FI93014B - Sorafen, a method for the preparation thereof and its use for the control of plant diseases - Google Patents

Description

9301 4

The present invention relates to a macrocyclic compound of the formula I, to a process for its preparation and to its use for controlling plant diseases and to plant microbicidal substances containing this compound as active ingredient.

? R

/, ν /, ν / 'Χ / π3 P i23 lo 8j · “ÖCHa y en 3 \ n y \ / 2o / I»! 1: i, / VY'W \, * '\ / rk 5 · In this formula, R represents methyl when there is a double bond at the 9,10 position (= compound Ia), or R represents hydrogen when the simple position at the 9,10 position is bond (= compound Ib).

Hereinafter, Compound Ia is also referred to as "sorafen A" and Compound Ib as "sorafen B". Formula I in principle also comprises isomeric structures and mixtures thereof and an arbitrary mixture of compounds Ia and Ib. Sorafen A is preferred.

The compounds of this structural class are new. There is no reference in the literature to such a structure.

The compound of formula I is prepared by microbiological culturing of Sorangium (Polyanqium) cellulosum strain "So: ce 26". This position was deposited on 5 March 1987 with the National Collection of Industrial and Marine Bacteria (NCIB ·), Torrey Research Station, Aberdeen, United Kingdom, under number NCIB 12 411 in accordance with the Budapest Treaty 2 93014. Sorangium cellulosum belongs to the family Polyangiaceae of the Sorangineae subfamily of the order Myxobacteriales.

The microorganism "So ce 26" was isolated in 1980 from a sample of goat manure collected in 1978 in Djerba, Tunisia. "So ce 26" itself, or mutants or recombinants thereof, which likewise produce compounds of formula I, are also a subject of this invention. The microorganism Sorangium "So ce 26" can be cultured according to conventional biological methods, e.g. in shake cultures or fermenters on culture media. The fermentation temperature is generally 10 ° to 35 ° C and preferably about 30 ° to 32 ° C, the pH is 6-8, especially 7.4. The process takes place aerobically and under sterile conditions.

The composition of the culture medium can vary very much. A source of carbon and nitrogen and a source of inorganic elements, including P, S, Mg, K, Ca, must be available for the nutrients to be substantially assimilated.

In the fermentation process, glucose, starch or cellulose are preferably used as the C source, in addition to this, glycerol, acetic acid and others. Suitable N-sources are NH4, NO3 or also peptones. An organic compound as an N source cannot be the only C source and energy source for fermentation at the same time.

Suitable mineral salts are the chlorides, nitrates, sulphates, carbonates and phosphates of the elements Na, K, NH4, Mg and Ca, in addition to which Fe, Cu, Mn, Zn, Co or other trace elements can be present.

The microorganism culture is used in a shaking culture or fermentor at 0.1 to 7% (v / v), especially 0.5 to 5% (v / v). The reaction time at about 30 ° C is about 2-7 days, 3 93014 less often 10 days. For larger batches, smaller precultures are suitably fermented initially.

The use of "So ce 26" is also possible in immobilized form, e.g. in the form of cells attached to a support in alginate.

It will be appreciated that the method of this invention is not limited to the use of strain "So ce 26" NCIB 12411, but may use other natural or laboratory-produced artificial mutants or recombinants capable of producing a compound of formula I. Methods for making mutants and recombinants of a microorganism are known and familiar to those skilled in the art.

Thus, the invention relates to a process for the preparation of a macrocyclic compound of the formula I by culturing the microorganism Sorangium (Polyanqium) cellulosum "So ce 26" (NCIB 12 411) or a mutant or recombinant thereof in an aqueous culture medium, which is also capable of producing compounds of the formula I. In a narrower sense, the method is characterized by culturing the microorganism Sorangium (Polyangium) cellulosum "So ce 26" (NCIB 12 411) in a culture medium containing at least one assimilable C-source and N-source and corresponding inorganic salts, 10 ° to 35 At 0 ° C in the presence or absence of adsorber resin, then the culture broth or vast, filtered adsorber resin is extracted with a suitable solvent phase, the resulting solution is evaporated and the residual residue is purified, if desired, by chromatography and / or recrystallization.

