therapeutic agents that target one or more of TIM-3, LAG-3, PD-1, RNF43, and BRAF. In embodiments, the one or more therapeutic agents include immunomodulators such as an anti-TIM-3 antibody described herein, an anti-LAG-3 antibody molecule, and an anti-cancer agent described in Table 1. 6 or in the publication given in table. 6. LAG-3 inhibitors, such as antibodies, are described herein. RNF43 can be inhibited, for example, by an antibody, a small molecule compound (for example, 2-(2',3-dimethyl-[2,4'-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridine -2-yl)acetamide (compound A28)), miRNA or Rspo ligand or derivative thereof. BRAF inhibitors (eg, vemurafenib or dabrafenib) are described herein.

Based on, for example, Example 9 of this disclosure, a patient who has (or is identified as having) squamous cell lung cancer can be treated with an anti-PD-1 antibody molecule in combination with a therapeutic agent that targets TIM-3, such as an anti-TIM-3 antibody molecule. 3, LAG-3, such as an anti-LAG-3 antibody molecule, and optionally one or more anti-cancer agents, such as the anti-cancer agent described in Table 3. 6 or in the publication given in table. 6.

In some embodiments, an individual who has (or is identified as having) squamous cell lung cancer can be treated with an anti-PD-1 antibody, optionally in combination with a therapeutic agent that targets TIM-3, such as an anti-TIM-3 antibody described herein. .

Based on, for example, Example 9 of the present disclosure, a patient who has (or is identified as having) thyroid cancer can be treated with an anti-PD-1 antibody molecule, optionally in combination with a therapeutic agent that targets BRAF, and optionally in combination with one or several immunomodulators, for example, an anti-LAG-3 antibody molecule, an anti-TIM-3 antibody molecule described herein, and an anti-PDL1 antibody molecule. BRAF inhibitors (eg, vemurafenib or dabrafenib) are described herein, eg in Table. 6 and in the publications given in table. 6.

In other embodiments, the cancer is a hematological cancer or cancer, including but not limited to leukemia or lymphoma. For example, an anti-TIM-3 antibody molecule can be used to treat malignant tumors and neoplasms, including, but not limited to, for example, acute leukemias, including, but not limited to, for example, B-cell acute lymphoid leukemia (BALL), T-cell acute lymphoid leukemia (TALL), acute lymphoid leukemia (ALL); one or more of chronic leukemias, including but not limited to, for example, chronic myelogenous leukemia (CML), chronic lymphocytic leukemia (CLL); additional hematologic malignancies

- 54 040365 natural tumors or hematological conditions, including, but not limited to, for example, B-cell prolymphocytic leukemia, blast plasmacytoid neoplasm of dendritic cells, Burkitt's lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell or large cell follicular lymphoma, malignant lymphoproliferative conditions, MALT lymphoma, mantle cell lymphoma, marginal zone cell lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndrome, non-Hodgkin's lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenström macroglobulinemia and preleukemia, which are diverse a set of hematological conditions united by ineffective production (or dysplasia) of myeloid blood cells, etc.

In some embodiments, an anti-TIM-3 antibody molecule is used to treat a cancer that expresses TIM-3. TIM-3 expressing malignancies include cervical cancer (Cao et al., PLoS One. 2013;8(1):e53834), lung cancer (Zhuang et al., Am J Clin Pathol. 2012;137(6):978 -985) (eg, non-small cell lung cancer), acute myeloid leukemia (Kikushige et al., Cell Stem Cell. 2010 Dec 3;7(6):708-17), diffuse large B-cell lymphoma, melanoma (Fourcade et al ., JEM, 2010; 207(10):2175), kidney cancer (e.g., renal cell carcinoma (RCC), e.g., clear cell carcinoma of the kidney, papillary cell carcinoma of the kidney, or metastatic renal cell carcinoma), squamous cell carcinoma, esophageal squamous cell carcinoma, nasopharyngeal carcinoma, colon and rectal cancer, breast cancer (eg, breast cancer that does not express one, two, or all of the estrogen receptor, progesterone receptor, or Her2/neu, eg, triple negative breast cancer), mesothelioma, hepatocellular carcinoma and ovarian cancer. The TIM-3 expressing cancer may be a metastatic cancer. In other embodiments, an anti-TIM-3 antibody molecule is used to treat a cancer that is characterized by macrophage activity or high expression of macrophage cell markers. In one embodiment, an anti-TIM-3 antibody molecule is used to treat a cancer that is characterized by high expression of one or more of the following macrophage cell markers: LILRB4 (macrophage inhibitory receptor), CD14, CD16, CD68, MSR1, SIGLEC1, TREM2, CD163, ITGAX, ITGAM, CD11b or CD11c. Examples of such malignancies include, but are not limited to, diffuse large B cell lymphoma, glioblastoma multiforme, clear cell carcinoma of the kidney, pancreatic adenocarcinoma, sarcoma, hepatocellular carcinoma of the liver, adenocarcinoma of the lung, renal papillary cell carcinoma, cutaneous melanoma, low-grade glioma brain, squamous cell carcinoma of the lung, serous cystadenocarcinoma of the ovary, squamous cell carcinoma of the head and neck, invasive breast carcinoma, acute myeloid leukemia, squamous cell carcinoma of the cervix, endocervical adenocarcinoma, uterine carcinoma, colon and rectal cancer, endometrial carcinoma of the uterine body, thyroid carcinoma gland, carcinoma of the urothelium of the bladder, carcinoma of the adrenal gland, chromophobe adenocarcinoma of the kidney and adenocarcinoma of the prostate.

In one embodiment, the cancer is lung cancer, such as lung adenocarcinoma.

In another embodiment, the cancer is a kidney cancer, such as renal cell carcinoma (RCC) (eg, clear cell carcinoma of the kidney or papillary cell carcinoma of the kidney) or a metastatic lesion thereof.

In another embodiment, the cancer is mesothelioma.

In another embodiment, the cancer is nasopharyngeal carcinoma (NPC).

In another embodiment, the cancer is a hematologic cancer (eg, myeloid leukemia, eg, acute myeloid leukemia (AML)).

In another embodiment, the cancer is a lymphoma (eg, diffuse large B-cell lymphoma).

In another embodiment, the cancer is breast cancer, eg, triple negative (TN) and/or immunomodulatory subtype.

In another embodiment, the cancer is glioblastoma multiforme.

In another embodiment, the cancer is ovarian cancer (eg, ovarian carcinoma).

In certain embodiments, the cancer is a solid tumor and the antibody molecule is administered in combination with an anti-LAG-3 or anti-PD-1 antibody molecule.

- 55 040365

Combination of Anti-TIM-3 Antibodies with Cancer Vaccines

Anti-TIM-3 antibody molecules can be combined with an immunogenic agent such as cancer cells, purified tumor antigens (including recombinant proteins, peptides 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 melanoma antigen peptides such as gp100 peptides, MAGE, Trp-2, MART1 and/or tyrosinase antigens, or tumor cells transfected to express the cytokine GM-CSF, DNA-based vaccines, RNA based vaccines; and viral transduction based vaccines. A cancer vaccine may be prophylactic or therapeutic.

In some embodiments, therapy with an anti-TIM-3 antibody molecule is combined with a vaccination protocol. Many experimental strategies have been developed for vaccination against tumors (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, see also 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, the vaccine is produced 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. GMCSF has been shown to be a potent antigen presentation activator for tumor vaccination (Dranoff et al. (1993) Proc. Natl. Acad. Sci. U.S.A. 90: 3539-43).

Anti-TIM-3 antibody molecules can be used in conjunction with a collection of recombinant tumor-expressed proteins and/or peptides to induce an immune response to these proteins. These proteins are normally considered by the immune system as self antigens and thus are tolerant to them. The tumor antigen may also include the protein telomerase, which is required for the synthesis of chromosome telomeres and which is expressed in more than 85% of malignant tumors and only in a limited number of somatic tissues (Kim, N et al. (1994) Science 266:2011-2013) (these somatic tissues can be protected from immune attack in various ways). Tumor antigens can also be neoantigens expressed in malignant cells due to somatic mutations that change the protein sequence or form fusion proteins between two unrelated sequences (eg, bcr-abl on the Philadelphia chromosome), or an idiotype in the case of B-cell tumors.

Other tumor vaccines may include proteins from viruses involved in human cancers such as human papillomaviruses (HPV), hepatitis viruses (HBV and HCV) and Kaposi's sarcoma herpes virus (KHSV) and Epstein-Barr virus (EBV). Another form of specific tumor antigen that can be used in conjunction with an anti-TIM-3 antibody is purified heat shock proteins (HSPs) isolated directly from tumor tissue. These heat shock proteins contain fragments of proteins from tumor cells and these HSPs are highly efficient in delivering 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).

Dendritic cells (DC) are efficient antigen-presenting cells that can be used to induce antigen-specific responses. DC can be produced ex vivo and loaded with various protein and peptide antigens as well as tumor cell extracts (Nestle, F. et al. (1998) Nature Medicine 4: 328-332). DCs can also be genetically transduced to express these tumor antigens. DCs have been fused directly to tumor cells for immunization purposes (Kugler, A. et al. (2000) Nature Medicine 6:332336). As a vaccination method, DC immunization can be effectively combined with anti-TIM-3 therapy to activate more potent antitumor responses.

Alternatively or in combination, the combination further comprises an immune response checkpoint modulator inhibitor or activator, e.g. LAG-3 inhibitor (e.g. anti-TIM-3 antibody molecule), PD-L1 inhibitor (e.g. anti-PD-L1 antibody molecule) , a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) or a CTLA-4 inhibitor (eg, an anti-CTLA4 antibody), or any combination thereof.

TIM-3 blockade can also be combined with standard cancer treatment. TIM-3 blockade can be effectively combined with chemotherapy regimens. In these cases, it may be possible to reduce the dose of the chemotherapeutic agent administered (Mokyr, M. et al. (1998) Cancer Research 58: 5301-5304). In certain embodiments, the methods and administration of the compositions described herein are carried out in combination with one or more other antibody molecules, chemotherapy, other cancer therapy (e.g., targeted cancer therapy or oncolytic drugs), cytotoxic agents, immune therapies (eg, cytokines), surgical and/or radiation procedures. Illustrative cytotoxic agents that can be administered

- 56 040365 combinations include antimicrotubulin agents, topoisomerase inhibitors, antimetabolites, mitosis inhibitors, alkylating agents, anthracyclines, vinca alkaloids, intercalating agents, agents capable of interfering with the signal transduction cascade, agents that stimulate apoptosis, proteasome inhibitors, and radiation therapy ( e.g. local irradiation or irradiation of the whole organism).

Alternatively, or in combination with the above combinations, the methods and administration of the compositions described herein may be carried out in combination with one or more of: an immunomodulator (eg, an activator of a costimulatory molecule or an inhibitor of an inhibitory molecule); vaccines, for example a therapeutic cancer vaccine; or other forms of cellular immunotherapy.

Illustrative non-limiting combinations and uses of anti-TIM-3 antibody molecules include the following combinations.

In certain embodiments, an anti-TIM-3 antibody molecule is administered in combination with a modulator of a co-stimulatory molecule or an inhibitory molecule, such as a co-inhibitory ligand or receptor.

In one embodiment, an anti-TIM-3 antibody molecule is administered in combination with a modulator, such as an agonist, of a co-stimulatory molecule. In one embodiment, the co-stimulatory molecule agonist is selected from an agonist (e.g., antibody agonist or antigen binding fragment thereof or soluble fusion construct) 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 ligand.

In another embodiment, an anti-TIM-3 antibody molecule is used in combination with a co-stimulatory molecule, eg, a positive signal-associated agonist, which includes the co-stimulatory domain of CD28, CD27, ICOS, and GITR.

Exemplary GITR agonists include, for example, GITR fusion proteins and anti-GITR antibodies (e.g., bivalent anti-GITR antibodies) such as the GITR fusion protein described in US Pat. No. 6,111,090, EP No. 090505B1, US Pat. 2010/003118 and 2011/090754, or an anti-GITR antibody as described, for example, in US Patent No. 7025962, EP No. 1947183B1, US Patent No. 7812135, US Patent No. 8388967, US Patent No. 8591886, EP Patent No. 1866339, publications PCT Publication No. WO 2011/028683, PCT Publication No. WO 2013/039954, PCT Publication No. WO 2005/007190, PCT Publication No. WO 2007/133822, PCT Publication No. WO 2005/055808, PCT Publication No. WO 99/40196, PCT Publication No. WO 2001/03720, PCT Publication No. WO 99/20758, PCT Publication No. WO 2006/083289, PCT Publication No. WO 2005/115451, US Patent No. 7618632 and PCT Publication No. WO 2011/051726. In one embodiment, the anti-TIM-3 antibody molecule is administered in combination with an immune response checkpoint inhibitor (or immune inhibitor molecule). The term immune checkpoints, as used herein, refers to a group of molecules on the cell surface of immune cells, such as CD4 and CD8 T cells, that serve as brakes, suppressing or inhibiting an immune response, such as an antitumor immune response. Immune response checkpoint molecules include, but are not limited to, programmed death protein 1 (PD-1), PD-L1, cytotoxic T-lymphocyte antigen 4 (CTLA-4), B7-H1, B7-H3, B7-H4, OX-40, 41BB (CD137), CD40, T-cell immunoglobulin domain and mucin domain 3 (TIM-3), and lymphocyte activation gene 3 (LAG-3), among others.

Immunotherapeutic agents that may act as inhibitors of immune response checkpoint molecules useful in combination with the anti-PD-1 antibody molecules described herein include, but are not limited to, PD-L1, PD-L2, CTLA-4 inhibitors. , TIM-3, LAG-3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4, CEACAM (eg CEACAM-1, CEACM-3 and/or CEACAM-5) and/or TGFR-beta. Inhibition of an immune inhibitory molecule can be carried out by inhibition at the DNA, RNA or protein level. In embodiments, an inhibitory nucleic acid (eg, dsRNA, siRNA, or shRNA) can be used to inhibit the expression of an inhibitory molecule. In other embodiments, the inhibitor of the inhibitory signal is a polypeptide, such as a soluble ligand or an antibody or antigen-binding fragment thereof, that binds to an inhibitory molecule.

In one embodiment, the inhibitor is a soluble ligand (eg, CTLA-4-Ig or TIM-3-Ig) or an antibody or antibody fragment that binds to CTLA-4. For example, an anti-TIM-3 antibody molecule can be administered in combination with an anti-CTLA-4 antibody, such as ipilimumab, for example, to treat a cancer (eg, a cancer selected from: melanoma, such as metastatic melanoma; lung cancer, such as non-small cell lung carcinoma or prostate cancer). Exemplary anti-CTLA-4 antibodies include tremelimumab (an IgG2 monoclonal antibody available from Pfizer, formerly known as ticilimumab, CP-675.206); and ipilimumab (an anti-CTLA-4 antibody also known as MDX010, CAS no. 477202-00-9). In one embodiment, the anti-TIM-3 antibody molecule is administered

- 57 040365 after treatment, eg after treatment of melanoma, with an anti-CTLA-4 antibody (eg ipilimumab) with or without a BRAF inhibitor (eg vemurafenib or dabrafenib). Exemplary doses that may be used include an anti-TIM-3 antibody dose of about 1 to 30 mg/kg, 1 to 20 mg/kg, or 1 to 10 mg/kg, such as 3 mg/kg, and an anti-CTLA antibody dose. -4, such as ipilimumab, approximately 3 mg/kg.

In certain embodiments, immune response checkpoint molecules, eg, PD-1, LAG-3, TIM-3, CEACAM-1/-5, can regulate T cell function, accelerating tumor immune escape. Thus, the anti-TIM-3 antibodies described herein can be used in combination with one or more inhibitors of these immune inhibitory molecules to enhance an antitumor response. The combination of antibodies described herein can be administered singly, eg as single antibodies, or linked, eg as a bispecific or trispecific antibody molecule.

In one embodiment, an anti-TIM-3 antibody molecule is administered in combination with an anti-TIM-3 antibody or antigen-binding fragment thereof. In another embodiment, an anti-TIM-3 antibody molecule is administered in combination with an anti-PD-1 antibody or antigen-binding fragment thereof. In other embodiments, an anti-TIM-3 antibody molecule is administered in combination with an anti-TIM-3 antibody and an anti-PD-1 antibody or anigen-binding fragments thereof. In one embodiment, a bispecific antibody is administered that comprises an anti-TIM-3 antibody molecule and an anti-PD-1 or anti-TIM-3 antibody or antigen-binding fragment thereof. In certain embodiments, a combination of antibodies described herein is used to treat a cancer, eg, a cancer as described herein (eg, a solid tumor). The efficacy of the above combinations can be tested in animal models known in the art. For example, animal models for studying the effect against PD-1 and against LAG-3 are described, for example, in Woo et al. (2012) Cancer Res. 72(4):917-27).

In some embodiments, inhibitors of TIM-3 and PD-1 molecules (eg, anti-TIM-3 and anti-PD-1 antibody molecules) are administered in combination, for example, to treat cancer. In some embodiments, the subject is a patient who has progressed (e.g., tumor growth) during therapy with a PD-1 inhibitor (e.g., an antibody molecule as described herein) and/or a PD-L1 inhibitor (e.g., a molecule antibodies against PD-L1). In some embodiments, therapy with an anti-PD-1 antibody molecule and/or an anti-PD-L1 antibody molecule is continued and an immune TIM-3 inhibitory molecule (eg, antibody) is added to the therapy.

In other embodiments, the anti-TIM-3 antibody molecule is administered in combination with a CEACAM inhibitor (eg, an inhibitor of CEACAM-1, CEACAM-3, and/or CEACAM-5). In one embodiment, the CEACAM inhibitor is an anti-CEACAM antibody molecule. In one embodiment, an anti-TIM-3 antibody molecule is administered in combination with a CEACAM-1 inhibitor, such as an anti-CEACAM-1 antibody molecule. In another embodiment, an anti-TIM-3 antibody molecule is administered in combination with a CEACAM-3 inhibitor, such as an anti-CEACAM-3 antibody molecule. In another embodiment, an anti-TIM-3 antibody molecule is administered in combination with a CEACAM-5 inhibitor, eg, an anti-CEACAM-5 antibody molecule. Illustrative antibodies against CEACAM-1 are described in WO 2010/125571, WO 2013/082366 and WO 2014/022332, for example, monoclonal antibody 34B1, 26H7 and 5F4; or their recombinant form, as described, for example, in US 2004/0047858, US 7132255 and WO99/052552. In other embodiments, the anti-CEACAM antibody binds to CEACAM-5 as described, for example, in Zheng et al. PLOS One. 2010 Sep 2;5(9). pii: e12529 (DOI:10:1371/journal.pone.0021146) or cross-reacts with CEACAM-1 and CEaCaM-5 as described for example in WO 2013/054331 and US 2014/0271618.

Without wishing to be bound by theory, it is believed that the carcinoembryonic antigen cell adhesion molecule (CEACAM), such as CEACAM-1 and CEACAM-5, mediates at least in part the inhibition of the antitumor immune response (see, e.g., Markel et al. J Immunol. 2002 Mar 15;168(6):280310 Markel et al J Immunol 2006 Nov 1;177(9):6062-71 Markel et al Immunology 2009 Feb 126(2):186-200 Markel et al Cancer Immunol Immunother 2010 Feb 59(2):215-30 Ortenberg et al Mol Cancer Ther 2012 Jun 11(6):1300-10 Stern et al J Immunol 2005 Jun 1; 174(11):6692-701 Zheng et al PLoS One 2010 Sep 2 5(9) pii:e12529). For example, CEACAM-1 has been described as a heterophilic ligand for TIM-3 and as playing a role in TIM-3 mediated T-cell tolerance and depletion (see e.g. WO 2014/022332; Huang, et al. (2014) Nature doi:10.1038/naturel3848). In embodiments, co-blockade of CEACAM-1 and TIM-3 has been shown to enhance the antitumor effect in colon cancer xenograft models (see e.g. WO 2014/022332; Huang, et al. (2014), supra) . In other embodiments, the implementation of the joint blockade of CEACAM-1 and PD-1 reduces T-cell tolerance, as described, for example, in WO 2014/059251. Thus, CEACAM inhibitors can be used with other immunomodulators described herein (e.g., PD-1 and/or TIM-3 inhibitors) to enhance the immune response against cancer, e.g., melanoma, lung cancer (e.g., NSCLC), urinary cancer. bubble, thick cancer

- 58 040365 th intestine of ovarian cancer and other malignant tumors, as described in the present description.

In some embodiments, immune inhibitory PD-1 and TIM-3 molecules (eg, antibody molecules) are administered in combination with each other, for example, to treat cancer. In some embodiments, the patient is a patient who has progressed (e.g., tumor growth has occurred) during therapy with a PD-1 inhibitor (e.g., an antibody molecule as described herein) and/or a PDL1 inhibitor (e.g., an antibody molecule) . In some embodiments, therapy with an anti-PD-1 antibody molecule and/or an anti-PDL1 antibody molecule is continued and an immune TIM-3 inhibitory molecule (eg, antibody) is added to the therapy.

In some embodiments, immune inhibitory TIM-3 and LAG-3 molecules (eg, antibody molecules) are administered in combination with each other, for example, to treat cancer. In some embodiments, the patient is a patient who progressed (e.g., tumor growth) during therapy with a TIM-3 inhibitor (e.g., an antibody molecule as described herein) and/or a PD-1 inhibitor (e.g., a antibodies). In some embodiments, therapy with an anti-TIM-3 antibody molecule and/or an anti-PDL1 antibody molecule is continued and an immune LAG-3 inhibitory molecule (eg, antibody) is added to the therapy.

In other embodiments, the anti-TIM-3 antibody molecule is administered in combination with a cytokine, such as interleukin-21, interleukin-2, or interleukin-15. In certain embodiments, the combination of an anti-TIM-3 antibody molecule and a cytokine described herein is used to treat a cancer, eg, a cancer as described herein (eg, a solid tumor or melanoma).

Illustrative immunomodulators that can be used in combination with anti-TIM-3 antibody molecules include, but are not limited to, for example, afutuzumab (available from Roche®); pegfilgrastim (Neulasta®); lenalidomide (CC-5013, Revlimid®); thalidomide (Thalomid®), actimide (CC4047) and cytokines such as IL-21 or IRX-2 (a mixture of human cytokines including interleukin 1, interleukin 2 and interferon y, CAS 951209-71-5, available from IRX Therapeutics).

In other embodiments, an anti-TIM-3 antibody molecule is used in combination with an indolamine-pyrrole-2,3-dioxygenase (IDO) inhibitor (e.g., INCB24360) in an individual with advanced or metastatic cancer (e.g., in a patient with metastatic or recurrent cancer). NSCL tumor).

In other embodiments, anti-TIM-3 antibody molecules are co-administered to an individual with (e.g., before, concurrently, or after) one or more of: bone marrow transplantation, T-cell depleting therapy using chemotherapeutic agents such as fludarabine, external beam radiation therapy (XRT), cyclophosphamide and/or antibodies such as OKT3 or CAMPATH. In one embodiment, anti-TIM-3 antibody molecules are administered following B-cell depleting therapy, such as agents that react with CD20, such as Rituxan. For example, in one embodiment, individuals may be treated with standard high dose chemotherapy followed by peripheral blood stem cell transplantation. In certain embodiments, anti-TIM-3 antibody molecules are administered to individuals after transplantation. In a further embodiment, anti-TIM-3 antibody molecules are administered before or after surgery.

Another example of a combination is an anti-TIM-3 antibody in combination with decarbazine for the treatment of melanoma. Without being bound by theory, it is believed that the combined use of TIM-3 blockade and chemotherapy is promoted by cell death, which is a consequence of the cytotoxic effect of most chemotherapeutic compounds, which can lead to increased levels of tumor antigen in the antigen presentation cascade. Other combination therapies that may result in synergy with TIM-3 blockade through cell death are radiotherapy, surgery, and hormone deficiency. Each of these protocols creates a source of tumor antigen in the host. Angiogenesis inhibitors can also be combined with TIM-3 blockade. Inhibition of angiogenesis results in the death of tumor cells, which can provide tumor antigen to host antigen presentation cascades.

TIM-3 blocking antibodies can also be used in combination with bispecific antibodies. Bispecific antibodies can be used to target two separate antigens. For example, anti-Fc receptor/anti-tumor antigen bispecific antibodies (eg, Her-2/neu) are used to target macrophages to the tumor site. This targeting may more effectively activate tumor-specific responses. The T-cell link of these responses can be enhanced using TIM-3 blockade. Alternatively, the antigen can be delivered directly to the DC using bispecific antibodies that bind to the tumor antigen and a specific dendritic cell surface marker.

Tumors elude the host's immune surveillance through a wide variety of mechanisms. Many of these mechanisms can be overcome through the inactivation of proteins that are ex

- 59 040365 are pressed by tumors and which are immunosuppressive. These include, among others, TGF-beta (Kehrl, J. et al. (1986) J. Exp. Med. 163: 1037-1050), IL-10 (Howard, M. & O'Garra, A. (1992 ) Immunology Today 13: 198-200) and Fas ligand (Hahne, M. et al. (1996) Science 274: 1363-1365). Antibodies or antigen-binding fragments thereof to each of these structures can be used in combination with anti-TIM-3 antibody molecules to counteract the effects of an immunosuppressive agent and promote host immune responses.

Other antibodies that can be used to activate the immune response capability of the host can be used in combination with anti-TIM-3 antibody molecules. They include molecules on the surface of dendritic cells that activate DC function and antigen presentation. Anti-CD40 antibodies are able to effectively replace T cell helper activity (Ridge, J. et al. (1998) Nature 393: 474-478) and can be used in conjunction with anti-PD-1 antibodies (Ito, N. et al. ( 2000) Immunobiology 201 (5) 527-40). Antibodies to T-cell co-stimulatory molecules such as CTLA-4 (e.g. US Pat. No. 5,811,097), OX-40 (Weinberg, A. et al. (2000) Immunol 164: 2160-2169), 4-1BB (Melero, I. et al. (1997) Nature Medicine 3: 682-685 (1997) and ICOS (Hutloff, A. et al. (1999) Nature 397: 262-266) can also provide increased levels of T cell activation.

Additional exemplary standard treatments are described in the section entitled Combination Therapies below.

In all of the methods described herein, TIM-3 blockade can be combined with other forms of immunotherapy, such as cytokine treatment (eg, interferons, GM-CSF, G-CSF, IL-2, IL-21) or bispecific antibody therapy. that provide enhanced presentation of tumor antigens (see, for example, Holliger (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak (1994) Structure 2:1121-1123).

Methods for introducing antibody molecules are known in the art and are described below. Suitable dosages of the molecules used depend on the age and body weight of the individual and the particular drug used. Dosages and therapeutic regimens of the anti-TIM3 antibody molecule can be determined by the skilled artisan. In certain embodiments, the anti-TIM-3 antibody molecule is administered by injection (e.g., subcutaneously or intravenously) at a dose of about 1 to 30 mg/kg, e.g., about 5 to 25 mg/kg, about 10 to 20 mg/kg, about 1 to 5 mg/kg, or about 3 mg/kg. In some embodiments, the anti-TIM-3 antibody molecule is administered at a dose of about 1 mg/kg, about 3 mg/kg, about 10 mg/kg, about 15 mg/kg, about 20 mg/kg, about 25 mg/kg, or about 30 mg/kg. In some embodiments, the anti-TIM-3 antibody molecule is administered at a dose of about 1-3 mg/kg, about 3-10 mg/kg, about 3-15 mg/kg, about 10-15 mg/kg, about 10-20 mg /kg, approximately 10-25 mg/kg or approximately 20-30 mg/kg. In some embodiments, the anti-TIM-3 antibody molecule is administered at a dose of about 0.5-2, 2-4, 2-5, or 5-15 mg/kg. The dosing schedule may vary, for example, from once a week to once every 2, 3 or 4 weeks. In one embodiment, the anti-TIM-3 antibody molecule is administered at a dose of about 10 to 20 mg/kg biweekly.

The antibody molecules can be used alone or in conjugated form with a second agent, such as a cytotoxic drug, a radioactive isotope, or a protein, such as a protein toxin or a viral protein. This method includes: administering an antibody molecule, alone or conjugated to a cytotoxic drug, to an individual in need of such treatment. Antibody molecules can be used to deliver various drugs, e.g., a cytotoxic moiety, e.g., a therapeutic drug, a radioactive isotope, molecules of plant, fungal, or bacterial origin, or biological proteins (e.g., protein toxins) or particles (e.g., recombinant viral particles, e.g. ; through the viral envelope protein) or mixtures thereof.

Anti-TIM-3 antibody molecules can also be combined with standard cancer treatments. For example, anti-TIM-3 antibody molecules can be effectively combined with chemotherapy regimens. In these cases, it may be possible to reduce the dose of the injected chemotherapeutic agent (Mokyr, M. et al. (1998) Cancer Research 58:301-5304). An example of such a combination is an anti-TIM-3 antibody molecule in combination with decarbazine for the treatment of melanoma. Another example of such a combination is an anti-TIM-3 antibody molecule in combination with interleukin-2 (IL-2) for the treatment of melanoma. In some embodiments, an anti-TIM-3 antibody molecule can be combined with IL-21. Without being bound by theory, one scientific rationale for the combined use of anti-TIM-3 antibody molecule therapy and chemotherapy is that cell death, which is a consequence of the cytotoxic effect of most chemotherapeutic compounds, should lead to increased levels of tumor antigen in the antigen presentation cascade. Other combination therapies that can result in synergy with anti-TIM-3 antibody molecule therapy through cell death are radiotherapy, surgery, and hormone deficiency. Each of these protocols creates a source of tumor antigen in the host. Also, anti-TIM-3 antibody molecule therapy can be combined

- 60 040365 angiogenesis inhibitors. Inhibition of angiogenesis results in the death of tumor cells, which can provide tumor antigen to host antigen presentation cascades. Anti-TIM-3 antibody molecules can also be used in combination with bispecific antibodies. Bispecific antibodies can be used to target two separate antigens. For example, anti-Fc receptor/anti-tumor antigen bispecific antibodies (eg, Her2/neu) are used to target macrophages in the tumor region. This targeting may more effectively activate tumor-specific responses. The T-cell link of these responses can be enhanced by the use of anti-TIM-3 antibody molecules. Alternatively, the antigen can be delivered directly to the DC using bispecific antibodies that bind to the tumor antigen and a specific dendritic cell surface marker.

Tumors elude the host's immune surveillance through a wide variety of mechanisms. Many of these mechanisms can be overcome by inactivating proteins that are expressed by tumors and that are immunosuppressive. These include, among others, TGF-beta (Kehrl, J. et al. (1986) J. Exp. Med. 163: 1037-1050), IL-10 (Howard, M. & O'Garra, A. (1992 ) Immunology Today 13: 198-200), and Fas ligand (Hahne, M. et al. (1996) Science 274: 1363-1365). Antibodies to each of these structures can be used in combination with anti-TIM-3 antibody molecules to counteract the effects of an immunosuppressive agent and promote host immune responses.

In combination with anti-TIM-3 antibody molecules, other antibodies can be used that can be used to activate the host's ability to mount an immune response. They include molecules on the surface of dendritic cells that activate DC function and antigen presentation. Anti-CD40 antibodies are able to effectively replace T cell helper activity (Ridge, J. et al. (1998) Nature 393: 474-478) and can be used in conjunction with anti-PD-1 antibodies (Ito, N. et al. ( 2000) Immunobiology 201 (5) 527-40). Activating antibodies to T-cell costimulatory molecules such as CTLA-4 (e.g. US Pat. No. 5,811,097), OX-40 (Weinberg, A. et al. (2000) Immunol 164: 2160-2169), 4-1BB (Melero , I. et al. (1997) Nature Medicine 3:682-685 (1997) and ICOS (Hutloff, A. et al. (1999) Nature 397:262-266) can also provide increased levels of T cell activation.

Additional Combination Therapies

The anti-TIM-3 antibody molecule can be used in combination with other therapies. For example, combination therapy may include a composition of the present invention co-formulated and/or co-administered with one or more additional drugs, such as one or more anti-cancer agents, cytotoxic or cytostatic agents, hormonal treatments, vaccines, and/or other immunotherapies. . In other embodiments, the antibody molecules are administered in combination with other therapies, including surgery, radiation therapy, cryosurgery, and/or thermotherapy. Such combination therapies can advantageously use lower dosages of administered drugs, thus avoiding the potential toxicity or complications associated with various monotherapies.

The term in combination with means that the administration of therapy or administration of drugs must be carried out at the same time and / or be combined for delivery, although these methods of delivery are included in the scope described in the present description. Molecules of antibodies against TIM-3 can be administered simultaneously, before or after one or more other additional therapies or drugs. Administration of the anti-TIM-3 antibody molecule and other agent, or administration of the therapeutic protocol, can be done in any order. Typically, each agent is administered at a dose and/or schedule determined for that agent. In addition, it will be understood that the additional drug used in this combination can be administered together in one composition or administered separately in different compositions. In general, it is expected that additional drugs used in combination can be used at levels that do not exceed the levels at which they are used separately. In some embodiments, levels used in combination will be lower than levels used alone.

In certain embodiments, the anti-TIM-3 antibody molecules described herein are administered in combination with one or more other inhibitors of TIM-3 or other immune response checkpoint molecules, e.g., PD-1, PD-L1, PD-L2, CEACAM (eg CEACAM-1, CEACAM-3 or CEACAM-5) or LAG-3.

In certain embodiments, the anti-TIM-3 antibody molecules described herein are administered in combination with one or more other PD-1, PD-L1 and/or PD-L2 inhibitors known in the art. The antagonist may be an antibody, an antigen-binding fragment thereof, an immunoadhesin, a fusion protein, or an oligopeptide. In some embodiments, the anti-PD-1 antibody is selected from MDX-1106, Merck 3475, or CT-011. In some embodiments, the PD-1 inhibitor is an immunoadhesin (e.g., an immunoadhesin comprising an extracellular or PD-1 binding portion of PD-L1 or PD-L2 fused to a constant region

- 61 040365 (for example, the Fc region of an immunoglobulin sequence)). In some embodiments, the PD-1 inhibitor is AMP-224. In some embodiments, the PD-L1 inhibitor is an anti-PD-L1 antibody. In some embodiments, the PD-L1 binding antagonist is selected from YW243.55.S70, MPDL3280A, MEDI-4736, MSB-0010718C, or MDX-1105. MDX-1105, also known as BMS-936559, is an anti-PD-L1 antibody described in WO 2007/005874. The YW243.55.S70 antibody (heavy and light chain variable region sequences shown in SEQ ID NOs: 20 and 21, respectively) is an anti-PD-L1 antibody described in WO 2010/077634.

MDX-1106, also known as MDX-1106-04, ONO-4538 or BMS-936558, is an anti-PD-1 antibody described in WO 2006/121168. Merck 3745, also known as MK-3475 or SCH-900475, is an anti-PD-1 antibody described in WO 2009/114335. Pidilizumab (CT-011; Cure Tech) is a humanized IgG1k monoclonal antibody that binds to PD-1. Pidilizumab and other humanized anti-PD-1 monoclonal antibodies are described in WO 2009/101611. In other embodiments, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab (trade name Keytruda, formerly lambrolizumab, also known as MK-3475) is described, for example, in Hamid, O. et al. (2013) New England Journal of Medicine 369(2):134-44. AMP-224 (B7-DCIg; Amplimmune; e.g. described in WO 2010/027827 and WO 2011/066342) is a soluble PD-L2 receptor Fc fusion that blocks the interaction between PD-1 and B7-H1. Other anti-PD-1 antibodies include AMP 514 (Amplimmune), among others, for example the anti-PD-1 antibodies described in US 8609089, US 2010028330 and/or US 20120114649.

In some embodiments, the anti-PD-1 antibody is MDX-1106. Alternative names for MDX-1106 include MDX-1106-04, ONO-4538, BMS-936558, or nivolumab. In some embodiments, the anti-PD-1 antibody is nivolumab (CAS accession number: 946414-94-4). Nivolumab (also referred to as BMS-93 6558 or MDX1106; Bristol-Myers Squibb) is a fully human IgG4 monoclonal antibody that specifically blocks PD-1. Nivolumab (clone 5C4) and other human monoclonal antibodies that specifically bind to PD-1 are described in US 8008449 and WO 2006/121168. Pembrolizumab (trade name Keytruda, formerly labrolizumab, also known as MK-3475; Merck) is a humanized IgG4 monoclonal antibody that binds to PD-1. Lambrolizumab and other humanized anti-PD-1 antibodies are described in US 8354509 and WO 2009/114335. MDPL3280A (Genentech/Roche) is a human Fc optimized monoclonal IgG1 antibody that binds to PD-L1. MDPL3280A and other human monoclonal antibodies to PD-L1 are described in US Pat. No. 7,943,743 and US Publication No. 20120039906. Other PD-L1 binding compounds include YW243.55.S70 (heavy and light chain variable regions shown in SEQ ID NO: 20 and 21 in WO 2010/077634) and MDX-1105 (also referred to as BMS-936559, and for example the PD-L1 binding compounds described in WO 2007/005874).

Methods for the treatment of a malignant tumor

Exemplary combinations of anti-TIM-3 antibody molecules (alone or in combination with other stimulants) and standard of care for cancer include at least the following.

In certain embodiments, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule described herein, is used in combination with a standard cancer treatment that is a chemotherapeutic agent, including, but not limited to, anastrozole (Arimidex® ), bicalutamide (Casodex®), bleomycin sulfate (Blenoxane®), busulfan (Myleran®), busulfan injectable (Busulfex®), capecitabine (Xeloda®), N4-pentoxycarbonyl-5-deoxy-5-fluorocytidine, carboplatin (Paraplatin® ), carmustine (BiCNU®), chlorambucil (Leukeran®), cisplatin (Platinol®), cladribine (Leustatin®), cyclophosphamide (Cytoxan® or Neosar®), cytarabine, cytosine arabinoside (Cytosar-U®), cytarabine liposomal injection ( DepoCyt®), Dacarbazine (DTIC-Dome®), Dactinomycin (Actinomycin D, Cosmegane), Daunorubicin Hydrochloride (Cerubidine®), Daunorubicin Citrate Liposomal Injection (DaunoXome®), Dexamethasone, Docetaxel (Taxotere®), Doxorubicin Hydrochloride (Adriam ycin®, Rubex®), etoposide (Vepesid®), fludarabine phosphate (Fludara®), 5-fluorouracil (Adrucil®, Efudex®), flutamide (Eulexin®), tezacitibine, gemcitabine (difluorodeoxycytidine), hydroxyurea (Hydrea®), idarubicin (Idamycin®), ifosfamide (IFEX®), irinotecan (Camptosar®), L-asparaginase (ELSPAR®), leucovorin calcium, melphalan (Alkeran®), 6-mercaptopurine (Purinthol®), methotrexate (Folex®), mitoxantrone (Novantrone®), mylotarg, paclitaxel (Taxol®), phoenix (yttrium 90/MX-DTPA), pentostatin, polyfeprosan 20 implant with carmustine (Gliadel®), tamoxifen citrate (Nolvadex®), teniposide (Vumon®), 6thioguanine, thiotepa, tirapazamine (Tirazone®), topotecan hydrochloride injection (Hycamptin®), vinblastine (Velban®), vincristine (Oncovin®), and vinorelbine (Navelbine®), ibrutinib, idelicib, and brentuximab vedotin.

- 62 040365

Exemplary alkylating agents include, but are not limited to, nitrogen mustards, ethyleneimine derivatives, alkylsulfonates, nitrosoureas, and triazenes: uracil mustard®, Chlorethaminacil®, Demethyldopan®, Desmethyldopan®, Haemanthamine®, Nordopan®, Uracil nitrogen mustard®, Uracillost ®, Uracilmostaza®, Uramustin®, Uramustine®), chlormethine (Mustargen®), cyclophosphamide (Cytoxan®, Neosar®, Clafen®, Endoxan®, Procytox®, Revimmune™), ifosfamide (Mitoxana®), melphalan (Alkeran®) , chlorambucil (Leukeran®), pipobroman (Amedel®, Vercyte®), triethylenemelamine (Hemel®, Hexalen®, Hexastat®), triethylenethiophosphoramine, temozolomide (Temodar®), thiotepa (Thioplex®), busulfan (Busilvex®, Myleran®) , carmustine (BiCNU®), lomustine (CeeNU®), streptozocin (Zanosar®), and dacarbazine (DTIC-Dome®). Additional exemplary alkylating agents include, but are not limited to, oxaliplatin (Eloxatin®); temozolomide (Temodar® and Temodal®); dactinomycin (also known as actinomycin D, Cosmegen®); melphalan (also known as L-PAM, L-sarcolysin and phenylalanine mustard, Alkeran®); altretamine (also known as hexamethylmelamine (HMM), Hexalen®); carmustine (BiCNU®); bendamustine (Treanda®); busulfan (Busulfex® and Myleran®); carboplatin (Paraplatin®); lomustine (also known as CCNU, CeeNU®); cisplatin (also known as CDDP, Platinol® and Platinol®-AQ); chlorambucil (Leukeran®); cyclophosphamide (Cytoxan® and Neosar®); dacarbazine (also known as DTIC, DIC and imidazole carboxamide, DTIC-Dome®); altretamine (also known as hexamethylmelamine (HMM), Hexalen®); Ifosfamide (Ifex®); prenumustine; procarbazine (Matulane®); mechlorethamine (also known as nitrogen mustard, mustine and mechlorethamine hydrochloride, Mustargen®); streptazocin (Zanosar®); thiotepa (also known as thiophosphoamide, TESPA and TSPA, Thioplex®); cyclophosphamide (Endoxan®, Cytoxan®, Neosar®, Procytox®, Revimmune®); and bendamustine HCl (Treanda®).

Illustrative anthracyclines include, for example, doxorubicin (Adriamycin® and Rubex®); bleomycin (lenoxane®); daunorubicin (daunorubicin hydrochloride, daunomycin and rubidomycin hydrochloride, Cerubidine®); liposomal daunorubicin (daunorubicin citrate liposomes, DaunoXome®); mitoxantrone (DHAD, Novantrone®); epirubicin (Ellence™); idarubicin (Idamycin®, Idamycin PFS®); mitomycin C (Mutamycin®); geldanamycin; herbimycin; ravidomycin and deacetylravidomycin.

Exemplary vinca alkaloids that can be used in combination with anti-TIM-3 antibody molecules, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-L1, or anti-PD-1 antibody molecule) include, but are not limited to , vinorelbine tartrate (Navelbine®), vincristine (Oncovin®) and vindesine (Eldisine®)); vinblastine (also known as vinblastine sulfate, vincaleukoblastin and VLB, Alkaban-AQ® and Velban®); and vinorelbine (Navelbine®).

Exemplary proteasome inhibitors that can be used in combination with anti-PD-1 antibody molecules, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD-L1, or anti-TIM-3 antibody molecule) include, but are not limited to imi, bortezomib (Velcade®); carfilzomib (PX-171-007, (S)-4-methyl-N-((S)-1-(((S)-4-methyl-1-((R)-2-methyloxiran2-yl)-1 -oxopentan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)-2-((S)-2-(2-morpholinoacetamido)-4phenylbutanamido)pentanamide); marizomib (NPI-0052); ixazomib citrate (MLN-9708); delanzomib (CEP-18770); and O-methyl-N-[(2-methyl-5-thiazolyl)carbonyl]-L-seryl-O-methyl-N-[(1S)-2-[(2R)-2-methyl-2oxiranyl]-2 -oxo-1-(phenylmethyl)ethyl]-L-serinamide (ONX-0912).

In some embodiments, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule described herein, is used alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PD antibody molecule). -L1) in combination with a tyrosine kinase inhibitor (eg, a receptor tyrosine kinase (RTK) inhibitor). An exemplary tyrosine kinase inhibitor includes, but is not limited to, an inhibitor of the epidermal growth factor (EGF) cascade (e.g., an inhibitor of the epidermal growth factor receptor (EGFR)), an inhibitor of the vascular endothelial growth factor (VEGF) cascade (e.g., an inhibitor of the vascular endothelial factor receptor (VEGFR) (eg, VEGFR-1 inhibitor, VEGFR-2 inhibitor, VEGFR-3 inhibitor)), platelet-derived growth factor cascade (PDGF) inhibitor (eg, platelet-derived growth factor receptor (PDGFR) inhibitor (eg, PDGFRβ inhibitor)) , RAF-1 inhibitor, KIT inhibitor, and RET inhibitor. In some embodiments, the cancer agent used in combination with the hedgehog inhibitor is selected from the group consisting of: axitinib (AG013736), bosutinib (SKI-606), cediranib (RECENTIN™, AZD2171), dasatinib (SPRYCEL®, BMS- 354825), erlotinib (TARCEVA®), gefitinib (IRESSA®), imatinib (Gleevec®, CGP57148B, STI-571), lapatinib (TYKERB®, TYVERB®), lestaurtinib (CEP-701), neratinib (HKI-272), nilotinib (TASIGNA®), semaxanib (semaxinib, SU5416), sunitinib (SUTENT®, SU11248), toceranib (PALLADIA®), vandetanib (ZACTIMA®, ZD6474), vatalanib (PTK787, PTK/ZK), trastuzumab (HERCEPTIN®), bevacizumab (AVASTIN®), rituximab (RITUXAN®), cetuximab (ERBITUX®), panitumumab (VECTIBIX®), ranibizumab (Lucentis®), ni- 63 040365 lotinib (TASIGNA®), sorafenib (NEXAVAR®), alemtuzumab (CAMPATH® ), gemtuzumab ozagomycin (MYLOTARG®), ENMD-2076, PCI-32765, AC220, dovitinib lactate (TKI258, CHIR-258), BIBW 2992 (TOVOK™), SGX523, PF-04217903, PF-023410 66, PF-299804, BMS-777607, ABT-869, MP470, BIBF1120 (VARGATEF®), AP24534, JNJ-26483327, MGCD265, DCC-2036, BMS-690154, CEP-11981, tivozaniba (AV-951), OSI -930, MM-121, XL-184, XL-647, XL228, AEE788, AG-490, AST-6, BMS-599626, CUDC-101, PD153035, Pelitinib (EKB-569), Vandetanib (Zactima), WZ3146 , WZ4002, WZ8040, ABT869 (linifanib), AEE788, AP24534 (ponatinib), AV-951 (tivozanib), axitinib, BAY 73-4506 (regorafenib), bivanib alaninate (BMS-582664), brivanib (BMS-540215), cediranib (AZD2171), ChIR258 (dovitinib), CP673451, CYC116, E7080, Ki8751, masitinib (AB1010), MGcD-265, motesanib diphosphate (AMG-706), MP-470, OSI-930, pazopanib hydrochloride, PD173074, sorafenib tosylate ( Bay 43-9006), SU 5402, TSU-68 (SU6668), Watalaniba, XL880 (GSK1363089, EXEL-2880). Individual tyrosine kinase inhibitors are selected from sunitinib, erlotinib, gefitinib, or sorafenib.

