GIP peptide analogues
Provided herein are glucose-dependent insulinotropic polypeptide (GIP)-derived peptide analogues, for example GIP(3-30), and their use as antagonists of the GIP receptor and for treatment of disorders such as obesity, diabetes mellitus and insulin resistance.
1. A method of inhibiting or reducing one or more of: i) gastric inhibitory polypeptide (GIP)-induced glucagon secretion, ii) GIP-induced insulin secretion, iii) GIP-induced somatostatin secretion, iv) GIP-induced glucose uptake, v) GIP-induced fatty acid synthesis and/or fatty acid incorporation, vi) high or increased expression or activity of a GIPR, vii) post-prandial GIP release, viii) serum levels of free fatty acids and/or triglycerides, and ix) GIP-induced reduction of bone resorption,
said method comprising one or more steps of administering to an individual in need thereof a peptide selected from the group consisting of:
a peptide consisting of 28 contiguous amino acids of amino acid sequence EGTFISDYSIAMX oa X ob IX 1 QQDFVNWLLAQX 2 (GIP3-30; SEQ ID NO: 74):
a peptide consisting of 26 contiguous amino acids of amino acid sequence TFISDYSIAMX oa X ob IX 1 QQDFVNWLLAQX 2 (GIP5-30; SEQ ID NO: 76), wherein X oa is selected from the group consisting of D, E,
X ob is selected from the group consisting of K, A, H, R,
X 1 is selected from the group consisting of H, R, A, K, F, Y and
X 2 is selected from the group consisting of K, R, H, A; and
a variant of any one of the above peptides, wherein the amino acid sequence of the variant differs from SEQ ID NO: 74 or SEQ ID NO: 76, only in that the amino acid sequence of the variant comprises one or two amino acid substitutions, wherein said substitutions are selected from the group consisting of: i) substitution of an amino acid having a polar side chain for a different amino acid having a polar side chain wherein amino acids having a polar side chain are Asp, Glu, Lys, Arg, His, Asn, Gln, Ser, Thr, Tyr, and Cys; ii) substitution of an amino acid having a non-polar side chain for a different amino acid having a non-polar side chain wherein amino acids having a non-polar side chain are Gly, Ala, Val, Leu, Ile, Phe, Trp, Pro, and Met; iii) substitution of an amino acid having an aliphatic side chain for a different amino acid having an aliphatic side chain wherein amino acids having a aliphatic side chain are Gly, Ala Val, Leu, and Ile; iv) substitution of an amino acid having a cyclic side chain for a different amino acid having a cyclic side chain wherein amino acids having a cyclic side chain are Phe, Tyr, Trp, His, and Pro; v) substitution of an amino acid having an aromatic side chain for a different amino acid having an aromatic side chain wherein amino acids having an aromatic side chain are Phe, Tyr, and Trp; vi) substitution of an amino acid having an acidic side chain for a different amino acid having an acidic side chain wherein amino acids having an acidic side chain are Asp, and Glu; vii) substitution of an amino acid having a basic side chain for a different amino acid having a basic side chain wherein amino acids having a basic side chain are Lys, Arg, and His; viii) substitution of an amino acid having an amide side chain for a different amino acid having an amide side chain wherein amino acids having an amide side chain are Asn, and Gln; ix) substitution of an amino acid having a hydroxy side chain for a different amino acid having a hydroxy side chain wherein amino acids having a hydroxy side chain are Ser, and Thr; x) substitution of an amino acid having a sulphur-containing side chain for a different amino acid having a sulphur-containing side chain wherein amino acids having a sulphur-containing side chain are Cys, and Met; xi) substitution of a neutral, weakly hydrophobic amino acid for a different neutral, weakly hydrophobic amino acid wherein neutral, weakly hydrophobic amino acids are Pro, Ala, Gly, Ser, and Thr; xii) substitution of a hydrophilic, acidic amino acid for a different hydrophilic, acidic amino acid wherein hydrophilic, acidic amino acids are Gln, Asn, Glu, and Asp, and xiii) substitution of a hydrophobic amino acid for a different hydrophobic amino acid wherein hydrophobic amino acids are Leu, Ile, and Val; wherein said peptide or variant thereof is an antagonist of a GIP receptor (GIPR).
