Ultralong complementarity determining regions and uses thereof

• US 10,774,132 B2
• Filed: 01/09/2013
• Issued: 09/15/2020
• Est. Priority Date: 01/09/2012
• Status: Active Grant

• Current Assignee: The Scripps Research Institute
• Original Assignee: The Scripps Research Institute
• Inventors: Smider, Vaughn, Bazirgan, Omar A., Mao, Hongyuan Helen, Schultz, Peter, Wang, Feng, Zhang, Yong
• Primary Examiner(s): Kemmerer, Elizabeth C.
• Assistant Examiner(s): DeBerry, Regina M
• Application Number:
  US13/737,910
• Publication Number:
  US 20140086871A1
• Time in Patent Office: 2,806 Days
• Field of Search: 5303873, 530351, 5303871, 4241341, 4241301, 4241331
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/53 Colony-stimulating factor [CSF]
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/10 from RNA viruses
• CPC Class Codes: C07K 16/46 Hybrid immunoglobulins hybrids of an immunoglobulin with a peptide not being an immunoglobulin C07K19/00
• CPC Class Codes: C07K 2317/20 characterized by taxonomic origin
• CPC Class Codes: C07K 2317/50 characterized by immunoglobulin fragments
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/569 Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/94 Stability, e.g. half-life, pH, temperature or enzyme-resistance
• CPC Class Codes: C07K 2318/00 Antibody mimetics or scaffolds
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 2319/50 containing protease site
• CPC Class Codes: C07K 2319/55 containing a fusion with a toxin, e.g. diphteria toxin

Abstract

Disclosed herein are immunoglobulin constructs comprising at least one immunoglobulin domain or fragment thereof; and a therapeutic polypeptide or derivative or variant thereof attached to or inserted into said immunoglobulin domain. Also provided are immunoglobulin constructs comprising a mammalian immunoglobulin heavy chain comprising at least a portion of a knob domain in the complementarity-determining region 3 (CDR3H) or fragment thereof; and a therapeutic polypeptide attached to or inserted into said knob domain of the CDR3H. Also provided are immunoglobulin constructs comprising a mammalian immunoglobulin heavy chain comprising at least a portion of a stalk domain in the complementarity-determining region 3 (CDR3H) or fragment thereof; and a therapeutic polypeptide attached to or inserted into said stalk domain of the CDR3H. Also described herein are methods and compositions comprising the immunoglobulin constructs described herein for treatment and prevention of a disease or condition in a subject.

Claims

1. A recombinant antibody comprising:

a modified antibody variable domain;

a therapeutic polypeptide; and

a stalk domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

157, 158, 159, 160, 192, 235, 236, 250, 259, 264, 285, and 289;

wherein the therapeutic polypeptide comprises one or more of;

a growth colony stimulating factor (GCSF), granulocyte macrophage colony-stimulating factor (GMCSF), fibroblast growth factor21 (FGF21), beta-interferon, exendin-4, glucagon-like peptide 1 (GLP-1), somatostatin, erythropoietin, Mokatoxin-1 (Moka1), VM-24, protoxin2, and a sequence comprising one or more sequences from SEQ ID NOS;

317-332;

wherein the therapeutic polypeptide is inserted into a complementarity-determining region of the antibody variable domain, wherein the therapeutic polypeptide is connected to the antibody variable domain by the stalk domain; and

wherein the therapeutic polypeptide within the recombinant antibody is functional to treat or ameliorate a disease, disorder or condition when the recombinant antibody is administered to a subject in need thereof.

2. The recombinant antibody of claim 1, wherein the antibody is selected from a chimeric antibody, a human engineered antibody, and a humanized antibody.

3. The recombinant antibody of claim 1, wherein the antibody is selected from a bovine antibody, a bovinized antibody, a bovine engineered antibody, and a fully bovine antibody.

4. The recombinant antibody of claim 1, wherein the therapeutic peptide comprises the growth colony stimulating factor (GCSF), granulocyte macrophage colony-stimulating factor (GMCSF), or fibroblast growth factor 21 (FGF21), or a combination thereof.

5. The recombinant antibody of claim 4, wherein the GCSF is selected from a bovine GCSF and a human GCSF.

6. The recombinant antibody of claim 1, wherein the therapeutic polypeptide comprises the beta-interferon, exendin-4, glucagon-like peptide 1 (GLP-1), somatostatin, erythropoietin, Mokatoxin-1 (Moka1), VM-24, or protoxin2, or a combination thereof.

7. The recombinant antibody of claim 1, wherein the therapeutic polypeptide comprises one or more of the sequences from SEQ ID NOS:

317-332.

8. The recombinant antibody of claim 1, comprising a linker sequence.

9. The recombinant antibody of claim 8, wherein the linker sequence is selected from (a) the sequence (GGGGS)_n and wherein n=1 to 5 (SEQ ID NO:

499);

(b) the sequence GGGSGGGGS (SEQ ID NO;

     337);

(c) the sequence GGGGSGGGS (SEQ ID NO;

     338); and

(d) a combination of (a)-(c).

10. The recombinant antibody of claim 1, further comprising one or more cleavage sites.

11. The recombinant antibody of claim 10, wherein the one or more cleavage sites comprise a recognition site for a protease.

12. The recombinant antibody of claim 10, wherein the one or more cleavage sites is located between the antibody variable domain and the therapeutic polypeptide.

13. A pharmaceutical composition comprising the recombinant antibody of claim 1.

14. The recombinant antibody of claim 1, wherein the recombinant antibody comprises an amino acid sequence selected the group consisting of SEQ ID NOs:

25-37.

15. The recombinant antibody of claim 1, wherein the variable domain is part of an antibody selected from the group consisting of an IgA, an IgD, an IgE, an IgG, an IgM, a Fab, a Fab′

, a F(ab′

)2, a single-chain Fv (scFv), a Fv, a dsFv, a diabody, a (ds Fv)2), a single chain antibody, a minibody, a flex minibody, and a Fd.