USE OF INHIBITORS OF PHOSPHOLIPASE A2 FOR THE TREATMENT, PREVENTION OR DIAGNOSIS OF NEURAL INFLAMMATORY OR DEMYELINATING DISEASE
(19)AUSTRALIAN PATENT OFFICE(54) Title USE OF INHIBITORS OF PHOSPHOLIPASE A2 FOR THE TREATMENT, PREVENTION ORDIAGNOSIS OF NEURAL INFLAMMATORY OR DEMYELINATING DISEASE(51)6 International Patent Classification(s) A61K 045/00 G01N The present invention provides methods of preventing and treating neural inflammatory or demyelinating disease, such as multiple sclerosis, via an inhibition of the activity or expression of phospholipase A2. The invention further relates to methods of identifying phospholipase A2 inhibitors and their use thereof for the prevention and/or treatment of neural inflammatory or demyelinating disease. An observed increase in the amount of phospholipase A2 in neural lesions in the EAE animal model system indicates that elevated phospholipase A2 activity or levels correlate with neural inflammatory or demyelinating disease. Therefore, in a further aspect the invention provides methods for the diagnosis and prognostication of neural inflammatory or demyelinating disease, such as multiple sclerosis. CLAIMS : 1. A method of preventing or treating a neural inflammatory or demyelinating disease in an animal, said method comprising inhibiting the activity of a phospholipase A2 in the animal.
2. The method of claim 1, wherein the animal is a mammal.
3. The method of claim 1, wherein the animal is a human.
4. The method of claim 1, wherein the neural inflammatory or demyelinating disease is selected from the group consisting of Multiple Sclerosis, Alzheimer's disease, amyotrophic lateral sclerosis and stroke.
5. The method of claim 1, wherein the phospholipase A2 is a cytosolic phospholipase A2.
6. The method of claim 1, wherein the method comprises administering to the animal an effective amount of a phospholipase A2 inhibitor.
7. The method of claim 1, wherein the inhibitor is selected from the group consisting of arachidonic acid analogues, benzenesulfonamide derivatives, bromoenol lactone, p- bromophenyl bromide, bromophenacyl bromide, trifluoromethylketones, sialoglycolipids and proteoglycans.
8. The method of claim 7, wherein the inhibitor is selected from the group consisting of arachidonyl trifluoromethyl ketone, methyl arachidonyl fluorophosphonate, palmitoyl trifluoromethyl ketone. <Desc/Clms Page number 70> 9. The method of claim 1, wherein the method comprises inhibiting the expression of a phospholipase A2.
10. The method of claim 9, wherein the method comprises administering to the animal an effective amount of a phospholipase A2 inhibitor.
11. The method of claim 10 wherein said phospholipase A2 inhibitor is selected from the group consisting of an antisense molecule and an siRNA or siRNA-like molecule.
12. The method of claim 11 wherein the antisense molecule is a nucleic acid that is substantially complementary to a portion of an mRNA encoding a phospholipase A2.
13. The method of claim 12 wherein the antisense molecule is complementary to a portion of a nucleic acid sequence substantially identical to a sequence selected from the group consisting of SEQ ID NO : 1 and SEQ ID NO : 3.
14. The method of claim 12 wherein the portion of an mRNA comprises at least 5 contiguous bases.
15. The method of claim 10 wherein the siRNA or siRNA-like molecule is substantially identical to a portion of an mRNA encoding a phospholipase A2.
16. The method of claim 10 wherein the siRNA or siRNA-like molecule is substantially identical to a portion of an mRNA corresponding to a DNA sequence selected from the group consisting of SEQ ID NO : 1 and SEQ ID NO : 3. <Desc/Clms Page number 71> 17. The method of claim 10 wherein the siRNA or siRNA-like molecule comprises less than about 30 nucleotides.
18. The method of claim 17 wherein the siRNA or siRNA-like molecule comprises about 21 to about 23 nucleotides.
19. Use of a phospholipase A2 inhibitor for preventing or treating a neural inflammatory or demyelinating disease in an animal..
20. Use of a phospholipase A2 inhibitor for preparation of a medicament for preventing or treating a neural inflammatory or demyelinating disease in an animal.
21. The use of claim 19, wherein the animal is a mammal.
22. The use of claim 21, wherein the mammal is a human.
23. The use of claim 19, wherein the neural inflammatory or demyelinating disease is selected from the group consisting of Multiple Sclerosis, Alzheimer's disease, amyotrophic lateral sclerosis and stroke.
