过氧化氢敏感的靶向荧光载药纳米粒子的制备及在动脉粥样硬化中的应用

doi:10.7503/cjcu20180033

摘要/Abstract

摘要：

活性氧(ROS)在动脉粥样硬化斑块的形成过程中起着重要作用, 特别是过氧化氢在动脉硬化内皮损伤处有着较高含量, 其可以刺激巨噬细胞表达促炎细胞因子和趋化因子来加剧巨噬细胞的致炎活动. 因此, 开发具有过氧化氢敏感释放的药物, 对于安全有效治疗动脉粥样硬化具有重大意义. 将聚甲基丙烯酸羟乙酯(PHEMA)侧链羟基分别接枝草酰基辛伐他汀(Sim)(降低血脂前药)、含氧杂六元环的荧光聚乙二醇(FPEG)及巨噬细胞的靶分子ISO-1, 制备了具有过氧化氢敏感型的靶向荧光载药纳米胶束(PHEMA-Sim-FPEG-ISO-1). 该纳米粒子利用过氧化氢敏感可控释药, 能够靶向巨噬细胞, 同时改进传统聚乙二醇仅作亲水链的特点, 合成具有荧光功能的聚乙二醇, 使聚合物无需修饰荧光物质便拥有探针能力, 适于动脉粥样硬化的诊疗. 结果表明, 该水溶性纳米粒子显示出较好的生物相容性、过氧化氢敏感性及荧光能力, 为发展动脉粥样硬化的诊疗一体的纳米体系提供新材料.

关键词: 两亲纳米粒子, 过氧化氢敏感, 药物释放, 荧光聚合物, 动脉粥样硬化

Abstract:

A multifunctional nanomicelle was designed for atherosclerosis theranostics, which was synthesized by decorating the poly(2-hydroxyethyl methacrylate)(PHEMA) with simvastatin(Sim) prodrug, fluorescent polyethylene glycol(FPEG) and the macrophages targeting molecules(ISO-1). The as-obtained polymer was confirmed by nuclear magnetic resonance(¹H NMR), Fourier transform infrared spectroscopy(FTIR) and gel permeation chromatography(GPC). The nanoparticle size and morphology were determined by dynamic light scattering(DLS) and transmission electron microscopy(TEM), which was spherical and with the size of about 100 nm. The MTT assay confirmed the biocompatibility of the nanomicelles. The targeting property was evaluated by cell uptake assay and observed via confocal laser scanning microscopy(CLSM). The results suggest that this multifunctional H₂O₂-respective targeting nanomicelles could become an effective tool for atherosclerosis therapy.

Key words: Amphiphilic nanoparticles, Hydrogen peroxide sensitivity, Drug release, Fluorescent polymer, Atherosclerosis

中图分类号:

O631

TrendMD:

方超, 朱焓毓, 刘晔, 赵外欧, 李亚鹏, 王静媛. 过氧化氢敏感的靶向荧光载药纳米粒子的制备及在动脉粥样硬化中的应用. 高等学校化学学报, 2018, 39(9): 2071.

FANG Chao,ZHU Hanyu,LIU Ye,ZHAO Waiou,LI Yapeng,WANG Jingyuan. Synthesis and Characterization of Nanoparticles with Hydrogen Peroxide Sensitivity, Targeting and Fluorscence for Atherosclerosis. Chem. J. Chinese Universities, 2018, 39(9): 2071.

图/表 10

Scheme 1 Synthesis of multifunctional polymer PHEMA-Sim-FPEG-ISO-1

Scheme 2 Synthesis routes of PSFI

Fig.1 1H NMR spectrum(A), FTIR spectra(B) and GPC curves(C) of PSFI Peaks 1—8 are corresponding to the positions 1—8 in Scheme 2. a. PHEMA; b. PHEMA-Sim; c. PSFI.

