CN113956292B - 一种具有聚集诱导发光性质的线粒体靶向化合物及其合成方法和应用 - Google Patents
一种具有聚集诱导发光性质的线粒体靶向化合物及其合成方法和应用 Download PDFInfo
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Abstract
本发明公开了一种具有聚集诱导发光性质的线粒体靶向化合物,该化合物通过将具有聚集诱导发光性质的荧光小分子与线粒体靶向基团结合,得到具有高抗癌活性和低毒副作用的产物;本发明化合物具有良好的细胞摄取能力和靶向肿瘤细胞内线粒体的能力;其次,本发明化合物通过抑制人源肿瘤细胞增殖、诱导癌细胞内活性氧产生、降低线粒体膜电位、改变线粒体形态、损伤线粒体功能以及诱导肿瘤细胞(线粒体)自噬,实现良好的抗肿瘤活性;由于本发明化合物的作用机制与金属抗肿瘤药物的作用机制不同,因此使用本发明化合物在抗肿瘤过程中不会产生耐药性的问题,从而保证持续性良好的抗癌效果;最后,本发明化合物能够有效抑制已对金属抗肿瘤药物(顺铂)产生耐药性的肿瘤细胞的增殖,从而解决现有化疗药物无法有效克服肿瘤细胞顺铂耐药性的问题。
Description
技术领域
本发明涉及一种具有聚集诱导发光性质的线粒体靶向化合物,还涉及上述化合物的合成方法和应用。
技术背景
癌症是全球范围内最致命的疾病之一。据世界卫生组织国际癌症研究机构(IARC)发布的2020年全球最新癌症数据显示,中国癌症新发人数远超世界其他国家。传统的肿瘤治疗包括手术、放射治疗和化疗。金属抗肿瘤药物是化疗药物的重要成员,但它们仍存在一些缺点,包括高发性的耐药性、较强的肾毒性和神经毒性以及对靶标DNA的低选择性等。
AIE(Aggregation-induced emission)即聚集诱导发光最初是2001年香港科技大学唐本忠院士首次提出。AIE类型的染料在稀溶液或均匀地分散在溶液中时,呈现出弱的荧光或几乎不发光,但当它在局部形成聚集态的时候,荧光被点亮。唐本忠院士提出了分子内旋转受限机制(Restriction of Intramolecular Motion,RIM),即化合物及其类似分子在分散状态下,激发的能量可以通过分子内苯环转动机械运动耗散无需辐射形式消耗,因此其在稀溶液状态下基本不发光。但当这些分子聚集起来,分子间相互作用增强,分子内旋转运动受限,激发态能量则通过辐射形式耗散,产生了发光现象。
能量代谢是肿瘤细胞抵抗凋亡、发生组织侵袭和转移等的关键环节,因此是抗癌药物潜在的作用靶标。通过调控肿瘤能量代谢可以改变肿瘤生长的微环境,切断能量供应途径,抑制肿瘤细胞增殖并促进其凋亡。线粒体是细胞内微小的细胞器,广泛参与信号传导、能量代谢、自噬和凋亡等细胞过程,通过产生ATP为细胞运转提供几乎全部所需的能量,对维持生物体正常生理功能至关重要。肿瘤细胞的线粒体在结构和功能上与正常细胞不同,因此,线粒体靶向化合物可能为肿瘤治疗提供一种诱导细胞死亡的有效手段。
发明内容
发明目的:本发明的目的之一是提供一种通过靶向肿瘤细胞线粒体,从而具有良好耐药性同时还能实现化疗和细胞成像联合作用的具有聚集诱导发光性质的线粒体靶向化合物;本发明另一目的是提供上述具有聚集诱导发光性质的线粒体靶向化合物的合成方法。
技术方案:本发明所述的具有聚集诱导发光性质的线粒体靶向化合物,所述化合物的结构式为:
上述具有聚集诱导发光性质的线粒体靶向化合物的合成方法,所述方法通过将具有三苯胺母体结构的有机化合物与三苯基膦反应,得到具有聚集诱导发光性质的线粒体靶向化合物;
其中,具有三苯胺母体结构的有机化合物的结构式为:
其中,所述具有三苯胺母体结构的有机化合物通过如下方法制备得到:在惰性氛围下,将溴吡啶苯酚和二甲基二氢丙啶溶解于有机溶剂中,于加热、搅拌条件下回流,反应完成后,通过萃取分离提取粗产物后,通过柱层析法提纯得到二甲基吡啶苯酚;
其中,溴吡啶苯酚的结构式为:
二甲基二氢丙啶的结构式为:
其中,所述具有三苯胺母体结构的有机化合物与三苯基膦的反应摩尔比为1:1.3~1.5。
其中,反应温度为60℃。
其中,所述溴吡啶苯酚和二甲基二氢丙啶的反应摩尔比为1:1.2~1.5。
其中,反应温度为60℃~80℃。
上述具有聚集诱导发光性质的线粒体靶向化合物在制备抗肿瘤药物中的应用。其中,肿瘤细胞为肺癌A549细胞。
上述具有聚集诱导发光性质的线粒体靶向化合物在制备抗顺铂耐受肿瘤细胞药物方面的应用。其中,顺铂耐受肿瘤细胞为肺癌A549R细胞。
其中,二甲基吡啶苯酚的化学反应式为:
其中,本发明化合物的化学反应式为:
有益效果:相比于现有技术,本发明具有的显著效果为:本发明通过将具有聚集诱导发光性质的荧光小分子化合物与线粒体靶向基团结合,所得到的化合物具有高抗癌活性(AIE分子本身无细胞器靶向,不具备抗癌活性,PPh3本身无抗癌活性,将两者结合在一起后得到具有线粒体靶向性的AIE分子聚集在线粒体后达到抗癌效果)和低毒副作用;首先,本发明化合物具有良好的细胞摄取能力(化合物能够快速在细胞内成像,因此认为化合物具有良好的细胞摄取能力)以及良好的靶向肿瘤细胞内线粒体的能力;其次,本发明化合物通过抑制人源肿瘤细胞增殖、诱导癌细胞内活性氧产生、降低线粒体膜电位、改变线粒体形态、损伤线粒体功能以及诱导肿瘤细胞(线粒体)自噬,实现良好的抗肿瘤活性;再次,由于本发明化合物的作用机制与金属抗肿瘤药物的作用机制不同,因此使用本发明化合物在抗肿瘤过程中不会产生耐药性的问题,从而保证持续性良好的抗癌效果;最后,本发明化合物能够有效抑制已对金属抗肿瘤药物(顺铂)产生耐药性的肿瘤细胞的增殖,从而解决现有化疗药物无法有效克服肿瘤细胞顺铂耐药性的问题。
附图说明
图1是实施例1化合物DP-PPh3的亚细胞器定位图;
图2是实施例1化合物DP-PPh3诱导A549R细胞产生活性氧的示意图;
图3是实施例1化合物DP-PPh3诱导A549R细胞线粒体膜电位下降的示意图;
图4是实施例1化合物DP-PPh3对线粒体形态及功能影响的示意图;图a为对照组和DP-PPh3药物组线粒体嵴结构变化对比;图b为A549R细胞经DP-OH药物组和DP-PPh3药物组处理24小时后氧气消耗速率(OCR)测试图;图c为线粒体功能相关的关键能量参数,基础呼吸、ATP合成、最大呼吸和备用呼吸的OCR值的定量柱状图;
图5是实施例1化合物DP-PPh3诱导A549R细胞自噬的示意图;
图6是实施例1化合物DP-PPh3诱导A549和A549R细胞中顺铂耐药相关蛋白含量变化的示意图;
图7是实施例1化合物DP-PPh3诱导3D肿瘤细胞球大量死亡的示意图。
具体实施方式
以下结合具体实施例对本发明的技术方案做进一步说明。
实施例1
本发明具有聚集诱导发光性质的线粒体靶向化合物(DP-PPh3)采用如下方法合成制得:
