T cell receptor-like antibody agents specific for EBV latent membrane protein 2A peptide presented by human HLA

Patent metadata

Field	Value
Publication number	US-10501559-B2
Application number	US-201615735133-A
Country	US
Kind code	B2
Filing date	Jun 9, 2016
Priority date	Jun 9, 2015
Publication date	Dec 10, 2019
Grant date	Dec 10, 2019

Abstract

Described herein are antibodies, fragments thereof and multi-specific binding agents that bind an Epstein-Barr virus (EBV) latent membrane protein 2 (LMP2) peptide presented by HLA class I molecules, in particular, HLA-A02. Also provided herein are methods of using the same or compositions thereof for the detection, prevention and/or therapeutic treatment of diseases characterized by expression of an EBV-LMP2 peptide presented by HLA-A02, in particular, Burkit's lymphoma, Hodgkin's lymphoma and nasopharyngeal carcinoma.

First claim

The invention claimed is:

1. A human antibody agent that binds an EBV-LMP2/HLA peptide complex comprising
(a) heavy chain CDR1 of SEQ ID NO:31, heavy chain CDR2 of SEQ ID NO:33 and heavy chain CDR3 of SEQ ID NO:35; and light chain CDR1 of SEQ ID NO:73, light chain CDR2 of SEQ ID NO:75 and light chain CDR3 of SEQ ID NO:77;
(b) heavy chain CDR1 of SEQ ID NO:37, heavy chain CDR2 of SEQ ID NO:39 and heavy chain CDR3 of SEQ ID NO:41; and light chain CDR1 of SEQ ID NO:79, light chain CDR2 of SEQ ID NO:81 and light chain CDR3 of SEQ ID NO:83;
(c) heavy chain CDR1 of SEQ ID NO:43, heavy chain CDR2 of SEQ ID NO:45 and heavy chain CDR3 of SEQ ID NO:47; and light chain CDR1 of SEQ ID NO:85, light chain CDR2 of SEQ ID NO:87 and light chain CDR3 of SEQ ID NO:89;
(d) heavy chain CDR1 of SEQ ID NO:49, heavy chain CDR2 of SEQ ID NO:51 and heavy chain CDR3 of SEQ ID NO:53; and light chain CDR1 of SEQ ID NO:91, light chain CDR2 of SEQ ID NO:93 and light chain CDR3 of SEQ ID NO:95;
(e) heavy chain CDR1 of SEQ ID NO:55, heavy chain CDR2 of SEQ ID NO:57 and heavy chain CDR3 of SEQ ID NO:59; and light chain CDR1 of SEQ ID NO:97, light chain CDR2 of SEQ ID NO:99 and light chain CDR3 of SEQ ID NO:101;
(f) heavy chain CDR1 of SEQ ID NO:61, heavy chain CDR2 of SEQ ID NO:63 and heavy chain CDR3 of SEQ ID NO:65; and light chain CDR1 of SEQ ID NO:103, light chain CDR2 of SEQ ID NO:105 and light chain CDR3 of SEQ ID NO:107; or
(g) heavy chain CDR1 of SEQ ID NO:67, heavy chain CDR2 of SEQ ID NO:69 and heavy chain CDR3 of SEQ ID NO:71; and light chain CDR1 of SEQ ID NO:109, light chain CDR2 of SEQ ID NO:111 and light chain CDR3 of SEQ ID NO:113.

2. The human antibody agent of claim 1 , wherein the human antibody agent has a K D of about 2.0 to about 170 nM.

3. The human antibody agent of claim 1 , wherein the EBV-LMP2 peptide has an amino acid sequence that is or comprises CLGGLLTMV (SEQ ID NO:1).

4. The human antibody agent of claim 3 , wherein the human antibody agent interacts directly with amino acid residues at one or more positions selected from the group consisting of position 1, 2, 3, 4, 5, 6, 7, 8, and combinations thereof, in the EBV-LMP2 peptide.

5. The human antibody agent of claim 1 , wherein the human antibody agent interacts directly with amino acid residues at one or more positions selected from the group consisting of position 1, position 5, position 8, and combinations thereof, in the EBV-LMP2 peptide.

6. The human antibody agent of claim 1 , wherein the human antibody agent comprises
(a) a heavy chain variable region sequence as set forth in any one of SEQ ID NOs: 5, 9, 13, 17, 21, 25, and 29, and
(b) a light chain variable region sequence as set forth in any one of SEQ ID NOs: 3, 7, 11, 15, 19, 23, and 27.

7. The human antibody agent of claim 1 , wherein the human antibody agent comprises
(a) the light chain variable region of SEQ ID NO:3 and the heavy chain variable region of SEQ ID NO:5;
(b) the light chain variable region of SEQ ID NO:7 and the heavy chain variable region of SEQ ID NO:9;
(c) the light chain variable region of SEQ ID NO:11 and the heavy chain variable region of SEQ ID NO:13;
(d) the light chain variable region of SEQ ID NO:15 and the heavy chain variable region of SEQ ID NO:17;
(e) the light chain variable region of SEQ ID NO:19 and the heavy chain variable region of SEQ ID NO:21;
(f) the light chain variable region of SEQ ID NO:23 and the heavy chain variable region of SEQ ID NO:25; or
(g) the light chain variable region of SEQ ID NO:27 and the heavy chain variable region of SEQ ID NO:29.

