Methods of treatment using mutant FGF-21 peptide conjugates
Therapeutic regimens and uses of mutant Fibroblast Growth Factor-21 (FGF-21) peptide conjugates comprising a polyethylene glycol (PEG) moiety attached to a mutant FGF-21 peptide via a glycosyl moiety thereof in the treatment of nonalcoholic steatohepatitis are provided.
1. A method of treating nonalcoholic steatohepatitis (NASH) in a subject in need thereof, comprising:
administering once a week to the subject in need thereof a pharmaceutical composition comprising from 10 mg to 50 mg of a mutant Fibroblast Growth Factor-21 (FGF-21) peptide conjugate and a pharmaceutically acceptable carrier,
wherein the mutant FGF-21 peptide conjugate comprises:
i) a mutant FGF-21 peptide comprising the amino acid sequence of SEQ ID NO: 2,
ii) a glycosyl moiety, and
iii) a 20 kDa polyethylene glycol (PEG), wherein the mutant FGF-21 peptide is attached to the glycosyl moiety by a covalent bond between a threonine at amino acid position 173 of SEQ ID NO: 2 and a first site of the glycosyl moiety and wherein the glycosyl moiety is attached to the 20 kDa PEG by a covalent bond between a second site of the glycosyl moiety and the 20 kDa PEG,
wherein administration of the pharmaceutical composition results in at least one of:
a reduction of the liver size as assessed by Magnetic resonance imaging—Proton density fat fraction,
a reduction of the levels of one or more biomarkers comprising Pro-C3, alanine transaminase (ALT), Enhanced LiverFibrosis (ELF) panel, CK-18, inflammation marker high-sensitivity C-reactive protein (hs-CRP), Hemoglobin A1c (HbA1c), Triglycerides, and LDL-c, and
an increase of the levels of HDL-c and/or High Molecular Weight Adiponectin.
2. The method of claim 1 , wherein the subject is a human subject.
3. The method of claim 1 , wherein the pharmaceutical composition is administered sub-subcutaneously.
4. The method of claim 1 , wherein the glycosyl moiety comprises at least one of an N-acetylgalactosamine (GalNAc) residue, a galactose (Gal) residue, a sialic acid (Sia) residue, a 5-amine analogue of a Sia residue, a mannose (Man) residue, mannosamine, a glucose (Glc) residue, an N-acetylglucosamine (GlcNAc) residue, a fucose residue, a xylose residue, or a combination thereof.
5. The method of claim 1 , wherein the glycosyl moiety comprises at least one N-acetylgalactosamine (GalNAc) residue, at least one galactose (Gal) residue, at least one sialic acid (Sia) residue, or a combination thereof.
6. The method of claim 5 , wherein the at least one Sia residue is a nine-carbon carboxylated sugar.
7. The method of claim 6 , wherein the at least one Sia residue is N-acetyl-neuraminic acid (2-keto-5-acetamido-3,5-dideoxy-D-glycero-D-galactononulopyranos-1-onic acid (Neu5Ac), N-glycolylneuraminic acid (Neu5Gc), 2-keto-3-deoxy-nonulosonic acid (KDN), or a 9-substituted sialic acid.
8. The method of claim 7 , wherein the 9-substituted sialic acid is 9-O-lactyl-Neu5 Ac, 9-O-acetyl-Neu5Ac, 9-deoxy-9-fluoro-Neu5Ac, or 9-azido-9-deoxy-Neu5Ac.
9. The method of claim 1 , wherein the glycosyl moiety comprises the structure -GalNAc-Sia-.
10. The method of claim 1 , wherein the 20 kDa PEG moiety is attached to the glycosyl moiety by a covalent bond to a linker, wherein the linker comprises at least one amino acid residue.
11. The method of claim 10 , wherein the at least one amino acid residue is a glycine (Gly).
12. The method of claim 1 , wherein the mutant FGF-21 peptide conjugate comprises the structure -GalNAc-Sia-Gly-PEG (20 kDa).
13. The method of claim 1 , wherein the mutant FGF-21 peptide conjugate comprises the structure:
wherein n is an integer selected from 450 to 460.
14. The method of claim 1 , wherein the 20 kDa PEG is a linear or branched PEG.
15. The method of claim 1 , wherein the 20 kDa PEG is a 20 kDa methoxy-PEG.
16. The method of claim 1 , wherein the mutant FGF-21 peptide conjugate displays an equal or higher potency than wild type FGF-21 when tested in vitro in KLB-FGFR1, KLB-FGFR2 and KLB-FGFR3 expressing cells.
17. The method of claim 1 , comprising administering once a week to the subject in need thereof the pharmaceutical composition comprising about 30 mg of the mutant FGF-21 peptide conjugate.
