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Hybrid peptide with immunomodulatory and anti-inflammatory functions and preparation method and application thereof

An immunomodulatory, hybrid peptide technology, applied in the field of genetic engineering and biological preparations, can solve the problems of increasing the complexity and antagonism of clinical drug selection, failing to improve the body's immune function, and reducing immunity and production performance, and achieving weight reduction. As well as intestinal damage, good immune and anti-inflammatory two-way regulation, and the effect of relieving tissue damage

Active Publication Date: 2019-08-16
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are obvious problems in the use of antibiotics or hormonal anti-inflammatory drugs in the treatment of infection and inflammation: although antibiotics can reduce or kill pathogenic bacteria, they cannot improve the immune function of the body. On the contrary, the bacteria killed by antibiotics may also produce endotoxin or Exotoxins, which in turn aggravate the inflammatory response and even lead to systemic inflammatory response syndrome (Botwinski, 2001), which can cause fever, anorexia, excessive energy consumption in the body, decomposition of body tissues, and decline in immunity and production performance in animals. Avian mortality (Botwinski, 2001; Zinner, 1999)
Therefore, in the field of traditional medicine, the clear separation of anti-inflammatory drugs and immune-enhancing drugs does not meet the actual needs of the body or the interaction between anti-inflammatory and immune, and greatly increases the complexity and antagonism of clinical drug selection.

Method used

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  • Hybrid peptide with immunomodulatory and anti-inflammatory functions and preparation method and application thereof
  • Hybrid peptide with immunomodulatory and anti-inflammatory functions and preparation method and application thereof
  • Hybrid peptide with immunomodulatory and anti-inflammatory functions and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Preparation of immune anti-inflammatory hybrid peptide CTP

[0059] 1. Obtaining the hybrid peptide sequence

[0060] By studying the sequence, structure and relationship between sequence structure and function of polypeptide CATH-2 (sequence shown in SEQ ID NO.2) and thymosin TP5 (sequence shown in SEQ ID NO.3), protein molecular design technology was used to The hybridization of polypeptide CATH-2 and thymosin TP5 is carried out to obtain multiple candidate hybrid peptides. In this example, candidate hybrid peptides CTP (sequence shown in SEQ ID NO.1), TP5-CATH2 (sequence shown in SEQ ID NO.4) and CATH2-TP5' (sequence shown in SEQ ID NO.5) were used As an example, the screening process of immune anti-inflammatory hybrid peptide CTP is illustrated.

[0061] Take advantage of the properties of myeloid differentiation protein-2 (MD-2) (i.e., MD-2 binds to TLR4, confers TLR4 responsiveness to various ligands including LPS; and MD-2 can promote the expression o...

Embodiment 2

[0075] Example 2 Effect of Hybrid Peptide CTP on Survival Rate of Mouse Macrophage Cells

[0076] The macrophage RAW264.7 in the logarithmic growth phase was inoculated in a 96-well plate, and the initial cell culture density was 1×10 4 cells / mL, 100 μL per well, at 37°C, 5% CO 2 After culturing overnight under the same conditions, a series of concentration gradients of CTP, CATH2 and TP5 (0-100 μg / mL) were added, and after culturing for 24 hours, the effects of hybrid peptide CTP and its maternal peptide CATH2 and TP5 on mouse giant Effect on phagocyte survival. The result is as Figure 5 As shown, the cytotoxicity of polypeptide CTP is significantly lower than that of its parent peptide CATH2, and polypeptide CTP has no cytotoxicity at a concentration of 10 μg / mL, which is beneficial for subsequent research and application.

Embodiment 3

[0077] Example 3 Immunomodulatory activity of hybrid peptide CTP in mouse macrophages

[0078] The hybrid peptide CTP and its parental peptides CATH2 and TP5 were diluted with DMEM medium to prepare a peptide solution with a concentration of 10 μg / mL, and the effects of CTP and its parental peptides CATH2 and TP5 on normal and LPS-induced inflammatory conditions were detected. Effects on the secretion of TNF-α, IFN-γ and other cytokines in mouse macrophage RAW264.7. Set normal group (Control), model group (LPS), test group 1 (CATH2), test group 2 (TP5), test group 3 (CTP) and test group 4 (CATH2+LPS), test group 5 (TP5+LPS) ) test group 6 (CTP+LPS), wherein, the normal group did not do any treatment; test groups 1, 2, and 3 were respectively added with a final concentration of 10 μg / mL CATH2, TP5 or CTP solution after the cells were cultured overnight; test groups 4, 5 and 6 respectively added CATH2, TP5 or CTP solutions with a final concentration of 10 μg / mL after the cells ...

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Abstract

The invention relates to the technical field of genetic engineering and biological preparation, in particular to a hybrid peptide having immunomodulating and anti-inflammatory functions, and a preparation method and an application thereof. The invention provides an immuno-anti-inflammatory hybrid peptide CTP, which is obtained by hybridization, optimization and screening of antibacterial peptide CATH-2 and thymosin TP5, and an amino acid sequence is shown as SEQ ID NO. 1. The hybrid peptide CTP has high bidirectional immunomodulatory activity, can enhance the immune function of the body undernormal or immunosuppressed state, and protects the body from damage caused by immunosuppression; in the inflammatory state, CTP can also inhibit the body's inflammatory response, alleviates the damageof an inflammatory reaction on the tissue, and has the advantages of low cytotoxicity, high safety, simple preparation method and low cost, can be used as an ideal immuno-anti-inflammatory regulator,and has good application potential and value.

Description

technical field [0001] The invention relates to the fields of genetic engineering and biological preparations, in particular to a hybrid peptide with immune regulation and anti-inflammatory functions, its preparation method and application. Background technique [0002] When animals or humans are young, weak, disease, stress, pathogenic infection, etc., their immunity is often reduced, and then secondary infection (bacterial or viral mixed infection) occurs, causing inflammatory reactions (red, hot, swollen) , pain, etc.). At present, for the treatment of infection and inflammation, the traditional and commonly used prevention and treatment strategy is to use antibiotics and hormone anti-inflammatory drugs (such as hydrocortisone, dexamethasone, etc.), although it can effectively control infectious inflammation and non-infectious inflammation, However, continuous use can cause various side effects: such as serious disturbance of water and salt metabolism, sugar, fat, and pr...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/81A61K38/17A61K38/32A61P37/04A61P29/00A23L33/18A23K20/147
CPCA23V2002/00A61K38/00A23K20/147A23L33/18A61P29/00A61P37/04C07K14/4723C07K14/57581C12N15/815C12N2800/22A23V2250/55A23V2200/30A23V2200/324
Inventor 张日俊张璐璐卫旭彪黄燕阿曼斯大勇李仲玄程俊豪杜孟思
Owner CHINA AGRI UNIV
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