2. The method according to claim 1 , wherein said peptide is selected from the group consisting of:
a peptide consisting of 28 contiguous amino acids of sequence EGTFISDYSIAMDKIX 1 QQDFVNWLLAQX 2 (GIP3-30; SEQ ID NO: 11),
a peptide consisting of 26 contiguous amino acids of sequence TFISDYSIAMDKIX 1 QQDFVNWLLAQX 2 (GIP5-30; SEQ ID NO: 45),
wherein X 1 is selected from the group consisting of H, R, A, K, F, Y, and
X 2 is selected from the group consisting of K, R, H, A, and
a variant of any one of the above peptides, wherein the amino acid sequence of the variant differs from SEQ ID NO: 11 or SEQ ID NO: 45, only in that the amino acid sequence of the variant comprises the one or two amino acid substitutions,
wherein said peptide or variant thereof is an antagonist of a GIP receptor (GIPR).
3. The method according to claim 1 , wherein said peptide is selected from the group consisting of:
a peptide consisting of 28 contiguous amino acids of sequence EGTFISDYSIAMDKIHQQDFVNWLLAQK (hGIP3-30, SEQ ID NO: 1);
a peptide consisting of 28 contiguous amino acids of sequence EGTFISDYSIAMDKIRQQDFVNWLLAQK (rGIP3-30, SEQ ID NO: 2);
a peptide consisting of 28 contiguous amino acids of sequence: EGTFISDYSIAMDKIRQQDFVNWLLAQR (mGIP3-30, SEQ ID NO: 3);
a peptide a peptide consisting of 26 contiguous amino acids of sequence TFISDYSIAMDKIHQQDFVNWLLAQK (hGIP5-30, SEQ ID NO: 78); and
a variant of any one of the above peptides, wherein the amino acid sequence of the variant differs from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 78, only in that the amino acid sequence of the variant comprises the one or two amino acid substitutions,
wherein said peptide or variant thereof is an antagonist of a GIP receptor (GIPR).
4. The method according to claim 1 , wherein said peptide is selected from the group consisting of:
(hGIP3-30, SEQ ID NO: 1)
EGTFISDYSIAMDKIHQQDFVNWLLAQK,
(rGIP3-30, SEQ ID NO: 2)
EGTFISDYSIAMDKIRQQDFVNWLLAQK,
(mGIP3-30, SEQ ID NO: 3)
EGTFISDYSIAMDKIRQQDFVNWLLAQR,
(hGIP5-30, SEQ ID NO: 78)
TFISDYSIAMDKIHQQDFVNWLLAQK,
(hGIP(3-30)H18A; SEQ ID NO: 79)
EGTFISDYSIAMDKI A QQDFVNWLLAQK,
(hGIP(3-30)H18K; SEQ ID NO: 80)
EGTFISDYSIAMDKI K QQDFVNWLLAQ,
(hGIP(3-30)D15EH18A; SEQ ID NO: 81)
EGTFISDYSIAM E KI A QQDFVNWLLAQK,
(hGIP(3-30)K16AH18A; SEQ ID NO: 82)
EGTFISDYSIAMD A I A QQDFVNWLLAQK,
(hGIP(3-30)D15E; SEQ ID NO: 83)
EGTFISDYSIAM E KIHQQDFVNWLLAQK,
(hGIP(3-30)D15N; SEQ ID NO: 84)
EGTFISDYSIAM N KIHQQDFVNWLLAQK,
(hGIP(3-30)K16A; SEQ ID NO: 85)
EGTFISDYSIAMD A IHQQDFVNWLLAQK,
(hGIP(3-30)K16H; SEQ ID NO: 86)
EGTFISDYSIAMD H IHQQDFVNWLLAQK,
(hGIP(3-30)K16R; SEQ ID NO: 87)
EGTFISDYSIAMD R IHQQDFVNWLLAQK,
(hGIP(3-30)H18F; SEQ ID NO: 88)
EGTFISDYSIAMDKI F QQDFVNWLLAQK,
(hGIP(3-30)H18W; SEQ ID NO: 89)
EGTFISDYSIAMDKI W QQDFVNWLLAQK,
(hGIP(3-30)K30R SEQ ID NO: 90)
EGTFISDYSIAMDKIHQQDFVNWLLAQ R ,
and
(hGIP(3-30)K30H; SEQ ID NO: 91)
EGTFISDYSIAMDKIHQQDFVNWLLAQ H .
