CN110404080A - 一种线粒体靶向的磷脂及其应用 - Google Patents
一种线粒体靶向的磷脂及其应用 Download PDFInfo
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- CN110404080A CN110404080A CN201910613285.9A CN201910613285A CN110404080A CN 110404080 A CN110404080 A CN 110404080A CN 201910613285 A CN201910613285 A CN 201910613285A CN 110404080 A CN110404080 A CN 110404080A
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- Prior art keywords
- phosphatide
- arginine
- lysine
- polypeptide
- mitochondrially targeted
- Prior art date
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Links
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- 238000005303 weighing Methods 0.000 description 1
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Abstract
本发明设计构建了由一类具有病灶部位靶向蓄积、降低非特异性毒性、病灶组织高渗透性及线粒体靶向的磷脂,磷脂的亲水头部通过酰胺键或酯键连接(3‑丙羧基)三苯基磷、精氨酸、赖氨酸或由精氨酸和/或赖氨酸组成的多肽。本发明所述的磷脂可应用于药物递送载体,能够使得药物在组织中高效富集、高度渗透及靶向细胞线粒体。
Description
技术领域
本发明属于药物递送系统领域,涉及一种线粒体靶向的磷脂,特别涉及一种 pH敏感且线粒体靶向磷脂及其应用。
背景技术
线粒体在真核细胞的生理和病理发育过程中发挥着重要作用,其功能障碍与多种疾病的发生、发展过程密切相关。线粒体靶向治疗的关键是有效地将生物活性分子传递到线粒体内部。
在生理条件下线粒体进行新陈代谢所需要的蛋白质也需要在细胞质内合成然后再从细胞质运送到线粒体内。目前,线粒体靶向信号肽(MTSs)、蛋白转导域(PTD)和离域亲脂阳离子(如三苯基膦、TPP)通常作为线粒体靶向基团将生物活性物质运输到线粒体中。然而,非特异性毒性和肿瘤组织分布不均匀仍然是线粒体靶向药物临床应用所面临的主要挑战。在生理条件下,线粒体进行新陈代谢所需要的蛋白质也需要在细胞质内合成,然后再从细胞质运送到线粒体内。这为寻找靶向线粒体的药物载体提供了新的思路。从线粒体蛋白中分离出来的多肽序列叫做线粒体引导序列。这些多肽序列一般由10-80个阳离子、疏水性或者一些特殊氨基酸构成。(KLAKLAK)2是从细胞促凋亡蛋白里分离出来的一个多肽序列,可以有效应用于线粒体定位。Flierl AJackson等通过将小分子量的外源 DNA连接到(KLAKLAK)2,成功地将外源DNA导入了线粒体(Targeted delivery of DNA to themitochondrial compartment via import sequence conjugated peptide nucleicacid.[J].Mol Ther,2003,7:550-557)。然而阳离子物质的非特异性毒性,极差的病灶部位渗透性及复杂的多肽合成过程,严重了制约了该类物质的临床转化及应用。上述问题代表了目前整个线粒体递送系统存在的问题。