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Stock culture and morphological description:

Stock cultures are maintained on VY / 2 agar (0.5% baking yeast according to fresh weight, 0.1% CaCl2, 1.5% agar, pH 7.2) or filter paper on ST21 agar (0.1% KNO 3, 0.1% MgSO4.7H2O, 0.1% CaCl2, 0.1% Κ2ΗΡ04, 0.01% MnSO4.7H2O, 0.02% FeCl3, 0.002% yeast extract, standard trace element solution, 1% agar). The plates are incubated at 30 ° C.

On filter paper on mineral salt agar or on yeast agar (VY / 2 agar), the strain forms large mass colonies with several yellow-orange - black-brown spores. These consist of small sporangiols with a diameter of 15 to 20 nm which are more or less in large piles tightly packed. Heaps are usually 50 to 150 microns in diameter. Vegetative rod bacteria have a shape typical of Sorangium: relatively dense, dark under a phase contrast microscope,. cylindrical rod bacteria with broadly rounded ends having an average length of about 3 to 6 microns and a thickness of about 1 microns After long adaptation to growth in a liquid medium, the strain grows as a homogeneous cell suspension.

Biological characterization of strain "So ce 26" (NCIB 12 411) glucose degradation: positive cellulose degradation: positive starch degradation: positive NH4 as N source: positive V NO3 as N source: positive

Strain So ce 26 produces two chemically related compounds Ia and Ib that inhibit the growth of several fungi. These

Il.

5,9014 macrocyclic compounds can be isolated from both cells and culture supernatant.

From the aqueous fermentation broth, compound I can be extracted lipophilically with organic solvents, e.g. ketones such as methyl ethyl ketone, cyclohexanone, medium chain alcohols such as isobutanol, pentanol, pentanol, hexanol, ethyl acetate, acetic acid-ethyl-C 6 -alkyl, , isobutyl acetate, etc.), hydrocarbons or chlorinated hydrocarbons such as hexane, toluene, dichloromethane, 1,2-dichloroethane, chlorobenzene, dichlorobenzene, etc.

From the filtered cell cake, compound I can be easily extracted with alcohols or ketones (e.g. methanol, ethanol, acetone, methyl ethyl ketone).

The compound of the formula I in question can then be recovered from the extract in question, which can be separated by fractional crystallization or other separation methods (e.g. column chromatography) into both components Ia and Ib.

Fermentation to prepare a compound of formula I is preferably carried out in the presence of an adsorber resin from which the desired product is recovered. Suitable adsorber resins are, above all, neutral organic polymeric substances, in particular non-ionic hydrophobic adsorber resins which are suitable for lipophilic extraction. To these are bound the compound of formula I almost quantitatively. Examples of such resins are semi-polar acrylic ester resins, non-polar polystyrene / divinylbenzene resins and especially crosslinked polystyrene. Such resins are added in an amount of 0.1 to 5% (v / v) of the fermentation volume of ferr 93014, in particular 0.5 to 2% (v / v). Activated carbon can also be used.

A technically highly preferred variant for the preparation of a compound of formula I is based on the fermentation of "So ce 26" (NCIB 12 411) under the conditions described above in the presence of a polystyrene resin (e.g. XAD-1180, Rohm and Haas), preferably in filterable form (granules). or granules). At the end of the fermentation, the resin is filtered off, washed with water and treated with methanol or ethanol. The alcoholic extract is evaporated, compound Ia (= sorafen A) which crystallizes on addition of diethyl ether is filtered off and the filtrate is evaporated to recover compound Ib (= sorafen B) and, if desired, purified by chromatography.

The invention relates to a compound of formula I in pure form or in crystallized form. However, the invention also relates to those derived from fermentation. biomass, crude extracts or adsorber resins containing a compound of formula I which can be used as such or in a further modified form for the control of plant diseases. Biomass can be further used or sold as ground or compressed dry matter ("cake").

The disclosed manufacturing methods, including all sub-steps, are the subject of this invention.

surprisingly, it has now been found that the compounds of the formula I have a very advantageous biocidal spectrum for phytopathogenic microorganisms, in particular against fungi, for practical purposes. They have very advantageous curative, systemic and especially preventive properties and are used to protect various crops.

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The active compounds of the formula I can be used to prevent or eradicate pests present in plants or parts of plants (fruits, flowers, leaves, stems, tubers, roots) of various useful crops, so that later growing parts of the plant are also spared from phytopathogenic microorganisms.