5. The method according to claim 1 , wherein said peptide is C-terminally amidated (—NH 2 ) and/or N-terminally acetylated (COCH 3 ).
6. The method of claim 5 , wherein said peptide is selected from the group consisting of: EGTFISDYSIAMDKIHQQDFVNWLLAQK-NH 2 (SEQ ID NO:92); TFISDYSIAMDKIHQQDFVNWLLAQK-NH 2 (SEQ ID NO:94);
and a variant of any one of the above peptides, wherein the amino acid sequence of the variant differs from SEQ ID NO: 92 or SEQ ID NO: 94, only in that the amino acid sequence of the variant comprises the one or two amino acid substitutions, wherein said peptide or variant thereof is an antagonist of a GIP receptor (GIPR).
7. The method of claim 5 , wherein said peptide is selected from the group consisting of:
EGTFISDYSIAMDKIHQQDFVNWLLAQK-NH 2 (SEQ ID NO:92); and
TFISDYSIAMDKIHQQDFVNWLLAQK-NH 2 (SEQ ID NO:94).
8. The method according to claim 1 , wherein said peptide
i) binds to a GIP-receptor (GIPR),
ii) antagonizes GIP1-42-induced activation of a GIPR,
iii) displaces GIP1-42 from a GIPR,
iv) is a competitive antagonist of a GIP-receptor,
v) has an affinity for a GIPR which is higher than the affinity of GIP3-42 for the same GIPR,
vi) is capable of inhibiting and/or antagonizing somatostatin secretion induced by native GIP,
vii) is capable of inhibiting and/or antagonizing insulin secretion induced by native GIP, and/or
viii) is capable of inhibiting and/or antagonizing glucagon secretion induced by native GIP.
9. The method according to claim 1 , wherein said peptide is encoded by a nucleic acid construct.
10. A method of treating one or more of: metabolic syndrome, obesity, pre-diabetes, diabetes mellitus type 2, insulin resistance, elevated fasting glucose, elevated fasting serum triglyceride level, low high-density lipoprotein (HDL) levels, fatty acid metabolism disorder, cardiovascular disease, elevated blood pressure and atherosclerosis,
said method comprising one or more steps of administering to an individual in need thereof a peptide selected from the group consisting of:
a peptide consisting of 28 contiguous amino acids of amino acid sequence EGTFISDYSIAMX oa X ob IX 1 QQDFVNWLLAQX 2 (GIP3-30; SEQ ID NO: 74);
a peptide consisting of 26 contiguous amino acids of amino acid sequence TFISDYSIAMX oa X ob IX 1 QQDFVNWLLAQX 2 (GIP5-30; SEQ ID NO: 76), wherein X oa is selected from the group consisting of D, E,
X ob is selected from the group consisting of K, A, H, R,
X 1 is selected from the group consisting of H, R, A, K, F, Y and
X 2 is selected from the group consisting of K, R, H, A; and
a variant of any one of the above peptides, wherein the amino acid sequence of the variant differs from SEQ ID NO: 74 or SEQ ID NO: 76, only in that the amino acid sequence of the variant comprises one or two amino acid substitutions, wherein said substitutions are selected from the group consisting of: i) substitution of an amino acid having a polar side chain for a different amino acid having a polar side chain wherein amino acids having a polar side chain are Asp, Glu, Lys, Arg, His, Asn, Gln, Ser, Thr, Tyr, and Cys; ii) substitution of an amino acid having a non-polar side chain for a different amino acid having a non-polar side chain wherein amino acids having a non-polar side chain are Gly, Ala, Val, Leu, Ile, Phe, Trp, Pro, and Met; iii) substitution of an amino acid having an aliphatic side chain for a different amino acid having an aliphatic side chain wherein amino acids having a aliphatic side chain are Gly, Ala Val, Leu, and Ile; iv) substitution of an amino acid having a cyclic side chain for a different