pH非敏感型自交联共轭亚油酸囊泡的构建和表征

doi:10.7503/cjcu20140158

摘要/Abstract

摘要：

以共轭亚油酸(Conjugated linoleic acid, CLA)为构造pH非敏感型脂肪酸囊泡(Fatty acid vesicle, FAV)的分子砌块, 通过碱异构化法从亚油酸半合成CLA, 然后采用pH刺激响应自组装法获得CLA的不饱和脂肪酸囊泡(Ufasome), 采用紫外辐照诱导方式对CLA-ufasome实施囊泡内化学绑定, 获得一种新的pH非敏感型FAV. 通过酸碱滴定和表面张力实验确定CLA-ufasome形成的适宜pH范围和浓度, 利用透射电子显微镜(TEM)表征了自交联CLA的FAV的形貌, 并通过动态光散射(DLS)法测定了自交联CLA的FAV的稳定性. 结果表明, 以CLA为分子砌块, 当浓度为3 mmol/L时在pH=8.6条件下构建CLA-ufasome, 紫外辐照2.5 h后得到粒径为10~20 nm, 壁厚为2.0 nm的自交联CLA的FAV, 并具有pH非敏感的特性. 以抗癌药物五氟尿嘧啶为目标包覆药物, 体外释放实验结果表明, 自交联CLA的FAV对五氟尿嘧啶具有良好的缓释效果.

关键词: 共轭亚油酸, 不饱和酸囊泡, 脂肪酸囊泡, pH非敏感性, 自组装, 自交联

Abstract:

Conjugated linoleic acid(CLA) was employed as a molecular building block to construct a novel pH-insensitive fatty acid vesicle(FAV). First, CLA was semi-synthesized from linoleic acid through alkali isomerization. Subsequently, the ufasome(unsaturated fatty acid liposome) of CLA was self-assembled from the CLA molecules together with the CLA ions in response to pH, and a novel pH-insensitive FAV, the self-crosslinked-CLA FAV, was prepared by chemical tethering the CLA-ufasome through UV irradiation. The sui-table pH ranges and the CLA concentrations were determined by acid-base titration and surface tension mea-surement. The morphology of the self-crosslinked-CLA FAV was imaged by transmission electro microscopy(TEM) and the pH-insensitive characteristic of the self-crosslinked-CLA FAV was tested by dynamic light scattering(DLS). The experimental results indicated that the self-crosslinked-CLA FAV with diameter of 10—20 nm and wall thickness of 2.0 nm was obtained through a two-step procedure. At first, the CLA-ufasome formed using the CLA species as a molecular building block at pH=8.6 and CLA concentration of 3 mmol/L, and then followed by UV irradiation self-crosslinking for 2.5 h. It was verified by DLS data that the self-crosslinked-CLA FAV is pH-insensitive in almost all pH ranges. Finally, the in vitro release results of 5-fluorouracil from the self-crosslinked-CLA FAV showed that the process was slow and sustainable.

Key words: Conjugated linoleic acid, Ufasome, Fatty acid vesicle, pH-Insensitive, Self-assembling, Self-crosslinking

中图分类号:

O648

TrendMD:

樊晔, 方云, 高迪. pH非敏感型自交联共轭亚油酸囊泡的构建和表征. 高等学校化学学报, 2014, 35(8): 1746.

FAN Ye, FANG Yun, GAO Di. Construction and Characterization of a Novel pH-insensitive Self-crosslinked Ufasome of Conjugated Linoleic Acid^†. Chem. J. Chinese Universities, 2014, 35(8): 1746.

图/表 6

Fig.1 Titration curve of 10 mmol/L CLA at 25 ℃ Area a. micelle; b. ufasome; c. emulsion; d. oil drop.

Fig.2 γ-lgc curve of CLA with pH=8.6 at 25 ℃

Fig.3 UV spectra(A) and IR spectra(B) of the aqueous CLA solution at pH=8.6 before and after UV irradiation for 2.5 h and the size distribution(C) and TEM image(D) of the 3 mmol/L self-crosslinked-CLA FAV using QTX as initiator

Fig.4 Schematic illustration of the formation of the self-crosslinked-CLA FAV

Fig.5 pH insensitive test of the self-crosslinked-CLA FAV (A) Before UV irradiation; (B)—(D) after UV irradiation; (C), (D) DLS and corresponding diameter distribution with different pH values, respectively.

Fig.6 In vitro released rate of 5-FU from the self-crosslinked-CLA FAV in PBS buffer

