CN1027905C - Preparation method of antiparasitic new antibiotic and its salts - Google Patents
Preparation method of antiparasitic new antibiotic and its salts Download PDFInfo
- Publication number
- CN1027905C CN1027905C CN 88100187 CN88100187A CN1027905C CN 1027905 C CN1027905 C CN 1027905C CN 88100187 CN88100187 CN 88100187 CN 88100187 A CN88100187 A CN 88100187A CN 1027905 C CN1027905 C CN 1027905C
- Authority
- CN
- China
- Prior art keywords
- mycin
- platform
- antiparasitic
- acid
- salts
- 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.)
- Expired - Fee Related
Links
Images
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
Abstract
The present invention relates to a preparation method of antiparasitic new antibiotic and salts, which is characterized in that two kinds of substances with antiparasitic activity are separated, extracted and refined from KP-197 strain culture by means of a solvent method. The antiparasitic activity of the substances is obviously higher than that of known medicines, but the toxicity of the substances is lower than that of the known medicines. Therefore, the present invention can become an effective medicine for treating human, animal and plant parasites.
Description
The present invention relates to a kind of manufacture method that can from microorganisms cultures, extract antiparasitic new antibiotics ring platform mycin (JEITACINS) and basic salt.This microbiotic and the chemical structural formula that can make medicinal salt thereof are:
In the formula: R=sec.-propyl or isobutyl-
Know, in microbiotic, Ao Fomai rhzomorph (AVERMECTIN), paromycin (PAROMOMYCIN) are arranged, in synthetic drug, LEVAMISOLE HCL (LEVAMISOLE), niclosamide (NICLOSAMIDE) etc. are arranged as antiparasitic medicine.It is sure that these medicines are used to treat its effects such as oxyuriasis, filaricide as pharmaceuticals and animal drugs.But known anti-parasite medicine has problems such as toxicity, side effect or resistance.
Purpose of the present invention is to seek the littler anti-parasite medicine of toxic side effect, in the hope of providing parasiticide active material, solves the demand in the humans and animals treatment.
Main points of the present invention are to be target to explore new biologically active substance, separate bacterial classification from different pedotheques, and its product is studied.Found that the KP-197 bacterial strain, in its culture, contained the material that suppresses the parasite locomotor activity.From the culture of this bacterial strain, separate and make with extra care out the pure substances of two inhibition parasite locomotor activities.Study the physicochemical property of these two materials, measured its chemical structure.Material with this chemical structure is not seen report as yet.These materials are referred to as guards against platform mycin (JEITACINS).R is that the compound of sec.-propyl is guarding against platform mycin A(JEITACINSA in the structural formula), R is that the compound of isobutyl-is ring platform mycin B(JEITACINSB).The physico-chemical property of guarding against platform mycin A and ring platform mycin B is as follows:
Guard against platform mycin A:
(1) white powder
(2) ultimate analysis
C
18H
34N
2O
2
Calculated value C69.57; H11.04; N9.02%
Experimental value C70.40; H11.09; N7.68%
(3) molecular weight and molecular formula
Guard against the mass spectrum of platform mycin A, (m+H) arranged at the m/z311 place
+Molecular ion peak; In high resolution mass spec is analyzed, (M-OH) arranged at the m/z293 place
+Quasi-molecular ions.Determine its molecular formula C thus
18H
34N
2O
2, molecular weight is 310.
(4) ultra-violet absorption spectrum
As shown in Figure 1, a distinctive absorption peak is arranged at the 228mm place; An acromion appears at the 250nm place.
(5) infrared absorption spectrum
2950,2870,1705,1475,1415,1380,1270,1090,960(cm measure with the KBr solvent method, as shown in Figure 2, it is as follows that it mainly absorbs very big wave number:
-1).
(6) nuclear magnetic resonance spectrum
Use Valley peace XL-400, the 400MH nuclear magnetic resonance analyser is measured in heavy chloroform
1The H nuclear magnetic resonance spectrum as shown in Figure 3.
(7) color reaction
Aubepine-sulfuric acid the positive
The potassium permanganate positive
The sulfuric acid feminine gender
The Dragendorff test feminine gender
(8) solvability
Be dissolved in acetone, ethyl acetate, chloroform, hexane, dimethyl sulfoxide (DMSO).Be insoluble to methyl alcohol, water.
(9) chemical structure
In the structural formula that above-mentioned technical field is illustrated, R is the structural formula of sec.-propyl, and basis listed physico-chemical property in (1)-(9) and known similar compound compare and obtain, and it is 14-methyl isophthalic acid-ethene azo oxygen 15-alkane-8 ketone.
