CN103479659A - Application of three kinds of oroxin in preparing drug for resisting staphylococcus aureus infection - Google Patents
Application of three kinds of oroxin in preparing drug for resisting staphylococcus aureus infection Download PDFInfo
- Publication number
- CN103479659A CN103479659A CN201310482112.0A CN201310482112A CN103479659A CN 103479659 A CN103479659 A CN 103479659A CN 201310482112 A CN201310482112 A CN 201310482112A CN 103479659 A CN103479659 A CN 103479659A
- Authority
- CN
- China
- Prior art keywords
- oroxin
- kinds
- staphylococcus aureus
- application
- aureus infection
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Abstract
The invention relates to application of three kinds of oroxin in preparing a drug for resisting staphylococcus aureus infection. The staphylococcus aureus infection resistance of three kinds of oroxin can be proved by virtue of a sheep red blood cell haemolysis test, molecular dynamics simulation and a human pulmonary epithelial cell (A549) injury protective test. Compared with treatment by antibiotics, treatment by using three kinds of oroxin has the characteristics of no drug resistance and high cure rate, so that the three kinds of oroxin can be used in development of new drugs, and has an important meaning to drug target confirmation.
Description
Technical field
The present invention relates to the application of three kinds of oroxins in the anti-golden Portugal of preparation bacterium infection medicine, belong to field of medicaments.
Background technology
These three kinds of oroxin (Oroxin A, Oroxin B, Oroxylin A 7-O-glucuronide) be isolated three kinds of natural flavone compounds from the dry mature seed of Bignoniaceae plant Semen Oroxyli, in the vegetables such as Fructus Fragariae Ananssae, Fructus Vitis viniferae, Fructus Mali pumilae, Bulbus Allii Cepae and fruit, existence also arranged.There is obvious antioxidation, anti-inflammatory property, immune system is also had to certain regulating action, but have no so far the report that fisetin infects for anti-golden Portugal bacterium both at home and abroad.
Staphylococcus aureus is one of clinical modal conditioned pathogen, surpasses in 50% adult human body and all carries golden Portugal bacterium, and wherein the crowd of 10-20% forever grows surely.Gold Portugal bacterium is except the local folliculitis that causes the people, furuncle and phyma, cellulitis, can also cause the life-threatening diseases such as the multiple suppurative illness such as pneumonia, pericarditis, osteomyelitis, pyelonephritis, renal abscess and bacteremia, endocarditis, there is very high M & M, as the case fatality rate by systemic infection due to golden Portugal bacterium up to more than 50%.In recent years, antibiotic a large amount of uses cause Bacterial Drug Resistance of Patients to increase, and have been found that the various clinical separated strain is serious to the tolerance of the antibiotic such as tetracycline and ciprofloxacin, cause the treatment of golden Portugal bacterium bacterium disease to face the situation pasted medical help.Therefore, find novel, safe medicine extremely urgent.These three kinds of oroxins come from China's Chinese medicine, and the present invention studies confirm that these three kinds of oroxins resist the bacterium infection of golden Portugal and have therapeutic effect preferably.
Summary of the invention
Molecular structure is as follows:
The present invention confirms its anti-golden Portugal bacterium bacterium infection effect by sheep red blood cell hemolytic test, huge protection test and the mice gold Portugal bacterium infection model of biting like cell J774 and Turnover of Mouse Peritoneal Macrophages damage of mice.
The accompanying drawing explanation
Fig. 1. the haemolysis titre of alpha hemolysin after not adding oroxin and adding the oroxin of variable concentrations.
Fig. 2. three kinds of oroxin combinations.
The specific embodiment
hemolytic test
The golden Portugal bacterium alpha hemolysin of purification is done continuous doubling dilution in 96 orifice plates, in each hole, adds 10
7individual sheep red blood cell, centrifugal hatch 30h in 37 ℃ after.Make haemoclastic highly diluted multiple be considered as haemolysis titre (Hemolysis titre), as shown in Figure 1.
the application molecular dynamics simulation is proved conclusively the anti-golden Portugal of oroxin bacterium alpha hemolysin avtive spot
2.1 application molecular dynamics simulation conclusive evidence avtive spot
(1) molecular docking (molecular docking)
For preliminary definite oroxin and alpha hemolysin binding site, and oroxin and the intermolecular binding ability power of alpha hemolysin, at first carry out the molecular docking of oroxin and alpha hemolysin, tentatively determine the structure of oroxin and alpha hemolysin complex.
