Gip-based mixed agonists for treatment of metabolic disorders and obesity

Abstract

Glucagon peptides that exhibit GIP agonist activity in addition to glucagon and/or GLP-1 activity are provided. Pharmaceutical compositions comprising such glucagon peptides and therapeutic methods of using such peptides are also provided.

1 - 147 . (canceled) 
     
     
         148 . An analog of glucagon (SEQ ID NO: 1) having GIP agonist activity comprising an acyl group, wherein the acyl group is attached to a spacer, wherein:
 (i) the spacer is attached to the side chain of the amino acid at position 10 of the analog; or   (ii) the analog comprises an extension of 1 to 21 amino acids C-terminal to the amino acid at position 29 and the spacer is attached to the side chain of an amino acid corresponding to one of positions 37-43 relative to SEQ ID NO: 1;   wherein the analog exhibits at least 1% activity of native GIP at the GIP receptor.   
     
     
         149 . An analog of glucagon (SEQ ID NO: 1) having GIP agonist activity, with the following modifications:
 (I)   (a) an amino acid modification at position 1 that confers GIP agonist activity,   (b) a lactam bridge between the side chains of amino acids at positions i and i+4 or between the side chains of amino acids at positions j and j+3, wherein i is 12, 13, 16, 17, 20 or 24, and wherein j is 17,   (c) amino acid modifications at one, two or all of positions 27, 28 and 29, and   (d) 1-9 further amino acid modifications,   wherein the EC50 of the analog for GIP receptor activation is about 10 nM or less, and wherein the EC50 of the analog at the GIP receptor is less than about 50-fold different from its EC50 at the GLP-1 receptor; or   (II)   (a) an amino acid modification at position 1 that confers GIP agonist activity,   (b) one, two, three, or all of the amino acids at positions 16, 20, 21, and 24 of the analog is substituted with an α,α-disubstituted amino acid,   (c) amino acid modifications at one, two or all of positions 27, 28 and 29, and   (d) 1-9 further amino acid modifications,   wherein the analog is covalently linked to a hydrophilic moiety, wherein the EC50 of the analog for GIP receptor activation is about 10 nM or less, and wherein the EC50 of the analog at the GIP receptor is less than about 50-fold different from its EC50 at the GLP-1 receptor; or   (III)   (a) an amino acid modification at position 1 that confers GIP agonist activity,   (b) an amino acid substitution of Ser at position 16 with an amino acid of Formula IV:   
       
         
           
           
               
               
           
         
         wherein n is 1 to 7, wherein each of R1 and R2 is independently selected from the group consisting of H, C 1 -C 18  alkyl, (C 1 -C 18  alkyl)OH, (C 1 -C 18  alkyl)NH 2 , (C 1 -C 18  alkyl)SH, (C 0 -C 4  alkyl)(C 3 -C 6 )cycloalkyl, (C 0 -C 4  alkyl)(C 2 -C 5  heterocyclic), (C 0 -C 4  alkyl)(C 6 -C 10  aryl)R 7 , and (C 1 -C 4  alkyl)(C 3 -C 9  heteroaryl), wherein R 7  is H or OH, and the side chain of the amino acid of Formula IV comprises a free amino group, 
         (c) an amino acid substitution of the Gln at position 20 with an alpha, alpha-disubstituted amino acid, 
         (d) amino acid modifications at one, two or all of positions 27, 28 and 29, and 
         (e) 1-9 further amino acid modifications, 
         wherein the analog is covalently linked to a hydrophilic moiety, wherein the EC50 of the analog for GIP receptor activation is about 10 nM or less, and wherein the EC50 of the analog at the GIP receptor is less than about 50-fold different from its EC50 at the GLP-1 receptor; or 
         (IV) 
         (a) an amino acid modification at position 1 that confers GIP agonist activity, 
         (b) an amino acid substitution of Ser at position 16 with an amino acid of Formula IV: 
       
       
         
           
           
               
               
           
         
         wherein n is 1 to 7, wherein each of R1 and R2 is independently selected from the group consisting of H, C 1 -C 18  alkyl, (C 1 -C 18  alkyl)OH, (C 1 -C 18  alkyl)NH 2 , (C 1 -C 18  alkyl)SH, (C 0 -C 4  alkyl)(C 3 -C 6 )cycloalkyl, (C 0 -C 4  alkyl)(C 2 -C 5  heterocyclic), (C 0 -C 4  alkyl)(C 6 -C 10  aryl)R 7 , and (C 1 -C 4  alkyl)(C 3 -C 9  heteroaryl), wherein R 7  is H or OH, and the side chain of the amino acid of Formula IV comprises a free amino group, 
         (c) an amino acid substitution of the Gln at position 20 with an alpha, alpha-disubstituted amino acid, 
         (d) amino acid modifications at one, two or all of positions 27, 28 and 29, and 
         (e) 1-9 further amino acid modifications, 
         wherein said glucagon analog is not covalently linked to a hydrophilic moiety, and the EC50 of the analog for GIP receptor activation is about 10 nM or less, and wherein the EC50 of the analog at the GIP receptor is less than about 50-fold different from its EC50 at the GLP-I receptor. 
       
