Glucagon analogs exhibiting gip receptor activity

An analog of glucagon (SEQ ID NO:

• 1), comprising:
• a. an amino acid comprising an imidazole side chain at position 1;
• b. an DPP-IV protective amino acid at position 2, optionally, AIB;
• c. an acylated amino acid or alkylated amino acid at any of positions 9, 10, 12, 16, 20, or 37-43, optionally wherein the acyl or alkyl group is linked to the amino acid via a spacer;
• d. an alpha helix stabilizing amino acid at one or more of positions 16-21, optionally, position 16, wherein the analog does not comprise an alpha helix stabilizing amino acid at position 20;
• and
• e. up to ten additional amino acid modifications relative to SEQ ID NO:
• 1;
• wherein, when the glucagon analog lacks a hydrophilic moiety, the glucagon analog exhibits a GIP percentage potency of at least 0.1%, wherein the glucagon analog has less than 100-fold selectivity for the human GLP-1 receptor versus the GIP receptor.

A peptide comprising

• (a) the sequence of,
• (SEQ ID NO:
• 184)
• HX2X3GTFTSDX10SKYLDX16RX18AX20X21FVQWLX27X28X29GP
• SSGX35PPPS
• wherein:
• X2 is AIB;
• X3 is Gln or Gln analog;
• X10 is Tyr or an amino acid covalently attached to a C12 to C18 acyl or alkyl group;
• X16 is any amino acid, optionally, any amino acid other than Gly, Pro, and Ser;
• X18 is Arg or Ala;
• X20 is negative charged amino acid or a charge-neutral amino acid, optionally, AIB or Gln;
• X21 is an acidic amino acid, optionally, Asp or Glu;
• X27 is Leu, Ala, Nle, or Met;
• X28 is Ala or an acidic amino acid (optionally, Asp or Glu);
• X29 is Ala or Gly;
• X35 is Ala or a basic amino acid (optionally, Arg or Lys);
• wherein, when X28 is an acidic amino acid, X35 is a basic amino acid;
• wherein, when X10 is Tyr, the peptide comprises at position 40 an amino acid covalently attached to a C12 to C18 acyl or alkyl group, and, wherein, optionally, the peptide comprises Gly at position 41,
• wherein the C-terminal amino acid of the peptide is amidated;
• or
• (b) SEQ ID NO:
• 184 with up to 3 amino acid modifications relative to SEQ ID NO:
• 184, wherein the peptide exhibits agonist activity at each of the human GIP receptor, the human GLP-1 receptor and the human glucagon receptor.
• or
• (c) the sequence of:
• (SEQ ID NO:
• 185)
• HX2QGTFTSDX10SKYLDX16RX18AX20X21FVQWLX27X28X29GP
• SSGAPPPS
• wherein:
• X2 is AIB;
• X10 is Tyr or an amino acid covalently attached to a C12 to C18 acyl or alkyl group;
• X16 is Glu, an alpha, alpha disubstituted amino acid, Lys or
• X18 is Arg or Ala;
• X20 is AIB or Gln;
• X21 is Asp or Glu;
• X27 is Leu, Nle, or Met;
• X28 is Ala, Asp or Glu;
• X29 is Gly of Thr;
• and
• wherein, when X10 is Tyr, the peptide comprises at position 40 an amino acid covalently attached to a C12 to C18 acyl or alkyl group, and, wherein, optionally, the peptide comprises Gly at position 41,
• wherein the C-terminal amino acid of the peptide is amidated;
• or
• (d) SEQ ID NO:
• 185 with up to 3 amino acid modifications relative to SEQ ID NO:
• 185, wherein the peptide exhibits agonist activity at each of the human GIP receptor, the human GLP-1 receptor and the human glucagon receptor.
• or
• (e) the sequence of:
• (SEQ ID NO:
• 186)
• HX2X3GTFTSDX10SKYLDX16RX18AX20X21FVQWLX27X28X29
• wherein:
• X2 is AIB;
• X3 is Gln or Gln analog;
• X10 is Tyr or an amino acid covalently attached to a C10 to C26 acyl or alkyl group;
• X16 is any amino acid optionally, any amino acid other than Gly, Pro, and Ser;
• X18 is Arg or Ala;
• X20 is a negative charged amino acid or a charge-neutral amino acid, optionally, AIB or Gln;
• X21 is X71 is an acidic amino acid, optionally, Asp or Glu;
• X27 is Leu, Ala, Nle, or Met;
• X28 is Ala or an acidic amino acid (optionally, Asp or Glu);
• X29 is Ala, Gly or Thr;
• and
• wherein the peptide comprises an amino acid covalently attached to a C10 to C26 acyl or alkyl group, optionally, at position 10, and the C-terminal amino acid of the peptide is amidated;
• or
• (f) SEQ ID NO:
• 186 with up to 3 amino acid modifications relative to SEQ ID NO:
• 186, wherein the peptide exhibits agonist activity at each of the human GIP receptor, the human GLP-1 receptor and the human glucagon receptor
• or
• (g) the sequence of:
• (SEQ ID NO:
• 187)
• HX2QGTFTSDX10SKYLDX16RX18AX20X21FVQWLX27X28X29
• wherein:
• X2 is AIB;
• X10 is Tyr or an amino acid covalently attached to a C10 to C26 acyl or alkyl group;
• X16 is Glu, alpha, alpha-disubstituted amino acid, or Lys.
• X18 is Arg or Ala;
• X20 is a negative charged amino acid or a charge-neutral amino acid, optionally, AIB or Gln;
• X21 is Asp or Glu;
• X27 is Leu, Ala, Nle, or Met;
• X28 is Ala, Asp or Glu;
• X29 is Gly or Thr;
• and
• wherein the peptide comprises an amino acid covalently attached to a C12 to C18 acyl or alkyl group, optionally, at position 10, and the C-terminal amino acid of the peptide is amidated;
• or
• (h) SEQ ID NO:
• 187 with up to 3 amino acid modifications relative to SEQ ID NO:
• 187, wherein the peptide exhibits agonist activity at each of the human GIP receptor, the human GLP-1 receptor and the human glucagon receptor.

