CN101164550A - Application of realgar microorganism leaching liquid in preparing antibiotic and antiviral medicine - Google Patents
Application of realgar microorganism leaching liquid in preparing antibiotic and antiviral medicine Download PDFInfo
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- CN101164550A CN101164550A CNA2007102020851A CN200710202085A CN101164550A CN 101164550 A CN101164550 A CN 101164550A CN A2007102020851 A CNA2007102020851 A CN A2007102020851A CN 200710202085 A CN200710202085 A CN 200710202085A CN 101164550 A CN101164550 A CN 101164550A
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Abstract
The present invention relates to an application of realgar microbial extract in preparation of medicine for resisting bacteria and resisting various. The realgar microbial extract is prepared according to the microbial treatment method disclosed by Chinese invention patent application 20061002000675. In the lowest arsenic concentration range (microgram grade) said extract has obvious effect for inhibiting colibacillus, staphylococcus aureus, candida albicans, bacillus subtilis, epidermal drug-resistant staphylococcus, comprodeum bacillus, non-drug-resistant epidermal staphylococcus, waxy epidermal staphylococcus, aeruginose pseudomonads and herpesvirus, etc. so that it can be used for preparing medicine for resisting bacteria and resisting virus., and can be made into various dosage forms, including suppository, pill preparation, granules preparation, film preparation, microcapsule preparation, dipping pills preparation, aerosol, tincture preparation, syrup, oral liquor and injection, etc.
Description
Technical field
The present invention relates to realgar microorganism extracting liquid in application antibiotic, anti-virus aspect.
Background technology
Main Red Arsenic Sulfide (the As that contains of Realgar (Realgar)
4S
4), there is the people to think that also it is realgar (As that its master contains thing
2S
2), have another name called gold stone, be common minerals as Chinese medicine.Since Shennong's Herbal, in the medical history in existing two over thousands of years of China.Realgar has effects such as detoxifcation parasite killing, removing dampness and eliminating phlegm, preventing the attack (or recurrence) of malaria, is mainly used in carbuncle furuncle, snakeworm mosquito wound, abdominal pain due to worm stagnation, infantile convulsion, malaria etc.
[1], modern medicine thinks that Realgar has effect antibacterial, that sterilization suppresses virus, has been successfully used to the treatment of various skin disease clinically
[2,3,4,5], therefore more and more be subjected to the extensive concern of Chinese scholars, become a focus of mineral Chinese medicine research.Mineral drug is the culture rarity of the Chinese nation, but because its water-insoluble, causes bioavailability low, problems such as clinical consumption is big, toxicity height, the development that has seriously restricted mineral Chinese medicine with crack the international market.Realgar is the important component of many Chinese patent medicines as a kind of important mineral Chinese medicine, and the export trade of China's Chinese patent medicine is obstructed repeatedly because of Hg and As equal size exceed standard.Therefore, find out a kind of effectively stripping mineral drug effective ingredient, reduce the mineral drug use amount, increase curative effect, to reduce toxic mineral drug concocting method significant.
Compare with traditional processing procedure, the preparation of Realgar ultramicro powder and nanometer grade powder and use have increased the dissolution characteristic of medicine on certain degree, thereby make the aspect such as absorption, distribution, bioavailability of Realgar all obtain raising in various degree, strengthened drug effect.But this class is handled and still can only be used as medicine with suspension, remains the GI irritation that causes oral, takes problems such as difficulty, and produce nanometer-size realgar needs main equipment and advanced technology in addition, and universal use is still had any problem
[6,7]
At present, a high-new biotechnology microbial metallurgy technology is a dark horse
[8-10]Under certain air, pH, temperature conditions, the Ore lysate that some special antibacterials (thiobacillus ferrooxidant, thiobacillus thiooxidans etc.) can be changed insoluble sulphide ore into solubility reclaims required element then
[11-13]The maximum characteristics of this technology are simple, efficient, low-cost, pollution-free, mild condition, thereby metallurgical industry is extensively praised highly and used in the world, and 30% of world's copper output from the microbial metallurgy technology at present
[14]Realgar also is a kind of sulphide ore, have by the dissolved possibility of leaching microbacteria, Chinese invention patent 2006102000675 discloses the microbial treatment method of mineral Chinese medicines such as Cinnabaris and Realgar, and the present invention relates generally to the Realgar leachate of microbial treatments in application antibiotic, anti-virus aspect.Experiment shows: this leachate can (Gamma Magnitude) have the obvious suppression effect to comprising escherichia coli, staphylococcus aureus, Candida albicans, bacillus subtilis, epidermis drug resistance staphylococcus, excrement enterobacteria, non-drug resistance staphylococcus epidermidis, wax shape staphylococcus epidermidis, pseudomonas aeruginosa, herpesvirus etc. in extremely low arsenic concentration scope.The reduction of arsenic and other harmful substance amount has obviously reduced the toxicity of Realgar, and drug effect increases simultaneously.