As microbicides, the active compounds of the formula I are active, e.g. . In addition, they are effective against the classes Ascomycetes (e.g., especially Venturis and Erysiphe, further Podosphaera, Monilinia, Uncinula), and Oomycetes (e.g., Phytophora, Plasmospora). The compounds of the formula I can further be used as a pickling agent for the treatment of seeds (fruits, tubers, grains) or plant cuttings in order to protect them against fungal infections and against phytopathogenic fungi East.

The invention also relates to substances which contain compounds of the formula I as active ingredient components, in particular plant protection agents, and to their use in the agricultural sector or in related fields.

The invention likewise relates to a process for the treatment of plants, which process is characterized in that new compounds of the formula I or new substances containing them are applied.

For the purposes of the present invention, the following plant species are suitable as target crops for the plant protection use disclosed herein: cereals (wheat, barley, rye, oats, rice, β 9301 4 maize, sorghum and the like), beets (sugar and fodder beets), , stone and berry fruits (apples, pears, plums, peaches, almonds, cherries, strawberry, raspberry and blackberry), legumes (beans, lentils, peas, soybeans), oilseeds (rapeseed, mustard, poppy, olive, sunflower, coconut, castor, cocoa, peanuts), cucumbers (pumpkin, cucumbers, melons), fiber plants (cotton, flax, hemp, jute), citrus fruits (oranges, lemons, grapefruits, mandarins), vegetables (spinach, lettuce, asparagus, cabbage, cabbage, cabbage) onion, tomato, potato, paprika), bay plants (avocado, cinnamon, camphor) or plants such as tobacco, nuts, coffee, pineapple, sugar cane, tea, pepper, vines, hops, bananas and natural rubber plants, and ornamental plants. plants (Compositeae). However, this list is by no means restrictive.

The active compounds of the formula I are usually used in the form of compositions and can be applied simultaneously or sequentially with the other active compounds to the surface or plant to be treated. These other active substances can be fertilizers, trace elements or other substances which affect plant growth. However, they can also be selective herbicides, insecticides, fungicides, bactericides, nematicides, molluscicides or mixtures of several such preparations, together with possibly other carriers, surfactants or other aids which facilitate the application in the art.

Suitable carriers and additives can be solid or liquid and correspond to substances which are suitable in the art of preparation, such as, for example, natural or organic substances.

Il: 9 93014 generated minerals, solvents, dispersants, wetting agents, fixing agents, thickeners, binders or fertilizers.

One preferred method of applying an active substance according to the invention or an agrochemical substance containing at least one such active substance is to apply it to the press (leaf application). The number of applications and the amount of application then depend on the disease pressure of the corresponding pathogen (fungal species). The active substances according to the invention can also penetrate the plant through the roots through the soil (systemic effect) by wetting the plant growth site with a liquid preparation or by applying the substances in solid form to the soil, e.g. in the form of granules (soil application). This granulate or similar powder may be a dry mass of biomass from the fermenter or an adsorber resin filtered from the fermentation broth and containing the active ingredients of the formula I. The compounds of the formula I can also be applied to the seeds by coating the seeds in a liquid preparation of the active ingredient or by coating them with a solid preparation.

The compounds according to the invention are then used in unmodified form or, preferably, in combination with customary auxiliaries in the preparation technique. For this purpose, they are suitably treated in a known manner, e.g. as emulsion concentrates, brushable pastes, directly sprayable or dilutable solutions, dilute emulsions, spray powders, soluble powders, dusts, granules, encapsulating e.g. in polymeric substances. The methods of application, such as spraying, spraying, dusting, application, brushing or irrigation, and the quality of the substances are chosen according to the aims pursued and the conditions prevailing. Suitable application rates are generally from 10 g to 2 kg of active substance (AA) / ha, preferably from 50 g to 500 g of AA / ha.

Formulations, i. substances containing an active ingredient of the formula I and optionally a solid or liquid additive are prepared in a known manner.

Suitable solvents are: aromatic and aliphatic hydrocarbons, such as xylene mixtures, cyclohexane or paraffins, alcohols and glycols, and their ethers and esters, such as ethanol, ethylene glycol, ethylene glycol monomethyl or ethyl ether, acetic acid ester, acetic acid ester. such as cyclohexanone, strongly polar solvents such as N-methyl-2-pyrrolidone, dimethyl sulfoxide or dimethylformamide, and optionally epoxidized vegetable oils such as epoxidized coconut oil or soybean oil, or water.

