AEYLR小肽修饰的紫杉醇纳米结构脂质载体的制备及抗肿瘤效果评价
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摘要
目的:制备序列为丙氨酸-谷氨酸-酪氨酸-亮氨酸-精氨酸(简称为"AEYLR")的小肽修饰的紫杉醇(PTX)纳米结构脂质载体(A-P-NLC),并对其体内外抗肿瘤效果进行评价。方法:采用熔融乳化-低温固化法制备纳米结构脂质载体(NLC)、PTX纳米结构脂质载体(P-NLC)和A-P-NLC,表征其外观形态、粒径、多分散指数(PDI)、Zeta电位,并检测其包封率、载药量及体外释放度;以NCI-H1299细胞和S180细胞为对象,采用CCK-8法对游离PTX、P-NLC、A-P-NLC(0.44～44.00μg/mL,以PTX计)的细胞抑制作用进行考察,并计算其半数抑制浓度(IC50);以S180荷瘤小鼠为模型动物,对游离PTX、P-NLC、A-P-NLC(5 mg/kg,以PTX计)的抑瘤效果进行评价。结果:P-NLC和A-P-NLC外观均呈类圆形、分布均匀;A-P-NLC的粒径、PDI、Zeta电位分别为(43.92±0.76)nm、0.203±0.034、(-19.77±1.16)m V,较P-NLC有所增加;A-P-NLC的包封率、载药量分别为(95.71±0.68)%、(1.97±0.25)%,较P-NLC有所降低;A-P-NLC在48 h内累积释放百分率达(35.17±2.08)%,较游离PTX表现出明显的缓释作用,且比P-NLC的释放更缓慢。与游离PTX和P-NLC比较,相同质量浓度的A-P-NLC对NCI-H1299细胞和S180细胞的抑制率大部分均显著升高,IC50值均显著降低;A-P-NLC给药处理的S180荷瘤小鼠无死亡现象,一般状态良好,且瘤体积显著缩小、瘤质量显著降低、瘤质量抑制率显著升高(P<0.05或P<0.01)。结论:A-P-NLC具有明显的缓释作用,其对NCI-H1299细胞和S180细胞的体外抑制作用以及对小鼠S180实体瘤的抑制作用均优于游离PTX和P-NLC,且毒性有所降低。
OBJECTIVE: To prepare Paclitaxel(PTX)nanostructured lipid carriers(NLC) modified by small peptide alanine-glutamic acid-tyrosine-leucine-arginine(AEYLR),and to evaluate its anti-tumor effect in vitro and in vivo. METHODS:NLC,PTX-NLC(P-NLC) and AEYLR modified P-NLC(A-P-NLC) were prepared by emulsion evaporation-low temperature solidification curing method. Its appearance,particle size,multi-dispersion index(PDI) and Zeta potential were characterized,encapsulation rate,drug loading and in vitro drug release were detected respectively. Using NCI-H1299 and S180 cells as objects,CCK-8 method was adopted to investigate inhibitory effects of free PTX,P-NLC and A-P-NLC(0.44-44.00 μg/mL,by PTX)to those cells. The half inhibition concentration(IC50)was calculated. Using S180 tumor-bearing mice as model animal,anti-tumor effects of free PTX,P-NLC and A-P-NLC(5 mg/kg,by PTX)were evaluated. RESULTS:P-NLC and A-P-NLC were round-like and dispersed evenly. The particle size,PDI and Zeta potential of A-P-NLC were(43.92±0.76)nm,0.203±0.034 and(-19.77±1.16)mV,which were all increased to certain extent,compared with P-NLC. The encapsulation efficiency and drug loading of A-P-NLC were(95.71 ± 0.68)% and(1.97 ± 0.25)%,which were both decreased to certain extent,compared with P-NLC. The cumulative release rate of A-P-NLC was(35.17±2.08)% within 48 h,showing significant sustained-release effect compared with free PTX;the release of A-P-NLC was slower than P-NLC. Compared with free PTX and P-NLC,inhibitory rates of same concentration of A-P-NLC to NCI-H1299 cells and S180 cells were almost increased significantly,while IC50 values were all decreased significantly. There was no death in S180 tumor-bearing mice treated with A-P-NLC and the general condition was good;the volume of tumors was significantly reduced,the mass of tumors was significantly reduced,and the inhibition rate of tumors was significantly increased(P<0.05 or P<0.01). CONCLUSIONS:A-P-NLC has significantly sustained-release effects;its inhibitory rate to NCI-H1299 cells and S180 cells in vitro,and its inhibitory effects on S180 solid tumor in mice are all better than free PTX and P-NLC,while the toxicity is decreased to certain extent.
