Claims
- 1. A composition effective to cause the release and elevation of the level of growth hormone in the blood of an animal, the composition comprising an effective amount of polypeptides selected from at least two different groups of Group 1 polypeptides, Group 2 polypeptides or Group 3 polypeptides. In a ratio such that such composition is effective to cause the synergistic release and elevation of growth hormone in the blood of such animal;
- wherein Group 1 polypeptides are selected from any of the mutually occurring growth hormone releasing hormones and functional equivalents thereof, wherein said polypeptides act at the growth hormone releasing hormone receptor of mammals and other vertebrates, and crustaceans;
- Group 2 polypeptides are selected from any of the polypeptides having the structure:
- Ala-His-DTrp-Ala-Trp-DPhe-Lys-Gly-Tyr-NH.sub.2 ;
- Ala-His-(formyl)DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 (DTrp is formylated at the indole nitrogen);
- Ala-His-DTrp-Ser-Trp-DPhe-Lys-NH.sub.2 ;
- Cys-Ala-His-DTrp-Ala-Trp-DPhe-Lys-Cys-NH.sub.2 (cyclic disulfide);
- Cys-Ala-His-DTrp-Ala-Trp-DPhe-Lys-Cys-NH.sub.2 (free dithiol);
- DOPA-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- His-DTrp-Ala-Trp-DPhe-Lys-Ala-Tyr-NH.sub.2 ;
- Lys-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Lys-His-DTrp-Ala-Trp-DPhe-Asp-NH.sub.2 ;
- Phe-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Tyr-Ala-Gly-Thr-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Tyr-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Tyr-DAla-Gly-Phe-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Ala-His-XTrp*-Ala-Trp-DPhe-Lys-NH.sub.2 (*XTrp is selected from the group consisting of all N-monomethylated Trp isomers, i.e., (N.sup..alpha. Me)Trp, (N.sup..alpha. Me)DTrp, (indole NMe)Trp and (indole NMe)DTrp);
- Z-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- wherein Z is selected from the group consisting of all naturally occurring L-amino acids, Met(O), DOPA and Abu;
- and organic or inorganic addition salts of any of said polypeptides of Group 2; and
- Group 3 polypeptides are selected from any of the polypeptides having the structure:
- Tyr-DArg-Phe-NH.sub.2 ;
- Tyr-DAla-Phe-NH.sub.2 ;
- Tyr-DArg(NO.sub.2)-Phe-NH.sub.2 ;
- Tyr-DMet(O)-Phe-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-NH.sub.2 ;
- Tyr-DThr-Phe-Gly-NH.sub.2 ;
- Phe-DArg-Phe-Gly-NH.sub.2 ;
- Tyr-DArg-Phe-Sar;
- Tyr-DAla-Gly-Phe-NH.sub.2 ;
- Tyr-DArg-Gly-Trp-NH.sub.2 ;
- Tyr-DArg(NO.sub.2)-Phe-Gly-NH.sub.2 ;
- Tyr-DMet(O)-Phe-Gly-NH.sub.2 ;
- (NMe)Tyr-DArg-Phe-Sar-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-ol;
- Tyr-DArg-Gly-(NMe)Phe-NH.sub.2 ;
- Try-DArg-Phe-Sar-ol
- Tyr-DAla-Phe-Sar-ol
- Tyr-DAla-Phe-Gly-Tyr-NH.sub.2 ;
- Gly-Tyr-DArg-Phe-Gly-NH.sub.2 ;
- Tyr-DThr-Gly-Phe-Thz-NH.sub.2 ;
- Gly-Tyr-DAla-Phe-Gly-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-ol;
- Tyr-DAla-Gly-(NMe)Phe-Gly-ol;
- Tyr-DArg-Phe-Sar-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-NH.sub.2 ;
- Tyr-DAla-Phe-Sar;
- Tyr-DAla-Gly-(NMe)Phe-NH.sub.2 ;
- Sar-Tyr-DArg-Phe-Sar-NH.sub.2 ;
- Tyr-DCys-Phe-Gly-DCys-NH.sub.2 (cyclic disulfide);
- Tyr-DCys-Phe-Gly-DCys-NH.sub.2 (free dithiol);
- Tyr-DCys-Gly-Phe-DCys-NH.sub.2 (cyclic disulfide);
- Tyr-DCys-Gly-Phe-DCys-NH.sub.2 (free dithiol);
- Tyr-DAla-Phe-Gly-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Phe-Pro-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Phe-Hyp-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Sar-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Sar-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-Tyr-Pro-Ser-NH.sub.2 ; and organic or inorganic addition salts of any of said polypeptides of Group 3.
- 2. Composition of claim 1 wherein said Group 1 polypeptides are selected from any of the polypeptides:
- (a) having the following amino acid sequences in positions 1-44 (numbered from N terminus to C terminus):
- (#144) YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGESNQERGARARL-X,
- (#145) YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGERNQEQGARVRL-X,
- (#146) YADAIFTNSYRKVLGQLSARKLLQDIMNRQQGERNQEQGAKVRL-X,
- (#148) YADAIFTNSYRKILGQLSARKLLQDIMNRQQGERNQEQGAKVRL-X,
- (#149) HADAIFTSSYRRILGQLYARKLLHEIMNRQQGERNQEQRSRFN-X; and functional equivalents thereof;
- wherein the C-terminal amino acid has the following truncated general formula: ##STR3## wherein each R' independently represents the substituents of the particular amino acid residue, e.g.; hydrogen, alkyl, aryl, amino or acid substituents; X denotes the C terminal end group and is selected from --CONH.sub.2, --COOH, --COOR, --CONRR, --CH.sub.2 OH, and --CH.sub.2 OR, where R is an alkyl group having 1-6 carbon atoms or an aromatic ring having up to 12 carbon atoms; and wherein the amino acid residue abbreviations used are in accordance with the standard peptide nomenclature:
- G=Gly (Glycine),
- Y=Tyr (L-Tyrosine),
- I=Ile (L-Isoleucine),
- E=Glu (L-Glutamic Acid),
- T=Thr (L-Threonine),
- F=Phe (L-Phenylalanine),
- A=Ala (L-Alanine),
- K=Lys (L-Lysine),
- D=Asp (L-Aspartic Acid),
- C=Cys (L-Cysteine),
- R=Arg (L-Arginine),
- Q=Gln (L-Glutamine),
- P=Pro (L-Proline),
- L=Leu (L-Leucine),
- M=Met (L-Methionine),
- S=Ser (L-Serine),
- N=Asn (L-Asparagine),
- H=His (L-Histidine),
- W=Trp (L-Tryptophan), and
- V=Val (L-Valine);
- (b) any one of said (a) polypeptides having the following amino acid substitutions:
- position 1 of (#144-#148) is DTyr or His;
- position 1 of (#149) is Tyr or DHis;
- position 2 of (#144-#149) is (NMe)DAla or Aib or DAla;
- position 3 of (#144-#149) is DAsp;
- position 4 of (#144-#149) is DAla; and
- position 1+2 of (#144-#149) is;
- DTyr.sup.1 +DAla.sup.2, DTyr.sup.1 +(NMe)DAla.sup.2, or DTyr.sup.1 +Aib.sup.2 ;
- (c) any one of said (a) or (b) polypeptides having a substitution of Nle for Met at position 27;
- (d) any one of said (a), (b) or (c) polypeptides in which the N-terminus --NH.sub.2 is replaced by --NHCOR and wherein R is an alkyl group having 1 to 6 carbon atoms, or an aromatic ring having up to 12 carbon atoms;
- (e) fragments of any one of said (a), (b), (c) or (d) polypeptides which contain at least the amino acid residues of positions 1-29;
- (f) having the following specific amino acid sequences in positions 1-29 (numbered from N terminus to C terminus):
- YADAIFTNSYRKVLQQLAARKLLQDIMSR-X,
- YADAIFTNSYRKVLQQLLARKLLQDIMSR-X,
- YSDAIFSNAYRKILQQLLARKLLQDIMQR-X,
- YADAIFSNAYRKILQQLLARKLLQDIMQR-X,
- YADAIFSSAYRRLLAQLASRRLLQELLAR-X,
- YADAIFTNCYRKVLCQLSARKLLQDIMSR-X (linear dithiol), and
- YADAIFTNCYRKVLCQLSARKLLQDIMSR-X (cyclic disulfide);
- wherein the C-terminal amino acid and X are as defined above; and modification of any one of these group (f) compounds in accordance with the modifications set forth in (b), (c) and (d) above; and
- (g) organic or inorganic addition salts of any of said (a), (b), (c), (d), (e) or (f) polypeptides of Group 1.