As solid carriers used, for example, in dusts and dispersible powders, natural stone flours, such as calcite, talc, kaolin, montmorillonite or attapulgite, are generally used. Highly dispersed silicic acid or highly dispersed absorbent polymers can also be added to improve physical properties. Granular adsorbent granulate carrier. suitable porous types are, for example, pumice, crushed brick, sepiolite or bentonite, and non-sorptive carriers are, for example, calcite or sand. In addition to this, several inorganic pre-granulated substances can be used, such as, in particular, dolomite or ground plant residues.

Suitable surfactants are, depending on the nature of the active compound of the formula I to be prepared, non-ionogenic, cationic and / or anionic surfactants having good emulsifying, dispersing and wetting properties. Surfactants also mean mixtures of tensides.

Suitable anionic surfactants can be both so-called water-soluble soaps and water-soluble synthetic surfactants.

Most often, however, so-called synthetic surfactants are used, in particular alkanesulfonates, fatty alcohol sulfates, sulfonated benzimidazole derivatives or alkyl sulfonates.

Suitable nonionic surfactants are polyglycolether derivatives of aliphatic or cycloaliphatic alcohols, saturated or unsaturated fatty acids and alkylphenols which contain from 3 to 30 glycol ether groups and from 8 to 20 carbon atoms in the alkylene alkyl group (in the aliphatic) hydrocarbon residue.

Other examples of nonionic surfactants include nonylphenol polyethoxyethanols, castor oil polyglycol ethers, polypropylene-polyethylene oxide addition products, tributylphenoxypolyethylene ethanol, polyethylene glycol and octylphenoxypolyethoxyethanol.

Fatty acid esters of polyoxyethylene sorbitan, such as polyoxyethylene sorbitan trioleate, are also suitable.

Other surfactants conventional in the art are also known to those skilled in the art or can be obtained from the relevant professional literature.

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Agrochemical preparations generally contain 0.1 to 95% of the active compound of the formula I, 99.9 to 5% of a solid or liquid additive and 0 to 25% of a surfactant.

Concentrated substances are preferred as a commodity, while diluted substances are generally used by the final consumer.

The agents may also contain other additives such as stabilizers, antifoams, viscosity modifiers, binders, binders and fertilizers or other active ingredients to achieve a special effect.

The following examples illustrate the invention without, however, limiting it in any way (percentages and parts are by weight, temperatures are in degrees Celsius).

Preparation Examples:

Example H-1; Preparation of a compound of formula I A 70 liter fermentor (Giovanola, Monthey, Switzerland) is filled with 60 l of medium (0.1% peptone, 0.5% glucose, 0.05% CaCl 2 · 2H 2 O, 0.05% MgSO 4). 4.71 ° C, pH 7.4). Inoculate 10 l of pre-culture (160 rpm, 30 ° C) grown in a shake flask in the same medium. It is then fermented at 32 ° C with a stirring speed of 500 rpm and aeration of 0.5 Nm 4 per hour. Fermentation takes about 7 days. The compound of formula I is produced during a logarithmic-stationary growth phase. It is enriched partly in cells, partly in culture supernatant.

The fermentation broth is then shaken 5 times with 2 liters of ethyl acetate each time for 5 minutes.

13 9301 4 for. The combined extracts are washed twice with water and evaporated in vacuo. The residual dark oil is chromatographed on silica gel (Lichroprep Si60, Merck, 25-40 et seq. Propellant: dichloromethane / acetone in steps 97/3 to 80/20).

The fractions containing compound Ia (= sorafen A) are re-evaporated and chromatographed a second time through silica gel (Lichroprep Si60, Merck, 15-25 et seq. Propellant: dichloromethane / acetone 97/3). The sorafen A thus obtained crystallizes from diethyl ether. Yield: 220 mg.

The fractions containing Ib (= sorafen B) are re-evaporated and chromatographed on the same type of silica gel (15-25) with dichloromethane / acetone (gradient: 90/10-85/15). Fine purification is performed by a third chromatography on a crosslinked dextran gel, such as Sephadex LH-20 (Pharmacia) with methanol. Yield: 100 mg.

• I

‘Example H-2: Preparation of a compound of formula I in the presence of ad sorber i resins

The process is carried out in a fermentation volume of 300 liters by adding 0.5% (v / v) of adsorber resin XAD-1180 (Rohm and Haas). The process conditions for this enlarged batch otherwise correspond to the conditions for Example H-1.

