EXENDIN-4 DERIVATIVES AS DUAL GLP1/GLUCAGON AGONISTS
20180185450 ยท 2018-07-05
Inventors
- Torsten Haack (Frankfurt am Main, DE)
- Michael Wagner (Frankfurt am Main, DE)
- Bernd Henkel (Frankfurt am Main, DE)
- Siegfried Stengelin (Frankfurt am Main, DE)
- Andreas Evers (Frankfurt am Main, DE)
- Martin Bossart (Frankfurt am Main, DE)
Cpc classification
A61P1/04
HUMAN NECESSITIES
A61K45/06
HUMAN NECESSITIES
Y02A50/30
GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
A61P9/10
HUMAN NECESSITIES
A61K2300/00
HUMAN NECESSITIES
A61K2300/00
HUMAN NECESSITIES
A61P1/16
HUMAN NECESSITIES
International classification
A61K45/06
HUMAN NECESSITIES
Abstract
The present invention relates to exendin-4 derivatives and their medical use, for example in the treatment of disorders of the metabolic syndrome, including diabetes and obesity, as well as reduction of excess food intake.
Claims
1-31. (canceled)
32. A method for treating type 1 diabetes in a patient in need thereof, the method comprising administering to said patient a therapeutically effective amount of the peptidic compound of formula (I):
R.sup.1ZR.sup.2(I) or a salt, or solvate thereof, or a pharmaceutical composition comprising a compound of formula (I) or a salt or solvate thereof as an active agent together with at least one pharmaceutically acceptable carrier, wherein Z is a peptide moiety having formula (II): TABLE-US-00020 (II) His-X2-X3-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln- X14-X15-X16-X17-X18-Ala-X20-X21-Phe-Ile-Glu-Trp- Leu-Lys-X28-X29-Gly-Pro-Ser-Ser-Gly-X35-Pro-Pro- Pro-X39-X40 wherein: X2 is an amino acid residue selected from Ser, D-Ser, and Aib, X3 is an amino acid residue selected from Gln, His, and -amino-functionalized Gln, wherein Gln is optionally functionalized in that an H of the -NH.sub.2 group is substituted by (C.sub.1-C.sub.4)-alkyl, X14 is an amino acid residue having a side chain with a functionalized NH.sub.2 group, wherein the functionalized NH.sub.2 side chain group is functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms independently selected from halogen, N, O, S, P, and combinations thereof, X15 is an amino acid residue selected from Glu and Asp, X16 is an amino acid residue selected from Ser, Glu, and Lys, X17 is an amino acid residue selected from Arg, Glu, Gln, Leu, Aib, and Lys, X18 is an amino acid residue selected from Arg, Ala, and Lys, X20 is an amino acid residue selected from Gln, Arg, Lys, His, Glu, and Aib, X21 is an amino acid residue selected from Asp, Leu, and Glu, X28 is an amino acid residue selected from Asn, Arg, Lys, Aib, Ser, Glu, Ala, and Asp, X29 is an amino acid residue selected from Gly, Ala, D-Ala, and Thr, X35 is an amino acid residue selected from Ala, Glu, Arg, and Lys, X39 is Ser or is absent, and X40 is absent or is an amino acid residue having a side chain with an NH.sub.2 group, wherein the NH.sub.2 side chain group is optionally functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms selected from halogen, N, O, S, P, and combinations thereof, R.sup.1 is the N-terminal group of the peptidic compound and is selected from NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.5)alkyl], N[(C.sub.1-C.sub.5)alkyl].sub.2, NH[(C.sub.0-C.sub.4)alkylene-(C.sub.3-C.sub.8)cycloalkyl], NHC(O)H, NHC(O)(C.sub.1-C.sub.5)-alkyl, and NHC(O)(C.sub.0-C.sub.3)alkylene-(C.sub.3-C.sub.8)cycloalkyl, in which alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or halogen selected from F, Cl, Br, and I, R.sup.2 is the C-terminal group of the peptidic compound and is selected from (i) OH and functionalized OH, wherein the functionalized OH is selected from O(C.sub.1-C.sub.20)alkyl and O(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl, and (ii) NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.30)alkyl], N[(C.sub.1-C.sub.30)alkyl].sub.2, NH[(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl], N[C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl].sub.2, NH[(CH.sub.2CH.sub.2O).sub.1-40(C.sub.1-C.sub.4)alkyl], NH(C.sub.3-C.sub.8)heterocyclyl or NH(C.sub.0-C.sub.8)alkylene-aryl, wherein aryl is selected from phenyl or naphthyl, the (C.sub.3-C.sub.8)-heterocyclyl contains one N-atom and optionally two additional heteroatoms selected from O, N and S, and alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or a halogen selected from F, Cl, Br, and I.
33. The method according to claim 32, wherein said peptidic compound of formula (I) is any one of SEQ ID NOs: 4-181, 196-223, 226-229, or a salt or solvate thereof.
34. A method for treating type 2 diabetes in a patient in need thereof, the method comprising administering to said patient a therapeutically effective amount of the peptidic compound of formula (I):
R.sup.1ZR.sup.2(I) or a salt, or solvate thereof, or a pharmaceutical composition comprising a compound of formula (I) or a salt or solvate thereof as an active agent together with at least one pharmaceutically acceptable carrier, wherein Z is a peptide moiety having formula (II): TABLE-US-00021 (II) His-X2-X3-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln- X14-X15-X16-X17-X18-Ala-X20-X21-Phe-Ile-Glu-Trp- Leu-Lys-X28-X29-Gly-Pro-Ser-Ser-Gly-X35-Pro-Pro- Pro-X39-X40 wherein: X2 is an amino acid residue selected from Ser, D-Ser, and Aib, X3 is an amino acid residue selected from Gln, His, and -amino-functionalized Gln, wherein Gln is optionally functionalized in that an H of the -NH.sub.2 group is substituted by (C.sub.1-C.sub.4)-alkyl, X14 is an amino acid residue having a side chain with a functionalized NH.sub.2 group, wherein the functionalized NH.sub.2 side chain group is functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms independently selected from halogen, N, O, S, P, and combinations thereof, X15 is an amino acid residue selected from Glu and Asp, X16 is an amino acid residue selected from Ser, Glu, and Lys, X17 is an amino acid residue selected from Arg, Glu, Gln, Leu, Aib, and Lys, X18 is an amino acid residue selected from Arg, Ala, and Lys, X20 is an amino acid residue selected from Gln, Arg, Lys, His, Glu, and Aib, X21 is an amino acid residue selected from Asp, Leu, and Glu, X28 is an amino acid residue selected from Asn, Arg, Lys, Aib, Ser, Glu, Ala, and Asp, X29 is an amino acid residue selected from Gly, Ala, D-Ala, and Thr, X35 is an amino acid residue selected from Ala, Glu, Arg, and Lys, X39 is Ser or is absent, and X40 is absent or is an amino acid residue having a side chain with an NH.sub.2 group, wherein the NH.sub.2 side chain group is optionally functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms selected from halogen, N, O, S, P, and combinations thereof, R.sup.1 is the N-terminal group of the peptidic compound and is selected from NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.5)alkyl], N[(C.sub.1-C.sub.5)alkyl].sub.2, NH[(C.sub.0-C.sub.4)alkylene-(C.sub.3-C.sub.8)cycloalkyl], NHC(O)H, NHC(O)(C.sub.1-C.sub.5)-alkyl, and NHC(O)(C.sub.0-C.sub.3)alkylene-(C.sub.3-C.sub.8)cycloalkyl, in which alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or halogen selected from F, Cl, Br, and I, R.sup.2 is the C-terminal group of the peptidic compound and is selected from (i) OH and functionalized OH, wherein the functionalized OH is selected from O(C.sub.1-C.sub.20)alkyl and O(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl, and (ii) NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.30)alkyl], N[(C.sub.1-C.sub.30)alkyl].sub.2, NH[(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl], N[C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl].sub.2, NH[(CH.sub.2CH.sub.2O).sub.1-40(C.sub.1-C.sub.4)alkyl], NH(C.sub.3-C.sub.8)heterocyclyl or NH(C.sub.0-C.sub.8)alkylene-aryl, wherein aryl is selected from phenyl or naphthyl, the (C.sub.3-C.sub.8)-heterocyclyl contains one N-atom and optionally two additional heteroatoms selected from O, N and S, and alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or a halogen selected from F, Cl, Br, and I.
35. The method according to claim 34, wherein said peptidic compound of formula (I) is any one of SEQ ID NOs: 4-181, 196-223, 226-229, or a salt or solvate thereof.
36. The method according to claim 34, wherein said method comprises treating one or more vascular disease conditions in the patient, wherein the one or more vascular disease conditions are selected from the group consisting of ischemic heart disease, myocardial infarction, stroke, peripheral vascular disease, diabetic retinopathy, diabetic nephropathy, and diabetic neuropathy.
37. A method for delaying disease progression of type 2 diabetes in a patient in need thereof, the method comprising administering to said patient a therapeutically effective amount of the peptidic compound of formula (I):
R.sup.1ZR.sup.2(I) or a salt, or solvate thereof, or a pharmaceutical composition comprising a compound of formula (I) or a salt or solvate thereof as an active agent together with at least one pharmaceutically acceptable carrier, wherein Z is a peptide moiety having formula (II): TABLE-US-00022 (II) His-X2-X3-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln- X14-X15-X16-X17-X18-Ala-X20-X21-Phe-Ile-Glu-Trp- Leu-Lys-X28-X29-Gly-Pro-Ser-Ser-Gly-X35-Pro-Pro- Pro-X39-X40 wherein: X2 is an amino acid residue selected from Ser, D-Ser, and Aib, X3 is an amino acid residue selected from Gln, His, and -amino-functionalized Gln, wherein Gln is optionally functionalized in that an H of the -NH.sub.2 group is substituted by (C.sub.1-C.sub.4)-alkyl, X14 is an amino acid residue having a side chain with a functionalized NH.sub.2 group, wherein the functionalized NH.sub.2 side chain group is functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms independently selected from halogen, N, O, S, P, and combinations thereof, X15 is an amino acid residue selected from Glu and Asp, X16 is an amino acid residue selected from Ser, Glu, and Lys, X17 is an amino acid residue selected from Arg, Glu, Gln, Leu, Aib, and Lys, X18 is an amino acid residue selected from Arg, Ala, and Lys, X20 is an amino acid residue selected from Gln, Arg, Lys, His, Glu, and Aib, X21 is an amino acid residue selected from Asp, Leu, and Glu, X28 is an amino acid residue selected from Asn, Arg, Lys, Aib, Ser, Glu, Ala, and Asp, X29 is an amino acid residue selected from Gly, Ala, D-Ala, and Thr, X35 is an amino acid residue selected from Ala, Glu, Arg, and Lys, X39 is Ser or is absent, and X40 is absent or is an amino acid residue having a side chain with an NH.sub.2 group, wherein the NH.sub.2 side chain group is optionally functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms selected from halogen, N, O, S, P, and combinations thereof, R.sup.1 is the N-terminal group of the peptidic compound and is selected from NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.5)alkyl], N[(C.sub.1-C.sub.5)alkyl].sub.2, NH[(C.sub.0-C.sub.4)alkylene-(C.sub.3-C.sub.8)cycloalkyl], NHC(O)H, NHC(O)(C.sub.1-C.sub.5)-alkyl, and NHC(O)(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl, in which alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or halogen selected from F, Cl, Br, and I, R.sup.2 is the C-terminal group of the peptidic compound and is selected from (i) OH and functionalized OH, wherein the functionalized OH is selected from O(C.sub.1-C.sub.20)alkyl and O(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl, and (ii) NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.30)alkyl], N[(C.sub.1-C.sub.30)alkyl].sub.2, NH[(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl], N[C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl].sub.2, NH[(CH.sub.2CH.sub.2O).sub.1-40(C.sub.1-C.sub.4)alkyl], NH(C.sub.3-C.sub.8)heterocyclyl or NH(C.sub.0-C.sub.8)alkylene-aryl, wherein aryl is selected from phenyl or naphthyl, the (C.sub.3-C.sub.8)-heterocyclyl contains one N-atom and optionally two additional heteroatoms selected from O, N and S, and alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or a halogen selected from F, Cl, Br, and I.
38. The method according to claim 37, wherein said peptidic compound of formula (I) is any one of SEQ ID NOs: 4-181, 196-223, 226-229, or a salt or solvate thereof.
39. The method of claim 37, wherein said method comprises delaying the progression from type 2 diabetes to insulin-requiring diabetes.
40. A method for treating impaired glucose tolerance in a patient in need thereof, the method comprising administering to said patient a therapeutically effective amount of the peptidic compound of formula (I):
R.sup.1ZR.sup.2(I) or a salt, or solvate thereof, or a pharmaceutical composition comprising a compound of formula (I) or a salt or solvate thereof as an active agent together with at least one pharmaceutically acceptable carrier, wherein Z is a peptide moiety having formula (II): TABLE-US-00023 (II) His-X2-X3-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln- X14-X15-X16-X17-X18-Ala-X20-X21-Phe-Ile-Glu-Trp- Leu-Lys-X28-X29-Gly-Pro-Ser-Ser-Gly-X35-Pro-Pro- Pro-X39-X40 wherein: X2 is an amino acid residue selected from Ser, D-Ser, and Aib, X3 is an amino acid residue selected from Gln, His, and -amino-functionalized Gln, wherein Gln is optionally functionalized in that an H of the -NH.sub.2 group is substituted by (C.sub.1-C.sub.4)-alkyl, X14 is an amino acid residue having a side chain with a functionalized NH.sub.2 group, wherein the functionalized NH.sub.2 side chain group is functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms independently selected from halogen, N, O, S, P, and combinations thereof, X15 is an amino acid residue selected from Glu and Asp, X16 is an amino acid residue selected from Ser, Glu, and Lys, X17 is an amino acid residue selected from Arg, Glu, Gln, Leu, Aib, and Lys, X18 is an amino acid residue selected from Arg, Ala, and Lys, X20 is an amino acid residue selected from Gln, Arg, Lys, His, Glu, and Aib, X21 is an amino acid residue selected from Asp, Leu, and Glu, X28 is an amino acid residue selected from Asn, Arg, Lys, Aib, Ser, Glu, Ala, and Asp, X29 is an amino acid residue selected from Gly, Ala, D-Ala, and Thr, X35 is an amino acid residue selected from Ala, Glu, Arg, and Lys, X39 is Ser or is absent, and X40 is absent or is an amino acid residue having a side chain with an NH.sub.2 group, wherein the NH.sub.2 side chain group is optionally functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms selected from halogen, N, O, S, P, and combinations thereof, R.sup.1 is the N-terminal group of the peptidic compound and is selected from NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.5)alkyl], N[(C.sub.1-C.sub.5)alkyl].sub.2, NH[(C.sub.0-C.sub.4)alkylene-(C.sub.3-C.sub.8)cycloalkyl], NHC(O)H, NHC(O)(C.sub.1-C.sub.5)-alkyl, and NHC(O)(C.sub.0-C.sub.3)alkylene-(C.sub.3-C.sub.8)cycloalkyl, in which alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or halogen selected from F, Cl, Br, and I, R.sup.2 is the C-terminal group of the peptidic compound and is selected from (i) OH and functionalized OH, wherein the functionalized OH is selected from O(C.sub.1-C.sub.20)alkyl and O(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl, and (ii) NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.30)alkyl], N[(C.sub.1-C.sub.30)alkyl].sub.2, NH[(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl], N[C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl].sub.2, NH[(CH.sub.2CH.sub.2O).sub.1-40(C.sub.1-C.sub.4)alkyl], NH(C.sub.3-C.sub.8)heterocyclyl or NH(C.sub.0-C.sub.8)alkylene-aryl, wherein aryl is selected from phenyl or naphthyl, the (C.sub.3-C.sub.8)-heterocyclyl contains one N-atom and optionally two additional heteroatoms selected from O, N and S, and alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or a halogen selected from F, Cl, Br, and I.