参考文献 41

[1]	Gebicki J., Hicks M., Nature, 1973, 243(5404), 232—234
[2]	Hargreaves W. R., Deamer D. W., Biochem., 1978, 17(18), 3759—3768
[3]	Cistola D. P., Hamilton J. A., Jackson D., Small D. M., Biochem., 1988, 27(6), 1881—1888
[4]	Morigaki K., Walde P., Misran M., Robinson B. H., Colloids Surf. A, 2003, 213(1), 37—44
[5]	Gebicki J. M., Hicks M., Chem. Phys. Lipids, 1976, 16(2), 142—160
[6]	Namani T., Ishikawa T., Morigaki K., Walde P., Colloids Surf. B, 2007, 54(1), 118—123
[7]	Namani T., Walde P., Langmuir, 2005, 21(14), 6210—6219
[8]	Rogerson M. L., Robinson B. H., Bucak S., Walde P., Colloids Surf. B, 2006, 48(1), 24—34
[9]	Chen I. A., Salehi-Ashtiani K., Szostak J. W., J. Am. Chem. Soc., 2005, 127(38), 13213—13219
[10]	Dowling M. B., Lee J. H., Raghavan S. R., Langmuir, 2009, 25(15), 8519—8525
[11]	Markvoort A. J., Pfleger N., Staffhorst R., Hilbers P. A. J., van Santen R. A., Killian J. A., de Kruijff B., Biophys. J., 2010, 99(5), 1520—1528
[12]	Xu W., Wang X., Zhong Z., Song A., Hao J., J. Phys. Chem. B, 2013, 117(1), 242—251
[13]	Morigaki K., Walde P., Curr. Opin. Colloid Interface Sci., 2007, 12(2), 75—80
[14]	Deamer D., Singaram S., Rajamani S., Kompanichenko V., Guggenheim S., Philos. T. Roy. Soc. B, 2006, 361(1474), 1809—1818
[15]	Fukuda H., Goto A., Yoshioka H., Goto R., Morigaki K., Walde P., Langmuir, 2001, 17(14), 4223—4231
[16]	Li X., Ni X. F., Liang Z. H., Shen Z. Q., Chem. J. Chinese Universities, 2012, 33(6), 1333—1338
	(李雪, 倪旭峰, 梁振华, 沈之荃. 高等学校化学学报, 2012, 33(6), 1333—1338)
[17]	Liu Y. H., Li G. J., Luo X. W., Peng X. Y., Wang L. Y., Chem. J. Chinese Universities, 2013, 34(6), 1527—1535
	(刘云鸿, 李光吉, 罗熙雯, 彭新艳, 王立莹. 高等学校化学学报, 2012, 33(6), 1527—1535)
[18]	Nayak R. R., Roy S., Dey J., Colloid Polym. Sci., 2006, 285(2), 219—224
[19]	van Nostrum C. F., Soft Matter, 2011, 7(7), 3246—3259
[20]	Zhu Z., Xu H., Liu H., González Y. I., Kaler E. W., Liu S., J. Phys. Chem. B, 2006, 110(33), 16309—16317
[21]	Abe M., Koike T., Nishiyama H., Sharma S. C., Tsubone K., Tsuchiya K., Sakai K., Sakai H., Shchipunov Y. A., Schmidt J., J. Colloid Interface Sci., 2009, 330(1), 250—253
[22]	Roy S., Dey J., Langmuir, 2003, 19(23), 9625—9629
[23]	Lee J. H., Danino D., Raghavan S. R., Langmuir, 2009, 25(3), 1566—1571
[24]	Gomes J. F. P. d. S., Sonnen A. F. P., Kronenberger A., Fritz J., Coelho M. Á. N., Fournier D., Fournier-Nöel C., Mauzac M., Winterhalter M., Langmuir, 2006, 22(18), 7755—7759
[25]	Yan X., An X., Nanoscale, 2013, 5(14), 6280—6283
[26]	Storrs R. W., Tropper F. D., Li H. Y., Song C. K., Kuniyoshi J. K., Sipkins D. A., Li K. C., Bednarski M. D., J. Am. Chem. Soc., 1995, 117(28), 7301—7306
[27]	Mueller A., O'Brien D. F., Chem. Rev., 2002, 102(3), 727—727
[28]	Puri A., Blumenthal R., Acc. Chem. Res., 2011, 44(10), 1071—1079
[29]	Chiaramoni N. S., Biophys. J., 2013, 104(2), 243a
[30]	Yavlovich A., Singh A., Tarasov S., Capala J., Blumenthal R., Puri A., J. Therm. Anal. Calorim., 2009, 98(1), 97—104
[31]	Jung Y. K., Kim T. W., Jung C., Cho D. Y., Park H. G., Small, 2008, 4(10), 1778—1784
[32]	Fan Y., Ma L., Fang Y., Li Y. J., Li Y. D., Fine Chem., 2013, 30(7), 729—733
	(樊晔, 马琳, 方云, 李宇婕, 李延頔. 精细化工, 2013, 30(7), 729—733)
[33]	Megias-Alguacil D., Tervoort E., Cattin C., Gauckler L. J., J. Colloid Interface Sci., 2011, 353(2), 512—518
[34]	Hanczyc M. M., Fujikawa S. M., Szostak J. W., Science, 2003, 302(5645), 618—622
[35]	Guo C., Liu S., Jiang C., Li W., Dai Z., Fritz H., Wu X., Langmuir, 2009, 25(22), 13114—13119
[36]	Qin G., Li Z., Xia R., Li F., O'Neill B. E., Goodwin J. T., Khant H. A., Chiu W., Li K. C., Nanotechnology, 2011, 22(15), 155605—155619
[37]	Becucci L., Papini M., Verardi R., Veglia G., Guidelli R., Soft Matter, 2012, 8(14), 3881—3888
[38]	Zhou Y. F., Yan D. Y., Angew. Chem. Int. Ed., 2004, 43(37), 4896—4899
[39]	Zhou Y.F., Yan D. Y.,Chem. Commun., 2009, (10), 1172—1188
[40]	Li X. J., Pivkin I. V., Liang H. J., Karniadakis G. E., Macromolecules, 2009, 42(8), 3195—3200
[41]	Li X. J., Soft Matter, 2009, 9(48), 11663—11670