去氧胆酸修饰的聚合物胶束的构建及其跨膜转运的研究

doi:10.5246/jcps.2021.01.002

中国药学(英文版) ›› 2021, Vol. 30 ›› Issue (1): 17-26.DOI: 10.5246/jcps.2021.01.002

去氧胆酸修饰的聚合物胶束的构建及其跨膜转运的研究

刘琦, 王乐淇, 胡新平, 周楚杭, 唐颖蔚, 马翊宁, 王骁潇, 刘艳^*()

北京大学药学院分子药剂学与新释药系统北京市重点实验室, 北京 100191

收稿日期:2020-07-07 修回日期:2020-08-13 接受日期:2020-08-26 出版日期:2021-01-29 发布日期:2021-01-29
通讯作者: 刘艳
作者简介:
+ Tel.: +86-10-82801508, E-mail: yanliu@bjmu.edu.cn
基金资助:
The National Natural Science Foundation of China (Grant No. 81673366).

Fabrication of deoxycholic acid-modified polymeric micelles and their transmembrane transport

Qi Liu, Leqi Wang, Xinping Hu, Chuhang Zhou, Yingwei Tang, Yining Ma, Xiaoxiao Wang, Yan Liu^*()

Beijing Key Laboratory of Molecular Pharmaceutics and New Drug Delivery Systems, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China

Received:2020-07-07 Revised:2020-08-13 Accepted:2020-08-26 Online:2021-01-29 Published:2021-01-29
Contact: Yan Liu

摘要/Abstract

摘要：

口服给药是患者顺应性最好的给药方式, 而小肠上皮是口服药物吸收的主要屏障。为了克服小肠上皮屏障口服递送难溶性药物, 本研究设计合成了小肠胆酸转运体的底物脱氧胆酸偶联的聚(2-乙基-2-噁唑啉)-聚(D,L-乳酸)(DA-PEOz-PLA), 并基于小肠胆酸特殊的转运途径构建了由DA-PEOz-PLA和mPEG-PLA组成的聚合物胶束以包载模型药物香豆素6(C6)。用核磁共振氢谱和薄层色谱对DA-PEOz-PLA的结构进行了确证, 用凝胶渗透色谱测得其分子量为10 034 (PDI = 1.51)。载C6的聚合物胶束的粒径和PDI分别为40.11 nm和0.212, 载药量为0.085%。透射电镜观察胶束呈现较规则的球形。另外, 与未修饰胶束相比, 去氧胆酸修饰的聚合物胶束能进一步增强胶束的跨膜转运效率。跨膜转运机制的研究结果表明去氧胆酸的修饰丰富了胶束的跨膜转运途径。因此, 所制备的去氧胆酸修饰的聚合物胶束在难溶性药物的口服递送方面具有潜在的优势。

关键词: 去氧胆酸, 小肠胆酸转运体, 聚合物胶束, 口服纳米给药系统, 跨膜转运

Abstract:

Oral administration is the best way for the most patients due to the good compliance, and intestinal epithelium is the main barrier of oral drug absorption. In order to overcome the small intestine epithelial barrier to orally deliver water-insoluble drugs, deoxycholic acid (DA), a substrate of the intestinal bile acid transporters, conjugated poly(2-ethyl-2-oxazoline)-poly(D,L-lactide) (DA-PEOz-PLA) was designed and synthesized, and deoxycholic acid-modified polymeric micelles composed of DA-PEOz-PLA and mPEG-PLA were fabricated to encapsulate model drug coumarin 6 (C6) based on intestinal bile acid pathway. The structure of DA-PEOz-PLA was confirmed using 1H NMR and TLC, and the molecular weight measured by GPC was 10 034 g/mol with a PDI of 1.51. The C6-loaded polymeric micelles with drug loading content of 0.085% were characterized to have 40.11 nm in diameter and uniform spherical morphology observed by TEM. Furthermore, the deoxycholic acid-modified polymeric micelles were demonstrated to further enhance the transmembrane transport efficiency. The mechanic study evidenced that anchorage of deoxycholic acid onto the micelles surface enriched their transcellular transport pathway. Therefore, the designed deoxycholic acid-modified polymeric micelles might have a promising potential for oral delivery of water-insoluble drugs.

