Active peptide for promoting osteoblast proliferation and application thereof

A technology of osteoblasts and active peptides, applied in the field of active peptides, can solve the problems of no single purified active peptide products, unclear product ingredients, etc.

Active Publication Date: 2019-12-31
TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The preparation methods of bioactive peptides include acid method, alkali method and enzymatic method, among which the preparation of bioactive peptides by enzymatic method is more common. At present, many active peptides derived from bone collagen on the market are mostly enzymatic hydrolysis products of bone collagen, and there is no The active peptide product after single purification, the product composition is not clear, and various problems will be encountered in the later stage of drug and health product development process

Method used

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  • Active peptide for promoting osteoblast proliferation and application thereof
  • Active peptide for promoting osteoblast proliferation and application thereof
  • Active peptide for promoting osteoblast proliferation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Preparation of active peptide

[0038] According to the contents disclosed in Chinese patent application CN 105586379 A "a method for preparing collagen active peptides with the effect of inhibiting the proliferation of cancer cells", the ultrafiltration component CP1 is obtained, that is, the steps of the preparation method disclosed in the patent application are completed ( 1)-(8).

[0039] Reversed-phase silica gel column chromatography: Dissolve the ultrafiltration component CP1 sample in 1mL ultrapure water to prepare a 10mg / ml solution. Centrifuge and filter with 0.22μm disposable filter membrane. The bioactive peptides were separated and purified using an ODS-AQ reversed-phase silica gel column (50μm, 2.5mm×100mm), with a loading volume of 1mL. The eluent is: ultrapure water (liquid A) and chromatographic ethanol (liquid B). The flow rate is: 5mL / min. The elution step gradient is: 0-10min, 100%A; 10-30min, 96%A; 30-50min, 90%A; 50-70min, 87%A; 70-75min, ...

Embodiment 2

[0048] Based on the amino acid sequences of the five active peptides identified in Example 1, experimental samples were obtained by solid-phase synthesis to verify the proliferation activity of osteoblasts. Solid-phase synthetic peptide company: Gil Biochemical (Shanghai) Co., Ltd.

[0049] The result is Picture 11 As shown, when the concentration is 0.05mg / ml, compared with the control group, the five polypeptides have statistical differences (P <0.05), indicating that these five active peptides can significantly promote the proliferation of osteoblasts. However, this is not the case at 0.0125 mg / ml or 0.2 mg / ml, indicating that 0.05 mg / ml is the most suitable concentration of polypeptide to promote the proliferation of osteoblasts.

[0050] Cell proliferation detection

[0051] Firstly, 750 mice MT3T3-E1 per well were inoculated into three 96-well plates. After the cells were attached for 24 hours, the culture medium was aspirated and the active peptides selected at different con...

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Abstract

The invention discloses active peptides for promoting osteoblast proliferation and application thereof. The active peptides have amino acid sequences as shown in SEQ ID NO: 1 to SEQ ID NO: 5. The active peptides can promote osteoblast proliferation so as to prevent or treat osteoporosis. These active peptides can further be developed as pharmaceuticals, food or health products.

Description

Technical field [0001] The invention relates to the technical field of active peptides. More specifically, it relates to an active peptide that promotes the proliferation of osteoblasts and its application. Background technique [0002] Senile osteoporosis (OP) is a chronic and progressive disease characterized by decreased bone mass and bone microstructure and prone to fractures. As the age increases, the probability of suffering from osteoporosis will greatly increase. According to WHO statistics, there are approximately 75 million OP patients in the United States, Europe and Japan. The domestic situation is also not optimistic. A large-scale national epidemiological survey found that the prevalence of people over 60 has increased significantly. However, most of the drugs currently on the market for treating osteoporosis have large side effects and average effects. [0003] Biologically active peptides refer to peptide compounds that are beneficial to the life activities of b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/78C12N15/12A23L33/18A61K38/39A61P19/10
CPCA23V2002/00A61K38/00A23L33/18A61P19/10C07K14/78A23V2200/306A23V2250/55
Inventor 郭燕川王佳宁刘俊丽王富荣刘芳
Owner TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI
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