Guard against platform mycin B:
(1) colorless oil
(2) molecular formula in high resolution mass spectrum is analyzed, has (M-N) quasi-molecular ions in the m/z307 place, determined that thus its molecular formula is C
19H
36N
2O
2, molecular weight is 324.
(3) ultra-violet absorption spectrum
With guarding against platform mycin A
(4) infrared absorption spectrum
With guarding against platform mycin A
(5) color reaction
With guarding against platform mycin A
(6) solvability
Substratum growth substrate mycelium aerial hyphae soluble pigment
Yeast-malt extract is good, projection, and the oldlace light amber does not have brown
(3EC) (3IC) (4NE) for agar (ISP)
Rolled oats agar is medium to be soaked into, shallow wheat straw look shallow wheat straw look nothing
（ISP） （ZEA） （ZEA）
Inorganic salt starch fine jade is good, and light amber bamboo look poorness is white rare, the pearl pink colour
(3IC) (2GC) look (A) is (3CA) for fat (ISP)
Glycerine-arginine is medium, oldlace oldlace light tan
Agar (ISP) (3EC) (3EC) does not have (3GC)
Glucose-smart ammonia poorness, colour of camel's hair colour of camel's hair nothing
Acid agar (ISP) is (3IE) (3IE)
Peptone-yeast poorness, castor look castor look does not have the castor look
(3LI) (3LI) (3LI) for cream agar (ISP)
Tyrosine agar is medium, colour of camel's hair colour of camel's hair poorness, Sparrow Faeces brown
(3IE) (3IE) look (A) (3NI)
Sucrose-nitrate poorness, the shallow oyster white nothing of shallow oyster white
Agar (2CA) (2CA)
Glucose-nitric acid is medium, and the fallow nothing of light yellowish brown does not have
Salt agar (3LE) (3LE)
Glycerine-oxysuccinic acid poorness, the bamboo look is to the very poor bamboo look-golden brown of golden brown bamboo-Jin
Brown (2GC
Calcium agar (2GC-3PG)-3PG) white (A) (2GC-3PG)
Glucose-albumen is medium, projection, and wrinkle, the brown no light brown-oak of shallow spices is brown
Peptone agar oldlace (2CA) look (4LG) look (4LE-4PI)
The nutrient agar medium poorness, the shallow oyster white nothing of shallow oyster white
（2CA） （2CA）
With guarding against platform mycin A
(7) chemical structure
Guarding against platform mycin B is the analogue of guarding against platform mycin A, both charcoal atomicity differences on alkyl group side chain.
In the structural formula that above-mentioned technical field is illustrated, R is the structural formula of isobutyl-, is to compare the result who obtains according to listed physico-chemical property in (1)-(7) and known compound, and it is 15-methyl isophthalic acid-ethene azo oxygen base 16 alkane-8 ketone.
The microorganism that can produce guard against the platform mycin belongs to streptomyces, and the KP-197 bacterial strain of the streptomyces that for example is separated in the present invention is one of effective strain that uses in the present invention.This bacterial strain is kept at China Committee for Culture Collection of Microorganisms common micro-organisms center with the title of Streptomyces sp.KP-197, is called for short CGMCC, numbering CGMCC, NO.0130, preservation date on September 1st, 1987.The systematics property of this bacterial classification is as follows:
(1) the mycelia fracture is not observed in the vegetative hyphae prosperity on various substratum.On the starch minimal medium, aerial hyphae is very rare, white, and microscopic examination is found, fibrillae of spores is a linearity, and spore chain is made up of 20 above spores, and the spore size is 0.7-1.0 * 0.7 μ m, and is cylindrical, spore surface is smooth, does not find sclerotium, sporocyst and zoospore.
(2) feature on various substratum:
Study the cultural characteristic of this generation bacterium with Sai Gewa (313 pages of E.B.SHIRLING and D.G.GOTTLIEB-international system bacteriology magazine 16 volumes, 1966) method, it is as shown in the table for the result: (seeing the page 3 table)
(3) physiological property
Melanochrome forms the positive
The tyrosine agar positive
The peptone yeast extract paste iron agar positive
Glucose peptone gelatine culture (21-23 ℃) positive
The Tryptones yeast extract paste liquid nutrient medium positive
The tyrosine oxidase reacting positive
The generation feminine gender of hydrogen sulfide
The nitrate reduction positive
Gelatine liquefication (glucose, peptone, gelatine culture) (21-23 ℃) positive
The starch hydrolysis positive
Milk solidifies (37 ℃) feminine gender
Milk peptonizes (37 ℃) positive
Growth temperature range 15-38 ℃
The charcoal source utilizes (adopting Pu Gewa PRIDHOM-GOTTLIEB) agar as basic medium):
Utilize: D-glucose, D-fructose, seminose
Utilize slightly: D-N.F,USP MANNITOL, (pectinose, D-wood sugar, sucrose,
Do not utilize: L-rhamnosyl, I-inositol, raffinose, close disaccharides, Whitfield's ointment.
