WO1992011231A1 - Phenylpolyenamides, process for producing the same and agents - Google Patents

Phenylpolyenamides, process for producing the same and agents Download PDF

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Publication number
WO1992011231A1
WO1992011231A1 PCT/EP1991/002480 EP9102480W WO9211231A1 WO 1992011231 A1 WO1992011231 A1 WO 1992011231A1 EP 9102480 W EP9102480 W EP 9102480W WO 9211231 A1 WO9211231 A1 WO 9211231A1
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Prior art keywords
fenalamide
aromatic
methanol
water
protons
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PCT/EP1991/002480
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German (de)
French (fr)
Inventor
Gerhard Höfle
Norbert Bedorf
Rolf Jansen
Heinrich Steinmetz
Wolfram Trowitzsch-Kienast
Hans Reichenbach
Edgar Forche
Klaus Gerth
Herbert Irschik
Brigitte Kunze
Florenz Sasse
Gerhard Hunsmann
Elke Jurkiewicz
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GESELLSCHAFT FüR BIOTECHNOLOGISCHE FORSCHUNG MBH (GBF)
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Publication of WO1992011231A1 publication Critical patent/WO1992011231A1/en

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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N49/00—Biocides, pest repellants or attractants, or plant growth regulators, containing compounds containing the group, wherein m+n>=1, both X together may also mean —Y— or a direct carbon-to-carbon bond, and the carbon atoms marked with an asterisk are not part of any ring system other than that which may be formed by the atoms X, the carbon atoms in square brackets being part of any acyclic or cyclic structure, or the group, wherein A means a carbon atom or Y, n>=0, and not more than one of these carbon atoms being a member of the same ring system, e.g. juvenile insect hormones or mimics thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C235/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
    • C07C235/02—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton
    • C07C235/32—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton containing six-membered aromatic rings
    • C07C235/34—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to acyclic carbon atoms and singly-bound oxygen atoms bound to the same carbon skeleton the carbon skeleton containing six-membered aromatic rings having the nitrogen atoms of the carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D303/00—Compounds containing three-membered rings having one oxygen atom as the only ring hetero atom
    • C07D303/02—Compounds containing oxirane rings
    • C07D303/38—Compounds containing oxirane rings with hydrocarbon radicals, substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D303/46—Compounds containing oxirane rings with hydrocarbon radicals, substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals by amide or nitrile radicals
    • 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
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/02—Amides, e.g. chloramphenicol or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes

Definitions

  • the invention relates to fenalamides of the following general formula:
  • the invention further relates to fenalamides of the following general formula:
  • the invention further relates to a fenalamide (A) which is characterized by one or more of the following parameters:
  • the invention further relates to a process for the preparation of fenalamides, in particular the fenalamides A, B and / or C mentioned above, in which - Myxococcus stipitatus DSM 6259 cultivated in a medium containing carbon sources, nitrogen sources and mineral salts in the presence of an adsorber resin,
  • the adsorber resin is separated off and extracted with methanol
  • Fenalamide A, B and / or C can be treated with UV light to prepare cis-trans isomers of fenalamides A, B and / or C or of fenalamides of one or more of the general formulas. subject and separate the isomers obtained chromatographically and isolate if necessary.
  • the invention further relates to a therapeutic agent (except for combating viral diseases) consisting of at least one fenalamide or containing at least one
  • Fenalamide if necessary in addition to a conventional carrier and / or diluent.
  • This agent can be used to combat fungal or parasitic diseases of canteens and animals.
  • the invention relates to an agent for crop protection for agriculture, forestry and / or horticulture,
  • At least one fenalamide optionally in addition to a conventional carrier and / or diluent.
  • the bacterium Myxococcus stipitatus belongs to the order of the
  • Myxococcaceae The production strain Myxococcus stipitatus Mx s40 was obtained at GBF in September 1988 from a soil sample from Jacobsburg State Park near Wind Gap, Pennsylvania. USA, isolated, put down at the German Collection of Microorganisms (DSM) under the number DSM 6259.
  • DSM German Collection of Microorganisms
  • the stock culture is carried out on agar plates, preferably on yeast agar (VY / 2 agar).
  • This medium contains 0.5% baker's yeast, 0.1% CaCl2 ⁇ H2O, 0.1 ⁇ g / l vitamin B12 and 1.2% agar.
  • the pH is adjusted to 7.2.
  • the plates are at 30 ; incubated.
  • the vegetative sticks are slender and have slightly tapered ends. They are about 0.7 ⁇ m thick and 2 to 5 ⁇ m long.
  • the Myxospores are oval, highly refractive, about 1.1 to 1.3 ⁇ 1.3 to 1.5 ⁇ m in size. Due to the sliding movement of the
  • Irradiation with ultraviolet light with a wavelength of 366 nm shows a swarm colony a few days old showing yellow fluorescence.
  • the Mx s40 strain produces substances, namely fenalamides, that inhibit the growth of certain fungi and bacteria (cf.
  • the inhibitors can be isolated both from the cells and from the culture supernatant.
  • the substances are produced during the logarithmic to the stationary phase.
  • a fermentor with 700 1 working volume is filled with 630 1 culture medium (composition: 0.6% peptone from casein, (tryptically digested) 0.05% CaCl2 ⁇ 2H2O, 0.2% MgSO4 ⁇ 7H2O, pH 7.2 adjusted with H2 SO4).
  • composition 0.6% peptone from casein, (tryptically digested) 0.05% CaCl2 ⁇ 2H2O, 0.2% MgSO4 ⁇ 7H2O, pH 7.2 adjusted with H2 SO4
  • Released inhibitors are the medium 1 to 2% (V / V) of an adsorbent resin e.g. XAD-1180 added. Inoculated with 70 1 one in the same medium in a correspondingly smaller one
  • Fermentor produced a day-old preculture.
  • the fermentation takes place at 30 ° with a stirring speed of 150 rpm and aeration of 0.2 V / Vm.
  • Initial foam formation is prevented by adding 70ml of silicone anti-foam, for example tegosipon.
  • the pH is kept at 7.2 by controlled addition of 5% H2SO4.
  • the fermentation is ended after three days.
  • the fermentation is carried out in the presence of 81 Asorber resin XAD-1180. Towards the end of the fermentation, the resin is separated from the medium and the cells through a sieve. The resin is eluted in a column with 3 bed volumes of methanol. After concentrating on a rotary evaporator, 234 g of crude extract are obtained. The crude extract is dissolved in 20 l of ethyl acetate and 20 l of water and distributed in a countercurrent centrifugation. The water phase is discarded, the ethyl acetate phase is dried and concentrated in a water jet vacuum. Yield ethyl acetate phase: 38 g.
  • UV (methanol): ⁇ max (1g ⁇ ) 356 nm (4.77)
  • Fenalamides inhibit the growth of fungi and bacteria, mainly gram-positive bacteria (see table)

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  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Pest Control & Pesticides (AREA)
  • Environmental Sciences (AREA)
  • Insects & Arthropods (AREA)
  • Agronomy & Crop Science (AREA)
  • Plant Pathology (AREA)
  • Dentistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Peptides Or Proteins (AREA)

Abstract

Phenylpolyenamides having general formula (I), a process for producing the same and agents containing phenylpolyenamides are disclosed. The dotted line between the C10 and the C11 atoms may be a (further) C-C bond or an -O- bridge and the C8 atom may carry a methyl group.