41. The method according to claim 40, wherein said peptidic compound of formula (I) is any one of SEQ ID NOs: 4-181, 196-223, 226-229, or a salt or solvate thereof.
42. A method for reducing a blood glucose level in a patient in need thereof, the method comprising administering to said patient a therapeutically effective amount of the peptidic compound of formula (I):
R.sup.1ZR.sup.2(I) or a salt, or solvate thereof, or a pharmaceutical composition comprising a compound of formula (I) or a salt or solvate thereof as an active agent together with at least one pharmaceutically acceptable carrier, wherein Z is a peptide moiety having formula (II): TABLE-US-00024 (II) His-X2-X3-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln- X14-X15-X16-X17-X18-Ala-X20-X21-Phe-Ile-Glu-Trp- Leu-Lys-X28-X29-Gly-Pro-Ser-Ser-Gly-X35-Pro-Pro- Pro-X39-X40 wherein: X2 is an amino acid residue selected from Ser, D-Ser, and Aib, X3 is an amino acid residue selected from Gln, His, and -amino-functionalized Gln, wherein Gln is optionally functionalized in that an H of the -NH.sub.2 group is substituted by (C.sub.1-C.sub.4)-alkyl, X14 is an amino acid residue having a side chain with a functionalized NH.sub.2 group, wherein the functionalized NH.sub.2 side chain group is functionalized by C(O)-R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms independently selected from halogen, N, O, S, P, and combinations thereof, X15 is an amino acid residue selected from Glu and Asp, X16 is an amino acid residue selected from Ser, Glu, and Lys, X17 is an amino acid residue selected from Arg, Glu, Gln, Leu, Aib, and Lys, X18 is an amino acid residue selected from Arg, Ala, and Lys, X20 is an amino acid residue selected from Gln, Arg, Lys, His, Glu, and Aib, X21 is an amino acid residue selected from Asp, Leu, and Glu, X28 is an amino acid residue selected from Asn, Arg, Lys, Aib, Ser, Glu, Ala, and Asp, X29 is an amino acid residue selected from Gly, Ala, D-Ala, and Thr, X35 is an amino acid residue selected from Ala, Glu, Arg, and Lys, X39 is Ser or is absent, and X40 is absent or is an amino acid residue having a side chain with an NH.sub.2 group, wherein the NH.sub.2 side chain group is optionally functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms selected from halogen, N, O, S, P, and combinations thereof, R.sup.1 is the N-terminal group of the peptidic compound and is selected from NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.5)alkyl], N[(C.sub.1-C.sub.5)alkyl].sub.2, NH[(C.sub.0-C.sub.4)alkylene-(C.sub.3-C.sub.8)cycloalkyl], NHC(O)H, NHC(O)(C.sub.1-C.sub.5)-alkyl, and NHC(O)(C.sub.0-C.sub.3)alkylene-(C.sub.3-C.sub.8)cycloalkyl, in which alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or halogen selected from F, Cl, Br, and I, R.sup.2 is the C-terminal group of the peptidic compound and is selected from (i) OH and functionalized OH, wherein the functionalized OH is selected from O(C.sub.1-C.sub.20)alkyl and O(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl, and (ii) NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.30)alkyl], N[(C.sub.1-C.sub.30)alkyl].sub.2, NH[(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl], N[C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl].sub.2, NH[(CH.sub.2CH.sub.2O).sub.1-40(C.sub.1-C.sub.4)alkyl], NH(C.sub.3-C.sub.8)heterocyclyl or NH(C.sub.0-C.sub.8)alkylene-aryl, wherein aryl is selected from phenyl or naphthyl, the (C.sub.3-C.sub.8)-heterocyclyl contains one N-atom and optionally two additional heteroatoms selected from O, N and S, and alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or a halogen selected from F, Cl, Br, and I.
43. The method according to claim 42, wherein said peptidic compound of formula (I) is any one of SEQ ID NOs: 4-181, 196-223, 226-229, or a salt or solvate thereof.
44. A method for reducing an HbA1c level in a patient in need thereof, the method comprising administering to said patient a therapeutically effective amount of the peptidic compound of formula (I):
R.sup.1ZR.sup.2(I) or a salt, or solvate thereof, or a pharmaceutical composition comprising a compound of formula (I) or a salt or solvate thereof as an active agent together with at least one pharmaceutically acceptable carrier, wherein Z is a peptide moiety having formula (II): TABLE-US-00025 (II) His-X2-X3-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln- X14-X15-X16-X17-X18-Ala-X20-X21-Phe-Ile-Glu-Trp- Leu-Lys-X28-X29-Gly-Pro-Ser-Ser-Gly-X35-Pro-Pro- Pro-X39-X40 wherein: X2 is an amino acid residue selected from Ser, D-Ser, and Aib, X3 is an amino acid residue selected from Gln, His, and -amino-functionalized Gln, wherein Gln is optionally functionalized in that an H of the -NH.sub.2 group is substituted by (C.sub.1-C.sub.4)-alkyl, X14 is an amino acid residue having a side chain with a functionalized NH.sub.2 group, wherein the functionalized NH.sub.2 side chain group is functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms independently selected from halogen, N, O, S, P, and combinations thereof, X15 is an amino acid residue selected from Glu and Asp, X16 is an amino acid residue selected from Ser, Glu, and Lys, X17 is an amino acid residue selected from Arg, Glu, Gln, Leu, Aib, and Lys, X18 is an amino acid residue selected from Arg, Ala, and Lys, X20 is an amino acid residue selected from Gln, Arg, Lys, His, Glu, and Aib, X21 is an amino acid residue selected from Asp, Leu, and Glu, X28 is an amino acid residue selected from Asn, Arg, Lys, Aib, Ser, Glu, Ala, and Asp, X29 is an amino acid residue selected from Gly, Ala, D-Ala, and Thr, X35 is an amino acid residue selected from Ala, Glu, Arg, and Lys, X39 is Ser or is absent, and X40 is absent or is an amino acid residue having a side chain with an NH.sub.2 group, wherein the NH.sub.2 side chain group is optionally functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms selected from halogen, N, O, S, P, and combinations thereof, R.sup.1 is the N-terminal group of the peptidic compound and is selected from NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.5)alkyl], N[(C.sub.1-C.sub.5)alkyl].sub.2, NH[(C.sub.0-C.sub.4)alkylene-(C.sub.3-C.sub.8)cycloalkyl], NHC(O)H, NHC(O)(C.sub.1-C.sub.5)-alkyl, and NHC(O)(C.sub.0-C.sub.3)alkylene-(C.sub.3-C.sub.8)cycloalkyl, in which alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or halogen selected from F, Cl, Br, and I, R.sup.2 is the C-terminal group of the peptidic compound and is selected from (i) OH and functionalized OH, wherein the functionalized OH is selected from O(C.sub.1-C.sub.20)alkyl and O(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl, and (ii) NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.30)alkyl], N[(C.sub.1-C.sub.30)alkyl].sub.2, NH[(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl], N[C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl].sub.2, NH[(CH.sub.2CH.sub.2O).sub.1-40(C.sub.1-C.sub.4)alkyl], NH(C.sub.3-C.sub.8)heterocyclyl or NH(C.sub.0-C.sub.8)alkylene-aryl, wherein aryl is selected from phenyl or naphthyl, the (C.sub.3-C.sub.8)-heterocyclyl contains one N-atom and optionally two additional heteroatoms selected from O, N and S, and alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or a halogen selected from F, Cl, Br, and I.
45. The method according to claim 44, wherein said peptidic compound of formula (I) is any one of SEQ ID NOs: 4-181, 196-223, 226-229, or a salt or solvate thereof.
46. A method for delaying disease progression from impaired glucose tolerance (IGT) to type 2 diabetes in a patient in need thereof, the method comprising administering to said patient a therapeutically effective amount of the peptidic compound of formula (I):
R.sup.1ZR.sup.2(I) or a salt, or solvate thereof, or a pharmaceutical composition comprising a compound of formula (I) or a salt or solvate thereof as an active agent together with at least one pharmaceutically acceptable carrier, wherein Z is a peptide moiety having formula (II): TABLE-US-00026 (II) His-X2-X3-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln- X14-X15-X16-X17-X18-Ala-X20-X21-Phe-Ile-Glu-Trp- Leu-Lys-X28-X29-Gly-Pro-Ser-Ser-Gly-X35-Pro-Pro- Pro-X39-X40 wherein: X2 is an amino acid residue selected from Ser, D-Ser, and Aib, X3 is an amino acid residue selected from Gln, His, and -amino-functionalized Gln, wherein Gln is optionally functionalized in that an H of the -NH.sub.2 group is substituted by (C.sub.1-C.sub.4)-alkyl, X14 is an amino acid residue having a side chain with a functionalized NH.sub.2 group, wherein the functionalized NH.sub.2 side chain group is functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms independently selected from halogen, N, O, S, P, and combinations thereof, X15 is an amino acid residue selected from Glu and Asp, X16 is an amino acid residue selected from Ser, Glu, and Lys, X17 is an amino acid residue selected from Arg, Glu, Gln, Leu, Aib, and Lys, X18 is an amino acid residue selected from Arg, Ala, and Lys, X20 is an amino acid residue selected from Gln, Arg, Lys, His, Glu, and Aib, X21 is an amino acid residue selected from Asp, Leu, and Glu, X28 is an amino acid residue selected from Asn, Arg, Lys, Aib, Ser, Glu, Ala, and Asp, X29 is an amino acid residue selected from Gly, Ala, D-Ala, and Thr, X35 is an amino acid residue selected from Ala, Glu, Arg, and Lys, X39 is Ser or is absent, and X40 is absent or is an amino acid residue having a side chain with an NH.sub.2 group, wherein the NH.sub.2 side chain group is optionally functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms selected from halogen, N, O, S, P, and combinations thereof, R.sup.1 is the N-terminal group of the peptidic compound and is selected from NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.5)alkyl], N[(C.sub.1-C.sub.5)alkyl].sub.2, NH[(C.sub.0-C.sub.4)alkylene-(C.sub.3-C.sub.8)cycloalkyl], NHC(O)H, NHC(O)(C.sub.1-C.sub.5)-alkyl, and NHC(O)(C.sub.0-C.sub.3)alkylene-(C.sub.3-C.sub.8)cycloalkyl, in which alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or halogen selected from F, Cl, Br, and I, R.sup.2 is the C-terminal group of the peptidic compound and is selected from (i) OH and functionalized OH, wherein the functionalized OH is selected from O(C.sub.1-C.sub.20)alkyl and O(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl, and (ii) NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.30)alkyl], N[(C.sub.1-C.sub.30)alkyl].sub.2, NH[(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl], N[C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl].sub.2, NH[(CH.sub.2CH.sub.2O).sub.1-40(C.sub.1-C.sub.4)alkyl], NH(C.sub.3-C.sub.8)heterocyclyl or NH(C.sub.0-C.sub.8)alkylene-aryl, wherein aryl is selected from phenyl or naphthyl, the (C.sub.3-C.sub.8)-heterocyclyl contains one N-atom and optionally two additional heteroatoms selected from O, N and S, and alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or a halogen selected from F, Cl, Br, and I.
47. The method according to claim 46, wherein said peptidic compound of formula (I) is any one of SEQ ID NOs: 4-181, 196-223, 226-229, or a salt or solvate thereof.