发明内容
本发明针对现有技术不足,在磷脂的结构基础上,设计并构建了由一类具有病灶部位靶向蓄积、降低非特异性毒性、病灶组织高渗透性及线粒体靶向的磷脂,将其应用于药物递送载体,能够使得药物在组织中高效富集、高度渗透及靶向细胞线粒体。
本发明具体技术方案如下:
一种线粒体靶向的磷脂,磷脂的亲水头部通过酰胺键或酯键连接(3-丙羧基)三苯基磷、精氨酸、赖氨酸或由精氨酸和/或赖氨酸组成的多肽。
所述磷脂是指含有磷酸的脂类,分为甘油磷脂与鞘磷脂两大类,分别由甘油和鞘氨醇构成。包括磷脂酰胆碱(卵磷脂)、磷脂酰乙醇胺(脑磷脂)、磷脂酰丝氨酸、磷脂酰甘油、二磷脂酰甘油(心磷脂)及磷脂酰肌醇、神经鞘磷脂等。
磷脂亲水头部的羟基或者氨基可以与(3-丙羧基)三苯基磷、精氨酸、赖氨酸或由精氨酸和/或赖氨酸组成的多肽的羧基反应,形成酰胺键或者酯键。
优选的,所述多肽为精氨酸和/或赖氨酸通过酰胺键组成的树形多肽:
(n代表精氨酸、赖氨酸,以酰胺键形成树形网络)。
优选的,组成树形多肽的氨基酸数量为2-10,更优选2-5,选用低代数的磷脂可以使得材料合成简单易控,满足工业化生产的高重复性及高产量。同时较低代数的树形分子多肽构建的磷脂,利用化学自主装的过程依然可以展现高代数的树形分子的优点,有可以降低高代数自身的毒性,提高使用的安全性。
本发明所述的磷脂,一个具体的方案,以亲水部分为磷脂酰乙醇胺为例,具有如下结构:
其中R1和R2相同或者不同,代表-CnH2n+1或-CnH2n,n=1~18的整数,R3为精氨酸、赖氨酸或由精氨酸和/或赖氨酸组成的多肽的C端形成酰胺键与母核结构连接,优选代表
本发明另一目的在于提供一种pH敏感且线粒体靶向磷脂,在上述磷脂的基础上,所述精氨酸、赖氨酸或由精氨酸和/或赖氨酸组成的多肽的游离氨基上还连接有 R4,代表a=0,1,2,b=0,1,2,优选的,R4代表
本发明优选的磷脂结构如下:
化合物1 G1R:
化合物2 G2R:
化合物3 G2K:
化合物4 DTPP:
化合物5 G1R-R2:
化合物6 G2R-R2:
化合物7 G2K-R2:
本发明所述的线粒体靶向的磷脂可以采用酰胺缩合的方法制备而成。
本发明另一目的在于提供本发明所述的磷脂在制备线粒体靶向药物递送系统中的应用。本发明所述的磷脂可以作为脂质材料/乳化剂用于制备药物载体,如脂质体、微乳、微球、微囊等,可以承载降低或提高线粒体氧化应激能力的药物,可用于治疗肿瘤、动脉粥样硬化,急性肝所伤,抗皮肤光衰老等。
本发明一个具体的实施方式,以光敏剂吲哚菁绿为药物,设计构建了修饰有酸可裂解的2,3-二甲基顺丁烯二酸的二代树状精氨酸磷脂G2R-DA,将G2R-DA、大豆磷脂酰胆碱(SPC)、胆固醇、聚乙二醇-二硬脂酰磷脂酰乙醇胺(DSPE-PEG2000) 和光敏剂吲哚菁绿(ICG)联合组装得到脂质体(L-G2R-DA),用于线粒体靶向肿瘤治疗,系统评价了吲哚菁绿诱导光动力(PDT)和光热肿瘤治疗(PTT)的体内外生物学效应。结果显示,使用本发明所述磷脂制备的脂质体,不仅可以控制药物在肿瘤组织和正常组织中的分布量,而且能够使得药物在肿瘤组织中高效富集、高度渗透及靶向肿瘤细胞线粒体。
本发明优点:
(1)本发明使用精氨酸、赖氨酸或由精氨酸和/或赖氨酸组成的肽类树形分子模
(2)同时,利用2-3-二甲基马来酸酐修饰氨基酸的氨基或胍基,构建对生物刺激响应的可控/随需暴露系统,以减少非特异性毒性。
(3)此外,精氨酸的分子可以大大提高细胞摄入量和肿瘤组织的均匀分布,从而获得更好的治疗效果。
附图说明
