Engineered primate cystine/cysteine degrading enzymes as antineogenic agents

Patent metadata

Field	Value
Publication number	US-10363311-B2
Application number	US-201414472779-A
Country	US
Kind code	B2
Filing date	Aug 29, 2014
Priority date	Aug 29, 2013
Publication date	Jul 30, 2019
Grant date	Jul 30, 2019

Abstract

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

First claim

What is claimed is:

1. An isolated, modified primate cystathionine-γ-lyase (CGL) enzyme having at least one substitution relative to a native primate CGL amino acid sequence (see SEQ ID NOs: 1 and 7-10), said at least one substitution including a threonine at position 59 of the native primate CGL sequence.

2. The enzyme of claim 1 , further comprising a valine substitution at position 339.

3. The enzyme of claim 1 , further comprising a heterologous peptide segment.

4. The enzyme of claim 3 , wherein the heterologous peptide segment is an XTEN peptide, an IgG Fc, an albumin, or an albumin binding peptide.

5. The enzyme of claim 1 , wherein the enzyme is coupled to polyethylene glycol (PEG).

6. The enzyme of claim 5 , wherein the enzyme is coupled to PEG via one or more lysine or cystine residues.

7. An isolated, modified primate cystathionine-γ-lyase (CGL) enzyme having at least two substitutions relative to a native primate CGL amino acid sequence (see SEQ ID NOs: 1 and 7-10), said at least two substitutions including a threonine at position 59 and a valine at position 339 of the native primate CGL sequence.

8. A nucleic acid comprising a nucleotide sequence encoding the enzyme of claim 1 .

9. The nucleic acid of claim 8 , wherein the nucleic acid is codon optimized for expression in bacteria, fungus, insects, or mammals.

10. An expression vector comprising the nucleic acid of claim 8 .

11. A host cell comprising the nucleic acid of claim 8 .

12. The host cell of claim 11 , wherein the host cell is a bacterial cell, a fungal cell, an insect cell, or a mammalian cell.

Assignees

• Univ Texas

Inventors

• Georgiou George
• Stone Everett

Classifications

• A61P43/00

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

• A61P37/00

  Drugs for immunological or allergic disorders · CPC title

• A61P5/00

  Drugs for disorders of the endocrine system · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• C12Y404/01001

  Cystathionine gamma-lyase (4.4.1.1) · CPC title