Tight-junction protein inhibitor polypeptide and application thereof

A connexin and inhibitor technology, applied in the field of relapsed and metastatic peptides, to improve the survival rate

Inactive Publication Date: 2015-06-10
SUZHOU PULUODA BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no well-developed tight junction protein inhibitor polypeptide, which is used to prevent and treat cancer recurrence and metastasis after breast cancer surgery

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Role of the tight junction protein inhibitor polypeptide on the migration of breast cancer cell MCF-7.

[0016] Using scratch test. First use a marker pen on the back of the 24-well plate, and use a ruler to draw horizontal lines evenly, about every 0.5~1cm, across the holes. Three lines were passed through each well; MCF-7 cells grown in logarithmic 5 Added to 24-well culture plate, cultivated for 24h. On the second day, compare the ruler with a 10 μl pipette tip, scratch perpendicular to the horizontal line on the back, and use the intersection of the scratch and the horizontal line on the back as a fixed observation point; add different concentrations of experimental wells and positive drug control wells respectively. The drug tight junction protein inhibitor polypeptide (Shanghai Sangong Synthetic) and the positive control drug vincristine; the same volume of solvent was added to the blank group, and five replicate holes were set in each well; put in 37 ° C, 5% CO...

Embodiment 2

[0018] Claudin inhibitor polypeptide has IC50 on the growth and survival of human breast cancer cells cultured in vitro.

[0019] Using the MTT colorimetric method. Human breast cancer cells growing logarithmically at 1.0×10 5 Put it into a 96-well culture plate and incubate for 24 hours. Add different concentrations of the experimental drug tight junction protein inhibitor polypeptide (Shanghai Sangon Synthetic) and the positive control drug paclitaxel to the experimental wells and positive drug control wells; add the same volume of solvent to the blank group. Set up five duplicate wells per well, culture for 48h, add MTT to each well at 0h, 2h, 8h, 14h, 20h, 24h, 36h, and 48h respectively, add DMSO after 4h, incubate for 30min, and incubate at 620nm in a microplate reader. The absorbance A value was measured, according to the formula human breast cancer cell growth inhibition rate=(1-absorbance value of the experimental group / absorbance value of the control group)×100%. Th...

Embodiment 3

[0021] In vivo activity of tight junction inhibitor polypeptides was tested in tumor models.

[0022] The MCF-7 tumor model was established, and the positive control drug paclitaxel was added; the same volume of solvent was added to the blank group, and three doses of polypeptide (Shanghai Sangong Synthetic) were set in the experimental group: 2, 4, and 8 μM / Kg. After 21 days, the number of surviving mice was observed, and the survival rate was calculated. The results showed that the tight junction protein inhibitor polypeptide can effectively protect mice and improve the survival rate of tumor-bearing mice, with a survival rate of 76.4%.

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Abstract

The invention relates to the field of medicine, in particular to a polypeptide capable of inhibiting tight-junction protein and preventing and treating a cancer relapse and transfer of the postoperative breast cancer. The brand new sequence of the polypeptide is LNLSSTLASGKDATGSYPTLMV. The polypeptide can inhibit proliferation and migration of breast cancer cells in an in-vitro mode, the survival rate of tumor-bearing mice is increased in an in-vivo test, and the polypeptide has the potential new medicine development value.

Description

technical field [0001] The invention relates to a tight junction protein inhibitor polypeptide and its application, in particular to a polypeptide capable of inhibiting tight junction protein, preventing and treating cancer recurrence and metastasis after breast cancer surgery. Background technique [0002] The female mammary gland is composed of skin, fibrous tissue, mammary glands and fat. Breast cancer is a malignant tumor that occurs in the mammary gland epithelial tissue. 99% of breast cancers occur in women and only 1% in men. At present, breast cancer has become a common tumor that threatens women's physical and mental health. [0003] The mammary gland is not an important organ to maintain human life activities, and breast cancer in situ is not fatal; but because breast cancer cells lose the characteristics of normal cells, the connections between cells are loose and easy to fall off. Once the cancer cells fall off, the free cancer cells can spread throughout the b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/00A61K38/16A61P35/00
CPCA61K38/00C07K14/00
Inventor 罗瑞雪
Owner SUZHOU PULUODA BIOLOGICAL SCI & TECH
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