CN108635593B - 一种e-选择素肽配体修饰的靶向热敏脂质体的制备和应用 - Google Patents
一种e-选择素肽配体修饰的靶向热敏脂质体的制备和应用 Download PDFInfo
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Abstract
本发明涉及一种E‑选择素多肽配体修饰的肿瘤靶向脂质体制备方法及应用,所述的靶向脂质体由磷脂酰乙醇胺‑聚乙二醇‑马来酰亚胺‑E选择素肽配体偶联物、二棕榈酰磷脂酰胆碱、培化磷脂酰乙醇胺、肉豆蔻酰溶血卵磷脂制备得到。其中涉及的偶联物DSPE‑PEG‑Mal‑8CR由E‑选择素肽配体8‑CR与磷脂酰乙醇胺‑聚乙二醇‑马来酰亚胺进行化学偶联合成,本发明制备的肿瘤靶向长循环热敏脂质体,包载不同的抗肿瘤药物后可以主动靶向于肿瘤新生血管并能够抑制肿瘤细胞的迁移,对应对肿瘤的转移和复发有着重要意义。
Description
技术领域
本发明属于生物医药制剂技术领域,涉及E选择素配体可与肿瘤细胞竞争结合E选择素从而阻断或抑制肿瘤细胞的迁移,特别涉及一种E-选择素肽类配体修饰的靶向热敏脂质体的制备和在抗肿瘤方面的应用。
技术背景
E-选择素可特异性识别肿瘤细胞表面的某些糖蛋白和糖脂分子的末端结构域,通过这种识别作用介导的E-选择素与肿瘤细胞间的作用,可使肿瘤细胞在内皮细胞黏附,继而随血流发生迁移,导致肿瘤细胞的扩散迁移。
E-选择素天然配体的结构特征仍未被完全阐明,有研究认为其特异性识别的天然配体可能包含糖类、肽类等多种结构类型,目前被广泛接受的E-选择素的主要天然配体结构为路易斯酸化的四聚糖结构sLex,理论上E-选择素的特异性配体可以作为靶向分子将其它抗肿瘤药物带到肿瘤部位,同时还可以发挥该配体对肿瘤细胞迁移的抑制作用,因此设计和合成与E-选择素结合的化合物可以起到肿瘤靶向和抑制肿瘤转移的双重作用。
纳米技术与生物技术的融合,使纳米生物技术在近几年获得了重要的进展。脂质体作为一种较成熟的纳米药物载体,具备一些独特的作用特点,脂质体是一种靶向药物载体,它可以通过纳米粒子的EPR效应被动靶向于肿瘤部位,减少药物在正常组织的分布,从而提高药物治疗效果,降低药物毒性作用。
目前各种不同方式修饰改良的脂质体不断被报道,例如制备热敏感的脂质体,聚乙二醇(PEG)修饰的脂质体等,这些脂质体通常具有较少的副作用,PEG化后还可显著延长血液循环时间和增加肿瘤组织的药物积聚。同时在脂质体表面还可以进行多种修饰,例如连接不同种类的靶向分子,进一步增加药物对肿瘤部位的靶向作用。但目前尚没有可以在上述基础上能同时解决肿瘤细胞转移问题的靶向脂质体,而肿瘤的转移又恰恰是目前肿瘤致死的最重要原因,本发明所设计制备的靶向脂质体不但可将药物靶向至肿瘤部位,还可以抑制由E-选择素所介导的肿瘤转移,因此具有明显的创新性和创造性。
发明内容
本发明的目的是提供用于治疗肿瘤及肿瘤转移的功能性脂质体的制备方法,本发明将E-选择素的肽配体修饰至DSPE-PEG-Mal表面,然后用于制备一种靶向脂质体,可以用作药物载体包载各种不同的药物,如以紫杉醇为药物模型,将热敏脂质体与主动靶向技术相结合,提高热敏脂质体的靶向性,降低药物的毒副作用,从而达到增强热敏脂质体的抗肿瘤效果的目的,同时抑制肿瘤的转移。
本发明是通过以下技术方案实现的:
一种E-选择素的肽配体修饰的靶向脂质体载药系统,含有磷脂酰乙醇胺-聚乙二醇-马来酰亚胺-E-选择素肽配体偶联物、二棕榈酰磷脂酰胆碱、培化磷脂酰乙醇胺、肉豆蔻酰溶血卵磷脂。
而且,所述E-选择素肽配体在肽配体基础上在其氨基端或羧基端分别添加半胱氨酸作为连接桥。
而且,所述E-选择素肽配体包括额瓶体的各种反序形式和D型氨基酸序列形式,CIELLQAR,IELLQARC,CRAQLLEI,RAQLLEIC,(D)-CIELLQAR,(D)-IELLQARC,(D)-CRAQLLEI,(D)-RAQLLEIC。