(1)在氩气保护氛围下,将Pd(OAc)2(23mg,0.1mmol)、t-BuONa(0.58g,6.0mmol)、(t-Bu)3PHBF4(88mg,0.3mmol)、溴吡啶苯酚(1.25g,5.0mmol)、二甲基二氢丙啶(DMAC,1.25g,6.0mmol)和甲苯(25mL)加入到100mL圆底瓶中(上述反应体系中,Pd(OAc)2、t-BuONa和(t-Bu)3PHBF4为催化剂);加热至60℃,回流搅拌48h,反应完成后冷却至室温,将反应物倒入水中,用50mL×3的二氯甲烷萃取,在无水Na2SO4上干燥;去除溶剂后,粗产物经硅胶柱层析,以DCM/乙酸乙酯(20:1v/v)为洗脱液,得到黄色固体二甲基吡啶苯酚(DP-OH),收集固体,真空干燥、称重(1.00g,得率为64%)。二甲基吡啶苯酚的结构式为:
通过核磁、质谱表征,1HNMR(400MHz,Methanol-d4)δ(ppm):8.89-8.83(m,2H),8.45-8.39(m,2H),7.59(t,J=2.0Hz,1H),7.51(dd,J=7.6,1.7Hz,2H),7.44(t,J=1.7Hz,1H),7.04(t,J=2.0Hz,1H),7.02-6.91(m,4H),6.36(dd,J=8.0,1.4Hz,2H),1.69(s,6H).ESI-MS(inCH3OH):理论值:m/z 378.17,实验值:378.48;
(2)在氩气保护氛围下,将二甲基吡啶苯酚(DP-OH)(37.8mg,0.1mmol)、三苯基膦TPP(62.2mg,0.13mmol)、K2CO3(41.5mg,0.3mmol)和丙酮(15mL)加入到50mL圆底烧瓶中;混合物料加热至60℃,回流搅拌12h;冷却至室温后,离心得到上清液,以二氯甲烷和无水乙醚为原料,经重结晶得到淡黄色固体DP-PPh3(61.3mg,产率79%);
DP-PPh3的结构式为:
通过核磁、质谱表征,1HNMR(400MHz,Chloroform-d)δ(ppm):8.71-8.66(m,2H),7.94-7.85(m,7H),7.75(dtd,J=8.4,6.9,1.7Hz,4H),7.67(td,J=7.5,3.4Hz,5H),7.62-7.57(m,2H),7.49(dd,J=7.4,1.9Hz,2H),7.36(t,J=1.9Hz,1H),7.23(d,J=1.7Hz,1H),6.98(dtd,J=15.5,7.3,1.7Hz,4H),6.35(dd,J=7.8,1.7Hz,2H),4.26(t,J=5.7Hz,2H),4.03(s,2H),2.37-2.25(m,2H),1.91(q,J=7.7Hz,2H),1.73(s,6H)。ESI-MS(inCH3OH):理论值:m/z 775.77,实验值:695.4。
将实施例1制得的化合物DP-PPh3与未进行线粒体靶向基团修饰的化合物DP-OH进行以下实验:
实施例1制得的化合物DP-PPh3对人源肺癌细胞A549及其顺铂耐药株A549R的抗肿瘤活性的应用:
利用MTT比色法来分析化合物DP-PPh3、化合物DP-OH和顺铂cis-Pt对人源肺癌细胞A549及其顺铂耐药株A549R的抗增殖效果。在活细胞中,线粒体内的琥珀酸脱氢酶可将MTT还原,生成一种蓝紫色产物:甲臜(可溶于DMSO),且该产物在570nm处有吸收峰,故可用A570 nm来分析细胞增殖情况。