In certain embodiments, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule described herein, is used alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PDL1 antibody molecule). ), in combination with vascular endothelial growth factor (VEGF) receptor inhibitors, including, but not limited to, bevacizumab (Avastin®), axitinib (Inlyta®); brivaniba alaninate (BMS582664, (S)-((R)-1 -(4-(4-Fluoro-2-methyl-1 H-indol-5-yloxy)-5-methylpyrrolo[2,1-f][1 ,2,4]triazin-6-yloxy)propan-2-yl)2-aminopropanoate); sorafenib (Nexavar®); pazopanib (Votrient®); sunitinib malate (Sutent®); cediranib (AZD2171, CAS 288383-20-1); vargatef (BIBF1120, CAS 928326-83-4); foretinib (GSK1363089); telatinib (BAY57-9352, CAS 332012-40-5); apatinib (YN968D1, CAS 811803-05-1); imatinib (Gleevec®); ponatinib (AP24534, CAS 943319-70-8); tivozanib (AV951, CAS 475108-18-0); regorafenib (BAY73-4506, CAS 755037-03-7); vatalanib dihydrochloride (PTK787, CAS 212141-51-0); brivanib (BMS-540215, CAS 649735-46-6); vandetanib (Caprelsa® or AZD6474); motesaniba diphosphate (AMG706, CAS 857876-30-3, N-(2,3-dihydro-3,3-dimethyl-1H-indol-6-yl)-2-[(4-pyridinylmethyl)amino]3-pyridinecarboxamide, described in PCT Publication No. WO02/066470); dovitinib dilactic acid (TKI258, CAS 852433-84-2); linfanib (ABT869, CAS 796967-16-3); cabozantinib (XL184, CAS 849217-68-1); lestaurtinib (CAS 111358-88-4); N-[5-[[[5-(1,1-dimethylethyl)-2-oxazolyl]methyl]thio]-2thiazolyl]-4-piperidinecarboxamide (BMS38703, CAS 345627-80-7); (3R,4R)-4-amino-1-((4-((3methoxyphenyl)amino)pyrrolo[2,1-N1,2,4]triazin-5-yl)methyl)piperidin-3-ol (BMS690514) ; N-(3,4dichloro-2-fluorophenyl)-6-methoxy-7-[[(3aα,5β,6aα)-octahydro-2-methylcyclopenta[c]pyrrol-5-yl]methoxy]4-quinazolinamine (XL647, CAS 781613-23-8); 4-methyl-3-[[1-methyl-6-(3-pyridinyl)-1H-pyrazolo[3,4d]pyrimidin-4-yl]amino]-N-[3-(trifluoromethyl)phenyl]benzamide (BHG712 , CAS 940310-85-0) and aflibercept (Eylea®).

Exemplary anti-VEGF antibodies include, but are not limited to, a monoclonal antibody that binds to the same epitope as the anti-VEGF monoclonal antibody A4.6.1 produced by ATCC hybridoma HB 10709; recombinant humanized anti-VEGF monoclonal antibody prepared according to Presta et al. (1997) Cancer Res. 57:4593-4599. In one embodiment, the anti-VEGF antibody is bevacizumab (BV), also known as rhuMAb VEGF or AVASTIN®. It contains mutant human IgG1 framework regions and antigen-binding complementarity-determining regions from the mouse anti-hVEGF monoclonal antibody A.4.6.1, which blocks the binding of human VEGF to its receptors.

Bevacizumab and other humanized anti-VEGF antibodies are further described in US Pat. No. 6,884,879 issued February 26, 2005. Additional antibodies include antibodies of the G6 or B20 series (e.g., G6-31, B20-4.1) as described in PCT Publication No. WO 2005/012359, PCT Publication No. WO 2005/044853, the contents of these patent applications are incorporated herein by reference in in full. For additional antibodies, see US patents No. 7060269, 6582959, 6703020, 6054297, WO98/45332, Wo96/30046, WO94/10202, EP06668688B1, Publication of Patent Applications of US and Popkov et al., Journal of Immunological Methods 288: 149-164 (2004). Other antibodies include antibodies that bind to a functional epitope on human VEGF containing residues F17, M18, D19, Y21, Y25, Q89, 191, K101, E103 and C104 or alternatively containing residues F17, Y21, Q22, Y25, D63, 183 and Q89.

In some embodiments, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule described herein, is used alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PD antibody molecule). -L1), in combination with a PI3K inhibitor. In one embodiment, the PI3K inhibitor is an inhibitor of the delta and gamma isoforms of PI3K. Exemplary PI3K inhibitors that can be used in combination are described, for example, in WO 2010/036380, WO 2010/006086, WO 09/114870, WO 05/113556: GSK 2126458, GDC-0980, GDC-0941, Sanofi XL147, XL756 , XL147, PF-46915032, BKM 120, CAL-101, CAL 263, SF1126, PX-886, and a dual PI3K inhibitor (eg, Novartis BEZ235).

- 64 040365

In some embodiments, the anti-TIM-3 antibody molecules described herein are used, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PD-L1 antibody molecule), in combination with an inhibitor mTOR, e.g., one or more mTOR inhibitors selected from one or more of rapamycin, temsirolimus (TORISEL®), AZD8055, BEZ235, BGT226, XL765, PF-4691502, GDC0980, SF1126, OSI027, GSK1059615, KU-0063794, WYE- 354, palomide 529 (P529), PF-04691502 or PKI-587, ridaforolimus (formerly known as deferolimus, (1R,2R,4S)-4-[(2R)-2-[{1R,9S,12S,15R, 16E,18R,19R,21R, 23S,24E,26E,28Z,30S,32S,35R)-1,18-dihydroxu-19,30-dimethoxu-15,17,21,23,29,35-hexαmetul2,3 ,10,14,20-pentaoxo-11,36-dioxa-4-azatricyclo[30, 3, 1, 0 ^4,9 ]hexatriaconta-16, 24,26,28-tetraen-12yl]propyl]-2-methoxycyclohexyldimethylphosphinate , also known as AP23573 and MK8669 and described in PCT Publication No. WO03/064383); everolimus (Afinitor® or RAD001); rapamycin (AY22989, Sirolimus®); simapimoda (CAS 164301-51-3); emsirolimus, (5-{2,4-bis[(3S)-3-methylmorpholin-4yl]pyrido[2,3-d]pyrimidin-7-yl}-2-methoxyphenyl)methanol (AZD8055); 2-amino-8-[trans-4-(2-hydroxyethoxy)cyclohexyl]-6-(6-methoxy-3-pyridinyl)-4-methylpyrido[2,3-d]pyrimidin-7(8H)-one ( PF04691502, CAS 1013101-36-4); and N ² -[1,4-dioxo-4-[[4-(4-oxo-8-phenyl-4H-1-benzopyran-2-yl)morpholinium-4-yl]methoxy]butyl]-E-arginylglycyl -E-a-aspartyl-E-serine, internal salt (SF1126, CAS 936487-67-1) and XL765.

In some embodiments, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule described herein, is used alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PD antibody molecule). -L1), in combination with a BRAF inhibitor such as GSK211843 6, RG7204, PLX4032, GDC-0879, PLX4720 and sorafenib tosylate (Bay 43-9006).

In some embodiments, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule described herein, is used alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PD antibody molecule). -L1), in combination with a MEK inhibitor. In some embodiments, a combination of an anti-TIM-3 antibody and a MEK inhibitor is used to treat a cancer (eg, the cancer described herein). In some embodiments, the cancer being treated with the combination is selected from melanoma, colorectal cancer, non-small cell lung cancer, ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, hematologic malignancy, or renal cell carcinoma. In certain embodiments, the cancer comprises a BRAF mutation (eg, BRAF V600E mutation), wild-type BRAF, wild-type KRAS, or an activating KRAS mutation. A malignant tumor may be at an early, intermediate or late stage. Any MEK inhibitor can be used in combination, including but not limited to ARRY-142886, G02442104 (also known as GSK1120212), RDEA436, RDEA119/BAY 869766, AS703026, G00039805 (also known as AZD-6244 or selumetinib), BIX 02188 , BIX 02189, CI-1040 (PD184352), PD0325901, PD98059, U0126, GDC-0973 (Methanone, [3,4-difluoro-2-[(2-fluoro-4iodophenyl)amino]phenyl][3-hydroxy-3 -(25)-2-piperidinyl-1-azetidinyl]-), G-38963, G02443714 (also known as AS703206) or a pharmaceutically acceptable salt or solvate thereof. Additional examples of MEK inhibitors are described in WO 2013/019906, WO 03/077914, WO 2005/121142, WO 2007/04415, WO 2008/024725 and WO 2009/085983, the contents of which are incorporated herein by reference.

In some embodiments, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule described herein, is used alone or in combination with another immunomodulator (eg, an anti-LAG-3, anti-PD-1, or anti-PD- L1), in combination with a JAK2 inhibitor such as CEP-701, INCB18424, CP-690550 (tasocitinib).

In some embodiments, the pharmaceutical composition described herein is used, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-L1, or anti-PD-1 antibody molecule), in combination with paclitaxel or an agent based on paclitaxel, eg TAXOL®, protein-bound paclitaxel (eg ABRAXANE®). Exemplary paclitaxel-based agents include, but are not limited to, nanoparticulate albumin-bound paclitaxel (ABRAXANE, marketed by Abraxis Bioscience), docosahexaenoic acid-bound paclitaxel (DHA-paclitaxel, taxoprexin, marketed by Protarga), polyglutamate-bound paclitaxel (PG-paclitaxel, paclitaxel polyglumex, CT-2103, XYOTAX, marketed by Cell Therapeutic), tumor-activated prodrug (TAP), ANG105 (Angiopep-2 linked to three molecules of paclitaxel, marketed by ImmunoGen), paclitaxel-EC -1 (paclitaxel linked to erbB2 recognition peptide EC1; see Li et al., Biopolymers (2007) 87:225-230), and glucose-conjugated paclitaxel (e.g., 2'-paclitaxel methyl-2-glucopyranosyl succinate, see Liu et al., Bioorganic & Medicinal Chemistry Letters (2007) 17:617-620).

- 65 040365

Radiation therapy can be delivered by one of several methods or a combination of methods, including, but not limited to, external beam therapy, internal radiation therapy, implant-assisted radiation, stereotactic radiosurgery, systemic radiation therapy, radiation therapy, and permanent or temporary interstitial brachytherapy. The term brachytherapy refers to radiation therapy delivered by spatially confined radioactive material placed in or near a tumor or other area of tissue proliferative disease. The term is intended to include, but is not limited to, exposure to radioactive isotopes (e.g., At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P -32 and radioactive isotopes Lu). Suitable radiation sources for use as the cell treatment agent of the present invention include both solids and liquids. As a non-limiting example, a radiation source can be a radionuclide such as I-125, I-131, Yb-169, Ir-192 as a solid source, I-125 as a solid source, or other radionuclides that emit photons, beta particles, gamma radiation and other therapeutic beams. The radioactive material may also be a liquid made from any solution of the radionuclide(s), such as a solution of I-125 or I-131, or the radioactive liquid may be prepared using a slurry of a suitable liquid containing fine particles of solid radionuclides such as Au-198, Y-90. Moreover, the radionuclide(s) may be in the form of a gel or radioactive microspheres.

Anti-TIM-3 antibody molecules, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PD-L1 antibody molecule), may be administered in combination with one or more of the existing cancer treatments, including but not limited to: surgery; radiation therapy (eg, external beam therapy, which involves three-dimensional conformal radiation therapy, where the irradiation field is a calculated local irradiation (eg, irradiation directed to a preselected target or organ) or focused irradiation). Focused radiation may be selected from the group consisting of stereotactic radiosurgery, fractionated stereotactic radiosurgery, and intensity modulated radiation therapy. Focused irradiation may have a radiation source selected from the group consisting of a particle beam (proton), cobalt-60 (photon) and a linear accelerator (X-ray), for example, as described in WO 2012/177624.

In certain embodiments, an anti-TIM-3 antibody molecule, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PD-L1 antibody molecule), is administered in combination with an antibody against immunoglobulin-like killer cell receptors. cells (also referred to herein as an anti-KIR antibody), a general anti-KIR antibody or an anti-NKG2D antibody, and an anti-MICA antibody. In certain embodiments, a combination of an anti-TIM-3 antibody molecule and an anti-KIR antibody, a generic anti-KIR antibody, or an anti-NKG2D antibody described herein is used to treat a cancer, e.g., a cancer as described herein (e.g., a solid tumor e.g., an advanced solid tumor).

In one embodiment, an anti-TIM-3 antibody molecule, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PDL1 antibody molecule), is administered in combination with a cellular immunotherapy (e.g., provenge (e.g., sipuleucel)), and optionally in combination with cyclophosphamide. In certain embodiments, a combination of an anti-TIM-3 antibody molecule, provenge, and/or cyclophosphamide is used to treat a cancer, eg, a cancer as described herein (eg, prostate cancer, eg, advanced prostate cancer).

In another embodiment, an anti-TIM-3 antibody molecule, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PDL1 antibody molecule), is administered in combination with a vaccine, e.g. dendritic cells (DC-RCC). In certain embodiments, the combination of an anti-TIM-3 antibody molecule and a DC-RCC vaccine is used to treat a cancer, e.g., a cancer as described herein (e.g., liver carcinoma, e.g., metastatic renal cell carcinoma (RCC) or clear cell renal cell carcinoma ( CCRCC)).

In another embodiment, an anti-TIM-3 antibody molecule, alone or in combination with another immunomodulator (eg, an anti-LAG-3, anti-PD-1, or anti-PDL1 antibody molecule), is administered in combination with chemotherapy and/or immunotherapy. For example, an anti-TIM-3 antibody molecule can be used to treat myeloma, alone or in combination with one or more of: chemotherapy or other anti-cancer agents (e.g., thalidomide analogs, e.g., lenalidomide), anti-PD-1 antibodies treated with a tumor dendritic cell antigen, fusion (eg, electrofusion) of tumor cells and dendritic cells, or vaccination with an immunoglobulin idiotype produced by malignant plasma cells. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with an anti-PD-1 antibody for the treatment of myeloma, eg, multiple myeloma.

- 66 040365

In one embodiment, an anti-TIM-3 antibody molecule, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PDL1 antibody molecule), is used in combination with chemotherapy to treat lung cancer, e.g., non-small cell cancer. lung. In one embodiment, an anti-TIM-3 antibody molecule is used with platinum doublet therapy for the treatment of lung cancer.

In another embodiment, an anti-TIM-3 antibody molecule, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PDL1 antibody molecule), is used to treat kidney cancer, such as renal cell carcinoma (RCC) ( for example, clear cell renal cell carcinoma (CCRCC) or metastatic RCC The anti-TIM-3 antibody molecule can be administered in combination with one or more of: an immune-based strategy (eg, interleukin-2 or interferon-α), a targeted agent (eg, an inhibitor VEGF, such as an anti-VEGF monoclonal antibody; an inhibitor of VEGF tyrosine kinase, such as sunitinib, sorafenib, axitinib, and pazopanib; an RNAi-based inhibitor) or an inhibitor of a downstream VEGF signaling mediator, e.g., a mammalian target of rapamycin (mTOR), e.g. , everolimus and temsirolimus).

An example of suitable therapeutic agents for use in combination with the anti-TIM-3 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PD-L1 antibody molecule), for the treatment of pancreatic cancer includes, but is not limited to, a chemotherapeutic agent, such as paclitaxel or a paclitaxel-based agent (eg, a paclitaxel formulation such as TAXOL, an albumin-stabilized formulation of paclitaxel in the form of nanoparticles (eg, ABRAXANE) or a paclitaxel liposomal formulation); gemcitabine (eg, gemcitabine alone or in combination with AChP107-11); other chemotherapeutic agents such as oxaliplatin, 5-fluorouracil, capecitabine, rubitecan, epirubicin hydrochloride, NC-6004, cisplatin, docetaxel (eg TAXOTERE), mitomycin C, ifosfamide; interferon; tyrosine kinase inhibitor (eg, EGFR inhibitor (eg, erlotinib, panitumumab, cetuximab, nimotuzumab); HER2/neu receptor inhibitor (eg, trastuzumab); dual kinase inhibitor (eg, bosutinib, saracatinib, lapatinib, vandetanib); multikinase inhibitor (eg, sorafenib, sunitinib, XL184, pazopanib); VEGF inhibitor (eg, bevacizumab, AV-951, brivanib); radioimmunotherapy (eg, XR303); cancer vaccine (eg, GVAX, survivin peptide); COX-2 inhibitor (eg, celecoxib); IGF-1 receptor inhibitor (eg, AMG 479, MK-0646); mTOR inhibitor (eg, everolimus, temsirolimus); IL-6 inhibitor (eg, CNTO 328); cyclin-dependent kinase inhibitor (eg, P276- 00, UCN-01); targeted altered energy metabolism (AEMD) compound (eg, CPI-613); HDAC inhibitor (eg, vorinostat); TRAIL receptor 2 (TR-2) agonist (eg, conatumumab); MEK inhibitor ( e.g. AS703026, selumetinib, GSK1120212); double inhibition itor kinase Raf/MEK (eg RO5126766); a Notch signaling inhibitor (eg, MK0752); fusion protein monoclonal antibody-antibody (for example, L19IL2); curcumin; an HSP90 inhibitor (eg tanespimycin, STA-9090); rIL-2; denileukin diftitox; a topoisomerase 1 inhibitor (eg, irinotecan, PEP02); a statin (eg simvastatin); a factor VIIa inhibitor (eg, PCI-27483); an AKT inhibitor (eg RX-0201); hypoxia-activated prodrug (eg, TH-302); metformin hydrochloride, gamma secretase inhibitor (eg RO4929097); a ribonucleotide reductase inhibitor (eg, 3-AP); immunotoxin (eg HuC242-DM4); a PARP inhibitor (eg KU0059436, veliparib); a CTLA-4 inhibitor (eg, CP-675,206, ipilimumab); AdV-tk therapy; a proteasome inhibitor (eg, bortezomib (Velcade), NPI-0052); thiazolidinedione (eg pioglitazone); NPC-1C; Aurora kinase inhibitor (eg R763/AS703569), CTGF inhibitor (eg FG-3019); siG12D LODER; and radiation therapy (eg, tomotherapy, seterotactic radiation, proton therapy), surgery, and a combination thereof. In certain embodiments, a combination of paclitaxel or a paclitaxel and gemcitabine agent can be used with the anti-TIM-3 antibody molecules described herein.

An example of suitable therapeutic agents for use in combination with anti-TIM-3 antibody molecules, alone or in combination with another immunomodulator (e.g. anti-LAG-3, anti-PD-1 or anti-PD-L1 antibody molecule), for the treatment of small cell lung cancer includes but not limited to, a chemotherapeutic agent (eg, etoposide, carboplatin, cisplatin, oxaliplatin, irinotecan, topotecan, gemcitabine, SN-38 liposomal, bendamustine, temozolomide, belotecan, NK012, FR901228, flavopiridol); tyrosine kinase inhibitor (eg, EGFR inhibitor (eg, erlotinib, gefitinib, cetuximab, panitumumab); multikinase inhibitor (eg, sorafenib, sunitinib); VEGF inhibitor (eg, bevacizumab, vandetanib); cancer vaccine (eg, GVAX); inhibitor Bcl-2 (eg, oblimersen sodium, ABT-263), proteasome inhibitor (eg, bortezomib (Velcade), NPI-0052), paclitaxel or paclitaxel-based agent, docetaxel, IGF-1 receptor inhibitor (eg, AMG 479); HGF/SF inhibitor (eg, AMG 102, MK-0646); chloroquine; Aurora kinase inhibitor (eg, MLN8237); radioimmunotherapy (eg, TF2); HSP90 inhibitor (eg, tanespimycin, STA-9090); mTOR inhibitor (for

- 67 040365 example, everolimus); a bispecific anti-eEp-CAM/CD3 antibody (eg, MT110); a CK-2 inhibitor (eg CX-4945); an HDAC inhibitor (eg, belinostat); an SMO antagonist (eg BMS 833923); cancer peptide vaccine; and radiation therapy (eg, intensity-modulated radiation therapy (IMRT), hypofractionated radiation therapy, hypoxia-guided radiation therapy), surgery, and combinations thereof.

An example of suitable therapeutic agents for use in combination with anti-TIM-3 antibody molecules, alone or in combination with another immunomodulator (e.g. anti-LAG-3, anti-PD-1 or anti-PD-L1 antibody molecule), for the treatment of non-small cell lung cancer includes but not limited to, a chemotherapeutic agent (e.g., vinorelbine, cisplatin, docetaxel, pemetrexed disodium, etoposide, gemcitabine, carboplatin, liposomal SN-38, TLK286, temozolomide, topotecan, pemetrexed disodium, azacitidine, irinotecan, tegafur-gimeracil-oteracil potassium , sapacitabine); tyrosine kinase inhibitor (eg, EGFR inhibitor (eg, erlotinib, gefitinib, cetuximab, panitumumab, necitumumab, PF-00299804, nimotuzumab, RO5083945)), MET inhibitor (eg, PF-02341066, ARQ 197), PI3K kinase inhibitor (eg, XL147, GDC-0941), dual Raf/MEK kinase inhibitor (eg, RO5126766), dual PI3K/mTOR kinase inhibitor (eg, XL765), SRC inhibitor (eg, dasatinib), dual inhibitor (eg, BIBW 2992, GSK1363089, ZD6474 , AZD0530, AG-013736, lapatinib, MEHD7945A, linifanib), multikinase inhibitor (eg, sorafenib, sunitinib, pazopanib, AMG 706, XL184, MGCD265, BMS-690514, R935788), VEGF inhibitor (eg, endostar, endostatin, bevacizumab, cediranib, BIBF 1120, axitinib, tivozanib, AZD2171), cancer vaccine (eg, BLP25 liposomal vaccine, GVAX, recombinant DNA and adenovirus expressing L523S protein), Bcl-2 inhibitor (eg, oblimersen sodium), proteasome inhibitor (eg , bortezomib, carcilzomib, NPI-0052, MLN9708), paclitaxel or paclitaxel-based agent, docetaxel, IGF-1 receptor inhibitor (eg, cixutumumab, MK-0646, OSI 906, CP-751,871, BIIB022), hydroxychloroquine, HSP90 inhibitor (eg, tanespimycin, STA -9090, AUY922, XL888), mTOR inhibitor (eg, everolimus, temsirolimus, ridaphorolimus), anti-Ep-CAM-/CD3 bispecific antibody (eg, MT110), CK-2 inhibitor (eg, CX-4945), HDAC inhibitor ( e.g. MS 275, LBH589, vorinostat, valproic acid, FR901228), DHFR inhibitor (e.g. pralatrexate), retinoid (e.g. bexarotene, tretinoin), antibody drug conjugate (e.g. SGN-15), bisphosphonate (e.g. zoledronic acid), cancer vaccine (eg, belagenpumatucel-L), low molecular weight heparin (LMWH) (eg, tinzaparin, enoxaparin), GSK1572932A, melatonin, talactoferrin, dimesnu, topoisomerase inhibitor (eg, amrubicin, etoposide, carenitecin), nelfinavir, cilengitide, an ErbB inhibitor 3 (eg, MM-121, U3-1287), survivin inhibitor (eg, YM155, LY2181308), eribulin mesylate, COX-2 inhibitor (eg, celecoxib), pegfilgrastim, Polo-like kinase 1 inhibitor (eg, BI 6727) , TRAIL receptor 2 (TR-2) agonist (e.g. CS-1008), CNGRC peptide conjugate (SEQ ID NO: 225)-TNF-alpha, dichloroacetate (DCA), HGF inhibitor (e.g. SCH 900105), SAR240550, agonist PPAR-gamma (eg, CS-7017), gamma secretase inhibitor (eg, RO4929097), epigenetic therapy (eg, 5-azacytidine), nitroglycerin, MEK inhibitor (eg, AZD6244), cyclin-dependent kinase inhibitor (eg, UCN -01), cholesterol-Fusl, antitubulin (eg, E7389), farnesyl-OH transferase inhibitor (eg, lonafarnib), immunotoxin (eg, BB-10901, SSI (dsFv) PE38), fondaparinux, vasoconstrictor ( e.g. AVE8062), PD-L1 inhibitor (e.g. MDX-1105, MDX-1106), beta-glucan, NGRhTNF, EMD 521873, MEK inhibitor (e.g. GSK1120212), epothilone analog (on ixabepilone), kinesin spindle inhibitor (eg 4SC-205), telomere targeting agent (eg KML-001), P70 cascade inhibitor (eg LY2584702), AKT inhibitor (eg MK-2206), angiogenesis inhibitor (eg eg, lenalidomide), a Notch signaling inhibitor (eg, OMP21M18), radiation therapy, surgery, and combinations thereof.

An example of suitable therapeutic agents for use in combination with anti-TIM-3 antibody molecules, alone or in combination with another immunomodulator (e.g. anti-LAG-3, anti-PD-1 or anti-PD-L1 antibody molecule), for the treatment of ovarian cancer includes, but not limited to, a chemotherapeutic agent (e.g., paclitaxel or paclitaxel-based agent; docetaxel; carboplatin; gemcitabine; doxorubicin; topotecan; cisplatin; irinotecan, TLK286, ifosfamide, olaparib, oxaliplatin, melphalan, pemetrexed disodium, SJG-136, cyclophosphamide, etoposide, decitabine); ghrelin antagonist (eg, AEZS-130), immunotherapy (eg, APC8024, oregovomab, ORT-821), tyrosine kinase inhibitor (eg, EGFR inhibitor (eg, erlotinib), dual inhibitor (eg, E7080), multikinase inhibitor (eg, AZD0530 , JI-101, sorafenib, sunitinib, pazopanib), ON 01910.Na), VEGF inhibitor (eg, bevacizumab, BIBF 1120, cediranib, AZD2171), PDGFR inhibitor (eg, IMC-3G3), paclitaxel, topoisomerase inhibitor (eg, carenitecin, irinotecan), HDAC inhibitor (eg, valproate, vorinostat), folate receptor inhibitor (eg, farletuzumab), angiopoietin inhibitor (eg, AMG 386), epothilone analogue (eg, ixabepilone), proteasome inhibitor (eg, carfilzomib), inhibitor IGF-1 receptor (eg OSI 906, AMG 479), PARP inhibitor (eg veliparib, AG014699, iniparib,

- 68 040365

MK-4827), Aurora kinase inhibitor (eg, MLN8237, ENMD-2076), angiogenesis inhibitor (eg, lenalidomide), DHFR inhibitor (eg, pralatrexate), radioimmunotherapy (eg, Hu3S193), statin (eg, lovastatin), topoisomerase 1 inhibitor (eg, NKTR-102), cancer vaccine (eg, p53 synthetic long peptide vaccine, autologous OS-DC vaccine), mTOR inhibitor (eg, temsirolimus, everolimus), BCR/ABL inhibitor ( eg, imatinib), ET-A receptor antagonist (eg, ZD4054), TRAIL receptor 2 (TR-2) agonist (eg, CS-1008), HGF/SF inhibitor (eg, AMG 102), EGEN-001, Polo inhibitor -like kinase 1 (eg, BI 6727), gamma secretase inhibitor (eg, RO4929097), Wee-1 inhibitor (eg, MK-1775), antitubulin agent (eg, vinorelbine, E7389), immunotoxin (eg, denileukin diftitox) , SB-485232, vascular disruptor (eg, AVE8062), integrin inhibitor (n eg EMD 525797), kinesin spindle inhibitor (eg 4SC-205), revlimid, HER2 inhibitor (eg MGAH22), ErrB3 inhibitor (eg MM-121), radiotherapy; and their combinations.

In one exemplary embodiment, an anti-TIM-3 antibody molecule, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PD-L1 antibody molecule) is used to treat myeloma, alone or in combination with one or several of: chemotherapy or other anti-cancer agents (eg, thalidomide analogs, eg lenalidomide), HSCT (Cook, R. (2008) J Manag Care Pharm. 14(7 Suppl):19-25), anti-TIM3 antibodies (Hallett, WHD et al. (2011) J of American Society for Blood and Marrow Transplantation 17(8):1133-145), tumor antigen-treated dendritic cells, fusion (eg, electrofusion) of tumor cells and dendritic cells, or vaccination with an idiotype immunoglobulin produced by malignant plasma cells (reviewed in Yi, Q. (2009) Cancer J. 15(6):50210).

In another embodiment, an anti-TIM-3 antibody molecule, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PDL1 antibody molecule), is used to treat kidney cancer, such as renal cell carcinoma (RCC) or metastatic RCC. The anti-TIM-3 antibody molecule can be administered in combination with one or more of: an immune-based strategy (eg, interleukin-2 or interferon-α), a targeted agent (eg, a VEGF inhibitor such as an anti-VEGF monoclonal antibody, such as bevacizumab ( Rini, B. I. et al (2010) J Clin Oncol 28(13):2137-2143)); a VEGF tyrosine kinase inhibitor such as sunitinib, sorafenib, axitinib, and pazopanib (reviewed in Pal. S.K. et al. (2014) Clin. Advances in Hematology & Oncology 12(2):90-99)); an RNAi inhibitor) or an inhibitor of a downstream mediator of VEGF signaling, e.g., a mammalian target of rapamycin (mTOR), e.g., everolimus and temsirolimus (Hudes, G. et al. (2007) N. Engl. J. Med. 356(22 ):2271-2281, Motzer, R. J. et al (2008) Lancet 372:449-456).

An example of suitable therapeutic agents for use in combination with the anti-TIM-3 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PD-L1 antibody molecule), for treatment of chronic myelogenous leukemia (AML) according to the invention includes, but is not limited to, a chemotherapeutic agent (e.g., cytarabine, hydroxyurea, clofarabine, melphalan, thiotepa, fludarabine, busulfan, etoposide, cordycepin, pentostatin, capecitabine, azacitidine, cyclophosphamide, cladribine, topotecan ), tyrosine kinase inhibitor (eg, BCR/ABL inhibitor (eg, imatinib, nilotinib), ON 01910.Na, dual inhibitor (eg, dasatinib, bosutinib), multikinase inhibitor (eg, DCC-2036, ponatinib, sorafenib, sunitinib, RGB -286638)), interferon-alpha, steroids, apoptotic agent (eg, omacetaxin mepesuccinate), immunotherapy (eg, allogeneic CD4+ Th1-like memory T cells /microparticle-bound anti-CD3/anti-CD28 antibody, autologous cytokine-inducible killer cells (CIK), AHN-12), CD52-targeting agent (e.g., alemtuzumab), HSP90 inhibitor (e.g., tanespimycin, STA9090, AUY922, XL888), inhibitor mTOR (eg, everolimus), SMO antagonist (eg, BMS 833923), ribonucleotide reductase inhibitor (eg, 3-AP), JAK-2 inhibitor (eg, INCB018424), hydroxychloroquine, retinoid (eg, fenretinide), cyclin-dependent kinase inhibitor (e.g. UCN-01), HDAC inhibitor (e.g. belinostat, vorinostat, JNJ-26481585), PARP inhibitor (e.g. veliparib), MDM2 antagonist (e.g. RO5045337), Aurora B kinase inhibitor (e.g. TAK-901), radioimmunotherapy (eg, actinium 225-labeled anti-CD33 antibody HuM195), Hedgehog inhibitor (eg, PF-04449913), STAT3 inhibitor (eg, OPB-31121), KB004, cancer vaccine (eg, AG858), bone marrow transplant, transplant stem cells, radiation therapy, and combinations thereof.

An example of suitable therapeutic agents for use in combination with anti-TIM-3 antibody molecules, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD-1, or anti-PD-L1 antibody molecule), for the treatment of chronic lymphocytic leukemia ( CLL) includes, but is not limited to, a chemotherapeutic agent (e.g., fludarabine, cyclophosphamide, doxorubicin, vincristine, chlorambucil, bendamustine, chlorambucil, busulfan, heme

- 69 040365 cytabin, melphalan, pentostatin, mitoxantrone, 5-azacytidine, pemetrexed disodium), tyrosine kinase inhibitor (eg EGFR inhibitor (eg erlotinib), BTK inhibitor (eg PCI-32765), multikinase inhibitor (eg MGCD265, RGB -286638), CD-20 targeting agent (eg, rituximab, ofatumumab, RO5072759, LFB-R603), CD52 targeting agent (eg, alemtuzumab), prednisolone, darbepoetin-alpha, lenalidomide, Bcl-2 inhibitor (eg, ABT -263), immunotherapy (eg, allogeneic CD4+ Th1-like memory T cells/microparticle-associated anti-CD3/anti-CD28 antibody, autologous cytokine-inducible killer cells (CIK)), HDAC inhibitor (eg, vorinostat, valproic acid, LBH589, JNJ-26481585, AR-42), XIAP inhibitor (eg, AEG35156), CD-74 targeting agent (eg, milatuzumab), mTOR inhibitor (eg, everolimus), AT-101, immunotoxin (eg, CAT-8015, anti -Tac(Fv)-PE38 (LMB2)), CD37 targeting agent (e.g. p, TRU-016), radioimmunotherapy (eg, 131tositumomab), hydroxychloroquine, perifosine, SRC inhibitor (eg, dasatinib), thalidomide, PI3K delta inhibitor (eg, CAL-101), retinoid (eg, fenretinide), MDM2 antagonist ( e.g. RO5045337), plerixaphor, Aurora kinase inhibitor (e.g. MLN8237, TAK-901), proteasome inhibitor (e.g. bortezomib), CD-19 targeting agent (e.g. MEDI-551, MOR208), MEK inhibitor (e.g., ABT-348), JAK-2 inhibitor (eg, INCB018424), hypoxia-activated prodrug (eg, TH-302), paclitaxel or paclitaxel-based agent, HSP90 inhibitor, AKT inhibitor (eg, MK2206), HMG-CoA inhibitor (eg. , simvastatin), GNKG186, radiation therapy, bone marrow transplantation, stem cell transplantation, and a combination thereof.

An example of suitable therapeutic agents for use in combination with the anti-TIM-3 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PD-L1 antibody molecule), for treatment of acute lymphocytic leukemia (ALL) includes, but is not limited to, a chemotherapeutic agent (e.g., prednisolone, dexamethasone, vincristine, asparaginase, daunorubicin, cyclophosphamide, cytarabine, etoposide, thioguanine, mercaptopurine, clofarabine, liposomal annamycin, busulfan, etoposide, capecitabine, decitabine, azacitidine, topotecan, temozolomide), tyrosine kinase inhibitor (eg, BCR/ABL inhibitor (eg, imatinib, nilotinib), ON 01910.Na, multikinase inhibitor (eg, sorafenib)) CD-20 targeting agent (eg, rituximab ), CD52 targeting agent (eg, alemtuzumab), HSP90 inhibitor (eg, STA-9090), mTOR inhibitor (eg, everolimus, rapamycin), JAK-2 inhibitor (n eg, INCB018424), HER2/neu receptor inhibitor (eg, trastuzumab), proteasome inhibitor (eg, bortezomib), methotrexate, asparaginase, CD-22 targeting agent (eg, epratuzumab, inotuzumab), immunotherapy (eg, autologous cytokine-inducible killer cells (CIK), AHN-12), bilanatumomab, cyclin-dependent kinase inhibitor (eg UCN-01), CD45 targeting agent (eg BC8), MDM2 antagonist (eg RO5045337), immunotoxin (eg CAT-8015 , DT2219ARL), HDAC inhibitor (eg, JNJ-26481585), JVRS-100, paclitaxel or paclitaxel-based agent, STAT3 inhibitor (eg, OPB-31121), PARP inhibitor (eg, veliparib), EZN-2285, radiotherapy, steroid, bone marrow transplant, stem cell transplant, or a combination thereof.

An example of suitable therapeutic agents for use in combination with the anti-TIM-3 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PD-L1 antibody molecule), for treatment of acute myeloid leukemia (AML) includes, but is not limited to, a chemotherapeutic agent (eg, cytarabine, daunorubicin, idarubicin, clofarabine, decitabine, vosaroxin, azacitidine, clofarabine, ribavirin, CPX-351, treosulfan, elacytarabine, azacitidine), tyrosine kinase inhibitor (eg, BCR/ABL inhibitor (eg, imatinib, nilotinib), ON 01910.Na, multikinase inhibitor (eg, midostaurin, SU 11248, quisartinib, sorafinib)), immunotoxin (eg, gemtuzumab ozagomycin), DT388IL3 fusion protein, HDAC inhibitor (eg, vorinostat, LBH589), plerixafor, mTOR inhibitor (eg, everolimus), SRC inhibitor (eg, dasatinib), HSP90 inhibitor (eg, STA-9090), retinoid (eg, bexar oten, Aurora kinase inhibitor (eg, BI 811283), JAK-2 inhibitor (eg, INCB018424), Polo-like kinase inhibitor (eg, BI 6727), cenersen, CD45 targeting agent (eg, BC8), cyclin- dependent kinase (eg, UCN-01), MDM2 antagonist (eg, RO5045337), mTOR inhibitor (eg, everolimus), LY573636-sodium, ZRx-101, MLN4924, lenalidomide, immunotherapy (eg, AHN-12), histamine dihydrochloride, radiation therapy, bone marrow transplantation, stem cell transplantation, and a combination thereof.

An example of suitable therapeutic agents for use in combination with the anti-TIM-3 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1, or anti-PD-L1 antibody molecule), for treatment of multiple myeloma (MM) includes, but is not limited to, a chemotherapeutic agent (e.g., melphalan, amifostine, cyclophosphamide, doxorubicin, clofarabine, bendamustine, fludarabine, adriamycin, SyB L-0501), thalidomide, lenalidomide, dexamethasone, prednisone, pomalidomide, proteasome inhibitor (eg, bortezomib, carfilzomib, MLN9708), cancer vaccine

- 70 040365 (eg GVAX), CD-40 targeting agent (eg SGN-40, CHIR-12.12), perifosine, zoledronic acid, immunotherapy (eg Mage-Az, NY-ESO-1, HuMax-CD38) , HDAC inhibitor (eg, vorinostat, LBH589, AR-42), aplidine, cyclin-dependent kinase inhibitor (eg, PD0332991, dinaciclib), arsenic trioxide, CB3304, HSP90 inhibitor (eg, KW-2478), tyrosine kinase inhibitor (eg, EGFR inhibitor (eg, cetuximab), multikinase inhibitor (eg, AT9283)), VEGF inhibitor (eg, bevacizumab), plerixaphor, MEK inhibitor (eg, AZD6244), IPH2101, atorvastatin, immunotoxin (eg, BB-10901), NPI- 0052, radioimmunotherapy (eg, yttrium Y90 ibritutomab tiuxetan), STAT3 inhibitor (eg, OPB-31121), MLN4924, Aurora kinase inhibitor (eg, ENMD-2076), IMGN901, ACE-041, CK-2 inhibitor (eg, CX-4945), radiation therapy, bone marrow transplantation, stem cell transplantation, and combinations thereof.

An example of suitable therapeutic agents for use in combination with anti-TIM-3 antibody molecules, alone or in combination with another immunomodulator (e.g. anti-LAG-3, anti-PD-1 or anti-PD-L1 antibody molecule), for the treatment of prostate cancer includes but not limited to, chemotherapy (eg, docetaxel, carboplatin, fludarabine), abiraterone, hormonal t (eg, flutamide, bicalutamide, nilutamide, cyproterone acetate, ketoconazole, aminoglutethimide, abarelix, degarelix, leuprolide, goserelin, triptorelin, buserelin) , tyrosine kinase inhibitor (eg, dual kinase inhibitor (eg, lapatanib), multikinase inhibitor (eg, sorafenib, sunitinib)), VEGF inhibitor (eg, bevacizumab), TAK-700, cancer vaccine (eg, BRX-101, PEP223), lenalidomide, TOK-001, IGF-1 receptor inhibitor (eg, cixutumumab), TRC105, Aurora A kinase inhibitor (eg, MLN8237), proteasome inhibitor (eg, bortezomib), OGX-011, radioimmuno notherapy (eg, HuJ591-GS), HDAC inhibitor (eg, valproic acid, SB939, LBH589), hydroxychloroquine, mTOR inhibitor (eg, everolimus), dovininib lactate, diindolylmethane, efavirenz, OGX-427, genistein, IMC-3G3, bafetinib , CP675.206, radiation therapy, surgery, or a combination thereof.

An example of suitable therapeutic agents for use in combination with anti-TIM-3 antibody molecules alone or in combination with another immunomodulator (e.g., anti-LAG-3 antibody molecule, anti-PD-L1 antibody, or anti-PD-1 antibody molecule) for the treatment of HNSCC includes, but not limited to, one or both of compound A8 as described herein (or the compound described in PCT Publication No. WO 2010/029082) and cetuximab (eg Erbitux, marketed by BMS). In some embodiments, the therapeutic agent (eg, compound A8 or an A8 related compound) is a PI3K modulator, such as a PI3K inhibitor. In some embodiments, the therapeutic agent (eg, cetuximab) modulates, eg, inhibits, EGFR. In some embodiments, the cancer has or is identified as having elevated levels or activity of PI3K or EGFR compared to a control cell or reference value.

Example of suitable therapeutic agents for use in combination with anti-TIM-3 antibody molecules alone or in combination with another immunomodulator (eg, anti-LAG-3, anti-PD-L1, or anti-PD-1 antibody molecule) for the treatment of gastric cancer, e.g. gastric cancer with high MSI and/or EBV+ includes, but is not limited to, Compound A8 as described herein (or the compound described in PCT Publication No. WO 2010/029082). In some embodiments, the therapeutic agent (eg, Compound A8 or an A8 related compound) is a PI3K modulator, such as a PI3K inhibitor. In some embodiments, the cancer has or is identified as having elevated PI3K levels or activity compared to a control cell or reference value.

Example of suitable therapeutic agents for use in combination with anti-TIM-3 antibody molecules, alone or in combination with another immunomodulator (e.g. anti-LAG-3, anti-PD-L1 or anti-PD-1 antibody molecule), for the treatment of gastric cancer, e.g. gastric cancer with high MSI and/or inactivated RNF43 includes, but is not limited to, Compound A28 as described herein (or the compound described in PCT Publication No. WO 2010/101849). In some embodiments, the therapeutic agent (eg, compound A28 or an A28-related compound) is a modulator, such as an inhibitor, of porcupine. In some embodiments, the cancer has or is identified as having elevated levels or activity of porcupin compared to a control cell or reference value.

Example of suitable therapeutic agents for use in combination with anti-TIM-3 antibody molecules, alone or in combination with another immunomodulator (e.g. anti-LAG-3, anti-PD-L1 or anti-PD-1 antibody molecule) for the treatment of GI stromal tumor (GIST ) includes, but is not limited to, compound A16 as described herein (or the compound described in PCT Publication No. WO 1999/003854). In some embodiments, the therapeutic agent (eg, compound A16 or an A16-related compound) is a modulator, such as an inhibitor, of tyrosine kinase. In some embodiments, a malignant

- 71 040365 the tumor has, or is determined to have, elevated tyrosine kinase levels or activity compared to a control cell or reference value.

An example of suitable therapeutic agents for use in combination with anti-TIM-3 antibody molecules, alone or in combination with another immunomodulator (e.g. anti-LAG-3, anti-PD-L1 or anti-PD-1 antibody molecule), for the treatment of NSCLC, for example, squamous cell carcinoma or adenocarcinoma includes, but is not limited to, one or both of Compound A17 as described herein (or the compound described in US Pat. Nos. 7,767,675 and 8,420,645) and Compound A23 as described herein (or a compound described in PCT Publication No. WO 2003/077914). In some embodiments, a compound (eg, an A17 compound or an A17-related compound) modulates, eg, inhibits, c-MET. In some embodiments, a compound (eg, an A23 compound or an A23-related compound) modulates, eg, inhibits, Alk. In some embodiments, the cancer has or is determined to have elevated levels or activity of one or both of c-MET or Alk compared to a control cell or reference value. In some embodiments, the cancer has or is identified as having a mutation in EGFR.

An example of suitable therapeutic agents for use in combination with anti-TIM-3 antibody molecules alone or in combination with another immunomodulator (eg, anti-LAG-3, anti-PD-L1, or anti-PD-1 antibody molecule) for the treatment of melanoma (eg, NRAS melanoma ) includes, but is not limited to, one or both of compound A24 as described herein (or the compound described in US Pat. No. WO 2003/077914). In some embodiments, a compound (eg, an A24 compound or an A24-related compound) modulates, eg, inhibits, one or more of JAK and CDK4/6. In some embodiments, a compound (eg, an A34 compound or an A34-related compound) modulates, eg, inhibits, MEK. In some embodiments, the cancer has or is identified as having elevated levels or activity of one or more of JAK, CDK4/6, and MEK compared to a control cell or reference value.

An example of suitable therapeutic agents for use in combination with anti-TIM-3 antibody molecules, alone or in combination with another immunomodulator (e.g. anti-LAG-3, anti-PD-L1 or anti-PD-1 antibody molecule), for the treatment of melanoma (e.g., NRAS melanoma) includes, but is not limited to, one or both of Compound A29 as described herein (or compound described in PCT Publication No. WO 2011/025927) and Compound A34 as described herein (or compound described in PCT Publication No. WO 2003/077914). In some embodiments, a compound (eg, an A29 compound or an A29-related compound) modulates, eg, inhibits, BRAF. In some embodiments, a compound (eg, an A34 compound or an A34-related compound) modulates, eg, inhibits, MEK. In some embodiments, the cancer has or is identified as having elevated levels or activity of one or both of BRAF and MEK compared to a control cell or reference value.