而且,其中所述抗肿瘤药物为紫杉醇、阿霉素、多西紫杉醇、多柔比星或奥沙利铂。
而且,所述二棕榈酰磷脂酰胆碱、培化磷脂酰乙醇胺、肉豆蔻酰溶血卵磷脂、磷脂酰乙醇胺-聚乙二醇-马来酰亚胺-E-选择素肽配体偶联物质量比为80~85:5~10:1~5:5~10。
而且,药物与磷脂的质量比为1:25~30。
而且,所述磷脂酰乙醇胺-聚乙二醇-马来酰亚胺-E-选择素肽配体偶联物的合成方法如下:
①按照传统固相合成技术合成E-选择素肽配体;
②多肽与DSPE-PEG-Mal偶联,方法如下:
而且,步骤如下:
①将二棕榈酰磷脂酰胆碱、培化磷脂酰乙醇胺、肉豆蔻酰溶血卵磷脂、紫杉醇、DSPE-PEG-Mal-8CR按处方量溶解于氯仿中形成溶液,充分溶解后于旋转蒸发仪上50~55℃条件下去除有机溶剂;
②将上述样品放于真空干燥箱真空干燥2~4小时;
③加入pH=7.2的柠檬酸/柠檬酸钠缓冲溶液,于55~60℃条件下水化溶解30min;
④用细胞破碎仪超声破碎6~10min,滤膜过滤,得到E-选择素配体肽修饰的靶向脂质体。
E-选择素的肽配体修饰的靶向脂质体载药系统在制备抗肿瘤及抗肿瘤转移药物中的应用。
与现有技术相比,本发明的有益效果:
1、本发明所涉及的脂质体载药系统制备简单、方便,适用于多种抗肿瘤药物。
2、热敏脂质体在局部加热条件下可定点大量释放所载药物,同时E-选择素配体CIELLQAR又可主动靶向肿瘤血管,起到协同肿瘤靶向治疗,最重要的是E-选择素肽配体CIELLQAR还可以抑制肿瘤细胞与血管内皮细胞之间的黏附作用,进而能够抑制肿瘤的转移。
3、本发明通过处方工艺条件的优化,制备得到E-选择素肽配体修饰的靶向脂质体具有包封率高、粒径分布均匀,制剂可稳定贮存等优点。
附图说明
图1为本发明实施例制备的包载紫杉醇的E-选择素肽配体修饰的靶向脂质体制剂(脂质体F)的成品图。
图2为原料DSPE-PEG-Mal(左侧峰)和产物DSPE-PEG-Mal-8CR(右侧峰)的MALDI-TOF对比图谱。
图3为本发明实施例包载紫杉醇的E-选择素肽配体修饰的靶向脂质体(脂质体F)用动态光散射(DLS)测得的粒径分布图。
图4为本发明实施例包载紫杉醇的E-选择素肽配体修饰的靶向脂质体(脂质体F)用动态光散射(DLS)测得的电势图。
图5为各种脂质体的体外抗粘附实验结果。
具体的实施方式
为了理解本发明,下面结合实施例对本发明作进一步说明:下述实施例是说明性的,不是限定性的,不能以下述实施例来限定本发明的保护范围。
本发明提供的E-选择素肽配体修饰的靶向脂质体载药系统,其组成包含:磷脂酰乙醇胺-聚乙二醇-马来酰亚胺-E-选择素肽配体偶联物、二棕榈酰磷脂酰胆碱、培化磷脂酰乙醇胺、肉豆蔻酰溶血卵磷脂、有机溶剂和蒸馏水。
所述的E-选择素肽配体,在IELLQAR这一文献已报道的肽配体基础上在其氨基端或羧基端分别添加半胱氨酸作为连接桥,以及该肽的各种反序形式和D型氨基酸序列形式。如:CIELLQAR,IELLQARC,CRAQLLEI,RAQLLEIC,(D)-CIELLQAR,(D)-IELLQARC,(D)-CRAQLLEI,(D)-RAQLLEIC等。该类肽配体能够特异性地与血管内皮细胞上的E-选择素受体结合,并阻断E-选择素介导的肿瘤细胞的转移。(参考文件APeptideMimicofE-SelectinLigandInhibitsSialyl LewisX-dependentLungColonizationofTumorCells,CancerResearch,2000,60(2),450-456.第二篇:SialylLewisX-dependentlungcolonizationofB16melanomacellsthroughaselectin-likeendothelialreceptordistinctfromE-orP-selectin,CancerResearch,2002,62(62),4194-4198.)
