pH-sensitive modified chitosan transgene vector, preparation method and applications thereof
A transgenic vector, chitosan technology, applied in other methods of inserting foreign genetic materials, recombinant DNA technology, etc., can solve the problems of low biodegradability, poor biocompatibility, low transfection efficiency, etc., and achieve biocompatibility Good property, low cytotoxicity, and the effect of improving transfection efficiency
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Embodiment 1
[0037] Dimethylmaleic anhydride (5.86g, 46.5mmol, sigma) was dissolved in a mixed solution of N,N-dimethylformamide (48mL) and water (2mL), and chitosan (2.5g, 15.5 mmol of sugar ring structural unit, sigma), after mixing uniformly, the mixture was stirred and reacted at 130° C. for 5 hours in a nitrogen atmosphere. After the reaction, the product was cooled to room temperature, precipitated with ethanol and washed three times, and finally vacuum-dried to obtain chitosan modified with dimethyl maleic anhydride, which was named CS-DMA. 1 H-NMR analysis product structure: the degree of substitution of dimethyl maleic anhydride is about 95%.
[0038] The dimethyl maleic anhydride-modified chitosan obtained above (1.2 g, 5 mmol sugar ring structural unit) was dissolved in 1 M sodium hydroxide solution (100 mL), placed in an ice-water bath and stirred evenly. Then p-toluenesulfonyl chloride (7.62g, 40mmol, sigma) was dissolved in acetonitrile (40mL), added dropwise in the sodium h...
Embodiment 2
[0042] Dimethylmaleic anhydride (5.86g, 46.5mmol, sigma) was dissolved in a mixed solution of N,N-dimethylformamide (48mL) and water (2mL), and chitosan (2.5g, 15.5 mmol of sugar ring structural unit, sigma), after mixing uniformly, the mixture was stirred and reacted at 130° C. for 5 hours in a nitrogen atmosphere. After the reaction, the product was cooled to room temperature, precipitated with ethanol and washed three times, and finally vacuum-dried to obtain chitosan modified with dimethyl maleic anhydride, which was named CS-DMA. 1 H-NMR analysis product structure: the degree of substitution of dimethyl maleic anhydride is about 95%.
[0043] The dimethyl maleic anhydride-modified chitosan obtained above (1.2 g, 5 mmol sugar ring structural unit) was dissolved in 1 M sodium hydroxide solution (100 mL), placed in an ice-water bath and stirred evenly. Then p-toluenesulfonyl chloride (9.53g, 50mmol, sigma) was dissolved in acetonitrile (40mL), added dropwise in the sodium h...
Embodiment 3
[0047] Dimethylmaleic anhydride (5.86g, 46.5mmol, sigma) was dissolved in a mixed solution of N,N-dimethylformamide (48mL) and water (2mL), and chitosan (2.5g, 15.5 mmol of sugar ring structural unit, sigma), after mixing uniformly, the mixture was stirred and reacted at 130° C. for 5 hours in a nitrogen atmosphere. After the reaction, the product was cooled to room temperature, precipitated with ethanol and washed three times, and finally vacuum-dried to obtain chitosan modified with dimethyl maleic anhydride, which was named CS-DMA. 1 H-NMR analysis product structure: the degree of substitution of dimethyl maleic anhydride is about 95%.
[0048] The dimethyl maleic anhydride-modified chitosan obtained above (1.2 g, 5 mmol sugar ring structural unit) was dissolved in 1 M sodium hydroxide solution (100 mL), placed in an ice-water bath and stirred evenly. Then p-toluenesulfonyl chloride (14.29g, 75mmol, sigma) was dissolved in acetonitrile (40mL), added dropwise in the sodium ...
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