Preparation and identification of epidermal growth factor coupling bovine serum albumin nanoparticles loading paclitaxel

doi:10.3969/j.issn.2095-4344.2014.52.006

Abstract

Abstract:

BACKGROUND: Paclitaxel is a broad-spectrum antitumor plant drug commonly used, possessing much adverse reactions. Therefore, there is an urgent need for an appropriate carrier to reduce the toxicity of paclitaxel, and achieve better targeting.

OBJECTIVE: To explore the preparation of epidermal growth factor (EGF) coupling bovine serum albumin nanoparticles loading paclitaxel and its physical and chemical properties.

METHODS: Albumin nanoparticles were prepared by phacoemulsification and solvent evaporation technology, and EGF was coupled with albumin nanoparticles by chemical crosslinking reagents. The EGF coupling albumin nanoparticles loading ¹²⁵I marked paclitaxel acted as experimental samples. 100, 200, 400, 800 mg/L ¹²⁵I marked paclitaxel was added to albumin nanoparticles and EGF coupling albumin nanoparticles respectively for detection of encapsulation efficiency and loading rate. The drug releasing rate of samples and albumin nanoparticles loading ¹²⁵I marked paclitaxel were measured. The stability at room temperature and serum stability of samples, albumin nanoparticles loading ¹²⁵I marked paclitaxel and ¹²⁵I marked paclitaxel were detected.

RESULTS AND CONCLUSION: With the increasing of paclitaxel dosage, in albumin nanoparticles loading ¹²⁵I marked paclitaxel group and experimental sample group, the encapsulation efficiency showed a trend of decline but the drug loading rate presented a rising tendency. Experimental sample group and albumin nanoparticles loading¹²⁵I marked paclitaxel group showed the same releasing drug trend basically, manifesting the sustained release of paclitaxel. The stability at room temperature and serum stability of experimental sample group and albumin nanoparticles loading ¹²⁵I marked paclitaxel group were both higher than those in¹²⁵I marked paclitaxel group (P < 0.05). From what has been discussed above, EGF coupling albumin nanoparticles loading ¹²⁵I marked paclitaxel have good drug-loading rate, drug release rate and stability.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

全文链接：

Key words: epidermal growth factor, serum albumin, bovine, nanoparticles

CLC Number:

R318

Wang Shu-bin, Yuan Fei, Peng Zhi-ping, Li Shao-lin, Wu Yun, Zhu Wei. Preparation and identification of epidermal growth factor coupling bovine serum albumin nanoparticles loading paclitaxel

[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(52): 8393-8398.

Figures/Tables 1

2.1 荷载125I标记紫杉醇表皮生长因子-牛血清白蛋白纳米微粒的载药量和包封率表皮生长因子-牛血清白蛋白纳米微粒对125I标记紫杉醇的包载及其载药量和包封率的测量具体数据见表1，可见荷载125I标记紫杉醇牛血清白蛋白纳米微粒和荷载125I标记紫杉醇表皮生长因子-牛血清白蛋白纳米微粒的包封率和载药量随着紫杉醇药量的增加，包封率呈现下降的趋势，载药量呈现上升的趋势，分别进行配对t 检验，差异无显著性意义(P > 0.05)。 2.2 荷载125I标记紫杉醇表皮生长因子-牛血清白蛋白纳米微粒体外药物释放的测定荷载125I标记紫杉醇表皮生长因子-牛血清白蛋白纳米微粒的体外释药情况如表2所示，从中可以看出，荷载125I标记紫杉醇牛血清白蛋白纳米微粒和荷载125I标记紫杉醇表皮生长因子-牛血清白蛋白纳米微粒的趋势基本相同，表现为包载药物缓慢释放，分别进行配对t 检验，差异无显著性意义(P > 0.05)。 2.3 荷载125I标记紫杉醇的表皮生长因子-牛血清白蛋白纳米微粒的稳定性 2.3.1 室温稳定性在室温放置1，6，12，24 h进行TLC，见表3。1 h时荷载125I标记紫杉醇的牛血清白蛋白纳米微粒和荷载125I标记紫杉醇的表皮生长因子-牛血清白蛋白纳米微粒测得的化学纯> 95%，6 h时略有降低(> 85%)，但差异无显著意义(P > 0.05)。采用统计软件统计分析得：同一时间点组内差异无显著性意义(P > 0.05)，在24 h内放化纯度比较差异有显著性意义(P < 0.05)，做回归分析，F 值为36.75时间与放射纯度呈线性相关，回归方程为Y=96.34-3.275X，R2为0.957(n=3)。结果显示，紫杉醇被靶向纳米粒包载后在中性pH溶液中，室温24 h内具有较好的稳定性，125I不易脱落，较未用靶向纳米粒包载的125I标记紫杉醇室温稳定性好。 2.3.2 血清稳定性荷载125I标记紫杉醇的表皮生长因子-牛血清白蛋白纳米微粒在新鲜人血清中37 ℃孵育0.5，1，2，4 h，用体积分数75%乙醇展开剂纸层析法测定放射化学纯度，结果见表4。4 h放射化学纯度略有降低，与0.5 h相比差异无显著性意义(P > 0.05)。做回归分析F 值为88.37，时间与放射纯度呈线性相关，回归方程为Y=96.23-3.874X，R2为0.964(n=3)。结果显示，荷载125I标记紫杉醇的表皮生长因子-牛血清白蛋白纳米微粒在血清中24 h内具有较好的稳定性，125I不易脱落。荷载125I标记紫杉醇的表皮生长因子-牛血清白蛋白纳米微粒、荷载125I标记紫杉醇的牛血清白蛋白纳米微粒在各个时间点的细胞摄取率都明显高于125I标记紫杉醇组。通过方差分析，各药物组间均方MS=10.285，F=4.654，R2=0.965，P < 0.01。3组的血清稳定性有统计学差异，荷载125I标记紫杉醇的表皮生长因子-牛血清白蛋白纳米微粒更为稳定，能够满足体内分析。 "

