Superagonist polypeptide analogs of adrenomedullin and intermedin peptide hormones

Abstract

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

1 . A CLR/RAMP receptor agonist peptide, comprising a structure of Formula I:
   B a -C a -D a   (I)
   wherein:   B a  is a modified N-terminal fragment of adrenomedullin 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; and   D a  is a modified C-terminal fragment of intermedin (IMD) comprising from 3-6 amino acid residues with a C-terminal amide, where at least one amino acid of the C-terminal fragment is histidine (His), proline (P), serine (Ser), tyrosine (Tyr).   
     
     
         2 . (canceled) 
     
     
         3 . 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, or an amino string of KTKKTLRT; B 1  is selected from the group consisting of an empty residue, histidine (His), acylated histidine (acy-His), double acylated histidine (acy-His(acy)), ace-histidine(acy) (ace-His(acy)), mini-PEG-acylated-histidine (mini-PEG-His(acy)), arginine (Arg), acylated arginine (acy-Arg), double acylated arginine (acy-Arg(acy)), ace-arginine(acy) (ace-Arg(acy)), mini-PEG-acylated-arginine (mini-PEG-Arg(acy)), lysine (Lys), acylated lysine (acy-Lys), double acylated lysine (acy-Lys(acy)), ace-lysine(acy) (ace-Lys(acy)), and mini-PEG-acylated-lysine (mini-PEG-Lys(acy)); B 2  is selected from the group consisting of glycine (Gly) and an empty residue; B 4  is selected from the group consisting of arginine (Arg), histidine (His), and lysine (Lys); B 5  is selected from the group consisting of phenylalanine (Phe) and leucine (Leu); B 7  is selected from the group consisting of serine (Ser), threonine (Thr) and tyrosine (Tyr); B 9  is selected from the group consisting of serine (Ser), threonine (Thr) and tyrosine (Tyr), glutamine (Gln), and asparagine (Asn); B 10  is selected from the group consisting of valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), and leucine (Leu); B 11  is selected from the group consisting of glutamine (Gln), and asparagine (Asn); B 12  is selected from the group consisting of histidine (His), arginine (Arg), lysine (Lys), glutamine (Gln), and asparagine (Asn); B 13  is selected from the group consisting of valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), and leucine (Leu); B 14  is selected from the group consisting of valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), leucine (Leu), serine (Ser), threonine (Thr), and tyrosine (Tyr); B 15  is selected from the group consisting of histidine (His), arginine (Arg), and lysine (Lys); B 16  is selected from the group consisting of an empty residue, glutamine (Gln), and asparagine (Asn); B 17  is selected from the group consisting of valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), and leucine (Leu); B 18  is selected from the group consisting of tryptophan (Trp), phenylalanine (Phe), serine (Ser), threonine (Thr), and tyrosine (Tyr); B 19  is selected from the group consisting of glutamine (Gln), glutamic acid (Glu), aspartic acid (Asp), and asparagine (Asn); B 20  is selected from the group consisting of tryptophan (Trp), phenylalanine (Phe), valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), and leucine (Leu); B 21  is selected from the group consisting of serine (Ser), threonine (Thr), and tyrosine (Tyr); methionine (Met), tryptophan (Trp), and phenylalanine (Phe). 
     
     
         4 . The agonist of  claim 1 , wherein the B a  sequence is selected from the group consisting of SEQ ID NO:2-16. 
     
     
         5 . The agonist of  claim 1  wherein the central core C a  comprises a fragment of human adrenomedullin or intermedin. 
     
     
         6 . (canceled) 
     
     
         7 . 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), and proline (Pro); 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 (lie), and leucine (Leu); C 3  is selected from the group consisting of an empty residue, aspartic acid (Asp), glutamic acid (Glu), glutamine (Gln), asparagine (Asn), valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), and leucine (Leu); C 4  is selected from the group consisting of an empty residue, histidine (His), arginine (Arg), and lysine (Lys); C 5  is selected from the group consisting of an empty residue, aspartic acid (Asp), glutamic acid (Glu), glutamine (Gln), and asparagine (Asn); C 6  is selected from the group consisting of an empty residue, aspartic acid (Asp), glutamic acid (Glu), glutamine (Gln), and asparagine (Asn); C 7  is selected from the group consisting of an empty residue, valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), leucine (Leu), serine (Ser), threonine (Thr), and tyrosine (Tyr); C 8  is selected from the group consisting of an empty residue, valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), and leucine (Leu); C 9  is selected from the group consisting of an empty residue, proline (Pro), valine (VI), alanine (Ala), glycine (Gly), isoleucine (lie), and leucine (Leu); C 10  is selected from the group consisting of an empty residue, valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), leucine (Leu), histidine (His), arginine (Arg), and lysine (Lys); 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), and tyrosine (Tyr); C 12  is selected from the group consisting of an empty residue, histidine (His), arginine (Arg), lysine (Lys), and proline (Pro). 
     
