CN106074551B - 一种2-取代芳乙烯基-n-甲基化喹啉衍生物在制备抗肿瘤药物中的应用 - Google Patents
一种2-取代芳乙烯基-n-甲基化喹啉衍生物在制备抗肿瘤药物中的应用 Download PDFInfo
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Abstract
本发明提供了一种2‑取代芳乙烯基‑N‑甲基化喹啉衍生物在制备抗肿瘤药物中的应用，所述衍生物结构式如式I所示；其中，R为氢、五元或六元含氮杂环基；所述R中任意一个或多个氢被取代，取代基选自烷基、羟基、胺基。本发明所述2‑取代芳乙烯基‑N‑甲基化喹啉衍生物是一种新型的Top II催化型抑制剂，具有更低的毒副作用，其对抗肿瘤具有普适性。本发明所述衍生物对多种癌细胞株，具有显著的抑制作用，抑制活性在0.2‑16µM，绝大部分化合物的活性优于阳性对照VP‑16，在制备新型的抗肿瘤药物方面具有良好的应用前景。式I。
Description
技术领域
本发明属于药物与有机合成技术领域，更具体地，涉及一种2-取代芳乙烯基-N-甲基化喹啉衍生物在制备抗肿瘤药物中的应用。
背景技术
拓扑异构酶(Topisomerase,Top)II是一类在细胞内普遍存在，解决DNA在复制、转录、和染色体分离中产生的拓扑问题的酶，其作为一个重要的药物靶点在抗肿瘤药物研发中被广泛关注。
据研究，喹啉类生物碱具有多种生理活性。包括抗微生物、抗肿瘤、抗氧化等。二苯乙烯类化合物也同样具有良好的生物活性和低毒性，例如白藜芦醇就是这样一个例子，有鉴于此，我们喹啉环的2-位引入芳乙烯基，并在4-位引入不同的胺链考察对抗肿瘤活性的影响，研究他们在抗肿瘤方面的应用。
发明内容
本发明根据现有技术中的不足，提供了一种2-取代芳乙烯基-N-甲基化喹啉衍生物在制备抗肿瘤药物中的应用。
本发明的技术目的通过以下技术方案实现：
本发明提供了一种2-取代芳乙烯基-N-甲基化喹啉衍生物在制备抗肿瘤药物中的应用，所述衍生物结构式如式I所示，
其中，R为氢、五元或六元含氮杂环基；
所述R中任意一个或多个氢被取代，取代基选自烷基、羟基、胺基。
本发明在喹啉环的2-位引入芳乙烯基，并在4-位引入不同的胺链，同时发现当2-位引入N-乙基咔唑基，喹啉N季铵化后抗肿瘤活性得到大幅提高，因此作为一类活性较高的抗肿瘤物质。
优选地，R为氢、六元含氮杂环基或六元含氮氧杂环基；所述R中任意一个或多个氢被取代，取代基选自C1-5烷基、羟基、胺基。
优选地，R为氢、甲基哌嗪基、哌啶基、吗啉基、羟乙基哌嗪基、二甲氨基乙基哌嗪基或二甲氨基丙基哌嗪基。
优选地，所述抗肿瘤药物为抗宫颈癌、乳腺癌、肝癌或淋巴癌的药物。
优选地，所述药物剂型为注射剂、片剂、丸剂、胶囊剂、悬浮剂或乳剂。
本发明同时提供一种2-取代芳乙烯基-N-甲基化喹啉衍生物在制备抑制拓扑异构酶II活性的药物中的应用，所述衍生物结构式如式I所示，
其中，R为氢、五元或六元含氮杂环基；
所述R中任意一个或多个氢被取代，取代基选自烷基、羟基、胺基。
优选地，R为氢、六元含氮杂环基或六元含氮氧杂环基；
所述R中任意一个或多个氢被取代，取代基选自C1-5烷基、羟基、胺基。
优选地，R为氢、甲基哌嗪基、哌啶基、吗啉基、羟乙基哌嗪基、二甲氨基乙基哌嗪基或二甲氨基丙基哌嗪基。
本发明在喹啉环的2-位引入芳乙烯基，并在4-位引入不同的胺链，同时发现当2-位引入咔唑基是必不可少的，当咔唑基换成苯基或取代苯基，对Top II的抑制活性将降低。
本发明所述化合物对多种细胞株，具有显著的抑制作用，抑制活性在0.2-16μM，绝大部分化合物的活性优于阳性对照VP-16。
Top II抑制剂分为Top II毒剂和催化型抑制剂，前者抑制酶的活性通过与酶形成酶-药物-DNA三元复合物，促进DNA往形成断裂中间体的方向进行，由于断裂DNA会诱导机体对DNA进行修复和重组，可能会产生耐药性和基因毒性，因此Top II毒剂往往具有较大的毒性和副作用，代表性药物如VP-16。而Top II催化型抑制剂由于不形成断裂DNA因此具有更低的毒性和副作用，为目前Top II抑制剂研究的热点方向。我们的实验证明，本发明所公开的新型2-取代芳乙烯基-N-甲基化喹啉衍生物在体外实验中，SPS-L3在10μM时可以完全抑制Top II对超螺旋DNA的解旋作用，远高于阳性对照VP16(etoposide)的抑制浓度(50μM)。而且与传统的Top II毒剂不同，2-取代芳乙烯基-N-甲基化喹啉衍生物不会形成酶-DNA-药物三元复合物，不会产生断裂DNA。