List of references
(1) Pharmacopoeia of the People's Republic of China version in 2005 is one one
(2) Kang Yong, Li Xianrong, Cheng Xia, Li Huijuan.The experimentation of Realgar pharmacological action and toxicity thereof are observed.The time precious traditional Chinese medical science traditional Chinese medicines, 1998,9 (4): 322-323.
(3) Yuan Chang comes, Dong Faqin.The research of the antibacterial anophelifuge elementary material of Chinese medicine.Environment and occupational medicine, 2006,23 (5): 399-405.
(4) Qin Yuqing.Health lotion for skin care EXPERIMENTAL OBSERVATION ON BACTERICIDAL EFFICACY.The medical faunae control, 2003,19 (11): 677-678.
(5) Liu Yibo, red legend, Zhan Yunli.The external bacteriostasis of Realgar lotion is investigated.The practical combination of Chinese and Western medicine is clinical, and 2002,2 (2): 49-50.
(6) Zhan Xiuqin.Modernization of Chinese medicine technology of preparing-professor Guo Liwei who grows with each passing hour talks nanometer Chinese medicine and micron Chinese medicine.Beijing University of Chinese Medicine's journal (natural science edition) .2002,18 (4): 198-201.
(7)Ying?D.，Huibi?X.et?al.Size?effects?of?realgar?particles?on?apoptosisin?a?human?vein?endothelial?cell?line：ECV-304.Pharmacological?Research，2001，44：513-519.
(8)Torma?A?E.The?role?of?Thiobacillus?ferrooxidans?in?hydrometallurgialprocesses.Adv?Biochem?Eng.1977，6：1-37
(9)Colmer?A?R，Hinckel.M?E.The?role?of?microorganism?in?acid?minedrainage.A?preliminary?report，Science，1947，106：253-256.
(10)Yuan，X，et?al.Microbes?processing?technology?study?for?non-metallicmineral?s.China?Non-Metallic?Mining?Industry?Herald，2000，4.
(11)VaAsquez?M，Espejo?R?T.Chemolithotrophic?bacteria?in?copper?oresleached?at?high?sulfuric?acid?concentration.Appl?Environ?Microbiol，1997，63：332-336.
(12)Espejo?R?T，Romero?J.Bacterial?community?in?copper?sulfide?oresinoculated?and?leached?with?solution?from?commercial-scale?copper?leachingplant.Appl?Environ?Microbiol，1997，63：1344-1352.
(13)Fowler?T?A，Crundwell?F?K.Leaching?of?zinc?sulfide?by?Thiobacillusferrooxidans?experiments?with?a?controlled?redox?potential?indicate?no?directbacterial?mechanism.Appl?Environ?Microbiol，1998，64：3570-3575.
(14)Karavaiko?G?I.Microbiological?processes?for?the?leaching?of?metalsfrom?ores.State-of-the-art?review.In：Torma?A?E，editor.Moscow：Centre?ofInternational?Projects，GKNT，1985.
Summary of the invention:
Realgar has been successfully used to the treatment of various skin disease clinically, demonstrates good antibiotic, antivirus action.But because the defective of existing processing technique, use Realgar many with heavy dose dissolving after-filtration or adopt suspension treatment dermatosis clinically, exist use amount big, the content height of poisonous metal or arsenic, problem such as toxicity is big.More can not fully dissolve and reduced drug effect, thereby reduce the bacteriostasis of Realgar and the treating for skin disease effect owing to Realgar.
The present invention adopts disclosed technical finesse Realgar in the Chinese invention patent 2006102000675, makes realgar microorganism extracting liquid.This realgar microorganism extracting liquid can (Gamma Magnitude) have the obvious suppression effect to comprising escherichia coli, staphylococcus aureus, Candida albicans, bacillus subtilis, epidermis drug resistance staphylococcus, excrement enterobacteria, non-drug resistance staphylococcus epidermidis, wax shape staphylococcus epidermidis, pseudomonas aeruginosa, herpesvirus etc. in extremely low concentration range.Be converted into solvable state through the Realgar behind the microbial treatments by insoluble state, this greatly reduces the use amount and the toxicity of realgar microorganism extracting liquid, has improved the drug effect in the treatment dermatosis simultaneously greatly, has reached the target of high-efficiency low-toxicity.
