CS272206B2 - Method of alpha-ergocryptine production - Google Patents
Method of alpha-ergocryptine production Download PDFInfo
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- CS272206B2 CS272206B2 CS853919A CS391985A CS272206B2 CS 272206 B2 CS272206 B2 CS 272206B2 CS 853919 A CS853919 A CS 853919A CS 391985 A CS391985 A CS 391985A CS 272206 B2 CS272206 B2 CS 272206B2
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- ergocryptine
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- YDOTUXAWKBPQJW-UHFFFAOYSA-N alpha-Ergocryptinine Natural products C1=CC(C=2C(N(C)CC(C=2)C(=O)NC2(C(=O)N3C(C(N4CCCC4C3(O)O2)=O)CC(C)C)C(C)C)C2)=C3C2=CNC3=C1 YDOTUXAWKBPQJW-UHFFFAOYSA-N 0.000 title claims description 8
- 229950001817 alpha-ergocryptine Drugs 0.000 title claims description 8
- YDOTUXAWKBPQJW-NSLWYYNWSA-N alpha-ergocryptine Chemical compound C1=CC(C=2[C@H](N(C)C[C@@H](C=2)C(=O)N[C@]2(C(=O)N3[C@H](C(N4CCC[C@H]4[C@]3(O)O2)=O)CC(C)C)C(C)C)C2)=C3C2=CNC3=C1 YDOTUXAWKBPQJW-NSLWYYNWSA-N 0.000 title claims description 8
- 238000000034 method Methods 0.000 title claims description 5
- 238000004519 manufacturing process Methods 0.000 title description 4
- 238000000855 fermentation Methods 0.000 claims description 10
- 230000004151 fermentation Effects 0.000 claims description 10
- 244000005700 microbiome Species 0.000 claims description 4
- 229910052500 inorganic mineral Chemical class 0.000 claims description 3
- 239000011707 mineral Chemical class 0.000 claims description 3
- 241000124224 Claviceps paspali Species 0.000 claims description 2
- 150000001720 carbohydrates Chemical class 0.000 claims description 2
- 235000014633 carbohydrates Nutrition 0.000 claims description 2
- 239000003630 growth substance Chemical class 0.000 claims description 2
- 229910017464 nitrogen compound Inorganic materials 0.000 claims description 2
- 150000002830 nitrogen compounds Chemical class 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 150000003839 salts Chemical class 0.000 claims description 2
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 9
- 239000002609 medium Substances 0.000 description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 5
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 5
- 229930006000 Sucrose Natural products 0.000 description 5
- 229910052799 carbon Inorganic materials 0.000 description 5
- 239000005720 sucrose Substances 0.000 description 5
- 229920001817 Agar Polymers 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 4
- 235000011130 ammonium sulphate Nutrition 0.000 description 4
- 229940041514 candida albicans extract Drugs 0.000 description 4
- 239000012138 yeast extract Substances 0.000 description 4
- ZHJGWYRLJUCMRT-UHFFFAOYSA-N 5-[6-[(4-methylpiperazin-1-yl)methyl]benzimidazol-1-yl]-3-[1-[2-(trifluoromethyl)phenyl]ethoxy]thiophene-2-carboxamide Chemical compound C=1C=CC=C(C(F)(F)F)C=1C(C)OC(=C(S1)C(N)=O)C=C1N(C1=C2)C=NC1=CC=C2CN1CCN(C)CC1 ZHJGWYRLJUCMRT-UHFFFAOYSA-N 0.000 description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 239000011591 potassium Substances 0.000 description 3
- 229910052700 potassium Inorganic materials 0.000 description 3
- RZLVQBNCHSJZPX-UHFFFAOYSA-L zinc sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Zn+2].[O-]S([O-])(=O)=O RZLVQBNCHSJZPX-UHFFFAOYSA-L 0.000 description 3
- QPFMBZIOSGYJDE-UHFFFAOYSA-N 1,1,2,2-tetrachloroethane Chemical compound ClC(Cl)C(Cl)Cl QPFMBZIOSGYJDE-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 241000221760 Claviceps Species 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- NHJPVZLSLOHJDM-UHFFFAOYSA-N azane;butanedioic acid Chemical compound [NH4+].[NH4+].[O-]C(=O)CCC([O-])=O NHJPVZLSLOHJDM-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 2
- 235000010755 mineral Nutrition 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- XJOOMMHNYOJWCZ-UHFFFAOYSA-N Agroclavine Natural products C1=CC(C2C=C(C)CN(C2C2)C)=C3C2=CNC3=C1 XJOOMMHNYOJWCZ-UHFFFAOYSA-N 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 description 1
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- NTIZESTWPVYFNL-UHFFFAOYSA-N Methyl isobutyl ketone Chemical compound CC(C)CC(C)=O NTIZESTWPVYFNL-UHFFFAOYSA-N 0.000 description 1
- UIHCLUNTQKBZGK-UHFFFAOYSA-N Methyl isobutyl ketone Natural products CCC(C)C(C)=O UIHCLUNTQKBZGK-UHFFFAOYSA-N 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241001149258 Sporobolus alterniflorus Species 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 229960003133 ergot alkaloid Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000004362 fungal culture Methods 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 150000004688 heptahydrates Chemical class 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 229930015720 peptide alkaloid Natural products 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 1
