Apelin Fusion Proteins and Uses Thereof

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.

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, and wherein the apelin peptide is selected from the group consisting of apelin61-77 (apelin-17), apelin63-77 (apelin-15), apelin64-77 (apelin-14), 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), apelin65-77 (apelin-F13A), apelin65-76, apelin65-75, apelin66-76, apelin67-76, apelin66-75, apelin67-75, and [Pyr 1 ]Apelin-13. 
     
     
         50 . The polypeptide of  claim 49 , wherein the apelin peptide is selected from the group consisting of apelin65-77 (apelin-F13A), apelin65-76, apelin65-75, apelin66-76, apelin67-76, apelin66-75, apelin67-75, apelin66-77 (apelin-12), and apelin67-77 (apelin-11). 
     
     
         51 . The polypeptide of  claim 50 , wherein the apelin peptide is apelin65-75. 
     
     
         52 . The polypeptide of  claim 50 , wherein the apelin peptide is apelin66-77 (apelin-12). 
     
     
         53 . The polypeptide of  claim 50 , wherein the apelin peptide is apelin66-76. 
     
     
         54 . The polypeptide of  claim 50 , wherein the apelin peptide is apelin67-76. 
     
     
         55 . The polypeptide of  claim 50 , wherein the apelin peptide is apelin66-75. 
     
     
         56 . The polypeptide of  claim 49 , wherein the apelin peptide is fused to the Fc domain, or fragment thereof, via one or more peptide linkers, optionally wherein the one or more peptide linkers comprise one or more Gly-Ser linkers. 
     
     
         57 . 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, optionally wherein the Fc domain further comprises an IgG hinge domain or a fragment or variant thereof. 
     
     
         58 . The polypeptide of  claim 57 , wherein the Fc domain comprises an IgG hinge domain comprising SEQ ID NO: 14, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO:21, or SEQ ID NO: 22. 
     
     
         59 . The polypeptide of  claim 49 , wherein the polypeptide (i) 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, and/or (ii) 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. 
     
     
         60 . A composition comprising the polypeptide of  claim 49  and at least one pharmaceutically acceptable carrier or diluent. 
     
     
         61 . A nucleic acid molecule encoding the polypeptide of  claim 49 . 
     
     
         62 . A vector comprising the nucleic acid molecule of  claim 61 . 
     
     
         63 . A cell comprising the nucleic acid molecule of  claim 61 . 
     
     
         64 . The cell of  claim 63 , wherein the nucleic acid is stably integrated into the genome of the cell. 
     
     
         65 . The cell of  claim 63 , wherein the cell is a eukaryotic cell, optionally wherein the 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. 
     
     
         66 . The cell of  claim 63 , wherein the cell is an animal cell, optionally wherein the cell is a mammalian cell. 
     
     
         67 . The cell of  claim 66 , wherein the cell is a CHO cell. 
     
     
         68 . The cell of  claim 67 , wherein the cell is a CHO-K1 cell.