|Budget Amount *help
¥9,200,000 (Direct Cost : ¥9,200,000)
Fiscal Year 1994 : ¥1,900,000 (Direct Cost : ¥1,900,000)
Fiscal Year 1993 : ¥2,100,000 (Direct Cost : ¥2,100,000)
Fiscal Year 1992 : ¥5,200,000 (Direct Cost : ¥5,200,000)
1.Simple synthesis of inositol polyphosphates
Racemic 2,3-O-cyclohexylidene myoinositol was subjected to acetylation with acetic anhydride in the presence of Lipase CES in anhydrous dioxane. The hydroxyl groupe at C-3 position of L-enantiomer was acetylated to give L-1 and unacetylated D-1. D-1 is acetylated with acetic anhydride in DMF at C-6 and C-5 position. These were subjected to phosphorylation and deprotection procedure to afford D-Ins (1,4,5) P3and D-Ins (1,4,6) P3 in high yield. Starting from L-1, D-Ins (1,4,5,6) P4 and D- (1,2,4,5,6) P5 were obtained.
2.Preparation of inositol phosphate affinity column
p-Aminobenzoyl myo inositol derivatives were diazotizedy and then reacted with activated CH-Sepharose. Affinithy column of Ins (1,4,5) P3, Ins (1,3,4) P3, Ins (1,3,4,5) P4 were prepared.
3.Synthesis of membrane permiable Ins Px
In drder to penetrate the molecule through memblrane, minus charge of phosphoric acid must be diminished. Alkyl groups were substituted. But Ca was not released. Substituents must be hydrolized by enzyme in the cell. Protecting groups such as acetoxymethyl, p-acetoxybenzyl, phenoxyethyl are being tried to put on phosphoric acid group. Also large acyl group such palmitoyl is now on trial.
4.Syhthesis of PI (4,5) P2, and PI (3,4) P2 and their affinity column are now in progress.