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Nano vesicle capable of co-transporting drugs and genes, manufacturing method and applications thereof

A nanovesicle and drug technology, applied in the fields of polymer chemistry and biomedical engineering, can solve the problems of wide particle size dispersion, difficult to control nanoparticle size, unfavorable in vivo application, etc., and achieve good stability and good biocompatibility. to achieve the effect of passive targeted aggregation

Inactive Publication Date: 2014-08-20
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, researchers mostly use cationic polymers to transfer genes. The cationic polymer chains combine with the phosphate skeleton of genes through electrostatic interaction to form nanoparticles. Aggregation and precipitation are not conducive to in vivo application

Method used

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  • Nano vesicle capable of co-transporting drugs and genes, manufacturing method and applications thereof
  • Nano vesicle capable of co-transporting drugs and genes, manufacturing method and applications thereof
  • Nano vesicle capable of co-transporting drugs and genes, manufacturing method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 PEI-PAsp (DIP / MEA) Preparation of nanovesicles

[0052] 1. Preparation of polymer PBLA:

[0053] The polymer is obtained through ring-opening polymerization of BLA-NCA with n-butylamine as an initiator. Add 2.49 g BLA-NCA (0.01 mol) to a 100 mL reaction flask, add 6 mL DMF to dissolve, and then add 60 mL anhydrous CH 2 Cl 2 , shake well. After that, 20 μL of n-butylamine (0.0002 mol) was added, shaken well, sealed, and stirred at 35 oC for 72 h. After the reaction, the reaction solution was dropped into excess cold ether (such as 500 mL) to precipitate, and the final product was obtained by suction filtration, repeated washing with anhydrous ether, and vacuum drying;

[0054] 2. Preparation of polymer PBLA-COOH:

[0055] Weigh 2.06 g Ba-PBLA (0.2 mmol) into a 100 mL three-neck flask, dry it in vacuum at 80 oC for 5 h, add 20 mL of freshly distilled chloroform (CHCl 3 ), after it dissolves, add 0.2 g of succinic anhydride (2 mmol, ~10 eq.), and stir a...

Embodiment 2

[0064] Example 2 Preparation of drug-loaded gene nanovesicles

[0065] Dissolve 1 mg 5-FC and 10 mg PEI-PAsp (DIP / MEA) in 1 mL DMSO, drop into 10 mL pH 7.4 hydrochloric acid buffer solution under the action of ice-bath ultrasound, and cross-link with oxygen for two hours, The prepared vesicle solution was dialyzed in water with pH 7.4 for three days to remove DMSO, filtered through a 450 nm filter membrane to remove large aggregates, and ultrafiltered with a molecular weight cut-off of 10 KDa to remove a small amount of residual 5- FC, to obtain nanovesicles loaded with 5-FC;

[0066] According to the molar ratio of the amino group in the polymer to the phosphate group in the plasmid DNA (N / P ratio) is 10:1, mix the CD gene with the PEI-PAsp(DIP / MEA) vesicle solution loaded with 5-FC and shake it After 15 s, it was left at room temperature for 30 min to obtain nanovesicles loaded with 5-FC and CD simultaneously.

[0067] The morphology of the resulting drug-loaded-gene nan...

Embodiment 3

[0068] Example 3 Determination of Drug Loading Efficiency and Encapsulation Efficiency of Nanovesicles

[0069] The standard curve of absorbance versus concentration (Y=0.000149+0.04347X, R=0.99994) was calculated by measuring the UV spectra of a series of known concentrations of 5-FC solutions, and the lyophilized load samples were dissolved in a pH value of 5 In PBS buffer solution, the absorbance of the sample solution at 275 nm was measured by a UV-visible spectrophotometer. Substituting the absorbance value into the standard curve equation to calculate the 5-FC concentration, thus obtaining the content of 5-FC in the sample, and obtaining the dry product mass according to the total mass of the freeze-dried sample container and the freeze-dried sample, thereby calculating the drug loading ratio and Encapsulation rate, the calculation formula is as follows:

[0070] Drug loading = content of 5-FC in vesicles / mass of vesicles × 100%

[0071] Encapsulation efficiency = 5-...

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Abstract

The invention discloses a nano vesicle capable of co-transporting drugs and genes, a manufacturing method and applications thereof. The component of the nano vesicle is polyethyleneimine-b-polyaspartic acid (diisoprylamino ethylamine / cysteine). The preparation method comprises the following steps: using n-butylamine to induce aspartate benzyl-N-carboxyl anhydride to carry out ring-opening polymerization reactions so as to obtain poly(aspartate benzyl), subjecting the terminal amino groups to carboxylation reactions, then mixing the reaction products with diisoprylamino ethylamine and cysteine to carry out ammonolysis reactions so as to obtain polyethyleneimine-b-polyaspartic acid (diisoprylamino ethylamine / cysteine) with carboxylated terminals, and finally subjecting the polyethyleneimine-b-polyaspartic acid (diisoprylamino ethylamine / cysteine) with carboxylated terminals to amidation reactions so as to obtain the polyethyleneimine-b-polyaspartic acid (diisoprylamino ethylamine / cysteine). The nano vesicle can be taken as the co-transportation carrier for hydrophilic drugs and genes, the hydrophobic layer of the vesicle has an acid-response property, at the same time, the disulfide bond, which is used for the binding and crosslinking of hydrophobic layer, has a reduction sensitivity, and the outer layer of the vesicle namely the polyethyleneimine layer has a proton-buffering effect, so the vesicle can release the genes and drugs intelligently.

Description

technical field [0001] The invention belongs to the fields of macromolecular chemistry and biomedical engineering, and in particular relates to a nanovesicle which can be used as a dual-carrier material for transmitting new drugs and genes, a manufacturing method and an application thereof. Background technique [0002] Malignant tumors seriously threaten the survival and health of human beings, and the treatment of cancer is one of the most challenging topics at present. At present, surgical treatment is mostly used clinically, but this is usually applicable to the situation where the tumor has not started to spread to the whole body. The application of radiotherapy and chemotherapy has significantly improved the therapeutic effect of malignant tumors. Among them, chemotherapy has considerable therapeutic potential for various hematological malignancies, blastoma, etc., but the toxic and side effects of chemotherapy drugs on normal tissues and organs is the main problem th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/34A61K48/00A61K9/51A61P35/00
Inventor 蒋庆王露苑园园曹众帅心涛
Owner SUN YAT SEN UNIV
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