# US 11,993,637 B2 · Modified FGF-21 polypeptides with non-naturally encoded amino acids

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- Number: US 11,993,637 B2
- Document ID: US11993637B2
- Status: Granted
- Kind: B2
- Application number: 17/165,351
- Filed: 2021-02-02
- Published: 2021-08-26
- Granted: 2024-05-28
- Earliest claimed: 2007-03-30
- Assignees: Ambrx, Inc.
- Applicants: Ambrx, Inc.
- Inventors: Thomas P. Cujec; Roberto Mariani; Anna-Maria A. Hays Putnam; William M. Keefe; Nick Knudsen; Lillian Skidmore; Jason Pinkstaff; Vadim Kraynov
- Web page: https://patentgrep.com/patent/US11993637B2

## CPC

- C07K 14/50: Fibroblast growth factor [FGF]
- A61K 38/1825: {Fibroblast growth factor [FGF]}
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61K 47/60: the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
- A61P 1/18: for pancreatic disorders, e.g. pancreatic enzymes
- A61P 3/00: Drugs for disorders of the metabolism (of the blood or the extracellular fluid A61P7/00)
- A61P 3/04: Anorexiants; Antiobesity agents
- A61P 3/08: for glucose homeostasis (pancreatic hormones A61P5/48)
- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- A61P 9/10: for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
- A61P 11/00: Drugs for disorders of the respiratory system
- A61P 17/02: for treating wounds, ulcers, burns, scars, keloids, or the like
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 31/04: Antibacterial agents
- A61P 37/02: Immunomodulators
- A61P 39/00: General protective or antinoxious agents
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C12N 15/70: Vectors or expression systems specially adapted for E. coli
- 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)
- 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
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

Modified FGF-21 polypeptides and uses thereof are provided.

## Claims

### Claim 1

A modified FGF-21 polypeptide comprising a non-naturally encoded amino acid,
wherein said modified FGF-21 polypeptide maintains the biological activity of human FGF-21 and comprises a non-naturally encoded amino acid, and:
(a) the modified FGF-21 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:
wherein the R group is any substituent other than the side chain found in alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, pyrrolysine, or selenocysteine;
(c) the modified FGF-21 polypeptide contains a substitution of an amino acid with the non-naturally encoded amino acid at a position in said modified FGF-21 polypeptide corresponding to residue 72, 77, 86, 108, 110, 131, or 146 of SEQ ID NO: 1;
(d) the non-naturally encoded amino acid is linked to a linker or a polymer; and
(e) the FGF-21 polypeptide has an in vivo half-life which is at least two-fold greater than the human FGF-21 polypeptide in SEQ ID NO: 1.

### Claim 2 (depends on 1)

The modified FGF-21 polypeptide of claim 1, wherein the non-naturally encoded amino acid is directly or indirectly linked to a polymer.

### Claim 3 (depends on 1)

The modified FGF-21 polypeptide of claim 1, wherein the non-naturally encoded amino acid is directly or indirectly linked to a water soluble polymer.

### Claim 4 (depends on 3)

The modified FGF-21 polypeptide of claim 3, wherein the water soluble polymer comprises a poly(ethylene glycol) moiety.

### Claim 5 (depends on 1)

The modified FGF-21 polypeptide of claim 1, wherein the non-naturally encoded amino acid is at position 108, of the FGF-21 polypeptide of SEQ ID NO: 1 or the corresponding position in any one of the FGF-21 polypeptides of SEQ ID NOs: 2-7.

### Claim 6 (depends on 3)

The modified FGF-21 polypeptide of claim 3, wherein the water soluble polymer has a molecular weight of between about 0.1 kDa and about 100 kDa.

### Claim 7 (depends on 3)

The modified FGF-21 polypeptide of claim 3, which is made by reacting the non-naturally encoded amino acid of the modified FGF-21 polypeptide with a water soluble polymer comprising an aminooxy, hydrazine, hydrazide or semicarbazide group.

### Claim 8 (depends on 7)

The modified FGF-21 polypeptide of claim 7, wherein the aminooxy, hydrazine, hydrazide or semicarbazide group of the water soluble polymer is linked to the non-naturally encoded amino acid of the modified FGF-21 polypeptide through an amide linkage.

### Claim 9 (depends on 1)

The modified FGF-21 polypeptide of claim 1, wherein the FGF-21 polypeptide comprises the FGF-21 polypeptide of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO: 7, and said non-naturally encoded amino acid is at a position selected from residue 72, 77, 86, 108, 110, 131, and 146 of SEQ ID NO: 1, or the corresponding position of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO: 7.

### Claim 10 (depends on 1)

The modified FGF-21 polypeptide of claim 1, wherein the FGF-21 polypeptide comprises the FGF-21 polypeptide of SEQ ID NO: 1, and said non-naturally encoded amino acid is at a position selected from residue 72, 77, 86, 108, 110, 131, and 146 of SEQ ID NO: 1.

### Claim 11 (depends on 9)

The modified FGF-21 polypeptide of claim 9, wherein the non-naturally encoded amino acid is directly or indirectly linked to a polymer.

### Claim 12 (depends on 10)

The modified FGF-21 polypeptide of claim 10, wherein the non-naturally encoded amino acid is directly or indirectly linked to a polymer.

### Claim 13 (depends on 11)

The modified FGF-21 polypeptide of claim 11, wherein the polymer comprises a poly(ethylene glycol) moiety.

### Claim 14 (depends on 12)

The modified FGF-21 polypeptide of claim 12, wherein polymer comprises a poly(ethylene glycol) moiety.

### Claim 15 (depends on 11)

The modified FGF-21 polypeptide of claim 11, wherein the polymer comprises a poly(ethylene glycol) moiety which has a molecular weight of between about 0.1 kDa and about 100 kDa.

### Claim 16 (depends on 12)

The modified FGF-21 polypeptide of claim 12, wherein polymer comprises a poly(ethylene glycol) moiety which has a molecular weight of between about 0.1 kDa and about 100 kDa.

### Claim 17 (depends on 1)

A composition comprising a modified FGF-21 polypeptide of claim 1, and a pharmaceutically acceptable carrier.

### Claim 18 (depends on 9)

A composition comprising a modified FGF-21 polypeptide of claim 9, and a pharmaceutically acceptable carrier.

### Claim 19 (depends on 10)

A composition comprising a modified FGF-21 polypeptide of claim 10, and a pharmaceutically acceptable carrier.

### Claim 20 (depends on 13)

A composition comprising a modified FGF-21 polypeptide of claim 13, and a pharmaceutically acceptable carrier.