具体实验步骤为:(1)先复苏一管肿瘤细胞,用新鲜培养液(其中,A549细胞使用DMEM培养基+10%胎牛血清+1%盘尼西林和链霉素;A549R细胞使用RPMI-1640培养基+10%胎牛血清+1%盘尼西林和链霉素)培养,传代2次后使用;(2)待细胞到达对数生长期时,以5000个/孔的细胞密度接种至96孔板中(每孔200μL培养液),随即置于恒温箱(37℃,5%CO2)中培养;(3)待细胞贴壁后,每孔分别加入100μL含不同浓度梯度的化合物DP-PPh3、不同浓度梯度的化合物DP-OH及不同浓度梯度的顺铂cis-Pt的新鲜培养液,随即置于恒温箱中继续孵育;(4)孵育48小时后,在每孔中加入20μL MTT(5mg/mL),于37℃温箱中继续孵育4小时后,吸去上清液,每孔加入150μL二甲基亚砜(DMSO),使用酶联免疫检测仪检测A570nm,计算细胞增殖抑制率,求出IC50值(抑制率等于50%时对应的药物浓度)。化合物DP-PPh3、化合物DP-OH及顺铂cis-Pt的MTT测试结果如表1所示。
表1为化合物DP-PPh3、化合物DP-OH及顺铂cis-Pt的IC50值(μM)
aRF=resistance factor=IC50,A549R/IC50,A549.
结果表明:化合物DP-PPh3对A549和A549R的增殖抑制活性高于化合物DP-OH和cis-Pt,同时具有较低的RF值(RF,Resistance Factor,equals IC50(A549R)/IC50(A549)),表明经线粒体靶向基团三苯基膦修饰后的化合物DP-PPh3不仅对肺癌细胞A549具有好的抑制效果,对顺铂耐受的肺癌细胞A549R也具有好的抑制效果。
实施例1化合物DP-PPh3对细胞内亚细胞器(线粒体)定位的应用:
使用商业探针MitoRed、LysoRed检测化合物DP-PPh3在A54R9细胞中线粒体、溶酶体中的分布情况。往12孔板中的盖玻片上接种5×104个处于对数生长期的A549R细胞,置于培养箱中贴壁生长过夜,加入预先配制好的含有50μM化合物DP-PPh3的细胞培养液,继续孵育0.5小时。吸出培养液,用PBS洗涤三次。亚细胞器染色是在测试前分别加入含有一定浓度商业探针的培养液并孵育一定时间完成的,孵育时长为30min,随后在共聚焦显微镜下观察拍摄。化合物DP-PPh3:λex=405nm,λem=530±20nm;商业探针的激发和发射波长范围一致:λex=561nm,λem=600±20nm。共定位系数分析用ImageJ软件分析。
实施例1制得的DP-PPh3与线粒体探针和溶酶体探针共孵育后的共聚焦图如图1所示。结果表明:DP-PPh3与线粒体探针的共定位系数为0.84,而与溶酶体探针的共定位系数为0.11,表明DP-PPh3能够有效定位在A549R细胞内的线粒体中。
实施例1制得的DP-PPh3对诱发细胞内活性氧生成的应用:
方法1:共聚焦显微镜检测癌细胞内的ROS。A549R细胞接种于35mm的Corning激光共聚焦培养皿中,当细胞密度生长至70%时,分别加入化合物DP-PPh3(2μM)、化合物DP-OH(2μM)处理24h后,然后细胞用含10μMDCF-DA的无血清培养基于37℃下避光染色30min,随后立刻用共聚焦显微镜观察,激发波长为488nm,发射波长为530±20nm。
方法2:流式细胞仪检测癌细胞内的ROS。A549R细胞分别经化合物DP-PPh3(2μM)、化合物DP-OH(2μM)处理24h后,用含10μMDCF-DA的无血清培养基于37℃下避光染色30min,离心弃上清,用无血清的培养基洗涤三次,除去未进入细胞内的DCF-DA;收集细胞后半小时内用流式细胞仪测定绿色荧光强度;激发波长为488nm,发射波长为530±20nm。绿光的平均荧光强度用FlowJo 7.6(Tree Star,OR,USA)软件分析。
