Fluoride-Functionalized Choline Phosphate Liposomes for Oral Insulin Administration

doi:10.19894/j.issn.1000-0518.220384

Chinese Journal of Applied Chemistry ›› 2023, Vol. 40 ›› Issue (5): 730-742.DOI: 10.19894/j.issn.1000-0518.220384

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Fluoride-Functionalized Choline Phosphate Liposomes for Oral Insulin Administration

Yuan-Hua ZHANG¹^,², Sheng-Ran LI¹, Xi-Fei YU¹^,²()

^1.State Key Laboratory of Polymer Physics and Chemistry，Changchun Institute of Applied Chemistry，Chinese Academy of Science，Changchun 130022，China
^2.University of Science and Technology of China，Hefei 230026，China

Received:2022-11-23 Accepted:2023-03-27 Published:2023-05-01 Online:2023-05-26
Contact: Xi-Fei YU
About author:xfyu@ciac.ac.cn
Supported by:
the National Natural Science Foundation of China(21674109)

Abstract

Abstract:

Subcutaneous insulin （INS） injection as the main therapy method for diabetes shows adverse effects such as inconvenient use and hypoglycemia. Fortunately， the oral route of administration of insulin possesses more application potential. As one of the most mature drug delivery systems， liposomes exist some disadvantages in the oral administration of proteins or peptides， such as poor stability， low drug load， and limited mucus penetration. Herein， according to the phosphocholine （PC） component of the cell membrane， choline phospholipid with trifluoroethyl modified zwitterionic head （DPFCP） is designed and synthesized for constructing liposomes delivery system loaded with insulin （INS@DPFCP） for oral administration. By comparison with phosphate choline liposomes （INS@DPPC） and unfunctionalized choline phosphate liposomes （INS@DPCP）， the physicochemical properties， drug release stability， mucus permeability， biocompatibility， endocytosis， and transport ability of INS@DPFCP are studied. The test results indicate that the particle size of INS@DPFCP is 185 nm， and drug entrapment efficiency is （47.1±3.9）%. Compared with INS@DPPC （27.2%） and INS@DPCP （24.5%）， the amount of mucus aggregation for INS@DPFCP is 17.15%. Hemolysis and cytotoxicity experiments show that INS@DPFCP gets good biocompatibility. When the mass concentration is as high as 1.2 mg/mL， the hemolysis rate is 6.88%， and the cell viability is above 90%. In addition， the endocytosis assay results demonstrate that INS@DPFCP is proved to be higher cellular uptake efficiency of Caco-2 cells， and the drug delivery accumulation reaches 47.5% in 4 h. INS@DPFCP is expected to play an important role in diabetes treatment， and shows potential application in the field of proteins or peptides oral administration.

Key words: Choline phosphate liposomes, Fluoride-functionalized, Insulin, Oral administration

CLC Number:

O623

Yuan-Hua ZHANG, Sheng-Ran LI, Xi-Fei YU. Fluoride-Functionalized Choline Phosphate Liposomes for Oral Insulin Administration[J]. Chinese Journal of Applied Chemistry, 2023, 40(5): 730-742.

Figures/Tables 8

Fig.1 Schematic illustration of the molecular structures of DPPC， DPCP and DPFCP lipids and their insulin-loaded liposomes

Fig 2 The synthesis routes of DPFCP

Fig.3 The 1H， 31P and 19F NMR （A）， and ESI-MS （B） spectra of DPFCP

Fig.4 （A） The size distribution （pH=7.4） of INS@DPPC， INS@DPCP， DPFCP and INS@DPFCP liposomes；（B） Size change and （C） Zeta potential change in different pH （1.0， 2.0， 4.0， 6.0， 7.4 and 9.0） solution for 2 h of DPPC， DPCP， DPFCP and INS@DPFCP liposomes；（D） TEM images of INS@DPFCP liposomes；（E） Tm of DPPC， DPCP and DPFCP

Table 1 The size， PDI， Zeta potential， DLE and DLC of INS@DPPC， INS@DPCP， DPFCP and INS@DPFCP liposomes

Sample	Size/nm	PDI	Zeta potential/mV	DLE/%	DLC/%
INS@DPPC	157	0.219	-22.1±1.8	45.2±2.3	3.39±0.41
INS@DPCP	169	0.223	-24.6±1.2	42.7±1.2	3.2±0.68
DPFCP	163	0.236	-10.3±2.5	/	/
INS@DPFCP	185	0.247	-18.9±2.7	47.1±3.9	3.53±0.35

Fig.5 （A） The cumulative release and （B） the enzymatic degradation in SGF（0~2 h） and SIF（2~6 h） of INS@DPPC， INS@DPCP and INS@DPFCP liposomes in PBS buffer solution at 37 ℃；（C） CD curves of standard insulin and released insulin of INS@DPFCP；（D） Changes in size and encapsulation efficiency of INS@DPFCP liposomes at 4 ℃ for 20 d；（E） Aggregation and （F） Papp of DPCP-CTAC， DPPC， DPCP and DPFCP in mucin solution for 2 h

Fig.6 （A， B） Hemolysis of different concentrations of DPPC， DPCP and DPFCP liposomes after 2 h incubation with mouse RBC； The cell viability to Caco-2 cells （C） and NIH-3T3 cells （D） fed by DPPC， DPCP and DPFCP liposomes for 24 h

Fig.7 Flow cytometric analysis in treated Caco-2 cells with Free INS， INS@DPPC， INS@DPCP and INS@DPFCP liposomes for （A） 2 h and （B） 4 h， and （C） CLSM images for 4 h， scale bar is 20 μm；（D） The cumulative transport volume and （E） Papp of the monolayer cell membrane model of Caco-2 cells for 4 h

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Fluoride-Functionalized Choline Phosphate Liposomes for Oral Insulin Administration

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