- 3. A composition effective to cause the release and elevation of the level of growth hormone in the blood of an animal, the composition comprising an effective amount of polypeptides selected from at least two different groups of Group 1 polypeptides, Group 2 polypeptides or Group 3 polypeptides.
- wherein Group 1 polypeptides are selected from any polypeptides:
- (a) having the following amino acid sequences in positions 1-44 (numbered from N terminus to C terminus):
- (#144) YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGESNQERGARARL-CONH.sub.2 (hGHRH),
- (#145) YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGERNQEQGARVRL-CONH.sub.2 (pGHRH),
- (#146) YADAIFTNSYRKVLGQLSARKLLQDIMNRQQGERNQEQGAKVRL-CONH.sub.2 (bGHRH),
- (#148) YADAIFTNSYRKILGQLSARKLLQDIMNRQQGERNQEQGAKVRL-CONH.sub.2 (oGHRH), and
- (#149) HADAIFTSSYRRILGQLYARKLLHEIMNRQQGERNQEQRSRFN-COOH (rGHRH);
- wherein the C-terminal amino acid has the following truncated general formula: ##STR4## wherein each R' independently represents the substituents of the particular amino acid residue, e.g.; hydrogen, alkyl, aryl, amino or acid substituents; X denotes the C terminal end group and is selected from --CONH.sub.2, --COOH, --COOR, --CONRR, --CH.sub.2 OH, and --CH.sub.2 OR, where R is an alkyl group having 1-6 carbon atoms or an aromatic ring having up to 12 carbon atoms; and wherein the amino acid residue abbreviations used are in accordance with the standard peptide nomenclature:
- G=Gly (Glycine),
- Y=Tyr (Tyrosine),
- I=Ile (L-Isoleucine),
- E=Glu (L-Glutamic Acid),
- T=Thr (L-Threonine),
- F=Phe (L-Phenylalanine),
- A=Ala (L-Alanine),
- K=Lys (L-Lysine),
- D=Asp (L-Aspartic Acid),
- C=Cys (L-Cysteine),
- R=Arg (L-Arginine),
- Q=Gln (L-Glutamine),
- P=Pro (L-Proline),
- L=Leu (L-Leucine),
- M=Met (L-Methionine),
- S=Ser (L-Serine),
- N=Asn (L-Asparagine),
- H=His (L-Histidine),
- W=Trp (L-Tryptophan), and
- V=Val (L-Valine);
- (b) any one of said (a) polypeptides having the following amino acid substitutions:
- position 1 of (#144-#148) is DTyr or His;
- position 1 of (#149) is Tyr;
- position 2 of (#144-#149) is (NMe)DAla or Aib or DAla;
- position 3 of (#144-#149) is DAsp;
- position 4 of (#144-#149) is DAla;
- position 1+2 of (#144-#149) is DTyr.sup.1 +DAla.sup.2 ;
- (c) any one of said (a) or (b) polypeptides having a substitution of Nle for Met at position 27;
- (d) any one of said (a), (b), or (c) polypeptides in which the N-terminus --NH.sub.2 is replaced by --NHCOCH.sub.3 ;
- (e) fragments of any one of said (a), (b), (c) or (d) polypeptides which contain at least the amino acid residues of positions 1-29,
- (f) having the following amino acid sequences in positions 1-29 (numbered from N terminus to C terminus):
- YADAIFTNSYRKVLQQLAARKLLQDIMSR-X,
- YADAIFTNSYRKVLQQLLARKLLQDIMSR-X,
- YSDAIFSNAYRKILQQLLARKLLQDIMQR-X,
- YADAIFSNAYRKILQQLLARKLLQDIMQR-X,
- YADAIFSSAYRRLLAQLASRRLLQELLAR-X,
- YADAIFTNCYRKVLCQLSARKLLQDIMSR-X (linear dithiol), and
- YADAIFTNCYRKVLCQLSARKLLQDIMSR-X (cyclic disulfide);
- wherein the C-terminal amino acid and X are as defined above; and modification of any one of these group (f) compounds in accordance with the modifications set forth in (b), (c) and (d) above; and
- (g) organic or inorganic addition salts of any of said (a), (b), (c), (d), (e) or (f) polypeptides of Group 1;
- Group 2 polypeptides are selected from any of polypeptides having the structure:
- His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- DOPA-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Phe-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Ala-His-(formyl)DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 (DTrp is formylated at the indole nitrogen);
- Ala-His-DTrp-Ser-Trp-DPhe-Lys-NH.sub.2 ;
- His-DTrp-Ala-Trp-DPhe-Lys-Ala-Tyr-NH.sub.2 ;
- Lys-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Tyr-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Lys-His-DTrp-Ala-Trp-DPhe-Asp-NH.sub.2 ; and
- Z-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- wherein Z is selected from the group consisting of all naturally occurring L-amino acids, Met(O), DOPA and Abu; and
- organic or inorganic addition salts of any of said polypeptides of Group 2; and
- Group 3 polypeptides are selected from any of polypeptides having the structure:
- Tyr-DArg-Phe-NH.sub.2 ;
- Tyr-DArg(NO.sub.2)-Phe-NH.sub.2 ;
- Tyr-DMet(O)-Phe-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-NH.sub.2 ;
- Tyr-DArg-Phe-Sar;
- Tyr-DAla-Gly-Phe-NH.sub.2 ;
- Tyr-DAla-Gly-(NMe)Phe-Gly-ol;
- Tyr-DAla-Phe-Sar-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-NH.sub.2 ;
- Tyr-DArg(NO.sub.2)-Phe-Gly-NH.sub.2 ;
- Tyr-DMet(O)-Phe-Gly-NH.sub.2 ;
- (NMe)Tyr-DArg-Phe-Sar-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-ol;
- Tyr-DAla-Phe-Gly-Tyr-NH.sub.2 ;
- Gly-Tyr-DArg-Phe-Gly-NH.sub.2 ;
- Gly-Tyr-DAla-Phe-Gly-NH.sub.2 ;
- Sar-Tyr-DArg-Phe-Sar-NH.sub.2 ;
- Tyr-DCys-Phe-Gly-DCys-NH.sub.2 (cyclic disulfide);
- Tyr-DCys-Phe-Gly-DCys-NH.sub.2 (free dithiol)
- Tyr-DCys-Gly-Phe-DCys-NH.sub.2 (cyclic disulfide);
- Tyr-DCys-Gly-Phe-DCys-NH.sub.2 (free dithiol);
- Tyr-DAla-Phe-Gly-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Sar-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-Darg-Phe-Sar-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-Tyr-Pro-Ser-NH.sub.2 ;
- and organic or inorganic addition salts of any of said polypeptides of Group 3.
- 4. A composition effective to cause the release and elevation of the level of growth hormone in the blood of a mammal, the composition comprising an effective amount of polypeptides selected from at least two different groups of Group 1 polypeptides, Group 2 polypeptides or Group 3 polypeptides.