其中,磷脂酰乙醇胺-聚乙二醇-马来酰亚胺-E-选择素肽配体偶联物的合成方法如下:
①按照传统固相合成技术合成E-选择素肽配体,例如CIELLQAR。
②多肽与DSPE-PEG-Mal偶联,方法如下:
其中所述抗肿瘤药物可以为脂溶或水溶等不同性质的各种抗肿瘤药物,例如紫杉醇、阿霉素、多西紫杉醇、多柔比星、奥沙利铂等。
所述二棕榈酰磷脂酰胆碱、培化磷脂酰乙醇胺、肉豆蔻酰溶血卵磷脂、DSPE-PEG-Mal-8CR质量比为80~85:5~10:1~5:5~10。药物与磷脂的质量比为1:25~30。磷脂与水的质量比为100:3。
其中,E-选择素肽配体修饰的靶向脂质体的制备方法,包括以下步骤:
①将二棕榈酰磷脂酰胆碱、培化磷脂酰乙醇胺、肉豆蔻酰溶血卵磷脂、紫杉醇、DSPE-PEG-Mal-8CR按处方量溶解于氯仿中形成溶液,充分溶解后于旋转蒸发仪上50~55℃条件下去除有机溶剂。
②将上述样品放于真空干燥箱真空干燥2~4小时。
③加入pH=7.2的柠檬酸/柠檬酸钠缓冲溶液,于55~60℃条件下水化溶解30min。
④用细胞破碎仪超声破碎6~10min,0.22μm滤膜过滤,得到E-选择素配体肽修饰的靶向脂质体,所制备的靶向药物制剂的粒径为85~120nm,Zeta电位为-28~-10mV。
针对上述合成方法,本申请提供具体的操作方式,如以下实施例,首先采用经典固相Fmoc策略,使用2-氯三苯甲基氯树脂(2-ChlorotritylChlorideResin)合成获得所需的E选择素配体CIELLQAR(8-CR)。
实施例1
E-选择素肽配体8-CR与DSPE-PEG2000-Mal的连接。
称取18.9mg的8-CR溶解于4mlHEPES缓冲溶液(20mMHEPES,10mM EDTA,pH=6.5)中,称取31.3mgDSPE-PEG2000-Mal溶解于4ml氯仿中,充分溶解后于旋转蒸发仪上50℃条件下去除有机溶剂。然后将其用HEPES缓冲溶液于55℃条件下水化溶解15min,将其加入到多肽溶液中,即8-CR与DSPE-PEG2000-Mal的摩尔比为2∶1,在氩气保护下,室温搅拌48小时。反应结束后将上述体系置于截留分子量为3000的透析袋中透析48小时,经冷冻干燥得到DSPE-PEG2000-Mal-8CR。
实施例2空白脂质体的制备。
普通空白脂质体的制备。
分别称取二棕榈酰磷脂酰胆碱、培化磷脂酰乙醇胺、肉豆蔻酰溶血卵磷脂、DSPE-PEG2000-Mal各166mg、18mg、6mg、10mg溶解于氯仿中形成溶液,充分溶解后于旋转蒸发仪上(55℃,60r/min)条件下去除有机溶剂,然后将上述脂质体放于真空干燥箱真空干燥3小时。在将干燥后的脂质体加6mlPH=7.2的柠檬酸/柠檬酸钠缓冲溶液于旋转蒸发仪(58℃,100r/min)水化溶解25min。将上述脂质体溶液用细胞破碎仪超声破碎5min,过0.22μm膜,得到普通空白脂质体G。
靶向空白脂质体的制备。
分别称取二棕榈酰磷脂酰胆碱、培化磷脂酰乙醇胺、肉豆蔻酰溶血卵磷脂、DSPE-PEG2000-Mal-8CR各166mg、18mg、6mg、10mg溶解于氯仿中形成溶液,充分溶解后于旋转蒸发仪上(55℃,60r/min)条件下去除有机溶剂,然后将上述脂质体放于真空干燥箱真空干燥3小时。在将干燥后的脂质体加6ml pH=7.2的柠檬酸/柠檬酸钠缓冲溶液于旋转蒸发仪(58℃,100r/min)水化溶解25min。将上述脂质体溶液用细胞破碎仪超声破碎5min,过0.22μm膜,得到靶向空白脂质体D。
实施例3载药脂质体的制备。
普通载药脂质体的制备。
