Peptide analogs

Abstract

Analogs for CLR/RAMP receptor ligands are provided that have agonist, superagonist, antagonist, super-antagonist, or multiple receptor modulatng activity. The analogs can be selective for one or more CLR/RAMP receptors, or can be pan-specific for multiple G protein-coupled receptors.

1 . A CLR/RAMP receptor agonist peptide, comprising a structure of Formula I:
   R1-B a -C a -D a -R2   (I)
   wherein:   B a  is a modified N-terminal fragment of adrenomedullin (ADM) peptide family member comprising from twenty to twenty-eight amino acid residues, wherein two amino acid residues of the fragment are cysteine (Cys), wherein the C-terminal residue of the fragment is threonine (Thr);   C a  is a central core consist of 3-12 amino acids;   D a  is a modified C-terminal fragment of intermedin (IMD) or ADM comprising from 3-6 amino acid residues with a C-terminal amide, where at least one amino acid of the C-terminal fragment is proline (P), serine (Ser), tyrosine (Tyr);   R1 is a functional group comprising a structure of Formula (W′)(X′)n(Y′)n′(Z′)n″, wherein W is a fatty acid, a long-chain fatty dicarboxylic acid, a fatty acid derivative or empty; X′ is a PEG group, glutamic acid, y-glutamic acid, a proteinogenic or non-proteinogenic amino acid, Lys, or empty; Y′ is a PEG group, glutamic acid, y-glutamic acid, a proteinogenic or non-proteinogenic amino acid, Lys, or empty; Z′ is a proteinogenic amino acid, a non-proteinogenic amino acid, Lys, or empty; and each of n, n′ and n″ is an independently selected integer from 1 to 20, where R1 can be located on any side chain group of an amino acid in the peptide; and   R2 is an NH 2  group.   
     
     
         2 . The agonist peptide of  claim 1 , wherein W has a polar group at the end of the chain distal from connection X′, said polar group comprising a carboxylic acid or a carboxylic acid bioisostere, a phosphonic acid, or a sulfonic acid group. 
     
     
         3 . The agonist peptide of  claim 1 , wherein W′ comprises a lipid selected from hexadecanoyl; 17-carboxy-heptadecanoyl; 15-carboxy-pentadecanoyl, Octadecanoyl, Eicosanoyl; and [19-carboxy-nonadecanoyl]. 
     
