丹皮酚Schiff碱与氨基磷酸酯衍生物的合成研究
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摘要
丹皮酚是丹皮、徐长卿两味中药材的有效成分,具有抗心律失常,抗脉粥样硬化,抗肿瘤,增强免疫力和抗炎等多种药理作用,但是它的水溶性差、易挥发、不稳定、生物利用度低等因素又限制其疗效的发挥。对其进行结构修改以及作用机制等方面的探讨以期获得高效低毒、生物利用度高的丹皮酚衍生物,已经受到了国内外化学家和药物学家的重视。
本文根据药物设计的原理,考虑到氨基酸酯具有一定的生理活性,且Schiff碱具有一定的抗菌灭菌等作用,因此以丹皮酚为原料,与氨基酸酯在一定条件下反应得到了系列的丹皮酚氨基酸酯Schiff碱衍生物,并对反应溶剂等条件进行了优化；利用药物分子设计的前药原理,将具有一定活性哌嗪、氮芥等药效团通过磷酰化反应引入到丹皮酚中,合成了系列的含哌嗪等药效团的丹皮酚氨基磷酸酯衍生物；用荧光光谱法和紫外光谱法研究了部分化合物与牛血清蛋白的弱相互作用。具体研究内容如下：
首先对近几年来丹皮酚的含量测定方法、提取和分离方法、药理活性、结构修饰的研究进展进行了综述。
通过丹皮酚与氨基酸酯反应合成了8个的丹皮酚氨基酸酯Schiff碱衍生物,全部为新的化合物,其结构分别通过ESI MS、IR、NMR等进行了结构确认,部分化合物的结构进一步通过X-ray单晶衍射进行了确认。以丹皮酚与甘氨酸甲酯反应为模板,对反应溶剂、原料的物质的量比等条件进行了优化。
通过丹皮酚与含哌嗪、氮芥等药效团的磷酰化试剂反应,合成了8个新的丹皮酚氨基磷酸酯衍生物,其结构经ESI MS、IR、NMR等方法进行了鉴定,其中,通过2D NMR对化合物n的1H、13C NMR的数据进行了全归属。
利用荧光光谱法和紫外光谱法,对化合物i,j和k与牛血清蛋白的弱相互作用的情况进行了研究,结果表明：三种化合物都能使蛋白的荧光发生猝灭,且猝灭过程是由于形成复合物而引起的静态猝灭。
Pae, an active ingredient of two kinds of Chinese herbal medicines, Cynanchum and paniculatum, has anti-arrhythmic, anti-atherosclerotic vein, anti-tumor, anti-inflammatory, enhancing immunity, and other varieties of pharmacological effects. The related bioactivities of Pae have drawn more research attention of Chemists and pharmacologists at home and abroad. However its poor water-solubility, volatility, unstability and the related low bioavailability limit its bio application in some extents. So many works began to focus on the biofuntional mechanism study and the structure modification of Pae for obtaining relatively high efficiency, low toxicity and bioavailability of Pae derivatives. Taking into account the physiological activities of amino acid esters and at the same time the antibacterial activity of Schiff base, a series of new Pae Schiff derivatives were synthesized by additon and elimination reaction of carbonyl group of Pae with amino group of various amino acid esters under certain conditions. The related reaction conditions were optimized. Besides, a series of Paeonol amino phosphates of piperazine-containing pharmacophore were synthesized via sphosphorylation reaction, based on the prodrug principle of drug molecules design of introducing bioactive pharmacophore of piperazine and nitrogen mustard into the Pae. The bioacivity related non-covalent interactions of some target derivatives and bovine serum albumin were also investigated by fluorescence spectroscopy and UV spectra. The specific contents are as follows:
The dissertation first reviews the methods on Pae content determination, Pae extraction and isolation, Pae-related pharmacological activities and the structural modification in recent years. The template reaction using Paeonol and glycine methyl ester as reactants was then employed to optimize the related reaction conditions such as suitable solvent and molar ratios of raw materials etc. Then eight novel paeonol Schiff derivatives were synthesized under the optimized conditions and their structures were confirmed by ESI MS, IR, NMR. Moreover some structures of the new derivatives were further investigated and confirmed by X-ray diffraction. Following that, eight Paeonol amino phosphates of piperazine-containing pharmacophore were synthesized via sphosphorylation reaction of Pae with piperazine-containing, nitrogen mustard and other pharmacophore containing phosphoryl chloride. Their structures were confirmed by ESI MS, IR. NMR. The non-covalent interactions between three phosphorous containing derivatives and bovine serum albumin were investigated respectively using the fluorescence spectroscopy and ultraviolet spectroscopy. The result shows that, all of the three compounds can quench the fluorescence of protein, and the process is static quenching which is caused by complex formation.
