Methods of suppressing microglial activation and systemic inflammatory responses

Abstract

Methods of suppressing the activation of microglial cells in the Central Nervous System (CNS), methods of ameliorating or treating the neurological effects of cerebral ischemia or cerebral inflammation, and methods of combating specific diseases that affect the CNS by administering a compound that binds to microglial receptors and prevents or reduces microglial activation are described. ApoE receptor binding peptides that may be used in the methods of the invention are also described, as are methods of using such peptides to treat peripheral inflammatory conditions such as sepsis. Also described are methods of screening compounds for the ability to suppress or reduce microglial activation.

That which is claimed is:  
     
         1 . A method of suppressing microglial activation, comprising exposing microglial cells to at least one compound that binds at a receptor bound by a peptide of ApoE in an amount effective to reduce microglial activation compared to that which would occur in the absence of the compound.  
     
     
         2 . The method of  claim 1 , wherein said at least one compound binds to a receptor bound by a peptide of ApoE having a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         3 . The method of  claim 1 , wherein said at least one compound is a receptor-binding ApoE peptide.  
     
     
         4 . The method of  claim 3 , wherein said ApoE peptide has a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         5 . A method of suppressing microglial activation in a mammalian subject, comprising administering to the brain of said subject a compound or a composition comprising a compound that binds to microglial cells at a receptor bound by a peptide of ApoE, wherein said compound or composition is administered in an amount effective to reduce microglial activation compared to that which would occur in the absence of the compound.  
     
     
         6 . The method of  claim 5 , wherein said compound binds to a receptor bound by a peptide of ApoE having a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         7 . The method of  claim 5 , wherein said compound is a receptor-binding ApoE peptide.  
     
     
         8 . The method of  claim 7 , wherein said ApoE peptide has a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         9 . A method of ameliorating symptoms associated with CNS inflammation in a subject, comprising administering to the brain of said subject a compound or a composition comprising a compound that binds to microglial cells at the receptor bound by a peptide of ApoE, said compound or composition is administered in an amount effective to reduce CNS inflammation compared to that which would occur in the absence of the compound.  
     
     
         10 . The method of  claim 9 , wherein said compound binds to a receptor bound by a peptide of ApoE having a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         11 . The method of  claim 9 , wherein said compound is a receptor-binding ApoE peptide.  
     
     
         12 . The method of  claim 11 , wherein said ApoE peptide has a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         13 . The method according to  claim 9 , wherein said subject is afflicted with a condition selected from the group consisting of Alzheimer's disease and multiple sclerosis.  
     
     
         14 . A method of ameliorating symptoms associated with CNS ischemia in a subject, comprising administering to the brain of said subject a compound or a composition comprising a compound that binds to microglial cells at the receptor bound by a peptide of ApoE, said compound or composition is administered in an amount effective to reduce CNS ischemic compared to that which would occur in the absence of the compound.  
     
     
         15 . The method of  claim 14 , wherein said compound binds to a receptor bound by a peptide of ApoE having a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         16 . The method of  claim 14 , wherein said compound is a receptor-binding ApoE peptide.  
     
     
         17 . The method of  claim 14 , wherein said ApoE peptide has a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         18 . A method according to  claim 14 , wherein said subject is afflicted with a condition selected from stroke, global cerebral ischemia, focal cerebral iscehmia, cerebral edema, closed acute head injury, and intracranial hemorrhage.  
     
     
         19 . A method of reducing neuronal cell injury associated with glutamate excitotoxicity in a mammalian subject by administering to said subject a compound or a composition comprising a compound that binds to microglial cells at the receptor bound by a peptide of ApoE, wherein said compound or composition is administered in an amount that reduces neuronal cell injury associated with glutamate excitotoxicity as compared to reduction that would occur in the absence of the compound.  
     
     
         20 . The method of  claim 19 , wherein said glutamate excitotoxicity is mediated by NMDA exposure.  
     
     
         21 . The method of  claim 19 , wherein said subject has been diagnosed with HIV dementia or encephalopy.  
     
     
         22 . The method of  claim 21 , wherein said compound is administered concurrently or sequentially with one or more HIV reverse transcriptase inhibitors or protease inhibitors.  
     
     
         23 . The method of  claim 19 , wherein said subject has been diagnosed with neurolathyrism, amyotrophic lateral sclerosis (ALS), Huntington's chorea, Parkinson's or schizophrenia.  
     
     
         24 . The method of  claim 23 , wherein said compound is administered concurrently or sequentially with one or more glutamate antagonists or antioxidizing substances.  
     