OBJECTIVE: To prepare Paclitaxel(PTX)nanostructured lipid carriers(NLC) modified by small peptide alanine-glutamic acid-tyrosine-leucine-arginine(AEYLR),and to evaluate its anti-tumor effect in vitro and in vivo. METHODS:NLC,PTX-NLC(P-NLC) and AEYLR modified P-NLC(A-P-NLC) were prepared by emulsion evaporation-low temperature solidification curing method. Its appearance,particle size,multi-dispersion index(PDI) and Zeta potential were characterized,encapsulation rate,drug loading and in vitro drug release were detected respectively. Using NCI-H1299 and S180 cells as objects,CCK-8 method was adopted to investigate inhibitory effects of free PTX,P-NLC and A-P-NLC(0.44-44.00 μg/mL,by PTX)to those cells. The half inhibition concentration(IC50)was calculated. Using S180 tumor-bearing mice as model animal,anti-tumor effects of free PTX,P-NLC and A-P-NLC(5 mg/kg,by PTX)were evaluated. RESULTS:P-NLC and A-P-NLC were round-like and dispersed evenly. The particle size,PDI and Zeta potential of A-P-NLC were(43.92±0.76)nm,0.203±0.034 and(-19.77±1.16)mV,which were all increased to certain extent,compared with P-NLC. The encapsulation efficiency and drug loading of A-P-NLC were(95.71 ± 0.68)% and(1.97 ± 0.25)%,which were both decreased to certain extent,compared with P-NLC. The cumulative release rate of A-P-NLC was(35.17±2.08)% within 48 h,showing significant sustained-release effect compared with free PTX;the release of A-P-NLC was slower than P-NLC. Compared with free PTX and P-NLC,inhibitory rates of same concentration of A-P-NLC to NCI-H1299 cells and S180 cells were almost increased significantly,while IC50 values were all decreased significantly. There was no death in S180 tumor-bearing mice treated with A-P-NLC and the general condition was good;the volume of tumors was significantly reduced,the mass of tumors was significantly reduced,and the inhibition rate of tumors was significantly increased(P<0.05 or P<0.01). CONCLUSIONS:A-P-NLC has significantly sustained-release effects;its inhibitory rate to NCI-H1299 cells and S180 cells in vitro,and its inhibitory effects on S180 solid tumor in mice are all better than free PTX and P-NLC,while the toxicity is decreased to certain extent.
引文
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[11]靳彩玲,赵树鹏,张敏,等.转铁蛋白修饰紫杉醇脂质体的制备及其抑瘤作用[J].中国药房,2016,27(1):44-47.
[12]WANG W,CHEN L,HUANG X,et al.Preparation and characterization of minoxidil loaded nanostructured lipid carrier[J].AAPS PharmSciTech,2016,18(2):1-8.
[13]CAO J,HOU D,LU J,et al.Anti-tumor activity of exopolysaccharide from Rhizopus nigricans Ehrenb on S180tumor-bearing mice[J].Bioorg Med Chem Lett,2016,26(8):2098-2104.
[14]姜一朴,滕晋莹,胡金芳,等.小鼠H22、S180腹水模型临床检查特征研究[J].药物评价研究,2018,41(8):1386-1390.
[15]李学涛,程岚,姜英,等.长春碱亲水基修饰阳离子脂质体对荷瘤小鼠的抗肿瘤作用研究[J].中国药房,2015,26(31):4339-4341.
[16]OLERILE LD,LIU Y,BO Z,et al.Near-infrared mediated quantum dots and paclitaxel co-loaded nanostructured lipid carriers for cancer theragnostic[J].Colloids Surf BBiointerfaces,2017.DOI:10.1016/j.colsurfb.2016.11.032.