与现有技术相比，本发明具有以下有益效果：
(1)所合成的系列化合物证明咔唑基是必不可少的，当咔唑基换成苯基或取代苯基，对Top II的抑制活性将降低。
(2)本发明提供的化合物对多种细胞株，具有显著的抑制作用，抑制活性在0.2-16μM，绝大部分化合物的活性优于阳性对照VP-16。在制备新型的抗肿瘤药物方面具有良好的应用前景。因此本发明所述的2-取代芳乙烯基-N-甲基化喹啉衍生物可用于制备抗癌的药物。
(3)本发明2-取代芳乙烯基-N-甲基化喹啉衍生物是一种新型的Top II催化型抑制剂，具有更低的毒副作用。另外，Top II在肿瘤细胞中普遍高表达，已成为肿瘤治疗的重要靶标，我们的化合物证明对Top II有效，说明其抗肿瘤具有普适性。
附图说明
图1.SPS-L3抑制Top II对pBR322超螺旋质粒DNA的解旋实验图。
图2.SPS-L3对Top II的断裂DNA实验影响图。
具体实施方式
下面通过说明书附图和实施例对本发明进行具体描述，有必要在此指出的是本实施例只用于对本发明进行进一步说明，但不能理解为对本发明保护范围的限制，该领域的技术熟练人员可以根据上述本发明的内容作出一些非本质的改进和调整。
除非特别说明，本发明采用的试剂、方法和设备为本技术领域常规试剂、方法和设备。
化合物的合成，参照专利黄志纾，古练权，刘真权等，一种2-取代芳乙烯基-N-甲基化喹啉衍生物的制备及其在抗阿尔兹海默症药物中的应用，申请号2013102680611。
实施例1:本专利所述2-取代芳乙烯基-N-甲基化喹啉衍生物对Top II ATPase抑制作用
因为我们目标是为了获得Top II的催化型抑制剂，如果用全酶筛选，无法区分所得到的化合物是催化型抑制剂还是毒剂，所以我们在筛选时采用重组的酶，仅仅是Top II的ATPase部分，去掉了和DNA结合的C端和中间的催化区，这样可以有效的排除形成酶-药物-DNA三元复合物机制的影响，筛选所得到的化合物再进行其他实验的验证和确证。
孔雀绿-磷钼酸铵比色法检测化合物对Top II ATPase抑制活性，选用384孔板直接配制15μL的筛选体系，依次加入ddH2O，5×ATPase reaction buffer，ATPase，待测化合物，ATP。振荡器上震荡30s，转移至37℃烘箱孵育一定时间(Top II ATPase孵育时间为45min)，每孔加入40μL现配的孔雀绿试剂和5μL34％柠檬酸钠，混匀，静置15min后使用酶标仪测定样品在620nm下的紫外吸光度(OD值)，根据OD值的变化计算得到待测化合物的ATPase抑制活性。其中，每次加入1μg的Top II ATPase，被测试化合物终浓度为100μM，ATP的终浓度为1mM。阳性对照为1，4-萘醌
结果如表1所示，只有芳基为N-乙基取代咔唑基的化合物(SPS-L1～L7)才显示明显的Top II ATPase抑制活性，100μM下，抑制活性66-86％，比阳性对照1,4-萘醌(100μM，62％抑制率)还要高。苯环或取代苯环，吲哚基的活性不佳，而喹啉环的4位氨基取代则有很大的基团宽容度，受取代胺基种类影响较小。
表1. 2-取代芳乙烯基-N-甲基化喹啉衍生物对TopII ATPase抑制活性
实施例2：本专利所述2-取代芳乙烯基-N-甲基化喹啉衍生物对Hela，MCF-7,HepG2实体瘤肿瘤细胞株(贴壁细胞)的的抑制作用
我们将初筛中对Top II ATPase抑制活性大于50％的化合物(SPS-L1～L7)进行细胞水平的活性验证，以MTT法测试化合物的抗肿瘤活性，选取的细胞株包括Hela,MCF-7,HepG2实体瘤细胞株。取对数生长期的相应细胞株的细胞悬液(经0.25％胰蛋白酶消化使细胞脱落)0.5-1.0×105/ml,分装于96孔培养板中，150μL/孔，培养24小时，待细胞贴壁后，分别加入50μL相应不同浓度的药物或试液，实验设阴性对照组(生理盐水)、阳性对照组(VP16)和6个不同浓度给药组。每组设4个平行孔。置恒温5％CO2培养箱37℃培养48小时，在实验结束前4小时每孔加入20μL MTT液(5mg/ml)，4小时后吸弃培养液，每孔加入0.1mlDMSO，平板摇床震摇。5分钟待结晶溶解后置酶联检测仪，于570nm波长测各孔的OD值，按下列公式求生长抑制率，并用B1iss法求出半数抑制浓度IC50。