An example of suitable therapeutic agents for use in combination with anti-TIM-3 antibody molecules alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD-L1, or anti-PD-1 antibody molecule) for the treatment of squamous NSCLC includes, but does not limited to them, the connection A5, as described in the present description (or the connection described in US patent No. 8552002). In some embodiments, a compound (eg, an A5 compound or an A5-related compound) modulates, eg, inhibits, FGFR. In some embodiments, the cancer has or is identified as having elevated FGFR levels or activity compared to a control cell or reference value.

An example of suitable therapeutic agents for use in combination with anti-TIM-3 antibody molecules alone or in combination with another immunomodulator (eg, anti-LAG-3, anti-PD-L1, or anti-PD-1 antibody molecule) for the treatment of colorectal cancer includes , but not limited to, one or both of compound A29 as described herein (or a compound according to PCT Publication No. WO 2011/025927) and cetuximab (eg Erbitux, marketed by BMS). In some embodiments, the therapeutic agent (eg, compound A29 or an A29-related compound) modulates, eg, inhibits, BRAF. In some embodiments, the therapeutic agent (eg, cetuximab) modulates, eg, inhibits EGFR. In some embodiments, the cancer has or is identified as having elevated levels or activity of BRAF or EGFR compared to a control cell or reference value.

The present invention also relates to a method of treating cancer with Compound A8, cetuximab and an anti-TIM-3 antibody molecule (optionally in combination with an anti-PD-1 antibody or an anti-LAG-3 antibody molecule). In some embodiments, the patient is first treated with compound A8 and cetuximab. This treatment continues for some

- 72 040365 amount of time, for example within a given amount of time, for example within about 1, 2, 4, 6, 8, 10 or 12 months. An anti-TIM-3 antibody molecule is then administered (optionally in combination with an anti-PD-1 antibody molecule or an anti-LAG-3 antibody molecule). The anti-TIM-3 antibody may optionally be administered in combination with cetuximab.

In some embodiments, the patient is first treated with all three of Compound A8, cetuximab, and an anti-TIM-3 antibody molecule (optionally in combination with an anti-PD-1 antibody or an anti-LAG-3 antibody molecule). This treatment is continued for an amount of time, such as a predetermined amount of time, such as approximately 6, 8, 10 or 12 months. Further, Compound A8 and/or cetuximab can be waived such that the maintenance phase involves treatment with an anti-TIM-3 antibody molecule (e.g., as monotherapy or in combination with an anti-PD-1 antibody or an anti-LAG-3 antibody molecule), but not compound A8 or cetuximab.

In other embodiments, the three compounds (compound A8, cetuximab, and an anti-TIM-3 antibody molecule, optionally in combination with an anti-PD-1 antibody or an anti-LAG-3 antibody molecule) are administered sequentially at the start of treatment. For example, Compound A8 and cetuximab may be administered first as described above. An anti-TIM-3 antibody molecule (optionally in combination with an anti-PD-1 antibody or an anti-LAG3 antibody molecule) is then added to the regimen. Further connection A8 and/or cetuximab can be canceled as described above.

Exemplary dosage regimens with three (or more) agents are as follows. The anti-TIM-3 antibody molecule can be administered, for example, at a dose of about 1 to 40 mg/kg, such as 1 to 30 mg/kg, such as about 5 to 25 mg/kg, about 10 to 20 mg/kg, about 1 to 5 mg/kg, or about 3 mg/kg. In some embodiments, Compound A8 is administered at a dose of about 200-300, 300-400, or 200-400 mg. In some embodiments, cetuximab is administered at an initial dose of 400 mg/m ² as an intravenous infusion over 120 minutes, followed by a 250 mg/m ² infusion every week for 60 minutes. In embodiments, one or more of Compounds A8, cetuximab, and an anti-TIM-3 antibody molecule is administered at a dose that is lower than the dose at which the agent would normally be administered as monotherapy, e.g., by about 0-10%, 10- 20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80% or 80-90% lower than the dose at which the agent is usually administered as monotherapy . In embodiments, one or more of compound A8, cetuximab, and an anti-TIM-3 antibody molecule is administered at a dose that is lower than the dose of this agent specified in this paragraph, for example, by about 0-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80% or 80-90% lower than the dose of this remedy indicated in this paragraph. In certain embodiments, the concentration of Compound A8 that is required to achieve inhibition, such as growth inhibition, is lower when Compound A8 is administered in combination with one or both of cetuximab and an anti-TIM-3 antibody molecule than when Compound A8 is administered alone. In certain embodiments, the concentration of cetuximab that is required to achieve inhibition, such as growth inhibition, is lower when cetuximab is administered in combination with one or both of Compound A8 and an anti-TIM-3 antibody molecule than when cetuximab is administered alone. In certain embodiments, the concentration of the anti-TIM-3 antibody molecule that is required to achieve inhibition, e.g., growth inhibition, is lower when the anti-TIM-3 antibody molecule is administered in combination with one or both of cetuximab and Compound A8 than when the antibody molecule against TIM-3 is administered individually.

In addition, within the scope of the present invention, a method for treating cancer with anti-TIM-3 antibody molecules alone or in combination with another immunomodulator (for example, an anti-LAG-3, anti-PD-L1, or anti-PD-1 antibody molecule) and a targeted anti-cancer agent is described. tumors, for example an agent that targets one or more proteins. In some embodiments, the anti-TIM-3 antibody molecule (and optionally other immunomodulator(s)) is administered first, followed by the targeted cancer agent. The length of time between administration of the anti-TIM-3 antibody molecule and the targeted cancer agent may be, for example, 10, 20, or 30 minutes, 1, 2, 4, 6, or 12 hours, or 1, 2, 3, 4, 5, 6 or 7 days or any period of time in this range. In certain embodiments, the anti-TIM-3 antibody molecule is administered multiple times over a period of time (e.g., 1, 2, 3, 4, 5, or 6 days, or 1, 2, 4, 8, 12, 16, or 20 weeks, or any period of time in this range) prior to administration of the targeted anti-cancer agent. In other embodiments, the anti-TIM-3 antibody molecule and the targeted cancer agent are administered substantially simultaneously.

Methods for the treatment of infectious diseases.

Other methods of the invention are used to treat patients who have been exposed to specific toxins or pathogens. Based on at least the Examples section of this disclosure, anti-TIM-3 antibodies can stimulate NK cell-mediated killing of target cells and can enhance IFN-gamma secretion and CD4+ T cell proliferation. Thus, in certain embodiments, the anti-TIM-3 antibody molecules described herein

- 73 040365 nii, are suitable for use in stimulating an immune response against an infectious agent. Thus, another aspect of the invention relates to a method of treating an infectious disease in a subject, comprising administering to the subject an anti-TIM-3 antibody molecule such that the subject is treated for the infectious disease. In the treatment of infection (eg, acute and/or chronic), administration of anti-TIM-3 antibody molecules can be combined with conventional treatments in addition to or instead of stimulating the host's natural immune defenses against infection. The host's natural immune defense against infection includes, but is not limited to, inflammation, fever, antibody-mediated host defense, T-lymphocyte-mediated host defense, including lymphokine secretion and cytotoxic T-cells (especially during viral infection), complement-mediated lysis, and opsonization. (facilitated by phagocytosis) and phagocytosis. The ability of anti-TIM-3 antibody molecules to reactivate dysfunctional T cells may be useful in the treatment of chronic infections, in particular infections in which cell-mediated immunity is essential for complete recovery.

Certain methods described herein are used to treat patients exposed to specific toxins or pathogens. In some aspects, a method of treating an infectious disease in a subject, comprising administering to the subject an anti-TIM-3 antibody molecule such that the subject is treated for the infectious disease.

Similar to use against tumors as discussed in the previous section, in some embodiments, anti-TIM-3 antibody molecules can be used alone or as an adjuvant, in combination with vaccines, to stimulate an immune response to, for example, pathogens or toxins. Examples of pathogens for which this therapeutic approach may be particularly useful include pathogens for which no effective vaccine currently exists or pathogens for which conventional vaccines are less than fully effective. These include, but are not limited to HIV, hepatitis (A, B and C), influenza virus, herpes virus, giardia, malaria, leishmania, Staphylococcus aureus, Pseudomonas Aeruginosa. Therapy with an anti-TIM-3 antibody molecule is also useful against advanced infections by agents such as HIV that present altered antigens during infections.

Thus, in some embodiments, an anti-TIM-3 antibody molecule is used to treat an individual who has an infection or is at risk of having an infection. Infection refers, for example, to a disease or condition due to the presence in the host of a foreign organism or an agent that reproduces in the host. Infections typically involve entry through the normal mucosa or other tissue barrier of an infectious organism or agent. An individual who has an infection is an individual having objectively measurable infectious organisms or agents present in the human body. An individual at risk of having an infection is an individual who is predisposed to developing an infection. Such an individual may include, for example, an individual with known or suspected exposure to an infectious organism or agent. An individual at risk of having an infection may also include an individual with a condition associated with a reduced ability to induce an immune response to an infectious organism or agent, for example, an individual with a congenital or acquired immunodeficiency, an individual undergoing radiation therapy or chemotherapy, an individual with brain damage, an individual with a traumatic injury, an individual undergoing surgery or other invasive medical or dental procedure.

Infections are broadly classified as bacterial, viral, fungal, or parasitic, based on the category of infectious organism or agent involved. Other lesser known types of infections include, for example, infections involving rickettsiae, mycoplasmas, and pruritus agents, bovine spongiform encephalopathy (BSE), and prion diseases (eg, kuru and Creutzfeldt-Jakob disease). Examples of bacteria, viruses, fungi and parasites that cause infection are well known in the art. The infection may be acute, subacute, chronic, or latent, and it may be localized or systemic. Moreover, the infection may be primarily intracellular or extracellular during at least one phase of the life cycle of the infectious organism or agent in the host.

Viruses

Examples of viruses found to cause infections in humans include, but are not limited to: Retroviridae (for example, human immunodeficiency viruses such as HIV-1 (also referred to as HTLV-III), HIV-2, LAV, or HTLV-III/LAV or HIV-III, and other isolates such as HIV-LP; Picornaviridae (eg, polioviruses, hepatitis A virus; enteroviruses, human coxsackieviruses, rhinoviruses, echoviruses); Calciviridae (eg, strains that cause gastroenteritis); Togaviridae (e.g. equine encephalitis viruses, rubella viruses); Flaviviridae (e.g. dengue viruses, encephalitis viruses, yellow fever viruses); Coronaviridae (e.g. coronaviruses); Rhabdoviridae (e.g. vesicular stomatitis viruses, rabies viruses); Filoviridae (e.g. Ebola viruses); Paramyxoviridae (e.g. parainfluenza viruses, mumps virus, measles virus,

- 74 040365 respiratory syncytial virus); Orthomyxoviridae (eg influenza viruses); Bungaviridae (eg hantaviruses, bunyaviruses, phleboviruses and nairoviruses); Arenaviridae (hemorrhagic fever viruses); Reoviridae (eg reoviruses, orbiviruses and rotaviruses); Birnaviridae; Hepadnaviridae (hepatitis B virus); Parvoviridae (parvoviruses); Papovaviridae (papilloma viruses, polyoma viruses); Adenoviridae (most adenoviruses); Herpesviridae (herpes simplex virus (HSV) 1 and 2, varicella zoster virus, cytomegalovirus (CMV), herpes virus; Poxyiridae (poxpox viruses, vaccinia viruses, poxviruses) and Iridoviridae (e.g., African swine fever virus); and unclassified viruses ( e.g. causative agents of spongiform encephalopathy, delta hepatitis agent (presumed to be a defective satellite of the hepatitis B virus), agents of neither A nor B hepatitis (class 1 = intestinally transmitted; class 2 = parenterally transmitted (i.e. hepatitis C); noroviruses and related viruses, and astroviruses). 1, HAV-6, HSV-II and CMV, Epstein-Barr virus), adenovirus, influenza virus, flaviviruses, echovirus, rhinovirus, coxsackievirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, in vaccinia virus, HTLV virus, dengue virus, human papillomavirus, molluscum virus, poliovirus, rabies virus, JC virus, and arboviral encephalitis virus.

For infections resulting from viruses, anti-TIM-3 antibody molecules can be combined through use simultaneously with, before or after standard therapies for treating viral infections. Such standard therapies vary depending on the type of virus, although in almost all cases it may be effective to administer human serum containing antibodies (eg, IgA, IgG) specific to the virus.

Some examples of pathogenic viruses that cause infections amenable to treatment include HIV, hepatitis (A, B, or C), herpes virus (e.g., VZV, HSV-1, HAV-6, HSV-II, and CMV, Epstein-Barr virus) , adenovirus, influenza virus, flaviviruses, echovirus, rhinovirus, coxsackievirus, coronovirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, mollusk virus, poliovirus, rabies virus, JC virus, arbovirus encephalitis virus and ebolaviruses (eg BDBV, EBOV, RESTV, SUDV and TAFV).

In one embodiment, the infection is an influenza virus infection. Influenza virus infection can cause fever, cough, myalgia, headache, and malaise, which often occur during seasonal epidemics. Influenza virus is also associated with a number of postinfectious disorders such as encephalitis, myopericarditis, Goodpasture's syndrome and Reye's syndrome. Influenza virus infection also suppresses the normal antibacterial defenses of the lungs, so that a patient recovering from influenza has an increased risk of developing bacterial pneumonia. The surface proteins of the virus show marked antigenic variation resulting from mutation and recombination. Thus, cytolytic T-lymphocytes are the host's primary means of eliminating the virus after infection. Influenza is classified into three main types: A, B, and C. The type A influenza virus is unique in that it infects both humans and many other animals (such as pigs, horses, birds, and seals) and is the main cause of pandemic influenza. Also, when a cell is infected with two different strains of the influenza A virus, the segmented RNA genomes of the two types of parental viruses mix during replication to form a hybrid replicant, resulting in new epidemic strains. Influenza B virus does not replicate in animals and thus has less genetic variation and influenza C virus has only one serotype.

Most conventional therapies are palliatives against symptoms resulting from infection, while in reality the disease is cleared by the host's immune response. However, certain strains (such as the influenza A virus) can cause more severe illness and death. Influenza A virus can be treated both clinically and prophylactically by administering the cyclic amine inhibitors amantadine and rimantidine, which inhibit viral replication. However, the clinical utility of these drugs is limited due to the relatively high incidence of adverse reactions that narrow the antiviral spectrum (influenza A virus only) and the tendency of the virus to become resistant. The introduction of serum IgG antibodies to the main surface proteins of the influenza virus: hemagglutinin and neuraminidase, can prevent pulmonary infection, while mucosal IgA is required to prevent infection of the upper respiratory tract and trachea. The most effective modern method of treating influenza is vaccination by introducing a virus inactivated with formalin or β-propiolactone.

In another embodiment, the infection is a hepatitis infection, such as hepatitis B or C infection.

Hepatitis B virus (HB-V) is the most infectious blood-borne pathogen. It is the main cause of acute and chronic hepatitis and liver carcinoma, as well as life

- 75 040365 chronic chronic infection. After infection, the virus replicates in hepatocytes, from which the surface antigen HBsAg is also separated. The detection of excessive serum HBsAg levels is used as a standard diagnostic tool for hepatitis B infection. An acute infection may resolve or may develop into a chronic persistent infection. Current treatments for chronic HBV include α-interferon, which upregulates human leukocyte antigen class I (HLA) expression on the surface of hepatocytes, thereby facilitating their recognition by cytotoxic T lymphocytes. In addition, the nucleoside analogues ganciclovir, famiciclovir and lamivudine have also shown some efficacy in treating HBV infection in clinical trials. Additional treatments for HBV include pegylated interferon-alpha, adenfovir, entecavir, and telbuvidin. Although passive immunity can be conferred through parenteral administration of serum antibodies against HBsAg, vaccination with inactivated or recombinant HBsAg also confers resistance to infection. Molecules of antibodies against T IM-3 can be combined with conventional methods for the treatment of hepatitis B infections to obtain a therapeutic advantage.

Infection with hepatitis C virus (HC-V) can lead to chronic hepatitis leading to cirrhosis. Although symptoms are similar to those of hepatitis B virus infection, unlike HB-V, infected hosts may be asymptomatic for 10 to 20 years. The anti-TIM-3 antibody molecule can be administered as monotherapy or in combination with standard treatment for hepatitis C infection. For example, the anti-TIM-3 antibody molecule can be administered with one or more of Sovaldi (sofosbuvir), Olisio (simeprevir), together with ribavirin or pegylated interferon. Although regimens that include Incivec (telaprevir) or Victrelis (boceprevir) plus ribavirin and pegylated interferon are also approved, they are associated with increased side effects and longer duration of treatment and thus are not considered preferred regimens.

The conventional method of treating HC-V infection involves administering a combination of α-interferon and ribavirin. A promising potential therapy for HC-V infection is the protease inhibitor telaprevir (VX-960). Additional treatments include: anti-PD-1 antibody (MDX-1106, Medarex), bavituximab (antibody that binds to the anionic phosphatidylserine phosphatidylserine in a B2-glycoprotein I dependent manner, Peregrine Pharmaceuticals), antibody(a) against viral envelope protein E2 HPV (eg ATL 6865-Ab68+Ab65, XTL Pharmaceuticals) and Civacir® (human anti-HCV polyclonal immunoglobulin). Anti-PD-L1 antibodies of the invention can be combined with one or more of these treatments for hepatitis C infections to provide a therapeutic benefit. Protease, polymerase and NS5A inhibitors that can be used in combination with anti-TIM-3 antibody molecules to specifically treat hepatitis C infection include those described in US 2013/0045202, incorporated herein by reference.

In another embodiment, the infection is a measles virus infection. After incubation for 9-11 days, hosts infected with the measles virus develop fever, cough, rhinitis, and conjunctivitis. Within 1-2 days, an erythematous mapopular rash develops, which quickly spreads throughout the body. Because infection also suppresses cellular immunity, the host is at higher risk of developing bacterial superinfections, including otitis media, pneumonia, and post-infectious encephalomyelitis. Acute infection is associated with significant morbidity and mortality, especially in malnourished adolescents.

Treatment of measles involves passive administration of pooled human IgG, which can prevent infection in non-immune individuals, even when administered up to one week after exposure. However, prior immunization with live attenuated virus is the most effective treatment and prevents disease in more than 95% of immunized individuals. Since there is one serotype of the virus, one immunization or infection usually results in lifelong protection against subsequent infection.

In a small proportion of infected hosts, measles can develop into SSPE, which is a chronic, progressive neurological disorder resulting from persistent infection of the central nervous system. SSPE is caused by clonal variants of the measles virus with defects that prevent assembly and budding of the virus. For these patients, it may be desirable to reactivate T cells with anti-TIM-3 antibody molecules to promote clearance of the virus.

In another embodiment, the infection is HIV. HIV attacks CD4 ⁺ cells, including T lymphocytes, macrophage monocytes, follicular dendritic cells, and Langerhans cells, and CD4+ helper/inducing cells are depleted. As a result, the host acquires a severe defect in cell-mediated immunity. HIV infection leads to AIDS in at least 50% of individuals and is transmitted through sexual contact, ingestion of infected blood or blood products, artificial insemination with infected semen, exposure to blood-containing needles or syringes, and from an infected mother to an infant during childbirth.

- 76 040365

An HIV-infected host may be asymptomatic, or may develop an acute illness that resembles mononucleosis—fever, headache, sore throat, malaise, and rash. Symptoms may progress to progressive immune dysfunction, including persistent fever, night sweats, weight loss, unexplained diarrhea, eczema, psoriasis, seborrheic dermatitis, herpes zoster, oral candidiasis, and oral hairy leukoplakia. In patients whose infections develop into AIDS, there are frequent opportunistic host infections with parasites.

HIV treatment includes antiviral therapy, including nucleoside analogs, zidovudine (AST), either alone or in combination with didanosine or zalcitabine, dideoxyinosine, dideoxycytidine, lamivudine, stavudine; reverse transcriptase inhibitors such as delavirdine, nevirapine, loviride; and protease inhibitors such as saquinavir, ritonavir, indinavir and nelfinavir. Anti-TIM-3 antibody molecules can be combined with conventional treatments for HIV infections to provide a therapeutic benefit.

In another embodiment, the infection is a cytomegalovirus (CMV) infection. CMV infection is often associated with persistent, latent, and recurrent infection. CMV infects monocytes and monocyte granulocyte progenitors and remains latent in them. Clinical symptoms of CMV include mononucleosis-like symptoms (ie, fever, swollen tonsils, malaise) and a tendency to develop an allergic skin rash to antibiotics. The virus is spread through direct contact. The virus is excreted in urine, saliva, semen and, to a lesser extent, other body fluids. Transmission can also occur from an infected mother to her fetus or newborn child and through blood transfusion and organ transplantation. Infection with CMV typically results in a decrease in cellular immunity characterized by decreased blastogenic responses to non-specific mitogens and specific CMV antigens, decreased cytotoxic capacity, and increased CD8 or CD4+ lymphocyte counts.

Treatment for CMV infection includes the antiviral drugs ganciclovir, foscarnet, and sidovir, but these drugs are generally reserved for immunocompromised patients. Anti-TIM-3 antibody molecules can be combined with conventional treatments for cytomegalovirus infections to provide a therapeutic benefit.

In another embodiment, the infection is an Epstein Barr virus (EBV) infection. EBV can cause persistent and latent infections and mainly attacks B cells. Infection with EBV results in the clinical condition of infectious mononucleosis, which includes fever, sore throat often with exudate, generalized lymphadenopathy, and splenomegaly. Hepatitis is also present, as a result of which jaundice may develop.

Although palliative treatment of symptoms is the typical treatment for EBV infections, EBV has been associated with the development of certain malignancies such as Burkitt's lymphoma and nasopharyngeal cancer. Thus, clearing the viral infection before these complications occur would be of significant benefit. Anti-TIM-3 antibody molecules can be combined with conventional treatments for Epstein-Barr virus infections to provide a therapeutic benefit.

In another embodiment, the infection is a herpes simplex virus (HSV) infection. HSV is transmitted through direct contact with an infected host. Direct infection may be asymptomatic, but usually results in bullous blisters containing infectious particles. The disease manifests itself as cycles of active periods of the disease, in which lesions appear and disappear as the virus latently infects the nerve ganglion for subsequent exacerbations. Lesions may be on the face, genitals, eyes and/or hands. In some cases, the infection can also cause encephalitis.

Treatments for herpes virus infections are primarily aimed at resolving symptomatic exacerbations and include systemic antiviral drugs such as: acyclovir (eg Zovirax®), valaciclovir, famiciclovir, penciclovir, and topical drugs such as provenol (Abreva®), tromantidin and silactin. The elimination of latent infections with the herpes virus would be of significant clinical benefit. Anti-TIM-3 antibody molecules can be combined with conventional treatments for herpes virus infections to provide a therapeutic benefit.

In another embodiment, the infection is a human T-lymphotropic virus (HTLV-1, HTLV-2) infection. HTLV is transmitted through sexual contact, breastfeeding, or exposure to contaminated blood. The virus activates a subset of TH cells called Th1 cells, resulting in overproliferation and overproduction of Th1 cytokines (eg, IFN-γ and TNF-α). This in turn leads to suppression of Th2 lymphocytes and reduced production of Th2 cytokines (e.g., IL-4, IL-5, IL-10, and IL-13), causing a decrease in the ability of the infected host to induce an adequate immune response to invading organisms, for the elimination of which a Th2-dependent response is required (eg, parasitic infections, production of mucosal and humoral

- 77 040365 ral antibodies).

HTLV infections lead to opportunistic infections causing bronchiectasis, dermatitis, and superinfections with Staphylococcus spp. and Strongyloides spp., resulting in death from polymicrobial sepsis. HTLV infection can also lead directly to adult T-cell leukemia/lymphoma and progressive upper motor neuron demyelinating disease known as HAM/TSP. Clearing latent HTLV infections may have significant clinical benefit. Anti-TIM-3 antibody molecules can be combined with conventional treatments for HTLV infections to provide a therapeutic benefit.

In another embodiment, the infection is a human papillomavirus (HPV) infection. HPV mainly affects keratinocytes and occurs in two forms: cutaneous and genital. Transmission is believed to occur through direct contact and/or sexual activity. Both skin and genital HPV infection can lead to warts and latent infections and, sometimes, recurrent infections that are controlled by the host's immune system, which controls symptoms and blocks the appearance of warts, but leaves the host able to transmit the infection to others.

HPV infection can also lead to certain cancers such as cancer of the cervix, anal canal, vulva, penis, and oropharynx. There are currently no known treatments for HPV infection, however, the current treatment is topical imiquimod, which stimulates the immune system to attack the affected area. Clearing latent HPV infections can be of significant clinical benefit. Anti-TIM-3 antibodies of the invention can be combined with conventional treatments for HPV infections to provide a therapeutic benefit.

In another embodiment, the infection is an Ebola virus (EBOV). EBOV is one of the five known Ebolaviruses. EBOV causes a severe and often fatal hemorrhagic fever in humans and mammals, known as Ebola virus disease (EVD). Transmission occurs through contact with the blood, secretions, organs, or other body fluids of infected patients. There is currently no approved treatment or vaccine.

bacterial infections.

Bacteria include Gram-negative and Gram-positive bacteria. Examples of Gram-positive bacteria include, but are not limited to, Pasteurella spp., Staphylococcus spp., and Streptococcus spp. Examples of Gram-negative bacteria include, but are not limited to, Escherichia coli, Pseudomonas species, and Salmonella species. Specific examples of infectious bacteria include, but are not limited to: Helicobacter pyloris, Borrelia burgdorferi, Legionella pneumophilia, Mycobacteria spp. (e.g., M. tuberculosis, M. avium, M. intracellulare, M. kansasii, M. gordonae), Staphylococcus aureus, Neisseria gonorrhoeae, Neisseria meningitidis, Listeria monocytogenes, Streptococcus pyogenes (group A streptococcus), Streptococcus agalactiae (group B streptococcus ), Streptococcus (viridans group), Streptococcus faecalis, Streptococcus bovis, Streptococcus (anaerobic species.), Streptococcus pneumoniae, pathogenic Campylobacter spp., Enterococcus spp., Haemophilus influenzae, Bacillus anthracis, Corynebacterium diphtheriae, Corynebacterium spp., Erysipelothrix rhustiopathiae perfringens, Clostridium tetani, Enterobacter aerogenes, Klebsiella pneumoniae, Pasturella multocida, Bacteroides spp., Fusobacterium nucleatum, Streptobacillus moniliformis, Treponema pallidum, Treponema pertenue, Leptospira, Mycobacterium leprae, Rickettsia and Actinomyces israelii. Some examples of pathogenic bacteria that cause infections amenable to treatment by the methods described herein include Chlamydia, Rickettsial bacteria, Mycobacteria, Staphylococci, Streptococci, Pneumococci, Meningococci and Gonococci, Klebsiella, Proteus, Serratia, Pseudomonas, Legionella, Diphtheria, Salmonella, bacilli, cholera pathogens, tetanus pathogens, botulism pathogens, anthrax pathogens, plague pathogens, leptospirosis pathogens and Lyme disease pathogens.

Some examples of pathogenic bacteria that cause infections treatable by the methods of the invention include syphilis, chlamydia, rickettsial bacteria, mycobacteria, staphylococci, streptococci, pneumococci, meningococci and gonococci, Klebsiella, Proteus, serratia, Pseudomonas, Legionella, diphtheria, salmonella, bacilli, cholera pathogens, tetanus pathogens, botulism pathogens, anthrax pathogens, plague pathogens, leptospirosis pathogens and Lyme disease pathogens. Anti-TIM-3 antibody molecules can be used in combination with existing treatments for the above infections. For example, treatments for syphilis include penicillin (eg, penicillin G), tetracycline, doxycillin, ceftriaxone, and azithromycin.

Lyme disease, caused by Borrelia burgdorferi, is transmitted to humans through tick bites. The disease initially presents as a localized rash followed by flu-like symptoms. Including malaise, fever, headache, neck stiffness and joint pain. Later manifestations may include migratory and polyarticular arthritis, nervous system and cardiac involvement with cranial nerve palsies and radiculopathy, myocarditis, and arrhythmias. Not

- 78 040365 which cases of Lyme disease become persistent, causing irreversible damage similar to tertiary syphilis. Current treatments for Lyme disease mainly involve the administration of antibiotics. Antibiotic-resistant strains can be treated with hydroxychloroquine or methotrexate. Antibiotic-refractory patients with neuropathic pain can be treated with gabapentin. Minocycline may be useful in late/chronic Lyme disease with neurological or other inflammatory manifestations.

Other forms of borreliosis, such as those resulting from infection with B. recurentis, B. hermsii, B. turicatae, B. parikeri, B. hispanica, B. duttonii, and B. persica, and leptospirosis (eg, L. interrogans) , as a rule, are resolved spontaneously if the titers in the blood do not reach a concentration that causes intrahepatic obstruction.

fungi and parasites.

Examples of fungi include: Aspergillus spp., Blastomyces dermatitidis, Candida albicans, other Candida spp., Coccidioides immitis, Cryptococcus neoformans, Histoplasma capsulatum, Chlamydia trachomatis, Nocardia spp., Pneumocystis carinri. Some examples of pathogenic fungi that cause infections treatable by the methods described herein include Candida (albicans, krusei, glabrata, tropicalis, etc.), Cryptococcus neoformans, Aspergillus (fumigatus, niger, etc.), genus Mucorales (mucor, absidia, rhizophus), Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis and Histoplasma capsulatum.

Parasites include, but are not limited to, blood-borne and/or tissue-borne parasites such as Babesia microti, Babesia divergens, Entamoeba histolytica, Giardia lamblia, Leishmania tropica, Leishmania spp., Leishmania braziliensis, Leishmania donovani, Plasmodium falciparum, Plasmodium malariae, Plasmodium ovale, Plasmodium vivax and Toxoplasma gondii, Trypanosoma gambiense and Trypanosoma rhodesiense (African sleeping sickness), Trypanosoma cruzi (Chagas disease) and Toxoplasma gondii, flatworms, roundworms. Some examples of pathogenic parasites that cause infections amenable to treatment by the methods described herein include Entamoeba histolytica, Balantidium coli, Naegleriafowleri, Acanthamoeba sp., Giardia lambia, Cryptosporidium sp., Pneumocystis carinii, Plasmodium vivax, Babesia microti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani, Toxoplasma gondii and Nippostrongylus brasiliensis.

Some examples of pathogenic fungi that cause infections treatable by the methods of the invention include Candida (albicans, krusei, glabrata, tropicalis, etc.), Cryptococcus neoformans, Aspergillus (fumigatus, niger, etc.), Genus Mucorales (mucor , absidia, rhizophus), Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis and Histoplasma capsulatum.

Some examples of pathogenic parasites that cause infections amenable to treatment by the methods described herein include Entamoeba histolytica, Balantidium coli, Naegleriafowleri, Acanthamoeba sp., Giardia lambia, Cryptosporidium sp., Pneumocystis carinii, Plasmodium vivax, Babesia microti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani, Toxoplasma gondii and Nippostrongylus brasiliensis.

In some embodiments, the infectious disease is selected from hepatitis (eg, hepatitis C virus infection) or sepsis.

In all of the methods described above, therapy with an anti-TIM-3 antibody molecule can be combined with other forms of immunotherapy, such as treatment with cytokines (e.g., interferons, GM-CSF, G-CSF, IL-2, IL-21) or bispecific antibody therapy, which provides enhanced presentation of tumor antigens (see, for example, Holliger (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak (1994) Structure 2:1121-1123).

Methods for introducing various antibody molecules are known in the art and are described below. Suitable dosages of antibody molecules depend on the age and body weight of the individual and the particular drug used. Antibody molecules can be used as competitive means to bind a ligand to inhibit or reduce an undesirable interaction.

The antibody molecules can be used alone or in the form of a conjugate with a second agent, such as a cytotoxic drug, a radioisotope, or a protein, such as a protein toxin or a viral protein. This method includes: administering an antibody molecule, alone or conjugated to a cytotoxic drug, to an individual in need of such treatment. Antibody molecules can be used to deliver various drugs, e.g., a cytotoxic moiety, e.g., a therapeutic drug, radioisotopes, molecules of plant, fungal, or bacterial origin, or biological proteins (e.g., protein toxins) or particles (e.g., recombinant viral particles, e.g.; through the viral envelope protein) or mixtures thereof.

Additional combined therapies.

Anti-TIM-3 antibody molecules can be used in combination with other therapies. For example, the combination therapy may include an anti-TIM-3 antibody molecule co-formulated and/or co-administered with one or more additional drugs, e.g., one or more anti-cancer agents, cytotoxic

- 79 040365 chemical or cytostatic agents, hormonal treatments, vaccines and/or other immunotherapies. In other embodiments, the antibody molecules are administered in combination with other therapies, including surgery, radiation therapy, cryosurgery, and/or thermotherapy. Such combination therapies can advantageously utilize lower dosages of administered drugs, thus avoiding the potential toxicity or complications associated with different monotherapies.

Administration in combination, as used herein, means that two (or more) different drugs are delivered to an individual during the period of time that the individual is suffering from the disorder, e.g., two or more drugs are delivered after the individual has been diagnosed with a disorder and until the individual is cured or eliminated. violations. In some embodiments, the implementation of one method of treatment is still in progress when the implementation of the second method begins, so that there is an overlap. This is sometimes referred to herein as simultaneous or co-delivery. In other embodiments, delivery of one drug ends before delivery of another drug begins. In some embodiments, the implementation of any of these cases, the treatment is more effective due to the combined introduction. For example, the second drug is more effective, e.g., an equivalent effect is observed using less of the second drug, or the second drug reduces symptoms more than would be observed if the second drug was administered in the absence of the first drug, or the like. the situation is observed for the first drug. In some embodiments, delivery is such that the reduction in a symptom or other parameter associated with a disorder is greater than would be observed when one drug is delivered in the absence of the other. The effect of the two drugs may be partially additive, fully additive, or more than additive. The delivery may be such that the effect of the first drug delivered is still detectable when the second drug is delivered.

Molecules of antibodies against TIM-3 can be administered in combination with one or more existing methods of treating malignant tumors, including, but not limited to: surgery; radiation therapy (for example, external beam therapy, which involves three-dimensional conformal radiation therapy, where the area of radiation is calculated).

In some aspects, the anti-TIM-3 antibody is co-administered with a second agent that acts on TIM-3 or another element of the TIM-3 cascade.

In some embodiments, for example in the treatment of an infectious disease, the TIM-3 antibody can be co-administered with, for example, an antibiotic, antiviral agent, or antifungal agent.

In some embodiments, for example in the treatment of Crohn's disease, the anti-TIM-3 antibody can be co-administered with, for example, an anti-inflammatory drug such as 5-aminosalicylic acid (5-ASA), prednisone, or hydrocortisone; purine analogues such as azathioprine; antimetabolites such as methotrexate; TNF-alpha inhibitors, eg anti-tumor necrosis factor alpha (TNF-α) monoclonal antibody, eg infliximab, adalimumab or certolizumab; or integrin inhibitors, such as an anti-alpha-4 integrin monoclonal antibody, such as natalizumab.

In some embodiments, for example in the treatment of multiple sclerosis, the anti-TIM-3 antibody can be co-administered with, for example, an interferon such as interferon beta-1a, interferon beta-1b, an interferon analog, a random amino acid polymer such as glatiramer acetate; a type II topoisomerase inhibitor such as mitoxantrone; an integrin inhibitor, such as an anti-alpha-4 integrin monoclonal antibody, such as natalizumab; sphingosine-1 phosphate receptor modulator, eg fingolimod; a pyrimidine synthesis inhibitor, for example a dihydroorotate dehydrogenase inhibitor such as teriflunomide; and other immunomodulatory agents such as dimethyl fumarate.

In some embodiments, for example in the treatment of sepsis, the anti-TIM-3 antibody can be co-administered with, for example, antibiotics; vasoconstrictor factors such as norepinephrine or dopamine; steroids; recombinant activated protein C (drotrecogin alfa); intravenous fluids and external respiration.

In some embodiments, for example in the treatment of SIRS (Systemic Inflammatory Response Syndrome), the anti-TIM-3 antibody can be co-administered with, for example, antibiotics; steroids; antioxidants or intravenous fluids.

In some embodiments, for example in the treatment of glomerulonephritis, the anti-TIM-3 antibody can be co-administered with, for example, steroids; an alkylating agent such as cyclophosphamide; or a purine analogue such as azathioprine.

Combinations of anti-TIM-3 antibody molecules with one or more second therapeutic agents are contemplated within the scope of the present invention. Many of the combinations in this section

- 80 040365 are useful in the treatment of cancer, but other indications are also described. This section focuses on combinations of anti-TIM-3 antibody molecules, optionally in combination with one or more immunomodulators (e.g., anti-PD-1 antibody molecule, anti-LAG-3 antibody molecule, or anti-PD-L1 antibody molecule), with one or more agents. described in Table. 6. In the combinations described below, in one embodiment, the anti-TIM-3 antibody molecule comprises (i) a heavy chain variable region (VH) comprising a VHCDR1 amino acid sequence selected from SEQ ID NO: 3 or SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 4, SEQ ID NO: 10, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 30 or SEQ ID NO: 31 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and (ii) a light chain variable region (VL) comprising the VLCDR1 amino acid sequence of SEQ ID NO: 6 or SEQ ID NO: 12, the VLCDR2 amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 13, and the VLCDR3 amino acid sequence of SEQ ID NO: 8 or SEQ ID NO: 14.

In one embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with the PKC inhibitor sotrastaurine (Compound A1) or a compound described in PCT Publication No. WO 2005/039549 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the PKC inhibitor is described in table. 6 or in the publication given in table. 6, for example, in row A1 of Table. 6. In one embodiment, the PKC inhibitor is sotrastaurine (Compound A1) or the compound described in PCT Publication No. WO 2005/039549. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with sotrastaurine (Compound A1) or a compound as described in PCT Publication No. WO 2005/039549 to treat a disorder such as cancer, melanoma, non-Hodgkin's lymphoma, inflammatory bowel disease , transplant rejection, ophthalmic disorder, or psoriasis.

In certain embodiments, sotrastaurine (Compound A1) is administered at a dose of about 20 to 600 mg, such as about 200 to about 600 mg, about 50 mg to about 450 mg, about 100 to 400 mg, about 150 to 350 mg or about 200 to 300 mg, such as about 50 mg, 100 mg, 150 mg, 200 mg, 300 mg, 400 mg, 500 or 600 mg. The dosing regimen may vary, for example, from administration every other day to administration every day, twice or three times a day.

In one embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with the BCR-ABL inhibitor TASIGNA (compound A2 or compound described in PCT Publication No. WO 2004/005281) for the treatment of a disorder, such as the disorder described herein. In one embodiment, the BCR-ABL inhibitor is TASIGNA or a compound described in PCT Publication No. WO 2004/005281. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with TASIGNA (Compound A2) or a compound as described in PCT Publication No. WO 2004/005281 for the treatment of a disorder such as lymphocytic leukemia, Parkinson's disease, neurological malignancy, melanoma , gastrointestinal/gastrointestinal cancer, colorectal cancer, myeloid leukemia, head and neck cancer, or pulmonary hypertension.

In one embodiment, the BCR-ABL inhibitor or TASIGNA is administered at a dose of about 300 mg (e.g., twice daily, e.g., for newly diagnosed Ph+ CML-CP) or about 400 mg, e.g., twice daily, e.g., for resistant or intolerant Ph+ CML-CP and CML-AP). The BCR-ABL inhibitor or Compound A2 is administered at a dose of approximately 300-400 mg.

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an HSP90 inhibitor such as 5-(2, 4-dihydroxy-5-isopropylphenyl)-N-ethyl-4-(4-(morpholinomethyl)phenyl)isoxazole-3-carboxamide (Compound A3) or a compound described in PCT Publication No. WO 2010/060937 or WO 2004/072051, for treatment violations, such as violations described in the present description. In one embodiment, the HSP90 inhibitor is 5-(2,4-dihydroxy-5-isopropylphenyl)-N-ethyl-4-(4(morpholinomethyl)phenyl)isoxazole-3-carboxamide (compound A3) or a compound described in the publication PCT No. WO 2010/060937 or WO 2004/072051. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with 5-(2,4-dihydroxy-5-isopropylphenyl)-N-ethyl-4-(4(morpholinomethyl)phenyl)isoxazole-3-carboxamide (compound A3) or a compound as described in PCT Publication No. WO 2010/060937 or WO 2004/072051 for the treatment of a disorder such as cancer, multiple myeloma, non-small cell lung cancer, lymphoma, gastric cancer, breast cancer, gastrointestinal/gastrointestinal cancer. intestinal cancer, pancreatic cancer, colorectal cancer, solid tumor or hematopoietic disorder.

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more

- 81 040365 other immunomodulators, used in combination with the PI3K inhibitor and/or mTOR dactolisib (Compound A4) or 8-(6-methoxypyridin-3-yl)-3-methyl-1-(4-piperazin-1-yl-3trifluoromethylphenyl )-1,3-dihydroimidazo[4,5-c]quinolin-2-one (Compound A41) or the compound described in PCT Publication No. WO 2006/122806 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the PI3K and/or mTOR inhibitor is dactolisib (compound A4), 8-(6-methoxypyridin-3-yl)-3-methyl-1-(4-piperazin-1-yl-3trifluoromethylphenyl)-1, 3-dihydroimidazo[4,5-c]quinolin-2-one (Compound A41) or the compound described in PCT Publication No. WO 2006/122806. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with dactolisib (compound A4), 8-(6-methoxypyridin-3-yl)-3methyl-1-(4-piperazin-1-yl-3-trifluoromethylphenyl)- 1,3-dihydroimidazo[4,5-c]quinolin-2-one (Compound A41) or the compound described in PCT Publication No. WO 2006/122806 for the treatment of a disorder such as cancer, prostate cancer, leukemia (e.g. , lymphocytic leukemia), breast cancer, brain cancer, bladder cancer, pancreatic cancer, kidney cancer, solid tumor, or liver cancer.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an FGFR 3-(2,6-dichloro -3,5dimethoxyphenyl)-1-(6-((4-(4-ethylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-1-methylurea (compound A5) or the compound described in US Pat. No. 8,552,002 , for the treatment of a disorder, such as the disorder described herein. In one embodiment, the FGFR inhibitor is 3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-(6-((4-(4-ethylpiperazin-1-yl)phenyl)amino)pyrimidin-4- yl)-1-methylurea (Compound A5) or the compound described in US Pat. No. 8,552,002. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with Compound A5 or a compound as described in US Pat. gastrointestinal cancer/gastrointestinal cancer, hematologic malignancy or solid tumor.

In one embodiment, an FGFR inhibitor or 3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-(6-((4(4-ethylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl) -1-methylurea (Compound A5) is administered at a dose of about 100-125 mg (eg per day), eg about 100 mg or about 125 mg.

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with the PI3K inhibitor buparlisib (Compound A6) or a compound described in PCT Publication No. WO 2007/084786 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the PI3K inhibitor is buparlisib (Compound A6) or the compound described in PCT Publication No. WO 2007/084786. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with buparlisib (compound A6) or a compound described in PCT Publication No. WO 2007/084786 to treat a disorder such as prostate cancer, non-small cell lung cancer, endocrine malignancy, leukemia, ovarian cancer, melanoma, bladder cancer, breast cancer, cancer of the female reproductive system, digestive tract/gastrointestinal cancer, colorectal cancer, glioblastoma multiforme, solid tumor, non-Hodgkin's lymphoma, hematopoietic disorder, or head cancer, and neck.

In one embodiment, the PI3K inhibitor or buparlisib (compound A6) is administered at a dose of about 100 mg (eg, per day).

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an FGFR inhibitor 8-(2,6-difluoro -3,5dimethoxyphenyl)-N-(4-((dimethylamino)methyl)-1H-imidazol-2-yl)quinoxalin-5-carboxamide (Compound A7) or the compound described in PCT Publication No. WO 2009/141386 for treatment violations, such as violations described in the present description. In one embodiment, the FGFR inhibitor is 8-(2,6-difluoro-3,5-dimethoxyphenyl)-N-(4-((dimethylamino)methyl)-1H-imidazol-2-yl)quinoxalin-5-carboxamide ( compound A7) or the compound described in PCT Publication No. WO 2009/141386. In one embodiment, the FGFR inhibitor is 8-(2,6difluoro-3,5-dimethoxyphenyl)-N-(4-((dimethylamino)methyl)-1H-imidazol-2-yl)quinoxalin-5-carboxamide (compound A7 ). In one embodiment, an anti-TIM-3 antibody molecule is used in combination with 8-(2,6-difluoro-3,5-dimethoxyphenyl)-N-(4-((dimethylamino)methyl)-1H-imidazol-2-yl) quinoxaline-5-carboxamide (Compound A7) or the compound described in PCT Publication No. WO 2009/141386 for the treatment of a disorder such as cancer characterized by angiogenesis.

In one embodiment, an FGFR inhibitor or 8-(2,6-difluoro-3,5-dimethoxyphenyl)-K-(4((dimethylamino)methyl)-1H-imidazol-2-yl)quinoxalin-5-carboxamide (compound A7 ) is administered at a dose

- 82 040365 for example, from about 3 mg to about 5 g, more preferably from about 10 mg to about 1.5 g per individual per day, optionally divided into 1-3 separate doses, which may have, for example, the same size.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a PI3K(S)-N1-(4 -methyl-5-(2(1,1,1-trifluoro-2-methylpropan-2-yl)pyridin-4-yl)thiazol-2-yl)pyrrolidin-1,2-dicarboxamide (compound A8) or a compound, described in PCT Publication No. WO 2010/029082 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the PI3K inhibitor is (S)-N1-(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropan-2-yl)pyridin-4-yl)thiazol-2 -yl)pyrrolidine-1,2-dicarboxamide (Compound A8) or the compound described in PCT Publication No. WO 2010/029082. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with (S)-N 1 -(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropan-2-yl)pyridin- 4-yl)thiazol-2-yl)pyrrolidin-1,2-dicarboxamide (Compound A8) or the compound described in PCT Publication No. WO 2010/029082 for the treatment of a disorder such as gastric cancer, breast cancer, pancreatic cancer , gastrointestinal/gastrointestinal cancer, solid tumor, and head and neck cancer.

