Enzyme-sensitive supramolecular hydrogel nano material, gelator and preparation method of gelator

A technology of supramolecular hydrogel and nanomaterials, which is applied in the field of supramolecular hydrogel and gel factor and its preparation, can solve the problems of difficult control and complicated preparation process, and achieve easy preparation, simple synthesis, and gelation conditions easily controlled effects

Active Publication Date: 2013-04-10
UNIV OF SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to propose a class of enzyme-sensitive supramolecular hydrogel nanomaterial gel factor and preparation method thereof, to obtain enzyme-sensitive supramolecular hydrogel nanomaterials, to overcome existing related supramolecular hydrogel materials The preparation process is complex and difficult to control, so it can be further used to detect specific enzyme activities related to apoptosis and assist cell survival in cell culture at the level of living cells

Method used

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  • Enzyme-sensitive supramolecular hydrogel nano material, gelator and preparation method of gelator
  • Enzyme-sensitive supramolecular hydrogel nano material, gelator and preparation method of gelator
  • Enzyme-sensitive supramolecular hydrogel nano material, gelator and preparation method of gelator

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Embodiment 1

[0031] Glycine-glycine-glycine-aspartic acid-valine-glutamic acid-aspartic acid and glycine-glycine-glycine-valine-aspartic acid-glutamic acid were first synthesized by solid-phase peptide synthesis -Aspartic acid two oligopeptide fragments were synthesized separately.

[0032] The first pure compound Y in this example 1 (Fmoc-EDA-Gly-Gly-Gly-Gly-Asp-Val-Glu-Asp-Acetyl) The synthetic route is as follows:

[0033]

[0034] The oligopeptide sequence glycine-glycine-glycine-aspartic acid-valine-glutamic acid-aspartic acid was first synthesized by solid-phase peptide synthesis method, according to: 1 mmol of 2-chlorotrityl chloride resin After swelling in 2-3 ml of N,N-dimethylformamide for 4-8 minutes, add 2 mmol of N-fluorenylmethoxycarbonyl-glycine, and then add 2 mmol of N,N-diisopropylethylamine , after 2-3 hours of reaction, react with 100 microliters of methanol for 5-10 minutes, cut off the fluorenyl moxycarbonyl protecting group of glycine, Kaiser test shows blue col...

Embodiment 2

[0046] Embodiment 2: In vitro enzyme activity detection experiment

[0047] In the in vitro experiment of this embodiment, the first pure compound Y with a concentration of 0.28 mmol per liter was used. 1 , in a concentration of 50 millimoles per liter of 4-hydroxyethylpiperazineethanesulfonic acid, 0.1% of 3-[(3-cholesterylaminopropyl) dimethylamino]-1-propanesulfonic acid, 50 milliliters Molar sodium chloride per liter, 10 mmol per liter of ethylenediaminetetraacetic acid and 5 caspase 3 with a unit volume of 50 microliters in a total volume of 60 microliters, incubated at 37 degrees Celsius Enzyme recognition and cleavage were completed in 100 minutes; the second pure compound Y was used at a concentration of 0.28 mmol per liter 2 , in a concentration of 50 millimoles per liter of 4-hydroxyethylpiperazineethanesulfonic acid, 0.1% of 3-[(3-cholesterylaminopropyl)dimethylamino]-1-propanesulfonic acid, 50 milliliters Molar sodium chloride per liter, 10 mmol per liter of ethy...

Embodiment 3

[0050] Embodiment 3: cell biocompatibility experiment

[0051] Firstly, the culture of human hepatoma cell HepG2 was carried out: at a volume concentration of 5% CO 2 The human liver cancer cell line HepG2 was cultured in a 37°C incubator in an air environment using DMEM medium containing 10% bovine serum albumin by volume; the cells in the logarithmic growth phase were washed three times with 0.01 moles per liter of sterile PBS buffer , and then digested with trypsin at a mass volume concentration of 0.25%; the sub-disc cells were rinsed with DMEM, then counted, and the cell concentration was diluted to 3×10 per ml 4 cells.

[0052] Then use human liver cancer cell HepG2 seed plate: plant HepG2 cells into 96-well microwell plate, 3×10 per well 3 cells in a volume concentration of 5% CO 2 DMEM medium containing 10% bovine serum albumin by volume was used for 24 hours in a 37°C incubator in an air environment.

[0053] Test the first pure compound Y separately 1 and a seco...

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Abstract

The invention discloses an enzyme-sensitive supramolecular hydrogel nano material which is easy to synthesize, a gelator and a preparation method of the gelator. The enzyme-sensitive supramolecular hydrogel nano material is characterized in that the gelator is synthesized by simple solid-phase polypeptide synthesis and conventional liquid-phase condensation reaction; gelatinization and gel dissolution are controlled in a manner of regulating pH; and meanwhile, the gelator comprises an oligopeptide sequence for identifying specificity of caspase 3, and thus, the gelator can be cut by the caspase 3 generated in the cell apoptosis process and survival time of cells can be prolonged. Compared with a conventional supramolecular hydrogel nano material, the enzyme-sensitive supramolecular hydrogel nano material disclosed by the invention has the advantages that complex reaction conditions are not required, the gelator is simple to synthesize and the gelatinization condition is easy to control; and the enzyme-sensitive supramolecular hydrogel nano material is sensitive to the specific caspase 3 generated in the cell apoptosis process, has excellent biocompatibility for cells and can be used for detecting activity of a specific enzyme related to apoptosis on the living cell plane and assisting cells to live in the cell culture.

Description

technical field [0001] The invention belongs to the technical field of supramolecular hydrogel nanomaterials, and in particular relates to an enzyme-sensitive supramolecular hydrogel and gel factor that can be used to assist cell survival and a preparation method thereof. Background technique [0002] Supramolecular hydrogels with oligopeptides protected by fluorenyl moxycarbonyl as gelling factors have been published in the Journal of the American Chemical Society (J.Am.Chem.Soc., 2009, 131, 11286-11287) According to the report of the phosphatase-regulated gelation process, the technology integrates biological processes, but has not further tested the biological application research of this material at the level of living cells, and there are no relevant reports on biocompatibility and degradability. . The application of caspase 3 in the degradation process of supramolecular hydrogels was reported in the journal "Lab Chip" (Lab Chip, 2010, 10, 1946-1951) of the Royal Socie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K1/16C07K1/06C07K1/04C12N5/00
CPCY02P20/55
Inventor 梁高林唐安明
Owner UNIV OF SCI & TECH OF CHINA
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