Claims for Patent: 9,051,561

Summary for Patent: 9,051,561

Title:	Asparaginase and treating diseases associated with asparagine dependence
Abstract:	Disclosed herein are polypeptides which exhibit asparaginase activity and little to no glutaminase activity. The polypeptides have sequences which correspond to W. succinogenes asparaginase, but have an amino acid residue other than proline at amino acid position 121. The polypeptides exhibit higher kinetic rates of asparaginase activity to glutaminase activity as compared to W. succinogenes asparaginase and Elspar.RTM., an E. coli asparaginase.
Inventor(s):	Durden; Donald (La Jolla, CA)
Assignee:	Children\'s Hospital Los Angeles (Los Angeles, CA)
Application Number:	14/348,637
Patent Claims:	1. A recombinantly produced polypeptide comprising an amino acid sequence that corresponds to the protein having SEQ ID NO: 10 and has an amino acid residue other than proline at amino acid position 121, wherein said polypeptide, which has asparaginase activity and exhibits no glutaminase activity or reduced glutaminase activity is expressed in the periplasm of an E. coli host cell. 2. A recombinantly produced polypeptide having an amino acid sequence that has at least 90-100%, 91-100%, 92-100%, 93-100%, 94-100%, 95-100%, 96-100%, 97-100%, 98-100%, 99-100%, or 100% sequence identity to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4, wherein said polypeptide, which has asparaginase activity and exhibits no glutaminase activity or reduced glutaminase activity is expressed in the periplasm of an E. coli host cell. 3. The recombinantly produced polypeptide of claim 1, and further comprising a signal sequence having SEQ ID NO: 5 or SEQ ID NO: 6. 4. The recombinantly produced polypeptide of claim 1, wherein the amino acid residue is serine, alanine, glycine, cysteine, or threonine. 5. The recombinantly produced polypeptide of claim 1, wherein the polypeptide exhibits a kinetic rate of asparaginase activity that is greater than about 7.5 times that of its kinetic rate of glutaminase activity. 6. An isolated nucleic acid molecule which encodes the polypeptide of claim 1. 7. A host cell which contains the isolated nucleic acid molecule of claim 6. 8. A pharmaceutical composition comprising as an active pharmaceutical ingredient, the polypeptide according to claim 1, and a pharmaceutically acceptable carrier. 9. The pharmaceutical composition according to claim 8, wherein the polypeptide is provided in a therapeutically effective amount for treating a disease which responds to asparagine depletion and/or asparaginase therapy. 10. A method of treating a patient having a disease which comprises administering to the patient a therapeutically effective amount of a recombinantly produced polypeptide according to claim 1 to thereby result in an observable improvement in the clinical symptoms of the disease. 11. A kit which comprises at least one polypeptide according to claim 1 packaged together with a drug delivery device. 12. The recombinantly produced polypeptide of claim 1, wherein an SRP signal sequence was used to express the polypeptide. 13. The recombinantly produced polypeptide of claim 1, wherein a MalE signal sequence was used to express the polypeptide. 14. The recombinantly produced polypeptide of claim 1, wherein a melA promoter was used to express the polypeptide. 15. The recombinantly produced polypeptide of claim 1, wherein a rhaB promoter was used to express the polypeptide. 16. The recombinantly produced polypeptide of claim 1, wherein an SRP signal sequence and a melA promoter was used to express the polypeptide. 17. The recombinantly produced polypeptide of claim 1, wherein the polypeptide exhibits a kinetic rate of asparaginase activity that is 7.6 to about 21.8 times that of its kinetic rate of glutaminase activity. 18. The recombinantly produced polypeptide of claim 1, wherein the polypeptide exhibits a kinetic rate of asparaginase activity that is between 7.5 to 22 times that of its kinetic rate of glutaminase activity. 19. The recombinantly produced polypeptide of claim 2, and further comprising a signal sequence having SEQ ID NO: 5 or SEQ ID NO: 6. 20. The recombinantly produced polypeptide of claim 2, wherein the amino acid residue is serine, alanine, glycine, cysteine, or threonine. 21. The recombinantly produced polypeptide of claim 2, wherein the polypeptide exhibits a kinetic rate of asparaginase activity that is greater than about 7.5 times that of its kinetic rate of glutaminase activity. 22. An isolated nucleic acid molecule which encodes the polypeptide of claim 2. 23. A host cell which contains the isolated nucleic acid molecule of claim 22. 24. A pharmaceutical composition comprising as an active pharmaceutical ingredient, the polypeptide according to claim 2, and a pharmaceutically acceptable carrier. 25. The pharmaceutical composition according to claim 24, wherein the polypeptide is provided in a therapeutically effective amount for treating a disease which responds to asparagine depletion and/or asparaginase therapy. 26. A method of treating a patient having a disease which comprises administering to the patient a therapeutically effective amount of an isolated and/or recombinantly produced polypeptide according to claim 2 to thereby result in an observable improvement in the clinical symptoms of the disease. 27. A kit which comprises at least one polypeptide according to claim 2 packaged together with a drug delivery device. 28. The recombinantly produced polypeptide of claim 2, wherein an SRP signal sequence was used to express the polypeptide. 29. The recombinantly produced polypeptide of claim 2, wherein a MalE signal sequence was used to express the polypeptide. 30. The recombinantly produced polypeptide of claim 2, wherein a melA promoter was used to express the polypeptide. 31. The recombinantly produced polypeptide of claim 2, wherein a rhaB promoter was used to express the polypeptide. 32. The recombinantly produced polypeptide of claim 2, wherein an SRP signal sequence and a melA promoter was used to express the polypeptide. 33. The recombinantly produced polypeptide of claim 2, wherein the polypeptide exhibits a kinetic rate of asparaginase activity that is 7.6 to about 21.8 times that of its kinetic rate of glutaminase activity. 34. The recombinantly produced polypeptide of claim 2, wherein the polypeptide exhibits a kinetic rate of asparaginase activity that is between 7.5 to 22 times that of its kinetic rate of glutaminase activity. 35. The recombinantly produced polypeptide of claim 1 wherein the amino acid residue is serine. 36. The recombinantly produced polypeptide of claim 2 wherein the amino acid residue is serine.

Details for Patent 9,051,561

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Recordati Rare Diseases, Inc.	ELSPAR	asparaginase	For Injection	101063	01/10/1978		2039-02-26