(2) molecular dynamics simulation (Molecular dynamics simulations)
Use between molecular dynamics simulation research alpha hemolysin molecule and its mutant and oroxin in conjunction with can variation, in conjunction with conformational analysis methods analyst alpha hemolysin protein conformation, change, particularly in conjunction with the conformation change of the calmodulin binding domain CaM of oroxin substrate.Determine oroxin and alpha hemolysin protein binding avtive spot.
(3) fixed point residue sudden change (Site-directed mutagensis)
According to the result of molecular simulation, find pivotal role amino acid residue in oroxin and alpha hemolysin protein binding.The applied molecular biology method, to fixed a point residue sudden change of alpha hemolysin, clonal expression goes out the alpha hemolysin albumen of wild type and mutant respectively.
(4) binding constant of Fluorimetric Quenching Method for Determination protein and oroxin
By after oroxin and alpha hemolysin albumen (wild type and mutant) interaction, the binding constant of application Fluorimetric Quenching Method for Determination alpha hemolysin and oroxin, and compare with the result of dynamics simulation, determine the interactional action target spot of oroxin and alpha hemolysin.
2.2 mechanism analysis
According to calculating and Energy Decomposition in conjunction with free energy, analyze the conformation change of calmodulin binding domain CaM key amino acid residue, find out oroxin and alpha hemolysin protein binding ability specificity and the big or small factor of affecting, finally determine contacting between alpha hemolysin albumen and oroxin complex conformation change and drug effect.
2.3 experiment conclusive evidence
The mechanism of action of the anti-alpha hemolysin of alpha hemolysin oligomerization verification experimental verification oroxin that the application deoxycholic acid is induced.
Result shows, these three kinds of oroxins all can be incorporated into alpha hemolysin stem zone, suppress it from monomer to polymeric conformation change, thereby suppresses the hemolytic activity of alpha hemolysin, as shown in Figure 2.
three kinds of protection tests that oroxin damages the people's pulmonary epithelial cells (A549) by the mediation of golden Portugal bacterium alpha hemolysin
In golden Portugal bacterium and A549 co-culture of cells system, add the oroxin of variable concentrations to carry out common cultivation, then carry out following test and investigate oroxin pair
α-the protective effect of the A549 damage of hemolysin mediation.
Work/dead cell test (Live/Dead assay)
After adding live (green)/dead (red) reagent effect in above-mentioned coculture, observe the state of A549 cell under laser confocal microscope, it is green that living cells is, and dead cell takes on a red color.
Cell toxicity test (LDH assay)
The LDH activity of the above-mentioned coculture supernatant of application LDH kit measurement, the survival rate of calculating A549 cell, the protective effect of quantitative expedition oroxin to the A549 cell.
Result shows, people's pulmonary epithelial cells damage that these three kinds of medicines can protect golden Portugal bacterium hemolysin to mediate, and this effect presents dose dependent.
Claims (4)
1. three kinds of oroxin application in the anti-golden Portugal of preparation bacterium infection medicine.
2. medicine comprises pharmaceutically acceptable dosage form as claimed in claim 1.
3. the infection of golden Portugal bacterium refers to by the bacterial bacterial infection of golden Portugal Pseudomonas as claimed in claim 1.