化合物DP-PPh3对诱发细胞内活性氧生成的结果如图2所示。结果表明:与对照组(化合物DP-OH)相比,经过化合物DP-PPh3处理后可以有效诱发细胞内活性氧含量升高,A549R细胞加入2μM化合物DP-PPh3后,细胞内活性氧物质增加10倍左右,同时共聚焦图中绿色荧光明显增强。而DP-OH处理组的细胞内活性氧含量无明显变化,这说明化合物DP-PPh3进入癌细胞后会诱导癌细胞产生活性氧物质(ROS),从而导致癌细胞发生死亡。
实施例1制得的DP-PPh3对诱发细胞内线粒体膜电位变化的应用:
方法1:共聚焦显微镜检测肿瘤细胞内线粒体膜电位的变化。A549R细胞接种于35mm的Corning激光共聚焦培养皿中,当细胞密度生长至70%时,分别加入化合物DP-PPh3(2μM)、化合物DP-OH(2μM)处理24h后,然后细胞用预先配置好的JC-1工作液在37℃下避光染色30min,随后立刻用共聚焦显微镜观察。
方法2:流式细胞仪检测肿瘤细胞内线粒体膜电位的变化。将含有化合物DP-PPh3(2μM)、化合物DP-OH(2μM)的细胞培养液分别加至接种有形态良好且长势正常的A549R细胞的6孔板中,药物处理24h后,收取细胞,PBS洗涤,随后加入配制好的JC-1工作液染色25-30min;用buffer洗涤细胞并重悬,立即使用BD FACSverse流式细胞仪检测待测样品,并用FlowJo 7.6软件处理所得结果并进行分析。检测荧光通道为λex=488nm,λem=530±30nm;λex=488nm,λem=590±30nm。
化合物DP-PPh3对诱发细胞内线粒体膜电位变化的结果如图3所示。结果表明:与对照组(化合物DP-OH)相比,经过化合物DP-PPh3处理后可以观察到细胞内的红色荧光减弱,绿色荧光明显增强,而DP-OH处理组的线粒体膜电位无明显变化,表明化合物DP-PPh3有效地诱发线粒体膜电位下降,同时在流式细胞仪的结果也表明相同的结论。
实施例1制得的化合物DP-PPh3对线粒体形态及功能的影响:
通过透射电子显微镜(TEM)对化合物DP-PPh3处理后A549R细胞的线粒体形态进行观察。往已接种有A549R细胞的100mm培养皿中,加入预先配制好的含有2μM化合物DP-PPh3的细胞培养液进行药物处理,待共孵育24h后,收取细胞,用PBS洗涤,2.5%戊二醛进行固定,用酒精进行梯度脱水,用树脂进行包埋,超薄切片,使用乙酸铀酰和柠檬酸铅染色,并在铜网上制备待观测样品,使用Hitachi H-7650透射显微镜对线粒体形态进行观察。
采用XF24细胞线粒体氧压力测试试剂盒对化合物DP-PPh3(2μM)、化合物DP-OH(2μM)处理后的A549R细胞的氧气消耗速率(OCR)进行测试。将处于对数生长期的A549R细胞,以4×104/孔接种于XF24孔细胞培养板中,待其贴壁生长后,往每孔加入250μL预先配制好的含有化合物DP-PPh3、化合物DP-OH的细胞培养液进行药物处理24h;配制含有葡萄糖(25mM)和丙酮酸(2mM)的XF检测液并将其pH调至7.4;待药物孵育结束后,用新鲜配制的XF检测液洗涤细胞3次,然后向每孔加入450μL含有相应药物浓度的XF检测液并置于37℃、无CO2的细胞培养箱中继续孵育1h;在预先水化好的探针板上所有A孔中加入75μLATP合成酶抑制剂寡霉素(Oligomycin,1μM)、所有B孔中加入75μL解偶联剂(FCCP,1μM)、所有C孔中加入75μL呼吸链抑制剂2-脱氧-D-葡萄糖抗霉素A+鱼藤酮(AntimycinA+Rotenone,1μM);使用SeahorseXFe24细胞生物分析仪进行OCR测试。测试结束后,使用BCA蛋白浓度测定试剂盒对细胞培养板中的每孔细胞进行蛋白含量测定,计算线粒体呼吸功能的参数。
