TH2175B - GRF analogues - Google Patents

Abstract

The characteristics and synthesis of GRF from human pancreas (phGRF), rat hypothalamus (rGRF), and porcine hypothalamus (pGRF) have been studied for a long time. This invention represents a synthetic peptide that exhibits potent activity in stimulating pituitary GH secretion in animals, including humans, and is resistant to enzyme degradation in the body. The peptide sequence is as follows: R1-R2-R3-Ala-Ile-Phe-Thr-R8-Ser-R10-Arg-R12-R13-Leu-R15-Gln-R17-R18-Ala-Arg-Lys-Leu-R23-R24-R25Ile-R27-R28-Arg-Gln-Gly-Glu-R34-Asn-Gln-Glu-R38-R39-R40-Arg-R42-R43-R44 where R1 is Tyr-D-Tyr,Met,Phe,D-Phe,pCl-Phe-Leu,His or D-His, which has C (alpha) Me or N (alpha)Me substitution or no substitution; R2 is Ala or D-Ala; R3 is Asp or D-Asp; R8 is Ser, Asn, D-Ser or D-Asn R10 is Tyr or D-Tyr; R12 is Arg or Lys; R13 is Ile or Val; R15 is Gly or D-Ala; R17 is Leu or D-Leu; R18 is Tyr or Ser; R23 is Leu or D-Leu; R24 is His or Gln; R25 is Glu, Asp, D-Glu or D-Asp; R327 is Met, D-Met, Ala, Nle, Ile, Leu, Nva or Val; R28 is Ser; R34 is Arg or Ser; R38 is Gln or Arg; R39 is Arg or Gly; R40 is Ser or Ala; R42 is Phe, Ala or Aal; R43 is Asn or Arg; R44 is a naturally occurring amino acid. Peptides are also available with (a) R1 having a C (alpha) Me or N (alpha) Me substitution, or (b) R17 and/or R23 being D-Leu, or (c) R25, possibly D-Glu or D-Asp. Peptides with one or all radicals from R29 to R44 missing or with their non-toxic salts are also available. The carbon ends of these peptides are commonly given in amide form. These peptides and their non-toxic salts can be administered to animals, including humans and cold-blooded animals, to stimulate CH secretion or used in disease diagnosis.

Claims (12)

1. The process for producing peptides with this sequence. R1-R2-R3-Ala-Ile-Phe-Thr-R8-Ser-R10-Arg-R12-R13-Ler-R15-Gln-R17--R18-Ala-Arg-Lys-Leu-R23-R24-R25 Ile-R27-R28-Arg-Gln-Gly-Glu-R34 -Asn-Gln-Glu-R38-R39-R40-Arg-R42-R43-R44 where R1 is Tyr-D-Tyr,Met,Phe,D-Phe,pCl-Phe-Leu,His or D-His, with substitution of C (alpha) Me or N (alpha)Me or no substitution; R2 is Ala or D-Ala; R3 is Asp or D-Asp; R8 is Ser, Asn, D-Ser or D-Asn R10 is Tyr or D-Tyr; R12 is Arg or Lys; R13 is Ile or Val; R15 is Gly or D-Ala; R17 is Leu or D-Leu; R18 is Tyr or Ser; R23 is Leu or D-Leu; R24 is His or Gln; R25 is Glu, Asp, D-Glu or D-Asp; R327 is Met, D-Met, Ala, Nle, Ile, Leu, Nva or Val; R28 is Ser; R34 is Arg or Ser; R38 is Gln or Arg; R39 is Arg or Gly; R40 is Ser or Ala; R42 is Phe, Ala or Aal; R43 is Asn or Arg; R44 is a naturally occurring amino acid. and to form a peptide in which (a) R1 is replaced by C (alpha) Me or N (alpha) Me or (b) R17 and/or R23 is D-Leu or (c) R25, may be D-Glu or D-Asp and also to form a peptide in which one or all radicals between R29 and R44 are missing, which consists of (i) forming a compound that has at least one protective group and has the formula (II) X1-R1(X or X2)-R2-R3(X3)-Ala-Ile-Phe-Thr (X4)-R8 (X4 or X5)-Ser (X4)-R10(X2)-Arg(X6)-R12(X6 or X7)-R13-Leu-R15-(Gln(X5)-R17-R18(X2)-Ala-Arg(X6)-Lys(X7)-Leu-R23R24(X5 or X5)-R25(X3)-Ile-R27-R28(X4 or X5)-Arg(X6)-Gln(X5)-Gln(X5)-Gly-Glu(X3)-R34-(X4- or X6)-Asn(X5)-Gln(X5)-Glu(X3)-R38(X5 or X6)-R39(X6)-R40(X4)- where(X),(X1),(X2),(X3),(X4),(X5), (X6),(X7) and (X8) are all hydrogen or protective groups and where (X9) may be an amide or hydroxyl group or a protective group and (ii) isolate the protective group from the compound with formula II and give a peptide with formula (I) or its fragments that are biologically active and, if desired, (iii) convert the resulting peptide into its non-toxic salts. 2. The process described in claim 1, where R25 is D-Glu. 3. The procedure described in claim 1, where R25 is D-Asp. 4. The procedure described in claims 1, 2, or 3, where R23 is D-Leu. 5. Any of the procedures mentioned in claims 1-4, where R23 is D-Leu. 6. Any of the procedures mentioned in claims 1-5, where R27 is Nle. 7. Any of the procedures described in Claims 1-6, where R1 replaces C(alpha)Me or N(alpha)Me. 8. Any of the procedures outlined in Claims 1-7 shall be used, where X1 is hydrogen or protective (alpha)-amino group; X2 is hydrogen or protective phenolic hydroxyapatite of Tyr; X3 is hydrogen or protective carboxypatite of Asp or Glu; X4 is hydrogen or protective alcoholic hydroxyapatite of Thr; X5 is hydrogen or protective alcoholic hydroxyapatite of Ser; X6 is hydrogen or protective nidinopatite of Arg; R7 is hydrogen or protective amino side-chain of Lys; X8 is hydrogen or protective appropriate side-chain group; X is hydrogen or protective imidazole nitrogen group of His; and X9 is selected from groups consisting of OH, OCH3, ester, amide, hydrazide, -OCH2-supporting resin, and -NH-supporting resin. 9. The process described in claim 8, where X1 is BOC, X2 is 2,6-dichlorobenzyl, X3 is Bzl, X4 is Bzl, X5 is Bzl, X6 is Tos, X7 is 2-chlorobenzyl oxycarbonyl, and X9 is -O-CH2- supporting resin. 10. The process described in claim 8, where X1 is BOC, X2 is 2,6-dichlorobenzyl, X3 is Bzl, X4 is Bzl, X5 is Bzl, X6 is Tos, X7 is 2-chlorobenzyl oxycarbonyl and X9 is -NH- supporting resin. 11. An approach to improving agriculture by stimulating the release of GH in non-human warm-blooded animals to increase muscle mass and/or milk production using peptides or salts as defined in Representation 1 in effective doses. 12. An agricultural improvement method in which the release of GH in cold-blooded animals is stimulated to accelerate their growth by the use of peptides or salts as defined in Representation 1 in effective quantities.