In addition, because realgar microorganism extracting liquid is a kind of medicine of solubility, therefore can be separately or with the other medicines compatibility and add pharmaceutically acceptable adjuvant, be easy to make various preparations, comprise suppository, pill, granule, membrane, microcapsule, drop pill, aerosol, medicated wine, syrup, oral liquid, injection or injection powder injection etc., this is that the researcher in this field is understandable.
The specific embodiment:
Embodiment 1: the preparation of realgar microorganism extracting liquid
Handle Realgar according to disclosed mineral drug bacterial treatment method in the Chinese invention patent 2006102000675, embodiment is as follows: take by weighing the 0.5g Realgar and cross in the 250mL conical flask of 200 purpose powder after sterilization, add the 9K iron-free liquid culture medium 90mL after sterilizing, transfer pH to 1.75 with acid, after treating that pH is stable, the inoculation volume fraction is that 20% thiobacillus ferrooxidant and final concentration are 1.0g/L FeSO
4, shaken cultivation 30d under 30 ℃, 135r/min condition.Decompress filter is collected supernatant, regulates about leachate pH to 7, promptly gets realgar microorganism extracting liquid after 0.22 micron filter membrane degerming excessively.
Embodiment 2: the external bacteriostatic experiment of Realgar leachate
With escherichia coli, staphylococcus aureus, Candida albicans, the excrement enterobacteria, bacillus subtilis, epidermis drug resistance staphylococcus, the non-drug resistance staphylococcus of epidermis, Bacillus cercus, pseudomonas aeruginosa adopt beef-protein medium activation breeding, and 30 ℃ of shaken cultivation were used for bacteriostatic experiment after 24 hours.Beef-protein medium prescription: Carnis Bovis seu Bubali cream 3.0g, peptone 10.0g, sodium chloride 5.0g, distilled water 1000ml moist heat sterilization.
Leachate: arsenic content is the realgar microorganism extracting liquid of 424.4 μ g/ml.
Positive control: levofloxacin hydrochloride and sodium chloride injection (Chengdu Pharmacy stock Co., Ltd of buchu group), be mixed with the solution of 4 μ g/ml, 4 ℃ of preservations, standby.
(bacterial concentration is 10 with cultured bacterium liquid
6Cfu/ml) 200ul drips on the beef extract-peptone solid medium, pushes away on flat board with hairclipper and smears evenly; The Oxford cup of bacterium of will going out is placed on the flat board that coats bacterium, and each dull and stereotyped going up is placed 4 Oxford cups, guarantees that spacing is even; In an Oxford cup, add 200ul positive control liquid, and on flat board, carry out labelling; Along clockwise direction, the Realgar leachate that in all the other 3 Oxford cups, adds the 200ul variable concentrations successively; Flat board is put into 30 ℃ of constant incubators to be cultivated 24 hours; Measure antibacterial circle diameter and record, external fungistatic effect of realgar microorganism extracting liquid such as table 1 at the dull and stereotyped back side with ruler.
The external fungistatic effect of table 1. realgar microorganism extracting liquid (unit: mm)
| Strain | Positive control | Stock solution | Dilute 2 times | Dilute 4 times |
| Epidermis drug resistance staphylococcus | 18 | 20 | 16 | 11 |
| Candida albicans | 18 | 17 | 14 | 12 |
| Staphylococcus aureus | 19 | 19 | 14 | 11 |
| The excrement enterobacteria | 20 | 18 | 15 | 11 |
| Pseudomonas aeruginosa | 17 | 19 | 15 | 13 |
| Bacillus cercus | 17 | 20 | 16 | 11 |
| Escherichia coli | 18 | 16 | 13 | 9 |
| Bacillus subtilis | 21 | 19 | 15 | 11 |
| Non-drug resistance staphylococcus epidermidis | 22 | 20 | 18 | 13 |
Embodiment 3: the Realgar leachate is to the minimal inhibitory concentration determination experiment of several antibacterials
With escherichia coli, staphylococcus aureus, Candida albicans, excrement enterobacteria, bacillus subtilis, epidermis drug resistance staphylococcus, the non-drug resistance staphylococcus of epidermis, Bacillus cercus, pseudomonas aeruginosa adopt beef-protein medium activation breeding, and 30 ℃ of shaken cultivation are standby.Beef-protein medium prescription: Carnis Bovis seu Bubali cream 3.0g, peptone 10.0g, sodium chloride 5.0g, distilled water 1000ml moist heat sterilization.