Key words: Deoxycholic acid, Intestinal bile acid transporter, Polymeric micelles, Oral nano drug delivery system, Transmembrane transport

Supporting:

刘琦, 王乐淇, 胡新平, 周楚杭, 唐颖蔚, 马翊宁, 王骁潇, 刘艳. 去氧胆酸修饰的聚合物胶束的构建及其跨膜转运的研究[J]. 中国药学(英文版), 2021, 30(1): 17-26.

Qi Liu, Leqi Wang, Xinping Hu, Chuhang Zhou, Yingwei Tang, Yining Ma, Xiaoxiao Wang, Yan Liu. Fabrication of deoxycholic acid-modified polymeric micelles and their transmembrane transport[J]. Journal of Chinese Pharmaceutical Sciences, 2021, 30(1): 17-26.

图/表 6

Figure 1. Synthesis routes of HOOC-PEOz-PLA (A), NH2-PEOz-PLA (B) and DA-PEOz-PLA (C).

Figure 2. 1H NMR spectra of HOOC-PEOz-OH (A), HOOC-PEOz-PLA (B) in CDCl3 and DA-PEOz-PLA (C) in DMSO-d6.

Figure 3. The GPC spectrum of HOOC-PEOz-OH (A) and HOOC-PEOz-PLA (B).

Figure 4. The thin layer chromatograms of HOOC-PEOz-PLA (PP), NH2-PEOz-PLA (PP-NH2) and ethylenediamine (eda) were developed to be yellow spots by iodine (A) and developed by ninhydrin (B). (C) Thin layer chromatograms of DA-PEOz-PLA (DA-PP), NH2-PEOz-PLA (PP-NH2) and deoxycholic acid (DA) were developed by phosphomolybdic acid.

Figure 5. (A) Schematic diagram of the fabrication for C6-loaded deoxycholic acid-conjugated micelles C6/DA-MIX-PP-PMs. (B) Size distribution and (C) TEM micrograph of C6/DA-MIX-PP-PMs.

Figure 6. (A) Apparent permeability coefficient Papp of C6/PP-PMs and C6/DA-MIX-PP-PMs across Caco-2 cell monolayer. Transmembrane transport of C6/PP-PMs (B) and C6/DA-MIX-PP-PMs (C) in the presence of different inhibitors (n = 3). nsP > 0.05, *P < 0.05, **P < 0.01.

