Anti-tumor peptide sourced from hemocyanin of litopenaeus vannamei and application thereof

A technology for tumors and short peptides, which is applied in the field of tumor-suppressing short peptides and their coding genes and applications. It can solve the problems of long peptide chains, difficult chemical synthesis, and difficult purification, and achieve high anti-tumor activity.

Inactive Publication Date: 2017-08-11
SHANTOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This kind of anti-tumor peptide directly derived from the body generally has a long peptide chain, which is difficult to synthesize by chemical methods, difficult to purify, and has the disadvantages of unstable structure and easy inactivation.

Method used

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  • Anti-tumor peptide sourced from hemocyanin of litopenaeus vannamei and application thereof
  • Anti-tumor peptide sourced from hemocyanin of litopenaeus vannamei and application thereof
  • Anti-tumor peptide sourced from hemocyanin of litopenaeus vannamei and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Solid-phase synthesis, cleavage, purification and identification of tumor-suppressing short peptide B1

[0027] The present invention utilizes a polypeptide synthesizer to synthesize the short peptide for inhibiting tumors by a solid-phase synthesis method:

[0028] 1) With DMF as the solvent, the concentration of various α-amino acids protected by Fmoc is 0.25M, the concentration of HBTU solution and HOBt solution is 0.33M, the concentration of piperidine solution is 200ml / L, and the concentration of DIEA solution is 174.2ml / L.

[0029] 2) Weigh 0.05mmol of Fmoc-Ala-Wang resin (functional group content 0.33mmol / g) into a solid-phase reactor, add 8ml of DCM to swell overnight, and remove the solvent under reduced pressure. Add 8ml of piperidine solution with a concentration of 200ml / L, react at room temperature for 5min, and drain; then add 8ml of piperidine solution with a concentration of 200ml / L, react for 20min at room temperature, and drain; wash with 8...

Embodiment 2

[0038] Example 2: Determination of Antitumor Activity of Short Peptide B1 for Inhibiting Tumors

[0039] (1) Use human cervical cancer cells (HeLa) in logarithmic phase of growth, digest and resuspend the cells to obtain a cell suspension, and dilute the cells to 50,000 cells / mL. Add 100 µL to each well of the 96-well plate, put the paved 96-well plate in the incubator, and store at 37°C, 5% CO 2 Incubate overnight (>8 hours) under conditions.

[0040] (2) Use 0.01M pH7.4 PBS to dissolve the short peptide at 1 mg / mL. Dilute the short peptide sample to 50 µg / mL with complete medium. Discard the cell culture medium cultivated overnight, add 100 µL of 50 µg / mL polypeptide to each well, and repeat 3 times in 4 parallels to each well. At the same time, an equal volume of 50 μg / ml 5-FU and 0.01M pH7.4 PBS were used as positive and negative controls, at 5%37 o C conditions were cultured for 18 h.

[0041] (3) Next, discard the culture medium and dry it, add 50 μL of ice-cold 500...

Embodiment 3

[0046] Example 3: Comparison of the anti-tumor activity of the tumor-suppressing short peptide B1 and other short peptide fragments in hemocyanin

[0047] (1) Synthesize 7 different hemocyanin short peptide fragments and name them respectively (B1, B2, B3, B13, B14, S8, S9,). The short peptide of the present invention is B1.

[0048] (2) Use human cervical cancer cells (HeLa) in the logarithmic phase of growth, digest and resuspend the cells to obtain a cell suspension, and dilute the cells to 50,000 cells / mL. Add 100 µL to each well of the 96-well plate, put the paved 96-well plate in the incubator, and store at 37°C, 5% CO 2 Incubate overnight (>8 hours) under conditions.

[0049] (3) Use 0.01M pH7.4 PBS to dissolve the short peptide at 1mg / mL. Dilute different short peptide samples to 50 µg / mL with complete medium. Discard the cell culture medium cultivated overnight, add 100 µL of 50 µg / mL polypeptide to each well, and repeat 3 times in 4 parallels to each well. At the...

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Abstract

The invention discloses anti-tumor oligopeptide B1 sourced from hemocyanin of litopenaeus vannamei. The oligopeptide has an amino acid sequence as shown in SEQ ID NO:1. The oligopeptide can be used for inhibiting growth of cervical cancer cells HeLa, and can become an excellent anti-cancer medicine with low side effect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a tumor-suppressing short peptide, its coding gene and its application. Background technique [0002] Cancer has high morbidity and mortality rates worldwide. In recent decades, human beings have made unremitting efforts to treat cancer, and the research on cancer mainly focuses on developing new treatment methods and reducing the side effects of treatment methods on patients. The commonly used cancer treatment methods are mainly surgery or chemotherapy, but in many cases not only cannot treat patients efficiently, but also have a high risk of complications. Some cancer treatment drugs not only have multiple side effects, but also may cause cause multidrug resistance. [0003] As a new type of drug for treating cancer, antitumor peptides are attracting more and more attention because they are selective for tumor cells and not easy to cause drug resistance. [0004] Many antitumor ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/795A61K38/41A61P35/00
CPCA61K38/00C07K14/795
Inventor 章跃陵刘尚杰王帆黄河郑莉媛
Owner SHANTOU UNIV
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