Table 1 Characteristics of PHEMA-Sim-FPEG

n(Sim)-n(FPEG)	10⁴ cmc/(mg·mL^-1)	Size/nm	PDI	Yield/g	Yield rate(%)
1-1	77.60	225.5±2.4	0.189±0.035	0.844	53.4
2-1	63.30	211.2±5.6	0.221±0.022	0.468	45.9
3-1	25.20	192.6±3.8	0.205±0.019	0.397	47.6
4-1	8.69	154.3±4.2	0.211±0.017	0.324	43.8
5-1	8.90	165.5±1.0	0.169±0.035	0.268	39.2

Fig.2 TEM image(A) and DLS(B) of PSFI

Fig.3 Fluorescence excitation spectra(A) and emission spectra(B) of FPEG in PBS(a), PSFI in PBS(b) and PSFI in 1 mmol/L H2O2(c)

Fig.4 Drug release curves of PSFI in H2O2c(H2O2)/(mmol·L-1): a. 0; b. 0.25; c. 0.50; d. 0.75; e. 1.

Fig.5 Cytotoxicity evaluation of PSFI in macrophages RAW264.7(A) and endothelial cells RBMEC(B) and PHEMA-Sim-FPEG, PHEMA-Sim-RhoB, PHEMA-Sim-5-FAM, PHEMA-Sim-PEG@QDs in macrophages RAW264.7 24 h(C)

Fig.6 Confocal laser scanning microscope images of blank(A1—A4), PSF(B1—B4) and PSFI(C1—C4) in macrophages RAW264.7(A1—C1) F-actin; (A2—C2) nuclear; (A3—C3) nanoparticles; (A4—C4) merge.

Fig.7 Confolcal laser scanning microscope images of blank(A1—A4), PSF(B1—B4) and PSFI(C1—C4) in endothelial cells RBMEC in mice(A1—C1) F-actin; (A2—C2) nuclear; (A3—C3) nanoparticles; (A4—C4) merge.

