ANTI-TIGIT ANTIBODIES AND METHODS OF USE THEREOF
The instant disclosure provides antibodies that specifically bind to T-cell immunoreceptor with Ig and ITIM domains (TIGIT) (e.g., human TIGIT) and antagonize TIGIT function. Also provided are pharmaceutical compositions comprising these antibodies, nucleic acids encoding these antibodies, expression vectors and host cells for making these antibodies, and methods of treating a subject using these antibodies.
1 . An isolated antibody that specifically binds to human TIGIT, the antibody comprising a heavy chain variable region comprising complementarity determining regions (CDRs) CDRH1, CDRH2, and CDRH3, and a light chain variable region comprising complementarity determining regions CDRL1, CDRL2, and CDRL3, wherein:
(a) CDRH1 comprises the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2;
(b) CDRH2 comprises the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 4;
(c) CDRH3 comprises the amino acid sequence of SEQ ID NO: 5;
(d) CDRL1 comprises the amino acid sequence of SEQ ID NO: 6;
(e) CDRL2 comprises the amino acid sequence of SEQ ID NO: 7; and/or
(f) CDRL3 comprises the amino acid sequence of SEQ ID NO: 8.
2 . The isolated antibody of claim 1 , wherein CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 comprise the amino acid sequences set forth in SEQ ID NOs: 1, 3, 5, 6, 7, and 8; or 2, 4, 5, 6, 7, and 8, respectively.
3 . (canceled)
4 . The isolated antibody of claim 1 , wherein:
(a) the heavy chain variable region comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 9: optionally wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 9, optionally wherein X in SEQ ID NO: 9 is glutamate (E) or pyroglutamate (pE); and/or
(b) the light chain variable region comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 10; optionally wherein the light chain variable region comprises the amino acid of SEQ ID NO: 10, optionally wherein X in SEQ ID NO: 9 is glutamate (E) or pyroglutamate (pE).
5 - 22 . (canceled)
23 . The isolated antibody of claim 1 , wherein:
(a) the heavy chain variable region comprises an amino acid sequence derived from a human IGHV1-69*01 germline sequence,
(b) the heavy chain variable region comprises an amino acid sequence derived from a human IGHV1-69*06 germline sequence:
(c) the heavy chain variable region comprises an amino acid sequence derived from a human IGHV1-69*12 germline sequence:
(d) the light chain variable region comprises an amino acid sequence derived from a human IGLV2-14*01 germline sequence:
(e) the light chain variable region comprises an amino acid sequence derived from a human IGLV2-23*02 germline sequence; and/or
(f) the light chain variable region comprises an amino acid sequence derived from a human IGLV2-11*01 germline sequence.
24 - 28 . (canceled)
29 . The isolated antibody of claim 1 wherein:
(a) the heavy chain variable region comprises an amino acid region that is at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 34 or 35, optionally wherein the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 9; and/or
(b) the light chain variable region comprises an amino acid region that is at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOs: 37-39 and 60, optionally wherein the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 10.
30 - 31 . (canceled)
32 . An isolated antibody that specifically binds to human TIGIT, wherein:
(a) the antibody binds to the same epitope of human TIGIT as an antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 9 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 10,
(b) the antibody binds to an epitope located within a region of human TIGIT, the amino acid sequence of the region consisting of the amino acid sequence of any one of SEQ ID NOs: 31-33:
(c) the antibody binds to one or more amino acid residues of human TIGIT selected from the group consisting of Q35, 147, N49, H90, and T96, numbered according to the amino acid sequence of SEQ ID NO: 40:
(d) the antibody does not bind to one or more of the amino acid residues selected from the group consisting of T34, L52, H55, 156, S57, P58, S59, T98, R100, and F102 of human TIGIT, numbered according to the amino acid sequence of SEQ ID NO: 40;
(e) the binding of the antibody to a protein comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 36, 44, 45, 46, 48, 51, 52, 53, 57, and 59 is substantially weakened relative to the binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 42; and/or
(f) the binding of the antibody to a protein comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 43, 47, 49, 54, 55, 56, and 58 is not substantially weakened relative to the binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 42.