     
         8 . (canceled) 
     
     
         9 . The agonist of  claim 7  wherein the sequence of C a  has at least 60% sequence identity a sequence selected from SEQ ID NO:18-26. 
     
     
         10 . The agonist of  claim 1  wherein D a  comprises: D 1 -D 2 -D 3 -D 4 -D 5 -D 6 -NH 2  (SEQ ID NO: 27) where: D 1  is selected from the group consisting of an empty residue, Ser, Thr, and Tyr; D 2  is selected from the group consisting of an empty residue, Ser, Thr, and Tyr; D 3  is selected from the group consisting of an empty residue, Pro, Val, Ala, Gly, lie, and Leu; D 4  is selected from the group consisting of an empty residue, His, Arg, and Lys; D 5  is selected from the group consisting of an empty residue, Ser, Thr, and Tyr; and D 6  is selected from the group consisting of Ser, Thr, and Tyr. 
     
     
         11 - 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, and 110. 
     
     
         18 . A CLR/RAMP receptor superantagonist comprising a structure of Formula II: B b -C 1 -D b  (II), wherein B b  is an N-terminal fragment of adrenomedullin peptide family member comprising from four to thirteen amino acid residues; C b  is a central core consist of 3-12 amino acids; and D b  is a modified C-terminal fragment of intermedin (IMD) comprising 3-6 amino acid residues with a C-terminal amide, where at least one amino acid of the C-terminal fragment is histidine (His), proline (P), serine (Ser), tyrosine (Tyr). 
     
     
         19 . The antagonist of  claim 18  wherein the N-terminal fragment B b  comprises: B 0 -B 1 -B 2 -B 3 -B 4 -B 5 -B 6 -B 7 -B 8 -B 9 -B 10 -B 11 -B 12  where: B 0  is selected from the group consisting of an empty residue, acylated histidine (acy-His), acylated arginine (acy-Arg), acylated lysine (acy-Lys), acylated serine (acy-Ser), acylated threonine (acy-Thr), acylated tyrosine (acy-Tyr), acylated aspartic acid (acy-Asp), acylated glutamic acid (acy-Glu), acylated glutamine (acy-Gln), acylated asparagine (acy-Asn); acylated valine (acy-Val), acylated alanine (acy-Ala), acylated glycine (acy-Gly), acylated isoleucine (acy-Ile), acylated leucine (acy-Leu), acylated phenylalanine (acy-Phe), acylated tryptophan (acy-Trp), acylated proline (acy-Pro), acylated methionine (acy-Met), acylated cysteine (acy-Cys), double acylated histidine (acy-His(acy)), ace-histidine(acy) (ace-His(acy)), mini-PEG-acylated-histidine (mini-PEG-His(acy)), double acylated arginine (acy-Arg(acy)), ace-arginine(acy) (ace-Arg(acy)), mini-PEG-acylated-arginine (mini-PEG-Arg(acy)), lysine (Lys), double acylated lysine (acy-Lys(acy)), ace-lysine(acy) (ace-Lys(acy)), and mini-PEG-acylated-lysine (mini-PEG-Lys(acy); B 1  is selected from the group consisting of an empty residue, valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), and leucine (Leu); B 2  is selected from the group consisting of an empty residue, glutamine (Gln), glutamic acid (Glu), aspartic acid (Asp), and asparagine (Asn); B 3  is selected from the group consisting of an empty residue, histidine (His), arginine (Arg), lysine (Lys), glutamine (Gln), and asparagine (Asp); B 4  is selected from the group consisting of an empty residue, valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), and leucine (Leu); B 5  is selected from the group consisting of an empty residue, valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), leucine (Leu), serine (Ser), threonine (Thr), and tyrosine (Tyr); B 6  is selected from the group consisting of an empty residue, histidine (His), arginine (Arg), and lysine (Lys); B 7  is selected from the group consisting of an empty residue, glutamine (Gln), and asparagine (Asn); B 8  is selected from the group consisting of an empty residue, valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), and leucine (Leu); B 9  is selected from the group consisting of an empty residue, tryptophan (Trp), phenylalanine (Phe), serine (Ser), threonine (Thr), and tyrosine (Tyr); B 10  is selected from the group consisting of an empty residue, glutamine (Gln), glutamic acid (Glu), aspartic acid (Asp), and asparagine (Asn); B 11  is selected from the group consisting of an empty residue, tryptophan (Trp), phenylalanine (Phe), valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), and leucine (Leu); B 12  is selected from the group consisting of an empty residue, serine (Ser), threonine (Thr), and tyrosine (Tyr); methionine (Met), tryptophan (Trp), and phenylalanine (Phe). 
     
