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Preparing method and application of oral insulin transport system in layer-by-layer self-assembly structure

A layer-by-layer self-assembly and insulin technology, which is applied in the field of preparation of oral insulin delivery system, can solve the problems of insulin deficiency and poor insulin effect, and achieve the effects of low toxicity, control of blood sugar level, and small particle size

Active Publication Date: 2017-01-11
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some patients even produce too much insulin in the body, but the effect of insulin is poor, so the insulin in the patient's body is a kind of relative deficiency.

Method used

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  • Preparing method and application of oral insulin transport system in layer-by-layer self-assembly structure
  • Preparing method and application of oral insulin transport system in layer-by-layer self-assembly structure
  • Preparing method and application of oral insulin transport system in layer-by-layer self-assembly structure

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Example 1 Preparation and Characterization of Oral Insulin Delivery System (LBL) with Layer-by-Layer Self-Assembled Structure

[0088] 1. Preparation of LBL particles

[0089] The synthetic method of LBL particle is as attached figure 1 shown.

[0090] (1) Dissolve 5mg of insulin in 1ml of dilute hydrochloric acid solution with pH 1.1 at 15°C, then add solid NaCl to make the final concentration of NaCl reach 0.6M, stir the solution for 0.5h, and centrifuge at 5000rpm for 3min to obtain a precipitate;

[0091] (2) Add the polygalacturonic acid (PGLA) solution of pH = 4, NaCl 0.6M to the precipitate in step (1), stir for 0.5 h, sonicate for 1 min, continue stirring for 0.5 h, and centrifuge at 5000 rpm for 3 min to obtain the precipitate;

[0092] The preparation method of the polygalacturonic acid (PGLA) solution of described pH=4, NaCl 0.6M is: polygalacturonic acid (PGLA) is dissolved in 1ml dilute hydrochloric acid solution, then adds NaCl solid, makes NaCl final co...

Embodiment 2

[0121] Example 2 Cell experiment

[0122] 1. Experimental method

[0123] (1) Cell culture

[0124] 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, 5.0% CO 2 .

[0125] (2) Transport system toxicity

[0126] Different doses of the transport system were added to the Caco-2 cells being cultured, and after 24 hours, the cell viability was measured by the MTT method. That is, inoculate Caco-2 cells on a 96-well culture plate, 5 000 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, add 5mg / ml MTT solution 20ul / well. Cont...

Embodiment 3

[0130] Example 3 In Vivo Experiment—Studies on Cytotoxicity of the System

[0131] 1. Construction of type I diabetic mouse model

[0132] (1) Method

[0133] Purchase 6-week-old SD rats and adapt to the environment for one week. Take 3 as the control group, and the rest as the experimental group. Rats were fasted overnight, blood glucose was measured, and STZ solution was prepared with a concentration of 15 mg / ml by using citrate buffer solution with pH=4.4 in ice bath and protected from light before injection. Rats in the experimental group were injected intraperitoneally at a dose of 50 mg / kg.

[0134] From the day of modeling, the body weight, food intake and water intake of the rats were measured daily. On the 3rd and 7th day, the blood glucose was monitored with the Roche active blood glucose meter. For rats with low blood sugar, supplementary injection of STZ (30mg / kg).

[0135] One week later, those whose fasting (after 3 h of starvation) blood glucose concentrat...

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Abstract

The invention discloses a preparing method for oral insulin transport system in layer-by-layer self-assembly structure comprising the steps of separating out insulin nano ball with electropositivity by salting out method under the condition of low pH, and successively packing the insulin nano ball using polygalacturonic acid (PGLA) with electronegativity and chitosan (CS) with electropositivity by layer-by-layer self-assembly method according to the electrical property and electrostatic force of materials. The oral insulin transport system in the invention is characterized by nature, low toxicity, natural metabolism and degradation and excellent biocompatibility, and has small partical size, strong small-intestine absorption rate, high drug loading ratio and excellent stability. The system can quickly and potently exert the hypoglycemic effect of insulin and also obviously and quickly control the blood glucose level in a long-term with the potential of diabetes treatment.

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 an oral insulin delivery system with a layer-by-layer self-assembly structure. Background technique [0002] The number of people suffering from diabetes in the world is about 280 million, of which 5-10% are type I diabetes, and about 80-90% are type II diabetes. Type I diabetes is also known as juvenile diabetes, and 50% of the patients suffer from onset at the age of 0-4. Type I diabetes is caused by the death of pancreatic islet B cells due to immune system disorder. When the patient first develops, 70%-90% of pancreatic islet B cells have been lost. After 1 to 2 years of onset, all pancreatic islet B cells die, and the patient's insulin secretion is zero. Type II diabetes usually occurs between the ages of 35 and 40. Some patients even produce too much insulin in the body, but the effect of insulin is poo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/28A61K47/36A61P3/10A61K9/50A61K31/715
Inventor 关燕清张丽
Owner SOUTH CHINA NORMAL UNIVERSITY
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