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Ischemic-myocardium-targeted VEGF (vascular endothelial growth factor) recombinant protein

A protein and protein technology, applied in the medical field, can solve the problems of side effects and dose dependence of VEGF

Pending Publication Date: 2017-01-04
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in clinical trials, VEGF recombinant protein showed a dose-dependent effect due to its easy diffusion (Hendel et al., 2000; Henry et al., 2003; Jacquier et al., 2007; Lopez et al., 1998), while High concentrations of VEGF may have side effects (Lee et al., 2000; Matsuno et al., 2002)

Method used

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  • Ischemic-myocardium-targeted VEGF (vascular endothelial growth factor) recombinant protein
  • Ischemic-myocardium-targeted VEGF (vascular endothelial growth factor) recombinant protein
  • Ischemic-myocardium-targeted VEGF (vascular endothelial growth factor) recombinant protein

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Embodiment 1, the acquisition of fusion protein IMT-VEGF

[0058] 1. Fusion protein IMT-VEGF

[0059] Fusion protein IMT-VEGF including VEGF 165 and the targeting polypeptide IMT and the connecting peptide connecting the two;

[0060] The fusion protein IMT-VEGF can also include VEGF 165, connecting peptide, targeting polypeptide IMT and 6 His tags ( figure 1 A).

[0061] The amino acid sequence of the fusion protein IMT-VEGF is sequence 2, and the 1-166th position of sequence 2 is VEGF 165 1. The 167-177th position of sequence 2 is the connecting peptide, the 178-186th position of the sequence 2 is the targeting polypeptide IMT, the 187-188th position of the sequence 2 is the amino acid residue encoded by the XhoI restriction site, the 189th-189th position of the sequence 2 194 bits are 6 His tags.

[0062] The nucleotide sequence of the fusion protein IMT-VEGF coding gene is sequence 1, and the 4th-498th position of sequence 1 is VEGF 165 Coding gene, the 499-5...

Embodiment 2

[0081] Embodiment 2, the biological activity detection of fusion protein IMT-VEGF

[0082] 1. IMT-VEGF stimulates endothelial cell proliferation

[0083] Primary cultured human umbilical vein endothelial cells (Merck Millipore, SCCE001) were inoculated into a 48-well plate, and the purified IMT-VEGF obtained in Example 1 and VEGF as a control were diluted to different gradient concentrations and added to the 48-well plate respectively. . MTT assay was performed after 72 hours of culture.

[0084] The result is as figure 1 As shown in C, there is no significant difference in the ability of IMT-VEGF and VEGF to stimulate the proliferation of endothelial cells, indicating that IMT-VEGF fused with the targeting polypeptide does not affect the biological activity of VEGF itself.

[0085] 2. Detection of targeting ability of IMT-VEGF

[0086] 1) The content of VEGF in normal myocardium, ischemic myocardium and peripheral blood serum

[0087] Construct the rat heart ischemia-rep...

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Abstract

The invention discloses an ischemic-myocardium-targeted VEGF (vascular endothelial growth factor) recombinant protein. The invention provides a fusion protein which is composed of a VEGF 165, a targeted polypeptide IMT and a connecting peptide for connecting the VEGF 165 and targeted polypeptide IMT. The experiment proves that the brand-new ischemic-myocardium-targeted VEGF protein IMT-VEGF designed in the invention can be targeted to the infarct zone through intravenous injection so as to increase the VEGF effective local concentration. The ischemic-myocardium-targeted VEGF recombinant protein has wide application prospects in targeted therapy, and can be targeted to the ischemic myocardium tissues through intravenous injection to promote the injury repair and cardiac function restoration.

Description

technical field [0001] The invention belongs to the medical field, and in particular relates to a construction method and application of a VEGF recombinant protein targeting ischemic myocardium. Background technique [0002] Myocardial infarction (MI) is one of the diseases with the highest morbidity and mortality in the world (Go et al., 2014), which seriously threatens human health and survival, and there is no effective method for the treatment of myocardial infarction . Reconstructing the blood circulation system at the infarction site and inducing myocardial regeneration are very important aspects in the treatment strategy of myocardial infarction. Clinically, angioplasty or collateral surgery is mainly used to restore blood supply (van der Spoel et al., 2011), and therapeutic angiogenesis induced by growth factors is also a very promising method. Vascular endothelial growth factor (vascular endothelia growth factor, VEGF) is one of the most effective cytokines to pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62A61K38/18A61K47/48A61P9/00A61P9/10
Inventor 戴建武杨赟陈冰侯祥林肖志峰赵燕南
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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