US 5,844,074 A · Cyclic CRF agonists

Abstract

Novel cyclic CRF agonist peptides have the amino acid sequence: (cyclo 30-33)Ac-Pro-Pro-Ile-Ser-Leu-Asp-Leu-Thr-D-Phe-His-Leu-Leu-Arg-Glu-Val-Leu -Glu-Nle-Ala-Arg-Ala-Glu-Gln-Leu-Ala-Gln-R.sub.30 -Ala-R.sub.32 -R.sub.33 -Asn-Arg-Lys-Leu-Nle-Glu-Ile-Ile-NH.sub.2 wherein R.sub.30 is Glu or Cys; R.sub.32 is His or a D-amino acid such as D-His, D-Arg or similar; R.sub.33 is Lys, Orn or Cys. The N-terminus may be extended by Ser-Glu-Glu. Lys may be substituted for Arg.sup.23, and its side chain connected by a lactam bridge to Glu.sup.20 to form a bicyclic peptide. Certain disclosed CRF agonists include: (cyclo 30-33)Ac-Pro.sup.4, D-Phe.sup.12, Nle.sup.21,38, D-His.sup.32, Glu.sup.30, Lys.sup.33 !r/hCRF(4-41), (cyclo 30-33)Ac-Pro.sup.4, D-Phe.sup.12, Nle.sup.21,38, D-His.sup.32, Glu.sup.30, Orn.sup.33 !r/hCRF(4-41), (cyclo 30-33)Ac-Pro.sup.4, D-Phe.sup.12, Nle.sup.21,38, Cys.sup.30,33, D-His.sup.32 !r/hCRF(4-41), (bicyclo 20-23,30-33)Ac-Pro.sup.4, D-Phe.sup.12, Nle.sup.21,38, Lys.sup.23,33, Glu.sup.30, D-His.sup.32 !-r/hCRF(4-41), (cyclo 30-33)Ac-Pro.sup.4, D-Phe.sup.12, Nle.sup.21,38, CML.sup.27, Glu.sup.30, imBzlD-His.sup.32, Lys.sup.33 !r/hCRF(4-41) and (cyclo 30-33)Ac-Pro.sup.4, D-Phe.sup.12, Nle.sup.21,38, Glu.sup.30, D-Arg.sup.32, Lys.sup.33 !r/hCRF(4-41). Labelled agonists such as (cyclo 30-33)I.sup.125 Tyr.sup.o, D-Phe.sup.12, Nle.sup.21,38, Glu.sup.30, D-His.sup.32, Lys.sup.33 !r/hCRF and (cyclo 30-33)I.sup.125 D-Tyr.sup.3, D-Phe.sup.12, Nle.sup.21,38, Glu.sup.30, D-His.sup.32, Lys.sup.33 !r/hCRF(3-41) are useful in screening for more potent agonists.

Claims20

Dependent claims

A cyclic CRF agonist peptide which binds to CRF receptors CRF-RA and CRF-RB with an affinity greater than that of r/hCRF, which peptide has the formula A-Xaa.sub.c -Xaa-D-Xaa-Xaa.sub.c -B-NH.sub.2 wherein A is a sequence of 26 to 29 amino acid residues of the corresponding sequence of a peptide of the CRF family selected from the group consisting of residues 1-29, 2-29, 3-29 or 4-29 of mammalian and fish CRFs and fish urotensins and residues 1-28, 2-28 or 3-28 of sauvagine;

Xaa.sub.c are amino acid residues the side chains of which are linked in a cyclizing bond;
Xaa is a natural .alpha.-amino acid residue other than Cys;
D-Xaa is a residue of a D-isomer amino acid from the group consisting of D-isomers of natural .alpha.-amino acids other than Cys and unnatural aromatic .alpha.-amino acids, and B is a sequence of the last 8 amino acid residues of the C-terminal portion of one said peptide of the CRF family, wherein A may contain D-Phe or D-Leu instead of Phe or Leu at the position spaced from D-Xaa by 19 residues and may contain D-Pro instead of Pro at the position spaced from D-Xaa by 26 or 27 residues, wherein Nle may be substituted for Met in said selected peptide sequences, and wherein the N-terminus may be acylated.

A CRF agonist peptide having the amino acid sequence:

##STR2## wherein Y.sub.1 is H or an acylating agent having up to 15 carbon atoms;
Y.sub.2 is Glu, Asp, Gly, Glu-Glu, Asn-Asp, Gln-Glu, pGlu-Gly, Ser-Glu-Glu, Asn-Asp-Asp, Ser-Gln-Glu, or des-Y.sub.2 ;
R.sub.6 is Ile, Met or Nle;
R.sub.8 is Leu or Ile;
R.sub.11 is Thr or Ser;
R.sub.13 is His, Tyr or Glu;
R.sub.14 is CML or Leu;
R.sub.15 is CML or Leu;
R.sub.17 is Glu, CML, Asn or Lys;
R.sub.18 is Val, Nle or Met;
R.sub.19 is CML, Leu, Ile, Ala or Aib;
R.sub.20 is Glu, D-Glu, Cys or His;
R.sub.22 is Ala, D-Ala, Aib, Thr, Asp or Glu;
R.sub.23 is Arg, Cys, Orn or Lys;
R.sub.24 is Ala, Gln, Ile, Asn or Aib;
R.sub.25 is Asp or Glu;
R.sub.26 is Gln, Asn or Lys;
R.sub.27 is CML, Glu, Gln or Leu;
R.sub.28 is Ala, Lys, Arg or Aib;
R.sub.29 is Gln, Aib or Glu;
R.sub.30 is Glu;
R.sub.31 is Ala or Aib;
R.sub.32 is His, D-His, D-Arg, D-Amp, D-Iamp, D-2Nal, D-Glu, D-Ala or an L- or D-isomer of a natural .alpha.-amino acid other than Cys;
R.sub.33 is Lys or Orn;
R.sub.34 is Asn or Aib;
R.sub.36 is Lys, Orn, Arg, Har or Leu;
R.sub.37 is CML, Leu or Tyr;
R.sub.39 is Glu, Aib or Asp;
R.sub.40 is Ile, Aib, Thr, Glu, Ala, Val, Leu, Nle, Phe, Nva, Gly or Gln;
and R.sub.41 is Ala, Ile, Gly, Val, Leu, Nle, Phe, Nva or Gln;
wherein Tyr or D-Tyr may be optionally included at the N-terminus;
wherein D-Phe may be substituted by Phe, Leu, Tyr, D-Leu, D-Tyr, D-Cpa, D-Nal, D-Pal or another D-isomer .alpha.-amino acid, and wherein either Pro.sup.4 or Pros may be substituted by D-Pro;
provided that cyclizing bond may exist between R.sub.20 and R.sub.23.

30 more elements

A CRF agonist peptide having the amino acid sequence, or a nontoxic salt thereof:

(cyclo 30-33)Y.sub.1 -Y.sub.2 -Pro-Pro-R.sub.6 -Ser-R.sub.8 -Asp-Leu-R.sub.11 -D-Phe-R.sub.13 -R.sub.14 -R.sub.15 -Arg-R.sub.17 -R.sub.18 -R.sub.19 -R.sub.20 -R.sub.21 -R.sub.22 -R.sub.23 -R.sub.24 -R.sub.25 -R.sub.26 -R.sub.27 -R.sub.28 -R.sub.29 -R.sub.30 -R.sub.31 -R.sub.32 -R.sub.33 -R.sub.34 -Arg-R.sub.36 -R.sub.37 -Nle-R.sub.39 -R.sub.40 -R.sub.41 -NH.sub.2 wherein Y.sub.1 is Ac or hydrogen;
Y.sub.2 is Glu, Asp, Gly, Glu-Glu, Asn-Asp, Gln-Glu, pGlu-Gly, Ser-Glu-Glu, Asn-Asp-Asp, Ser-Gln-Glu, or des-Y.sub.2 ;
R.sub.6 is Ile, Met or Nle;
R.sub.8 is Leu or Ile;
R.sub.11 is Thr or Ser;
R.sub.13 is His, Tyr or Glu;
R.sub.14 is Leu or CML;
R.sub.15 is Leu or CML;
R.sub.17 is Glu, Lys, Asn or CML;
R.sub.18 is Val, Nle or Met;
R.sub.19 is Leu, Ile or CML;
R.sub.20 is His, D-Glu, Cys or Glu;
R.sub.21 is Nle, Met or Ile;
R.sub.22 is Ala, D-Ala, Aib, Thr, Asp or Glu;
R.sub.23 is Arg, Cys, Orn or Lys;
R.sub.24 is Ala, Asn, Gln, Ile or Aib;
R.sub.25 is Asp or Glu;
R.sub.26 is Gln, Asn or Lys;
R.sub.27 is Leu, Glu, Gln or CML;
R.sub.28 is Ala, Arg, Lys or Aib;
R.sub.29 is Gln, Aib or Glu;
R.sub.30 is Glu;
R.sub.31 is Ala or Aib;
R.sub.32 is His, D-His, Aib, D-Arg, D-2Nal, D-3Pal, D-Amp, D-Iamp, Gly, Tyr, D-Tyr, Ala, D-Ala or another aromatic D-isomer .alpha.-amino acid;
R.sub.33 is Lys or Orn;
R.sub.34 is Asn or Aib;
R.sub.36 is Lys, Orn, Arg, Har or Leu;
R.sub.37 is CML, Leu or Tyr;
R.sub.39 is Glu, Aib or Asp;
R.sub.40 is Ile, Aib, Thr, Glu, Ala, Val, Leu, Nle, Phe, Nva, Gly or Gln;
and R.sub.41 is Ala, Ile, Gly, Val, Leu, Nle, Phe, Nva or Gln;
wherein D-Leu or Phe or Leu may be substituted for D-Phe;
wherein D-Pro may be substituted for Pro.sup.4 or Pro.sup.5 ;
and wherein Tyr or D-Tyr may be optionally included at the N-terminus;
provided that a second cyclizing bond may exist between R.sub.20 and R.sub.23.

32 more elements

Dependencies are inferred from the claim text.

Cyclic CRF agonists

Life of this patent

Cyclic CRF agonists

Life of this patent

Abstract

Claims20