- 210000003934 vacuole Anatomy 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- -1 zinc cations Chemical class 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/18—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
- C12P17/182—Heterocyclic compounds containing nitrogen atoms as the only ring heteroatoms in the condensed system
- C12P17/183—Heterocyclic compounds containing nitrogen atoms as the only ring heteroatoms in the condensed system containing an indolo[4,3-F,G]quinoleine nucleus, e.g. compound containing the lysergic acid nucleus as well as the dimeric ergot nucleus
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- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Description
Vynález 9e týká způsobu výroby alfa-ergokryptinu kultivaci určitého druhu mikroorganismu· alfa-Ergokryptin » 9,10alfa-dihydro-12'~ hydroxy-2í/l-n>ethylethyl/-5'alfa-/2-methylpropyl/-ergotaman-3', 6', 18-trion je, jak známo, farmakologicky účinný. Podle dosud známých způsobů výroby je alfa-ergokryptin dostupný fermenťační cestou jenom ve směsi s ostatními námelovými a peptidovýmí alkaloidy /německý zveřejňovaci spis 31 04 151/. Tim je jeho výrobě iizolace velice náročnáThe invention 9e relates to a method for producing alpha-ergocryptine by culturing a particular kind of microorganism. Alpha-Ergocryptine 9,10 alpha- dihydro-12'-hydroxy-2H-ethylethyl-5'alpha- (2-methylpropyl) -ergotaman-3 ' The 6 ', 18-trione is known to be pharmacologically active. According to the known production methods, alpha-ergocryptine is available via fermentation only in admixture with other ergot and peptide alkaloids (German publication 31 04 151). Tim is very difficult to manufacture and insulate
Nyní bylo nalezeno, že kmen houby druhu Clavioeps vylučuje do kultivačního prostředí ve vysokých výtěžcích alfa-ergokryptin prostý vedle stop agroklavinu přítomnosti dalších námelových alkaloidů. Tento kmen druhu Claviceps byl izolován z námele divoce rostoucí traviny druhu Spartina alterniflora; má interní označeni Schering, MBCE 5227 a je uložen v německé sbírce pro mikroorganismy pod číslem OSM 2836. Kmen vytváří na agarových prostředcích se sacharózou jako zdrojem uhlíku a aeparagínea jako zdrojem dusíku ploché,· nažloutlé kolonie z kompaktního mycelia. Průměr kolonii činí po 7 dnech kultivace 2 až 3 cm a po 20 dnech kultivace 6 až 8 cm.It has now been found that a fungus strain of the species Clavioeps secretes alpha-ergocryptin free from traces of agroclavin in the presence of other ergot alkaloids in high yields. This Claviceps strain was isolated from the ergot of wild-growing Spartina alterniflora; has the internal designation Schering, MBCE 5227 and is deposited in the German Collection for Microorganisms under number OSM 2836. The strain forms flat, yellowish colonies of compact mycelium on agar compositions with sucrose as a carbon source and aeparagine as a nitrogen source. The colony diameter is 2-3 cm after 7 days of cultivation and 6-8 cm after 20 days of cultivation.
Na agarovém'živičném prostředí se sacharózou jako zdrojem uhlíku a jantaranera amonným nebo citranem amonným jako zdrojem dusíku vytváří houba pomalu rostouc!,· klenuté a nad agarovým povrchem lehce nadzdvižené, nažloutle hnědé kolonie. Kolonie má tvorbou vzdušného mycelia hrubý,· kulovitý povrch. Okraj kolonie je roztřepen. Průměr kolonie čini po 14 dnech 2 až 3 cm a po době 30 dní 6 až 7 cm. Vedle hyfovitého mycelia se vyskytuji krátké kulovité buňky,· které mohou být též nepravidelně tvarované a často vytvářejí řetězy a vyplněné vakuolami silně lámou světlo. Průměr hyf čini 3 až 4 um. Vakuolizované buňky mají průměr od 6 do 10 £um a délku od 10 do 25 <um.In an agar medium with sucrose as the carbon source and ammonium succinate or ammonium citrate as the nitrogen source, the fungus forms a slowly growing, domed and slightly elevated, yellowish brown colony above the agar surface. The colony has a rough, spherical surface formation of the airy mycelium. The colony's edge is frayed. The colony diameter is 2-3 cm after 14 days and 6-7 cm after 30 days. In addition to the hyphal mycelium, there are short spherical cells, which may also be irregularly shaped and often form chains and filled with vacuoles strongly refract the light. The diameter of the hyphae is 3 to 4 µm. Vacuolated cells have a diameter of 6 to 10 microns and a length of £ 10 to 25 <m.