Description

PHENYLPOLYENAMIDE, HERSTELLUNGSVERFAHREN UND MITTEL  PHENYL POLYEN AMIDES, PRODUCTION METHOD AND MEDIUM
Die Erfindung" betrifft Fenalamide der folgenden allgemeinen Formel: The invention relates to fenalamides of the following general formula:
Figure imgf000003_0001
Figure imgf000003_0001
wobei die gestrichelte Linie zwischen dem C10- und dem C11-Atom eine (weitere) C-C-Bindung oder eine -O-Brücke sein kann und wobei aas C8-Atom eine Methylgruppe tragen kann. Ferner betrifft die Erfindung Fenalamide der folgenden allgemeinen Formel:wherein the dashed line between the C10 and the C11 atom can be a (further) CC bond or an -O bridge and wherein the C8 atom can carry a methyl group. The invention further relates to fenalamides of the following general formula:
Figure imgf000004_0001
Ferner betrifft die Erfindung ein Fenalamid (A), das durch einen oder mehrere der folgenden Parameter gekennzeichnet ist:
Figure imgf000004_0001
The invention further relates to a fenalamide (A) which is characterized by one or more of the following parameters:
HPLC (Säule: Nucleosil-C18, 5μ, 12.5 cm × 4 mm; Laufmittel: Methanol:Wasser= 85/15; Durchfluß: 1.0 ml/ min; Detektion: Diodenarray-System HP 1040A): Retentionszeit Rt= 6.5 min. - UV (Methanol.: λmax (1g ε) = 356 nm (4.77) HPLC (column: Nucleosil-C 18 , 5μ, 12.5 cm × 4 mm; mobile solvent: methanol: water = 85/15; flow rate: 1.0 ml / min; detection: diode array system HP 1040A): retention time Rt = 6.5 min. - UV (methanol .: λ max (1g ε) = 356 nm (4.77)
[α]D 25= - 189° (c = 1, in Methanol) [α] D 25 = - 189 ° (c = 1, in methanol)
1H-NMR (600 MHz, CD3OD): δ = 7.24 [t, J= 7.7 Hz, 2H, meta-Protonen am Aromaten], 7.16 [d, J = 7.2 Hz, 2H, ortho-Protonen am Aromaten], 7.14 [t, J = 7.3 Hz, 1H, para-Proton am Aromat], 7.07 [dd, J= 11.0 und 14.5 Hz, 1H, 5-H], 7.02 [d, J = 11.4 Hz, 1H, 3-H], 7.73 [dd, J = 10.6 und 15.1 Hz, 1H, 8-H], 6.59 [dd, J = 11.4 und 14.5 Hz, 1H, 4-H], 6.40 [d, J = 15.1 Hz, 1H, 9-H], 6.14 [m, 2H, 6- und 7-H (durch 2D-H-COSY-Spektren ermittelt), 5.63 [d, J = 9.7 Hz, 1H, 11-H], 5.27 [d, J = 9.6 Hz, 1H, 15-H], 4.09 [dq, J - 5.7 und 12.4 Hz, 1H, 1'-H], 3.85 [d, J = 6.7 Hz, 1H, 13-H], 5.72 [dq, J = 5.8 und 11.0 Hz, 2H, 2'-H2], 2.83 [m, 1H, 12-H], 2.64 [m, 1H, 18Ha], 2.52 [m, 1H, 18Hb] , 2.45 [m, 1H, 16-H], 2.02 [d, J = 0.8 Hz, 3H, 2-CH3], 1.89 [d, J = 1.0 Hz, 3H, 10-CH3], 1.68 [m, 1H, 17-HJ, 1.62 [d, J = 1.2 Hz, 3H, 14-CH3], 1.54 [m, 1H, 17-Hb], 1.23 [d, J = 6.8 Hz, 3H, 1'-CH3], 1.00 [d, J = 1.0 Hz, 3H, 16-CH3], 0.97 [d, JJ = 6.9 Hz, 3H, 12-CH3]. - 1 H-NMR (600 MHz, CD 3 OD): δ = 7.24 [t, J = 7.7 Hz, 2H, meta-protons on the aromatic], 7.16 [d, J = 7.2 Hz, 2H, ortho-protons on the aromatic] , 7.14 [t, J = 7.3 Hz, 1H, para-proton on aromatic], 7.07 [dd, J = 11.0 and 14.5 Hz, 1H, 5-H], 7.02 [d, J = 11.4 Hz, 1H, 3- H], 7.73 [dd, J = 10.6 and 15.1 Hz, 1H, 8-H], 6.59 [dd, J = 11.4 and 14.5 Hz, 1H, 4-H], 6.40 [d, J = 15.1 Hz, 1H, 9-H], 6.14 [m, 2H, 6- and 7-H (determined by 2D-H-COZY spectra), 5.63 [d, J = 9.7 Hz, 1H, 11-H], 5.27 [d, J = 9.6 Hz, 1H, 15-H], 4.09 [dq, J - 5.7 and 12.4 Hz, 1H, 1'-H], 3.85 [d, J = 6.7 Hz, 1H, 13-H], 5.72 [dq, J = 5.8 and 11.0 Hz, 2H, 2'-H 2 ], 2.83 [m, 1H, 12-H], 2.64 [m, 1H, 18H a ], 2.52 [m, 1H, 18H b ], 2.45 [m , 1H, 16-H], 2.02 [d, J = 0.8 Hz, 3H, 2-CH 3 ], 1.89 [d, J = 1.0 Hz, 3H, 10-CH 3 ], 1.68 [m, 1H, 17- HJ, 1.62 [d, J = 1.2 Hz, 3H, 14-CH 3 ], 1.54 [m, 1H, 17-H b ], 1.23 [d, J = 6.8 Hz, 3H, 1'-CH 3 ], 1.00 [d, J = 1.0 Hz, 3H, 16-CH 3 ], 0.97 [d, JJ = 6.9 Hz, 3H, 12-CH 3 ]. -
EI-MS: m/Z (%) = 491 (3) [M]+, 473 (6) [M-H2O]+, 455 (2) [M-2 X H2O ] , 289 ( 100 ) [C18H27NO2 ] , 271 ( 30 ) [m/z = 289-H2O] . - Hochauflösung: gefunden 491.3381, für C32H45NO3 ber. 491.3399 Ferner betrifft die Erfindung ein Fenalamid (B), das durch einen oder mehrere der folgenden Parameter gekennzeichnet ist; EI-MS: m / Z (%) = 491 (3) [M] + , 473 (6) [MH 2 O] + , 455 (2) [M-2 XH 2 O], 289 (100) [C 18 H 27 NO 2 ], 271 (30) [m / z = 289-H 2 O]. - High resolution: found 491.3381, for C 32 H 45 NO 3 calc. 491.3399 The invention further relates to a fenalamide (B) which is characterized by one or more of the following parameters;
HPLC (Säule: Nucleosil-Cis, 5μ, 12.5 cm × 4 mm: Laufmittel: Methanol:Wasser= 85/15; Durchfluß: 1.5 ml/ min; Detektion: HPLC (column: Nucleosil-Cis, 5μ, 12.5 cm × 4 mm: mobile solvent: methanol: water = 85/15; flow rate: 1.5 ml / min; detection:
Diodenarray-System HP 1040A): Retentionszeit Rt = 9.6 min . - Diode array system HP 1040A): retention time R t = 9.6 min. -
UV (Methanol. : λmax (1g ε) = 346 nm (4.70). - 1H-NMR (600 MHz, [D6]Aceton): δ = 7.24 [t, J = 7.6 Hz, 2H, meta-Protonen am Aromat], 7.16 [d, J = 6.9 Hz, 2H, ortho-Protonen am Aromat], 7.14 [t, J = 7.3 Hz, 1H, para-Proton am Aromat], 6.95 [d, J = 11.4 Hz, 1H, 3-H], 6.60 [dd, J = 11.4 und 14.6 Hz, 1H, 4-H], 6.55 [dd, J = 14.6 und 10.0 Hz, 1H, 5-H], 6.42 [dd, J = 15.3 und 10.0 Hz, 1H, 6-H], 6.41 [d, J = 15.3 Hz, 1H, 7-H], 6.02 [s, 1H, 9-H], 5.47 [d, J = 9.6 Hz, 1H, 11-H], 5.52 [d, J = 9.6 Hz, 1H, 15-H], 4.03 [m, 1H, 1'-H], 3.82 [d, J = 6.0 Hz, 1H, 13- H], 3.61 [m, 2H, 2'-H2], 2.74 [m, 1H, 12-H], 2.61 [ddd, J = 5.5, 10.5 und 14.0 Hz, 1H, 18-Ha], 2.50 [ddd, J = 6.5, 10.0 und 14.0 Hz, 1H, 18-Hb], 2.44 [m, 1H, 16-H], 2.04 [d, J = 1.0 Hz, 3H, 2- CH3], 1.96 [d, J = 1.1 Hz, 3H, 8-CH3], 1.83 [d, J = 1.0 Hz, 3H, 10-CH3], 1.62 [d, J = 1.1 Hz, 3H, 14-CH3], 1.60 [m, 1H, 17-HJ, 1.51 [m, 1H, 17-Hb], 1.16 [d, J = 6.8 Hz, 3H, 1'-CH3], 0.96 [d, J = 6.5 Hz, 3H, 16-CH3], 0.95 [d, 6.8 Hz, 3H, 12-CH3]. - EI-MS (Die Ionen im Molpeak-Bereich sind nur sehr schwach. Im Gesamtspektrum ist m/z = 91 Basis-Ion. Das Ion m/z 487 beträgt hiervon ca. 5 %, die angegebenen Prozentzahlen beziehen sich nur auf den genannten Massenbereich) m/z (%) = 505 (17) [M]+, 487 (100) [M-H2O]+, 469 (74) [M-2 × H2O], 454 (48) [M-2 X H2O-CH3]. - Hochauflösung: gef. 505.3552 für C33H47NO3 ber. 505.3556 Ferner betrifft die Erfindung ein Fenalamid (C), das durch einen oder mehrere der folgenden Parameter gekennzeichnet ist. UV (methanol.: Λ max (1g ε) = 346 nm (4.70). - 1H-NMR (600 MHz, [D 6 ] acetone): δ = 7.24 [t, J = 7.6 Hz, 2H, meta-protons on Aromat], 7.16 [d, J = 6.9 Hz, 2H, ortho-protons on the aromatic], 7.14 [t, J = 7.3 Hz, 1H, para-proton on the aromatic], 6.95 [d, J = 11.4 Hz, 1H, 3-H], 6.60 [dd, J = 11.4 and 14.6 Hz, 1H, 4-H], 6.55 [dd, J = 14.6 and 10.0 Hz, 1H, 5-H], 6.42 [dd, J = 15.3 and 10.0 Hz, 1H, 6-H], 6.41 [d, J = 15.3 Hz, 1H, 7-H], 6.02 [s, 1H, 9-H], 5.47 [d, J = 9.6 Hz, 1H, 11-H ], 5.52 [d, J = 9.6 Hz, 1H, 15-H], 4.03 [m, 1H, 1'-H], 3.82 [d, J = 6.0 Hz, 1H, 13-H], 3.61 [m, 2H, 2'-H 2 ], 2.74 [m, 1H, 12-H], 2.61 [ddd, J = 5.5, 10.5 and 14.0 Hz, 1H, 18-H a ], 2.50 [ddd, J = 6.5, 10.0 and 14.0 Hz, 1H, 18-H b ], 2.44 [m, 1H, 16-H], 2.04 [d, J = 1.0 Hz, 3H, 2- CH 3 ], 1.96 [d, J = 1.1 Hz, 3H , 8-CH 3 ], 1.83 [d, J = 1.0 Hz, 3H, 10-CH 3 ], 1.62 [d, J = 1.1 Hz, 3H, 14-CH 3 ], 1.60 [m, 1H, 17-HJ , 1.51 [m, 1H, 17-H b ], 1.16 [d, J = 6.8 Hz, 3H, 1'-CH 3 ], 0.96 [d, J = 6.5 Hz, 3H, 16-CH 3 ], 0.95 [ d, 6 .8 Hz, 3H, 12-CH 3]. - EI-MS (The ions in the Molpeak range are only very weak. In the overall spectrum, m / z = 91 base ions. The ion m / z 487 is about 5%, the percentages given relate only to the ones mentioned Mass range) m / z (%) = 505 (17) [M] + , 487 (100) [MH 2 O] + , 469 (74) [M-2 × H 2 O], 454 (48) [M- 2 XH 2 O-CH 3 ]. - High resolution: found 505.3552 for C 33 H 47 NO 3 calc. 505.3556 The invention further relates to a fenalamide (C) which is characterized by one or more of the following parameters.
