Method and composition for protecting neuronal tissue from damage induced by elevated glutamate levels
a technology of elevated glutamate and neuronal tissue, which is applied in the direction of drug compositions, peptide/protein ingredients, cardiovascular disorders, etc., can solve the problems of neuronal death, excessive stimulation, and accelerated depletion of these limited energy resources, and achieve the effect of increasing the conversion of glutamate, enhancing pyruvate and oxaloacetate-mediated decreases in cellular or plasma glutamate concentrations
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example 1
In-Vitro Scavenging of Blood Glutamate by Activation of Resident Enzymes
[0178]Several intractable brain pathological conditions are characterized by the presence of excess glutamate in brain interstitial fluid. In order to increase the driving force for an enhanced brain-to-blood efflux of glutamate levels, conditions allowing decreased blood glutamate levels were determined in-vitro.
[0179]Since both glutamate pyruvate transaminase (GPT) and glutamate oxaloacetate transaminase (GOT) catalyze the degradation of glutamate (into 2-ketoglutarate—forward reaction) and its synthesis (from 2-ketoglutarate—reverse reaction) with equal efficiency, it was tested whether the administration of pyruvate and oxaloacetate to blood, results in a shifting of the equilibrium towards the degradation of glutamate i.e. the forward reaction. Materials and Experimental procedures
[0180]Experimental Procedures
[0181]Materials—Glutamate dehydrogenase was purchased from Roche (Roche diagnostics, GmBH, Mannheim...
example 2
Effects of Pyruvate and Oxaloacetate on Glutamate Levels in Different Blood Components
[0189]Whole blood can be divided into two major fractions, plasma and blood cells (i.e., erythrocytes, leukocytes and platelets). Since potentially therapeutic effects are proposed herein to be mediated via decreases in plasma glutamate concentration resulting in compensatory release from brain glutamate reserves, it is essential to determine glutamate levels in plasma and cellular pools following activation of endogenous GPT and GOT.
[0190]Results—Glutamate levels were therefore determined individually in blood cell fractions (closed symbols) and in plasma (open symbols), following repeated additions (arrows) of 1 mM pyruvate (squares), 1 mM oxaloacetate (triangles) or of a mixture of 1 mM pyruvate and 1 mM oxaloacetate (diamonds) (FIG. 2A). Glutamate determination was conducted as described in Example 1. The addition of pyruvate or oxaloacetate caused a comparable reduction in intracellular glutam...
example 3
Effective In Vitro Levels of Pyruvate and Oxaloacetate
[0192]Since previous examples have indicated that combined pyruvate and oxaloacetate administration is effective in lowering plasma and blood cell intracellular glutamate concentrations, it was essential to determine optimal concentrations of pyruvate and oxaloacetate for potential therapeutic application
[0193]Results—The extent of glutamate degradation reached after 60 minutes following the addition of increasing concentrations of pyruvate and oxaloacetate to plasma and blood cell fractions was determined. As shown in FIG. 3, half of the maximal effect was observed at a sub mM concentration in line with prior art Km values for pyruvate and oxaloacetate. Saturation was observed at a concentration of about 5 mM.
[0194]As both GPT and GOT utilize pyridoxal phosphate as a cofactor the addition of 15 μM pyridoxal phosphate was evaluated in terms of its ability to enhance pyruvate and oxaloacetate-mediated decreases in cellular or plas...
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