Způsob výroby alfa-ergokryptinu podle vynálezu spočívá v tom,že se mikroorganismus druhu Claviceps paspali OSM 2836 kultivuje za anerobních podmínek v přítomnosti uhlohydrátů, dusíkatých sloučenin, růstových látek β minerálních soli při hodnotě pH 4 až 6 za teploty 20 až 35 °C a po skončené fermentaci se vzniklý alfa-ergokryptin izoluje.The process for the preparation of alpha-ergocryptin according to the invention is characterized in that the microorganism of the species Claviceps paspali OSM 2836 is cultivated under anerobic conditions in the presence of carbohydrates, nitrogen compounds, β mineral salt growth substances at pH 4-6 at 20-35 ° C. after fermentation, the resulting alpha-ergocryptin is isolated.
Způsob podle vynálezu se tedy provádí za podmínek, kterých se obvykle používá k pěstováni kultur hub pro metsbolickou syntézu. Tak se nejprve urči v obecných obvyklých předběžných pokusech nejpřiznivějši podminky fermentace, jako například volba nejpřlznivějšího živného prostředí/ technických podmínek/ jako teploty, vzdušnění, přesné hodnoty pH a optimálních časů pro klíčeni a vývoj mikroorganismu.Thus, the process of the invention is carried out under the conditions commonly used to cultivate fungal cultures for metsbolic synthesis. Thus, the most favorable fermentation conditions, such as selection of the most favorable nutrient medium / technical conditions / such as temperature, aeration, exact pH and optimum germination and development times, are first determined in general conventional preliminary experiments.
□ako zdroj uhlíku pro fermentační prostředí lze použit například glukózu nebo sacharozu. Oako zdroj dusíku slouží kromě jiného asparagin, jantaran amonný nebo síran amonný. Prostředí obsahuje dále nezbytné růstové látky /například kvasničný extrakt/ a minerální látky /draselné, hořečnaté, vápenaté, železnaté a zinečnaté kationty, jakož i sulfátové, fosfátové, dusičnanové g chloridové aionty/ v obvykle používané koncentraci.Například for example, glucose or sucrose can be used as a carbon source for the fermentation medium. Oako's nitrogen source serves inter alia asparagine, ammonium succinate or ammonium sulfate. The medium further contains the necessary growth agents (e.g., yeast extract) and minerals (potassium, magnesium, calcium, ferrous and zinc cations, as well as sulfate, phosphate, nitrate, chloride and ionic ions) in the concentration normally used.
Fermentace může být jedno- nebo dvouetupňové, přičemž prostředí užité pro předkulturu může být totožné e prostředím hlavní kultury nebo odliěné. Pro předkulturu se používá 8 výhodou glukóza jako zdroj uhlíku, pro hlavni kulturu s výhodou saoharóza. Prostředí předkultury obsahuje s výhodou 10 až 100 mg/1 zdrojů uhlíku/ prostředí hlavní kultury s výhodou 100 až 300 mg.The fermentation can be one- or two-stage, with the medium used for the preculture being identical to that of the main culture or differentiated. For preculture, preferably glucose is used as a carbon source, for the main culture preferably saoharose. The preculture environment preferably contains 10 to 100 mg / l of carbon source / main culture medium, preferably 100 to 300 mg.
Na začátku fermentace se nastavuje hodnota pH prostředí s výhodou v rozsahu od 4 do So Teplota kultivace ss pohybuje v rozmezí asi od 10 až do 35 °C, s výhodou v rozmezí od 20 do 30 °C. Podminky kultivace jsou přísně aerobní. Optimální doba fermentace se zjišťuje obvyklým způsobem pomoci analýzy vzniklého alfa-ergokryptinu.At the beginning of the fermentation is adjusted pH of the medium preferably ranges from 4 to about S ss cultivation temperature ranges from about 10 to 35 ° C, preferably from 20 to 30 ° C. The cultivation conditions are strictly aerobic. The optimal fermentation time is determined in a conventional manner by analysis of the resulting alpha-ergocryptine.
