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Non-viral gene vector constructed on basis of N-terminal octadecane acylated antibacterial peptide

A gene carrier and antimicrobial peptide technology, applied in the field of non-viral peptide carriers, can solve the problems of low toxicity, etc., and achieve the effects of easy biodegradation, lower application cost, and simple design and synthesis methods

Inactive Publication Date: 2012-10-10
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the reverse melittin has similar antibacterial activity, but has lower toxicity to normal cells (J. Am. Chem. Soc., 2008, 118, 8989)

Method used

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  • Non-viral gene vector constructed on basis of N-terminal octadecane acylated antibacterial peptide
  • Non-viral gene vector constructed on basis of N-terminal octadecane acylated antibacterial peptide
  • Non-viral gene vector constructed on basis of N-terminal octadecane acylated antibacterial peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1: the synthesis of stearyl-Mel

[0046] a. Resin pretreatment: Add Rink-Amide-MBHA resin with an amino molar mass of 0.2 mmol into the reactor, add 10 mL of dichloromethane and stir for 30 min to fully swell the resin, and then drain the solvent under reduced pressure.

[0047] b. Deprotection of F-moc: add 10~15mL deprotection reagent into the reactor, stir for 2min, then drain, repeat 4 times to completely remove the Fmoc group, and finally wash with DMF to remove the deprotection reagent; deprotection The reagent is piperidine / DMF=1:4 (V / V).

[0048] c. Condensation: 0.6~0.8 mol Fmoc group-protected amino acid, N-hydroxybenzotriazole, O-benzotriazole-N,N,N',N'-tetramethylurea-hexa Dissolve fluorophosphate in 3~5mL DMF, then add 1.2~1.6 mol diisopropylethylamine and mix to obtain a mixed solution, then put it into the reactor, stir and react for 40~60min; the whole reaction process is protected by argon , the degree of reaction was detected by the indene...

Embodiment 2

[0054] Example 2: stearyl- r Synthesis of Mel

[0055] d. Extension of the peptide chain: Repeat steps b and c in the order of QQRKRKRKIWSILAPLGTTLVKLVAGIG until the peptide grafting is completed. Other steps are the same as in Example 1. The amino acid sequence of the obtained analogue is opposite to that of stearyl-Mel.

[0056] The molecular weight of the product prepared by the above method characterized by ESI-MS is shown in Table 1.

Embodiment 3

[0057] Embodiment 3: the synthesis of stearyl-Mel-1

[0058] d. Extension of the peptide chain: Repeat steps b and c in the order of GIGAVLKVLTTGLPALISWRRRRRRRR until the peptide grafting is completed. Other steps are the same as in Example 1. The sequence of the obtained analog was that the IKRKRQQ sequence at the end of stearyl-Mel was replaced by RRRRRRR.

[0059] The molecular weight of the product prepared by the above method characterized by ESI-MS is shown in Table 1.

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PUM

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Abstract

The invention discloses a non-viral gene vector constructed on the basis of N-terminal octadecane acylated antibacterial peptide. The vector utilizes the characteristics of melittin, i.e. endosome-destroying activity and membrane penetrating activity, moreover, a nuclear localization sequence and a polyarginine sequence are added into the melittin sequence, and then a series of forward and reverse analogues are synthesized, and octadecanoic acid is connected with the N terminals of the obtained analogues, so that the analogues and plasmid DNA (deoxyribonucleic acid) form stable nano-particles. An experiment result shows that because the non-viral gene vector constructed on the basis of N-terminal octadecane acylated antibacterial peptide has higher transfection efficiency and lower cytotoxicity and the polypeptide vector is easy to biodegrade, the non-viral gene vector is more suitable for application in vivo.

Description

technical field [0001] The invention relates to a class of non-viral peptide vectors that can be used for gene therapy, in particular to a class of non-viral gene vectors constructed on the basis of N-terminal 18 alkanoylated antimicrobial peptides. Background technique [0002] Tumor is one of the important diseases that threaten human health. At present, the main means of tumor treatment are surgery, radiotherapy and chemotherapy. Although these methods can treat or slow down the growth of tumors to a certain extent, they often produce certain toxic and side effects, cause certain damage to the human body, and seriously reduce the quality of life of patients. Therefore, the development of new therapeutic methods with high efficiency and low toxicity has become an urgent problem in the field of tumor treatment. With the development of genomics, gene therapy has ignited new hope for the cure of tumors (Nature, 1998, 392, 25). However, how to safely and efficiently transfe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C07K14/435
Inventor 王锐张伟宋竟婧张邦治
Owner LANZHOU UNIVERSITY
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