Mierocrystalline cellulose decomposes negative
(4) cell walls is formed
The diaminopimelic acid of cell walls (DAP) is LL-DAP.
In sum, the taxonomy feature of this bacterial strain can be summarized as follows: the diaminopimelic acid of cell walls is LL-DAP, and the shape of fibrillae of spores is a linear, spore chain, and spore surface is smooth, and vegetative hyphae is cream-coloured, and aerial hyphae is a white, forms melanochrome.Can determine that thus this bacterial strain belongs to a kind of of streptomyces (STREPTOMYCES).According to general Chai Shi classification (PRIDHAM AND TRESNER) (uncle states the 8th edition 748-829 page or leaf of watt division bacteria handbook, 1974), confirm that the bacterial strain of this generation belongs to a kind of of white series.
In the present invention, as the example of the microorganism in substratum, at first used to produce the microorganism that belongs to streptomyces of guarding against the platform mycin.Yet,, also can use in the present invention as long as can produce the change strain of this bacterial strain of guarding against the platform mycin.
As substratum, can use synthetic medium or natural medium.These substratum contain other nutritive substance that is suitable for cultivating actinomycetic carbon source, nitrogenous source and inorganic salts and an amount of necessity usually.
Carbon source in the substratum can be used various carbohydrate such as glucose, glycerine, fructose, maltose, seminose, wood sugar, semi-lactosi, ribose, starch and hydrolyzate thereof, and working concentration is generally 0.1-5%.Also have alcohol compounds such as amino acid such as organic acid, glycine, L-glutamic acid, L-Ala, methyl alcohol, ethanol such as gluconic acid, pyruvic acid, lactic acid, acetic acid in addition, and non-aromatic hydrocarbon polymer such as n-paraffin or vegeto-animal various lipid can use all.
As nitrogenous source, can use mineral acid or itrogenous organic substance matter such as organic acid ammonium salt, urea, peptone, NZ-amine, meat extract, yeast extract paste, dry yeast, corn steep liquor, casein hydrolysate, fish meal or its hydrolyzate, soyflour or its hydrolyzate, skimmed soy beans or its hydrolyzate such as ammonium, ammonium chloride, ammonium phosphate, ammonium sulfate, ammonium nitrate.Also have amino acid such as glycine, L-glutamic acid, L-Ala.Inorganic salts uses various phosphoric acid salt, vitriol, salt etc. and trace heavy metals salt.
When use has the variant of special nutrition requirement, must in substratum, add the material that can satisfy its nutritional requirement.Such nutritive substance when use contains the substratum of crude substance, must not add additional especially.
Fermentation culture is normally carried out under aerobic conditions such as vibration or aeration-agitation.Be to adopt the deep ventilation stir culture during actual production.The pH value of substratum can be 5-8, but generally better with neutral pH.Culture temperature can be 20-40 ℃, but generally 26-32 ℃ better (ideal temperature is about 27 ℃).The liquid culture time is generally 3-6 day, guards against tiring of platform mycin and stops when reaching the climax cultivating.Culture condition such as substratum composition, culture temperature, stirring velocity, air flow can suitably be adjusted selectedly according to the bacterial strain that uses and ambient conditions, can suitably select silicone resin, the plant wet goods tensio-active agent foam that disappears when foam appears in liquid culture for use.
Ring platform mycin with aforesaid method accumulates in culture is present in the nutrient solution usually.To guard against the platform mycin in order from culturing filtrate, extracting, generally to adopt the method for from microorganisms cultures, extracting meta-bolites.These methods can be separately or arbitrarily in conjunction with or use repeatedly.For example: filter, centrifugal, dialysis, concentrated, dry, freezing, absorption, wash-out; To the method for various solvent different solubilities (, adverse current molten as precipitation, crystallization, recrystallization, commentaries on classics joined cloth), methods such as chromatography.
Mainly be created in the nutrient solution owing to guard against the platform mycin,, at first from nutrient solution, remove thalline, and then from culturing filtrate, extract this material therefore in order to separate, extract this compound.
