本文主要研究内容
作者乔茜茜（2019）在《靶向载药多肽的设计及其多肽-阿霉素微球抗肿瘤活性分析》一文中研究指出：在药物研发中,学科交叉越发重要。合理设计多肽序列,使其作为高分子纳米材料,赋予其一定的靶向识别肿瘤细胞、包载传输药物和穿透细胞膜等能力具有重要意义。两亲性多肽纳米材料作为药物载体具有诸多优势:(1)提高难溶性药物溶解度。胶束疏水性内核能以共价键、疏水作用、范德华力等形式包载溶解性差的药物;由于这类药物在胶束的疏水核心中具有较高的分配系数,使其可以达到较高的载药量;(2)提高药物对环境的抗性。通过两亲性多肽链的包载,游离的药物进入人体内,可避免一些组织微环境因素的影响(例如一些酶会对蛋白类药物降解,吞噬小泡对小分子药物捕获、消噬);(3)提高人体对药物吸收率。两亲性多肽链的亲水段为其披上了一层“保护壳”,不易被吞噬小泡或内吞泡识别,延长了药物在代谢中的滞留时间,提高了人体对药物的吸收利用率;(4)靶向作用。通过引入一些具有特异性识别细胞表面受体的多肽序列,使载药胶束具有主动识别肿瘤位置的能力,提高疗效,减少不必要的药物浪费,降低毒害作用。本论文以固相合成技术为基础,设计和制备了具有靶向与载药功能的两亲性链状多肽P10(DGRGGGAAAA)。在此基础上,以阿霉素(DOX)为载药对象,制备靶向载药纳米微球。对P10进行红外光谱、高分辨率质谱等表征,以确定目标产物的合成,并达到进行体外细胞实验的纯度要求。对P10-DOX纳米微球的形态、基团结构、临界胶束浓度(CMC)以及体外释药性能等进行研究,为其进一步地应用提供理论依据。主要研究内容如下:1、采用固相合成法合成两亲性靶向多肽P10,优化其固相合成条件,确定的P10最佳合成条件为:以2-c1树脂为载体,N,N-二异丙基乙胺(DIEA)为活化试剂,TBTU为催化剂,氨基酸与树脂质量比为0.5:1,反应1.5 h。此条件下,粗肽产率达87.06%,纯度为95.55%;利用液质联用质谱仪对其分子量进行表征,高效液相色谱仪纯化后,P10纯度可达96.08%;采用紫外光谱和荧光光谱分析P10的CMC;研究结果表明,P10的CMC值约为0.45 mg/L;利用荧光光谱分析P10与DOX间的相互作用类型。分析结果显示,DOX对P10荧光具有较好的静态猝灭作用,相互作用力主要为氢键作用或范德华力。2、采用透析法装载DOX,将P10和DOX以质量比6:1混合后形成P10-DOX载药纳米微球。利用紫外分光光度法,构建DOX标准曲线及线性方程,并计算P10-DOX纳米微球的载药率(DLE)和包封率(EE).P10对DOX的EE和DLE值分别为23.011%±2.88%、10.125%±2.62%,说明P10对DOX具有良好的包载能力。利用红外光谱分析P10、DOX和P10-DOX的结构,通过对三种物质特征峰的比较,证明P10成功包载DOX;利用扫描电子显微镜(SEM)和Zeta粒度分析仪分别研究纳米微球的形态和大小,结果显示:P10-DOX复合物为大小均匀的球状胶束,粒径范围主要分布在390-530 nm。3、通过模拟胃液(pH1.2)、肠液(pH6.8)和肿瘤细胞环境(pH4.5)条件,研究P10-DOX能否在胃液、肠液和肿瘤环境中有效释药,结果显示:在前24 h内,药物会迅速释放到一定浓度,并在随后的72 h内稳定缓慢释放,达到药物缓释的效果。利用六个数学模型对P10-DOX的体外释药特性进行分析,分析结果表明P10-DOX纳米微球的体外释药特性符合Zero-order和Ritger-Peppas模型。利用MTT法考察P10对小鼠乳腺癌(4T1)细胞的细胞毒性,结果表明P10对4T1细胞无毒性。以Hela细胞为研究对象,考察P10-DOX体外抗肿瘤活性。结果表明:与DOX相比,P10-DOX对HaLa细胞的抑制率较高。当P10-DOX浓度为20 μg/mL时,肿瘤细胞抑制率为44.17%。4、采用固相合成法合成P13(DGRHHHLLLAAAA)肽,酸碱滴定法考察P13酸碱缓冲能力,FT-IR光谱分析各组分之间的连接情况,并用SEM对其表面形貌进行研究。通过对紫外光谱、荧光光谱、红外光谱、Zeta粒度分析仪以及体外释药等实验结果的分析表明:P13具有良好的酸碱缓冲能力,CMC值约为0.21 mg/L,P13-DOX纳米微球粒径的分布集中在122-164 nm:P13对DOX的EE和DLE值分别为20.40%±1.21%、21.25%±2.79%:不同pH条件下,P13-DOX纳米微球的体外释药特性均符合Weibull模型。最后,以4T1细胞为研究对象,考察其体外抗肿瘤活性。结果显示:当P13-DOX浓度为50μg/mL时,抑制率为26.65%。与P10-DOX相比,P13-DOX对肿瘤细胞的抑制率较高。当P13-DOX进入细胞后,首先定位在溶酶体中,2h后进入细胞核,并在细胞核中导致细胞凋亡。
Abstract
zai yao wu yan fa zhong ,xue ke jiao cha yue fa chong yao 。ge li she ji duo tai xu lie ,shi ji zuo wei gao fen zi na mi cai liao ,fu yu ji yi ding de ba xiang shi bie zhong liu xi bao 、bao zai chuan shu yao wu he chuan tou xi bao mo deng neng li ju you chong yao yi yi 。liang qin xing duo tai na mi cai liao zuo wei yao wu zai ti ju you zhu duo you shi :(1)di gao nan rong xing yao wu rong jie du 。jiao shu shu shui xing nei he neng yi gong jia jian 、shu shui zuo yong 、fan de hua li deng xing shi bao zai rong jie xing cha de yao wu ;you yu zhe lei yao wu zai jiao shu de shu shui he xin zhong ju you jiao gao de fen pei ji shu ,shi ji ke yi da dao jiao gao de zai yao liang ;(2)di gao yao wu dui huan jing de kang xing 。