Apelin Fusion Proteins and Uses Thereof

Patent metadata

Field	Value
Publication number	US-2016237130-A1
Application number	US-201615146730-A
Country	US
Kind code	A1
Filing date	May 4, 2016
Priority date	Mar 14, 2013
Publication date	Aug 18, 2016
Grant date	—

Abstract

The invention provides a fusion protein or polypeptide comprising an apelin peptide fused to a multimerizing component. The invention also provides a fusion protein or polypeptide comprising an apelin peptide fused to an Fc domain, a fragment of an Fc domain, or a variant of an Fc domain. Apelin Fc-fusion polypeptides are capable of binding to the apelin receptor (APLNR). Apelin Fc-fusion polypeptides are capable of activating the APLNR and have improved pharmacokinetic properties compared to apelin peptides that are not fused to an Fc or an Fc fragment. Apelin Fc-fusion polypeptides are useful in diseases and conditions related to cardiovascular function, diabetes, cancer, obesity and other apelin-related conditions.

First claim

1 - 48 . (canceled)

49 . A polypeptide comprising an apelin peptide fused to an Fc domain, a fragment of an Fc domain, or variant of an Fc domain, wherein the N-terminus of the apelin peptide is fused to the C-terminus of the Fc domain or fragment or variant thereof, optionally via a peptide linker.

50 . The polypeptide of claim 49 , wherein the apelin peptide is selected from the group consisting of apelin42-77 (apelin-36), apelin61-77 (apelin-17), apelin63-77 (apelin-15), apelin64-77 (apelin-14), apelin65-77 (apelin-13), apelin66-77 (apelin-12), apelin67-77 (apelin-11), apelin68-77 (apelin-10), apelin73-77 (apelin-5), apelin61-76 (apelin-K16P), apelin61-75 (apelin-K15M), apelin61-74 (apelin-K14P), and [Pyr 1 ]Apelin-13.

51 . The polypeptide of claim 50 , wherein the apelin peptide is apelin-13.

52 . The polypeptide of claim 49 , wherein the apelin peptide is a modified apelin peptide having an additional amino acid at its C-terminus.

53 . The polypeptide of claim 52 , wherein the modified apelin peptide is a modified apelin-13.

54 . The polypeptide of claim 53 , wherein the modified apelin peptide comprises the amino acid sequence of SEQ ID NO: 42.

55 . The polypeptide of claim 53 , wherein the modified apelin peptide comprises the amino acid sequence of SEQ ID NO: 43.

56 . The polypeptide of claim 53 , wherein the modified apelin peptide comprises the amino acid sequence of SEQ ID NO: 44.

57 . The polypeptide of claim 49 , wherein the apelin peptide is fused to the Fc domain, or fragment thereof, via one or more peptide linkers.

58 . The polypeptide of claim 57 , wherein the apelin peptide is fused to the Fc domain, or fragment thereof, via one or more Gly-Ser linkers.

59 . The polypeptide of claim 49 , wherein the Fc domain is selected from the group consisting of IgG1 CH2 domain and IgG1 CH3 domain; IgG4 CH2 domain and IgG4 CH3 domain; IgG1 CH2 domain and IgG4 CH3 domain; and IgG4 CH2 domain and an IgG1 CH3 domain.

60 . The polypeptide of claim 49 , wherein the Fc domain further comprises an IgG hinge domain, or a fragment or variant thereof.

61 . The polypeptide of claim 60 , wherein the IgG hinge domain comprises SEQ ID NO: 14, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO:21, or SEQ ID NO: 22.

62 . The polypeptide of claim 49 , wherein the polypeptide is an APLNR agonist that exhibits an EC50 of less than about 10 nM, or less than about 1 nM when measured in an in vitro APLNR activation assay.

63 . The polypeptide of claim 49 , wherein the polypeptide has an in vivo half-life of at least about 1 hour, or at least about 2, 3, 4, 5, 6, 7, 8, 9, or 10 hours.

64 . A polypeptide comprising the amino acid sequence of SEQ ID NO: 39.

65 . A polypeptide comprising the amino acid sequence of SEQ ID NO: 40.

66 . A polypeptide comprising the amino acid sequence of SEQ ID NO: 41.

67 . A composition comprising the polypeptide of claim 49 and at least one pharmaceutically acceptable carrier or diluent.

68 . A nucleic acid molecule encoding the polypeptide of claim 49 .

69 . A vector comprising the nucleic acid molecule of claim 68 .

70 . A cell comprising the nucleic acid molecule of claim 68 .

71 . A cell comprising the vector of claim 69 .

72 . The cell of claim 70 , wherein the nucleic acid is stably integrated into the genome of the cell.

73 . The cell of claim 72 , wherein the cell is a eukaryotic cell.

74 . The cell of claim 73 , wherein the host cell is selected from the group consisting of CHO, COS, retinal cell, Vero, CV1, 293, MDCK, HaK, BHK, HeLa, HepG2, WI38, MRC 5, Colo25, HB 8065, HL-60, Jurkat, Daudi, A431 (epidermal), CV-1, U937, 3T3, L cell, C127 cell, SP2/0, NS-0, MMT cell, and tumor cell.

75 . The cell of claim 74 , wherein the cell is an animal cell.

76 . The cell of claim 75 , wherein the cell is a mammalian cell.

77 . The cell of claim 76 , wherein the cell is a CHO cell.

78 . The cell of claim 77 , wherein the cell is a CHO-K1 cell.

Assignees

• Regeneron Pharma

Inventors

• Stevis Panayiotis
• Murphy Andrew

Classifications

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P35/04

  specific for metastasis · CPC title

• A61P9/04

  Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure · CPC title