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Mptx2 protein and application thereof in treating methicillin-resistant staphylococcus aureus infection related diseases

A protein and application technology, applied in the fields of peptide/protein components, medical preparations containing active ingredients, antibacterial drugs, etc., can solve the problems of poor treatment effect, high antibiotic residues, imperfection, etc.

Pending Publication Date: 2022-04-19
SHANGHAI INST OF PEDIATRIC RES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, antibiotic treatment is the main means of treating MRSA infection, but because MRSA is multi-drug resistant to many antibiotics, the treatment effect is poor, and it is one of the most difficult clinical problems.
Moreover, due to the immature development of the immune system and weak immune function of children, the antibiotic residues in the body are high, and there are many kinds of antibiotic residues, which is extremely unfavorable for the growth and development of children.

Method used

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  • Mptx2 protein and application thereof in treating methicillin-resistant staphylococcus aureus infection related diseases
  • Mptx2 protein and application thereof in treating methicillin-resistant staphylococcus aureus infection related diseases
  • Mptx2 protein and application thereof in treating methicillin-resistant staphylococcus aureus infection related diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Example 1. Mptx2 protein expression and purification

[0084] 1.1 Main reagents

[0085] (1) Tryptone: ST800, Shanghai Biyuntian Biotechnology Co., Ltd.

[0086] (2) Yeast paste: ST968, Shanghai Biyuntian Biotechnology Co., Ltd.

[0087] (3) Sodium chloride: 1249KG001,

[0088] (4) High-purity low-electroosmosis agarose: G5056-100G, Wuhan Sewell Biotechnology Co., Ltd.

[0089] (5) pET-28 carrier: Shanghai Jikai Gene Chemical Technology Co., Ltd.

[0090] (6) BL21(DE3) chemically competent cells:: C504-03, Nanjing Novozyme Biotechnology Co., Ltd.

[0091] (7) Kanamycin: E004000-5G, Shanghai Yuanxiang Medical Instrument Co., Ltd.

[0092] (8) Petri dish 100×20mm TC surface PS (polystyrene) material: 430167, Corning Display Technology Co., Ltd.

[0093] (9) Isopropyl-β-D-thiogalactoside (IPTG) / Isopropylβ-D-thiogalactoside: MB3026, Shanghai Yuanzhi Biotechnology Co., Ltd.

[0094] (10) Ni-NTA Sefinose(TM) Resin Kit: C600332-0001, Sangon Bioengineering (Shanghai) Co....

Embodiment 2

[0163] Example 2. Mptx2 protein inhibits MRSA growth experiment in vitro

[0164] 2.1 Main materials

[0165] (1) Staphylococcus aureus: ATCC-6538, Beijing Biobowell Biotechnology Co., Ltd.

[0166] (2) MRSA: ATCC-43300, Beijing Biobowell Biotechnology Co., Ltd.

[0167] 2.2 Method

[0168] 2.2.1 Recovery culture of MRSA bacteria

[0169] (1) Add 100 μl of MRSA bacteria liquid to 1 L of liquid LB medium containing 50 μg / ml kanamycin. Place on a shaker for overnight culture (37°C, 220rpm, 16h).

[0170] (2) After culturing to the logarithmic growth phase, collect the bacterial pellet by centrifugation (4°C, 4000rpm, 15min).

[0171] (3) Resuspend the bacterial pellet with sterilized PBS buffer solution, and determine the total number of bacterial colonies in the bacterial liquid by plate counting method. Store at -80°C for later use.

[0172] (4) Before use, dilute MRSA with sterile PBS buffer to a final concentration of 5×10 5 CFU / ml.

[0173] 2.2.2 Mptx2 protein conc...

Embodiment 3

[0186] Example 3. Intervention of Mptx2 protein in MRSA infection-associated sepsis in mice

[0187] 3.1 Main materials

[0188] (1) C57BL / 6 mice: Shanghai Jihui Experimental Animal Breeding Co., Ltd.

[0189] (2) 1ml syringe: YA0550, Shanghai Jingkuang Biotechnology Co., Ltd.

[0190] 3.2 Method

[0191] 3.2.1 Mptx2 protein concentration adjustment

[0192] The drug concentration was adjusted to 1 mg / ml with sterile PBS buffer. All proteins were sterilized by filtration with a 0.22 μm filter membrane and stored in sterile 1.5ml centrifuge tubes before use.

[0193] 3.2.2 Recovery of MRSA bacteria solution

[0194] Add 20 μl of MRSA bacteria solution to 20ml of liquid LB medium, and place all the liquid in a sterile 50ml centrifuge tube. Place on a shaker and shake for 16 hours to the logarithmic growth phase (37°C, 220rpm). The bacterial liquid was collected by centrifugation (3000rpm, 15min, normal temperature). Use 1ml PBS to resuspend the bacteria solution, and det...

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Abstract

The invention provides an Mptx2 protein and application of the Mptx2 protein in treating diseases related to methicillin-resistant staphylococcus aureus infection.

Description

technical field [0001] The present invention relates to the field of new drug applications, in particular to the Mptx2 protein and its application in treating diseases related to methicillin-resistant Staphylococcus aureus infection. Background technique [0002] Staphylococcus aureus (Staphylococcus aureus) is an important human pathogen and the main cause of infectious diseases in humans and animals. Staphylococcus aureus can secrete a variety of virulence factors to cause infection in various tissues of the human body. With the widespread use of antibiotics, drug-resistant genes appeared in Staphylococcus aureus, resulting in methicillin-resistant Staphylococcus aureus (MRSA). [0003] MRSA produces varying degrees of drug resistance to a variety of antibiotics by changing the targets of antibiotics and producing modified enzymes. The detection rate of MRSA is increasing year by year, and the proportion of MRSA to Staphylococcus aureus in many areas even exceeds 80%, wh...

Claims

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Application Information

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IPC IPC(8): C07K14/47A61K38/17A61P31/04A61P1/00A61P1/16A61P13/12
CPCC07K14/47A61K38/1709A61P31/04A61P1/16A61P1/00A61P13/12Y02A50/30
Inventor 肖永陶蔡威王伟鹏朱景
Owner SHANGHAI INST OF PEDIATRIC RES
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