CN1290854C - Gentamicin, preparation method and uses thereof - Google Patents
Gentamicin, preparation method and uses thereof Download PDFInfo
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- CN1290854C CN1290854C CN 200410039138 CN200410039138A CN1290854C CN 1290854 C CN1290854 C CN 1290854C CN 200410039138 CN200410039138 CN 200410039138 CN 200410039138 A CN200410039138 A CN 200410039138A CN 1290854 C CN1290854 C CN 1290854C
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Abstract
The present invention relates to new gentamicin and a preparation method and purposes thereof. The preparation method is characterized in that glutamic acid or sodium glutamate or glutamic amine is added to a conventional micromonospora purpurea culture medium containing corn powder, starch, yeast powder or soybean powder, potassium nitrate, sodium carbonate, ammonium sulfate, peptone and cobalt chloride. A detailed formula and a detailed fermentation method are disclosed in the specification. The present invention has the advantages of high gentamicin C1 content and little toxic or side effect of C1, and the gentamicin of the present invention belongs to new gentamicin with good components. The products of the present invention are widely used for being used as antibiotics for medical care and animal husbandry.
Description
Technical field
The present invention relates to antibiotic fermentation, specifically relate to gentamicin of new generation, preparation method and its usage that fermentative preparation improves high-quality component content.
Background technology
Gentamicin is the microbiotic of finding in the U.S. in 1963.Worldwide be widely used in clinical.China began to produce gentamicin in 1967.China has become main in the world gentamicin producing country at present, and becomes maximum in the world gentamicin supply place.
Gentamicin is the microbiotic of being produced by deep red micromonospora (Micromonospora purpurea).It is the aminoglycosides microbiotic of component more than, and its molecule is made up of three ring texturees, is respectively osamine, 2-deoxystreptamine and deep red brown sugar amine.Owing on deep red brown sugar amine, have methylating of different positions, so gentamicin is made up of the methylated component of difference.Quality to gentamicin has clearly regulation in the world, and its standard mainly is to decide according to the ratio of gentamicin component, should be not less than 25% as gentamicinC1, and C1a should be controlled at 15-40%, and C2 and C2a should be controlled at 20-50%.
Over nearly 40 years, gentamicin is widely used in treating human diseases and livestock industry.Gentamicin is especially to Pseudomonas aeruginosa, and it is evident in efficacy that intestinal bacilli and golden Portugal bacterium infect, and improved people's health quality greatly, and guaranteed the stably manufactured of animal husbandry.But gentamicin is the same with some other medicine microbiotic, when the curative effect of demonstration is arranged, has supervened severe side effect.These toxicity mainly are reflected on the organs such as ear and kidney.
Scientist has carried out secular discussion to the toxicity that how to reduce gentamicin.These mainly concentrate on the preparation aspect, promptly add additive, as deferoxamine mesylate (Chen Yang etc.: the 34th the 3rd phase of volume of " Chinese hals,Nasen und Ohrenheilkunde magazine " June in 1999).But for various reasons, additive does not obtain promoting.At present, the side effect of gentamicin is not also improved at all.
The key of producing the hypotoxic gentamicin of high quality is how from improving the component of gentamicin basically.Because the different components in the gentamicin, its drug effect difference, and also toxicity is also different.As activity, then be that component C 1 is greater than C2 to faecalis and dysentery bacterium.But the median protective dose of C1 component (PD 50) is lower than other component.So the therapeutic index height of C1 component, good effect, and also toxic side effect is low.Therefore, produce high-quality gentamicin, the production technique that promptly improves the C1 component has also become the important topic of gentamicin industry.So, the invention provides the high-load gentamicin simple method for preparing of C1.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art part, and a kind of C1 content height, gentamicin of new generation, preparation method and its usage that toxic side effect is little are provided.
The object of the invention can realize by following measure: gentamicin component C 1 content 51-75% of new generation, Cla content 5-20%, C2 and C2a total content 15-35%; The preparation method: contain L-glutamic acid or Sodium Glutamate or glutamine in the fermention medium of routine, effective concentration is respectively the 0.1-50 grams per liter.According to a conventional method, pH6.8-7.6, culture temperature 35-37 ℃ fermented 88-92 hour.