参考文献 42

[1]	Mark E. L., Claudia C., Antoine M., Max L. S., Francois F., Philip M. R., Sarayu R., Tina B., Maarten P., Steven S., Stephan R., Ronald E. G., Luis C. Z., Gert S., Josbert M. M., Christopher H. C., Erik S. G., Zahi A. F., Willem J. M., ACS Nano, 2015, 9(2), 1837—1847
[2]	Tuttolomondo A., Di R. D., Pecoraro R., Arnao V., Pinto A., Licata G., Curr. Pharm. Design, 2012, 18(28), 4266—4288
[3]	Hadeel A. Y., J. Fac. Med. Baghdad, 2014, 56(2), 186—190
[4]	Yamashita T., Sasaki N., Kasahara K., Hirata K. I., J. Cardiol., 2015, 66(1), 1—8
[5]	Schiener M., Hossann M., Viola J. R., Ortega-Gomez A., Weber C., Lauber K., Lindner L. H., Soehnlein O., Trends Mol. Med., 2014, 20(5), 71—81
[6]	Wong B. W., Meredith A., Lin D., Mcmanus B. M., Can. J. Cardiol., 2012, 28(6), 631—641
[7]	Petros R. A., De Simone J. M., Nat. Rev. Drug Discov., 2015, 9, 615—627
[8]	Brannon P. L., Blanchette J. O., Adv. Drug Deliver Rev., 2004, 56(11), 1649—1659
[9]	Tapeinos C., Pandit A., Adv. Mater., 2016, 28(27), 5553—5585
[10]	Madamanchi N. R., Vendrov A., Runge M. S., Arterioscl. Throm. Vas., 2005, 25(1), 29—38
[11]	Tong R., Tang L., Ma L., Tu C., Baumgartner R., Cheng J., Chem. Soc. Rev., 2014, 43(20), 6982—7012
[12]	Zhang Y., Yu J., Bomba H. N., Zhu Y., Gu Z., Chem. Rev., 2016, 116(19), 12536—12563
[13]	Sun X. C., Yang H., Wang J. Z., Ma R. J., An Y. L., Shi L. Q., Chem. J. Chinese Universities, 2014, 35(7), 1570—1578
	(孙小成, 杨浩, 王建祖, 马如江, 安英丽, 史林启. 高等学校化学学报, 2014, 35(7), 1570—1578)
[14]	Lee S. H., Gupta M. K., Bang J. B., Bae H., Sung H. J., Adv. Healthcare Mater., 2013, 2(6), 908—915
[15]	Yan Q., Yuan J. Y., Chem. J. Chinese Universities, 2012, 33(9), 1877—1885
	(闫强, 袁金颖. 高等学校化学学报, 2012, 33(9), 1877—1885)
[16]	Liu X., Xiang J. J., Zhu D. C., Jiang L. M., Zhou Z. X., Tang J. B., Liu X. R., Huang Y. Z., Shen Y. Q., Adv. Mater., 2016, 28(9), 1743—1752
[17]	Xu Q. H., He C. L., Xiao C. S., Chen X. S., Macromol. Biosci., 2016, 16(5), 635—646
[18]	Christos T., Abhay P., Adv. Mater., 2016, 28, 5553—5585
[19]	Marian V., Dieter L., Jan M., Mark T. D., Milan M., Joshua T., Int. J. Biochem. & Cell Biology, 2007, 39, 44—84
[20]	Barnes T. C., Anderson M. E., Edwards S. W., Moots R. J., Rheumatology, 2012, 51, 1166—1169
[21]	Manuela C., Ileana M., Curr. Med. Chem., 2017, 24, 550—567
[22]	Velasquez J. C., Weiss D., Joseph G., Landazuri N., Taylor W. R., Circulation, 2008, 118(18), S510
[23]	Wang Y. L., Sun G. Y., Zhang Y., He J. J., Zheng S., Lin J. N., Mol. Med. Rep., 2016, 14, 3559—3564
[24]	Guanying C., Indrajit R., Chunhui Y., Paras N.P., Chem. Rev., 2016, 116, 2826—2885
[25]	Mahmoud E., Gyu S., Soon-Mi L., Guorong S., Karen L.W., Chem. Rev., 2015, 115, 10967—11011
[26]	Karel U., Kateǐina H., Vladimir S., Aristides B., Jiǐí T., Radek Z., Chem. Rev., 2016, 116, 5338—5431
[27]	Calandra T., Roger T., Nat. Rev. Immunol., 2003, 3(10), 791—800
[28]	Chang D., Wang Y. C., Zhang S. J., Bai Y. Y., Liu D. F., Zang F. C., Wang G. Z., Wang B. H., Ju S. H., Biomaterials, 2015, 68, 67—76
[29]	Gui R. J., An X. Q., Huang W. X., Anal. Chim. Acta, 2013, 767, 134—140
[30]	Gyawali D., Zhou S. Y., Tran R. T., Zhang Y., Liu C., Bai X. C., Yan. J., Adv. Healthcare Mater., 2014, 3(2), 182—186
[31]	Yang J., Zhang Y., Gautam S., Liu L., Dey J., Chen W., Mason R. P., Serrano C. A., Schug K. A., Tang L. P., Natl. Acad. Sci. USA, 2009, 106(25), 10086—10091
[32]	Cheng K. F., Al-Abed Y., Bioorg. Med. Chem. Lett., 2006, 16(13), 3376—3379
[33]	Gao H., Matyjaszewski K., J. Am. Chem. Soc., 2007, 129(20), 6633—6639
[34]	Wang J., Greszta DMatyjaszewski K., Polym. Mater. Sci. Eng., 1995, 73, 416—417
[35]	Jiao Z., Liu N., Chen Z. M., Pharm. Dev. Technol., 2012, 17, 164—169
[36]	Lee D., Khaja S., Velasquezcastano J. C., Madhuri D., Carrie S., John P. W., Taylor R., Niren M., Nat. Mater., 2007, 6(10), 765—769
[37]	Carsten S., Stephan L. M., Martin P., Pieter A., Clin. Exp. Pharmacol. P, 2003, 30, 249—253
[38]	Burak C. S., Ayse S. Ş., Exp. Therap. Med., 2014, 8, 1660—1664
[39]	Calin M., Manduteanu I., Curr. Med. Chem., 2017, 24, 550—567
[40]	Brown B. G., Zhao Q. X., Chait A., N. Engl. J. Med., 2001, 345(22), 1583—1592
[41]	Pavia C. S., Folds J. D., Baseman J. B., Infect. Immun., 1977, 17(3), 651—654
[42]	Sarala B., Atish R., Pradip K. G., Vitthal N. Y., Divya K., Anshu C., Pooja B., Shaila S., Somesh S., Ram A. V., Nilesh M. D., Bioorg. Med. Chem. Lett., 2005, 19(16), 4773—4776