In one embodiment, a PI3K inhibitor or (S)-N1-(4-methyl-5-(2-(1,1,1-trifluoro-2methylpropan-2-yl)pyridin-4-yl)thiazol-2-yl) pyrrolidine-1,2-dicarboxamide (Compound A8) is administered at a dose of about 150-300, 200-300, 200-400, or 300-400 mg (eg, per day), for example, about 200, 300, or 400 mg.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a cytochrome P450 inhibitor (e.g., a CYP17 inhibitor), or a compound described in PCT Publication No. WO 2010/149755 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the cytochrome P450 inhibitor (eg, CYP17 inhibitor) is a compound described in PCT Publication No. WO 2010/149755. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with a compound described in PCT Publication No. WO 2010/149755 for the treatment of prostate cancer.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an HDM2(S)-1-(4 -chlorophenyl)-7isopropoxy-6-methoxy-2-(4-(methyl(((1r,4S)-4-(4-methyl-3-oxopiperazin-1-yl)cyclohexyl)methyl)amino)phenyl)-1 ,2-dihydroisoquinolin-3(4H)-one (compound A10) or the compound described in PCT Publication No. WO 2011/076786 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the HDM2 inhibitor is (S)-1-(4-chlorophenyl)7-isopropoxy-6-methoxy-2-(4-(methyl(((1r,4S)-4-(4-methyl-3 -oxopiperazin-1-yl)cyclohexyl)methyl)amino)phenyl)-1,2-dihydroisoquinolin-3(4H)-one (Compound A10) or the compound described in PCT Publication No. WO 2011/076786. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with (S)-1-(4-chlorophenyl)-7-isopropoxy-6-methoxy-2-(4-(methyl(((1r,4S)-4 -(4-methyl-3-oxopiperazin-1-yl)cyclohexyl)methyl)amino)phenyl)-1,2-dihydroisoquinolin-3(4H)-one (compound A10) or the compound described in PCT Publication No. WO 2011/ 076786, for the treatment of a disorder such as a solid tumor.

In one embodiment, an HDM2 inhibitor or (S)-1-(4-chlorophenyl)-7-isopropoxy-6methoxy-2-(4-(methyl(((1r,4S)-4-(4-methyl-3-oxopiperazine -1-yl)cyclohexyl)methyl)amino)phenyl)-1,2dihydroisoquinolin-3(4H)-one (compound A10) is administered at a dose of approximately 400 to 700 mg, for example, administered three times a week, two 2 weeks from administration and one week without administration. In some embodiments, the dose is about 400, 500, 600, or 700 mg; about 400-500, 500-600 or 600-700 mg, for example, administered three times a week.

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with the iron chelating agent deferasirox (also known as EXJADE; compound A11) or a compound described in PCT Publication No. WO 1997/049395 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the iron chelating agent is deferasirox or the compound described in PCT Publication No. WO 1997/049395. In one embodiment, the iron chelating agent is deferasirox (compound A11). In one embodiment, an anti-TIM-3 antibody molecule is used in combination with deferasirox (Compound A11) or a compound described in PCT Publication No. WO1997/049395 to treat iron overload, hemochromatosis, or myelodysplasia.

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more

- 83 040365 other immunomodulators are used in combination with the aromatase inhibitor lectrozole (also known as FEMARA; compound A12) or a compound described in US 4978672 to treat a disorder, for example the disorder described herein. In one embodiment, the aromatase inhibitor is Lectrosol (Compound A12) or a compound described in US Pat. No. 4,978,672. for the treatment of a disorder such as cancer, leiomyosarcoma, endometrial cancer, breast cancer, cancer of the female reproductive system, or hormone deficiency.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a PI3K inhibitor, e.g., a common PI3K (4S, 5R)-3-(2'-amino-2-morpholino-4'-(trifluoromethyl)-[4,5'-bipyrimidin]-6-yl)-4-(hydroxymethyl)-5-methyloxazolidin-2-one ( compound A13) or a compound described in PCT Publication No. WO 2013/124826 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the PI3K inhibitor is (4S,5R)-3-(2'-amino-2-morpholino-4'(trifluoromethyl)-[4,5'-bipyrimidin]-6-yl)-4-(hydroxymethyl )-5-methyloxazolidin-2-one (Compound A13) or the compound described in PCT Publication No. WO 2013/124826. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with (4S,5R)-3-(2'-amino-2-morpholino-4'(trifluoromethyl)-[4,5'-bipyrimidin]-6-yl )-4-(hydroxymethyl)-5-methyloxazolidin-2-one (Compound A13) or the compound described in PCT Publication No. WO 2013/124826 for the treatment of a disorder such as a malignant tumor or an advanced solid tumor.

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an inhibitor of p53 and/or p53/Mdm2 interaction ( S)-5-(5-chloro-1-methyl-2-oxo-1,2-dihydropyridin-3-yl)-6-(4-chlorophenyl)-2-(2,4-dimethoxypyrimidin-5-yl) -1-isopropyl-5,6-dihydropyrrolo[3,4-d]imidazol-4(1H)-one (Compound A14) or a compound described in PCT Publication No. WO 2013/111105 for the treatment of a disorder, for example the disorder described in the present description. In one embodiment, the p53 and/or p53/Mdm2 interaction inhibitor is (S)-5-(5-chloro-1-methyl-2-oxo-1,2-dihydropyridin-3-yl)6-(4-chlorophenyl )-2-(2,4-dimethoxypyrimidin-5-yl)-1-isopropyl-5,6-dihydropyrrolo[3,4-d]imidazol4(1H)-one (compound A14) or the compound described in PCT Publication No. WO 2013/111105. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with (S)-5-(5-chloro-1methyl-2-oxo-1,2-dihydropyridin-3-yl)-6-(4-chlorophenyl)- 2-(2,4-dimethoxypyrimidin-5-yl)-1-isopropyl-5,6-dihydropyrrolo[3,4-d]imidazol-4(1H)-one (compound A14) or the compound described in PCT Publication No. WO 2013/111105 for the treatment of a disorder such as cancer or soft tissue sarcoma.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a CSF-1R tyrosine kinase inhibitor 4-((2 (((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15) or the compound described in PCT Publication No. WO 2005/073224 for the treatment of the disorder , such as the violation described in the present description. In one embodiment, the CSF-1R tyrosine kinase inhibitor is 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol6-yl)oxy)-N-methylpicolinamide (compound A15) or the compound described in PCT Publication No. WO 2005/073224. In one embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, is used in combination with 4-((2-(((1R,2R)2-hydroxycyclohexyl)amino)benzo[ d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15) or the compound described in PCT Publication No. WO 2005/073224 for the treatment of a disorder such as cancer.

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an apoptosis inducer and/or an angiogenesis inhibitor, such as imatinib mesylate (also known as GLEEVEC; compound A16), or the compound described in PCT Publication No. WO1999/003854, for the treatment of a disorder, eg, the disorder described. In one embodiment, the apoptosis inducer and/or angiogenesis inhibitor is imatinib mesylate (Compound A16) or the compound described in PCT Publication No. WO1999/003854. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with imatinib mesylate (Compound A16) or a compound described in PCT Publication No. WO1999/003854 to treat a disorder such as cancer, multiple myeloma, prostate cancer, non-small cell cancer. lung, lymphoma, gastric cancer, melanoma, breast cancer, pancreatic cancer, gastrointestinal/gastrointestinal cancer, colorectal cancer, glioblastoma multiforme, liver cancer, head and neck cancer, asthma, race

- 84 040365 multiple sclerosis, allergies, dementia of the Alzheimer's type, amyotrophic lateral sclerosis or rheumatoid arthritis.

In certain embodiments, imatinib mesylate (compound A16) is administered at a dose of about 100 to 1000 mg, such as about 200 mg to 800 mg, about 300 mg to 700 mg, or about 400 mg to 600 mg, such as about 200 mg, 300 mg, 400 mg, 500 mg, 600 mg or 700 mg. The dosing regimen may vary from, for example, once every two days to once a day, two or three times a day. In one embodiment, imatinib mesylate is administered at an oral dose of about 100 mg to 600 mg every day, such as about 100 mg, 200 mg, 260 mg, 300 mg, 400 mg, or 600 mg every day.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a JAK inhibitor 2-fluoro-N-methyl- 4-(7(quinolin-6-ylmethyl)imidazo[1,2-b][1,2,4]triazin-2-yl)benzamide (compound A17) or dihydrochloride thereof or with the compound described in PCT Publication No. WO 2007/070514, for the treatment of a disorder, such as the disorder described herein. In one embodiment, the JAK inhibitor is 2-fluoro-N-methyl-4-(7-(quinolin-6-ylmethyl)imidazo[1,2-b][1,2,4]triazin-2yl)benzamide (compound A17) or its dihydrochloride or the compound described in PCT Publication No. WO 2007/070514. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with 2-fluoro-K-methyl-4-(7-(quinolin-6-ylmethyl)imidazo[1,2-b][1,2,4]triazine -2-yl)benzamide (Compound A17) or dihydrochloride thereof or a compound described in PCT Publication No. WO 2007/070514 for the treatment of a disorder such as colorectal cancer, myeloid leukemia, hematological malignancy, autoimmune disease, non-Hodgkin's lymphoma or thrombocythemia.

In one embodiment, a JAK inhibitor or 2-fluoro-K-methyl-4-(7-(quinolin-6ylmethyl)umidazo[1,2-b][1,2,4]truαzin-2-yl)benzαmide (Compound A17 ) or its dihydrochloride is administered at a dose of about 400-600 mg (eg per day), for example about 400, 500 or 600 mg or about 400-500 or 500-600 mg.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with the JAK inhibitor, ruxolitinib phosphate (also known as JAKAFI ; Compound A18) or a compound described in PCT Publication No. WO 2007/070514 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the JAK inhibitor is ruxolitinib phosphate (Compound A18) or the compound described in PCT Publication No. WO 2007/070514. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with ruxolitinib phosphate (Compound A18) or a compound described in PCT Publication No. WO 2007/070514 to treat a disorder such as prostate cancer, lymphocytic leukemia, multiple myeloma, lymphoma , lung cancer, leukemia, cachexia, breast cancer, pancreatic cancer, rheumatoid arthritis, psoriasis, colorectal cancer, myeloid leukemia, hematologic malignancy, autoimmune disease, non-Hodgkin's lymphoma, or thrombocythemia.

In one embodiment, the JAK inhibitor or ruxolitinib phosphate (Compound A18) is administered at a dose of about 15-25 mg, eg, twice daily. In some embodiments, the dose is about 15, 20, or 25 mg, or about 15-20 or 20-25 mg.

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a deacetylase (DAC) inhibitor panobinostat (compound A19) or a compound described in PCT Publication No. WO 2014/072493 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the DAC inhibitor is a panobinostat (compound A19) or a compound described in PCT Publication No. WO 2014/072493. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with panobinostat (compound A19), a compound described in PCT Publication No. WO 2014/072493, to treat a disorder such as small cell lung cancer, respiratory tract/chest cancer, prostate cancer, multiple myeloma, myelodysplastic syndrome, bone cancer, non-small cell lung cancer, endocrine malignancy, lymphoma, neurological malignancy, leukemia, HIV/AIDS, immune disorder, transplant rejection, gastric cancer, melanoma, breast cancer, pancreatic cancer cancer of the colon and rectum, glioblastoma multiforme, myeloid leukemia, hematological malignancy, kidney cancer, non-Hodgkin's lymphoma, head and neck cancer, hematopoietic disorders, or liver cancer.

In one embodiment, the DAC inhibitor or panobinostat (compound A19) is administered at a dose of approximately 20 mg (eg, per day).

In another embodiment, an anti-TIM-3 antibody molecule, such as an antibody molecule

- 85 040365 against TIM-3 as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an inhibitor of one or more of cytochrome P450 (e.g. 11B2), aldosterone or angiogenesis, osilodrostat (compound A20 ) or a compound described in PCT Publication No. WO 2007/024945 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the inhibitor of one or more of cytochrome P450 (eg, 11B2), aldosterone, or angiogenesis is osilodrostat (Compound A20) or the compound described in PCT Publication No. WO 2007/024945. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with osilodrostat (Compound A20) or a compound described in PCT Publication No. WO 2007/024945 to treat a disorder such as Cushing's syndrome, hypertension, or to treat heart failure.

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an IAP inhibitor, (S)-N-( (S)-1-cyclohexyl2-((S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl)-2-(methylamino)propanamide (compound A21 ) or a compound described in US 8552003 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the IAP inhibitor is (S)-N-((S)-1-cyclohexyl-2-((S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1- yl)-2-oxoethyl)2-(methylamino)propanamide (Compound A21) or the compound described in US Pat. No. 8,552,003. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with (S)-N-((S) 1-cyclohexyl-2-((S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl)-2-(methylamino)propanamide (compound A21) or a compound described in US Pat. No. 8,552,003 for the treatment of a disorder such as multiple myeloma, breast cancer, ovarian cancer, pancreatic cancer, or a hematopoietic disorder.

In one embodiment, an IAP inhibitor, or (S)-N-((S)-1-cyclohexyl-2-((S)-2-(4-(4fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl) -2-oxoethyl)-2-(methylamino)propanamide (compound A21), or the compound described in US 8552003, is administered at a dose of approximately 1800 mg, for example once a week.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with the Smoothened (SMO) inhibitor, sonidegib phosphate (compound A22), (R)-2-(5-(4-(6-benzyl-4,5-dimethylpyridazin-3-yl)-2-methylpiperazin-1yl)pyrazin-2-yl)propan-2-ol (compound A25) or a compound described in PCT Publication No. WO 2007/131201 or WO 2010/007120 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the SMO inhibitor is sonidegiba phosphate (compound A22), (R)-2-(5-(4-(6-benzyl-4,5-dimethylpyridazin-3-yl)-2-methylpiperazin-1yl)pyrazine -2-yl)propan-2-ol (compound A25) or the compound described in PCT Publication No. WO 2007/131201 or WO 2010/007120. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with sonidegib phosphate (compound A22), (R)-2-(5-(4-(6-benzyl-4,5dimethylpyridazin-3-yl)-2-methylpiperazine -1-yl)pyrazin-2-yl)propan-2-ol (Compound A25) or the compound described in PCT Publication No. WO 2007/131201 or WO 2010/007120 for the treatment of a disorder such as cancer, medulloblastoma, small cell lung cancer, prostate cancer, basal cell carcinoma, pancreatic cancer, or inflammation.

In certain embodiments, sonidegib phosphate (Compound A22) is administered at a dose of about 20 to 500 mg, such as about 40 mg to 400 mg, about 50 mg to 300 mg, or about 100 mg to 200 mg, such as about 50 mg, 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. The dosing regimen may vary from, for example, every two days to once a day, two or three times a day.

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with the Alk inhibitor, ceritinib (also known as ZYKADIA; compound A23) or a compound described in PCT Publication No. WO 2007/131201 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the Alk inhibitor is ceritinib (Compound A23) or the compound described in PCT Publication No. WO 2007/131201. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with ceritinib (compound A23) or a compound described in PCT Publication No. WO 2007/131201 to treat a disorder such as non-small cell lung cancer or solid tumors.

In one embodiment, the Alk inhibitor or ceritinib (compound A23) is administered at a dose of about 750 mg, eg, once daily.

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a JAK and/or CDK4/6 inhibitor 7

- 86 040365 cyclopentyl-N,N-dimethyl-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6carboxamide (compound A24) or a compound described in US Pat. No. 8,415,355 or US Pat. No. 8,685,980 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the JAK and/or CDK4/6 inhibitor is 7-cyclonentyl-N,Ndimethyl-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[2, 3-d]pyrimidine-6-carboxamide (Compound A24) or the compound described in US Pat. No. 8,415,355 or US Pat. No. 8,685,980. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with ((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide (compound A24) or a compound described in US 8415355 or US 8685980, for the treatment of a disorder such as lymphoma, neurological malignancy, melanoma, breast cancer or solid tumor.

In one embodiment, a JAK and/or CDK4/6 inhibitor or 7-cyclopentyl-N,N-dimethyl-2((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[2, 3-d]pyrimidine-6-carboxamide (compound A24) is administered at a dose of about 200-600 mg, eg per day.

In one embodiment, the compound is administered at a dose of about 200, 300, 400, 500, or 600 mg, or about 200-300, 300-400, 400-500, or 500-600 mg.

In another embodiment, an antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a prolactin receptor (PRLR) inhibitor, a human monoclonal antibody molecule (compound A26 ) as described in US Pat. No. 7,867,493 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the PRLR inhibitor is a human monoclonal antibody (compound A26) described in US 7,867,493. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with a human monoclonal antibody (compound A26) as described in US 7,867,493 , for the treatment of a disorder such as cancer, prostate cancer, or breast cancer.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a PIM kinase inhibitor N-(4-( (1R,3S,5S)3-amino-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide (Compound A27) or the compound described in PCT Publication No. WO 2010/ 026124, for the treatment of a disorder, such as the disorder described herein. In one embodiment, the PIM kinase inhibitor is N-(4-((lR,3S,5S)-3-amino-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6difluorophenyl)-5-fluoropicolinamide (compound A27) or the compound described in PCT Publication No. WO 2010/026124. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with N-(4-((1R,3S,5S)-3-amino-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6- difluorophenyl)-5fluoropicolinamide (compound A27) or the compound described in PCT Publication No. WO 2010/026124 for the treatment of a disorder such as multiple myeloma, myelodysplastic syndrome, myeloid leukemia or non-Hodgkin's lymphoma.

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a Wnt 2-(2', 3dimethyl-[2,4'-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2-yl)acetamide (Compound A28) or the compound described in PCT Publication No. WO 2010/ 101849, for the treatment of a disorder, such as the disorder described herein. In one embodiment, the Wnt signaling inhibitor is 2-(2',3-dimethyl-[2,4'-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2yl) acetamide (compound A28) or the compound described in PCT Publication No. WO 2010/101849. In one embodiment, the Wnt signaling inhibitor is 2-(2',3-dimethyl-[2,4'bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2-yl )acetamide (compound A28). In one embodiment, the TIM-3 antibody molecule is used in combination with 2-(2',3-dimethyl-[2,4'-bipyridin]-5yl)-N-(5-(pyrazin-2-yl)pyridin-2 -yl)acetamide (Compound A28) or a compound described in PCT Publication No. WO 2010/101849 for the treatment of a disorder such as a solid tumor (e.g. head and neck cancer, squamous cell carcinoma, breast cancer, pancreatic cancer, or colon cancer). intestines).

In certain embodiments, 2-(2',3-dimethyl-[2,4'-bipyridin]-5-yl)-N-(5-(pyrazin-2yl)pyridin-2-yl)acetamide (Compound A28) is administered at a dose of about 1 to 50 mg, such as about 2 mg to 45 mg, about 3 mg to 40 mg, about 5 mg to 35 mg, 5 mg to 10 mg, or about 10 mg to 30 mg, for example approximately 2 mg, 5 mg, 10 mg, 20 mg, 30 mg, or 40 mg. The dosing regimen may vary from, for example, once every two days to once a day, two or three times a day.

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more

- 87 040365 other immunomodulators are used in combination with the BRAF inhibitor encorafenib (Compound A29) or the compound described in PCT Publication No. WO 2011/025927 to treat a disorder, such as the disorder described herein. In one embodiment, the BRAF inhibitor is encorafenib (Compound A29) or the compound described in PCT Publication No. WO 2011/025927. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with encorafenib (Compound A29) or a compound described in PCT Publication No. WO 2011/025927 to treat a disorder such as non-small cell lung cancer, melanoma, or colorectal cancer.

In one embodiment, the BRAF inhibitor or encorafenib (compound A29) is administered at a dose of about 200-300, 200-400, or 300-400 mg, eg, per day. In one embodiment, the compound is administered at a dose of about 200, about 300, or about 400 mg.

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with the CDK4/6 inhibitor 7-cyclopentyl-K, Kdimethyl-2-((5-((1R,6S)-9-methyl-4-oxo-3,9-diazabicyclo[4.2.1]nonan-3-yl)pyridin-2-yl)amino)- 7Hpyrrolo[2,3-d]pyrimidine-6-carboxamide (Compound A30) or the compound described in PCT Publication No. WO 2011/101409 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the CDK4/6 inhibitor is 7-cyclopentyl-K,Kdimethyl-2-((5-((1R,6S)-9-methyl-4-oxo-3,9-diazabicyclo[4,2,1 ]nonan-3-yl)pyridin-2-yl)amino)-7Hpyrrolo[2,3-d]pyrimidine-6-carboxamide (Compound A30) or the compound described in PCT Publication No. WO 2011/101409. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with 7-cyclopentyl-N,N-dimethyl-2-((5-((1R,6S)-9-methyl-4-oxo-3,9-diazabicyclo [4,2,1]nonan-3-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide (compound A30) or the compound described in PCT Publication No. WO 2011/101409, for the treatment of a disorder such as cancer, mantle cell lymphoma, liposarcoma, non-small cell lung cancer, melanoma, squamous cell carcinoma of the esophagus, or breast cancer.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a HER3 inhibitor, compound A31, or a compound described in PCT Publication No. WO 2012/022814, for the treatment of a disorder, such as the disorder described herein. In one embodiment, the HER3 inhibitor is Compound A31 or the compound described in PCT Publication No. WO 2012/022814. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with Compound A31 or a compound described in PCT Publication No. WO 2012/022814 to treat a disorder such as gastric cancer, esophageal cancer, head and neck cancer, squamous cell carcinoma, cancer stomach, breast cancer (eg, metastatic breast cancer), or gastrointestinal/gastrointestinal cancer.

In some embodiments, Compound A31 is a human monoclonal antibody molecule.

In one embodiment, the HER3 inhibitor or Compound A31 is administered at a dose of about 3, 10, 20, or 40 mg/kg, such as once a week (QW). In one embodiment, the compound is administered at a dose of about 3-10, 10-20, or 20-40 mg/kg.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an FGFR2 and/or FGFR4 inhibitor, Compound A32, or a compound described in PCT Publication No. WO 2014/160160 (e.g., anti-FGFR2 and/or FGFR4 antibody-drug conjugate, e.g., mAb 12425) for the treatment of a disorder, e.g., a disorder described herein. In one embodiment, the FGFR2 and/or FGFR4 inhibitor is an A32 compound or a compound described in PCT Publication No. WO 2014/160160. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with Compound A32 or a compound as described in Table 1. 6 for treating a disorder such as cancer, gastric cancer, breast cancer, rhabdomyosarcoma, liver cancer, adrenal cancer, lung cancer, esophageal cancer, colon cancer, or endometrial cancer.

In some embodiments, Compound A32 is an anti-FGFR2 and/or FGFR4 antibody-drug conjugate, such as mAb 12425.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an M-CSF inhibitor, an A33 compound, or a compound described in PCT Publication No. WO 2004/045532 (eg, an antibody molecule or Fab fragment against M-CSF), for the treatment of a disorder, such as the disorder described herein. In one embodiment, the M-CSF inhibitor is Compound A33 or the compound described in PCT Publication No. WO 2004/045532. In one embodiment, the molecule

Anti-TIM-3 antibodies are used in combination with Compound A33 or a compound as described in PCT Publication No. WO 2004/045532 to treat a disorder such as malignancy, prostate cancer, breast cancer, or pigmented villesonodular synovitis (PVNS ).

In embodiments, compound A33 is an anti-M-CSF monoclonal antibody molecule or a fragment (eg, a Fab fragment) thereof. In embodiments, the M-CSF inhibitor or Compound A33 is administered at an average dose of about 10 mg/kg.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a MEK inhibitor, binimetinib (compound A34) or compound described in PCT Publication No. WO 2003/077914 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the MEK inhibitor is binmetinib (Compound A34) or the compound described in PCT Publication No. WO 2003/077914. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with binmetinib (compound A34) or a compound described in PCT Publication No. WO 2003/077914 to treat a disorder such as non-small cell lung cancer, multisystem genetic disorder, melanoma, ovarian cancer , cancer of the digestive tract/gastrointestinal cancer, rheumatoid arthritis, or cancer of the colon and rectum.

In one embodiment, the MEK inhibitor or binimetinib (compound A34) is administered at a dose of approximately 45 mg, eg, twice daily.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an inhibitor of one or more of the c-KITs, releasing histamine, Flt3 (eg FLK2/STK1) or PKC midostaurin (compound A35) or the compound described in PCT Publication No. WO 2003/037347 for the treatment of a disorder, eg the disorder described herein. In one embodiment, the inhibitor is midostaurin (Compound A35) or a compound described in PCT Publication No. WO 2003/037347. In one embodiment, the inhibitor of one or more of c-KIT, histamine release, Flt3 (eg, FLK2/STK1), or PKC is midostaurin. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with midostaurin (Compound A35) or a compound described in PCT Publication No. WO 2003/037347 to treat a disorder such as cancer, colorectal cancer, myeloid leukemia, myelodysplastic syndrome, age-related macular degeneration, diabetic complication, or dermatological disorder.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a TOR inhibitor (e.g., an mTOR inhibitor) everolimus ( also known as AFINITOR; compound A36) or the compound described in PCT Publication No. WO 2014/08 5318 for the treatment of a disorder, such as the disorder described herein). In one embodiment, the TOR inhibitor is everolimus (compound A36) or the compound described in PCT Publication No. WO 2014/085318. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with everolimus (compound A36) to treat a disorder such as interstitial lung disease, small cell lung cancer, respiratory/chest cancer, prostate cancer, multiple myeloma, sarcoma, age-related macular degeneration, bone cancer, tuberous sclerosis, non-small cell lung cancer, endocrine malignancy, lymphoma, neurological disorders, astrocytoma, cervical cancer, neurological malignancy, leukemia, immune disorders, transplant rejection, gastric cancer, melanoma, epilepsy , breast cancer or bladder cancer.

In one embodiment, the TOR inhibitor or everolimus (compound A36) is administered at a dose of about 2.5-20 mg/day. In one embodiment, the compound is administered at a dose of about 2.5, 5, 10, or 20 mg/day, such as about 2.5-5, 5-10, or 10-20 mg/day.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an inhibitor of one or more of VEGFR-2, PDGFR -beta, KIT or Raf-kinase C 1-methyl-5-((2-(5-(trifluoromethyl)-1H-imidazol-2yl)pyridin-4-yl)oxy)-N-(4-(trifluoromethyl)phenyl )-1H-benzo[d]imidazol-2-amine (compound A37) or the compound described in PCT Publication No. WO 2007/030377 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the inhibitor of one or more of VEGFR-2, PDGFR-beta, KIT, or Raf kinase C is 1-methyl-5-((2-(5(trifluoromethyl)-1H-imidazol-2-yl)pyridine -4-yl)oxy)-N-(4-(trifluoromethyl)phenyl)-1H-benzo[d]imidazol-2-amine (Compound A37) or the compound described in PCT Publication No. WO 2007/030377. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with 1-methyl-5

- 89 040365 ((2-(5-(trifluoromethyl)-III-imidazol-2-yl)pyridin-4-yl)oxy)-Ni(4-(trifluoromethyl)phenyl)-III-benzo[d]imidazol-2 α-amine (Compound A37) or a compound described in PCT Publication No. WO 2007/030377 for the treatment of a disorder such as cancer, melanoma or solid tumor.

In another embodiment, an anti-TIM-3 antibody molecule, such as an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a somatostatin agonist and/or growth hormone releasing inhibitor, pasireotide diaspartate (also known as SIGNIFOR; Compound A38) or the compound described in PCT Publication No. WO 2002/010192 or US Pat. No. 7,473,761 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the somatostatin agonist and/or growth hormone releasing inhibitor is pasireotide diaspartate (Compound A38) or the compound described in PCT Publication No. WO 2002/010192 or US Pat. No. 7,473,761. In one embodiment, an anti-TIM-3 antibody molecule is used. in combination with pasireotide diaspartate (Compound A38) or a compound described in PCT Publication No. WO 2002/010192 or US Pat. No. 7,473,761 for the treatment of a disorder such as prostate cancer, endocrine malignancy, neurological malignancy, skin cancer melanoma), pancreatic cancer, liver cancer, Cushing's syndrome, gastrointestinal disorder, acromegaly, liver and biliary tract disorder, or cirrhosis.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a signal transduction modulator and/or angiogenesis inhibitor, dovitinib (Compound A39) or a compound described in PCT Publication No. WO 2009/115562 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the signal transduction modulator and/or angiogenesis inhibitor is dovitinib (Compound A39) or the compound described in PCT Publication No. WO 2009/115562. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with dovitinib (compound A39) or a compound described in PCT Publication No. WO 2009/115562 to treat a disorder such as cancer, respiratory tract cancer/thoracic cancer , multiple myeloma, prostate cancer, non-small cell lung cancer, endocrine malignancy, or neurological genetic disorder.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an EGFR inhibitor (R,E)-N- (7-Chloro-1-(1-(4(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d]imidazol-2-yl)-2-methylisonicotinamide (compound A40) or compound described in PCT Publication No. WO 2013/184757 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the EGFR inhibitor is (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1Hbenzo[d]imidazole -2-yl)-2-methylisonicotinamide (Compound A40) or the compound described in PCT Publication No. WO 2013/184757. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl) -1H-benzo[d]imidazol-2-yl)-2-methylisonicotinamide (compound A40) or the compound described in PCT Publication No. WO 2013/184757 for the treatment of a disorder such as a malignant tumor, for example a solid tumor.

In one embodiment, an EGFR inhibitor or (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but2-enoyl)azepan-3-yl)-1H-benzo[d]imidazole- 2-yl)-2-methylisonicotinamide (Compound A40) is administered at a dose of 150-250 mg, for example per day. In one embodiment, the compound is administered at a dose of about 150, 200, or 250 mg, or about 150-200 or 200-250 mg.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an ALK inhibitor N ⁶ -(2-isopropoxy- 5methyl-4-(1-methylpiperidin-4-yl)phenyl)-N ⁴ -(2-(isopropylsulfonyl)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-4,6-diamine (compound A42) or a compound described in PCT Publication No. WO 2008/073687 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the ALK inhibitor is N ⁶ -(2-isopropoxy-5-methyl-4-(1-methylpiperidin-4-yl)phenyl)-N ⁴ -(2-(isopropylsulfonyl)phenyl)-1H-pyrazolo[ 3,4-d]pyrimidine-4,6-diamine (Compound A42) or the compound described in PCT Publication No. WO 2008/073687. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with N ⁶ -(2-isopropoxy-5-methyl-4-(1-methylpiperidin-4-yl)phenyl)-N ⁴ -(2-(isopropylsulfonyl)phenyl )-1H-pyrazolo[3,4-d]pyrimidine-4,6-diamine (Compound A42) or the compound described in PCT Publication No. WO 2008/073687 for the treatment of a disorder such as cancer, anaplastic large cell

- 90 040365 fine lymphoma (ALCL), non-small cell lung carcinoma (NSCLC) or neuroblastoma.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an IGF-1R 3-(4-( 4-((5-chloro-4-((5methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)amino)-5-fluoro-2-methylphenyl)piperidin-1-yl)thiethane 1, 1-dioxide (compound A43), 5-chloro-M ² -(2-fluoro-5-methyl-4-(1-(tetrahydro-2H-pyran-4-yl)piperidin-4-yl)phenyl)-N ^4- (5-methyl-1H-pyrazol-3-yl)pyrimidin-2,4-diamine (compound A44) or 5-chloroN2-(4-(1-ethylpiperidin-4-yl)-2-fluoro-5- methylphenyl)-N ⁴ -(5-methyl-1H-pyrazol-3-yl)pyrimidin-2,4-diamine (compound A45) or a compound described in PCT Publication No. WO 2010/002655 for the treatment of a disorder, such as the described disorder . In one embodiment, the IGF-1R inhibitor is 3-(4-(4-((5-chloro-4-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)amino) -5-fluoro2-methylphenyl)piperidin-1-yl)thiethane 1,1-dioxide (compound A43), 5-chloro-N ² -(2-fluoro-5-methyl-4-(1(tetrahydro-2H-pyran -4-yl)piperidin-4-yl)phenyl)-N ⁴ -(5-methyl-1H-pyrazol-3-yl)pyrimidin-2,4-diamine (compound A44), 5-chloro-N2-(4 -(1-ethylpiperidin-4-yl)-2-fluoro-5-methylphenyl)-N ⁴ -(5-methyl-1H-pyrazol-3-yl)pyrimidin-2,4-diamine (compound A45) or a compound, described in PCT Publication No. WO 2010/002655. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with 3-(4-(4-((5-chloro-4-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2- yl)amino)-5-fluoro-2-methylphenyl)piperidin-1-yl)thiethane 1,1-dioxides (compound A43), 5-chloro-N ² -(2-fluoro-5-methyl-4-(1 (tetrahydro-2H-pyran-4-yl)piperidin-4-yl)phenyl)-N ⁴ -(5-methyl-1H-pyrazol-3-yl)pyrimidin-2,4-diamine (compound A44), 5- chloro-N2-(4-(1-ethylpiperidin-4-yl)-2-fluoro-5-methylphenyl)-N ⁴ -(5-methyl-1Hpyrazol-3-yl)pyrimidin-2,4-diamine (compound A45 ) or a compound described in PCT Publication No. WO 2010/002655 for the treatment of a disorder such as cancer or sarcoma.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a P-glycoprotein 1 inhibitor, valspodar (also known as AMDRAY; Compound A46) or a compound described in EP 296122 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the P-glycoprotein 1 inhibitor is valspodar (compound A46) or a compound described in EP296122. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with valspodar (compound A46) or a compound described in EP296122 for the treatment of a disorder such as cancer or a drug-resistant tumour.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with one or more of the VEGFR inhibitor, vatalanib succinate ( compound A47) or a compound described in EP 296122 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the VEGFR inhibitor is vatalanib succinate (compound A47) or a compound described in EP296122. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with vatalanib succinate (compound A47) or a compound described in EP296122 for the treatment of cancer.

In another embodiment, an anti-TIM-3 antibody molecule, for example an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with an IDH inhibitor or a compound described in WO 2014/ 141104, for the treatment of a disorder, such as the disorder described herein. In one embodiment, the IDH inhibitor is a compound described in PCT Publication No. WO 2014/141104. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with a compound described in WO 2014/141104 to treat a disorder such as cancer.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a BCL-ABL inhibitor or a compound described in the publication PCT No. WO 2013/171639, WO 2013/171640, WO 2013/171641 or WO 2013/171642, for the treatment of a disorder, such as the disorder described herein. In one embodiment, the BCL-ABL inhibitor is a compound described in PCT Publication No. WO 2013/171639, WO 2013/171640, WO 2013/171641, or WO 2013/171642. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with a compound described in PCT Publication No. WO 2013/171639, WO 2013/171640, WO 2013/171641, or WO 2013/171642 to treat a disorder such as cancer.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a c-RAF inhibitor or compound, opioid.

- 91 040365 in PCT Publication No. WO 2014/151616, for the treatment of a disorder, such as the disorder described herein. In one embodiment, the c-RAF inhibitor is an A50 compound or a compound described in PCT Publication No. WO 2014/15161 6. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with a compound described in PCT Publication No. WO 2014/ 151616, for the treatment of a disorder such as cancer.

In another embodiment, an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibody molecule as described herein, alone or in combination with one or more other immunomodulators, is used in combination with a competitive ERK1/2 ATP inhibitor or a compound described in International Patent Application No. PCT/US2014/062913, for the treatment of a disorder, such as the disorder described herein. In one embodiment, the competitive ERK1/2 ATP inhibitor is a compound described in International Patent Application No. PCT/US2014/062913. In one embodiment, an anti-TIM-3 antibody molecule is used in combination with Compound A51 or a compound described in International Patent Application No. PCT/US2014/062913 to treat a disorder such as cancer.

In some embodiments, the anti-TIM-3 antibody molecule is administered in combination with one or more of Compound A8, Compound A17, Compound A23, Compound A24, Compound A27, Compound A29, and Compound A33.

In some embodiments, an anti-TIM-3 antibody molecule is administered in combination with an anti-cancer agent having known activity in an immune cell assay, e.g., in one or more of a huMLR assay, a T cell proliferation assay, and a B cell proliferation assay. Exemplary analysis methods are described below. From this analysis, an IC ₅₀ can be calculated for each test agent. In embodiments, the anti-cancer agent has an IC ₅₀ , eg, 0-1 μM, 1-4 μM, or greater than 4 μM, eg, 4-10 μM or 4-20 μM. In embodiments, the second drug is selected from one or more of: Compound A9, Compound A16, Compound A17, Compound A21, Compound A22, Compound A25, Compound A28, Compound A48, and Compound 49.

In some embodiments, Compound A28 (or a related compound to Compound A28) is administered at a dose of about 5-10 or 10-30 mg. In some embodiments, Compound A22 (or a related compound to Compound A22) is administered at a dose of approximately 200 mg. In some embodiments, Compound A17 (or a related compound to Compound A17) is administered at a dose of about 400-600 mg. In some embodiments, Compound A16 (or a related compound to Compound A16) is administered at a dose of about 400-600 mg p/o per day. In some embodiments, Compound A29 (or a related compound to Compound A29) is administered at a dose of about 200-400 or 300-400 mg. In some embodiments, Compound A24 (or a related compound to Compound A24) is administered at a dose of about 200-600 mg. In some embodiments, compound A23 (ceritinib) (or a ceritinib-related compound) is administered at a dose of approximately 750 mg once daily. In some embodiments, Compound A8 (or a related compound to Compound A8) is administered at a dose of about 200-400 or 300-400 mg. In some embodiments, Compound A5 (or a related compound to Compound A5) is administered at a dose of about 100-125 mg. In some embodiments, Compound A6 (or a related compound to Compound A6) is administered at a dose of approximately 100 mg. In some embodiments, Compound A1 (or a related compound to Compound A1) is administered at a dose of about 200-300 or 200-600 mg. In some embodiments, Compound A40 (or a related compound to Compound A40) is administered at a dose of about 150-250 mg. In embodiments, Compound A10 (or a compound related to Compound A10) is administered at a dose of about 400 to 700 mg, eg, administered three times per week, with administration for 2 weeks and one week without administration. In embodiments of the BCR-ABL inhibitor, the inhibitor is administered at a dose of about 20 mg bid-80 mg bid.

Below is an exemplary huMLR assay and B or T cell proliferation assay.

The reaction of mixed human lymphocytes.

The mixed lymphocyte response (MLR) is a functional assay that measures the proliferative response of one individual's lymphocytes (responder) to another individual's lymphocytes (stimulator). For allogeneic MLR, peripheral blood mononuclear cells (PBMCs) from three donors were isolated from buffy coats of unknown HLA type (Kantonspital Blutspendezentrum from Bern and Aarau, Switzerland). Cells were prepared at 2105 in 0.2 ml culture medium containing RPMI 1640 GlutaMAX™ with 10% fetal calf serum (FCS), 100 U penicillin/100 μg streptomycin, 50 μg 2-mercaptoethanol. Individual 2-targeted reactions were performed by mixing PBMCs from two different donors in a 1:1 ratio and cocultivation was performed in triplicate in 96-well flat bottom tissue culture plates for 6 days at 37°C, 5% CO ₂ , in the presence of or in the absence of a range of concentrations of the studied compounds from 8 points. Cells were treated with 3H-TdR (1 mCi/0.2 ml) during the last 16 h of culture and the radioactivity incorporated therein was used as a measure.

- 92 040365 cell proliferation. For each compound, the concentration that inhibited 50% of the maximum huMLR response (IC ₅₀ ) was calculated. Cyclosporine was used as a positive control for huMLR inhibition.

Human B cell proliferation assay.

PBMCs were isolated immediately prior to analysis using a FicollPaque density gradient from human blood and subjected to negative B-cell isolation. B cells were resuspended in culture medium (RPMI 1640, HEPES, 10% FCS, 50 μg/ml gentamicin, 50 μM 2-mercaptoethanol, 1x ITS (insulin, transferrin and sodium selenite), 1x non-essential amino acids) at a concentration of 9104 per well in 96-well flat bottom culture plate. B cell stimulation was performed with a human anti-IgM antibody molecule (30 μg/ml) and IL-4 (75 ng/ml) or with a CD40 ligand (3 μg/ml) and IL-4 (75 ng/ml) in the presence of or in the absence of a range of studied concentrations of 7 points. After culture for 72 hours at 37° C., 10% CO ₂ , cells were treated with 3H-TdR (1 μCi/well) for the last 6 hours of culture. B cells were then harvested and thymidine incorporation was measured using a scintillation counter. An average value was calculated for each duplicated treatment and these data were plotted in XLfit 4 to determine the corresponding IC ₅₀ values.

Human T cell proliferation assay.

PBMCs were isolated immediately prior to analysis using a FicollPaque density gradient from human blood and subjected to negative T cell isolation. T cells were prepared in culture medium (RPMI 1640, HEPES, 10% FCS, 50 μg/ml gentamicin, 50 μM 2-mercaptoethanol, 1x ITS (insulin, transferrin and sodium selenite), 1x non-essential amino acids) at a concentration of 8104 per well in 96-well flat bottom culture plate. T cell stimulation was performed with a human anti-CD3 antibody molecule (10 μg/mL) or with an anti-human CD3 antibody molecule (5 μg/mL) and an anti-CD28 antibody molecule (1 μg/mL) in the presence or absence of a range of test concentrations from 7 points. After culture for 72 hours at 37° C., 10% CO ₂ , cells were treated with 3H-TdR (1 μCi/well) for the last 6 hours of culture. Cell proliferation was measured by thymidine incorporation allowing determination of the IC ₅₀ for each test compound.

Means that suppress the immune system.

In an alternative embodiment, the anti-TIM-3 antibody molecules described herein are used to generate anti-idiotypic peptides or antibodies (Wallmann, J. et al. (2010) Anti-Ids in Allergy: Timeliness of a Classic Concept, World Allergy Organiz. J. 3(6):195-201 Nardi, M. et al.(2000) Antiidiotype Antibody Against Platelet Anti-Gpiiia Contributes To The Regulation Of Thrombocytopenia In HIV-1-ITP Patients, J. Exp. Med. 191( 12):2093-2100) or mimetics (Zang, Y. C. et al. (2003) Human Anti-Idiotypic T Cells Induced By TCR Peptides Corresponding To A Common CDR3Sequence Motif In Myelin Basic Protein-Reactive T Cells, Int. Immunol. 15( 9):1073-1080 Loiarro, M. et al (Epub 2010 Apr. 8) Targeting TLR/IL-1R Signaling In Human Diseases, Mediators Inflamm. 2010:674363) against TIM-3. Such molecules can serve as substitutes for TIM-3, and thus their administration to an individual suppresses the immune system of that individual by mimicking or promoting TIM-3 ligand binding. Such molecules are useful in the treatment of graft versus host disease. Similarly, agonist antibodies that i) enhance binding between such antibodies and such receptor/ligand or ii) trigger signaling when bound directly to a TIM-3 or TIM-3 ligand are useful as TIM-3 signal transduction agonists. and thus are useful in the treatment of inflammation and autoimmune disease through direct or indirect antagonism of receptor activity.

Bispecific antibodies exhibiting immunospecific binding to both TIM-3 and TIM-3 ligands are able to bind to both APCs and T cells and thus promote colocalization of APCs and T cells. Such co-localization facilitates the ability of such cells to communicate with each other through the TIM-3 ligand and TIM-3 molecules that are not complexed with the antibody, or through co-inhibitory molecules. Such binding provides suppression of the immune system of the recipient.

Suppression of the immune system is desirable in the treatment of inflammatory and autoimmune diseases, and graft-versus-host disease (GvHD). Examples of autoimmune disorders that can be treated by administering the antibodies of the present invention include, but are not limited to, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, autoimmune adrenal disease, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune oophoritis, and orchitis, autoimmune thrombocytopenia, Behçet's disease, bullous pemphigoid, cardiomyopathy, abdominal aphthae-dermatitis, chronic fatigue and immune dysfunction syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, Churg-Strauss syndrome, cicatricial pemphigoid, CREST syndrome, cold agglutinin disease, Crohn's disease, discoid lupus, essential mixed cryoglobulinemia, fibromyalgia-fibromyositis, glomerulonephritis, Graves' disease, Guillain-Barré syndrome, ti

- 93 040365 Hashimoto's rheoiditis, idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura (ITP), IgA neuropathy, juvenile arthritis, lichen planus, lupus vulgaris, Meniere's disease, mixed connective tissue disease, multiple sclerosis, neuromyelitis optics (NMO), diabetes mellitus Type 1 or immune-mediated diabetes mellitus, myasthenia gravis, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychondritis, polyglandular syndromes, polymyalgia rheumatica, polymyositis and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud's syndrome, Reiter's syndrome , sarcoidosis, scleroderma, Sjogren's syndrome, stiffness syndrome, systemic lupus erythematosus, lupus vulgaris, Takayasu's arteritis, temporal arteritis/giant cell arteritis, transverse myelitis, ulcerative colitis, uveitis, vasculitis such as dermatitis herpetiformis, vasculitis, vitiligo, and Wegener's granulomatosis.

Examples of inflammatory disorders that can be prevented, treated, or controlled by the methods of the invention include, but are not limited to, asthma, encephalitis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), allergic disorders, septic shock, pulmonary fibrosis, undifferentiated spondyloarthropathy, undifferentiated arthropathy, arthritis, inflammatory osteolysis, and chronic inflammation due to chronic viral or bacterial infections.

Thus, the antibodies and antigen-binding fragments of the present invention can be used to treat inflammatory and autoimmune diseases.

Diagnostic Application

In some aspects, the present invention relates to a diagnostic method for detecting the presence of a TIM-3 protein in vitro (for example, in a biological sample, such as a tissue biopsy, for example, from malignant tissue) or in vivo (for example, imaging in an individual in vivo). The method includes: (i) contacting a sample with an antibody molecule as described herein, or administering an antibody molecule to an individual; (optionally), (ii) contacting a reference sample, such as a control sample (eg, a biological control sample such as plasma, tissue, biopsy) or a control individual with an antibody molecule described herein; and (iii) detecting complex formation between the antibody molecule and the sample or individual or control sample or individual, wherein a change, e.g., a statistically significant change, in complex formation in the sample or individual relative to the control sample or individual indicates the presence of TIM-3 in the sample. The antibody molecule can be directly or indirectly labeled with a detectable substance to facilitate detection of bound or unbound antibody. Suitable detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, and radioactive materials, as described above and as described in more detail below.

