人参皂苷的结构修饰及其体外抗肿瘤活性的研究
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摘要
人参为五加科多年生草本植物人参(Panax ginseng C.A.Meyer)的干燥根及根茎，是我国的传统名贵药材。现代研究表明人参具有抗肿瘤作用，其主要药物活性成分为人参多糖和人参皂苷。随着研究的深入，发现人参皂苷及各单体化合物的抗肿瘤作用和机制较为广泛，可能是直接作用于肿瘤细胞，促进肿瘤细胞凋亡、分化以及增强机体免疫力的结果。由于人参皂苷元水溶性差、生物利用度低，影响了临床应用，故本文对人参皂苷元进行了结构修饰，期望改善水溶性，提高生物利用度，以筛选出活性较高的抗肿瘤药物。
本论文以人参根总皂苷为原料制备结构修饰底物。采用V(HCl)/V(50%EtOH)为10%，90℃水解4h的降解方法，得人参根总皂苷酸催化降解产物，经分离纯化得化合物1~化合物3。对其理化性质进行分析，确定化合物结构分别是：20(R)–人参二醇（化合物1）、20(R)–人参三醇（化合物2）和新人参二醇（化合物3）。单体人参皂苷元的获得是进行结构修饰研究的前提和基础。
本论文在前期人参皂苷结构修饰的基础上，根据人参皂苷的体内代谢机理，对化合物1~化合物3进行了琥珀酸结构修饰，分别得到了20(R)–人参二醇-3β-琥珀酸酯（化合物4）、20(R)–人参三醇-3β,6ɑ-二琥珀酸酯（化合物6）和新人参二醇-3β,12β-二琥珀酸酯（化合物8）。并分别转化成了相应的钠盐化合物，分别是20(R)–人参二醇-3β-琥珀酸钠（化合物5）、20(R)–人参三醇-3β,6ɑ-二琥珀酸钠（化合物7）和新人参二醇-3β,12β-二琥珀酸钠（化合物9）。并且，本论文对化合物1~化合物3进行了羟基的氧化结构修饰，分别得到了20(R)–3–酮–人参二醇（化合物10）、20(R)–3,6–二酮–人参三醇（化合物11）和3,12–二酮–新人参二醇（化合物12）。共计得到9个结构修饰产物，其中化合物8、化合物9和化合物12为新化合物。
本论文还对结构修饰产物进行了体外抗肿瘤活性的研究。采用MTT法检测结构修饰产物对人结肠癌细胞SW1116的增殖抑制作用，结果显示结构修饰产物对SW1116细胞增殖均有不同程度的抑制作用，其中化合物8、化合物9和化合物12的抑制作用较强。
本项研究为人参皂苷类抗肿瘤新药的进一步研制与开发奠定了坚实的理论基础，人参皂苷类抗肿瘤药物有较强的药用价值和广阔的应用前景。
Ginseng, the root of Panax ginseng C.A.Meyer, is a traditional valuable Chinese herbalmedicine. Modern researches showed that ginseng has the antitumor activity, and theprincipal active ingredients of the ginseng antitumor are the polysaccharides from panax andginsenoside. With the developing of the research, people found that the antitumor activityand the mechanism of ginsenoside are widespread. Its antitumor mechanism may be a resultof working on tumor cells, promoting tumor cells die and differentiation, and enhancing theantitumor immunity of the body. Monomer ginsenosides were deeply restricted in theirclinical application for their poor water solubility and low bioavailability in human body.The aim of this thesis is to semisyntheses ginsenosides derivatives to improve the watersolubility and bioavailability of ginsenosides.
The total ginsenoside from the root of Panax ginseng was used as initial reactant toprepare ginsenoside in this thesis. The total ginsenoside was hydrolyzed for4h at90℃in asolution of50%EtOH, which contained10%HCl. Three compounds have been isolatedfrom the acid hydrolysate. They were20(R)-panaxadiol (compound1),20(R)-panaxatriol(compound2) and new panaxadiol (compound3), respectively. Obtaining ginsengenin is thepremise and foundation of this experiment studying on structure modification.
This paper, which based on the previous researches on the structural modification ofginsenoside and the vivo metabolism mechanism, modified the structural of compound1,compound2and compound3. And we got three derivatives, which were20(R)-panax-adiol-3β-succinate (compound4),20(R)-panaxatriol-3β,6ɑ-two succinate (compound6) andnew panaxadiol-3β,12β-two succinate (compound8), respectively. They were transformedinto the corresponding sodium compounds. The experiment also studied the oxidativemodification of compound1,compound2and compound3. And we got three derivatives,which were20(R)-3-ketone-panaxadiol (compound10),20(R)-3,6-two ketone-panaxatriol(compound11) and3,12-two ketone-new panaxadiol(compound12). At last, nine structuremodified products were got.
