艾塞那肽聚乳酸-羟基乙酸共聚物纳米粒制备及体外评价
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摘要
目的:制备艾塞那肽聚乳酸-羟基乙酸共聚物[poly(lactic-co-glycolic acid),PLGA]纳米粒并对其进行体外评价及表征。方法:通过正交实验设计优化法筛选艾塞那肽PLGA纳米粒的处方及工艺,并通过动态光散射和透射电镜对其进行了表征,通过体外模拟胃肠液和Caco-2细胞模型对其稳定性和细胞摄取及转运进行评价。结果:通过正交设计最终确定了最佳处方工艺:内水相和油相的比例为1∶5,超声功率200 W,超声时间4 min,聚乙烯醇(polyvinyl alcohol,PVA)浓度为2%,初乳与外水相体积比为1∶20。所制得纳米粒粒径为116.2 nm,透射电镜显示PLGA纳米粒为类圆形球状纳米粒,表面规则。体外研究结果表明,PLGA纳米粒可延缓艾塞那肽在胃肠液中的释放,同时也明显增加了其稳定性(P<0.05),在人工胃液和人工肠液中孵育4 h后,仍分别有约50%和80%的保留。细胞实验表明,PLGA纳米粒的摄取和转运分别是对照组的7倍和5倍之多(P<0.05)。结论:PLGA纳米粒可作为艾塞那肽潜在的高效口服递药系统。
Objective:To prepare exenatide-loaded poly(lactic-co-glycolic acid)(PLGA)nanoparticles,and to evaluate its in vitro effect and characteristics. Methods:The orthogonal experimental design was used to screen out the optimal formulation and process of exenatide-loaded PLGA nanoparticles,and dynamic light scattering and a transmission electron microscope were used for characterization. In vitro simulation of gastrointestinal fluid and Caco-2 cell model were used to evaluate its stability,cellular uptake,and cellular transport. Results:The optimal formulation and process were obtained by the orthogonal design,i.e.,a volume ratio of internal water phase to oil phase of 1∶5,an ultrasonic power of 200 W,an ultrasonic time of 4 minutes,a concentration of polyvinyl alcohol of2%,and a volume ratio of primary emulsion to external water phase of 1∶20. The nanoparticles obtained had a diameter of 116.2 nm and the transmission electron microscope showed that PLGA nanoparticles had a sphere-like shape with a regular surface. The in vitro study demonstrated that PLGA nanoparticles significantly prolonged the release of exenatide in gastrointestinal fluid and improved its stability(P<0.05),and 50% of exenatide was retained after 4-hour culture in simulated gastric fluid,while 80% was retained after4-hour culture in simulated intestinal juice. The cell experiment showed that the uptake and transport of PLGA nanoparticles in the experimental group were 7 and 5 times those in the control group(P<0.05). Conclusion:PLGA nanoparticles may be a potential efficient oral drug delivery system for exenatide.
Objective:To prepare exenatide-loaded poly(lactic-co-glycolic acid)(PLGA)nanoparticles,and to evaluate its in vitro effect and characteristics. Methods:The orthogonal experimental design was used to screen out the optimal formulation and process of exenatide-loaded PLGA nanoparticles,and dynamic light scattering and a transmission electron microscope were used for characterization. In vitro simulation of gastrointestinal fluid and Caco-2 cell model were used to evaluate its stability,cellular uptake,and cellular transport. Results:The optimal formulation and process were obtained by the orthogonal design,i.e.,a volume ratio of internal water phase to oil phase of 1∶5,an ultrasonic power of 200 W,an ultrasonic time of 4 minutes,a concentration of polyvinyl alcohol of2%,and a volume ratio of primary emulsion to external water phase of 1∶20. The nanoparticles obtained had a diameter of 116.2 nm and the transmission electron microscope showed that PLGA nanoparticles had a sphere-like shape with a regular surface. The in vitro study demonstrated that PLGA nanoparticles significantly prolonged the release of exenatide in gastrointestinal fluid and improved its stability(P<0.05),and 50% of exenatide was retained after 4-hour culture in simulated gastric fluid,while 80% was retained after4-hour culture in simulated intestinal juice. The cell experiment showed that the uptake and transport of PLGA nanoparticles in the experimental group were 7 and 5 times those in the control group(P<0.05). Conclusion:PLGA nanoparticles may be a potential efficient oral drug delivery system for exenatide.