The term sample, as used for samples used to detect polypeptides, includes, but is not limited to, cells, cell lysates, extracts of proteins or cell membranes, body fluids such as blood or tissue samples.

Complex formation between an antibody molecule and TIM-3 can be detected by measuring or visualizing either the antibody molecule bound to the TIM-3 antigen or the unbound antibody molecule. Any suitable detection assay can be used, and common detection assays include enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), or tissue immunohistochemistry. As an alternative to labeling an antibody molecule, the presence of TIM-3 in a sample can be analyzed by a competitive immunoassay using detectably labeled standards and an unlabeled antibody molecule. In that assay, a biological sample, labeled standards, and an antibody molecule are combined and the amount of labeled standard bound to an unlabeled binding molecule is determined. The amount of TIM-3 in a sample is inversely proportional to the amount of labeled standard bound to the antibody molecule.

In some aspects, the present invention relates to methods that use an anti-TIM-3 antibody molecule to diagnose sepsis, SIRS (systemic inflammatory response syndrome), preeclampsia, or glomerulonephritis. Sepsis is often accompanied by TIM-3 suppression (Yang et al., J Immunol. 2013 Mar 1;190(5):2068-79), so that decreased TIM-3 levels indicate sepsis, while normal TIM-3 levels are a sign that sepsis is absent. In SIRS and preeclampsia, TIM-3 levels are decreased in peripheral lymphocytes (Miko et al., PLoS ONE 8(8):e71811), such that decreased TIM-3 levels are indicative of SIRS or preeclampsia, while normal TIM-3 levels indicate that SIRS and preeclampsia are not present. In glomerulonephritis, TIM-3 can be activated (see Schroll et al., Am J Pathol 2010 April; 176(4):1716-1742), so that elevated TIM-3 levels indicate glomerulonephritis while normal levels indicate that glomerulonephritis is not present.

Nucleic acids

The present invention also relates to nucleic acids containing nucleotide

- 94 040365 sequences that encode the variable regions of the heavy and light chains and CDRs of anti-TIM-3 antibody molecules as described herein. For example, the present invention relates to first and second nucleic acids encoding the heavy and light chain variable regions, respectively, of an anti-TIM-3 antibody molecule selected from one or more of the antibody molecules described herein, for example, the antibody shown in Table 1. 1-4. The nucleic acid may comprise a nucleotide sequence encoding any of the amino acid sequences shown in the tables herein, or a sequence substantially identical thereto (e.g., a sequence that is at least about 85%, 90%, 95%, 99%, or more identical to it or differing by no more than 3, 6, 15, 30, or 45 nucleotides from the sequences shown in Tables 1-4).

In certain embodiments, the implementation of the nucleic acid may contain a nucleotide sequence encoding at least one, two or three CDRs from the variable region of the heavy chain having the amino acid sequence as listed in table. 1-4, or a sequence substantially homologous to it (eg, a sequence at least about 85%, 90%, 95%, 99% or more identical to it and/or having one or more substitutions, for example, conservative substitutions). In some embodiments, the implementation of the nucleic acid may contain a nucleotide sequence encoding at least one, two or three CDRs from the variable region of the light chain having the amino acid sequence as listed in table. 1-4, or a sequence substantially homologous thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical to it and/or having one or more substitutions, e.g., conservative substitutions) . In some embodiments, the implementation of the nucleic acid may contain a nucleotide sequence encoding at least one, two, three, four, five, or six CDRs from the variable regions of the heavy and light chains having the amino acid sequence as listed in table. 1-4, or a sequence substantially homologous thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical to it and/or having one or more substitutions, e.g., conservative substitutions) .

In certain embodiments, the implementation of the nucleic acid may contain a nucleotide sequence encoding at least one, two or three CDRs from the variable region of the heavy chain having a nucleotide sequence, as listed in table. 1-4, or a sequence substantially homologous to it (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical to it and/or capable of hybridizing under the stringency conditions described herein) . In some embodiments, the implementation of the nucleic acid may contain a nucleic acid sequence encoding at least one, two or three CDRs from the variable region of the light chain, having a nucleotide sequence as indicated in table. 1-4, or a sequence substantially homologous to it (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical to it and/or capable of hybridizing under the stringency conditions described herein) . In certain embodiments, the implementation of the nucleic acid may contain a nucleotide sequence encoding at least one, two, three, four, five, or six CDRs from the variable regions of the heavy and light chains having a nucleotide sequence as listed in table. 1-4, or a sequence substantially homologous to it (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical to it and/or capable of hybridizing under the stringency conditions described herein) . Nucleic acids described in the present description include deoxyribonucleotides or ribonucleotides, or their analogues. The polynucleotide can be either single stranded or double stranded, and if it is single stranded, it can be a coding strand or a non-coding (antisense) strand. The polynucleotide may contain modified nucleotides such as methylated nucleotides and nucleotide analogs. The sequence of nucleotides may be interrupted by non-nucleotide components. The polynucleotide can also be modified after polymerization, for example by conjugation with a labeling component. The nucleic acid may be a recombinant polynucleotide or a genome-derived, cDNA, semi-synthetic or synthetic polynucleotide that either does not occur in nature or is linked to another polynucleotide in a non-natural arrangement.

In some aspects, the application relates to host cells and vectors containing the nucleic acids described in the present description. The nucleic acids may be present in a single vector or in separate vectors within the same host cell or in a separate host cell, as described in more detail below.

Vectors

In addition, the present invention provides for vectors containing nucleotide sequences encoding the antibody molecule described in the present description. In some embodiments, the implementation of the vectors contain nucleotides encoding the antibody molecule described in the present description. In some embodiments, the vectors contain nucleotide

- 95 040365 sequences described in the present description. Vectors include, but are not limited to, a virus, plasmid, cosmid, lambda phage, or yeast artificial chromosome (YAC).

Numerous vector systems can be used. For example, one class of vectors uses DNA elements that are derived from animal viruses, such as bovine papilloma virus, polyoma virus, adenovirus, vaccinia virus, vaculovirus, retroviruses (Payca sarcoma virus, MMTV, or MOMLV), or virus SV40. Another class of vectors uses RNA elements derived from RNA viruses such as the Semliki forest virus, eastern equine encephalitis virus, and flaviviruses.

In addition, the selection of cells that have stably integrated DNA into their chromosomes can be carried out by introducing one or more markers that allow the selection of transfected host cells. The marker may confer, for example, prototropy to an auxotrophic host, resistance to biocides (eg antibiotics), or resistance to heavy metals such as copper or similar. The selectable marker gene can either be directly linked to the DNA sequences to be expressed, or it can be introduced into the same cell by co-transformation. Additional elements may be required for optimal mRNA synthesis. These elements may include splicing signals as well as transcription promoters, enhancers, and termination signals.

Once an expression vector or DNA sequence containing constructs has been obtained for expression, the expression vectors can be transfected or introduced into an appropriate host cell. Various methods can be used to achieve this, such as protoplast fusion, calcium phosphate precipitation, electroporation, retroviral transduction, viral transfection, gene gun, lipid-based transfection, or other conventional methods. In the case of protoplast fusion, cells are grown in medium and screened for appropriate activity.

Methods and conditions for culturing the resulting transfected cells and isolating the produced antibody molecule are known to those skilled in the art and may be varied or optimized depending on the particular expression vector and mammalian host cell used, as described herein.

Cells

The present invention also relates to host cells containing a nucleic acid encoding an antibody molecule as described herein.

In some embodiments, host cells are genetically modified to contain nucleic acids encoding an antibody molecule.

In certain embodiments, host cells are modified by genetic engineering using an expression cassette. Expression cassette refers to nucleotide sequences that are capable of influencing gene expression in hosts compatible with such sequences. Such cassettes may include a promoter, an open reading frame with or without introns, and a termination signal. You can also use additional factors necessary or useful to achieve expression, such as, for example, an inducible promoter.

The invention also relates to host cells containing the vectors described in the present description.

The cell may be, but is not limited to, a eukaryotic cell, a bacterial cell, an insect cell, or a human cell. Suitable eukaryotic cells include, but are not limited to, Vero cells, HeLa cells, COS cells, CHO cells, HEK293 cells, BHK cells, and MDCKII cells. Suitable insect cells include, but are not limited to, Sf9 cells.

Illustrative sequences of antibodies against TIM-3 are described in table. 1-4 below.

- 96 040365

Table 1. Summary of mouse antibody ABTIM3 sequences.

Antibody designation SEQID NO Description ABTIM3 1 Amino acid sequence of VH 2 VL amino acid sequence 3 Amino acid VHCDR1 sequence 4 Amino acid VHCDR2 sequence 5 Amino acid VHCDR3 sequence 6 Amino acid VLCDR1 sequence 7 Amino acid VLCDR2 sequence 8 Amino acid VLCDR3 sequence

Table 2. Amino acid sequence discovery of the heavy chain variable domain and the light chain variable domain of the mouse antibody ABTIM3.

CDRs are shown as white text on a black background.

SEQ ID NO Subsequence 1 QVQLQQPGAE LVKPGASVKM SCKASGYTFT SYNMHWIKQT PGQGLEWIGD IYPGNGDTSY NQKFKGKA;'.'< TADKSSSTVY MQLSSLTSED SAVYYCARVG GAFPMDYWGQ GTSVTVSS 2 DIVLTQSPAS LAVSLGQRAT ISGRASESVE YYGTSLMQWY QQKPGQPPKL L1YAA3NVES GVPARFSGSG SGTDFSLNIH PWEDDIAIY FGQQSRKDPS TFGGGTKLEI K

Table 3. Amino acid sequences of heavy chain CDRs and

Light chain CDR of mouse antibody ABTIM3.

SEQID NO Subsequence 3 SYNMH 4 DIYPGNGDTSYNQKFKG 5 VGGAFPMDY 6 RASESVEYYGTSLMQ 7 AASNVES 8 QQSRKDPST

Illustrative sequences of antibodies against TIM-3 are described in table. 4. Antibody molecules include mouse ABTIM3 and humanized antibody molecules. Shown are the amino acid and nucleotide sequences of the heavy and light chain CDRs, the variable sequences of the heavy and light chains, and the heavy and light chains.

- 97 040365

Table 4 Sequence summary of exemplary anti-TIM-3 antibodies.

Clone hybridoma I I

ABTIM3 I I

SEQ ID NO: 3 (Rabat) ί HCDR1 I SYNMH

SEQ ID NO: 4 (Rabat) I HCDR2 I DIYPGNGDTSYNQRFRG

SEQ ID NO: 5 (Rabat) I HCDR3 I VGGAFPMDY

SEQ ID NO: 9

(Chothia) HCDR1 GYTFTSY SEQ ID NO: 10 (Chothia) SEQ ID NO: 5 (Chothia) SEQ ID NO: 1 SEQ ID NO: 11 SEQ ID NO: 6 (Rabat) SEQ ID NO: 7 (Rabat) SEQ ID NO: 8 (Rabat) SEQ ID NO: 12 (Chothia) SEQ ID NO: 13 (Chothia) SEQ ID NO: 14 (Chothia) SEQ ID NO: 2 SEQ ID NO: 15 ABTIM3-hum01 SEQ ID NO: 3 (Rabat) SEQ ID NO: 4 (Rabat) SEQ ID NO: 5 (Rabat) SEQ ID NO: 9 (Chothia) SEQ ID NO: 10 (Chothia) HCDR2 HDDR3 vh DNA, VH LCDR1 LCDR2 LCDR3 LCDR1 LCDR2 LCDR3 VL DNA, VL HCDR1 HCDR2 HDDR3 HCDR1 HCDR2 YPGNGD VGGAFPMDY QVQLQQPGAELVKPGASVKMSCRASGYTFTSYNMHWIKQTPGQGLEWIGDIYPGNGDTSYNQRF KGKATLTADRSSSTVYMQLSSLTSEDSAVYYCARVGGAFPMDYWGQGTSVTVSS CAGGTGCAACTGCAGCAGCCTGGGGCTGAGCTGGTGAAGCCTGGGGCCTCAGTGAAGATGTCCT GCAAGGCTTCTGGCTACACATTTACCAGTTACAATATGCACTGGATAAAGCAGACACCTGGACA GGGCCTGGAATGGATTGGAGATATTTATCCAGGAAATGGTGATACTTCCTACAATCAGAAATTC AAAGGCAAGGCCACATTGACTGCAGACAAATCCTCCAGCACAGTCTACATGCAGCTCAGCAGCC TGACATCTGAGGACTCTGCGGTCTATTACTGTGCAAGAGTGGGGGGTGCCTTTCCTATGGACTA CTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA RASESVEYYGTSLMQ AASNVES QQSRRDPST SESVEYYGTSL AAS SRRDPS DIVLTQSPASLAVSLGQRATISCRASESVEYYGTSLMQWYQQKPGQPPRLLIYAASNVESGVPA RFSGSGSGTDFSLNIHPVEEDDIAIYFCQQSRKDPSFGGGTKLEIK GACATTGTGCTCACCCAATCTCCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGAGCCACCATCT CCTGCAGAGCCAGTGAAAGTGTTGAATATTATGGCACAAGTTTAATGCAGTGGTACCAACAGAA ACCAGGACAGCCACCCAAACTCCTCATCTATGCTGCATCCAACGTAGAATCTGGGGTCCCTGCC AGGTTTAGTGGCAGTGGGTCTGGGACAGACTTCAGCCTCAACATCCATCCTGTGGAGGAGGATG ATATTGCAATATATTTCTGTCAGCAAAGTAGGAAGGATCCTTCGACGTTCGGTGGAGGCACCAA GCTGGAGATCAAA SYNMH DIYPGNGDTSYNQRFRG VGGAFPMDY GYTFTSY YPGNGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 16 vh QVQLVQSGAEVRRPGASVRVSCRASGYTFTSYNMHWVRQAPGQGLEWIGDIYPGNGDTSYNQRF RGRATMTADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSS SEQ ID NO: 17 DNA, VH CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTGTGAAAGTCTCTT GTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGGCA GGGCCTCGAGTGGATCGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTT AAGGGTAGAGCTACTATGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGC SEQ ID NO: 18 heavy chain QVQLVQSGAEVRRPGASVRVSCRASGYTFTSYNMHWVRQAPGQGLEWIGDIYPGNGDTSYNQRF RGRATMTADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTRGPSVFP LAPCSRSTSESTAALGCLVRDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHRPSNTRVDKRVESRYGPPCPPCPAPEFLGGPSVFLFPPRPKDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNAKTRPREEQFNSTYRWSVLTVLHQDWLNGREYKCR VSNRGLPSSIERTISRARGQPREPQVYTLPPSQEEMTRNQVSLTCLVRGFYPSDIAVEWESNGQ PENNYRTTPPVLDSDGSFFLYSRLTVDRSRWQEGNVFSCSVMHEALHNHYTQRSLSLSLG

- 98 040365

SEQ ID NO: 19 DNA, heavy chain CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTGTGAAAGTCTCTT GTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGGCA GGGCCTCGAGTGGATCGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTT AAGGGTAGAGCTACTATGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCC CTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATT ACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTT CCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCT AGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACA AGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGG TCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAA GTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGG ATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCG CGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAA GTGTCCAACAAGGGACTTCCTAGCT CAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCC GGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATT GACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAG CCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACT CGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCA TGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 (Rabat) LCDR1 RASESVEYYGTSLMQ SEQ ID NO: 7 (Rabat) LCDR2 AASNVES SEQ ID NO: 8 (Rabat) LCDR3 QQSRRDPST SEQ ID NO: 12 (Chothia) LCDR1 SESVEYYGTSL SEQ ID NO: 13 (Chothia) LCDR2 AAS SEQ ID NO: 14 (Chothia) LCDR3 SRRDPS SEQ ID NO: 20 VL DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGGGTRVEIR SEQ ID NO: 21 DNA, VL GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGCGGGCTACTATTA ACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGAT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGG ACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAG SEQ ID NO: 22 Light chain DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYERGLS PRVYACEV SEQ ID NO: 23 DNA, light chain GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGCGGGCTACTATTA ACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGAT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGG ACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAG CTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGG TGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGA CAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAG CATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCA ACAGGGGCGAGTGC ABTIM3-humO2 SEQ ID NO: 3 (Rabat) HCDR1 SYNMH SEQ ID NO: 24 (Rabat) HCDR2 DIYPGSGDTSYNQRFRG SEQ ID NO: 5 (Rabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 25 (Chothia) HCDR2 YPGSGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY

- 99 040365

SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 vh DNA, VH heavy chain QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIYPGSGDTSYNQKF KGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSS CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTGTGAAAGTTAGCT GTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGTCA AGGCCTCGAGTGGATCGGCGATATCTACCCCGGTAGCGGCGACACTAGTTATAATCAGAAGTTT AAGGGTAGAGCTACTATGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAAGATACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGC QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIYPGSGDTSYNQKF KGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTKGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCK VSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 29 SEQ ID NO: 6 (Kabat) SEQ ID NO: 7 (Kabat) SEQ'id NO:8 (Kabat) SEQ ID NO: 12 (Chothia) DNA, heavy chain LCDR1 LCDR2 LCDR3 LCDR1 CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTGTGAAAGTTAGCT GTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGTCA AGGCCTCGAGTGGATCGGCGATATCTACCCCGGTAGCGGCGACACTAGTTATAATCAGAAGTTT AAGGGTAGAGCTACTATGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAAGATACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCC CTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATT ACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTT CCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCT AGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACA AGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGG TCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAA GTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGG ATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCG CGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAA GTGTCCAACAAGGGACTTCCTAGCT CAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCC GGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATT GACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAG CCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACT CGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCA TGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA RASESVEYYGTSLMQ ' AASNVES QQSRKDPST SESVEYYGTSL SEQ ID NO: 13 (Chothia) LCDR2 AAS SEQ ID NO: 14 (Chothia) LCDR3 SRKDPS SEQ ID NO: 20 VL DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGGGTKVEIK SEQ ID NO: 21 DNA, VL GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGCGGGCTACTATTA ACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGAT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGG ACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAG SEQ ID NO: 22 Light chain DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADVEK NGLS FKVYEK PACE

- 100 040365

SEQ ID NO: 23 DNA, light chain GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGCGGGCTACTATTA ACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGAT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGG ACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAG CTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGG TGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGA CAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAG CATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCA ACAGGGGCGAGTGC ABTIM3-humO3 SEQ ID NO: 3 (Kabat) HCDR1 SYNMH SEQ ID NO: 30 (Kabat) HCDR2 DIYPGQGDTSYNQKFKG SEQ ID NO: 5 (Kabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 31 (Chothia) HCDR2 YPGQGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 32 vh QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIYPGQGDTSYNQKF KGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSS SEQ ID NO: 33 DNA, VH CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTGTGAAAGTTAGCT GTAAAGCTAGTGGCTATACTTTCACTTCTTATAATATGCACTGGGTCCGCCAGGCCCCAGGTCA AGGCCTCGAGTGGATCGGCGATATCTACCCCGGTCAAGGCGACACTTCCTATAATCAGAAGTTT AAGGGTAGAGCTACTATGACCGCCGATAAGTCTACTTCTACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCAATGGACTA CTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGC SEQ ID NO: 34 heavy chain QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIYPGQGDTSYNQKF KGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTKGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCK VSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 35 DNA, heavy chain CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTGTGAAAGTTAGCT GTAAAGCTAGTGGCTATACTTTCACTTCTTATAATATGCACTGGGTCCGCCAGGCCCCAGGTCA AGGCCTCGAGTGGATCGGCGATATCTACCCCGGTCAAGGCGACACTTCCTATAATCAGAAGTTT AAGGGTAGAGCTACTATGACCGCCGATAAGTCTACTTCTACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCAATGGACTA CTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCC CTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATT ACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTT CCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCT AGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACA AGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGG TCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAA GTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGG ATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCG CGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAA GTGTCCAACAAGGGACTTCCTAGCT CAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCC GGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATT GACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAG CCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACT CGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCA TGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 (Kabat) LCDR1 RASESVEYYGTSLMQ

- 101 040365

SEQ ID NO: 7 (Rabat) LCDR2 AASNVES SEQ ID NO: 8 (Rabat) LCDR3 QQSRRDPST SEQ ID NO: 12 (Chothia) LCDR1 SESVEYYGTSL SEQ ID NO: 13 (Chothia) LCDR2 AAS SEQ ID NO: 14 (Chothia) LCDR3 SRRDPS SEQ ID NO: 20 VL DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPRLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGGGTKVEIR SEQ ID NO: 21 DNA, VL GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGCGGGCTACTATTA ACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGAT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGG ACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAG SEQ ID NO: 22 Light chain DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPRLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREARVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYER HRVYACEVTHQGLSSPVTRSFNRGEC SEQ ID NO: 23 DNA, light chain GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGCGGGCTACTATTA ACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGAT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGG ACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAG CTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGG TGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGA CAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAG CATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCA ACAGGGGCGAGTGC ABTIM3-humO4 SEQ ID NO: 3 (Rabat) HCDR1 SYNMH SEQ ID NO: 4 (Rabat) HCDR2 DIYPGNGDTSYNQRFRG SEQ ID NO: 5 (Rabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 10 (Chothia) HCDR2 YPGNGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 36 vh QVQLVQSGAEVKRPGASVRVSCKASGYTFTSYNMHWIRQAPGQGLEWIGDIYPGNGDTSYNQKF KGRATLTADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSS SEQ ID NO: 37 DNA, VH CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTGTGAAAGTTTCTT GTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGATTAGACAGGCCCCAGGGCA GGGCCTCGAGTGGATCGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTT AAGGGTAGAGCTACCCTGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGGCAGGGCACCCTGGTCACCGTGTCTAGC SEQ ID NO: 38 heavy chain QVQLVQSGAEVKRPGASVRVSCKASGYTFTSYNMHWIRQAPGQGLEWIGDIYPGNGDTSYNQKF KGRATLTADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTRGPSVFP LAPCSRSTSESTAALGCLVRDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESRYGPPCPPCPAPEFLGGPSVFLFPPKPRDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGREYKCK VSNKGLPSSIERTISKARGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYRTTPPVLDSDGSFFLYSRLTVDRSRWQEGNVFSCSVMHEALHNHYTQRSLSLSLG

- 102040365

SEQ ID NO: 39 DNA, heavy chain CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTGTGAAAGTTTCTT GTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGATTAGACAGGCCCCAGGGCA GGGCCTCGAGTGGATCGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTT AAGGGTAGAGCTACCCTGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGGCAGGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCC CTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATT ACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTT CCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCT AGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACA AGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGG TCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAA GTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGG ATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCG CGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAA GTGTCCAACAAGGGACTTCCTAGCT CAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCC GGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATT GACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAG CCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACT CGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCA TGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 (Rabat) LCDR1 RASESVEYYGTSLMQ SEQ ID NO: 7 (Rabat) LCDR2 AASNVES SEQ ID NO: 8 (Rabat) LCDR3 QQSRRDPST SEQ ID NO: 12 (Chothia) LCDR1 SESVEYYGTSL SEQ ID NO: 13 (Chothia) LCDR2 AAS SEQ ID NO: 14 (Chothia) LCDR3 SRRDPS SEQ ID NO: 40 VL DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYFCQQSRKDPSTFGGGTKVEIR SEQ ID NO: 41 DNA, VL GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGCGGGCTACTATTA ACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGAT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGG ACGTGGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAG SEQ ID NO: 42 Light chain DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYFCQQSRKDPSTTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKECDSTYSLSSTLTLSKADYTHER HRVYACEVRS

- 103 040365

SEQ ID NO: 43 ABTIM3-humO5 SEQ ID NO: 3 (Rabat) SEQ ID NO: 24 (Rabat) SEQ ID NO: 5 (Rabat) SEQ ID NO: 9 (Chothia) SEQID NO: 25(Chothia) SEQ ID NO: 5 (Chothia) SEQ ID NO: 44 SEQ ID NO: 45 SEQ ID NO: 46 DNA, light chain HCDR1 HCDR2 HDDR3 HCDR1 HCDR2 HDDR3 vh DNA, VH heavy chain ^AArATCGTCCKiAC^^ ACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGAT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGG ACGTGGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAG CTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGG TGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGA CAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAG CATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCA ACAGGGGCGAGTGC SYNMH DIYPGSGDTSYNQRFRG VGGAFPMDY GYTFTSY YPGSGD VGGAFPMDY QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWIRQAPGQGLEWIGDIYPGSGDTSYNQRF RGRATLTADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSS CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCAAGCGTTAAAGTCTCAT GTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGATTAGACAGGCCCCAGGGCA AGGCCTGGAGTGGATCGGCGATATCTACCCCGGTAGCGGCGACACTAGTTATAATCAGAAGTTT AAGGGTAGAGCTACCCTGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC TGAGGAGTGAAGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGTCAAGGCACCCTGGTCACCGTGTCAAGC QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWIRQAPGQGLEWIGDIYPGSGDTSYNQRF RGRATLTADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTRGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNARTRPREEQFNSTYRWSVLTVLHQDWLNGREYRCR VSNRGLPSSIERTISRARGQPREPQVYTLPPSQEEMTRNQVSLTCLVRGFYPSDIAVEWESNGQ PENNYRTTPPVLDSDGSFFLYSRLTVDRSRWQEGNVFSCSVMHEALHNHYTQRSLSLSLG SEQ ID NO: 47 DNA, heavy chain CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCAAGCGTTAAAGTCTCAT GTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGATTAGACAGGCCCCAGGGCA AGGCCTGGAGTGGATCGGCGATATCTACCCCGGTAGCGGCGACACTAGTTATAATCAGAAGTTT AAGGGTAGAGCTACCCTGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC TGAGGAGTGAAGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGTCAAGGCACCCTGGTCACCGTGTCAAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCC CTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATT ACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTT CCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCT AGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACA AGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGG TCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAA GTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGG ATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCG CGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAA GTGTCCAACAAGGGACTTCCTAGCT CAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCC GGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATT GACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAG CCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACT CGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCA TGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 (Rabat) LCDR1 RASESVEYYGTSLMQ

- 104 040365

SEQ ID NO: 7 (Rabat) LCDR2 AASNVES SEQ ID NO: 8 (Rabat) LCDR3 QQSRRDPST SEQ ID (Chothia) NO:12 LCDR1 SESVEYYGTSL SEQ ID (Chothia) NO:13 LCDR2 AAS SEQ ID (Chothia) NO:14 LCDR3 SRRDPS SEQ ID NO: 40 VL DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPRLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYFCQQSRKDPSFGGGTKVEIR GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGCGGGCTACTATTA ACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA SEQ ID NO: 41 DNA, VL GCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGAT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGG ACGTGGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAG DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQRPGQPPRLLIYAASNVESGVPD SEQ ID NO: 42 Light chain RFSGSGSGTDFTLTISSLQAEDVAVYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSRADYEK HRVYACEVTHQGLS S PVTRS FNRGEC GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGCGGGCTACTATTA ACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGAT AGGTTTAGCCGGTAGCGCGCTAGTGGCACACTTAGTCACTAG DNA, ACGTGGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA SEQ ID NO: 43 light GGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAG chain CTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGG TGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGA CAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAG CATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCA ACAGGGGCGAGTGGC ABTIM3-humO6 SEQ ID NO: 3 (Rabat) HCDR1 SYNMH SEQ ID NO: thirty (Rabat) HCDR2 DIYPGQGDTSYNQRFRG SEQ ID NO: 5 (Rabat) HDDR3 VGGAFPMDY SEQ ID (Chothia) NO: 9 HCDR1 GYTFTSY SEQ ID (Chothia) NO:31 HCDR2 YPGQGD SEQ ID (Chothia) NO: 5 HDDR3 VGGAFPMDY SEQ ID NO: 48 vh QVQLVQSGAEVKKPGASVRVSCKASGYTFTSYNMHWIRQAPGQGLEWIGDIYPGQGDTSYNQRF RGRATLTADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSS CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTGTGAAAGTCTCTT GTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGATTAGACAGGCCCCAGGTCA SEQ ID NO: 49 DNA, VH AGGCCTCGAGTGGATCGGCGATATCTACCCCGGTCAAGGCGACACTAGTTATAATCAGAAGTTT AAGGGTAGAGCTACCCTGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC QVQLVQSGAEVKKPGASVKVSCRASGYTFTSYNMHWIRQAPGQGLEWIGDIYPGQGDTSYNQRF KGRATLTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTRGPSVFP SEQ ID NO: 50 heavy chain LAPCSRSTSESTAALGCLVRDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPRDTLMISRTPE VTCWVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCR VSNRGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVRGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDRSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

- 105 040365

SEQ ID NO: 51 DNA, heavy chain CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTGTGAAAGTCTCTT GTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGATTAGACAGGCCCCAGGTCA AGGCCTCGAGTGGATCGGCGATATCTACCCCGGTCAAGGCGACACTAGTTATAATCAGAAGTTT AAGGGTAGAGCTACCCTGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCC CTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATT ACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTT CCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCT AGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACA AGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGG TCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAA GTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGG ATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCG CGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAA GTGTCCAACAAGGGACTTCCTAGCT CAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCC GGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATT GACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAG CCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACT CGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCA TGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 (Kabat) LCDR1 RASESVEYYGTSLMQ SEQ ID NO: 7 (Kabat) LCDR2 AASNVES SEQ ID NO: 8 (Kabat) LCDR3 QQSRKDPST SEQ ID NO: 12 (Chothia) LCDR1 SESVEYYGTSL SEQ ID NO: 13 (Chothia) LCDR2 AAS SEQ ID NO: 14 (Chothia) LCDR3 SRKDPS SEQ ID NO: 40 VL DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYFCQQSRKDPSTFGGGTKVEIK SEQ ID NO: 41 DNA, VL GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGCGGGCTACTATTA ACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGAT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGG ACGTGGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAG SEQ ID NO: 42 Light chain DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKS PGLNVYEK

- 106 040365

SEQ ID NO: 43 ABTIM3-humO7 SEQ ID NO: 3 (Rabat) SEQ ID NO: 4 (Rabat) SEQ ID NO: 5 (Rabat) SEQ ID NO: 9 (Chothia) SEQ ID NO: 10 (Chothia) SEQ ID NO: 5 (Chothia) SEQ ID NO: 36 DNA, light chain HCDR1 HCDR2 HDDR3 HCDR1 HCDR2 HDDR3 vh GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGCGGGCTACTATTA ACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGAT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGG ACGTGGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAG CTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGG TGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGA CAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAG CATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCA ACAGGGGCGAGTGC SYNMH DIYPGNGDTSYNQRFRG VGGAFPMDY GYTFTSY YPGNGD VGGAFPMDY QVQLVQSGAEVKKPGASVRVSCRASGYTFTSYNMHWIRQAPGQGLEWIGDIYPGNGDTSYNQKF KGRATLTADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSS SEQ ID NO: 115 DNA, VH Caggtccagctggtccagagcggagcagaggtcaaaaagcccggagcaagcgtgaaggtctcat gcaaagcaagcggatacacatttacatcatacaacatgcactggatcaggcaggctccaggaca gggactggagtggatcggggacatctaccctggaaacggcgatactagctataatcagaagttc aaaggccgggccaccctgacagctgacaagtctactagtaccgtgtatatggagctgagctccc tgcggtctgaagataccgcagtgtactattgcgccagagtcgggggggcatttcctatggatta ttgggggcaggggactctggtcactgtctcctcc SEQ ID NO: 116 SEQ ID NO: 117 heavy chain DNA, heavy chain QVQLVQSGAEVKKPGASVRVSCKASGYTFTSYNMHWIRQAPGQGLEWIGDIYPGNGDTSYNQRF KGRATLTADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTKGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCK VSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVRGFYPSDIAVEWESNGQ PENNYRTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK caggtccagctggtccagagcggagcagaggtcaaaaagcccggagcaagcgtgaaggtctcat gcaaagcaagcggatacacatttacatcatacaacatgcactggatcaggcaggctccaggaca gggactggagtggatcggggacatctaccctggaaacggcgatactagctataatcagaagttc aaaggccgggccaccctgacagctgacaagtctactagtaccgtgtatatggagctgagctccc tgcggtctgaagataccgcagtgtactattgcgccagagtcgggggggcatttcctatggatta ttgggggcaggggactctggtcactgtctcotccgctagcaccaagggcccatccgtcttcoco ctggcgccctgctccaggagcacctccgagagcacagccgccctgggctgcctggtcaaggact acttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacacctt cccggctgtcctacagtcctcaggactctac tccctcagcagcgtggtgaccgtgccctccagc agcttgggcacgaagacctacacctgcaacgtagatcacaagcccagcaacaccaaggtggaca agagagttgagtccaaatatggtcccccatgcccaccatgcccagcacctgagttcctgggggg accatcagtcttcctgttccccccaaaacccaaggacactctcatgatctcccggacccctgag gtcacgtgcgtggtggtggacgtgagccaggaagaccccgaggtccagttcaactggtacgtgg atggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagttcaacagcacgtaccg tgtggtcagcgtcctcaccgtcctgcaccaggactggctgaacggcaaggagtacaagtgcaag gtctccaacaaaggcctcccgtcotcoatcgagaaaaccatctccaaagccaaagggcagcccc gagagccacaggtgtacaccctgcccccatcccaggaggagatgaccaagaaccaggtcagcct gacctgcctggtcaaaggcttctaccccagcgacatcgccgtggagtgggagagcaatgggcag ccggagaacaactacaagaccacgcctcccgtgetggactccgacggctcottettoctetаса gcaggctaaccgtggacaagagcaggtggcaggaggggaatgtcttctcatgctccgtgatgca tgaggctctgcacaaccactacacacagaagagcctctccctgtctctgggtaaa SEQ ID NO: 6 (Rabat) LCDR1 RASESVEYYGTSLMQ

- 107 040365

SEQ ID NO: 7 (Rabat) LCDR2 AASNVES SEQ ID NO: 8 (Rabat) LCDR3 QQSRRDPST SEQ ID NO: 12 (Chothia) LCDR1 SESVEYYGTSL SEQ ID NO: 13 (chothia) LCDR2 AAS SEQ ID NO: 14 (Chothia) LCDR3 SRRDPS SEQ ID NO: 20 VL DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQRPGQPPRLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRRDPSTFGGGTRVEIR SEQ ID NO: 118 DNA, VL Gacatcgtcctgacacagtctcctgacagcctggcagtgagcctgggcgaaagggcaaccatta attgtagagcttccgagtccgtcgagtactatggcactagtctgatgcagtggtaccagcagaa gccagggcagccccctaaactgctgatctatgcagctagcaacgtggagtccggagtcccagac cggttctctggaagtgggtcaggaaccgattttaccctgacaattagctccctgcaggcagaag acgtggccgtctactattgtcagcagagccgcaaggacccaagcacattcggaggggggaccaa agtggaaatcaag SEQ ID NO: 22 Light chain DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQRPGQPPRLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRRDPSTFGGGTRVEIRRTVAAPSVFIFPPSDEQ LRSGTASWCLLNNFYPREARVQWRVDNALQSGNSQESVTEQDSRDSTYSLSSTLEC FRGVTHQGLS PRVYACEVTHQGLS SEQ ID NO: 119 DNA, light chain Gacatcgtcctgacacagtctcctgacagcctggcagtgagcctgggcgaaagggcaaccatta attgtagagcttccgagtccgtcgagtactatggcactagtctgatgcagtggtaccagcagaa gccagggcagccccctaaactgctgatctatgcagctagcaacgtggagtccggagtcccagac cggttctctggaagtgggtcaggaaccgattttaccctgacaattagctccctgcaggcagaag acgtggccgtctactattgtcagcagagccgcaaggacccaagcacattcggaggggggaccaa agtggaaatcaagcggactgttgctgcaccatctgtcttcatcttcccgccatctgatgagcag ttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaag tacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcagga cagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaa cacaaagtctacgcctgcgaagtcacccatcagggcctgagttcaccggtgacaaagagcttca acaggggagagtgt ABTIM3-humO8 SEQ ID NO: 3 (Rabat) HCDR1 SYNMH SEQ ID NO: 4 (Rabat) HCDR2 DIYPGNGDTSYNQRFRG SEQ ID NO: 5 (Rabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 10 (Chothia) HCDR2 YPGNGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 16 vh QVQLVQSGAEVRRPGASVRVSCRASGYTFTSYNMHWVRQAPGQGLEWIGDIYPGNGDTSYNQRF RGRATMTADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSS SEQ ID NO: 120 DNA, VH Caggtccagctggtccagagcggagcagaggtcaaaaagcccggagcaagcgtgaaggtctcat gcaaagcaagcggatacacatttacatcatacaacatgcactgggtcaggcaggctccaggaca gggactggagtggatcggggacatctaccctggaaacggcgatactagctataatcagaagttc aaaggccgggccaccatgacagctgacaagtctactagtaccgtgtatatggagctgagctccc tgcggtctgaagataccgcagtgtactattgcgccagagtcgggggggcatttcctatggatta ttgggggcaggggactctggtcactgtctcctcc SEQ ID NO: 121 heavy chain QVQLVQSGAEVRRPGASVRVSCRASGYTFTSYNMHWVRQAPGQGLEWIGDIYPGNGDTSYNQRF RGRATMTADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTRGPSVFP LAPCSRSTSESTAALGCLVRDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTRTYTCNVDHRPSNTRVDRRVESRYGPPCPPCPAPEFLGGPSVFLFPPRPRDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNARTRPREEQFNSTYRWSVLTVLHQDWLNGREYRCR VSNRGLPSSIERTISRARGQPREPQVYTLPPSQEEMTRNQVSLTCLVRGFYPSDIAVEWESNGQ PENNYRTTPPVLDSDGSFFLYSRLTVDRSRWQEGNVFSCSVMHEALHNHYTQRSLSLSLGR

- 108 040365

SEQ ID NO: 122 DNA, heavy chain caggtccagctggtccagagcggagcagaggtcaaaaagcccggagcaagcgtgaaggtctcat gcaaagcaagcggatacacatttacatcatacaacatgcactgggtcaggcaggctccaggaca gggactggagtggatcggggacatctaccctggaaacggcgatactagctataatcagaagttc aaaggccgggccaccatgacagctgacaagtctactagtaccgtgtatatggagctgagctccc tgcggtctgaagataccgcagtgtactattgcgccagagtcgggggggcatttcctatggatta ttgggggcaggggactctggtcactgtctcotccgctagcaccaagggcccatccgtcttcoco ctggcgccctgctccaggagcacctccgagagcacagccgccctgggctgcctggtcaaggact acttccccgaaccggtgacggtgtcgtggaactcaggcgccctgaccagcggcgtgcacacctt cccggctgtcotacagtcotcaggactctactcootcagcagcgtggtgaccgtgccctccagc agcttgggcacgaagacctacacctgcaacgtagatcacaagcccagcaacaccaaggtggaca agagagttgagtccaaatatggtcccccatgcccaccatgcccagcacctgagttcctgggggg accatcagtcttcctgttccccccaaaacccaaggacactctcatgatctcccggacccctgag gtcacgtgcgtggtggtggacgtgagccaggaagaccccgaggtccagttcaactggtacgtgg atggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagttcaacagcacgtaccg tgtggtcagcgtcctcaccgtcctgcaccaggactggctgaacggcaaggagtacaagtgcaag gtctccaacaaaggcctcccgtcct ccatcgagaaaaccatctccaaagccaaagggcagcccc gagagccacaggtgtacaccctgcccccatcccaggaggagatgaccaagaaccaggtcagcct gacctgcctggtcaaaggcttctaccccagcgacatcgccgtggagtgggagagcaatgggcag ccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctaca gcaggctaaccgtggacaagagcaggtggcaggaggggaatgtcttctcatgctccgtgatgca tgaggctctgcacaaccactacacacagaagagcctctccctgtctctgggtaaa SEQ ID NO: 6 (Rabat) LCDR1 RASESVEYYGTSLMQ SEQ ID NO: 7 (Rabat) LCDR2 AASNVES SEQ ID NO: 8 (Rabat) LCDR3 QQSRRDPST SEQ ID NO: 12 (Chothia) LCDR1 SESVEYYGTSL SEQ ID NO: 13 (Chothia) LCDR2 AAS SEQ ID NO: 14 (Chothia) LCDR3 SRRDPS SEQ ID NO: 40 VL DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQRPGQPPRLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYFCQQSRKDPSTFGGGTKVEIR SEQ ID NO: 123 DNA, VL Gacatcgtcctgacacagtctcctgacagcctggcagtgagcctgggcgaaagggcaaccatta attgtagagcttccgagtccgtcgagtactatggcactagtctgatgcagtggtaccagcagaa gccagggcagccccctaaactgctgatctatgcagctagcaacgtggagtccggagtcccagac cggttctctggaagtgggtcaggaaccgattttaccctgacaattagctccctgcaggcagaag acgtggccgtctacttttgtcagcagagccgcaaggacccaagcacattcggaggggggaccaa agtggaaatcaag SEQ ID NO: 42 Light chain DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQRPGQPPRLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYFCQQSRKDPSFGGGTKVEIRRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLSRADYEK HRVYACEVTHQ

- 109 040365

SEQ ID NO: 124 DNA, light chain Gacatcgtcctgacacagtctcctgacagcctggcagtgagcctgggcgaaagggcaaccatta attgtagagcttccgagtccgtcgagtactatggcactagtctgatgcagtggtaccagcagaa gccagggcagccccctaaactgctgatctatgcagctagcaacgtggagtccggagtcccagac cggttctctggaagtgggtcaggaaccgattttaccctgacaattagctccctgcaggcagaag acgtggccgtctacttttgtcagcagagccgcaaggacccaagcacattcggaggggggaccaa agtggaaatcaagcggactgttgctgcaccatctgtcttcatcttcccgccatctgatgagcag ttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaag tacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcagga cagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaa cacaaagtctacgcctgcgaagtcacccatcagggcctgagttcaccggtgacaaagagcttca acaggggagagtgt ABTIM3-humO9 SEQ ID NO: 3 (Kabat) HCDR1 SYNMH SEQ ID NO: 4 (Kabat) HCDR2 DIYPGNGDTSYNQKFKG SEQ ID NO: 5 (Kabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 10 (Chothia) HCDR2 YPGNGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 52 vh QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGNGDTSYNQKF KGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSS SEQ ID NO: 53 DNA, VH CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCGTGAAAGTTTCTT GTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGGCA AGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTT AAGGGTAGAGTCACTATCACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGTCAAGGCACTACCGTGACCGTGTCTAGC SEQ ID NO: 54 heavy chain QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGNGDTSYNQKF KGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCK VSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

- 110 040365

DNA, CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCGTGAAAGTTTCTT GTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGGCA AGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTT AAGGGTAGAGTCACTATCACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGTCAAGGCACTACCGTGACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCC CTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATT ACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTT CCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCT AGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACA SEQ ID NO: 55 heavy chain AGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGG TCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAA GTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGG ATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCG CGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAA GTGTCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCC GGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATT GACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAG CCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACT CGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCA TGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 (Kabat) LCDR1 RASESVEYYGTSLMQ SEQ ID NO: 7 (Kabat) LCDR2 AASNVES SEQ ID NO: 8 (Kabat) SEQ ID NO: 12 (Chothia) SEQ ID NO: 13 (Chothia) SEQ ID NO: 14 (Chothia) SEQ ID NO: 56 SEQ ID NO: 57 SEQ ID NO: 58 LCDR3 LCDR1 LCDR2 LCDR3 VL DNA, VL light chain QQSRKDPST SESVEYYGTSL AAS SRKDPS EIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLIYAASNVESGIPA RFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSFGGGTKVEIK GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGAGCCCTGGCGAGAGAGCTACACTGA GCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGTCAAGCCCCTAGACTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGGATCCCCGCT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGGAACCCGAGG ATATCGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAAG EIVLTQSPATLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLIYAASNVESGIPA RFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLS PVTEC PVT

- 111 040365

SEQ ID NO: 59 DNA, light chain GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGAGCCCTGGCGAGAGAGCTACACTGA GCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGTCAAGCCCCTAGACTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGGATCCCCGCT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGGAACCCGAGG ATATCGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAG CTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGG TGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGA CAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAG CATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCA ACAGGGGCGAGTGC ABTIM3-humlO SEQ ID NO: 3 (Rabat) HCDR1 SYNMH SEQ ID NO: 4 (Rabat) HCDR2 DIYPGNGDTSYNQRFRG SEQ ID NO: 5 (Rabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 10 (Chothia) HCDR2 YPGNGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 60 vh EVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIYPGNGDTSYNQKF RGQVTISADRSIS TVYLQWS S LRAS DTAMYYCARVGGAFPMDYWGQGT TVTVS S SEQ ID NO: 61 DNA, VH GAAGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAGCCCGGCGAGTCACTGAAGATTAGCT GTAAAGGTTCAGGCTACACCTTCACTAGCTATAATATGCACTGGGTCCGCCAGATGCCCGGGAA AGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTT AAGGGGCAAGTCACAATTAGCGCCGATAAGTCTATTAGCACCGTCTACCTGCAGTGGTCTAGCC TGAAGGCTAGTGACACCGCTATGTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGTCAAGGCACTACCGTGACCGTGTCTAGC SEQ ID NO: 62 heavy chain EVQLVQSGAEVRRPGESLRISCRGSGYTFTSYNMHWVRQMPGRGLEWMGDIYPGNGDTSYNQRF RGQVTISADRSISTVYLQWSSLRASDTAMYYCARVGGAFPMDYWGQGTTVTVSSASTRGPSVFP LAPCSRSTSESTAALGCLVRDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTRTYTCNVDHRPSNTRVDRRVESRYGPPCPPCPAPEFLGGPSVFLFPPRPRDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNARTRPREEQFNSTYRWSVLTVLHQDWLNGREYRCR VSNRGLPSSIERTISRARGQPREPQVYTLPPSQEEMTRNQVSLTCLVRGFYPSDIAVEWESNGQ PENNYRTTPPVLDSDGSFFLYSRLTVDRSRWQEGNVFSCSVMHEALHNHYTQRSLSLSLG