At the end of the fermentation, the polymer support is screened off, rinsed on a glass column, washed with 3 column volumes of water and eluted with 2 column volumes of methanol. The eluate is evaporated to dryness in vacuo and chromatographed on silica gel (Lichroprep Si60, 25-40 [mu] m, Merck, propellant: dichloromethane / acetone 97/3-80 / 20).

14 9301 4 a) The fraction containing sorafen A is evaporated and chromatographed on silica gel (Lichoprep Si60, 15-25 μm, Merck. Propellant: dichloromethane / acetone 97/3).

Optionally, after treatment of the polymeric support with methanol, the methanolic solution can be carefully evaporated without chromatography, whereby the desired sorafen A crystallizes after the addition of diethyl ether.

The product can be recrystallized from diethyl ether. Yield: 1 g of sorafen A.

b) The fraction containing sorafen B is rechromatographed on silica gel (Lichroprep Si60, 15-25 μm, Merck, propellant gradient: dichloromethane / acetone 90/10-85/15). Fine purification is performed by chromatography on Sepha-dex Ifl-20 (Pharmacia, propellant: methanol), yield: 0.6 g ·

Chemical characterization of sorafen A (= comp. Ia) Sum formula: C29H44O8. Mg: 520.

Colorless crystals of diethyl ether, m.p. 101 ° C.

[α] 25 D = -131 ° (MeOH, c = 1) UV (MeOH) λmax <lo9ε> = 317 sh (2.03); 267 sh (2.47); 2.63 (2.59); 2.57 (2.68); 251 (2.66); 245 sh (2.66); 228 sh (2.84); 215 (3.72); 207 (3.94) IR (KBr): 975, 992, 1021, 1045, 1071, 1102, 1152, 1187, 1230, 1255, 1272, 1380, 1451, 1708 (sh), 1729, 2854 (sh), 2923 cm "1.

MS FAB (neg. Ion) = 519 (M-H) -: EI (70ev) = 520

High Resolution: Calc .: 520.3036 Found: 520.3020 DC (DC-Aluminum Foil 6OF254, Merck, propellant: dichloromethane / -153,9014 acetone 90/10) Rp = 0.5 (detection by spraying with anisaldehyde / sulfuric acid reagent and heating to 120 ° C) HPLC (column: 4 x 250 mm Nucleosil 100-7 C ^ g, Macherey Nagel, flow 1.5 ml / min., propellant: methanol / water 70/30 in 20 minutes linearly to 100 / 0) = 11.8 min.

Chemical characterization of sorafen B (= comp. Ib)

Total formula: C28H44 ° 8 * Mg: 508.

[a] 25d. -57 ° (MeOH, c = 1) UV (MeOH) (log ε) = 26 3 sh (2.98); 256 sh (3.11); 247 sh (3.36); 238 (3.42); 215 sh (3.77); 207 (3.98) IR (KBr): 713, 973, 994, 1050 (sh), 1071 (sh), 1098, 1154, 1183, 1232 (sh), 1274, 1378, 1459, 1723, 2933, 2952, 3377, 3413, 3451 cm-1.

MS (FAB) (neg. Ions) = 508 (MH) - DC (DC-Aluminum foil 6OF254, Merck, propellant: dichloromethane / acetone 90/10) Rf = 0.3 (detection by spraying with anisaldehyde / sulfuric acid reagent and by heating to 120 ° C) HPLC (column: 4 x 25 0 mm Nucleosil 100-7 ° C, Macherey Nagel, flow 1.5 ml / min., propellant methanol / water 70/30 in 20 minutes linearly to 100/0 R f = 9.0 min.

The NMR values of both compounds are as follows.

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Table 1 1 H-NMR values (CDCl 3, δ, ppm)

SORAPHEN A_B

2 3.14 3.08 3 4 3.18 3.12 5 4.04 4.04 6 1.94 1.82 7 3.83 3.92 8 2.51 1.88 9 6.19 10 5, 48 1.80 / 1.402) 11 3.69 3.793) 12 3.43 3.353) 13A / B 1.69 / 1.26 1.59 / 1.352) 14A / B 1.37 / 1.17 15A / B 1, 48 16A / B 1.69 / 2.10 17 5.86 5.72 18 1.12 1.10 19 3.38 3.41 20 1.05 1.01 21 1.03 0.84 22 3.29 23 3.44 3.36 Γ - 2 '/ 6': 3 '/ 5' 7.27-7.38 7.25-7.36 4 '3-OH 4.43 4.68 5-OH 3, 56 1): multipets 1.25 to 1.8 ppm 2), 3): the order can be changed in each case

