Phosphodiesterase activity and regulation of phosphodiesterase 1B-mediated signaling in brain

Abstract

The present invention provides methods and compositions for modulating the activity of phosphodiesterase 1B (PDE1B) in intracellular signaling pathways, including but not limited to, dopamine D1 intracellular signaling pathways. The invention also provides methods and compositions for modulating the activities of intracellular signaling molecules, including, but not limited to, DARPP-32 and GluR1 AMPA receptor, via modulation of PDE1B. The invention also provides pharmaceutical compositions and methods of screening for compounds that modulate PDE1B activity. The invention also provides methods of treating or ameliorating the symptoms of a disorder, including but not limited to a PDE1B-related disorder or a dopamine D1 receptor intracellular signaling pathway disorder, by administering a modulator of PDE1B, preferably, but not limited to, an inhibitor of PDE1B or an agent that decreases the production of PDE1B.

What is claimed is:  
     
         1 . A method for modulating PDE1B activity in a cell or tissue comprising contacting said cell or tissue with an amount of a compound sufficient to modulate activity of a dopamine D1 intracellular signaling pathway, wherein contact of said cell or tissue with the compound results in modulation of PDE1B activity.  
     
     
         2 . The method of  claim 1  wherein the compound binds to PDE1B.  
     
     
         3 . The method of  claim 1  wherein the compound alters expression of PDE1B.  
     
     
         4 . The method of  claim 1  wherein phosphorylation of phospho-Thr34 of DARPP-32 (hereinafter “phospho-Thr34”) or phospho-Ser845 of GluR1 AMPA receptor (hereinafter “phospho-Ser845”) is modulated.  
     
     
         5 . A method of identifying a compound that modulates PDE1B activity in a dopamine D1 receptor intracellular signaling pathway in a cell or tissue comprising: 
 (a) determining a first level of PDE1B activity in said cell or tissue;    (b) contacting said cell or tissue with a test compound; and    (c) determining a second level of PDE1B activity in said cell or tissue,    wherein a difference in said first level and said second level of PDE1B activity is indicative of the ability of said test compound to modulate PDE1B activity.    
     
     
         6 . The method of  claim 5  wherein phosphorylation of phospho-Thr34 of DARPP-32 (hereinafter “phospho-Thr34”) or phospho-Ser845 of GluR1 AMPA receptor (hereinafter “phospho-Ser845”) is modulated.  
     
     
         7 . The method of  claim 5  wherein the difference in PDE1B activity is indicative of the ability of said test compound to modulate phosphorylation-dependent activation of DARPP-32 or GluR1 AMPA receptor.  
     
     
         8 . The method of  claim 5  wherein the difference in PDE1B activity is indicative of the ability of said test compound to modulate phosphorylation-dependent activation of ARPP-16, ARPP-19, ARPP-21, CREB, cAMP, cGMP, CK1, CK2, Cdk5, PKA, PKG, PP-2C, PP-2B, PP-1, calcium channels, Na/K ATPase or NMDA receptor.  
     
     
         9 . A method of identifying a compound that modulates activity of a dopamine D1 receptor intracellular signaling pathway in a cell or tissue comprising: 
 (a) determining a first level of activity of PDE1B in said cell or tissue;    (b) contacting said cell or tissue with a test compound; and    (c) determining a second level of activity of PDE1B in said cell or tissue,    wherein a difference in said first level and said second level of PDE1B activity is indicative of the ability of said test compound to modulate activity of said pathway.    
     
     
         10 . The method of  claim 9  wherein phosphorylation of phospho-Thr34 of DARPP-32 (hereinafter “phospho-Thr34”) or phospho-Ser845 of GluR1 AMPA receptor (hereinafter “phospho-Ser845”) is modulated.  
     
     
         11 . The method of  claim 9  wherein the difference in PDE1B activity is indicative of the ability of said test compound to modulate phosphorylation-dependent activation of DARPP-32 or GluR1 AMPA receptor.  
     
     
         12 . The method of  claim 9  wherein the difference in PDE1B activity is indicative of the ability of said test compound to modulate phosphorylation-dependent activation of ARPP-16, ARPP-19, ARPP-21, CREB, cAMP, cGMP, CK1, CK2, Cdk5, PKA, PKG, PP-2C, PP-2B, PP-1, calcium channels, Na/K ATPase or NMDA receptor.  
     