参考文献 19

[1]	Gaucher, G.; Satturwar, P.; Jones, M.C.; Furtos, A.; Leroux, J.C. Polymeric micelles for oral drug delivery. Eur. J. Pharm. Biopharm. 2010, 76, 147–158.
[2]	Deng, Y.Q. Preparation and characterization of intestine PepT1-targeted calcium carbonate nanoparticles. J. Chin. Pharm. Sci. 2018, 27, 397–407.
[3]	Savjani, K.T.; Gajjar, A.K.; Savjani, J.K. ChemInform abstract: drug solubility: importance and enhancement techniques. ChemInform. 2013, doi: 10.1002/chin.201326246.
[4]	Kalepu, S.; Nekkanti, V. Insoluble drug delivery strategies: review of recent advances and business prospects. Acta Pharm. Sin. B. 2015, 5, 442–453
[5]	Reddy, M.S.; Gurram, A.K.; Deshpande, P.B.; Kar, S.S.; Nayak, U.; Udupa, N. Role of components in the formation of self-microemulsifying drug delivery systems. Indian J. Pharm. Sci. 2015, 77, 249.
[6]	Pridgen, E.M.; Alexis, F.; Farokhzad, O.C. Polymeric nanoparticle drug delivery technologies for oral delivery applications. Expert. Opin. Drug Deliv. 2015, 12, 1459–1473.
[7]	Atuma, C.; Strugala, V.; Allen, A.; Holm, L. The adherent gastrointestinal mucus gel layer: thickness and physical state in vivo. Am. J. Physiol. Gastrointest. Liver Physiol. 2001, 280, G922–G929.
[8]	Varma, M.V.S.; Ambler, C.M.; Ullah, M.; Rotter, C.J.; Sun, H.; Litchfield, J.; Fenner, K.S.; El-Kattan, A. Targeting intestinal transporters for optimizing oral drug absorption. Curr. Drug Metab. 2010, 11, 730–742.
[9]	Jin, Y.; Liu, Q.; Zhou, C.H.; Hu, X.P.; Wang, L.Q.; Han, S.D.; Zhou, Y.H.; Liu, Y. Intestinal oligopeptide transporter PepT1-targeted polymeric micelles for further enhancing the oral absorption of water-insoluble agents. Nanoscale. 2019, 11, 21433–21448.
[10]	He, C.Y. Preparation and characterization of intestinal transporter-targeted polymeric micelles. J. Chin. Pharm. Sci. 2018, 27, 490–497.
[11]	Alrefai, W.A.; Gill, R.K. Bile acid transporters: structure, function, regulation and pathophysiological implications. Pharm. Res. 2007, 24, 1803–1823.
[12]	Balakrishnan, A.; Polli, J.E. Apical sodium dependent bile acid transporter (ASBT, SLC10A2): a potential prodrug target. Mol. Pharmaceutics. 2006, 3, 223–230.
[13]	Chiang, J.Y. Bile Acid Metabolism and Signaling. Compr Physiol. 2013, 3, 1191–1212.
[14]	Park, J.W.; Jeon, O.C.; Kim, S.K.; Al-Hilal, T.A.; Jin, S.J.; Moon, H.T.; Yang, V.C.; Kim, S.Y.; Byun, Y. High antiangiogenic and low anticoagulant efficacy of orally active low molecular weight heparin derivatives. J. Control. Release. 2010, 148, 317–326.
[15]	Fan, W.W.; Xia, D.N.; Zhu, Q.L.; Li, X.Y.; He, S.F.; Zhu, C.L.; Guo, S.Y.; Hovgaard, L.; Yang, M.S.; Gan, Y. Functional nanoparticles exploit the bile acid pathway to overcome multiple barriers of the intestinal epithelium for oral insulin delivery. Biomaterials. 2018, 151, 13–23.
[16]	Park, J.; Choi, J.U.; Kim, K.; Byun, Y. Bile acid transporter mediated endocytosis of oral bile acid conjugated nanocomplex. Biomaterials. 2017, 147, 145–154.
[17]	Fan, W.W.; Xia, D.N.; Zhu, Q.L.; Hu, L.; Gan, Y. Intracellular transport of nanocarriers across the intestinal epithelium. Drug Discov. Today. 2016, 21, 856–863.
[18]	He, B.; Lin, P.; Jia, Z.R.; Du, W.W.; Qu, W.; Yuan, L.; Dai, W.B.; Zhang, H.; Wang, X.Q.; Wang, J.C.; Zhang, X.; Zhang, Q. The transport mechanisms of polymer nanoparticles in Caco-2 epithelial cells. Biomaterials. 2013, 34, 6082–6098.
[19]	Annaba, F.; Sarwar, Z.; Kumar, P.; Saksena, S.; Turner, J.R.; Dudeja, P.K.; Gill, R.K.; Alrefai, W.A. Modulation of ileal bile acid transporter (ASBT) activity by depletion of plasma membrane cholesterol: association with lipid rafts. Am. J. Physiol. Gastrointest. Liver Physiol. 2008, 294, G489–G497.

去氧胆酸修饰的聚合物胶束的构建及其跨膜转运的研究

Fabrication of deoxycholic acid-modified polymeric micelles and their transmembrane transport

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