48. A method for treating hyperglycemia in a patient in need thereof, the method comprising administering to said patient a therapeutically effective amount of the peptidic compound of formula (I):
R.sup.1ZR.sup.2(I) or a salt, or solvate thereof, or a pharmaceutical composition comprising a compound of formula (I) or a salt or solvate thereof as an active agent together with at least one pharmaceutically acceptable carrier, wherein Z is a peptide moiety having formula (II): TABLE-US-00027 (II) His-X2-X3-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln- X14-X15-X16-X17-X18-Ala-X20-X21-Phe-Ile-Glu-Trp- Leu-Lys-X28-X29-Gly-Pro-Ser-Ser-Gly-X35-Pro-Pro- Pro-X39-X40 wherein: X2 is an amino acid residue selected from Ser, D-Ser, and Aib, X3 is an amino acid residue selected from Gln, His, and -amino-functionalized Gln, wherein Gln is optionally functionalized in that an H of the -NH.sub.2 group is substituted by (C.sub.1-C.sub.4)-alkyl, X14 is an amino acid residue having a side chain with a functionalized NH.sub.2 group, wherein the functionalized NH.sub.2 side chain group is functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms independently selected from halogen, N, O, S, P, and combinations thereof, X15 is an amino acid residue selected from Glu and Asp, X16 is an amino acid residue selected from Ser, Glu, and Lys, X17 is an amino acid residue selected from Arg, Glu, Gln, Leu, Aib, and Lys, X18 is an amino acid residue selected from Arg, Ala, and Lys, X20 is an amino acid residue selected from Gln, Arg, Lys, His, Glu, and Aib, X21 is an amino acid residue selected from Asp, Leu, and Glu, X28 is an amino acid residue selected from Asn, Arg, Lys, Aib, Ser, Glu, Ala, and Asp, X29 is an amino acid residue selected from Gly, Ala, D-Ala, and Thr, X35 is an amino acid residue selected from Ala, Glu, Arg, and Lys, X39 is Ser or is absent, and X40 is absent or is an amino acid residue having a side chain with an NH.sub.2 group, wherein the NH.sub.2 side chain group is optionally functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms selected from halogen, N, O, S, P, and combinations thereof, R.sup.1 is the N-terminal group of the peptidic compound and is selected from NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.5)alkyl], N[(C.sub.1-C.sub.5)alkyl].sub.2, NH[(C.sub.0-C.sub.4)alkylene-(C.sub.3-C.sub.8)cycloalkyl], NHC(O)H, NHC(O)(C.sub.1-C.sub.5)-alkyl, and NHC(O)(C.sub.0-C.sub.3)alkylene-(C.sub.3-C.sub.8)cycloalkyl, in which alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or halogen selected from F, Cl, Br, and I, R.sup.2 is the C-terminal group of the peptidic compound and is selected from (i) OH and functionalized OH, wherein the functionalized OH is selected from O(C.sub.1-C.sub.20)alkyl and O(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl, and (ii) NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.30)alkyl], N[(C.sub.1-C.sub.30)alkyl].sub.2, NH[(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl], N[C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl].sub.2, NH[(CH.sub.2CH.sub.2O).sub.1-40(C.sub.1-C.sub.4)alkyl], NH(C.sub.3-C.sub.8)heterocyclyl or NH(C.sub.0-C.sub.8)alkylene-aryl, wherein aryl is selected from phenyl or naphthyl, the (C.sub.3-C.sub.8)-heterocyclyl contains one N-atom and optionally two additional heteroatoms selected from O, N and S, and alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or a halogen selected from F, Cl, Br, and I.
49. The method according to claim 48, wherein said peptidic compound of formula (I) is any one of SEQ ID NOs: 4-181, 196-223, 226-229, or a salt or solvate thereof.
50. A method for treating obesity in a patient in need thereof, the method comprising administering to said patient a therapeutically effective amount of the peptidic compound of formula (I):
R.sup.1ZR.sup.2(I) or a salt, or solvate thereof, or a pharmaceutical composition comprising a compound of formula (I) or a salt or solvate thereof as an active agent together with at least one pharmaceutically acceptable carrier, wherein Z is a peptide moiety having formula (II): TABLE-US-00028 (II) His-X2-X3-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln- X14-X15-X16-X17-X18-Ala-X20-X21-Phe-Ile-Glu-Trp- Leu-Lys-X28-X29-Gly-Pro-Ser-Ser-Gly-X35-Pro-Pro- Pro-X39-X40 wherein: X2 is an amino acid residue selected from Ser, D-Ser, and Aib, X3 is an amino acid residue selected from Gln, His, and -amino-functionalized Gln, wherein Gln is optionally functionalized in that an H of the -NH.sub.2 group is substituted by (C.sub.1-C.sub.4)-alkyl, X14 is an amino acid residue having a side chain with a functionalized NH.sub.2 group, wherein the functionalized NH.sub.2 side chain group is functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms independently selected from halogen, N, O, S, P, and combinations thereof, X15 is an amino acid residue selected from Glu and Asp, X16 is an amino acid residue selected from Ser, Glu, and Lys, X17 is an amino acid residue selected from Arg, Glu, Gln, Leu, Aib, and Lys, X18 is an amino acid residue selected from Arg, Ala, and Lys, X20 is an amino acid residue selected from Gln, Arg, Lys, His, Glu, and Aib, X21 is an amino acid residue selected from Asp, Leu, and Glu, X28 is an amino acid residue selected from Asn, Arg, Lys, Aib, Ser, Glu, Ala, and Asp, X29 is an amino acid residue selected from Gly, Ala, D-Ala, and Thr, X35 is an amino acid residue selected from Ala, Glu, Arg, and Lys, X39 is Ser or is absent, and X40 is absent or is an amino acid residue having a side chain with an NH.sub.2 group, wherein the NH.sub.2 side chain group is optionally functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms selected from halogen, N, O, S, P, and combinations thereof, R.sup.1 is the N-terminal group of the peptidic compound and is selected from NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.5)alkyl], N[(C.sub.1-C.sub.5)alkyl].sub.2, NH[(C.sub.0-C.sub.4)alkylene-(C.sub.3-C.sub.8)cycloalkyl], NHC(O)H, NHC(O)(C.sub.1-C.sub.5)-alkyl, and NHC(O)(C.sub.0-C.sub.3)alkylene-(C.sub.3-C.sub.8)cycloalkyl, in which alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or halogen selected from F, Cl, Br, and I, R.sup.2 is the C-terminal group of the peptidic compound and is selected from (i) OH and functionalized OH, wherein the functionalized OH is selected from O(C.sub.1-C.sub.20)alkyl and O(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl, and (ii) NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.30)alkyl], N[(C.sub.1-C.sub.30)alkyl].sub.2, NH[(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl], N[C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl].sub.2, NH[(CH.sub.2CH.sub.2O).sub.140(C.sub.1-C.sub.4)alkyl], NH(C.sub.3-C.sub.8)heterocyclyl or NH(C.sub.0-C.sub.8)alkylene-aryl, wherein aryl is selected from phenyl or naphthyl, the (C.sub.3-C.sub.8)-heterocyclyl contains one N-atom and optionally two additional heteroatoms selected from O, N and S, and alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or a halogen selected from F, Cl, Br, and I.
51. The method according to claim 50, wherein said peptidic compound of formula (I) is any one of SEQ ID NOs: 4-181, 196-223, 226-229, or a salt or solvate thereof.
52. The method according to claim 50, further comprising treating obesity-linked inflammation, obesity-linked gallbladder disease, and/or obesity induced sleep apnea in the patient.
53. A method for decreasing food intake, reducing body weight, regulating appetite, or inducing satiety in a patient in need thereof, the method comprising administering to said patient a therapeutically effective amount of the peptidic compound of formula (I):
R.sup.1ZR.sup.2(I) or a salt, or solvate thereof, or a pharmaceutical composition comprising a compound of formula (I) or a salt or solvate thereof as an active agent together with at least one pharmaceutically acceptable carrier, wherein Z is a peptide moiety having formula (II): TABLE-US-00029 (II) His-X2-X3-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln- X14-X15-X16-X17-X18-Ala-X20-X21-Phe-Ile-Glu-Trp- Leu-Lys-X28-X29-Gly-Pro-Ser-Ser-Gly-X35-Pro-Pro- Pro-X39-X40 wherein: X2 is an amino acid residue selected from Ser, D-Ser, and Aib, X3 is an amino acid residue selected from Gln, His, and -amino-functionalized Gln, wherein Gln is optionally functionalized in that an H of the -NH.sub.2 group is substituted by (C.sub.1-C.sub.4)-alkyl, X14 is an amino acid residue having a side chain with a functionalized NH.sub.2 group, wherein the functionalized NH.sub.2 side chain group is functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms independently selected from halogen, N, O, S, P, and combinations thereof, X15 is an amino acid residue selected from Glu and Asp, X16 is an amino acid residue selected from Ser, Glu, and Lys, X17 is an amino acid residue selected from Arg, Glu, Gln, Leu, Aib, and Lys, X18 is an amino acid residue selected from Arg, Ala, and Lys, X20 is an amino acid residue selected from Gln, Arg, Lys, His, Glu, and Aib, X21 is an amino acid residue selected from Asp, Leu, and Glu, X28 is an amino acid residue selected from Asn, Arg, Lys, Aib, Ser, Glu, Ala, and Asp, X29 is an amino acid residue selected from Gly, Ala, D-Ala, and Thr, X35 is an amino acid residue selected from Ala, Glu, Arg, and Lys, X39 is Ser or is absent, and X40 is absent or is an amino acid residue having a side chain with an NH.sub.2 group, wherein the NH.sub.2 side chain group is optionally functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms selected from halogen, N, O, S, P, and combinations thereof, R.sup.1 is the N-terminal group of the peptidic compound and is selected from NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.5)alkyl], N[(C.sub.1-C.sub.5)alkyl].sub.2, NH[(C.sub.0-C.sub.4)alkylene-(C.sub.3-C.sub.8)cycloalkyl], NHC(O)H, NHC(O)(C.sub.1-C.sub.5)-alkyl, and NHC(O)(C.sub.0-C.sub.3)alkylene-(C.sub.3-C.sub.8)cycloalkyl, in which alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or halogen selected from F, Cl, Br, and I, R.sup.2 is the C-terminal group of the peptidic compound and is selected from (i) OH and functionalized OH, wherein the functionalized OH is selected from O(C.sub.1-C.sub.20)alkyl and O(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl, and (ii) NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.30)alkyl], N[(C.sub.1-C.sub.30)alkyl].sub.2, NH[(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl], N[C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl].sub.2, NH[(CH.sub.2CH.sub.2O).sub.1-40(C.sub.1-C.sub.4)alkyl], NH(C.sub.3-C.sub.8)heterocyclyl or NH(C.sub.0-C.sub.8)alkylene-aryl, wherein aryl is selected from phenyl or naphthyl, the (C.sub.3-C.sub.8)-heterocyclyl contains one N-atom and optionally two additional heteroatoms selected from O, N and S, and alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or a halogen selected from F, Cl, Br, and I.
54. The method according to claim 53, wherein said peptidic compound of formula (I) is any one of SEQ ID NOs: 4-181, 196-223, 226-229, or a salt or solvate thereof.
55. A method for inhibiting the motility of the gastrointestinal tract in a patient in need thereof, the method comprising administering to said patient a therapeutically effective amount of the peptidic compound of formula (I):
R.sup.1ZR.sup.2(I) or a salt, or solvate thereof, or a pharmaceutical composition comprising a compound of formula (I) or a salt or solvate thereof as an active agent together with at least one pharmaceutically acceptable carrier, wherein Z is a peptide moiety having formula (II): TABLE-US-00030 (II) His-X2-X3-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln- X14-X15-X16-X17-X18-Ala-X20-X21-Phe-Ile-Glu-Trp- Leu-Lys-X28-X29-Gly-Pro-Ser-Ser-Gly-X35-Pro-Pro- Pro-X39-X40 wherein: X2 is an amino acid residue selected from Ser, D-Ser, and Aib, X3 is an amino acid residue selected from Gln, His, and -amino-functionalized Gln, wherein Gln is optionally functionalized in that an H of the -NH.sub.2 group is substituted by (C.sub.1-C.sub.4)-alkyl, X14 is an amino acid residue having a side chain with a functionalized NH.sub.2 group, wherein the functionalized NH.sub.2 side chain group is functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms independently selected from halogen, N, O, S, P, and combinations thereof, X15 is an amino acid residue selected from Glu and Asp, X16 is an amino acid residue selected from Ser, Glu, and Lys, X17 is an amino acid residue selected from Arg, Glu, Gln, Leu, Aib, and Lys, X18 is an amino acid residue selected from Arg, Ala, and Lys, X20 is an amino acid residue selected from Gln, Arg, Lys, His, Glu, and Aib, X21 is an amino acid residue selected from Asp, Leu, and Glu, X28 is an amino acid residue selected from Asn, Arg, Lys, Aib, Ser, Glu, Ala, and Asp, X29 is an amino acid residue selected from Gly, Ala, D-Ala, and Thr, X35 is an amino acid residue selected from Ala, Glu, Arg, and Lys, X39 is Ser or is absent, and X40 is absent or is an amino acid residue having a side chain with an NH.sub.2 group, wherein the NH.sub.2 side chain group is optionally functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms selected from halogen, N, O, S, P, and combinations thereof, R.sup.1 is the N-terminal group of the peptidic compound and is selected from NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.5)alkyl], N[(C.sub.1-C.sub.5)alkyl].sub.2, NH[(C.sub.0-C.sub.4)alkylene-(C.sub.3-C.sub.8)cycloalkyl], NHC(O)H, NHC(O)(C.sub.1-C.sub.5)-alkyl, and NHC(O)(C.sub.0-C.sub.3)alkylene-(C.sub.3-C.sub.8)cycloalkyl, in which alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or halogen selected from F, Cl, Br, and I, R.sup.2 is the C-terminal group of the peptidic compound and is selected from (i) OH and functionalized OH, wherein the functionalized OH is selected from O(C.sub.1-C.sub.20)alkyl and O(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl, and (ii) NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.30)alkyl], N[(C.sub.1-C.sub.30)alkyl].sub.2, NH[(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl], N[C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl].sub.2, NH[(CH.sub.2CH.sub.2O).sub.1-40(C.sub.1-C.sub.4)alkyl], NH(C.sub.3-C.sub.8)heterocyclyl or NH(C.sub.0-C.sub.8)alkylene-aryl, wherein aryl is selected from phenyl or naphthyl, the (C.sub.3-C.sub.8)-heterocyclyl contains one N-atom and optionally two additional heteroatoms selected from O, N and S, and alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or a halogen selected from F, Cl, Br, and I.
56. The method according to claim 55, wherein said peptidic compound of formula (I) is any one of SEQ ID NOs: 4-181, 196-223, 226-229, or a salt or solvate thereof.
