Affinity matured anti-OX40 antibodies and use thereof
Provided herein are high affinity antagonistic anti-OX40 antibodies, methods of making such antibodies, and methods of using such antibodies to treat OX40-mediated disorders.
1 . An antibody, or antigen binding fragment thereof, that binds to OX40 comprising:
a) a heavy chain variable region comprising a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:1, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:2, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:3; and
b) a light chain variable region comprising a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:4, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:5, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:38 or 71, where X 1 is F, T, W, or M; where X 2 is G, I, V, L, or E; where X 3 is A, D, E, L, H, T, or F; and where X 4 is W, P, F, Y, or T.
2 . The antibody, or antigen binding fragment thereof, of claim 1 , wherein the CDR-L3 comprises the amino acid sequence set forth in SEQ ID NO:38.
3 . The antibody, or antigen binding fragment thereof, of claim 1 , wherein X 1 is F, X 2 is G, X 3 is A, and X 4 is W or CDR-L3 comprises the amino acid sequence of SEQ ID NO:6.
4 . The antibody, or antigen binding fragment thereof, of claim 1 , wherein CDR-L3 comprises the amino acid sequence of SEQ ID NO:11, 14, 17, 20, 23, 26, or 29.
5 . The antibody or antigen binding fragment thereof that binds to OX40 of claim 1 , wherein the antibody is selected from:
a) an antibody comprising three heavy chain complementarity determining regions (CDRs) (CDR-H1, CDR-H2, and, CDR-H3) of a heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO:7 and three light chain complementarity determining regions (CDRs) (CDR-L1, CDR-L2, and CDR-L3), of a light chain variable domain (VL) comprising the amino acid sequence of SEQ ID NO:8;
b) an antibody comprising three heavy chain complementarity determining regions (CDRs) (CDR-H1, CDR-H2, and CDR-H3) of a heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO:7 and three light chain complementarity determining regions (CDRs) (CDR-L1, CDR-L2, and CDR-L3), of a light chain variable domain (VL) comprising the amino acid sequence of SEQ ID NO:12;
c) an antibody comprising three heavy chain complementarity determining regions (CDRs) (CDR-H1, CDR-H2, and CDR-H3) of a heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO:7 and three light chain complementarity determining regions (CDRs) (CDR-L1, CDR-L2, and CDR-L3), of a light chain variable domain (VL) comprising the amino acid sequence of SEQ ID NO:15
d) an antibody comprising three heavy chain complementarity determining regions (DR s) (CDR-H1, CDR-H2, and CDR-H3) of a heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO:7 and three light chain complementarity determining regions (CDRs) (CDR-L1, CDR-L2, and CDR-L3), of a light chain variable domain (VL) comprising the amino acid sequence of SEQ ID NO:18;
e) an antibody comprising three heavy chain complementarity determining regions (DR s) (CDR-H1, CDR-H2, and CDR-H3) of a heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO:7 and three light chain complementarity determining regions (CDRs) (CDR-L1, CDR-L2, and CDR-L3), of a light chain variable domain (VL) comprising the amino acid sequence of SEQ ID NO:21;
f) an antibody comprising three heavy chain complementarity determining regions (CDRs) (CDR-H1, CDR-H2, and CDR-H3) of a heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO:7 and three light chain complementarity determining regions (CDRs) (CDR-L1, CDR-L2, and CDR-L3), of a light chain variable domain (VL) comprising the amino acid sequence of SEQ ID NO:24;
g) an antibody comprising three heavy chain complementarity determining regions (CDRs) (CDR-H1, CDR-H2, and CDR-H3) of a heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO:7 and three light chain complementarity determining regions (CDRs) (CDR-L1, CDR-L2, and CDR-L3), of a light chain variable domain (VL) comprising the amino acid sequence of SEQ ID NO:27; or
h) an antibody comprising three heavy chain complementarity determining regions (CDRs) (CDR-H1, CDR-H2, and CDR-H3) of a heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO:7 and three light chain complementarity determining regions (CDRs) (CDR-L1, CDR-L2, and CDR-L3), of a light chain variable domain (VL) comprising the amino acid sequence of SEQ ID NO:30.