图1是实施例1-4所述的线粒体靶向的磷脂制备的脂质体的线粒体靶向性能测定。
图2是实施例1-4所述的线粒体靶向的磷脂制备的脂质体的线粒体靶向性能流式定量测定结果。
图3是实施例5所述的pH敏感性、线粒体靶向的磷脂制备的脂质体的细胞摄取图。
图4是实施例5所述的pH敏感性、线粒体靶向的磷脂制备的脂质体的3D细胞渗透图。
图5是实施例5所述的pH敏感性、线粒体靶向的磷脂制备的脂质体的动物体内分布图。
图6是实施例5所述的pH敏感性、线粒体靶向的磷脂制备的脂质体的抗肿瘤效果图。
具体实施方式
以下通过实施例说明本发明的具体步骤,但不受实施例限制。
在本发明中所使用的术语,除非另有说明,一般具有本领域普通技术人员通常理解的含义。
下面结合具体实例并参照数据进一步详细描述本发明。应理解,这些实施例只是为了举例说明本发明,而非以任何方式限制本发明的范围。
在以下实施例中,未详细描述的各种过程和方法是本领域中公知的常规方法。
实施例1
本实施例中,所述线粒体靶向的磷脂为二硬脂酰磷脂酰乙醇胺-精氨酸 (G1R),其结构式为:
其制备方法如下:
取DSPE 0.75g(0.75mmol),在氮气保护下溶于15mL无水三氯甲烷。另取1mL DIPEA(6.00mmol)在0℃下搅拌加入DSPE溶液中。然后将0.79g Boc-Arg(Pbf)-OH(0.79mmol)、0.2g HOBT(0.2mmol)和0.57g HBTU(1.5mmol) 共同溶于2.5mL的DMF中,添加到包含DSPE反应烧瓶,然后在室温下继续反应24小时。待反应结束后,将反应产物依次使用饱和碳酸氢钠、硫酸氢钠和氯化钠溶液反复洗涤数次后,再将产物用硫酸镁干燥2小时。最后将溶剂除去后, 采用硅胶柱层析法纯化,洗脱剂为二氯甲烷/甲醇(12/1,v/v),收集产物。而后将其进行真空干燥,使用10mL DCM/TFA的混合溶液(1:1,v/v)处理4小时以除去叔丁基基团。产物浓缩,并用无水乙醚处理得到G1R,产率为90%。
实施例2
本实施例中,所述线粒体靶向的磷脂为二硬脂酰磷脂酰乙醇胺-二代精氨酸(G2R),其结构式为:
其制备方法如下:
称取H-Lys-OMe·2HCl 2.00g(8.6mmol),Boc-Arg(Pbf)-OH 13.56g(13.56mmol),HBTU 9.78g(25.8mmol),HOBT 3.36g(3.36mmol),在氮气保护下共同溶解于50mL无水DMF中,然后在冰水浴条件下将11.4mL的DIPEA(68.8 mmol)加入上述混合液中,在室温下搅拌反应48h。然后,将产物分别用饱和 NaHCO3、NaHSO4和NaCl溶液洗涤数次。接着将产物与MgSO4混合2h除去水分。除去溶剂后,使用硅胶柱层析法对产物进行纯化,洗脱剂为二氯甲烷:甲醇(12:1,v/v),收集产物。称取产物8.00g(6.8mmol)于100mL含有1mol/L NaOH 的甲醇溶液中,与氢氧化钠反应4h,从而发生脱羧反应。待甲醇完全挥发后,将产物溶于水中,并调节pH值近中性。最后使用二氯甲烷萃取出产物并同样使用硫酸镁干燥2小时。最后称取DSPE 1.5g(2.00mmol)于反应瓶中,在氮气保护下,溶于30mL无水三氯甲烷中。量取DIPEA2.00mL(12.00mmol)在0℃搅拌下加入上述DSPE溶液中。然后称取上一步产物(3mmol)与0.4g(3mmol) HOBT和1.14g(4mmol)HBTU于反应瓶中,并用5mL无水DMF溶解。氮气保护室温下继续反应24小时,待反应结束后,将反应产物依次使用饱和碳酸氢钠、硫酸氢钠和氯化钠溶液反复洗涤数次后,再将产物用硫酸镁干燥2小时。最后将溶剂除去后,采用硅胶柱层析法纯化,洗脱剂为二氯甲烷/甲醇(12/1,v/v),收集产物并进行真空干燥后,使用10mL DCM/TFA混合溶液(1:1,v/v)处理4 小时以出去叔丁基基团。然后将产物浓缩,并用无水乙醚处理得到G2R。
