CN101264103B - Application of native copper native copper microbial extracting liquid in preparing medicaments for treating bone fracture - Google Patents
Application of native copper native copper microbial extracting liquid in preparing medicaments for treating bone fracture Download PDFInfo
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- CN101264103B CN101264103B CN2008103011144A CN200810301114A CN101264103B CN 101264103 B CN101264103 B CN 101264103B CN 2008103011144 A CN2008103011144 A CN 2008103011144A CN 200810301114 A CN200810301114 A CN 200810301114A CN 101264103 B CN101264103 B CN 101264103B
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Abstract
The invention relates to the application of microbial leaching solution of natural-copper in preparing drugs treating fracture. The leaching solution of natural-copper is prepared according to the bacteria processing method publicized by 2006102000675 of Chinese invention patent application. The microbial leaching solution of natural-copper has the advantages that: leaching solution of natural-copper has obvious result in promoting healing of the fracture with small amount; because leaching solution of natural-copper is soluble and can be easily made into each formulation singly or jointly with other drug compatibility or accessories accepted pharmaceutically. The formulation has a plurality types comprising suppository, medical pill, granule, film agent, microcapsules, dropping pill, aerosol preparation, medicinal wine, syrup, oral liquid/solution, injection or injection of injection powder and other formulations.
Description
Technical field
The present invention relates to the application of Pyritum microorganism leachate in preparation treatment fracture medicine; Specifically utilize microbial technique; The dissolving of Chinese medicine Pyritum is leached; Then Pyritum microorganism leachate is used to prepare the technology of treatment fracture medicine, belongs to Chinese medicine new technique, new technology field.
Background technology
Pyritum (PYRITUM) is a sulfide-based mineral yellow iron mine family pyrite, the main ferrous disulfide (FeS that contains
2), be common minerals as Chinese medicine.Pyritum has effects such as dissipating blood stasis, synthetism, pain relieving, is used for tumbling and swelling, muscles and bones folding wound
[1]Modern medicine thinks that Pyritum has the effect that promotes union of fracture, on clinical and pharmacological research, has been successfully used to the treatment of union of fracture, therefore more and more receives the extensive concern of Chinese scholars, becomes a focus of mineral Chinese medicine research.
Mineral drug is the culture rarity of the Chinese nation, but because its water-insoluble, causes problems such as low, the clinical consumption of bioavailability is big, toxicity height, the development that has seriously restricted mineral Chinese medicine with crack the international market.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 meaning very great.
The main method that is used at present concoct Pyritum is for forging the method for quenching, and still there are many problems in this method
[2-4], at first, not only time-consuming, effort, power consumption, and in the calcine process, can produce a large amount of sulfur dioxide gas, serious environment pollution influences health of operators; Secondly, forging the method for quenching can not effectively dissolve Pyritum, makes its use receive some restrictions, more can't quantitatively control active constituent content in the medicine, is difficult in the injectable drug prescription, use etc.
Theoretically, adopt chemical method can dissolve Pyritum, but chemical method require harsh conditions such as nitric acid, hydrochloric acid, perchloric acid, is difficult to preserve necessary effective ingredient in the Pyritum, thereby loses the therapeutical effect of Pyritum; On the other hand, adopt chemical method to handle the composition that Pyritum also possibly not had medicine itself, even with poisonous and hypertoxic composition introducing medicine.
Because above-mentioned all variety of problems that is difficult to overcome, cause on the current market Pyritum quality very different, Pyritum and Related product thereof are difficult to move towards the international market.
At present, a high-new biotechnology microbial metallurgy technology is a dark horse
[5-7]Under certain air, pH, temperature conditions, the Ore lysate that some special antibacterials (thiobacillus ferrooxidant, thiobacillus thiooxidans etc.) can be changed insoluble sulfide into solubility reclaims required element then
[8-10]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.Pyritum also is a kind of sulphide ore, has by the dissolved potential of leaching microbacteria.
Chinese invention patent 2006102000675 discloses the microbial treatment method of mineral Chinese medicine
[11]The present invention relates to the application of Pyritum leachate in preparation treatment fracture medicine of microbial treatments, this leachate can play good therapeutical effect to union of fracture.Because through the dissolving and the conversion of microorganism, the plurality of active ingredients in the Pyritum is effectively dissolved, so this Pyritum microorganism leachate can be brought into play effect in low strength range.
List of references:
[1] Pharmacopoeia of the People's Republic of China version in 2005 is one one.
