Method for preparing nano oral protein drug delivery system and application thereof

A protein and drug technology, applied in the field of preparation of nanometer oral protein drug delivery system, can solve problems such as enhancing therapeutic effect

Active Publication Date: 2016-11-23
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the current oral protein delivery system only considers the improvement of delivery efficiency, and does not achieve simultaneous enhancement of therapeutic effect

Method used

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  • Method for preparing nano oral protein drug delivery system and application thereof
  • Method for preparing nano oral protein drug delivery system and application thereof
  • Method for preparing nano oral protein drug delivery system and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] Example 1 Preparation of nanometer oral protein drug delivery system

[0098] Synthesize nanoparticles (nano oral protein drug delivery system) by W / O / W double emulsification method, as attached figure 1 shown.

[0099] Specifically, the preparation method of the nano oral protein drug delivery system is as follows:

[0100] 1. Preparation of insulin-loaded PGLA nanoparticles (PGLA NP)

[0101] (1) Prepare the inner aqueous phase:

[0102] Add PGLA to 5% Pluronic 188, adjust to pH=4, incubate at 50°C for 30min, shake, dissolve for 24h, centrifuge to remove insoluble precipitate, take 200μl of the obtained solution, add 1mg bovine insulin to dissolve; the PGLA is mixed with 5% (w / v) Poloxamer 188 has a mass volume ratio of 10 mg / ml.

[0103] (2) Prepare PGLA NP with ethyl acetate as the oil phase and 20% (w / v) Pluronic 188 as the external aqueous phase solution:

[0104] Add the internal water phase of step (1) into 1ml of ethyl acetate, and ultrasonicate for 60s to...

Embodiment 2

[0114] The characterization of embodiment 2 nano oral protein drug delivery system

[0115] 1. Infrared spectroscopy detection (FTIR)

[0116] (1) The raw materials PGLA, chitosan, insulin, genipin and the three systems synthesized by the system were detected by infrared spectroscopy. Freeze-dry PGLA NP, CS-PGLA NP, and Gen-CS-PGLA NP, then put them into a mortar, add a certain amount of KBr, and grind the mixture evenly until the particle size is less than 2 μm, so as not to be affected by scattered light, then put it into a dry Machine drying. Press the mixture into a transparent sheet with a pressure of about 10 MPa on a hydraulic press, and use a German Bruker VERTEX 33 Fourier transform infrared spectrometer to measure it.

[0117] (2) Infrared spectroscopy analysis can explain to a certain extent the composite components of the synthesized nanosystems ( figure 2 A). The infrared spectra of PGLA NP, CS-PGLA NP and Gen-CS-PGLA NP show that the three are at 1100cm -1 ...

Embodiment 3

[0149] Example 3 Cell Experiment of Nano Oral Protein Drug Delivery System

[0150] 1. Cell culture

[0151] Caco-2 cell line was provided by Guangdong Pharmaceutical University. After the cells were proliferated and cultured to 80% in the culture flask, they were seeded on a 96-well plate at a density of 5000 / well, and cultured for 1 or 2 days for subsequent experiments. The cell culture conditions are: high-glucose DMEM medium containing 20% ​​newborn calf serum, non-essential amino acids, 37°C constant temperature, 5.0% CO 2 .

[0152] 2. Cytotoxicity test of the delivery system

[0153] (1) Add different doses of the delivery system to the Caco-2 cells being cultured, and after 24 hours, use the MTT method to measure the cell survival rate. That is to inoculate Caco-2 cells on a 96-well culture plate, 5000 cells / well, after 24 hours of culture, aspirate the medium, add serum-free medium, and different concentrations of insulin delivery system, after 24 hours of culture...

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Abstract

The invention discloses a method for preparing a polygalacturonic acid-based nano oral protein drug delivery system and application thereof. The system is prepared by adopting a W / O / W double emulsion method. The method comprises the following steps: dissolving polygalacturonic acid in a 4-6w / v% poloxamer 188 solution, and adding bovine insulin for dissolving, thereby obtaining an inner water phase; taking ethyl acetate as an oil phase, respectively taking 18-22w / v% poloxamer 188 solution, 18-22w / v% poloxamer 188 solution dissolved with chitosan and 18-22w / v% poloxamer 188 solution dissolved with chitosan and genipin as outer water phase solutions, and preparing to obtain different nano oral protein drug delivery systems. The nano oral protein drug delivery system disclosed by the invention is non-toxic, easy to degrade, high in delivery rate and high in absorption rate, and has high application prospects in the fields such as delivery research and application of oral protein drugs such as insulin, diabetes treatment, and vaccine administration.

Description

technical field [0001] The invention belongs to the technical field of biomedical materials. More specifically, it relates to a preparation method and application of a nano oral protein drug delivery system. Background technique [0002] With the development of biomedical research, more and more protein drugs have been developed. Due to the instability of protein drugs, injections are usually used to enter the human body to exert therapeutic effects. However, the injection of the drug poses many problems. For example, the injection of protein vaccines is often administered to young children, and the pain and resistance caused will lead to inaccurate drug injection doses and increase the probability of infection. Insulin and other protein hormone drugs require frequent and long-term injections, which increase the suffering of patients, cause local necrosis or hyperplasia of the epidermis, and increase the probability of infection. At the same time, the distribution concent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/48A61K38/28A61K31/715A61K9/10A61P3/10
CPCA61K9/1075A61K31/715A61K38/28A61K2300/00
Inventor 关燕清张丽
Owner SOUTH CHINA NORMAL UNIVERSITY
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