tong guo liang qin xing duo tai lian de bao zai ,you li de yao wu jin ru ren ti nei ,ke bi mian yi xie zu zhi wei huan jing yin su de ying xiang (li ru yi xie mei hui dui dan bai lei yao wu jiang jie ,tun shi xiao pao dui xiao fen zi yao wu bu huo 、xiao shi );(3)di gao ren ti dui yao wu xi shou lv 。liang qin xing duo tai lian de qin shui duan wei ji pi shang le yi ceng “bao hu ke ”,bu yi bei tun shi xiao pao huo nei tun pao shi bie ,yan chang le yao wu zai dai xie zhong de zhi liu shi jian ,di gao le ren ti dui yao wu de xi shou li yong lv ;(4)ba xiang zuo yong 。tong guo yin ru yi xie ju you te yi xing shi bie xi bao biao mian shou ti de duo tai xu lie ,shi zai yao jiao shu ju you zhu dong shi bie zhong liu wei zhi de neng li ,di gao liao xiao ,jian shao bu bi yao de yao wu lang fei ,jiang di du hai zuo yong 。ben lun wen yi gu xiang ge cheng ji shu wei ji chu ,she ji he zhi bei le ju you ba xiang yu zai yao gong neng de liang qin xing lian zhuang duo tai P10(DGRGGGAAAA)。zai ci ji chu shang ,yi a mei su (DOX)wei zai yao dui xiang ,zhi bei ba xiang zai yao na mi wei qiu 。dui P10jin hang gong wai guang pu 、gao fen bian lv zhi pu deng biao zheng ,yi que ding mu biao chan wu de ge cheng ,bing da dao jin hang ti wai xi bao shi yan de chun du yao qiu 。dui P10-DOXna mi wei qiu de xing tai 、ji tuan jie gou 、lin jie jiao shu nong du (CMC)yi ji ti wai shi yao xing neng deng jin hang yan jiu ,wei ji jin yi bu de ying yong di gong li lun yi ju 。zhu yao yan jiu nei rong ru xia :1、cai yong gu xiang ge cheng fa ge cheng liang qin xing ba xiang duo tai P10,you hua ji gu xiang ge cheng tiao jian ,que ding de P10zui jia ge cheng tiao jian wei :yi 2-c1shu zhi wei zai ti ,N,N-er yi bing ji yi an (DIEA)wei huo hua shi ji ,TBTUwei cui hua ji ,an ji suan yu shu zhi zhi liang bi wei 0.5:1,fan ying 1.5 h。ci tiao jian xia ,cu tai chan lv da 87.06%,chun du wei 95.55%;li yong ye zhi lian yong zhi pu yi dui ji fen zi liang jin hang biao zheng ,gao xiao ye xiang se pu yi chun hua hou ,P10chun du ke da 96.08%;cai yong zi wai guang pu he ying guang guang pu fen xi P10de CMC;yan jiu jie guo biao ming ,P10de CMCzhi yao wei 0.45 mg/L;li yong ying guang guang pu fen xi P10yu DOXjian de xiang hu zuo yong lei xing 。fen xi jie guo xian shi ,DOXdui P10ying guang ju you jiao hao de jing tai cu mie zuo yong ,xiang hu zuo yong li zhu yao wei qing jian zuo yong huo fan de hua li 。2、cai yong tou xi fa zhuang zai DOX,jiang P10he DOXyi zhi liang bi 6:1hun ge hou xing cheng P10-DOXzai yao na mi wei qiu 。li yong zi wai fen guang guang du fa ,gou jian DOXbiao zhun qu xian ji xian xing fang cheng ,bing ji suan P10-DOXna mi wei qiu de zai yao lv (DLE)he bao feng lv (EE).P10dui DOXde EEhe DLEzhi fen bie wei 23.011%±2.88%、10.125%±2.62%,shui ming P10dui DOXju you liang hao de bao zai neng li 。