- wherein Group 1 polypeptides are selected from any of the polypeptides:
- (a) having the following amino acid sequences in positions 1-44 (numbered from N terminus to C terminus):
- (#144) YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGESNQERGARARL-CONH.sub.2 (hGHRH),
- (#145) YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGERNQEQGARVRL-CONH.sub.2 (pGHRH),
- (#146) YADAIFTNSYRKVLGQLSARKLLQDIMNRQQGERNQEQGAKVRL-CONH.sub.2 (bGHRH),
- (#148) YADAIFTNSYRKILGQLSARKLLQDIMNRQQGERNQEQGAKVRL-CONH.sub.2 (oGHRH), and
- (#149) HADAIFTSSYRRILGQLYARKLLHEIMNRQQGERNQEQRSRFN-COOH (rGHRH);
- wherein the C-terminal amino acid has the following truncated general formula: ##STR5## wherein each R' independently represents the substituents of the particular amino acid residue, e.g.; hydrogen, alkyl, aryl, amino or acid substituents; X denotes the C terminal end group and is selected from --CONH.sub.2, --COOH, --COOR, --CONRR, --CH.sub.2 OH, and --CH.sub.2 OR, where R is an alkyl group having 1-6 carbon atoms or an aromatic ring having up to 12 carbon atoms; and wherein the amino acid residue abbreviations used are in accordance with the standard peptide nomenclature:
- G=Gly (Glycine),
- Y=Tyr (Tyrosine),
- I=Ile (L-Isoleucine),
- E=Glu (L-Glutamic Acid),
- T=Thr (L-Threonine),
- F=Phe (L-Phenylalanine),
- A=Ala (L-Alanine),
- K=Lys (L-Lysine),
- D=Asp (L-Aspartic Acid),
- C=Cys (L-Cysteine),
- R=Arg (L-Arginine),
- Q=Gln (L-Glutamine),
- P=Pro (L-Proline),
- L=Leu (L-Leucine),
- M=Met (L-Methionine),
- S=Ser (L-Serine),
- N=Asn (L-Asparagine),
- H=His (L-Histidine),
- W=Trp (L-Tryptophan), and
- V=Val (L-Valine);
- (b) any one of said (a) polypeptides having a substitution of Nle for Met at position 27;
- (c) any one of said (a) or (b) polypeptides in which the N-terminus --NH.sub.2 is replaced by --NHCOCH.sub.3 ;
- (d) fragments of any one of said (a), (b) or (c) polypeptides which contain at least the amino acid residues of positions 1-29,
- (e) having the following amino acid sequences in positions 1-29 (numbered from N terminus to C terminus):
- YADAIFTNSYRKVLQQLAARKLLQDIMSR-X,
- YADAIFTNSYRKVLQQLLARKLLQDIMSR-X,
- YSDAIFSNAYRKILQQLLARKLLQDIMQR-X,
- YADAIFSNAYRKILQQLLARKLLQDIMQR-X,
- YADAIFSSAYRRLLAQLASRRLLQELLAR-X,
- YADAIFTNCYRKVLCQLSARKLLQDIMSR-X (linear dithiol), and
- YADAIFTNCYRKVLCQLSARKLLQDIMSR-X, (cyclic disulfide);
- wherein the C-terminal amino acid and X are as defined above; and modification of any one of these group (e) compounds in accordance with the modifications set forth in (b), (c) and (d) above;
- (f) organic or inorganic addition salts of any of said (a), (b), (c), (d) or (e) polypeptides of Group 1;
- Group 2 polypeptides are selected from any of the polypeptides having the structure:
- His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- DOPA-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- His-DTrp-Ala-Trp-DPhe-Lys-Ala-Tyr-NH.sub.2 ;
- Lys-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Tyr-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Phe-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Z-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- wherein Z is selected from the group consisting of Ala, Val, DOPA, Trp, Met, Lys, Asp, Met(O), Leu, Abu and Arg, and
- organic or inorganic addition salts of any of said polypeptides of Group 2; and
- Group 3 polypeptides are selected from any of the polypeptides having the structure:
- Tyr-DArg-Phe-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-NH.sub.2 ;
- Tyr-DArg-Phe-Sar;
- Tyr-DAla-Gly-Phe-NH.sub.2 ;
- Tyr-DAla-Gly-(NMe)Phe-Gly-ol;
- Tyr-DArg-Phe-Sar-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-ol;
- Tyr-DArg(NO.sub.2)-Phe-Gly-NH.sub.2 ;
- Tyr-DMet(O)-Phe-Gly-NH.sub.2 ;
- (NMe)Tyr-DArg-Phe-Sar-NH.sub.2 ;
- Gly-Tyr-DArg-Phe-Gly-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-Tyr-Hyp-Ser-NH.sub.2 ;
- and organic or inorganic addition salts of any of said polypeptides of Group 3.
- 5. Composition of claim 1 comprising a compound from each of Group 1 polypeptides and Group 2 polypeptides.
- 6. Composition of claim 1 comprising a compound from each of Group 1 polypeptides and Group 3 polypeptides.
- 7. Composition of claim 1 comprising a compound from each of Group 2 polypeptides and Group 3 polypeptides.
- 8. Composition of claim 1 comprising a compound from each of Group 1 polypeptides, Group 2 polypeptides and Group 3 polypeptides.
- 9. Composition of claim 3 comprising a compound from each of Group 1 polypeptides and Group 2 polypeptides.
- 10. Composition of claim 3 comprising a compound from each of Group 1 polypeptides and Group 3 polypeptides.
- 11. Composition of claim 3 comprising a compound from each of Group 2 polypeptides and Group 3 polypeptides.
- 12. Composition of claim 3 comprising a compound from each of Group 1 polypeptides, Group 2 polypeptides and Group 3 polypeptides.
- 13. Composition of claim 4 comprising a compound from each of Group 1 polypeptides and Group 2 polypeptides.
- 14. Composition of claim 4 comprising a compound from each of Group 1 polypeptides and Group 3 polypeptides.
- 15. Composition of claim 4 comprising a compound from each of Group 2 polypeptides and Group 3 polypeptides.
- 16. Composition of claim 4 comprising a compound from each of Group 1 polypeptides, Group 2 polypeptides and Group 3 polypeptides.
- 17. Method of causing release and elevation of the level of growth hormone in the blood of an animal, comprising administering an effective dose of a combination comprising polypeptides selected from at least two different groups of Group 1 polypeptides in a ratio such that such combination is effective to cause the synergistic release and elevation of growth hormone in the blood of such animal; Group 2 polypeptides or Group 3 polypeptides.
- wherein Group 1 polypeptides are selected from any of the naturally occurring growth hormone releasing hormones and functional equivalents thereof, wherein said polypeptides act at the growth hormone releasing hormone receptor of mammals and other vertebrates, and crustaceans;
- Group 2 polypeptides are selected from any of the polypeptides having the structure:
- Ala-His-DTrp-Ala-Trp-DPhe-Lys-Gly-Tyr-NH.sub.2 ;
- Ala-His-(formyl)DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 (DTrp is formylated at the indole nitrogen);
- Ala-His-DTrp-Ser-Trp-DPhe-Lys-NH.sub.2 ;
- Cys-Ala-His-DTrp-Ala-Trp-DPhe-Lys-Cys-NH.sub.2 (cyclic disulfide);
- Cys-Ala-His-DTrp-Ala-Trp-DPhe-Lys-Cys-NH.sub.2 (free dithiol);
- DOPA-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- His-DTrp-Ala-Trp-DPhe-Lys-Ala-Tyr-NH.sub.2 ;
- Lys-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Lys-His-DTrp-Ala-Trp-DPhe-Asp-NH.sub.2 ;
- Phe-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Tyr-Ala-Gly-Thr-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Tyr-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Tyr-DAla-Gly-Phe-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Ala-His-XTrp*-Ala-Trp-DPhe-Lys-NH.sub.2 (*XTrp is selected from the group consisting of all N-monomethylated Trp isomers, i.e., (N.sup..alpha. Me)Trp, (N.sup..alpha. Me)DTrp, (indole NMe)Trp and (indole NMe)DTrp);
- Z-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- wherein Z is selected from the group consisting of all naturally occurring L-amino acids, Met(O), DOPA and Abu;
- and organic or inorganic addition salts of any of said polypeptides of Group 2; and
- Group 3 polypeptides are selected from any of the polypeptides having the structure:
- Tyr-DArg-Phe-NH.sub.2 ;
- Tyr-DAla-Phe-NH.sub.2 ;
- Tyr-DArg(NO.sub.2)-Phe-NH.sub.2 ;
- Tyr-DMet(O)-Phe-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-NH.sub.2 ;
- Tyr-DThr-Phe-Gly-NH.sub.2 ;
- Phe-DArg-Phe-Gly-NH.sub.2 ;
- Tyr-DArg-Phe-Sar;
- Tyr-DAla-Gly-Phe-NH.sub.2 ;
- Tyr-DArg-Gly-Trp-NH.sub.2 ;
- Tyr-DArg(NO.sub.2)-Phe-Gly-NH.sub.2 ;
- Tyr-DMet(O)-Phe-Gly-NH.sub.2 ;
- (NMe)Tyr-DArg-Phe-Sar-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-ol;
- Tyr-DArg-Gly-(NMe)Phe-NH.sub.2 ;
- Try-DArg-Phe-Sar-ol
- Try-DAla-Phe-Sar-ol
- Tyr-DAla-Phe-Gly-Tyr-NH.sub.2 ;
- Gly-Tyr-DArg-Phe-Gly-NH.sub.2 ;
- Tyr-DThr-Gly-Phe-Thz-NH.sub.2 ;
- Gly-Tyr-DAla-Phe-Gly-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-ol;
- Tyr-DAla-Gly-(NMe)Phe-Gly-ol;
- Tyr-DArg-Phe-Sar-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-NH.sub.2 ;
- Tyr-DAla-Phe-Sar;
- Tyr-DAla-Gly-(NMe)Phe-NH.sub.2 ;
- Sar-Tyr-DArg-Phe-Sar-NH.sub.2 ;
- Tyr-DCys-Phe-Gly-DCys-NH.sub.2 (cyclic disulfide);
- Tyr-DCys-Phe-Gly-DCys-NH.sub.2 (free dithiol);
- Tyr-DCys-Gly-Phe-DCys-NH.sub.2 (cyclic disulfide);
- Tyr-DCys-Gly-Phe-DCys-NH.sub.2 (free dithiol);
- Tyr-DAla-Phe-Gly-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Phe-Pro-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Phe-Hyp-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Sar-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Sar-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-Tyr-Pro-Ser-NH.sub.2 ; and organic or inorganic addition salts of any of said polypeptides of Group 3.
