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EphA7-Fc fusion protein and application thereof in drugs for preventing and/or treating osteoporosis diseases

An epha7-fc-ha, fusion protein technology, applied in the field of biomedicine, can solve problems such as drug side effects, and achieve the effects of alleviating bone loss, promoting osteoblasts, and inhibiting osteoclasts

Active Publication Date: 2021-06-18
SCI RES TRAINING CENT FOR CHINESE ASTRONAUTS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the current osteoporosis drugs have a certain therapeutic effect on osteoporosis, some drugs will produce certain side effects after long-term use, and new therapeutic targets for osteoporosis need to be further developed
At the same time, there is still a lack of bone formation drugs that stimulate the activity of osteoblasts, and the application of such drugs to people with slow bone loss is conducive to maintaining the integrity of the trabecular bone structure
There is no report on the relationship between Eph-A7 and osteoporosis

Method used

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  • EphA7-Fc fusion protein and application thereof in drugs for preventing and/or treating osteoporosis diseases
  • EphA7-Fc fusion protein and application thereof in drugs for preventing and/or treating osteoporosis diseases
  • EphA7-Fc fusion protein and application thereof in drugs for preventing and/or treating osteoporosis diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Effect of EphA7-Fc on bone function of osteoblasts:

[0050] 1. Using human osteoblast cell line MG63 cells, using 5.0 μg / mL bovine serum albumin (BSA) control group and 5.0 μg / mL EphA7-Fc and induction differentiation medium (containing 50 g / mL vitamin C, 2.0 mM β-sodium glycerophosphate, 10nM dexamethasone in DMEM high glucose complete medium) to induce osteotropic differentiation.

[0051] 2. After 2 days of differentiation induction, the cells were collected with TriZol and fixed with 4.0% paraformaldehyde.

[0052] 3. Alkaline phosphatase ALP staining method:

[0053] The cells transfected with siRNA were sucked out of the culture medium with a pipette, and the cell surface was fully washed 2-3 times by adding cold PBS, and the cells were fixed with 4% paraformaldehyde fixative solution for 10 minutes, then the paraformaldehyde fixative solution was removed, and the Wash twice with PBS, prepare ALP staining solution (VECTOR Blue SK-5300) according to the followin...

Embodiment 2

[0086] Effect of EphA7-Fc on bone function of osteoclasts:

[0087] 1. Isolate bone marrow mononuclear cells from the bone marrow of mice to induce osteoclast differentiation. C57BL / 6 mice were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., SPF grade. Male C57BL / 6 mice aged 6-8 weeks were killed by cervical dislocation, sterilized by 75% alcohol soaking; bilateral femurs and tibias were taken, and the bone marrow cavity was washed several times with sterile PBS drawn from a sterile syringe, and the liquid was collected in a 15mL centrifuge tube , centrifuged at 1000r / min for 5 minutes; discarded the supernatant, added 5mL erythrocyte lysate, and lysed for 5 minutes; centrifuged, discarded the supernatant, and used 5ml of α-MEM medium containing 10% fetal calf serum and double antibody (complete culture base)+10ng / mL M-CSF resuspended, inoculated in 10cm petri dish, placed in 5% CO 2 , cultured in a cell incubator at 37°C for 24 hours; collecte...

Embodiment 3

[0101] Effects of EphA7-Fc and bone-specific targeting EphA7-Fc-HA on bone loss in mice induced by ovariectomy:

[0102] 1. C57BL / 6 mice, purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., SPF grade, female 8 weeks old, bred to 4 months old, using sterilized surgical instruments to remove bilateral ovaries from the back of the mouse , to construct the ovary removal model, and the sham operation group retained the ovary;

[0103] 2. After the mice recovered for one week, the mice were randomly divided into 8-10 mice in each group. The sham operation group was injected with 100 μL PBS (phosphate buffered saline), and the operation control group was injected with 100 μL PBS. The four groups were injected with different doses of EphA7-Fc or bone-specific targeting EphA7-Fc-HA drugs, and the doses of the four groups were as follows (ratio of drug / mouse body weight): EphA7-Fc (25 μg, 1.0 mg / kg), EphA7-Fc -HA (10μg, 0.4mg / kg), EphA7-Fc-HA (25μg, 1.0mg / kg)...

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Abstract

The invention relates to an EphA7-Fc fusion protein and application thereof in drugs for preventing and / or treating osteoporosis diseases, and belongs to the technical field of biomedicine. Both the EphA7-Fc fusion protein and a bone targeting EphA7-Fc-HA fusion protein can obviously promote the functions of osteoblasts and obviously inhibit the functions of osteoclasts; and the EphA7-Fc fusion protein and the bone targeting EphA7-Fc-HA fusion protein can be used for relieving bone loss caused by different reasons.

Description

technical field [0001] The invention relates to an EphA7-Fc fusion protein and its application in drugs for preventing and / or treating osteoporosis, and belongs to the technical field of biomedicine. Background technique [0002] Osteoporosis, defined as a systemic metabolic bone disease, is an insidious, common, but underdiagnosed disease; characterized by decreased bone mass and microarchitectural disruption of bone, resulting in increased bone fragility and predisposition to Fractures, insufficient generation of bone organic components, secondary calcium salt reduction, and destruction of bone micropore structure are the main manifestations. Osteoporosis has become a worldwide problem, and it is another disease that seriously endangers human health after tumors and cardiovascular diseases. Studies have shown that the incidence of osteoporosis ranks seventh among common diseases and frequently-occurring diseases. At present, there is no safe and effective radical cure in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00A61K38/17A61K47/68A61P19/10
CPCC12N9/12C12Y207/10001A61K38/177A61K47/68A61P19/10C07K2319/30
Inventor 李英贤李玉恒厉建伟凌树宽
Owner SCI RES TRAINING CENT FOR CHINESE ASTRONAUTS
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