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Tubulin polymerizing agent polypeptides 1 and application thereof

A tubulin and polymerizing agent technology, applied in the fields of peptide/protein components, peptides, drug combinations, etc., can solve the problem of lack of specificity of small molecular polymerizing agents, so as to increase the survival rate, improve the survival rate, and promote the polymerization activity. Effect

Inactive Publication Date: 2013-09-18
SUZHOU GULI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many successful small-molecule polymerizers can promote the activity of tubulin at the nanomolar level, but small-molecule polymerizers lack specificity. If the lack of specificity of tubulin polymerizers is used for a long time in chronic diseases, large side effect

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] The effect of tubulin polymerizer polypeptide 1 on the polymerization and depolymerization of tubulin in vitro.

[0017] Take fresh bovine brain tissue, peel off meninges and large blood vessels, cut into pieces, wash 1-2 times with cold MES buffer solution, add MES buffer solution at a ratio of 0.5-1ml per gram of brain tissue, at 4℃, use electric Homogenize in a homogenizer; centrifuge at 105000g for 1 hour at 4°C, take the supernatant, add an equal volume of microtubule polymerization buffer, and keep it in a water bath at 37°C for 30 minutes. Centrifuge at 105000g at 26°C for 1h, take the precipitate, add about 1 / 10 of the homogenized volume of cold MES buffer, stir gently or crush the precipitate with a homogenizer; put the suspension in an ice bath for 30 minutes to completely dissolve the precipitate. Use Lowry’s method to determine protein content (SDS polyamide gel electrophoresis). Tubulin was diluted to 4-5 mg / ml with MES buffer, and stored in liquid nitrogen. ...

Embodiment 2

[0021] Tubulin polymerizer polypeptide 1 has an IC50 for the growth and survival of tumor cells in vitro.

[0022] Using MTT colorimetric method. The logarithmic growth of U937 cells were added to a 96-well culture plate at 1.0×105, and cultured for 24 hours. Different concentrations of the experimental drug tubulin polymerizer polypeptide 1 and the positive control drug paclitaxel were added to the experimental wells and the positive drug control wells respectively; Add the same volume of solvent to the group. Set up five multiple wells in each well, culture for 48h, respectively at 0h, 2h, 8h, 14h, 20h, 24h, 36h, and 48h. Add MTT to each well. After 4h of action, add DMSO, incubate for 30min, in the microplate reader at 620nm Determine the value of absorbance A, according to the formula: tumor cell growth inhibition rate = (1-experimental group absorbance value / control group absorbance value)×100%. The IC50 of the experimental drug was calculated to be 15.43μM.

Embodiment 3

[0024] The tumor model was used to detect the in vivo activity of tubulin polymerizer polypeptide 1.

[0025] The U937 tumor model was established, the positive control drug paclitaxel; the blank group was added with the same volume of solvent, and the experimental group was set with 3 doses: 0.75, 1.5μM, 3μM mg / Kg. After 21 days, observe the number of surviving mice and calculate the survival rate. The results show that tubulin polymerizer polypeptide 1 can effectively protect mice and improve the survival rate of tumor-bearing mice, with a survival rate of 59.1%.

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PUM

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Abstract

The invention relates to the field of medicaments, and in particular relates to polypeptides capable of promoting polymerization of tubulin and treating acute lymphatic leukemia. The sequence of the polypeptides is SWAAGSYFWQG which is a brand-new sequence. The polypeptides can promote the polymerization of tubulin in vitro at a level of 1 micromole, and can improve the survival rate of tumor-bearing mice in an in vivo test, so that the polypeptides have a potential new medicament development value.

Description

Technical field [0001] The present invention involves microtubin polypeptides 1 and its application, which specifically involves polypeptides that have promoted microtubinal protein aggregation and the treatment of acute lymphocytic leukemia. Background technique [0002] Acute lymphocytic leukemia (ALL) is a kind of malignant disease, which is characterized by a large number of immature leukocytes similar to lymphocytes.These cells can be found in blood, bone marrow, lymph nodes, spleen and other organs.Acute lymphocytic leukemia accounts for 80%of children's acute leukemia, and the incidence is between 3 and 7 years old.ALL can also occur to adults, accounting for 20%of all adult leukemia.With the deepening of medical research in recent years, the understanding and treatment of acute lymphocytic leukemia have made great progress.Among them, microtubin plays an important role in acute lymphocytic leukemia. [0003] Micro tube is the main component of the cell skeleton, α -Morbin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06A61K38/08A61P35/02
Inventor 吴娟
Owner SUZHOU GULI BIOTECH
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