Preparation method and application of a hydroxyapatite nano oral system grafted with insulin and gallic acid

A technology of hydroxyapatite and gallic acid, which is applied in the directions of medical preparations without active ingredients, medical preparations containing active ingredients, and pharmaceutical formulas

Active Publication Date: 2019-07-12
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although oral insulin is the best choice for diabetic patients, how to find a method that is simple, stable, low toxicity, natural ingredients, good biocompatibility, drug loading rate, high release rate, good absorption effect and Oral drugs that play a role in diabetes complications are still a problem

Method used

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  • Preparation method and application of a hydroxyapatite nano oral system grafted with insulin and gallic acid
  • Preparation method and application of a hydroxyapatite nano oral system grafted with insulin and gallic acid
  • Preparation method and application of a hydroxyapatite nano oral system grafted with insulin and gallic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Example 1 Preparation of a nano oral delivery system carrying insulin and gallic acid

[0087] 1. Preparation of hydroxyapatite (HAP) nanoparticles

[0088] (1) Take Ca(NO 3 ) 2 · 4 h 2 O and (NH 4 ) 2 HPO 4 Dissolve in ultrapure water respectively, adjust the pH to 11 with ammonia water, and then add to the mixing system.

[0089] Among them, the Ca(NO 3 ) 2 · 4 h 2 O: (NH 4 ) 2 HPO 4 =1.67mol: 1mol.

[0090] The mixed system is a system in which the volume ratio of Triton X-100:cyclohexane:n-butanol=16ml:56ml:6ml.

[0091] (2) Mix the two solutions of step (1) for about 20 minutes to form a transparent microemulsion, add 30ml of absolute ethanol to the mixed microemulsion to break the emulsion, and centrifuge at 12000g for 15min to remove cyclohexane and triton X-100 (surfactant), washed with absolute ethanol for 2 to 3 times to obtain granules.

[0092] (3) Put the particles obtained in step (2) in an oven and dry them at 80° C. for 4 hours to obtain...

Embodiment 2

[0100] Example 2 Characterization of the nano oral delivery system carrying insulin and gallic acid

[0101] 1. Particle size detection

[0102] (1) The prepared HAP nanoparticles were dissolved in acetone, 80 μl was added to the sample pool, and detected by a Malvern particle size detector.

[0103] (2) figure 1 It is the detection result of the particle size of HAP nanoparticles prepared by inverse microemulsion method, PEG-wrapped HAP, GA connected to PEG and finally INS connected to PEG by Malvern particle size detector.

[0104] In the particle size detection chart, figure a shows that the average particle size of HAP nanoparticles prepared by inverse microemulsion method is about 100-200nm; figure b shows that the average particle size of HAP after PEG wrapping is about 300nm; figure c shows that GA is connected to PEG In the future, the particle size will be less than 200nm; Figure d shows that after INS is connected to PEG, the particle size will be around 500nm.

...

Embodiment 3

[0120] Example 3 Cell experiment

[0121] 1. MTT cytotoxicity test

[0122] (1) Collect Caco-2 cells in the logarithmic phase, adjust the concentration of the cell suspension to 5,000-10,000 cells / well, incubate until the cell monolayer covers the bottom of the well, and add drugs with a concentration gradient. Incubate for 18-24 hours and observe under an inverted microscope. Add MTT, terminate the culture, add dimethyl sulfoxide, and use untreated blank cells as a control. The cell viability was obtained by a microplate reader.

[0123] (2) Image 6 It shows that in the MTT method to explore the toxicity of HAP-PEG-GA-INS nanoparticles to Caco-2 cells, the experiment was set within the concentration gradient range of nanoparticles from 0 to 100 μg / ml, and the particles with a concentration of 0 μg / ml were used as the control group , the cell survival rate was set as 100%, and the particle concentration of 12.5, 25, 50 and 100 μg / ml was added as the experimental group, an...

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Abstract

The invention discloses a preparing method of hydroxyapatite nanoparticles oral system for grafting insulin and gallic acid, comprising the following steps: firstly to synthetize hydroxyapatite (HAP) nanoparticles with microporous morphology, and then tightly wrap HAP surface with PEG, after that, connect gallic acid (GA) to the hydroxyl group of PEG, and finally connect insulin to the PEG surface, to get the nanometer oral transportation system carrying insulin and gallic acid, namely, HAP-PEG- GA-INS nanoparticle. The nano oral transportation system has good stability, natural ingredients, good biocompatibility, high drug loading ratio and release rate, good absorbing effect, the endurance capacity is strong in the gastrointestinal fluid, and absorption is significant in the small intestine epithelial cell line, low cytotoxicity, with significant hypoglycemic effect through oral administration, and has good application potential on diabetes treatment and drug research.

Description

technical field [0001] The invention belongs to the field of biomedical materials. More specifically, it relates to the preparation and application of a hydroxyapatite nano oral system grafted with insulin and gallic acid. Background technique [0002] Diabetes mellitus (DM): Diabetes mellitus is a metabolic disease characterized by hyperglycemia. Hyperglycemia is caused by defective insulin secretion or impaired biological action, or both. The long-term high blood sugar in diabetes leads to chronic damage and dysfunction of various tissues, especially the eyes, kidneys, heart, blood vessels, and nerves. Diabetes is one of the most important chronic non-communicable diseases threatening human health all over the world. As of November 2014, the number of patients worldwide has reached 382 million. Diabetes mellitus is a polygenic hereditary disease, and insulin deficiency and insulin resistance are its main pathological features. Commonly used methods for treating diabet...

Claims

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Application Information

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IPC IPC(8): A61K38/28A61K47/60A61K47/52A61K47/69A61P3/10A61K31/192
CPCA61K9/0053A61K31/192A61K38/28A61K2300/00
Inventor 关燕清张豫骁张丽
Owner SOUTH CHINA NORMAL UNIVERSITY
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