II

17 9301 4

Table 2 13 C-NMR values (CDCl 3, δ, ppm)

SORAPHEN A_B

1,170.8 171.6 2 46.3 45.7 3 99.5 99.5 4 76.3 76.4 5 68.9 69.3 ^ 6 35.6 35.1 7 42.4 71.3 8 35.3 32.5 9 139.6 27.92> 10 122.8 27.32)

11 85.0 68.91J

12 82.8 69.3 ° 13 30.4 25.42) 14 23.3 24.72) 15 25.7 22.82) 16 35.7 34.6 17 74.4 75.7 18 11.5 11.6 19 57.2 57.43) 20 10.2 10.2 21 12.4 14.1 22 56.1 23 58.0 57.23) 1 '141.1 140.7: 2'; 6 '126.2 126.4 3'; 5 '128.5 128.4 4' 128.0 128.0 1); 2); 3): The order can be changed at any time. Based on the X-ray structure analysis and the values clarified above, the following space structures Ia 'and Ib' are assumed for sorafen As lie (= compound Ia) and sorafen Bs (= compound Ib): «« ia 9301 4 2 2 gcHi / \ / \ / \ / " > CH 3 I 20 V * 3 ✓ \. 013 i «?! ./\/'Y'\4/ I II! 6chj Y / e">

HO

• · · XHj / \ / \ / \ / I Ach3! \ n n '\ / CH3 i f I i (lb ·) / VV \ 4V'S ° “I II I 6ch3 \ /

In particular, the invention also comprises the compounds Ia 'and Ib', their preparation and their use as plant microbicides.

The spectrum of activity of compounds Ia and Ib in the plate diffusion test

The concentration of the compounds is 1 μg / test leaf, agar volume 10 ml / plate (petri dishes, diameter 9 cm). The same values were determined for compound Ia and Ib.

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Test organism Barrier diameter [mm]

Debaryomyces hansenii 29

Candida albicans 25

Hansenula anomala 15

Nadsonia fulvescens

Nematospora coryli 35

Rhodotorula qlutinia 21

Saccaromyces cerevisiae 20

Schi2osaccharomyces pombe Torulopsis glabrata

Alternaria solani 27

Botrytis cinerea 30

Mucor hiemalis 30

Pythium debaryanum 45

Rhizopus arrhizus 10

The minimum inhibitory concentration MIC also gives the same values for both compounds Ia and Ib.

The minimal

Organismi_MNK / y q / mL

Nematospora coryli 0.05

Candida albicans 0.06

Rhodotorula gluten 1.0

Saccharomyces cerevisiae 2.0

Torulopsis glabrata 3.0

Nadsonia fulvescens 3.0

Mucor hiemalis 0.03 '·' The inhibitory effect on Candida is still reversible after 24 hours of exposure. Growth of Nematospora coryl ceases 90 minutes after the addition of Compound I. At the same time, DNA and protein synthesis are regulated.

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Serum Binding: Bovine serum has no effect on the efficacy of Compound I.

2. Preparation examples of the active substance of the formula I (% =% by weight) 2.1 Emulsion concentrates a) b) c) active substance 25% 40% 50%

Ca-dodecylbenzenesulfonate 5% 8% 6% castor oil polyethylene glycol ether (36 moles ethylene oxide) 5% tributylphenoyl polyethylene glycol ether (30 moles ethylene oxide) - 12% 4% cyclohexanone - 15% 20% 20% prepares by dilution with water emulsions of the desired concentration.

2.2 Solutions a) b) c) d) active substance 80% 10% 5% 95% ethylene glycol monomethyl ether 20% - - polyethylene glycol MG 400 - 70%: N-methyl-2-pyrrolidone - 20% epoxidized coconut oil - - 1% 5% petrol (boiling range 160-190 ° C) - - 94% (MG = molecular weight)

The solutions can be used in the form of very fine droplets.

»K.

21 9301 4 2.3 Granules a) b) active substance 5% 10% kaolin 94% highly dispersed silicic acid 1% attapulgite - 90%

The active ingredient is dissolved in methylene chloride, sprayed onto the carrier and the solvent is then evaporated in vacuo.