It is Semen Maydis powder 5-15 grams per liter that substratum is formed, starch 10-50 grams per liter, yeast powder or analysis for soybean powder 25-40 grams per liter, saltpetre 0.01-0.45 grams per liter, lime carbonate 2.5-7.5 grams per liter, ammonium sulfate 0.1-3.5 grams per liter, peptone 3.5-7.5 grams per liter, cobalt chloride 5-50 mg/litre.
The bacterial classification that uses among the present invention is deep red micromonospora (Micromonospora purpurea bacterial strain NRRL2953) ATCC15835, this bacterium is deposited in American type culture collection (AmericanType Culture Collection, P.O.Box 1549, Manassas, VA 20110).
Gentamicin is purified and is adopted ordinary method.
The gentamicin proximate analysis:
Use high pressure lipuid chromatography (HPLC) (HPLC) to analyze gentamicin component (Fig. 2).
In 4 main component C 1 of gentamicin, C1a, among C2 and the C2a, C1 toxic side effect minimum.The main foundation of traditional gentamicin standard of Shi Yonging is the definition [mark (Marquez) etc.: United States Patent (USP) 3,651,042 (1972 year)] of the early stage patent of invention of the U.S. to the gentamicin component in the world.Tradition gentamicin and gentamicin component content of new generation definition and measured value thereof are as shown in table 1.
Table 1: gentamicin of new generation and traditional gentamicin component are relatively
As can be seen from Table 1, the present invention is defined as the C1 of 51-75% to gentamicin component content of new generation, the C1a of 5-20%, and the C2 of 15-35% and C2a significantly are different from the definition of traditional gentamicin component content.Especially gentamicinC1 content of new generation is higher than traditional gentamicin content, and traditional gentamicinC1 content is 25-50%, and the measured value of gentamicin component C 1 content of new generation that the present invention obtains is respectively 66.8%, 64.6% and 57.1%.
The present invention has following advantage compared to existing technology: the present invention adopts fermentation process, prepared gentamicinC1 content 51-75%, and than traditional C1 content 25-50% height, the C1 poison is secondary with little, is the gentamicin that there is high-content high-quality component in a new generation.
Description of drawings:
Fig. 1: gentamicin molecular structure
Be depicted as the structure difference of the main component of gentamicin, its difference is the methylating in various degree of deep red brown sugar amine.
R1 among the figure, R2 and R3 represent hydrogen atom or methyl respectively.
GentamicinC1: R1 and R2 are methyl; R3 is a hydrogen atom.
Gentamicin C1a: R1, R2 and R3 are all hydrogen atom.
GentamicinC2: R1 and R3 are hydrogen atom; R2 is a methyl.
GentamicinC2 a:R1 and R2 are hydrogen atom; R3 is a methyl.
Fig. 2: the HPLC analytical results of gentamicin component of the present invention
X-axis is the elution time of gentamicin among the figure, and unit is minute; Y-axis is the absorbancy in 254 nanometers, and unit is a milli absorbance unit (mA).
Fig. 3: the HPLC analytical results of traditional technology gentamicin component
As can be seen from Figures 2 and 3, the gentamicin of zymotechnique preparation of the present invention, the gentamicin that the content of its C1 is produced apparently higher than traditional technology.
Specific implementation method:
Enumerate four embodiment below, the present invention is further specified.But the present invention is not only limited to these embodiment.
Embodiment 1: gentamicinC1 high yield cultural method.Contain Semen Maydis powder 11 grams per liters, starch 25 grams per liters, analysis for soybean powder 32 grams per liters, saltpetre 0.01 grams per liter, lime carbonate 5 grams per liters, ammonium sulfate 0.25 grams per liter, peptone 4.5 grams per liters are in the conventional fermention medium of the deep red micromonospora of cobalt chloride 15 mg/litre, add L-glutamic acid 2.5 grams per liters, pH7.2,35 ℃ of culture temperature, fermentation time 90 hours.
