WO2003044005A1 - Cruentaren a and b as pharmaceuticals and agrochemicals - Google Patents

Abstract

Compounds of the general formula (la) or (Ib) and pharmaceutically and agrochemically acceptable salts thereof and all geometric isomers and stereoisomers, and mixtures thereof in enriched form, are suitable for use as cytotoxic active ingredients, bactericides, fungicides, insecticides, acaricides and nematicides. Such metabolites of formulae (la) and (Ib) are obtainable by fermentation of a microorganism of the species Byssophaga cruenta, nov. comb., suborder Sorangineae, order Myxococcales.

Description

CRUENTAREN A AND B AS PHARMACEUTICALS AND AGROCHEMICALS

The present invention relates to novel, pharmaceutically and agrochemically active benzofused lactones of formulae la and lb hereinbelow, and especially to the specific isomers thereof designated cruentaren A and B, to a process for their preparation by growing/culturing cell cultures of the bacterium Byssophaga cruenta, nov. comb., strain Ha r1 , suborder Sorangineae, order Myxococcales, to compositions comprising such compounds, and to their biological activity profile and their pharmaceutical and agrochemical use, e.g. as cytotoxic compounds, or in the control of bacteria, yeasts, fungi, insects and acarids.

In 1897, R. Thaxter described the unique life cycle of myxobacteria in Bot. Gaz. 23, 395-411 (1897), specifically using the new species Myxococcus cruentus as an example. The organism, which had a characteristic bluish-red colour, was discovered in cow dung in the region of Burbank, Tennessee.

Since then, it has not been possible to find any further information about Myxococcus cruentus, and the genus has therefore been deleted from the list of names.

Surprisingly, a culturable organism having a conspicuously red pigmentation atypical of myxobacteria has now been discovered at the Gesellschaft fur Biotechnologische Forschung mbH (GBF) in Braunschweig, Germany.

The newly discovered myxobacteria strain, in addition to possessing other characteristics, has as a characteristic feature the ability to degrade cellulose. This is an indication of a new genus. The vegetative cells are typical of the Sorangineae suborder, and classification of the strain under the new genus Byssophaga, reusing the species name "cruenta" already used by Thaxter, is therefore proposed.

Other cellulose-degrading strains of myxobacteria have in the meantime been described by

Pronina in Microbiology 31 , 384-390 (1962).

The production strain designated Ha r1 of the bacterium Byssophaga cruenta, nov. comb., suborder Sorangineae, order Myxococcales, now in question was isolated in January 1997 by GBF from a soil sample from the sage (salvia) plain south of Holbrook, Arizona, U.S.A.. The organism can be grown successfully by standard methods by applying small specimen pieces to living Escherichia cultures in aqueous agar. Very good growth of the organism can also be achieved on agar with intact cells of baker's yeast, or on filter paper laid on VY/2 agar, or on MM1 agar in which cellulose powder has been suspended. The filter paper or the cellulose powder is completely digested in the course of the process. Finally, good growth can also be observed on chitin agar with degradation of the chitin when the uppermost chitin layer is on aqueous agar (Chit7-agar). Cycloheximide was added to the agar in order to suppress fungal growth (H. Reichenbach and M. Dworkin, "The Myxobacteria", in A. Balows, H. G. Trϋper, M. Dworkin, W. Harder and K. H. Schleifer (Editors) "The Prokaryotes", 2^nd Edition, pages 3416-3487, Springer Verlag, New York, 1992).

The new organism, which has the ability to produce the pharmaceutically and agrochemically active isomers cruentaren A and B, grows extremely well on living E. coli by decomposing those nutrient bacteria. It is thus unlike Sorangium, which neither grows on E. coli bacteria nor is able to decompose them.

The present invention thus relates to the compound of the general formula la or lb

or

and to the pharmaceutically and agrochemically acceptable salts thereof and all geometric isomers and stereoisomers thereof, and to a mixture thereof in enriched form.

The compounds of formulae la and lb having an acid hydrogen, for example in the phenol group, can also be in salt form. The compounds of formulae la and lb can also optionally be isolated and produced in pure form by salt formation.

Suitable pharmaceutically and agrochemically acceptable salts with bases include, for example, metal salts, such as alkali metal and alkaline earth metal salts, e.g. sodium, potassium or magnesium salts, or salts with ammonia or an organic amine, such as morpholine, piperidine, pyrrolidine, a mono-, di- or tri-lower alkylamine, e.g. ethyl-, diethyl-, triethyl- or dimethyl-propylamine, or a mono-, di- or tri-hydroxy-lower alkylamine, e.g. mono-, di- or tri-ethanolamine. The free form, that is to say the neutral form, is preferred. Hereinbefore and hereinafter, any reference to the free compounds of formulae la and/or lb and their salts is to be understood as including also the corresponding pharmaceutically and agrochemically acceptable salts and the free compounds of formulae la and/or lb, respectively, as appropriate.

Salts of compounds of formulae la and lb can be prepared in a manner known per se. For example, salts with bases are obtained by treatment with a suitable base or with a suitable ion exchange reagent.

