Method of Reducing Injury to Mammalian Cells

Abstract

A method of inhibiting the binding between N-methyl-D-aspartate receptors and neuronal proteins in a neuron is disclosed. The method comprises administering to the neuron an effective inhibiting amount of a peptide replacement agent for the NMDA receptor or neuronal protein interaction domain that effect said inhibition of the NMDA receptor-neuronal protein interaction. The method is of value in reducing the damaging effect of injury to mammalian cells. Postsynaptic density-95 protein (PSD-95) couples neuronal N-methyl-D-aspartate receptors (NMDARs) to pathways mediating excitotoxicity, ischemic and traumatic brain damage. This coupling was disrupted by transducing neurons with peptides that bind to modular domains on either side of the PSD-95/NMDAR interaction complex. This treatment attenuated downstream NMDAR signaling without blocking NMDAR activity, protected cultured cortical neurons from excitotoxic insults, dramatically reduced cerebral infarction volume in rats subjected to transient focal cerebral ischemia, and traumatic brain injury (TBI) in rats.

1 . A method of inhibiting the binding between a N-methyl-D-aspartate receptor and a neuronal protein in a neuron said method comprising administering to said neuron an effective inhibiting amount of a peptide replacement agent for the N-methyl-D-aspartate receptor or the neuronal protein to effect said inhibition of the binding between the N-methyl-D-aspartate receptor and the neuronal protein 
     
     
         2 . The method as defined in  claim 1  wherein said neuron is damaged. 
     
     
         3 . A method of reducing the damaging effect of ischemia or traumatic injury to the brain or spinal cord in a mammal, said method comprising treating said mammal with a non-toxic, damage-reducing, effective amount of a peptide replacement agent for a N-methyl-D-aspartate receptor or a neuronal protein to effect said inhibition of the binding between the N-methyl-D-aspartate receptor and the neuronal protein. 
     
     
         4 . The method as defined in  claim 3  wherein said mammal is under the influence of neuronal cell damage. 
     
     
         5 . The method as defined in  claim 3  wherein said N-methyl-D-aspartate receptor is bindable with a protein comprising at least one PDZ domain. 
     
     
         6 . The method as defined in  claim 5  wherein said protein comprising at least one PDZ domain is selected from the group consisting of PSD-95, PSD-93, SAP-102 and SAP-97. 
     
     
         7 . The method as defined in  claim 1  wherein said agent is a tSXV-containing peptide. 
     
     
         8 . The method as defined in  claim 7  wherein said agent is a SXV-containing peptide or a TXV-containing peptide. 
     
     
         9 . The method as defined in  claim 8  wherein said agent is KLSSIESDV [SEQ ID NO:1]. 
     
     
         10 . The method as defined in  claim 8  wherein said agent is KLSSIETDV [SEQ ID NO:2] 
     
     
         11 . The method as defined in  claim 8  wherein said agent further comprises a cell-membrane transduction domain. 
     
     
         12 . The method as defined in  claim 11  wherein said cell-membrane transduction domain is a human immunodeficiency virus type 1 (WV-1) Tat protein YORKKRRQRRR [SEQ ID NO:3]. 
     
     
         13 . The method as defined in  claim 11  wherein said agent comprises the amino acid sequence YGRKKRRQRRRKLSSIESDV [SEQ ID NO:4]. 
     
     
         14 . The method as defined in  claim 11  wherein said agent comprises the amino acid sequence YGRKKRRQRRRKLSSIETDV [SEQ ID NO:5]. 
     
     
         15 . The method as defined in  claim 11  wherein said cell-membrane transduction domain is an antennapedia internalization peptide. 
     
     
         16 - 28 . (canceled) 
     
     
         30 - 44 . A method of controlling the concentration of Ca 2+ -dependent signaling molecules in the vicinity of ion channel pores of cells in vivo to prevent the diffusion of toxic amounts of said Ca 2+  influx to prevent the triggering of neurotoxic phenomena, said method comprising administering an effective, nontoxic amount of a peptide replacement agent for a N-methyl-D-aspartate receptor or a neuronal protein to inhibit binding between the N-methyl-D-aspartate receptor and the neuronal protein. 
     
     
         30 - 45 . (canceled)