6 . The antibody or antigen binding fragment thereof that binds to OX40 of claim 5 , comprising three heavy chain complementarity determining regions (CDRs) (CDR-H1, CDR-H2, and CDR-H3) of a heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO:7 and three light chain complementarity determining regions (CDRs) (CDR-L1, CDR-L2, and CDR-L3), of a light chain variable domain (VL) comprising the amino acid sequence of SEQ ID NO:8.
7 . The antibody or antigen binding fragment of claim 5 , wherein:
a) the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to Kabat;
b) the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to Chothia;
c) the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to IMGT;
d) the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to AbM;
e) the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to Contact; or
f) the CDR-H1, CDR-H2, and CDR-H3, and the CDR-L1, CDR-L2, and CDR-L3 are defined according to Honneger (AHo).
8 . The antibody or antigen binding fragment thereof of claim 5 , wherein:
the heavy chain CDR-H1 comprises the amino acid sequence of SEQ ID NO:1, the heavy chain CDR-H2 comprises the amino acid sequence of SEQ ID NO:2, the heavy chain CDR-H3 comprises the amino acid sequence of SEQ ID NO:3, the light chain CDR-L1 comprises the amino acid sequence of SEQ ID NO:4, the light chain CDR-L2 comprises the amino acid sequence of SEQ ID NO:5, and the light chain CDR-L3 comprise the amino acid sequence of SEQ ID NO:6.
9 . The antibody or antigen binding fragment thereof of claim 5 , wherein the antibody or antibody or antigen binding fragment thereof comprises:
(a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:7 and/or a light chain variable region comprising the amino acid sequence of SEQ ID NO:8; or
(b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:7 and a light chain variable region comprising an amino acid sequence selected from SEQ ID NO:12, 15, 18, 21, 24, 27, or 30.
10 . The antibody or antigen binding fragment thereof of claim 5 , wherein the antibody or antigen binding fragment thereof comprises:
(a) a heavy chain Fc region comprising an amino acid modification that increases the serum half-life of the antibody or antigen binding fragment thereof;
(b) an IgG1 heavy chain; or
(c) an IgG1 heavy chain Fc region comprising the mutations M252Y, S254T, and T256E as numbered according to EU numbering.
11 . The antibody or antigen binding fragment thereof of claim 5 , wherein the antibody or antigen binding fragment thereof comprises:
(a) a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain comprising the amino acid sequence of SEQ ID NO:10, 13, 16, 19, 22, 25, 28, or 31;
(b) a heavy chain comprising the amino acid sequence of SEQ ID NO:73 and a light chain comprising the amino acid sequence of SEQ ID NO:13, 16, 19, 22, 25, 28, or 31; or
(c) a heavy chain comprising the amino acid sequence of SEQ ID NO:73 and a light chain comprising the amino acid sequence of SEQ ID NO:10.
12 . An isolated nucleic acid comprising a nucleotide sequence encoding the heavy chain complementarity determining regions and the light chain complementarity determining regions of the antibody or antigen binding fragment thereof of claim 5 .
13 . An expression vector comprising the nucleic acid of claim 12 .
14 . A host cell comprising the expression vector of claim 13 .
15 . A method of producing an antibody, or antigen binding fragment thereof, that binds to human OX40, the method comprising:
(a) growing the host cell of claim 14 under conditions so that the host cell expresses a polypeptide or polypeptides comprising the heavy chain or heavy chain variable region and a polypeptide or polypeptides comprising the light chain or light chain variable region, thereby producing the antibody or the antigen-binding fragment of the antibody; and
(b) purifying the antibody or the antigen-binding fragment of the antibody.
16 . A pharmaceutical composition comprising the antibody or antigen binding fragment of claim 5 , and a pharmaceutically acceptable carrier.
17 . The antibody or antigen binding fragment thereof of claim 5 , wherein the antibody or antigen binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:7 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:8.
18 . The antibody or antigen binding fragment thereof of claim 5 , wherein the antibody or antigen binding fragment thereof comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:73 and a light chain comprising the amino acid sequence of SEQ ID NO:10.