Alpha-Synuclein Kinase

Abstract

The invention provides agents and methods for treatment of diseases associated with Lewy body diseases (LBDs) in the brain of a patient. Preferred agents include inhibitors of PLK2 kinase.

1 . A method for inhibiting phosphorylation of alpha-synuclein in a mammalian cell, the method comprising reducing polo-like kinase 2 (PLK2) activity in the cell such that phosphorylation of synuclein is reduced. 
     
     
         2 . A method for inhibiting phosphorylation of alpha-synuclein in a mammalian cell, the method comprising contacting the cell with a compound that reduces PLK2 activity in the cell such that phosphorylation of alpha-synuclein is reduced. 
     
     
         3 . The method of  claim 2  wherein the agent reduces expression of the PLK2 protein. 
     
     
         4 . The method of  claim 2  wherein the cell is a neuronal cell. 
     
     
         5 . The method of  claim 4  wherein the agent has a molecular weight less than 4000. 
     
     
         6 . The method of  claim 5  wherein the agent is a synthetic compound. 
     
     
         7 . The method of  claim 4  wherein the agent preferentially reduces PLK2 activity activity relative to reduction of PLK1 activity, PLK2 activity, or PLK3 activity. 
     
     
         8 . The method of  claim 7  wherein the agent is a polynucleotide that inhibits expression or translation of a PLK2 RNA transcript. 
     
     
         9 . The method of  claim 8  wherein the agent is an siRNA comprising a double stranded region. 
     
     
         10 . The method of  claim 9  wherein one strand of the double stranded region is perfectly complementary to a PLK2 transcript and lacks perfect complementarity to a PLK1 transcript or a PLK3 transcript. 
     
     
         11 . The method of  claim 5  wherein the compound is 4-[[(7r)-8-cyclopentyl-7-ethyl-5,6,7,8-tetrahydro-5-methyl-6-oxo-2-pteridinyl]amino]-3-methoxy-n-(1-methyl-4-piperidinyl)-benzamide. 
     
     
         12 . A method of treating a patient diagnosed with Parkinson's Disease comprising administering a therapeutically effective amount of an agent that inhibits PLK2 activity. 
     
     
         13 . The method of  claim 12  wherein the agent preferentially inhibits PLK2 activity relative to inhibition of PLK1 activity or PLK3 activity. 
     
     
         14 . The method of  claim 13  wherein the agent preferentially inhibits PLK2 activity relative to inhibition of PLK1 activity and PLK3 activity. 
     
     
         15 . The method of  claim 14  wherein the agent is an siRNA. 
     
     
         16 . The method of  claim 15  wherein the agent is an siRNA comprising a double stranded region. 
     
     
         17 . The method of  claim 16  wherein one strand of the double stranded region is perfectly complementary to a PLK2 transcript and lacks perfect complementarity to a PLK1 transcript or a PLK3 transcript. 
     
     
         18 . The method of  claim 12  wherein the agent is a synthetic compound with a molecular weight less than 4000. 
     
     
         19 . The method of  claim 12  wherein the compound is 4-[[(7r)-8-cyclopentyl-7-ethyl-5,6,7,8-tetrahydro-5-methyl-6-oxo-2-pteridinyl]amino]-3-methoxy-n-(1-methyl-4-piperidinyl)-benzamide. 
     
     
         20 . The method of  claim 12  wherein the patient is not diagnosed or under treatment for cancer. 
     
     
         21 . The method of  claim 12  wherein the patient is not diagnosed or under treatment for Alzheimer's disease. 
     
     
         22 . A method of identifying an agent reduces alpha-synuclein phosphorylation in a mammalian cell expressing alpha-synuclein comprising selecting an agent that
 a) reduces activity of PLK2 in a cell expressing PLK2; and   b) does not reduce activity of PLK1 in a cell expressing PLK1, or reduces activity of PLK1 at a higher EC 50  than for PLK2; and/or   c) does not reduce activity of PLK3 in a cell expressing PLK3, or reduces activity of PLK3 at a higher EC 50  than for PLK2; and/or   d) does not reduce activity of PLK4 in a cell expressing PLK4, or reduces activity of PLK4 at a higher EC 50  than for PLK2.   
     
     
         23 . The method of  claim 22  wherein the cell is a mammalian cell overexpressing alpha-synuclein. 
     
     
         24 . The method  claim 23  comprising selecting an agent that
 a) reduces activity of PLK2 in a cell expressing PLK2;   b) does not reduce activity of PLK1 in a cell expressing PLK1, or reduces activity of PLK1 at a higher EC 50  than for PLK2;   c) does not reduce activity of PLK3 in a cell expressing PLK3, or reduces activity of PLK3 at a higher EC 50  than for PLK2; and   d) does not reduce activity of PLK4 in a cell expressing PLK4, or reduces activity of PLK4 at a higher EC 50  than for PLK2.   
     
     
         25 . A method of identifying an agent for treatment of Parkinson's disease comprising selecting an agent according to the method of  claim 22 , and further comprising determining whether the selected agent shows activity useful in treating Lewy Body Disease in an animal model of the disease. 
     
     
         26 . A method of identifying an agent for treatment of Parkinson's disease comprising selecting an agent according to the method of  claim 22 , and further comprising determining whether the selected agent shows activity useful in treating Lewy Body Disease in a cellular model of the disease. 
     
     
         27 . The method of  claim 26  wherein the cellular model of the disease is a neuronally-derived cell culture. 
     
     
         28 . The method of  claim 27  wherein the cellular model of the disease is a mammalian cell over-expressing alpha-synuclein. 
     
     
         29 . The method of  claim 27  wherein the activity is reduction of the proportion of total alpha-synuclein that is phosphorylated at of serine-129. 
     
     
         30 . The method of  claim 27  wherein said activity is a reduction in aggregation of alpha-synuclein in the cell. 
     
     
         31 . The method of  claim 22 , further comprising identifying an agent that modulates PLK2 in the presence of synphilin.