HPLC (Säule: Nucleosil-C18, 5μ, 12.5 cm × 4 mm; Laufmittel: Methanol :Wasser= 85/15: Durchfluß: 1.5 ml/ min; Detektion: HPLC (column: Nucleosil-C 18 , 5μ, 12.5 cm × 4 mm; mobile solvent: methanol: water = 85/15: flow rate: 1.5 ml / min; detection:
Diodenarray-System HP 1040A): Retentionszeit Rt = 3.8 min . - Diode array system HP 1040A): retention time R t = 3.8 min. -
UV (Methanol) : λmax (1g ε) = 328 nm (4.63). -UV (methanol): λ max (1g ε) = 328 nm (4.63). -
1H-NMR (300 MHz, CD3OD) : δ = 7.28 [t, J = 7.1 Hz, 2H, meta-Protonen am Aromat], 7.20 [d, J = 6.9 Hz, 2H, ortho-Protonen am Aromat], 7.18 [t, J = 7.1 Hz, 1H, para-Proton am Aromat], 6.96 [d, J = 11.0 Hz, 1H, 3-H], 6.60 [m (höherer Ordnung, 2H, 4- und 5-H], 6.64 [m, höherer Ordnung, 3H, 6-, 7- und 8-H], 5.35 [d, J = 9.6 Hz, 1H, 15-H], 4.08 [m, 1H, 1'-H], 3.92 [d, J = 10.4 Hz, 1H, 13-H], 3.62 [d, J = 9.8 Hz, 1H, 11-H], 3.57 [dq, J = 5.6 und 11.6 Hz, 2H, 2'-H2], 2.65 [m, 3H, 12-, 18-Ha und 18 Hb] , 2.51 [m, 2H, 18 Hb und 16-H], 2.01 [s, 3H, 2-CH3], 1.69 [s, 3H, 14-CH3], 1.61 [m, 1H, 17-HJ, 1.42 [s, 3H, 10-CH3], 1.33 [m, 1H, 17-Hb], 1.23 [d, J = 7.2 Hz, 3H, 1'-CH3], 1.02 [d, J = 6.8 Hz, 3H, 16-CH3], 0.99 [d, J = 7.3 Hz, 3H, 12-CH3]. - 1 H-NMR (300 MHz, CD 3 OD): δ = 7.28 [t, J = 7.1 Hz, 2H, meta-protons on the aromatic], 7.20 [d, J = 6.9 Hz, 2H, ortho-protons on the aromatic] , 7.18 [t, J = 7.1 Hz, 1H, para-proton on the aromatic], 6.96 [d, J = 11.0 Hz, 1H, 3-H], 6.60 [m (higher order, 2H, 4- and 5-H ], 6.64 [m, higher order, 3H, 6-, 7- and 8-H], 5.35 [d, J = 9.6 Hz, 1H, 15-H], 4.08 [m, 1H, 1'-H], 3.92 [d, J = 10.4 Hz, 1H, 13-H], 3.62 [d, J = 9.8 Hz, 1H, 11-H], 3.57 [dq, J = 5.6 and 11.6 Hz, 2H, 2'-H 2 ], 2.65 [m, 3H, 12-, 18-H a and 18 H b ], 2.51 [m, 2H, 18 H b and 16-H], 2.01 [s, 3H, 2-CH 3 ], 1.69 [ s, 3H, 14-CH 3 ], 1.61 [m, 1H, 17-HJ, 1.42 [s, 3H, 10-CH 3 ], 1.33 [m, 1H, 17-H b ], 1.23 [d, J = 7.2 Hz, 3H, 1'-CH 3 ], 1.02 [d, J = 6.8 Hz, 3H, 16-CH 3 ], 0.99 [d, J = 7.3 Hz, 3H, 12-CH 3 ].
EI-MS (Basis-Ion ist auch hier m/z - 91. Das Ion m/z 489 beträgt ca. 5 % des Basis-Ions): m/z (%) = 507 (84) [M]+, 489 (100) [M-H2O]+, 474 (30) [M-H2O-CH3], 446 (69), 432 (76). - Hochauflösung: gef. 507.3335, für C32H45NO4 ber. 507.3348. - EI-MS (base ion is here also m / z - 91. The ion m / z 489 is approx. 5% of the base ion): m / z (%) = 507 (84) [M] + , 489 (100) [MH 2 O] + , 474 (30) [MH 2 O-CH 3 ], 446 (69), 432 (76). - High resolution: found 507.3335, for C 32 H 45 NO 4 calc. 507.3348. -
Ferner betrifft die Erfindung ein Verfahren zur Herstellung von Fenalamiden, insbesondere der vorstehend angeführten Fenalamide A, B und/oder C, bei dem man - Myxococcus stipitatus DSM 6259 in einem Medium mit einem Gehalt an Kohlenstoffquellen, Stickstoffquellen und Mineralsalzen in Gegenwart eines Adsorberharzes kultiviert, The invention further relates to a process for the preparation of fenalamides, in particular the fenalamides A, B and / or C mentioned above, in which - Myxococcus stipitatus DSM 6259 cultivated in a medium containing carbon sources, nitrogen sources and mineral salts in the presence of an adsorber resin,
- das Adsorberharz abtrennt und mit Methanol extrahiert,  the adsorber resin is separated off and extracted with methanol,
- aas Eluat zu einem Rohextrakt einengt,  - the eluate is concentrated to a crude extract,
- gegebenenfalls den Rohextrakt in Essigsäureethylester/Wasser aufnimmt, einer Gegenstromzentrifugation unterwirft, die Wasserυhase verwirft und die Esterphase zu einem Rohextrakt einengt,  - if necessary, taking up the crude extract in ethyl acetate / water, subjecting it to countercurrent centrifugation, discarding the water phase and concentrating the ester phase to a crude extract,
- den Rohextrakt der letzten oder der vorletzten Stufe einer Flüssigchromatographie (HPLC) an einer Umkehrphase (RP-18) mit Methanol/Wasser chromatographiert,  - the crude extract of the last or the penultimate stage of liquid chromatography (HPLC) is chromatographed on a reverse phase (RP-18) with methanol / water,
- Fraktionen enthaltend Wirkstoff mit antibiotischer und/oder fungizider Aktivität gewinnt und die Fraktionen einengt, beispielsweise  - Fractions containing active ingredient with antibiotic and / or fungicidal activity wins and the fractions are concentrated, for example
eine Fraktion mit einem Gehalt an Fenalamid A und/oder eine Fraktion mit Fenalamid B und/oder eine Fraktion mit Fenalamid C gewinnt, wobei diese Fraktionen in der angegebenen Reihenfolge anfallen, die Fraktion. en) einengt und  a fraction containing fenalamide A and / or a fraction containing fenalamide B and / or a fraction containing fenalamide C is obtained, these fractions being obtained in the order given, the fraction. en) constricts and
- gegebenenfalls die Flüssigchromatographie mit Fraktionierung und Einengung wiederholt.  - If necessary, the liquid chromatography with fractionation and concentration is repeated.
Zur Herstellung von Cis-Trans-Isomeren der Fenalamide A, B und/oder C bzw. von Fenalamiden einer oder mehrerer der allgemeinen Formeln kann man Fenalamid A, B und/oder C einer Behandlung mit UV-Licht. unterwerfen und die erhaltenen Isomeren chromatographisch auftrennen und gegebenenfalls isolieren. Fenalamide A, B and / or C can be treated with UV light to prepare cis-trans isomers of fenalamides A, B and / or C or of fenalamides of one or more of the general formulas. subject and separate the isomers obtained chromatographically and isolate if necessary.
Ferner betrifft die Erfindung ein therapeutisches Mittel (ausgenommen zur Bekämpfung von Viruserkrankungen), bestehend aus mindestens einem Fenalamid oder enthaltend mindestens ein The invention further relates to a therapeutic agent (except for combating viral diseases) consisting of at least one fenalamide or containing at least one
