棉酚及其衍生物的制备及抗血吸虫病活性研究
发布时间：2019-05-09 12:26
【摘要】：血吸虫病是威胁人类健康的重大疾病之一,唯一有效的治疗药物是吡喹酮,大规模单一用药产生的耐药性血吸虫的出现,使开发新型抗血吸虫病药物的工作显得非常迫切。老药新用的策略可以提高抗血吸虫病药物的研发速度。文献报道,拥有广泛生物活性的棉酚具有抑制重组日本血吸虫的乳酸脱氢酶的活性,棉酚很有可能像青蒿素一样,具有抗日本血吸虫的杀虫活性。本文研究的基本思路是,通过提取棉仁中的棉酚,验证棉酚抗日本血吸虫的杀虫活性;通过化学合成获得多样性的棉酚衍生物化合物库,获得结构新颖、活性优越的抗日本血吸虫药物先导物。具体实施过程分以下两部分:1)以棉仁为原料,分别运用丙酮提取-苯胺沉淀法和碱液提取-苯胺沉淀法,通过评价两种提取方法获得的棉酚产率以及棉酚的纯度数据,获得新的棉酚提取工艺,提高棉酚的产量与质量;2)围绕棉酚的结构骨架,基于分子中的醛基,合成一系列棉酚衍生物,将得到的棉酚和棉酚衍生物进行体外抗血吸虫活性评价,获得构效关系信息,选取具有代表性的棉酚衍生物,进一步进行体内抗血吸虫活性研究,获得初步的体内治疗血吸虫的活性数据。研究结果显示,棉仁(20 g)经过丙酮提取得到棉酚粗提物(1 g,产率5%),获得的棉酚粗提物(1 g)利用苯胺沉淀法得到精制醋酸棉酚(400 mg,产率40%),纯度达到98%,提取到的棉酚质量占了棉仁总质量的2.0%,提取效果明显高于碱液提取-苯胺沉淀法。体外抗血吸虫活性结果显示,棉酚的杀虫活性优于药物吡喹酮,对雌虫的杀虫效果优于雄虫。合成的9个棉酚衍生物对血吸虫产生了不同程度的杀虫活性,构效关系结果显示,当R为取代苯环时,苯环上的氯原子取代不利于杀虫活性的提高,当R为苯环时,棉酚衍生物26的杀虫活性明显优于棉酚及其它取代苯环衍生物,当R为苄基或者环己基甲基时,杀虫活性明显低于苯基衍生物26,当苯丙氨酸甲酯与棉酚缩合产生衍生物29时,体外杀虫活性最高,在浓度10?M下,在48小时后,杀虫率达到100%,并且化合物29体外杀死雄虫的效果好于雌虫。进一步抗血吸虫童虫实验显示,棉酚衍生物29在浓度仅为2?M时,24小时、48小时及72小时,分别对童虫就显示了71.7%、75.0%以及81.7%的杀虫率,杀虫活性远远高于阳性对照青蒿甲酯和棉酚。最后,小鼠体内活性评价显示,通过腹腔给药,棉酚衍生物29对成虫的减虫率达到72.3%,对雌虫的减虫率达到76.8%,对虫卵的减卵率达到43.3%,棉酚衍生物29的体内杀虫活性和减卵率明显高于棉酚;通过灌胃给药,棉酚衍生物29的减虫率达到57.4%,减雌率达到53.5%,减卵率达到22.2%,体内杀虫效果高于棉酚。本研究发现新型抗日本血吸虫病药物并做出了全新的尝试,为发现新型抗血吸虫病药物提供新的研究思路。
[Abstract]:Schistosomiasis is one of the major diseases threatening human health. Praziquantel is the only effective therapeutic drug. The emergence of drug-resistant schistosomiasis produced by large-scale single drug makes the development of new anti-schistosomiasis drugs very urgent. The strategy of new use of old drugs can improve the research and development speed of anti-schistosomiasis drugs. It has been reported that gossypol, which has a wide range of biological activities, has the activity of inhibiting lactic dehydrogenase of recombinant Schistosoma japonicum. Gossypol, like artemisinin, has insecticidal activity against Schistosoma japonicum. The basic idea of this study is to verify the insecticidal activity of gossypol against Schistosoma japonicum by extracting gossypol from cotton kernel. A diverse library of gossypol derivatives was obtained by chemical synthesis, and a novel and active precursor of anti-Schistosoma japonicum (Schistosoma japonicum) was obtained. The implementation process is divided into the following two parts: 1) using cotton kernel as raw material, the yield of gossypol and the purity data of gossypol were evaluated by acetone extraction-Aniline precipitation method and Alkali extraction-Aniline precipitation method, respectively. A new extraction process of gossypol was obtained to improve the yield and quality of gossypol. 