Selenium-containing teriparatide and fusion polypeptide for promoting osteoblast differentiation and application of selenium-containing teriparatide and fusion polypeptide

A technology of osteoblast differentiation and teriparatide, which is applied in the field of active peptides, can solve the problems that have not yet been determined, the nutritional and physiological roles of selenium and bone health are not clear, and growth retardation, so as to promote the mineralization of osteoblasts and alleviate the Pain, the effect of treating osteoporosis

Pending Publication Date: 2022-05-31
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although selenium deficiency causes growth retardation, it has not been established whether this growth inhibition is associated with changes in bone metabolism
At the same time, there are a variety of selenium-containing enzymes expressed in bone cells, but the nutritional and physiological roles of selenium and bone health are still unclear

Method used

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  • Selenium-containing teriparatide and fusion polypeptide for promoting osteoblast differentiation and application of selenium-containing teriparatide and fusion polypeptide
  • Selenium-containing teriparatide and fusion polypeptide for promoting osteoblast differentiation and application of selenium-containing teriparatide and fusion polypeptide
  • Selenium-containing teriparatide and fusion polypeptide for promoting osteoblast differentiation and application of selenium-containing teriparatide and fusion polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 The effect of selenium-containing teriparatide on the proliferation of mouse osteoblasts MC3T3-E1

[0032] based on Figure 1-Figure 3 The amino acid sequences of three kinds of teriparatide containing selenium were designed and obtained, and experimental samples were obtained by solid-phase synthesis method to verify the proliferation activity of osteoblasts. Solid-phase synthetic peptide company: Gill Biochemical (Shanghai) Co., Ltd.

[0033] Detection of proliferation rate of mouse osteoblast MC3T3-E1:

[0034] Select osteoblasts MT3T3-E1 in log phase growth, according to 2 × 10 4 Cells were added to a 96-well plate at a density of cells / mL, with 100 μL of cell suspension per well. The experiment was set to 5 groups. Blank control group: only DMEM medium containing 1% fetal bovine serum was added; positive control group: except for the addition of 1% fetal bovine serum. In addition to the DMEM medium containing 1% fetal bovine serum, teriparatide was add...

Embodiment 2

[0035] Example 2 The effect of selenium-containing teriparatide on alkaline phosphatase in mouse MT3T3-E1 cells

[0036] Detection of MT3T3-E1 alkaline phosphatase activity in mouse osteoblasts:

[0037] Select osteoblasts MT3T3-E1 in log phase growth, according to 2 × 10 4 Cells were added to a 96-well plate at a density of cells / mL, with 100 μL of cell suspension per well. The experiment was set to 5 groups. Blank control group: only DMEM medium containing 1% fetal bovine serum was added; positive control group: except for the addition of 1% fetal bovine serum. In addition to the DMEM medium containing 1% fetal bovine serum, teriparatide was added with final concentrations of 0; 62.5; 125; 250; 500ug / mL; No. 1 peptide group: except for the addition of DMEM containing 1% fetal bovine serum. In addition to the base, the final concentrations of 0; 62.5; 125; 250; 500ug / mL were added with No. 1 selenium-containing teriparatide; No. 2 peptide group: In addition to the addition o...

Embodiment 3

[0038] Example 3 The effect of selenium-containing teriparatide on mineralized nodules of mouse MT3T3-E1 cells

[0039] Determination of mineralized nodules in mouse osteoblasts MT3T3-E1:

[0040] Select osteoblasts MT3T3-E1 in log phase growth, according to 1 × 10 5 Add cells / mL to a 6-well plate, 2 mL of cell suspension per well, aspirate and discard the original medium after culturing for 24 hours, and replace it with a medium without fetal bovine serum. Culture medium without fetal bovine serum, the experiment was set to 5 groups, blank control group: only DMEM medium containing 1% fetal bovine serum and three osteogenic inducers (dexamethasone 10nmol, β-glycerophosphate sodium 10mmol) were added. , ascorbic acid 5 μg / ml); positive control group: except for the addition of DMEM medium containing 1% fetal bovine serum and three osteogenic inducers (dexamethasone 10 nmol, β-glycerophosphate 10 mmol, ascorbic acid 5 μg / ml), Teriparatide with a final concentration of 250ug / m...

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Abstract

The invention discloses selenium-containing teriparatide and fusion polypeptide for promoting osteoblast differentiation and application of the selenium-containing teriparatide and fusion polypeptide. The parent amino acid sequence of the selenium-containing teriparatide disclosed by the invention is as shown in SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNF. The selenium-containing teriparatide disclosed by the invention can be used for preparing selenium-containing teriparatide. The selenium-containing teriparatide provided by the invention can effectively promote osteoblast proliferation and mineralization, prevent and / or treat osteoporosis and relieve pain of osteoporosis patients, and the polypeptides have the characteristics of high efficiency and low toxicity, and can be further developed into drugs.

Description

[0001] technical background [0002] The present invention relates to the technical field of active peptides. More specifically, it relates to a selenium-containing teriparatide for promoting osteoblast differentiation and its application. Background technique [0003] In recent years, with the development of the aging population in our country, the incidence of osteoporosis (OP) and the total number of people with osteoporosis have gradually increased, which has become an increasingly important health problem for human beings, and its treatment cost is high. , many complications, poor quality of life of patients, bring a heavy burden to the family and society. How to effectively prevent and treat osteoporosis is the focus of current research and an important problem that needs to be solved urgently. Relevant studies have confirmed that osteoblasts play a key role in the development and pathological process of osteoporosis. They not only play an irreplaceable role in the f...

Claims

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Application Information

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IPC IPC(8): C07K14/635C07K19/00A61K38/29A61K47/64A61P19/10
CPCA61K38/29A61K47/643A61P19/10C07K14/635C07K14/765C07K14/77A61K38/00C07K2319/00Y02A50/30
Inventor 吴晖贺萍张猛猛赖富饶张丽娜
Owner SOUTH CHINA UNIV OF TECH
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