57. A method for treating a neurodegenerative disorder in a patient in need thereof, the method comprising administering to said patient a therapeutically effective amount of the peptidic compound of formula (I):
R.sup.1ZR.sup.2(I) or a salt, or solvate thereof, or a pharmaceutical composition comprising a compound of formula (I) or a salt or solvate thereof as an active agent together with at least one pharmaceutically acceptable carrier, wherein Z is a peptide moiety having formula (II): TABLE-US-00031 (II) His-X2-X3-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln- X14-X15-X16-X17-X18-Ala-X20-X21-Phe-Ile-Glu-Trp- Leu-Lys-X28-X29-Gly-Pro-Ser-Ser-Gly-X35-Pro-Pro- Pro-X39-X40 wherein: X2 is an amino acid residue selected from Ser, D-Ser, and Aib, X3 is an amino acid residue selected from Gln, His, and -amino-functionalized Gln, wherein Gln is optionally functionalized in that an H of the -NH.sub.2 group is substituted by (C.sub.1-C.sub.4)-alkyl, X14 is an amino acid residue having a side chain with a functionalized NH.sub.2 group, wherein the functionalized NH.sub.2 side chain group is functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms independently selected from halogen, N, O, S, P, and combinations thereof, X15 is an amino acid residue selected from Glu and Asp, X16 is an amino acid residue selected from Ser, Glu, and Lys, X17 is an amino acid residue selected from Arg, Glu, Gln, Leu, Aib, and Lys, X18 is an amino acid residue selected from Arg, Ala, and Lys, X20 is an amino acid residue selected from Gln, Arg, Lys, His, Glu, and Aib, X21 is an amino acid residue selected from Asp, Leu, and Glu, X28 is an amino acid residue selected from Asn, Arg, Lys, Aib, Ser, Glu, Ala, and Asp, X29 is an amino acid residue selected from Gly, Ala, D-Ala, and Thr, X35 is an amino acid residue selected from Ala, Glu, Arg, and Lys, X39 is Ser or is absent, and X40 is absent or is an amino acid residue having a side chain with an NH.sub.2 group, wherein the NH.sub.2 side chain group is optionally functionalized by C(O)R.sup.5, C(O)OR.sup.5, C(O)NHR.sup.5, S(O).sub.2R.sup.5 or R.sup.5, wherein R.sup.5 is a moiety comprising up to 100 carbon atoms and optionally heteroatoms selected from halogen, N, O, S, P, and combinations thereof, R.sup.1 is the N-terminal group of the peptidic compound and is selected from NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.5)alkyl], N[(C.sub.1-C.sub.5)alkyl].sub.2, NH[(C.sub.0-C.sub.4)alkylene-(C.sub.3-C.sub.8)cycloalkyl], NHC(O)H, NHC(O)(C.sub.1-C.sub.5)-alkyl, and NHC(O)(C.sub.0-C.sub.3)alkylene-(C.sub.3-C.sub.8)cycloalkyl, in which alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or halogen selected from F, Cl, Br, and I, R.sup.2 is the C-terminal group of the peptidic compound and is selected from (i) OH and functionalized OH, wherein the functionalized OH is selected from O(C.sub.1-C.sub.20)alkyl and O(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl, and (ii) NH.sub.2 and mono- or bisfunctionalized NH.sub.2, wherein the mono- or bisfunctionalized NH.sub.2 is selected from the group consisting of NH[(C.sub.1-C.sub.30)alkyl], N[(C.sub.1-C.sub.30)alkyl].sub.2, NH[(C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl], N[C.sub.0-C.sub.8)alkylene-(C.sub.3-C.sub.8)cycloalkyl].sub.2, NH[(CH.sub.2CH.sub.2O).sub.1-40(C.sub.1-C.sub.4)alkyl], NH(C.sub.3-C.sub.8)heterocyclyl or NH(C.sub.0-C.sub.8)alkylene-aryl, wherein aryl is selected from phenyl or naphthyl, the (C.sub.3-C.sub.8)-heterocyclyl contains one N-atom and optionally two additional heteroatoms selected from O, N and S, and alkyl or cycloalkyl is unsubstituted or up to 5-fold substituted by OH or a halogen selected from F, Cl, Br, and I.
58. The method of claim 57, wherein the neurodegenerative disorder is selected from the group consisting of Alzheimer's disease, Parkinson's disease, Huntington's disease, ataxia, Kennedy disease, myotonic dystrophy, Lewy body dementia, multi-systemic atrophy, amyotrophic lateral sclerosis, primary lateral sclerosis, spinal muscular atrophy, prion-associated diseases, multiple sclerosis, telangiectasia, Batten disease, corticobasal degeneration, subacute combined degeneration of spinal cord, Tabes dorsalis, Tay-Sachs disease, toxic encephalopathy, infantile Refsum disease, Refsum disease, neuroacanthocytosis, Niemann-Pick disease, Lyme disease, Machado-Joseph disease, Sandhoff disease, Shy-Drager syndrome, wobbly hedgehog syndrome, proteopathy, cerebral p-amyloid angiopathy, retinal ganglion cell degeneration in glaucoma, synucleinopathies, tauopathies, frontotemporal lobar degeneration (FTLD), dementia, cadasil syndrome, hereditary cerebral hemorrhage with amyloidosis, Alexander disease, seipinopathies, familial amyloidotic neuropathy, senile systemic amyloidosis, serpinopathies, AL (light chain) amyloidosis (primary systemic amyloidosis), AH (heavy chain) amyloidosis, AA (secondary) amyloidosis, aortic medial amyloidosis, ApoAI amyloidosis, ApoAII amyloidosis, ApoAIV amyloidosis, familial amyloidosis of the Finnish type (FAF), Lysozyme amyloidosis, Fibrinogen amyloidosis, Dialysis amyloidosis, Inclusion body myositis/myopathy, Cataracts, Retinitis pigmentosa with rhodopsin mutations, medullary thyroid carcinoma, cardiac atrial amyloidosis, pituitary prolactinoma, Hereditary lattice corneal dystrophy, Cutaneous lichen amyloidosis, Mallory bodies, corneal lactoferrin amyloidosis, pulmonary alveolar proteinosis, odontogenic (Pindborg) tumor amyloid, cystic fibrosis, sickle cell disease and critical illness myopathy (CIM).
59. The method according to claim 57, wherein said peptidic compound of formula (I) is any one of SEQ ID NOs: 4-181, 196-223, 226-229, or a salt or solvate thereof.
Description
LEGENDS TO THE FIGURES
[0437]
[0438] a) Effect of SEQ ID NO: 97 and Liraglutide (all 0.02 mg/kg, s.c.) on remaining gastric contents (as indicator for gastric emptying)
[0439] b) Effect of SEQ ID NO: 97 and Liraglutide all 0.02 mg/kg, s.c., on small intestinal motility
[0440] c) Effect of SEQ ID NO: 97, at 0.02 and 0.002 mg/kg, s.c., on remaining gastric contents (as indicator for gastric emptying)
[0441] d) Effect of SEQ ID NO: 97, at 0.02 and 0.002 mg/kg, s.c., on small intestinal motility
[0442]
[0443]
[0444]
[0445]
[0446]
[0447]
[0448]
[0449]
[0450]
[0451]
[0452]
METHODS
[0453] Abbreviations employed are as follows:
[0454] ivDde: 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)3-methyl-butyl
[0455] Dde: 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)-ethyl
[0456] TFA: trifluoroacetic acid
[0457] BOP benzotriazol-1-yl-oxy-tris-(dimethylamino)-phosphonium hexafluorophosphate
[0458] HBTU 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyl-uronium hexafluorophosphate
[0459] HATU O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate
[0460] DIC N,N-diisopropylcarbodiimide
[0461] HOBt 1-hydroxybenzotriazol
[0462] DMF dimethyl formamide
[0463] EDT ethanedithiol
[0464] HPLC High Performance Liquid Chromatography
[0465] Boc tert-butyloxycarbonyl
[0466] Fmoc fluorenyloxycarbonyl
[0467] PEG Polyethylene Glycol
[0468] HTRF Homogenous Time Resolved Fluorescence
[0469] BSA bovine serum albumin
[0470] FBS fetal bovine serum
[0471] DMEM Dulbecco's modified Eagle's medium
[0472] PBS phosphate buffered saline
[0473] HEPES 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid
[0474] IBMX 3-lsobutyl-1-methylxanthine
[0475] General Synthesis of Peptidic Compounds
[0476] Materials:
[0477] Different Rink-Amide resins (4-(2,4-Dimethoxyphenyl-Fmoc-aminomethyl)-phenoxyacetamido-norleucylaminomethyl resin, Merck Biosciences; 4-[(2,4-Dimethoxyphenyl)(Fmoc-amino)methyl]phenoxy acetamido methyl resin, Agilent Technologies) were used for the synthesis of peptide amides with loadings in the range of 0.3-0.4 mmol/g. Suppliers were Merck Biosciences and Agilent Technologies. From the same suppliers 2-chloro-trityl-chloride polystyrene resins with loadings up to 1.4 mmol/g were purchased and used for the synthesis of peptide acids.
[0478] Fmoc protected natural amino acids were purchased from Protein Technologies Inc., Senn Chemicals, Merck Biosciences, Novabiochem, Iris Biotech or Bachem. The following standard amino acids were used throughout the syntheses: Fmoc-L-Ala-OH, Fmoc-L-Asn(Trt)-OH, Fmoc-L-Asp(OtBu)-OH, Fmoc-L-Cys(Trt)-OH, Fmoc-L-Gln(Trt)-OH, Fmoc-L-Glu(OtBu)-OH, Fmoc-Gly-OH, Fmoc-L-His(Trt)-OH, Fmoc-L-Ile-OH, Fmoc-L-Leu-OH, Fmoc-L-Lys(Boc)-OH, Fmoc-L-Met-OH, Fmoc-L-Phe-OH, Fmoc-L-Pro-OH, Fmoc-L-Ser(tBu)-OH, Fmoc-L-Thr(tBu)-OH, Fmoc-L-Trp(Boc)-OH, Fmoc-L-Tyr(tBu)-OH, Fmoc-L-Val-OH.
[0479] In addition, the following special amino acids were purchased from the same suppliers as above: Fmoc-L-Lys(ivDde)-OH, Fmoc-Aib-OH, Fmoc-D-Ser(tBu)-OH, Fmoc-D-Ala-OH, Boc-L-His(Boc)-OH (available as toluene solvate) and Boc-L-His(Trt)-OH.
[0480] The solid phase peptide syntheses were performed on a Prelude Peptide Synthesizer (Protein Technologies Inc) using standard Fmoc chemistry and HBTU/DIPEA activation. DMF was used as the solvent. Deprotection: 20% piperidine/DMF for 22.5 min. Washes: 7DMF. Coupling 2:5:10 200 mM AA/500 mM HBTU/2M DIPEA in DMF 2 for 20 min. Washes: 5DMF.
[0481] In cases where a Lys-side chain was modified, Fmoc-L-Lys(ivDde)-OH was used in the corresponding position. After completion of the synthesis, the ivDde group was removed according to a modified literature procedure (S. R. Chhabra et al., Tetrahedron Lett. 39, (1998), 1603), using 4% hydrazine hydrate in DMF. The following acylations were carried out by treating the resin with the N-hydroxy succinimide esters of the desired acid or using coupling reagents like HBTU/DIPEA or HOBt/DIC.
[0482] All the peptides that had been synthesized were cleaved from the resin with King's cleavage cocktail consisting of 82.5% TFA, 5% phenol, 5% water, 5% thioanisole, 2.5% EDT. The crude peptides were then precipitated in diethyl or diisopropyl ether, centrifuged, and lyophilized. Peptides were analysed by analytical HPLC and checked by ESI mass spectrometry. Crude peptides were purified by a conventional preparative HPLC purification procedure.
[0483] Analytical HPLC was performed on an Agilent 1100 Series HPLC system with a Waters XBridge BEH130 3.5 m C18 column (2.1150 mm) at 40 C. with a gradient elution at a flow rate of 0.5 mL/min and monitored at 215 and 280 nm. The gradients were set up as 10% B to 90% B over 15 min and then 90% B for 1 min or as 15% B to 50% B over 12.5 min and then 50% B to 90% B over 3 min. Buffer A=0.1% formic acid in water and B=0.1% formic acid in acetonitrile.
[0484] General Preparative HPLC Purification Procedure:
[0485] The crude peptides were purified either on an kta Purifier System or on a Jasco semiprep HPLC System. Preparative RP-C18-HPLC columns of different sizes and with different flow rates were used depending on the amount of crude peptide to be purified. Acetonitrile+0.1% TFA (B) and water+0.1% TFA (A) were employed as eluents. Product-containing fractions were collected and lyophilized to obtain the purified product.
[0486] Solubility and Stability-Testing of Exendin-4 Derivatives
[0487] Prior to the testing of solubility and stability of a peptide batch, its content was determined. Therefore, two parameters were investigated, its purity (HPLC-UV) and the amount of salt load of the batch (ion chromatography). Since synthesized peptides contain primarily trifluoroacetate anions, only anion chromatography was performed.
[0488] For solubility testing, the target concentration was 1.0 mg/mL pure compound. Therefore, solutions from solid samples were prepared in different buffer systems with a concentration of 1.0 mg/mL compound based on the previously determined content. HPLC-UV was performed after 2 h of gentle agitation from the supernatant, which was obtained by 20 min of centrifugation at 4000 rpm.
[0489] The solubility was then determined by comparison with the UV peak areas obtained with a stock solution of the peptide at a concentration of 2 mg/mL in pure water or a variable amount of acetonitrile (optical control that all of the compound was dissolved). This analysis also served as starting point (t0) for the stability testing.
[0490] For stability testing, an aliquot of the supernatant obtained for solubility was stored for 7 days at 25 C. After that time course, the sample was centrifuged for 20 min at 4000 rpm and the supernatant was analysed with HPLC-UV.
[0491] For determination of the amount of the remaining peptide, the peak areas of the target compound at t0 and t7 were compared, resulting in % remaining peptide, following the equation
% remaining peptide=[(peak area peptide t7)100]/peak area peptide t0.
[0492] The amount of soluble degradation products was calculated from the comparison of the sum of the peak areas from all observed impurities reduced by the sum of peak areas observed at t0 (i.e. to determine the amount of newly formed peptide-related species). This value was given in percentual relation to the initial amount of peptide at t0, following the equation:
% soluble degradation products={[(peak area sum of impurities t7)(peak area sum of impurities t0)]100}/peak area peptide t0
[0493] The potential difference from the sum of % remaining peptide and % soluble degradation products to 100% reflects the amount of peptide which did not remain soluble upon stress conditions following the equation
% precipitate=100-([% remaining peptide]+[% soluble degradation products])
[0494] This precipitate includes non-soluble degradation products, polymers and/or fibrils, which have been removed from analysis by centrifugation.
[0495] Anion Chromatography
[0496] Instrument: Dionex ICS-2000, pre/column: Ion Pac AG-18 250 mm (Dionex)/AS18 2250 mm (Dionex), eluent: aqueous sodium hydroxide, flow: 0.38 mL/min, gradient: 0-6 min: 22 mM KOH, 6-12 min: 22-28 mM KOH, 12-15 min: 28-50 mM KOH, 15-20 min: 22 mM, suppressor: ASRS 300 2 mm, detection: conductivity.
[0497] HPLC-UV
[0498] Instrument: Agilent 1100, column: X-Bridge C18 3.5 m 2.1150 mm (Waters), eluent: A: H20+500 ppm TFA/B: Methanol, flow: 0.55 mL/min, gradient: 0-5 min: 10-60% B; 5-15 min: 60-99% B; detection: 214 nm.