- 112 040365

SEQ ID NO: 63 DNA, heavy chain GAAGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAGCCCGGCGAGTCACTGAAGATTAGCT GTAAAGGTTCAGGCTACACCTTCACTAGCTATAATATGCACTGGGTCCGCCAGATGCCCGGGAA AGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTT AAGGGGCAAGTCACAATTAGCGCCGATAAGTCTATTAGCACCGTCTACCTGCAGTGGTCTAGCC TGAAGGCTAGTGACACCGCTATGTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGTCAAGGCACTACCGTGACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCC CTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATT ACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTT CCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCT AGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACA AGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGG TCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAA GTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGG ATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCG CGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAA GTGTCCAACAAGGGACTTCCTAGCT CAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCC GGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATT GACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAG CCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACT CGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCA TGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 (Rabat) LCDR1 RASESVEYYGTSLMQ SEQ ID NO: 8 (Rabat) LCDR3 QQSRRDPST SEQ ID NO: 12 (Chothia) LCDR1 SESVEYYGTSL SEQ ID NO: 13 (Chothia) LCDR2 AAS SEQ ID NO: 14 (Chothia) LCDR3 SRRDPS SEQ ID NO: 56 VL EIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQRPGQAPRLLIYAASNVESGIPA RFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGGGTRVEIR SEQ ID NO: 57 DNA, VL GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGAGCCCTGGCGAGAGAGCTACACTGA GCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGTCAAGCCCCTAGACTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGGATCCCCGCT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGGAACCCGAGG ATATCGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAG SEQ ID NO: 58 Light chain EIVLTQSPATLSPGERATLSCRASESVEYYGTSLMQWYQQRPGQAPRLLIYAASNVESGIPA RFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGGGTRVEIRRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSVTTYSLSSTLSKADYER HRVYACEVTHQGLS S

- 113 040365

SEQ ID NO: 59 DNA, light chain GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGAGCCCTGGCGAGAGAGCTACACTGA GCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGTCAAGCCCCTAGACTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGGATCCCCGCT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGGAACCCGAGG ATATCGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAG CTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGG TGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGA CAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAG CATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCA ACAGGGGCGAGTGC ABTIM3-humll SEQ ID NO: 3 (Rabat) HCDR1 SYNMH SEQ ID NO: 4 (Rabat) HCDR2 DIYPGNGDTSYNQRFRG SEQ ID NO: 5 (Rabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 10 (Chothia) HCDR2 YPGNGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 52 vh QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGNGDTSYNQKF KGRVTITADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSS SEQ ID NO: 53 DNA, VH CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCGTGAAAGTTTCTT GTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGGCA AGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTT AAGGGTAGAGTCACTATCACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGTCAAGGCACTACCGTGACCGTGTCTAGC SEQ ID NO: 54 heavy chain QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGNGDTSYNQKF KGRVTITADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSSASTRGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPRDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCK VSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVRGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

- 114 040365

SEQ ID NO: 55 DNA, heavy chain CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCGTGAAAGTTTCTT GTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGGCA AGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTT AAGGGTAGAGTCACTATCACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCC TGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGTCAAGGCACTACCGTGACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCC CTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATT ACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTT CCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCT AGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACA AGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGG TCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAA GTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGG ATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCG CGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAA GTGTCCAACAAGGGACTTCCTAGCT CAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCC GGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATT GACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAG CCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACT CGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCA TGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 (Rabat) LCDR1 RASESVEYYGTSLMQ SEQ ID NO: 7 (Rabat) SEQ ID NO: 8 (Rabat) SEQ ID NO: 12 (Chothia) SEQ ID NO: 13 (Chothia) SEQ ID NO: 14 (Chothia) SEQ ID NO: 64 SEQ ID NO: 65 SEQ ID NO: 66 LCDR2 LCDR3 LCDR1 LCDR2 LCDR3 VL DNA, VL light chain AASNVES QQSRRDPST SESVEYYGTSL AAS SRRDPS AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQRPGRAPRLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQPEDFATYFCQQSRRDPSTFGGGTRVEIR GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATAGAGTGACTATCA CCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGGAAAGCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCTCT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGG ACTTCGCTACCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAAG AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQRPGRAPRLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQPEDFATYFCQQSRRDPSTFGGGTRVEIRRTVAAPSVFIFPPSDEQ LRSGTASWCLLNNFYPREARVQWRVDNALQSGNSQESVTEQDSRDSTYSLSSTLTLSRADYER HRVYACEVTHQGLS S P

- 115 040365

SEQ ID NO: 67 DNA, light chain GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATAGAGTGACTATCA CCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGGAAAGCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCTCT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGG ACTTCGCTACCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAG CTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGG TGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGA CAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAG CATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCA ACAGGGGCGAGTGC ABTIM3-huml2 SEQ ID NO: 3 (Kabat) HCDR1 SYNMH SEQ ID NO: 4 (Kabat) HCDR2 DIYPGNGDTSYNQKFKG SEQ ID NO: 5 (Kabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 10 (Chothia) HCDR2 YPGNGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 60 vh EVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIYPGNGDTSYNQKF KGQVTISADKSIS TVYLQWS S LKAS DTAMYYCARVGGAFPMDYWGQGTTVTVSS SEQ ID NO: 61 DNA, VH GAAGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAGCCCGGCGAGTCACTGAAGATTAGCT GTAAAGGTTCAGGCTACACCTTCACTAGCTATAATATGCACTGGGTCCGCCAGATGCCCGGGAA AGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTT AAGGGGCAAGTCACAATTAGCGCCGATAAGTCTATTAGCACCGTCTACCTGCAGTGGTCTAGCC TGAAGGCTAGTGACACCGCTATGTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGTCAAGGCACTACCGTGACCGTGTCTAGC SEQ ID NO: 62 heavy chain EVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIYPGNGDTSYNQKF KGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCK VSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG

- 116 040365

SEQ ID NO: 63 DNA, heavy chain GAAGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAGCCCGGCGAGTCACTGAAGATTAGCT GTAAAGGTTCAGGCTACACCTTCACTAGCTATAATATGCACTGGGTCCGCCAGATGCCCGGGAA AGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTT AAGGGGCAAGTCACAATTAGCGCCGATAAGTCTATTAGCACCGTCTACCTGCAGTGGTCTAGCC TGAAGGCTAGTGACACCGCTATGTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTA CTGGGGTCAAGGCACTACCGTGACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCC CTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATT ACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTT CCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCT AGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACA AGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGG TCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAA GTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGG ATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCG CGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAA GTGTCCAACAAGGGACTTCCTAGCT CAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCC GGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATT GACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAG CCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACT CGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCA TGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 (Rabat) LCDR1 RASESVEYYGTSLMQ SEQ ID NO: 7 (Rabat) SEQ ID NO: 8 (Rabat) SEQ ID NO: 12 (Chothia) SEQ ID NO: 13 (Chothia) SEQ ID NO: 14 (Chothia) SEQ ID NO: 64 SEQ ID NO: 65 SEQ ID NO: 66 LCDR2 LCDR3 LCDR1 LCDR2 LCDR3 VL DNA, VL light chain AASNVES QQSRRDPST SESVEYYGTSL AAS SRRDPS AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPRLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGGGTKVEIR GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATAGAGTGACTATCA CCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGGAAAGCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCTCT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGG ACTTCGCTACCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAAG AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPRLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLEC FRGNSVTHQGL SRVYACEVTHQGL

- 117 040365

SEQ ID NO: 67 DNA, light chain GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATAGAGTGACTATCA CCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGGAAAGCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCTCT AGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGG ACTTCGCTACCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAA GGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAG CTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGG TGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGA CAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAG CATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCA ACAGGGGCGAGTGC ABTIM3-huml3 SEQ ID NO: 3 (Rabat) HCDR1 SYNMH SEQ ID NO: 24 (Rabat) HCDR2 DIYPGSGDTSYNQRFRG SEQ ID NO: 5 (Rabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 25 (Chothia) HCDR2 YPGSGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 68 vh QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGSGDTSYNQKF KGRVTITADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSS SEQ ID NO: 69 DNA, VH CAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCGTCAAGGTGTCCT GCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCACTGGGTCCGACAAGCCCCTGGGCA GGGCTTGGAGTGGATGGGAGACATTTACCCCGGCAGTGGTGACACTTCCTATAACCAGAAGTTC AAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTC TGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTA TTGGGGACAGGGCACAACCGTGACCGTCAGCTCA SEQ ID NO: 70 heavy chain QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGSGDTSYNQKF KGRVTITADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSSASTRGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPRDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCK VSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVRGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

- 118 040365

SEQ ID NO: 71 DNA, heavy chain CAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCGTCAAGGTGTCCT GCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCACTGGGTCCGACAAGCCCCTGGGCA GGGCTTGGAGTGGATGGGAGACATTTACCCCGGCAGTGGTGACACTTCCTATAACCAGAAGTTC AAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTC TGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTA TTGGGGACAGGGCACAACCGTGACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCA CTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACT ACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTT CCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGC TCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATA AAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGG ACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAA GTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGG ACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAG GGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAG GTGTCCAACAAAGGACTGCCAAGCT CTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTA GAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTT GACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAG CCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATT CCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCA CGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 (Rabat) LCDR1 RASESVEYYGTSLMQ SEQ ID NO: 7 (Rabat) LCDR2 AASNVES SEQ ID NO: 8 (Rabat) LCDR3 QQSRRDPST SEQ ID NO: 12 (Chothia) LCDR1 SESVEYYGTSL SEQ ID NO: 13 (Chothia) LCDR2 AAS SEQ ID NO: 14 (Chothia) LCDR3 SRRDPS SEQ ID NO: 64 VL AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQRPGRAPRLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQPEDFATYFCQQSRRDPSTFGGGTRVEIR SEQ ID NO: 125 DNA, VL GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACCGAGTGACAATCA CCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAGCCTGATGCAGTGGTATCAGCAAAA GCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCA AGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCCTTCAGCCAGAGG ACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAA AGTCGAAAATTAAA SEQ ID NO: 66 Light chain AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQRPGRAPRLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQPEDFATYFCQQSRRDPSTFGGGTRVEIRRTVAAPSVFIFPPSDEQ LRSGTASWCLLNNFYPREARVQWRVDNALQSGNSQESVTEQDSRDSTYSLSSTLTLSRADYER HRVYACEVTHQGLS S P

- 119 040365

SEQ ID NO: 126 DNA, light chain GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACCGAGTGACAATCA CCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAGCCTGATGCAGTGGTATCAGCAAAA GCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCA AGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGG ACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAA AGTCGAAATTAAACGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAA CTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGG TCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGA CTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAG CACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCA ACAGAGGAGAATGT ABTIM3-huml4 SEQ ID NO: 3 (Rabat) HCDR1 SYNMH SEQ ID NO: 30 (Rabat) HCDR2 DIYPGQGDTSYNQRFRG SEQ ID NO: 5 (Rabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 31 (Chothia) HCDR2 YPGQGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 72 vh QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGQGDTSYNQKF KGRVTITADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSS SEQ ID NO: 73 DNA, VH CAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCGTCAAGGTGTCCT GCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCACTGGGTCCGACAAGCCCCTGGGCA GGGCTTGGAGTGGATGGGAGACATTTACCCCGGCCAGGGTGACACTTCCTATAACCAGAAGTTC AAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTC TGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTA TTGGGGACAGGGCACAACCGTGACCGTCAGCTCA SEQ ID NO: 74 heavy chain QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGQGDTSYNQKF KGRVTITADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSSASTRGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPRDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCK VSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVRGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

- 120 040365

SEQ ID NO: 75 DNA, heavy chain CAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCGTCAAGGTGTCCT GCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCACTGGGTCCGACAAGCCCCTGGGCA GGGCTTGGAGTGGATGGGAGACATTTACCCCGGCCAGGGTGACACTTCCTATAACCAGAAGTTC AAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTC TGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTA TTGGGGACAGGGCACAACCGTGACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCA CTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACT ACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTT CCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGC TCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATA AAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGG ACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAA GTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGG ACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAG GGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAG GTGTCCAACAAAGGACTGCCAAGCT CTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTA GAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTT GACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAG CCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATT CCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCA CGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 (Kabat) LCDR1 RASESVEYYGTSLMQ SEQ ID NO: 7 (Kabat) SEQ ID NO: 8 (Kabat) SEQ ID NO: 12 (Chothia) SEQ ID NO: 13 (Chothia) SEQ ID NO: 14 (Chothia) SEQ ID NO: 64 SEQ ID NO: 125 SEQ ID NO: 66 LCDR2 LCDR3 LCDR1 LCDR2 LCDR3 VL DNA, VL light chain AASNVES QQSRKDPST SESVEYYGTSL AAS SRKDPS AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSFGGGTKVEIK GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACCGAGTGACAATCA CCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAGCCTGATGCAGTGGTATCAGCAAAA GCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCA AGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGG ACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAA AGTCGAAATTAAA AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKECECTYSLSSTLTLSKADYEKS HKGLVYACEV

- 121 040365

SEQ ID NO: 126 DNA, light chain GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACCGAGTGACAATCA CCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAGCCTGATGCAGTGGTATCAGCAAAA GCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCA AGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGG ACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAA AGTCGAAATTAAACGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAA CTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGG TCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGA CTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAG CACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCA ACAGAGGAGAATGT ABTIM3-huml5 SEQ ID NO: 3 (Kabat) HCDR1 SYNMH SEQ ID NO: 24 (Kabat) HCDR2 DIYPGSGDTSYNQKFKG SEQ ID NO: 5 (Kabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 25 (Chothia) HCDR2 YPGSGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 76 vh EVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIYPGSGDTSYNQKF KGQVTISADKSIS TVYLQWS S LKAS DTAMYYCARVGGAFPMDYWGQGTTVTVSS SEQ ID NO: 77 DNA, VH GAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTTTGAAAATTTCCT GCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCACTGGGTGAGACAGATGCCAGGCAA GGGCCTGGAGTGGATGGGAGACATATACCCAGGCAGTGGAGACACAAGCTATAATCAGAAATTC AAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCAC TTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTA TTGGGGCCAAGGGACCACAGTAACCGTCAGCTCA SEQ ID NO: 78 heavy chain EVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIYPGSGDTSYNQKF KGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCK VSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

- 122 040365

SEQ ID NO: 79 DNA, heavy chain GAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTTTGAAAATTTCCT GCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCACTGGGTGAGACAGATGCCAGGCAA GGGCCTGGAGTGGATGGGAGACATATACCCAGGCAGTGGAGACACAAGCTATAATCAGAAATTC AAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCAC TTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTA TTGGGGCCAAGGGACCACAGTAACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCA CTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACT ACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTT CCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGC TCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATA AAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGG ACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAA GTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGG ACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAG GGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAG GTGTCCAACAAAGGACTGCCAAGCT CTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTA GAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTT GACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAG CCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATT CCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCA CGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 (Rabat) LCDR1 RASESVEYYGTSLMQ !a41LA^ SEQ ID NO: 8 (Rabat) LCDR3 QQSRRDPST SEQ ID NO: 12 (Chothia) LCDR1 SESVEYYGTSL SEQ ID NO: 13 (Chothia) LCDR2 AAS SEQ ID NO: 14 (Chothia) LCDR3 SRRDPS SEQ ID NO: 56 VL EIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLIYAASNVESGIPA RFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGGGTKVEIR SEQ ID NO: 127 DNA, VL GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGCGCGCCACCCTGA GCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATCCCTGATTGCAGTGGTATCAGCAGAA ACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCC AGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGG ACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGGTGGAACCAA GGTAGAGATAAAG SEQ ID NO: 58 Light chain EIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLIYAASNVESGIPA RFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSVTTYSLSSTLSKADYER HRVYACEVTHQGLS S

- 123 040365

SEQ ID NO: 128 DNA, light chain GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGCGCGCCACCCTGA GCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATCCCTGATGCAGTGGTATCAGCAGAA ACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCC AGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGG ACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGGTGGAACCAA GGTAGAGATAAAGCGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAA CTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGG TCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGA CTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAG CACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCA ACAGAGGAGAATGT ABTIM3-huml6 SEQ ID NO: 3 (Rabat) HCDR1 SYNMH SEQ ID NO: 30 (Rabat) HCDR2 DIYPGQGDTSYNQRFRG SEQ ID NO: 5 (Rabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 31 (Chothia) HCDR2 YPGQGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 80 vh EVQLVQSGAEVKRPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIYPGQGDTSYNQRF RGQVTISADRSIS TVYLQWS S LRAS DTAMYYCARVGGAFPMDYWGQGT TVTVS S SEQ ID NO: 81 DNA, VH GAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTTTGAAAATTTCCT GCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCACTGGGTGAGACAGATGCCAGGCAA GGGCCTGGAGTGGATGGGAGACATATACCCAGGCCAGGGAGACACAAGCTATAATCAGAAATTC AAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCAC TTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTA TTGGGGCCAAGGGACCACAGTAACCGTCAGCTCA SEQ ID NO: 82 heavy chain EVQLVQSGAEVRRPGESLRISCRGSGYTFTSYNMHWVRQMPGRGLEWMGDIYPGQGDTSYNQRF RGQVTISADRSISTVYLQWSSLRASDTAMYYCARVGGAFPMDYWGQGTTVTVSSASTRGPSVFP LAPCSRSTSESTAALGCLVRDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTRTYTCNVDHRPSNTRVDRRVESRYGPPCPPCPAPEFLGGPSVFLFPPRPRDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNARTRPREEQFNSTYRWSVLTVLHQDWLNGREYRCR VSNRGLPSSIERTISRARGQPREPQVYTLPPSQEEMTRNQVSLTCLVRGFYPSDIAVEWESNGQ PENNYRTTPPVLDSDGSFFLYSRLTVDRSRWQEGNVFSCSVMHEALHNHYTQRSLSLSLGR

- 124 040365

DNA, GAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTTTGAAAATTTCCT GCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCACTGGGTGAGACAGATGCCAGGCAA GGGCCTGGAGTGGATGGGAGACATATACCCAGGCCAGGGAGACACAAGCTATAATCAGAAATTC AAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCAC TTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTA TTGGGGCCAAGGGACCACAGTAACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCA CTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACT ACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTT CCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGC TCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATA SEQ ID NO: 83 heavy chain AAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGG ACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAA GTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGG ACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAG GGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAG GTGTCCAACAAAGGACTGCCAAGCTCTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTA GAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTT GACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAG CCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATT CCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCA CGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 (Rabat) LCDR1 RASESVEYYGTSLMQ ^AASNVES SEQ ID NO: 8 (Rabat) SEQ ID NO: 12 (Chothia) SEQ ID NO: 13 (Chothia) SEQ ID NO: 14 (Chothia) LCDR3 LCDR1 LCDR2 LCDR3 QQSRRDPST SESVEYYGTSL AAS SRRDPS EIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQRPGQAPRLLIYAASNVESGIPA SEQ ID NO: 56 VL RFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRRDPSTFGGGTRVEIR GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGCGCGCCACCCTGA GCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATCCCTGATTGCAGTGGTATCAGCAGAA ACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCC SEQ ID NO: 127 SEQ ID NO: 58 DNA, VL light chain AGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGG ACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGTGGAACCAA GGTAGAGATAAAG EIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQRPGQAPRLLIYAASNVESGIPA RFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRRDPSTFGGGTRVEIRRTVAAPSVFIFPPSDEQ LRSGTASWCLLNNFYPREARVQWRVDNALQSGNSQESVTEQDSRDSTYSLSSTLTLSRADYER HRVYACEVTHQGLS S PVTRSEC

- 125 040365

SEQ ID NO: 128 DNA, light chain GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGCGCGCCACCCTGA GCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATCCCTGATGCAGTGGTATCAGCAGAA ACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCC AGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGG ACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGGTGGAACCAA GGTAGAGATAAAGCGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAA CTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGG TCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGA CTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAG CACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCA ACAGAGGAGAATGT ABTIM3-huml7 SEQ ID NO: 3 (Rabat) HCDR1 SYNMH SEQ ID NO: 24 (Rabat) HCDR2 DIYPGSGDTSYNQRFRG SEQ ID NO: 5 (Rabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 25 (Chothia) HCDR2 YPGSGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 68 vh QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGSGDTSYNQKF KGRVTITADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSS SEQ ID NO: 69 DNA, VH CAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCGTCAAGGTGTCCT GCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCACTGGGTCCGACAAGCCCCTGGGCA GGGCTTGGAGTGGATGGGAGACATTTACCCCGGCAGTGGTGACACTTCCTATAACCAGAAGTTC AAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTC TGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTA TTGGGGACAGGGCACAACCGTGACCGTCAGCTCA SEQ ID NO: 70 heavy chain QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGSGDTSYNQKF KGRVTITADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSSASTRGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPRDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCK VSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVRGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

- 126 040365

SEQ ID NO: 71 DNA, heavy chain CAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCGTCAAGGTGTCCT GCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCACTGGGTCCGACAAGCCCCTGGGCA GGGCTTGGAGTGGATGGGAGACATTTACCCCGGCAGTGGTGACACTTCCTATAACCAGAAGTTC AAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTC TGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTA TTGGGGACAGGGCACAACCGTGACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCA CTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACT ACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTT CCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGC TCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATA AAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGG ACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAA GTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGG ACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAG GGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAG GTGTCCAACAAAGGACTGCCAAGCT CTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTA GAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTT GACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAG CCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATT CCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCA CGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 (Kabat) LCDR1 RASESVEYYGTSLMQ SEQ ID NO: 7 (Kabat) SEQ ID NO: 8 (Kabat) SEQ ID NO: 12 (Chothia) SEQ ID NO: 13 (Chothia) SEQ ID NO: 14 (Chothia) SEQ ID NO: 56 SEQ ID NO: 127 SEQ ID NO: 58 LCDR2 LCDR3 LCDR1 LCDR2 LCDR3 VL DNA, VL light chain AASNVES QQSRKDPST SESVEYYGTSL AAS SRKDPS EIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLIYAASNVESGIPA RFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSFGGGTKVEIK GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGCGCGCCACCCTGA GCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATCCCTGATGCAGTGGTATCAGCAGAA ACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCC AGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGG ACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGGTGGAACCAA GGTAGAGATAAAG EIVLTQSPATLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLIYAASNVESGIPA RFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLS PVTEC PVT

- 127 040365

SEQ ID NO: 128 DNA, light chain GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGCGCGCCACCCTGA GCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATCCCTGATGCAGTGGTATCAGCAGAA ACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCC AGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGG ACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGGTGGAACCAA GGTAGAGATAAAGCGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAA CTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGG TCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGA CTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAG CACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCA ACAGAGGAGAATGT ABTIM3-huml8 SEQ ID NO: 3 (Rabat) HCDR1 SYNMH SEQ ID NO: 30 (Rabat) HCDR2 DIYPGQGDTSYNQRFRG SEQ ID NO: 5 (Rabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 31 (Chothia) HCDR2 YPGQGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 72 vh QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGQGDTSYNQKF KGRVTITADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSS SEQ ID NO: 73 DNA, VH CAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCGTCAAGGTGTCCT GCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCACTGGGTCCGACAAGCCCCTGGGCA GGGCTTGGAGTGGATGGGAGACATTTACCCCGGCCAGGGTGACACTTCCTATAACCAGAAGTTC AAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTC TGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTA TTGGGGACAGGGCACAACCGTGACCGTCAGCTCA SEQ ID NO: 74 heavy chain QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGQGDTSYNQKF KGRVTITADRSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSSASTRGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPRDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCK VSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVRGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

- 128 040365

SEQ ID NO: 75 DNA, heavy chain CAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCGTCAAGGTGTCCT GCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCACTGGGTCCGACAAGCCCCTGGGCA GGGCTTGGAGTGGATGGGAGACATTTACCCCGGCCAGGGTGACACTTCCTATAACCAGAAGTTC AAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTC TGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTA TTGGGGACAGGGCACAACCGTGACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCA CTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACT ACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTT CCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGC TCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATA AAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGG ACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAA GTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGG ACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAG GGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAG GTGTCCAACAAAGGACTGCCAAGCT CTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTA GAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTT GACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAG CCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATT CCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCA CGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 (Rabat) SEQ ID NO: 8 (Rabat) SEQ ID NO: 12 (Chothia) SEQ ID NO: 13 (Chothia) SEQ ID NO: 14 (Chothia) SEQ ID NO: 56 SEQ ID NO: 127 SEQ ID NO: 58 LCDR1 LCDR3 LCDR1 LCDR2 LCDR3 VL DNA, VL light chain RASESVEYYGTSLMQ ^AASNVES QQSRRDPST SESVEYYGTSL AAS SRRDPS EIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQRPGQAPRLLIYAASNVESGIPA RFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRRDPSTFGGGTRVEIR GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGCGCGCCACCCTGA GCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATCCCTGATGCAGTGGTATCAGCAGAA ACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCC AGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGG ACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGGTGGAACCAA GGTAGAGATAAAG EIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQRPGQAPRLLIYAASNVESGIPA RFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRRDPSTFGGGTRVEIRRTVAAPSVFIFPPSDEQ LRSGTASWCLLNNFYPREARVQWRVDNALQSGNSQESVTEQDSRDSTYSLSSTLTLSRADYER HRVYACEVTHQGLS S PVTRSEC

- 129 040365

SEQ ID NO: 128 DNA, light chain GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGCGCGCCACCCTGA GCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATCCCTGATGCAGTGGTATCAGCAGAA ACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCC AGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGG ACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGGTGGAACCAA GGTAGAGATAAAGCGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAA CTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGG TCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGA CTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAG CACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCA ACAGAGGAGAATGT ABTIM3-huml9 SEQ ID NO: 3 (Kabat) HCDR1 SYNMH SEQ ID NO: 24 (Kabat) HCDR2 DIYPGSGDTSYNQKFKG SEQ ID NO: 5 (Kabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 25 (Chothia) HCDR2 YPGSGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 76 vh EVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIYPGSGDTSYNQKF KGQVTISADKSIS TVYLQWS S LKAS DTAMYYCARVGGAFPMDYWGQGTTVTVSS SEQ ID NO: 77 DNA, VH GAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTTTGAAAATTTCCT GCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCACTGGGTGAGACAGATGCCAGGCAA GGGCCTGGAGTGGATGGGAGACATATACCCAGGCAGTGGAGACACAAGCTATAATCAGAAATTC AAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCAC TTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTA TTGGGGCCAAGGGACCACAGTAACCGTCAGCTCA SEQ ID NO: 78 heavy chain EVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIYPGSGDTSYNQKF KGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCK VSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

- 130 040365

DNA, GAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTTTGAAAATTTCCT GCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCACTGGGTGAGACAGATGCCAGGCAA GGGCCTGGAGTGGATGGGAGACATATACCCAGGCAGTGGAGACACAAGCTATAATCAGAAATTC AAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCAC TTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTA TTGGGGCCAAGGGACCACAGTAACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCA CTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACT ACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTT CCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGC TCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATA SEQ ID NO: 79 heavy chain AAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGG ACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAA GTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGG ACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAG GGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAG GTGTCCAACAAAGGACTGCCAAGCTCTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTA GAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTT GACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAG CCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATT CCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCA CGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 (Rabat) LCDR1 RASESVEYYGTSLMQ SEQ ID NO: 7 (Rabat) LCDR2 AASNVES SEQ ID NO: 8 (Rabat) SEQ ID NO: 12 (Chothia) SEQ ID NO: 13 (Chothia) SEQ ID NO: 14 (Chothia) SEQ ID NO: 64 LCDR3 LCDR1 LCDR2 LCDR3 VL QQSRRDPST SESVEYYGTSL AAS SRRDPS AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQRPGRAPRLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQPEDFATYFCQQSRRDPSTFGGGTRVEIR GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACCGAGTGACAATCA CCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAGCCTGATGCAGTGGTATCAGCAAAA GCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCA SEQ ID NO: 125 SEQ ID NO: 66 DNA, VL light chain AGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTTCAGCCAGAGG ACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGTGGGACAAA AGTCGAAATTAAA AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQRPGRAPRLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQPEDFATYFCQQSRRDPSTFGGGTRVEIRRTVAAPSVFIFPPSDEQ LRSGTASWCLLNNFYPREARVQWRVDNALQSGNSQESVTEQDSRDSTYSLSSTLTLSRADYER HRVYACEVTHQGLS S P

- 131 040365

SEQ ID NO: 126 DNA, light chain GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACCGAGTGACAATCA CCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAGCCTGATGCAGTGGTATCAGCAAAA GCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCA AGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGG ACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAA AGTCGAAATTAAACGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAA CTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGG TCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGA CTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAG CACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCA ACAGAGGAGAATGT ABTIM3-hum20 SEQ ID NO: 3 (Kabat) HCDR1 SYNMH SEQ ID NO: 30 (Kabat) HCDR2 DIYPGQGDTSYNQKFKG SEQ ID NO: 5 (Kabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 31 (Chothia) HCDR2 YPGQGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 80 vh EVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIYPGQGDTSYNQKF KGQVTISADKSIS TVYLQWS S LKAS DTAMYYCARVGGAFPMDYWGQGT TVTVSS SEQ ID NO: 81 DNA, VH GAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTTTGAAAATTTCCT GCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCACTGGGTGAGACAGATGCCAGGCAA GGGCCTGGAGTGGATGGGAGACATATACCCAGGCCAGGGAGACACAAGCTATAATCAGAAATTC AAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCAC TTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTA TTGGGGCCAAGGGACCACAGTAACCGTCAGCTCA SEQ ID NO: 82 heavy chain EVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIYPGQGDTSYNQKF KGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPE VTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCK VSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

- 132 040365

DNA, GAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTTTGAAAATTTCCT GCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCACTGGGTGAGACAGATGCCAGGCAA GGGCCTGGAGTGGATGGGAGACATATACCCAGGCCAGGGAGACACAAGCTATAATCAGAAATTC AAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCAC TTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTA TTGGGGCCAAGGGACCACAGTAACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCA CTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACT ACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTT CCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGC TCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATA SEQ ID NO: 83 heavy chain AAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGG ACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAA GTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGG ACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAG GGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAG GTGTCCAACAAAGGACTGCCAAGCTCTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTA GAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTT GACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAG CCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATT CCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCA CGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 (Rabat) LCDR1 RASESVEYYGTSLMQ SEQ ID NO: 7 (Rabat) LCDR2 AASNVES SEQ ID NO: 8 (Rabat) SEQ ID NO: 12 (Chothia) SEQ ID NO: 13 (Chothia) SEQ ID NO: 14 (chothia) SEQ ID NO: 64 LCDR3 LCDR1 LCDR2 LCDR3 VL QQSRRDPST SESVEYYGTSL AAS SRRDPS AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPRLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGGGTKVEIR GCAATACAGTTGACACAGAAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACCGAGTGACAATCA CCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAGCCTGATGCAGTGGTATCAGCAAAA GCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCA SEQ ID NO: 125 SEQ ID NO: 66 DNA, VL light chain AGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTTCAGCCAGAGG ACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGTGGGACAAA AGTCGAAATTAAA AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPRLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLEC FRGNSVTHQGL SRVYACEVTHQGL

- 133 040365

SEQ ID NO: 126 DNA, light chain GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACCGAGTGACAATCA CCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAGCCTGATGCAGTGGTATCAGCAAAA GCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCA AGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGG ACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAA AGTCGAAATTAAACGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAA CTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGG TCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGA CTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAG CACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCA ACAGAGGAGAATGT ABTIM3-hum21 SEQ ID NO: 3 (Rabat) HCDR1 SYNMH SEQ ID NO: 30 (Rabat) HCDR2 DIYPGQGDTSYNQRFRG SEQ ID NO: 5 (Rabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 31 (Chothia) HCDR2 YPGQGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 84 vh QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIYPGQGDTSYNQKF KGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSS SEQ ID NO: 85 DNA, VH CAGGTGCAATTGGTGCAGAGCGGAGCAGAGGTCAAAAAGCCCGGAGCAAGCGTGAAGGTCTCAT GCAAAGCAAGCGGATACACATTTACATCATACAACATGCACTGGGTCAGGCAGGCTCCAGGACA GGGACTGGAGTGGATCGGGGACATCTACCCTGGACAGGGCGATACTAGCTATAATCAGAAGTTC AAAGGCCGGGCCACCATGACAGCTGACAAGTCTACTAGTACCGTGTATATGGAACTGAGCTCCC TGCGGTCTGAAGATACCGCAGTGTACTATTGCGCCAGAGTCGGGGGGGCATTTCCTATGGATTA TTGGGGGCAGGGGACTCTGGTCACTGTCAGCTCA SEQ ID NO: 86 heavy chain QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIYPGQGDTSYNQKF KGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTRGPSVFP LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPRDTLMISR TPEVTCVWAVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEY RCKVSNKALAAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVRGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

- 134 040365

SEQ ID NO: 87 DNA, heavy chain CAGGTGCAATTGGTGCAGAGCGGAGCAGAGGTCAAAAAGCCCGGAGCAAGCGTGAAGGTCTCAT GCAAAGCAAGCGGATACACATTTACATCATACAACATGCACTGGGTCAGGCAGGCTCCAGGACA GGGACTGGAGTGGATCGGGGACATCTACCCTGGACAGGGCGATACTAGCTATAATCAGAAGTTC AAAGGCCGGGCCACCATGACAGCTGACAAGTCTACTAGTACCGTGTATATGGAACTGAGCTCCC TGCGGTCTGAAGATACCGCAGTGTACTATTGCGCCAGAGTCGGGGGGGCATTTCCTATGGATTA TTGGGGGCAGGGGACTCTGGTCACTGTCAGCTCAGCTAGCACCAAGGGCCCCAGCGTGTTCCCC CTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACT ACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGCGTGCACACCTT CCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGC AGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACA AGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCAGAGCT GCTGGGCGGACCCTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGG ACCCCCGAGGTGACCTGCGTGGTGGTGGCCGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACT GGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAG CACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAATAC AAGTGCAAGGTCTCCAACAAGGCCC TGGCAGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGG GCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCTCCCGGGAGGAGATGACCAAGAACCA GGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGC AACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCT TCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAG CGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTCCCCCGGCAAG SEQ ID NO: 6 (Rabat) LCDR1 RASESVEYYGTSLMQ SEQ ID NO: 8 (Rabat) LCDR3 QQSRRDPST SEQ ID NO: 12 (Chothia) LCDR1 SESVEYYGTSL SEQ ID NO: 13 (Chothia) LCDR2 AAS SEQ ID NO: 14 (Chothia) LCDR3 SRRDPS SEQ ID NO: 88 VL DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPRLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGGGTRVEIR SEQ ID NO: 89 DNA, VL GACATCGTCCTGACACAAGTCTCCTGACAGCCTGGCAGTGAGCCTGGGCGAAAGGGCAACCATTA ATTGTAGAGCCTTCCGAGTCCGTCGAGTACTATGGCACTAGTCTGATGCAGTGGTACCAGCAGAA GCCAGGGCAGCCCCCTAAACTGCTGATCTATGCAGCTAGCAACGTGGAGTCCGGAGTCCCAGAC CGGTTCTCTGGAAGTGGGTCAGGAACCGATTTTACCCTGACAATTAGCTCCCTGCAGGCAGAAG ACGTGGCCGTCTACTATTGTCAGCAGAGCCGCAAGGACCCAAGCACATTCGGAGGGGGGACCAA AGTGGAAAATCAAG SEQ ID NO: 90 Light chain DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPRLLIYAASNVESGVPD RFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTTFGGGTKVEIRRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKECDSTYSLSSTLTLSRADYEK HRVYACEVRS

- 135 040365

SEQ ID NO: 91 DNA, light chain GACATCGTCCTGACACAGTCTCCTGACAGCCTGGCAGTGAGCCTGGGCGAAAGGGCAACCATTA ATTGTAGAGCTTCCGAGTCCGTCGAGTACTATGGCACTAGTCTGATGCAGTGGTACCAGCAGAA GCCAGGGCAGCCCCCTAAACTGCTGATCTATGCAGCTAGCAACGTGGAGTCCGGAGTCCCAGAC CGGTTCTCTGGAAGTGGGTCAGGAACCGATTTTACCCTGACAATTAGCTCCCTGCAGGCAGAAG ACGTGGCCGTCTACTATTGTCAGCAGAGCCGCAAGGACCCAAGCACATTCGGAGGGGGGACCAA AGTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAG CTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGG TGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGA CAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAG CATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCA ACAGGGGCGAGTGC ABTIM3-hum22 SEQ ID NO: 3 (Rabat) HCDR1 SYNMH SEQ ID NO: 30 (Rabat) HCDR2 DIYPGQGDTSYNQRFRG SEQ ID NO: 5 (Rabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 31 (Chothia) HCDR2 YPGQGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 92 vh QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGQGDTSYNQKF RGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSS SEQ ID NO: 93 DNA, VH CAGGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAACCAGGCGCCAGCGTGAAGGTGTCCT GCAAGGCCAGCGGCTACACCTTTACCAGCTACAACATGCACTGGGTGCGCCAGGCCCCTGGACA GGGACTGGAATGGATGGGCGACATCTACCCCGGCCAGGGCGACACCTCCTACAACCAGAAATTC AAGGGCAGAGTGACCATGACCCGGGACACCAGCACCTCCACCGTGTACATGGAACTGAGCAGCC TGCGGAGCGAGGACACCGCCGTGTACTACTGTGCTAGAGTGGGCGGAGCCTTCCCCATGGACTA TTGGGGCCAGGGCACCACCGTGACCGTGAGCTCA SEQ ID NO: 94 heavy chain QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGQGDTSYNQKF RGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSSASTRGPSVFP LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPRDTLMISR TPEVTCVWAVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEY RCKVSNKALAAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVRGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

- 136 040365

SEQ ID NO: 95 DNA, heavy chain CAGGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAACCAGGCGCCAGCGTGAAGGTGTCCT GCAAGGCCAGCGGCTACACCTTTACCAGCTACAACATGCACTGGGTGCGCCAGGCCCCTGGACA GGGACTGGAATGGATGGGCGACATCTACCCCGGCCAGGGCGACACCTCCTACAACCAGAAATTC AAGGGCAGAGTGACCATGACCCGGGACACCAGCACCTCCACCGTGTACATGGAACTGAGCAGCC TGCGGAGCGAGGACACCGCCGTGTACTACTGTGCTAGAGTGGGCGGAGCCTTCCCCATGGACTA TTGGGGCCAGGGCACCACCGTGACCGTGAGCTCAGCTAGCACCAAGGGCCCCAGCGTGTTCCCC CTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACT ACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGCGTGCACACCTT CCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGC AGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACA AGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCAGAGCT GCTGGGCGGACCCTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGG ACCCCCGAGGTGACCTGCGTGGTGGTGGCCGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACT GGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAG CACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAATAC AAGTGCAAGGTCTCCAACAAGGCCC TGGCAGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGG GCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCTCCCGGGAGGAGATGACCAAGAACCA GGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGC AACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCT TCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAG CGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTCCCCCGGCAAG SEQ ID NO: 6 (Kabat) SEQ ID NO: 7 (Kabat) SEQ ID NO: 8 (Kabat) SEQ ID NO: 12 (Chothia) SEQ ID NO: 13 (Chothia) SEQ ID NO: 14 (Chothia) SEQ ID NO: 96 SEQ ID NO: 97 SEQ ID NO: 98 LCDR1 LCDR3 LCDR1 LCDR2 LCDR3 VL DNA, VL light chain RASESVEYYGTSLMQ ^AASNVES QQSRKDPST SESVEYYGTSL AAS SRKDPS AIRLTQSPSSFSASTGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQSEDFATYYCQQSRKDPSTFGGGTKVEIK GCCATCAGACTGACCCAGAGCCCCAGCTCCTTTAGCGCCAGCACCGGCGACAGAGTGACCATCA CCTGTAGAGCCAGCGAGAGCGTGGAATATTACGGCACCAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGCAAGGCCCCCAAGCTGCTGATCTACGCCGCCAGCAATGTGGAAAGCGGCGTGCCCAGC AGATTCAGCGGCTCTGGCAGCGGCACCGACTTCACCCTGACAATCAGCAGCCTGCAGAGCGAGG ACTTCGCCACCTACTACTGCCAGCAGAGCCGGAAGGACCCCAGCACATTTGGCGGAGGCACCAA GGTGGAAATCAAG AIRLTQSPSSFSASTGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQSEDFATYYCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSKDSRGSLSSSTLTLSKADYEKS HKGLVYACEV

- 137 040365

SEQ ID NO: 99 DNA, light chain GCCATCAGACTGACCCAGAGCCCCAGCTCCTTTAGCGCCAGCACCGGCGACAGAGTGACCATCA CCTGTAGAGCCAGCGAGAGCGTGGAATATTACGGCACCAGCCTGATGCAGTGGTATCAGCAGAA GCCCGGCAAGGCCCCCAAGCTGCTGATCTACGCCGCCAGCAATGTGGAAAGCGGCGTGCCCAGC AGATTCAGCGGCTCTGGCAGCGGCACCGACTTCACCCTGACAATCAGCAGCCTGCAGAGCGAGG ACTTCGCCACCTACTACTGCCAGCAGAGCCGGAAGGACCCCAGCACATTTGGCGGAGGCACCAA GGTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAG CTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGG TGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGA CAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAG CATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCA ACAGGGGCGAGTGC ABTIM3-hum23 SEQ ID NO: 3 (Rabat) HCDR1 SYNMH SEQ ID NO: 4 (Rabat) HCDR2 DIYPGNGDTSYNQRFRG SEQ ID NO: 5 (Rabat) HDDR3 VGGAFPMDY SEQ ID NO: 9 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 10 (Chothia) HCDR2 YPGNGD SEQ ID NO: 5 (Chothia) HDDR3 VGGAFPMDY SEQ ID NO: 100 vh QVQLVQSGAEVKRPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIYPGNGDTSYNQRF RGQVTISADRSIS TVYLQWS S LRAS DTAMYYCARVGGAFPMDYWGQGT TVTVS S SEQ ID NO: 101 DNA, VH CAGGTGCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTTTGAAAATTTCCT GCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCACTGGGTGAGACAGATGCCAGGCAA GGGCCTGGAGTGGATGGGAGACATATACCCAGGCAATGGAGACACAAGCTATAATCAGAAATTC AAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCAC TTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTA TTGGGGCCAAGGGACCACAGTAACCGTCAGCTCA SEQ ID NO: 102 heavy chain QVQLVQSGAEVKRPGESLRISCRGSGYTFTSYNMHWVRQMPGRGLEWMGDIYPGNGDTSYNQRF RGQVTISADRSISTVYLQWSSLRASDTAMYYCARVGGAFPMDYWGQGTTVTVSSASTRGPSVFP LAPSSRSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVWAVSHEDPEVRFNWYVDGVEVHNARTRPREEQYNSTYRWSVLTVLHQDWLNGREY RCRVSNRALAAPIERTISRARGQPREPQVYTLPPSREEMTRNQVSLTCLVRGFYPSDIAVEWES NGQPENNYRTTPPVLDSDGSFFLYSRLTVDRSRWQQGNVFSCSVMHEALHNHYTQRSLSLSPGR

- 138 040365

SEQ ID NO: 103 DNA, heavy chain CAGGTGCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTTTGAAAATTTCCT GCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCACTGGGTGAGACAGATGCCAGGCAA GGGCCTGGAGTGGATGGGAGACATATACCCAGGCAATGGAGACACAAGCTATAATCAGAAATTC AAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCAC TTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTA TTGGGGCCAAGGGACCACAGTAACCGTCAGCTCAGCTAGCACCAAGGGCCCCAGCGTGTTCCCC CTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACT ACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGCGTGCACACCTT CCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGC AGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACA AGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCAGAGCT GCTGGGCGGACCCTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGG ACCCCCGAGGTGACCTGCGTGGTGGTGGCCGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACT GGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAG CACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAATAC AAGTGCAAGGTCTCCAACAAGGCCC TGGCAGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGG GCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCTCCCGGGAGGAGATGACCAAGAACCA GGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGC AACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCT TCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAG CGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTCCCCCGGCAAG SEQ ID NO: 6 (Rabat) SEQ ID NO: 8 (Rabat) SEQ ID NO: 12 (Chothia) SEQ ID NO: 13 (Chothia) SEQ ID NO: 14 (Chothia) SEQ ID NO: 104 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 107 LCDR1 LCDR3 LCDR1 LCDR2 LCDR3 VL DNA, VL light chain DNA, light chain RASESVEYYGTSLMQ QQSRRDPST SESVEYYGTSL AAS SRRDPS AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQRPGRAPRLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQPEDFATYFCQQSRRDPSTFGGGTRVEIR GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACCGAGTGACAATCA CCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAGCCTGATGCAGTGGTATCAGCAAAA GCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCA AGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGG ACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAA AGTCGAAATTAAA AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQRPGRAPRLLIYAASNVESGVPS RFSGSGSGTDFTLTISSLQPEDFATYFCQQSRRDPSTFGGGTRVEIRRTVAAPSVFIFPPSDEQ LRSGTASWCLLNNFYPREARVQWRVDNALQSGNSQESVTEQDSRDSTYSLSSTLTLSRADYER HRVYACEVTHQGLS S P gcaatacagttgacacagagtccttcaagtttgtccgcttccgttggcgaccgagtgacaatca cctgtagagcatccgagtcagtggagtattatggcactagcctgatgcagtggtatcagcaaaa gccagggaaagccccaaagctgctgatatatgccgcgagtaacgtcgagtcaggggtgccatca agattctccggttccgggtccggaaccgacttcacactgaccatctcttcccttcagccagagg acttcgctacgtacttttgccagcagtcacggaaagatccctctactttcggaggtgggacaaa agtcgaaattaaacgtacggtggccgctcccagcgtgttcatcttcccccccagcgacgagcag ctgaagagcggcaccgccagcgtggtgtgcctgctgaacaacttctacccccgggaggccaagg tgcagtggaaggtggacaacgccctgcagagcggcaacagccaggagagcgtcaccgagcagga cagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaag cataaggtgtacgcctgcgaggtgacccaccagggcctgtccagccccgtgaccaagagcttca acaggggcgagtgc