- 18. Method of claim 17 wherein said Group 1 polypeptides are selected from any of the polypeptides:
- (a) having the following amino acid sequences in positions 1-44 (numbered from N terminus to C terminus):
- (#144) YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGESNQERGARARL-X,
- (#145) YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGERNQEQGARVRL-X,
- (#146) YADAIFTNSYRKVLGQLSARKLLQDIMNRQQGERNQEQGAKVRL-X,
- (#148) YADAIFTNSYRKILGQLSARKLLQDIMNRQQGERNQEQGAKVRL-X,
- (#149) HADAIFTSSYRRILGQLYARKLLHEIMNRQQGERNQEQRSRFN-X; and functional equivalents thereof;
- wherein the C-terminal amino acid has the following truncated general formula: ##STR6## wherein each R' independently represents the substituents of the particular amino acid residue, e.g.; hydrogen, alkyl, aryl, amino or acid substituents; X denotes the C terminal end group and is selected from --CONH.sub.2, --COOH, --COOR, --CONRR, --CH.sub.2 OH, and --CH.sub.2 OR, where R is an alkyl group having 1-6 carbon atoms or an aromatic ring having up to 12 carbon atoms; and wherein the amino acid residue abbreviations used are in accordance with the standard peptide nomenclature:
- G=Gly (Glycine),
- Y=Tyr (L-Tyrosine),
- I=Ile (L-Isoleucine),
- E=Glu (L-Glutamic Acid),
- T=Thr (L-Threonine),
- F=Phe (L-Phenylalanine),
- A=Ala (L-Alanine),
- K=Lys (L-Lysine),
- D=Asp (L-Aspartic Acid),
- C=Cys (L-Cysteine),
- R=Arg (L-Arginine),
- Q=Gln (L-Glutamine),
- P=Pro (L-Proline),
- L=Leu (L-Leucine),
- M=Met (L-Methionine),
- S=Ser (L-Serine),
- N=Asn (L-Asparagine),
- H=His (L-Histidine),
- W=Trp (L-Tryptophan), and
- V=Val (L-Valine);
- (b) any one of said (a) polypeptides having the following amino acid substitutions:
- position 1 of (#144-#148) is DTyr or His;
- position 1 of (#149) is Tyr or DHis;
- position 2 of (#144-#149) is (NMe)DAla or Aib or DAla;
- position 3 of (#144-#149) is DAsp;
- position 4 of (#144-#149) is DAla; and
- position 1+2 of (#144-#149) is;
- DTyr.sup.1 +DAla.sup.2, DTyr.sup.1 +(NMe)DAla.sup.2, or DTyr.sup.1 +Aib.sup.2 ;
- (c) any one of said (a) or (b) polypeptides having a substitution of Nle for Met at position 27;
- (d) any one of said (a), (b) or (c) polypeptides in which the N-terminus --NH.sub.2 is replaced by --NHCOR and wherein R is an alkyl group having 1 to 6 carbon atoms, or an aromatic ring having up to 12 carbon atoms;
- (e) fragments of any one of said (a), (b), (c) or (d) polypeptides which contain at least the amino acid residues of positions 1-29;
- (f) having the following specific amino acid sequences in positions 1-29 (numbered from N terminus to C terminus);
- YADAIFTNSYRKVLQQLAARKLLQDIMSR-X,
- YADAIFTNSYRKVLQQLLARKLLQDIMSR-X,
- YSDAIFSNAYRKILQQLLARKLLQDIMQR-X,
- YADAIFSNAYRKILQQLLARKLLQDIMQR-X,
- YADAIFSSAYRRLLAQLASRRLLQELLAR-X,
- YADAIFTNCYRKVLCQLSARKLLQDIMSR-X (linear dithiol), and
- YADAIFTNCYRKVLCQLSARKLLQDIMSR-X (cyclic disulfide);
- wherein the C-terminal amino acid and X are as defined above; and modification of any one of these group (f) compounds in accordance with the modifications set forth in (b), (c) and (d) above; and
- (g) organic or inorganic addition salts of any of said (a), (b), (c), (d), (e) or (f) polypeptides of Group 1.