24. The use of claim 19, wherein the phospholipase A2 is a cytosolic phospholipase A2.
25. The use of claim 19, wherein the inhibitor is selected from the group consisting of arachidonic acid analogues, benzenesulfonamide derivatives, bromoenol lactone, p- bromophenyl bromide, bromophenacyl bromide, trifluoromethylketones, sialoglycolipids and proteoglycans. <Desc/Clms Page number 72> 26. The use of claim 21, wherein the inhibitor is selected from the group consisting of arachidonyl trifluoromethyl ketone, methyl arachidonyl fluorophosphonate, palmitoyl trifluoromethyl ketone.
27. The use of claim 19, wherein the inhibitor is an inhibitor of phospholipase A2 expression.
28. The use of claim 27 wherein said phospholipase A2 inhibitor is selected from the group consisting of an antisense molecule and an siRNA or siRNA-like molecule.
29. The use of claim 28 wherein the antisense molecule is. a nucleic acid that is substantially complementary to a portion of an mRNA encoding a phospholipase A2.
30. The use of claim 28 wherein the antisense molecule is complementary to a portion of a nucleic acid sequence substantially identical to a sequence selected from the group consisting of SEQ ID NO : 1 and SEQ ID NO : 3.
31. The use of claim 29 wherein the portion of an mRNA comprises at least 5 contiguous bases.
32. The use of claim 28 wherein the siRNA or siRNA-like molecule is substantially identical to a portion of an mRNA encoding a phospholipase A2.
33. The use of claim 28 wherein the siRNA or siRNA-like molecule is substantially identical to a portion of an mRNA corresponding to a DNA sequence selected from the group consisting of SEQ ID NO : 1 and SEQ ID NO : 3. <Desc/Clms Page number 73> 34. The use of claim 28 wherein the siRNA or siRNA-like molecule comprises less than about 30 nucleotides.
35. The use of claim 34 wherein the siRNA or siRNA-like molecule comprises about 21 to about 23 nucleotides.
36. A commercial package comprising a phospholipase A2 inhibitor together with instructions for preventing or treating a neural inflammatory or demyelinating disease in an animal.
37. The commercial package of claim 36, wherein the animal is a mammal.
38. The commercial package of claim 37, wherein the mammal is a human.
39. The commercial package of claim 36, wherein the neural inflammatory or demyelinating disease is selected from the group consisting of Multiple Sclerosis, Alzheimer's disease, amyotrophic lateral sclerosis and stroke.
40. The commercial package of claim 36, wherein the phospholipase A2 is a cytosolic phospholipase A2.
41. The commercial package of claim 36, wherein the inhibitor is selected from the group consisting of arachidonic acid analogues, benzenesulfonamide derivatives, bromoenol lactone, p-bromophenyl bromide, bromophenacyl bromide, trifluoromethylketones, sialoglycolipids and proteoglycans.
42. The commercial package of claim 36, wherein the inhibitor is selected from the group consisting of arachidonyl <Desc/Clms Page number 74> trifluoromethyl ketone, methyl arachidonyl fluorophosphonate, palmitoyl trifluoromethyl ketone.
43. The commercial package of claim 36, wherein the inhibitor is an inhibitor of phospholipase A2 expression.
44. The commercial package of claim 43 wherein said phospholipase A2 inhibitor is selected from the group consisting of an antisense molecule and an siRNA or siRNA-like molecule.
45. The commercial package of claim 44 wherein the antisense molecule is a nucleic acid that is substantially complementary to a portion of an mRNA encoding a phospholipase A2- 46. The commercial package of claim 44 wherein the antisense molecule is complementary to a portion of a nucleic acid sequence substantially identical to a sequence selected from the group consisting of SEQ ID NO : 1 and SEQ ID NO : 3.
47. The commercial package of claim 45 wherein the portion of an mRNA comprises at least 5 contiguous bases.
48. The commercial package of claim 44 wherein the siRNA or siRNA-like molecule is substantially identical to a portion of an mRNA encoding a phospholipase A2.
49. The commercial, package of claim 44 wherein the siRNA or siRNA-like molecule is substantially identical to a portion of an mRNA corresponding to a DNA sequence selected from the group consisting of SEQ ID NO : 1 and SEQID NO : 3. <Desc/Clms Page number 75> 50. The commercial package of claim 44 wherein the siRNA or siRNA-like molecule comprises less than about 30 nucleotides.
51. The commercial package of claim 50 wherein the siRNA or siRNA-like molecule comprises about 21 to about 23 nucleotides.
52. A method of identifying a compound for prevention and/or treatment of neural inflammatory or demyelinating disease, said method comprising: (a) providing a test compound ; and (b) determining whether activity or expression of a phospholipase A2 is decreased in the presence of said test compound ; wherein a decrease in said activity is indicative that said test compound may be used for treating a neural inflammatory or demyelinating disease.
