Engineered primate cystine/cysteine degrading enzymes as antineogenic agents

Patent metadata

Field	Value
Publication number	US-10716856-B2
Application number	US-201916440222-A
Country	US
Kind code	B2
Filing date	Jun 13, 2019
Priority date	Aug 29, 2013
Publication date	Jul 21, 2020
Grant date	Jul 21, 2020

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. A polypeptide comprising a variant of a native primate cystathionine-gamma-lyase (CGL) enzyme, wherein the variant CGL has cysteine/cystine degrading activity, comprises a sequence at least 95% identical to SEQ ID NO: 1, and comprises E59T and E339V substitutions relative to SEQ ID NO: 1.

2. The polypeptide of claim 1 , further comprising a heterologous peptide segment.

3. The polypeptide of claim 2 , wherein the heterologous peptide segment is an XTEN peptide, an IgG Fc, an albumin, or an albumin binding peptide.

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

5. The polypeptide of claim 4 , wherein the polypeptide is coupled to PEG via one or more lysine or cystine residues.

6. A nucleic acid comprising a nucleotide sequence encoding the polypeptide of claim 1 .

7. The nucleic acid of claim 6 , wherein the nucleic acid is codon optimized for expression in bacteria, fungus, insects, or mammals.

8. An expression vector comprising the nucleic acid of claim 6 .

9. A host cell comprising the nucleic acid of claim 6 .

10. The host cell of claim 9 , wherein the host cell is a bacterial cell, a fungal cell, an insect cell, or a mammalian cell.

11. A pharmaceutical formulation comprising the polypeptide of claim 1 in a pharmaceutically acceptable carrier.

Assignees

• Univ Texas

Inventors

• Georgiou George
• Stone Everett

Classifications

• C12Y404/01001

  Cystathionine gamma-lyase (4.4.1.1) · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• A61K38/51 Primary

  Lyases (4) · CPC title

• A61K45/06 Primary

  Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca · CPC title

• A61P5/00

  Drugs for disorders of the endocrine system · CPC title