[2] analogy qin cloud, Xu Jinhua.The Pyritum processing procedure is inquired into.China's Pharmaceutical, 2002,11 (10): 64
[3] Lou Guanfeng.The improved discussion of Pyritum concocting method.The time precious traditional Chinese medical science traditional Chinese medicines, 2003,14 (8): 457
[4] Yan Hui, Zhang Li, Ding Anwei.The process of preparing Chinese medicine mechanism research of Pyritum.The time precious traditional Chinese medical science traditional Chinese medicines, 2004,15 (11): 757
[5]Torma?A?E.The?role?of?Thiobacillus?ferrooxidans?in?hydrometallurgialprocesses.?Adv?Biochem?Eng.1977，6：1
[6]Colmer?A?R，Hinckel.M?E.The?role?of?microorganism?in?acid?mine?drainage.A?preliminary?report，Science，1947，106：253.
[7]Vaquez?M，Espejo?R?T.Chemolithotrophic?bacteria?in?copper?ores?leachedat?high?sulfuric?acid?concentration.Appl?Environ?Microbiol，1997，63：332.
[8]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.
[9]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.
[10]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.
[11] Li Hongyu, Zhang Jinghong, Zhi Dejuan.200610200067.5, the heavy metal removal method in the mineral drug.
Summary of the invention:
The method for preparing of Pyritum microorganism leachate: take by weighing the 0.5g Pyritum; Grind 200 mesh sieves, put into clean 250mL conical flask, added 9 K iron-free liquid culture medium 100mL after sterilizing; 5M sulphuric acid is transferred pH to 1.60; After treating that pH is stable, the inoculation volume fraction is 10% thiobacillus ferrooxidant, and blank is not inoculated bacterium liquid.After taking by weighing quality, leached 15 days at 25 ℃, 130r/min.Adopt 5M sulphuric acid to regulate pH value in the experimentation, replenish institute's moisture evaporated with distilled water.Shake the bottle leaching and stop after 15 days, hold over night is collected supernatant, and is about regulator solution pH to 7, after 0.22 micron filter membrane degerming of mistake, subsequent use.
Fracture rat oral Pyritum leachate is after 21 days, and alkaline phosphatase activities obviously improves, and this Pyritum leachate can promote the fracture growth of spur, explains that this Pyritum leachate has the effect that promotes union of fracture.
The same domain research worker is appreciated that; Handle the Pyritum microorganism leachate of gained through adopting said method; Can process various preparations; Comprise: suppository, suppository, tablet, pill, granule, membrane, microcapsule, drop pill, aerosol, medicated wine, syrup, oral liquid, injection or injection powder injection, these pharmaceutical compositions can contain 0.1%～100% Pyritum microorganism leachate.
The present invention has overcome the defective of prior art, and Pyritum leachate use amount provided by the present invention has reached safe medication well below traditional Pyritum, and purpose evident in efficacy.
The specific embodiment:
The preparation of embodiment one Pyritum microorganism leachate
Take by weighing the 0.5g Pyritum, ground 200 mesh sieves, get powder.In clean 250mL conical flask, add the 9K iron-free liquid culture medium 100mL after the sterilization, 5M sulphuric acid is transferred pH to 1.60, treat that pH is stable after, the inoculation volume fraction is 10% thiobacillus ferrooxidant, blank is not inoculated bacterium liquid.After taking by weighing quality, leached 15 days at 25 ℃, 130r/min.Adopt 5M sulphuric acid to regulate pH value in the experimentation, replenish institute's moisture evaporated with distilled water.Leaching stopped flask for 15 days, allowed to stand overnight, the supernatant was collected, to adjust the solution pH to about 7, over 0.22 Jing filter sterilization, the standby.
Embodiment two Pyritum leachates are to the influence of fracture rat blood serum calcium concentration and alkaline phosphatase activities
Laboratory animal: the healthy SD rat, female, 6～8 ages in week, body weight (200 ± 20) g, the SPF level is purchased in Lanzhou University zoopery center.
Fracture model: pentobarbital sodium anesthesia SD rat, the right hind depilation, iodophor disinfection, the scalpel vertical cut is opened skin, exposes tibia, and is not exclusively sawed-off in last 1/3 place's cut-out sclerotin (about 1mm is dark, 2mm is wide), keeps connecting, and skin suture, postoperative are unfixing.Postoperative intramuscular injection penicillin, 40,000 units/only, intramuscular injection three days.Postoperative the 4th day beginning gastric infusion, the fracture rat is divided into three groups: positive group (n=9, bone setting tablet is dissolved in distilled water after pulverizing, 12/d of people's consumption proportionately, the conversion rat is 1.305/kg.d, i.e. 522mg/kg.d); Experimental group (n=9, Pyritum leachate 20ml/kg.d is equivalent to Pyritum 44mg/kg.d); Negative group (n=9, distilled water 20ml/kg.d).
