α-甲基查尔酮类衍生物的合成及抗宫颈癌活性研究

发布时间：2018-01-18 07:32

本文关键词：α-甲基查尔酮类衍生物的合成及抗宫颈癌活性研究　出处：《新疆医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:1.采用不同催化方法合成分离查尔酮类化合物异甘草素并分析各自优缺点。2.分析查尔酮类化合物异甘草素结构中α、β不饱和羰基在生物活性中的作用。3.设计合成及分离α-甲基查尔酮。4.对上述α-甲基查尔酮类衍生物分别评价其对抑制子宫颈癌细胞的细胞增殖和促进癌细胞凋亡作用。5.通过分子对接的方法对α-甲基查尔酮衍生物进行分子学上的研究。方法:1.利用经典Claisen-Schmidt反应,采用SOCl2/EtOH、KF/Al2O3配合超声及KOH/Al2O3配合微波三个催化条件合成查尔酮类异甘草素并进行结构表征。2.对SOCl2/EtOH、KF/Al2O3配合超声及KOH/Al2O3配合微波这三种催化条件合成异甘草素的产率、合成时间进行统计比较得出较优的合成条件。3.以相应的苯丙酮衍生物和苯甲醛衍生物为原料,以哌啶/无水乙醇为催化剂合成α-甲基查尔酮。4.以SiHa(人子宫颈鳞癌细胞)和HeLa(子宫颈癌细胞)细胞株作为体外模型,利用MTT法对目标化合物进行抗宫颈癌细胞增殖的活性的研究。5.以SiHa(人子宫颈鳞癌细胞)和HeLa(子宫颈癌细胞)细胞株作为体外模型利用流式细胞仪测定目标化合物对宫颈癌细胞促凋亡作用。6.运用分子对接技术将目标化合物与分子蛋白进行分子学上的研究。结果:1.以SOCl2/EtOH为催化剂合成异甘草素,产率在47.2%~80.6%之间,而反应产率对应的时间在1-4 h之间;KF/Al2O3配合超声为催化条件合成异甘草素,产率在12.1%-95.6%之间,然而产率的高低与其超声的时间跟超声的功率有关,最大产率是在超声功率为250 W的条件下超声30 min得到的;KOH/Al2O3配合微波为催化条件合成异甘草素,其结构经1H-NMR、13C-NMR谱确认,产率在10.3%-93.5%之间。同样,产率与微波的时间及微波功率有关,但并不是功率越高产率也越大,在微波功率150 W,微波时间为100 s时反应的产率最高。2.以相应的苯丙酮衍生物和苯甲醛衍生物为原料,以哌啶/无水乙醇为催化剂合成15个α-甲基查尔酮衍生物,其结构1H-NMR确认,产率在30.2%-65.9%之间。3.对合成的15个α-甲基查尔酮衍生物进行抗宫颈癌活性研究,利用MTT法考察目标化合物的抗宫颈癌细胞增殖的活性研究,其中IC50最低的是化合物3(0.035μM),(E)-1-(2,4-二羟苯基)-3-(4-二甲氨苯基)-2-甲基丙烯酮,对宫颈癌SiHa细胞的抑制活性。4.化合物3的浓度为1μg.ml-1、2μg.ml-1、4μg.ml-1时对宫颈癌SiHa的早期凋亡率分别为11.5%、32.5%、38.2%,对宫颈癌HeLa细胞的早期凋亡率分别为11.8%、31.2%、43.8%。5.将目标化合物3与蛋白质(PDB:3E47)进行分子对接,化合物3的酚羟基和羰基氧可以和20S Proteasome结合口袋内的多个氨基酸残基形成氢键作用。参与形成氢键的残基包括THR1、THR21、GLY47和SER129。同时我们测定了各氢键所在重原子间的距离,发现所形成的氢键中有4个为较强氢键(距离在2.5-3.0?范围内)。这些氢键作用是药化合物3和20S Proteasome蛋白的结合的主要驱动力之一。结论:通过三中不同的催化条件合成甘草查尔酮类化合物,并且对其目标化合物进行结构鉴定;α-甲基查尔酮类化合物对Si Ha细胞和HeLa细胞表现出较显著的抑制活性和促进凋亡作用;得到较强的抑制宫颈癌细胞增殖和促宫颈癌细胞凋亡作用的化合物3。并从分子学的角度解释了α-甲基查尔酮类化合物显著的抑制有丝分裂的作用。此结果对从甘草查尔酮异甘草素类衍生物中筛选出具有抗子宫颈癌的有效候选药物奠定重要的基础。
[Abstract]:Objective: using 1. different catalytic synthesis methods for separation of chalcones isoliquiritigenin and.2. analysis to analyze the advantages and disadvantages of chalcones isoliquiritigenin structure alpha, beta unsaturated carbonyl in the biological activity of the role of.3. synthesis and separation of alpha methyl chalcone.4. of the alpha methyl chalcone derivatives were evaluated the inhibition of cervical cancer cell proliferation and promote apoptosis of cancer cell.5. by molecular docking method of alpha methyl chalcone derivatives for molecular research. Methods: 1. by using the classical Claisen-Schmidt reaction, using SOCl2/EtOH, KF/Al2O3 and KOH/Al2O3 combined with ultrasonic microwave combined with three catalytic conditions for synthesis of chalcones and isoliquiritigenin structural characterization of.2. on SOCl2/EtOH, KF/Al2O3 and KOH/Al2O3 combined with ultrasonic microwave yield the three catalytic conditions of synthesis of isoliquiritigenin synthesis. The time for statistical comparisons between.3. synthesis was optimized with benzene acetone derivatives and corresponding benzaldehyde derivatives as raw materials, piperidine / ethanol as catalyst for synthesizing alpha methyl chalcone.4. in SiHa (human cervical squamous cell carcinoma (HeLa) and cervical cancer cells) cells as in vitro model,.5. study anti cervical cancer cell proliferation of target compounds by MTT method with SiHa (activity of human cervical squamous cell carcinoma (HeLa) and cervical cancer cells) cells as an in