引文
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[13]谢鑫鑫,王俊腾,奉利芳,等.聚乙二醇化聚乳酸羟基乙酸纳米粒的制备及体外抗粘性能评价[J].中国医院药学杂志,2012,32(21):1727-1231.
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[16] Silva AL,Rosalia RA,Sazak A,et al. Optimization of encapsulation of a synthetic long peptide in PLGA nanoparticles:low-burst release is crucial for efficient CD8+T cell activation[J]. Eur J Pharm Biopharm,2013,83(3):338-345.
[17] Joshi G,Kumar A,Sawant K. Bioavailability enhancement,Caco-2 cells uptake and intestinal transport of orally administered lopinavir loaded PLGA nanoparticles[J]. Drug Deliv,2016,23(9):3492-3504.
[2]於丽红,李春慧,苏胜偶,等.不同时间注射甘精胰岛素治疗1型糖尿病的安全性及疗效评价[J].河北医科大学学报,2014,35(4):395-399.
[3] Meier JJ. GLP-1 receptor agonists for individualized treatment of type 2 diabetes mellitus[J]. Nature Rev Endocrinol,2012,8(12):728-742.
[4] Nauck MA,Vardarli I,Deacon CF,et al. Secretion of glucagon-like peptide-1(GLP-1)in type 2 diabetes:what is up,what is down?[J].Diabetologia,2011,54(1):10-18.
[5]张文静,付阿丹,赵湜.艾塞那肽治疗2型糖尿病患者的疗效及安全性评价[J].中国医院药学杂志,2012,32(9):723-725.
[6] Soudry-Kochavi L,Naraykin N,Nassar T,et al. Improved oral absorption of exenatide using an original nanoencapsulation and microencapsulation approach[J]. J Control Release,2015,217:202-210.
[7] Gupta S,Jain A,Chakraborty M,et al. Oral delivery of therapeutic proteins and peptides:a review on recent developments[J]. Drug Deliv,2013,20(6):237-246.
[8] Malathi S,Nandhakumar P,Pandiyan V,et al. Novel PLGA-based nanoparticles for the oral delivery of insulin[J]. Int J Nanomedicine,2015,10:2207-2218.
[9] Jain S,Rathi VV,Jain AK,et al. Folate-decorated PLGA nanoparticles as a rationally designed vehicle for the oral delivery of insulin[J].Nanomedicine(Lond),2016,7(9):1311-1337.
[10] Zhang X,Sun M,Zheng A,et al. Preparation and characterization of insulin-loaded bioadhesive PLGA nanoparticles for oral administration[J]. Eur J Pharm Sci,2012,45(5):632-638.
[11]宋逸婷,沈剑锋,钱帅,等. PLGA黏度及PVA浓度对微球形成过程中物理性质的影响[J].中国医药工业杂志,2010,41(4):269-272.
[12]樊莉,鲁莹,吴诚,等.干扰素α-2b复合物PLGA微球的制备及影响因素[J].中国医药工业杂志,2006,37(10):674-678.
[13]谢鑫鑫,王俊腾,奉利芳,等.聚乙二醇化聚乳酸羟基乙酸纳米粒的制备及体外抗粘性能评价[J].中国医院药学杂志,2012,32(21):1727-1231.
[14] Cui M,Wu W,Hovgaard L,et al. Liposomes containing cholesterol analogues of botanical origin as drug delivery systems to enhance the oral absorption of insulin[J]. Int J Pharm,2015,489(1-2):277-284.
[15] Fornaguera C,Dols-Perez A,CalderóG,et al. PLGA nanoparticles prepared by nano-emulsion templating using low-energy methods as efficient nanocarriers for drug delivery across the blood-brain barrier[J]. J Control Release,2015,211:134-143.
[16] Silva AL,Rosalia RA,Sazak A,et al. Optimization of encapsulation of a synthetic long peptide in PLGA nanoparticles:low-burst release is crucial for efficient CD8+T cell activation[J]. Eur J Pharm Biopharm,2013,83(3):338-345.
[17] Joshi G,Kumar A,Sawant K. Bioavailability enhancement,Caco-2 cells uptake and intestinal transport of orally administered lopinavir loaded PLGA nanoparticles[J]. Drug Deliv,2016,23(9):3492-3504.