Human kynureninase enzymes and uses thereof

Patent metadata

Field	Value
Publication number	US-11648272-B2
Application number	US-201916385562-A
Country	US
Kind code	B2
Filing date	Apr 16, 2019
Priority date	Apr 16, 2018
Publication date	May 16, 2023
Grant date	May 16, 2023

Abstract

Methods and compositions related to the use of a protein with kynureninase activity are described. For example, in certain aspects there may be disclosed a modified kynureninase capable of degrading kynurenine. Furthermore, certain aspects of the invention provide compositions and methods for the treatment of cancer with kynurenine depletion using the disclosed proteins or nucleic acids.

First claim

What is claimed is:

1. An isolated, modified kynureninase enzyme having at least 90% sequence identity to a human kynureninase enzyme comprising the amino acid sequence of SEQ ID NO: 1 and comprising at least one substitution selected from the group consisting of A99R, E103R, V104H, R107P, G112Y, I183P, and I331S, wherein the isolated, modified kynureninase enzyme has increased catalytic activity (kcat/kM) towards kynurenine as compared to the human kynureninase enzyme comprising the amino acid sequence of SEQ ID NO: 1.

2. The isolated, modified kynureninase enzyme of claim 1 , comprising:
(a) the A99R substitution;
(b) the G112Y substitution;
(c) the E103R substitution;
(d) the V104H substitution;
(e) the I183P substitution; and/or
(f) the R107P substitution.

3. The isolated, modified kynureninase enzyme of claim 1 , further comprising one or more substitutions selected from the group consisting of L72N, H102W, F249W, A282P, F306W, S408N, N333T, and A436T.

4. The isolated, modified kynureninase enzyme of claim 1 , wherein the isolated, modified kynureninase enzyme has a catalytic activity for kynurenine (KYN) (kcat/kM) of 8000M −1 s −1 to 40000 M −1 s −1 .

5. The isolated, modified kynureninase enzyme of claim 1 , wherein the isolated, modified kynureninase comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 2.

6. The isolated, modified kynureninase enzyme of claim 5 , wherein the isolated, modified kynureninase comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 2.

7. The isolated, modified kynureninase enzyme of claim 1 , further comprising a heterologous peptide segment.

8. The isolated, modified kynureninase enzyme of claim 1 , wherein the isolated, modified kynureninase enzyme is coupled to polyethylene glycol.

9. A pharmaceutical formulation comprising the isolated, modified kynureninase enzyme of claim 1 in a pharmaceutically acceptable carrier.

10. The isolated, modified kynureninase enzyme of claim 1 , wherein the isolated, modified kynureninase comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 3.

11. The isolated, modified kynureninase enzyme of claim 10 , wherein the isolated, modified kynureninase comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 3.

12. An isolated, modified human kynureninase enzyme comprising the amino acid sequence of SEQ ID NO: 3.

13. The isolated, modified kynureninase enzyme of claim 12 , wherein the isolated, modified kynureninase enzyme is coupled to polyethylene glycol.

14. A pharmaceutical formulation comprising the isolated, modified kynureninase enzyme of claim 3 in a pharmaceutically acceptable carrier.

15. A pharmaceutical formulation comprising the isolated, modified kynureninase enzyme of claim 6 in a pharmaceutically acceptable carrier.

16. A pharmaceutical formulation comprising the isolated, modified kynureninase enzyme of claim 8 in a pharmaceutically acceptable carrier.

17. A pharmaceutical formulation comprising the isolated, modified kynureninase enzyme of claim 11 in a pharmaceutically acceptable carrier.

18. A pharmaceutical formulation comprising the isolated, modified kynureninase enzyme of claim 12 in a pharmaceutically acceptable carrier.

19. A pharmaceutical formulation comprising the isolated, modified kynureninase enzyme of claim 13 in a pharmaceutically acceptable carrier.

20. The isolated, modified kynureninase enzyme of claim 1 , wherein the isolated, modified kynureninase enzyme comprises the following substitutions: L72N, A99R, H102W, E103R, V104H, R107P, G112Y, I183P, A282P, F306W, I331S, N333T, S408N, and A436T.

Assignees

• Univ Texas

Inventors

• Georgiou George
• Stone Everett
• Blazeck John
• Karamitros Christos

Classifications

• A61K35/17 Primary

  Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes (when activated by a specific antigen A61K39/00) · CPC title

• C12N9/14 Primary

  Hydrolases (3) · CPC title

• A61K47/60 Primary

  the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol · CPC title

• C07K2319/00

  Fusion polypeptide · CPC title

• A61K38/46

  Hydrolases (3) · CPC title