[0499] In Vitro Cellular Assays for GLP-1 Receptor and Qlucagon Receptor Efficacy
[0500] Agonism of compounds for the two receptors was determined by functional assays measuring cAMP response of HEK-293 cell lines stably expressing human GLP-1 or glucagon receptor.
[0501] cAMP content of cells was determined using a kit from Cisbio Corp. (cat. no. 62AM4PEC) based on HTRF (Homogeneous Time Resolved Fluorescence). For preparation, cells were split into T175 culture flasks and grown overnight to near confluency in medium (DMEM/10% FBS). Medium was then removed and cells washed with PBS lacking calcium and magnesium, followed by proteinase treatment with accutase (Sigma-Aldrich cat. no. A6964). Detached cells were washed and resuspended in assay buffer (1HBSS; 20 mM HEPES, 0.1% BSA, 2 mM IBMX) and cellular density determined. They were then diluted to 400000 cells/ml and 25 l-aliquots dispensed into the wells of 96-well plates. For measurement, 25 l of test compound in assay buffer was added to the wells, followed by incubation for 30 minutes at room temperature. After addition of HTRF reagents diluted in lysis buffer (kit components), the plates were incubated for 1 hr, followed by measurement of the fluorescence ratio at 665/620 nm. In vitro potency of agonists was quantified by determining the concentrations that caused 50% activation of maximal response (EC50).
[0502] Bioanalytical Screening Method for Quantification of Peptide GLP1-GCG Receptor Agonists in Mice
[0503] Mice were dosed 1 mg/kg subcutaneously (s.c.). The mice were sacrificed and blood samples were collected after 0.25, 1, 2, 4, 8, 16 and 24 hours post application. Plasma samples were analysed after protein precipitation via liquid chromatography mass spectrometry (LC/MS). PK parameters and half-life were calculated using WinonLin Version 5.2.1 (non-compartment model).
[0504] Gastric Emptying and Intestinal Passage in Mice
[0505] Female NMRI-mice of a body weight between 20 and 30 g were used. Mice were adapted to housing conditions for at least one week.
[0506] Mice were overnight fasted, while water remained available all the time. On the study day, mice were weighed, single-caged and allowed access to 500 mg of feed for 30 min, while water was removed. At the end of the 30 min feeding period, remaining feed was removed and weighed. 60 min later, a coloured, non-caloric bolus was instilled via gavage into the stomach. The test compound/reference compound or its vehicle in the control group was administered subcutaneously, to reach Cmax when coloured bolus was administered. After another 30 min, the animals were sacrificed and the stomach and the small intestine prepared. The filled stomach was weighed, emptied, carefully cleaned and dried and reweighed. The calculated stomach content indicated the degree of gastric emptying. The small intestine was straightened without force and measured in length. Then the distance from the gastric beginning of the gut to the tip of the farthest travelled intestinal content bolus was measured. The intestinal passage was given as relation in percent of the latter distance and the total length of the small intestine.
[0507] Statistical analyses were performed with Everstat 6.0 by 1-way-ANOVA, followed by Dunnetts or Newman-Keuls as post-hoc test, respectively. Differences were considered statistically significant at the p<0.05 level. As post hoc test Dunnet's Test was applied to compare versus vehicle control, only. Newman-Keul's Test was applied for all pairwise comparisons (i.e. versus vehicle and reference groups).
[0508] Automated Assessment of Feed Intake in Mice
[0509] Female NMRI-mice of a body weight between 20 and 30 g were used. Mice were adapted to housing conditions for at least one week and for at least one day single-caged in the assessment equipment, when basal data were recorded simultaneously. On the study day, test product was administered subcutaneously close to the lights-off phase (12 h lights off) and assessment of feed consumption was directly started afterwards.
[0510] Assessment included continued monitoring (every 30 min) over 22 hours. Repetition of this procedure over several days was possible. Restriction of assessment to 22 hours was for practical reasons to allow for reweighing of animals, refilling of feed and water and drug administration between procedures. Results could be assessed as cumulated data over 22 hours or differentiated to 30 min intervals.
[0511] Statistical analyses were performed with Everstat 6.0 by two-way ANOVA on repeated measures and Dunnetts post-hoc analyses. Differences were considered statistically significant at the p<0.05 level.
[0512] Acute and Subchronic Effects of Exendin-4 Derivatives after Subcutaneous Treatment on Blood Glucose and Body Weight in Female Diet-Induced Obese (DIO) C57BL/6NCrl Mice (10 Months on High-Fat Diet)
[0513] Female C57BL/6NCrl mice were housed in groups in a specific pathogen-free barrier facility on a 12-h light/dark cycle with free access to water and high-fat diet. After 10 months on high-fat diet, mice were stratified to treatment groups (n=8), so that each group had similar mean body weight.
[0514] An aged-matched group with ad-libitum access to standard chow was included as standard control group.
[0515] Before the experiment, mice were subcutaneously (s.c.) injected with vehicle solution and weighed for 3 days to acclimate them to the procedures.
[0516] 1) Acute effect on blood glucose in fed DIO mice: initial blood samples were taken just before first administration (s.c.) of vehicle (phosphate buffer solution) or the exendin-4 derivatives at doses of 3, 10, and 100 g/kg (dissolved in phosphate puffer), respectively.
[0517] The volume of administration was 5 mL/kg. The animals had access to water and their corresponding diet during the experiment, food consumption was determined at all time points of blood sampling. Blood glucose levels were measured at t=0.5 h, t=1 h, t=2 h, t=4 h, t=6 h, t=8 h, and t=24 h (method: d-glucose hexokinase, hemolysate, AU640 Beckman Coulter). Blood sampling was performed by tail incision without anaesthesia.
[0518] Comparable data can also be obtained when using male mice.
[0519] 2) Subchronic effect on body weight: all animals were treated once daily s.c. in the morning, at the beginning of the light phase (12 h lights on) with either vehicle or exendin-4 derivatives at the abovementioned doses for 4 weeks. Body weight was recorded daily. On days 6 and 28, total fat mass was measured by nuclear magnetic resonance (NMR) using a Bruker minispec (Ettlingen, Germany).
[0520] Comparable data can be obtained for both female and male mice.
[0521] Statistical analyses were performed with Everstat 6.0 by repeated measures two-way ANOVA and Dunnetts post-hoc analyses (glucose profile) and 1-way-ANOVA, followed by Dunnetts post-hoc test (body weight, body fat). Differences versus vehicle-treated DIO control mice were considered statistically significant at the p<0.05 level.
[0522] Acute and Subchronic Effects of Exendin-4 Derivatives after Subcutaneous Treatment on Blood Glucose and HbA1c in Female Leptin-Receptor Deficient Diabetic Db/Db Mice
[0523] Female BKS.Cg-m+/+Leprdb/J (db/db) and BKS.Cg-m+/+Leprdb/+(lean control) mice were obtained from Charles River Laboratories, Germany, at an age of 9-10 weeks. The animals were housed in groups in a specific pathogen-free barrier facility on a 12-h light/dark cycle with free access to water and rodent-standard chow. After 1 week of acclimatization, blood samples were drawn from the tail without anaesthesia and blood glucose (method: d-glucose hexokinase, hemolysate, AU640 Beckman Coulter) and HbA1c level (method: hemolysate, Cobas6000 c501, Roche Diagnostics, Germany) were determined.
[0524] HbA1c is a glycosylated form of haemoglobin whose level reflects the average level of glucose to which the erythrocyte has been exposed during its lifetime. In mice, HbA1c is a relevant biomarker for the average blood glucose level during the preceding 4 weeks (erythrocyte life span in mouse47 days).
[0525] Db/db mice were stratified to treatment groups (n=8), so that each group had similar baseline blood glucose and HbA1c levels.
[0526] 1) Acute effect on blood glucose in fed db/db mice: initial blood samples were taken just before first administration (s.c.) of vehicle (phosphate buffer solution) or exendin-4 derivatives at doses of 3, 10, and 100 g/kg (dissolved in phosphate puffer), respectively. The volume of administration was 5 mL/kg. The animals had access to water and chow during the experiment, food consumption was determined at all time points of blood sampling. Blood glucose levels were measured at t=0.5 h, t=1 h, t=2 h, t=4 h, t=6 h, t=8 h, and t=24 h. Blood sampling was performed by tail incision without anaesthesia.
[0527] Comparable data can also be obtained when using male mice.
[0528] 2) Subchronic effect on blood glucose and HbA1c: all animals were treated once daily s.c. with either vehicle or exendin-4 derivatives at the abovementioned doses for 4 weeks. At the end of the study, blood samples (tail, no anaesthesia) were analyzed for glucose and HbA1c.
[0529] Comparable data can be obtained for both female and male mice.
[0530] Statistical analyses were performed with Everstat 6.0 by repeated measures two-way ANOVA and Dunnetts post-hoc analyses. Differences versus vehicle-treated db/db control mice were considered statistically significant at the p<0.05 level.
EXAMPLES
[0531] The invention is further illustrated by the following examples.
Example 1
[0532] Synthesis of SEQ ID NO: 4
[0533] The solid phase synthesis was carried out on Novabiochem Rink-Amide resin (4-(2,4-Dimethoxyphenyl-Fmoc-aminomethyl)-phenoxyacetamido-norleucylaminomethyl resin), 100-200 mesh, loading of 0.34 mmol/g. The Fmoc-synthesis strategy was applied with HBTU/DIPEA-activation. In position 14 Fmoc-Lys(ivDde)-OH and in position 1 Boc-His(Boc)-OH were used in the solid phase synthesis protocol. The ivDde-group was cleaved from the peptide on resin according to a modified literature procedure (S. R. Chhabra et al., Tetrahedron Lett. 39, (1998), 1603), using 4% hydrazine hydrate in DMF. Hereafter Palm-Glu(OSu)-OtBu was coupled to the liberated amino-group. The peptide was cleaved from the resin with King's cocktail (D. S. King, C. G. Fields, G. B. Fields, Int. J. Peptide Protein Res. 36, 1990, 255-266). The crude product was purified via preparative HPLC on a Waters column (Sunfire, Prep C18) using an acetonitrile/water gradient (both buffers with 0.1% TFA).
[0534] Finally, the molecular mass of the purified peptide was confirmed by LC-MS.
Example 2
[0535] Synthesis of SEQ ID NO: 5
[0536] The solid phase synthesis was carried out on Novabiochem Rink-Amide resin (4-(2,4-Dimethoxyphenyl-Fmoc-aminomethyl)-phenoxyacetamido-norleucylaminomethyl resin), 100-200 mesh, loading of 0.34 mmol/g. The Fmoc-synthesis strategy was applied with HBTU/DIPEA-activation. In position 14 Fmoc-Lys(ivDde)-OH and in position 1 Boc-His(Boc)-OH were used in the solid phase synthesis protocol. The ivDde-group was cleaved from the peptide on resin according to a modified literature procedure (S. R. Chhabra et al., Tetrahedron Lett. 39, (1998), 1603), using 4% hydrazine hydrate in DMF. Hereafter Palm(yOSu) was coupled to the liberated amino-group. The peptide was cleaved from the resin with King's cocktail (D. S. King, C. G. Fields, G. B. Fields, Int. J. Peptide Protein Res. 36, 1990, 255-266). The crude product was purified via preparative HPLC on a Waters column (Sunfire, Prep C18) using an acetonitrile/water gradient (both buffers with 0.1% TFA).
[0537] Finally, the molecular mass of the purified peptide was confirmed by LC-MS.
Example 3
[0538] Synthesis of SEQ ID NO: 6
[0539] The solid phase synthesis was carried out on Novabiochem Rink-Amide resin (4-(2,4-Dimethoxyphenyl-Fmoc-aminomethyl)-phenoxyacetamido-norleucylaminomethyl resin), 100-200 mesh, loading of 0.34 mmol/g. The Fmoc-synthesis strategy was applied with HBTU/DIPEA-activation. In position 14 and in position 40 Fmoc-Lys(ivDde)-OH and in position 1 Boc-His(Boc)-OH were used in the solid phase synthesis protocol. The ivDde-group was cleaved from the peptide on resin according to a modified literature procedure (S. R. Chhabra et al., Tetrahedron Lett. 39, (1998), 1603), using 4% hydrazine hydrate in DMF. Hereafter Palm-Glu(OSu)-OtBu was coupled to the liberated amino-group. The peptide was cleaved from the resin with King's cocktail (D. S. King, C. G. Fields, G. B. Fields, Int. J. Peptide Protein Res. 36, 1990, 255-266). The crude product was purified via preparative HPLC on a Waters column (Sunfire, Prep C18) using an acetonitrile/water gradient (both buffers with 0.1% TFA).
[0540] Finally, the molecular mass of the purified peptide was confirmed by LC-MS.
Example 4
[0541] Synthesis of SEQ ID NO: 7
[0542] The solid phase synthesis was carried out on Novabiochem Rink-Amide resin (4-(2,4-Dimethoxyphenyl-Fmoc-aminomethyl)-phenoxyacetamido-norleucylaminomethyl resin), 100-200 mesh, loading of 0.34 mmol/g. The Fmoc-synthesis strategy was applied with HBTU/DIPEA-activation. In position 14 Fmoc-Lys(ivDde)-OH and in position 1 Boc-His(Boc)-OH were used in the solid phase synthesis protocol. The ivDde-group was cleaved from the peptide on resin according to a modified literature procedure (S. R. Chhabra et al., Tetrahedron Lett. 39, (1998), 1603), using 4% hydrazine hydrate in DMF. Hereafter Fmoc-GABA was coupled to the liberated amino-group employing the coupling reagents HBTU/DIPEA followed by Fmoc-deprotection with 20% piperidine in DMF. Finally palmitic acid was coupled to the amino-group of GABA using HBTU/DIPEA. The peptide was cleaved from the resin with King's cocktail (D. S. King, C. G. Fields, G. B. Fields, Int. J. Peptide Protein Res. 36, 1990, 255-266). The crude product was purified via preparative HPLC on a Waters column (Sunfire, Prep C18) using an acetonitrile/water gradient (both buffers with 0.1% TFA).
[0543] Finally, the molecular mass of the purified peptide was confirmed by LC-MS.
