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Dendritic human cell-penetrating peptide hPP7K and production method thereof, and hPP7K mediated plasmid DNA transfection method

A transmembrane peptide, human-derived technology, applied in the field of biomedicine, can solve the problems of insecurity and low transfection efficiency, and achieve the effects of low cost, low immunogenicity, and easy quality control.

Active Publication Date: 2017-10-13
肽泽(武汉)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a dendritic human-derived cell-penetrating peptide, its production and a method for mediating plasmid DNA transfection, so as to overcome the low transfection efficiency and potential unsafe factors of existing penetrating peptides lack of

Method used

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  • Dendritic human cell-penetrating peptide hPP7K and production method thereof, and hPP7K mediated plasmid DNA transfection method
  • Dendritic human cell-penetrating peptide hPP7K and production method thereof, and hPP7K mediated plasmid DNA transfection method
  • Dendritic human cell-penetrating peptide hPP7K and production method thereof, and hPP7K mediated plasmid DNA transfection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048](1) Weigh a certain amount of Fmoc-Arg(PBF)-Wang resin or H-Arg(PBF)-2cl resin, pour it into a glass reactor, add an appropriate amount of dichloromethane (DCM) to soak until the resin swells, and pump Dry;

[0049] (2) Add an appropriate amount of deprotection solution to the reactor, agitate with nitrogen for 0.5-2 hours, drain, add an appropriate amount of dimethylformamide (DMF) to the reactor, agitate with nitrogen, drain, repeat operation 1-10 Second-rate;

[0050] (3) Weigh the amino acid in each step equivalent to 1 times the molar amount of the resin, 1 times the molar amount of the carbodisubtype condensing agent (such as dicyclohexylcarbodiimide (DCC)) or benzotriazolium salt Type condensing agent (such as O-benzotriazole-tetramethyluronium hexafluorophosphate (HBTU) or O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboron Hydrochloric acid (TBTU)) was dissolved in dimethylformamide (DMF) or DCM or tetrahydrofuran (THF) and added to the reactor, reac...

Embodiment 2

[0058] (1) Weigh a certain amount of Fmoc-Arg(PBF)-Wang resin or H-Arg(PBF)-2cl resin, pour it into a glass reactor, add an appropriate amount of dichloromethane (DCM) to soak until the resin swells, and pump Dry;

[0059] (2) Add an appropriate amount of deprotection solution to the reactor, agitate with nitrogen for 0.5-2 hours, drain, add an appropriate amount of DMF into the reactor, agitate with nitrogen, drain, and repeat the operation 1-10 times;

[0060] (3) Weigh amino acids equivalent to 10 times the molar amount of the resin in each step, 10 times the molar amount of the carbon disubtype condensing agent (such as dicyclohexylcarbodiimide (DCC)) or benzotriazolium salt Type condensing agent (such as O-benzotriazole-tetramethyluronium hexafluorophosphate (HBTU) or O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboron Hydrochloric acid (TBTU)) was dissolved in dimethylformamide (DMF) or DCM or tetrahydrofuran (THF) and added to the reactor, reacted for 1-4h, a...

Embodiment 3

[0068] (1) Weigh a certain amount of Fmoc-Arg(PBF)-Wang resin or H-Arg(PBF)-2cl resin, pour it into a glass reactor, add an appropriate amount of dichloromethane (DCM) to soak until the resin swells, and pump Dry;

[0069] (2) Add an appropriate amount of deprotection solution to the reactor, agitate with nitrogen for 0.5-2 hours, drain, add an appropriate amount of DMF into the reactor, agitate with nitrogen, drain, and repeat the operation 1-10 times;

[0070] (3) Weigh the amino acid that is equivalent to 5 times the molar amount of the resin, the carbodisubtype condensing agent (such as dicyclohexylcarbodiimide (DCC)) or the benzotriazolium salt of 5 times the molar amount Type condensing agent (such as O-benzotriazole-tetramethyluronium hexafluorophosphate (HBTU) or O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboron Hydrochloric acid (TBTU)) was dissolved in dimethylformamide (DMF) or DCM or tetrahydrofuran (THF) and added to the reactor, reacted for 1-4h, and...

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Abstract

The present invention relates to the field of biomedicine, and discloses a novel dendritic human cell-penetrating peptide hPP7K and a production method thereof, and an hPP7K mediated plasmid DNA transfection method. According to the present invention, hPP7K is a novel dendritic human cell-penetrating peptide, is covalently or non-covalently linked to a marker or vector molecule at the C or N terminal, and carries a marker or cargo molecule to penetrate through the cell membrane so as to enter the cell; the cell-penetrating peptide hPP7K belongs to the improvement and modification of the human cell-penetrating peptide hPP7K, has characteristics of low immunogenicity, safety, low toxicity and high cell-penetrating effect, wherein the efficiency is higher than hPP10; the cell-penetrating peptide hPP7K is subjected to solid phase synthesis, such that the cost is low, and the quality control is easy to achieve; and the cell-penetrating peptide hPP7K can carry markers or cargo molecules to enter a variety of cells in the transmembrane manner, such that the cell-penetrating peptide hPP7K is the transmembrane transport carrier having the development prospects.

Description

technical field [0001] The invention relates to the field of biomedicine, and relates to a human-derived cell-penetrating peptide, its production and a method for mediating plasmid DNA transfection. Background technique [0002] At this stage, most diseases need to be diagnosed and treated at the molecular level, but facing the cell membrane, many macromolecular drugs with in vitro biological activity cannot enter the cell, so they cannot play their due role. Various techniques have been developed to overcome the transport barriers of macromolecules in vivo. Using viral vectors, although the transfer efficiency is high, its potential toxicity cannot be ignored. Non-viral methods, such as electroporation and microinjection, are traumatic to cells and may damage or even destroy cell membranes. Non-invasive methods such as liposomes require the destabilization of the membrane in a low-acid environment to release the drug into the cytoplasm, which has certain limitations for t...

Claims

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Application Information

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IPC IPC(8): C07K14/47C07K1/06C07K1/04C12N15/87
CPCC07K14/47C12N15/87
Inventor 刘岩松郭晓霞唐伟柳项段茹谢艳
Owner 肽泽(武汉)生物科技有限公司
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