     
         4 . The agonist of  claim 1 , wherein the N-terminal fragment B a  comprises: B 0 -B 1 -B 2 -C-B 4 -B 5 -G-B 7 -C-B 9 -B 10 -B 11 -B 12 -B 13 -B 14 -B 15 -B 16 -B 17 -B 18 -B 19 -B 20 -B 21  (SEQ ID NO: 1) where: B 0  is absent or present (SEQ ID NO:236), Lys (SEQ ID NO:1), or an amino string of KTKKTLRT (SEQ ID NO:235); B 1  is selected from the group consisting of an empty residue, histidine (His), acylated histidine (acy-His), arginine (Arg), acylated arginine (acy-Arg), lysine (Lys), or acylated lysine (acy-Lys); B 2  is selected from the group consisting of glycine (Gly), a non-proteinogenic amino acid, and an empty residue; B 4  is selected from the group consisting of arginine (Arg), histidine (His), and lysine (Lys), a non-proteinogenic amino acid and an empty residue; B 5  is selected from the group consisting of phenylalanine (Phe), leucine (Leu), tyrosine (Tyr), a non-proteinogenic amino acid and an empty residue; B 7  is selected from the group consisting of serine (Ser), threonine (Thr), tyrosine (Tyr), a non-proteinogenic amino acid and an empty residue; B 9  is selected from the group consisting of serine (Ser), threonine (Thr), tyrosine (Tyr), glutamine (Gin), asparagine (Asn), a non-proteinogenic amino acid and an empty residue; B 10  is selected from the group consisting of valine (Val), alanine (Ala), glycine (Gly), isoleucine (Ile), leucine (Leu), a non-proteinogenic amino acid and an empty residue; B 11  is selected from the group consisting of glutamine (Gln), asparagine (Asn), Asp, Glu, a non-proteinogenic amino acid and an empty residue; B 12  is selected from the group consisting of histidine (His), arginine (Arg), lysine (Lys), glutamine (Gin), asparagine (Asn), a non-proteinogenic amino acid and an empty residue; B 13  is selected from the group consisting of valine (Val), alanine (Ala), glycine (Gly), isoleucine (Ile), leucine (Leu), a non-proteinogenic amino acid and an empty residue; B 14  is selected from the group consisting of valine (Val), alanine (Ala), glycine (Gly), isoleucine (Ile), leucine (Leu), serine (Ser), threonine (Thr), tyrosine (Tyr), a non-proteinogenic amino acid and an empty residue; B 15  is selected from the group consisting of histidine (His), arginine (Arg), lysine (Lys), a non-proteinogenic amino acid and an empty residue; B 16  is selected from the group consisting of an empty residue, glutamine (Gin), asparagine (Asn), Asp, Glu, a non-proteinogenic amino acid and an empty residue; B 17  is selected from the group consisting of valine (Val), alanine (Ala), glycine (Gly), isoleucine (Ile), leucine (Leu), a non-proteinogenic amino acid and an empty residue; B 18  is selected from the group consisting of tryptophan (Trp), phenylalanine (Phe), serine (Ser), threonine (Thr), tyrosine (Tyr), a non-proteinogenic amino acid and an empty residue; B 19  is selected from the group consisting of glutamine (Gin), glutamic acid (Glu), aspartic acid (Asp), asparagine (Asn), a non-proteinogenic amino acid and an empty residue; B 20  is selected from the group consisting of tryptophan (Trp), phenylalanine (Phe), valine (Val), alanine (Ala), glycine (Gly), isoleucine (Ile), leucine (Leu), a non-proteinogenic amino acid and an empty residue; B 21  is selected from the group consisting of serine (Ser), threonine (Thr), and tyrosine (Tyr); methionine (Met), tryptophan (Trp), phenylalanine (Phe), a non-proteinogenic amino acid and an empty residue. 
     
     
         5 . The agonist of  claim 1 , wherein the B a  sequence is selected from the group consisting of SEQ ID NOS:2-16. 
     
     
         6 . The agonist of  claim 1  wherein the central core C a  comprises a fragment of human adrenomedullin or intermedin. 
     
     
         7 . The agonist of  claim 5  wherein the fragment of C a  comprises 3 to 12 amino acids. 
     
     
         8 . The agonist of  claim 1  wherein C a  comprises: C 1 -C 2 -C 3 -C 4 -C 5 -C 6 -C 7 -C 8 -C 9 -C 10 -C 11 -C 12  (SEQ ID NO: 17) where: C 1  is selected from the group consisting of an empty residue, aspartic acid (Asp), glutamic acid (Glu), glutamine (Gln), asparagine (Asn), proline (Pro), and a non-proteinogenic amino acid; C 2  is selected from the group consisting of an empty residue, histidine (His), arginine (Arg), lysine (Lys), valine (Val), alanine (Ala), glycine (Gly), isoleucine (Ile), leucine (Leu) and a non-proteinogenic amino acid; C 3  is selected from the group consisting of an empty residue, aspartic acid (Asp), glutamic acid (Glu), glutamine (Gin), asparagine (Asn), valine (Val), alanine (Ala), glycine (Gly), isoleucine (Ile), leucine (Leu) and a non-proteinogenic amino acid; C 4  is selected from the group consisting of an empty residue, histidine (His), arginine (Arg), lysine (Lys) and a non-proteinogenic amino acid; C 5  is selected from the group consisting of an empty residue, aspartic acid (Asp), glutamic acid (Glu), glutamine (Gin), asparagine (Asn) and a non-proteinogenic amino acid; C 6  is selected from the group consisting of an empty residue, aspartic acid (Asp), glutamic acid (Glu), glutamine (Gin), asparagine (Asn) and a non-proteinogenic amino acid; C 7  is selected from the group consisting of an empty residue, valine (Val), alanine (Ala), glycine (Gly), isoleucine (Ile), leucine (Leu), serine (Ser), threonine (Thr), tyrosine (Tyr) and a non-proteinogenic amino acid; C 8  is selected from the group consisting of an empty residue, valine (Val), alanine (Ala), glycine (Gly), isoleucine (Ile), leucine (Leu) and a non-proteinogenic amino acid; C 9  is selected from the group consisting of an empty residue, proline (Pro), valine (VI), alanine (Ala), glycine (Gly), isoleucine (Ile), leucine (Leu) and a non-proteinogenic amino acid; C 10  is selected from the group consisting of an empty residue, valine (Val), alanine (Ala), glycine (Gly), isoleucine (Ile), leucine (Leu), histidine (His), arginine (Arg), lysine (Lys) and a non-proteinogenic amino acid; C 11  is selected from the group consisting of an empty residue, aspartic acid (Asp), glutamic acid (Glu), glutamine (Gln), asparagine (Asn), serine (Ser), threonine (Thr), tyrosine (Tyr) and a non-proteinogenic amino acid; C 12  is selected from the group consisting of an empty residue, histidine (His), arginine (Arg), lysine (Lys), proline (Pro) and a non-proteinogenic amino acid. 
     
