Antibody peptide conjugates that have agonist activity at both the glucagon and glucagon-like peptide 1 receptors
Described are antibody peptide conjugates (APCs) comprising an antibody conjugated to a peptide analog of glucagon, which have been modified to be resistant to cleavage and inactivation by dipeptidyl peptidase IV (DPP-IV) and to increase in vivo half-life of the peptide analog while enabling the peptide analog to have agonist activity at the glucagon (GCG) receptor and the glucagon-like peptide 1 (GLP-1) receptor and the use of such APCs for treatment of metabolic disorders such as diabetes, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), and obesity.
1. An antibody peptide conjugate comprising an antibody having a light chain having the amino acid sequence of SEQ ID NO:64 and a heavy chain having the amino acid sequence of SEQ ID NO:65 conjugated to a peptide having the amino acid sequence
(SEQ ID NO: 63)
His-Xaa 2 -Gln-Gly-Thr-Phe-Thr-Ser-Asp-
Xaa 10 -Ser-Xaa 12 -Tyr-Leu-Asp-Xaa 16 -Arg-
Ala-Ala-Xaa 20 -Asp-Phe-Val-Xaa 24 -Xaa 25 -
Xaa 26 -Xaa 27 -Xaa 28 -Xaa 29 -Lys-Xaa 31
wherein
Xaa 2 is α-aminoisobutyric acid (aib) or D-Ser;
Xaa 16 is aib;
Xaa 31 is Lys;
Xaa 10 is Tyr;
X 12 is Lys;
Xaa 20 is Gln;
Xaa 24 is Gln;
Xaa 25 is Trp;
Xaa 26 is Leu;
Xaa 27 is Met;
Xaa 28 is Asp;
Xaa 29 is Thr;
wherein Lys31 is covalently linked at its epsilon amino group to an aminooxy acid residue;
wherein either the light chain includes a substitution of the amino acid at position 125 or 142 with a para-acetylphenylalanine (pAcF) or the heavy chain includes a substitution of the amino acid a position 16, 32, 33, 56, 114, 179, 198, or 211 with a AcF; and
wherein the aminooxy acid residue of the peptide is covelantly linked to the pAcF residue of the antibody.
2. The antibody peptide conjugate of claim 1 , wherein the peptide comprises Lys31 covalently linked at the epsilon amino group directly to the aminooxy acid residue or to the aminooxy acid residue via a polyethylene glycol spacer, γGlu spacer, or γGlu-γGlu spacer.
3. The antibody peptide conjugate of claim 2 , wherein the polyethylene glycol spacer comprises 2, 4, 6, 8, 24, or 36 ethoxy units.
4. A pharmaceutical composition comprising the antibody peptide conjugate of claim 1 and a pharmaceutically acceptable carrier.
5. A method for treating a metabolic disease or disorder in a patient, comprising administering to a patient in need the pharmaceutical composition of claim 4 to treat the metabolic disease or disorder.
6. The method of claim 5 , wherein the metabolic disease or disorder comprises diabetes, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), or obesity.
7. The method of claim 6 , wherein the diabetes comprises Type 1 diabetes, Type II diabetes, or gestational diabetes.