药物对肿瘤细胞抑制率＝(1-用药组平均OD值/对照组平均OD值)×100％。
结果如表2所示。所有化合物对三株肿瘤细胞Hela,MCF-7,HepG2显示很强的抗肿瘤活性，优于阳性对照VP16。
表2.化合物SPS-L1～L7对Hela,MCF-7,HepG2和CA-46肿瘤细胞的IC50(MTT,μM)
实施例3：本专利所述2-取代芳乙烯基-N-甲基化喹啉衍生物对CA-46肿瘤细胞株(悬浮细胞)的的抑制作用
取对数生长期的相应细胞株的细胞悬液0.5-1.0×105/ml,分装于96孔培养板中，150μL/孔，培养24小时，分别加入50μL相应不同浓度的药物或试液，实验设阴性对照组(培养基)、阳性对照组(VP16)和6个不同浓度给药组。每组设3个平行孔。置恒温5％CO2培养箱37℃培养48小时，在实验结束前4小时每孔加入20μL MTT液(2.5mg/ml)，4小时后每孔加入0.1ml三联液，平板摇床震摇。5分钟待结晶溶解后置酶联检测仪，于570nm波长测各孔的OD值，按下列公式求生长抑制率，并用B1iss法求出半数抑制浓度IC50。
药物对肿瘤细胞抑制率＝(1-用药组平均OD值/对照组平均OD值)×100％。
通过对体外细胞毒实验测试了化合物的抗肿瘤活性，结果如表2所示本专利所述化合物在体外对肿瘤细胞株CA-46具有较强的抑制作用。
实施例4：本专利所述2-取代芳乙烯基-N-甲基化喹啉衍生物对Top II抑制作用
在上述实施例2和3初步酶学水平和细胞水平筛选的基础上，选择对Top IIATPase有抑制活性的化合物对Top II全酶进行抑制活性实验，以确证化合物的对Top II的抑制活性。采用pBR322质粒松散法进行细胞体系外拓扑异构酶活性测定。其中重组人拓扑异构酶II购于TopoGEN公司。将1U的拓扑异构酶与待测药物(终浓度20，10，5，2.5，1.25，0.625μM)以及0.2μg超螺旋pBR322质粒混合加入拓扑异构酶缓冲液中，37℃水浴孵育30分钟后进行琼脂糖凝胶电泳，GelRed染色后利用凝胶成像仪进行检测。结果如图1所示，本专利所述的化合物在浓度为10μM时可以完全抑制Top II的活性，远高于阳性对照VP16(etoposide)的抑制浓度(50μM)。因此，本专利所述2-取代芳乙烯基-N-甲基化喹啉衍生物可用于制备以拓扑异构酶为靶点的抗癌药物。
实施例5:本专利所述2-取代芳乙烯基-N-甲基化喹啉衍生物对Top II介导的DNA断裂实验
该实验可以检测化合物抑制酶的活性过程是否有断裂DNA产生，是属于毒剂还是催化型抑制剂。
1.配制1％的琼脂糖凝胶及5×Top II buffer，随后使用ddH2O稀释化合物及配置0.1μg/μL的pBR322DNA溶液。
2.迅速从-80℃冰箱中取出购买回来的Topo II(浓度为20U/μL)并用ddH2O稀释得到浓度为5U/μL的Topo II溶液，置于冰上待用。
3.在20μL的反应体系中分别加入2μL的pBR322DNA溶液，2μL的Topo II溶液，2μL的化合物溶液，4μL的5×Topo II buffer，并用ddH2O补全体积。
4.配制好样品后，立即放入37℃水浴中孵育6min。
5.依次加入1μL 10％SDS，2μL 250mM NaEDTA(pH8.0)以及2μL 0.8mg/mL的蛋白酶K。45℃水浴中孵育30min。
6.样品与5μL 6×loading buffer混合后70℃水浴孵育2min。取12μL加入琼脂糖凝胶的样品孔中，65V电压下进行电泳1h。
7.电泳完毕后，将琼脂糖凝胶放入含有Gel-Red的1×TAE染色液中染色0.5h，再在65V电压下进行电泳1h，凝胶成像仪中拍照。
实验结果如图2显示，阳性对照VP16产生明显的缺口DNA(N)及线性DNA(L)，这都是DNA断裂的条带，而我们的化合物SPS-L3不仅无断裂条带的产生，而且还会减少VP16断裂条带的产生，本发明同时在除SPS-L3其他化合物中均发现此现象，说明该系列化合物是一典型的Top II催化型抑制剂。