分别称取二棕榈酰磷脂酰胆碱、培化磷脂酰乙醇胺、肉豆蔻酰溶血卵磷脂、DSPE-PEG2000-Mal、紫杉醇各166mg、18mg、6mg、10mg、8mg溶解于氯仿中形成溶液,充分溶解后于旋转蒸发仪上(55℃,60r/min)条件下去除有机溶剂,然后将上述脂质体放于真空干燥箱真空干燥3小时。在将干燥后的脂质体加6mlpH=7.2的柠檬酸/柠檬酸钠缓冲溶液于旋转蒸发仪(58℃,100r/min)水化溶解25min。将上述脂质体溶液用细胞破碎仪超声破碎5min,过0.22μm膜,得到普通载药脂质体E。
靶向载药脂质体的制备。
分别称取二棕榈酰磷脂酰胆碱、培化磷脂酰乙醇胺、肉豆蔻酰溶血卵磷脂、DSPE-PEG2000-Mal-8CR、紫杉醇各166mg、18mg、6mg、10mg、8mg溶解于氯仿中形成溶液,充分溶解后于旋转蒸发仪上(55℃,60r/min)条件下去除有机溶剂,然后将上述脂质体放于真空干燥箱真空干燥3小时。在将干燥后的脂质体加6mlpH=7.2的柠檬酸/柠檬酸钠缓冲溶液于旋转蒸发仪(58℃,100r/min)水化溶解25min。将上述脂质体溶液用细胞破碎仪超声破碎5min,过0.22μm膜,得到靶向载药脂质体F。
实施例4
脂质体包封率、粒径和电位的测定。
包封率测定:利用紫杉醇难溶于水的特性,采用高速离心法去除未载入脂质体的游离紫杉醇。用移液器枪移取靶向载药脂质体溶液200微升,蒸馏水稀释后加入5%SDS破乳剂破乳,涡旋混匀后用高效液相分析总的紫杉醇浓度(w总)。另移取200微升靶向载药脂质体溶液,蒸馏水稀释后离心,沉淀用乙腈溶解,高效液相分析游离的紫杉醇浓度(w游)。药物包封率计算公式如下:
EE=1-(W游/W总)×100%
粒径和电位的测定:取20微升脂质体溶液,加蒸馏水稀释到1.6ml,利用MalvernZetasizerNanoZS90纳米粒径电位分析仪测定脂质体的粒径、多分散系数和Zeta电位。脂质体粒径分布均匀,粒径大小在90nm~100nm之间。各脂质体都带负电,说明脂质体纳米粒子之间具有适量的静电排斥力,贮存稳定性更好,不易发生聚沉。不同脂质体的粒径、多分散系数(PDI)和Zeta电位以及包封率的测定结果见下表。
实施例5
脂质体体外抗粘附实验测定方法
内皮细胞HUVEC细胞株和人急性白血病细胞系HL-60均购买于ATCC,于37℃,5%CO2饱和湿度的培养中培养,培养基分别为10%胎牛血清,100μg/mL青霉素和100μg/mL链霉素的RPMI1640和F-12,实验所用为对数生长期细胞。多聚赖氨酸(20μg/ml)(上海源叶生物科技有限公司)包被黑色96孔细胞培养板(CorningIncorporatedcostar3603),37℃孵育12h。内皮细胞HUVEC以1×105cell/well的浓度接种于黑色96孔细胞培养板,37℃孵育36h后,移去细胞培养板中的培养基,实验组加入含有20ng/mlTNF-α的F-12培养基,对照组加入未添加TNF-α的F-12培养基,37℃孵育6h使其表达E-selectin。同时,收集对数生长期的人急性白血病细胞HL-60(1×105cell/well),1000rpm/min,离心5min。加入5mlPBS清洗,1000rpm/min,离心5min,弃上清,加入100μl1640培养基吹悬,加入20μM钙黄绿素CalceinAM(Lifetechnologies),避光,37℃孵育45min后,1000rpm/min,离心5min,弃上清,加入1mlPBS清洗,1000rpm/min,离心5min,弃上清,加入1640培养基将沉淀吹悬到所需体积,以1×107cell/well的密度接种于加有HUVEC细胞的96孔细胞培养板中后立即加入不同浓度的候选脂质体,每个浓度设置三个重复孔,对照组只加入缓冲液,37℃避光孵育30min后加入PBS清洗细胞培养板三次以去除未与HUVEC细胞结合的HL-60细胞。每孔加入100μl透膜液(BIOSHARP),用Synergy4多功能微孔板检测仪(Biotek)测荧光(Ex,485nm;Em,528nm)。脂质体的黏附抑制作用与荧光强度成反比例关系,荧光值越低,说明化合物的黏附抑制效果越强。
