CN-115066518-B - Antibodies that bind to adenosine A2A receptor and methods of using the same for treating cancer and neurological diseases

Abstract

Provided herein are methods and compositions related to an adenosine A2A receptor library having nucleic acids encoding scaffolds comprising adenosine A2A binding domains. The adenosine A2A receptor libraries described herein encode immunoglobulins, including antibodies and single domain antibodies. Libraries described herein include diverse libraries comprising nucleic acids, each nucleic acid encoding a predetermined variant of at least one predetermined reference nucleic acid sequence. Further described herein are libraries of proteins produced when the nucleic acid library is translated. Further described herein are libraries of cells expressing the diverse nucleic acid libraries described herein.

Inventors

• AARON SATO
• Ray tabbiaza
• Pankaj gag
• LIU QIANG

Assignees

• 特韦斯特生物科学公司
• 特韦斯特生物科学公司

Dates

Publication Date

20260421

Application Date

20201209

Priority Date

20191209

Claims (13)

1. 1. An antibody or antibody fragment thereof that binds an adenosine A2A receptor, the antibody or antibody fragment thereof comprising an immunoglobulin heavy chain comprising a heavy chain variable region (VH) and an immunoglobulin light chain comprising a light chain variable region (VL), wherein: a. The VH comprises CDRH1, CDRH2 and CDRH3, the amino acid sequence of CDRH1 is SEQ ID No. 82, the amino acid sequence of CDRH2 is SEQ ID No. 171 and the amino acid sequence of CDRH3 is SEQ ID No. 260, and wherein the VL comprises CDRL1, CDRL2 and CDRL3, the amino acid sequence of CDRL1 is SEQ ID No. 349, the amino acid sequence of CDRL2 is SEQ ID No. 438 and the amino acid sequence of CDRL3 is SEQ ID No. 527. B. The VH comprises CDRH1, CDRH2 and CDRH3, the amino acid sequence of the CDRH1 is SEQ ID NO: 56, the amino acid sequence of the CDRH2 is SEQ ID NO: 145, and the amino acid sequence of the CDRH3 is SEQ ID NO: 234, and wherein the VL comprises CDRL1, CDRL2 and CDRL3, the amino acid sequence of the CDRL1 is SEQ ID NO: 323, the amino acid sequence of the CDRL2 is SEQ ID NO: 412, and the amino acid sequence of the CDRL3 is SEQ ID NO: 501, or C. The VH comprises CDRH1, CDRH2 and CDRH3, the amino acid sequence of CDRH1 is SEQ ID No. 86, the amino acid sequence of CDRH2 is SEQ ID No. 175 and the amino acid sequence of CDRH3 is SEQ ID No. 264, and wherein the VL comprises CDRL1, CDRL2 and CDRL3, the amino acid sequence of CDRL1 is SEQ ID No. 353, the amino acid sequence of CDRL2 is SEQ ID No. 442 and the amino acid sequence of CDRL3 is SEQ ID No. 531.
2. 2. The antibody or antibody fragment thereof of claim 1, wherein: a. The VH comprises the amino acid sequence of SEQ ID NO. 616 and the VL comprises the amino acid sequence of SEQ ID NO. 705; b. The VH comprises the amino acid sequence of SEQ ID NO: 590 and the VL comprises the amino acid sequence of SEQ ID NO: 679, or C. The VH comprises the amino acid sequence of SEQ ID NO. 620 and the VL comprises the amino acid sequence of SEQ ID NO. 709.
3. 3. The antibody or antibody fragment thereof of claim 1 or 2, wherein the antibody is a monoclonal antibody, a bispecific antibody, a multispecific antibody, a grafted antibody, a humanized antibody, a synthetic antibody, a chimeric antibody, a single chain antibody, a Fab fragment, a F (ab') 2 fragment, an Fv fragment, a disulfide-linked Fv (sdFv), an intracellular antibody, or an ab antigen-binding fragment thereof.
4. 4. The antibody or antibody fragment thereof of claim 1 or 2, wherein the antibody or antibody fragment thereof is chimeric or humanized.
5. 5. The antibody or antibody fragment thereof of claim 1 or 2, wherein the antibody has an EC50 of less than 25 nM in a cAMP assay.
6. 6. The antibody or antibody fragment thereof of claim 1 or 2, wherein the antibody has an EC50 of less than 20 nM in a cAMP assay.
7. 7. The antibody or antibody fragment thereof of claim 1 or 2, wherein the antibody has an EC50 of less than 10nM in a cAMP assay.
8. 8. A pharmaceutical composition comprising the antibody or antibody fragment thereof of claim 1 or claim 2.
9. 9. An isolated nucleic acid encoding the antibody or antibody fragment thereof of claim 1 or claim 2.
10. 10. A vector comprising the isolated nucleic acid of claim 9.
11. 11. A cell comprising the isolated nucleic acid of claim 9 or the vector of claim 10.
12. 12. A cell expressing the antibody or antibody fragment thereof of claim 1 or claim 2.
13. 13. Use of the antibody or antibody fragment thereof of claim 1 or claim 2 in the manufacture of a medicament for treating cancer in a subject in need thereof.