33 - 106 . (canceled)
107 . The isolated antibody of claim 1 ,
wherein the antibody further comprises a heavy chain constant region selected from the group consisting of human IgG 1 , IgG 2 , IgG 3 , IgG 4 , IgA 1 , and IgA 2 , optionally wherein:
(a) the antibody comprises an IgG 1 heavy chain constant region, optionally wherein
(1) the IgG 1 heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 19,
(2) the amino acid sequence of the IgG 1 heavy chain constant region comprises an N297A mutation, numbered according to the EU numbering system,
(3) the IgG 1 heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 20,
(4) the amino acid sequence of the IgG 1 heavy chain constant region comprises L234F, L235F, and N297A mutations, numbered according to the EU numbering system, (5) the IgG 1 heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 21,
(6) the amino acid sequence of the IgG 1 heavy chain constant region comprises S239D and 1332E mutations, numbered according to the EU numbering system,
(7) the IgG 1 heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 22,
(8) the amino acid sequence of the IgG 1 heavy chain constant region comprises S239D, A330L, and 1332E mutations, numbered according to the EU numbering system,
(9) the IgG 1 heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 23,
(10) the amino acid sequence of the IgG 1 heavy chain constant region comprises L235V, F243L, R292P, Y300L, and P396L mutations, numbered according to the EU numbering system,
(11) the IgG 1 heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 24,
(12) the amino acid sequence of the IgG 1 heavy chain constant region comprises S267E and L328F mutations, numbered according to the EU numbering system,
(13) the IgG 1 heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 25, and/or
(14) the IgG 1 heavy chain constant region is afucosylated;
(b) the antibody comprises an IgG 4 heavy chain constant region, optionally wherein
(1) the amino acid sequence of the IgG 4 heavy chain constant region comprises a S228P mutation, numbered according to the EU numbering system, and/or
(2) the IgG 4 heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 26; and/or
(c) antibody comprises a light chain constant region comprising the amino acid sequence of SEQ ID NO: 28.
108 - 122 . (canceled)
123 . The isolated antibody of claim 107 , wherein the increase of FcγRIIIA and/or FcγRIIA activity in a first cytotoxic cell contacted with the antibody is greater than the increase of FcγRIIIA and/or FcγRIIA activity in a second cytotoxic cell contacted with a reference antibody comprising the same heavy chain variable region as the antibody, and a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 19, wherein the cytotoxic cell is optionally a natural killer cell.
124 - 128 . (canceled)
129 . The isolated antibody of claim 1 comprising:
(a) a heavy chain comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 11-18; and/or
(b) a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27.
130 . (canceled)
131 . The isolated antibody of claim 1 , wherein:
(a) the antibody is a human antibody;
(b) the antibody is a bispecific antibody:
(c) the antibody is antagonistic to human TIGIT;
(d) the antibody preferentially kills regulatory T cells over effector T cells in a population of peripheral blood mononuclear cells (PBMCs) in vitro:
(e) the antibody decreases or inhibits binding of human TIGIT to PVR or PVRL2 relative to the level of binding in the absence of the antibody:
(f) the antibody induces IL-2 and/or IFNγ production by PBMCs stimulated with staphylococcal enterotoxin A (SEA):
(g) the antibody is conjugated to a cytotoxic agent, cytostatic agent, toxin, radionuclide, or detectable label:
(h) the antibody is cross-linked to a second antibody or a fragment thereof; and/or
(i) the antigen-binding fragment specifically binds to human TIGIT.
132 - 139 . (canceled)
140 . A pharmaceutical composition comprising the antibody of claim 1 and a pharmaceutically acceptable carrier or excipient.
141 . An isolated polynucleotide encoding a heavy chain and/or light chain of the antibody of claim 1 .
142 . A vector comprising the polynucleotide of claim 141 .
143 . A recombinant host cell comprising the polynucleotide of claim 141 .
144 . A method of producing an antibody that specifically binds to human TIGIT, or an antigen-binding fragment thereof, the method comprising culturing the host cell of claim 143 such that the polynucleotide is expressed and the antibody, or antigen-binding fragment, is produced.
145 . A method of:
(a) increasing T cell activation in response to an antigen in a subject;
(b) decreasing or inhibiting Treg activity in response to an antigen in a subject; and/or
(c) increasing NK cell activation in response to an antigen in a subject,
the method comprising administering to the subject an effective amount of the antibody of claim 1 .
146 - 147 . (canceled)
148 . A method of:
(a) treating cancer in a subject; or
(b) treating an infectious disease in a subject,
the method comprising administering to the subject an effective amount of the antibody of claim 1 , optionally wherein the antibody is administered intravenously, subcutaneously, intratumorally, or is delivered to a tumor draining lymph node.
149 - 168 . (canceled)
169 . A method of:
(a) increasing T cell activation in response to an antigen in a subject;
(b) decreasing or inhibiting Treg activity in response to an antigen in a subject; and/or
(c) increasing NK cell activation in response to an antigen in a subject, the method comprising administering to the subject an effective amount of the antibody of claim 32 .
170 . A method of:
(a) treating cancer in a subject; or
(b) treating an infectious disease in a subject,
the method comprising administering to the subject an effective amount of the antibody of claim 32 , optionally wherein the antibody is administered intravenously, subcutaneously, intratumorally, or is delivered to a tumor draining lymph node.