Preparation method of new type hyaluronic acid derivative containing coumarin fluorophore
A technology of sodium hyaluronate and its derivatives, which is applied in the field of biochemistry and can solve the problems that the synthesis of hyaluronic acid derivatives containing coumarin has not been reported in the literature.
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Embodiment 1
[0025] Add 0.1g (0.25mmol) sodium hyaluronate to a 25mL round bottom flask, then add 12mL dimethyl sulfoxide and 3mL water, stir with strong magnetic force to dissolve sodium hyaluronate, add 0.1M NaOH solution drop by drop to make the mixed solution pH=8-12, then add 0.173g (0.75mmol), 4-methyl-7-(2,3-epoxypropoxy)coumarin, and react at 30°C for 20h. The reaction mixture was put into a dialysis bag and dialyzed for 72 hours, and the dialysate was precipitated with ethanol to obtain a white solid, which was lyophilized and vacuum-dried for use in 1 Characterized by H NMR, by comparing 2.0ppm (3H, -NHCOCH 3 ) and 2.45ppm (3H, -CH on the introduced coumarin group 3) signal to calculate the degree of substitution of the product to be 1.8%.
Embodiment 2
[0027] Add 0.1g (0.25mmol) sodium hyaluronate to a 25mL round bottom flask, then add 12mL dimethyl sulfoxide and 3mL water, stir with strong magnetic force to dissolve sodium hyaluronate, add 0.1M NaOH solution drop by drop to make the mixed solution pH = 8-12, then add 0.173g (0.75mmol) coumarin derivative 2, and react at 30°C for 30h. The reaction mixture was put into a dialysis bag and dialyzed for 72 hours, and the dialysate was precipitated with ethanol to obtain a white solid, which was lyophilized and vacuum-dried for use in 1 Characterized by HNMR, by comparing 2.0ppm (3H, -NHCOCH 3 ) and 2.45ppm (3H, -CH on the introduced coumarin group 3 ) signal to calculate the degree of substitution of the product to be 2.4%.
Embodiment 3
[0029] Add 0.1g (0.25mmol) sodium hyaluronate to a 25mL round bottom flask, then add 12mL dimethyl sulfoxide and 3mL water, stir with strong magnetic force to dissolve sodium hyaluronate, add 0.1M NaOH solution drop by drop to make the mixed solution pH = 8-12, then add 0.173g (0.75mmol) coumarin derivative 2, and react at 30°C for 40h. The reaction mixture was put into a dialysis bag and dialyzed for 72 hours, and the dialysate was precipitated with ethanol to obtain a white solid, which was lyophilized and vacuum-dried for use in 1 Characterized by HNMR, by comparing 2.0ppm (3H, -NHCOCH 3 ) and 2.45ppm (3H, -CH on the introduced coumarin group 3 ) signal to calculate the degree of substitution of the product to be 3.6%.
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