Example 5
[0544] Synthesis of SEQ ID NO: 8
[0545] The solid phase synthesis was carried out on Agilent Technologies Rink-Amide resin (4-[(2,4-Dimethoxyphenyl)(Fmoc-amino)methyl]phenoxyacetomido methyl resin), 75-150 m, loading of 0.38 mmol/g. The Fmoc-synthesis strategy was applied with HBTU/DIPEA-activation. In position 14 Fmoc-Lys(ivDde)-OH and in position 1 Boc-His(Boc)-OH were used in the solid phase synthesis protocol. The ivDde-group was cleaved from the peptide on resin according to a modified literature procedure (S. R. Chhabra et al., Tetrahedron Lett. 39, (1998), 1603), using 4% hydrazine hydrate in DMF. Hereafter Fmoc-Glu-OtBu was coupled to the liberated amino-group using HBTU/DIPEA for activation followed by the removal of the Fmoc-group with 20% piperidine in DMF. Stearic acid was coupled onto the resulting amino group after activation with HBTU/DIPEA. The peptide was cleaved from the resin with King's cocktail (D. S. King, C. G. Fields, G. B. Fields, Int. J. Peptide Protein Res. 36, 1990, 255-266). The crude product was purified via preparative HPLC on a Waters column (Sunfire, Prep C18) using an acetonitrile/water gradient (both buffers with 0.1% TFA).
[0546] Finally, the molecular mass of the purified peptide was confirmed by LC-MS.
Example 6
[0547] Synthesis of SEQ ID NO: 9
[0548] The solid phase synthesis was carried out on Agilent Technologies Rink-Amide resin (4-[(2,4-Dimethoxyphenyl)(Fmoc-amino)methyl]phenoxyacetomido methyl resin), 75-150 m, loading of 0.38 mmol/g. The Fmoc-synthesis strategy was applied with HBTU/DIPEA-activation. In position 14 Fmoc-Lys(ivDde)-OH and in position 1 Boc-His(Boc)-OH were used in the solid phase synthesis protocol. The ivDde-group was cleaved from the peptide on resin according to a modified literature procedure (S. R. Chhabra et al., Tetrahedron Lett. 39, (1998), 1603), using 4% hydrazine hydrate in DMF. Hereafter Fmoc-Glu-OtBu was coupled to the liberated amino-group using HBTU/DIPEA for activation followed by the removal of the Fmoc-group with 20% piperidine in DMF. 4-Dodecyloxy benzoic acid was coupled onto the resulting amino group after activation with HBTU/DIPEA. The peptide was cleaved from the resin with King's cocktail (D. S. King, C. G. Fields, G. B. Fields, Int. J. Peptide Protein Res. 36, 1990, 255-266). The crude product was purified via preparative HPLC on a Waters column (Sunfire, Prep C18) using an acetonitrile/water gradient (both buffers with 0.1% TFA).
[0549] Finally, the molecular mass of the purified peptide was confirmed by LC-MS.
Example 7
[0550] Synthesis of SEQ ID NO: 10
[0551] The solid phase synthesis was carried out on Agilent Technologies Cl-Trt-Cl resin (2,-Dichlorobenzhydryl-polystyrene crosslinked with divinylbenzene), 75-150 m, loading of 1.4 mmol/g. Fmoc-Ser-OAllyl was synthesized according to literature (S. Ficht, R. J. Payne, R. T. Guy, C.-H. Wong, Chem. Eur. J. 14, 2008, 3620-3629) and coupled via the side chain hydroxyl function onto Cl-Trt-Cl-resin using DIPEA in dichloromethane. The Fmoc-synthesis strategy was applied with HBTU/DIPEA-activation. In position 14 Fmoc-Lys(ivDde)-OH and in position 1 Boc-His(Boc)-OH were used in the solid phase synthesis protocol. The ivDde-group was cleaved from the peptide on resin according to a modified literature procedure (S. R. Chhabra et al., Tetrahedron Lett. 39, (1998), 1603), using 4% hydrazine hydrate in DMF. Hereafter Fmoc-Glu-OtBu was coupled to the liberated amino-group using HBTU/DIPEA for activation followed by the removal of the Fmoc-group with 20% piperidine in DMF. Palmitic acid was coupled onto the resulting amino group after activation with HBTU/DIPEA. The allyl-ester group was removed employing the procedure described in literature (S. Ficht, R. J. Payne, R. T. Guy, C.-H. Wong, Chem. Eur. J. 14, 2008, 3620-3629) followed by activation of the C-terminus with HOBt/DIC in DMF and addition of n-propylamin. The peptide was cleaved from the resin with King's cocktail (D. S. King, C. G. Fields, G. B. Fields, Int. J. Peptide Protein Res. 36, 1990, 255-266). The crude product was purified via preparative HPLC on a Waters column (Sunfire, Prep C18) using an acetonitrile/water gradient (both buffers with 0.1% TFA).
[0552] Finally, the molecular mass of the purified peptide was confirmed by LC-MS.
[0553] In an analogous way, the other peptides listed in Table 2 were synthesized.
TABLE-US-00011 TABLE 2 List of synthesized peptides and comparison of calculated vs. found molecular weight SEQ ID NO calc. mass found mass 4 4553.1 4552.4 5 4422.0 4421.4 6 5046.9 5046.8 7 4396.0 4395.1 8 4610.2 4609.8 9 4518.1 4518.2 10 4624.2 4624.6 11 4425.0 4424.4 12 4352.0 4351.2 13 4395.0 4394.1 14 4396.9 4396.0 15 4395.0 4394.4 16 4483.0 4482.0 17 4483.0 4483.2 18 4439.9 4439.1 19 4481.1 4480.5 20 4440.9 4440.0 21 4439.0 4438.2 22 4468.0 4467.9 23 4537.2 4536.5 24 4440.0 4439.5 25 4438.0 4437.4 26 4468.1 4467.2 27 4466.1 4465.3 28 4454.0 4454.0 29 4438.1 4437.3 30 4426.0 4425.9 31 4424.0 4423.9 32 4310.9 4310.3 33 4308.9 4308.3 34 4468.0 4467.9 35 4439.9 4439.4 36 4438.0 4437.3 37 4454.0 4453.9 38 4452.0 4451.9 39 4425.9 4425.9 40 4468.0 4467.4 41 4466.0 4465.4 42 4310.8 4310.3 43 4308.9 4308.3 44 4468.0 4467.4 45 4494.1 4493.4 46 4423.0 4422.3 47 4482.0 4482.0 48 4466.1 4465.4 49 4597.1 4596.4 50 4424.0 4423.5 51 4496.1 4495.2 52 4625.2 4626.0 53 4452.1 4452.0 54 4509.1 4509.0 55 4494.0 4493.7 56 4450.0 4449.6 57 4742.4 4741.6 58 4698.4 4698.0 59 4538.2 4538.3 60 4552.2 4552.1 61 4508.1 4507.7 62 4490.0 4490.2 63 4474.0 4474.3 64 4474.0 4474.3 65 4496.1 4495.5 66 4338.9 4338.4 67 4496.1 4495.7 68 4551.2 4550.5 69 4422.1 4421.5 70 4466.1 4465.5 71 4539.1 4538.8 72 4525.0 4524.8 73 4562.1 4561.5 74 4539.1 4538.4 75 4510.1 4509.4 76 4381.0 4380.3 77 4551.1 4550.5 78 4553.1 4552.7 79 4567.1 4566.7 80 4583.1 4582.4 81 4454.0 4453.5 82 4696.3 4695.8 83 4567.1 4566.7 84 4596.2 4595.4 85 4610.2 4609.7 86 4513.0 4512.8 87 4624.2 4623.4 88 4623.2 4622.5 89 4856.5 4856.3 90 4554.1 4553.7 91 4646.1 4645.8 92 4626.2 4625.5 93 4596.1 4595.4 94 4596.1 4595.3 95 4610.2 4609.5 96 4640.2 4639.8 97 4582.1 4581.7 98 4651.3 4651.1 99 4672.3 4672.1 100 4638.3 4638.0 101 4638.3 4638.2 102 4652.2 4652.2 103 4664.2 4663.7 104 4830.4 4830.3 105 5711.5 5711.2 106 4806.6 4806.5 107 4766.5 4766.0 108 4792.6 4792.6 109 4834.6 4834.5 110 4778.5 4778.9 111 4724.3 4723.9 112 4595.2 4594.7 113 4637.2 4636.7 114 4508.1 4507.7 115 4580.1 4579.4 116 4596.1 4595.4 117 4594.2 4593.4 118 4539.1 4538.6 119 4424.0 4423.4 120 4553.1 4552.5 121 4466.1 4466.0 122 4337.0 4336.5 123 4511.0 4511.0 124 4525.1 4525.0 125 4624.2 4623.7 126 4652.2 4651.7 127 4638.2 4637.7 128 4555.1 4554.3 129 4569.1 4568.6 131 4381.0 4380.9 133 4506.2 4505.4 134 4470.0 4470.0 135 4484.0 4484.0 136 4468.1 4468.0 137 4463.0 4462.4 138 4475.2 4475.8 139 4495.2 4495.6 140 4555.1 4554.0 142 4482.1 4481.4 143 4468.0 4467.0 144 4440.0 4439.1 145 4442.0 4440.0 146 4468.0 4466.1 147 4441.0 4438.8 148 4464.1 4462.2 149 4506.2 4505.4 150 4453.1 4453.6 151 4468.0 4467.9 152 4593.2 4592.1 153 4506.2 4505.1 155 4423.9 4423.9 156 4452.0 4451.9 157 4454.0 4453.9 158 4464.1 4462.8 159 4506.2 4504.8 161 4581.2 4580.7 162 4565.2 4564.2 163 4567.1 4566.4 164 4468.1 4468.0 166 4541.1 4540.8 173 4442.0 4441.9 174 4609.2 4608.3 175 4595.2 4594.8 183 4214.6 4214.1 184 4188.6 4190.7 185 4259.7 4259.0 186 4231.7 4231.0 187 4188.6 4188.4 188 4174.6 4172.0 189 4075.5 4074.8 190 4145.6 4145.1 191 4057.4 4056.2 192 4043.4 4043.4 193 4043.4 4043.2 196 4496.1 4494.4 197 4577.3 4575.6 198 4563.2 4561.2 199 4593.2 4591.2 200 4591.3 4589.7 201 4548.3 4546.2 202 4536.2 4534.0 203 4534.2 4532.4 204 4548.3 4546.2 205 4591.3 4590.4 206 4565.3 4567.0 207 4710.3 4710.6 208 4562.1 4559.6 209 4620.3 4618.8 210 4618.4 4616.1 211 4533.3 4532.4 212 4575.3 4573.5 213 4493.1 4493.4 214 4521.1 4523.4 215 4535.2 4536.9 217 4544.2 4545.0 219 4546.2 4545.3 221 4495.1 4494.4 222 4523.1 4522.4 226 4622.2 4621.6 227 4631.2 4629.6
[0554] In an analogous way, the following peptides of Table 3 can be synthesized:
TABLE-US-00012 TABLE 3 List of peptides that can be synthesized in an analogous way. SEQ ID NO 130 132 141 154 160 165 167 168 169 170 171 172 176 177 178 179 180 181 182 218 223 228 229
Example 8: Chemical Stability and Solubility
[0555] Solubility and chemical stability of peptidic compounds were assessed as described in Methods. The results are given in Table 4.
TABLE-US-00013 TABLE 4 Chemical stability and solubility Stability Solubility [mg/ml] SEQ ID NO pH 4.5 pH 7.4 pH 4.5 pH 7.4 35 100 100 >1000 >1000 36 99.7 100 >1000 >1000 44 99.1 99.4 >1000 >1000 24 100 100 >1000 >1000 25 99.6 99.6 >1000 >1000 66 100 98.1 >1000 >1000 82 98.4 99.9 >1000 >1000 126 99.5 91.4 >1000 >1000 85 95.9 85.8 >1000 >968.6 97 99.5 96.5 >2000 >2000 70 98.2 97.5 >1000 >1000 4 99.5 98.8 >815 >910 117 98.3 87.2 >1000 >1000 121 100 90.5 >1000 >980 195 0 >985
Example 9: In Vitro Data on GLP-1 and Glucagon Receptor
[0556] Potencies of peptidic compounds at the GLP-1 and glucagon receptors were determined by exposing cells expressing human glucagon receptor (hGlucagon R) or human GLP-1 receptor (hGLP-1 R) to the listed compounds at increasing concentrations and measuring the formed cAMP as described in Methods.
[0557] The results are shown in Table 5:
TABLE-US-00014 TABLE 5 EC50 values of exendin-4 derivatives at GLP-1 and Glucagon receptors (indicated in pM) SEQ ID NO EC50 hGLP-1R EC50 hGlucagon-R 2 0.7 >10000000 3 56.6 1.0 4 5 4 5 11 109 6 141 18.9 7 3.5 20.7 8 6.3 2.3 9 2.2 4.1 10 9.2 1.7 11 3.6 25.7 12 4.6 263 13 3.1 281 14 4.6 94.7 15 6.6 176 16 2.8 117 17 1.7 93.1 18 2.6 152 19 1.9 104 20 3.8 104 21 3.8 144 22 1.1 2.4 23 5.6 126 24 1.9 9.4 25 4.2 40.6 26 5.1 5.4 27 7.7 25.1 28 5.5 12.6 29 5.9 87.9 30 3.2 7 31 1.7 9.3 32 10.2 188 33 11.2 473 34 1.5 6.7 35 1.5 14.2 36 2.7 45.9 37 1.5 12.9 38 2.9 53.1 39 2.7 7.6 40 2.6 4.8 41 3.3 20.7 42 10.2 199 43 4.1 443 44 2.7 12 45 7.5 19.9 46 3.2 25.1 47 2.2 10.3 48 5.9 53.6 49 1.1 2.9 50 3.3 11.1 51 2.7 3 52 1.9 2 53 5.4 6.5 54 4.8 4 55 5.4 15.8 56 4.5 29.3 57 45 8 58 45.6 15.1 59 7.9 6.8 60 38.4 19.3 61 5.3 16 62 3.9 10.6 63 4.9 8.4 64 3.1 6.9 65 5 5.6 66 8.4 113 67 15.7 3 68 7.9 5.7 69 44.8 52.4 70 6.5 40.9 71 20.5 5.6 72 25.9 386 73 4.1 1.7 74 4.2 1.3 75 11.1 12.5 76 44.9 162 77 4.3 11.9 78 17.8 1.6 79 23.3 7.5 80 5.8 1 81 48 7.1 82 11.7 4.7 83 53.9 41.3 84 8.1 4.3 85 8.1 10.4 86 4.9 3.5 87 3 1.3 88 2.4 1.6 89 35.6 13.7 90 8.8 3.7 91 15.1 8.9 92 26 1 93 10.7 2.6 94 5.2 2.1 95 20.6 9.2 96 74.3 3.4 97 3.5 1 98 9.6 1.4 99 15.9 2.6 100 13.5 2 101 9.8 1.7 102 7.2 1.1 103 10.1 1.7 104 6.5 1.1 105 7.9 1 106 210 10.5 107 188 37.8 108 197 9 109 430 28.6 110 213 7.2 111 8.1 2.5 112 33.6 21.1 113 11.4 5.4 114 62.3 31.1 115 2.4 1.9 116 6 3.6 117 3.8 16.5 118 15.3 4.3 119 30.8 41.2 121 6.1 23.7 122 24.9 156 123 2.6 9.7 124 3 8.4 125 31.4 6.9 126 6.6 6.8 127 14.7 9.4 128 6.2 1.6 129 14.8 4.1 131 9.1 24.9 138 5.5 9.2 140 1.3 1.5 142 4.1 2.1 150 6 35.5 152 3.2 2.3 155 2.5 25.1 156 2.9 12.5 161 5 2.4 162 3.1 2.4 173 5.7 5.9 174 2.6 1.9 175 2.5 3.1 196 7.8 1.8 197 6.8 5.8 198 8.2 2.4 199 10.1 7.2 200 4.6 4.4 201 22.7 29.6 202 26.2 6.9 203 34.9 13.1 204 34.1 12.5 205 12.3 5.2 206 3.2 12.5 207 1.1 1.2 208 2.0 1.3 209 5.4 1.9 210 6.7 3.0 211 15.5 26.4 212 14.1 6.6 213 2.7 59.1 214 4.2 16.0 215 5.3 42.6 216 4.7 19.5 217 4.3 2.1 219 2.1 3.7 220 2.0 2.3 221 1.5 9.2 222 1.8 2.9 226 1.4 19.1 227 1.4 1.1
Example 10: Pharmacokinetic Testing
[0558] Pharmacokinetic profiles were determined as described in Methods. Calculated T.sub.1/2 and c.sub.max values are shown in Table 6.
