Engineered phenylalanine ammonia lyase polypeptides

Patent metadata

Field	Value
Publication number	US-10487319-B2
Application number	US-201715720060-A
Country	US
Kind code	B2
Filing date	Sep 29, 2017
Priority date	Apr 18, 2013
Publication date	Nov 26, 2019
Grant date	Nov 26, 2019

Abstract

The present invention provides engineered phenylalanine ammonia-lyase (PAL) polypeptides and compositions thereof, as well as polynucleotides encoding the engineered phenylalanine ammonia-lyase (PAL) polypeptides.

First claim

What is claimed is:

1. An engineered polypeptide comprising an amino acid sequence having at least 90% sequence identity to reference sequence SEQ ID NO:4, wherein said amino acid sequence comprises an amino acid residue selected from leucine, methionine, and glutamine at position 305, and further comprising at least one or more of the following substitutions A39V, T54K, G59R, S73K, A112C, R134Q, A91V, Y158H, S180A, K195E, Q240R/W, T243I/L, I245L, A256G, L257W/A, N270K, N290G, Y304H, H307G/Q/M, E3080, I326F, L349M, D353A/N, L364Q, A394V, S399N, N400K, P404A, L407V, F443H, N453G, Y459F, T460G, T463N, N474Q, E509L, Q521K/S, K522Y/F/N, T524S, P528L, S546R, and P564 G/L/M, wherein said positions in said amino acid sequence are numbered in reference to SEQ ID NO:4.

2. The engineered polypeptide of claim 1 , wherein said engineered polypeptide exhibits an improved property selected from reduced sensitivity to proteolysis, increased tolerance to acidic pH, reduced immunogenicity, or a combination thereof, as compared to the reference sequence SEQ ID NO:4.

3. The engineered polypeptide of claim 2 , wherein the improved property is selected from reduced sensitivity to proteolysis and/or increased tolerance to acidic pH.

4. The engineered polypeptide of claim 1 , wherein said engineered polypeptide is resistant to proteolysis, acid stable, and/or deimmunized.

5. The engineered polypeptide of claim 4 , wherein said engineered polypeptide is resistant to proteolysis by at least one digestive tract enzyme, wherein said engineered polypeptide is resistant to proteolysis by chymotrypsin, trypsin, carboxypeptidases, and/or elastases.

6. The engineered polypeptide of claim 4 , wherein said engineered polypeptide is deimmunized.

7. The engineered polypeptide of claim 1 , wherein said polypeptide is purified.

8. A composition comprising at least one engineered polypeptide of claim 1 .

9. The composition of claim 8 , wherein said composition is a pharmaceutical composition.

10. The pharmaceutical composition of claim 9 , wherein said composition is suitable for the treatment of phenylketonuria.

11. The pharmaceutical composition of claim 9 , wherein said composition is suitable for oral administration to a human.

12. The pharmaceutical composition of claim 11 , wherein said composition is in the form of a pill, tablet, capsule, gelcap, liquid, or emulsion.

13. The pharmaceutical composition of claim 12 , wherein said pill, tablet, capsule, or gelcap further comprises an enteric coating.

14. The pharmaceutical composition of claim 9 , wherein said composition is suitable for parenteral injection into a human.

15. The pharmaceutical composition of claim 9 , wherein said composition is coadministered with at least one additional therapeutically effective compound.

Assignees

• Codexis Inc

Inventors

• Huisman Gjalt W
• Agard Nicholas J
• Mijts Benjamin
• Vroom Jonathan
• Zhang Xiyun

Classifications

• A61P43/00

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

• A61P3/00

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

• A61P25/00

  Drugs for disorders of the nervous system · CPC title

• C12N9/88 Primary

  Lyases (4.) · CPC title

• A61K38/00

  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 title