Modified L-asparaginase
View Patent ↗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 modified fusion protein comprising
(i) an L-asparaginase comprising the amino acid sequence of SEQ ID NO: 1, and
(ii) one or more polypeptides, wherein the one or more polypeptides comprise the amino acid sequence of SEQ ID NO: 7 or 9.
2. The modified fusion protein according to claim 1 , wherein the L-asparaginase consists of the amino acid sequence of SEQ ID NO: 1.
3. The modified fusion protein according to claim 1 , wherein the modified protein is a fusion protein of the L-asparaginase and the one or more polypeptides.
4. The modified fusion protein according to claim 3 , wherein the one or more polypeptides consist of 200 to 600 proline and alanine amino acid residues.
5. The modified fusion protein according to claim 3 , wherein the one or more polypeptides consist of 200 to 400 proline and alanine amino acid residues, and the proline amino acid residues constitute more than 10% and less than 70% of the polypeptide.
6. The modified fusion protein according to claim 3 , wherein at least one of the one or more polypeptides comprises the amino acid sequence of SEQ ID NO: 7.
7. The modified fusion protein according to claim 3 , wherein at least one of the one or more polypeptides comprises the amino acid sequence of SEQ ID NO: 9.
8. The modified fusion protein according to claim 3 , wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 11.
9. The modified fusion protein according to claim 3 , wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 13.
10. The modified fusion protein according to claim 3 , wherein the L-asparaginase comprises the amino acid sequence of SEQ ID NO: 1 and the polypeptide consists of the amino acid sequence of SEQ ID NO: 7, or 9.
11. The modified fusion protein according to claim 2 , wherein at least one of the one or more polypeptides consists of the amino acid sequence of SEQ ID NO: 7.
12. The modified fusion protein according to claim 2 , wherein at least one of the one or more polypeptides consists of the amino acid sequence of SEQ ID NO: 9.
13. The modified fusion protein according to claim 3 , wherein the L-asparaginase is composed of four subunits.
14. The modified fusion protein according to claim 3 , wherein the fusion protein consists of the amino acid sequence of SEQ ID NO: 11 or 13.
15. A pharmaceutical composition comprising the modified fusion protein according to claim 1 and one or more pharmaceutical acceptable carriers or excipients.
16. The pharmaceutical composition of claim 15 , wherein the one or more polypeptides is present in an amount sufficient to mediate a decreased immunogenicity of the modified protein following administration to a human subject.
17. The pharmaceutical composition of claim 15 , wherein the one or more polypeptides is present in an amount sufficient to increase the plasma half-life of the L-asparaginase following administration to a human subject.