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A Novel Penetrating Peptide Mediating Drug Delivery and Its Application

A technology of drugs and anti-tumor drugs, which is applied in the field of membrane-penetrating peptides and the application of the membrane-penetrating peptides in drug delivery and tumor treatment. It can solve the problems of low introduction efficiency, difficulty in promotion, death, etc. best therapeutic effect

Active Publication Date: 2021-10-29
北京翼方生物科技有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are common problems such as low import efficiency, difficulty in popularization, and easy to cause cell damage or even death.
All in all, there is still a lack of an efficient and feasible method for direct introduction of therapeutic drugs

Method used

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  • A Novel Penetrating Peptide Mediating Drug Delivery and Its Application
  • A Novel Penetrating Peptide Mediating Drug Delivery and Its Application
  • A Novel Penetrating Peptide Mediating Drug Delivery and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Synthesis of Penetrating Peptides

[0043] 1) Activation of resin: Weigh 1000mg of wang resin, add 10-15ml (submerge the whole resin) in DMF and soak for 30min to make it fully swell;

[0044] 2) Deprotection: Soak the DMF of the resin to remove it by pressure filtration, add 10ml of DMF solution containing 20% ​​piperidine, react with nitrogen blowing and boiling for 15min, then remove it by pressure filtration, remove the FMOC group of the amino group, and wash the resin with 10ml of isopropanol Three times, then wash three times with 10ml DMF, and then use the ninhydrin method to detect that the resin should turn black or purple;

[0045] 3) Condensation reaction: connect the next amino acid, weigh the amount of Fmoc-amino acid to be 1.4mmol / g resin, 910mg TBTU, add 10ml DMF and 0.45g HOBt and mix evenly, add 0.52ml DIEA to make the reaction solution, nitrogen at room temperature Boiling reaction 2h. After the reaction was complete, the resin was washed ...

Embodiment 2

[0050] Embodiment 2: Cytotoxicity experiment

[0051] 1) Take a 96-well plate and add 7×10 3 1 cell, 37 ° C, 5% carbon dioxide incubator for 24 hours, so that the cells adhere to the wall;

[0052] 2) Prepare culture medium containing different concentrations of the penetrating peptide of SEQ ID No.3, and use the medium without the penetrating peptide as the negative control well (Control), and incubate at 37° C. with 5% carbon dioxide for 1-5 hours;

[0053] 3) Add 20 μl MTT to each well of adherent cells, continue to incubate for 24 hours, discard the culture medium, add 150 μl DMSO (dimethyl sulfoxide) to each well, and shake for 10 minutes;

[0054] 4) Select a wavelength of 490nm or 570nm, and calculate the cell survival rate by the light absorption value on the microplate reader immunodetector. The results are shown in Table 1. The results show that the multiple penetrating peptides of the invention have no obvious cytotoxicity. Select the result of SEQ ID No: 3 as s...

Embodiment 3

[0057] Example 3: Membrane-penetrating peptide and fluorescence detection experiment

[0058] 1) Add the culture medium containing 100 μM concentration of FITC fluorescence-labeled SEQ ID No.2 membrane-penetrating peptide to the adherent cervical cancer (Hela) cells, incubate for 30min-4h, carefully suck off the medium, and wash the adherent cells three times with PBS ;

[0059] 2) Add fixative, 10min-20min, fix the cells and wash the cells 2-3 times with PBS;

[0060] 3) Add DAPI staining solution to stain for 5min-10min, then wash with PBS 2-3 times;

[0061] 4) Observe with a fluorescence microscope under the excitation light of 488nm and 543nm respectively, the results are shown in figure 2 . The results showed that the membrane-penetrating peptide successfully penetrated the membrane and delivered the fluorescent marker into the cell.

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Abstract

The invention discloses a drug-loaded membrane-penetrating peptide, clarifies the structure of the polypeptide, and provides partial data on how the polypeptide efficiently mediates drug delivery into cells. The invention has broad application prospects in the preparation of anti-tumor therapeutic drugs.

Description

technical field [0001] The present invention relates to the field of biotechnology, more specifically, to a membrane-penetrating peptide and the application of the membrane-penetrating peptide in drug delivery and tumor treatment. Background technique [0002] According to the statistics of the World Health Organization in 2017, there are more than 14 million new cancer cases in the world every year, causing 8.8 million deaths, accounting for nearly one-sixth of the total annual death toll in the world, and this number is still increasing. In 2010, the annual economic cost caused by cancer was 1.16 trillion US dollars, and cancer has become one of the most serious diseases that threaten human life, health and social development. Among the current treatment methods, chemical drug therapy (chemotherapy) is currently and will be used as one of the main means of clinical tumor treatment for a long time, and drug delivery is one of the key issues in chemotherapy. Due to the sele...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06A61K31/337A61P35/00
CPCA61K31/337A61K47/42C07K7/06
Inventor 姜海业
Owner 北京翼方生物科技有限责任公司
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