Methods of treatment using modified FGF-21 polypeptides comprising non-naturally occurring amino acids

• US 9,517,273 B2
• Filed: 04/07/2015
• Issued: 12/13/2016
• Est. Priority Date: 03/30/2007
• Status: Active Grant

• Current Assignee: Ambrx, Inc. (Johnson & Johnson)
• Original Assignee: Ambrx, Inc. (Johnson & Johnson)
• Inventors: Keefe, Wiliam M., Cujec, Thomas P., Mariani, Roberto, Hays Putnam, Anna-Maria A., Knudsen, Nick, Skidmore, Lillian, Pinkstaff, Jason, Kraynov, Vadim
• Primary Examiner(s): STOICA, ELLY GERALD
• Application Number:
  US14/680,543
• Publication Number:
  US 20150273075A1
• Time in Patent Office: 616 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/1825 Fibroblast growth factor [FGF]
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/08 for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 39/00 General protective or antinoxious agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/50 Fibroblast growth factor [FGF]
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

Modified FGF-21 polypeptides and uses thereof are provided.

Claims

1. A method of treating a patient in need thereof, comprising administering an effective amount of a modified FGF-21 polypeptide to said patient, wherein said modified FGF-21 polypeptide maintains the biological activity of human FGF-21 polypeptides and comprises a non-naturally encoded amino acid, wherein:

(a) the polypeptide comprises a sequence at least 90% identical to SEQ ID NO;

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, or SEQ ID NO;

7;

(b) the non-naturally encoded amino acid has the structure;

2. The method of claim 1, wherein the non-naturally encoded amino acid is a phenylalanine derivative.

3. The method of claim 1, wherein the non-naturally encoded amino acid is linked to a polymer.

4. The method of claim 1, wherein the non-naturally encoded amino acid is selected from a para-substituted, ortho-substituted, or meta-substituted phenylalanine comprising a carbonyl group, an aminooxy group, a hydrazide group, a hydrazine group, a semicarbazide group, an azide group, or an alkyne group.

5. The method of claim 1, wherein said patient has a disease selected from type 2 diabetes, obesity, insulin resistance, hyperinsulinemia, glucose intolerance, hyperglycemia, metabolic syndrome, a glucose metabolism disorder, or a combination thereof.

6. A method of treating a patient in need thereof, comprising administering an effective amount of a modified FGF-21 polypeptide to said patient, wherein said modified FGF-21 polypeptide maintains the biological activity of human FGF-21 polypeptides and comprises the polypeptide of SEQ ID NO:

1 except that an amino acid in the polypeptide is substituted by a non-naturally encoded amino acid, wherein;

(a) said non-naturally encoded amino acid is at a position corresponding to residue 108 of SEQ ID NO;

1;

(b) said non-naturally encoded amino acid comprises para-acetyl phenylalanine linked to a polymer comprising a poly(ethylene glycol), wherein said poly(ethylene glycol) has an average molecular weight of about 30 kDa; and

(c) said non-naturally encoded amino acid is linked to said polymer through an oxime linkage.

7. The method of claim 6, wherein said polymer comprises an aminooxy group, and said non-naturally encoded amino acid is linked to said polymer by said oxime linkage formed by a reaction of said para-acetyl phenylalanine with said aminooxy group.

8. A method of treating a patient in need thereof, comprising administering an effective amount of a modified FGF-21 polypeptide to said patient, wherein said modified FGF-21 polypeptide maintains the biological activity of human FGF-21 polypeptides and comprises a non-naturally encoded amino acid, wherein:

(a) the polypeptide comprises a sequence at least 95% identical to SEQ ID NO;

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, or SEQ ID NO;

7;

(b) the non-naturally encoded amino acid has the structure;

9. The method of claim 8, wherein the non-naturally encoded amino acid is a phenylalanine derivative or comprises a first functional group and the linker, polymer, or biologically active molecule comprises a second functional group, wherein the first functional group and second functional group are not identical and each comprise a carbonyl group, an aminooxy group, a hydrazide group, a hydrazine group, a semicarbazide group, an azide group, or an alkyne group.

10. The method of claim 9, wherein the first functional group on the non-naturally encoded amino acid is a carbonyl moiety and the second functional group on the linker, polymer, or biologically active molecule is an aminooxy moiety, and the resultant covalent linkage created by the reaction of the first and second functional groups is an oxime linkage.

11. The method of claim 8, wherein the non-naturally encoded amino acid is linked to a polymer comprising poly(ethylene glycol) or is linked to a polymer comprising poly(ethylene glycol) having an average molecular weight of between about 0.1 kDa and about 100 kDa.

