Oxyntomodulin analogs

Abstract

Peptide analogs of oxyntomodulin (OXM, glucagon-37), which have been modified to be resistant to cleavage and inactivation by dipeptidyl peptidase IV (DPP-IV) and to increase in vivo half-life of the peptide analog while enabling the peptide analog to act as a dual GLP-1/glucagon receptor (GCGR) agonistm are described. The peptide analogs are useful for treatment of metabolic disorders such as diabetes and obesity.

1 . A peptide analog comprising the amino acid sequence
 HSQGTFTSDYSKYLDSRRAQDFVQWLMNTK (SEQ ID NO:109) in which   (A) the second amino acid from the N-terminus of the peptide is substituted with an amino acid that renders the peptide resistant to cleavage and inactivation by dipeptidyl peptidase IV;   (B) the peptide includes a lipid or cholesterol moiety covalently linked to the peptide by a spacer comprising one or more gamma-glutamic acid residues;   (C) the peptide optionally includes one to three amino acid substitutions in addition to the substitution at position 2; and   (D) the peptide optionally includes a protecting group that, if present, is joined to the C-terminal carboxy group of the peptide;   wherein the peptide analog has a pI of less than 6.0 and is a dual GLP-1 receptor agonist and glucagon receptor agonist, and pharmaceutically acceptable salts thereof.   
     
     
         2 . The peptide analog of  claim 1 , wherein the amino acid substituted for the second amino acid from the N-terminus is selected from the group consisting of D-serine, α-aminoisobutyric acid, and 1-amino-1-cyclobutane carboxylic acid. 
     
     
         3 . The peptide analog of  claim 1 , wherein the peptide analog includes a cholesterol moiety covalently linked to the thiol group of a cysteine residue that is covalently linked to the s-amino group of the lysine residue at the C-terminus of the peptide analog by a spacer comprising one or more gamma glutamic acid residues. 
     
     
         4 . The peptide analog of  claim 3 , wherein the cholesterol moiety is covalently linked to the thiol group by a hydrophilic linker. 
     
     
         5 . The peptide analog of  claim 4 , wherein the hydrophilic linker is an ethoxy polymer that includes one to twelve ethoxy units. 
     
     
         6 . The peptide analog of  claim 4 , wherein the hydrophilic linker is an ethoxy polymer that includes four ethoxy units. 
     
     
         7 . The peptide analog of  claim 1 , wherein the peptide analog includes a lipid moiety covalently linked to the ε-amino group of a lysine residue. 
     
     
         8 . The peptide analog of  claim 7 , wherein the lipid moiety is a palmitoyl group. 
     
     
         9 . The peptide analog of  claim 7 , wherein the lipid moiety is covalently linked to the ε-amino group of the lysine residue by one or more gamma-glutamic acid residues. 
     
     
         10 . The peptide analog of  claim 7 , wherein the lipid moiety is covalently linked to the ε-amino group of the lysine residue at the C-terminus by one or more gamma-glutamic acid residues. 
     
     
         11 . The peptide analog of  claim 7 , wherein the lipid moiety is covalently linked to the ε-amino group of the lysine residue by one or more gamma-glutamic acid residues and the lysine residue linked to the lysine residue at the C-terminus by one or more gamma-glutamic acid residues. 
     
     
         12 . The peptide analog of  claim 1 , wherein the peptide analog further includes one or more amino acid substitutions at amino acid positions selected from the group consisting of positions 10, 12, 16, 17, 18, and 27. 
     
     
         13 . The peptide analog of  claim 1 , wherein the peptide analog further includes one or more amino acid substitutions selected from the group consisting of lysine for the tyrosine at position 10, serine for the lysine at position 12, glutamic acid or a-aminoisobutyric acid for the serine at position 16, glutamic acid for the arginine at position 17, alanine for the arginine at position 18, lysine for the glutamine at position 20, and norleucine or O-methyl-L-homoserine for the methionine at position 27. 
     
     
         14 . The peptide analog of  claim 13 , wherein tyrosine at position 10 is replaced with a lysine and the lipid moiety is covalently linked to the ε-amino group of the lysine residue by one or more gamma-glutamic acid residues. 
     
     
         15 . The peptide analog of  claim 13 , wherein glutamine at position 20 is replaced with lysine and the lipid moiety is covalently linked to the ε-amino group of the lysine by one or more gamma-glutamic acid residues. 
     
     
         16 . The peptide analog of  claim 14 , wherein the peptide analog further includes one or more gamma-glutamic acid residues covalently linked to the C-terminus. 
     
     
         17 . The peptide analog of  claim 15 , wherein the peptide analog further includes one or more gamma-glutamic acid residues covalently linked to the C-terminus. 
     