- 19. Method of causing release and elevation of the level of growth hormone in the blood of an animal, comprising administering an effective dose of a combination comprising polypeptides selected from at least two different groups of Group 1 polypeptides, Group 2 polypeptides or Group 3 polypeptides, in a ratio such that such composition is effective to cause the synergistic release and elevation of growth hormone in the blood of such animal; wherein Group 1 polypeptides are selected from any polypeptides:
- (a) having the following amino acid sequences in positions 1-44 (numbered from N terminus to C terminus):
- (#144) YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGESNQERGARARL-CONH.sub.2 (hGHRH),
- (#145) YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGERNQEQGARVRL-CONH.sub.2 (pGHRH),
- (#146) YADAIFTNSYRKVLGQLSARKLLQDIMNRQQGERNQEQGAKVRL-CONH.sub.2 (bGHRH),
- (#148) YADAIFTNSYRKILGQLSARKLLQDIMNRQQGERNQEQGAKVRL-CONH.sub.2 (oGHRH), and
- (#149) HADAIFTSSYRRILGQLYARKLLHEIMNRQQGERNQEQRSRFN-COOH (rGHRH);
- wherein the C-terminal amino acid has the following truncated general formula: ##STR7## wherein each R' independently represents the substituents of the particular amino acid residue, e.g.; hydrogen, alkyl, aryl, amino or acid substituents; X denotes the C terminal end group and is selected from --CONH.sub.2, --COOH, --COOR, --CONRR, --CH.sub.2 OH, and --CH.sub.2 OR, where R is an alkyl group having 1-6 carbon atoms or an aromatic ring having up to 12 carbon atoms; and wherein the amino acid residue abbreviations used are in accordance with the standard peptide nomenclature:
- G=Gly (Glycine),
- Y=Tyr (Tyrosine),
- I=Ile (L-Isoleucine),
- E=Glu (L-Glutamic Acid),
- T=Thr (L-Threonine),
- F=Phe (L-Phenylalanine),
- A=Ala (L-Alanine),
- K=Lys (L-Lysine),
- D=Asp (L-Aspartic Acid),
- C=Cys (L-Cysteine),
- R=Arg (L-Arginine),
- Q=Gln (L-Glutamine),
- P=Pro (L-Proline),
- L=Leu (L-Leucine),
- M=Met (L-Methionine),
- S=Ser (L-Serine),
- N=Asn (L-Asparagine),
- H=His (L-Histidine),
- W=Trp (L-Tryptophan), and
- V=Val (L-Valine);
- (b) any one of said (a) polypeptides having the following amino acid substitutions:
- position 1 of (#144-#148) is DTyr or His;
- position 1 of (#149) is Tyr;
- position 2 of (#144-#149) is (NMe)DAla or Aib or DAla;
- position 3 of (#144-#149) is DAsp;
- position 4 of (#144-#149) is DAla;
- position 1+2 of (#144-#149) is DTyr.sup.1 +DAla.sup.2 ;
- (c) any one of said (a) or (b) polypeptides having a substitution of Nle for Met at position 27;
- (d) any one of said (a), (b), or (c) polypeptides in which the N-terminus --NH.sub.2 is replaced by --NHCOCH.sub.3 ;
- (e) fragments of any one of said (a), (b), (c) or (d) polypeptides which contain at least the amino acid residues of positions 1-29,
- (f) having the following amino acid sequences in positions 1-29 (numbered from N terminus to C terminus):
- YADAIFTNSYRKVLQQLAARKLLQDIMSR-X,
- YADAIFTNSYRKVLQQLLARKLLQDIMSR-X,
- YSDAIFSNAYRKILQQLLARKLLQDIMQR-X,
- YADAIFSNAYRKILQQLLARKLLQDIMQR-X,
- YADAIFSSAYRRLLAQLASRRLLQELLAR-X,
- YADAIFTNCYRKVLCQLSARKLLQDIMSR-X (linear dithiol), and
- YADAIFTNCYRKVLCQLSARKLLQDIMSR-X (cyclic disulfide);
- wherein the C-terminal amino acid and X are as defined above; and modification of any one of these group (f) compounds in accordance with the modifications set forth in (b), (c) and (d) above; and
- (g) organic or inorganic addition salts of any of said (a), (b), (c), (d), (e) or (f) polypeptides of Group 1;
- Group 2 polypeptides are selected from any of polypeptides having the structure:
- His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- DOPA-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Phe-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Ala-His-(formyl)DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 (DTrp is formylated at the indole nitrogen);
- Ala-His-DTrp-Ser-Trp-DPhe-Lys-NH.sub.2 ;
- His-DTrp-Ala-Trp-DPhe-Lys-Ala-Tyr-NH.sub.2 ;
- Lys-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Tyr-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Lys-His-DTrp-Ala-Trp-DPhe-Asp-NH.sub.2 ; and
- Z-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- wherein Z is selected from the group consisting of all naturally occurring L-amino acids, Met(O), DOPA and Abu; and
- organic or inorganic addition salts of any of said polypeptides of Group 2; and
- Group 3 polypeptides are selected from any of polypeptides having the structure:
- Tyr-DArg-Phe-NH.sub.2 ;
- Tyr-DArg(NO.sub.2)-Phe-NH.sub.2 ;
- Tyr-DMet(O)-Phe-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-NH.sub.2 ;
- Tyr-DArg-Phe-Sar;
- Tyr-DAla-Gly-Phe-NH.sub.2 ;
- Tyr-DAla-Gly-(NMe)Phe-Gly-ol;
- Tyr-DAla-Phe-Sar-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-NH.sub.2 ;
- Tyr-DArg(NO.sub.2)-Phe-Gly-NH.sub.2 ;
- Tyr-DMet(O)-Phe-Gly-NH.sub.2 ;
- (NMe)Tyr-DArg-Phe-Sar-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-ol;
- Tyr-DAla-Phe-Gly-Tyr-NH.sub.2 ;
- Gly-Tyr-DArg-Phe-Gly-NH.sub.2 ;
- Gly-Tyr-DAla-Phe-Gly-NH.sub.2 ;
- Sar-Tyr-DArg-Phe-Sar-NH.sub.2 ;
- Tyr-DCys-Phe-Gly-DCys-NH.sub.2 (cyclic disulfide);
- Tyr-DCys-Phe-Gly-DCys-NH.sub.2 (free dithiol)
- Tyr-DCys-Gly-Phe-DCys-NH.sub.2 (cyclic disulfide);
- Tyr-DCys-Gly-Phe-DCys-NH.sub.2 (free dithiol);
- Tyr-DAla-Phe-Gly-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Sar-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-Darg-Phe-Sar-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-Tyr-Pro-Ser-NH.sub.2 ;
- and organic or inorganic addition salts of any of said polypeptides of Group 3.
- 20. Method of causing release and elevation of the level of growth hormone in the blood of a mammal, comprising administering an effective dose of a combination comprising polypeptides selected from at least two different groups of Group 1 polypeptides, Group 2 polypeptides or Group 3 polypeptides in a ratio such that such combination is effective to cause the synergistic release and elevation of growth hormone in the blood of such mammal; wherein Group 1 polypeptides are selected from any of the polypeptides:
- (a) having the following amino acid sequences in positions 1-44 (numbered from N terminus to C terminus):
- (#144) YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGESNQERGARARL-CONH.sub.2 (hGHRH),
- (#145) YADAIFTNSYRKVLGQLSARKLLQDIMSRQQGERNQEQGARVRL-CONH.sub.2 (pGHRH),
- (#146) YADAIFTNSYRKVLGQLSARKLLQDIMNRQQGERNQEQGAKVRL-CONH.sub.2 (bGHRH),
- (#148) YADAIFTNSYRKILGQLSARKLLQDIMNRQQGERNQEQGAKVRL-CONH.sub.2 (oGHRH), and
- (#149) HADAIFTSSYRRILGQLYARKLLHEIMNRQQGERNQEQRSRFN-COOH (rGHRH);
- wherein the C-terminal amino acid has the following truncated general formula: ##STR8## wherein each R' independently represents the substituents of the particular amino acid residue, e.g.; hydrogen, alkyl, aryl, amino or acid substituents; X denotes the C terminal end group and is selected from --CONH.sub.2, --COOH, --COOR, --CONRR, --CH.sub.2 OH, and --CH.sub.2 OR, where R is an alkyl group having 1-6 carbon atoms or an aromatic ring having up to 12 carbon atoms; and wherein the amino acid residue abbreviations used are in accordance with the standard peptide nomenclature:
- G=Gly (Glycine),
- Y=Tyr (L-Tyrosine),
- I=Ile (L-Isoleucine),
- E=Glu (L-Glutamic Acid),
- T=Thr (L-Threonine),
- F=Phe (L-Phenylalanine),
- A=Ala (L-Alanine),
- K=Lys (L-Lysine),
- D=Asp (L-Aspartic Acid),
- C=Cys (L-Cysteine),
- R=Arg (L-Arginine),
- Q=Gln (L-Glutamine),
- P=Pro (L-Proline),
- L=Leu (L-Leucine),
- M=Met (L-Methionine),
- S=Ser (L-Serine),
- N=Asn (L-Asparagine),
- H=His (L-Histidine),
- W=Trp (L-Tryptophan), and
- V=Val (L-Valine);
- (b) any one of said (a) polypeptides having a substitution of Nle for Met at position 27;
- (c) any one of said (a) or (b) polypeptides in which the N-terminus --NH.sub.2 is replaced by --NHCOCH.sub.3 ;
- (d) fragments of any one of said (a), (b) or (c) polypeptides which contain at least the amino acid residues of positions 1-29,
- (e) having the following amino acid sequences in positions 1-29 (numbered from N terminus to C terminus):
- YADAIFTNSYRKVLQQLAARKLLQDIMSR-X,
- YADAIFTNSYRKVLQQLLARKLLQDIMSR-X,
- YSDAIFSNAYRKILQQLLARKLLQDIMQR-X,
- YADAIFSNAYRKILQQLLARKLLQDIMQR-X,
- YADAIFSSAYRRLLAQLASRRLLQELLAR-X,
- YADAIFTNCYRKVLCQLSARKLLQDIMSR-X (linear dithiol), and
- YADAIFTNCYRKVLCQLSARKLLQDIMSR-X, (cyclic disulfide);
- wherein the C-terminal amino acid and X are as defined above; and modification of any one of these group (e) compounds in accordance with the modifications set forth in (b), (c) and (d) above;
- (f) organic or inorganic addition salts of any of said (a), (b), (c), (d) or (e) polypeptides of Group 1;
- Group 2 polypeptides are selected from any of the polypeptides having the structure:
- His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- DOPA-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- His-DTrp-Ala-Trp-DPhe-Lys-Ala-Tyr-NH.sub.2 ;
- Lys-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Tyr-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- Phe-Ala-His-DTrp-Ala-TRp-DPhe-Lys-NH.sub.2 ;
- Z-His-DTrp-Ala-Trp-DPhe-Lys-NH.sub.2 ;
- wherein Z is selected from the group consisting of Ala, Val, DOPA, Trp, Met, Lys, Asp, Met(O), Leu, Abu and Arg, and
- organic or inorganic addition salts of any of said polypeptides of Group 2; and
- Group 3 polypeptides are selected from any of the polypeptides having the structure:
- Tyr-DArg-Phe-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-NH.sub.2 ;
- Tyr-DArg-Phe-Sar;
- Tyr-DAla-Gly-Phe-NH.sub.2 ;
- Tyr-DAla-Gly-(NMe)Phe-Gly-ol;
- Tyr-DArg-Phe-Sar-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-ol;
- Tyr-DArg(NO.sub.2)-Phe-Gly-NH.sub.2 ;
- Tyr-DMet(O)-Phe-Gly-NH.sub.2 ;
- (NMe)Tyr-DArg-Phe-Sar-NH.sub.2 ;
- Gly-Tyr-DArg-Phe-Gly-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Sar-Tyr-Hyp-Ser-NH.sub.2 ;
- Tyr-DAla-Phe-Gly-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-Tyr-Pro-Ser-NH.sub.2 ;
- Tyr-DArg-Phe-Gly-Tyr-Hyp-Ser-NH.sub.2 ;
- and organic or inorganic addition salts of any of said polypeptides of Group 3.