- 139 040365

Table 5. Amino acid sequences of the constant region of human IgG heavy chains and human kappa light chains.

ns Amino acid sequence of mutant IgG4 constant region (S228P) (EU numbering) ASTKGPSVFP LAPCSRSTSE STAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS GLYSLSSWT VPSSSLGTKT YTCNVDHKPS NTKVDKRVES KYGPPCPPCP APEFLGGPSV FLFPPKPKDT LMISRTPEVT CVWDVSQED PEVQFNWYVD GVEVHNAKTK PREEQFNSTY RWSVLTVLH QDWLNGKEYK CKVSNKGLPS SIEKTISKAK GQPREPQVYT LPPSQEEMTK NQVSLTCLVK GFYPSDIAVE WESNGQPENN YKTTPPVLDS DGSFFLYSRL TVDKSRWQEG NVFSCSVMHE ALHNHYTQKS LSLSLGK (SEQ ID NO: 108) LC Amino acid sequence of human kappa constant region RTVAAPSVFI FPPSDEQLKS GTASWCLLN NFYPREAKVQ WKVDNALQSG NSQESVTEQD SKDSTYSLSS TLTLSKADYE KHKVYACEVT HQGLSSPVTK SFNRGEC (SEQ ID NO: 109) NS Amino acid sequence of mutant IgG4 constant region (S228P) lacking C-terminal lysine (K) (EU numbering) ASTKGPSVFP LAPCSRSTSE STAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS GLYSLSSWT VPSSSLGTKT YTCNVDHKPS NTKVDKRVES KYGPPCPPCP APEFLGGPSV FLFPPKPKDT LMISRTPEVT CVWDVSQED PEVQFNWYVD GVEVHNAKTK PREEQFNSTY RWSVLTVLH QDWLNGKEYK CKVSNKGLPS SIEKTISKAK GQPREPQVYT LPPSQEEMTK NQVSLTCLVK GFYPSDIAVE WESNGQPENN YKTTPPVLDS DGSFFLYSRL TVDKSRWQEG NVFSCSVMHE ALHNHYTQKS LSLSLG (SEQ ID NO: 110) NS wild-type IgGl ASTKGPSVFP LAPSSKSTSG GTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS GLYSLSSWT VPSSSLGTQT YICNVNHKPS NTKVDKRVEP KSCDKTHTCP PCPAPELLGG PSVFLFPPKP KDTLMISRTP EVTCVWDVS HEDPEVKFNW YVDGVEVHNA KTKPREEQYN STYRWSVLT VLHQDWLNGK EYKCKVSNKA LPAPIEKTIS KAKGQPREPQ VYTLPPSREE MTKNQVSLTC LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SKLTVDKSRW QQGNVFSCSV MHEALHNHYT QKSLSLSPGK (SEQ ID NO: 111)

- 140 040365

ns Amino acid sequence of mutant IgGl constant region (N297A) (EU numbering) ASTKGPSVFP LAPSSKSTSG GTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS GLYSLSSWT VPSSSLGTQT YICNVNHKPS NTKVDKRVEP KSCDKTHTCP PCPAPELLGG PSVFLFPPKP KDTLMISRTP EVTCVWDVS HEDPEVKFNW YVDGVEVHNA KTKPREEQYA STYRWSVLT VLHQDWLNGK EYKCKVSNKA LPAPIEKTIS KAKGQPREPQ VYTLPPSREE MTKNQVSLTC LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SKLTVDKSRW QQGNVFSCSV MHEALHNHYT QKSLSLSPGK (SEQ ID NO: 112) NS Amino acid sequence of the mutant IgGl constant region (D265A, P329A) (EU numbering) ASTKGPSVFP LAPSSKSTSG GTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS GLYSLSSWT VPSSSLGTQT YICNVNHKPS NTKVDKRVEP KSCDKTHTCP PCPAPELLGG PSVFLFPPKP KDTLMISRTP EVTCVWAVS HEDPEVKFNW YVDGVEVHNA KTKPREEQYN STYRWSVLT VLHQDWLNGK EYKCKVSNKA LAAPIEKTIS KAKGQPREPQ VYTLPPSREE MTKNQVSLTC LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SKLTVDKSRW QQGNVFSCSV MHEALHNHYT QKSLSLSPGK (SEQ ID NO: 113) NS Amino acid sequence of the mutant IgGl constant region (L234A, L235A) (EU numbering) ASTKGPSVFP LAPSSKSTSG GTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS GLYSLSSWT VPSSSLGTQT YICNVNHKPS NTKVDKRVEP KSCDKTHTCP PCPAPEAAGG PSVFLFPPKP KDTLMISRTP EVTCVWDVS HEDPEVKFNW YVDGVEVHNA KTKPREEQYN STYRWSVLT VLHQDWLNGK EYKCKVSNKA LPAPIEKTIS KAKGQPREPQ VYTLPPSREE MTKNQVSLTC LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SKLTVDKSRW QQGNVFSCSV MHEALHNHYT QKSLSLSPGK (SEQ ID NO: 114)

Table 6. Selected drugs that can be administered in combination with anti-TIM-3 antibody molecules, eg, alone or in combination with other immunomodulators described herein. Each publication listed in this table is incorporated into the present description by reference in its entirety, including all structural formulas in it. ___________________________________________________________

Compound No. generic name, trade name Connection structure Patents/publications patent applications Al Sotrastaurin K th O CH ₃ EP 1682103 US 2007/142401 WO2005/039549 A2 Nilotinib HC1 monohydrate, TASIGNA® LH _d at ctYMV N -D HC1 H ₂ O WO2004/005281 US 7169791 A3 H ₃ C but for / .n 1 -o WO 2010/060937 WO2004/072051 EP 1611112 US 8450310

- 141 040365

A4 Dactolisib N. . co » ί ί! . Mr-sn, • 8 Ί - N WO 2006/122806 A5 ₀ -GH _J ϋ—(, ^Cl H,C \—( сГ NH Ν-ΓΗΝ V ./7 ν— ; ΝΗ 0 ο Ν—/ <CH Cog US 8552002 Ab Buparlisib 1°: Ν' GU ^X, ''O Η _: Ν Ν WO2007/084786

- 142 040365

_CH30CL A7 'NII WO 2009/141386, US 2010/0105667 ΗΝΥ A' ) H,CN ₄ CH ₃ A8 ^MC <r“ »□ __ 0 i . “A o^nh ₂ bV WO 2010/029082 K ., n /. ' WO 2010/149755 US A9 CYP17 inhibitor 8263635 B2, EP 2445903 Bl AYu SN ₃ °y^ and O. f l . ^r “S r, ; 0 W02011/076786 1 1 b 1 N _s ί H C OH sn [CI Deferasirox, O But / All EXJADE® WO 1997/049395 t - 1 ^! · but he

- 143 040365

A12 Lectrozole, FEMARA® N^. II i 1 US 4978672 .ABOUT. 'Ί A13 LJ F “ T ϊ 4 r ^0H N ']' N ' ^ _tll< J', H,NN n WO 2013/124826, US 2013/0225574 Cl Λ .CH, γ^ 'N ³ Cl Ύ^° A14 ^vLT_N WO 2013/111105 HjC j Q r T ^ХНз ] About H _? C CH ₃ HN 0 A15 0 У ^N γ ⁵ WO2005/073224 _rNH

- 144 040365

A16 Imatinib mesylate, GLEEVEC® CH, N N-----' / ί, l 'A // 'r---O ΗΝ o ΗΝ ^Ζ bn ₃ -O η - -Ν Mesylate WO 1999/003854 A17 N-S Nil 0=6F — Ν' N N-Ch N / NJ Dihydrochloric acid EP2099447, US 7767675, US 8420645 A18 Ruxolitinib phosphate, JAKAFI® L / N N - N ^N ' O ' m- N N _H H ₃ PO ₄ W02007/070514EP US 2474545 7598257, WO 2014/018632 A19 Panobinostat ABOUT lL ,0H 'A 0 Y J 'h ™4„. WO 2014/072493, WO2002/022577, EP 1870399 A20 Osilodrostat / , 2^^ > WO2007/024945 A21 Y WO 2008/016893, EP 2051990, US 8546336

- 145 040365

A22 Sonidegiba phosphate ^F . 1 ^fA L •ΰ < сн ₅ \=0 nm'< Vcm Η Nzb*n WO 2007/131201, EP 2021328, US 8178563 A2 3 Seritinib, ZYKADIA™ “r, L ¹ ' Ν Ν Ή' A ο=έ=α ^{Η n} 1 Г WO 2008/073687, US 8039479 A24 “ with ~ H1H Ν N^UH. L 'U- _r ,Ν f 1 'N'' Η US 8415355, US 8685980 A25 CH, Η-,οψοίΙ φ ^iC ?: WO 2010/007120 A26 Human monoclonal antibody to PRLR US 7867493

- 146 040365

A27 HN *CH. ' Ί g ^F ! ; Ί _th ή L ί o n WO 2010/026124, EP 2344474, US 2010/0056576, WO2008/106692 A2 8 ^=\ ⁿ ' ^with A ) SI·, mC. 0^ ΝΗ jl Ν Ν WO2010/101849 A29 Encorafenib o th , ^H ' Π ΝΗ 0 sn, | Ν'*''Ν Id, οι FIN—\ H , -fl ™ °7 A o ^S CH V Cl W02011/025927 AZO N'< ^N - ^CH ' ί! I HN 'Ν' Ν Π "J /.¾ ⁰ 'ν-A' H _a C WO2011/101409 A31 monoclonal antibody human to HER3 WO 2012/022814, EP 2606070, US 8735551

- 147 040365

A32 Antibody-conjugate medicinal product (ADC) WO 2014/160160 Ab: 12425 (see Table 1, paragraph [00191]) Linker: SMCC (see Paragraph [00117] Cargo: DM1 (see Paragraph [00111] See also claim 29 of the claims AZZ Monoclonal antibody or Fab to M-CSF WO2004/045532 AZ 4 Binimetinib H ISK N„.O ° H ί WO2003/077914 H ₃ C- _N ^Z AZ 5 Midostaurin _x g ' ί z 1 1 w ^- oil o I WO 2003/037347, EP 1441737, US 2012/252785 NCH, OH 0—/ / / VQ' /---/ ^X CH ₃ AZ 6 Everolimus, AFINITOR® \ WCH ₃ / ⁿ ₃ Χ^-γ ⁰ «Vo ⁷ HjC but ^=θ °^\ Ο χ ^Η n _e s 7==/ n ₃ s WO 2014/085318

- 148 040365

AZ 7 'p Xj ~ ^F h WO 2007/030377, US 7482367 N— Χν ΗΝ^ X pasireotida _Η J \ , WO2002/010192, AZ 8 diaspartate, SIGNIFOR® 4n ^! Χ , ^Γ W - 1 1! . US 7473761 XX ΗΝ J^NH _? AZ 9 dovitinib <Χ\ 'ΝΗ N 1 1 Ί WO 2009/115562, US 8563556 Λ N CH, A4 0 I' J HC— . “ D Ν— ά ^Ν ,ί H-c C... WO 2013/184757

- 149 040365

A41 HN- , _F p If 7 ,, , a N ,3 ^ΗΛ ( J, Γν.„, [1 J '1 ,- N WO 2006/122806 A42 CH _S A [ J ^ΗΛ - CH 1 4 i ¹ η ' ό -441, N NH jJ UN θ L fl H cs - T o Git WO 2008/073687, US 8372858 A43 H N._CH _a ,,=/' V N VNH ^N HN 0 h 0 WO 2010/002655, US 8519129 A44 0-^ <> H ₃ c '^-F NH KN—N ci H _a C -N' H WO 2010/002655, US 8519129

- 150 040365

A4 5 N.S-l N - _h / 7 H ₃ C - VF ΝΗ Ν \u003d ΐ 4 Ν Gl' ΝΗ Μ' ΗΝ. z ^! λ. W02010/002655 A4 6 Valspodar, AMDRAY™ ^Ν !<^ S ^Ν b Η < ^Ν ,.. ^! EP 296122 A4 7 Watalaniba succinate v Succinate W098/35958 A4 8 IDH inhibitor WO2014/141104 A4 9 BCR-ABL inhibitor WO2013/171639 WO2013/171640 WO2013/171641 WO2013/171642 A5 0 cRAF inhibitor WO2014/151616 A51 Competitive inhibitor of ERK1/2 ATP PCT/US2014/062913

Table 7. See examples.

Table 8. See examples.

Table 9. See examples.

Table 10. See examples.

Table 11. See examples.

Table 12. See examples.

Table 13. See examples.

Table 14. See examples.

Table 15. See examples.

Table 16. See examples.

Examples

Example 1 Characterization of ABTIM3 and Other Anti-TIM-3 Antibodies.

Anti-TIM-3 antibody panels were analyzed for binding to TIM-3 expressing cells. The dissociation constants (KD) of two such antibodies, ABTIM3 and anti-TIM-3 antibody #2, as measured using surface plasmon resonance, are summarized in FIG. 2A. HA fig. 2B and 2C, the binding of various anti-TIM-3 antibodies, including ABTIM3, to cells transfected with human TIM-3 was measured using flow cytometry. Further, three antibodies: ABTIM3, anti-TIM-3 antibody #2 and anti-TIM-3 antibody #3, and a control antibody were analyzed for the ability to bind cynomolgus TIM-3 in cells transfected with cynomolgus TIM-3. In FIG. 2D shows that of the three antibodies tested, ABTIM3 has the highest affinity for cynomolgus TIM-3. In FIG. 2E shows a study of seven anti-human TIM-3 antibodies for binding to cynomolgus monkey TIM-3 and shows that ABTIM3 binds with the highest affinity. Overall, experiments indicate that ABTIM3 has a high (subnanomolar) affinity for both human and Javanese TIM-3.

- 151 040365 macaque.

The ability of three anti-TIM-3 antibodies, including ABTIM3, to bind to human TIM-4 expressed in CHO cells and mouse TIM-3 expressed in cells was also measured by flow cytometry. Human TIM-3 shares approximately 23% sequence identity with human TIM-4. Mouse TIM-3 shares approximately 66% sequence identity with human TIM-3 and 64% sequence identity with cynomolgus TIM-3. The results of these assays demonstrate that ABTIM3 does not bind to human TIM-4. ABTIM3 also does not cross-react with mouse TIM-3. In conjunction with the results of the binding assay described above, the ABTIM3 antibody is specific for human and cynomolgus TIM-3.

In a cross-blocking experiment, ABTIM3 was shown to cross-block anti-TIM-3 antibody #2, indicating that these antibodies bind to epitopes that are close to each other and possibly overlap, although the two epitopes are not necessarily identical. .

The ability of anti-TIM-3 antibodies, such as ABTIM3, to bind to activated PBMCs expressing TIM-3 was also evaluated. Human whole blood PBMCs from a donor were stimulated for 10 days with plate-bound CD3/CD28 (1 μg/ml each) in the presence of 10 ng/ml recombinant human IL-12. Cells were treated with ficoll to remove dead cells and reactivated for three days with the same stimulus. Antibodies that bind to TIM-3, such as ABTIM3 and anti-TIM-3 antibody #2, were compared and anti-PD-1, anti-PD-L1, and anti-LAG-3 and mouse IgG1 antibodies were used as control antibodies. Cells were incubated with antibodies at various concentrations from 0.001 to 100 μg/ml and antibody binding to cells was analyzed by flow cytometry. Cells were restricted to CD4 positive or CD8 populations and the mean fluorescence intensity (MFI) for each antibody and concentration was plotted. Then the values of the dissociation constant (K _D ) were calculated. The results of the analyzes are presented in table. 7 below.

Table 7. K _D values for anti-TIM-3 antibody binding to activated PBMCs.

Antibody CD4-restricted RVMS, K _D CD8-restricted PBMCs, K _D ABTIM3 0.2 9 nM 0.30 nM Antibody against TIM-3 #2 2.84 nM 3.14 nM Anti-PD-antibody L1, control 0.2 0 nM 0.30 nM Antibody against LAG-3, control 2.33 nM Anti-PD-antibody 1, control 22.8 nM 8 5.9 nM

These results demonstrate that ABTIM3 was able to bind to TIM-3 expressed on activated PBMCs.

Example 2 Analysis of Anti-TIM-3 Antibody Domains Binding to TIM-3.

TIM-3 has an extracellular IgV domain and a mucin domain. The TIM-3 regions bound by each of the five antibodies were determined using a recombinant construct that replaced the TIM-3 IgV domain with the PD-1 IgV domain, and this construct is shown in FIG. 3A. In FIG. 3B shows that an anti-TIM-3 monoclonal antibody (anti-TIM-3 antibody #3) and two control anti-PD-L1 monoclonal antibodies (anti-PD-L1 antibodies #1 and #2) bind to the chimeric protein shown in FIG. . 3A, while anti-TIM-3 #2 and ABTIM3 do not substantially bind. This result indicates that the anti-TIM-3 monoclonal antibodies, anti-TIM-3 antibody #2 and ABTIM3, bind to the IgV domain of TIM-3, while anti-TIM-3 antibody #3 binds to the mucin domain of TIM-3. 3. The values of dissociation constants (K _D ), calculated for each test antibody in relation to the recombinant construct, are presented in table. 8.

- 152 040365

Table 8. KD values for binding to the IgV PD-L1 construct/TIM-3 mucin domain.

Antibody Antigen K _D Antibody against PD-L1 #1 IgV domain PD-L1 0.52 nM Antibody against PD-L1 #2 IgV domain PD-L1 0.38 nM Antibody against TIM-3 #3 TIM-3 mucin domain 2.71 nM Antibody against TIM-3 #2 TIM-3 No binding to chimeric protein ABTIM3 TIM-3 No binding to chimeric protein

Example 3 TIM-3 binding to PtdSer is blocked by anti-TIM-3 antibodies.

TIM-3 binds to PtdSer (phosphatidylserine) lipid, which is normally present on the surface of apoptotic cells but not normal cells. WR19L(Fas) cells treated with anti-CD95 antibody were cultured under conditions that stimulate the accumulation of PtdSer on the cell surface (movement of PtdSer from the inner to the outer membrane upon induction of apoptosis). TIM-3-Ig (huTIM-3 extracellular domain fused to Ig Fc region) was added to the cells, and TIM-3-Ig binding to the cells was analyzed in the presence of various antibodies. As shown in FIG. 4, several anti-TIM-3 mAbs, including ABTIM3, anti-TIM-3 #5, and anti-TIM-3 #2, inhibit TIM-3 binding to PtdSer.

Example 4 Secretion of IFN-gamma by CD4+ cells is enhanced by antibodies against TIM-3.

The ability of four antibodies to enhance IFN-gamma secretion and proliferation of IL-12 stimulated CD4+ cells was analyzed. This analysis utilized the observation that a high dose of IL-12 induces TIM-3 expression and produces a depleted T cell phenotype (see Yang et al., J. Clin. Invest. 122:4 p1271 2012). In FIG. 5A shows four panels, each indicating the results of an experiment where cells were exposed to an antibody selected from ABTIM3, anti-TIM-3 #2, mIgG1, and anti-PD-L1 (control anti-PD-L1). After restimulation with PMA/ionomycin and fixation and cell permeability, the resulting IFNgamma levels were measured by flow cytometry (y-axis) and proliferation was measured by CFSE fluorescence (x-axis). In FIG. 5B quantifies IFN-gamma expression in cells exposed to these four antibodies. From left to right, the bars in Fig. 5B correspond to ABTIM3 antibody, anti-TIM-3 antibody #2, control anti-PD-L1 antibody and mIgG1.

Example 5 Blockade of TIM-3 enhances functional activity in vitro.

5.1. Blockade of TIM-3 enhances the in vitro cytotoxic activity of purified NK cells

TIM-3 is endogenously expressed at a high level on NK cells (natural killer cells); its expression is furthermore induced on activated NK cells. TIM-3 can act to restore NK cell function like other inhibitory receptors. See Ndhlovu et al., Blood 119:3734, 2012, and Silva et al., Cancer Immunol Res 2:410, 2014. The ability of ABTIM3 and other anti-TIM-3 antibodies to enhance the cytotoxic activity of NK cells was thus evaluated.

In this assay, NK cells were purified from whole blood by negative selection on beads and then incubated with antibody (10 μg/ml) at 37°C. After 1 h, K562 target cells were added. After incubation for 4 hours at 37°C, the percentage kill of K562 cells was measured. The ABTIM3 antibody resulted in increased levels of K523 cell killing relative to the anti-TIM-3 antibody #2 or isotype control.

5.2. TIM-3 Blockade Increases Proliferation in Autologous T-DC Cocultures

TIM-3 can be expressed on dendritic cells (DC) and T cells. Naive T cells and dendritic cells were isolated from donor samples. Naive T cells and conventional DCs were co-cultured for four days in the presence of an anti-CD3/CD28 antibody. ABTIM3 was added to the coculture at various doses: 0.01 µg/ml, 0.1 µg/ml, 1 µg/ml, 5 µg/ml and 25 µg/ml. Cell proliferation was detected by a CFSE proliferation assay, which is based on a dilution of the CFSE dye to detect proliferating cells.

As shown in FIG. 22, the presence of ABTIM3 at each dose tested resulted in an increase in the number of proliferating cells, as indicated by cells with diluted CFSE, compared with no antibody and mouse isotype control (IgG1).

Example 6 Characterization of a humanized anti-TIM-3 antibody.

6.1. Obtaining humanized antibodies against TIM-3

The mouse anti-TIM-3 antibody ABTIM3 was humanized by transplantation of CDRs such as those shown in Table 1. 3 into a hinge-stabilized human IgG4 constant region containing the S228P mutation. Additional modifications were made to CDR2 of the heavy chain pu

- 153 040365 by mutating the putative deamidation site with N at position 6 of HCDR (Kabat) or position 4 of HCDR2 (Chothia) at S or Q to eliminate the deamidation site. Other modifications included the use of alternative wireframe regions. Unique heavy chains and light chains were combined in various combinations to form a small library of unique humanized mAbs.

6.2. Binding assays

The binding capacity of the resulting humanized mAbs was examined using parental mouse anti-TIM-3 antibody competitive binding in a fluorescent activated cell sorting assay.

A representative graph showing the results of a competitive FACS-based assay comparing binding between parental mouse anti-TIM-3 antibody and 4 humanized anti-TIM-3 antibodies (ABTIM3-hum01, ABTIM3-hum04, ABTIM3-hum07 and ABTIM3hum08) and control hIgG4 is shown in fig. 7.

The results of multiple Biacore binding assays using surface plasmon resonance for a panel of humanized anti-TIM-3 antibodies are summarized in Table 1. 9.

Table 9. K _D values in Biacore against a panel of anti-TIM-3 antibodies.

All of the humanized mAbs tested have relatively the same affinity as all others and the parental mouse anti-TIM-3 antibody ranging from K _D 0.1-0.5 nM.

6.3. Binding to TIM-3 Expressing Cells

Humanized anti-TIM-3 antibodies were analyzed for binding to TIM-3 expressing cells using fluorescent activated cell sorting and Biacore assay as described in Example 1. FIG. 8A, binding of various humanized anti-TIM-3 antibodies to cells transfected with human TIM-3 was measured using flow cytometry. ABTIM3 was used as a positive control. Negative controls include hIgG4, goat anti-human, and secondary goat anti-mouse Ab with FITC. The results of the flow cytometric competition analysis were used to determine the dissociation constant (KD) of cells expressing human TIM-3, as shown in Table 1. 10 below.

Table 10. K _D values for binding to cells expressing huTIM-3.

Antibody K _D (nM) i : /i/ • < /7

A competitive binding assay was also performed to evaluate the binding of humanized anti-TIM-3 antibodies, ABTIM3-hum03 and ABTIM3-hum11, to cells expressing human TIM-3 in the presence of the mouse parental ABTIM3 antibody. As shown in FIG. 8B, humanized anti-TIM-3 antibodies competed with ABTIM3.

The K _D values for the two humanized anti-TIM-3 antibodies against the recombinant TIM-3-Ig fusion proteins were analyzed using surface plasmon resonance in the Biacore assay, as shown in Table 1. eleven.

- 154 040365

Table 11 Biacore KD values for TIM-3-Ig.

Cynomolgus TIM-3/Fc huTIM3/his mTIM-3/his Rat TIM-3/Fc ABTIM3- hum 03 K _D (M) 1.04E-09 1.24E-10 K _d (M) 3.89E-09 1.84E-10 5.10E-08 K _d (M) 3.08E-09 7.58E-11 Mean K _D (M) 2.67E-09 1.28E-10 ABTIM3- humi 1 K _d (M) 1.24E-09 1.55E-10 K _d (M) 3.14E-09 2.26E-10 K _d (M) 5.04E-09 1.09E-10 2.97E-07 Mean K _D (M) 3.14E-09 1.63E-10

These results indicate that humanized anti-TIM-3 antibodies have similar binding affinity to human and cynomolgus monkey proteins. The humanized anti-TIM-3 antibodies showed a weak binding affinity for rat TIM-3/Fc protein, on the order of 1/1000 of that for huTIM-3/Fc.

Example 7 X-Ray Crystal Structure of Human TIM-3/Fab ABTIM3hum21 Complex.

The crystal structure of human TIM-3 (IgV domain, SEQ ID NO: 220, Table 12) bound to the Fab fragment of the humanized anti-TIM-3 antibody ABTIM3-hum21 (SEQ ID NO: 221 and 222, Table 12) was determined. As defined below, TIM-3 was co-expressed with MGB220 Fab in mammalian cells to give a purified complex. Protein crystallography was then used to obtain atomic resolution data for TIM-3 bound to Fab ABTIM3-hum21 to determine the epitope. ABTIM3-hum21, a humanized antibody from a mouse parent antibody, contains an IgG1 framework region and a heavy chain variable region of SEQ ID NO: 84 and a light chain variable region of SEQ ID NO: 88. ABTIM3-hum21 differs by only one amino acid in the heavy chain CDR2 from the humanized anti-TIM antibodies described herein and this different amino acid (Gln55) is far (>6A) from the epitope and thus cannot alter antigen binding, indicating that the obtained crystal structure results are applicable to other humanized antibodies described in the present description.

7.1 Protein production

The TIM-3 and Fab ABTIM3-hum21 sequences obtained for crystallography are shown in Table 1. 12. The TIM-3 construct contains residues 22-130 (underlined) of human TIM-3 (UniProt ID Q8TDQ0, SEQ ID NO: 129) along with N- and C-terminal residues from a recombinant expression vector (shown in lower case letters, SEQ ID NO: : 130). The N-terminal signal sequence from the mouse IgG kappa light chain was used for secretion after TIM-3 expression and was cleaved off during expression, leaving the intact N-terminus of TIM-3. The C-terminus of TIM-3 was fused to a 6x His tag (SEQ ID NO: 133) for purification. For Fab ABTIM3-hum21, the heavy (SEQ ID NO: 131) and light (SEQ ID NO: 132) chain sequences are shown.

- 155 040365

Table 12. Amino acid sequences used by ________ to determine the crystal structure. _______

Design Amino acid sequence SEQ ID NO TIM-3 human MFSHLPFDCVLLLLLLLTRSSEVEYRAEVGQNAYLPC FYTPAAPGNLVPVCWGKGACPVFEECGNWLRTDERDVN YWTSRYWLNGDFRKGDVSLTIENVTLADSGIYCCRIQI PGIMNDEKFNLKLVIKPAKVTPAP TRQRD FTAAFPRML (Q8TDQ0) TTRGHGPAETQTLGSLPDINLTQISTLANELRDSRLAN 129 expressing DLRDS GATIRIGIYIGAGICAGLALALIFGALIFKWYS HSKEKIQNLSLISLANLPPSGLANAVAEGIRSEENIYT IEENVYEVEEPNEYYCYVSSRQPSQPLGCRFAMP metdtlllwvlllwvpgstgSEVEYRAEVGQNAYLPCF i design YTPAAPGNLVPVCWGKGACPVFECCGNWLRTDERDVNY η ο η for TIM-3 WTSRYWLNGDFRKGDVSLTIENVTLADSGIYCCRIQIP J. and and human GIMNDEKFNLKLVIKhhhhhh heavy chain Fab ABTIM3- QVQLVQS GAEVKKPGASVKVS CKAS GYT FT S YNMHWVR QAPGQGLEWIGDIYPGQGDTSYNQKFKGRATMTADKST STVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLV 131 hum21 light chain Fab ABTIM3- TVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVP SSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQ WYQQKPGQPPKLLIYAASNVESGVPDRFSGSTKTGTDGFT LTISSLQAEDQVEKVES 132 hum21 AAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQW KVflHKLQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGL S S PVTKS FNRGEC

TIM-3 was co-expressed with ABTIM3-hum21 Fab in Expi293® cells to generate a complex for crystallography. In detail, 0.3 mg of TIM-3 encoding plasmid was mixed with 0.15 mg of ABTIM3-hum21 Fab heavy chain encoding plasmid and 0.15 mg of ABTIM3-hum21 Fab light chain encoding plasmid was diluted to 30 ml with Opti medium -MEM® I (Life Technologies) and incubated with 1.5 mg polyethyleneimine (Polysciences) in 30 ml of the same medium for 30 minutes. The mixture was then added to 0.6 L of Expi293® cells grown in suspension in Expi293® expression medium (Life Technologies) at 2 million cells/ml at 37° C. with 8% CO2 for transfection. After 5 days, the medium containing the TIM-3/Fab ABTIM3-HUM21 complex was collected by centrifugation. 5 ml Ni-NTA resin was added to the medium and stirring was continued at 4°C overnight. On the following knocks, the beads were loaded onto a gel filtration column and washed with 25 mM Hepes pH 7.4, 150 mM NaCl (HBS) supplemented with 20 mM imidazole. The complex was eluted with 3 column volumes (CV) of HBS with 500 mM imidazole and then dialyzed in HBS at 4° C. overnight. The following day, the complex was incubated with 1/10 (v/v) PNGase F (purified in situ) at 37° C. overnight to eliminate N-linked glycosylation. After deglycosylation, the mixture was coupled back to 5 ml of Ni-NTA resin, washed with HBS to remove PNGase F, and eluted with HBS with 500 mM imidazole. The eluate was then concentrated and applied to a HiLoad 16/600 Superdex 75 PG size-exclusion column (GE Healthcare) equilibrated with HBS. Peak fractions containing purified TIM-3/Fab ABTIM3-hum21 complex were analyzed by SDS-PAGE, pooled and concentrated for crystallization.

7.2. Crystallization and structure determination

The TIM-3/Fab ABTIM3-hum21 complex was concentrated to 12.5 mg/ml, centrifuged at 20,000 g for 10 min, and sieved to crystallize. Data acquisition crystals were grown by hanging drop vapor diffusion at 20°C. More specifically, 0.1 µl of the TIM-3/Fab ABTIM3hum21 complex was mixed with 0.1 µl of a reservoir solution containing 0.04 M potassium phosphate monobasic, 16% (w/v) PEG 8000 and 20% (v/v .) glycerin. The drop was then equilibrated against 45 µl of the same reservoir solution. Before data collection, the crystals were quickly frozen in liquid nitrogen.

The diffraction data were acquired at the 17-ID beamline at Advanced Photon Source (Argonne National Laboratory, USA) and processed using Autoproc (version 1.1.5, Global Phasing, LTD). Data for TIM-3/Fab ABTIM3-hum21 was processed to 2.0 A in space group P21 with cell sizes a=84.3 A, b=93.0 A, c=85.3 A, alpha=90°, beta =114° and gamma=90°. The structure of the complex was determined by the molecular displacement method using Phaser (version 2.5.5, McCoy et al., (2007) J. Appl. Cryst. 40:658-674) with the structures of mouse TIM-3 (PDB ID: 3KAA) and Fab ( own structure) as search models. The final model was designed in COOT (version 0.6 pre, Emsley & Cow

- 156 040365 tan (2004) Acta Cryst. D60:2126-2132) and refined using Phenix (version 1.9, Afonine et al., (2012) Acta Cryst. D68:352-367). The R _work and R _free values were 17.5% and 22.1%, respectively; and the root mean square (rms) deviations of bond lengths and bond angles are 0.007 A and 1.1°, respectively.

An epitope was defined as TIM-3 residues that contain atoms within 5 A of any atom in the Fab ABTIM3-hum21 identified by CONTACT in the CCP4 software package (version 6.2.0, Winn et al., (2011) Acta. Cryst . D67:235-242) and are given in Table. 13. In the asymmetric element (the smallest unique element in the crystal) there are 2 copies of the TIM-3/Fab ABTIM3-hum21 complexes, only those antibody-contacting residues that are common in both copies are listed as epitope residues.

7.3. ABTIM3-hum21 epitope on TIM-3

The crystal structure of the TIM-3/Fab ABTIM3-hum21 complex was used to identify the ABTIM3-hum21 epitope on TIM-3. The interaction surface on TIM-3 with ABTIM3-hum21 was formed by several contiguous and discontinuous (i.e., non-contiguous) sequences, namely, residues Val24, Glu25, Tyr26, Phe39, Tyr40, Thr41, Gly56, Ala57, Cys58, Pro59, Val60 , Phe61, Ser105, Gly106, Ile107, Asn119, Asp120, Glu121, Lys122, Phe123, Asn124, Leu125, Lys126, Leu127, and Val128, as detailed in Table 1. 13. These residues form an exemplary three-dimensional conformational epitope that is recognized by ABTIM3-hum21 (FIG. 9).

Table 13. Interactions between human TIM-3 and ABTIM3-hum21. TIM-3 residues are numbered as in UniProt entry Q8TDQ0 (SEQ ID NO: 219). Antibody residues are numbered based on their linear amino acid sequence (SEQ ID NOs: 221 and 222) and the corresponding chains are designated (H for heavy chain, L for light chain). The TIM-3 residues shown here have at least one atom within 5 A of the ABTIM3-hum21 atom to account for potential water-mediated interactions.

TIM-3 ABTIM3-hum21 Amino acid Number Amino acid Number Chain Vai 24 Ala 102 Η asp 98 L Glu 25 Tyr 31 L Arg 96 L Tugh 26 Tyr 31 L Phe 39 Ser 31 H Tyr 52 H Tugh 40 Ser 31 H Thr 28 H Thr 41 Thr 28 H gly 56 Thr 34 L Ala 57 Phe 103 H Thr 34 L Asn 57 L Tyr 53 L Ala 54 L Cys 58 Tyr 53 L Asn 57 L Pro 59 Asn 57 L Tyr 53 L Vai 60 Asn 57 L Tyr 53 L

- 157 040365

Vai 58 L Ser 60 L Glu 59 L Phe 61 Ser 60 L Ser 105 Tyr 32 L gly 106 Tyr 31 L Tyr 32 L He 107 Phe 103 H Thr 34 L Tyr 31 L Leu 36 L Asn 119 Ser 60 L asp 120 Tyr 32 H Glu 121 Tyr 32 H Thr 28 H Lys 122 Tyr 32 H gly 100 H Tyr 53 L Glu 59 L Phe 123 gly 100 H gly 101 H Tyr 32 H Asn 124 Phe 103 H Ala 102 H Pro 104 H Tyr 53 L Leu 125 Ala 102 H Lys 126 Ala 102 H Tyr 31 L Leu 36 L Ser 95 L Lys 97 L Leu 127 Tyr 31 L Vai 128 Tyr 31 L Tyr 32 L

7.4. ABTIM3-hum21 vs. TIM-3 ligands

The identification of a TIM-3 epitope recognized by an anti-TIM-3 antibody indicates that the binding of some of the TIM-3 ligands may be impaired by antibody binding. Known TIM-3 ligands include CEACAM-1, phosphatidylserine (PtdSer), HMGB1, and galectin-9 (Gal-9).

With regard to CEACAM-1, a recent study showed that CEACAM-1 is the ligand for TIM-3 required for its ability to mediate T-cell inhibition, and this interaction has a key role in the regulation of autoimmunity and antitumor immunity (Huang et al., ( 2014) Nature. The same study also identified, both biochemically and structurally, key TIM-3 amino acid residues mediating its binding to CEACAM-1 (FIG. 10A). The ABTIM3-hum21 epitope on TIM-3 overlaps with these CEACAM-1 binding residues (FIG. 10A), including C58, N119 and K122. For example, K122 forms a hydrogen bond with N42 of CEACAM-1, however, it is completely blocked by ABTIM3-hum21 (Fig. 10B). The superposition of the crystal structures obtained for the TIM-3/Fab ABTIM3-hum21 and TIM-3/CEACAM-I complexes (PDB ID: 4QYC) resulted in a pronounced collision between ABTIM3-hum21 and CEACAM-1 (Fig. 10C). Taken together, these data indicate that ABTIM3-hum21 disrupts CEACAM-1 binding.

With respect to PtdSer, the FG loop and the CC' loop of TIM-3 form a pocket (often referred to as the metal ion dependent ligand binding site (MILIBS)), which has been shown by crystal structure to bind Ca ²⁺ and PtdSer simultaneously ( DeKruyff, et al., (2010) J Immunol 184(4):1918-1930). This binding is believed to help TIM-3 expressing cells bind to and penetrate the membranes of apoptotic cells (which expose PtdSer) for uptake. The TIM-3/Fab crystal structure of ABTIM3-hum21 indicates that ABTIM3-hum21 binds the PtdSer-binding loops of the human TIM-3 IgV domain; and the angle of attack of the antibody indicates that it interferes with PtdSer mediated penetration of TIM-3 across the membrane (FIG. 11).

With respect to HMGB1, it has been reported to interact with TIM-3 to help tumor-associated dendritic cells suppress nucleic acid-mediated immune response (Chi

- 158 040365 ba et al., (2012) Nat. Immunol. 13(9):832-842). The amino acid residue at position 62 of TIM-3 (Q in mouse TIM-3, E in human TIM-3) has been shown to be important for the binding of mouse HMGB1 to mouse TIM3. E62 is not present in the ABTIM3-hum21 epitope, although it is very close to the two epitope residues V60 and F61, so there is a possibility that ABTIM3-hum21 may block HMGB1 binding depending on the angle of attack of HMGB1 on TIM-3.

With respect to Gal-9, it has been shown to bind mouse TIM-3, negatively regulating the Th1 immune response (Zhu et al., (2005) Nat. Immunol, 6(12):1245-1252). However, it has also been reported that human TIM-3 on T cells does not act as a receptor for Gal-9 (Leitner et al., (2013) PLoS Pathog. 9(3):e1003253). For the crystal structure of human TIM-3/Fab ABTIM3-hum21, half of the putative Gal-9 binding site in mouse TIM-3 is not conserved in human TIM-3 (N74 and N90 in mouse TIM-3 become D74 and R89 in human TIM-3 ), i.e. this half region in human TIM-3 is unable to bind to Gal-9. The remaining half region (N33 and N99 in human TIM-3) is conserved but far from the ABTIM3-hum21 epitope on TIM-3 (FIG. 9A). Thus, even if Gal-9 is a human TIM-3 ligand, ABTIM3-hum21 will not interfere with Gal-9 binding to human TIM-3.

7.5. Experimental conditions for hydrogen-deuterium exchange

HDx/MS experiments were performed using methods similar to those described in the literature (Chalmers et al., (2006) Anal. Chem. 78(4):1005-1014). Experiments were performed on the Waters HDx/MS platform, which includes a LEAP autosampler, nanoACQUITY UPLC, and a Synapt G2 mass spectrometer. The deuterium buffer used to label the human TIM-3 protein backbone (a.k. 22135; SEQ ID NO: 139) was 25 mM HEPES, 150 mM NaCl, 5 mM CaCl ₂ pH7.4 with deuterium; the total percentage of deuterium in solution was 94.2%. For deuterium labeling experiments with human TIM-3 (a.a. 22-135) in the absence of antibody 300 pmol human TIM-3 (a.a. 22-135), volume 7.7 µl, diluted with 100 µl deuterium buffer in a chilled tube and incubated for 15 min on a shaker at 4°C. The labeling reaction was then quenched with 100 μl of chilled quench buffer at 2° C. for 5 min, followed by injection into an LC-MS system for automatic pepsin digestion and peptide analysis.

For deuterium labeling experiments with human TIM-3 (a.k. 22-135) in the presence of antibodies, 400 pmol of ABTIM3-hum03 or ABTIM3-hum11 were first immobilized on Thermo Protein G Plus beads and cross-linked using disuccinimidyl suberate (DSS). For labeling experiments, antibody beads (containing 400 pmol of antibody) were incubated with 300 pmol of human TIM-3 (a.k. 22-135) for 25 min at 4°C. After 25 min, the beads were washed with 200 µl of HEPES buffer. Then 200 μl of chilled deuterium buffer (87.5% deuterium) was added and the complex was incubated for 15 min at 4°C. After 15 min, the deuterium buffer was removed by centrifugation and the labeling reaction was quenched with 200 µl of chilled quench buffer on ice for 4 min. After centrifuging the sample for 30 s in a centrifuge, the quenched solution was injected into the LC-MS system for automatic pepsin digestion and peptide analysis.

All deuterium exchange experiments were quenched using 1 M TCEP and 6 M urea (pH 2.6). After quenching, the exchanged antigen was sequentially digested with pepsin using a Poroszyme Immobilized Pepsin column (2.1 x 30 mm) at 12° C. and then captured on a Waters Vanguard HSS T3 capture column. The peptides were eluted from the capture column and separated on a Waters BEN C18 1x100 mm column (maintained at 1°C) at a flow rate of 40 μl/min using a binary gradient over 8.4 min from 2 to 35% B (mobile phase A was 99.9% water and 0.1% formic acid; mobile phase B was 99.9% acetonitrile and 0.1% formic acid).

7.6 Results of hydrogen-deuterium exchange

For human TIM-3, 93% of the sequence was monitored by deuterium exchange as shown in FIG. 18. In this figure, each bar corresponds to a peptide that was monitored in all deuterium exchange experiments. For differential experiments between the bound and unbound state of antibodies, it is informative to investigate the difference in deuterium uptake between the two states. In FIG. 19, a negative value indicates that the TIM-3-antibody complex undergoes less deuterium capture relative to TIM-3 alone. The decrease in deuterium capture may be due to protection of the region from exchanged deuterium or stabilization of the hydrogen bond network. Conversely, a positive value indicates that the complex undergoes higher deuterium uptake relative to TIM-3 alone. The increase in deuterium uptake may be due to destabilization of hydrogen bond networks (i.e., localized protein unfolding).

ABTIM3-hum03 has identical CDRs to ABTIM3-hum11, except that ABTIM3hum03 has a glutamine at position 55 of HCDR2 while ABTIM3-hum11 has an asparagine at position 55 of HCDR2. ABTIM3-hum03 has the same CDRs as ABTIM3-hum21. These antibodies would be expected to share the same epitope on TIM-3. From FIG. 19 shows that ABTIM3-hum03 and

- 159 040365

ABTIM3-hum11 show the same protection profile, which is consistent with the fact that these two antibodies share the same epitope. A more detailed study of Fig. 19 demonstrates that when TIM-3 is complexed with either of the two antibodies, many regions of TIM-3 are significantly protected, generally defined as protection less than or equal to -0.5 Da (Houde et al. (2010) J Pharma Sci 100(6):2071-2086). The observation of broad protection indicates that binding of either of the two antibodies to the TIM-3 antigen causes broad stabilization based on hydrogen bond networks in TIM-3. This broad protection is in addition to the protection that results from solvent shielding of the epitope at the antibody-antigen interface. Given the significant level of broad protection, it is useful to rank the order of the most protected regions of TIM-3 when binding an antibody to identify regions likely to be involved in an epitope. TIM-3 regions that are most protected when ABTIM3-hum03 or ABTIM3-hum 11 bind include regions 23-25 (EVE), 41-61 (TPAAPGNLVPVCWGKGACPVF, SEQ ID NO: 140), 73-77 (RDVNY, SEQ ID NO : 141), and 121-127 (EKFNLKL, SEQ ID NO: 142). Comparison of these protected regions with X-ray crystallography data summarized in Table 1. 13 shows a stable match indicating that the epitope determined by the x-ray crystal structure is present in solution.

Example 8 TIM-3 expression in cancer.

TIM-3 is expressed in various cancers. In this example, several different assay methods were used to identify TIM-3 expressing cancers in which therapeutic benefit could be achieved using an anti-TIM-3 antibody.

8.1. Immunohistochemical staining of tumors

ABTIM-3 has been used to stain various tumor tissues. TIM-3 expression in tumors has been identified in esophageal squamous cell carcinoma, primary and metastatic renal cell carcinoma, colorectal cancer, and leukemic stem cells in AML.

8.2. Expression analysis in the TCGA and ICGC databases

In general, TIM-3 expression was compared with The Cancer Genome Atlas (TCGA) database and the International Cancer Genome Consortium (ICGC) database. The following malignancies have been identified as having the highest expression of TIM-3: diffuse large B cell lymphoma (DLBCL), clear cell renal cell carcinoma (KIRC), glioblastoma multiforme (GBM), nasopharyngeal carcinoma (NPC), lung adenocarcinoma (LUAD), papillary cell carcinoma kidney (KIRP), mesothelioma (MESO), acute myeloid leukemia (AML), and triple negative (TN) immunomodulatory (IM) subtype breast cancer (FIG. 12).