This paper also studied the antitumor activity of the structure modified products. TheMTT method was used to exam the structure modified products to the tumor cell inhibitoryaction. The result showed that the structure modified products had the varying degree ofdamaging effect to the human colonic cancer cells SW1116. Compound8, compound9andcompound12, which were elected to be higher antitumor activity drugs, showed the stron-ger inhibition on human colon cancer cells SW1116.
This study laid a solid theoretical foundation for the further research and developmentof new antitumor drugs. The antitumor drugs from ginsenoside have high medicinal value and broad application prospect.
本论文以人参根总皂苷为原料制备结构修饰底物。采用V(HCl)/V(50%EtOH)为10%，90℃水解4h的降解方法，得人参根总皂苷酸催化降解产物，经分离纯化得化合物1~化合物3。对其理化性质进行分析，确定化合物结构分别是：20(R)–人参二醇（化合物1）、20(R)–人参三醇（化合物2）和新人参二醇（化合物3）。单体人参皂苷元的获得是进行结构修饰研究的前提和基础。
本论文在前期人参皂苷结构修饰的基础上，根据人参皂苷的体内代谢机理，对化合物1~化合物3进行了琥珀酸结构修饰，分别得到了20(R)–人参二醇-3β-琥珀酸酯（化合物4）、20(R)–人参三醇-3β,6ɑ-二琥珀酸酯（化合物6）和新人参二醇-3β,12β-二琥珀酸酯（化合物8）。并分别转化成了相应的钠盐化合物，分别是20(R)–人参二醇-3β-琥珀酸钠（化合物5）、20(R)–人参三醇-3β,6ɑ-二琥珀酸钠（化合物7）和新人参二醇-3β,12β-二琥珀酸钠（化合物9）。并且，本论文对化合物1~化合物3进行了羟基的氧化结构修饰，分别得到了20(R)–3–酮–人参二醇（化合物10）、20(R)–3,6–二酮–人参三醇（化合物11）和3,12–二酮–新人参二醇（化合物12）。共计得到9个结构修饰产物，其中化合物8、化合物9和化合物12为新化合物。
本论文还对结构修饰产物进行了体外抗肿瘤活性的研究。采用MTT法检测结构修饰产物对人结肠癌细胞SW1116的增殖抑制作用，结果显示结构修饰产物对SW1116细胞增殖均有不同程度的抑制作用，其中化合物8、化合物9和化合物12的抑制作用较强。
本项研究为人参皂苷类抗肿瘤新药的进一步研制与开发奠定了坚实的理论基础，人参皂苷类抗肿瘤药物有较强的药用价值和广阔的应用前景。
Ginseng, the root of Panax ginseng C.A.Meyer, is a traditional valuable Chinese herbalmedicine. Modern researches showed that ginseng has the antitumor activity, and theprincipal active ingredients of the ginseng antitumor are the polysaccharides from panax andginsenoside. With the developing of the research, people found that the antitumor activityand the mechanism of ginsenoside are widespread. Its antitumor mechanism may be a resultof working on tumor cells, promoting tumor cells die and differentiation, and enhancing theantitumor immunity of the body. Monomer ginsenosides were deeply restricted in theirclinical application for their poor water solubility and low bioavailability in human body.The aim of this thesis is to semisyntheses ginsenosides derivatives to improve the watersolubility and bioavailability of ginsenosides.
The total ginsenoside from the root of Panax ginseng was used as initial reactant toprepare ginsenoside in this thesis. The total ginsenoside was hydrolyzed for4h at90℃in asolution of50%EtOH, which contained10%HCl. Three compounds have been isolatedfrom the acid hydrolysate. They were20(R)-panaxadiol (compound1),20(R)-panaxatriol(compound2) and new panaxadiol (compound3), respectively. Obtaining ginsengenin is thepremise and foundation of this experiment studying on structure modification.
This paper, which based on the previous researches on the structural modification ofginsenoside and the vivo metabolism mechanism, modified the structural of compound1,compound2and compound3. And we got three derivatives, which were20(R)-panax-adiol-3β-succinate (compound4),20(R)-panaxatriol-3β,6ɑ-two succinate (compound6) andnew panaxadiol-3β,12β-two succinate (compound8), respectively. They were transformedinto the corresponding sodium compounds. The experiment also studied the oxidativemodification of compound1,compound2and compound3. And we got three derivatives,which were20(R)-3-ketone-panaxadiol (compound10),20(R)-3,6-two ketone-panaxatriol(compound11) and3,12-two ketone-new panaxadiol(compound12). At last, nine structuremodified products were got.
This paper also studied the antitumor activity of the structure modified products. TheMTT method was used to exam the structure modified products to the tumor cell inhibitoryaction. The result showed that the structure modified products had the varying degree ofdamaging effect to the human colonic cancer cells SW1116. Compound8, compound9andcompound12, which were elected to be higher antitumor activity drugs, showed the stron-ger inhibition on human colon cancer cells SW1116.
This study laid a solid theoretical foundation for the further research and developmentof new antitumor drugs. The antitumor drugs from ginsenoside have high medicinal value and broad application prospect.
引文
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