Fenalamid gegebenenfalls neben einem üblichen Träger und/oder Verdünnungsmittel. Fenalamide if necessary in addition to a conventional carrier and / or diluent.
Dieses Mittel kann zur Bekämpfung von pilzlichen oder parasitären £rκranκungen von Mensen und Tier verwendet werαen. Schließlich betrifft die Erfindung ein Mittel zum Pflanzenschutz für Landwirtschaft, Forstwirtschaft und/oder Gartenbau, This agent can be used to combat fungal or parasitic diseases of canteens and animals. Finally, the invention relates to an agent for crop protection for agriculture, forestry and / or horticulture,
bestehend aus mindestens einem Fenalamid oder enthaltend consisting of or containing at least one fenalamide
mindestens ein Fenalamid gegebenenfalls neben einem üblichen Träger und/oder Verdünnungsmittel. at least one fenalamide, optionally in addition to a conventional carrier and / or diluent.
Nachstehend wird die Erfindung anhand experimenteller Daten näher erläutert. The invention is explained in more detail below on the basis of experimental data.
A. Produktionsbedingungen A. Production conditions
A .1. Produktionsstamm A .1. Production master
Das Bakterium Myxococcus stipitatus gehört zur Ordnung der The bacterium Myxococcus stipitatus belongs to the order of the
Myxobacterales, Unterordnung Cystobacterineae, Familie Myxobacterales, subordination Cystobacterineae, family
Myxococcaceae. Der Produtionsstamm Myxococcus stipitatus Mx s40 wurde im September 1988 an der GBF aus einer Bodenprobe aus dem Jacobsburg State Park bei Wind Gap, Pennsylvania. USA, isoliert, Hinerlegung bei der Deutschen Samlung von Mikroorganismen ( DSM ) unter der Nr. DSM 6259. Myxococcaceae. The production strain Myxococcus stipitatus Mx s40 was obtained at GBF in September 1988 from a soil sample from Jacobsburg State Park near Wind Gap, Pennsylvania. USA, isolated, put down at the German Collection of Microorganisms (DSM) under the number DSM 6259.
A .2. Stammkultur A .2. Root culture
Die Stammkultur erfolgt auf Agarplatten, bevorzugt auf Hefe-Agar (VY/2-agar). Dieses Medium enthält 0.5 % Bäckerhefe, 0.1 % CaCl₂ × H₂O, 0.1μg/l Vitamin B12 und 1.2% Agar. Der pH-Wert wird auf 7.2 eingestellt. Die Platten werden bei 30 ; bebrütet. The stock culture is carried out on agar plates, preferably on yeast agar (VY / 2 agar). This medium contains 0.5% baker's yeast, 0.1% CaCl₂ × H₂O, 0.1μg / l vitamin B12 and 1.2% agar. The pH is adjusted to 7.2. The plates are at 30 ; incubated.
A.3. Morphologische Beschreibung A.3. Morphological description
Die vegetativen Stäbchen sind schlank und haben leicht verjüngte Enden. Sie sind etwa 0,7μm dick und 2 bis 5μm lang. Die Myxosporen sind oval, stark lichtbrechend, etwa 1.1 bis 1.3 × 1.3 bis 1.5 μm groß. Bedingt durch die Gleitbewegung der The vegetative sticks are slender and have slightly tapered ends. They are about 0.7μm thick and 2 to 5μm long. The Myxospores are oval, highly refractive, about 1.1 to 1.3 × 1.3 to 1.5 μm in size. Due to the sliding movement of the
Bakterien, breiten sich die Kolonien rasch über die Kulturplatte aus. Die Schwarmkolonie auf Hefeagar ist dünn, filmartig, rötlich braun. Wie an dem um die Kolonien entstehenden Klärhof zu erkennen, werden die Hefezellen im Medium abgebaut. In älteren Kulturen (1 bis 2 Wochen bei 30' ) werden oft weißliche bis blaß bräunliche Fruchtkörper gebildet. Diese bestehen aus einem weichschleimigen Kopf von etwa 100 bis 200 μm Durchmesser, der oft einem mehr oder weniger langen Stiel von 30 bis 50 μm Durchmesser aufsitzt. Nach mehreren Überimpfungen läßt die  Bacteria, the colonies spread rapidly across the culture plate. The swarm colony on Hefeagar is thin, film-like, reddish brown. The yeast cells in the medium are broken down, as can be seen from the clarification plant that is being built around the colonies. In older cultures (1 to 2 weeks at 30 '), whitish to pale brown fruiting bodies are often formed. These consist of a soft, slimy head of about 100 to 200 μm in diameter, which often sits on a more or less long stem of 30 to 50 μm in diameter. After several vaccinations, the
Tendenz, Fruchtkörper zu bilden, allmählich nach. Bei Tendency to form fruiting bodies gradually. at
Bestrahlung mit ultraviolettem Licht mit einer Wellenlänge von 366 nm zeigt eine einige Tage alte Schwarmkolonie eine gelbe Fluoreszenz. Irradiation with ultraviolet light with a wavelength of 366 nm shows a swarm colony a few days old showing yellow fluorescence.
A.4. Leistungen A.4. Services
Der Stamm Mx s40 produziert Substanzen, nämlich Fenalamide, die das Wachstum von gewissen Pilzen und Bakterien hemmen (vgl. The Mx s40 strain produces substances, namely fenalamides, that inhibit the growth of certain fungi and bacteria (cf.
unten). Die Hemmstoffe können sowohl aus den Zellen wie auch aus dem Kulturüberstand isoliert werden. below). The inhibitors can be isolated both from the cells and from the culture supernatant.
A.5. Produktion der Fenalamide A.5. Fenalamide production
Die Substanzen werden während der logarithmischen bis hin zur stationären Phase produziert. Eine typische Fermentation The substances are produced during the logarithmic to the stationary phase. A typical fermentation
verläuft wie folgt: Ein Fermentor mit 700 1 Arbeitsvolumen wird mit 630 1 Kulturmedium gefüllt (Zusammensetzung: 0.6 % Pepton aus Casein, (tryptisch verdaut) 0.05 % CaCl₂×2H₂O, 0.2% MgSO₄×7H₂O, pH 7.2 mit H₂ SO₄ eingestellt ). Zur Bindung der in das Medium proceeds as follows: A fermentor with 700 1 working volume is filled with 630 1 culture medium (composition: 0.6% peptone from casein, (tryptically digested) 0.05% CaCl₂ × 2H₂O, 0.2% MgSO₄ × 7H₂O, pH 7.2 adjusted with H₂ SO₄). For binding the in the medium
freigesetzten Hemmstoffe wird dem Medium 1 bis 2 % (V/V) eines Adsorberharzes z.B. XAD-1180 zugesetzt. Beimpft wird mit 70 1 einer im gleichen Medium in einem entsprechend kleineren Released inhibitors are the medium 1 to 2% (V / V) of an adsorbent resin e.g. XAD-1180 added. Inoculated with 70 1 one in the same medium in a correspondingly smaller one
Fermentor erzeugten, einen Tag alten Vorkultur. Fermentiert wird bei 30º mit einer Rührgeschwindigkeit von 150 Upm und einer Belüftung von 0.2 V/Vm. Anfängliche Schaumbildung wird durch die Zugabe von 70ml Silikon Antischaum z.B. Tegosipon verhindert. Der pH-Wert wird durchgeregelte Zugabe von 5%iger H₂ SO₄ bei 7.2 gehalten. Die Fermentation wird nach drei Tagen beendet. Fermentor produced a day-old preculture. The fermentation takes place at 30 ° with a stirring speed of 150 rpm and aeration of 0.2 V / Vm. Initial foam formation is prevented by adding 70ml of silicone anti-foam, for example tegosipon. The pH is kept at 7.2 by controlled addition of 5% H₂SO₄. The fermentation is ended after three days.