化合物DP-PPh3对线粒体形态及功能的影响的TEM及OCR实验结果如图4所示。结果表明:与未加药处理的Control组完整的线粒体嵴结构相比,化合物DP-PPh3处理后的A549R细胞线粒体肿胀变圆,嵴结构消失,表明化合物DP-PPh3损伤破坏线粒体结构形态。OCR是氧化磷酸化(OXPHOS)的一个重要的指标,与线粒体功能相关的关键能量参数,包括分别用于基础呼吸、ATP合成、最大呼吸和备用呼吸的OCR值。在化合物DP-PPh3作用下,A549R细胞上述关键能量参数均显著降低,而DP-OH处理后上述关键能量参数无明显变化,表明化合物DP-PPh3通过抑制线粒体的基础呼吸和ATP产生等来降低A549R细胞的线粒体呼吸功能,从而抑制细胞的OXPHOS过程。
实施例1制得的DP-PPh3对诱导A549R细胞自噬的应用:
利用蛋白免疫印迹(WB)检测自噬蛋白含量变化。往已贴壁生长A549R细胞的100mL培养皿中加入预先配制好的含有化合物DP-PPh3(2μM,4μM)、化合物DP-OH(2μM)的细胞培养液,待药物处理48后,离心收集细胞,PBS洗涤以去除残留培养液中的血清,加入含有PMSF的碧云天RIPA强裂解液进行20min的全细胞裂解,在这整个过程中保证4℃低温环境以确保蛋白不变性。结束后在低温条件下以13400rpm转速离心20min,吸取离心所得上清液即是实验所需的细胞全蛋白样品;使用BCA蛋白含量检测试剂盒确定上述蛋白样品中的蛋白浓度;利用SDS-PAGE凝胶电泳对样品内不同蛋白表达含量进行检测。制好胶之后,向每个孔中加入相同体积的蛋白样品进行凝胶电泳实验,适当分离后立即停止电泳;使用湿法将目的蛋白转至PVDF膜上,结束后,将膜置于含有5%脱脂奶粉中封闭2h。根据抗体的使用说明按照相应的比例用脱脂奶粉对一抗进行稀释,将封闭好的膜置于一抗孵育液中室温孵育一段时间使其与目的蛋白特异性地结合。结束后,用PBST洗涤(5×6min/次)。将洗干净的膜置于预先配制好的二抗孵育液中孵育一段时间使其与一抗结合,同样用PBST进行洗涤。配制等体积的ECL显影液,将其覆盖于PVDF膜上,处理2min后利用化学发光成像系统进行拍摄。
化合物DP-PPh3对A549R细胞诱导细胞自噬相关蛋白表达的实验结果如图5所示。结果表明:与未加药处理的对照组或DP-OH药物组相比,细胞经过DP-PPh3处理后,线粒体自噬相关蛋白PINK1和Parkin表达明显上调,同时自噬标志性蛋白LC3-II蛋白含量呈浓度依赖性增加,说明化合物DP-PPh3诱导A549R细胞的死亡方式为自噬。
实施例1制得的DP-PPh3在克服肿瘤细胞顺铂耐药机制的应用:
通过蛋白免疫印迹(WB)检测化合物DP-PPh3处理过的A549和A549R细胞中与克服肿瘤顺铂耐药相关蛋白含量变化。本应用中所使用的一抗包括泵入相关蛋白CTR1和泵出相关蛋白MRP2。
