US 5,824,771 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, D-His or an equivalent .alpha.-amino acid; R.sub.33 is Lys, Orn or Cys. The N-terminus may be extended by Ser-Glu-Glu or shortened by up to three more residues. 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, Glu.sup.30, D-His.sup.32, Lys.sup.33 !r/hCRF(4-41), (cyclo 30-33)Ac-Ser.sup.7, D-Phe.sup.12, Nle.sup.21,38, Glu.sup.30, Lys.sup.33 !r/hCRF(7-41), (cyclo 30-33)Ac-Ser.sup.7, D-Phe.sup.12, Nle.sup.21,38, Glu.sup.30, D-His.sup.32, Lys.sup.33 !r/hCRF(7-41), (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-Ser.sup.7, D-Phe.sup.12, Nle.sup.18,21, Glu.sup.30, D-Ala.sup.32, Lys.sup.33 !.alpha.-helical CRF(7-41), and (cyclo 30-33)Ac-Ser.sup.7, D-Phe.sup.12, Nle.sup.21,38, CML.sup.27,40, Glu.sup.30, Lys.sup.33 !r/hCRF(7-41). Labelled agonists such as (cyclo 30-33)I.sup.125 Tyr.sup.0, 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 CRF agonists.

Claims25

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 Y.sub.1 -A-D-Xaa-B-Xaa.sub.c -Xaa.sub.a -Xaa.sub.b -Xaa.sub.c -C-NH.sub.2 wherein Y.sub.1 is an acyl group having not more than 15 carbon atoms;

A is Ser-Leu-Asp-Leu-Thr or Ser-Ile-Asp-Leu-Ser or Ser-Ile-Asp-Leu-Thr;
D-Xaa is D-Phe, D-2Nal or D-Leu;
B is a sequence of 17 amino acid residues that is found between Phe in the 12-position and Gln in position-30 of r/hCRF or the corresponding sequence of another peptide of the CRF family selected from the group consisting of residues 13-29 of other mammalian and fish CRFs and fish urotensins and residues 12-28 of sauvagine;
Xaa.sub.c represent a pair of amino acid residues, the side chains of which are linked in a cyclizing bond;
Xaa.sub.a is a natural .alpha.-amino acid residue other than Cys;
Xaa.sub.b is a residue of either (a) 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, or (b) a natural L-isomer .alpha.-amino acid;
and C is a sequence of the last 8 amino acid residues of the C-terminal portion of said peptide of the CRF family;
provided that Nle or Leu may be substituted for Met in B and in C.

5 more elements

A CRF agonist peptide having the formula:

(cyclo 30-33)Y.sub.1 -R.sub.6 -R.sub.7 -R.sub.8 -Asp-R.sub.10 -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 -R.sub.38 -R.sub.39 -R.sub.40 -R.sub.41 -NH.sub.2 wherein Y.sub.1 is an acyl group having not more than 15 carbon atoms;
R.sub.6 is Ile, Met, Nle or desR.sub.6 ;
R.sub.7 is Ser(Z.sub.1), Ala, Agl(Z.sub.2)(Z.sub.3), or MeAgl(Z.sub.2)(Z.sub.3);
Z.sub.1 is H or OCH.sub.3 ;
Z.sub.2 is H or lower alkyl;
Z.sub.3 is H or an acyl group having up to 7 carbon atoms;
R.sub.8 is Leu or Ile;
R.sub.10 is Leu or CML;
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, CML, Nle or Met;
R.sub.19 is CML, Leu or Ile;
R.sub.20 is Glu, D-Glu, Cys or His;
R.sub.21 is Nle, Leu, CML or Met;
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, CML 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 Aib or an L-isomer of a natural .alpha.-amino acid other than Cys;
R.sub.32 is His, D-His, Aib or an L- or D-isomer .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, CML or Leu;
R.sub.37 is CML, Leu, Nle or Tyr;
R.sub.38 is Nle, Met, CML or Leu;
R.sub.39 is Glu, Aib or Asp;
R.sub.40 is Ile, Aib, CML, Thr, Glu, Ala, Val, Leu, Nle, Phe, Nva, Gly or Gln;
and R.sub.41 is Ala, Aib, Ile, CML, Gly, Val, Leu, Nle, Phe, Nva or Gln;
wherein D-Phe may be substituted by Phe, Leu, Tyr, D-Leu, D-Tyr, D-Cpa, D-Trp, D-Nal, D-Pal or another D-isomer .alpha.-amino acid;
provided that a second cyclizing bond may exist between R.sub.20 and R.sub.23.

35 more elements

A CRF agonist peptide having the formula:

(cyclo 30-33)Y.sub.1 -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 -Nle-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 an acyl group having not more than 7 carbon atoms;
wherein R.sub.6 is Ile, Met, Nle or desR.sub.6 ;
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 or CML;
R.sub.18 is Val, CML, Nle or Met;
R.sub.19 is Leu or CML;
R.sub.20 is D-Glu or Glu;
R.sub.22 is Ala, D-Ala, Aib, Thr, Asp or Glu;
R.sub.23 is Arg or Lys;
R.sub.24 is Ala or Aib;
R.sub.25 is Asp or Glu;
R.sub.26 is Gln, Asn or Lys;
R.sub.27 is Leu or CML;
R.sub.28 is Ala 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, 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, CML or Leu;
R.sub.37 is CML, Leu or Tyr;
R.sub.39 is Glu, Aib or Asp;
R.sub.40 is Ile, CML, Aib, Thr, Glu, Ala, Val, Leu, Nle, Phe, Nva, Gly or Gln;
and R.sub.41 is Ala, Aib, Ile, CML, Gly, Val, Leu, Nle, Phe, Nva or Gln;
wherein D-Leu or Phe or Leu may be substituted for D-Phe.

27 more elements

Dependencies are inferred from the claim text.

Cyclic CRF agonists

Life of this patent

Cyclic CRF agonists

Life of this patent

Abstract

Claims25