amino acid having a cyclic side chain wherein amino acids having a cyclic side chain are Phe, Tyr, Trp, His, and Pro; v) substitution of an amino acid having an aromatic side chain for a different amino acid having an aromatic side chain wherein amino acids having an aromatic side chain are Phe, Tyr, and Trp; vi) substitution of an amino acid having an acidic side chain for a different amino acid having an acidic side chain wherein amino acids having an acidic side chain are Asp, and Gu; vii) substitution of an amino acid having a basic side chain for a different amino acid having a basic side chain wherein amino acids having a basic side chain are Lys, Arg, and His; viii) substitution of an amino acid having an amide side chain for a different amino acid having an amide side chain wherein amino acids having an amide side chain are Asn, and Gln; ix) substitution of an amino acid having a hydroxy side chain for a different amino acid having a hydroxy side chain wherein amino acids having a hydroxy side chain are Ser, and Thr; x) substitution of an amino acid having a sulphur-containing side chain for a different amino acid having a sulphur-containing side chain wherein amino acids having a sulphur-containing side chain are Cys, and Met; xi) substitution of a neutral, weakly hydrophobic amino acid for a different neutral, weakly hydrophobic amino acid wherein neutral, weakly hydrophobic amino acids are Pro, Ala, Gly, Ser, and Thr; xii) substitution of a hydrophilic, acidic amino acid for a different hydrophilic, acidic amino acid wherein hydrophilic, acidic amino acids are Gln, Asn, Glu, and Asp, and xii) substitution of a hydrophobic amino acid for a different hydrophobic amino acid wherein hydrophobic amino acids are Leu, Ile, and Val; wherein said peptide or variant thereof is an antagonist of a GIP receptor (GIPR).
11. The method according to claim 10 , wherein said peptide is selected from the group consisting of:
a peptide consisting of 28 contiguous amino acids of sequence EGTFISDYSIAMDKIX 1 QQDFVNWLLAQX 2 (GIP3-30; SEQ ID NO: 11),
a peptide consisting of 26 contiguous amino acids of sequence TFISDYSIAMDKIX 1 QQDFVNWLLAQX 2 (GIP5-30; SEQ ID NO: 45),
wherein X 1 is selected from the group consisting of H, R, A, K, F, Y, and
X 2 is selected from the group consisting of K, R, H, A, and
a variant of any one of the above peptides, wherein the amino acid sequence of the variant differs from SEQ ID NO: 11 or SEQ ID NO: 45, only in that the amino acid sequence of the variant comprises the one or two amino acid substitutions,
wherein said peptide or variant thereof is an antagonist of a GIP receptor (GIPR).
12. The method according to claim 10 , wherein said peptide is selected from the group consisting of:
a peptide consisting of 28 contiguous amino acids of sequence EGTFISDYSIAMDKIHQQDFVNWLLAQK (hGIP3-30, SEQ ID NO: 1);
a peptide consisting of 28 contiguous amino acids of sequence EGTFISDYSIAMDKIRQQDFVNWLLAQK (rGIP3-30, SEQ ID NO: 2);
a peptide consisting of 28 contiguous amino acids of sequence EGTFISDYSIAMDKIRQQDFVNWLLAQR (mGIP3-30, SEQ ID NO: 3);
a peptide consisting of 26 contiguous amino acids of sequence TFISDYSIAMDKIHQQDFVNWLLAQK (hGIP5-30, SEQ ID NO: 78); and
a variant of any one of the above peptides, wherein the amino acid sequence of the variant differs from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO:3 or SEQ ID NO: 78, only in that the amino acid sequence of the variant comprises the one or two amino acid substitutions,
wherein said peptide or variant thereof is an antagonist of a GIP receptor (GIPR).