B. Gewinnung und Charakterisierung B. extraction and characterization
Aufarbeitung eines 700 1-Feπnenters des Staames Mx s40 Refurbishment of a 700 1-finent Mx s40
Die Fermentation wird in Gegenwart von 81 Asorber-Harz XAD-1180 durchgeführt. Gegen Ende der Fermentation wird das Harz vom Medium und den Zellen durch ein Sieb abgetrennt. Das Harz wird in einer Säule mit 3 Bettvolumen Methanol eluiert. Nach Einengen am Rotationsverdampfer werden hieraus 234 g Rohextrakt erhalten. Der Rohextrakt wird in 20 1 Esssigsäureethylester und 20 1 Wasser gelöst und in einer Gegestromzentrifugation verteilt. Die Wasserphase wird verworfen, die Essigesterphase getrocknet und im Wasserstrahlvakuum eingeengt. Ausbeute Essigesterphase: 38 g.  The fermentation is carried out in the presence of 81 Asorber resin XAD-1180. Towards the end of the fermentation, the resin is separated from the medium and the cells through a sieve. The resin is eluted in a column with 3 bed volumes of methanol. After concentrating on a rotary evaporator, 234 g of crude extract are obtained. The crude extract is dissolved in 20 l of ethyl acetate and 20 l of water and distributed in a countercurrent centrifugation. The water phase is discarded, the ethyl acetate phase is dried and concentrated in a water jet vacuum. Yield ethyl acetate phase: 38 g.
Der Rohextrakt wird auf einer präparativen HPLC-Anlage (Prep-bar Trennanlage, Fa. Merck, Darmstadt) auf einer 2-1-Säule Lichroprep RP-18 mit dem Laufmittel (A) Methanol: Wasser = 75: 25 (zunächst 25 min isokratisch) danach mit einem linearen Gradienten (90 min) zum Laufmittel B (Methanol: Wasser= 85:15) chromatographiert. Der Fluß beträgt dabei 120 ml/min, detektiert wird bei λ= 330 nm.  The crude extract is on a preparative HPLC system (Prep-bar separation system, Merck, Darmstadt) on a 2-1 column Lichroprep RP-18 with the eluent (A) methanol: water = 75: 25 (initially 25 min isocratic ) then chromatographed using a linear gradient (90 min) to eluent B (methanol: water = 85:15). The flow is 120 ml / min, detection is carried out at λ = 330 nm.
Die Fenalamide enthaltenden Fraktionen werden zwischen den RetentionsZeiten Rt= 71.5-77.5 min (C), zwischen Rt= 101.5-115 min (A) und zwischen 125.5 und 130.5 min (B.) gesammelt. The fractions containing fenalamides are collected between the retention times R t = 71.5-77.5 min (C), between R t = 101.5-115 min (A) and between 125.5 and 130.5 min (B.).
Nach Einengen werden von A 3.03 g, von B.225 mg und von C 125 mg erhalten, B ist noch mit größeren Mengen an Fettsäuren und deren Glyceriden verunreinigt. Nachtrennungen dieser Fraktionen zur Erfassung der spektroskopischen Daten der Fenalamide erfolgten jeweils an einer Nucleosil C-18-Säule, 7 μAfter concentration, 3.03 g of A, B.225 mg and C 125 mg are obtained, B is still contaminated with larger amounts of fatty acids and their glycerides. Night separations of these fractions to record the spectroscopic data of the Fenalamides were each carried out on a Nucleosil C-18 column, 7 μ
(25 cm × 12.5 mm) mit dem gleichen Laufmitteln. (25 cm × 12.5 mm) with the same solvent.
Beschreibung der Fenalamide Description of fenalamides
Fenalamid A Fenalamid A
HPLC (Säule: Nucleosil-C18, 5μ, 12.5 cm × 4 mm; Laufmittel: Methanol:Wasser= 85/15; Durchfluß: 1.5 ml/ min; Detektion: Diodenarray-System HP 1040A): Retentionszeit Rt.= 6.5 min. - UV (Methanol): λ max (1g ε) = 356 nm (4.77) HPLC (column: Nucleosil-C 18 , 5μ, 12.5 cm × 4 mm; mobile solvent: methanol: water = 85/15; flow rate: 1.5 ml / min; detection: diode array system HP 1040A): retention time R t . = 6.5 min . - UV (methanol): λ max (1g ε) = 356 nm (4.77)
[α]D 25= - 189° (c = 1, in Methanol)[α] D 25 = - 189 ° (c = 1, in methanol)
1H-NMR (600 MHz, CD3OD): δ = 7.24 [t, J= 7.7 Hz, 2H, meta-Protonen am Aromaten], 7.16 [d, J = 7.2 Hz, 2H, ortho-Protonen am Aromaten], 7.14 [t, J = 7.3 Hz, 1H, para-Proton am Aromat], 7.07 [dd, J= 11.0 und 14.5 Hz, 1H, 5-H], 7.02 [d, J = 11.4 Hz, 1H, 3-H], 7.73 [dd, J = 10.6 und 15.1 Hz, 1H, 8-H], 6.59 [dd, J = 11.4 und 14.5 Hz, 1H, 4-H], 6.40 [d, J = 15.1 Hz, 1H, 9-H], 6.14 [m, 2H, 6- und 7-H (durch 2D-H-COSY-Spektren ermittelt), 5.63 [d, J = 9.7 Hz, 1H, 11-H], 5.27 [d, J = 9.6 Hz, 1H, 15-H], 4.09 [dq, J = 5.7 und 12.4 Hz, 1H, 1'-H], 3.85 [d, J = 6.7 Hz, 1H, 13-H], 5.72 [dq, J = 5.8 und 11.0 Hz, 2H, 2'-H2], 2.83 [m, 1H, 12-H], 2.64 [m, 1H, 18Ha] , 2.52 [m, 1H, 18Hb], 2.45 [m, 1H, 16-H], 2.02 [d, J = 0.8 Hz, 3H, 2-CH3], 1.89 [d, J = 1.0 Hz, 3H, 10-CH3], 1.68 [m, 1H, 17-Ha], 1.62 [d, J = 1.2 Hz, 3H, 14-CH3], 1.54 [m, 1H, 17-Hb], 1.23 [d, J = 6.8 Hz, 3H, 1'-CH3], 1.00 [d, J = 1.0 Hz, 3H, 16-CH3], 0.97 [d, JJ = 6.9 Hz, 3H, 12-CH3]. - EI-MS: m/z (%) = 491 (3) [M]+, 473 (6) [M-H2O]+, 455 (2) [M-2 × H2O], 289 (100) [C18H27NO2], 271 (30) [m/z = 289-H2O]. - Hochauflösung: gefunden 491.3381, für C32H45NO3 ber. 491.3399 Fenalamid B 1H-NMR (600 MHz, CD 3 OD): δ = 7.24 [t, J = 7.7 Hz, 2H, meta-protons on the aromatic], 7.16 [d, J = 7.2 Hz, 2H, ortho-protons on the aromatic], 7.14 [t, J = 7.3 Hz, 1H, para-proton on the aromatic], 7.07 [dd, J = 11.0 and 14.5 Hz, 1H, 5-H], 7.02 [d, J = 11.4 Hz, 1H, 3-H ], 7.73 [dd, J = 10.6 and 15.1 Hz, 1H, 8-H], 6.59 [dd, J = 11.4 and 14.5 Hz, 1H, 4-H], 6.40 [d, J = 15.1 Hz, 1H, 9 -H], 6.14 [m, 2H, 6- and 7-H (determined by 2D-H-COZY spectra), 5.63 [d, J = 9.7 Hz, 1H, 11-H], 5.27 [d, J = 9.6 Hz, 1H, 15-H], 4.09 [dq, J = 5.7 and 12.4 Hz, 1H, 1'-H], 3.85 [d, J = 6.7 Hz, 1H, 13-H], 5.72 [dq, J = 5.8 and 11.0 Hz, 2H, 2'-H 2 ], 2.83 [m, 1H, 12-H], 2.64 [m, 1H, 18H a ], 2.52 [m, 1H, 18H b ], 2.45 [m, 1H, 16-H], 2.02 [d, J = 0.8 Hz, 3H, 2-CH 3 ], 1.89 [d, J = 1.0 Hz, 3H, 10-CH 3 ], 1.68 [m, 1H, 17-H a ], 1.62 [d, J = 1.2 Hz, 3H, 14-CH 3 ], 1.54 [m, 1H, 17-H b ], 1.23 [d, J = 6.8 Hz, 3H, 1'-CH 3 ], 1.00 [d, J = 1.0 Hz, 3H, 16-CH 3 ], 0.97 [d, JJ = 6.9 Hz, 3H, 12-CH 3 ]. - EI-MS: m / z (%) = 491 (3) [M] + , 473 (6) [MH 2 O] + , 455 (2) [M-2 × H 2 O], 289 (100) [C 18 H 27 NO 2 ], 271 (30) [m / z = 289-H 2 O]. - High resolution: found 491.3381, for C 32 H 45 NO 3 calc. 491.3399 Fenalamid B
HPLC (Bedingungen wie für Fenalamid A): Rt= 9.6 min . - UV (Methanol) : λmax (1g ε) = 346 nm (4.70). -HPLC (conditions as for fenalamide A): R t = 9.6 min. - UV (methanol): λ max (1g ε) = 346 nm (4.70). -