     
         13 . A method of identifying a compound that modulates activity of a dopamine D2 receptor, nitric oxide, noradrenergic, neurotensin, CCK, VIP, serotonin, glutamate, GABA, acetylcholine, adenosine, cannabinoid receptor, natriuretic peptide or endorphin intracellular signaling pathway in a cell or tissue comprising: 
 (a) determining a first level of activity of PDE1B in said cell or tissue;    (b) contacting said cell or tissue with a test compound; and    (c) determining a second level of activity of PDE1B in said cell or tissue,    wherein a difference in said first level and said second level of PDE1B activity is indicative of the ability of said test compound to modulate activity of said pathway.    
     
     
         14 . The method of  claim 13  wherein phosphorylation of phospho-Thr34 of DARPP-32 (hereinafter “phospho-Thr34”) or phospho-Ser845 of GluR1 AMPA receptor (hereinafter “phospho-Ser845”) is modulated.  
     
     
         15 . A method of identifying a compound that modulates activity of a dopamine D1 receptor intracellular signaling pathway in a cell or tissue comprising: 
 (a) contacting said cell or tissue with a test compound; and    (b) determining a level of activity of PDE1B in said cell or tissue;    wherein a difference in said level and a control level of PDE1B activity in a comparable cell or tissue not contacted with the test compound is indicative of the ability of said test compound to modulate activity of said pathway.    
     
     
         16 . The method of  claim 15  wherein the difference in said level and said control level of PDE1B activity is indicative of the ability of said test compound to modulate phosphorylation-dependent activation of DARPP-32 or GluR1 AMPA receptor.  
     
     
         17 . The method of  claim 15  wherein the difference in PDE1B activity is indicative of the ability of said test compound to modulate phosphorylation-dependent activation of ARPP-16, ARPP-19, ARPP-21, CREB, cAMP, cGMP, CK1, CK2, Cdk5, PKA, PKG, PP-2C, PP-2B, PP-1, calcium channels, Na/K ATPase or NMDA receptor.  
     
     
         18 . The method of  claim 15  wherein the difference in said level and said control level of PDE1B activity is indicative of the ability of said test compound to modulate activity of a dopamine D2 receptor, nitric oxide, noradrenergic, neurotensin, CCK, VIP, serotonin, glutamate, GABA, acetylcholine, adenosine, cannabinoid receptor, natriuretic peptide or endorphin intracellular signaling pathway.  
     
     
         19 . A method of identifying a compound that modulates PDE1B activity in a dopamine D1 receptor intracellular signaling pathway in a cell or tissue comprising: 
 (a) contacting said cell or tissue with a test compound; and    (b) determining a level of PDE1B activity in said cell or tissue;    wherein a difference in said level and a control level of PDE1B activity in a comparable cell or tissue not contacted with the test compound is indicative of the ability of said test compound to modulate PDE1B activity.    
     
     
         20 . The method of  claim 19  wherein the difference in said level and said control level of PDE1B activity is indicative of the ability of said test compound to modulate phosphorylation-dependent activation of DARPP-32 or GluR1 AMPA receptor.  
     
     
         21 . The method of  claim 19  wherein the difference in PDE1B activity is indicative of the ability of said test compound to modulate phosphorylation-dependent activation of ARPP-16, ARPP-19, ARPP-21, CREB, cAMP, cGMP, CK1, CK2, Cdk5, PKA, PKG, PP-2C, PP-2B, PP-1, calcium channels, Na/K ATPase or NMDA receptor.  
     
     
         22 . The method of  claim 19  wherein the difference in said level and said control level of PDE1B activity is indicative of the ability of said test compound to modulate activity of a dopamine D1 receptor, dopamine D2 receptor, nitric oxide, noradrenergic, neurotensin, CCK, VIP, serotonin, glutamate, GABA, acetylcholine, adenosine, cannabinoid receptor, natriuretic peptide or endorphin intracellular signaling pathway.  
     
     
         23 . A method for identifying an agent to be tested for an ability to treat a PDE1B-related or dopamine D1 receptor intracellular signaling pathway disorder in a patient in need of such treatment comprising: 
 (a) contacting in a cell or tissue PDE1B and Thr34-dephosphorylated DARPP-32 with a potential agent; and    (b) detecting the amount of phosphorylation of Thr34-dephosphorylated DARPP-32,    wherein the agent is identified if an increase in the phosphorylation of Thr34-dephosphorylated DARPP-32 is detected in the presence of the potential agent.    
     