Salts with bases of compounds of formulae la and lb can be converted into the free compounds of formulae la and lb in customary manner, for example by treatment with a suitable acid or with a suitable ion exchange reagent.

Salts of compounds of formulae la and lb can be converted into different salts of compounds of formulae la and lb in a manner known perse.

In the process described hereinbelow, by fermentation of the myxobacterial strain designated Ha r1 the compounds of formulae la and lb are obtainable in the form of a mixture, which can be enriched by customary separation methods, for example chromatographic methods, separated on the basis of the physicochemical properties, for example by means of high pressure liquid chromatography (HPLC), into the pure specific isomers, and also separated off from structural isomers and other myxobacteria metabolites. In the present invention, the compounds of formulae la and lb are to be understood as meaning both the racemates (having (R)- and (S)-configured 8 stereogenic centres C-9, C-10, C-15, C-16, C-17, C-18, C-24 and C-25) and all of the possible 256 optical isomers (having (R)- or (S)-configured 8 stereogenic centres C-9, C-10, C-15, C-16, C-17, C-18, C-24 and C-25) in enriched form or in the form of pure optical isomers, but especially in the form of the pure optical isomers that can be isolated from the fermentation process described for the production strain Ha r1.

By virtue of the presence of alicyclic and aliphatic double bonds in the compounds of formulae la and lb, geometric isomerism (E/Z) may also occur.

Formulae la and lb are to include all such possible isomers, enantiomers and diastereo- isomers, and also mixtures thereof.

Preference is given to the corresponding specific isomer of the compound of the general formula la or lb designated cruentaren A (formula la) or cruentaren B (formula lb), which is obtainable by fermentation of a bacterial strain of the species Byssophaga cruenta, nov. comb., suborder Sorangineae, order Myxococcales. An example of that species designated Ha r1 was deposited by GBF - Gesellschaft fϋr Biotechnologische Forschung mbH, Mascheroder Weg 1 , 38124 Braunschweig on 26th September 2001 under deposit No. DSM 14567 at the Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) in Braunschweig, Germany.

Of those, special preference is given to cruentaren A.

Preference is likewise given to an enriched mixture containing from 80 to 99% by weight of the specific isomers of the compounds of the general formulae la and lb designated cruentaren A and cruentaren B, which mixture is obtainable from fermentation of the production strain Ha r1 of the myxobacterium Byssophaga cruenta.

The present invention relates also to: a) the present microorganism of the order Myxococcales, b) the preparation of compounds of the general formulae la and lb and of the two metabolites cruentaren A and B in the form of an enriched mixture or in the form of isomerically pure individual components by the growth/culturing/fermentation of cell cultures of the bacterium Byssophaga cruenta, production strain Ha r1 , and, where appropriate, isomerisation or racemisation of certain stereogenic centres of cruentaren A and B (see above), and also c) their pharmaceutical, and d) agrochemical use, e.g. as cytotoxic active ingredients, as bactericides, fungicides, insecticides and acaricides.

Description of the production strain Ha r1 :

The vegetative cells of Ha r1 are cylindrical with broadly rounded-off ends, 0.9-1.2 x 4-6 μm. On solid nutrients, the organism grows well on yeast agar, for example VY/2 agar (baker's yeast, 0.5% based on fresh weight; calcium chloride • 2 H₂O 0.1 %; cyanocobolamin 0.5 mg/I; agar 1.5%; pH 7.2). When small pieces of filter paper are arranged on the VY/2 plates, or maltose is added to the agar, increased cell growth is observed with degradation of the cellulose. In contrast, no cell growth is observed on pure glucose/mineral salt media. The strain grows aerobically preferably at a temperature of 30°C in the neutral pH range. On suitable nutrient media, the cell culture gradually spreads out over the culture plate. When yeast cells are present in the nutrient medium, they are substantially degraded. Chitin, too, is degraded by the strain in question.

In liquid media, the strain grows into small red cell-clusters, both in shake flasks at 160 revs/min (revolutions per minute) (100 ml of medium in a 250 ml Erlenmeyer flask or 400 ml of medium in a 1000 ml Erlenmeyer flask) and in bioreactors (tested up to a size of 100 litres). A suitable culture medium is, for example, VY/2 medium (consisting of baker's yeast 0.5% based on fresh weight; calcium chloride • 2 H₂O 0.1%; cyanocobolamin 0.5 mg/I; pH 7.2) with the addition of 0.2-0.5% maltose and 50 mM 4-(2-hydroxyethyl)-piperazine-1 - ethanesulfonic acid (= HEPES as biological buffer).

The cultures are maintained at 30°C for from 5 to 7 days.