18. The method of claim 1 , comprising administering once a week to the subject in need thereof the pharmaceutical composition comprising from 25 mg to 30 mg of the mutant FGF-21 peptide conjugate.
19. A method of treating nonalcoholic steatohepatitis (NASH) in a subject in need thereof, comprising:
administering once every two weeks to the subject in need thereof a pharmaceutical composition comprising from 18 mg to 44 mg of a mutant Fibroblast Growth Factor-21 (FGF-21) peptide conjugate and a pharmaceutically acceptable carrier, wherein the mutant FGF-21 peptide conjugate comprises
i) a mutant FGF-21 peptide comprising the amino acid sequence of SEQ ID NO: 2,
ii) a glycosyl moiety, and
iii) a 20 kDa polyethylene glycol (PEG), wherein the mutant FGF-21 peptide is attached to the glycosyl moiety by a covalent bond between a threonine at amino acid position 173 of SEQ ID NO: 2 and a first site of the glycosyl moiety and wherein the glycosyl moiety is attached to the 20 kDa PEG by a covalent bond between a second site of the glycosyl moiety and the 20 kDa PEG,
wherein administration of the pharmaceutical composition results in at least one of:
a reduction of the liver size as assessed by Magnetic resonance imaging —Proton density fat fraction,
a reduction of the levels of one or more biomarkers comprising Pro-C3, alanine transaminase (ALT), Enhanced Liver Fibrosis (ELF) panel, CK-18, inflammation marker high-sensitivity C-reactive protein (hs-CRP), Hemoglobin A1c (HbA1c), Triglycerides, and LDL-c, and
an increase of the levels of HDL-c and/or High Molecular Weight Adiponectin.
20. The method of claim 19 , wherein the subject is a human subject.
21. The method of claim 19 , wherein the pharmaceutical composition is administered sub-subcutaneously.
22. The method of claim 19 , wherein the glycosyl moiety comprises at least one of an N-acetylgalactosamine (GalNAc) residue, a galactose (Gal) residue, a sialic acid (Sia) residue, a 5-amine analogue of a Sia residue, a mannose (Man) residue, mannosamine, a glucose (Glc) residue, an N-acetylglucosamine (GlcNAc) residue, a fucose residue, a xylose residue, or a combination thereof.
23. The method of claim 19 , wherein the glycosyl moiety comprises at least one N-acetylgalactosamine (GalNAc) residue, at least one galactose (Gal) residue, at least one sialic acid (Sia) residue, or a combination thereof.
24. The method of claim 23 , wherein the at least one Sia residue is a nine-carbon carboxylated sugar.
25. The method of claim 24 , wherein the at least one Sia residue is N-acetyl-neuraminic acid (2-keto-5-acetamido-3,5-dideoxy-D-glycero-D-galactononulopyranos-1-onic acid (Neu5Ac), N-glycolylneuraminic acid (Neu5Gc), 2-keto-3-deoxy-nonulosonic acid (KDN), or a 9-substituted sialic acid.
26. The method of claim 25 , wherein the 9-substituted sialic acid is 9-O-lactyl-Neu5Ac, 9-O-acetyl-Neu5Ac, 9-deoxy-9-fluoro-Neu5Ac, or 9-azido-9-deoxy-Neu5Ac.
27. The method of claim 19 , wherein the glycosyl moiety comprises the structure -GalNAc-Sia-.
28. The method of claim 19 , wherein the 20 kDa PEG moiety is attached to the glycosyl moiety by a covalent bond to a linker, wherein the linker comprises at least one amino acid residue.
29. The method of claim 28 , wherein the at least one amino acid residue is a glycine (Gly).
30. The method of claim 19 , wherein the mutant FGF-21 peptide conjugate comprises the structure -GalNAc-Sia-Gly-PEG (20 kDa).
31. The method of claim 19 , wherein the mutant FGF-21 peptide conjugate comprises the structure:
wherein n is an integer selected from 450 to 460.
32. The method of claim 19 , wherein the 20 kDa PEG is a linear or branched PEG.
33. The method of claim 19 , wherein the 20 kDa PEG is a 20 kDa methoxy-PEG.
34. The method of claim 19 , wherein the mutant FGF-21 peptide conjugate displays an equal or higher potency than wild type FGF-21 when tested in vitro in KLB-FGFR1, KLB-FGFR2 and KLB-FGFR3 expressing cells.
35. The method of claim 18 , comprising administering once every two weeks to the subject in need thereof the pharmaceutical composition comprising about 40 mg of the mutant FGF-21 peptide conjugate.
36. The method of claim 19 , comprising administering once every two weeks to the subject in need thereof the pharmaceutical composition comprising from 36 mg to 44 mg of the mutant FGF-21 peptide conjugate.