Claims (3)
1.一种2-取代芳乙烯基-N-甲基化喹啉衍生物在制备抗肿瘤药物中的应用，其特征在于，所述衍生物结构式如式I所示，
式I；
其中，R为甲基哌嗪基、哌啶基、吗啉基、羟乙基哌嗪基、二甲氨基乙基哌嗪基或二甲氨基丙基哌嗪基。
2.根据权利要求1所述的应用，其特征在于，所述抗肿瘤药物为抗宫颈癌、乳腺癌、肝癌或淋巴癌的药物。
3.根据权利要求1所述的应用，其特征在于，所述药物的剂型为注射剂、片剂、丸剂、胶囊剂、悬浮剂或乳剂。
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101106992A (zh) * | 2005-01-18 | 2008-01-16 | 伊姆索公司 | 新颖的喹啉鎓盐和衍生物 |
| US20080064037A1 (en) * | 2006-03-09 | 2008-03-13 | New York University | Rna-selective probes for live cell imaging of nuclear structure and function |
| CN103333156A (zh) * | 2013-06-28 | 2013-10-02 | 中山大学 | 一种2-取代芳乙烯基-n-甲基化喹啉衍生物的制备及其在抗阿尔兹海默症药物中的应用 |
| CN104109125A (zh) * | 2014-06-26 | 2014-10-22 | 中山大学 | 一种4-取代-2-芳乙烯基喹啉类衍生物及其制备方法和应用 |
-
2016
- 2016-06-23 CN CN201610480497.0A patent/CN106074551B/zh active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101106992A (zh) * | 2005-01-18 | 2008-01-16 | 伊姆索公司 | 新颖的喹啉鎓盐和衍生物 |
| US20080064037A1 (en) * | 2006-03-09 | 2008-03-13 | New York University | Rna-selective probes for live cell imaging of nuclear structure and function |
| CN103333156A (zh) * | 2013-06-28 | 2013-10-02 | 中山大学 | 一种2-取代芳乙烯基-n-甲基化喹啉衍生物的制备及其在抗阿尔兹海默症药物中的应用 |
| CN104109125A (zh) * | 2014-06-26 | 2014-10-22 | 中山大学 | 一种4-取代-2-芳乙烯基喹啉类衍生物及其制备方法和应用 |
Non-Patent Citations (2)
| Title |
|---|
| Diversity-Oriented Fluorescence Library Approach (DOFLA) to the Discovery of Chymotrypsin Sensor;Shenliang Wang等;《J. Comb. Chem.》;20080315;第10卷;第460-465页 |
| Jie Dai等.Discovery of Small Molecules for Up-Regulating the Translation of Antiamyloidogenic Secretase, a Disintegrin and Metalloproteinase 10 (ADAM10), by Binding to the G‑Quadruplex-Forming Sequence in the 5′ Untranslated Region (UTR) of Its mRNA.《J. Med. Chem.》.2015,第58卷第3875-3891页. |
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