     
         18 . (canceled) 
     
     
         19 . A peptide analog comprising the structure
   Z 1 -P-M-Z 2      wherein P is a peptide having the amino acid sequence   
       
         
           
                 
               
                   (SEQ ID NO: 110) 
                 
                   HX 1 QGTFTSDX 2 SX 3 YLDX 4 X 5 X 6 AX 7 DFVQWLX 8 NTKX 9 X 10   
                 
             
                
                
               
            
           
         
         wherein X 1  is a D-serine, α-aminoisobutyric acid (aib), 1-Amino-1-cyclobutane carboxylic acid (Acb) residue, 1-Amino-1-cyclohexane carboxylic acid (Acx); alpha-aminobutyric acid (Abu); D-alpha-aminobutyric acid (D-Abu); Aminovaleric acid Nva); beta-cyclopropyl-alanine (Cpa); propargylglycine (Prg); Allylglycine (Alg); 2-Amino-2-cyclohexyl-propanoic acid (2-Cha); D-tertbutylglycine (D-tbg); Vinylglycine (Vg); 1-Amino-1-cyclopropane carboxylic acid (Acp); or 1-Amino-1-cyclopentane carboxylic acid (Acpe) residue; 
         X 2  is a tyrosine (Y) or lysine (K) residue; 
         X 3  is serine (S) or lysine (K) residue; 
         X 4  is serine (S), α-aminoisobutyric acid (aib), or glutamic acid (E) residue; 
         X 5  is an arginine (R) or glutamic acid (E) residue; 
         X 6  is an arginine (R) or alanine (A) residue; 
         X 7  is a glutamine (Q) or lysine (K) residue; 
         X 8  is a methionine (M), norleucine (Nle), methionine sulfoxide (m), or O-methyl-L-homoserine (o) residue; 
         X 9  is a gamma glutamic acid (γGlu) residue; 
         X 10  is a gamma glutamic acid (γGlu) residue or absent; 
         Z 1  is an optionally present protecting group that, if present, is joined to the N-terminal amino group, 
         M is (i) a cysteine residue covalently linked to a cholesterol moiety by a hydrophilic linker, (ii) a lysine residue covalently linked to a lipid moiety by a spacer comprising one or more gamma glutamic acid residues, or (iii) a lipid moiety, wherein M is covalently linked to C-terminal or internal amino acid of P by a spacer comprising one or more gamma-glutamic acid residues; and 
         Z 2  is an optionally present protecting group that, if present, is joined to the C-terminal carboxy group; and pharmaceutically acceptable salts thereof, 
         wherein the peptide analog or salt thereof has a pI of less than 6.0 and is a dual GLP-1 receptor agonist and glucagon receptor agonist. 
       
     
     
         20 . The peptide analog of  claim 19 , wherein M is a cysteine residue covalently linked to a cholesterol moiety with a hydrophilic linker and the cysteine residue is linked to the C-terminus of P. 
     
     
         21 . The peptide analog of  claim 19 , wherein M is a lysine residue covalently linked to a lipid moiety by a spacer comprising one or more gamma glutamic acid residues and the lysine residue is linked to the C-terminus of P. 
     
     
         22 . The peptide analog of  claim 19 , wherein M is a lysine residue covalently linked to a lipid moiety by a spacer comprising one or more gamma glutamic acid residues and the lysine residue is at position X 2  or X 7  of P. 
     
     
         23 . The peptide analog of  claim 19 , wherein the hydrophilic linker is an ethoxy polymer that includes one to twenty-four ethoxy units. 
     
     
         24 . The peptide analog of  claim 19 , wherein the hydrophilic linker is an ethoxy polymer that includes four ethoxy units. 
     
     
         25 . The peptide analog of  claim 19 , wherein the lipid moiety is a palmitoyl moiety. 
     
     
         26 . The peptide analog of  claim 19 , wherein the peptide analog is OXM317 (SEQ ID NO:60); OXM318 (SEQ ID NO:61); OXM319 (SEQ ID NO:62); OXM323 (SEQ ID NO:64); OXM327 (SEQ ID NO:66); or OXM329 (SEQ ID NO:67). 
     
     
         27 . The peptide analog of  claim 19 , wherein the peptide analog is OXM345 (SEQ ID NO:69); OXM355 (SEQ ID NO:70); OXM357 (SEQ ID NO:71); OXM359 (SEQ ID NO:72); OXM361 (SEQ ID NO:73); OXM373 (SEQ ID NO:74); OXM374 (SEQ ID NO:75); OXM380 (SEQ ID NO:76); OXM381 (SEQ ID NO:77); OXM383 (SEQ ID NO:78); OXM388 (SEQ ID NO:79); OXM392 (SEQ ID NO:80); OXM395 (SEQ ID NO:81); OXM398 (SEQ ID NO:82); OXM399 (SEQ ID NO:83); OXM400 (SEQ ID NO:84); OXM401 (SEQ ID NO:85); OXM404 (SEQ ID NO:86); OXM406 (SEQ ID NO:87); OXM407 (SEQ ID NO:88); OXM408 (SEQ ID NO:89); OXM410 (SEQ ID NO:91); OXM411 (SEQ ID NO:92); OXM412 (SEQ ID NO:93); OXM414 (SEQ ID NO:95); OXM415 (SEQ ID NO:96); OXM416 (SEQ ID NO:97; OXM417 (SEQ ID NO:98); OXM418 (SEQ ID NO:99; OXM419 (SEQ ID NO:100; OXM420 (SEQ ID NO:101); or OXM421 (SEQ ID NO:102).