Claims (4)
1.一种E-选择素的肽配体修饰的靶向脂质体载药系统,其特征在于:含有磷脂酰乙醇胺-聚乙二醇-马来酰亚胺-E-选择素肽配体偶联物、二棕榈酰磷脂酰胆碱、培化磷脂酰乙醇胺、肉豆蔻酰溶血卵磷脂和抗肿瘤药物;所述二棕榈酰磷脂酰胆碱、培化磷脂酰乙醇胺、肉豆蔻酰溶血卵磷脂、磷脂酰乙醇胺-聚乙二醇-马来酰亚胺-E-选择素肽配体偶联物质量比为80~85:5~10:1~5:5~10;所述抗肿瘤药物为紫杉醇;
所述E-选择素肽配体在肽配体基础上在其氨基端或羧基端分别添加半胱氨酸作为连接桥;所述E-选择素肽配体为CIELLQAR;
载药系统的制备方法如下:
①将二棕榈酰磷脂酰胆碱、培化磷脂酰乙醇胺、肉豆蔻酰溶血卵磷脂、紫杉醇、DSPE-PEG-Mal-CIELLQAR按处方量溶解于氯仿中形成溶液,充分溶解后于旋转蒸发仪上50~55℃条件下去除有机溶剂;
②将上述样品放于真空干燥箱真空干燥2~4小时;
③加入pH=7.2的柠檬酸/柠檬酸钠缓冲溶液,于55~60℃条件下水化溶解30min;
④用细胞破碎仪超声破碎6~10min,滤膜过滤,得到E-选择素肽配体修饰的靶向脂质体。
2.根据权利要求1所述的E-选择素的肽配体修饰的靶向脂质体载药系统,其特征在于:药物与磷脂的质量比为1:25~30。
4.权利要求1所述的E-选择素的肽配体修饰的靶向脂质体载药系统在制备抗肿瘤药物中的应用。
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CN102552144A (zh) * | 2012-01-13 | 2012-07-11 | 北京大学 | 一种靶向热敏脂质体 |
CN107468651A (zh) * | 2017-08-02 | 2017-12-15 | 中国药科大学 | 一种包载基质金属蛋白酶抑制剂的温度响应型纳米脂质体的制备方法 |
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101855237A (zh) * | 2007-11-05 | 2010-10-06 | 上海交通大学 | 肽配体定向药物递送 |
CN102552144A (zh) * | 2012-01-13 | 2012-07-11 | 北京大学 | 一种靶向热敏脂质体 |
CN107468651A (zh) * | 2017-08-02 | 2017-12-15 | 中国药科大学 | 一种包载基质金属蛋白酶抑制剂的温度响应型纳米脂质体的制备方法 |
Non-Patent Citations (2)
Title |
---|
A Peptide Mimic of E-Selectin Ligand Inhibits Sialyl Lewis X-dependent Lung Colonization of Tumor Cells;Michiko N. Fukuda, et al.;《CANCER RESEARCH》;20000115;第60卷;450-456 * |
Effect of sialyl Lewis X-glycoliposomes on the inhibition of E-selectin-mediated tumour cell adhesion in vitro;Reinhard Zeisig,et al.;《Biochimica et Biophysica Acta》;20041231;第1660卷;31-40 * |
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