Embodiment 2: in the fermention medium identical with embodiment 1, contain Sodium Glutamate 2.5 grams per liters, replace L-glutamic acid among the embodiment 1, all the other are all identical with embodiment 1.
Embodiment 3: in the fermention medium identical with embodiment 1, contain glutamine 2.5 grams per liters, replace the L-glutamic acid among the embodiment 1, all the other are all identical with embodiment 1.
The measured value of embodiment 1-3 sees Table 1.
Embodiment 4: the product of the embodiment of the invention 3 is contained C157.1%, and the gentamicinC1 content 35.4% with conventional production methods is produced mixes by 1: 1 weight ratio, promptly obtains C1 content and be 46.25% gentamicin product.
The purifying of gentamicin.In the nutrient solution of embodiment 1-3, add sulfuric acid, make pH reduce to 2-3, and kept 30-60 minute, afterwards, add sodium hydroxide, pH is heightened 6.0-6.5.Add strong cation-exchanging resin (Dowex-50), fully adsorb gentamicin.Through ammonia chloride (0.4 mole) and lower concentration ammoniacal liquor (0.1 mole) flushing, use high density ammoniacal liquor (1 mole) to dissociate at last.
The gentamicin proximate analysis.Use high pressure lipuid chromatography (HPLC) (HPLC) to analyze the gentamicin component.Analytical procedure is seen 1988 19 volumes of Qiao Ke " chromatogram magazine " (Journal of Chromatography) such as (Jork) 115-9 page or leaf.
Illustrate a bit: international standard gentamicinC1 content 25-50%, in fact the gentamicinC1 content about 35% that makes of production unit does not reach C1 content 36-50% scope.And gentamicin of the present invention mixes by weight proportion with traditional gentamicin, can make to contain C1 near 50% international standard gentamicin.
Gentamicin of the present invention is widely used in medical treatment and livestock industry, makes microbiotic.
Claims (3)
1, gentamicin of new generation is characterized in that gentamicin component C 1 content 51-75%, Cla content 5-20%, C2 and C2a total content 15-35%.
2, the preparation method of gentamicin of new generation as claimed in claim 1 is characterized in that: in the fermention medium of routine, contain L-glutamic acid or Sodium Glutamate or glutamine, effective concentration is respectively the 0.1-50 grams per liter.
The purposes of 3 gentamicins of new generation as claimed in claim 1 is characterized in that: it is used for mixing by weight proportion with the gentamicin of produced in conventional processes, can make to contain the gentamicin of C1 near 50% international standard.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200410039138 CN1290854C (en) | 2004-02-12 | 2004-02-12 | Gentamicin, preparation method and uses thereof |
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| CN 200410039138 CN1290854C (en) | 2004-02-12 | 2004-02-12 | Gentamicin, preparation method and uses thereof |
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| CN1557958A CN1557958A (en) | 2004-12-29 |
| CN1290854C true CN1290854C (en) | 2006-12-20 |
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| CN 200410039138 Expired - Fee Related CN1290854C (en) | 2004-02-12 | 2004-02-12 | Gentamicin, preparation method and uses thereof |
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102558255B (en) * | 2010-12-24 | 2016-01-20 | 无锡济民可信山禾药业股份有限公司 | A kind of glucoside-containing component and extraction and separation method thereof |
| US20130075085A1 (en) * | 2011-09-23 | 2013-03-28 | E. I. Du Pont De Nemours And Company | Use of glutamate for microbial enhanced oil recovery |
| CN102363759B (en) * | 2011-10-27 | 2012-10-17 | 福州大学 | Engineering bacteria for producing gentamicin C1a and application thereof |
| US10555961B2 (en) | 2015-06-05 | 2020-02-11 | Indiana University Research And Technology Corporation | Materials and methods for treating bacterial infections using C-1 gentamicin |
| CN105503973B (en) * | 2015-12-25 | 2018-06-08 | 无锡济民可信山禾药业股份有限公司 | A kind of glucoside-containing component and its extraction separation method |
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Granted publication date: 20061220 Termination date: 20140212 |