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Novel septicemia polypeptide and application thereof to diagnosis of septicemia

A technology of peptides and uses, which is applied in the field of new sepsis diagnostic indicators, can solve the problems that cannot meet the needs of acute and severe sepsis diagnosis and treatment, it is difficult to distinguish infectious from non-infectious, and the time required for bacterial culture is long.

Active Publication Date: 2017-01-11
XIAMEN LUJIA BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current gold standard for laboratory diagnosis of sepsis is bacterial culture, but bacterial culture takes a long time and cannot meet the needs of diagnosis and treatment of severe sepsis
C-reactive protein (CRP) and IL-6 are also used in the auxiliary diagnosis of sepsis, but both of them are also significantly increased in SIRS, and it is difficult to distinguish between infectious and non-infectious

Method used

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  • Novel septicemia polypeptide and application thereof to diagnosis of septicemia
  • Novel septicemia polypeptide and application thereof to diagnosis of septicemia
  • Novel septicemia polypeptide and application thereof to diagnosis of septicemia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0165] Example 1 Synthesis, separation, purification and identification of DcR3 active polypeptide and its P1 derivative polypeptide

[0166] The DcR3 active polypeptide and its P1 derivative polypeptide were synthesized by a commercial peptide synthesis company according to conventional methods, separated and purified by HPLC routinely, and the molecular weight and purity were identified by mass spectrometry and HPLC. Purity>98% is qualified product, freeze-dried and stored for future use.

[0167] DcR3 active polypeptide can form a disulfide bond from Cys in the liquid to form a dimer, and the dimer is purified after identification.

Embodiment 2

[0168] Example 2 DcR3-P1 polypeptide affinity experiment

[0169] The SPR biosensing technology of BiaCore system was used to identify the affinity of the purified DcR3 active polypeptide and DcR3 active polypeptide dimer to FasL ligand. Use the amino cross-linking kit to covalently cross-link FasL to the gold surface on the CM5 reaction chip, and then let the DcR3 dimer flow through the gold surface at a speed of 5ul / min for 10 minutes to wash away unbound substances, and pass Determination of plasmon resonance signal (response units, RU) and bioinformatics analysis (BIA) evaluation calculated Kd (dissociation constant, nM). result( Figure 8 ) shows that binding to the ligand is visible at a concentration of 20 ng / ml. The Kd value of DcR3 binding to FasL is 271.4 + 24.4nM 28 . It can be speculated that DcR3-P1 can compete with the pro-apoptotic Fas receptor for the biological activity of its ligand FasL, thereby blocking the apoptosis-inducing function of FasL. FasL is p...

Embodiment 3

[0170] Example 3 Inhibitory effect of DcR3-P1 polypeptide on sepsis

[0171] After confirming that DcR3-P1 has a certain anti-apoptotic function at the cellular level, this experiment carried out a preliminary experiment on the antagonism of LPS sepsis in mice.

[0172] Considering the absorption and action time, DcR3-P 10mg / ml (control group) or 10mg / kg / 0.2ml saline (experimental group) i.v. injected into BALB / c mice (6 / group, a small amount of initial test), 10 minutes later , LPS (2mg / kg, E.coli 0111:B4, Sigma) was intraperitoneally injected into the mice of each group, and then the development and changes of sepsis were observed.

[0173] The mice in the control group showed hair erection all over the body within a few hours, lethargy, lack of movement, no food, dull eyes, blocked microvessels and congestion, similar to DIC (disseminated intravascular coagulation) symptoms: obvious symptoms on the nose, face, ears, limbs, and tail Cyanosis, swollen head and face, compared...

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PUM

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Abstract

The invention provides a DcR3 polypeptide for treating infectious diseases and applications thereof. Concretely, DcR3 is a marker for indicating infectious inflammation with high specificity and sensitivity, and inhibition of DcR3 has effective treatment effects for infectious inflammation. The polypeptide of DcR3 is designed and synthesized for competitive inhibition of combination of apoptosis acceptors (such as Fas, HVEM / LT R, DR3, etc.) and apoptosis factors (FasL, LIGHT, TL1A, etc.), so that apoptosis pathway is blocked, and biological effects of infectious inflammation (such as septicemia) symptoms can be effectively reduced. In addition, experiments also show that when DcR3 and PCT are used as markers at the same time, specificity and sensitivity of early diagnosis of septicemia are improved.

Description

technical field [0001] The present invention belongs to the technical field of medical diagnosis and treatment. Specifically, the present invention relates to the application of polypeptide derived from DcR3 and its derivatives in the treatment of sepsis and the blood DcR3 required for the treatment as a new diagnostic index of sepsis. Background technique [0002] Sepsis is a serious infectious disease in which pathogenic bacteria enter the blood. The pathogenic bacteria that enter the blood multiply and grow at an astonishing rate of one generation per 20 minutes, spread rapidly, and can develop into septic shock, disseminated intravascular coagulation (DIC) and multiple organ failure within a few hours, with a mortality rate of 25-46 %, so it is a clinical emergency. Early correct diagnosis and effective treatment are the key to rescuing patients. [0003] The pathophysiological mechanism of sepsis is that pathogenic bacteria and their toxins enter the blood and activat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/715C12N15/12A61K38/17A61P29/00A61P31/00G01N33/68
CPCA61K38/00C07K14/7151G01N33/6812G01N2800/26
Inventor 张鲁榕张美张世民张震寰林建华欧启水杨滨高丽钦张海荣金顺姬
Owner XIAMEN LUJIA BIOTECH
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