脑靶向脂质体构建及其靶分子Ac_4MAN体外结构稳定性研究
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摘要
目的构建修饰的Ac_4MAN(p-羧基苯-α-D-乙酰甘露糖)脑靶向脂质体,并探讨Ac_4MAN结构稳定性。方法将Ac_4MAN与猪肝酯酶(PLE)于37℃、pH 7.4条件下孵育,于不同时间点取样;采用高效液相色谱(HPLC)和超高效液相色谱-质谱(UPLC-MS)联用技术对其进行分析。制备Ac_4MAN脂质体并对其进行表征,用流式细胞仪检测脑胶质瘤细胞U87摄取Ac_4MAN脂质体和Ac_4MAN脂质体加PLE的情况。结果 HPLC结果显示,孵育0～3 h,Ac_4MAN峰面积从578.4减少到37.9;孵育4～7 h,Ac_4MAN峰面积接近于0;孵育0～7 h,生成的MAN(p-氨基苯-α-D-吡喃甘露糖苷)的峰面积从0增长到363.0,表明随孵育时间延长,Ac_4MAN逐渐减少,MAN逐渐增多。结论本研究制备的Ac_4MAN脂质体粒径约为(120.3±2.0)nm,电位约为(-15.76±1.23)mV。Ac_4MAN脂质体中加入酶后,U87细胞对其摄取能力有一定的增强。
Objective To construct a modified Ac_4 MAN(p-carboxybenzene-α-D-acetylmannose) brain-targeted liposome and investigate the structural stability of Ac_4 MAN. Methods Ac_4 MAN was incubated with porcine liver esterase(PLE) at 37°C, pH 7.4 and sampled at different time points. High performance liquid chromatography(HPLC) and ultra performance liquid chromatography-mass spectrometry(UPLC-MS) were used to analyze it. Ac_4 MAN liposomes were prepared and characterized. Flow cytometry was used to detect the uptake of Ac_4 MAN liposomes and Ac_4 MAN liposomes plus PLE by glioma cells U87. Results The results of HPLC showed that the area of Ac_4 MAN decreased from578.4 to 37.9 after incubation for 0-3 h. The area of Ac_4MAN peak was close to 0 after incubation for 4-7 h. The peak area of the formed MAN(p-aminobenzene-α-D-pyran mannoside) increased from 0 to 363.0, indicating that with the prolongation of incubation time, Ac_4 MAN gradually decreased and MAN gradually increased. Conclusion The Ac_4 MAN liposome prepared in this study has a particle size of( 120. 3 ± 2. 0) nm and a potential of(-15.76 ±1.23) m V. The uptake of AC4 MAN liposomes by U87 cells is enhanced after adding enzymes.
Objective To construct a modified Ac_4 MAN(p-carboxybenzene-α-D-acetylmannose) brain-targeted liposome and investigate the structural stability of Ac_4 MAN. Methods Ac_4 MAN was incubated with porcine liver esterase(PLE) at 37°C, pH 7.4 and sampled at different time points. High performance liquid chromatography(HPLC) and ultra performance liquid chromatography-mass spectrometry(UPLC-MS) were used to analyze it. Ac_4 MAN liposomes were prepared and characterized. Flow cytometry was used to detect the uptake of Ac_4 MAN liposomes and Ac_4 MAN liposomes plus PLE by glioma cells U87. Results The results of HPLC showed that the area of Ac_4 MAN decreased from578.4 to 37.9 after incubation for 0-3 h. The area of Ac_4MAN peak was close to 0 after incubation for 4-7 h. The peak area of the formed MAN(p-aminobenzene-α-D-pyran mannoside) increased from 0 to 363.0, indicating that with the prolongation of incubation time, Ac_4 MAN gradually decreased and MAN gradually increased. Conclusion The Ac_4 MAN liposome prepared in this study has a particle size of( 120. 3 ± 2. 0) nm and a potential of(-15.76 ±1.23) m V. The uptake of AC4 MAN liposomes by U87 cells is enhanced after adding enzymes.
引文
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[3]Shukla GH,Kour Khera A,Kumar S,et al.Therapeutic potential,challenges and future perspective of cancer stem cells in translational oncology:a critical review[J].Curr Stem Cell Rest,2017,12(3):207-224.
[4]Bar-Zeev M,Livney YD,Assaraf YG.Targeted nanomedicine for cancer therapeutics:Towards precision medicine overcoming drug resistance[J].Drug Resist Update,2017,31:15-30.
[5]Mujokoro B,Adabi M,Sadroddiny E,et al.Nano-structures mediated co-delivery of therapeutic agents for glioblastoma treatment:A review[J].Mater Sci Eng C Mater Biol Appl,2016,69:1092-1102.
[6]Riaz MK,Riaz MA,Zhang X,et al.Surface functionalization and targeting strategies of liposomes in solid tumor therapy:A revie[J].Int J Mol Sci,2018,19(1):195-212.
[7]Pinto MP,Arce M,Yameen B,et al.Targeted brain delivery nanoparticles for malignant gliomas[J].Nanomedicine(Lond),2017,12(1):59-72.
[8]Maherani BE,Arab-Tehrany M,Mozafari R,et al.Liposomes:a review of manufacturing techniques and targeting strategies[J].Curr Nano Sci,2011,7(3):436-452.
[9]Siafaka PI,Ustundag Okur N,Karavas E,et al.Surface modified multifunctional and stimuli responsive nanoparticles for drug targeting:current status and uses[J].Int JMol Sci,2016,17(9):1440.
[10]Szablewski L.Glucose Transporters in Brain:In Health and in Alzheimer′s Disease[J].J Alzheimers Dis,2017,55(4):1307-1320.
[11]Hao ZF,Cui YX,Li MH,et al.Liposomes modified with P-aminophenyl-alpha-D-manno-pyranoside:a carrier for targeting cerebral functional regions in mice[J].Eur J Pharm Biopharm,2013,84(3):505-516.
[12]Nishioka T,Oda Y,Seino Y,et al.Distribution of the glucose transporters in human brain tumors[J].Cancer Res,1992,52(14):3972-3979.
[13]Du D,Chang N,Sun S,et al.The role of glucose transporters in the distribution of p-aminophenyl-alpha-D-mannopyranoside modified liposomes within mice brain[J].J Control Release,2014,182:99-110.
[14]Miranda A,Blanco-Prieto MJ,Sousa J,et al.Breaching barriers in glioblastoma.PartⅡ:Targeted drug delivery and lipid nanoparticles[J].Int J Pharm,2017,531(1):389-410.
[15]Woodle MC,Newman MS,Working PK.Biological properties of sterically stabilized liposomes,in Stealth liposomes[J].CRC Press,2018:123-138.
[16]Pradhan P,Banerjee R,Bahadur D,et al.Targeted Magnetic Liposomes Loaded with Doxorubicin,in Liposomes[J].Springer,2017:257-272.
[17]Weissig V.Liposomes came first:The early history of liposomology,in Liposomes[J].Springer,2017:1-15.
[18]Bnyan R,Khan I,Ehtezazi T,et al.Surfactant Effects on Lipid-Based Vesicles Properties[J].J Pharm Sci,2018,107(5):1237-1246.
[19]Mu LM,Ju RJ,Liu R,et al.Dual-functional drug liposomes in treatment of resistant cancers[J].Adv Drug Deliver Rev,2017,115:46-56.
[20]Jurcovicova J.Glucose transport in brain-effect of inflammation[J].Endocr Regul,2014,48(1):35-48.