     
         20 . (canceled) 
     
     
         21 . The antagonist of  claim 18  wherein C b  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, glutamine (Gln), glutamic acid (Glu), aspartic acid (Asp), asparagine (Asn), and proline (Pro); 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 (lie), and leucine (Leu); C 3  is selected from the group consisting of an empty residue, glutamine (Gln), glutamic acid (Glu), aspartic acid (Asp), asparagine (Asn), valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), and leucine (Leu); C 4  is selected from the group consisting of an empty residue, histidine (His), arginine (Arg), and lysine (Lys); C 5  is selected from the group consisting of an empty residue, aspartic acid (Asp), glutamic acid (Glu), glutamine (Gln), and asparagine (Asn); C 6  is selected from the group consisting of an empty residue, aspartic acid (Asp), glutamic acid (Glu), glutamine (Gln), and asparagine (Asn); C 7  is selected from the group consisting of an empty residue, valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), leucine (Leu), serine (Ser), threonine (Thr), and tyrosine (Tyr); C is selected from the group consisting of an empty residue, valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), and leucine (Leu); C 9  is selected from the group consisting of an empty residue, proline (Pro), valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), and leucine (Leu); C 10  is selected from the group consisting of an empty residue, valine (Val), alanine (Ala), glycine (Gly), isoleucine (lie), leucine (Leu), histidine (His), arginine (Arg), and lysine (Lys); 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), and tyrosine (Tyr); C 12  is selected from the group consisting of an empty residue, histidine (His), arginine (Arg), lysine (Lys), and proline (Pro). 
     
     
         22 . (canceled) 
     
     
         23 . The antagonist of  claim 18  wherein C b  comprises a sequence with at least 60% sequence identity with DKDKDNSAPVDP (SEQ ID NO: 19). 
     
     
         24 . The antagonist of  claim 18  wherein D b  comprises: D 1 -D 2 -D 3 -D 4 -D 5 -D 6 -NH 2  (SEQ ID NO: 27) where: D 1  is selected from the group consisting of an empty residue, Ser, Thr, and Tyr; D 2  is selected from the group consisting of an empty residue, Ser, Thr, and Tyr; D 3  is selected from the group consisting of an empty residue, Pro, Val, Ala, Gly, lie, and Leu; D 4  is selected from the group consisting of an empty residue, His, Arg, and Lys; D 5  is selected from the group consisting of an empty residue, Ser, Thr, and Tyr; and D 6  is selected from the group consisting of Ser, Thr, and Tyr. 
     
     
         25 . (canceled) 
     
     
         26 . The antagonist of  claim 24  wherein D b  comprises a sequence with greater than 60% sequence identity with Ser-Ser-Pro-His-Ser-Tyr-NH 2 . 
     
