Engineered primate L-methioninase for therapeutic purposes

Patent metadata

Field	Value
Publication number	US-9624484-B2
Application number	US-201615297102-A
Country	US
Kind code	B2
Filing date	Oct 18, 2016
Priority date	Aug 29, 2013
Publication date	Apr 18, 2017
Grant date	Apr 18, 2017

Abstract

Methods and compositions relating to the engineering of an improved protein with methionine-γ-lyase enzyme activity are described. For example, in certain aspects there may be disclosed a modified cystathionine-γ-lyase (CGL) comprising one or more amino acid substitutions and capable of degrading methionine. Furthermore, certain aspects of the invention provide compositions and methods for the treatment of cancer with methionine depletion using the disclosed proteins or nucleic acids.

First claim

What is claimed is:

1. A nucleic acid comprising a nucleotide sequence encoding a modified human cystathionine-γ-lyase (CGL) enzyme having at least the following substitutions relative to a native human CGL amino acid sequence (see SEQ ID NO: 1), said substitutions selected from (a) E59N, S63L, L91M, R119L, K268R, T311G, E339V and I353S; (b) E59I, S63L, L91M, R119L, K268R, T311G, E339V and I353S; (c) E59N, S63L, L91M, R119A, K268R, T311G, E339V and I353S; and (d) E59I, S63L, L91M, R119A, K268R, T311G, E339V and I353S, wherein the modified human CGL enzyme has higher methionine-γ-lyase activity compared to hMGL-NLV.

2. The nucleic acid of claim 1 , wherein the nucleic acid is codon optimized for expression in bacteria, fungus, insects, or mammals.

3. An expression vector comprising the nucleic acid of claim 2 .

4. A host cell comprising the nucleic acid of claim 2 .

5. The host cell of claim 4 , wherein the host cell is a bacterial cell, a fungal cell, an insect cell, or a mammalian cell.

6. The host cell of claim 5 , wherein the bacterial cell is an E. coli strain having deletions of genes ilvA and metA.

7. The nucleic acid of claim 1 , wherein the encoded enzyme has at least the substitutions E59N, S63L, L91M, R119L, K268R, T311G, E339V and I353S.

8. The nucleic acid of claim 1 , wherein the encoded enzyme has at least the substitutions E59I, S63L, L91M, R119L, K268R, T311G, E339V and I353 S.

9. The nucleic acid of claim 1 , wherein the encoded enzyme has at least the substitutions E59N, S63L, L91M, R119A, K268R, T311G, E339V and I353S.

10. The nucleic acid of claim 1 , wherein the encoded enzyme has at least the substitutions E59I, S63L, L91M, R119A, K268R, T311G, E339V and I353S.

11. The nucleic acid of claim 1 , wherein the encoded enzyme further comprises a heterologous peptide segment.

12. The nucleic acid of claim 1 , wherein the heterologous peptide segment is an XTEN peptide, an IgG Fc, an albumin, or an albumin binding peptide.

Assignees

• Univ Texas

Inventors

• Georgiou George
• Stone Everett
• Lu Wei-Cheng

Classifications

• A61P43/00

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

• A61P35/00

  Antineoplastic agents · CPC title

• A61P37/04

  Immunostimulants · CPC title

• C12N1/20

  Bacteria; Culture media therefor · CPC title

• C12N1/14

  Fungi (culture of mushrooms A01G18/00; as new plants A01H15/00); Culture media therefor · CPC title