Methods for SHP1 mediated neuroprotection

• US 20030232749A1
• Filed: 03/11/2003
• Published: 12/18/2003
• Est. Priority Date: 03/11/2002
• Status: Active Grant

• Current Assignee: Ortho-McNeil Pharmaceutical Incorporated (Johnson & Johnson)
• Original Assignee: Ortho-McNeil Pharmaceutical Incorporated (Johnson & Johnson)
• Inventors: Renzi, Michael, Thirumalai, Navneeth, Jolliffe, Linda, Farrell, Francis X.
• Granted Patent: US 7,129,267 B2
• Time in Patent Office: Days
• US Class Current: 514/438
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/00 Medicinal preparations containing organic active ingredients
• CPC Class Codes: A61K 31/381 having five-membered rings
• CPC Class Codes: A61K 31/426 1,3-Thiazoles
• CPC Class Codes: A61K 31/427 not condensed and containing further heterocyclic rings
• CPC Class Codes: A61K 38/1816 Erythropoietin [EPO]
• CPC Class Codes: C12Q 1/44 involving esterase
• CPC Class Codes: G01N 2500/02 Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor)
• CPC Class Codes: G01N 2500/04 Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)
• CPC Class Codes: G01N 2500/10 involving cells
• CPC Class Codes: G01N 33/573 for enzymes or isoenzymes

Abstract

The effect of EPO on the phosphorylation of the EPO receptor, the activation of the MAP Kinase pathway, and the expression of SHP-1 were analyzed. EPO was observed to cause a decrease in the expression of its negative regulator SHP-1. The decrease observed at both the mRNA and protein level was dose dependent and persisted as long as 24 hr following EPO treatment. EPO can down regulate the expression of its own negative regulator as a means for increased potency in neurons. Assays were generated to identify compounds that are useful in regulating SHP1 activity in neural cells.

Claims

1. A method for treating a nervous system condition related to EPOR in a subject in need thereof, comprising the step of administering to the subject a therapeutically effective dose of a composition that decreases the tyrosine phosphatase activity of a SHP1 in a cell of the nerve system of the subject.

2. The method of claim 1 wherein said subject is in need of neuroprotection.

3. The method of claim 1 wherein said condition is the result of a seizure disorder, multiple sclerosis, stroke, hypotension, ischemia, myocardial infarction, inflammation, aging or cognitive dysfunction, radiation damage, cerebral palsy, neurodegenerative disease, Alzheimer'"'"'s disease, Parkinson'"'"'s disease, Leigh disease, AIDS dementia, memory loss, amyotrophic lateral sclerosis, alcoholism, mood disorder, anxiety disorder, attention deficit disorder, autism, Creutzfeld-Jakob disease, brain or -spinal cord trauma, heart-lung bypass, glaucoma, retinal ischernia, retinal trauma.

4. The method of claim 1 wherein said condition is an acute nervous system disease selected from the group consisting of ischemic stroke, hemorrhagic stroke, spinal cord injury and traumatic brain injury.

5. The method of claim 1 wherein said condition is a chronic nervous system disease selected from the group consisting of Alzheimer'"'"'s disease, Parkinson'"'"'s disease, peripheral neuropathies, and cognitive impairment associated with coronary artery bypass graft surgery (CABG) and carotid endarterectomy (CEA).

6. The method of claim 1, wherein said cell of nerve system is a neuronal cell.

7. The method of claim 6, wherein the neuronal cell is selected from the group consisting of a sensory neuron, a motoneuron, and an interneuron.

8. The method of claim 1, wherein said cell of the nervous system is a non-neuronal cell.

9. The method of claim 8, wherein the non-neuronal cell is selected from the group consisting of astrocytes and microglia cells.

10. The method of claim 1, wherein said composition is selected from the group consisting of a compound of formula (I);

11. The method of claim 1, wherein said composition is administered in combination with one or more neuroprotective agents.

12. The method of claim 11, wherein said neuroprotective agent is EPO.

13. A method for treating a nervous system condition related to EPOR in a subject in need thereof, comprising administering to the subject a therapeutically effective dose of a composition that decreases the expression of a SHP1 in a cell of the nervous system of the subject.

14. The method of claim 13 wherein the composition comprises an antisense nucleic acid or a siRNA molecule specific for an SHP1 gene and wherein the antisense nucleic acid or siRNA molecule specifically suppresses SHP1 gene expression in a cell of the nerve system of the subject.

15. A method of identifying a compound useful for treating a nervous system condition related to EPOR, comprising the steps of:

1) contacting a test compound with a SHP1 protein or an active fragment thereof, and

2) determining the ability of the test compound to decrease the tyrosine phosphatase activity of SHP1.

16. The method of claim 15 further comprising the steps of

a) contacting a neuronal cell with the test compound;

b) inducing neurotoxicity in the neuronal cell;

c) assaying the cell survival rate in the presence of the test compound, and comparing the cell survival rate with that of a control wherein the neuronal cell is not treated with the test compound.

17. The method of claim 15 wherein said SHP1 protein or active fragment thereof is substantially purified.

18. The method of claim 15 wherein SHP1 protein or active fragment thereof is expressed from a host cell.

19. A method of identifying a compound useful for treating a nervous system condition related to EPOR, comprising the steps of:

c) contacting a test compound with a regulatory sequence for a SHP1 gene or a cellular component that binds to the regulatory sequence for a SHP1 gene; and

d) determining whether the test compound decreases the expression of a gene controlled by said regulatory sequence.

20. The method of claim 19 further comprising the steps of

a) contacting a neuronal cell with the test compound;

b) inducing neurotoxicity in the neuronal cell;

c) assaying the cell survival rate in the presence of the test compound, and comparing the cell survival rate with that of a control wherein the neuronal cell is not treated with the test compound.

21. The method of claim 19 wherein the gene controlled by the SHP1 regulatory sequence is a reporter gene.

22. The method of claim 19 wherein the gene controlled by the SHP1 regulatory sequence is an SHP1 gene.

23. A method of identifying a compound useful for treating a nervous system condition related to EPOR, comprising the steps of:

e) combining a test compound, a labeled ligand for a SHP1 protein, and a SHP1 protein or an active fragment thereof; and

f) measuring the binding of the test compound to the SHP1 protein or active fragment thereof by a reduction in the amount of labeled ligand binding to the SHP1 protein or active fragment thereof.

24. The method of claim 23 further comprising the steps of

a) contacting a neuronal cell with the test compound;

b) inducing neurotoxicity in the neuronal cell;

c) assaying the cell survival rate in the presence of the test compound, and comparing the cell survival rate with that of a control wherein the neuronal cell is not treated with the test compound.

25. A method for treating a nervous system condition related to EPOR in a subject in need thereof comprising administering to the subject a composition comprising one or more inhibitors of a SHP1 tyrosine phosphates protein.