Po skončené fermentaci ee vzniklý alfaergokryptin izoluje o sobě známým způsobem, například tak,'· že se fermentační násady extrahuji s vodou nemisitelným organickým rozpouštědlem,· jako ethylacetátem, methylisobutylketonem, dichlormethanem, chloroformem nebo tetrachlorethanem,- extrakty se zkoncsntruji a získaný surový produkt se čisti chromatografií a/nebo krystalizaci.After fermentation, the resulting alphaergocryptine is isolated in a manner known per se, for example by extracting the fermentation broth with a water-immiscible organic solvent, such as ethyl acetate, methyl isobutyl ketone, dichloromethane, chloroform or tetrachloroethane, and extracting the crude product obtained. chromatography and / or crystallization.
Následující příklad provedeni slouží k vysvětleni způsobu podle vynálezu.The following exemplary embodiment serves to explain the method of the invention.
CS 272208 B2CS 272208 B2
Claviceps druh OSM 2836 aa pěstuje na živném prostředí» které obsahuje následující složky:Claviceps species OSM 2836 aa is grown on a nutrient medium »which contains the following ingredients:
Sacharózu /100g/l/,kyselinu citrónovou /10g/l/, kvasničný extrakt /0,!l g/1/,1 dihydroganfosfát draselný /500 mg/l/,< heptahydrát síranu hořečnatého /300 mg/1/,’ síran amonný /6 g/1/, tetrahydrót dusičnanu vápenatého /1 g/1/, heptahydrát síranu železnatého /7 mg/1/, heptahydrát síranu zinečnatáho /6 mg/1/, agar /16 g/l/.Hodnota pH živného prostředí se nastavuje na 5,1. Pěstovaná kultura ea uchovává po dobu 5 až 20 dnů při teplotě 30 °C v ter» mostatu.Sucrose / 100 g / l /, citric acid / 10 g / l /, yeast extract / 0 ug / 1/1 dihydroganfosfát potassium / 500 mg / l / <heptahydrate of magnesium sulfate / 300 mg / 1 /, 'ammonium sulfate (6 g / l), calcium nitrate tetrahydrate (1 g / l), ferrous sulfate heptahydrate (7 mg / l), zinc sulfate heptahydrate (6 mg / l), agar (16 g / l). set to 5.1. The cultivated culture ea is stored for 5 to 20 days at 30 ° C in teratate.
Asi 1 om2 velký kousek mycelia se za sterilních podmínek rozmělni pomoci mixéru v 5 ml fyziologického roztoku a tim se očkuje 50 ml předkultury obsahující glukózu /100 g/1/, kyselinu citrónovou /7,5 g/1/, dihydrohenfosfát draselný /0,59 g/1/,' heptahydrát síranu hořečnatého /300 mg/1/, síran amonný /6 g/1/, heptahydrát síranu železnatého /7 mg/1/, heptahydrát síranu zinečnatého /7 mg/1// tetrahydřát dusičnanu vápenatého /1 g/1/, kvasničný extrakt /0,1 g/1/, nastavené na hodnotě pH 5,1, která je umístěna v·’.500 ml Erlenmeysrově baňce a kultivuje se na rotační třepačce s 220 otáčkami za minutu po dobu 4 dní při teplotě 24 °C.About 1 om 2 large piece of mycelium under sterile conditions comminuted means of a mixer in 5 ml of physiological solution and thus is inoculated with 50 ml of a preculture containing glucose / 100 g / 1 /, citric acid / 7.5 g / 1 /, potassium dihydrohenfosfát / 0 59 g (1), magnesium sulfate heptahydrate (300 mg / 1), ammonium sulfate (6 g / 1), ferrous sulfate heptahydrate (7 mg / 1), zinc sulfate heptahydrate (7 mg / 1 // calcium nitrate tetrahydrate) (1 g / l), yeast extract (0.1 g / l), adjusted to pH 5.1, placed in a 500 ml Erlenmeyser flask and cultured on a rotary shaker at 220 rpm for a period of time. 4 days at 24 ° C.