References

[1] 赵平.中国癌症流行态势与对策[J].医学研究杂志, 2013,42(10):3.
[2] Digumartia R,Bapsy PP,Suresh A,et al.Bavituximab plus paclitaxel and carboplatin for the treatment of advanced non-small-cell lung cancer. Lung Cancer.2014;86(2):231-236.
[3] Lam SW,de Groot SM,Honkoop AH,et al.Paclitaxel and bevacizumab with or without capecitabine as first-line treatment for HER2-negative locally recurrent or metastatic breast cancer: A multicentre, open-label, randomised phase 2 trial. Eur J Cancer.2014.
[4] Polee MB,J Verweij J,Siersema PD,et al.Phase I study of a weekly schedule of a fixed dose of cisplatin and escalating doses of paclitaxel in patients with advanced oesophageal cancer. Eur J Cancer.2002;38(11):1495-1500.
[5] van der Burg MEL,Onstenk W,Boere IA,et al. Long-term results of a randomised phase III trial of weekly versus three-weekly paclitaxel/platinum induction therapy followed by standard or extended three-weekly paclitaxel/platinum in European patients with advanced epithelial ovarian cancer. Eur J Cancer.2014;50(15):2592-2601.
[6] Lorusso D,Petrelli F,Coinu A,et al. A systematic review comparing cisplatin and carboplatin plus paclitaxel-based chemotherapy for recurrent or metastatic cervical cancer. Gynecol Oncol.2014;133(1):117-123.
[7] Huang D,Ba Y,Xiong J,et al.A multicentre randomised trial comparing weekly paclitaxel + S-1 with weekly paclitaxel + 5-fluorouracil for patients with advanced gastric cancer. Eur J Cancer.2013;49(14):2995-3002.
[8] Kim HY,Ryu JH,Chu CW,et al.Paclitaxel-incorporated nanoparticles using block copolymers composed of poly(ethylene glycol)/poly(3-hydroxyoctanoate). Nanoscale Res Lett.2014;9(1):525.
[9] Rowinsky EK,Donehower RC.Paclitaxel (taxol).N Engl J Med.1995;332(15):1004-1014.
[10] Kobayashi Y,Seino K,Hosonuma S,et al.Side population is increased in paclitaxel-resistant ovarian cancer cell lines regardless of resistance to cisplatin. Gynecol Oncol.2011; 121(2):390-394.
[11] Kamath KR,Barry JJ,Miller KM.The Taxus™ drug-eluting stent: a new paradigm in controlled drug delivery.Adv Drug Deliv Rev.2006;58(3):412-436.
[12] 王树斌,袁飞,武云,等.纳米载体在肿瘤靶向治疗中的应用[J].中国组织工程研究与临床康复,2009,13(29):5739-5742.
[13] Endres NF,Barros T,Cantor AJ,et al.Emerging concepts in the regulation of the EGF receptor and other receptor tyrosine kinases.Trends Biochem Sci.2014;39(10):437-446.
[14] Cheng X,Chen H.Tumor heterogeneity and resistance to EGFR-targeted therapy in advanced nonsmall cell lung cancer: challenges and perspectives. Onco Targets Ther. 2014;7:1689-1704.
[15] 王树斌,袁飞,彭志平,等.EGF偶联白蛋白靶向纳米载体在荷瘤裸鼠体内分布的研究[J].中国肿瘤临床,2008,35(9):516-519.
[16] Coolbrandt A,Van den Heede K,Vanhove E,et al.Immediate versus delayed self-reporting of symptoms and side effects during chemotherapy:Does timing matter? Eur J Oncol Nurs. 2011;15(2):130-136.
[17] Acharya G,Park K.Mechanisms of controlled drug release from drug-eluting stents.Adv Drug Deliv Rev.2006;58(3):387-401.
[18] Liu SJ,Hsiao CY,Chen JK,et al.In-vitro release of anti-proliferative paclitaxel from novel balloon-expandable polycaprolactone stents.Mater Sci Eng.2011;31:1129-1135.
[19] Joerger M.Metabolism of the taxanes including nab-paclitaxel. Expert Opin Drug Metab Toxicol.2014;14:1-12.
[20] Vrignaud P, Semiond D, Benning V, et al.Preclinical profile of cabazitaxel. Drug Des Devel Ther. 2014;8:1851-1867.
