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Application of regulatory peptide and mansonone E in preparation of improved medicament for treating leukemia and application of regulatory peptide and mansonone E

A technology for leukemia and mansonone, which is applied in the field of biopharmaceuticals to achieve the effects of moderate cost, convenient medication and small side effects

Active Publication Date: 2019-06-11
THE PEOPLES HOSPITAL SHAANXI PROV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The results of this study show that mansonone E can induce K562 cell apoptosis by activating caspase-8 and caspase-3 pathways; Bcl-2 family proteins are not involved in the regulation of apoptosis (see: Mansonone E on the proliferation and Apoptosis, Miao Yudi, et al., Chinese Herbal Medicine, Volume 40, No. 9), but after years of research, the inventor believes that the drug still has room for improvement

Method used

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  • Application of regulatory peptide and mansonone E in preparation of improved medicament for treating leukemia and application of regulatory peptide and mansonone E
  • Application of regulatory peptide and mansonone E in preparation of improved medicament for treating leukemia and application of regulatory peptide and mansonone E
  • Application of regulatory peptide and mansonone E in preparation of improved medicament for treating leukemia and application of regulatory peptide and mansonone E

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1 The cultivation of K562 leukemia cells

[0034]Tumor cell line: K562 leukemia cells (preserved by Hematology Laboratory of Shaanxi Provincial People's Hospital). After recovering from liquid nitrogen, K562 cells were placed in RPMI-1640 culture medium containing 10% inactivated fresh calf serum at 37°C. The volume fraction was 5% CO2, cultured under saturated humidity conditions, and the cell viability was kept >97%. The K562 cells (1X 105 / ml) in the exponential growth phase were divided into the control group and the experimental group for future use.

Embodiment 2

[0035] Example 2 Cell Proliferation Inhibition Experiment

[0036] Cell proliferation inhibition experiment: Add K562 cells in the logarithmic growth phase to 24-well plate at 1×105 / mL, 1 mL per well, the drugs were 10 μg / mL polypeptide group, 25 μg / mL mansone E drug group, 10 μg / mL polypeptide + 25 μg / mL manthonone E drug group, PBS blank control group, counted after 12, 24, 36, and 48 hours, counted live cells by trypan blue exclusion test, repeated the experiment 3 times, and took mean, to plot growth curves, see figure 1 Shown, wherein the polypeptide sequence is shown in SEQ ID NO: 1.

[0037] from figure 1 It can be seen that the inhibitory effect of peptides alone on cells is very weak, while the combination of peptides and mansonone E can significantly inhibit cell proliferation and promote cell apoptosis, and the cell survival rate is less than 5% at 48 hours. Exhibited an excellent inhibitory effect.

Embodiment 3

[0038] The expression measurement of embodiment 3 protein

[0039] Determination of protein expression by Western blotting: Take K562 cells in the logarithmic growth phase, centrifuge, prepare a cell suspension with a concentration of 3×105 / mL in RPMI 1640 medium containing 10% fetal bovine serum, and place in a 6-well plate Inoculate 1 mL per well, place the plate at 37°C, 5% CO 2 incubator. Add 10 μg / mL polypeptide group, 25 μg / mL mansonone E drug group, 10 μg / mL polypeptide+25 μg / mL mansonone E drug group, PBS blank control group, centrifuge after 0 and 48 hours after administration respectively Collect cells, lyse cells with protein lysate, centrifuge to remove cell debris, cook at 95°C for 10 min, quantify protein, 40 μg / well, and load on 10% SDS-polypropylene gel electrophoresis. After the protein on the gel was transferred to the PVDF membrane, it was blocked overnight in 5% skimmed milk, and the primary antibody was added according to the antibody dilution ratio to i...

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Abstract

The invention relates to a regulatory peptide and mansonone E in preparation of an improved medicament for treating leukemia and application of the regulatory peptide and the mansonone E. Through model screening, the regulatory peptide capable of promoting the activity of the mansonone E is obtained, the regulatory peptide and the mansonone E are combined to prepare the medicament for treating leukemia. The medicament has the advantages of being small in dosage, moderate in cost, convenient to take, small in side effect and the like.

Description

technical field [0001] The invention relates to the use of regulatory peptide and mansonone E in the preparation of improved leukemia medicine, belonging to the field of biopharmaceuticals. Background technique [0002] Leukemia is a malignant clonal disease of hematopoietic stem cells. Clonal leukemia cells proliferate and accumulate in bone marrow and other hematopoietic tissues due to mechanisms such as uncontrolled proliferation, differentiation disorder, and apoptosis inhibition, and infiltrate other non-hematopoietic tissues and organs, while inhibiting normal hematopoietic function. Clinically, different degrees of anemia, hemorrhage, infection, fever, enlargement of liver, spleen, lymph nodes and bone pain can be seen. According to reports, the incidence of leukemia in various regions of my country occupies the sixth place among various tumors. [0003] According to the differentiation degree of leukemia and the length of natural course, it can be divided into acut...

Claims

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Application Information

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IPC IPC(8): A61K38/16A61P35/02A61P35/04A61K31/352A61K31/282A61K33/243
Inventor 苗玉迪
Owner THE PEOPLES HOSPITAL SHAANXI PROV
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