     
         24 . The method of  claim 23  wherein the ability to treat the PDE1B-related or dopamine D1 receptor intracellular signaling pathway disorder is tested.  
     
     
         25 . A method for identifying an agent to be tested for an ability to treat a PDE1B-related or dopamine D1 receptor intracellular signaling pathway disorder in a patient in need of such treatment comprising: 
 (a) contacting in a cell or tissue PDE1B and Ser845-dephosphorylated GluR1AMPA receptor with a potential agent; and    (b) detecting the amount of phosphorylation of Ser845-dephosphorylated GluR1AMPA receptor,    wherein the agent is identified if an increase in the phosphorylation of Ser845-dephosphorylated GluR1 AMPA receptor is detected in the presence of the potential agent.    
     
     
         26 . The method of  claim 25  wherein the ability to treat the PDE1B-related or dopamine D1 receptor intracellular signaling pathway disorder is tested.  
     
     
         27 . A method for identifying an agent to be tested for an ability to modulate activity of a dopamine D1 receptor intracellular signaling pathway in a cell or tissue comprising: 
 (a) determining a first level of PDE1B activity in said cell or tissue;    (b) contacting said cell or tissue with a potential agent; and    (c) determining a second level of PDE1B activity in said cell or tissue,    wherein a difference in said first level and said second level of PDE1B activity is indicative of the ability of said potential agent to modulate activity of the dopamine D1 receptor intracellular signaling pathway.    
     
     
         28 . The method of  claim 27  comprising the additional step of: 
 (d) determining whether the dopamine D1 receptor intracellular signaling pathway is modulated.  
 
     
     
         29 . A method for identifying an agent to be tested for an ability to modulate activity of a dopamine D2 receptor, nitric oxide, noradrenergic, neurotensin, CCK, VIP, serotonin, glutamate, GABA, acetylcholine, adenosine, cannabinoid receptor, natriuretic peptide or endorphin intracellular signaling pathway intracellular signaling pathway in a cell or tissue comprising: 
 (a) determining a first level of PDE1B activity in said cell or tissue;    (b) contacting said cell or tissue with a potential agent; and    (c) determining a second level of PDE1B activity in said cell or tissue,    wherein a difference in said first level and said second level of PDE1B activity is indicative of the ability of said potential agent to modulate activity of said intracellular signaling pathway.    
     
     
         30 . The method of  claim 29  comprising the additional step of: 
 (d) determining whether said intracellular signaling pathway is modulated.  
 
     
     
         31 . A method for identifying an agent to be tested for an ability to modulate activity of a dopamine D1 receptor intracellular signaling pathway in a cell or tissue comprising: 
 (a) contacting said cell or tissue with a potential agent; and    (b) determining a level of PDE1B activity in said cell or tissue;    wherein a difference in said level and a control level of PD1E1B activity in a comparable cell or tissue not contacted with the test compound is indicative of the ability of said test compound to modulate activity of the dopamine D1 receptor intracellular signaling pathway.    
     
     
         32 . The method of  claim 31  comprising the additional step of: 
 (c) determining whether activity of the dopamine D1 receptor intracellular signaling pathway is modulated.  
 
     
     
         33 . A method for identifying an agent to be tested for an ability to modulate activity of a dopamine D2 receptor, nitric oxide, noradrenergic, neurotensin, CCK, VIP, serotonin, glutamate, GABA, acetylcholine, adenosine, cannabinoid receptor, natriuretic peptide or endorphin intracellular signaling pathway intracellular signaling pathway in a cell or tissue comprising: 
 (a) contacting said cell or tissue with a potential agent; and    (b) determining a level of PDE1B activity in said cell or tissue,    wherein a difference in said level and a control level of PDE1B activity in a comparable cell or tissue not contacted with the test compound is indicative of the ability of said test compound to modulate activity of said intracellular signaling pathway.    
     
     
         34 . The method of  claim 33  comprising the additional step of: 
 (d) determining whether the activity of said intracellular signaling pathway is modulated.  
 
     
     
         35 . A method for selecting a potential therapeutic agent for use in the treatment of a PDE1B-related or dopamine D1 receptor intracellular signaling pathway disorder, comprising: 
 (a) administering a potential therapeutic agent to an animal;    (b) measuring the response of said animal to said potential therapeutic agent;    (c) comparing the response of said animal with that of a control animal to which the potential therapeutic agent has not been administered; and    (d) selecting a potential therapeutic agent based on the difference in responses observed between said animal and said control animal.    
     