2) A series of gossypol derivatives were synthesized based on the aldehyde group in the molecule around the structural skeleton of gossypol. The antischistosomiasis activity of gossypol and gossypol derivatives in vitro was evaluated, and the structure-activity relationship information was obtained. Some representative gossypol derivatives were selected to study the anti-Schistosoma activity in vivo, and the preliminary data of anti-Schistosoma activity in vivo were obtained. The results showed that the crude extract of gossypol (1 g, yield 5%) was extracted from cotton kernel (20 g) by acetone, and the crude extract of gossypol (1 g) was obtained by phenylamine precipitation method. The yield of gossypol acetate was 40%, and the yield of gossypol acetate was 40%. The purity of gossypol reached 98%, and the extracted gossypol accounted for 2.0% of the total mass of cotton kernel. The extraction effect was obviously higher than that of alkali extraction-aniline precipitation. The results showed that gossypol had better insecticidal activity than praziquantel in vitro, and the insecticidal effect of gossypol against female was superior to that of male. Nine gossypol derivatives produced insecticidal activity against Schistosoma japonicum in varying degrees. The results of structure-activity relationship showed that the substitution of chlorine atoms on benzene ring was not conducive to the improvement of insecticidal activity when R was substituted for benzene ring, and when R was benzene ring, the structure-activity relationship showed that the substitution of chlorine atom on benzene ring was not conducive to the improvement of insecticidal activity. The insecticidal activity of gossypol derivative 26 is obviously better than that of gossypol and other substituted benzene ring derivatives. when R is benzyl or cyclohexyl methyl, the insecticidal activity of gossypol derivative 26 is significantly lower than that of phenyl derivative 26. when phenylalanine methyl ester is condensed with gossypol to produce derivative 29, The insecticidal activity of compound 29 was the highest in vitro. After 48 hours, the killing rate of compound 29 was 100%, and the killing effect of compound 29 was better than that of female. Further experiments against Schistosoma japonicum showed that gossypol derivative 29 showed 71.7%, 75.0% and 81.7% insecticidal rates against child insects at the concentration of only 2 脳 M for 24 hours, 48 hours and 72 hours, respectively, and the insecticidal rates of gossypol derivative 29 against Schistosoma japonicum were 71.7%, 75.0% and 81.7%, respectively. The insecticidal activity was much higher than that of Artemisia annua methyl ester and gossypol. Finally, the in vivo activity evaluation of mice showed that the worm reduction rate of gossypol derivatives 29 pairs of adults was 72.3%, that of female insects was 76.8%, and that of eggs was 43.3%. The insecticidal activity and egg reduction rate of gossypol derivative 29 were significantly higher than those of gossypol. The worm reduction rate of gossypol derivative 29 was 57.4%, the female reduction rate was 53.5%, and the egg reduction rate was 22.2%. The insecticidal effect of gossypol derivative 29 in vivo was higher than that of gossypol. This study found new anti-schistosomiasis drugs and made a new attempt to provide a new way of thinking for the discovery of new anti-schistosomiasis drugs.
