ANTI-SIGLEC-9 ANTIBODIES AND METHODS OF USE THEREOF
The present disclosure is generally directed to compositions that include antibodies, e.g., monoclonal, chimeric, humanized antibodies, antibody fragments, etc., that specifically bind on or more epitopes within a Siglec-9 protein, e.g., human Siglec-9 or a mammalian Siglec-9, and use of such compositions in preventing, reducing risk, or treating an individual in need thereof.
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4 . The isolated anti-Siglec-9 antibody of claim 25 , wherein the anti-Siglec-9 antibody decreases cell surface levels of Siglec-9 and inhibits interaction between Siglec-9 and one or more Siglec-9 ligands.
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22 . The anti-Siglec-9 antibody of claim 25 , wherein the anti-Siglec-9 antibody comprises an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 173, an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 175, an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 177, an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 179, an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 181, and an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 183.
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25 . An isolated anti-Siglec-9 antibody, wherein the anti-Siglec-9 antibody comprises a light chain variable domain and a heavy chain variable domain, wherein the light chain variable domain comprises an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 173, or an amino acid sequence with at least 90% homology to SEQ ID NO: 173, an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 175, or an amino acid sequence with at least 90% homology to SEQ ID NO: 175, an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 177, or an amino acid sequence with at least 90% homology to SEQ ID NO: 177, and the heavy chain variable domain comprises an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 179, or an amino acid sequence with at least 90% homology to SEQ ID NO: 179, an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 181, or an amino acid sequence with at least 90% homology to SEQ ID NO: 181, and an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 183, or an amino acid sequence with at least 90% homology to SEQ ID NO: 183.
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28 . The anti-Siglec-9 antibody of claim 25 , wherein the anti-Siglec-9 antibody comprises a light chain variable domain and a heavy chain variable domain, wherein
the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 201; and the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 210.
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47 . The anti-Siglec-9 antibody of claim 44 , wherein:
(a) the anti-Siglec-9 antibody has a human or mouse IgG1 isotype and comprises one or more amino acid substitutions in the Fc region at a residue position selected from the group consisting of: N297A, D265A, D270A, L234A, L235A, G237A, P238D, L328E, E233D, G237D, H268D, P271G, A330R, C226S, C229S, E233P, L234V, L234F, L235E, P331S, S267E, L328F, A330L, M252Y, S254T, T256E, N297Q, P238S, P238A, A327Q, A327G, P329A, K322A, T394D, and any combination thereof, wherein the numbering of the residues is according to EU numbering, or comprises an amino acid deletion in the Fc region at a position corresponding to glycine 236;
(b) the anti-Siglec-9 antibody has an IgG1 isotype and comprises an IgG2 isotype heavy chain constant domain 1(CH1) and hinge region, optionally wherein the IgG2 isotype CH1 and hinge region comprises the amino acid sequence of ASTKGPSVFP LAPCSRSTSE STAALGCLVK DYFPEPVTVS WNSGALTSGVHTFPAVLQSS GLYSLSSVVT VPSSNFGTQT YTCNVDHKPS NTKVDKTVERKCCVECPPCP (SEQ ID NO: 171), and optionally wherein the antibody Fc region comprises a S267E amino acid substitution, a L328F amino acid substitution, or both, and/or a N297A or N297Q amino acid substitution, wherein the numbering of the residues is according to EU numbering;
(c) the anti-Siglec-9 antibody has an IgG2 isotype and comprises one or more amino acid substitutions in the Fc region at a residue position selected from the group consisting of: P238S, V234A, G237A, H268A, H268Q, V309L, A330S, P331S, C214S, C232S, C233S, S267E, L328F, M252Y, S254T, T256E, H268E, N297A, N297Q, A330L, and any combination thereof, wherein the numbering of the residues is according to EU numbering;
(d) the anti-Siglec-9 antibody has a human or mouse IgG4 isotype and comprises one or more amino acid substitutions in the Fc region at a residue position selected from the group consisting of: L235A, G237A, S228P, L236E, S267E, E318A, L328F, M252Y, S254T, T256E, E233P, F234V, L234A/F234A, S228P, S241P, L248E, T394D, N297A, N297Q, L235E, and any combination thereof, wherein the numbering of the residues is according to EU numbering; or
(e) the anti-Siglec-9 antibody has a hybrid IgG2/4 isotype, and optionally wherein the antibody comprises an amino acid sequence comprising amino acids 118 to 260 of human IgG2 and amino acids 261 to 447 of human IgG4, wherein the numbering of the residues is according to EU numbering.
