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Fusion protein, and pharmaceutical composition and use thereof

A technology of fusion protein and composition, applied in the field of fusion protein, can solve problems such as difficult to use and short half-life, and achieve the effect of promoting cartilage repair, long half-life, and promoting new bone formation

Active Publication Date: 2019-05-17
LINK HEALTH GRP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the half-life of AC-100 is relatively short, and its plasma half-life is only about 20 minutes, so it is difficult to be used in the treatment of diseases (such as osteoporosis) that require systemic administration. Drug frequency, it is possible to meet the urgent clinical needs in the field of orthopedic disease treatment, and has broad application prospects

Method used

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  • Fusion protein, and pharmaceutical composition and use thereof
  • Fusion protein, and pharmaceutical composition and use thereof
  • Fusion protein, and pharmaceutical composition and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Synthesis of Example 1 AC-100 Peptide

[0040] AC-100 peptide (SEQ ID NO: 1) was entrusted to Shanghai Aobo Biotechnology Co., Ltd. to synthesize by conventional solid-phase technology, and the purity of the synthesized peptide was >98%.

Embodiment 2

[0041] Construction and expression of embodiment 2 recombinant fusion protein strain

[0042] The construction of recombinant expression strains of fusion proteins HSA021-1 and HSA021-2 was entrusted to Nanjing GenScript Biotechnology Co., Ltd. The main steps are: synthesizing the target gene (SEQ ID NO: 2, corresponding to the nucleotide sequence encoding HSA021-1, which contains the sequence encoding the signal peptide; SEQ ID NO: 4, corresponding to the nucleotide sequence encoding HSA021-2 sequence, which contains the sequence encoding the signal peptide), was connected to the expression yeast expression vector pPICZaA through the same digestion by XhoI / NotI double digestion, and 10 μg of the expression vector after the connection was used to transfect Pichia pastoris (Pichia pastoris) X-33 , PCR identified positive clones. Selected positive clones were cultured in BMGY medium, OD 600 When it reached 3.0, the cells were harvested, resuspended in BMGY medium, and induced ...

Embodiment 3

[0044] Example 3 Effects on Bone Marrow Mesenchymal Stem Cells (BMSCs)

[0045] Three 8-week-old SD rats were killed by dislocation. After being sterilized with 70% ethanol, the abdominal cavity was cut open and the femur was separated. Use a syringe to draw low-sugar DMEM culture solution (containing 1 mL / L heparin) to flush the bone marrow cavity repeatedly to flush out the bone marrow.

[0046] The bone marrow fluid was made into a single-cell suspension, and DMEM-F12 complete medium containing 10% FBS (fetal bovine serum, fetal bovine serum) was added, and 10 7 The density of each / mL was inoculated in a culture flask, placed at 37°C, 5% CO 2 Cultivate in an incubator, change the culture medium after 48 hours, and then change the culture medium every 3 days to remove non-adherent hematopoietic cells. When the cells reach 80% to 90%, subculture, add the complete culture medium of mesenchymal stem cells to resuspend the cells, and store at 37°C, 5% CO 2 cultured in an incu...

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Abstract

The invention relates to a fusion protein, and a pharmaceutical composition and a use thereof, and belongs to the field of fusion protein. The fusion protein comprises 2 molecules of AC-100 linked tohuman serum albumin through a connexon, and the AC-100 has the amino acid sequence shown in SEQ ID NO:1. The 2 molecules of the AC-100 in the fusion protein show synergistic effect, have remarkable effect of promoting cartilage repair and new bone formation, and can be used for repairing cartilage and treating osteoarthritis, osteoporosis or bone fracture. At the same time, the fusion protein overcomes the shortcomings of instability and short half-life of AC-100 drugs, has significant longer half-life period in plasma and joint cavities, and is beneficial for reducing the frequency and dosageof drug administration.

Description

technical field [0001] The invention relates to a fusion protein, its pharmaceutical composition and application, and belongs to the field of fusion proteins. Background technique [0002] Osteoarthritis (OA) is a common joint disease characterized by joint pain and stiffness. Cartilage damage is the main cause of osteoarthritis. The disease occurs frequently after middle age, and women are more than men. The prevalence rate of the 40-year-old crowd is 10% to 17%, that of the over 60-year-old is 50%, and that of the over-75-year-old crowd is as high as 80%. The disease has a certain rate of disability. With the aging population, osteoarthritis will become an important issue affecting people's quality of life, and the market demand for osteoarthritis drugs will continue to expand. [0003] At present, clinically, the therapeutic drugs for osteoarthritis are divided into specific therapeutic drugs and non-specific therapeutic drugs. Non-specific therapeutic drugs are mainl...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61K38/18A61K47/62A61P19/02A61P19/00A61P19/10
Inventor 宋燕许元生张时群苏博雅
Owner LINK HEALTH GRP
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