2.4 Dusts a) b) active substance 2% 5% highly dispersed silicic acid 1% 5% talc 97% kaolin - 90%

By effectively mixing the carriers with the active ingredient, ready-to-use dusts are obtained. These can be ground by adding more of these three carriers to ready-to-use dusts with an active ingredient content of 0.001%.

2.5 Spraying powders a) b) c) active substance 25% 50% 75%

Na-lignin sulfonate 5% 5%

Na-lauryl sulphate 3% to 5%

Na-diisobutylnaphthalenesulfonate - 6% 10% octylphenol polyethylene glycol ether - 2% - (7-8 moles of ethylene oxide) highly dispersed silicic acid 5% 10% 10% kaolin 62% 27% 22 9301 4

The active ingredient is mixed well with the additives and ground in a suitable mill. An injection powder is obtained which can be diluted with water to suspensions of the desired concentration.

2.6 Coated granulate active substance 3% polyethylene glycol (MG 200) 3% kaolin 94% (MG = molecular weight)

The finely ground active ingredient is spread evenly in a mixer on kaolin moistened with polyethylene glycol. In this way, dust-free coated granules are obtained.

2.7 Suspension concentrate of the active substance 40% ethylene glycol 10% nonylphenol polyethylene glycol (15 moles of ethylene oxide) 6%

Na-lignin sulfonate 10% carboxymethylcellulose 1% 37% aqueous formaldehyde 0.2% silicone oil in the form of a 75% aqueous emulsion 0.8% water 32%

The finely divided active substance is mixed thoroughly with the additives. This gives a suspension concentrate from which suspensions of the desired concentration can be prepared by diluting with water.

li 23 * 3ϋ14 2.8 Biomass concentrate

The biomass from the production culture is dried, ground and mixed with ethylene glycol monomethyl ether in a weight ratio of 80:20. Such a concentrate can be arbitrarily diluted with water to spray suspensions.

3. Biological examples (In the following, "active substance" means, without further mention, both "sorafen A" and "sorafen B")

Example 3.1: Action against Puccinia graminis in wheat a) Residual protection effect

Wheat seedlings were sprayed 6 days after sowing with a spray liquor prepared from the spray powder of the active substance (0.02% of active substance). 24 hours ku- ·. The treated plants were infected with a fungal uredospor suspension. Incubate for 48 hours at 95-100% relative humidity and about 20 ° C, after which the infected seedlings were placed in a greenhouse at about 22 ° C. Evaluation of rust spots was performed 12 days after infection.

b) Systemic impact

After 5 days of sowing, spraying broth (0.002% of active ingredient relative to the area) was sprayed on the wheat seedlings.

After 48 hours, the treated seedlings were infected with a fungal uredosporis suspension. Incubate for 48 hours at 95-100% relative humidity and about 20 ° C, after which the infected seedlings were placed in a greenhouse at about 22 ° C. Evaluation of rust spots was performed 12 days after infection.

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Fungal disease was completely prevented in both experiments with the active substance.

In contrast, untreated but infected control seedlings had 100% Puccinia disease.

Example 3.2: Action against Phytophthora in tomato or If a) Residual protective effect After 3 weeks of cultivation, the tomato seedlings were sprayed with a spray liquor prepared from a spray powder of the active substance (0.02% of active substance). After 24 hours, the treated seedlings were treated with a sporangiosus fungus of the fungus. The evaluation of the fungal disease took place after incubation of the seedlings for 5 days at 90-100% relative humidity and 20 ° C.

b) Systemic impact

After 3 weeks of cultivation, the tomato seedlings were poured into a spray broth (0.006% of active ingredient based on the amount of soil) of the active substance spray powder. Care was taken that the spray liquor did not come into contact with the above-ground parts of the seedlings.

After 48 hours, the treated seedlings were infected with a fungus; sporangiosuspensiolla. Infected seedlings were incubated for 5 days at 90-100% relative humidity and 20 ° C, after which the presence of fungal disease was assessed.

No fungal disease was detected in both experiments.

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Example 3.3: Action against Plasmopara viticola in vines a) Residual protection effect In a 4- to 5-leaf stage, the vine seedlings were sprayed with the active substance by spraying with a spray broth (0.006% of active substance). After 24 hours, the treated seedlings were infected with a sporangio suspension of the fungus. Incubate for 6 days at 95-100% relative humidity and 20 ° C, after which fungal disease was assessed.