12. The method of claim 11, wherein the poly(ethylene glycol) has an average molecular weight of about 30 kDa.

13. The method of claim 8, wherein the polypeptide has an in vivo half-life at least five-fold greater than the human FGF-21 polypeptide in SEQ ID NO:

1.

14. The method of claim 8, wherein the non-naturally encoded amino acid is at a position corresponding to residue 108 of SEQ ID NO:

1.

15. The method of claim 8, wherein the non-naturally encoded amino acid is selected from a para-substituted, ortho-substituted, or meta-substituted phenylalanine comprising a carbonyl group, an aminooxy group, a hydrazide group, a hydrazine group, a semicarbazide group, an azide group, or an alkyne group.

16. The method of claim 15, wherein the non-naturally encoded amino acid is para-acetyl-L-phenylalanine.

17. The method of claim 8, wherein the polypeptide comprises SEQ ID NO:

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, or SEQ ID NO;

7, containing a substitution of an amino acid with a non-naturally encoded amino acid.

18. The method of claim 8, wherein said patient has a disease selected from type 2 diabetes, obesity, insulin resistance, hyperinsulinemia, glucose intolerance, hyperglycemia, metabolic syndrome, a glucose metabolism disorder, or a combination thereof.

19. The method of claim 8, further comprising administering another active agent comprising anti-diabetic agent to the patient.

20. The method of claim 8, further comprising administering to the patient another active agent selected from insulin, biguanides, thiazolidinediones, sulfonylureas, benzoic acid derivatives, glucosidase inhibitors, DPP-4 inhibitors, human insulin, zinc insulin, protamine zinc insulin, a fragment or derivative of insulin, INS-1, oral insulin, insulin sensitizers, pioglitazone or a salt thereof, pioglitazone hydrochloride, rosiglitazone or a salt thereof, rosiglitazone maleate, netoglitazone, Rivoglitazone (CS-011), FK-614, Tesaglitazar (AZ-242), Ragaglitazar (N,N-622), Muraglitazar (BMS-298585), Edaglitazone (BM-13-1258), Metaglidasen (MBX-102), Naveglitazar (LY-519818), MX-6054, LY-510929, AMG-131(T-131), THR-0921, alpha-glucosidase inhibitors, voglibose, acarbose, miglitol, emiglitate, phenformin, metformin, metformin HCl, buformin or a salt thereof, buformin hydrochloride, buformin fumarate, buformin succinate, insulin secretagogues, sulfonylurea, tolbutamide, glyburide, micronized glyburide, glimepiride, glibenclamide, gliclazide, chlorpropamide, tolazamide, acetohexamide, glyclopyramide, glimepiride, AG-EE 623 Z, glipizide, glipizide GITS, glybuzole, meglitinides, repaglinide, nateglinide, mitiglinide, mitiglinide calcium salt hydrate, dipeptidyl peptidase IV inhibitors, Vidagliptin (LAF237), P32/98, Sitagliptin (MK-431), P93/01, PT-100, Saxagliptin (BMS-477118), T-6666, TS-021, beta3 agonists, AJ-9677, GPR40 agonists, glucagon-like polypeptides (I) (glp I), glp2, diabetogenic peptide hormones, GLP-1 receptor agonists, GLP-1, GLP-1MR agent, N,N-2211, AC-2993 (exendin-4), BIM-51077, Aib(8,35)hGLP-1 (7,37)NH2, GLP-1(7-36)amide, CJC-[131], amylin agonists, pramlintide, phosphotyrosine phosphatase inhibitors, sodium vanadate, gluconeogenesis inhibitors, glycogen phosphorylase inhibitors, glucose-6-phosphatase inhibitors, glucagon antagonists, SGLUT (sodium-glucose cotransporter) inhibitors, T-1095, 11beta-hydroxysteroid dehydrogenase inhibitors, BVT-3498, adiponectin, adiponectin agonists, IKK inhibitors, AS-2868, leptin resistance improving drugs, somatostatin receptor agonists, glucokinase activators, Ro-28-1675, Glucose-dependent insulinotropic peptide (GIP), Vildagliptin (Galvus) (LAF237), Sitagliptin (Januvia), Saxagliptin, Alogliptin, 1-[[[2-[(5-cyanopyridin-2-yl)amino]ethyl]amino]acetyl]-2-cyano-(- S)-pyrrolidine (NVP DPP728), troglitazone, carnitine palmitoyl-transferase I inhibitors, and etomoxir.