Every treated animal is respectively at postoperative 7d, 14d, the blood sampling of 21d heart, and 4 ℃ left standstill 4 hours, and the centrifugal 10min of 2000rpm gets supernatant.Automatic clinical chemistry analyzer is measured Serum alkaline phosphatase activity.
The mensuration result of serum alkaline phosphatase shows that the experimental group Serum alkaline phosphatase activity reaches peak at 14d, begins then to reduce, and returns to normal value.When 14d, the experimental group Serum alkaline phosphatase activity is organized apparently higher than feminine gender, and illustrative experiment group medicine has certain therapeutical effect to the rat fracture.
Table 1 leachate is to the influence (U/L) of fracture rat blood serum alkali phosphatase
Annotate: * compares P＜0.05 with negative group
Embodiment three Pyritum leachates are to the influence of fracture rat fracture growth of spur
Fracture model is with embodiment two.Every treated animal carries out X-ray respectively at postoperative 7d, 14d, 21d and takes.Readding sheet evaluates according to the Pyatyi evaluation criteria of union of fracture:
The I level: broken ends of fractured bone edge is fuzzy slightly.Periosteal reaction is slight, no callus.
The II level: broken ends of fractured bone edge is obviously fuzzy, and interior external callus density is higher than soft tissue, and callus appears in fracture gap more than 1/2.
The III level: broken ends of fractured bone edge is most of to disappear, and interior external callus is similar to pulp cavity, and fracture gap 1/2 above callus connects.
The IV level: broken ends of fractured bone edge is tending towards disappearing, and interior external callus is similar to cortical bone, and fracture gap 1/2 above callus connects.
The V level: the complete obiteration of broken ends of fractured bone edge, the same cortical bone of interior external callus, and connection, moulding, pulp cavity are logical again.
The X-ray sheet shows that administration group rat fracture bony callus area is obviously greater than the area (table 2) of blank group rat fracture callus.
Table 2 is respectively organized the X ray examination result of union of fracture degree
Claims (1)
1. the method for preparing of a Pyritum microorganism leachate, this method comprises: take by weighing the 0.5g Pyritum, ground 200 mesh sieves; Get powder, put into clean 250ml conical flask, add the 100ml fluid medium after sterilizing; 5M sulphuric acid is transferred pH to 1.60, and after pH was stable, the inoculation volume fraction was 10% thiobacillus ferrooxidant; Leach 15d under 25 ℃, 130r/min condition, adopt 5M sulphuric acid to regulate pH value in the experimentation, replenish institute's moisture evaporated with distilled water; Shake the bottle leaching and stop after 15 days, hold over night is collected supernatant; Regulate about leachate pH to 7, promptly get Pyritum microorganism leachate after 0.22 micron filter membrane degerming excessively, it is characterized in that: described fluid medium is a 9K iron-free liquid culture medium.
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| CN2008103011144A CN101264103B (en) | 2008-04-14 | 2008-04-14 | Application of native copper native copper microbial extracting liquid in preparing medicaments for treating bone fracture |
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| CN2008103011144A CN101264103B (en) | 2008-04-14 | 2008-04-14 | Application of native copper native copper microbial extracting liquid in preparing medicaments for treating bone fracture |
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| CN101264103B true CN101264103B (en) | 2012-08-22 |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109528765A (en) * | 2018-11-12 | 2019-03-29 | 兰州大学 | Native copper leachate is in cultured osteoblast-like cells in vitro proliferation and the purposes in mature mineralising treatment |
| CN109609447A (en) * | 2019-02-18 | 2019-04-12 | 兰州大学 | The preparation of native copper leachate promotes the application of mesenchymal stem cells differentiation drug |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1089147A (en) * | 1993-01-01 | 1994-07-13 | 李海涛 | The pure Chinese medicinal preparation and the concocting method of treatment fracture |
| CN1839880A (en) * | 2005-02-14 | 2006-10-04 | 兰州大学 | Heavy metal removal method from mineral drug |
| CN1907410A (en) * | 2006-08-02 | 2007-02-07 | 朴喜新 | Chinese traditional medicine preparation for treating bone fracture and traumatic injury |
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2008
- 2008-04-14 CN CN2008103011144A patent/CN101264103B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1089147A (en) * | 1993-01-01 | 1994-07-13 | 李海涛 | The pure Chinese medicinal preparation and the concocting method of treatment fracture |
| CN1839880A (en) * | 2005-02-14 | 2006-10-04 | 兰州大学 | Heavy metal removal method from mineral drug |
| CN1907410A (en) * | 2006-08-02 | 2007-02-07 | 朴喜新 | Chinese traditional medicine preparation for treating bone fracture and traumatic injury |
Non-Patent Citations (1)
| Title |
|---|
| 庞仁俊.自然铜浸取及其浸出液的药效研究.《兰州大学研究生学位论文》.2007,摘要. * |
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