Glucagon superfamily peptides exhibiting nuclear hormone receptor activity

Patent metadata

Field	Value
Publication number	US-9783592-B2
Application number	US-201514808668-A
Country	US
Kind code	B2
Filing date	Jul 24, 2015
Priority date	May 13, 2010
Publication date	Oct 10, 2017
Grant date	Oct 10, 2017

Abstract

Provided herein are glucagon superfamily peptides conjugated with NHR ligands that are capable of acting at a nuclear hormone receptor. Also provided herein are pharmaceutical compositions and kits of the conjugates of the invention. Further provided herein are methods of treating a disease, e.g., a metabolic disorder, such as diabetes and obesity, comprising administering the conjugates of the invention.

First claim

What is claimed:

1. A compound comprising the structure Q-L-Y;
wherein Q is a glucagon superfamily peptide;
Y is a steroid, which antagonizes a Type I nuclear hormone receptor (NHR) activity, with an EC 50 of about 1μM or less, and has a molecular weight of up to about 1000 daltons; and
L is a linking group or a bond, with the proviso that Y is not a bile salt or cholesterol.

2. The compound of claim 1 , wherein Q exhibits at least 0.1% of the activity of native glucagon-like peptide-1 (GLP-1) at the GLP-1 receptor, at least 0.1% of the activity of native glucagon at the glucagon receptor, or at least 0.1% of the activity of native gastric inhibitory polypeptide (GIP) at the GIP receptor, or a combination thereof.

3. The compound of claim 1 , wherein Q has an EC 50 at the GLP-1 receptor within 10-fold of the EC 50 of Y at the nuclear hormone receptor, an EC 50 at the glucagon receptor within 10-fold of the EC 50 of Y at the nuclear hormone receptor, or an EC 50 at the GIP receptor within 10-fold of the EC 50 of Y at the nuclear hormone receptor, or a combination thereof.

4. The compound of claim 1 , wherein Y acts at a nuclear hormone receptor selected from the group consisting of an estrogen receptor, glucocorticoid receptor, a mineralocorticoid receptor, a progesterone receptor, and an androgen receptor.

5. The compound of claim 4 , wherein L is stable in vivo, hydrolyzable in vivo, or metastable in vivo.

6. The compound of claim 5 , wherein L comprises an ether moiety, an amide moiety, an ester moiety, an acid-labile moiety, a reduction-labile moiety, an enzyme-labile moiety, a hydrazone moiety, a disulfide moiety, or a cathepsin-cleavable moiety.

7. The compound of claim 4 , wherein Q comprises
(I) an amino acid sequence at least 50% identical to native glucagon that retains the alpha-helix conformation of the amino acids corresponding to amino acids 12-29 of native glucagon (SEQ ID NO: 1601);
(II) an amino acid sequence at least 50% identical to native GLP-1 that retains the alpha-helix conformation of the amino acids corresponding to amino acids 12-29 of native GLP-1 (SEQ ID NO: 1603); or
(III) the amino acid sequence:
X 1 -X 2 -Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Ser-Arg-Arg -Ala-Gln-Asp-Phe-Val-Gln-Trp-Leu-Met-Z (SEQ ID NO: 839) with 1 to 3 amino acid modifications thereto,
wherein X 1 and/or X 2 is a non-native amino acid (relative to SEQ ID NO: 1601) that reduces susceptibility of the glucagon peptide to cleavage by dipeptidyl peptidase IV (DPP-IV),
wherein Z is selected from the group consisting of —COOH, -Asn-COOH, Asn -Thr-COOH, and W-COOH, wherein W is 1 to 2 amino acids or GPSSGAPPPS (SEQ ID NO: 1610),
wherein Q comprises a modification selected from the group consisting of:
(i) a lactam bridge between the side chains of amino acids at positions i and i +4, wherein i is 12, 16, 20 or 24, and
(ii) one, two, three, or all of the amino acids at positions 16, 20, 21, and 24 of the glucagon peptide is substituted with an α, α-disubstituted amino acid; and,
wherein Q exhibits glucagon agonist activity.

8. The compound of claim 7 , wherein Q comprises the amino acid sequence SEQ ID NO: 1601 and comprises:
(a) at least one amino acid modification selected from the group consisting of:
(i) substitution of Thr at position 29 with a charged amino acid;
(ii) substitution of Thr at position 29 with a charged amino acid selected from the group consisting of Lys, Arg, His, Asp, Glu, cysteic acid, and homocysteic acid;
(iii) substitution at position 29 with Asp, Glu, or Lys;
(iv) substitution at position 29 with Glu;
(v) insertion after position 29 of 1 to 3 charged amino acids;
(vi) insertion after position 29 of Glu or Lys;
(vii) insertion after position 29 of Gly-Lys, or Lys-Lys;
(viii) substitution of Gln at position 3 with an amino acid comprising a side chain of Structure I, II, or III:

wherein R 1 is C 0-3 alkyl or C 0-3 heteroalkyl; R 2 is NHR 4 or C 1-3 alkyl; R 3 is C 1-3 alkyl; R 4 is H or C 1-3 alkyl; X is NH, O, or S; and Y is NHR 4 , SR 3 , or OR 3 ; and
(ix) a combination thereof; and,
(b) substitution of Ser at position 16 with Thr, Glu, or Aib; and at least one amino acid modification selected from the group consisting of:
(i) substitution of His at position 1 with a non-native amino acid that reduces susceptibility of the glucagon peptide to cleavage by dipeptidyl peptidase IV (DPP-IV),
(ii) substitution of Ser at position 2 with a non-native amino acid that reduces susceptibility of the glucagon peptide to cleavage by dipeptidyl peptidase IV (DPP-IV),
(iii) substitution of Thr at position 7 with Ile, Abu, or Val;
(iv) substitution of Gln at position 20 with Ser, Thr, Ala, Aib, Arg, or Lys;
(v) substitution of Met at position 27 with Leu or Nle;
(vi) deletion of amino acids at positions 28-29;
(vii) deletion of the amino acid at positions 29;
(viii) addition of the amino acid sequence GPSSGAPPPS (SEQ ID NO: 1610) to the C-terminus;
(ix) addition of the amino acid sequence GPSSGAPPPSX (SEQ ID NO: 1450) to the C-terminus, wherein X is any amino acid; and
(x) a combination thereof; and,
wherein Q exhibits glucagon agonist activity.

9. The compound of claim 7 , wherein
(I) Q comprises a glucagon related peptide of SEQ ID NO: 1601, with the following modifications:
(a) an amino acid modification at position 1 that confers GIP agonist activity; and,
(b) a modification selected from the group consisting of:
(i) a lactam bridge between the side chains of amino acids at positions i and i +4 or between the side chains of amino acids at positions j and j +3, wherein i is 12, 13, 16, 17, 20 or 24, and wherein j is 17, and
(ii) one, two, three, or all of the amino acids at positions 16, 20, 21, and 24 of the analog is substituted with an α,α-disubstituted amino acid; and,
(c) 1-10 further amino acid modifications; and,
wherein Q exhibits activity at the GIP receptor; or,
(II) Q comprises the amino acid sequence:
X 1 -X 2 -Gln-Gly-Thr-Phe-Thr-Ser-Asp-X 3 -Ser-X4-Tyr-Leu-X 5 -X 6 -X 7 -X 8 -Ala-X 9 -X 10 -Phe-X 11 -X 12 -Trp-Leu-X 13 -X 14 -X 15 (SEQ ID NO: 55), or an analog thereof, wherein said analog differs from SEQ ID NO: 55 by 1 to 3 amino acid modifications, selected from positions 1, 2, 3, 5, 7, 10, 11, 13, 14, 17, 18, 19, 21, and 25, wherein:
X 1 is His, D-His, (Des-amino)His, hydroxyl-His, acetyl-His, homo-His, or alpha, alpha-dimethyl imidazole acetic acid (DMIA), N-methyl His, alpha-methyl His, or imidazole acetic acid;
X 2 is Ser, D-Ser, Ala, D-Ala, Val, Gly, N-methyl Ser, aminoisobutyric acid (Aib) or N-methyl Ala;
X 3 , X 4 , X 5 , X 10 , X 11 , and X 14 are, individually, any amino acid;
X 6 is Ser, Glu, Gln, homoglutamic acid or homocysteic acid;
X 7 is Arg, Gln, Lys, Cys, Orn, homocysteine or acetyl phenylalanine;
X 8 is Arg, Ala, Lys, Cys, Orn, homocysteine or acetyl phenyalanine;
X 9 is Gln, Lys, Arg, Orn or Citrulline;
X 12 is Ala, Gln, Glu, Lys, Cys, Orn, homocysteine or acetyl phenyalanine;
X 13 is Met, Leu or Nle;
X 15 is Thr, Gly, Lys, Cys, Orn, homocysteine or acetyl phenyalanine; and
wherein Q optionally comprises one of the following modifications:
(a) deletion of amino acids at positions 28-29;
(b) deletion of the amino acid at positions 29;
(c) addition of the amino acid sequence GPSSGAPPPS (SEQ ID NO: 1610) to the C-terminus; and
(d) addition of the amino acid sequence GPSSGAPPPSX (SEQ ID NO: 1450) to the C-terminus, wherein X is any amino acid; and
(e) a co

Assignees

• Univ Indiana Res & Tech Corp

Inventors

• Dimarchi Richard D
• Yang Bin
• Finan Brian

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P3/06

  Antihyperlipidemics · CPC title

• A61P25/28

  for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia · CPC title

• A61P3/00

  Drugs for disorders of the metabolism (of the blood or the extracellular fluid A61P7/00) · CPC title