- 21. Method of claim 17 wherein said combination comprises a compound from each of Group 1 polypeptides and Group 2 polypeptides.
- 22. Method of claim 17 wherein said combination comprises a compound from each of Group 1 polypeptides and Group 3 polypeptides.
- 23. Method of claim 17 wherein said combination comprises a compound from each of Group 2 polypeptides and Group 3 polypeptides.
- 24. Method of claim 17 wherein said combination comprises a compound from each of Group 1 polypeptides, Group 2 polypeptides and Group 3 polypeptides.
- 25. Method of claim 19 wherein said combination comprises a compound from each of Group 1 polypeptides and Group 2 polypeptides.
- 26. Method of claim 19 wherein said combination comprises a compound from each of Group 1 polypeptides and Group 3 polypeptides.
- 27. Method of claim 19 wherein said combination comprises a compound from each of Group 2 polypeptides and Group 3 polypeptides.
- 28. Method of claim 19 wherein said combination comprises a compound from each of Group 1 polypeptides, Group 2 polypeptides and Group 3 polypeptides.
- 29. Method of claim 20 wherein said combination comprises a compound from each of Group 1 polypeptides and Group 2 polypeptides.
- 30. Method of claim 20 wherein said combination comprises a compound from each of Group 1 polypeptides and Group 3 polypeptides.
- 31. Method of claim 20 wherein said combination comprises a compound from each of Group 2 polypeptides and Group 3 polypeptides.
- 32. Method of claim 20 wherein said combination comprises a compound from each of Group 1 polypeptides, Group 2 polypeptides and Group 3 polypeptides.
SUMMARY
This application is a continuation-in-part of copending S.N. 861,968, filed May 12, 1986, now abandoned.
1. Field of Invention
The invention herein described relates to novel methods to promote release, and to produce elevated growth hormone (GH) levels in the blood of animals; and to combinations of polypeptide compounds acting in a synergistic manner to promote release and to produce elevated growth hormone (GH) levels in the blood of animals.
2. Description of the Art
It has been established in the scientific literature that the elevation of growth hormone levels in mammals upon administration of GH-releasing compounds can lead to enhanced body weight and to enhanced milk production if sufficiently elevated GH levels occur upon administration (c.f., P. K. Baker, et al., J. Animal Science 59 (supplement 1), 220 (1984); W. J. Croom et al., J. Dairy Sci. 67 (supplement 1), 109 (1984); S. N. McCutcheon et al., J. Dairy Sci. 67, 2881 (1984)). Further, it is known that the elevation of growth hormone levels in mammals can be accomplished by application of known growth hormone releasing agents, such as endogenous growth hormone releasing hormones (GHRH's, #144-#149) included herein as members of the group designated as "Group 1 compounds" (c.f., P. Brazeau et al., Proc. Natl. Acad. Sci. 79, 7909 (1982), and M. O. Thorner et al., Lancet 1,24 (1983)). The elevation of growth hormone levels in mammals can also be accomplished by application of growth hormone releasing peptides (GRP's), some of which have been previously described and are included herein as members of the group designated as "Group 2 compounds" (c.f. C. Y. Bowers et al., Endocrinology 114, 1537 (1984), F. A. Momany et al., Endocrinology 114, 1531 (1984) and C. Y. Bowers, 7th International Congress of Endocrinology Abstracts, 464 (1984)). Antibodies to the endogenous growth hormone release inhibitor, somatostatin (SRIF) are also used to elevate GH levels. In the last case, growth hormone levels are elevated by removing the endogenous GH-release inhibitor (SRIF) before it reaches the pituitary, where it inhibits the release of GH (c.f. W. B. Wehrenberg et al., Endocrinology 115, 1218 (1984)). Finally, it has been shown that some compounds such as morphine (c.f. C. Rivier et al., Endocrinology 100, 238 (1977)) and other alkaloids (c.f. C. Y. Bowers, Endocrinology 117, 1441 (1985)) and DAla.sup.2, DLeu.sup.5 -enkephalinamide (c.f. E. L. Lien et al., FEBS Letters 88, 208 (1978)) also release growth hormone by acting on the hypothalamus. The Group 3 compounds described herein may also act at the hypothalamus.
It is an object of this invention to provide novel combinations of growth hormone releasing compounds which directly or indirectly produce elevated levels of growth hormone in the blood of animals. Such combinations of compounds have enhanced stabilities, solubilities, physiochemical properties and biological activities relative to growth hormone releasing compounds of the prior art.
It is another object of this invention to provide, by administration of a combination of active ingredients described in greater detail below, a method for elevating GH levels in animals for use in the pharmaceutical and animal products industries.
Yet another object of this invention is to provide a variety of each of three differently acting growth hormone releasing compounds, each of which can be combined in different ways and as part of different compositions, to allow optimization in the design of formulas for different utilities, such as in the enhancement of growth; and the improvement in milk, fur or wool production.
Each of the groups of agents referred to above and their analogs act to promote the release of growth hormone in animals through different mechanisms, thus, the synergistic effect of the combination of compounds described herein in elevating the GH levels in vivo is unusual and unexpected for the combination of compounds between groups, as described herein. Further, the magnitude of the synergistic GH response to an effective dose of the combination of compounds between groups is not found in any single compound within a group, at similar doses.
How the compounds described in this invention act to achieve their remarkable synergistic effect is not fully understood at the molecular level. Clearly one might expect to see additive effects from application of combinations of these compounds. However, the very important synergistic results and their utility will become apparent in light of the accompanying disclosure. Of novelty to this preparation is the discovery by the authors that the Group 1 compounds described herein act synergistically with Group 2 compounds, to promote the release of much more GH than equivalent amounts of the individual compounds within groups, when given alone. In addition, the authors have discovered that the Group 1 compounds described herein act synergistically with Group 3 compounds, to promote the release of much more GH than equivalent amounts of the individual compounds within groups, when given alone. The authors have also discovered that the Group 2 compounds described herein act synergistically with Group 3 compounds, to promote the release of much more GH than equivalent amounts of the individual components within groups, when given alone. Further, the authors have recently shown that specific Group 3 compounds, such as the naturally occurring dermorphins, hereinafter referred to as compounds #8801 and #8802, also act synergistically with both Group 1 and 2 compounds, such that a combination mixture of all three groups of compounds is a potent GH-releasing mixture.