4. bacterial infection refers to the infection caused by staphylococcus aureus as claimed in claim 3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310482112.0A CN103479659A (en) | 2013-10-16 | 2013-10-16 | Application of three kinds of oroxin in preparing drug for resisting staphylococcus aureus infection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310482112.0A CN103479659A (en) | 2013-10-16 | 2013-10-16 | Application of three kinds of oroxin in preparing drug for resisting staphylococcus aureus infection |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103479659A true CN103479659A (en) | 2014-01-01 |
Family
ID=49820480
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310482112.0A Pending CN103479659A (en) | 2013-10-16 | 2013-10-16 | Application of three kinds of oroxin in preparing drug for resisting staphylococcus aureus infection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103479659A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105232568A (en) * | 2014-07-10 | 2016-01-13 | 苏州大学 | Application of oroxin B and drug containing oroxin B |
| CN110139647A (en) * | 2016-11-11 | 2019-08-16 | 萨米莱布斯有限公司 | Composition and its extracting method comprising oroxylin A |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102727509A (en) * | 2011-04-13 | 2012-10-17 | 吉林大学 | Application of baicalin to preparation of medicament for treating pneumonia |
-
2013
- 2013-10-16 CN CN201310482112.0A patent/CN103479659A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102727509A (en) * | 2011-04-13 | 2012-10-17 | 吉林大学 | Application of baicalin to preparation of medicament for treating pneumonia |
Non-Patent Citations (3)
| Title |
|---|
| 殷文光 等: "木蝴蝶的研究进展", 《中国中药杂志》, vol. 32, no. 19, 31 October 2007 (2007-10-31), pages 1965 - 1970 * |
| 王锐 等: "木蝴蝶的化学成分和药理作用", 《广东农业科学》, no. 22, 31 December 2011 (2011-12-31), pages 121 - 123 * |
| 胡庭俊 等: "木蝴蝶提取物制备及其抗菌抗炎活性的研究", 《中国畜牧兽医》, vol. 37, no. 3, 31 December 2010 (2010-12-31), pages 225 - 228 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105232568A (en) * | 2014-07-10 | 2016-01-13 | 苏州大学 | Application of oroxin B and drug containing oroxin B |
| CN105232568B (en) * | 2014-07-10 | 2018-03-27 | 苏州大学 | The application of oroxin B and the medicine containing oroxin B |
| CN110139647A (en) * | 2016-11-11 | 2019-08-16 | 萨米莱布斯有限公司 | Composition and its extracting method comprising oroxylin A |
| CN110139647B (en) * | 2016-11-11 | 2022-11-01 | 萨米莱布斯有限公司 | Composition containing oroxylin A and extraction method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Agarwal et al. | Comparative analysis of antibacterial activity of four Piper betle varieties | |
| Channa et al. | Effect of oral supplementation of Aloe vera extract on haematology indices and immune cells of blood in rabbit | |
| CN103479659A (en) | Application of three kinds of oroxin in preparing drug for resisting staphylococcus aureus infection | |
| CN103690538B (en) | With the active Chinese medical extract of anti-poultry mycoplasma, Chinese medicine composition | |
| Ou et al. | Dryocrassin ABBA, a novel active substance for use against amantadine-resistant H5N1 avian influenza virus | |
| Ali | In vitro studies of antimicrobial activity of (Curcuma longa L.) rhizomes against helicobacter pylori | |
| CN103271852B (en) | Massage oil containing prinsepia utilis royle oil and preparation method of massage oil | |
| CN103169652B (en) | Veterinary alkaline lincomycin injection as well as preparation method and use thereof | |
| CN103989693B (en) | Application of scutelloside in preparation of acute hemolytic uremic syndrome treatment drugs | |
| CN104586862A (en) | Application of N-acetylneuraminic acid monomer, N-acetylneuraminic acid hydrate or N-acetylneuraminate in external anti-inflammation medicines | |
| CN103585277A (en) | Application of clove leaf extractives in inhibition of streptococcus suis and intervention and elimination of streptococcus suis biofilm | |
| CN104147044B (en) | Composition for treating piglet staphylococcal exudative epidermitis | |
| CN102872004A (en) | Application of naringenin in preparation of medicine for curing pneumonia | |
| CN102872005B (en) | Application of the glycyrrhizin in treatment pneumonia medicine is prepared | |
| CN102379958A (en) | Chinese medicinal preparation for resisting bacteria, diminishing inflammation, eliminating phlegm and relieving cough and preparation method thereof | |
| CN105998029A (en) | Antibacterial composite drug of levofloxacin hydrochloride for injection | |
| CN102600159A (en) | Composition comprising tetrandrine, tetrandrine-based ramification and fluoroquinolones and application of composition | |
| CN108670951A (en) | A kind of compound sustained-released injection of gentamicin sulphate-Lincomycin Hydrochloride for animals and preparation method thereof | |
| Xu et al. | Cortex cercis chinensis granules attenuate Streptococcus pneumoniae virulence by targeting pneumolysin | |
| CN102429899A (en) | Application of baicalein to preparation of medicament for treating pneumonia | |
| Chaudhary | Antiacne activity of some Indian herbal drugs | |
| CN102727509A (en) | Application of baicalin to preparation of medicament for treating pneumonia | |
| YAN et al. | Influence of total glucosides paeony on the expression of IFN-γand IL-10 in peripheral blood of patients with oral lichen planus | |
| CN103191099B (en) | The application in anti-listeria infection medicine prepared by fisetin | |
| CN102872003A (en) | Application of apigenin on preparation of medicine for treating pneumonia |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140101 |