Anti-CD40 binding molecules and bi-specific antibodies comprising such
Provided herein are antibodies (e.g., humanized antibodies) binding to CD40 and bi-specific antibodies comprising such for targeting both CD40 and a second suitable antigen such as a tumor antigen or an immune checkpoint molecule. Examples of the second antigen include PD-1, PD-L1, HER2, B7H3, B7H4, netrotic tumor cells (TNT), or CEA. Also provided herein are therapeutic uses of such antibodies.
1 . A humanized antibody specific to human CD40, wherein the humanized antibody comprises a heavy chain variable region (V H ) and a light chain variable region (V L ), wherein:
(i) the V H comprises a framework of IGHV3-73*01 and heavy chain complementary determining regions (CDRs) 1, 2, and 3, and the V L comprises a framework of IGKV1-39*01 and light chain CDRs 1, 2, and 3; wherein
the V H comprises the heavy chain CDR1 comprising the amino acid sequence of GFNFNDSFMN (SEQ ID NO:1), GENFQDSFMN (SEQ ID NO:2), GENFNDAFMN (SEQ ID NO: 3), or GENENDYFMN (SEQ ID NO:4), the heavy chain CDR2 comprising the amino acid sequence of QIRNKNYNYATYYTESLEG (SEQ ID NO:5), and the heavy chain CDR3 comprising the amino acid sequence of YYYDGFADYFDY (SEQ ID NO:6); and the V L comprises the light chain CDR1, light chain CDR2, and light chain CDRs comprising the amino acid sequences of KASQNIYIYLN (SEQ ID NO:7), NTNNLOT (SEQ ID NO:8), and LOHSSRRT (SEQ ID NO:9), respectively; or
(ii) the V H comprises a framework of IGHV3-23*04 and heavy chain CDRs 1, 2, and 3, and the V L comprises a framework of IGKV1-39*01 and light chain CDRs 1, 2, and 3; wherein:
the V H comprises the heavy chain CDR1, the heavy chain CDR2, and the heavy chain CDR3 comprising the amino acid sequences of GFTFTNYGLH (SEQ ID NO:16), SISPSGGVTYYRDSVKG (SEQ ID NO:17), and PFLGWGGANWIAH (SEQ ID NO:18) respectively; and the V L comprises the light chain CDR1, the light chain CDR2, and the light chain CDR3 comprising the amino acid sequences of LASEDISNDLA (SEQ ID NO:19), FVDRLLD (SEQ ID NO:20), and QQSYKYPPT (SEQ ID NO:21), respectively.
2 . The humanized antibody of claim 1 , wherein the humanized antibody is set forth in (i).
3 . The humanized antibody of claim 2 , wherein the V H comprises one or more mutations in the V H framework.
4 . The humanized antibody of claim 3 , wherein the mutations in the V H framework are back mutations based on amino acid residues in the parent murine antibody at corresponding positions.
5 . The humanized antibody of claim 4 , wherein the back mutations comprise E1Q, A24T, F70V, R100S, or a combination thereof.
6 . The humanized antibody of claim 2 , wherein the V H comprises the amino acid sequence of any one of SEQ ID NOs: 10-14 and the V L comprises the amino acid sequence of SEQ ID NO:15.
7 . The humanized antibody of claim 1 , wherein the antibody is set forth in (ii).
8 . The humanized antibody of claim 7 , wherein the V H comprises the amino acid sequence of SEQ ID NO:22; and wherein the V L comprises the amino acid sequence of SEQ ID NO:23.
9 . The humanized antibody of claim 1 , wherein the humanized antibody is a full-length antibody.
10 . The humanized antibody of claim 9 , wherein the full-length antibody is an IgG/kappa molecule.
11 . The humanized antibody of claim 10 , wherein the full-length antibody comprises a heavy chain that is an IgG1, IgG2, or IgG4 chain.