1H-NMR (600 MHz, [D6]Aceton): δ = 7.24 [t, J = 7.6 Hz, 2H, meta- Protonen am Aromat], 7.16 [d, J = 6.9 Hz, 2H, ortho-Protonen am Aromat], 7.14 [t, J = 7.3 Hz, 1H, para-Proton am Aromat], 6.95 [d, J = 11.4 Hz, 1H, 3-H], 6.60 [dd, J = 11.4 und 14.6 Hz, 1H, 4-H], 6.55 [dd, J = 14.6 und 10.0 Hz, 1H, 5-H], 6.42 [dd, J = 15.3 und 10.0 Hz, 1H, 6-H], 6.41 [d, J = 15.3 Hz, 1H, 7-H], 6.02 [s, 1H, 9-H], 5.47 [d, J = 9.6 Hz, 1H, 11-H], 5.52 [d, J = 9.6 Hz, 1H, 15-H], 4.03 [m, 1H, 1'-H], 3.82 [d, J = 6.0 Hz, 1H, 13-H], 3.61 [m, 2H, 2'-H2], 2.74 [m, 1H, 12-H], 2.61 [ddd, J = 5.5, 10.5 und 14.0 Hz, 1H, 18-Ha], 2.50 [ddd, J = 6.5, 10.0 und 14.0 Hz, 1H, 18-Hb], 2.44 [m, 1H, 16-H], 2.04 [d, J = 1.0 Hz, 3H, 2-CH3], 1.96 [d, J = 1.1 Hz, 3H, 8-CH3], 1.83 [d, J = 1.0 Hz, 3H, 10-CH3], 1.62 [d, J = 1.1 Hz, 3H, 14-CH3], 1.60 [m, 1H, 17-Ha] , 1.51 [m, 1H, 17-Hb], 1.16 [d, J = 6.8 Hz, 3H, 1'-CH3], 0.96 [d, J = 6.5 Hz, 3H, 16-CH3], 0.95 [d, 6.8 Hz, 3H, 12-CH3]. - EI-MS (Die Ionen im Molpeak-Bereich sind nur sehr schwach. Im Gesamtspektrum ist m/z = 91 Basis-Ion. Das Ion m/z 487 beträgt hiervon ca. 5 %, die angegebenen Prozentzahlen beziehen sich nur auf den genannten Massenbereich) m/z (%) = 505 (17) [M]+, 487 (100) [M-H2O]+, 469 (74) [M-2 × H2O], 454 (48) [M-2 × H2O-CH3]. - Hochauflösunσ: gef. 505.3550 für C33H47N03 ber. 505.3555 1 H-NMR (600 MHz, [D 6 ] acetone): δ = 7.24 [t, J = 7.6 Hz, 2H, meta-protons on the aromatic], 7.16 [d, J = 6.9 Hz, 2H, ortho-protons on Aromat], 7.14 [t, J = 7.3 Hz, 1H, para-proton on Aromat], 6.95 [d, J = 11.4 Hz, 1H, 3-H], 6.60 [dd, J = 11.4 and 14.6 Hz, 1H, 4-H], 6.55 [dd, J = 14.6 and 10.0 Hz, 1H, 5-H], 6.42 [dd, J = 15.3 and 10.0 Hz, 1H, 6-H], 6.41 [d, J = 15.3 Hz, 1H, 7-H], 6.02 [s, 1H, 9-H], 5.47 [d, J = 9.6 Hz, 1H, 11-H], 5.52 [d, J = 9.6 Hz, 1H, 15-H], 4.03 [m, 1H, 1'-H], 3.82 [d, J = 6.0 Hz, 1H, 13-H], 3.61 [m, 2H, 2'-H 2 ], 2.74 [m, 1H, 12-H ], 2.61 [ddd, J = 5.5, 10.5 and 14.0 Hz, 1H, 18-H a], 2.50 [ddd, J = 6.5, 10.0 and 14.0 Hz, 1H, 18-H b], 2:44 [m, 1H, 16-H], 2.04 [d, J = 1.0 Hz, 3H, 2-CH 3 ], 1.96 [d, J = 1.1 Hz, 3H, 8-CH 3 ], 1.83 [d, J = 1.0 Hz, 3H, 10-CH 3 ], 1.62 [d, J = 1.1 Hz, 3H, 14-CH 3 ], 1.60 [m, 1H, 17-H a ], 1.51 [m, 1H, 17-H b ], 1.16 [d , J = 6.8 Hz, 3H, 1'-CH 3 ], 0.96 [d, J = 6.5 Hz, 3H, 16-CH 3 ], 0.95 [d, 6.8 Hz, 3H, 12-CH 3 ]. - EI-MS (The ions in the Molpeak range are only very weak. In the overall spectrum, m / z = 91 base ions. The ion m / z 487 is about 5%, the percentages given relate only to the ones mentioned Mass range) m / z (%) = 505 (17) [M] + , 487 (100) [MH 2 O] + , 469 (74) [M-2 × H 2 O], 454 (48) [M- 2 × H 2 O-CH 3 ]. - High resolution: found. 505.3550 for C 33 H 47 N0 3 calc. 505.3555
Fenalamid C Fenalamid C
HPLC (Bedingungen wie für Fenalamid A): >%= 3.8 min. - UV (Methanol): λmax (1g ε) = 328 nm (4.63). -1 H-NMR (300 MHz, CD3OD) : δ = 7.28 [t, J = 7.1 Hz, 2H, metaProtonen am Aromat], 7.20 [d, J = 6.9 Hz, 2H, ortho-Protonen am Aromat], 7.18 [t, J = 7.1 Hz, 1H, para-Proton am Aromat], 6.96 [d, J = 11.0 Hz, 1H, 3-H], 6.60 [m (höherer Ordnung, 2H, 4- und 5-H], 6.64 [m, höherer Ordnung, 3H, 6-, 7- und 8-H], 5.35 [d, J = 9.6 Hz, 1H, 15-H], 4.08 [m, 1H, 1'-H], 3.92 [d, J = 10.4 Hz, 1H, 13-H], 3.62 [d, J = 9.8 Hz, 1H, 11-H], 3.57 [dq, J = 5.6 und 11.6 Hz, 2H, 2'-H2], 2.65 [m, 3H, 12-, 18-Ha und 18 Hb], 2.51 [m, 2H, 18 Hb und 16-H], 2.01 [s, 3H, 2-CH3], 1.69 [s, 3H, 14-CH3], 1.61 [m, 1H, 17-Ha], 1.42 [s, 3H, 10-CH3], 1.33 [m, 1H, 17-Hb], 1.23 [d, J = 7.2 Hz, 3H, l'-CH3], 1.02 [d, J = 6.8 Hz, 3H, 16- CH3], 0.99 [d, J = 7.3 Hz, 3H, 12-CH3]. -HPLC (conditions as for Fenalamid A):>% = 3.8 min. - UV (methanol): λ max (1g ε) = 328 nm (4.63). - 1 H-NMR (300 MHz, CD 3 OD): δ = 7.28 [t, J = 7.1 Hz, 2H, meta protons on the aromatic], 7.20 [d, J = 6.9 Hz, 2H, ortho-protons on the aromatic], 7.18 [t, J = 7.1 Hz, 1H, para-proton on aromatic], 6.96 [d, J = 11.0 Hz, 1H, 3-H], 6.60 [m (higher order, 2H, 4- and 5-H]) , 6.64 [m, higher order, 3H, 6-, 7- and 8-H], 5.35 [d, J = 9.6 Hz, 1H, 15-H], 4.08 [m, 1H, 1'-H], 3.92 [d, J = 10.4 Hz, 1H, 13-H], 3.62 [d, J = 9.8 Hz, 1H, 11-H], 3.57 [dq, J = 5.6 and 11.6 Hz, 2H, 2'-H 2 ] , 2.65 [m, 3H, 12-, 18-H a and 18 H b ], 2.51 [m, 2H, 18 H b and 16-H], 2.01 [s, 3H, 2-CH 3 ], 1.69 [s , 3H, 14-CH 3 ], 1.61 [m, 1H, 17-H a ], 1.42 [s, 3H, 10-CH 3 ], 1.33 [m, 1H, 17-H b ], 1.23 [d, J = 7.2 Hz, 3H, l'-CH 3 ], 1.02 [d, J = 6.8 Hz, 3H, 16-CH 3 ], 0.99 [d, J = 7.3 Hz, 3H, 12-CH 3 ].
EI-MS (Basis-Ion ist auch hier m/z = 91. Das Ion m/z 489 beträgt ca. 5 % des Basis-Ions): m/z (%) = 507 (84) [M]+, 489 (100) [M-H2O]+, 474 (30) [M-H2O-CH3], 446 (69), 432 (76). - Hochauflösung: gef. 507.3335, für C32H45NO4 ber. 507.3348. - EI-MS (base ion is here also m / z = 91. The ion m / z 489 is approximately 5% of the base ion): m / z (%) = 507 (84) [M] + , 489 (100) [MH 2 O] + , 474 (30) [MH 2 O-CH 3 ], 446 (69), 432 (76). - High resolution: found 507.3335, for C 32 H 45 NO 4 calc. 507.3348. -
Handelsbezeichnungen: Trade names:
XAD-1180 (Rohm & Haas; vgl. Anlage) XAD-1180 (Rohm &Haas; see attachment)
Tegosipon (Goldschmidt; vgl. Anlage) Tegosipon (Goldschmidt; see attachment)
Lichroprep RP-18 (Merck; vgl. Anlage) Lichroprep RP-18 (Merck; see attachment)
Nucleosil C-18 (Macherey-Nagel; vgl. Anlage) Nucleosil C-18 (Macherey-Nagel; see attachment)
Diodenarray-System HP 1040A (Hewlett-Packard) C. Wirkung Diode array system HP 1040A (Hewlett-Packard) C. Effect
Fenalamide hemmen das Wachstum von Pilzen und Bakterien, hauptsächlich gram-positiven Bakterien (vgl . Tabelle) Fenalamides inhibit the growth of fungi and bacteria, mainly gram-positive bacteria (see table)
Fenalamid A und MeOH Gesamtextrakte sind mit Fenalamid A and MeOH total extracts are included
20 μg bzw. 20 μl/Testblättchen wirksam gegen: 20 μg or 20 μl / test papers effective against:
Pilze: Mushrooms:
Hemmhof (mm) >10 (<1S) Pythiura debaryanum  Inhibitory zone (mm)> 10 (<1S) Pythiura debaryanum
Fusarium fujikuroi  Fusarium fujikuroi
II Hucor hiemalis  II Hucor hiemalis
II Trichoderma koningii  II Trichoderma koningii
<10 Phytophthora nicotiana  <10 Phytophthora nicotiana
Phytophthora cactorum  Phytophthora cactorum
Rhizopus arrhizus  Rhizopus arrhizus
Botrytis cinerea  Botrytis cinerea
Bakterien; >10 (<15) Micrococcus luteus Bacteria; > 10 (<15) Micrococcus luteus
Nocardia flava  Nocardia flava
Rhodococcus rhodochrous Rhodococcus rhodochrous
<10 Bacillus subtilis <10 Bacillus subtilis
Staphylococcus aureus  Staphylococcus aureus
Micrococcus spp. 1TS 3/1  Micrococcus spp. 1TS 3/1
Sehr schwach wirksam gegen Gram-negative Bakterien: Very weakly effective against Gram-negative bacteria:
Escherichia σoli Tol C Serratia marcescens Escherichia σoli Tol C Serratia marcescens
Escherichia coli AI  Escherichia coli AI

