Human methylthioadenosine/adenosine depleting enzyme variants for cancer therapy

Patent metadata

Field	Value
Publication number	US-11396647-B2
Application number	US-202117427519-A
Country	US
Kind code	B2
Filing date	Jan 6, 2021
Priority date	Jan 7, 2020
Publication date	Jul 26, 2022
Grant date	Jul 26, 2022

Abstract

Disclosed herein are compositions related to conjugated polypeptides with MTA/ADO-degrading enzyme activity. The conjugated polypeptides are engineered to allow for maximal conjugation while maintaining catalytic activities. Also disclosed are nucleic acids, expression vectors, and host cells related to the conjugated polypeptides. Further disclosed are methods of using the pharmaceutical formulations comprising above to treat cancer.

First claim

The invention claimed is:

1. A composition comprising a polypeptide having methylthioadenosine phosphorylase activity,
wherein said polypeptide comprises an amino acid sequence with at least 95% sequence identity to at least 100 consecutive amino acids of SEQ ID NO: 1 and comprises amino acids corresponding to Threonine 18, Threonine 197, Serine 178, Valine 233 and Methionine 196 of SEQ ID NO: 1, and
wherein an amino acid corresponding to Lysine 225 or Lysine 238 of SEQ ID NO: 1 is (i) deleted or (ii) substituted with Alanine, Arginine, Asparagine, Aspartic Acid, Cysteine, Glutamine, Glutamic Acid, Glycine, Histidine, Isoleucine, Leucine, Methionine, Phenylalanine, Proline, Serine, Threonine, Tryptophan, Tyrosine, or Valine.

2. The composition of claim 1 , wherein said amino acid corresponding to Lysine 225 is substituted with said Arginine.

3. The composition of claim 1 , wherein said amino acid corresponding to Lysine 238 is substituted with said Arginine.

4. The composition of claim 1 , wherein said polypeptide comprises an amino acid sequence comprising a sequence with at least 95% sequence identity to SEQ ID NO: 3 or SEQ ID NO: 5.

5. The composition of claim 1 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 5.

6. The composition of claim 5 , wherein said polypeptide is conjugated to a polymer.

7. The composition of claim 6 , wherein said polymer is a polyethylene glycol polymer.

8. The composition of claim 7 , wherein said polyethylene glycol polymer has an average molecular weight of about 4,000 kilodaltons (kDa) to about 8,000 kDa.

9. The composition of claim 7 , wherein said composition comprises at least six of said polyethylene glycol polymers.

10. The composition of claim 7 , wherein said composition comprises a plurality of said polypeptides, and wherein a number of said polyethylene glycol polymers per polypeptide comprises a Gaussian distribution with a mode of 8±3 of said polyethylene glycol polymers per polypeptide.

11. The composition of claim 10 , wherein said polypeptide comprises the amino acid sequence of SEQ ID NO: 5.

12. The composition of claim 1 , where said polypeptide is conjugated to a polymer, and wherein said polymer is not conjugated to an amino acid corresponding to either of Lysine 225 or Lysine 238 of SEQ ID NO. 1.

13. The composition of claim 12 , wherein said composition comprises at least six of said polymers.

14. The composition of claim 13 , wherein a k cat /K M of said methylthioadenosine phosphorylase activity of said polypeptide is at least 50% of a k cat /K M of a methylthioadenosine phosphorylase comprising SEQ ID NO: 1.

15. The composition of claim 13 , wherein a k cat /K M of said methylthioadenosine phosphorylase activity of said polypeptide is at least 1.5×10 5 M −1 s −1 .

16. The composition of claim 13 , wherein a Vmax of said methylthioadenosine phosphorylase activity of said polypeptide is at least 50% of a Vmax of said methylthioadenosine phosphorylase activity of a methylthioadenosine phosphorylase comprising SEQ ID NO: 1.

17. The composition of claim 13 , wherein a Km of said methylthioadenosine phosphorylase activity of said polypeptide is no more than twice a Km of said methylthioadenosine phosphorylase activity of a methylthioadenosine phosphorylase comprising SEQ ID NO: 1.

18. The composition of claim 12 , wherein said polypeptide has a serum half-life of at least 36 hours.

19. A composition comprising a population of said polypeptides according to claim 1 , wherein said population comprises trimers of said polypeptides.

Assignees

• Univ Texas

Inventors

• Stone Everett

Classifications

• C12Y204/02028

  S-Methyl-5'-thioadenosine phosphorylase (2.4.2.28) · CPC title

• C12N15/70

  Vectors or expression systems specially adapted for E. coli · CPC title

• C12N11/089

  obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds · CPC title

• C12N9/1077 Primary

  Pentosyltransferases (2.4.2) · CPC title

• C07K2317/76

  Antagonist effect on antigen, e.g. neutralization or inhibition of binding · CPC title