Overcoming oral insulin delivery barriers: application of cell penetrating peptide and silica-based nanoporous composites

doi:10.1007/s11705-013-1306-9

Front Chem Sci Eng

2013, Vol. 7

Issue (1) : 9-19 https://doi.org/10.1007/s11705-013-1306-9

REVIEW ARTICLE

Overcoming oral insulin delivery barriers: application of cell penetrating peptide and silica-based nanoporous composites

Huining HE^1,4,5, Junxiao YE¹, Jianyong SHENG², Jianxin WANG², Yongzhuo HUANG^2,3, Guanyi CHEN⁴, Jingkang WANG¹(

), Victor C YANG^5,6(

)

1. School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China; 2. Department of Pharmaceutics, School of Pharmacy, Fudan University; Key Laboratory of Smart Drug Delivery, Ministry of Education & PLA, Shanghai 201203, China; 3. Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China; 4. School of Environmental Science and Engineering, State Key Laboratory of Engines, Tianjin University, Tianjin 300072, China; 5. Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China; 6. Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, Michigan 48109-1065, USA

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Abstract

Oral insulin delivery has received the most attention in insulin formulations due to its high patient compliance and, more importantly, to its potential to mimic the physiologic insulin secretion seen in non-diabetic individuals. However, oral insulin delivery has two major limitations: the enzymatic barrier that leads to rapid insulin degradation, and the mucosal barrier that limits insulin’s bioavailability. Several approaches have been actively pursued to circumvent the enzyme barrier, with some of them receiving promising results. Yet, thus far there has been no major success in overcoming the mucosal barrier, which is the main cause in undercutting insulin’s oral bioavailability. In this review of our group’s research, an innovative silica-based, mucoadhesive oral insulin formulation with encapsulated-insulin/cell penetrating peptide (CPP) to overcome both enzyme and mucosal barriers is discussed, and the preliminary and convincing results to confirm the plausibility of this oral insulin delivery system are reviewed. In vitro studies demonstrated that the CPP-insulin conjugates could facilitate cellular uptake of insulin while keeping insulin’s biologic functions intact. It was also confirmed that low molecular weight protamine (LMWP) behaves like a CPP peptide, with a cell translocation potency equivalent to that of the widely studied TAT. The mucoadhesive properties of the produced silica-chitosan composites could be controlled by varying both the pH and composition; the composite consisting of chitosan (25 wt-%) and silica (75 wt-%) exhibited the greatest mucoadhesion at gastric pH. Furthermore, drug release from the composite network could also be regulated by altering the chitosan content. Overall, the universal applicability of those technologies could lead to development of a generic platform for oral delivery of many other bioactive compounds, especially for peptide or protein drugs which inevitably encounter the poor bioavailability issues.

Keywords insulin cell penetrating peptide mucoadhesive composites oral delivery

Corresponding Author(s): WANG Jingkang,Email:wangjkch@tju.edu.cn; YANG Victor C,Email:vcyang@umich.edu

Issue Date: 05 March 2013

Cite this article:

Huining HE,Junxiao YE,Jianyong SHENG, et al. Overcoming oral insulin delivery barriers: application of cell penetrating peptide and silica-based nanoporous composites[J]. Front Chem Sci Eng, 2013, 7(1): 9-19.

URL:

https://academic.hep.com.cn/fcse/EN/10.1007/s11705-013-1306-9
https://academic.hep.com.cn/fcse/EN/Y2013/V7/I1/9

Fig.1 Schematic diagrams of (a) silica/alginate-based mucoadhesive nanocomposite loaded with insulin-CPP conjugates, (b) mechanism of action of the proposed oral insulin delivery system in allowing insulin to target intestinal mucosa and cross through the epithelial layer (Reprinting with permission)

Fig.2 Synthesis and characterization of the insulin-TAT conjugates. (a) Insulin was linked to TAT using SMCC as a crosslinker; (b) SDS-PAGE analysis of protein bands either stained with Coomassie brilliant blue (left panel) or visualized with UV light (right 2 panels) prior to HPLC (left two panels) and after HPLC (right panel) purification, where lane 1 is free insulin and lane 2 is insulin-TAT conjugates; (c) elution profile of insulin, insulin-TAT conjugates, and TAT using a heparin affinity column (Reprinting with permission)

Fig.3 Fluorescence microphotographs after exposure of Caco-2 cells to (a) FITC-labeled free insulin or (b) insulin-TAT conjugates. (c) In vitro intestinal absorption of the insulin-TAT conjugates. Both apical-to-basal and basal-to-apical absorption by Caco-2 cell monolayers of FITC-labeled insulin and insulin-TAT conjugates were examined. Samples were collected every 2 h and analyzed by fluorescence spectroscopy (Reprinting with permission)

Fig.4 Integrity of insulin after translocation of insulin-TAT conjugates. The results from ELISA were plotted against the results from fluorescent assay. Intact (integrity) and enzyme degraded (degradation) insulin-TAT were used as negative and positive controls, respectively. Two samples (1 and 2) were collected from the receiver chamber of the transwell culture plates (Reprinting with permission)

Tab.1 Amino acid sequences for TAT and LMWP

Fig.5 Cellular translocation of LMWP-FITC conjugates into cultured HeLa cells after 30 min of in vitro incubation (left: differential interference contrast image; right: fluorescence microscopy) (Reprinting with permission)

Fig.6 (a) Cellular localization of rhodamine-labeled phalloidin-LMWP conjugates into MG63 osteoblast cells; (b) flow cytometric analysis of AlexFluor-488 labeled phalloidin, LMWP-phalloidin, and TAT-phalloidin in MG63 osteoblast cells after incubation at 37°C for 30 min in a medium containing 10% serum (Reprinting with permission)

Fig.7 (a) Tumor regression after intra-tumoral injection of LMWP-gelonin or TAT-gelonin conjugates. Tumor volumes were measured 30 days after cessation of drug treatment. Right: top to bottom represent tumors excised from mice treated with PBS solution (average tumor mass: 3.16±0.65 g); 100 μg gelonin (2.62±0.53 g); 110 μg LMWP-gelonin (0.33±0.12 g); 10 μg LMWP physically mixed with 100 μg gelonin (2.74±0.68 g); and 110 μg TAT-gelonin (0.21±0.04 g). (b) Tumor penetration of rhodamine-labeled free gelonin (top) or LWMP-gelonin conjugates (bottom) into the subcutaneously implanted tumor of a mouse. Ten hours after injection of the labeled drug or drug-conjugates, tumors were excised, processed into cryosections and examined by confocal microscopy. Left and right panels represent fluorescence microphotographs and DIC images, respectively (Reprinting with permission)

Fig.8 Effects of pH and silica content on the mucoadhesive properties of the nanocomposites. Mucoadhesion studies were performed via the rotating cylinder method in either HCl buffer (pH 2.0) or 100 mmol?L PBS buffer (pH 7.4) at 37°C. Data was presented as means±standard deviation of three experiments (Reprinting with permission)

Fig.9 Effect of chitosan content on the release of amoxicillin from the silica-chitosan composites (Reprinting with permission)

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