Compositions and methods of using chondroitinase ABCI mutants
View Patent ↗The present invention relates to protein and nucleic acid mutants of chondroitinase ABCI. Such chondroitinase ABCI mutant enzymes exhibit altered chondroitin lyase activity or increased resistance to inactivation from stressors including UV light or heat. Methods of using chondroitinase ABCI mutant enzymes are also provided.
1. An isolated mutant chondroitinase ABCI enzyme selected from SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO: 6.
2. The mutant chondroitinase ABCI enzyme of claim 1 , wherein said enzyme comprises SEQ ID NO: 1.
3. The mutant chondroitinase ABCI enzyme of claim 1 , wherein said enzyme comprises SEQ ID NO:2.
4. A method of treating a central nervous system injury comprising administering a therapeutically effective amount of an enzyme selected from SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO: 6.
5. The method of claim 4 , wherein the mutant chondroitinase ABCI enzyme is administered following a contusion injury to the central nervous system.
6. The method of claim 4 , wherein the mutant chondroitinase ABCI enzyme is administered following a non-contusion injury to the central nervous system.
7. The method of claim 4 , wherein the mutant chondroitinase ABCI enzyme is administered following a spinal cord injury.
8. The method of claim 4 , wherein the mutant chondroitinase ABCI enzyme is administered locally.
9. The method of claim 8 , wherein the local administration is selected from intrathecal and topical administration.
10. The method of claim 4 , wherein said enzyme comprises SEQ ID NO:1.
11. The method of claim 4 , wherein said enzyme comprises SEQ ID NO:2.
12. A method of promoting neuronal outgrowth comprising administering a therapeutically effective amount of an enzyme selected from SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO: 6.
13. The method of claim 12 , wherein the mutant chondroitinase ABCI enzyme is administered following a contusion injury to the central nervous system.
14. The method of claim 12 , wherein the mutant chondroitinase ABCI enzyme is administered following a non-contusion injury to the central nervous system.
15. The method of claim 12 , wherein the mutant chondroitinase ABCI enzyme is administered following a spinal cord injury.
16. The method of claim 12 , wherein the mutant chondroitinase ABCI enzyme is administered locally.
17. The method of claim 16 , wherein the local administration is selected from intrathecal and topical administration.
18. The method of claim 12 , wherein said enzyme comprises SEQ ID NO:1.
19. The method of claim 12 , wherein said enzyme comprises SEQ ID NO:2.
20. A method of facilitating diffusion of an agent in a central nervous system comprising:
administering the agent to the central nervous system; and
administering to the central nervous system at least one enzyme selected from SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO: 6.
21. The method of claim 20 , wherein the at least one enzyme comprises SEQ ID NO:1.
22. The method of claim 20 , wherein the at least one enzyme comprises SEQ ID NO:2.
23. The method of claim 20 , wherein the at least one enzyme is administered in an effective amount.
24. The method of claim 20 , wherein the at least one enzyme is administered with one or more pharmaceutical carriers.
25. The method of claim 20 , wherein the pharmaceutical carrier is selected from the group consisting of diluents, binders, lubricants, coloring agents, disintegrating agents, buffer agents, isotonizing agents, polyols, preservants, and anesthetics.
26. The method of claim 20 , wherein the agent is selected from the group consisting of complex biologics, small molecules, and transplanted cells.
27. A method of enhancing uptake of an agent in a central nervous system, comprising administering the agent and at least one enzyme capable of cleaving chondroitin sulfate proteoglycans, wherein the at least one enzyme is selected from SEQ ID NO: 1, SEQ ID NO:2, and SEQ ID NO: 6.
28. The method of claim 27 , wherein the at least one enzyme comprises SEQ ID NO:1.
29. The method of claim 27 , wherein the at least one enzyme comprises SEQ ID NO:2.
30. The method of claim 27 , wherein the at least one enzyme is administered in an effective amount.
31. The method of claim 27 , wherein the at least one enzyme is administered with one or more pharmaceutical carriers.
32. The method of claim 27 , wherein the pharmaceutical carrier is selected from the group consisting of diluents, binders, lubricants, coloring agents, disintegrating agents, buffer agents, isotonizing agents, polyols, preservants, and anesthetics.
33. The method of claim 27 , wherein the agent is selected from the group consisting of complex biologics, small molecules, and transplanted cells.
34. A method for inhibiting extravasation of cells from blood vessels comprising administering to a patient an enzyme that cleaves a chondroitin sulfate proteoglycan, wherein the enzyme is selected from SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO: 6.
35. The method of claim 34 , wherein the cells are selected from the group consisting of white blood cells, leukocytes, neutrophils, eosinophils, basophils, lymphocytes, B-cells, T-cells, monocytes, and macrophages.
36. A method of treating inflammation in a patient comprising administering to the patient an enzyme that cleaves a chondroitin sulfate proteoglycans, wherein the enzyme is selected from SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO: 6.
37. A method of treating inflammation in a patient comprising: extracting cells circulating in the blood stream of the patient; subjecting the cells to an enzyme that cleaves chondroitin sulfate proteoglycan ex vivo to modify the cells; and administering the modified cells to the patient, wherein the enzyme is selected from SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO: 6.
38. The mutant chondroitinase ABCI enzyme of claim 1 , wherein the enzyme comprises SEQ ID NO: 6.
39. The method of claim 4 , wherein the enzyme comprises SEQ ID NO: 6.
40. The method of claim 12 , wherein the enzyme comprises SEQ ID NO: 6.
41. The method of claim 20 , wherein the at least one enzyme comprises SEQ ID NO: 6.
42. The method of claim 27 , wherein the at least one enzyme comprises SEQ ID NO: 6.
43. The method of claim 34 , wherein the enzyme comprises SEQ ID NO: 1.
44. The method of claim 34 , wherein the enzyme comprises SEQ ID NO: 2.
45. The method of claim 34 , wherein the enzyme comprises SEQ ID NO: 6.
46. The method of claim 36 , wherein the enzyme comprises SEQ ID NO: 1.
47. The method of claim 36 , wherein the enzyme comprises SEQ ID NO: 2.
48. The method of claim 36 , wherein the enzyme comprises SEQ ID NO: 6.
49. The method of claim 37 , wherein the enzyme comprises SEQ ID NO: 1.
50. The method of claim 37 , wherein the enzyme comprises SEQ ID NO: 2.
51. The method of claim 37 , wherein the enzyme comprises SEQ ID NO: 6.