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Fusion polypeptide, application of fusion polypeptide in enhancement of stem cell homing ability and anti-apoptotic ability and pharmaceutical composition

A technology that fuses peptides and stem cells, applied in the field of medicine, can solve the problems of high application cost of chemokine SDF-1, poor survival ability of stem cells, poor targeting of stem cells, etc., to achieve enhanced targeting homing efficiency, good citation prospects, non-degradable effect

Pending Publication Date: 2020-02-18
中国人民解放军陆军特色医学中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although stem cell transplantation therapy, as a promising treatment, has been widely carried out in clinical practice and animal experiments, and has achieved good progress and gratifying curative effects, however, this technology faces two problems in the clinical application process. The big bottleneck problem: (1) stem cells have poor targeting, the number of cells colonized in the damaged area is small, and the utilization rate of stem cells is low; (2) the survival ability of stem cells reaching the damaged area after transplantation is poor, and a large number of rapid apoptosis occurs, which has been reported90 More than 100% of the cells undergo rapid and massive apoptosis within a few days after implantation, and less than 3% of the surviving cells after two weeks
However, the application cost of chemokine SDF-1 is high, and its short half-life (30 minutes) is easy to degrade, which brings inconvenience to the clinical application of SDF-1.

Method used

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  • Fusion polypeptide, application of fusion polypeptide in enhancement of stem cell homing ability and anti-apoptotic ability and pharmaceutical composition
  • Fusion polypeptide, application of fusion polypeptide in enhancement of stem cell homing ability and anti-apoptotic ability and pharmaceutical composition
  • Fusion polypeptide, application of fusion polypeptide in enhancement of stem cell homing ability and anti-apoptotic ability and pharmaceutical composition

Examples

Experimental program
Comparison scheme
Effect test

preparation example 1

[0072] Used to illustrate the synthesis of the fusion polypeptide provided by the invention

[0073] Entrusted Suzhou Qiangyao Biotechnology Co., Ltd. to synthesize SEQ ID NO.2 (represented by amino acids 1 to 17 of Nt of SDF-1, polypeptide B), SEQ ID NO.7 (polypeptide D, Nt of SDF-1 1st to 17th amino acid+GRGDS), SEQID NO.4 (polypeptide E, GRGDS).

preparation example 2

[0075] Used to illustrate the isolation and culture of mouse BMSC

[0076] 4-6 weeks old wild-type C57BL / 6 mice were sacrificed by cervical dislocation and soaked in 70% alcohol for 3 minutes. Under sterile conditions, the long bones (femur and tibia) of the lower limbs of the mice were removed and the connective tissue attached to the surface was removed as much as possible. Both ends of the long bone were pierced with a 2 ml disposable syringe and needle, the medullary cavity was washed with complete medium (DMEM / F12 containing 10% FBS), bone marrow cells were collected and blown to prepare a single cell suspension. Spread the cell suspension on the bottom of the plastic culture dish, and move it to the cell culture incubator for static culture. After 24 hours, the culture medium and the unattached cells contained therein were discarded (without washing), and an appropriate amount of medium was added to continue culturing. Thereafter, the medium was changed every 2 days an...

Embodiment 1

[0078] It is used to illustrate the influence of the fusion polypeptide provided by the present invention on the expression of CXCR4 and CXCR7 of mouse BMSC.

[0079] (1) Western blot detection of CXCR7, CXCR4 protein expression in cells treated

[0080] Take the BMSC cells prepared in Preparation Example 2 and inoculate them in 6cm dishes. The experiment is divided into control group (no polypeptide, only medium), and the experimental group is stimulated by adding different concentrations of polypeptide D (25, 50, 100, 150 μmol / L) for 12 hours. , cells were collected for Western blot detection. Each treatment was repeated 3 times.

[0081] Western blot detection: add protein lysate to the collected cells, let stand on ice for 10 minutes, then scrape off the cells, absorb the cell suspension and let stand for 10 minutes, centrifuge at 12,000 r / min for 10 minutes at 4°C. The supernatant was taken, and the total cell protein was separated by polyacrylamide gel electrophoresis ...

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Abstract

The invention relates to the field of medicine, and in particular relates to a fusion polypeptide, an application of the fusion polypeptide in enhancement of stem cell homing ability and anti-apoptotic ability and a pharmaceutical composition. The fusion polypeptide includes an SDF-1 derived peptide, an optional linker sequence and an RGD peptide having an RGD sequence in sequence from the 5' endto the 3' end, wherein the linker sequence is used to link the SDF-1 derived peptide and the RGD peptide, the SDF-1 derived peptide can specifically bind to CXCR4 and / or CXCR7 receptors expressed by stem cells, and the RGD peptide can specifically bind to an integrin receptor. The fusion polypeptide has the advantages of low costs and a long half-life period, and can enhance the homing ability andanti-apoptotic ability of the stem cells.

Description

technical field [0001] The invention relates to the field of medicine, in particular to a fusion polypeptide and its application and pharmaceutical composition in enhancing stem cell homing ability and anti-apoptosis ability. Background technique [0002] A large number of clinical practice and animal experiments have used stem cells for tissue defect and wound repair, aging-related diseases, ischemic diseases, immune-mediated diseases (such as autoimmune diseases, sepsis) and rejection after organ transplantation and other diseases, have shown good therapeutic effect. So far, although the clinical application of stem cell therapy is still in the exploratory stage, there is no doubt that compared with traditional treatments, stem cell therapy has shown more advantages. Therefore, stem cell transplantation therapy has shown great clinical application prospect and value, making it a hopeful strategy for the treatment of many diseases at present. Stem cells are divided into e...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61K38/18A61P17/02A61P39/06A61P9/10A61P37/00A61P37/06
CPCC07K14/475C07K5/0817A61P17/02A61P39/06A61P9/10A61P37/00A61P37/06C07K2319/00A61K38/00
Inventor 黄宏
Owner 中国人民解放军陆军特色医学中心
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