Claims

Patentansprüche 1. Fenalamid der folgenden allgemeinen Formel: Claims 1. Fenalamide of the following general formula:
Figure imgf000017_0001
Figure imgf000017_0001
wobei die gestrichelte Linie zwischen dem C10- und dem C11-Atom eine (weitere) C-C-Bindung oder eine -O-Brücke sein kann und wobei das C8-Atom eine Methylgruppe tragen kann. where the dashed line between the C10 and the C11 atom can be a (further) CC bond or an -O bridge and where the C8 atom can carry a methyl group.
2. Fenaiamid, der folgenden aligemeinen Formel: 2. Fenaiamide, the following general formula:
Figure imgf000018_0001
Figure imgf000018_0001
3. Fenalamid, gekennzeichnet durch einen oder mehrere der folgenden Parameter, 3. Fenalamide, characterized by one or more of the following parameters,
HPLC (Säule: Nucleosil-C18, 5μ, 12.5 cm × 4 mm; Laufmittel: Methanol:Wasser= 85/15; Durchfluß: 1.5 ml/ min; Detektion: Diodenarray-Sysuem HP 1040A): Retenrionszeit Rt= 6.5 min. - UV (Methanol): λ max (1g ε) = 356 nm (4.77) HPLC (column: Nucleosil-C 18 , 5μ, 12.5 cm × 4 mm; mobile phase: methanol:water = 85/15; flow: 1.5 ml/min; detection: diode array system HP 1040A): retention time R t = 6.5 min. - UV (methanol): λ max (1g ε) = 356 nm (4.77)
[α]D 25= - 189° (c = 1, in Methanol) [α] D 25 = - 189° (c = 1, in methanol)
1H-NMR (600 MHZ, CD3OD): δ = 7.24 [t, J= 7.7 Hz, 2H, meta-Protonen am Aromaten], 7.16 [d, J = 7.2 Hz, 2H, ortho-Protonen am Aromaten], 7.14 [t, J = 7.3 Hz, 1H, para-Proton am Aromat], 7.07 [dd, J= 11.0 und 14.5 Hz, 1H, 5-H], 7.02 [d, J = 11.4 Hz, 1H, 3-H], 7.73 [dd, J = 10.6 und 15.1 Hz, 1H, 8-H], 6.59 [dd, J = 11.4 und 14.5 Hz, 1H, 4-H], 6.40 [d, J = 15.1 Hz, 1H, 9-H], 6.14 [m, 2H, 6- und 7-H (durch 2D-H-COSY-Spektren ermittelt), 5.63 [d, J = 9.7 Hz, 1H, 11-H], 5.27 [d, J = 9.6 Hz, 1H, 15-H], 4.09 [dq, J = 5.7 und 12.4 Hz, 1H, 1'-H], 3.85 [d, J = 6.7 Hz, 1H, 13-H], 5.72 [dq, J = 5.8 und 11.0 Hz, 2H, 2'-H2], 2.83 [m, 1H, 12-H], 2.64 [m, 1H, 18Ha], 2.52 [m, 1H, 18Hb], 2.45 [m, 1H, 16-H], 2.02 [d, J = 0.8 Hz, 3H, 2-CH3], 1.89 [d, J = 1.0 Hz, 3H, 10-CH3], 1.68 [m, 1H, 17-HJ, 1.62 [d, J = 1.2 Hz, 3H, 14-CH3], 1.54 [m, 1H, 17-Hb], 1.23 [d, J = 6.8 Hz, 3H, 1'-CH3], 1.00 [d, J = 1.0 Hz, 3H, I6-CH3], 0.97 [d, JJ = 6.9 Hz, 3H, 12-CH3]. - EI-MS: m/z (%) = 491 (3) [M]*, 473 (6) [M-H2O]+, 455 (2) [M-2 × H2O], 289 (100) [C18H27NO2], 271 (30) [m/z = 289-H2O]. - Hochauflösung: gefunden 491.3381, für C32H45NO3 ber. 491.3399 1 H-NMR (600 MHZ, CD 3 OD): δ = 7.24 [t, J = 7.7 Hz, 2H, meta protons on the aromatic], 7.16 [d, J = 7.2 Hz, 2H, ortho protons on the aromatic] , 7.14 [t, J = 7.3 Hz, 1H, para proton on aromatic], 7.07 [dd, J = 11.0 and 14.5 Hz, 1H, 5-H], 7.02 [d, J = 11.4 Hz, 1H, 3- H], 7.73 [dd, J = 10.6 and 15.1 Hz, 1H, 8-H], 6.59 [dd, J = 11.4 and 14.5 Hz, 1H, 4-H], 6.40 [d, J = 15.1 Hz, 1H, 9-H], 6.14 [m, 2H, 6- and 7-H (determined by 2D-H COZY spectra), 5.63 [d, J = 9.7 Hz, 1H, 11-H], 5.27 [d, J = 9.6 Hz, 1H, 15-H], 4.09 [dq, J = 5.7 and 12.4 Hz, 1H, 1'-H], 3.85 [d, J = 6.7 Hz, 1H, 13-H], 5.72 [dq, J = 5.8 and 11.0 Hz, 2H, 2'-H 2 ], 2.83 [m, 1H, 12-H], 2.64 [m, 1H, 18H a ], 2.52 [m, 1H, 18H b ], 2.45 [m , 1H, 16-H], 2.02 [d, J = 0.8 Hz, 3H, 2-CH 3 ], 1.89 [d, J = 1.0 Hz, 3H, 10-CH 3 ], 1.68 [m, 1H, 17- HJ, 1.62 [d, J = 1.2 Hz, 3H, 14-CH 3 ], 1.54 [m, 1H, 17-H b ], 1.23 [d, J = 6.8 Hz, 3H, 1'-CH 3 ], 1.00 [d, J = 1.0 Hz, 3H, I6-CH 3 ], 0.97 [d, JJ = 6.9 Hz, 3H, 12-CH 3 ]. - EI-MS: m/z (%) = 491 (3) [M]*, 473 (6) [MH 2 O] + , 455 (2) [M-2 × H 2 O], 289 (100) [ C18 H27 NO2 ], 271 (30) [m/z = 289- H2 O]. - High resolution: found 491.3381, for C 32 H 45 NO 3 calculated 491.3399
4. Fenalamid, gekennzeichnet durch einen oder mehrere der folgenden Parameter: 4. Fenalamide, characterized by one or more of the following parameters:
HPLC (Säule: Nucleosil-C18, 5μ, 12.5 cm × 4 mm; Laufmittel: Methanol:Wasser= 85/15; Durchfluß: 1.5 ml/ min; Detektion: Diodenarray-System HP 1040A) : Retentionszeit Rt= 9.6 min . -HPLC (column: Nucleosil-C 18 , 5μ, 12.5 cm × 4 mm; mobile phase: methanol: water = 85/15; flow: 1.5 ml/min; detection: diode array system HP 1040A): retention time R t = 9.6 min. -
UV (Methanol): λmax (1g ε) = 346 nm (4.70). -UV (methanol): λ max (1g ε) = 346 nm (4.70). -
1H-NMR (600 MHz. [D5 Aceton): δ = 7.24 [t, J = 7.6 Hz, 2H, metaProtonen am Aromat], 7.16 [d, J = 6.9 Hz, 2H, ortho-Protonen am Aromat], 7.14 [t, J = 7.3 Hz, 1H, para-Proton am Aromat], 6.95 [d, J = 11.4 Hz, 1H, 3-H], 6.60 [dd, J = 11.4 und 14.5 Hz, 1H, 4-H], 6.55 [dd, J = 14.6 und 10.0 Hz, 1H, 5-H], 6.42 [dd, J = 15.3 und 10.0 Hz, 1H, 6-H], 6.41 [d, J = 15.3 Hz, 1H, 7-H], 6.02 [s, 1H, 9-H], 5.47 [d, J = 9. 6 Hz, 1H, 11-H], 5.52 [d, J = 9.6 Hz, 1H, 15-H], 4.03 [m, 1H, 1'-H], 3.82 [d, J = 6.0 Hz, 1H, 13-H], 3.61 [m, 2H, 2'-H2], 2.74 [m, 1H, 12-H], 2.61 [ddd, J = 5.5, 10.5 und 14.0 Hz, 1H, 18-H2], 2.50 [ddd, J = 6.5, 10.0 und 14.0 HZ, 1H, 18-Hb], 2.44 [m, 1H, 16-H], 2.04 [d, J = 1.0 Hz, 3H, 2- CH3], 1.96 [d, J = 1.1 Hz, 3H, 8-CH3], 1.83 [d, J = 1.0 Hz, 3H, 10-CH3], 1.62 [d, J = 1.1 Hz, 3H, 14-CH3], 1.60 [i, 1H, 17-H2], 1.51 [m, 1H, 17-Hb], 1.16 [d, J = 6.3 Hz, 3H, 1'-CH3], 0.96 [d, J = 6.5 Hz, 3H, 16-CH3], 0.95 [d, 6.8 Hz, 3H, 12-CH3]. - HI-MS (Die Ionen im Molpeak-Bereich sind nur sehr schwach. Im Gesamtspektrum ist m/z = 91 Basis-Ion. Das Ion m/z 487 beträgt hiervon ca. 5 %, die angegebenen Prozentzahlen beziehen sich nur auf den genannten Massenbereich) m/z (%) = 505 (17) [M]", 487 (100) [M-H2O]-, 469 (74) [M-2 × H2O], 454 (48) [M-2 × H2O-CH3]. - Hochauflösunσ: gef. 505.3550 für C33H47NO3 ber. 505.3555 1 H-NMR (600 MHz. [D 5 acetone): δ = 7.24 [t, J = 7.6 Hz, 2H, metaprotons on the aromatic], 7.16 [d, J = 6.9 Hz, 2H, ortho-protons on the aromatic], 7.14 [t, J = 7.3 Hz, 1H, para proton on aromatic], 6.95 [d, J = 11.4 Hz, 1H, 3-H], 6.60 [dd, J = 11.4 and 14.5 Hz, 1H, 4-H ], 6.55 [dd, J = 14.6 and 10.0 Hz, 1H, 5-H], 6.42 [dd, J = 15.3 and 10.0 Hz, 1H, 6-H], 6.41 [d, J = 15.3 Hz, 1H, 7 -H], 6.02 [s, 1H, 9-H], 5.47 [d, J = 9. 6 Hz, 1H, 11-H], 5.52 [d, J = 9.6 Hz, 1H, 15-H], 4.03 [m, 1H, 1'-H], 3.82 [d, J = 6.0 Hz, 1H, 13-H], 3.61 [m, 2H, 2'-H 2 ], 2.74 [m, 1H, 12-H] , 2.61 [ddd, J = 5.5, 10.5 and 14.0 Hz, 1H, 18-H 2 ], 2.50 [ddd, J = 6.5, 10.0 and 14.0 HZ, 1H, 18-H b ], 2.44 [m, 1H, 16 -H], 2.04 [d, J = 1.0 Hz, 3H, 2-CH 3 ], 1.96 [d, J = 1.1 Hz, 3H, 8-CH 3 ], 1.83 [d, J = 1.0 Hz, 3H, 10 -CH 3 ], 1.62 [d, J = 1.1 Hz, 3H, 14-CH 3 ], 1.60 [i, 1H, 17-H 2 ], 1.51 [m, 1H, 17-H b ], 1.16 [d, J = 6.3 Hz, 3H, 1'-CH 3 ], 0.96 [d, J = 6.5 Hz, 3H, 16-CH 3 ], 0.95 [d, 6.8 Hz, 3H, 12-CH 3 ]. - HI-MS (The ions in the molar peak area are only very weak. In the overall spectrum m / z = 91 base ion. The ion m / z 487 is approx. 5% of this, the percentages given only refer to the one mentioned Mass range) m/z (%) = 505 (17) [M]", 487 (100) [MH 2 O]-, 469 (74) [M-2 × H 2 O], 454 (48) [M- 2 × H 2 O-CH 3 ]. - High resolution: found 505.3550 for C 33 H 47 NO 3 calculated 505.3555
5. Fenalamid, gekennzeichnet durch einen oder mehrere der folgenden Parameter: 5. Fenalamide, characterized by one or more of the following parameters:
HPLC (Säule: Nucleosil-C18, 5μ, 12.5 cm × 4 mm; Laufmittel: Methanol:Wasser= 85/15; Durchfluß: 1.5 ml/ min; Detektion: Diodenarray-System HP 1040A): Retentionszeit Rt= 3.8 min. - HPLC (column: Nucleosil-C 18 , 5μ, 12.5 cm × 4 mm; mobile phase: methanol:water = 85/15; flow: 1.5 ml/min; detection: diode array system HP 1040A): retention time R t = 3.8 min. -
UV (Methanol) : λmax (1g ε) = 328 nm (4.63). -UV (methanol): λ max (1g ε) = 328 nm (4.63). -