[21] Wang JJ,Huang SW.Research Progress on Novel Carrier-modified Methods and Evaluation of Active Targeting Antitumor Preparation. Chin Herbal Med.2014;6(1):22-28.
[22] Costa A,Sarmento B,Seabra V.Targeted Drug Delivery Systems for Lung Macrophages.Curr Drug Targets.2014. [Epub ahead of print]
[23] Horváti K, Bacsa B, Kiss E, et al. Nanoparticle encapsulated lipopeptide conjugate of antitubercular drug isoniazid: in vitro intracellular activity and in vivo efficacy in a guinea pig model of tuberculosis.Bioconjug Chem.2014.[Epub ahead of print]
[24] Scalia S,Young PM,Traini D.Solid lipid microparticles as an approach to drug delivery.Expert Opin Drug Deliv.2014;13:1-17.
[25] Wei Y,Li L,Xi Y,et al.Sustained release and enhanced bioavailability of injectable scutellarin-loaded bovine serum albuminnanoparticles.Int J Pharm.2014;476(1-2):142-148.
[26] Nitta SK,Numata K.Biopolymer-based nanoparticles for drug/gene delivery and tissue engineering.Int J Mol Sci.2013; 14(1):1629-1654.
[27] Liu TY,Chen SY,Liu DM,et al.On the study of BSA-loaded calcium-deficient hydroxyapatite nano-carriers for controlled drug delivery. J Control Release.2005;107(1):112-121.
[28] Daraee H,Eatemadi A,Abbasi E,et al.Application of gold nanoparticles in biomedical and drug delivery.Artif Cells Nanomed Biotechnol.2014;17:1-13.
[29] Birnbaum DT,Kosmala JD,Henthorn DB,et al.Controlled release of estradiol from PLGA microparticles:the effect of organic phase solvent on encapsulation and realease.J Controll Release.2000;65(3):375.
[30] Mane SR,Dinda H,Sathyan A,et al.Increased bioavailability of rifampicin from stimuli-responsive smart nano carrier.ACS Appl Mater Interfaces. 2014;6(19):16895-1902.
[31] Huang D,Wang L,Dong Y,et al.A novel technology using transscleral ultrasound to deliver protein loaded nanoparticles.Eur J Pharm Biopharm.2014;88(1):104-115.
[32] Barua S,Mitragotri S.Challenges associated with Penetration of Nanoparticles across Cell and Tissue Barriers: A Review of Current Status and Future Prospects. Nano Today.2014;9(2): 223-243.
[33] Raj S,Jose S,Sumod US,et al.Nanotechnology in cosmetics: Opportunities and challenges. J Pharm Bioallied Sci.2012; 4(3):186-193.
[34] Mooso BA,Vinall RL,Mudryj M,et al.The Role of EGFR Family Inhibitors in Muscle Invasive Bladder Cancer: A Review of Clinical Data and Molecular Evidence. J Urol. 2014. pii: S0022-5347(14)04269-4.
[35] Ou SH.Second-generation irreversible epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs): a better mousetrap? A review of the clinical evidence.Crit Rev Oncol Hematol.2012;83(3):407-421.
[36] Cadranel J, Ruppert AM, Beau-Faller M,et al.Therapeutic strategy for advanced EGFR mutant non-small-cell lung carcinoma.Crit Rev Oncol Hematol.2013;88(3):477-493.
[37] Brand TM,Iida M,Luthar N,et al.Nuclear EGFR as a molecular target in cancer. Radiother Oncol.2013;108(3):370-377.
[38] Chaturvedi SK,Ahmad E,Khan JM,et al.Elucidating the interaction of limonene with bovine serum albumin: a multi-technique approach. Mol Biosyst. 2014.[Epub ahead of print]
[39] Shi Y,Su C,Cui W,et al. Gefitinib loaded folate decorated bovine serum albumin conjugated carboxymethyl-beta- cyclodextrin nanoparticles enhance drug delivery and attenuate autophagy in folate receptor-positive cancer cells.J Nanobiotechnology. 2014;12(1):43.