     
         9 . The agonist of  claim 8  wherein the sequence of Ca is selected from SEQ ID NOS:18-26. 
     
     
         10 . The agonist of  claim 7  wherein the sequence of C a  has at least 60% sequence identity with a sequence selected from SEQ ID NOS:18-26. 
     
     
         11 . The agonist of  claim 1  wherein D a  comprises: D 1 -D 2 -D 3 -D 4 -D 5 -D 6 (SEQ ID NO: 27) where: D 1  is selected from the group consisting of an empty residue, Val, Ala, Gly, Ile, Leu, Ser, Thr, Tyr and a non-proteinogenic amino acid; D 2  is selected from the group consisting of an empty residue, Ser, Thr, Tyr and a non-proteinogenic amino acid; D 3  is selected from the group consisting of an empty residue, Pro, Val, Ala, Gly, Ile, Leu and a non-proteinogenic amino acid; D 4  is selected from the group consisting of an empty residue, Asn, Gln, His, Arg, Lys and a non-proteinogenic amino acid; D 5  is selected from the group consisting of an empty residue, Val, Ala, Gly, Ile, Leu, Ser, Thr, Tyr and a non-proteinogenic amino acid; and D 6  is selected from the group consisting of Ser, Thr, Tyr and a non-proteinogenic amino acid. 
     
     
         12 . The agonist of  claim 11  wherein D 1 -D 2 -D 3 -D 4 -D 5 -D 6  (SEQ ID NO: 27) is Ser-Ser-Pro-His-Ser-Tyr (SEQ ID NO:237) or Ile-Ser-Pro-Gln-Gly-Tyr SEQ ID NO:238 . 
     
     
         13 . The agonist of  claim 1 , comprising a first peptide fragment having 21 amino acid residues or less from adrenomedullin, a second peptide fragment having 3-12 amino acids from adrenomedullin or intermedin, and a third peptide fragment having six amino acid residues or less from intermedin or adrenomedullin. 
     
     
         14 . The agonist of  claim 1  comprising a sequence selected from SEQ ID NOS: 28-51, 92, 147, 150-159, and 213 or a pharmaceutically acceptable salt thereof. 
     
     
         15 . A modified CLR/RAMP2 receptor-selective superagonist, said agonist comprising a sequence selected from SEQ ID NOS: 69, 70, 94, 101, 103, 110, 210, 211, and 214. 
     
     
         16 . (canceled) 
     
     
         17 . A CLR/RAMP receptor superagonist, comprising a stereoisomer, derivative, or peptidomimetic of an amino acid sequence selected from SEQ ID NOS: 28-51, 69-70, 92, 94, 101, 103, 110, 147, 150-159, 210, 211, 213, and 214. 
     
     
         18 - 75 . (canceled) 
     
     
         76 . The agonist of  claim 9  wherein D a  has greater than 60% sequence identity with Ser-Ser-Pro-His-Ser-Tyr (SEQ ID NO:237) or Ile-Ser-Pro-Gln-Gly-Tyr (SEQ ID NO:238).