1H-NMR (300 MHz, CD3OD): δ = 7.28 [t, J = 7.1 Hz, 2H, metaProtonen am Aromat], 7.20 [d, J = 6.9 Hz, 2H, ortho-Protonen am Aromat], 7.18 [t, J = 7.1 Hz, 1H, para-Proton am Aromat], 6.96 [d, J = 11.0 Hz, 1H, 3-H], 6.60 [m (höherer Ordnung, 2H, 4- und 5-H], 6.64 [m, höherer Ordnung, 3H, 6-, 7- und 8-H], 5.35 [d, J = 9.6 Hz, 1H, 15-H], 4.08 [m, 1H, 1'-H], 3.92 [d, J = 10.4 Hz, 1H, 13-H], 3.62 [d, J = 9.8 Hz, 1H, 11-H], 3.57 [dq, J = 5.6 und 11.6 Hz, 2H, 2'-H,], 2.65 [m, 3H, 12-, 18-Hm und 18 Hb] , 2.51 [m, 2H, 18 H5 und 16-H], 2.01 [s, 3H, 2-CH3], 1.69 [s, 3H, 14-CH3], 1.61 [m, 1H, 17-Ha], 1.42 [s, 3H, 10-CH3], 1.33 [m, 1H, 17-H5], 1.23 [d, J = 7.2 Hz, 3H, 1'-CH3], 1.02 [d, J = 6.8 Hz, 3H, 16-CH3], 0.99 [d, J = 7.3 Hz, 3H, 12-CH3]. - 1 H-NMR (300 MHz, CD 3 OD): δ = 7.28 [t, J = 7.1 Hz, 2H, metaprotons on the aromatic], 7.20 [d, J = 6.9 Hz, 2H, ortho-protons on the aromatic], 7.18 [t, J = 7.1 Hz, 1H, para proton on aromatic], 6.96 [d, J = 11.0 Hz, 1H, 3-H], 6.60 [m (higher order, 2H, 4- and 5-H], 6.64 [m, higher order, 3H, 6-, 7- and 8-H], 5.35 [d, J = 9.6 Hz, 1H, 15-H], 4.08 [m, 1H, 1'-H], 3.92 [ d, J = 10.4 Hz, 1H, 13-H], 3.62 [d, J = 9.8 Hz, 1H, 11-H], 3.57 [dq, J = 5.6 and 11.6 Hz, 2H, 2'-H,], 2.65 [m, 3H, 12-, 18-H m and 18 H b ], 2.51 [m, 2H, 18 H 5 and 16-H], 2.01 [s, 3H, 2-CH 3 ], 1.69 [s, 3H, 14-CH 3 ], 1.61 [m, 1H, 17-H a ], 1.42 [s, 3H, 10-CH 3 ], 1.33 [m, 1H, 17-H 5 ], 1.23 [d, J = 7.2 Hz, 3H, 1'-CH 3 ], 1.02 [d, J = 6.8 Hz, 3H, 16-CH 3 ], 0.99 [d, J = 7.3 Hz, 3H, 12-CH 3 ]. -
EI-MS (Basis-Ion ist auch hier m/z = 91. Das Ion m/z 489 beträgt ca. 5 % des Basis-Ions): m/z (%) = 507 (84) [M]+, 489 (100) [M- H2O+, 474 (30) [M-H2O-CH3], 446 (69), 432 (76). - Hochauflösung: gef. 507.3335, für C32H45NO4 ber. 507.3348. - EI-MS (base ion here is also m/z = 91. The ion m/z 489 is approx. 5% of the base ion): m/z (%) = 507 (84) [M] + , 489 (100) [M-H 2 O + , 474 (30) [MH 2 O-CH 3 ], 446 (69), 432 (76). - High resolution: found. 507.3335, for C 32 H 45 NO 4 calc. 507.3348. -
6. Verfahren zur Herstellung von Fenalamiden, insbesondere gemäß einem der Ansprüche 1 bis 5, dadurch gekennzeichnet, daß man 6. Process for the production of fenalamides, in particular according to one of claims 1 to 5, characterized in that
- Myxococcus stipitatus DSM 6259 in einem Medium mit einem Gehalt an Kohlenstoffquellen. Stickstoffquellen und Mineralsalzen in Gegenwart eines Adsorberharzes kultiviert, - Myxococcus stipitatus DSM 6259 in a medium containing carbon sources. Nitrogen sources and mineral salts cultivated in the presence of an adsorber resin,
- das Adsorberharz abtrennt: und mit Methanoi extraniert. - the adsorber resin is separated: and extracted with methane.
- das Eluat zu einem Rohextrakt einengt, - the eluate is concentrated to a crude extract,
- gegebenenfalls den Rohextrakt in Essigsäureethyiester/Wasser aufnimmt, einer Gegenstromzentrifugation unterwirft, die Wasserpnase verwirft und die Esterphase zu einem Rohextrakt einengt, - optionally takes up the crude extract in ethyl acetate/water, subjects it to countercurrent centrifugation, discards the water vapor and concentrates the ester phase to form a crude extract,
- den Rohextrakt der letzten oder der vorletzten Stufe einer Flüssigchromatographie (HPLC) an einer Umkehrphase (RP-18) mit Methanol/Wasser chromatographiert, - the crude extract of the last or penultimate stage of liquid chromatography (HPLC) is chromatographed on a reverse phase (RP-18) with methanol/water,
- Fraktionen enthaltend Wirkstoff mit antibiotiscner und/oder fungizider Aktivität gewinnt und die Fraktionen einengt, insbesondere eine Fraktion mit einem Gehalt an Fenalamid gemäß Anspruch 4 und/oder eine Fraktion mit Fenalamid gemäß Anspruch 5 und/oder eine Fraktion mit Fenalamid gemäß Anspruch fi - Fractions containing active ingredient with antibiotic and / or fungicidal activity are obtained and the fractions are concentrated, in particular a fraction containing fenalamide according to claim 4 and / or a fraction with fenalamide according to claim 5 and / or a fraction with fenalamide according to claim fi
gewinnt, wobei diese Fraktionen in der angegebenen Reihenfolge anfallen, die Fraktion. en) einengt und The faction wins, with these fractions occurring in the order listed. en) narrows and
- gegebenenfalls die Flüssigchromatographie mit Fraktionierung und Einengung wiederholt. - if necessary, repeat the liquid chromatography with fractionation and concentration.
7. Verfahren zur Herstellung von Cis-Trans-Isomeren von Fenalamiden gemäß Anspruch 3. 4 und/oder 5 oder von Fenalamiden einer oder mehrerer der allgemeinen Formeln gemäß Anspruch 1 oder 2, dadurch gekennzeichnet, daß man ein Fenalamid gemäß Anspruch 3, 4 und/oder 5 einer Behandlung mit UV-Licht unterwirft und die erhaltenen Isomeren chromatographisch auftrennt und gegebenenfalls isoliert. 7. Process for the preparation of cis-trans isomers of fenalamides according to claim 3, 4 and / or 5 or of fenalamides of one or more of the general formulas according to claim 1 or 2, characterized in that a fenalamide according to claims 3, 4 and / or 5 is subjected to treatment with UV light and the isomers obtained are separated by chromatography and, if necessary, isolated.
5. Therapeutisches Mittel (ausgenommen zur Bekämpfung von Viruserkrankungen), bestehend aus mindestens einem Fenalamid gemäß einem der Ansprüche 1 bis 5 oder enthaltend mindestens ein Fenalamid gemäß einem der Ansprüche 1 bis 5 gegebenenfalls neben einem üblichen Träger und/oder Verdünnungsmittel. 5. Therapeutic agent (except for combating viral diseases), consisting of at least one fenalamide according to one of claims 1 to 5 or containing at least one fenalamide according to one of claims 1 to 5, optionally in addition to a usual carrier and / or diluent.
9. Therapeutisches Mittel nach Anspruch 8 zur Bekämpfung von pilziichen oder parasitären Ernrankungen von Mensch und Tier. 9. Therapeutic agent according to claim 8 for combating fungal or parasitic diseases of humans and animals.
10. Mittel zum Pflanzenschutz für Landwirtschaft, Forstwirtschaft und/oder Gartenbau, bestehend aus mindestens einem Fenalamid gemäß einem der Ansprüche 1 bis 5 oder enthaltend mindestens ein Fenalamid gemäß einem der Ansprüche 1 bis 5 gegebenenfalls neben einem üblichen Träger und/oder Verdünnungsmittel. 10. Plant protection agent for agriculture, forestry and/or horticulture, consisting of at least one fenalamide according to one of claims 1 to 5 or containing at least one fenalamide according to one of claims 1 to 5, optionally in addition to a usual carrier and/or diluent.
PCT/EP1991/002480 1990-12-24 1991-12-20 Phenylpolyenamides, process for producing the same and agents WO1992011231A1 (en)

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CN110042061A (en) * 2019-04-16 2019-07-23 浙江工业大学 High yield gibberellin GA3Gibberella fujikuroi mutant strain and its application

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Title
Chemical Abstracts, Band 100, no. 23, 4 Juni 1984, (Columbus, Ohio, US), Rolf Jansen et al : "Antibiotics from gliding bacteria. XVIII. The absolute configuration of myxalamide A and B ", siehe Seite 558, Zusammenfassung 191623w, & Liebigs Ann. Chem. 1984, 1(), 78- 8 *
The Journal of Antibiotics, Band. 36, Nr. 9, September 1983 K. Gerth et al: "The myxalamids, new antibiotics from myxococcus xanthus (myxobacterales). 1. Production, physico-chemical and biological properties, a", *
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CN110042061A (en) * 2019-04-16 2019-07-23 浙江工业大学 High yield gibberellin GA3Gibberella fujikuroi mutant strain and its application
CN110042061B (en) * 2019-04-16 2020-08-21 浙江工业大学 High yield gibberellin GA3Gibberella fujikuroi mutant strain and application thereof

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