TABLE-US-00015 TABLE 6 Pharmacokinetic profiles of exendin-4 derivatives. SEQ ID NO T.sub.1/2 [h] Cmax [ng/ml] 35 3.6 4910 36 3.8 5260 44 3.4 2450 24 3.7 6560 8 3.3 2680 126 1.5 3160 97 3.2 2000 4 2.8 3590 117 2.7 5000 5 1.7 3180
Example 11: Effect of SEQ ID NO: 97 on Gastric Emptying and Intestinal Passage in Female NMRI-Mice
[0559] Female NMRI-mice, weighing on average 25-30 g, received 0.02 mg/kg of SEQ ID NO: 97, Liraglutide (SEQ ID NO: 195) as reference compound, or phosphate buffered saline (vehicle control) subcutaneously, 30 min prior to the administration of the coloured bolus. 30 min later, the assessment of stomach contents and intestinal passage was done (
[0560] In another study, female NMRI-mice, weighing on average 25-30 g, were administered subcutaneously 0.02 and 0.002 mg/kg of SEQ ID NO: 97 or phosphate buffered saline (vehicle control), 30 min prior to the administration of the coloured bolus. 30 min later, the assessment of stomach contents and intestinal passage was done (
[0561] In the study with reference compound Liraglutide, SEQ ID NO: 97 reduced intestinal passage by 67% (versus 44% and 34%, respectively) and increased gastric content by 90% (versus 19% and 21%, respectively) (p<0.0001 versus vehicle control and versus comparators, 1-W-ANOVA, followed by Newman-Keul's post-hoc test) (
[0562] When SEQ ID NO: 97 was tested at 0.02 and 0.002 mg/kg, s.c. versus PBS-control, intestinal passage was reduced by 43% and 63%, respectively, and gastric content was increased by 37% and 47%, respectively (p<0.0001 versus vehicle control, 1-W-ANOVA, followed by Dunnett's post-hoc test) (
Example 12: Effect of SEQ ID NO: 97 on 22-Hours Food Intake in Female NMRI-Mice
[0563] Fed female NMRI-mice, weighing on average 25-30 g, were administered 0.01 or 0.1 mg/kg of SEQ ID NO: 97 or phosphate buffered saline (vehicle control) subcutaneously, directly prior to start of feeding monitoring (time=0 h). Lights-off phase (dark phase) started 4 hours later.
[0564] At the tested doses, SEQ ID NO: 97 demonstrated a dose-dependent reduction of feed intake, reaching 23% (p<0.0001) and 66% (p<0.0001, 2-W-ANOVA-RM, post hoc Dunnett's Test) at the end of the study, respectively (
Example 13: Acute and Subchronic Effects of SEQ ID NO: 97 after Subcutaneous Treatment on Blood Glucose and Body Weight in Female Diet-Induced Obese (DIO) C57BL/6NCrl Mice (10 Months on High Fat Diet)
[0565] 1) Glucose Profile
[0566] After blood sampling to determine the blood glucose baseline level, fed diet-induced obese female C57BL/6NCrl mice were administered 3, 10 or 100 g/kg of SEQ ID NO: 97 or phosphate buffered solution (vehicle control on standard or high-fat diet) subcutaneously. At predefined time points, more blood samples were taken to measure blood glucose and generate the blood glucose profile over 24 h.
[0567] At the tested doses, SEQ ID NO: 97 demonstrated a significant dose-dependent decrease in blood glucose compared to DIO control mice, lasting at least 8 h in the low and medium dose group and >24 h in the high dose group (p<0.0001, 2-W-ANOVA-RM, post hoc Dunnett's Test;
[0568] 2) Body Weight
[0569] Female obese C57BL/6NCrl mice were treated for 4 weeks once daily subcutaneously in the morning, at the beginning of the light phase (12 h lights on) with 3, 10 or 100 g/kg SEQ ID NO: 97 or vehicle. Body weight was recorded daily, and body fat content was determined before the start of treatment and after 4 weeks of treatment.
[0570] Treatment with SEQ ID NO: 97 reduced body weight, whereas in the high-fat diet control group an increase in body weight could be observed. These changes resulted from a decrease (or increase in the HFD control group) in body fat, as shown by the absolute changes in body fat content. These changes reached statistical significance in the medium and high dose group (*: p<0.05, 1-W-ANOVA, post hoc Dunnett's Test, Table 7).
TABLE-US-00016 TABLE 7 Weight change in DIO mice over a 4- week treatment period (mean SEM) Overall weight Body fat Example (Dose) change (g) change (g) Control standard diet 0.7 0.5 1.1 0.5 Control high-fat diet 1.3 0.5 1.0 0.4 SEQ ID NO: 97 (3 g/kg) 0.9 1.0 0.5 0.8 SEQ ID NO: 97 (10 g/kg) 3.0 1.4* 2.5 1.0* SEQ ID NO: 97 (100 g/kg) 2.3 0.9* 2.4 0.8*
Example 14: Acute and Subchronic Effects of SEQ ID NO: 97 after Subcutaneous Treatment on Blood Glucose and HbA1c in Female Leptin-Receptor Deficient Diabetic Db/Db Mice
[0571] 1. Glucose Profile
[0572] After blood sampling to determine the blood glucose baseline level, fed diabetic female db/db mice were administered 3, 10 or 100 g/kg of SEQ ID NO: 97 or phosphate buffered solution (vehicle-treated db/db control) subcutaneously. At predefined time points, more blood samples were taken to measure blood glucose and generate the blood glucose profile over 24 h.
[0573] At the tested doses, SEQ ID NO: 97 demonstrated a significant decrease in blood glucose compared to db/db control mice, lasting up to 8 h in the low and medium dose group and >24 h in the high dose group (p<0.0001 for lean control mice; p<0.01 1-8 h after treatment for low and medium dose, p<0.0002 4-24 h for high dose; 2-W-ANOVA-RM, post hoc Dunnett's Test;
[0574] 2. Blood Glucose & HbA1c
[0575] Female diabetic mice were treated for 4 weeks once daily subcutaneously with 3, 10 or 100 g/kg SEQ ID NO: 97 or vehicle. Blood glucose and HbA1c were determined before start of treatment and at the end of the study after 4 weeks of treatment.
[0576] Before treatment started, no significant differences in blood glucose levels could be detected between db/db groups, only the lean control animals had significant lower glucose levels. During the 4 weeks of treatment, glucose levels increased in the vehicle-treated db/db control group, indicating a worsening of the diabetic situation. All SEQ ID NO: 97-treated animals displayed a significant lower blood glucose level than the db control mice at the end of the study (p<0.0001 for lean control mice; p<0.01 in SEQ ID NO: 97 groups; 2-W-ANOVA-RM, post hoc Dunnett's Test;
[0577] Corresponding to blood glucose, at the beginning of the study, no significant differences in HbA1c levels could be detected between db/db groups, only the lean control animals had significant lower levels. During the 4 weeks of treatment, HbA1c increased in the vehicle-treated db/db control group, corresponding to the increasing blood glucose levels. Animals treated with high dose SEQ ID NO: 97 displayed a significant lower HbA1c level than the db control mice at the end of the study (p<0.0001, 2-W-ANOVA-RM, post hoc Dunnett's Test;
Example 15: Comparison Testing
[0578] A selection of inventive exendin-4 derivatives comprising a functionalized amino acid in position 14 has been tested versus corresponding compounds having in this position 14 a non-functionalized amino acid. The reference pair compounds and the corresponding EC50 values at GLP-1 and Glucagon receptors (indicated in pM) are given in Table 8. As shown, the inventive exendin-4 derivatives show a superior activity in comparison to the compounds with a non-functionalized amino acid in position 14.
TABLE-US-00017 TABLE 8 Comparison of exendin-4 derivatives comprising a non-functionalized amino acid in position 14 vs. exendin-4 derivatives comprising a functionalized amino acid in position 14. EC50 values at GLP-1 and Glucagon receptors are indicated in pM. EC50 EC50 residue in SEQ ID NO hGLP-1R hGlucagon-R position 14 182 5.8 419.0 M 115 2.4 1.9 K(E-x53) 183 1020.0 916.0 K 97 6.8 1.2 K(E-x53) 194 159.0 1290.0 K(Ac) 184 85.7 991.0 M 4 5.0 4.0 K(E-x53) 185 75.7 262.0 M 125 31.4 6.9 K(E-x53) 186 102.0 590.0 M 84 8.1 4.3 K(E-x53) 187 152.0 195.0 M 78 17.8 1.6 K(E-x53) 188 89.6 186.0 M 74 4.2 1.3 K(E-x53) 189 5.6 1680.0 M 24 2.0 9.8 K(E-x53) 190 21.3 1560.0 M 75 11.1 12.5 K(E-x53) 192 6.8 478 Nle 30 3.2 7.0 K(E-x53) 224 1.3 2930 L 216 4.7 19.5 K(E-x70) 225 0.7 2870 L 215 5.3 42.6 K(E-x70) (M = methionine, K = lysine, Nle = norleucine, E-x53 = (S)-4-Carboxy-4-hexadecanoylamino-butyryl-, Ac = acetate)
Example 16: Acute and Chronic Effects of SEQ ID NO: 24 after Subcutaneous Treatment on Body Weight in Male Diet-Induced Obese (DIO) C57BL/6NCrl Mice
[0579] Body Weight
[0580] Male obese C57BL/6NCrl mice were treated for 3 weeks twice daily subcutaneously with 0.5, 1.5, 5 or 15 g/kg SEQ ID NO: 24 or vehicle. Body weight was recorded daily, and body fat content was determined before the start and after 3 weeks of treatment.
[0581] Treatment with SEQ ID NO: 24 reduced body weight significantly at dosages of 1.5, 5 and 15 g/kg (*: p<0.05, 1-W-ANOVA, post hoc Dunnett's Test, Table 9,
TABLE-US-00018 TABLE 9 Weight change in DIO mice over a 3- week treatment period (mean SEM) Overall weight Body fat Example (Dose) change (g) change (g) Control standard diet 0.02 0.2 0.02 0.22 Control high-fat diet 0.5 0.3 0.8 0.3 SEQ ID NO: 24 (0.5 g/kg bid) 0.9 0.4 0.09 0.3 SEQ ID NO: 24 (1.5 g/kg bid) 6.9 0.7 3.9 0.5 SEQ ID NO: 24 (5 g/kg bid) 7.4 0.8 4.4 0.7 SEQ ID NO: 24 (15 g/kg bid) 9.1 0.7 6.7 0.4
Example 17: Acute and Chronic Effects of SEQ ID NO: 24 after Subcutaneous Treatment on Blood Glucose and HbA1c in Female Leptin-Receptor Deficient Diabetic Db/Db Mice
[0582] 1. Glucose Profile
[0583] After blood sampling to determine the blood glucose baseline level, fed diabetic female db/db mice were administered 50 g/kg of SEQ ID NO: 24 or phosphate buffered solution (vehicle-treated db/db control) twice daily subcutaneously. At predefined time points, more blood samples were taken to measure blood glucose and generate the blood glucose profile over 24 h.
[0584] At the tested dose, SEQ ID NO: 24 demonstrated a significant decrease in blood glucose compared to db/db control mice, lasting >24 h (p<0.001; 2-W-ANOVA-RM, post hoc Dunnett's Test;
[0585] 2. Blood Glucose & HbA1c
[0586] Female diabetic mice were treated for 4 weeks subcutaneously with 50 g/kg SEQ ID NO: 24 or vehicle twice daily. Blood glucose and HbA1c were determined before start of treatment and at the end of the study after 4 weeks of treatment.