[17]LIAO H,ZHAO X,QU J,et al.Matrine suppresses invasion and metastasis of NCI-H1299 cells by enhancing microRNA-133a expression[J].Int J Clin Exp Med,2015,8(7):10714-10722.
[18]WANG J,CHEN S,XU S,et al.In vivo induction of apoptosis by fucoxanthin,a marine carotenoid,associated with down-regulating STAT3/EGFR signaling in sarcoma 180(S180)xenografts-bearing mice[J].Mar Drugs,2012,10(9):2055-2068.
[2]TUTEJA M,KANG M,LEAL C,et al.Nanoscale partitioning of paclitaxel in hybrid lipid-polymer membranes[J].Analyst,2018,143(16):3808-3813.
[3]ZHAI J,LUWOR RB,AHMED N,et al.Paclitaxel-loaded self-assembled lipid nanoparticles as targeted drug delivery systems for the treatment of aggressive ovarian cancer[J].ACS Appl Mater Interfaces,2018,10(30):25174-25185.
[4]王子琪,王狄狮,李馨儒,等.紫杉醇的pH敏感聚合物胶束的制备及其体外释放研究[J].中国新药杂志,2018,27(4):459-464.
[5]PARDEIKE J,HOMMOSS A,MüLLER RH.Lipid nanoparticles(SLN,NLC)in cosmetic and pharmaceutical dermal products[J].Int J Pharm,2009,366(1/2):170-184.
[6]CHAKRABORTY S,SHUKLA D,MISHRA B,et al.Lipid:an emerging platform for oral delivery of drugs with poor bioavailability[J].Eur J Pharm Biopharm,2009,73(1):1-15.
[7]FENG L,MUMPER RJ.A critical review of lipid-based nanoparticles for taxane delivery[J].Cancer Lett,2013,334(2):157-175.
[8]LIN X,GAO R,ZHANG Y,et al.Lipid nanoparticles for chemotherapeutic applications:strategies to improve anticancer efficacy[J].Expert Opin Drug Deliv,2012,9(7):767-781.
[9]HAN C,YUE L,TAI L,et al.A novel small peptide as an epidermal growth factor receptor targeting ligand for nanodelivery in vitro[J].Int J Nanomedicine,2013.DOI:10.2147/IJN.S43627.
[10]HAN C,LI Y,SUN M,et al.Small peptide-modified nanostructured lipid carrier distribution and targeting to EGFR overexpressing tumor in vivo[J].Artif Cells Nanomed Biotechnol,2014,42(3):161-166.
[11]靳彩玲,赵树鹏,张敏,等.转铁蛋白修饰紫杉醇脂质体的制备及其抑瘤作用[J].中国药房,2016,27(1):44-47.
[12]WANG W,CHEN L,HUANG X,et al.Preparation and characterization of minoxidil loaded nanostructured lipid carrier[J].AAPS PharmSciTech,2016,18(2):1-8.
[13]CAO J,HOU D,LU J,et al.Anti-tumor activity of exopolysaccharide from Rhizopus nigricans Ehrenb on S180tumor-bearing mice[J].Bioorg Med Chem Lett,2016,26(8):2098-2104.
[14]姜一朴,滕晋莹,胡金芳,等.小鼠H22、S180腹水模型临床检查特征研究[J].药物评价研究,2018,41(8):1386-1390.
[15]李学涛,程岚,姜英,等.长春碱亲水基修饰阳离子脂质体对荷瘤小鼠的抗肿瘤作用研究[J].中国药房,2015,26(31):4339-4341.
[16]OLERILE LD,LIU Y,BO Z,et al.Near-infrared mediated quantum dots and paclitaxel co-loaded nanostructured lipid carriers for cancer theragnostic[J].Colloids Surf BBiointerfaces,2017.DOI:10.1016/j.colsurfb.2016.11.032.
[17]LIAO H,ZHAO X,QU J,et al.Matrine suppresses invasion and metastasis of NCI-H1299 cells by enhancing microRNA-133a expression[J].Int J Clin Exp Med,2015,8(7):10714-10722.
[18]WANG J,CHEN S,XU S,et al.In vivo induction of apoptosis by fucoxanthin,a marine carotenoid,associated with down-regulating STAT3/EGFR signaling in sarcoma 180(S180)xenografts-bearing mice[J].Mar Drugs,2012,10(9):2055-2068.