     
         27 . A CLR/RAMP receptor superantagonist comprising a structure of Formula III: B 0 -B 1 -B 2 -B 3 -B 4 -B 5 -B 6 -B 7 -B 8 -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 -B 22 -B 23 -B 24 -B 25 -B 26 -B 27 -B 28 - NH 2  (III), where:
 B 0  is selected from the group consisting of an empty residue, acylated histidine (acy-His), acylated arginine (acy-Arg), acylated lysine (acy-Lys), acylated serine (acy-Ser), acylated threonine (acy-Thr), acylated tyrosine (acy-Tyr), acylated aspartic acid (acy-Asp), acylated glutamic acid (acy-Glu), acylated glutamine (acy-Gln), acylated asparagine (acy-Asn); acylated valine (acy-Val), acylated alanine (acy-Ala), acylated glycine (acy-Gly), acylated isoleucine (acy-Ile), acylated leucine (acy-Leu), acylated phenylalanine (acy-Phe), acylated tryptophan (acy-Trp), acylated proline (acy-Pro), acylated methionine (acy-Met), acylated cysteine (acy-Cys), double acylated histidine (acy-His(acy)), ace-histidine(acy) (ace-His(acy)), mini-PEG-acylated-histidine (mini-PEG-His(acy)), double acylated arginine (acy-Arg(acy)), ace-arginine(acy) (ace-Arg(acy)), mini-PEG-acylated-arginine (mini-PEG-Arg(acy)), lysine (Lys), double acylated lysine (acy-Lys(acy)), ace-lysine(acy) (ace-Lys(acy)), and mini-PEG-acylated-lysine (mini-PEG-Lys(acy); B 1  is selected from the group consisting of an empty residue, Val, Ala, Gly, lie, and Leu B 2  is selected from the group consisting of an empty residue, Gln, Glu, Asp, and Asn; B 3  is selected from the group consisting of an empty residue, His, Arg, Lys, Gln, and Asp; B 4  is selected from the group consisting of an empty residue, Val, Ala, Gly, lie, and Leu; B 5  is selected from the group consisting of an empty residue, Val, Ala, Gly, lie, Leu, Ser, Th, and Tyr; B 6  is selected from the group consisting of an empty residue, His, Arg, and Lys; B 7  is selected from the group consisting of an empty residue, Gln, and Asn, His, Arg, and Lys; B 8  is selected from the group consisting of an empty residue, Val, Ala, Gly, lie, and Leu; B 9  is selected from the group consisting of an empty residue, Trp, Phe, Ser, Thr, and Tyr; B 10  is selected from the group consisting of an empty residue, Gln, Glu, Asp, and Asn; B 11  is selected from the group consisting of an empty residue, Trp, Phe, Val, Ala, Gly, lie, and Leu; B 12  is selected from the group consisting of an empty residue, Ser, Thr, and Tyr; Met, Trp, and Phe; B 13  is selected from the group consisting of an empty residue, Gln, Glu, Asp, and Asn, Val, Ala, Gly, lie, and Leu; B 14  is selected from the group consisting of an empty residue, His, Arg, Lys, Val, Ala, Gly, lie, Leu, and Pro; B 15  is selected from the group consisting of an empty residue, Gln, Glu, Asp, Asn, Val, Ala, Gly, lie, and Leu; B 16  is selected from the group consisting of an empty residue, Val, Ala, Gly, lie, and Leu; B 17  is selected from the group consisting of an empty residue, Ser, Thr, and Tyr; B 18  is selected from the group consisting of an empty residue, Val, Ala, Gly, lie, and Leu; B 19  is selected from the group consisting of an empty residue, Val, Ala, Gly, lie, Leu, and Pro; B 20  is selected from the group consisting of an empty residue, His, Arg, Lys, Val, Ala, Gly, lie, and Leu; B 21  is selected from the group consisting of an empty residue, Ser, Thr, Tyr, Gln, Glu, Asp, and Asn; B 22  is selected from the group consisting of an empty residue, His, Arg, Lys, Val, Ala, Gly, lie, Leu, and Pro; B 23  is selected from the group consisting of an empty residue, Ser, Thr, Tyr, Val, Ala, Gly, lie, and Leu; B 24  is selected from the group consisting of an empty residue, Ser, Thr, and Tyr; B 25  is selected from the group consisting of an empty residue, Val, Ala, Gly, lie, Leu, and Pro; B 26  is selected from the group consisting of an empty residue, His, Arg, Lys, Gln, Glu, Asp, and Asn; B 27  is selected from the group consisting of an empty residue, Val, Ala, Gly, lie, Leu, Ser, Thr, and Tyr; B 28  is selected from the group consisting of an empty residue, Ser, Thr, and Tyr.   
     
     
         28 - 31 . (canceled) 
     
     
         32 . A CLR/RAMP receptor superagonist, comprising: a stereoisomer, derivative, or peptidomimetics of an amino acid sequence selected from SEQ ID NOS: 77-78, 112, 114, 119, 120-125, and 139. 
     
     
         33 - 37 . (canceled)