实施例3
本实施例中,所述线粒体靶向的磷脂为二硬脂酰磷脂酰乙醇胺-二代赖氨酸(G2K),其结构式为:
其制备方法如下:
将H-Lys-OMe.2HCl(1.00g,4.3mmol)、Boc-L-lys(Boc)-OH(4.46g,12.9 mmol)、HBTU(4.89g,12.9mmol)、HOBT(1.68g,12.9mmol)溶于无水DMF(25 mL)中。冰水浴中加入DIPEA (5.7mL,34.4mmol)并在室温下搅拌48小时,然后用饱和NaHCO3、NaHSO4、NaCl溶液对混合溶液进行多次洗涤。用MgSO4干燥2h,溶剂去除后,用硅胶柱层析(DCM/MeOH,12/1,v/v)纯化得到产物。取其(4.00g,3.4mmol)置于50mLNaOH的MeOH(1mol/L)溶液中处理4h,暴露羧基。去除MeOH后,将混合物溶解在H2O中,调节至中性pH值。经DCM 萃取,MgSO4干燥2h。取产物(1.81g,1.5mmol)、DSPE(0.75g,1.00mmol)、 HOBT(0.27g,2mmol)、HBTU(0.57g,2mmol)分别溶于无水三氯甲烷(25mL) 和无水DMF(3mL)中,氮气保护。将DIPEA(2.00mL,14.00mmol)加入上述混合溶液中,在0℃搅拌,并且氮气下室温继续反应48小时。用饱和的NaHCO3、 NaHSO4和NaCl溶液对混合物进行多次洗涤。用MgSO4干燥2h,去除溶剂后,用硅胶柱层析(DCM/MeOH,12/1,v/v)纯化得到产物,并将其在真空中干燥,溶于无水二氯甲烷DCM/TFA (1:1,10ml)中4h,以脱去叔丁基。将混合物浓缩,用无水乙醚对产物进行处理,得到G2K。
实施例4
本实施例中,所述线粒体靶向的磷脂为二硬脂酰磷脂酰乙醇胺-(3-丙羧基) 三苯基磷(DTPP),其结构式为:
其制备方法如下:
先将DSPE 0.1g(0.133mmol)在氮气保护下溶于15mL三氯甲烷中,然后在0℃搅拌下将0.25mL DIPEA(1.3mmol)加入至DSPE溶液中。另取(3-丙羧基)三苯基溴化磷(3-carboxypropyl triphenyl-phosphonium bromide,3-CTPP) 0.084g(0.19mmol)、HOBT0.025g(0.19mmol)和HBTU 0.071g(0.19mmol),溶解于2.5mL无水DMF(2.5毫升),然后将其加入含DSPE的反应瓶中。在氮气保护下,冰浴搅拌30min并于室温下继续反应24h。反应结束后,将混合物依次用饱和碳酸氢钠、硫酸氢钠和氯化钠溶液洗涤数次后,再用硫酸镁干燥2h。最后除去溶剂,采用硅胶柱层析法进行纯化,洗脱剂为二氯甲烷/甲醇(12/1,v/v),即可获得DTTP。
实施例5
本实施例中,所述pH敏感、线粒体靶向的磷脂为二硬脂酰磷脂酰乙醇胺- 二代精氨酸-2,3二甲基马来酸酐(G2R-DA),其结构式为:
其制备方法如下:
称取实施例2中的G2R 1g(0.84mmol),溶解在10mL无水二氯甲烷中。然后称取TEA1.77g(1.77mmol)、DMA 2.2g(2.2mmol)加入上述溶液中。在氮气保护下,将各反应物在冰浴下孵育30min,然后在室温下反应24小时。接着,再次向反应产物中加入DIPEA 1.77g(1.77mmol)和DMA 2.2g(2.2mmol),继续在室温下反应24小时。反应结束后,将产物逐滴加入100mL的冰无水乙醚中,待产物完全形成沉淀后,将沉淀溶解于甲醇中,并用透析袋(MWCO= 1000)在4℃下透析3天,透析介质为pH 7.4的去离子水,并每隔4小时更换一次透析介质。透析结束后,将产物进行冷冻干燥,得到G2R-DA。
