Natural killer T cell activator polypeptide and application thereof

A natural killer, activator technology, applied in the direction of peptides, peptide/protein components, medical preparations containing active ingredients, etc., can solve the problems that have not yet been developed, and achieve the effect of increasing the survival rate

Active Publication Date: 2014-09-10
深圳市旷逸生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no developed natural killer T cell activator polypeptide drug for the treatment of acute lymphoblastic leukemia

Method used

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  • Natural killer T cell activator polypeptide and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Chemical Synthesis of Peptides

[0016] The peptides were synthesized using Fmoc chemistry. The synthesis reaction proceeds from the C-terminus to the N-terminus, and the Rink medium (available from Advanced ChemTech) has free amino groups, and the amino acids are connected sequentially. During each ligation step, the amino acid residues are activated, and the activation mixture contains 4 times as many HBTU, HOBt, DIEA and Fmoc-amino acids as there are free amino groups on the medium. After each amino acid linking reaction, a mixture of pyridine / acetic acid / N-methylimidazole (4:1:0.5) was used to block unlinked free amino groups for 10 min. After each amino acid ligation reaction and before the next amino acid is ligated, the Fmoc-group on the medium must be removed, and the Fmoc-group is removed using dimethylformamide containing 20% ​​piperidine, which takes 15 minutes. Finally, after all amino acid residues are linked sequentially, the peptide is cleaved from the ...

Embodiment 2

[0019] IC50 values ​​of natural killer T cell activator matrix metalloproteinase-2 in vitro.

[0020] Matrix metalloproteinase-2 is activated by cleaving the fluorescent substrate Mca-Arg-Pro-Lys-Pro-Val-Glu-Nval-Trp-Arg-Lys(Dnp)-NH 2 to detect (excitation wavelength=320 nm, emission wavelength=405 ​​nm). Reactions were performed in 100 μl reactions at 37°C. At the beginning of the reaction, 4 μl of natural killer T cell activator 2 stock solution (1 ng / μl) was added to the reaction system, and the final concentration of the substrate was 10 μM. The calculated IC50 value was 2.05 μM.

Embodiment 3

[0022] In Vivo Viability of Natural Killer T Cell Activators Using an Endotoxic Shock Model

[0023] Before establishing the endotoxic shock model, we first determined the LD50 of LPS (E. coli 0111: B4, purchased from sigma company) in mice, which was 50 μg per mouse. In the experiment, we used 100 μg per mouse, In this way, all the mice in the control group died. In our previous scientific research work, we found that Regasepin2 (another peptide inhibitor, published) can improve the survival rate of C57BL / 6 mice during endotoxin shock. In order to establish endotoxin shock model in our mice, we did a positive control experiment with Regasepin2. The mice in the control group were injected with 100 μg of LPS, while the mice in the Regasepin2 experimental group were injected with 0.7 mg of polypeptide 5 minutes after LPS injection. By making a Kaplan-Meier survival curve, it was found that Regasepin2 can effectively protect mice injected with 100 μg and improve the survival ra...

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PUM

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Abstract

The invention relates to the field of drugs and particularly relates to a polypeptide capable of activating natural killer T cells, inhibiting matrix metalloproteinases-2 and relieving destruction of acute inflammatory reaction to an organism. The polypeptide has the sequence as follows: RQQQEATRPRTAAANGLMQT which is a brand new sequence. The polypeptide can be used for inhibiting the activity of matrix metalloproteinases-2 in vitro and increasing the survival rate of endotoxin shock rats in an in-vivo test so as to have a potential novel drug development value.

Description

technical field [0001] The invention relates to a natural killer T cell activator and application thereof, in particular to a polypeptide capable of activating natural killer T cells and treating acute lymphocytic leukemia. Background technique [0002] Acute lymphoblastic leukemia (ALL) is a progressive malignant disease characterized by large numbers of immature white blood cells that resemble lymphoblasts. These cells can be found in the blood, bone marrow, lymph nodes, spleen and other organs. Acute lymphoblastic leukemia accounts for 80% of acute leukemia in children, and the peak incidence is between 3 and 7 years old. ALL can also occur in adults and accounts for 20% of all adult leukemias. In recent years, with the deepening of medical research, great progress has been made in the understanding and treatment of acute lymphoblastic leukemia. Biological targeted therapy for tumors is very promising. Targeted therapy is to block a certain process of tumorigenesis to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K38/10A61P35/02
Inventor 罗瑞雪
Owner 深圳市旷逸生物科技有限公司
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