化合物DP-PPh3对泵入相关蛋白CTR1和泵出相关蛋白MRP2蛋白表达变化的实验结果如图6所示。结果表明:在A549R细胞中,与对照组相比,经过DP-PPh3处理后,细胞样品中MRP2蛋白表达明显下调,CTR1蛋白表达明显上调。在A549细胞中,观察到完全相反的结果,即与对照组相比,DP-PPh3处理后的细胞样品中MRP2蛋白表达略微上调,CTR1蛋白表达无明显变化。说明DP-PPh3是通过调控泵入相关蛋白CTR1和泵出相关蛋白MRP2的表达克服肿瘤细胞顺铂耐药性。
实施例1制得的DP-PPh3的3D细胞球抗肿瘤活性的应用:
将处于对数生长期的A549R细胞,以2.5×103/孔/100μL接种于3D细胞球培养的96孔板中,置于37℃培养箱中培养5天后至细胞球的大小直径达到400μm。往每孔加入100μL预先配制好的含有化合物DP-PPh3(20μM)、化合物DP-OH(20μM)的细胞培养液进行药物孵育处理。孵育9天后,利用Calcein AM/PI染色试剂盒对细胞球的活死状态进行标记,其中Calcein AM标记活细胞,PI标记死细胞。激发和发射波长分别为:Calcein AM:λex=488nmandλem range 500-550nm,PI:λex=561nm andλem range 570-620nm,随后利用激光共聚焦显微镜进行Z-stack扫描拍摄。
化合物DP-PPh3对3D细胞球抗肿瘤活性实验结果如图7所示。结果表明:与对照组或DP-OH药物组相比,细胞球经过DP-PPh3处理后,绿色标记的活细胞数量明显减少,红色标记的死细胞数量大量增加,表明经线粒体靶向基团三苯基膦修饰后的化合物DP-PPh3对顺铂耐受的肺癌3D细胞球具有较好的抑制效果,诱导3D细胞球产生大量死亡。
Claims (9)
1.一种具有聚集诱导发光性质的线粒体靶向化合物,其特征在于,所述化合物的结构式为:
2.权利要求1所述的具有聚集诱导发光性质的线粒体靶向化合物的合成方法,其特征在于:所述方法通过将具有三苯胺母体结构的有机化合物与三苯基膦反应,得到具有聚集诱导发光性质的线粒体靶向化合物;
其中,具有三苯胺母体结构的有机化合物的结构式为:
3.根据权利要求2所述的具有聚集诱导发光性质的线粒体靶向化合物的合成方法,其特征在于,所述具有三苯胺母体结构的有机化合物通过如下方法制备得到:在惰性氛围下,将溴吡啶苯酚和二甲基二氢丙啶溶解于有机溶剂中,于加热、搅拌条件下回流,反应完成后,通过萃取分离提取粗产物后,通过柱层析法提纯得到具有三苯胺母体结构的有机化合物;
其中,溴吡啶苯酚的结构式为:
二甲基二氢丙啶的结构式为:
4.根据权利要求2所述的具有聚集诱导发光性质的线粒体靶向化合物的合成方法,其特征在于:所述具有三苯胺母体结构的有机化合物与三苯基膦的反应摩尔比为1:1.3~1.5。
5.根据权利要求2所述的具有聚集诱导发光性质的线粒体靶向化合物的合成方法,其特征在于:反应温度为60~65℃。
6.根据权利要求3所述的具有聚集诱导发光性质的线粒体靶向化合物的合成方法,其特征在于:所述溴吡啶苯酚和二甲基二氢丙啶的反应摩尔比为1:1.2~1.5。
7.根据权利要求3所述的具有聚集诱导发光性质的线粒体靶向化合物的合成方法,其特征在于:反应温度为60℃~80℃。
8.权利要求1所述的具有聚集诱导发光性质的线粒体靶向化合物在制备抗肿瘤药物中的应用。
9.权利要求1所述的具有聚集诱导发光性质的线粒体靶向化合物在制备抗顺铂耐受肿瘤细胞药物方面的应用。
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