不具有pH响应性的对照材料G2R-SA也被设计,其制备过程如下:称取实施例2中的G2R 1g(0.84mmol),溶解在10mL无水二氯甲烷中。然后称取TEA 3.54g(3.54mmol)、SA 4.4g(19.4mmol)加入上述溶液中。在氮气保护下,将各反应物在冰浴下孵育30min,然后在室温下反应24小时。接着,再次向反应产物中加TEA 3.54g(3.54mmol)、SA 4.4g(19.4mmol)加入上述溶液中,继续在室温下反应24小时。反应结束后,将产物逐滴加入100mL的冰无水乙醚中,待产物完全形成沉淀后,将沉淀溶解于甲醇中,并用透析袋(MWCO= 1000)在4℃下透析3天,透析介质为pH 7.4的去离子水,并每隔4小时更换一次透析介质。透析结束后,将产物进行冷冻干燥,得到G2R-SA。
将实施例1-4制备的材料分别与大豆磷脂、胆固醇以及Dil (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate)溶于25mL氯仿- 甲醇混合溶剂中(氯仿与甲醇的体积比为2:1),然后减压旋转蒸发除去溶剂,得到干燥的脂膜。然后加入去离子水,在37℃水浴中超声10min,继以探头超声5min,得到脂质体溶液。将4T1细胞以104个/孔培养接种于玻璃小皿中,孵育24h后,将载Dil的各脂质体(含Dil 1mM/孔)加入各玻璃小皿,并继续培养12h。在激光共聚焦扫描显微镜下观察4T1细胞中各种Dil负载脂质体的线粒体定位。用线粒体绿色荧光探针进行染色,并用激光共聚焦观察。线粒体经绿色荧光染料Mitotracker Green FM(特异性线粒体选择性染料)染色后呈绿色荧光,脂质体荷载Dil后呈红色荧光。黄色表示脂质体和线粒体的共定位情况。如图1 所示,亮黄色荧光表明实施例1~4所述的线粒体靶向磷脂制备的脂质体均具有较好的线粒体靶向性能,特别是G2R构建的脂质体L-G2R显示出最强的黄色荧光,说明其具有最佳的线粒体靶向效率。如图2流式定量数据也证明了上述结果,同时表明L-G2R的线粒体靶向性能是L-TPP的6.2倍,L-G1R的5.7倍以及L-G2K 的4.4倍,G2R能够令脂质体具有优良的线粒体靶向性能。
实施例7
将实施例5制备的材料分别与大豆磷脂、胆固醇以及Dil溶于25mL氯仿- 甲醇混合溶剂中(氯仿与甲醇的体积比为2:1),然后减压旋转蒸发除去溶剂,得到干燥的脂膜。然后加入去离子水,在37℃水浴中超声10min,继以探头超声5min,得到脂质体溶液。将4T1细胞以4×105/孔接种于6孔板,待细胞融合至80%后,用PBS洗涤,更换含新鲜培养基。以空白细胞为参比,Dil药物溶液为阴性对照,不同载Dil的脂质体为实验组,将各组脂质体加入各孔中(含 Dil 1mM/孔),分别在pH 7.4或pH 6.8的培养基中37℃孵育12h后,吸弃各孔内含Dil的培养基,用PBS缓冲液洗涤3次,胰酶消化,1000r/min离心10min 收集4T1细胞,最后加入0.5ml PBS重悬,流式细胞仪上机检测Dil荧光强度。如图3所示,实施例5所述的pH敏感、线粒体靶向磷脂制备的脂质体,其在pH 6.8的弱酸性培养条件下培养比pH 7.4下培养增强了约10倍的药物摄取量,具有弱酸环境促进细胞摄取的优良性能。而对照组L-PEG和L-G2K-SA未表现出 pH响应性能,pH的变化对摄取无影响。
实施例8
