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Cell penetrating peptide hPP-chol, production thereof, and cell penetrating peptide hPP-chol mediated plasmid DNA transfection method

A hpp-chol, membrane-penetrating peptide technology, which is applied in the preparation methods of peptides, other methods of inserting foreign genetic materials, recombinant DNA technology, etc., can solve the problems affecting the application prospects and the low efficiency of mediated plasmid DNA transfection, etc. achieve low immunogenicity

Active Publication Date: 2017-07-04
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

Especially human-derived penetrating peptide (hCPP), compared with other biological sources of CPPs, hCPP is less likely to cause the human body's immune response, and there are relatively few potential unsafe factors (such as hPP10 publication number: 102863516A) , but for hPP10, the efficiency of mediating plasmid DNA transfection is low, which affects its application prospect as a drug intracellular delivery vehicle

Method used

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  • Cell penetrating peptide hPP-chol, production thereof, and cell penetrating peptide hPP-chol mediated plasmid DNA transfection method
  • Cell penetrating peptide hPP-chol, production thereof, and cell penetrating peptide hPP-chol mediated plasmid DNA transfection method
  • Cell penetrating peptide hPP-chol, production thereof, and cell penetrating peptide hPP-chol mediated plasmid DNA transfection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] (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;

[0048] (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;

[0049] (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, reacted for 1-4h, an...

Embodiment 2

[0061] (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;

[0062] (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;

[0063] (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

[0075] (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;

[0076] (2) Add an appropriate amount of deprotection solution to the reactor, agitate it with nitrogen for 0.5-2 hours, drain it, add an appropriate amount of DMF into the reactor, agitate it with nitrogen, drain it, and repeat the operation 1-10 times;

[0077] (3) Weigh amino acids equivalent to 5 times the molar amount of the resin in each step, 5 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, reacted for...

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Abstract

The invention belongs to the field of biological medicine, and discloses production of a novel cholesterol-modified human-derived cell penetrating peptide hPP-chol, and a cell penetrating peptide hPP-chol mediated plasmid DNA transfection method. The sequence of the novel cholesterol-modified human-derived cell penetrating peptide hPP-chol is Chol-KIPLPRFKLKCIFCKKRRKR. The C terminal or the N terminal of the novel cholesterol-modified human-derived cell penetrating peptide hPP-chol is connected with a marker or a carried molecule via covalence connection or non-covalence connection; and the marker or the carried molecule is carried by the novel cholesterol-modified human-derived cell penetrating peptide hPP-chol to enter into cells via penetrating cell film. The novel cholesterol-modified human-derived cell penetrating peptide hPP-chol is obtained based on improvement and modification of human-derived cell penetrating peptide, is safe, and is low in immunogenicity and toxicity, is obtained via solid phase synthesis, is low in cost, is excellent in cell penetrating effect, and can be widely used in the fields of drug, health care product, skin care product, transfection reagent, and diagnostic reagent practical production; and quality control is convenient to realize.

Description

technical field [0001] The invention relates to a cell-penetrating peptide, a method for producing it and mediating plasmid DNA transfection. Background technique [0002] Cell membranes consist of lipid bilayers with an interior of hydrophobic, nonpolar molecules. The cell membrane is a barrier for substances to enter and exit the cell, which only allows non-lipid-soluble molecules with a molecular weight of less than 600 Da to enter living cells. Although this barrier has a protective effect on the body, it also makes it difficult for some valuable hydrophilic macromolecular drugs to penetrate the cell membrane and enter the interior of the cell to reach an effective therapeutic concentration, making this type of therapeutic value but non-cell penetrating and easy to degrade The application of hydrophilic molecules in cell biology, pharmacy and other research fields is greatly limited. [0003] In the past few decades, it has been found that some peptides and proteins ca...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K1/06C07K1/04C12N15/87
CPCC07K7/08C12N15/87Y02P20/55
Inventor 刘岩松郭晓霞唐伟柳项段茹许力谢艳
Owner 肽泽(武汉)生物科技有限公司
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