     
         25 . The method of  claim 24 , wherein said glutamate antagonist is Riluzole.  
     
     
         26 . The method of  claim 23 , wherein said subject has been diagnosed with Parkinson's disease.  
     
     
         27 . The method of  claim 26 , wherein said compound is administered concurrently or sequentially with one or more other compounds selected from the group consisting of NMDA receptor antagonists, MAO-B inhibitors and dopamine receptor antagonists.  
     
     
         28 . The method of  claim 27 , wherein said MAO-B inhibitor is L-deprenyl.  
     
     
         29 . The method of  claim 27 , wherein said dopamine receptor antagonist is Levodopera.  
     
     
         30 . The method of  claim 19 , wherein said subject has been diagnosed with bipolar disorder, epilepsy or d-2-hydroxyglutaric aciduria.  
     
     
         31 . The method of  claim 30 , wherein said compound is administered concurrently or sequentially with one or more anticonvulsant agents that inhibit NMDA receptor hypofunction.  
     
     
         32 . The method of  claim 32 , wherein said anticonvulsant agent is selected from the group consisting of phenytoin, carbamazepine, valproic acid, lamotrigine, riluzole, tetrodotoxin, felbamate, gabapentin and ethosuximide.  
     
     
         33 . The method of  claim 19 , wherein said subject has been diagnosed with multiple sclerosis or experimental allergic encephalomyelitis (EAE).  
     
     
         34 . The method of  claim 33 , wherein said compound is administered concurrently or sequentially with one or more compounds selected from the group consisting of interferon (IFN) beta-1b, IFN beta-1a, glatiramer acetate, IV immunoglobulin, methotrexate, azathioprine, memantine and other NMDA receptor antagonists.  
     
     
         35 . The method of  claim 19 , wherein said subject has suffered a traumatic brain injury.  
     
     
         36 . The method of  claim 19 , wherein said compound binds to a receptor bound by a peptide of ApoE having a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         37 . The method of  claim 19 , wherein said compound is a receptor-binding ApoE peptide.  
     
     
         38 . The method of  claim 37 , wherein said ApoE peptide has a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         39 . A method of anesthesizing a subject comprising administering to said subject a compound or a composition comprising a compound that binds to the peptide that is bound by a peptide of ApoE, wherein said compound or composition is administered in an amount effect to anesthesize said subject.  
     
     
         40 . The method of  claim 39 , wherein said compound binds to a receptor bound by a peptide of ApoE having a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         41 . The method of  claim 39 , wherein said compound is a receptor-binding ApoE peptide.  
     
     
         42 . The method of  claim 41 , wherein said ApoE peptide has a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         43 . A method of alleviating or reducing chronic pain in a subject comprising administering to said subject a compound or a composition comprising a compound that binds to a receptor that binds to a peptide of ApoE, wherein said compound or composition is administered in an amount that alleviates or reduces chronic pain in said patient.  
     
     
         44 . The method of  claim 43 , wherein said compound binds to a receptor bound by a peptide of ApoE having a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         45 . The method of  claim 43 , wherein said compound is a receptor-binding ApoE peptide.  
     
     
         46 . The method of  claim 45 , wherein said ApoE peptide has a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         47 . A method of alleviating, correcting or preventing substance abuse in a subject comprising administering to said subject a compound or a composition comprising a compound that binds to a receptor that binds to a peptide of ApoE, wherein said compound or composition is administered in an amount effective to alleviate, correct or prevent substance abuse in said subject.  
     
     
         48 . The method of  claim 47 , wherein said compound binds to a receptor bound by a peptide of ApoE having a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         49 . The method of  claim 47 , wherein said compound is a receptor-binding ApoE peptide.  
     
     
         50 . The method of  claim 49 , wherein said ApoE peptide has a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         51 . A method according to  claim 5  wherein said compound is conjugated to a carrier molecule, wherein the presence of the carrier molecule increases transport of said compound across the blood-brain barrier, compared to that which would occur in the absence of said carrier molecule.  
     
     
         52 . A method according to  claim 5  wherein the mode of administration is selected from parenteral administration, intrathecal administration, and spinal administration.  
     
     
         53 . A method according to  claim 5 , wherein said compound is administered prophylactically prior to scheduled surgery.  
     
     
         54 . A method of suppressing macrophage activation in a mammalian subject, by administering to said subject a compound or a composition comprising a compound that binds to macrophage cells at the receptor bound by a peptide of ApoE, wherein said compound or composition is administered in an amount that suppresses macrophage activation as compared to activation that would occur in the absence of the compound.  
     
