Guanidinylation SS-PAAs polymer as well as preparation and application thereof

A technology of polyaminoamine and guanidinylation, which is applied in the direction of gene therapy, genetic material composition, and the use of vectors to introduce foreign genetic material, etc., to achieve the effect of improving transfection efficiency and reducing cytotoxicity

Inactive Publication Date: 2014-03-12
SHENYANG PHARMA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Guanidinylation SS-PAAs polymer as well as preparation and application thereof
  • Guanidinylation SS-PAAs polymer as well as preparation and application thereof
  • Guanidinylation SS-PAAs polymer as well as preparation and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0063] Example 1: Preparation of SS-PAAs with Agmatine Sulfate as Guanylation Reagent

[0064] Materials and Methods

[0065] 2,2,4,6,7-Pentamethyldihydrobenzofuran-5-sulfonyl chloride (Pbf-Cl): purchased from Jill Biochemical (Shanghai) Co., Ltd. N,N-bis(acryloyl)cystamine (CBA): purchased from Alfa Aesar (Tianjin) Chemical Co., Ltd. Agmatine sulfate: purchased from Changzhou Archie Biotechnology Co., Ltd. Trifluoroacetic acid (TFA): purchased from Tianjin Kemiou Chemical Reagent Co., Ltd. pcDNA3.1-EGFP-cDNA plasmid and plasmid extraction kit: purchased from Tiangen Biochemical Technology Co., Ltd. pSliencer TM 4.1 CMV-FANCF-shRNA plasmid, FANCF-shRNA, and 328-microRNA fragment: were designed, synthesized and purchased by Shanghai Bioengineering Co., Ltd. Lipofectamine2000: purchased from Invitrogen Company; ExGen500: purchased from Fermentas Company; MTT assay cell viability kit, cell experiment consumables, culture medium, agarose, etc.: all purchased from Baoxin Biolo...

Embodiment 2

[0073] Implementation example 2: prepare SS-PAAs with chlorhexidine acetate monomer as guanidinylation reagent

[0074] (1) Protection of Pbf-Cl on the guanidinium structure in chlorhexidine acetate

[0075] Put 1g of chlorhexidine acetate in a 50ml three-neck bottle, add 15ml of water to dissolve, cool down to below 0°C, and adjust the pH to 11; slowly add 2g of Pbf-Cl dissolved in 10ml of acetone dropwise, and the temperature is maintained at 0°C Below, the pH is maintained in the range of 10-13; after the dropwise addition, react below 0°C for 1 hour and at room temperature for 3 hours until the pH does not change. The progress of the reaction was monitored by TLC. Suction filtration, rotary evaporation to remove acetone in the filtrate, collect the acetone phase product, oven dry at 40-60°C overnight, weigh, and obtain chlorhexidine whose guanidine structure is protected by Pbf-Cl.

[0076] (2) Synthesis of guanidinated SS-PAAs polymer with chlorhexidine as monomer

[0...

Embodiment 3

[0081] Implementation example 3: Preparation of SS-PAAs with guanidine hydrochloride monomer as guanidinylation reagent

[0082] (1) Protection of Pbf-Cl on the guanidinium structure in guanidine hydrochloride

[0083] Put 1g of guanidine hydrochloride in a 100ml three-neck flask, add 5ml of water to dissolve, cool down to below 0°C, adjust the pH to 11; slowly add 3.02g of Pbf-Cl dissolved in 10ml of acetone dropwise, and maintain the temperature at 0°C Below, the pH is maintained in the range of 10-13; after the dropwise addition, react below 0°C for 1 hour and at room temperature for 1 hour until the pH does not change. The progress of the reaction was monitored by TLC. Suction filtration, collect the precipitated product, dry in an oven at 40-60°C overnight, and weigh to obtain guanidine hydrochloride whose guanidine structure is protected by Pbf-Cl.

[0084] (2) Synthesis of guanidinated SS-PAAs polymer with guanidine hydrochloride as monomer

[0085] Take 0.19g of the...

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Abstract

The invention belongs to the technical field of medicine, and relates to a guanidinylation SS-PAAs gene vector polymer as well as preparation and application thereof. The preparation method comprises the following steps: the guanidinylation SS-PAAs gene vector polymer is finally prepared from a cross-linking agent with a dual-acroloyl structure and a guanidinylation reagent through the processes of protection reaction performed on sulfonyl chlorides by sulfonyl chlorides, Michael addition polymerization and removal of protection of a benzene sulfonyl chloride guanidyl protecting agent in sequence. The polymer gene vector can be self-assembled with different kinds of gene segments to form a compound, so that the excellent biological membrane permeability is achieved, and the responsive degradation is realized in the reducible environment of cells, and the nuclear localization effect is achieved. The capability of loading the gene segments of the vector and the transport process can be regulated through controlling the guanidinylation reagent species, the proportion of the guanidinylation reagent in the polymer, and the molecular mass. The guanidinylation SS-PAAs gene vector can improve the transfection efficiency of genes, and reduce cytotoxicity, and is a novel gene vector adopted in the process of gene therapy.

Description

technical field [0001] The invention belongs to the field of new auxiliary materials for pharmaceutical preparations and new dosage forms of biological preparations, and relates to guanidinated polyaminoamine polymers containing disulfide bonds and their preparation and application. The carrier contains disulfide bonds and has good membrane permeability and cell Homeostasis reduction sensitivity, and peptidomimetic nuclear localization effects. Background technique [0002] As an emerging tumor treatment method, gene therapy has been widely favored by experts in the field of medicine in the past 20 years and has become an important part of the research and development of anti-tumor drugs. Gene therapy technology has entered the growth stage from the introduction period, and has the following obvious advantages: (1) It can treat the causes of disease occurrence and development, and is expected to achieve a fundamental cure for the disease; (2) Based on the principle of comple...

Claims

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Application Information

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IPC IPC(8): C08G73/00C12N15/87C12N15/85A61K48/00
Inventor 丁平田余涧坤
Owner SHENYANG PHARMA UNIVERSITY
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