Natriuretic peptide conjugate using carrier substance

Abstract

The present invention relates to an Natriuretic peptide conjugate having improved in-vivo duration of efficacy and stability, comprising an Natriuretic peptide, a non-peptidyl polymer and a carrier substance, which are covalently linked to each other, and a use of the same. The Natriuretic peptide conjugate of the present invention has the in-vivo activity which is maintained relatively high, and has remarkably increased blood half-life, and thus it can be desirably employed in the development of long-acting formulations of various peptide drugs.

1 . A Natriuretic peptide conjugate, comprising a Natriuretic peptide and an immunoglobulin Fc region, which are linked by a non-peptidyl polymer, wherein the non-peptidyl polymer is selected from the group consisting of polyethylene glycol, polypropylene glycol, copolymers of ethylene glycol and propylene glycol, polyoxyethylated polyols, polyvinyl alcohol, polysaccharides, dextran, polyvinyl ethyl ether, biodegradable polymers, lipid polymers, chitins, hyaluronic acid, and combinations thereof.  
     
     
         2 . The Natriuretic peptide conjugate according to  claim 1 , wherein the Natriuretic peptide is selected from the group consisting of Atrial Natriuretic Peptide(ANP), Brain Natriuretic peptide(BNP), C-type Natriuretic peptide(CNP), Dendroaspis Natriuretic peptide(DNP), and a derivative, a fragment and a variant thereof.  
     
     
         3 . The Natriuretic peptide conjugate according to  claim 2 , wherein the Natriuretic peptide is native BNP  
     
     
         4 . The Natriuretic peptide conjugate according to any one of  claims 1  to  3 , wherein the non-peptidyl polymer has both ends, each binding to an amine group or a thiol group of the immunoglobulin Fc region and native BNP.  
     
     
         5 . The Natriuretic peptide conjugate according to  claim 4 , wherein the non-peptidyl polymer has both ends, each binding to an N-terminus of the immunoglobulin Fc region, and native BNP.  
     
     
         6 . The Natriuretic peptide conjugate according to  claim 1 , wherein the immunoglobulin Fc region is deglycosylated.  
     
     
         7 . The Natriuretic peptide conjugate according to  claim 1 , wherein the immunoglobulin Fc region is composed of one to four domains selected from the group consisting of CH1, CH2, CH3 and CH4 domains.  
     
     
         8 . The Natriuretic peptide conjugate according to  claim 7 , wherein the immunoglobulin Fc region further includes a hinge region.  
     
     
         9 . The Natriuretic peptide conjugate according to  claim 1 , wherein the immunoglobulin Fc region is an Fc region derived from an immunoglobulin selected from the group consisting of IgG, IgA, IgD, IgE, or IgM.  
     
     
         10 . The Natriuretic peptide conjugate according to  claim 9 , wherein each domain of the immunoglobulin Fc region is a domain hybrid of a different origin derived from an immunoglobulin selected from the group consisting of IgG, IgA, IgD, IgE, and IgM.  
     
     
         11 . The Natriuretic peptide conjugate according to  claim 9 , wherein the immunoglobulin Fc region is a dimer or a multimer (a combination of immunoglobulin Fc) composed of single-chain immunoglobulin of the same origin.  
     
     
         12 . The Natriuretic peptide conjugate according to  claim 9 , wherein the immunoglobulin Fc region is an IgG4 Fc region.  
     
     
         13 . The Natriuretic peptide conjugate according to  claim 12 , wherein the immunoglobulin Fc region is a human deglycosylated IgG4 Fc region.  
     
     
         14 . The Natriuretic peptide conjugate according to  claim 1 , wherein the reactive group of the non-peptidyl polymer is selected from the group consisting of an aldehyde group, a propione aldehyde group, a butyl aldehyde group, a maleimide group, and a succinimide derivative.  
     
     
         15 . The Natriuretic peptide conjugate according to  claim 14 , wherein the succinimide derivative is succinimidyl propionate, succinimidyl carboxymethyl, hydroxy succinimidyl, or succinimidyl carbonate.  
     
     
         16 . The Natriuretic peptide conjugate according to  claim 14 , wherein the non-peptidyl polymer has a reactive aldehyde group at both ends.  
     
     
         17 . The Natriuretic peptide conjugate according to  claim 1 , wherein the non-peptidyl polymer is polyethylene glycol.  
     
     
         18 . A method for preparing an Natriuretic peptide conjugate, comprising the steps of: 
 (1) Covalently linking a non-peptidyl polymer having a reactive group selected from the group consisting of aldehyde, maleimide, and succinimide derivatives at both ends thereof, with an amine or thiol group of a Natriuretic peptide;    (2) Isolating a conjugate comprising the Natriuretic peptide, in which the non-peptidyl polymer from the reaction mixture of (1) is linked covalently to N-terminus; and    (3) Covalently linking an immunoglobulin Fc region to the other end of the non-peptidyl polymer of the isolated conjugate to produce a peptide conjugate comprising the immunoglobulin Fc region and the Natriuretic peptide, which are linked to each end of the non-peptidyl polymer.    
     
     
         19 . The method for preparing the Natriuretic peptide conjugate according to  claim 18 , wherein the non-peptidyl polymer is polyethylene glycol.  
     
     
         20 . The method for preparing the Natriuretic peptide conjugate according to  claim 18 , wherein the Natriuretic peptide is native BNP.  
     
     
         21 . A pharmaceutical composition comprising the peptide conjugate of any one of  claims 1  to  17 .  
     
     
         22 . A method for treating acute or chronic congestive heart failure (CHF), hypertension, asthma, inflammation related disease, hyperlipidemia or impotence comprising administering the peptide conjugate of any one of  claims 1  to  17 , or the pharmaceutical composition of  claim 21.