Next identified malignant tumors, which were characterized by high expression of TIM-3 in conjunction with high expression of other markers of immune cells. Other immune cell markers include: T cell marker CD3e, T regulatory cell marker FoxP3, natural killer marker NKp30, macrophage marker CD68, and dendritic cell marker CD11c. As shown in FIG. 12, cancers with high expression of TIM-3 and other immune cell markers have been identified. High expression was quantified using 3rd quartile (or 25% highest) expressing tumors among more than 34,000 cases. For TIM-3 and CD3e, the major diseases were large B cell lymphoma (DLBCL), nasopharyngeal carcinoma (NPC), and clear cell renal cell carcinoma (KIRC). For TIM-3 and FoxP3, the main diseases were diffuse large B-cell lymphoma (DLBCL), nasopharyngeal carcinoma (NPC), and lung adenocarcinoma (LUAD). For TIM-3 and NKp30, the main indications were diffuse large B cell lymphoma (DLBCL), nasopharyngeal carcinoma (NPC) and acute myeloid leukemia (AML). For TIM-3 and CD68, the main diseases were diffuse large B-cell lymphoma (DLBCL), renal clear cell carcinoma (KIRC), and renal papillary cell carcinoma (KIRP). For TIM-3 and CDllc, the main diseases were diffuse large B-cell lymphoma (DLBCL), mesothelioma (MESO) (although only a small sample was evaluated), and renal papillary cell carcinoma (KIRP).

The correlation between TIM-3 or PD-1 and T-cell-associated or macrophage-associated markers was also compared in the TCGA database. The assay demonstrated a correlation between TIM-3 expression with both T-cell-associated markers (eg, ZAP70, CD3D, CD3G, CD8B, GZMH, GZMK, and ITK) and macrophage-associated markers (eg, LILRB4, MRC1, MSR1 , SIGLEC1, TREM2, CD163, ITGAX, and ITGAM), but TIM-3 expression is more associated with macrophage markers, especially inhibitory receptors on macrophages (eg, LILRB4). The expression profile of macrophages, eg, macrophage-associated markers (eg, LILRB4, MRC1, MSR1, SIGLEC1, TREM2, CD163, ITGAX, and ITGAM), was determined for various cancers, and FIG. 13 they are organized by the highest expressing macrophage profile tumors. Cancers with a high expression of the macrophage profile are the same diseases as in the case of high expression of TIM-3.

- 160 040365

Example 9 Patient selection based on PDL1/CD8/IFN-y status.

For each of several types of cancer, samples from multiple patients were examined for PDL1/CD8/IFN-y status. Each sample was classified as: triple negative for PDL1/CD8/IFN-y, single or double positive for these markers, or triple positive for these markers. In FIG. 14 shows that in this experiment, in the patient population, the following types of cancer are often triple positive for PDL1/CD8/IFN-y: lung cancer (squamous cell), lung cancer (adenocarcinoma), head and neck cancer, cervical cancer (squamous cell) , stomach cancer, thyroid cancer, melanoma and nasopharyngeal cancer. Patients having these types of cancers are good candidates for anti-PD-1 antibody therapy and combination therapies as described herein. The likelihood of successful treatment can be further enhanced by determining which patients are PDL1/CD8/IFN-y triple positive and treating the triple positive patients with anti-TIM-3 antibodies, alone or in combination with one or more immunomodulators (eg, a PD inhibitor). -1 or PD-L1 inhibitor), and/or combination therapies as described herein.

In FIG. 15 shows that the following types of cancer are rarely triple positive for PDL1/CD8/IFN-y in the patient population: ER+ breast cancer and pancreatic cancer. Remarkably, even in cancers that are not normally PDL1/CD8/IFN-y positive, it is possible to increase the likelihood of successful treatment by determining which patients are PDL1/CD8/IFN-y triple positive and treating the triple positive patients with antibodies against TIM-3, alone or in combination with one or more immunomodulators (eg, PD-1 inhibitor or PD-L1 inhibitor) and/or combination therapies as described herein.

In FIG. 16 shows the proportion of breast cancer patients who are triple positive for PDL1/CD8/IFN-y. Considering breast cancer in general, the proportion of triple positive patients is quite low. However, if one focuses only on IM-TN breast cancer, one can see that a significantly higher percentage of patients are triple positive for PDL1/CD8/IFN-y. IM-TN breast cancer is particularly difficult to treat with conventional therapies. The discovery that IM-TN breast cancer is often triple positive for PDL1/CD8/IFN-y provides new avenues for therapy of this malignancy with anti-TIM-3 antibodies, alone or in combination with one or more immunomodulators (eg, a PD-1 inhibitor). or PD-L1 inhibitor) and/or combination therapies as described herein.

In FIG. 17 shows the proportion of patients with colon cancer who are triple positive for PDL1/CD8/IFN-y. Considering colon cancer as a whole, the proportion of triple positive patients is quite low. However, if one focuses only on breast cancers with high MSI (high microsatellite instability), it can be seen that a significantly higher percentage of patients are triple positive for PDL1/CD8/IFN-y. MSI levels can be analyzed using, for example, commercially available PCR-based methods.

Gastric cancer samples were examined for PDL1/CD8/IFN-y levels (data not shown). It was found that in high MSI gastric cancer or EBV+ gastric cancer, approximately 49% were positive for PDL1, and a high proportion of PDL1 positive cells were triple positive for PDL1/CD8/IFN-y. It was also found that a proportion of PDL1 positive cells and PDL1/CD8/IFN-y positive cells were also positive for PIK3CA. These data indicate that these cancers can be treated with an anti-TIM-3 antibody, alone or in combination with one or more immunomodulators (e.g., a PD-1 inhibitor or a PD-L1 inhibitor), optionally in combination with a therapeutic agent that targets PIK3.

High MSI CRC samples were tested for a combination of markers (data not shown). In high MSI CRC samples, a high proportion of PDL1/CD8/IFN-y samples were also found to be positive for LAG-3, PD-1 (also referred to as PDCD1), RNF43 and BRAF. These data indicate that these cancers can be treated with an anti-TIM-3 antibody, optionally in combination with a therapeutic agent that targets one or more of LAG-3, PDCD1, RNF43 and BRAF.

Squamous cell lung cancer was examined for a combination of markers (data not shown). In squamous cell lung cancer samples, a high proportion of PDL1/CD8/IFN-y positive samples were also found to be LAG-3 positive. These data indicate that these cancers can be treated with an anti-TIM-3 antibody, optionally in combination with a therapeutic agent that targets LAG-3, such as an anti-LAG-3 antibody.

Papillary thyroid cancer was investigated for a combination of markers including the BRAF V600E mutation (data not shown). It was found that a high proportion of cancer samples

- 161 040365 thyroid glands that are positive for PDL1 are also positive for V600E BRAF. These data indicate that these cancers can be treated with an anti-TIM-3 antibody, alone or in combination with one or more immunomodulators (e.g., a PD-1 inhibitor or a PD-L1 inhibitor), optionally in combination with a therapeutic agent that targets on BRAF.

Example 10 Selection of Patients Based on PD-L1 Status.

In order to provide extensive cancer research into PD-1/PD-L1 based therapies, the present inventors evaluated PD-L1 expression at both protein and mRNA levels in human cancers, including tumors of both the lung and and liver.

PD-L1 protein expression was assessed in a set of formalin-fixed paraffin-embedded lung non-small cell adenocarcinoma (ASA) (NSCLC), NSCLC squamous cell carcinoma (SCC) and hepatocellular carcinoma (HCC) tumors using immunohistochemistry (IHC). PD-L1 expression was assessed semi-quantitatively using a manual histo-score (H-score) methodology based on staining intensity and percentage of positive tumor cells. In the IHC assay conducted by the present inventors, PD-L1 positivity (PD-L1+) was defined as an H-score >20. In parallel, PD-L1 mRNA expression data from The Cancer Genome Atlas (TCGA) were examined in these indications (503 NSCLC ACA, 489 NSCLC SCC and 191 HCC) and analyzed by comparing expression in corresponding normal tissues from TCGA.

Using RNAseq analysis, data were calculated as Iog2 (RPKM+0.1) after RSEM normalization using the OmicSoft RNASeq system for oncology from TCGA. PD-L1 expression is elevated in ACA and SCC NSCLC relative to HCC. By overlaying distributions and comparing expression levels among all TCGA indications, the present inventors ranked the overexpression profiles for PD-L1 and found that the TCGA HCC group had significantly reduced levels of PD-L1 mRNA with a mean level of -0.8 compared to 1. 3 for ACA and 1.5 for SCC, which corresponds to a change from the average expression level by more than 2 times. Using RNAseq, the present inventors identified 50% of NSCLC adenocarcinoma cases, 54% of NSCLC squamous cell carcinoma cases, and 6% of HCC as high-expressing PD-L1.

Expression of PD-L1 protein in tumor cells was measured in 45 lung adenocarcinoma (ACA), 47 lung squamous cell carcinoma (SCC) and 36 hepatocellular carcinoma (HCC) samples. 16/45 (35.6%) cases of lung ACA, 21/47 (44.7%) cases of lung SCC were positive for PD-L1. In contrast, PD-L1 positivity was observed in only 2/36 (5.6%) HCC samples.

In general, using IHC and RNAseq analysis in large and independent sets of NSCLC and human HCC samples, the present inventors found that PD-L1 expression is more upregulated in NSCLC than in HCC. In the case of NSCLC, the data between adenocarcinoma and squamous cell carcinoma were comparable. Importantly, among the large number of samples (128 for IHC and 1183 for RNAseq) for the 3 diseases, there is a very high agreement between protein and mRNA based assays. Thus, the data obtained by the present inventors establish the basis for large-scale mRNA-based data collection in TCGA for diseases and patient segments in which PD-I/PD-LI- and/or TIM3-based immune therapy responses can be enhanced. .

Example 11 Competitive assays indicate that humanized anti-TIM3 antibodies bind to similar epitopes.

As described above, the TIM-3 epitope recognized by ABTIM3-hum21 was determined using X-ray crystallography based studies. ABTIM3-hum21 differs by only one amino acid in the heavy chain CDR2 from other humanized anti-TIM3 antibodies described herein, and this differing amino acid (Gln55) is located far (>6A) from the epitope and thus cannot be expected to alter binding antigen. Two different competitive assays were performed to compare epitope binding between ABTIM3-hum21 and two other humanized anti-TIM3 antibodies, ABTIM3-hum03 and ABTIM3-hum11. The results of both competition assays showed that both ABTIM3-hum04 and ABTIM3-hum11 effectively compete with ABTIM3-hum03 for binding to TIM3, thus demonstrating that ABTIM3-hum03 and ABTIM3-hum11 also bind to an epitope similar to ABTIM3-hum21 , for example an epitope as described herein.

11.1. Flow cytometric competitive analysis

The K _{D of} ABTMI3-hum21 was determined by labeling ABTIM3-hum21 with phycoerythrin, incubation with hTIM-3 expressing 300.19 cells, and a binding curve was generated to determine a KD of 2.15.

Titrated concentrations of unlabeled hIgG1 (isotype control), ABTIM3-hum21 (poly

- 162 040365 living control), ABTIM3-hum11 or ABTIM3-hum03 were mixed with ABTIM3-hum21 at its KD and incubated with hTIM-3 expressing 300.19 cells at 4°C for 3 h. Cells were washed twice and analyzed on an LSRFortessa flow cytometer . Data was analyzed in FlowJo and MFI (PE) values were plotted and plotted using GraphPad (Prism) software. The experiment was carried out twice.

Competitive assay results demonstrate that both ABTIM3-hum11 and ABTIM3-hum03 (but not the isotype control) competed with ABTIM3-hum21 for binding to human TIM3 expressed on 300.19 cells (FIG. 20). From binding curves, KDs were calculated for ABTIM3-hum11 and ABTIM3-hum03; the calculated KD for ABTIM3-hum11 was 2.276 nM and the calculated KD for ABTIM3-hum03 was 2.413 nM. These results demonstrate that ABTIM3-hum11 and ABTIM3hum03 bind to a similar or the same epitope as ABTIM3-hum21. 11.2

Biacore Competitive Analysis

The hTIM-3/his antigen was captured using an immobilized anti-His antibody (RU10000) on a CM5 chip. The first antibody was injected until saturation (>90%) was reached. A second antibody was injected to assess whether a second binding event had occurred. The occurrence of a second binding event indicates that the two antibodies tested have different epitopes. The absence of a second binding event indicates that the two antibodies can recognize and bind to the same epitope. Control assays were performed such that the test antibody was assayed with an isotype control human IgG1 or the test antibody was assayed as first and second antibodies (eg, self-self cycle) to observe a baseline binding event. In table. 14 summarizes the Biacore cycles analyzed and shows which antibodies were used as first and second antibodies in each cycle.

Table 14. Summary of Biacore Competitive Analysis Runs.

Baseline detection and first and second binding events are recorded as RUs (resonance units) and this can be represented on a sensorgram. A typical sensorgram is shown in Fig. 21 where the binding event is shown after injection of the 1st antibody. After washing, a second antibody is injected and the second binding can be detected. Significant RU changes indicate a bind event. A summary of RU changes detected upon injection of the 1st and 2nd antibodies in the competitive Biacore binding assay is shown in Table 1. 15.

- 163 040365

Table 15 Summary of Biacore Competitive Analysis Results.

Injection of the 1st antibody Injection of the 2nd antibody huIgGl ABTIM3hum21 ΑΒΤΙΜ3- hum03 ΑΒΤΙΜ3humll huIgGl 0.27 3, 6 88.2 86.3 83.2 ABTIM3hum21 95, 85 4.5 6, 6 7, 6 8, 1 ABTIM3- hum03 93.33 4.5 6, 9 7.3 8.5 ABTIM3humll 93.48 3, 8 ΝΑ ¹ 5, 3 7.2

^3 sensogram value was not calculated due to an unknown fluid problem.

The results shown above demonstrate that injection of ABTIM3-hum21, ABTIM3hum03 and ABTIM3-huml 1 during injection of the first antibody results in a binding event. Injection of ABTIM3-hum21, ABTIM3-hum03, and ABTIM3-huml 1 as a second antibody after injection of a control human IgGl antibody results in a second binding event. However, injection of any of the anti-TIM3 antibodies tested here as the first and second antibodies did not result in a second binding event, demonstrating that each pair of l-ro and 2 antibodies tested competed for binding to the same epitope. TIM3. These results show that ABTIM3-hum21, ABTIM3-humO3 and ABTIM3-huml 1 bind to a similar or the same epitope on human TIM3.

Example 12 Pharmacokinetic properties of ABTIM3-huml 1.

Various pharmacokinetic properties of ABTIM3-huml 1 were evaluated in mouse and rat models. ABTIM3-humll was injected intravenously into mice at various doses: 1 mg/kg, 3 mg/kg and 10 mg/kg. Blood samples were obtained at various time points between 0 and 672 hours (0-28 days). 10 mg/kg ABTIM3-humll was injected intravenously into rats and blood samples were obtained at various time points ranging from 0-400 hours (0-16 days). The concentration of ABTIM3-huml 1 present in serum was determined (FIGS. 23A and 23B). The results showed that ABTIM3-huml 1 is stable in both mouse and rat serum. In table. 16 shows additional determined pharmacokinetic properties including half-life (T1/2), maximum serum concentration (Cmax), AUC up to last measurable concentration (AUClast), and AUC extrapolated to infinity (AUCinf).

Table 16 Summary of the pharmacokinetic properties of ABTIM3-huml 1.

View Dose (mg/kg) t h (h) Stax (mcg/ml) AUClast (h*mcg/ml) AUCinf (h*mcg/ml) Mouse 1 N 3 3 3 3 Average value 142.3 17.3 1507.8 1571.4 STD 96.9 0.7 337.9 439.5 3 N 3 3 3 3 Average value 266.1 37.2 4617.9 5369.0 STD 73.1 2.3 2109.8 2496.1 10 N 3 3 3 3 Average value 254.9 147.5 23906.5 28621.7 STD 39.2 13.2 4369.8 6314.1 Rat 10 N 3 3 3 3 Average value 400.8 243.4 26032.3 53767.1 STD 75.9 19.1 895.8 5362.6

In a toxicity study, three naive mice were given a single dose of 1 mg/kg, 3 mg/kg, or 10 mg/kg ABTIM3-huml 1 via intravenous injection. No adverse effects were observed after 28 days, indicating that the ABTIM3 antibody is tolerable in mouse models.

Inclusion as a link

All publications, patents and access numbers mentioned in the present description are incorporated herein by reference in their entirety, as if it were specifically or individually indicated that each individual publication or patent is incorporated by reference.

Equivalents

While specific embodiments of the compositions and methods described herein have been contemplated, the above description is illustrative, but not limiting. Many embodiments of the invention will become apparent to those skilled in the art upon examination.

- 164040365 of this description and the claims below. The full scope of the invention is to be determined by the claims, and their full scope equivalents and description, and such variations.

Claims (54)

1. An isolated antibody molecule capable of binding to the T-cell immunoglobulin domain and human mucin 3 (TIM-3) domain, containing: (a) a heavy chain variable region (VH) containing the amino acid sequence VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 10 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14; (b) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 4 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7 and the amino acid sequence of VLCDR3 SEQ ID NO: 8; (c) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 25 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14; (d) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 24 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7 and the amino acid sequence of VLCDR3 SEQ ID NO: 8; (e) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 31 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14; or (f) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 30 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7, and the amino acid sequence of VLCDR3 SEQ ID NO: 8. 2. An antibody molecule according to claim 1, wherein said antibody molecule is a monospecific antibody molecule. 3. An isolated bispecific antibody molecule capable of binding to the T-cell immunoglobulin domain and human mucin 3 (TIM-3) domain, containing: (a) a heavy chain variable region (VH) containing the amino acid sequence VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 10 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14; (b) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 4 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7 and the amino acid sequence of VLCDR3 SEQ ID NO: 8; (c) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 25 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14; (d) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 24 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and VL containing the amino acid sequence of VLCDR1 SEQ ID NO: - 165 040365 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7 and the amino acid sequence of VLCDR3 SEQ ID NO: 8; (e) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 31 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14; or (f) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 30 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7, and the amino acid sequence of VLCDR3 SEQ ID NO: 8. 4. The bispecific antibody molecule of claim 3, wherein said antibody molecule has a first binding specificity for TIM-3 and a second binding specificity for PD-1, LAG-3, CEACAM-1, CEACAM-5, PD-L1, or PD -L2. 5. An antibody molecule according to any one of claims 1 to 4, wherein said antibody molecule is a humanized antibody molecule. 6. An antibody molecule according to any one of claims 1 to 5, wherein said antibody molecule comprises an antigen-binding antibody fragment, or a half-antibody, or an antigen-binding fragment of a half-antibody. 7. An antibody molecule according to any one of claims 1 to 6, which is a Fab, F(ab') ₂ , Fv or a single chain Fv fragment (scFv). 8. An antibody molecule according to any one of claims 1 to 7, wherein the antibody molecule comprises a heavy chain constant region selected from IgG1, IgG2, IgG3 and IgG4 and/or a light chain constant region selected from kappa or lambda constant regions. 9. An antibody molecule according to any one of claims 1 to 8, which contains: VH containing an amino acid sequence selected from any of SEQ ID NOs: 1, 16, 26, 32, 36, 44, 48, 52, 60, 68, 72, 76, 80, 84, 92, or 100, or an amino acid sequence, at least 85% identical to any of SEQ ID NOs: 1, 16, 26, 32, 36, 44, 48, 52, 60, 68, 72, 76, 80, 84, 92, or 100, and/or VL containing an amino acid sequence selected from any of SEQ ID NOs: 2, 20, 40, 56, 64, 88, 96, or 104, or an amino acid sequence at least 85% identical to any of SEQ ID NOs: 2, 20 , 40, 56, 64, 88, 96 or 104. 10. An antibody molecule according to any one of claims 1 to 9, which contains: VH containing an amino acid sequence selected from any of SEQ ID NOs: 1, 16, 26, 32, 36, 44, 48, 52, 60, 68, 72, 76, 80, 84, 92, or 100, and/or A VL containing an amino acid sequence selected from any of SEQ ID NOs: 2, 20, 40, 56, 64, 88, 96, or 104. 11. An antibody molecule according to any one of claims 1 to 10, which contains: a heavy chain containing an amino acid sequence selected from any of SEQ ID NOs: 18, 121, 28, 34, 38, 116, 46, 50, 54, 62, 70, 74, 78, 82, 86, 94 or 102, and /or a light chain containing an amino acid sequence selected from any of SEQ ID NOs: 22, 42, 58, 66, 90, 98, or 106. 12. An antibody molecule according to any one of claims 1 to 10, which contains: (a) VH containing the amino acid sequence of SEQ ID NO: 1 and VL containing the amino acid sequence of SEQ ID NO: 2, (b) VH containing the amino acid sequence of SEQ ID NO: 16 and VL containing the amino acid sequence of SEQ ID NO: 20, (c) VH containing the amino acid sequence of SEQ ID NO: 26 and VL containing the amino acid sequence of SEQ ID NO: 20, (d) VH containing the amino acid sequence of SEQ ID NO: 32 and VL containing the amino acid sequence of SEQ ID NO: 20, (e) VH containing the amino acid sequence of SEQ ID NO: 36 and VL containing the amino acid sequence of SEQ ID NO: 40, (f) VH containing the amino acid sequence of SEQ ID NO: 44 and VL containing the amino acid sequence SEQ ID NO: 40, (g) VH containing the amino acid sequence of SEQ ID NO: 48 and VL containing the amino acid sequence of SEQ ID NO: 40, (h) VH containing the amino acid sequence of SEQ ID NO: 36 and a VL containing the amino acid sequence of SEQ ID NO: 20, (i) a VH containing the amino acid sequence of SEQ ID NO: 16 and a VL containing the amino acid sequence of SEQ ID NO: 40, - 166 040365 (j) VH containing the amino acid sequence of SEQ ID NO: 52 and VL containing the amino acid sequence of SEQ ID NO: 56, (k) VH containing the amino acid sequence of SEQ ID NO: 60 and VL containing the amino acid sequence of SEQ ID NO: 56, (l) VH containing the amino acid sequence of SEQ ID NO: 52 and VL containing the amino acid sequence of SEQ ID NO: 64, (m) VH containing the amino acid sequence of SEQ ID NO: 60 and VL containing the amino acid sequence of SEQ ID NO: 64, (n) VH containing the amino acid sequence of SEQ ID NO: 68, and VL containing the amino acid sequence of SEQ ID NO: 64, (o) VH containing the amino acid sequence of SEQ ID NO: 72, and VL, containing the amino acid sequence of SEQ ID NO: 64, (p) VH containing the amino acid sequence of SEQ ID NO: 76, and VL containing the amino acid sequence of SEQ ID NO: 56, (q) VH containing the amino acid sequence is SEQ ID NO: 80 and VL containing the amino acid sequence of SEQ ID NO: 56, (r) VH containing the amino acid sequence of SEQ ID NO: 68 and VL containing the amino acid sequence of SEQ ID NO: 56, (s) VH, containing the amino acid sequence of SEQ ID NO: 72, and VL containing the amino acid sequence of SEQ ID NO: 56, (t) VH containing the amino acid sequence of SEQ ID NO: 76, and VL containing the amino acid sequence of SEQ ID NO: 64, (u) VH containing the amino acid sequence of SEQ ID NO: 80 and VL containing the amino acid sequence of SEQ ID NO: 64, (v) VH containing the amino acid sequence of SEQ ID NO: 84 and VL containing the amino acid sequence of SEQ ID NO: 88, ( w) a VH containing the amino acid sequence of SEQ ID NO: 92 and a VL containing the amino acid sequence of SEQ ID NO: 96, or (x) a VH containing the amino acid sequence of SEQ ID NO: 100 and a VL containing the amino acid sequence ovality SEQ ID NO: 104. 13. An antibody molecule according to any one of claims 1 to 8, which contains: (a) a heavy chain containing the amino acid sequence of SEQ ID NO: 18 and a light chain containing the amino acid sequence of SEQ ID NO: 22, (b) a heavy chain containing the amino acid sequence of SEQ ID NO: 28 and a light chain containing the amino acid sequence SEQ ID NO: 22, (c) a heavy chain containing the amino acid sequence of SEQ ID NO: 34 and a light chain containing the amino acid sequence of SEQ ID NO: 22, (d) a heavy chain containing the amino acid sequence of SEQ ID NO: 38, and a light chain containing the amino acid sequence of SEQ ID NO: 42, (e) a heavy chain containing the amino acid sequence of SEQ ID NO: 46 and a light chain containing the amino acid sequence of SEQ ID NO: 42, (f) a heavy chain containing the amino acid sequence of SEQ ID NO: 50, and a light chain containing the amino acid sequence of SEQ ID NO: 42, (g) a heavy chain containing the amino acid sequence of SEQ ID NO: 116, and l a light chain containing the amino acid sequence of SEQ ID NO: 22, (h) a heavy chain containing the amino acid sequence of SEQ ID NO: 121 and a light chain containing the amino acid sequence of SEQ ID NO: 42, (i) a heavy chain containing the amino acid sequence of SEQ ID NO: 54, and a light chain containing the amino acid sequence of SEQ ID NO: 58, (j) a heavy chain containing the amino acid sequence of SEQ ID NO: 62, and a light chain containing the amino acid sequence of SEQ ID NO: 58, (k) a heavy a chain containing the amino acid sequence of SEQ ID NO: 54 and a light chain containing the amino acid sequence of SEQ ID NO: 66, (l) a heavy chain containing the amino acid sequence of SEQ ID NO: 62 and a light chain containing the amino acid sequence of SEQ ID NO: 66, (m) a heavy chain containing the amino acid sequence of SEQ ID NO: 70 and a light chain containing the amino acid sequence of SEQ ID NO: 66, (n) heavy th chain containing the amino acid sequence of SEQ ID NO: 74 and a light chain containing the amino acid sequence of SEQ ID NO: 66, (o) 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: : 58, (p) a heavy chain containing the amino acid sequence of SEQ ID NO: 82, and a light - 167 040365 a chain containing the amino acid sequence of SEQ ID NO: 58, (q) a heavy chain containing the amino acid sequence of SEQ ID NO: 70 and a light chain containing the amino acid sequence of SEQ ID NO: 58, (r) a heavy chain containing the amino acid sequence of SEQ ID NO: 74, and a light chain containing the amino acid sequence of SEQ ID NO: 58, (s) 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: 66, (t ) 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: 66, (u) a heavy chain containing the amino acid sequence of SEQ ID NO: 86 and a light chain containing the amino acid sequence of SEQ ID NO: 90, (v) a heavy chain containing the amino acid sequence of SEQ ID NO: 94 and a light chain containing the amino acid sequence of SEQ ID NO: 98, or (w) a heavy chain containing the amino acid sequence of SEQ ID NO: 102 and a light chain containing the amino acid sequence of SEQ ID NO: 106. 14. An antibody molecule according to any one of claims 1 to 13, which contains: (a) a human IgG4 heavy chain constant region with a mutation at position 228 according to EU numbering or at position 108 of SEQ ID NO: 108 or 110 and a kappa light chain constant region; (b) a human IgG4 heavy chain constant region with a serine to proline mutation at position 228 according to EU numbering or position 108 of SEQ ID NO: 108 or 110 and a kappa light chain constant region; (c) a human IgG1 heavy chain constant region with an asparagine to alanine mutation at position 297 according to EU numbering or at position 180 of SEQ ID NO: 112 and a kappa light chain constant region; (d) a human IgG1 heavy chain constant region with an aspartate to alanine mutation at position 265 according to EU numbering or position 148 of SEQ ID NO: 113 and a proline to alanine mutation at position 329 according to EU numbering or position 212 of SEQ ID NO: 113 and kappa light chain constant region; or (e) a human IgG1 heavy chain constant region with a leucine to alanine mutation at position 234 according to EU numbering or position 117 of SEQ ID NO: 114 and a leucine to alanine mutation at position 235 according to EU numbering or position 118 of SEQ ID NO: 114 and kappa light chain constant region. 15. An antibody molecule according to any one of claims 1-14, which has one, two, three or all of the following properties: (a) capable of binding to TIM-3 with a dissociation constant (KD) of less than about 0.5 nM; (b) is capable of reducing: (i) TIM-3 binding to phosphatidylserine (PtdSer), HMGB1, CEACAM-1, semaphorin 4-A, or a combination thereof; or (ii) PtdSer mediated membrane permeation of TIM-3; (c) capable of enhancing the response of antigen-specific T cells; or (d) binds the IgV domain of TIM-3. 16. An isolated antibody molecule that binds to the same epitope as, or an epitope that overlaps with, an epitope of a monoclonal antibody on the human T-cell immunoglobulin and mucin 3 (TIM-3) domain, wherein the monoclonal antibody contains: (a) a heavy chain variable region (VH) containing the amino acid sequence VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 10 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14; (b) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 4; and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7 and the amino acid sequence of VLCDR3 SEQ ID NO: 8; (c) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 25 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14; (d) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; - 168 040365 the amino acid sequence of VHCDR2 SEQ ID NO: 24 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7 and the amino acid sequence of VLCDR3 SEQ ID NO: 8; (e) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 31 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14; or (f) VH containing the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 30 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL containing the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7, and the amino acid sequence of VLCDR3 SEQ ID NO: 8, where (1) the antibody molecule binds to one, two, three, or all of: two residues , adjacent to the N-terminus of the A chain (Val24 and Glu25 in human TIM-3), the BC loop, the CC' loop, or the G chain of human TIM-3; and (2) the antibody molecule has one, two, three, four, five, or all of the following properties: (i) reduces PtdSer-dependent membrane permeation of TIM-3; (ii) reduces TIM-3 binding to one, two, or all of PtdSer, HMGB1, or CEACAM-1; (iii) competes with CEACAM-1 for binding to one, two, or all of TIM-3's Cys58, Asn119, and Lys122; (iv) reduces the formation of hydrogen bonds between Lys122 TIM-3 and Asn42 CEACAM-1; (v) competes with PtdSer for binding to the FG loop and the CC' loop of TIM-3; or (vi) competes with HMGB1 for binding to TIM-3 Glu62. 17. An antibody molecule according to claim 16, wherein the antibody molecule binds to one, two, three, four, five, six, seven, eight, or all of the following: (a) one, two, three, or all of: two residues adjacent to the N-terminus of the A chain (Val24 and Glu25 in human TIM-3), BC loop, CC' loop, or G chain of human TIM-3; (b) one, two, three or all of: A-chain, EF loop, F-chain or FG loop; (c) one, two, three, or all of: the third residue from the N-terminus of the A chain (Glu23 in human TIM-3), C chain, C'C'' loop, or C'' chain; (d) one or both of Val24 and Glu25 residues adjacent to the N-terminus of the A chain; the Thr41 residue in the BC loop; four, five, six, seven, or all of the Glu121, Lys122, Phe123, Asn124, Leu125, Lys126, Leu127, and Val128 residues in the G chain; and three, four, five, or all of the Gly56, Ala57, Cys58, Pro59, Val60, and Phe61 residues in the CC' loop; (e) Val24 and Glu25 residues adjacent to the N-terminus of the A chain; the Thr41 residue in the BC loop; residues Glu121, Lys122, Phe123, Asn124, Leu125, Lys126, Leu127 and Val128 in the G chain and residues Gly56, Ala57, Cys58, Pro59, Val60 and Phe61 in the CC' loop; (f) one or more residues selected from: a Tyr26 residue in the A chain, Phe39 and Tyr40 residues in the BC loop; Ser105 residue in the EF loop; residues Gly106 and Ile107 in the F chain; and Asn119 and Asp120 residues in the FG loop; (g) Tyr26 residue in the A chain, Phe39 and Tyr40 residues in the BC loop; Ser105 residues in the EF loop; Gly106 and Ile107 residues in the F chain and Asn119 and Asp120 residues in the FG loop; (h) one or more residues selected from: a Glu23 residue which is the N-terminal residue of the A chain; residues Pro42, Ala43, Ala44, Pro45, Gly46, Asn47, Leu48, Val49 and Pro50 in loop BC; Val51, Cys52, Trp53, Gly54 and Lys55 residues in the C chain; Arg73 and Asp74 residues in the C'C' loop and Val75, Asn76 and Tyr77 residues in the C'' chain; and/or (i) a Glu23 residue which is the N-terminal residue of the A chain; residues Pro42, Ala43, Ala44, Pro45, Gly46, Asn47, Leu48, Val49 and Pro50 in the BC loop; Val51, Cys52, Trp53, Gly54, and Lys55 residues in loop C; Arg73 and Asp74 residues in the C'C'' chain; and Val75, Asn76 and Tyr77 residues in the C'' chain. 18. A pharmaceutical composition for stimulating an immune response in an individual, comprising an isolated antibody molecule according to any one of claims 1 to 17 and a pharmaceutically acceptable carrier, excipient or stabilizer. 19. A pharmaceutical composition for the treatment of cancer in an individual, comprising the isolated antibody molecule according to any one of claims 1 to 17 and a pharmaceutically acceptable carrier, excipient or stabilizer. 20. A pharmaceutical composition for the treatment of an infectious disease in an individual, comprising an isolated antibody molecule according to any one of claims 1 to 17 and a pharmaceutically acceptable carrier, excipient or stabilizer. 21. An isolated nucleic acid encoding the heavy and light chain variable region of an antibody molecule according to any one of claims 1-17. 22. Isolated nucleic acid encoding an antibody molecule containing CDR 1-3 heavy - 169 040365 light chain and light chain CDR 1-3 of the antibody molecule according to any one of claims 1-17. 23. Nucleic acid according to claim 22, containing: a nucleotide sequence encoding a VH, wherein said nucleotide sequence contains any of SEQ ID NOs: 11, 17, 29, 33, 37, 45, 49, 53, 61, 69, 73, 77, 81, 85, 93, 101, 115 or 120, or at least 85% identical to any of SEQ ID NOs: 11, 17, 29, 33, 37, 45, 49, 53, 61, 69, 73, 77, 81, 85, 93, 101, 115 or 120, and/or a nucleotide sequence encoding VL, where the specified nucleotide sequence contains any of SEQ ID NO: 15, 21, 41, 57, 65, 89, 97, 105, 118, 123, 125 or 127, or at least least 85% identical to any of SEQ ID NOs: 15, 21, 41, 57, 65, 89, 97, 105, 118, 123, 125, or 127. 24. Nucleic acid according to claim 22, containing: a nucleotide sequence encoding a heavy chain, wherein said nucleotide sequence comprises any of SEQ ID NOs: 19, 29, 35, 39, 47, 51, 55, 63, 71, 75, 79, 83, 87, 95, 103, 117, or 122, or at least 85% identical to any of SEQ ID NOs: 19, 29, 35, 39, 47, 51, 55, 63, 71, 75, 79, 83, 87, 95, 103, 117, or 122; and / or a nucleotide sequence encoding a light chain, where the specified nucleotide sequence contains any of SEQ ID NO: 23, 43, 59, 67, 91, 99, 107, 119, 124, 126, 128, or at least 85% identical to any of SEQ ID NOs: 23, 43, 59, 67, 91, 99, 107, 119, 124, 126, 128. 25. An expression vector containing a nucleic acid according to any one of claims 21-24. 26. A host cell containing a nucleic acid according to any one of claims 21-24, to obtain an antibody molecule according to any one of claims 1-17. 27. A method for producing an antibody molecule or fragment thereof, comprising culturing a host cell according to claim 26 under conditions suitable for gene expression. 28. A method for detecting TIM-3 in a biological sample, comprising (i) contacting the sample with an isolated antibody molecule according to any one of claims 1 to 17 under conditions that allow the interaction of the antibody molecule and the polypeptide, and (ii) detecting the formation of a complex between antibody molecule and sample. 29. The method according to claim 28, further comprising (iii) contacting a reference sample with an isolated antibody molecule according to any one of claims 1 to 17 under conditions that allow interaction of the antibody molecule and the polypeptide, and (ii) detecting the formation of a complex between the molecule antibodies and reference sample. 30. The use of an antibody molecule according to any one of claims 1 to 17 to stimulate an immune response in an individual. 31. The use of an antibody molecule according to any one of claims 1 to 17 for the treatment of a cancer in an individual. 32. The use of an antibody molecule according to any one of claims 1 to 17 for the treatment of an infectious disease in an individual. 33. Use according to claim 31, wherein the malignant tumor is lung cancer, melanoma, kidney cancer, liver cancer, prostate cancer, breast cancer, head and neck cancer, colon and rectal cancer, pancreatic cancer, brain cancer, hematological malignant tumor or metastatic focus of a malignant tumor. 34. The use of claim 33, wherein (a) the lung cancer is non-small cell lung cancer (NSCLC), lung adenocarcinoma, squamous cell lung carcinoma, or small cell lung cancer; (b) the melanoma is expanded melanoma, unresectable melanoma, metastatic melanoma, BRAF-mutated melanoma, NRAS-mutated melanoma, cutaneous melanoma, or intraocular melanoma; (c) the kidney cancer is renal cell carcinoma (RCC), metastatic renal cell carcinoma, clear cell renal cell carcinoma (CCRCC), clear cell renal cell carcinoma, or renal papillary cell carcinoma; or (d) the hematologic malignancy is lymphoma, non-Hodgkin's lymphoma, myeloma, or leukemia. 35. Use according to any one of claims 30-34, wherein the use further comprises the use of a second therapeutic agent, wherein the second therapeutic agent is one or more of chemotherapy, targeted cancer therapy, an oncolytic drug, a cytotoxic agent, an immune-based therapy, cytokine, surgery, radiotherapy, costimulatory molecule activator, inhibitor of inhibitory molecule, vaccine, or cellular immunotherapy. 36. Use according to any one of claims 30-35, characterized in that the use further comprises the use of: (a) a costimulatory molecule agonist selected from one or more 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 ligand; or - 170 040365 (b) an inhibitor of an immune control point molecule selected from one or more of PD-1, PD-L1, PD-L2, CTLA-4, LAG-3, CEACAM-1, CEACAM-5, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 or TGFR. 37. Use according to any one of claims 30-36, characterized in that the use further comprises the use of one or more of the following: (a) a PD-1 inhibitor; (b) a PD-L1 inhibitor; (c) a LAG-3 inhibitor; (d) a GITR agonist; (e) chemotherapy for the treatment of lung cancer; (f) an indolaminepyrrole-2,3-dioxygenase (IDO) inhibitor for the treatment of lung cancer; (g) a CTLA-4 inhibitor for the treatment of lung cancer or melanoma; (h) a MEK inhibitor for the treatment of lung cancer, melanoma or kidney cancer; (i) cancer vaccines; (j) one or more of: (i) immune therapy; (ii) directed funds; (iii) a VEGF tyrosine kinase inhibitor; (iv) an RNAi inhibitor; or (v) an inhibitor of the following mediator of VEGF signaling for the treatment of kidney cancer; (k) one, two or all of oxaliplatin, leucovorin or 5-FU for the treatment of lung cancer, melanoma or kidney cancer; or (l) a tyrzine kinase inhibitor for the treatment of kidney cancer. 38. Use according to claim 37, wherein one or more of the following: (a) a PD-1 inhibitor, which is an anti-PD-1 antibody molecule or is MDX-1106, Merck 3475, CT-011, AMP-224, or AMP-514; (b) a PD-L1 inhibitor which is an anti-PD-L1 antibody molecule or is YW243.55.S70, MPDL3280A, MEDI-4736, MSB-0010718C or MDX-1105; (c) a LAG-3 inhibitor, which is an anti-LAG-3 antibody molecule; (d) a GITR agonist, which is an anti-GITR antibody molecule; (e) chemotherapy, which is platinum doublet therapy; (f) an indolaminepyrrole-2,3-dioxygenase (IDO) inhibitor, which is INCB24360; (g) a CTLA-4 inhibitor, which is an anti-CTLA-4 antibody molecule, where the anti-CTLA-4 antibody molecule is ipilimumab, or a soluble CTLA-4 ligand, where the antibody molecule or pharmaceutical composition is used in combination with a BRAF inhibitor, where the inhibitor BRAF is vemurafenib or dabrafenib; (h) a MEK inhibitor for the treatment of lung cancer, melanoma or kidney cancer; (i) a cancer vaccine, which is a dendritic cell vaccine; (j) one or more of: (i) immune therapy, which is interleukin-2 or interferon-α; (ii) a targeting agent which is a VEGF inhibitor, wherein the VEGF inhibitor is an anti-VEGF antibody molecule; (iii) a VEGF tyrosine kinase inhibitor which is sunitinib, sorafenib, axitinib or pazopanib; (iv) an RNA-i inhibitor; or (v) an inhibitor of the following mediator of VEGF signaling, which is a mammalian target inhibitor of rapamycin (mTOR), wherein the mTOR inhibitor is everolimus or temsirulimus; (k) one, two or all of oxaliplatin, leucovorin or 5-FU for the treatment of lung cancer, melanoma or kidney cancer; or (l) a tyrzine kinase inhibitor, which is axitinib. 39. Use according to any one of claims 30-38, wherein the individual has or is identified as having one or more of: (a) a cancer that expresses TIM-3; (b) a cancer that is positive for one, two, or all of PD-L1, CD8, or IFN-γ; (c) a cancer that is triple positive for PD-L1, CD8 and IFN-γ; or (d) a cancer that is positive for tumor infiltrating lymphocytes (TIL). 40. Use according to any one of claims 30-39, where the antibody molecule is intended for use in up to - 171 040365 ze approximately 1 to 30 mg/kg or 1 to 5 mg/kg. 41. Use according to any one of claims 30-40, wherein the antibody molecule is intended to be used from once a week to once every 2, 3 or 4 weeks. 42. Use according to any one of claims 30-39, wherein the antibody molecule is to be administered at a dose of about 10 to 20 mg/kg biweekly. 43. Use according to any one of claims 30-42, wherein the antibody molecule is used in combination with one or more of the following: 1) a protein kinase C (PKC) inhibitor; 2) inhibitor of heat shock protein 90 (HSP90); 3) an inhibitor of phosphoinositide 3-kinase (PI3K) and/or a target of rapamycin (mTOR); 4) a cytochrome P450 inhibitor (eg, a CYP17 inhibitor or a 17-alpha-hydroxylase/C17-20-lyase inhibitor); 5) an iron chelating agent; b) an aromatase inhibitor; 7) a p53 inhibitor, eg an inhibitor of the p53/Mdm2 interaction; 8) apoptosis inducer; 9) an angiogenesis inhibitor; 10) aldosterone synthase inhibitor; 11) a smoothened receptor inhibitor (SMO); 12) a prolactin receptor (PRLR) inhibitor; 13) Wnt signal transduction inhibitor; 14) a CDK4/6 inhibitor; 15) an inhibitor of fibroblast growth factor receptor 2 (FGFR2)/fibroblast growth factor receptor 4 (FGFR4); 16) an inhibitor of macrophage colony stimulating factor (MCSF); 17) an inhibitor of one or more of c-KIT, histamine release, Flt3 (eg FLK2/STK1) or PKC; 18) an inhibitor of one or more of VEGFR-2 (eg, FLK-1/KDR), PDGFR-beta, c-KIT, or Raf kinase C; 19) a somatostatin agonist and/or a growth hormone releasing inhibitor; 20) an anaplastic lymphoma kinase (ALK) inhibitor; 21) an insulin-like growth factor receptor 1 (IGF-1R) inhibitor; 22) inhibitor of P-glycoprotein 1; 23) an inhibitor of vascular endothelial growth factor receptor (VEGFR); 24) BCR-ABL kinase inhibitor; 25) FGFR inhibitor; 26) CYP11B2 inhibitor; 27) an HDM2 inhibitor, eg an HDM2-p53 interaction inhibitor; 28) a tyrosine kinase inhibitor; 29) c-MET inhibitor; 30) a JAK inhibitor; 31) a DAC inhibitor; 32) 11b-hydroxylase inhibitor; 33) an IAP inhibitor; 34) a PIM kinase inhibitor; 35) a porcupine inhibitor; 36) a BRAF inhibitor, eg V600E BRAF or wild-type BRAF; 37) HER3 inhibitor; 38) MEK inhibitor; or 39) a lipid kinase inhibitor. 44. Use according to any one of claims 30-43, where the antibody molecule is used in combination with one or more of the compounds A1-A51, as described in table. 6. 45. Use according to any one of claims 30-44, wherein the antibody molecule is used in combination with azacitidine and/or decitabine. 46. The use of claim 45 wherein the antibody molecule is used to treat acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS). 47. An antibody molecule capable of binding to human TIM-3, comprising a VH comprising the amino acid sequence of SEQ ID NO: 52 and a VL comprising the amino acid sequence of SEQ ID NO: 64. 48. An antibody molecule capable of binding to human TIM-3, comprising a VH comprising the amino acid sequence of SEQ ID NO: 32 and a VL comprising the amino acid sequence of SEQ ID NO: 20. 49. An antibody molecule capable of binding to human TIM-3, comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 54 and a light chain comprising the amino acid sequence of SEQ ID NO: 66. 50. An antibody molecule capable of binding to human TIM-3, comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 34 and a light chain comprising the amino acid sequence of SEQ ID NO: 22. 51. Use of an antibody molecule capable of binding to TIM-3, comprising a VH containing the amino acid sequence of SEQ ID NO: 52 and a VL containing the amino acid sequence of SEQ ID NO: 64, for the treatment of cancer in an individual. 52. Use of an antibody molecule capable of binding to TIM-3, comprising a VH containing the amino acid sequence of SEQ ID NO: 32 and a VL containing the amino acid sequence of SEQ ID NO: 20, for the treatment of cancer in an individual. 53. Use of an antibody molecule capable of binding to TIM-3, comprising a VH containing the amino acid sequence of SEQ ID NO: 54 and a VL containing the amino acid sequence of SEQ ID NO: 66, for the treatment of cancer in an individual. 54. Use of an antibody molecule capable of binding to TIM-3, comprising a VH containing the amino acid sequence of SEQ ID NO: 34 and a VL containing the amino acid sequence of SEQ ID NO: 22, for the treatment of cancer in an individual. - 172 040365 Mouse antibody variable region sequence ABTIM3 Heavy chain (118 aa; SEQ ID NO: 1) QVQLQQPGAE LVKPGASVKM SCKASGYTFT |SYNMH|WIKQT PGQGLEWIGp IYPGNGDTSY| |NQKFKG|KATL TADKSSSTVY MQLSSLTSED SAVYYCAR|VG GAFPMDT^GQ GTSVTVSS Light chain (118 aa; SEQ ID NO: 2) DIVLTQSPAS LAVSLGQRAT ISC^ASESVE YYGTSLMQbY QQKPGQPPKL LIY|AASNVE~s| GVPARFSGSG SGTDFSLNIH PVEEDDIAIY FS|QQSRKDPS~T|FGGGTKLEI K Fig. 1A Sequence Alignment of ABTIM3 Mouse Antibody Against Mouse Germline Antibody Sequences