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Application of human sDR5-Fc (soluble DR5-Fc) antibody fusion protein used as renal injury treatment medicament

A technology of fusion protein and therapeutic drug, which is applied in the field of medicine to achieve the effect of protecting the kidney and reducing the degree of damage

Inactive Publication Date: 2016-06-22
HENAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, whether sDR5 has a therapeutic effect on renal tissue damage caused by ischemia has not been reported at home and abroad.

Method used

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  • Application of human sDR5-Fc (soluble DR5-Fc) antibody fusion protein used as renal injury treatment medicament
  • Application of human sDR5-Fc (soluble DR5-Fc) antibody fusion protein used as renal injury treatment medicament
  • Application of human sDR5-Fc (soluble DR5-Fc) antibody fusion protein used as renal injury treatment medicament

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Example 1: Preparation of human sDR5-Fc antibody fusion protein

[0034] The DR5 extracellular segment gene sequence (182 amino acids, http: / / www.uniprot.org / uniprot / O14763) was cloned into the eukaryotic expression vector pGS-Fc with the human IgG Fc segment, and the designed primers were: sense (5' -3'): ggaagcttgccaccATGGAACAACGGGGACAG, antisense (5'-3'): aagaattcTCTGGTCGTGGTGCAGGG. Extract the high-purity plasmid (OMEGAPlasmidMidiKit) and adjust the concentration to 500ng / μl. Resuscitate CHO / K1 cells, adjust the cell state to the optimal state, and conduct transient electroporation (300v, interval time 0.125s, duration 0.1ms, electric shock three times), and obtain high-expression stable cell lines by MSX pressurized screening. At present, the yield of the fusion protein of the present invention can reach 1.9 g / L. The purified protein was used for functional verification in vitro to detect whether it could block TRAIL-induced apoptosis of the human leukemia cell l...

Embodiment 2

[0037] Example 2: Human sDR5-Fc antibody fusion protein protects against renal ischemia-reperfusion injury

[0038] Establishment of mouse animal model of renal ischemia-reperfusion injury

[0039] Clean grade male C57BL / 6 mice aged 6-8 weeks, after weighing, intraperitoneally inject 5% chloral hydrate (7.5ml / kg) for anesthesia, place the mice on a constant temperature table, remove the hair on the back of the mice, and expose the skin, After the body temperature of the mouse returned to 36°C, the operation began. Small openings of about 1 cm were opened below the ribs of the mouse and on both sides of the spine. The left kidney of the mouse was lower than the right kidney, so the small opening of the left kidney was slightly downward. Cut off the muscle layer, and gently squeeze the kidney out of the wound with a cotton swab to expose the two kidneys; the renal pedicle of the right kidney is slightly shorter, first clamp the right kidney with an arterial clamp, and then clamp...

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Abstract

The invention belongs to the field of medicine, and in particular relates to medicinal application of an sDR5-Fc (soluble DR5 and Fc) antibody fusion protein formed by fusing human sDR5 and an Fc segment of a human immunoglobulin, in particular to application of the human sDR5-Fc antibody fusion protein used as a renal injury treatment medicament. According to the invention, experiments prove that in a rat renal ischemia model and an ischemia-reperfusion model, the human sDR5-Fc antibody fusion protein can obviously reduce renal tissue injury, and obviously reduce an apoptosis rate of renal tubular epithelial cells. As a novel candidate medicament for treating acute kidney injury, the human sDR5-Fc antibody fusion protein has great developing potential.

Description

technical field [0001] The invention belongs to the field of medicine, and in particular relates to the medicinal application of human sDR5 (solubleeathreceptor5, sDR5) protein, especially the application of human sDR5-Fc antibody fusion protein as a first-aid and therapeutic drug for renal ischemia-reperfusion injury. Background technique [0002] Ischemia-reperfusion injury refers to the interruption or reduction of blood supply to an organ or tissue, resulting in local tissue or organ ischemia and hypoxia. And the phenomenon of further aggravation of metabolic disorders and institutional destruction. Clinically, ischemia-reperfusion injury is common in organ transplantation, partial organ removal, trauma, and shock. Due to the high perfusion characteristics of the kidney, the incidence of ischemia-reperfusion injury is relatively high. [0003] Transient ischemia can cause reversible damage to the kidney, while long-term ischemia-reperfusion can cause irreversible damage...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61P13/12A61P9/00
CPCA61K38/177C07K2319/30
Inventor 马远方王志增张军王耀辉李小全
Owner HENAN UNIVERSITY
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