Guard against the platform mycin in order from culturing filtrate, to extract, use extractings such as non-hydrophilic organic solvent such as ethyl acetate, perhaps adsorb with gac, aluminum oxide, porousness high molecular synthetic resin, ion exchange resin etc., again with solvent wash-outs such as ethyl acetates, extracting solution that obtains or elutriant carry out concentrating under reduced pressure, again with extractions such as hexanes.The crude extract that obtains is like this made with extra care with method such as the silica gel or the alumina column chromatography etc. of usually refining lipid-soluble substance.
This compound can be made medicinal salt example hydrochloric acid salt, vitriol, nitrate, phosphoric acid salt, carbonate, tartrate, citric acid, succinate etc., uses ordinary method to prepare.
Advantage of the present invention and positively effect be, with the active substance of method for preparing, the existing medicine of its antiparasitic specific activity is strong, and its toxicity is lower than existing medicine.Concrete experiment is as treatment target with pine wilt nematode (BURSAPHLENCH US LIGNICLOUS), adopt the Kimura to wait method (the AGRICULTURE BIOLOGI CAI CHEMISRY 45 that introduces, 249,1981) measure the locomotor activity of guarding against platform mycin inhibition pine wilt nematode, consequently:
Guard against the lethality rate (%) of platform mycin A to nematode
0.5ug/ml 100%
0.25ug/ml 100%
0.125ug/ml 99%
0.063ug/ml 86%
Guard against the lethality rate (%) of platform mycin B to nematode
0.5ug/ml 100%
0.25ug/ml 100%
0.125ug/ml 93%
0.063ug/ml 80%
Known drug AVERMECTIN BLA is to the lethality rate (%) of nematode
5ug/ml 100%
2.5ug/ml 100%
1.25ug/ml 64%
0.63ug/ml 58%
To find out, the parasiticide specific activity known drug of guarding against the platform mycin is more than strong ten times from above comparison test, and the former obviously is better than the latter at curative effect.
Acute toxicity test be with mouse as laboratory animal, guard against platform mycin 10mg/kg through abdominal channels injection and do not cause death, observe the acute malicious shape of known medicine AVERMECTIN BLA, dead mouse as a result with same procedure.That as seen, guards against the platform mycin acutely is lower than known anti-parasite medicine.
Above activity and toxicity test show, the resulting ring platform of the present invention mycin is useful as the antiparasitic of people, animal and plant.
Embodiment:
100ml substratum (the substratum composition: glucose 0.1%, yam starch 0.4%, peptone 0.5% of in 500ml plate mouth bottle, packing into, meat extract 0.3%, yeast extract paste 0.5%, lime carbonate 0.4%, pH7.0), 121 ℃, behind 15 minutes steam sterilizings, slant culture (its composition: glycerine 1.0% with the KP-197 of a platinum loop, calcium malate 1.0%, yeast extract paste 0.1%, agar 1.5%) plant in the substratum of above-mentioned sterilization, on rotary shaker, cultivated three days for 27 ℃, should cultivate as seed.30 liters fermentor tank is two fully, each 15 liters of substratum (fermention medium composition: glycerine 2.0% of packing into, analysis for soybean powder 2.0%, NaCl0.3%, citrulline 0.01%, CoCl0.002%, pH7.0), behind 121 ℃ of 30 minutes steam sterilizings, in each fermentor tank, plant 6 bottles of above-mentioned seeds, 15 liters/minute of stirring velocity 250r.p.m air flows, 27 ℃ of cultivations in 120 hours.
Culture centrifugal (10,000rpm), separating thallus and nutrient solution, obtain 30 liters of supernatant liquors, with 6N HCl the pH value of above-mentioned clear liquid is transferred to 3.0, centrifugal again, obtain throw out, in throw out, add 15 liter of 50% acetone soln, stir, extract, filter, obtain extracting solution, it is evaporated to 6 liters, adds 3 liters of ethyl acetate, the vibration extraction secondary, the acetic acid ethyl acetate extract that obtains, concentrating under reduced pressure obtains the oily crude extract, adds the 750ml hexane in crude extract, extract and obtain hexane extraction liquid twice, concentrating under reduced pressure obtains raw product 4 gram, raw product silicagel column (mark's corporate system of packing into, ART7734,160 grams) chromatography, with hexane-ethyl acetate solution (30: 1) stripping, portioning is collected, each partly carries out bioassay with pine wilt nematode, active component merges, and concentrating under reduced pressure obtains 30 milligrams of raw product, be dissolved in the minimum of chloroform, get 1/20 amount sample introduction, (the mountain village chemical research is made, AM324(ODS with reverse separator column at every turn, 10 * 300mm) carry out high pressure liquid chromatography (HPLC), wash out with 60% acetonitrile, 2 milliliters of flow velocity per minutes collect to show the active component at retention time 18.5 minutes and each peak of 19.6 minutes, the concentrating under reduced pressure drying obtains guarding against platform mycin A7.2 milligram respectively and guards against platform mycin B5 milligram.
