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Acid-sensitive degradation-controllable serial polymers with branched structures and preparation method thereof

A branched structure, polymer technology, applied in the fields of biomedicine and polymer medical materials, can solve problems such as limiting gene expression and limiting release

Inactive Publication Date: 2013-08-21
四川中科惠泽环保科技集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the strong electrostatic interaction also limits the release of genes from the complex after entering the cell fluid, limiting gene expression

Method used

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  • Acid-sensitive degradation-controllable serial polymers with branched structures and preparation method thereof
  • Acid-sensitive degradation-controllable serial polymers with branched structures and preparation method thereof
  • Acid-sensitive degradation-controllable serial polymers with branched structures and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1 has the preparation method of the acid-sensitive cationic polymer gene carrier of branched structure

[0021] Take 0.283 g of ethylene glycol diglycidyl ether (n=1) and place it in a double-necked flask, add 1 mL of DMF, stir and dissolve, then add 0.5 g of diamine-type acid-sensitive monomer (i.e., ethylene glycol diglycidyl ether) to it. Glycerol ether: diamine-type acid-sensitive monomer molar ratio = 1: 1) Stir evenly, and react for 48 hours under anhydrous and anaerobic room temperature conditions; Add 0.01% triethylamine) and dialyze for 24-48 hours; accurately measure 1 mL of the dialysis product to freeze-dry, weigh the mass after freeze-drying, and obtain the concentration of the polymer in 1 mL of the dialysis solution. The remainder is stored frozen in liquid form. The polymerization product is Polymer 1.

[0022] Take 0.424 g of ethylene glycol diglycidyl ether (n=1) and place it in a double-necked flask, add 1 mL of DMF, stir and dissolve, th...

Embodiment 21

[0025] Example 2 1 HNMR detection of the structures of polymers 1, 2, and 3

[0026] The structures of the synthesized polymers 1, 2, 3 were determined by NMR. Use BRUKERAVANCE III HD (400MHz) nuclear magnetic resonance instrument detection 1 H NMR. The solvent is CD 3 Cl.

Embodiment 3

[0027] Embodiment 3 measures the cytotoxicity of polymkeric substance with MTT method

[0028] NIH / 3T3 cells were seeded into 96-well plates (1×10 4 each well), add 200 μL of DMEM medium containing 10% fetal bovine serum to each well at 37°C, 5% CO 2 Conditioned for 24h. Discard the medium, add 180 μL of fresh DMEM medium to each well and 20 μL of 20 mM HEPES solution to prepare a series of cationic polymer solutions, so that the final concentration of the polymer is: 1 mg / mL, 5 mg / mL, 10 mg / mL, 50 mg / mL mL, 100mg / mL, 500mg / mL, 1000mg / mL, 5000mg / mL. 37°C, 5% CO 2 Conditions for another 24 h. Add 20 μL of MTT solution (5 mg / mL) to each well, and continue to incubate at 37° C. for 4 h. Terminate the culture, discard the culture supernatant in the well, add 100 μL dimethyl sulfoxide (DMSO), and shake gently for 10 min. Measure the absorbance value of each well at 570nm, and calculate the cell viability (see figure 2 ).

[0029] Such as figure 2It was found that the cyt...

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Abstract

The invention discloses a preparation method of acid-sensitive degradation-controllable serial polymers with branched structures. The polymers are synthesized from ethylene glycol diglycidyl ether A and a diamine acid sensitive orthoester monomer B serving as frameworks, the molar ratio of A:B is 1-2:1, wherein the structural formulae is shown in the specification. Open loops of epoxy groups at both ends of A are polymerized with amino groups at both ends of B. When the molar ratio of A and B is 1:1, the polymers exist mainly in linear forms. With increase of molar ratio of A, the linear polymers are crosslinked (branched), so that net shapes are formed. When the molar ratio of A:B reaches 2:1, the polymers form three-dimensional netty structures. With increase of molar ratio of A: B, the branching degree of the polymers is improved. The polymers have cations and have the characteristics of acid sensitivity, degradation controllability and good biocompatibility, so that the polymers have important research value and application prospect in the field of gene therapy.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and polymer medical materials, and specifically relates to a series of polymers with branched structure, acid sensitivity and controllable degradation and a preparation method thereof. Background technique [0002] Gene therapy refers to inserting genes that can produce specific proteins into patient cells to correct genetic defects or treat acquired diseases, including cancer, AIDS, diabetes, etc. It is a new treatment method that has emerged in the past two decades and has broad development prospects. A safe and effective gene delivery system is a major bottleneck restricting the implementation of gene therapy. The most studied cationic polymer carrier is polyethylimide (PEI). High transfection efficiency due to its proton sponge effect. PEI with a molecular weight lower than 1.8KDa has almost no transfection effect. With the increase of molecular weight and branching degree, its ability t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08G73/00C12N15/63A61K48/00A61K47/34
Inventor 王睿唐汝培巩凯
Owner 四川中科惠泽环保科技集团有限公司
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