[0587] Before treatment started, no significant differences in blood glucose levels could be detected between db/db groups, only the lean control animals had significant lower glucose levels. During the 4 weeks of treatment, glucose levels increased in the vehicle-treated db/db control group, indicating a worsening of the diabetic situation. The SEQ ID NO: 24-treated animals displayed a significant lower blood glucose level than the db control mice at the end of the study (p<0.01 in SEQ ID NO: 24 group; 2-W-ANOVA-RM, post hoc Dunnett's Test;
[0588] Corresponding to blood glucose, at the beginning of the study, no significant differences in HbA1c levels could be detected between db/db groups, only the lean control animals had significant lower levels. During the 4 weeks of treatment, HbA1c increased in the vehicle-treated db/db control group, corresponding to the increasing blood glucose levels. Animals treated with SEQ ID NO: 24 displayed a significantly lower HbA1c level than the db control mice at the end of the study (p<0.001, 2-W-ANOVA-RM, post hoc Dunnett's Test;
TABLE-US-00019 TABLE 10 Sequences SEQ. ID sequence 1 H-G-E-G-T-F-T-S-D-L-S-K-Q-M-E-E-E-A-V-R-L-F-I-E-W-L-K- N-G-G-P-S-S-G-A-P-P-P-S-NH2 2 H-A-E-G-T-F-T-S-D-V-S-S-Y-L-E-G-Q-A-A-K-E-I-A-W-L-V-K- G-R-NH2 3 H-S-Q-G-T-F-T-S-D-Y-S-K-Y-L-D-S-R-R-A-Q-D-V-Q-W-L-M- N-T-OH 4 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 5 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(x53)-E-S-R-R-A-Q-D-F-I-E- W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 6 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-K(E-x53)-NH2 7 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(GABA-x53)-E-S-K-A-A-Q- D-F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 8 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 9 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x75)-E-S-R-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 10 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH(n-Propyl) 11 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-E-S-Aib-A-A-Q-D- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 12 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-Aib-A-A-Aib- L-F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 13 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-Aib-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 14 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-Aib-A-A-Q-D- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 15 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-Aib-A-A-Q-L- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 16 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-E-E-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 17 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-E-E-A-A-K-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 18 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-E-E-A-A-Aib-D- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 19 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-E-E-A-A-K-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 20 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-E-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 21 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-E-A-A-Q-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 22 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-E-K-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 23 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-E-K-K-A-K-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 24 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-K-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 25 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-K-A-A-Q-D-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 26 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-E-S-K-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 27 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-E-S-K-A-A-Q-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 28 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-K-A-A-Q-E-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 29 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-K-A-A-Q-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 30 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-K-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 31 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-K-A-A-Q-D-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 32 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(x53)-E-S-K-A-A-Q-D-F-I-E- W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 33 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(x53)-E-S-K-A-A-Q-D-F-I-E- W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 34 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-E-Q-A-A-Q-D- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 35 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-Q-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 36 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-Q-A-A-Q-D-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 37 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-Q-A-A-Q-E-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 38 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-Q-A-A-Q-E-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 39 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-Q-A-A-Q-D- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 40 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-E-S-Q-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 41 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-E-S-Q-A-A-Q-D-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 42 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(x53)-E-S-Q-A-A-Q-D-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 43 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(x53)-E-S-Q-A-A-Q-D-F-I-E- W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 44 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 45 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-E-S-R-A-A-Q-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 46 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-A-A-Aib-L- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 47 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-A-A-Q-E-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 48 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-A-A-Q-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 49 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-E-x53)-E-S-R-A-A-Q- D-F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 50 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(GABA-x53)-E-S-R-A-A-Q- D-F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 51 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-E-S-R-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 52 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-E-x70)-E-S-R-A-A-Q- D-F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 53 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(GABA-x70)-E-S-R-A-A-Q- D-F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 54 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(A-A-x70)-E-S-R-A-A-Q- D-F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 55 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x74)-E-S-R-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 56 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(GABA-x74)-E-S-R-A-A-Q- D-F-1-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 57 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x60)-E-S-R-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 58 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(GABA-x60)-E-S-R-A-A-Q- D-F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 59 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x76)-E-S-R-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 60 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x77)-E-S-R-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 61 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x79)-E-S-R-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 62 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x80)-E-S-R-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 63 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x81)-E-S-R-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 64 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x82)-E-S-R-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 65 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-E-S-R-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 66 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(x53)-E-S-R-A-A-Q-D-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 67 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Aib-D- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 68 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 69 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(x53)-E-S-R-R-A-Q-L-F-I-E- W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 70 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-A-A-Q-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 71 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-Orn(E-x53)-E-S-R-R-A-Q- D-F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 72 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-Dab(E-x53)-E-S-R-R-A-Q- D-F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 73 H-dSer-H-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 74 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 75 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Aib-D- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 76 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(x53)-E-S-R-R-A-Aib-D-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 77 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 78 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-Aib-G-G-P-S-S-G-A-P-P-P-S-NH2 79 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-Aib-G-G-P-S-S-G-A-P-P-P-S-NH2 80 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-D-G-G-P-S-S-G-A-P-P-P-S-NH2 81 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(x53)-D-S-R-R-A-Q-D-F-I- E-W-L-K-D-G-G-P-S-S-G-A-P-P-P-S-NH2 82 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-E-G-G-P-S-S-G-R-P-P-P-S-NH2 83 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(x53)-E-S-R-R-A-Q-D-F-I- E-W-L-K-E-G-G-P-S-S-G-R-P-P-P-S-NH2 84 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-K-G-G-P-S-S-G-A-P-P-P-S-NH2 85 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-K-G-G-P-S-S-G-A-P-P-P-S-NH2 86 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-Q-A-A-Q-D- F-I-E-W-L-K-N-T-G-P-S-S-G-A-P-P-P-S-NH2 87 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-E-R-R-A-K-D-F- I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 88 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-K-R-R-A-Q-D-F- I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 89 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x60)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 90 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x69)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 91 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x72)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 92 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-T-G-P-S-S-G-A-P-P-P-S-NH2 93 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-A-G-P-S-S-G-A-P-P-P-S-NH2 94 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-dAla-G-P-S-S-G-A-P-P-P-S-NH2 95 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-N-A-G-P-S-S-G-A-P-P-P-S-NH2 96 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-N-T-G-P-S-S-G-A-P-P-P-S-NH2 97 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 98 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH(pyrrolidin) 99 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH(benzyl) 100 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH(tert.butyl) 101 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-N(diethyl) 102 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-N(morpholin) 103 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH(CH2CF3) 104 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH[(CH2CH2O)4- CH2CH2COOH] 105 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH[(CH2CH2O)24- CH2CH2COOH] 106 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH[(CH2)15-CH3] 107 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH[(CH2)12-OH] 108 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH[(CH2)14-CH3] 109 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH[(CH2)17-CH3] 110 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH[(CH2)13-CH3] 111 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-K-NH2 112 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(x53)-E-S-R-R-A-Q-D-F-I- E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-K-NH2 113 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-K-NH2 114 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(x53)-E-S-R-R-A-Q-D-F-I- E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-K-NH2 115 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D-F-I- E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 116 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 117 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F-I- E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 118 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Aib-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 119 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(x53)-E-S-R-R-A-Aib-D-F-I- E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 120 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Aib-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 121 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-A-A-Aib-L- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 122 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(x53)-E-S-R-A-A-Aib-L-F-I- E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 123 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-Q-A-A-Q-D- F-I-E-W-L-K-R-G-G-P-S-S-G-A-P-P-P-S-NH2 124 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-Q-A-A-Q-D- F-I-E-W-L-K-R-A-G-P-S-S-G-A-P-P-P-S-NH2 125 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-R-G-G-P-S-S-G-A-P-P-P-S-NH2 126 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-R-dAla-G-P-S-S-G-A-P-P-P-S-NH2 127 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-R-G-G-P-S-S-G-A-P-P-P-S-NH2 128 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D- F-I-E-W-L-K-S-G-G-P-S-S-G-A-P-P-P-S-NH2 129 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-S-G-G-P-S-S-G-A-P-P-P-S-NH2 130 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-E-S-Aib-A-A-Q-L- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 131 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(GABA-x70)-E-S-Aib-A-A- Q-L-F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 132 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-Aib-A-A-Q-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 133 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-D-E-K-A-A-K-L-F-I- E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 134 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(x52)-E-S-K-A-A-Q-D-F-I-E- W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 135 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(x52)-E-S-K-A-A-Q-E-F-I-E- W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 136 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(x52)-E-S-K-A-A-Q-L-F-I-E- W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 137 H-dSer-H-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-S-K-A-A-Q-D- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 138 H-dSer-H-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-S-K-A-A-Q-L-F- I-E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 139 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x76)-D-S-K-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 140 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-E-x53)-D-S-K-A-A-Q- D-F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 141 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(Phospho1)-D-S-K-A-A-Q- D-F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 142 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X95)-D-S-K-A-A-Q-D- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 143 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-S-K-A-A-Q-D- F-I-E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 144 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-K-A-Aib-Q-D- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 145 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-K-A-S-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 146 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-K-A-L-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 147 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-K-A-A-Q-D-F- I-E-W-K-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 148 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-D-S-K-A-A-Q-L-F-I- E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 149 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x76)-D-S-K-A-A-Q-L-F-I- E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 150 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-L-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 151 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-E-Q-A-A-K-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 152 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-E-Q-R-A-K-E-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 153 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-D-E-Q-A-A-K-L-F-I- E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 154 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-E-S-Q-A-A-Q-D-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 155 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-Q-A-A-Q-D-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 156 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-S-Q-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 157 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-S-Q-A-A-Q-D- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 158 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-D-S-Q-A-A-Q-L-F-I- E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 159 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x76)-D-S-Q-A-A-Q-L-F-I- E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 160 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x61)-E-S-R-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 161 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-S-R-R-A-Q-D- F-I-E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 162 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-S-R-R-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 163 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-S-R-R-A-Q-D- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 164 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-S-K-A-A-Q-D- F-I-E-W-L-K-Aib-G-G-P-S-S-G-A-P-P-P-S-NH2 165 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-E-G-G-P-S-S-G-K-P-P-P-S-NH2 166 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-S-Q-A-A-Q-D- F-I-E-W-L-K-N-T-G-P-S-S-G-A-P-P-P-S-NH2 167 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x59)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 168 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x61)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 169 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x64)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 170 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x65)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 171 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x73)-D-S-R-R-A-Q-D- F-I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 172 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-R-G-G-P-S-S-G-E-P-P-P-S-NH2 173 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-K-A-A-Q-D-F- I-E-W-L-K-S-G-G-P-S-S-G-A-P-P-P-S-NH2 174 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-E-Q-R-A-K-E-F- I-E-W-L-K-S-G-G-P-S-S-G-A-P-P-P-S-NH2 175 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-E-Q-R-A-K-D-F- I-E-W-L-K-S-G-G-P-S-S-G-A-P-P-P-S-NH2 176 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-E-Q-R-A-K-E-F- I-E-W-L-K-S-G-G-P-S-S-G-A-P-P-P-S-NH2 177 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-K-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH[(CH2CH2O)24- CH2CH2COOH] 178 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-K-A-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH[(CH2CH2P)4- CH2CH2COOH] 179 H-S-MeQ-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 180 H-S-MeQ-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 181 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-S-R-R-A-Q-D-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 182 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-M-D-S-R-R-A-Q-D-F-I-E-W-L- K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 183 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K-D-S-R-R-A-Q-D-F-I-E-W- L-K-N-G-G-P-S-S-G-A-P-P-P-S-NH2 184 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-M-E-S-R-R-A-Q-D-F-I-E-W- L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 185 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-M-D-S-R-R-A-Q-D-F-I-E-W- L-K-R-G-G-P-S-S-G-A-P-P-P-S-NH2 186 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-M-D-S-R-R-A-Q-D-F-I-E-W- L-K-K-G-G-P-S-S-G-A-P-P-P-S-NH2 187 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-M-D-S-R-R-A-Q-D-F-I-E-W- L-K-Aib-G-G-P-S-S-G-A-P-P-P-S-NH2 188 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-M-D-S-R-R-A-Q-D-F-I-E-W- L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 189 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-M-E-S-K-A-A-Q-D-F-I-E-W- L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 190 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-M-E-S-R-R-A-Aib-D-F-I-E-W- L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 191 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-Nle-E-S-Q-A-A-Q-D-F-I-E-W- L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 192 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-Nle-D-S-K-A-A-Q-D-F-I-E-W- L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 193 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-Nle-D-S-Q-A-A-Q-D-F-I-E- W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 194 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(Ac)-E-S-R-R-A-Q-D-F-I-E- W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 195 H-A-E-G-T-F-T-S-D-V-S-S-Y-L-E-G-Q-A-A-K(E-x53)-E-I-A- W-L-V-R-G-R-G-OH 196 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-D-S-K-R-A-Aib-D- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 197 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-D-E-Q-R-A-K-L-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 198 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-D-S-R-R-A-Q-L-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 199 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-D-E-Q-R-A-K-D-F-I- E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 200 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-D-E-Q-R-A-K-L-F-I- E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 201 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x76)-D-E-Q-A-A-K-L-F-I- E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 202 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x76)-E-S-R-A-A-Q-D-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 203 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x76)-E-S-R-A-A-Q-L-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 204 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x76)-E-S-R-A-A-Q-L-F-I- E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 205 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-E-S-R-R-A-Q-L-F-I- E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 206 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-D-E-Q-K-A-K-L-F-I- E-W-L-K-S-G-G-P-S-S-G-A-P-P-P-S-NH2 207 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-E-x53)-D-E-Q-R-A-K-E- F-I-E-W-L-K-S-G-G-P-S-S-G-A-P-P-P-S-NH2 208 H-S-H-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-E-S-R-R-A-Q-D-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 209 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-D-K-R-R-A-Q-D-F-I- E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 210 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-D-K-R-R-A-Q-L-F-I- E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 211 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-D-K-R-A-A-Q-L-F-I- E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 212 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x76)-D-K-R-A-A-Q-L-F-I- E-W-L-K-A-dAla-G-P-S-S-G-A-P-P-P-S-NH2 213 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-D-E-E-A-A-K-L-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 214 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-E-E-A-A-R-L-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 215 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-E-E-E-A-A-R-L-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 216 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-E-E-A-A-R-L-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 217 H-Aib-H-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-E-E-E-A-A-R-L-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 218 H-Aib-H-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-E-E-A-A-R-L-F-I- E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 219 H-dSer-H-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-E-E-E-A-A-R-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 220 H-dSer-H-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-E-E-A-A-R-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 221 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x53)-D-E-E-A-A-R-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 222 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-X70)-D-E-E-A-A-R-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 223 H-dSer-Q-G-T-F-T-S-D-L-S-K-Q-K(E-x70)-E-E-E-A-A-R-L-F- I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 224 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-L-D-E-E-A-A-R-L-F-I-E-W-L-K- A-G-G-P-S-S-G-A-P-P-P-S-NH2 225 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-L-E-E-E-A-A-R-L-F-I-E-W-L-K- A-G-G-P-S-S-G-A-P-P-P-S-NH2 226 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-E-x53)-D-E-E-A-A-R-L- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 227 H-Aib-H-G-T-F-T-S-D-L-S-K-Q-K(E-E-x53)-D-E-E-A-A-R-L- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 228 H-Aib-Q-G-T-F-T-S-D-L-S-K-Q-K(E-E-x53)-E-E-E-A-A-R-L- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2 229 H-Aib-H-G-T-F-T-S-D-L-S-K-Q-K(E-E-x53)-E-E-E-A-A-R-L- F-I-E-W-L-K-A-G-G-P-S-S-G-A-P-P-P-S-NH2