The present invention provides combinations having synergistic GH-releasing effects. The combinations of this invention may be used to enhance blood GH levels in animals; enhance milk production in cows; enhance body growth in animals such as mammals (e.g., humans, sheep, bovines, and swine), as well as fish, fowl and crustaceans; and increase wool and/or fur production in mammals. The amount of body growth is dependent upon the sex and age of the animal species, quantity and identity of the growth hormone releasing compound being administered, and route of administration.
This invention is concerned with novel combinations of polypeptides acting in a synergistic manner to promote, the release of and thereby produce elevated growth hormone levels in the blood of animals. The invention is also concerned with methods for use of the novel composition.
In its broadest scope, the present invention provides a combination effective to cause the release and elevation of the level of growth hormone in the blood of an animal, the combination comprising an effective amount of polypeptides selected from at least two different groups of the Group 1 polypeptides, Group 2 polypeptides or Group 3 polypeptides hereinafter described in greater detail.
In accordance with this broadest scope of the present invention, the term "Group 1 polypeptides" is intended to include naturally occurring growth hormone releasing hormones, e.g., human, porcine, bovine, ovine and rat growth hormone releasing hormones, such as compounds #144-149 below, and functional analogs thereof. Such peptides act at the growth hormone releasing hormone receptor of mammals and other vertebrates, crustaceans and the like, to cause the release of growth hormone. Representative peptides included within this definition are selected from any of the polypeptides:
(a) having the following amino acid sequences in positions 1-44 (numbered from N terminus to C terminus):
While essentially stereochemically pure D or L amino acids are referred to throughout this specification, it is to be understood that mixtures of the D/L stereoisomers of the amino acid residues are also operable, while sometimes having a reduced level of biological activity as a function of the relative amount of the unspecified configuration which is present. Additional amino acid and peptide abbreviations which appear throughout the specification include:
(b) any one of said (a) polypeptides having the following amino acid substitutions:
(c) any one of said (a) or (b) polypeptides having a substitution of Nle for Met at position 27;
(d) any one of said (a), (b) or (c) polypeptides in which the N-terminus --NH.sub.2 is replaced by --NHCOR and wherein R is an alkyl group having 1 to 6 carbon atoms, or an aromatic ring having up to 12 carbon atoms;
(e) fragments of any one of said (a), (b), (c) or (d) polypeptides which contain at least the amino acid residues of positions 1-29;
(f) having the following specific amino acid sequences in positions 1-29 (numbered from N terminus to C terminus):
(g) organic or inorganic addition salts of any of said (a), (b), (c), (d), (e) or (f) polypeptides of Group 1.
Group 2 polypeptides contemplated within the broad scope of the present invention are selected from any of the polypeptides having the structure:
Group 3 polypeptides contemplated within the broad scope of the present invention are selected from any of the polypeptides having the structure:
In a preferred embodiment, the present invention provides a combination effective to cause the release and elevation of growth hormone in the blood of an animal, the combination comprising an effective amount of polypeptides selected from at least two different groups of the Group 1 polypeptides, Group 2 polypeptides or Group 3 polypeptides set forth below.
In accordance with this preferred embodiment of the present invention, the term "Group 1 polypeptides" is intended to include naturally occurring growth hormones, e.g., human, porcine, bovine, ovine and rat growth hormone releasing hormones, such as compounds #144-149 below, and functional analogs thereof, which polypeptides act at the growth hormone releasing hormone receptor of mammals and other vertebrates. Compounds within this definition are selected from any of the polypeptides:
(a) having the following amino acid sequences in positions 1-44 (numbered from N terminus to C terminus):
(b) any one of said (a) polypeptides having the following amino acid substitutions:
(c) any one of said (a) or (b) polypeptides having a substitution of Nle for Met at position 27;
(d) any one of said (a), (b), or (c) polypeptides in which the N-terminus --NH.sub.2 is replaced by --NHCOCH.sub.3 ;
(e) fragments of any one of said (a), (b), (c) or (d) polypeptides which contain at least the amino acid residues of positions 1-29,
(f) having the following amino acid sequences in positions 1-29 (numbered from N terminus to C terminus):
(g) organic or inorganic addition salts of any of said (a), (b), (c), (d), (e) or (f) polypeptides of Group 1.
Group 2 polypeptides contemplated within this preferred scope of the invention are selected from any of the polypeptides having the structure:
Group 3 polypeptides contemplated within this preferred scope of the invention are selected from any of the polypeptides having the structure:
In a most preferred embodiment, the present invention provides a combination effective to cause the release and elevation of growth hormone in the blood of a mammal, the combination comprising an effective amount of polypeptides selected from at least two different groups of the Group 1 polypeptides, Group 2 polypeptides or Group 3 polypeptides set forth below.
In accordance with this most preferred embodiment of the present invention, the term "Group 1 polypeptides" is intended to include naturally occurring growth hormones, e.g., human, porcine, bovine, ovine and rat growth hormone releasing hormones, such as compounds #144-149 below, and functional analogs thereof, which polypeptides act at the growth hormone releasing hormone receptor of mammals. Compounds within this definition are selected from any of the polypeptides:
(a) having the following amino acid sequences in positions 1-44 (numbered from N terminus to C terminus):
(b) any one of said (a) polypeptides having a substitution of Nle for Met at position 27;
(c) any one of said (a) or (b) polypeptides in which the N-terminus --NH.sub.2 is replaced by --NHCOCH.sub.3 ;
(d) fragments of any one of said (a), (b) or (c) polypeptides which contain at least the amino acid residues of positions 1-29, and
(e) having the following amino acid sequences in positions 1-29 (numbered from N terminus to C terminus):
(f) organic or inorganic addition salts of any of said (a), (b), (c), (d) or (e) polypeptides of Group 1.
Group 2 polypeptides contemplated within this most preferred embodiment of the present invention, are selected from any of the polypeptides having the structure:
Group 3 polypeptides contemplated within this most preferred embodiment of the present invention are selected from any of the polypeptides having the structure:
The invention is also concerned with a method of causing release and elevation of growth hormone in the blood of an animal, comprising administering an effective dose of a combination comprising polypeptides selected from at least two different groups of the Group 1 polypeptides, Group 2 polypeptides or Group 3 polypeptides hereinafter described in greater detail.
In accordance with the broad scope of this embodiment of the present invention, the term "Group 1 polypeptides" is intended to include naturally occurring growth hormone releasing hormones, e.g., human, porcine, ovine, and rat growth hormone releasing hormones, such as compounds #144-149 below, and functional analogs thereof. Such peptides act at the growth hormone releasing hormone receptor of mammals and other vertebrates. crustaceans and the like, to cause the release of growth hormone. Representative peptides included within this definition are selected from any of the polypeptides:
(a) having the following amino acid sequences in positions 1-44 (numbered from N terminus to C terminus):
(b) any one of said (a) polypeptides having the following amino acid substitutions;
(c) any one of said (a) or (b) polypeptides having a substitution of Nle for Met at position 27;
(d) any one of said (a), (b) or (c) polypeptides in which the N-terminus --NH.sub.2 is replaced by --NHCOR and wherein R is an alkyl group having 1 to 6 carbon atoms or an aromatic ring having up to 12 carbon atoms;
(e) fragments of any one of said (a), (b), (c) or (d) polypeptides which contain at least the amino acid residues of positions 1-29;
(f) having the following amino acid sequences in positions 1-29 (numbered from N terminus to C terminus):
(g) organic or inorganic addition salts of any of said (a), (b), (c), (d), (e) or (f) polypeptides of Group 1.
Group 2 polypeptides contemplated within the broad scope of this embodiment of the invention are selected from any of the polypeptides having the structure:
Group 3 polypeptides contemplated within the broad scope of this embodiment of the invention are selected from any of the polypeptides having the structure:
In a preferred embodiment the invention provides a method of causing release and elevation of growth hormone in the blood of an animal, comprising administering an effective dose of a combination comprising polypeptides selected from at least two different groups of the Group 1 polypeptides, Group 2 polypeptides or Group 3 polypeptides set forth below.