将实施例5制备的材料分别与大豆磷脂、胆固醇以及Dil溶于25mL氯仿- 甲醇混合溶剂中(氯仿与甲醇的体积比为2:1),然后减压旋转蒸发除去溶剂,得到干燥的脂膜。然后加入去离子水,在37℃水浴中超声10min,继以探头超声5min,得到脂质体溶液。取已构建好的4T1多细胞球体MCs约150μm悬浮液(含有约20个球状体)至10mL离心管中。分别将载Dil的各脂质体(含1mM Dil)加入各离心管中,并在37℃的CO2下孵育12h。在设定的各时间点,用PBS 洗涤MCs并转移至共焦培养皿中。使用激光共聚焦从上到下每隔15μm进行共聚焦成像观察。如图4所示,实施例5所述的pH敏感、线粒体靶向磷脂制备的脂质体能够渗透到3D肿瘤细胞球的内部,近肿瘤球中心的横切面显示出最强的荧光信号,说明其具有优良的渗透性能。而对照组游离药物、L-PEG和L-G2K-SA 均渗透性较差。
实施例9
将实施例5制备的材料分别与大豆磷脂、胆固醇以及ICG溶于25mL氯仿- 甲醇混合溶剂中(氯仿与甲醇的体积比为2:1),然后减压旋转蒸发除去溶剂,得到干燥的脂膜。然后加入去离子水,在37℃水浴中超声10min,继以探头超声5min,得到脂质体溶液。以7.5mg/kgICG的剂量向荷4T1肿瘤的小鼠静脉分别注射游离ICG和载ICG的脂质体。在静脉内注射4h,8h,12h,24h和48h 后用活体成像系统对小鼠进行荧光成像观察并拍照。如图5所示,实施例5所述的pH敏感、线粒体靶向磷脂制备的脂质体在荷瘤小鼠的肿瘤中显示出最强的荧光信号,具有优良的药物累积性能且具有长循环的特点。而对照组游离药物、 L-PEG和L-G2K-SA累积性能及长循环性能较差。
实施例10
将实施例5制备的材料分别与大豆磷脂、胆固醇以及ICG溶于25mL氯仿- 甲醇混合溶剂中(氯仿与甲醇的体积比为2:1),然后减压旋转蒸发除去溶剂,得到干燥的脂膜。然后加入去离子水,在37℃水浴中超声10min,继以探头超声5min,得到脂质体溶液。以7.5mg/kgICG的剂量向荷4T1肿瘤的小鼠静脉分别注射游离ICG和载ICG的脂质体。将荷4T1肿瘤的BALB/c小鼠随机分成8 组,每组小鼠5只。阴性对照组:PBS,实验组:ICG/L-PEG,ICG/L-G2R-SA 和ICG/L-G2R-DA。以7.5mg/kg ICG的剂量,分别在第0天、第2天通过尾静脉注射至荷瘤小鼠体内,每天注射2次。在静脉注射8h后用波长为808nm、光照强度为1.5W/cm2的激光持续照射或非持续照射小鼠5min。还对每隔两天对小鼠进行称重并对肿瘤组织直径进行测量。如图6所示,实施例5所述的pH敏感、线粒体靶向磷脂制备的脂质体,结合光热及光动治疗,可以令肿瘤完全消除,具有十分优良的抗肿瘤效果;且其体重无明显变化,证明该材料具有较低的系统毒性。而对照组游离药物、L-PEG和L-G2K-SA抗肿瘤效果较差。
Claims (10)
1.一种线粒体靶向的磷脂,其特征在于磷脂的亲水头部通过酰胺键或酯键连接(3-丙羧基)三苯基磷、精氨酸、赖氨酸或由精氨酸和/或赖氨酸组成的多肽。
2.如权利要求1所述的磷脂,其特征在于所述多肽为精氨酸和/或赖氨酸通过酰胺键组成的树形多肽。
3.如权利要求2所述的磷脂,其特征在于所述组成树形多肽的氨基酸数量为2-10。
4.如权利要求3所述的磷脂,其特征在于所述组成树形多肽的氨基酸数量为2-5。
5.如权利要求1所述的磷脂,其特征在于具有如下结构:
其中R1和R2相同或者不同,代表-CnH2n+1或-CnH2n,n=1~18的整数,R3为精氨酸、赖氨酸或由精氨酸和/或赖氨酸组成的多肽的C端形成酰胺键与母核结构连接。
6.如权利要求1所述的磷脂,其特征在于R3代表
7.如权利要求1-6任一项所述的磷脂,其特征在于所述精氨酸、赖氨酸或由精氨酸和/或赖氨酸组成的多肽的游离氨基上还连接有R4,代表a=0,1,2,b=0,1,2。