ml takto získané předkultury se přenesou do 80 ml prostředí obsahujícího sacharózu /200 g/l/r kyselinu citrónovou /10 g/1/, kvasničný extrakt /0,1 g/1/, dihydrogenfosfát draselný /500 mg/1/, heptahydrát síranu hořečnatého /300 mg/1/, síran amonný /6 g/1/, tetrahydřát dusičnanu vápenatého /1 g/1/, heptahydrát síranu železnatého /7 mg/1/ a heptahydrátu síranu zinečnatáho /8 mg/1/, nastaveného na hodnotu pH 5,1,' které Js umístěno v 500 ml Erlenmeyerově baňce a po dobu 9 dni se při teplotě 24 °C třepě na rotační třepačce se 240 otáčkami za minutu.ml of the preculture thus obtained are transferred to 80 ml of sucrose-containing medium (200 g / l) of citric acid (10 g / l), yeast extract (0.1 g / l), potassium dihydrogen phosphate (500 mg / l), sulfate heptahydrate magnesium (300 mg / l), ammonium sulfate (6 g / l), calcium nitrate tetrahydrate (1 g / l), ferrous sulfate heptahydrate (7 mg / l) and zinc sulfate heptahydrate (8 mg / l), set to pH 5.1, placed in a 500 ml Erlenmeyer flask and shaken for 9 days at 24 ° C on a rotary shaker at 240 rpm.
Potom se živné prostředí odfiltruje a pomocí vysokotlaké kapalinové chromatografie /Fresenius Z,Anal.Chem, 303,' 1980, 208/ ss urči obsah alfsargokryptinu. Koncentrace kultivačního filtrátu čini 525 mg/1.The culture medium is then filtered off and the alfsargocryptin content is determined by high pressure liquid chromatography (Fresenius Z, Anal.Chem, 303, 1980, 208). The concentration of the culture filtrate was 525 mg / L.
Claims (1)
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19843420953 DE3420953A1 (en) | 1984-06-01 | 1984-06-01 | METHOD FOR PRODUCING (ALPHA) ERGOKRYPTIN |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CS391985A2 CS391985A2 (en) | 1990-04-11 |
| CS272206B2 true CS272206B2 (en) | 1991-01-15 |
Family
ID=6237689
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CS853919A CS272206B2 (en) | 1984-06-01 | 1985-05-31 | Method of alpha-ergocryptine production |
Country Status (7)
| Country | Link |
|---|---|
| EP (1) | EP0183739B1 (en) |
| JP (1) | JPS61502374A (en) |
| CS (1) | CS272206B2 (en) |
| DD (1) | DD237677A5 (en) |
| DE (2) | DE3420953A1 (en) |
| HU (1) | HU198100B (en) |
| WO (1) | WO1985005635A1 (en) |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1133916A (en) * | 1966-07-22 | 1968-11-20 | Farmaceutica Italia Soc | Ergocryptine |
| SE332403B (en) * | 1966-07-22 | 1971-02-08 | Farm Italia Soc | |
| HU178405B (en) | 1980-02-08 | 1982-04-28 | Richter Gedeon Vegyeszet | Process for preparing ergocornine and beta-ergocryptine by fermentation |
| DE3104215A1 (en) * | 1981-02-06 | 1982-08-19 | Richter Gedeon Vegyészeti Gyár R.T., 1103 Budapest | Process for the preparation of alkaloids in controlled amount and controlled ratio of amounts by fermentation |
-
1984
- 1984-06-01 DE DE19843420953 patent/DE3420953A1/en not_active Withdrawn
-
1985
- 1985-05-30 WO PCT/DE1985/000193 patent/WO1985005635A1/en active IP Right Grant
- 1985-05-30 DD DD85276810A patent/DD237677A5/en not_active IP Right Cessation
- 1985-05-30 DE DE8585902482T patent/DE3567226D1/en not_active Expired
- 1985-05-30 HU HU852884A patent/HU198100B/en not_active IP Right Cessation
- 1985-05-30 EP EP85902482A patent/EP0183739B1/en not_active Expired
- 1985-05-30 JP JP60502496A patent/JPS61502374A/en active Pending
- 1985-05-31 CS CS853919A patent/CS272206B2/en not_active IP Right Cessation
Also Published As
| Publication number | Publication date |
|---|---|
| EP0183739A1 (en) | 1986-06-11 |
| DE3420953A1 (en) | 1985-12-05 |
| CS391985A2 (en) | 1990-04-11 |
| WO1985005635A1 (en) | 1985-12-19 |
| DD237677A5 (en) | 1986-07-23 |
| EP0183739B1 (en) | 1989-01-04 |
| HU198100B (en) | 1989-07-28 |
| JPS61502374A (en) | 1986-10-23 |
| DE3567226D1 (en) | 1989-02-09 |
| HUT38396A (en) | 1986-05-28 |
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| Date | Code | Title | Description |
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| MK4A | Patent expired |
Effective date: 20000531 |