In accordance with this preferred embodiment of the invention method of causing release and elevation of growth hormone in the blood of a mammal or a vertebrate, the term "Group 1 polypeptides" is intended to include naturally occurring growth hormones, e.g., human, porcine, bovine, ovine and rat growth hormone releasing hormones, such as compounds #144-149 below, and functional analogs thereof, which polypeptides act at the growth hormone releasing hormone receptor of mammals and other vertebrates. Compounds within this definition are selected from any of the polypeptides:
(a) having the following amino acid sequences in positions 1-44 (numbered from N terminus to C terminus):
(b) any one of said (a) polypeptides having the following amino acid substitutions:
(c) any one of said (a) or (b) polypeptides having a substitution of Nle for Met at position 27;
(d) any one of said (a), (b), or (c) polypeptides in which the N-terminus --NH.sub.2 is replaced by --NHCOCH.sub.3 ; and
(e) fragments of any one of said (a), (b), (c) or (d) polypeptides which contain at least the amino acid residues of positions 1-29,
(f) having the following amino acid sequences in positions 1-29 (numbered from N terminus to C terminus):
(g) organic or inorganic addition salts of any of said (a), (b), (c), (d), (e) or (f) polypeptides of Group 1.
Group 2 polypeptides contemplated within the preferred embodiment of the invention method are selected from any of the polypeptides having the structure:
Group 3 polypeptides contemplated within this preferred embodiment of the invention method are selected from any of the polypeptides having the structure:
In a most preferred embodiment the invention provides a method of causing release and elevation of growth hormone in the blood of a mammal, comprising administering an effective dose of a combination comprising polypeptides selected from at least two different groups of the Group 1 polypeptides, Group 2 polypeptides or Group 3 polypeptides set forth below.
In accordance with this most preferred embodiment of the invention method, the term "Group 1 polypeptides" is intended to include naturally occurring growth hormones, e.g., human, porcine, bovine, ovine and rat growth hormone releasing hormones, such as compounds #144-149 below, and functional analogs thereof, which polypeptides act at the growth hormone releasing hormone receptor of mammals. Compounds within this definition are selected from any of the polypeptides:
(a) having the following amino acid sequences in positions 1-44 (numbered from N terminus to C terminus):
(b) any one of said (a) polypeptides having a substitution of Nle for Met at position 27;
(c) any one of said (a) or (b) polypeptides in which the N-terminus --NH.sub.2 is replaced by --NHCOCH.sub.3 ;
(d) fragments of any one of said (a), (b) or (c) polypeptides which contain at least the amino acid residues of positions 1-29;
(e) having the following amino acid sequences in position 1-29 (numbered from N terminus to C terminus):
(f) organic or inorganic addition salts of any of said (a), (b), (c), (d) or (e) polypeptides of Group 1.
Group 2 polypeptides contemplated within this most preferred embodiment of the invention method are selected from any of the polypeptides having the structure:
Group 3 polypeptides contemplated within this most preferred embodiment of the invention method are selected from any of the polypeptides having the structure:
The invention further provides specific combinations of polypeptides and methods of using such combinations.
The substances described in this invention can be synthesized according to the usual methods of solution and solid phase peptide chemistry, or by classical methods known in the art. The solid-phase synthesis is commenced from the C-terminal end of the peptide. A suitable starting material can be prepared, for instance, by attaching the required protected alpha-amino acid to a chloromethylated resin, a hydroxymethyl resin, a benzhydrylamine (BHA) resin, or a para-methyl-benzylhydrylamine (p-Me-BHA) resin. One such chloromethyl resin is sold under the tradename BIOBEADS SX-1 by Bio Rad Laboratories, Richmond, Calif. The preparation of the hydroxymethyl resin is described by Bodansky et al., Chem. Ind. (London) 38, 1597 (1966). The BHA resin has been described by Pietta and Marshall, Chem. Commn., 650 (1970) and is commercially available from Penninsula Laboratories, Inc., Belmont, Calif., or Beckman Instruments, Inc., Palo Alto, Calif., in the hydrochloric form thereof (BHA.HCl).
After the initial attachment, the alpha-amino protecting group can be removed by a choice of acidic reagents, including trifluoroacetic acid (TFA) or hydrochloric acid (HCl) solutions in organic solvents at room temperature. After removal of the alpha-amino protecting group, the remaining protected amino acids can be coupled stepwise in the desired order. Each protected amino acid can be generally reacted in about a 3-fold excess using an appropriate carboxyl group activator such as dicyclohexylcarbodiimide (DCC) in solution, for example, in methylene chloride (CH.sub.2 Cl.sub.2) or dimethylformamide (DMF) and mixtures thereof.
After the desired amino acid sequence has been completed, the desired peptide can be cleaved from the resin support by treatment with a reagent such as hydrogen fluoride (HF) which not only cleaves the peptide from the resin, but also cleaves most commonly used side-chain protecting groups. When a chloromethyl resin or hydroxymethyl resin is used, HF treatment results in the formation of the free peptide acid. When the BHA or p-Me-BHA resin is used, HF treatment results directly in free peptide amides.
The solid-phase procedure discussed above is well known in the art and has been described by Stewart and Young, Solid Phase Peptide Synthesis: (Freeman and Co., San Francisco, 1969).
Some of the well known solution methods which can be employed to synthesize the peptide moieties of the instant invention are set forth in Bodansky et al., Peptide Synthesis, 2nd Edition, John Wiley & Sons, New York, N.Y. 1976.
The present invention includes within its scope compositions comprising, as an active ingredient, at least two of the compounds, or analogs thereof, described herein, or addition salts thereof, or their pharmaceutically acceptable salts in association with a carrier, diluent, slow release matrix, or coating.
The organic or inorganic addition salts of the growth hormone releasing compounds and combinations thereof contemplated to be within the scope of the present invention include salts of such organic moieties as acetate, trifluoroacetate, oxalate, valerate, oleate, laurate, benzoate, lactate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthalate, and the like; and such inorganic moieties as Group I (i.e., alkali metal salts), Group II (i.e., alkaline earth metal salts) ammonium and protamine salts, zinc, iron, and the like with counterions such as the chloride, bromide, sulfate, phosphate and the like, as well as the organic moieties referred to above.
Pharmaceutically acceptable salts are preferred when administration to human subjects is contemplated. Such salts include the non-toxic alkali metal, alkaline earth metal and ammonium salts commonly used in the pharmaceutical industry including the sodium, potassium, lithium, calcium, magnesium, barium, ammonium and protamine salts which are prepared by methods well known in the art. The term also includes non-toxic acid addition salts which are generally prepared by reacting the compounds of this invention with a suitable organic or inorganic acid. Representative salts include the hydrochloride, hydrobromide, sulfate, bisulfate, acetate, oxalate, valerate, oleate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napsylate, and the like.
The invention will now be described in greater detail by reference to the following non-limiting examples.
US Referenced Citations (66)
Non-Patent Literature Citations (12)
| Entry |
| P. K. Baker et al., J. Animal Science 59 (Supplement 1), 220 (1984). |
| W. J. Croom et al., J. Dairy Sci. 67 (Supplement 1), 109 (1984). |
| S. N. McCutcheon et al., J. Dairy Sci. 67, 2881 (1984). |
| P. Brazeau et al., Proc. Natl. Acad. Sci. 79, 7909 (1982). |
| M. O. Thorner et al., Lancet 1,24 (1983). |
| C. Y. Bowers et al., Endocrinology 114, 1537 (1984). |
| F. A. Momany et al., Endocrinology 114, 1531 (1984). |
| C. Y. Bowers, 7th International Congress of Endocrinology Ab., 464 (1984). |
| W. B. Wehrenberg et al., Endocrinology 115, 1218 (1984). |
| C. Rivier et al., Endocrinology 100, 238 (1977). |
| C. Y. Bowers, Endocrinology 117, 1441 (1985). |
| E. L. Lien et al., FEBS Letters 88, 208 (1978). |
Continuation in Parts (1)
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Number |
Date |
Country |
| Parent |
861968 |
May 1986 |
|
Patent Grant
4880778
