2-{[4-氨基-5-(3,4,5-三甲氧基-苄基)-嘧啶-2-亚胺基]-甲基}-6-甲氧基-苯酚与酵母细胞和牛血清白蛋白的相互作用

Interaction of 2-{ [4-Amino-5-(3,4,5-trimethoxy-benzyl)-pyrimidin-2-ylimino]-methyl}-6-methoxy-phenol with S. pombe Cells and BSA

用等摩尔的邻香草醛(C8H8O3)与甲氧苄胺嘧啶(C14H18N4O3)反应,合成了一种新的化合物2-{[4-氨基-5-(3,4,5-三甲氧基-苄基)-嘧啶-2-亚胺基]-甲基}-6-甲氧基-苯酚(APYP).通过元素分析、核磁共振、摩尔电导、红外光谱和紫外光谱等手段对其进行了表征,其化学式为C22H24N4O5.采用微量热法测定了APYP在301.15 K时对粟酒裂殖酵母细胞的生长代谢的产热曲线,计算得到了粟酒裂殖酵母细胞生长速率常数k,抑制率I以及半抑制浓度IC50等热动力学参数.实验结果表明,随着APYP浓度的增加,粟酒裂殖酵母细胞的生长代谢速率常数k减小,抑制率I增加,半抑制浓度IC50为123.2 mg/L(或2.96×10-4mol/L),即APYP对粟酒裂殖酵母细胞有抑制作用.同时利用荧光光谱和紫外光谱等方法研究了APYP与牛血清白蛋白(BSA)在接近生理条件下的相互作用,探讨了荧光猝灭机理,计算了反应的结合常数、结合位点、结合距离及热力学参数,研究了APYP与BSA之间的能量转移情况;并通过同步荧光光谱考察了APYP对BSA二级结构的影响.

A new compound 2-{[4-amino-5-（3,4,5-trimethoxy-benzyl）-pyrimidin-2-ylimino]-methyl}-6-me-thoxy-phenol （ APYP ） was synthesized using equivalent moles of o-vanillin （ C8 H8 O3 ） and trimethoprim （ C14 H18 N4 O3 ） . Elemental analysis, molar conductivity, UV-Visual, IR and NMR spectroscopy were em-ployed to characterize the composition and structure of APYP. The analytical results showed that the molecular formula of the compound is C22 H24 N4 O5 . The thermokinetic properties of the action of APYP on growth meta-bolism of S. pombe were studied by a TAM air isothermal calorimeter at 301.15 K. The microbial growth rate constant（ k） , inhibition ratio（ I） and half inhibition concentration（ IC50 ） were obtained. Experimental results show that the k values of S. pombe decreased and I values of S. pombe increased with the increase in the con-centrations of APYP and the IC50 value of APYP was found to be 123.20 mg/L. The APYP has good inhibition activity on the growth of S. pombe. Besides, fluorescence spectroscopy and UV-Vis absorption spectroscopy were employed to investigate the interactions of APYP with bovine serum albumin（ BSA） under the approxima-tively physiological conditions. The binding constants, binding sites, binding distance, and the thermodynam-ic parameters were calculated. The energy transfer between the donor（ BSA） and acceptor（ APYP） was inves-tigated. Furthermore, synchronous fluorescence spectroscopy was used to investigate the structural change of BSA molecules with additions of APYP.