     
     
         150 . The analog of  claim 149 , wherein (i) the amino acid of Formula IV in (b) is homoLys, Lys, Orn, or 2,4-diaminobutyric acid (Dab); (ii) the alpha, alpha disubstituted amino acid is AIB, or (iii) a combination thereof. 
     
     
         151 . An analog comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 99-141, 144-164, 166, 192-207, 209-221 and 223, or from the group consisting of SEQ ID NOs: 167-169, 173-178 and 225, or the group consisting of SEQ ID NOs: 5-94. 
     
     
         152 . The analog of  claim 149 , wherein the hydrophilic moiety is a polyethylene glycol (PEG). 
     
     
         153 . The analog of  claim 152 , wherein the PEG has a molecular weight of about 20,000 Daltons to about 40,000 Daltons. 
     
     
         154 . The analog of  claim 149 , wherein the GIP potency of the analog is within about 15-fold of GLP-1 potency of the analog. 
     
     
         155 . A conjugate, a dimer or a fusion peptide comprising an analog of  claim 149 , or a combination thereof. 
     
     
         156 . A pharmaceutical composition comprising the analog of  claim 149 , or a dimer, a conjugate, or a fusion peptide comprising the analog, or a combination thereof, and a pharmaceutically acceptable carrier. 
     
     
         157 . A kit comprising a pharmaceutical composition of  claim 149  and a device for administering said pharmaceutical composition to a patient. 
     
     
         158 . A method of reducing weight gain or inducing weight loss, comprising administering to a patient in need thereof a pharmaceutical composition of  claim 149  in an amount effective to reduce weight gain or induce weight loss. 
     
     
         159 . A method of treating diabetes, comprising administering to a patient in need thereof a pharmaceutical composition of  claim 149  in an amount effective to lower blood glucose levels. 
     
     
         160 . The analog of  claim 149 , wherein
 (a) the amino acid at position 1 is an amino acid lacking an imidazole side chain or a large, aromatic amino acid,   (b) the α,α-disubstituted amino acid is AIB,   (c) the Met at position 27 is substituted with a large aliphatic amino acid   (d) the Asn at position 28 is substituted with a small aliphatic amino acid, and   (e) the Thr at position 29 is substituted with a small aliphatic amino acid,   (f) or a combination thereof.   
     
     
         161 . The analog of  claim 149 , wherein the amino acid at position 1 is Tyr, the amino acid at position 27 is Leu, the amino acid at position 28 is Ala, the amino acid at position 29 is Gly, or a combination thereof. 
     
     
         162 . The analog of  claim 149 , wherein (i) the amino acid of Formula IV in (b) is homoLys, Lys, Orn, or 2,4-diaminobutyric acid (Dab); (ii) the alpha, alpha disubstituted amino acid is AIB, or (iii) a combination thereof. 
     
     
         163 . The analog of  claim 149 , further comprising the amino acid sequence of GPSSGAPPPS (SEQ ID NO: 95) or XGPSSGAPPPS (SEQ ID NO: 96) C-terminal to the amino acid at position 29. 
     
     
         164 . The glucagon analog of  claim 149 , comprising the amino acid sequence according to any one of SEQ ID NOS: 227, 228, 229 or 230 and an extension of 1 to 21 amino acids C-terminal to the amino acid at position 29, further comprising up to 6 further amino acid modifications, wherein the EC50 of the analog for GIP receptor activation is about 10 nM or less. 
     
     
         165 . The analog of  claim 149 , comprising one or more of the following modifications:
 (a) Ser at position 2 substituted with D-Ser, Ala, D-Ala, Gly, N-methyl-Ser, AIB, Val, or α-amino-N-butyric acid;   (b) Gln at position 3 substituted with Glu;   (c) substitution of the amino acid Tyr at position 10 with an amino acid comprising a side chain covalently linked to an acyl group or alkyl group;   (d) addition of an amino acid comprising a side chain covalently linked to an acyl group or alkyl group as the C-terminal amino acid of the analog;   (e) Lys at position 12 substituted with Ile;   (f) Arg at position 17 substituted with Gln;   (g) Arg at position 18 substituted with Ala;   (h) Asp at position 21 substituted with Glu;   (i) Gln at position 24 substituted with Asn; and   (j) replacement of the carboxylic acid of the C-terminal amino acid with a charge-neutral group.   
     
     
         166 . The analog of  claim 163 , comprising (a) an amino acid modification at position 2 that confers resistance to DPP-IV, and (b) either (i) an amino acid covalently linked to an acyl group or alkyl group or (ii) a hydrophilic moiety linked to the amino acid at position 24.