Modified L-asparaginase
The disclosure provides a modified protein that is a combination of (i) an L-asparaginase and (ii) one or more (poly)peptide(s), wherein the (poly)peptide consists solely of proline and alanine amino acid residues, and methods of preparation and use thereof.
1. A method of treating cancer in a human subject in need thereof comprising administering to the human subject a recombinant protein, wherein the recombinant protein is a tetramer, and each monomer of the tetramer comprises:
(i) an L-asparaginase subunit comprising SEQ ID NO: 1 fused to
(ii) a polypeptide comprising about 100 to 600 proline and alanine amino acid residues.
2. The method according to claim 1 , wherein the L-asparaginase subunit has an amino acid sequence consisting of SEQ ID NO: 1.
3. The method according to claim 1 , wherein the polypeptide consists of about 100 to 600 proline and alanine amino acid residues.
4. The method according to claim 1 , wherein the proline amino acid residues constitute more than 10% and less than 70% of the polypeptide.
5. The method according to claim 1 , wherein the polypeptide has an amino acid sequence comprising SEQ ID NO: 5.
6. The method according to claim 1 , wherein the polypeptide has an amino acid sequence comprising SEQ ID NO: 7.
7. The method according to claim 1 , wherein the polypeptide has an amino acid sequence comprising SEQ ID NO: 9.
8. The method according to claim 1 , wherein the monomer has an amino acid sequence comprising SEQ ID NO: 11.
9. The method according to claim 1 , wherein the monomer has an amino acid sequence comprising SEQ ID NO: 13.
10. The method according to claim 1 , wherein the recombinant protein is present in a pharmaceutical formulation with one or more pharmaceutically acceptable carriers or excipients.
11. The method of claim 10 , wherein the recombinant protein is present in an amount sufficient to mediate a decreased immunogenicity of the L-asparaginase subunit following administration to the human subject.
12. The method of claim 10 , wherein the recombinant protein is present in an amount sufficient to increase the plasma half-life of the L-asparaginase subunit following administration to a human subject.
13. The method of claim 1 , wherein the cancer is leukemia or non-Hodgkin's lymphoma.
14. The method of claim 1 , wherein the polypeptide is fused at the N-terminus of the L-asparaginase subunit.
15. The method of claim 1 , wherein the cancer is pancreatic cancer.
16. The method according to claim 1 , wherein no more than 6 consecutive amino acid residues in the polypeptide are identical.
17. The method of claim 13 , wherein the cancer is leukemia.
18. The method of claim 17 , wherein the leukemia is acute lymphoblastic leukemia (ALL).
19. The method of claim 17 , wherein the leukemia is acute myeloid leukemia (AML).
20. The method of claim 1 , wherein between about 1 U/kg and 25 U/kg of the recombinant protein are administered to the human subject.
21. The method of claim 1 , wherein the administering is repeated from about twice a week to once a month.
22. The method of claim 1 , wherein the polypeptide is fused to the L-asparaginase subunit at its N-terminus or its C-terminus.
23. The method of claim 1 , wherein the polypeptide of (ii) increases the activity of the L-asparaginase.
24. The method of claim 1 , wherein the polypeptide comprises about 100 to 400 proline and alanine amino acid residues.
25. The method of claim 1 , wherein the polypeptide comprises about 200 to 600 proline and alanine amino acid residues.
26. The method of claim 1 , wherein the polypeptide comprises about 200 to 400 proline and alanine amino acid residues.