IMMUNOGLOBULIN FUSION PROTEINS AND COMPOSITIONS THEREOF
Disclosed herein are immunoglobulin fusion proteins that have a first antibody region attached to an extender fusion region. The extender fusion region contains a therapeutic agent and a beta strand secondary structure. The extender fusion region may contain 7 or fewer consecutive amino acids based on or derived from an ultralong CDR3. Alternatively, the extender fusion region contains a rigid stalk protein structure, but does not contain an amino acid sequence based on or derived from an ultralong CDR3. The extender fusion region may also have one or more linkers or proteolytic cleavage sites. The immunoglobulin fusion proteins may have additional therapeutic agents and extender fusion regions. Also disclosed herein are pharmaceutical compositions of immunoglobulin fusion proteins and methods for using the immunoglobulin fusion proteins for the treatment or prevention of a disease or condition in a subject.
1 .- 61 . (canceled)
62 . An immunoglobulin fusion protein comprising:
a) a first antibody region; and
b) a first extender fusion region comprising a first therapeutic agent attached to a first extender peptide,
wherein the first extender peptide comprises a first beta strand secondary structure region having a first beta strand secondary structure;
wherein the first extender fusion region does not contain more than 7 consecutive amino acids from an ultralong complementary determining region 3 heavy chain of SEQ ID NO: 248.
63 . The immunoglobulin fusion protein of claim 62 , wherein the first extender fusion region comprises a second extender peptide comprising a second beta strand secondary structure region having a second beta strand secondary structure, and wherein the first beta strand secondary structure and second beta strand secondary structure form a beta sheet.
64 . The immunoglobulin fusion protein of claim 62 , wherein the first extender fusion region replaces a portion of a light chain of the first antibody region or a portion of the heavy chain of the first antibody region.
65 . The immunoglobulin fusion protein of claim 62 , wherein the first antibody region comprises an antibody selected from an anti-Her2 antibody, trastuzumab, an anti-CD47 antibody, palivizumab, and antigen binding fragments thereof.
66 . The immunoglobulin fusion protein of claim 62 , wherein the first antibody region comprises a heavy chain, wherein the heavy chain is represented by an amino acid sequence that is at least 90% homologous to a sequence selected from SEQ ID NOS: 24-27, 29-33, 36-39, and 251-253, and a light chain, wherein the light chain is represented by an amino acid sequence that is at least 90% homologous to a sequence selected from SEQ ID NOS: 21-23, 28, 34, 35, 40, 248-250 and 278.
67 . The immunoglobulin fusion protein of claim 62 , wherein the first beta strand secondary structure region comprises an amino acid sequence that is at least 90% homologous to a sequence selected from SEQ ID NOs: 109-128, 305-308.
68 . The immunoglobulin fusion protein of claim 62 , wherein the first beta strand secondary structure region comprises an amino acid sequence of ETKKYQX n S (SEQ ID NO:110), wherein n=1-8 and X is selected from a basic amino acid, an acidic amino acid, a polar amino acid, and a charged amino acid.
69 . The immunoglobulin fusion protein of claim 63 , wherein the first beta strand secondary structure region comprises an amino acid sequence of ETKKYQX n S (SEQ ID NO: 306), wherein n=1-8 and X is selected from a basic amino acid, an acidic amino acid, a polar amino acid, and a charged amino acid, and wherein the second beta strand secondary structure region comprises an amino acid sequence of SX 1 TX 2 NX 3 (SEQ ID NO: 128), wherein X 1 , X 2 , and X 3 are independently selected from polar amino acids.
70 . The immunoglobulin fusion protein of claim 63 , wherein the second beta strand secondary structure region comprises an amino acid sequence of YX 1 YX 2 Y, and wherein X 1 and X 2 are independently selected from polar amino acids.
71 . The immunoglobulin fusion protein of claim 62 , wherein the immunoglobulin fusion protein comprises an amino acid sequence that is at least about 90% identical to an amino acid sequence of any one of SEQ ID NOs: 76-108, 260-277, 298, 300, 302, and 304.
72 . The immunoglobulin fusion protein of claim 62 , wherein the immunoglobulin fusion protein further comprises a second therapeutic agent.
73 . The immunoglobulin fusion protein of claim 72 , wherein the immunoglobulin fusion protein further comprises a second antibody region, wherein the second antibody region comprises a second extender fusion region attached to the second antibody region, and wherein the second extender fusion region comprises the second therapeutic agent and an extender peptide comprising
a) an amino acid sequence having an alpha helical secondary structure, or
b) an amino acid sequence having a beta strand secondary structure, or
c) an amino acid sequence having no regular secondary structure.
74 . The immunoglobulin fusion protein of claim 72 , further comprising a second antibody region, wherein the second therapeutic agent is directly attached to the second antibody region.
75 . The immunoglobulin fusion protein of claim 62 , wherein first therapeutic agent is selected from bGCSF, hGCSF, bGMCSF, hGMCSF, GDF11, interferon-beta, interferon-alpha, interleukin 11 (IL-11), exendin-4, GLP-1, relaxin, oxyntomodulin, leptin, betatrophin, bovine growth hormone (bGH), human growth hormone (hGH), parathyroid hormone, erythropoietin, Moka1, VM-24, Mamba1, angiopoeitin-like 3 (ANGPTL3), CVX15, a CXCR4 ligand, a neutrophil elastase inhibitor, and homologs thereof.
76 . The immunoglobulin fusion protein of claim 62 , wherein the first therapeutic agent is represented by an amino acid sequence that is at least 90% homologous to a sequence selected from any one of SEQ ID NOs: 200-235 and/or encoded by a nucleic acid sequence that is at least 90% homologous to a sequences selected from SEQ ID NOs: 167-199.
77 . The immunoglobulin fusion protein of claim 62 , wherein the first antibody region comprises a BVK antibody or antigen binding fragment thereof and the first therapeutic agent comprises a Moka1 peptide, and wherein the first extender fusion region is grafted into a heavy chain of the BVK antibody.
78 . The immunoglobulin fusion protein of claim 77 , wherein the Moka1 peptide is represented by an amino acid sequence that is at least about 90% homologous to SEQ ID NO. 202.
79 . The immunoglobulin fusion protein of claim 77 , wherein the heavy chain of the BVK antibody is encoded by a nucleotide sequence that is at least 90% homologous to SEQ ID NO. 1.
80 . The immunoglobulin fusion protein of claim 77 , wherein the first beta strand secondary structure is represented by an amino acid sequence selected from SEQ ID NOS: 111, 119, and combinations thereof.
81 . The immunoglobulin fusion protein of claim 77 , wherein:
a) the BVK antibody comprises a heavy chain encoded by a nucleotide sequence of SEQ ID NO:1, and a light chain encoded by a nucleotide sequence of SEQ ID NO: 14;
b) the Moka1 peptide is represented by an amino acid sequence of SEQ ID NO: 202;
c) the first beta strand secondary structure is represented by an amino acid sequence of SEQ ID NO: 111; and
d) a second beta strand secondary structure is represented by an amino acid sequence of SEQ ID NO: 119,
wherein the Moka1 peptide is grafted into the heavy chain of the BVK antibody.