A peptide comprising

• (i) the sequence of SEQ ID NO:
• 188, or a sequence of SEQ ID NO:
• 188 with up to 3 amino acid modifications relative to SEQ ID NO:
• 188, wherein the peptide exhibits agonist activity at each of the human GIP receptor and the human GLP-1 receptor;
• (ii) the sequence of SEQ ID NO:
• 189, or a sequence of SEQ ID NO:
• 189 with up to 3 amino acid modifications relative to SEQ ID NO:
• 189, wherein the peptide exhibits agonist activity at each of the human GIP receptor and the human GLP-1 receptor;
• (iii) the sequence of SEQ ID NO:
• 190, or a sequence of SEQ ID NO:
• 190 with up to 3 amino acid modifications relative to SEQ ID NO:
• 190, wherein the peptide exhibits agonist activity at each of the human GIP receptor and the human GLP-1 receptor;
• (iv) the sequence of SEQ ID NO:
• 191, or a sequence of SEQ ID NO:
• 191 with up to 3 amino acid modifications relative to SEQ ID NO:
• 191, wherein the peptide exhibits agonist activity at each of the human GIP receptor and the human GLP-1 receptor;
• (v) the sequence of SEQ ID NO:
• 192, or a sequence of SEQ ID NO:
• 192 with up to 3 amino acid modifications relative to SEQ ID NO:
• 192, wherein the peptide exhibits agonist activity at each of the human GIP receptor and the human GLP-1 receptor, and optionally, wherein the peptide exhibits agonist activity at the human glucagon receptor;
• (vi) the sequence of SEQ ID NO:
• 193, or a sequence of SEQ ID NO:
• 193 with up to 3 amino acid modifications relative to SEQ ID NO:
• 193, wherein the peptide exhibits agonist activity at each of the human GIP receptor and the human GLP-1 receptor, optionally, wherein the peptide exhibits agonist activity at the human glucagon receptor.
• (vii) the sequence of SEQ ID NO:
• 194, or a sequence of SEQ ID NO:
• 194 with up to 3 amino acid modifications relative to SEQ ID NO:
• 194, wherein the peptide exhibits agonist activity at each of the human GIP receptor and the human GLP-1 receptor, optionally, wherein the peptide exhibits agonist activity at the human glucagon receptor;
• or
• (viii) the sequence of SEQ ID NO:
• 195, or a sequence of SEQ ID NO:
• 195 with up to 3 amino acid modifications relative to SEQ ID NO:
• 195, wherein the peptide exhibits agonist activity at each of the human GIP receptor and the human GLP-1 receptor, optionally, wherein the peptide exhibits agonist activity at the human glucagon receptor.

