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A kind of nano-drug carrier and its combination medicine for the treatment of diabetes

A nanoparticle, gene recombination technology, applied in the intersection of bionics and biomedicine, in the field of nanobiology

Active Publication Date: 2020-02-14
CHINA SCI XINYUN BIOTECH BEIJING CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, because it cannot deliver insulin "on time" with blood sugar changes like normal islet cells, and regulate blood sugar in time, it will also cause a series of complications

Method used

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  • A kind of nano-drug carrier and its combination medicine for the treatment of diabetes
  • A kind of nano-drug carrier and its combination medicine for the treatment of diabetes
  • A kind of nano-drug carrier and its combination medicine for the treatment of diabetes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 After ferritin is mixed with insulin, stable insulin-ferritin nanoparticles can be formed

[0029] In order to study whether ferritin and insulin can interact with each other to form stable nanoparticles, recombinant human H-ferritin (prepared by the inventor, see invention patent ZL201110122433.0) and human recombinant insulin (91077c, Sigma) were selected for Research. In vitro, firstly, the present invention proves that ferritin and insulin are mixed in a certain ratio to form stable nanoparticles; and the insulin-ferritin nanoparticles are characterized.

[0030] The experimental method is as follows:

[0031] 1.1 Preparation of insulin-ferritin nanoparticles.

[0032] Ferritin and insulin were mixed in 0.2M concentration of PBS (137mM NaCl, 2.7mM KCl, 10mM NaCl at a molar ratio of 1:1 2 HPO4,2mM KH 2 PO4, pH 7.4) buffer and incubated at 4°C for 45 minutes. Then, by molecular exclusion, the Superdex 200 10 / 300GL molecular sieve purification method was...

Embodiment 2

[0036] Example 2 Insulin-ferritin nanoparticles can realize the slow release of insulin

[0037] After insulin-ferritin self-organizes into stable particles, the first question that needs to be verified is whether the assembled insulin can be effectively released slowly and exert its activity. The invention proves for the first time that insulin and ferritin can be self-assembled into stable nanoparticles based on the hydrophilic-hydrophobic interaction of the surface residues of the two proteins, and can release insulin slowly. The released insulin remains highly biologically active.

[0038] The specific experimental method is as follows:

[0039] 2.1 Slow release curve of insulin-ferritin in PBS

[0040] To evaluate the ability of insulin-ferritin nanoparticles to release insulin slowly, the inventors performed an insulin release experiment. First, using known concentration of insulin, using HPLC technology, draw the A280 value of the peak at a specific position on the HPL...

Embodiment 3

[0046] Example 3 Insulin-ferritin sustained release system can effectively reduce blood sugar level

[0047] In order to directly evaluate the reduction of blood glucose level by insulin-ferritin nanoparticles, the inventors conducted a blood glucose reduction experiment at the animal level. The experimental animals were male Sprague-Dawley rats, weighing 200-250 g, purchased from Beijing Huafukang Biotechnology Co., Ltd. (China). The experimental animals were divided into 3 groups (6 animals in each group), and they were fasted for 12 hours before the experiment. Then, equimolar amounts of ferritin, insulin, and insulin-ferritin nanoparticles dissolved in PBS were injected subcutaneously into these rats. Tail blood samples were collected before dosing and at 5, 15, 30, 60 minutes after dosing, and every hour thereafter. The experiment lasted for 12 hours, and the One-touch blood glucose meter ( Perfoma) was used to measure the blood glucose concentration of samples.

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Abstract

The invention discloses a nano-drug delivery system used for sustained release of subcutaneous insulin. The drug delivery system comprises ferritin. The construction of insulin-ferritin sustained release drug delivery systems is based on self-assembly mediated by the hydrophilic and hydrophobic interaction of the ferritin with the insulin. Meanwhile, the internal cavity of the ferritin may be loaded with drugs such as insulin secretagogues agents, metformin, alpha-glycosidase inhibitors, thiazolidinedione derivatives and DPP-4 enzyme inhibitors. The novel protein nano-drug carrier has the advantages of low immunogenicity, biodegradability, high insulin and anti-blood glucose drug loading capacity, capability of obviously controlling blood glucose after one-time administration, capability of realizing mass production, and low cost. The drug carrier and the application thereof in diabetes have great social benefit and economic benefit, and also have good application prospect.

Description

Technical field: [0001] The invention belongs to the intersecting fields of nanobiology, bionics and biomedicine. In particular, the present invention discloses a nanometer drug delivery system that can load insulin and realize slow release of insulin subcutaneously, and the delivery system includes ferritin. Background technique [0002] Diabetes mellitus is a chronic comprehensive disease mainly caused by the disorder of glucose metabolism caused by the absolute or relative deficiency of insulin, or the decreased sensitivity of target cells to insulin. The occurrence of type 2 diabetes is the joint action of peripheral insulin resistance and β-cell dysfunction. the result of. Diabetes and its complications have become one of the most troublesome diseases in the world. According to the latest data released by the International Diabetes Federation, the number of adults with diabetes (20-79 years old) worldwide has reached 425 million in 2017, and it is estimated that by 20...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/69A61K47/42A61K38/28A61P3/10
CPCA61K38/28A61K47/42A61K47/6949A61P3/10
Inventor 柯天一范克龙
Owner CHINA SCI XINYUN BIOTECH BEIJING CO LTD
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