8. The human antibody agent of claim 1 , wherein the human antibody agent is a human monoclonal antibody that is defucosylated and/or glycosylated with terminal mannose, N-acetylglucose or glucose.

9. The human antibody agent of claim 1 , which human antibody agent comprises one or more amino acid substitutions that increase affinity to the EBV-LMP2/HLA peptide complex, wherein the one or more amino acid substitutions is selected from a light chain I48V substitution, a light chain K66R substitution, a heavy chain V5E substitution, a heavy chain E10D substitution, a heavy chain G26E substitution, and a heavy chain V78A substitution.

10. The human antibody agent of claim 9 , which human antibody agent that comprises one or more amino acid substitutions is characterized by
(a) a KA of at least about 1.6 higher affinity than affinity to a reference peptide/HLA complex, and/or
(b) no cross-reactivity to a reference peptide/HLA complex.

11. The human antibody agent of claim 1 , wherein the human antibody agent is a human monoclonal antibody or a fragment thereof.

12. The human antibody agent of claim 11 , wherein the human monoclonal antibody is an IgG1.

13. The human antibody agent of claim 11 , wherein the human monoclonal antibody fragment is an scFv.

14. A bispecific antibody comprising a first antigen-binding site that binds an EBV-LMP2/HLA peptide complex and a second antigen-binding site, wherein the first antigen-binding site comprises
(a) heavy chain CDR1 of SEQ ID NO:31, heavy chain CDR2 of SEQ ID NO:33 and heavy chain CDR3 of SEQ ID NO:35; and light chain CDR1 of SEQ ID NO:73, light chain CDR2 of SEQ ID NO:75 and light chain CDR3 of SEQ ID NO:77;
(b) heavy chain CDR1 of SEQ ID NO:37, heavy chain CDR2 of SEQ ID NO:39 and heavy chain CDR3 of SEQ ID NO:41; and light chain CDR1 of SEQ ID NO:79, light chain CDR2 of SEQ ID NO:81 and light chain CDR3 of SEQ ID NO:83;
(c) heavy chain CDR1 of SEQ ID NO:43, heavy chain CDR2 of SEQ ID NO:45 and heavy chain CDR3 of SEQ ID NO:47; and light chain CDR1 of SEQ ID NO:85, light chain CDR2 of SEQ ID NO:87 and light chain CDR3 of SEQ ID NO:89;
(d) heavy chain CDR1 of SEQ ID NO:49, heavy chain CDR2 of SEQ ID NO:51 and heavy chain CDR3 of SEQ ID NO:53; and light chain CDR1 of SEQ ID NO:91, light chain CDR2 of SEQ ID NO:93 and light chain CDR3 of SEQ ID NO:95;
(e) heavy chain CDR1 of SEQ ID NO:55, heavy chain CDR2 of SEQ ID NO:57 and heavy chain CDR3 of SEQ ID NO:59; and light chain CDR1 of SEQ ID NO:97, light chain CDR2 of SEQ ID NO:99 and light chain CDR3 of SEQ ID NO:101;
(f) heavy chain CDR1 of SEQ ID NO:61, heavy chain CDR2 of SEQ ID NO:63 and heavy chain CDR3 of SEQ ID NO:65; and light chain CDR1 of SEQ ID NO:103, light chain CDR2 of SEQ ID NO:105 and light chain CDR3 of SEQ ID NO:107; or
(g) heavy chain CDR1 of SEQ ID NO:67, heavy chain CDR2 of SEQ ID NO:69 and heavy chain CDR3 of SEQ ID NO:71, and light chain CDR1 of SEQ ID NO:109, light chain CDR2 of SEQ ID NO:111 and light chain CDR3 of SEQ ID NO:113.

15. The bispecific antibody of claim 14 , wherein the bispecific antibody is characterized by an EC 50 of about 0.2 to about 135 pM in T2 cells pulsed with EBV-LMP2 peptide CLGGLLTMV (SEQ ID NO:1).

16. The bispecific antibody of claim 14 , wherein the first and/or second antigen-binding sites are selected from the group consisting of an immunoglobulin molecule, scFv, scFab, Fab, Fv or a combination thereof.

17. The bispecific antibody of claim 14 , wherein the first and second antigen-binding sites are configured such that they form a single polypeptide chain.

18. The bispecific antibody of claim 17 , wherein the first and second antigen-binding sites are each scFvs.

19. The bispecific antibody of claim 14 , wherein the second antigen-binding site binds an immunological cell selected from the group consisting of a T cell, B cell, NK cell, dendritic cell, monocyte, macrophage, neutrophil, mesenchymal stem cell and neural stem cell.

20. The bispecific antibody

Assignees

• Memorial Sloan Kettering Cancer Center
• Eureka Therapeutics Inc

Inventors

• Cheung Nai-Kong V
• Ahmed Mahiuddin
• Lopez-Albaitero Andres
• Liu Cheng
• Yan Su
• Xiang Jingyi
• Liu Hong

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• C07K16/2809

  against the T-cell receptor (TcR)-CD3 complex · CPC title

• C07K2317/622

  Single chain antibody (scFv) · CPC title

• C07K2317/21

  from primates, e.g. man · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title