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Multi-arm polyamino acid (ester) grafted polyethyleneimine copolymer, preparation method and application in gene delivery

A polyethyleneimine copolymer and polyethyleneimine technology are applied to multi-arm polyamino acid (ester) grafted polyethyleneimine copolymer and preparation method and the application field in gene transfer, which can solve the PEI charge density Insufficient concentration and other problems, to achieve the effect of good biocompatibility, low cytotoxicity and high transfection efficiency

Inactive Publication Date: 2009-11-11
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The synthesized multi-arm polyamino acid (ester) grafted polyethyleneimine copolymer has a branched structure and a concentrated charge density, which overcomes the high toxicity and non-degradability of the PEI homopolymer and the insufficient concentration of the linear polymer-grafted PEI charge density. shortcoming

Method used

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  • Multi-arm polyamino acid (ester) grafted polyethyleneimine copolymer, preparation method and application in gene delivery
  • Multi-arm polyamino acid (ester) grafted polyethyleneimine copolymer, preparation method and application in gene delivery
  • Multi-arm polyamino acid (ester) grafted polyethyleneimine copolymer, preparation method and application in gene delivery

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Synthesis of multi-armed poly-L-glutamate γ-benzyl esters using different polyamine initiators.

[0036] Under anhydrous and oxygen-free conditions, weigh 5 grams of γ-benzyl-L-glutamic acid-N-carboxylic acid anhydride and add them to different dry reaction ampoules, fill with nitrogen protection, and then add 500ml of anhydrous Chloroform, stir to make it dissolve; With the branched polyethyleneimine (branched PEI, MW=600; 1800) that molecular weight is 600 or 1800 as polyamine initiator, dissolve and prepare in anhydrous chloroform with the concentration of 0.01mmol / ml According to the volume of the initiator in Table 1, absorb and add to the corresponding reaction ampoule respectively, and stir the reaction at 30°C for 72 hours under temperature control; after the reaction is completed, settle in 5000ml of ether respectively, filter and dry cake, the white solid product multi-arm poly-L-glutamic acid γ-benzyl ester (multi-arm PBLG) was obtained, and the mo...

Embodiment 2

[0039] Example 2: Synthesis of multi-armed poly-L-aspartic acid β-benzyl esters using different polyamine initiators.

[0040] Under anhydrous and oxygen-free conditions, weigh 4.7 grams of β-benzyl-L-aspartic acid-N-carboxylic acid anhydride and add them to different dry reaction ampoules, fill with nitrogen protection, and then add 300ml of anhydrous Water N'N'-dimethylformamide, stirred to dissolve; branched polyethyleneimine (branched PEI, MW=600; 1800) with a molecular weight of 600 or 1800 was used as a polyamine initiator, and 0.01 mmol / ml concentration dissolved in anhydrous chloroform, according to the volume of the initiator in Table 2, were absorbed and added to the corresponding reaction ampoule, temperature control at 30 ° C stirring reaction for 72 hours; after the completion of the reaction, respectively Settled in 3000ml of ether, filtered and dried the filter cake to obtain the product multi-arm poly-L-aspartic acid β-benzyl ester (multi-arm PBLA). The produc...

Embodiment 3

[0043] Embodiment 3: Synthesis of multi-arm poly-L-glutamic acid (ester) graft polyethyleneimine copolymer by catalytic aminolysis method

[0044] Weigh respectively 2.2 grams (containing 10mmol benzyl ester group) of multi-arm poly-L-glutamic acid γ-benzyl ester (multi-arm PBLG 1 or multi-arm PBLG 3) in different reaction ampoules, and then according to Table 3 Add the corresponding polyethyleneimine (PEI) and 2-hydroxypyridine (HP) respectively in the molar amount of feeding, and the types of feeding PEI are respectively: linear polyethyleneimine with a molecular weight of 423 (linear PEI, MW=423), The branched polyethyleneimine (branched PEI, MW=600) that molecular weight is 600 or the branched polyethyleneimine (branched PEI, MW=1800) that molecular weight is 1800, see table 3; Dimethyl sulfoxide was stirred to dissolve it, the temperature was controlled at 38°C, and the reaction was stirred for 48 hours; The liquid after dialysis is lyophilized to obtain multi-arm poly-L...

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Abstract

The invention relates to a multi-arm polyamino acid (ester) grafted polyethyleneimine copolymer, a preparation method and application in gene delivery. The method includes the steps of: dissolving polyamine initiator in organic solvent and initiating the ring opening polymerization of alpha-amino acid-N-carboxylic acid anhydride protected by phenmethyl under anhydrous and oxygen free conditions to obtain the multi-arm polyamino acid (ester); and then causing the full or partial aminolysis reaction to occur between benzyl ester at the lateral chain of the multi-arm polyamino acid (ester) and amino group of polyethyleneimine to form the grafted copolymer. The polymer is a novel high-efficiency polycation gene vector, integrates the properties of polyethyleneimine and polyamino acid and is high in transfection efficiency, and the highest transfection efficiency to the mediate luciferase plasmid of Chinese Hamster Ovary epithelial cell is ten times than that of the Lipofectamine2000 of the commercial transfection reagent in US Invitrogen biomax under the same conditions; cytotoxicity is small; and cell survival rate is over 80% within the best transfection rate range, thus having broad application prospect.

Description

technical field [0001] The invention relates to multi-arm polyamino acid (ester) grafted polyethyleneimine copolymer, its preparation method and its application in gene transmission. Background technique [0002] With the development of modern biotechnology and the continuous improvement of the human gene pool, it has gradually become possible for humans to understand the root causes of certain diseases at the molecular level, providing a theoretical basis for gene therapy. In practice, how to obtain safe and effective gene carriers has increasingly become a bottleneck problem restricting the clinical application of gene therapy. The use of viral vectors to mediate gene transfer is the most widely used method in gene therapy, including retroviruses, adenoviruses, adeno-associated viruses, herpes simplex virus, vaccinia virus and other vectors. However, there are great safety hazards in the clinical application of viral vectors, which caused the first patient death in the hi...

Claims

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Application Information

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IPC IPC(8): C08G81/00C08G73/04C08G69/08C12N15/87C12N15/85
Inventor 陈学思陈磊田华雨夏加亮李非凡景遐斌
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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