Description of drawings:
Fig. 1, the ultra-violet absorption spectrum (in hexanaphthene, measuring) of ring platform mycin A
Fig. 2, the infrared absorption spectrum (in KBr solution, measuring) of ring platform mycin A
Fig. 3, the nuclear magnetic resonance spectrum (in heavy chloroform, measuring) of the proton of ring platform mycin A
Claims (1)
1, a kind of antiparasitic new antibiotic with following structural I is guarded against the manufacture method of platform mycin, it is characterized in that cultivating streptomycete KP-197 bacterial strain (streptomyces spkp-197, CGMCC NO.0130), isolate then and guard against the platform mycin, the structural formula I:
(R=sec.-propyl in the formula)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 88100187 CN1027905C (en) | 1988-01-27 | 1988-01-27 | Preparation method of antiparasitic new antibiotic and its salts |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 88100187 CN1027905C (en) | 1988-01-27 | 1988-01-27 | Preparation method of antiparasitic new antibiotic and its salts |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1034580A CN1034580A (en) | 1989-08-09 |
| CN1027905C true CN1027905C (en) | 1995-03-15 |
Family
ID=4831221
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 88100187 Expired - Fee Related CN1027905C (en) | 1988-01-27 | 1988-01-27 | Preparation method of antiparasitic new antibiotic and its salts |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1027905C (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102533869B (en) * | 2012-01-13 | 2014-12-17 | 高丙利 | Actinomycete preparation and application thereof in aspect of controlling plant parasitic nematodes |
| CN107501131A (en) * | 2017-08-30 | 2017-12-22 | 山东省农药科学研究院 | One kind guards against platform mycin analog and preparation method thereof |
-
1988
- 1988-01-27 CN CN 88100187 patent/CN1027905C/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN1034580A (en) | 1989-08-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| DK165436B (en) | ANIMAL FEEDING | |
| JPH0511956B2 (en) | ||
| FI94262B (en) | Process for preparing macrolide against parasites | |
| CN1221752A (en) | Novel lantibiotic related to actagardine, and process for preparation and use thereof | |
| CN1027905C (en) | Preparation method of antiparasitic new antibiotic and its salts | |
| US4229535A (en) | Method for preparing multhiomycin | |
| DK170282B1 (en) | Antibiotics LL-E19020-alpha and LL-E19020-beta, process for their preparation, biologically pure Streptomyces lydicus culture, process for increasing the growth rate of meat-producing animals and fish, increasing feed utilization efficiency and / or improving milk yield in lactating ruminants. and composition for treating or preventing protozoan infections | |
| CN87100250A (en) | Preparation method of new antibiotic aureonucleomycin and agricultural bactericide | |
| CA1183789A (en) | Antibiotic 76-11, process for the production thereof, anticoccidiosis agent and domestic animals growth accelerator comprising the same as an effective ingredient | |
| CN1009278B (en) | Acid polycyclic ether antibiotic with the function of anticoccidiosis and growth promotion | |
| CN1045997C (en) | Microbiological process for making UK-61,689 | |
| US5073369A (en) | Efomycins as performance promoters in animals | |
| US4670260A (en) | Antibiotic for animal feeds | |
| EP0053025B1 (en) | Antibiotic and its preparation from a streptomyces microorganism | |
| CN1028436C (en) | Process for preparing antibiotic "Zhijiangjunsu" and its medicine for preventing and curing diseases of animals and fowls | |
| JPH0784472B2 (en) | Antibiotic A80190 | |
| SE437917B (en) | ANIMAL FEEDS INCLUDING A NEW CHEMICAL SOCIETY THAT PROMOTES GROWTH AND IMPROVES ANIMAL FEEDING APPROPRIATE | |
| CN1059678C (en) | Substance IT-62-B | |
| CA2092402C (en) | Anticoccidial and growth promoting polycyclic ether antibiotic | |
| CN1022765C (en) | Preparation method of cell variety mycomycin of new antibiotic | |
| US5155097A (en) | Polycyclic ether antibiotic having anthelmintic, anticoccidial and growth promotant activity | |
| CN1404484A (en) | Pluraflavins and derivatives thereof, process for their preparation and use thereof | |
| EP0158179A2 (en) | Antibiotic 6270, process for its production, and its use as an anticoccidiosis agent and a feed additive | |
| CN1878788A (en) | Substances K01-0509 and process for producing the same | |
| CN1014527B (en) | Prepn method of new antibiotic sr-1223 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C06 | Publication | ||
| PB01 | Publication | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |