Claims for Patent: 9,879,077

Summary for Patent: 9,879,077

Title:	Anti-serum albumin Fab-effector moiety fusion construct, and a method of preparing the construct
Abstract:	The present invention relates to antigen-binding fragment (Fab) and a Fab-effector fusion protein or (poly)peptide comprising thereof. The Fab of the present invention specifically binds to serum albumin and thereby has extended in vivo half-life. The Fab of the present invention is characterized by not having cysteine residues that are responsible for the interchain disulfide bond in C.sub.H1 domain and C.sub..kappa.L domain as well. The Fab-effector fusion protein or (poly)peptide of the present invention can be produced in periplasm of E. coli with high yield, and has increased in vivo half-life. Further, the present invention provides E. coli strain which produces various kinds of Fab-effector fusion proteins or (poly)peptides, and a pharmaceutical composition comprising the fab-effector fusion proteins or (poly)peptides.
Inventor(s):	Cha; Sanghoon (Gangwon-Do, KR)
Assignee:	AprilBio Co., Ltd. (Chuncheon, Gangwon-do, KR)
Application Number:	15/056,299
Patent Claims:	1. An antigen binding fragment (Fab) to human serum albumin (SA), wherein the Fab comprises, (a) a heavy chain variable domain (V.sub.H domain) having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6; and (b) a light chain variable domain (V.sub.L domain) having an amino acid sequence selected from the group consisting of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, wherein the Fab binds specifically to human serum albumin. 2. An antigen binding fragment (Fab) binding to human serum albumin (SA), wherein the Fab comprises, (a) the amino acid sequences of SEQ ID NOS: 13 (CDR1), 14 (CDR2) and 15 (CDR3) determining the CDRs of the V.sub.H domain; and (b) the amino acid sequences of SEQ ID NOS: 16 (CDR1), 17 (CDR2) and 18 (CDR3) determining the CDRs of the V.sub.L domain. 3. The Fab according to claim 2, wherein the V.sub.H domain is bound to the heavy chain constant 1 domain (C.sub.H1 domain), and V.sub.L domain is bound to the light chain constant domain (C.sub..kappa.L domain). 4. The Fab according to claim 3, wherein the V.sub.H domain has the amino acid sequence of SEQ ID NO: 6 and the V.sub.L domain has the amino acid sequence of SEQ ID NO: 12. 5. The Fab according to claim 1 or claim 2, further comprising a C.sub.H1 domain and a C.sub..kappa.L domain, wherein at least one of the amino acid cysteines of the C.sub.H1 domain and the C.sub..kappa.L domain is deleted or substituted with a different amino acid residue, including serine, except for cysteine. 6. The Fab according to claim 5, wherein the amino acid cysteine of C.sub.H1 domain is the 233.sup.th amino acid starting from the N-terminus of the C.sub.H1 domain, and the cysteine of C.sub..kappa.L domain is the 214.sup.th amino acid starting from the N-terminus of the C.sub..kappa.L domain. 7. A fusion construct of the antigen binding fragment (Fab) of claim 1 or claim 2 and a bioactive effector moiety; wherein the bioactive effector moiety is a protein or a poly peptide; and wherein the Fab and the bioactive effector moiety are covalently linked by genetic fusion. 8. The fusion construct according to claim 7, wherein the Fab and the bioactive effector moiety are covalently linked by genetic fusion using a peptide linker of 1 to 20 amino acids. 9. The fusion construct according to claim 7, wherein the bioactive effector moiety is one selected from the group consisting of hormone, cytokine, enzyme, antibody, growth factor, transcription factor, blood factor, vaccine, ligand protein, and receptor. 10. The fusion construct according to claim 7, wherein the bioactive effector moiety is one selected from the group consisting of human growth hormone (hGH), growth hormone releasing hormone (GHRH), growth hormone releasing peptide, interferons (IFNs), interferon receptors, colony stimulating factors (CSFs), glucagon-like peptides, G-protein-coupled receptors, interleukins, interleukin receptors, enzymes, interleukin binding proteins, cytokine binding proteins, macrophage activating factor, macrophage peptide, B cell factor, T cell factor, protein A, cell necrosis glycoproteins, lymphotoxin, tumor necrosis factor, tumor suppressors, metastasis growth factor, alpha-1 antitrypsin, albumin, alpha-lactalbumin, apolipoprotein-E, erythropoietin, highly glycosylated erythropoietin, angiopoietins, hemoglobin, thrombin, thrombin receptor activating peptide, thrombomodulin, factor VII, factor VIIa, factor VIII, factor IX, factor XIII, plasminogen activating factor, fibrin-binding peptide, urokinase, streptokinase, hirudin, protein C, C-reactive protein, renin inhibitor, collagenase inhibitor, superoxide dismutase, leptin, platelet-derived growth factor, epithelial growth factor, epidermal growth factor, angiostatin, angiotensin, bone growth factor, bone stimulating protein, calcitonin, insulin, atriopeptin, cartilage inducing factor, elcatonin, connective tissue activating factor, tissue factor pathway inhibitor, follicle stimulating hormone, luteinizing hormone, luteinizing hormone releasing hormone, nerve growth factors, parathyroid hormone, relaxin, secretin, somatomedin, insulin-like growth factor, adrenocortical hormone, glucagon, cholecystokinin, pancreatic polypeptide, gastrin releasing peptide, corticotropin releasing factor, thyroid stimulating hormone, autotaxin, lactoferrin, myostatin, receptors, receptor antagonists, cell surface antigens, virus derived vaccine antigens, monoclonal antibodies, polyclonal antibodies, and antibody fragments. 11. The fusion construct according to claim 10, wherein the bioactive effector moiety is hGH, granulocyte colony stimulating factor (GCSF), or IFNs. 12. The fusion construct according to claim 7, wherein the molar ratio of the bioactive poly peptide or protein to the Fab is between 1:1 and 10:1. 13. The fusion construct according to claim 7, wherein the molar ratio of the bioactive polypeptide or protein to the Fab is between 1:1 and 4:1. 14. An expression vector comprising: (a) promoter; (b) a first nucleic acid sequence encoding the Fab of claim 1 or claim 2; and (c) a second nucleic acid sequence encoding a bioactive poly peptide or protein and optionally a linker, wherein the promoter, the first nucleic acid sequence and the second nucleic acid sequence are operably linked. 15. A host cell comprising the expression vector of claim 14. 16. The host cell according to claim 15, in which the host cell is E. coli. 17. The host cell according to claim 16, in which the host cell is SUPEX5 (KCTC 12657BP). 18. A pharmaceutical composition comprising the fusion construct of claim 7, and a pharmaceutically acceptable excipient.

Details for Patent 9,879,077

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Microbix Biosystems Inc.	KINLYTIC	urokinase	For Injection	021846	01/16/1978		2033-08-30
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965		2033-08-30
Nps Pharmaceuticals, Inc.	NATPARA	parathyroid hormone	For Injection	125511	01/23/2015		2033-08-30