# US 2019/0169589 A1 · Modified L-Asparaginase > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2019/0169589 A1 - Document ID: US20190169589A1 - Status: Published application - Kind: A1 - Application number: 16/226,499 - Filed: 2018-12-19 - Published: 2019-06-06 - Earliest claimed: 2017-06-21 - Assignees: Xl-Protein GmbH; Jazz Pharmaceutical Ireland Limited - Inventors: Lars Friedrich; Anne O'Donnell - Web page: https://patentgrep.com/patent/US20190169589A1 ## CPC - C12N 9/82: Asparaginase {(3.5.1.1)} - C12Y 305/01001: Asparaginase (3.5.1.1) - A61K 38/02: Peptides of undefined number of amino acids; Derivatives thereof - A61K 38/48: acting on peptide bonds (3.4) - C07K 14/00: Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof - C07K 2319/00: Fusion polypeptide - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C12Y: ENZYMES - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract 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. ## Claims ### Claim 1 A modified protein comprising (i) an L-asparaginase and (ii) one or more polypeptides, wherein the one or more polypeptides consists solely of proline and alanine amino acid residues. ### Claim 2 (depends on 1) The modified protein according to claim 1, wherein the L-asparaginase has at least 85% sequence identity to the amino acid sequence of SEQ ID NO: 1. ### Claim 3 (depends on 1) The modified protein according to claim 1, wherein the L-asparaginase has at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 1. ### Claim 4 (depends on 1) The modified protein according to claim 1, wherein the L-asparaginase has at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 1. ### Claim 5 (depends on 1) The modified protein according to claim 1, wherein the L-asparaginase comprises the amino acid sequence of SEQ ID NO: 1. ### Claim 6 (depends on 1) The modified protein according to claim 1, wherein the L-asparaginase consists of the amino acid sequence of SEQ ID NO: 1. ### Claim 7 (depends on 1) The modified protein according to claim 1, wherein the modified protein is a fusion protein of the L-asparaginase and the polypeptide. ### Claim 8 (depends on 7) The modified protein according to claim 7, wherein polypeptide consists of 100 to 600 proline and alanine amino acid residues. ### Claim 9 (depends on 7) The modified protein according to claim 7, wherein the polypeptide consists 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. ### Claim 10 (depends on 7) The modified protein according to claim 7, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 5. ### Claim 11 (depends on 7) The modified protein according to claim 7, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 7. ### Claim 12 (depends on 7) The modified protein according to claim 7, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 9. ### Claim 13 (depends on 7) The modified protein according to claim 7, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 11. ### Claim 14 (depends on 7) The modified protein according to claim 7, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 13. ### Claim 15 (depends on 7) The modified protein according to claim 7, wherein the L-asparaginase comprises the amino acid sequence of SEQ ID NO: 1 and the polypeptide comprises the amino acid sequence of SEQ ID NO: 5, 7, 9, 11 or 13. ### Claim 16 (depends on 1) The modified protein according to claim 1, wherein each of the one or more polypeptides is independently a peptide RN-(P/A)-Rc, wherein (P/A) is an amino acid sequence consisting solely of proline and alanine amino acid residues, RN is a protecting group attached to the N-terminal amino group of the amino acid sequence, and RC is an amino acid residue bound via its amino group to the C-terminal carboxy group of the amino acid sequence, the each of the one or more polypeptides is conjugated to the L-asparaginase via an amide linkage formed from the carboxy group of the C-terminal amino acid residue Rc of the peptide and a free amino group of the L-asparaginase, and at least one of the free amino groups, which the one or more polypeptide(s) are conjugated to, is not an N-terminal α-amino group of the L-asparaginase. ### Claim 17 (depends on 16) The modified protein according to claim 16, wherein the each of the one or more polypeptides independently consists of 15 to 45 proline and alanine amino acid residues. ### Claim 18 (depends on 16) The modified protein according to claim 16, wherein the each of the one or more polypeptides independently consists of 20 to 40 proline and alanine amino acid residues, and the proline amino acid residues constitute more than 10% and less than 70% of the amino acid sequence. ### Claim 19 (depends on 16) The modified protein according to claim 16, wherein at least one of the one or more polypeptides comprises the amino acid sequence of SEQ ID NO: 5. ### Claim 20 (depends on 16) The modified protein according to claim 16, wherein at least one of the one or more polypeptides comprises the amino acid sequence of SEQ ID NO: 15. ### Claim 21 (depends on 16) The modified protein according to claim 16, wherein RN is pyroglutamoyl or acetyl, and/or RC is ε-aminohexanoic acid. ### Claim 22 (depends on 16) The modified protein according to claim 16, wherein (i) at least one of the free amino groups, which the one or more polypeptides are conjugated to, is an ε-amino group of a lysine residue of the L-asparaginase; or (ii) the free amino groups, which the one or more polypeptides are conjugated to, are selected from the group consisting of the ε-amino group(s) of any lysine residue(s) of the L-asparaginase and the N-terminal α-amino group(s) of the L-asparaginase. ### Claim 23 (depends on 16) The modified protein according to claim 16, wherein the L-asparaginase is composed of four subunits, and 9 to 13 polypeptides are conjugated to each subunit of the L-asparaginase. ### Claim 24 (depends on 16) The modified protein according to claim 16, wherein the L-asparaginase comprises the amino acid sequence of SEQ ID NO: 1 and the one or more polypeptides comprise the amino acid sequence of SEQ ID NO: 5 or 15. ### Claim 25 (depends on 1) A pharmaceutical composition comprising the modified protein according to claim 1 and one or more pharmaceutical acceptable carriers or excipients. ### Claim 26 (depends on 25) The pharmaceutical composition of claim 25, 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. ### Claim 27 (depends on 25) The pharmaceutical composition of claim 25, 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.