Peptide-peptidase inhibitor conjugates and methods of using same

Abstract

Peptide-peptidase inhibitor conjugate molecules are disclosed. These conjugate molecules are useful as agents for the treatment and prevention of metabolic and cardiovascular diseases, disorders, and conditions. Such diseases, conditions and disorders include, but are not limited to, hypertension, dyslipidemia, cardiovascular disease, eating disorders, insulin-resistance, obesity, and diabetes mellitus of any kind, and other diabetes-related disorders.

We claim: 
     
         1 . A peptide-peptidase inhibitor conjugate comprising a peptide covalently linked to a peptidase inhibitor. 
     
     
         2 . The conjugate according to  claim 1 , wherein the peptide is selected from the group consisting of a GLP-1 peptide, a GIP peptide, a GLP-2 peptide, an exendin, a PYY peptide, an amylin peptide, a calcitonin, a leptin and analogs or hybrids thereof. 
     
     
         3 . The conjugate according to  claim 1 , wherein the peptidase inhibitor is selected from the group consisting of a vasopeptidase inhibitor, an angiotensin converting enzyme inhibitor, an endothelial converting enzyme inhibitor, a human endopeptidase inhibitor, a dipeptidyl peptidase IV (DPP-IV) inhibitor an aminopeptidase inhibitor, a cysteine protease inhibitor, a serine protease inhibitor, and a carboxypeptidase inhibitor. 
     
     
         4 . The conjugate according to  claim 2 , wherein the peptide is GLP-1 or an analog of GLP-1 comprising a sequence as set forth in any one of SEQ ID NOs: 41-59. 
     
     
         5 . The conjugate according to  claim 2 , wherein the peptide is GIP or an analog of GIP comprising a sequence as set forth in any one of SEQ ID NOs: 74-88 or 293-295. 
     
     
         6 . The conjugate according to  claim 1 , wherein the peptide-peptidase inhibitor conjugate is synthesized chemically or recombinantly. 
     
     
         7 . The conjugate according to  claim 1 , wherein the plasma half-life of the conjugate is greater than the plasma half-life of the peptide not conjugated to the peptidase inhibitor. 
     
     
         8 . The conjugate according to  claim 1 , wherein the conjugate is selected from the group consisting of Lys 30 N-ε-(Ado-(γ-L-Glu-2-(S)-cyano pyrrolidyl)) GIP(1-30), GIP(1-30) Ado-Lys-N-ε-(2-(S)-cyano pyrrolidyl), GIP(1-30) Ado-Ado-Lys-N-ε-(2-(S)-cyano pyrrolidyl), [Leu 14 ] GIP(1-30) Ado-Lys-N-ε-(2-(S)-cyano pyrrolidyl), and N-(γ-L-Glu-2-(S)-cyano pyrrolidyl)-Ado-Ado) GIP(1-30). 
     
     
         9 . The conjugate according to  claim 2 , wherein the peptide is a GLP-1 hybrid or a GIP hybrid and the peptidase inhibitor is a dipeptidyl peptidase IV (DPP-IV) inhibitor. 
     
     
         10 . The conjugate according to  claim 10 , wherein the GIP hybrid comprises a GIP peptide linked to an exendin peptide or an amylin peptide. 
     
     
         11 . The conjugate according to  claim 11 , wherein the GIP hybrid comprises the sequence as set forth in any one of SEQ ID NOs: 1, 291, 292, or 233-246. 
     
     
         12 . The conjugate according to  claim 11 , wherein the GIP-hybrid-DPP-IV inhibitor conjugate is selected from the group consisting of [D-Ala 2 ]GIP(1-30)-exendin-4(31-39)-(Lys-ε-NH-(Aun-Aun-γ-L-Glu-(2-(S)-cyano pyrrolidyl))))), GIP(1-30)-exendin-4(31-39)-Ado-Lys-N-ε-(2-(S)-cyano pyrrolidyl), GIP(1-30)-exendin-4(31-39)-Ado-Ado-Lys-N-ε-(2-(S)-cyano pyrrolidyl), [Leu 14 ] GIP(1-30)-exendin-4(31-39)-Ado-Lys-N-ε-(2-(S)-cyano pyrrolidyl), [D-Ala 2 ]GIP(1-30)-exendin-4(31-39)-Ado-Lys-N-ε-(2-(S)-cyano pyrrolidyl), Lys 30 N-ε-(Ado-(γ-L-Glu-2-(S)-cyano pyrrolidyl)) GIP(1-30)-exendin-4(31-39), [D-Ala 2 Lys 16 (ε-NH-(Aun-Aun-(γ-L-Glu-(2-(S)-cyano pyrrolidyl))))] GIP(1-30)-exendin-4(31-39), N-(γ-L-Glu-2-(S)-cyano pyrrolidyl)-Ado-Ado) GIP(1-30)-exendin-4(31-39), and [D-Ala 2 Lys(ε-NH-(Aun-Aun-(γ-L-Glu-(2-(S)-cyano pyrrolidyl))))] GIP(1-30)-exendin-4(31-39). 
     
     
         13 . A pharmaceutical composition comprising the conjugate of  claim 1 . 
     
     
         14 . A method for treating a metabolic disease in a subject in need thereof, the method comprising administering a therapeutically effective amount of the conjugate of  claim 1 . 
     
     
         15 . A method for decreasing degradation of a peptidase-sensitive peptide in a subject, the method comprising administering a composition comprising the peptide-peptidase inhibitor conjugate compound according to  claim 1  to a subject in an amount effective to decrease degradation of a peptidase-sensitive peptide. 
     
     
         16 . A method for increasing plasma concentration of a biologically active, peptidase-sensitive peptide in a subject, the method comprising administering a composition comprising the peptide-peptidase inhibitor according to  claim 1  to a subject in an amount effective to increase plasma half-life of a biologically active, peptidase-sensitive peptide.