     
         36 . The method of  claim 35  wherein the animal is a mouse.  
     
     
         37 . The method of  claim 35  wherein the disorder is Parkinson's disease.  
     
     
         38 . A method for selecting a potential therapeutic agent for use in the treatment of a PDE1B-related or dopamine D1 receptor intracellular signaling pathway disorder, comprising: 
 (a) administering a potential therapeutic agent to an animal;    (b) measuring the response of said animal to administration of a neurotransmitter that interacts with a dopamine D1 receptor intracellular signaling pathway;    (c) comparing the response of said animal with that of a control animal to which the potential therapeutic agent has not been administered; and    (d) selecting a potential therapeutic agent based on the difference in responses observed between said animal and said control animal.    
     
     
         39 . The method of  claim 38  wherein the animal is a mouse.  
     
     
         40 . The method of  claim 38  wherein the disorder is Parkinson's disease.  
     
     
         41 . A method for selecting a potential therapeutic agent for use in the treatment of a PDE1B-related or dopamine D1 receptor intracellular signaling pathway disorder, comprising: 
 (a) administering a potential therapeutic agent to an animal;    (b) measuring the response of said animal to dopamine administration;    (c) comparing the response to dopamine administration of said animal with that of a control animal to which the potential therapeutic agent has not been administered; and    (d) selecting a potential therapeutic agent based on the difference in responses observed between said animal and said control animal.    
     
     
         42 . The method of  claim 41  wherein the animal is a mouse.  
     
     
         43 . The method of  claim 41  wherein the disorder is Parkinson's disease.  
     
     
         44 . A method for selecting a potential therapeutic agent for use in the treatment of a PDE1B-related or dopamine D1 receptor intracellular signaling pathway disorder, comprising: 
 (a) administering a potential therapeutic agent to an animal;    (b) measuring the response of said animal, wherein the response is selected from the group consisting of: 
 (i) exhibition of exploratory behavior during a test period for horizontal locomotor activity;  
 (ii) exhibition of hyperactivity after administration of methamphetamine treatment;  
 (iii) path length in acquisition of a Morris water maze;  
 (iv) change in level of phospho-Thr34 or phospho-Ser845 in striatal slices from said animal; and  
 (v) change in level of phospho-Thr34 or phospho-Ser845 in nucleus accumbens slices from said animal,  
   (c) comparing the response of said animal with that of a control animal to which the potential therapeutic agent has not been administered; and    (d) selecting a potential therapeutic agent based on the difference in responses observed between said animal and said control animal.    
     
     
         45 . The method of  claim 44  wherein the animal is a mouse.  
     
     
         46 . The method of  claim 44  wherein the disorder is Parkinson's disease.  
     
     
         47 . A method for treating a PDE1B-related disorder or a dopamine D1 receptor intracellular signaling pathway disorder in a patient in need thereof, comprising administering to the patient an agent that modulates the activity of PDE1B, wherein PDE1B activity modulates phosphorylation of an intracellular signaling molecule.  
     
     
         48 . The method of  claim 47  wherein the intracellular signaling molecule is DARPP-32 or GluR1 AMPA receptor.  
     
     
         49 . The method of  claim 47  wherein the intracellular signaling molecule is ARPP-16, ARPP-19, ARPP-21, CREB, cAMP, cGMP, CK1, CK2, Cdk5, PKA, PKG, PP-2C, PP-2B, PP-1, calcium channels, Na/K ATPase or NMDA receptor.  
     
     
         50 . The method of  claim 47  wherein the agent promotes or increases the activity of PDE1B.  
     
     
         51 . The method of  claim 47  wherein the agent inhibits or decreases the activity of PDE1B.  
     
     
         52 . The method of  claim 47  wherein the disorder is Parkinson's disease.  
     
     
         53 . A method for treating a PDE1B-related disorder or a dopamine D1 receptor intracellular signaling pathway disorder in a patient by administering an amount of a compound of the invention sufficient to modulate PDE1B activity.  
     
     
         54 . The method of  claim 53  wherein the agent promotes or increases the activity of PDE1B.  
     
     
         55 . The method of  claim 53  wherein the agent inhibits or decreases the activity of PDE1B.  
     
     
         56 . The method of  claim 53  wherein the agent modulates activity of a dopamine D1 receptor intracellular signaling pathway via modulation of PDE1B activity.