Compared to untreated but infected control seedlings with 100% fungal disease, the seedlings treated with active ingredient I were free of fungal disease.

Example 3: 4; Action against Cercospora arachidicola in peanut seedlings Residual protective effect Peanut seedlings 10-15 cm high were sprayed with a spray of active substance in a spray broth (0.006% of active substance) and after 48 hours they were infected with a fungal conidia suspension. The infected seedlings were incubated for 72 hours at about 21 ° C and high humidity and then placed in a greenhouse until typical leaf spots appeared. Evaluation of the fungicidal effect was performed 12 days after infection based on the number and size of spots present.

The seedlings treated with active ingredient I were intact. There was also no disease in the seedlings treated with sorafen A at the end of the evaluation using a spray concentration of 0.002%. In contrast, untreated but infected control seedlings showed 100% Cercospora disease ♦

Example 3.5; Effect against Venturia inaequalis in apple plants Residual protection effect

Apple cuttings with new shoots 10 to 20 cm long were sprayed with a spray liquor made from the spray powder of the active substance (0.02% of active substance). After 24 hours, the treated seedlings were infected with a fungal conidia suspension. The seedlings were then incubated for 5 days at 90-100% relative humidity and placed in a greenhouse at 20-24 ° C for 10 days. Scabies was assessed 15 days after infection.

The cuttings treated with the active substance were disease-free.

Example 3.6; Effect against Botrytis cinerea in apples Residual protection effect

Artificially damaged apples were treated by instilling a spray broth (0.02% of active ingredient) prepared from the spray powder of the active substance into the damaged areas. The treated apples were then inoculated with a spore suspension of the fungus and incubated for one week at high humidity and about 20 ° C. In the evaluation, the contaminated damaged sites were calculated and the fungicidal effect of the test substance was deduced from this.

The active substance completely inhibited the growth of the fungus.

Il.

„9301 4

Example 3.7; Effect against Erysiphae qramin in barley a) Residual protective effect Approximately 8 cm high barley seedlings were sprayed with the active ingredient sprayed with powdered broth (0.006% active substance). After 3-4 hours, the treated seedlings were pollinated with fungal conidia. Infected barley seedlings were placed in a greenhouse at about 22 ° C and fungal disease was assessed after 10 days.

b) Systemic effect Spray broth (0.002% active substance relative to the ground) was sprayed on barley seedlings about 8 cm high. In doing so, care was taken to ensure that the spray broth did not come into contact with the above-ground trout parts. After 48 hours, the treated seedlings were pollinated with fungal conidia. Infected barley seedlings were placed in a greenhouse of approx.

. At 22 ° C and fungal disease was assessed after 10 days.

The seedlings were disease-free in both experiments, but all control seedlings showed disease.

Example 3.8: Action against Rhizoctonia solania (soil: fungus on rice plants)

Protective topical application to 12-day-old rice seedlings Spray broth (0.002% active substance) prepared from the active ingredient preparation was poured without contaminating the above-ground parts of the seedlings. To infect treated seedlings, a suspension of R. solan mycelium and mycelium warts is poured on the ground. Incubated for 6 days at 27 ° C (day) or resp. At 23 ° C (night) and 100% relative humidity (humidification box) in an air conditioning chamber, the fungal disease is assessed in the leaf sheath, leaves and stems.

After treatment with the active substance, no disease occurs. 1

II

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Claims (4)

29 9 3014 1. Sorafen of the formula I? Jf »! s / WW \ h k 3 'wherein R represents either methyl and has a double bond at the 9,10 position or wherein R represents hydrogen and has a single bond at the 9,10 position. Process for the preparation of sorafen according to Claim 1, characterized in that the microorganism Sorangium (Polyangium) cellulosum "So ce 26" (NCIB 12 411) is cultured in a culture medium which in each case contains at least one assimilable C-source and N-source and corresponding inorganic sources. salts, at 10-35 ° C in the presence or absence of an adsorber resin, then the culture broth or the filtered adsorbent resin, respectively, is extracted with a suitable solvent phase, the resulting solution is evaporated and the residual residue is purified, if necessary, by chromatography and / or recrystallization. The sorafen or active ingredient according to claim 1. The use of a substance containing at least one compound of the formula I as claimed in claim 1 in combination with a suitable carrier as a conventional substance for controlling plant diseases. Use according to Claim 3, characterized in that the essential component of the substance used is a biomass containing a compound of the formula I prepared by the process according to Claim 2.