Description

Antibodies that bind to adenosine A2A receptor and methods of using the same for treating cancer and neurological diseases Cross reference The present application claims the benefit of U.S. provisional patent application No. 62/945,818 filed on date 2019, 12, 9, which is incorporated herein by reference in its entirety. Background G-protein coupled receptors (GPCRs), such as adenosine receptors, are associated with a variety of diseases. Because of the problems with obtaining a suitable antigen, it has been difficult to produce antibodies to GPCRs because GPCRs are typically expressed at low levels in cells and are very unstable when purified. Thus, there is a need for improved agents for therapeutic intervention that target GPCRs. Incorporation by reference All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. Disclosure of Invention Provided herein are methods of producing a library of nucleic acids encoding adenosine A2A receptor antibodies or antibody fragments thereof, the methods comprising (a) providing a predetermined sequence encoding i.a first plurality of polynucleotides, wherein each polynucleotide of the first plurality encodes a variant sequence encoding CDR1 on a heavy chain, ii.a second plurality of polynucleotides, wherein each polynucleotide of the second plurality encodes a variant sequence encoding CDR2 on a heavy chain, iii.a third plurality of polynucleotides, wherein each polynucleotide of the third plurality encodes a variant sequence encoding CDR3 on a heavy chain, and (b) mixing the first plurality of polynucleotides, the second plurality of polynucleotides and the third plurality of polynucleotides form a nucleic acid library encoding the adenosine A2A receptor antibodies or antibody fragments thereof. Further provided herein are methods of generating a library of nucleic acids encoding an adenosine A2A receptor antibody or antibody fragment thereof, wherein the adenosine A2A receptor antibody or antibody fragment thereof is a single domain antibody. Further provided herein are methods of generating a library of nucleic acids encoding adenosine A2A receptor antibodies or antibody fragments thereof, wherein the single domain antibody comprises a heavy chain variable domain. Further provided herein are methods of generating a library of nucleic acids encoding adenosine A2A receptor antibodies or antibody fragments thereof, wherein the single domain antibodies are VHH antibodies. Further provided herein are methods of producing a nucleic acid library encoding adenosine A2A receptor antibodies or antibody fragments thereof, wherein the nucleic acid library comprises at least 50,000 variant sequences. Further provided herein are methods of producing a nucleic acid library encoding adenosine A2A receptor antibodies or antibody fragments thereof, wherein the nucleic acid library comprises at least 100,000 variant sequences. Further provided herein are methods of producing a nucleic acid library encoding adenosine A2A receptor antibodies or antibody fragments thereof, wherein the nucleic acid library comprises at least 10 5 different nucleic acids. Further provided herein are methods of producing a nucleic acid library encoding an adenosine A2A receptor antibody or antibody fragment thereof, wherein the nucleic acid library comprises at least one sequence encoding an adenosine A2A receptor antibody or antibody fragment which binds to the adenosine A2A receptor with a K D of less than 100 nM. Further provided herein are methods of producing a nucleic acid library encoding an adenosine A2A receptor antibody or antibody fragment thereof, wherein the nucleic acid library comprises at least one sequence encoding an adenosine A2A receptor antibody or antibody fragment which binds to the adenosine A2A receptor with a K D of less than 50 nM. Further provided herein are methods of producing a nucleic acid library encoding an adenosine A2A receptor antibody or antibody fragment thereof, wherein the nucleic acid library comprises at least one sequence encoding an adenosine A2A receptor antibody or antibody fragment which binds to the adenosine A2A receptor with a K D of less than 10 nM. further provided herein are methods of producing a nucleic acid library encoding adenosine A2A receptor antibodies or antibody fragments thereof, wherein the nucleic acid library comprises at least 500 variant sequences. Further provided herein are methods of producing a nucleic acid library encoding an adenosine A2A receptor antibody or antibody fragment thereof, wherein the nucleic acid library comprises at least five sequences encoding an adenosine A2A receptor antibody or antibody fragment which binds to the adenosine A2A receptor with a K D of less than 100 nM. further provided