13. The method according to claim 10 , wherein said peptide is selected from the group consisting of:
(hGIP3-30, SEQ ID NO: 1)
EGTFISDYSIAMDKIHQQDFVNWLLAQK,
(rGIP3-30, SEQ ID NO: 2)
EGTFISDYSIAMDKIRQQDFVNWLLAQK,
(mGIP3-30, SEQ ID NO: 3)
EGTFISDYSIAMDKIRQQDFVNWLLAQR,
(hGIP5-30, SEQ ID NO: 78)
TFISDYSIAMDKIHQQDFVNWLLAQK,
(hGIP(3-30)H18A; SEQ ID NO: 79)
EGTFISDYSIAMDKI A QQDFVNWLLAQK,
(hGIP(3-30)H18K; SEQ ID NO: 80)
EGTFISDYSIAMDKI K QQDFVNWLLAQ,
(hGIP(3-30)D15EH18A; SEQ ID NO: 81)
EGTFISDYSIAM E KI A QQDFVNWLLAQK,
(hGIP(3-30)K16AH18A; SEQ ID NO: 82)
EGTFISDYSIAMD A I A QQDFVNWLLAQK,
(hGIP(3-30)D15E; SEQ ID NO: 83)
EGTFISDYSIAM E KIHQQDFVNWLLAQK,
(hGIP(3-30)D15N; SEQ ID NO: 84)
EGTFISDYSIAM N KIHQQDFVNWLLAQK,
(hGIP(3-30)K16A; SEQ ID NO: 85)
EGTFISDYSIAMD A IHQQDFVNWLLAQK,
(hGIP(3-30)K16H; SEQ ID NO: 86)
EGTFISDYSIAMD H IHQQDFVNWLLAQK,
(hGIP(3-30)K16R; SEQ ID NO: 87)
EGTFISDYSIAMD R IHQQDFVNWLLAQK,
(hGIP(3-30)H18F; SEQ ID NO: 88)
EGTFISDYSIAMDKI F QQDFVNWLLAQK,
(hGIP(3-30)H18W; SEQ ID NO: 89)
EGTFISDYSIAMDKI W QQDFVNWLLAQK,
(hGIP(3-30)K30R SEQ ID NO: 90)
EGTFISDYSIAMDKIHQQDFVNWLLAQ R ,
and
(hGIP(3-30)K30H; SEQ ID NO: 91)
EGTFISDYSIAMDKIHQQDFVNWLLAQ H .
14. The method according to claim 10 , wherein said peptide is C-terminally amidated (—NH 2 ) and/or N-terminally acetylated (COCH 3 ).
15. The method according to claim 10 , wherein said peptide
i) binds to a GIP-receptor (GIPR),
ii) antagonises GIP1-42-induced activation of a GIPR,
iii) displaces GIP1-42 from a GIPR,
iv) is a competitive antagonist of a GIP-receptor,
v) has an affinity for a GIPR which is higher than the affinity of GIP3-42 for the same GIPR,
vi) is capable of inhibiting and/or antagonising somatostatin secretion induced by native GIP,
vii) is capable of inhibiting and/or antagonising insulin secretion induced by native GIP, and/or
viii) is capable of inhibiting and/or antagonising glucagon secretion induced by native GIP.
16. The method according to claim 10 , wherein said peptide is encoded by a nucleic acid construct.
17. The method of claim 10 , wherein said peptide is selected from the group consisting of: EGTFISDYSIAMDKIHQQDFVNWLLAQK-NH 2 (SEQ ID NO:92): TFISDYSIAMDKIHQQDFVNWLLAQK-NH 2 (SEQ ID NO:94); and a variant of any one of the above peptides, wherein the amino acid sequence of the variant differs from SEQ ID NO: 92 or SEQ ID NO: 94, only in that the amino acid sequence of the variant comprises the one or two amino acid substitutions, wherein said peptide or variant thereof is an antagonist of a GIP receptor (GIPR).
18. The method of claim 10 , wherein said peptide is selected from the group consisting of:
EGTFISDYSIAMDKIHQQDFVNWLLAQK-NH 2 (SEQ ID NO:92); and
TFISDYSIAMDKIHQQDFVNWLLAQK-NH 2 (SEQ ID NO:94).