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57 . The anti-Siglec-9 antibody of claim 25 , wherein the anti-Siglec-9 antibody binds Siglec-9 in a pH dependent manner.
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60 . The anti-Siglec-9 antibody of claim 25 , wherein the anti-Siglec-9 antibody is an antibody fragment that binds to a human Siglec-9 or a mammalian Siglec-9 protein.
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63 . The anti-Siglec-9 antibody of claim 60 , wherein the fragment is an Fab, Fab′, Fab′-SH, F(ab′)2, Fv, or scFv fragment.
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65 . The anti-Siglec-9 antibody of claim 25 , wherein the anti-Siglec-9 antibody is a humanized antibody, a bispecific antibody, a monoclonal antibody, a multivalent antibody, a conjugated antibody, or a chimeric antibody.
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72 . The anti-Siglec-9 antibody of claim 25 , wherein the anti-Siglec-9 antibody has dissociation constant (K D ) for human Siglec-9 and mammalian Siglec-9 that ranges from about 10 nM to about 10 pM, or less than 10 pM, wherein the K D is determined at a temperature of approximately 25° C.
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74 . An isolated nucleic acid comprising a nucleic acid sequence encoding the anti-Siglec-9 antibody of claim 25 .
75 . A vector comprising the nucleic acid of claim 74 .
76 . An isolated host cell comprising the vector of claim 75 .
77 . A method of producing an anti-Siglec-9 antibody, comprising culturing the host cell of claim 76 so that the anti-Siglec-9 antibody is produced.
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110 . A method of detecting Siglec-9 in a sample, comprising contacting the sample with an anti-Siglec-9 antibody, and detecting antibody-bound Siglec-9 in the sample, wherein the antibody comprises a light chain variable domain comprising an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 173, or an amino acid sequence with at least 90% homology to SEQ ID NO: 173, an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 175, or an amino acid sequence with at least 90% homology to SEQ ID NO: 175, an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 177, or an amino acid sequence with at least 90% homology to SEQ ID NO: 177, and a heavy chain variable domain comprising an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 179, or an amino acid sequence with at least 90% homology to SEQ ID NO: 179, an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 181, or an amino acid sequence with at least 90% homology to SEQ ID NO: 181, and an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 183, or an amino acid sequence with at least 90% homology to SEQ ID NO: 183.
111 . The method of claim 110 , wherein the antibody comprises:
(a) a light chain variable domain comprising a VL FR1 comprising the amino acid sequence of SEQ ID NO: 185, an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 173, a VL FR2 comprising the amino acid sequence of SEQ ID NO: 187, an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 175, a VL FR3 comprising the amino acid sequence of SEQ ID NO: 189, an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 177, and a VL FR4 comprising the amino acid sequence of SEQ ID NO: 44; and a heavy chain variable domain comprising a VH FR1 comprising the amino acid sequence of SEQ ID NO: 191, an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 179, a VH FR2 comprising the amino acid sequence of SEQ ID NO: 193, an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 181, a VH FR3 comprising the amino acid sequence of SEQ ID NO: 195, an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 183, and a VH FR4 comprising the amino acid sequence of SEQ ID NO: 196; or
(b) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 201 and a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 210.
112 . The method of claim 110 , wherein the antibody is a murine IgG1, IgG2A, or IgG2B antibody.
113 . The method of claim 110 , wherein the sample is a biopsy specimen.
114 . The method of claim 110 , wherein the sample is obtained from a subject suffering from cancer.
115 . The method of claim 110 , wherein the sample is a human sample.