li yong gong wai guang pu fen xi P10、DOXhe P10-DOXde jie gou ,tong guo dui san chong wu zhi te zheng feng de bi jiao ,zheng ming P10cheng gong bao zai DOX;li yong sao miao dian zi xian wei jing (SEM)he Zetali du fen xi yi fen bie yan jiu na mi wei qiu de xing tai he da xiao ,jie guo xian shi :P10-DOXfu ge wu wei da xiao jun yun de qiu zhuang jiao shu ,li jing fan wei zhu yao fen bu zai 390-530 nm。3、tong guo mo ni wei ye (pH1.2)、chang ye (pH6.8)he zhong liu xi bao huan jing (pH4.5)tiao jian ,yan jiu P10-DOXneng fou zai wei ye 、chang ye he zhong liu huan jing zhong you xiao shi yao ,jie guo xian shi :zai qian 24 hnei ,yao wu hui xun su shi fang dao yi ding nong du ,bing zai sui hou de 72 hnei wen ding huan man shi fang ,da dao yao wu huan shi de xiao guo 。li yong liu ge shu xue mo xing dui P10-DOXde ti wai shi yao te xing jin hang fen xi ,fen xi jie guo biao ming P10-DOXna mi wei qiu de ti wai shi yao te xing fu ge Zero-orderhe Ritger-Peppasmo xing 。li yong MTTfa kao cha P10dui xiao shu ru xian ai (4T1)xi bao de xi bao du xing ,jie guo biao ming P10dui 4T1xi bao mo du xing 。yi Helaxi bao wei yan jiu dui xiang ,kao cha P10-DOXti wai kang zhong liu huo xing 。jie guo biao ming :yu DOXxiang bi ,P10-DOXdui HaLaxi bao de yi zhi lv jiao gao 。dang P10-DOXnong du wei 20 μg/mLshi ,zhong liu xi bao yi zhi lv wei 44.17%。4、cai yong gu xiang ge cheng fa ge cheng P13(DGRHHHLLLAAAA)tai ,suan jian di ding fa kao cha P13suan jian huan chong neng li ,FT-IRguang pu fen xi ge zu fen zhi jian de lian jie qing kuang ,bing yong SEMdui ji biao mian xing mao jin hang yan jiu 。tong guo dui zi wai guang pu 、ying guang guang pu 、gong wai guang pu 、Zetali du fen xi yi yi ji ti wai shi yao deng shi yan jie guo de fen xi biao ming :P13ju you liang hao de suan jian huan chong neng li ,CMCzhi yao wei 0.21 mg/L,P13-DOXna mi wei qiu li jing de fen bu ji zhong zai 122-164 nm:P13dui DOXde EEhe DLEzhi fen bie wei 20.40%±1.21%、21.25%±2.79%:bu tong pHtiao jian xia ,P13-DOXna mi wei qiu de ti wai shi yao te xing jun fu ge Weibullmo xing 。zui hou ,yi 4T1xi bao wei yan jiu dui xiang ,kao cha ji ti wai kang zhong liu huo xing 。jie guo xian shi :dang P13-DOXnong du wei 50μg/mLshi ,yi zhi lv wei 26.65%。yu P10-DOXxiang bi ,P13-DOXdui zhong liu xi bao de yi zhi lv jiao gao 。dang P13-DOXjin ru xi bao hou ,shou xian ding wei zai rong mei ti zhong ,2hhou jin ru xi bao he ,bing zai xi bao he zhong dao zhi xi bao diao wang 。
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论文作者分别是来自安徽工程大学的乔茜茜，发表于刊物安徽工程大学2019-07-18论文，是一篇关于两亲性多肽论文,阿霉素论文,肿瘤细胞论文,靶向论文,药物载体论文,逃逸能力论文,安徽工程大学2019-07-18论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自安徽工程大学2019-07-18论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：两亲性多肽论文; 阿霉素论文; 肿瘤细胞论文; 靶向论文; 药物载体论文; 逃逸能力论文; 安徽工程大学2019-07-18论文;