53. The method of claim 52, further comprising the step of assaying the compounds for activity in the prevention or treatment of a neural inflammatory or demyelinating disease.
54. The method of method of claim 52, wherein the phospholipase A2 is a mammalian phospholipase As.
55. The method of claim 54, wherein the phospholipase A2 is a human phospholipase A2- 56. The method of claim 52, wherein the phospholipase A2 is a cytosolic phospholipase A2. <Desc/Clms Page number 76> 57. The method of claim 52, wherein the neural inflammatory or demyelinating disease is selected from the group consisting of Multiple Sclerosis, Alzheimer's disease, amyotrophic lateral sclerosis and stroke.
58. A method of assessing a neural inflammatory or demyelinating disease in an animal, said method comprising : (a) determining a test level of phospholipase A2 protein or phospholipase A2 encoding mRNA or phospholipase A2 enzyme activity in tissue or body fluid of the animal; and (b) comparing said test level of phospholipase A2 protein or phospholipase A2 encoding mRNA or phospholipase A2 activity to an established standard; or to a corresponding level of phospholipase A2 protein or phospholipase A2 encoding mRNA or phospholipase A2 enzyme activity in tissue or body fluid of a control animal; or to a corresponding level of phospholipase As protein or phospholipase A2 encoding mRNA or phospholipase A2 enzymatic activity in tissue or body fluid obtained from said animal at an earlier time;STDC0109 wherein an increase in said test level is indicative of the neural inflammatory or demyelinating disease.
59. The method of claim 58, wherein the animal is a mammal.
60. The method of claim 58, wherein the animal is a human.
61. The method of claim 58, wherein the phospholipase As is a cytosolic phospholipase A2.
62. The method of claim 58, wherein the neural inflammatory or demyelinating disease is selected from the group <Desc/Clms Page number 77> consisting of Multiple Sclerosis, Alzheimer's disease, amyotrophic lateral sclerosis and stroke.
63. The method of claim 58, wherein the tissue or body fluid is selected from the group consisting of blood, plasma, cerebrospinal fluid, neural cells, endothelia, and immune cells.
64. The commercial package of claim 63, wherein said immune cells are selected from the group consisting of macrophages, leukocytes and lymphocytes.
65. A commercial package comprising means for determining the level of phospholipase A2 protein or phospholipase As encoding mRNA or phospholipase As enzyme activity in a tissue or body fluid of an animal together with instructions for assessing a neural inflammatory or demyelinating disease.
66. The commercial package of claim 65, wherein said instructions comprise: (a) determining, using said means, a test level of phospholipase A2 protein or phospholipase A2 encoding mRNA or phospholipase A2 enzyme activity in tissue or body fluid of the animal; and (b) comparing said test level of phospholipase A2 protein or phospholipase As encoding mRNA or phospholipase A2 activity to an established standard; or to a corresponding level of phospholipase A2 protein or phospholipase As encoding mRNA or phospholipase A2 enzyme activity in tissue or body fluid of a control animal;STDC0386 or to a corresponding level of phospholipase A2 protein or phospholipase A2 encoding mRNA or phospholipase As enzymatic activity in tissue or body fluid obtained from <Desc/Clms Page number 78> said animal at an earlier time ; wherein an increase in said test level is indicative of the neural inflammatory or demyelinating disease.
67. The commercial package of claim 65, wherein the animal is a mammal.
68. The commercial package of claim 65, wherein the animal is a human.
69. The commercial package of claim 65, wherein the phospholipase A2 is a cytosolic phospholipase A2.
70. The commercial package of claim 65, wherein the neural . inflammatory or demyelinating disease is selected from the group consisting of Multiple Sclerosis, Alzheimer's disease, amyotrophic lateral sclerosis and stroke.
71. The commercial package of claim 65, wherein the tissue or body fluid is selected from the group consisting of blood, plasma, cerebrospinal fluid, neural cells, endothelia, and immune cells.
72. The commercial package of claim 71, wherein said immune cells are selected from the group consisting of macrophages, leukocytes and lymphocytes.
73. A method of identifying or characterizing a compound for prevention or treatment of a neural inflammatory or demyelinating disease, said method comprising : (a) contacting a test compound with a cell comprising a first nucleic acid comprising a transcriptionally regulatory element normally associated'with a PLA2 gene, operably linked to a second nucleic acid <Desc/Clms Page number 79> comprising a reporter gene capable of encoding a reporter protein; and (b) determining whether reporter gene expression or reporter protein activity is decreased in the presence of said test compound; said decrease in reporter gene expression or reporter protein activity being an indication that said test compound may be used for prevention or treatment of neural inflammatory or demyelinating disease.