【学位授予单位】：江南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R914;R96
[Abstract]:Schistosomiasis is one of the major diseases threatening human health. Praziquantel is the only effective therapeutic drug. The emergence of drug-resistant schistosomiasis produced by large-scale single drug makes the development of new anti-schistosomiasis drugs very urgent. The strategy of new use of old drugs can improve the research and development speed of anti-schistosomiasis drugs. It has been reported that gossypol, which has a wide range of biological activities, has the activity of inhibiting lactic dehydrogenase of recombinant Schistosoma japonicum. Gossypol, like artemisinin, has insecticidal activity against Schistosoma japonicum. The basic idea of this study is to verify the insecticidal activity of gossypol against Schistosoma japonicum by extracting gossypol from cotton kernel. A diverse library of gossypol derivatives was obtained by chemical synthesis, and a novel and active precursor of anti-Schistosoma japonicum (Schistosoma japonicum) was obtained. The implementation process is divided into the following two parts: 1) using cotton kernel as raw material, the yield of gossypol and the purity data of gossypol were evaluated by acetone extraction-Aniline precipitation method and Alkali extraction-Aniline precipitation method, respectively. A new extraction process of gossypol was obtained to improve the yield and quality of gossypol. 2) A series of gossypol derivatives were synthesized based on the aldehyde group in the molecule around the structural skeleton of gossypol. The antischistosomiasis activity of gossypol and gossypol derivatives in vitro was evaluated, and the structure-activity relationship information was obtained. Some representative gossypol derivatives were selected to study the anti-Schistosoma activity in vivo, and the preliminary data of anti-Schistosoma activity in vivo were obtained. The results showed that the crude extract of gossypol (1 g, yield 5%) was extracted from cotton kernel (20 g) by acetone, and the crude extract of gossypol (1 g) was obtained by phenylamine precipitation method. The yield of gossypol acetate was 40%, and the yield of gossypol acetate was 40%. The purity of gossypol reached 98%, and the extracted gossypol accounted for 2.0% of the total mass of cotton kernel. The extraction effect was obviously higher than that of alkali extraction-aniline precipitation. The results showed that gossypol had better insecticidal activity than praziquantel in vitro, and the insecticidal effect of gossypol against female was superior to that of male. Nine gossypol derivatives produced insecticidal activity against Schistosoma japonicum in varying degrees. The results of structure-activity relationship showed that the substitution of chlorine atoms on benzene ring was not conducive to the improvement of insecticidal activity when R was substituted for benzene ring, and when R was benzene ring, the structure-activity relationship showed that the substitution of chlorine atom on benzene ring was not conducive to the improvement of insecticidal activity. The insecticidal activity of gossypol derivative 26 is obviously better than that of gossypol and other substituted benzene ring derivatives. when R is benzyl or cyclohexyl methyl, the insecticidal activity of gossypol derivative 26 is significantly lower than that of phenyl derivative 26. when phenylalanine methyl ester is condensed with gossypol to produce derivative 29, The insecticidal activity of compound 29 was the highest in vitro. After 48 hours, the killing rate of compound 29 was 100%, and the killing effect of compound 29 was better than that of female. Further experiments against Schistosoma japonicum showed that gossypol derivative 29 showed 71.7%, 75.0% and 81.7% insecticidal rates against child insects at the concentration of only 2 脳 M for 24 hours, 48 hours and 72 hours, respectively, and the insecticidal rates of gossypol derivative 29 against Schistosoma japonicum were 71.7%, 75.0% and 81.7%, respectively. The insecticidal activity was much higher than that of Artemisia annua methyl ester and gossypol. Finally, the in vivo activity evaluation of mice showed that the worm reduction rate of gossypol derivatives 29 pairs of adults was 72.3%, that of female insects was 76.8%, and that of eggs was 43.3%. The insecticidal activity and egg reduction rate of gossypol derivative 29 were significantly higher than those of gossypol. The worm reduction rate of gossypol derivative 29 was 57.4%, the female reduction rate was 53.5%, and the egg reduction rate was 22.2%. The insecticidal effect of gossypol derivative 29 in vivo was higher than that of gossypol. This study found new anti-schistosomiasis drugs and made a new attempt to provide a new way of thinking for the discovery of new anti-schistosomiasis drugs.
【学位授予单位】：江南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R914;R96
【参考文献】
相关期刊论文 前10条
1 杨菲菲;汤鲁宏;;棉酚全合成及制剂研究进展[J];中国新药杂志;2016年14期
2 李玲;刘玉秀;汪清民;;棉酚的结构衍生及构效关系研究进展[J];中国科技论文;2015年12期
3 宋丽君;余传信;殷旭仁;沈双;高s，
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