12 . The humanized antibody of claim 11 , wherein the heavy chain comprises a mutated Fc region, which exhibits altered binding affinity or selectivity to an Fc receptor as relative to the wild-type counterpart.
13 . The humanized antibody of claim 10 , wherein the humanized antibody is selected from the group consisting of TM550, TM553, LP3771, LP3772, LP3773, TM738, TM739, TM740, and Ly181; or wherein the antibody is selected from the group consisting of TM559, LP3781, LP3782, and LP3783.
14 . A bi-specific antibody, comprising:
(a) a first antibody moiety that binds human CD40, which is set forth in claim 1 ; and
(b) a second antibody moiety that binds an antigen selected from the group consisting of HER2, necrotic tumor cells (TNT), carcinoembryonic antigen (CEA), PD-1, PD-L1, B7H3, and B7H4.
15 . The bi-specific antibody of claim 14 , wherein either the first antibody moiety or the second antibody moiety is in a single-chain antibody (scFv) format; and wherein the other antibody moiety is in a full-length antibody format comprising a heavy chain and a light chain.
16 . The bi-specific antibody of claim 15 , wherein the first antibody moiety that binds human CD40 is a scFv; wherein the second antibody moiety comprises a first polypeptide comprising an antibody heavy chain and a second polypeptide comprising an antibody light chain, and wherein the scFv is fused to either the first polypeptide or the second polypeptide.
17 . The bi-specific antibody of claim 14 , which is selected from the group consisting of Ly301, Ly303, Ly338, Ly339, Ly340, Ly341, Ly342, Ly343, Ly344, Ly345, Ly349, and Ly350;
which is selected from the group consisting of Ly610, Ly611, Ly612, Ly613, Ly614, Ly615, Ly616, Ly617, Ly801, Ly802, Ly803, Ly804, Ly805, Ly806, Ly807, Ly808, Ly809, Ly810, Ly811, Ly812, Ly813, Ly814, Ly815, and Ly816;
which is selected from the group consisting of Ly474, Ly475, Ly476, Ly477, Ly478, Ly479, Ly480, Ly481, Ly482, Ly483, Ly484, Ly485, Ly486, Ly487, Ly488, Ly489, Ly490, Ly491, Ly492, Ly493, Ly494, Ly495, Ly496, and Ly497;
which is selected from the group consisting of Ly401, Ly402, Ly403, Ly404, Ly405, Ly406, Ly407, Ly408, Ly409, Ly410, Ly411, Ly412, Ly413, Ly414, Ly415, Ly416, Ly417, Ly418, Ly419, Ly420, Ly421, Ly422, Ly423, and Ly424;
which is selected from the group consisting of Ly462, Ly463, Ly464, Ly465, Ly466, Ly467, Ly468, Ly469, Ly470, Ly471, Ly472, and Ly473;
which is selected from the group consisting of Ly517, Ly518, Ly519, Ly520, Ly606, Ly607, Ly608, Ly609, Ly817, Ly818, Ly819, and Ly820; or
which is selected from the group consisting of Ly618, Ly619, Ly620, Ly621, Ly622, Ly623, Ly624, Ly625, Ly821, Ly822, Ly823, Ly824, Ly825, Ly826, Ly827, Ly828, Ly829, Ly830, Ly831, Ly832, Ly833, Ly834, Ly835, and Ly836.
18 . A nucleic acid or a nucleic acid set, which collectively encodes an antibody specific to human CD40, wherein the antibody is set forth in claim 1 .
19 . A host cell, comprising the nucleic acid or nucleic acid set of claim 18 .
20 . A method for producing a humanized antibody, comprising:
(i) culturing the host cell of claim 19 under conditions allowing for expression of the humanized antibody; and
(ii) harvesting the humanized antibody thus produced.
21 . A pharmaceutical composition comprising an antibody of claim 1 and a pharmaceutically acceptable carrier.
22 . A method for modulating immune responses, comprising administering an effective amount of the humanized antibody of claim 1 or the pharmaceutical composition thereof to a subject in need thereof.