8.如权利要求7所述的磷脂,其特征在于所述R4代表
9.如权利要求1-8任一项所述的磷脂在制备线粒体靶向药物递送系统中的应用。
10.如权利要求9所述的应用,其特征在于所述药物降低或提高线粒体氧化应激能力的药物。
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1988000824A1 (en) * | 1986-07-28 | 1988-02-11 | Liposome Technology, Inc. | Liposomes with enhanced retention on mucosal tissue |
CN104193779A (zh) * | 2014-08-22 | 2014-12-10 | 四川大学 | 一种pH敏感磷脂药用材料及其制备方法与应用 |
CN105801668A (zh) * | 2016-04-01 | 2016-07-27 | 天津大学 | 一种寡聚精氨酸修饰的磷脂与其组装的纳米粒及制备方法和应用 |
CN106620680A (zh) * | 2017-01-03 | 2017-05-10 | 中国药科大学 | 一种树突细胞靶向的pH响应型DNA疫苗递送系统及制备方法 |
-
2019
- 2019-07-09 CN CN201910613285.9A patent/CN110404080B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1988000824A1 (en) * | 1986-07-28 | 1988-02-11 | Liposome Technology, Inc. | Liposomes with enhanced retention on mucosal tissue |
CN104193779A (zh) * | 2014-08-22 | 2014-12-10 | 四川大学 | 一种pH敏感磷脂药用材料及其制备方法与应用 |
CN105801668A (zh) * | 2016-04-01 | 2016-07-27 | 天津大学 | 一种寡聚精氨酸修饰的磷脂与其组装的纳米粒及制备方法和应用 |
CN106620680A (zh) * | 2017-01-03 | 2017-05-10 | 中国药科大学 | 一种树突细胞靶向的pH响应型DNA疫苗递送系统及制备方法 |
Non-Patent Citations (2)
Title |
---|
DIANA GUZMAN-VILLANUEVA ET AL.: "Influence of Triphenylphosphonium (TPP) Cation Hydrophobization with Phospholipids on Cellular Toxicity and Mitochondrial Selectivity", 《SOJ PHARM PHARM SCI》 * |
LEI JIANG ET AL.: "Dendritic phospholipid-based drug delivery systems", 《BIOMATER. SCI.》 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114042043A (zh) * | 2021-11-26 | 2022-02-15 | 河南中医药大学 | 一种雷公藤甲素线粒体靶向脂质体及其应用 |
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