vitro model using flow cytometry on cervical cancer cell apoptosis.6. using molecular docking technology will target compounds and molecular protein molecular research the target compounds. Results: 1. to SOCl2/EtOH for the synthesis of isoliquiritin catalyst, yield between 47.2%~80.6%, and the reaction yield corresponding time in 1-4 h; KF/Al2O3 combined with ultrasonic catalytic The synthesis conditions of isoliquiritigenin, yield between 12.1%-95.6%, but the power yield and ultrasonic time and ultrasonic, the maximum yield is 250 W under the condition of ultrasound in 30 min ultrasonic power; KOH/Al2O3 combined with microwave catalytic conditions for synthesis of isoliquiritigenin, characterized by 1H-NMR, 13C-NMR spectra confirmed that the yield of 10.3%-93.5%. Also, the time and the yield of microwave power and microwave power, but not the more productive rate is larger, the microwave power 150 W, microwave time is 100 s when the highest yield of.2. reaction with benzene acetone derivatives and corresponding benzaldehyde derivatives as raw material, anhydrous ethanol / piperidine as catalyst synthesis of 15 alpha methyl chalcone derivatives, the structure of 1H-NMR confirmed that the yield of.3. was studied in 15 cervical cancer activity of anti alpha synthesis of methyl chalcone derivatives at 30.2%-65.9%, examined using MTT method Anti cervical cancer cells proliferation activity of the target compounds, of which IC50 is the lowest of 3 compounds (0.035 M), (E) -1- (2,4- dihydroxy phenyl) -3- (4- two ammonia nitrogen phenyl) -2- methyl acrylic ketone compounds, the inhibitory activity of.4. on human cervical cancer cell line SiHa 3 concentration of 1 g.ml-1,2 g.ml-1,4 g.ml-1 of SiHa cervical cancer and early apoptosis rate were 11.5%, 32.5%, 38.2%, on cervical cancer HeLa cell early apoptosis rate were 11.8%, 31.2%, 43.8%.5. to 3 target compounds and protein (PDB:3E47) by molecular docking, 3 compounds of phenol hydroxyl and carbonyl oxygen and 20S combined with Proteasome in the pocket of a plurality of amino acid residues to form hydrogen bonds. Residues involved in the formation of hydrogen bonds including THR1, THR21, GLY47 and SER129. at the same time, we measured the distance between the heavy atoms of hydrogen bonds, found by the formation of hydrogen bonds in 4 for the strong hydrogen bonds (the distance in 2.5-3.0? Range). These hydrogen bonding is one of the main driving force of drug compounds with 3 and 20S Proteasome protein. Conclusion: the catalytic conditions of three different synthetic licorice chalcone compounds, and identify the structure of the target compound; alpha methyl chalcone compounds on Si Ha cells and HeLa cells showed inhibition the activity was significantly and apoptosis; proliferation of cervical cancer cells by compound strong and promote cervical cancer cell apoptosis and 3. from molecular explanation for the alpha methyl chalcone compounds significantly inhibit the mitogenic effect. The results of the Licochalcone isoliquiritigenin derivatives in screening the foundation of effective drug candidate has against cervical cancer.

【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R914;R96

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