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 one 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.
1. 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: 6 or an amino acid sequence with at least 90% homology to SEQ ID NO: 6, an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 10 or an amino acid sequence with at least 90% homology to SEQ ID NO: 10, and an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 14 or an amino acid sequence with at least 90% homology to SEQ ID NO: 14, and the heavy chain variable domain comprises an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 19 or an amino acid sequence with at least 90% homology to SEQ ID NO: 19, an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22 or an amino acid sequence with at least 90% homology to SEQ ID NO: 22, and an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 26 or an amino acid sequence with at least 90% homology to SEQ ID NO: 26.
2. The anti-Siglec-9 antibody of claim 1 , wherein the anti-Siglec-9 antibody comprises
an HVR-L1 comprising the amino acid sequence of SEQ ID NO: 6, an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 10, an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 14, an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 19, an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 22, and an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 26.
3. The anti-Siglec-9 antibody of claim 2 , wherein the antibody is of the IgG class the IgM class, or the IgA class.
4. The anti-Siglec-9 antibody of claim 3 , wherein the anti-Siglec-9 antibody has an IgG1, IgG2, IgG3, or IgG4 isotype.
5. The anti-Siglec-9 antibody of claim 4 , 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.
6. The anti-Siglec-9 antibody of claim 2 , wherein the anti-Siglec-9 antibody is an antibody fragment that binds to human Siglec-9 or mammalian Siglec-9 protein.
7. The anti-Siglec-9 antibody of claim 6 , wherein the fragment is an Fab, Fab′, Fab′-SH, F(ab′)2, Fv, or scFv fragment.
8. The anti-Siglec-9 antibody of claim 2 , wherein the anti-Siglec-9 antibody is a murine antibody, a humanized antibody, a bispecific antibody, a monoclonal antibody, a multivalent antibody, a conjugated antibody, or a chimeric antibody.
9. The anti-Siglec-9 antibody of claim 2 , wherein the anti-Siglec-9 antibody has dissociation constant (KD) for human Siglec-9 and mammalian Siglec-9 that ranges from about 10 nM to about 10 pM, wherein the KD is determined at a temperature of approximately 25° C.
10. The anti-Siglec-9 antibody of claim 2 , wherein the anti-Siglec-9 antibody has dissociation constant (KD) for human Siglec-9 that ranges from about 9 nM to about 230 pM, wherein the KD is determined at a temperature of approximately 25° C.
11. The isolated anti-Siglec-9 antibody of claim 1 , wherein the anti-Siglec-9 antibody decreases cell surface levels of Siglec-9 without inhibiting interaction between Siglec-9 and one or more Siglec-9 ligands.
12. The anti-Siglec-9 antibody of claim 11 , wherein the antibody decreases cell surface levels of Siglec-9 in vivo.
13. The anti-Siglec-9 antibody of claim 11 , wherein the anti-Siglec-9 antibody inhibits one or more Siglec-9 activities.
14. The anti-Siglec-9 antibody of claim 11 , wherein the one or more Siglec-9 ligands are selected from the group consisting of Siglec-9 ligands expressed on red blood cells, Siglec-9 ligands expressed on bacterial cells, Siglec-9 ligands expressed on apoptotic cells, Siglec-9 ligands expressed on nerve cells, Siglec-9 ligands expressed on glia cells, Siglec-9 ligands expressed on microglia, Siglec-9 ligands expressed on astrocytes, Siglec-9 ligands expressed on tumor cells, Siglec-9 ligands expressed on viruses, Siglec-9 ligands expressed on dendritic cells, Siglec-9 ligands bound to beta amyloid plaques, Siglec-9 ligands bound to Tau tangles, Siglec-9 ligands on disease-causing proteins, Siglec-9 ligands on disease-causing peptides, Siglec-9 ligands expressed on macrophages, Siglec-9 ligands expressed on neutrophils, Siglec-9 ligands expressed on natural killer cells, Siglec-9 ligands expressed on monocytes, Siglec-9 ligands expressed on T cells, Siglec-9 ligands expressed on T helper cells, Siglec-9 ligands expressed on cytotoxic T cells, Siglec-9 ligands expressed on B cells, Siglec-9 ligands expressed on tumor-imbedded immunosuppressor dendritic cells, Siglec-9 ligands expressed on tumor-imbedded immunosuppressor macrophages, Siglec-9 ligands expressed on myeloid-derived suppressor cells, Siglec-9 ligands expressed on regulatory T cells, secreted mucins, sialic acid, sialic acid-containing glycolipids, sialic acid-containing glycoproteins, alpha-2,8-disialyl containing glycolipids, branched alpha-2,6-linked sialic acid-containing glycoproteins, terminal alpha-2,6-linked sialic acid-containing glycolipids, terminal alpha-2,3-linked sialic acid-containing glycoproteins, and di sialogangliosides.
15. The anti-Siglec-9 antibody of claim 11 , wherein the antibody decreases cell surface levels of Siglec-9 in vitro on primary cells selected from the group consisting of dendritic cells, bone marrow-derived dendritic cells, monocytes, microglia, macrophages, neutrophils, and NK cells, or on cell lines.
16. The anti-Siglec-9 antibody of claim 11 , wherein the anti-Siglec-9 antibody does not reduce TREM2 expression.
17. The anti-Siglec-9 antibody of claim 11 , wherein the anti-Siglec-9 antibody binds to one or more amino acids within
amino acid residues 185-194 of SEQ ID NO: 1, or amino acid residues on a mammalian Siglec-9 protein corresponding to amino acid residues 185-194 of SEQ ID NO: 1.
18. The anti-Siglec-9 antibody of claim 11 , wherein the anti-Siglec-9 antibody binds to one or more amino acid residues selected from the group consisting of D189, P190, and R194 of SEQ ID NO: 1, or one or more amino acid residues on a mammalian Siglec-9 protein corresponding to an amino acid residue selected from the group consisting of D189, P190, and R194 of SEQ ID NO: 1.
19. The anti-Siglec-9 antibody of claim 11 , wherein the Siglec-9 protein is a mammalian protein or a human protein.
20. The anti-Siglec-9 antibody of claim 11 , wherein the anti-Siglec-9 antibody binds Siglec-9 in a pH dependent manner.
21. A pharmaceutical composition comprising the anti-Siglec-9 antibody of claim 2 , and a pharmaceutically acceptable carrier.
22. A humanized form of the anti-Siglec-9 antibody of claim 1 .
23. The anti-Siglec-9 antibody of claim 1 , wherein the heavy chain variable domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 116-126, and the light chain variable domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 61-71.