Polypeptide hydrogel with slow-release exosomes and preparation method and application thereof

A hydrogel and exosome technology, applied in the fields of peptides, liquid delivery, pharmaceutical formulations, etc., can solve problems such as short half-life, and achieve the effect of slowing release, prolonging half-life, and having good clinical application prospects.

Active Publication Date: 2019-09-13
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although Exos has shown great potential for the treatment of AKI, its half-life in the blood circulation system after injection into the body is very short, and it is easily cleared by macrophages in the body. Therefore, it is necessary to design a slow-release carrier to deliver Exos to enhance its curative effect

Method used

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  • Polypeptide hydrogel with slow-release exosomes and preparation method and application thereof
  • Polypeptide hydrogel with slow-release exosomes and preparation method and application thereof
  • Polypeptide hydrogel with slow-release exosomes and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Example 1 Preparation of Exosomes derived from MSCs of the present invention

[0034] The preparation method is as follows:

[0035] 1. MSCs primary culture: Mice were killed by neck dislocation, soaked in 70% alcohol for 10 minutes for disinfection, the bilateral tibiae were separated, the bone marrow was washed with serum-free medium, the washing liquid was collected, centrifuged and resuspended, and MSCs were obtained by the adherent method.

[0036] 2. Culture medium collection: MSCs were cultured in DMEM medium (Gbico, USA) supplemented with 10% exosome-depleted fetal bovine serum (Exosome-depleted FBS, SBI, USA), and the culture supernatant was collected regularly.

[0037] 3. Centrifugal extraction: extract Exos by multiple centrifugation methods, centrifuge at 2000g×10 minutes to remove cell debris, centrifuge at 10000g×30 minutes to remove microvesicles (MVs), and centrifuge at 100000g×60 minutes to obtain Exos.

[0038] 4. Purification: The Exos obtained in t...

Embodiment 2

[0039] Embodiment 2 The preparation of polypeptide hydrogel of the present invention

[0040] According to the treatment requirements of the present invention, the hydrogel sustained-release Exos is used, and the sustained-release system is determined as follows according to the preliminary experimental results (therapeutic effect), experimental animals (C57 mice) and application sites (mouse kidney capsule):

[0041] Element Concentration (mg / ml) Volume (μl) Exosomes (Exos) 8 10 Self-assembled short peptide KMP2 10 13 sterile water 10 3

2 total 25

[0042] (1) The self-assembled short peptide KMP2 (KLDLPVGLIGKLDL) was synthesized and purified by Shanghai Botai Biotechnology. The dry powder was dissolved in sterile water to make a 10mg / ml stock solution, and 13μl was added to a 1.5ml EP tube;

[0043] (2) Add 10 μl of Exos derived from mesenchymal stem cells with a concentration of 8 mg / ml into the EP tube, mix well and set asi...

Embodiment 3

[0048] Embodiment 3 Preparation of polypeptide hydrogel of the present invention

[0049] (1) The self-assembling short peptide KMP2 (KLDLPVGLIGKLDL) was synthesized and purified by Shanghai Botai Biotechnology. Dissolving 300ug self-assembling peptide in 20μl water to make self-assembling peptide aqueous solution; dissolving 100ug exosomes in 15μl water to make exosomes Aqueous solution; add to EP tube;

[0050](2) Add 5 μl of water to the self-assembling peptide aqueous solution and exosome aqueous solution obtained in step (1) to make a total of 40 μl, and mix evenly to obtain a polypeptide hydrogel.

[0051] Prove beneficial effect of the present invention below by specific pharmacodynamics experiment:

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Abstract

The invention discloses a polypeptide hydrogel. The polypeptide hydrogel is prepared by mixing a self-assembled peptide (SAP) aqueous solution with exosomes (Exos). The present invention also discloses a preparation method and application of the polypeptide hydrogel. The polypeptide hydrogel provided by the invention can effectively slow down the release of the Exos and prolong the half-life of the Exos; and the released Exos have the activity similar to that of newly-separated Exos, and can effectively reduce HK2 cell injury caused by hypoxia and promote the recovery of vascular endothelial damage. The polypeptide hydrogel disclosed by the invention is capable of effectively reducing acute kidney injury and slowing down renal fibrosis by reducing kidney cell apoptosis and inflammatory factors, the expression amount of kidney injury marker protein-neutrophil gelatinase-related apolipoprotein (NGAL), and the synthesis and secretion of renal extracellular matrix (ECM), and has a good clinical application prospect.

Description

technical field [0001] The invention relates to the field of biological materials, in particular to an exosome slow-release polypeptide hydrogel that promotes the repair of damaged kidneys. Background technique [0002] Acute kidney injury (acute kidney injury, AKI) is a common clinical emergency characterized by a rapid decline in renal function, eventually leading to acute renal failure and even failure of other organs. There are many triggering factors of AKI, such as ischemia / reperfusion, application of surgical contrast medium, inappropriate medication, rhabdomyolysis, infection, etc. In recent years, the incidence of AKI has been increasing, and the incidence of AKI in hospitalized patients has reached 1%-5%, and it is increasing rapidly. At present, in the course of clinical treatment, there is still a lack of effective drugs for the prevention and treatment of AKI. After the occurrence of AKI, although the patient's renal function has been improved by conventional t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K9/06A61P13/12
CPCC07K7/08A61K35/28A61K47/42A61K9/06A61P13/12
Inventor 刘敬平周毅洁程惊秋陆燕蓉陈又南
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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