本文根据药物设计的原理,考虑到氨基酸酯具有一定的生理活性,且Schiff碱具有一定的抗菌灭菌等作用,因此以丹皮酚为原料,与氨基酸酯在一定条件下反应得到了系列的丹皮酚氨基酸酯Schiff碱衍生物,并对反应溶剂等条件进行了优化；利用药物分子设计的前药原理,将具有一定活性哌嗪、氮芥等药效团通过磷酰化反应引入到丹皮酚中,合成了系列的含哌嗪等药效团的丹皮酚氨基磷酸酯衍生物；用荧光光谱法和紫外光谱法研究了部分化合物与牛血清蛋白的弱相互作用。具体研究内容如下：
首先对近几年来丹皮酚的含量测定方法、提取和分离方法、药理活性、结构修饰的研究进展进行了综述。
通过丹皮酚与氨基酸酯反应合成了8个的丹皮酚氨基酸酯Schiff碱衍生物,全部为新的化合物,其结构分别通过ESI MS、IR、NMR等进行了结构确认,部分化合物的结构进一步通过X-ray单晶衍射进行了确认。以丹皮酚与甘氨酸甲酯反应为模板,对反应溶剂、原料的物质的量比等条件进行了优化。
通过丹皮酚与含哌嗪、氮芥等药效团的磷酰化试剂反应,合成了8个新的丹皮酚氨基磷酸酯衍生物,其结构经ESI MS、IR、NMR等方法进行了鉴定,其中,通过2D NMR对化合物n的1H、13C NMR的数据进行了全归属。
利用荧光光谱法和紫外光谱法,对化合物i,j和k与牛血清蛋白的弱相互作用的情况进行了研究,结果表明：三种化合物都能使蛋白的荧光发生猝灭,且猝灭过程是由于形成复合物而引起的静态猝灭。
Pae, an active ingredient of two kinds of Chinese herbal medicines, Cynanchum and paniculatum, has anti-arrhythmic, anti-atherosclerotic vein, anti-tumor, anti-inflammatory, enhancing immunity, and other varieties of pharmacological effects. The related bioactivities of Pae have drawn more research attention of Chemists and pharmacologists at home and abroad. However its poor water-solubility, volatility, unstability and the related low bioavailability limit its bio application in some extents. So many works began to focus on the biofuntional mechanism study and the structure modification of Pae for obtaining relatively high efficiency, low toxicity and bioavailability of Pae derivatives. Taking into account the physiological activities of amino acid esters and at the same time the antibacterial activity of Schiff base, a series of new Pae Schiff derivatives were synthesized by additon and elimination reaction of carbonyl group of Pae with amino group of various amino acid esters under certain conditions. The related reaction conditions were optimized. Besides, a series of Paeonol amino phosphates of piperazine-containing pharmacophore were synthesized via sphosphorylation reaction, based on the prodrug principle of drug molecules design of introducing bioactive pharmacophore of piperazine and nitrogen mustard into the Pae. The bioacivity related non-covalent interactions of some target derivatives and bovine serum albumin were also investigated by fluorescence spectroscopy and UV spectra. The specific contents are as follows:
The dissertation first reviews the methods on Pae content determination, Pae extraction and isolation, Pae-related pharmacological activities and the structural modification in recent years. The template reaction using Paeonol and glycine methyl ester as reactants was then employed to optimize the related reaction conditions such as suitable solvent and molar ratios of raw materials etc. Then eight novel paeonol Schiff derivatives were synthesized under the optimized conditions and their structures were confirmed by ESI MS, IR, NMR. Moreover some structures of the new derivatives were further investigated and confirmed by X-ray diffraction. Following that, eight Paeonol amino phosphates of piperazine-containing pharmacophore were synthesized via sphosphorylation reaction of Pae with piperazine-containing, nitrogen mustard and other pharmacophore containing phosphoryl chloride. Their structures were confirmed by ESI MS, IR. NMR. The non-covalent interactions between three phosphorous containing derivatives and bovine serum albumin were investigated respectively using the fluorescence spectroscopy and ultraviolet spectroscopy. The result shows that, all of the three compounds can quench the fluorescence of protein, and the process is static quenching which is caused by complex formation.
引文
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