Leachate: arsenic content is the realgar microorganism extracting liquid of 424.4 μ g/ml.
Positive control: levofloxacin hydrochloride and sodium chloride injection (Chengdu Pharmacy stock Co., Ltd of buchu group), be mixed with the solution of 4 μ g/ml, 4 ℃ of preservations, standby.
Get 10 sterile test tube, add 2ml fluid medium, i.e. beef extract-peptone fluid medium in every test tube; In No. 1 test tube, add 2ml Realgar leachate, therefrom take out after shaking up in No. 2 test tubes of 2ml mixed liquor adding, shake up the back and take out No. 3 test tubes of 2ml mixed liquor adding ..., and the like, up to No. 8 test tubes; No. 9 test tubes are as blank; No. 10 test tubes add the 2ml positive control solution as positive control in No. 10 test tubes; (bacterial concentration is 10 to add 0.1ml culture of bacteria bacterium liquid at last in every
6Cfu/ml.), place 30 ℃ of constant incubators to cultivate observed result after 24 hours: mixed liquor is clear and bright after the tube shaken, thinks that this test tube does not have bacterial growth, and medicinal liquid has bacteriostasis; If opposite test tube vibration back mixed liquor becomes muddy and shows that then bacterial growth is arranged, and medicinal liquid does not have bacteriostasis, and the result is as shown in table 2.
Table 2. Realgar leachate is to the minimal inhibitory concentration measurement result (unit is μ g/ml) of several antibacterials
| Antibacterial | MIC |
| Staphylococcus aureus | 53.05 |
| Enterococcus faecalis | 53.05 |
| Bacillus subtilis | 26.53 |
| The drug resistance staphylococcus epidermidis | 53.05 |
| Escherichia coli | 53.05 |
| Non-drug resistance staphylococcus epidermidis | 26.53 |
| Bacillus cercus | 26.53 |
| Candida albicans | 106.1 |
| Pseudomonas aeruginosa | 26.53 |
Embodiment 4: the herpes zoster efficacy experiment
Herpes zoster is the viral dermatosis that varicella zoster virus causes, this is organized 10 cases and all adopts semar technique treatment, and the Realgar leachate of getting after the dilution is applied in the skin lesion place, one day three times in right amount.
Table 3. Realgar leachate is to the herpes zoster curative effect
Claims (3)
1. a realgar microorganism extracting liquid is characterized in that this realgar microorganism extracting liquid can suppress pathogenetic bacteria and virus, can be used to prepare antibacterials and antiviral drugs.
2. a kind of realgar microorganism extracting liquid according to claim 1 is characterized in that this leachate is the leachate of microbial treatments gained, prepares according to Chinese invention patent application 2006102000675 disclosed methods.
3. according to the described a kind of realgar microorganism extracting liquid of claim 1～2, it is characterized in that this realgar microorganism extracting liquid can be separately or with the other medicines compatibility and add pharmaceutically that the acceptable adjuvant is prepared into suppository, tablet, pill, granule, membrane, microcapsule, drop pill, aerosol, medicated wine, syrup, oral liquid, injection or injection powder injection.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108236613A (en) * | 2016-12-27 | 2018-07-03 | 兰州大学 | Realgar microorganism extracting liquid is preparing the application in inhibiting angiogenesis drug |
| CN111012799A (en) * | 2018-10-09 | 2020-04-17 | 兰州大学 | Application of realgar microbial leaching as PML-RAR α protein inhibitor |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1839880A (en) * | 2005-02-14 | 2006-10-04 | 兰州大学 | Heavy metal removal method from mineral drug |
-
2007
- 2007-10-16 CN CNA2007102020851A patent/CN101164550A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1839880A (en) * | 2005-02-14 | 2006-10-04 | 兰州大学 | Heavy metal removal method from mineral drug |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108236613A (en) * | 2016-12-27 | 2018-07-03 | 兰州大学 | Realgar microorganism extracting liquid is preparing the application in inhibiting angiogenesis drug |
| CN111012799A (en) * | 2018-10-09 | 2020-04-17 | 兰州大学 | Application of realgar microbial leaching as PML-RAR α protein inhibitor |
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Application publication date: 20080423 |