A peptide comprising the sequence of SEQ ID NO:

• 37;
• SEQ ID NO:
• 171 or SEQ ID NO:
• 180.

An analog of glucagon (SEQ ID NO:

• 1) having GIP agonist activity, comprising:
• (a) an amino acid comprising an imidazole side chain at position 1,
• (b) at position 16, an amino acid of Formula IV:
• wherein n is 1 to 7, wherein each of R1 and R2 is independently selected from the group consisting of H, C1-C18 alkyl, (C1-C18 alkyl)OH, (C1-C18 alkyl)NH2, (C1-C18 alkyl)SH, (C0-C4 alkyl)(C3-C6)cycloalkyl, (C0-C4 alkyl)(C2-C5 heterocyclic), (C0-C4 alkyl)(C6-C10 aryl)R7, and (C1-C4 alkyl)(C3-C9 heteroaryl), wherein R7 is H or OH, wherein optionally the side chain of the amino acid of Formula IV comprises a free amino group,
• (c) an α,α-disubstituted amino acid at position 20,
• (d) up to ten additional amino acid modifications relative to SEQ ID NO:
• 1,
• wherein, when the analog lacks a hydrophilic moiety, the glucagon analog exhibits at least 0.1% activity of native GIP at the GIP receptor, wherein the glucagon analog has less than 100-fold selectivity for the human GLP-1 receptor versus the GIP receptor.

An analog comprising a parent sequence with a total of up to 3 amino acid modifications relative to the parent sequence, wherein the parent sequence is any of SEQ ID NOs:

• 27-33, 35-41, 43-46, 76-80, 83-87, 89, 90, 94-100, 102-112, 120-124, and 127-131, or any of SEQ ID NOs:
• 48, 52, 53, and 74, or any of SEQ ID NOs:
• 50, 51, 54, 56, 58-60, 62-66, 68-70, 72, 73, 75, 81, 82, 88, 92, 114-119, 125, 126, and 133, wherein the amino acid modifications are selected from the group consisting of:
• a DPP-IV protective amino acid at position 2;
• optionally AIB or D-Ser;
• b. a large, aliphatic, nonpolar amino acid at position 12, optionally Ile;
• c. an amino acid orther than Arg at position 17, optionally Gln;
• d. a small aliphatic amino acid at position 18, optionally Ala;
• e. an amino acid other than Asp at position 21, optionally Glu;
• f. an amino acid other than Gln at position 24, optionally Asn or Ala;
• g. an amino acid other than Met at position 27, optionally Leu;
• h. an amino acid other than Asn at position 28, optionally Ala;
• i. an amino acid other than Thr at position 29, optionally Gly;
• j. an extension of 1 to 21 amino acids C-terminal to the amino acid at position 29;
• and
• k. the amino acid sequence set forth as any of SEQ ID NOs:
• 48, 52, 53, and 74.