     
         55 . The method of  claim 54 , wherein said compound binds to a receptor bound by a peptide of ApoE having a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         56 . The method of  claim 54 , wherein said compound is a receptor-binding ApoE peptide.  
     
     
         57 . The method of  claim 56 , wherein said ApoE peptide has a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         58 . A method of treating atherosclerosis or of reducing the formation of atherosclerotic plaques, comprising administering a compound or a composition containing a compound that binds to macrophage cells at the receptor bound by a peptide of ApoE, wherein the compound or composition is administered in an amount that reduces the formation of artherosclerotic plaques as compared to that which would occur in the absence of the compound.  
     
     
         59 . The method of  claim 58 , wherein said compound binds to a receptor bound by a peptide of ApoE having a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         60 . The method of  claim 58 , wherein said compound is a receptor-binding ApoE peptide.  
     
     
         61 . The method of  claim 60 , wherein said ApoE peptide has a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         62 . A method of treating or of reducing the inflammation associated with sepsis in a subject, comprising administering a compound or a composition containing a compound that binds to a receptor that binds to a peptide of ApoE, where said compound or composition is administered in an amount that reduces sepsis-associated inflammation as compared to that which would occur in the absence of the compound.  
     
     
         63 . The method of  claim 62 , wherein administration of said compound or composition results in a decrease in inflammatory cytokines as compared to that which would occur in the absence of said compound.  
     
     
         64 . The method of  claim 63 , wherein said inflammatory cytokines include TNFα or IL-6.  
     
     
         65 . The method of  claim 62 , wherein said compound is administered concurrently or sequentially with one or more anti-inflammatory cytokines or monoclonal antibodies.  
     
     
         66 . The method of  claim 65 , wherein said anti-inflammatory cytokines are selected from the group consisting of IL-10, transforming growth factor-beta, granulocyte colony-stimulating factor, IFN-phi, macrophage migration inhibitory factor and high mobility group 1 protein.  
     
     
         67 . The method of  claim 65 , wherein said monoclonal antibodies are selected from the group consisting of antiendotoxin antibodies, anti-tumor necrosis factor antibodies, and anti-CD14 antibodies.  
     
     
         68 . The method of  claim 62 , wherein said compound binds to a receptor bound by a peptide of ApoE having a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         69 . The method of  claim 62 , wherein said compound is a receptor-binding ApoE peptide.  
     
     
         70 . The method of  claim 69 , wherein said ApoE peptide has a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         71 . A therapeutic composition comprising a peptide selected from the group consisting of SEQ ID NO: 3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         72 . A method of screening a test compound for the ability to suppress glial or microglial activation, comprising 
 (a) incubating an activated glial or microglial cell culture with said compound, and    (b) measuring a marker of microglial activation,    wherein a decrease in microglial activation indicates that said compound has the ability to suppress microglial activation.    
     
     
         73 . The method of  claim 72 , wherein said marker of microglial activation is nitric oxide production.  
     
     
         74 . A method of screening a test compound for the ability to prevent or suppress glial or microglial activation, comprising: 
 (a) pre-incubating a glial or microglial cell culture with the compound;    (b) incubating the cell culture with a known activator of glia or microglia; and    (c) measuring a marker of glial or microglial activation,    wherein a decrease in or absence of microglial activation as compared to a culture not incubated with said compound indicates that said compound has the ability to suppress microglial activation.    
     
     
         75 . The method of  claim 74 , wherein said marker of microglial activation is nitric oxide production.  
     
     
         76 . A method of screening a test compound for the ability to suppress glial or microglial activation, comprising 
 (a) incubating a glial or microglial cell culture with the compound;    (b) incubating the cell culture with a receptor binding peptide of ApoE;    (c) determining whether the compound inhibits receptor binding of the peptide of ApoE by determining whether the compound inhibits the binding of the peptide to said glial or microglial cells,    thereby identifying a compound having the ability to suppress glial or microglial activation.    
     
     
         77 . The method of  claim 76 , wherein said peptide has a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:10.  
     
     
         78 . The method of  claim 76 , wherein said peptide binds to the LRP/α2M receptor.  
     
     
         79 . The method of  claim 76 , wherein said peptide is conjugated to or associated with a detectable label.  
     
     
         80 . The method of  claim 79 , wherein said label is selected from the group consisting of radioisotopes and fluorescent molecules.