The production strain Ha r1 can be preserved for several years, for example by freezing the vegetative cells of agar plates or liquid cultures in VY/2 medium at a temperature of -80°C or in liquid nitrogen. The compounds of the general formulae la and lb, or cruentaren A and B in the form of an enriched mixture, or cruentaren A or cruentaren B as individual components, are obtained by the growth/culturing/fermentation of the bacterium Byssophaga cruenta, deposited under deposit number DSM 14567, in a nutrient medium (bioreactor) with carbon and nitrogen sources, as well as mineral salts, by removal of the cell mass obtained (co-fermentation with adsorber resin, e.g. Amberlite XAD 16) and subsequent extraction with acetone, by concentration of the combined acetone phases up to the aqueous phase and extraction thereof with ethyl acetate, by drying of the organic phase over a drying agent, removal of the solvent at reduced pressure and, first, for the preparation of the enriched mixture containing 80-99% of the compounds of the general formulae la and lb or cruentaren A and B, allowing the resulting crude concentrate to run through an open silica gel column using a sequence of gradient eluants consisting of ethyl acetate/n-heptane, ethyl acetate, and ethyl acetate/methanol and then, for the preparation of the individual components cruentaren A and B in pure form, allowing that enriched mixture to run through a preparative RP-18 column ("reversed phase") using acetonitrile/water as eluant and, where appropriate, converting cruentaren A and B into their pharmaceutically and agrochemically acceptable salts.

The absolute configuration of the resulting specific isomers of the compounds of the general formulae la and lb designated cruentaren A and B is awaiting clarification.

The quantitative determination of cruentaren in the extracts is carried out by HPLC, and the working up, isolation and chemical characterisation are carried out as described hereinbelow.

A. Fermentation Example A1. Production strain: Ha r1 of the bacterium Byssophaga cruenta, nov. comb., suborder Sorangineae, order Myxococcales.

A2. Origin of the production strain: The strain designated Ha r1 was isolated at GBF from a soil sample collected south of Holbrook, Arizona, U.S.A. and deposited by GBF at the Deutschen Sammlung von Mikroorganismen und Zellkulturen (DSMZ) in Braunschweig, Germany, under deposit number DSM 14567. A3. Fermentation Example in a bioreactor:

A bioreactor from "Bioingeneering" with a total volume of 150 litres and a working volume of 90 litres is equipped with a paddle stirrer. The nutrient medium is VY/2 + 0.3% maltose + 10 mM HEPES (= 4-(2-hydroxyethyl)-piperazine-1-ethanesulfonic acid), with 1% adsorber resin, Amberlite XAD 16, additionally being added to the nutrient medium. 85 litres of the nutrient medium are inoculated with 5 litres of culture taken from shake flasks exhibiting good growth.

At the start of the fermentation, the aeration rate is set at 0.05 wm (= aeration volume per minute with respect to the total volume of the fermenter, that is to say 1/20 of the fermenter volume per minute) and the speed of the stirrer is set at 100 revs/min.. The pO₂ value (= oxygen pressure), which at the start of fermentation is 98% saturation, falls towards the end of fermentation and after 14 days is down to 78%. The pH value for the entire duration of the fermentation ranges from 7.0 to 7.2. The content of pure cruentaren A at the end of fermentation is 2.7 mg/I (HPLC evidence).

B. Isolation and physicochemical characterisation of cruentaren A and B

B1. Isolation of cruentaren A and B

After co-fermentation with 0.9 litre of the adsorber resin Amberlite XAD 16, the said resin is removed from the fermentation liquor. The adsorber resin is then extracted in 4 batches in each case with 3 litres of acetone, the combined acetone extracts are concentrated in vacuo (200 mbar) at 30°C up to the aqueous phase, and that phase is extracted three times with 500 ml of ethyl acetate each time. The combined organic phases are dried over magnesium sulfate, filtered and concentrated by evaporation at 150 mbar and 30°C. The residue obtained is 10.0 g of crude extract, which is chromatographed on an open silica gel glass column (diameter: 6 cm, height: 14 cm) filled with 180 g of silica gel 60 (Merck,

15-40 μm). The eluant used is a gradient eluant with, first, ethyl acetate in n-heptane, then ethyl acetate, and finally ethyl acetate in methanol, the composition being as follows:

1 st 800 ml of ethyl acetate/n-heptane 2/1 ;

2nd 300 ml of ethyl acetate;

3rd 700 ml of ethyl acetate/methanol 4/1 ; and

4th 800 ml of ethyl acetate/methanol 3/2.

After from 150 to 180 minutes, the two cruentarens A and B, in the form of an enriched mixture, are eluted with the 3rd eluant at a flow rate of 10 ml/minute, detection being carried out by means of thin-layer chromatography. After concentration of the mixed fractions containing cruentaren by evaporation in vacuo, a residue of 0.32 g is obtained.

The further separation of the enriched mixture into the individual components cruentaren A and B is carried out on a preparative RP-18 column (Kronlab ODS AQ 120 A, 16 μm, 48 x 3 cm) with the eluant acetonitrile/water 55/45 and at a flow rate of 18 ml/minute and detection by UV absorption at 206 nm.

Under those conditions, cruentaren A is eluted after 75-85 minutes and cruentaren B after 115-125 minutes. The two separated fractions containing cruentaren A and B are concentrated in vacuo up to the aqueous phase and extracted twice with 100 ml of ethyl acetate each time. The combined ethyl acetate extracts are concentrated. After drying in vacuo, 180 mg of cruentaren A and 9 mg of cruentaren B are obtained.

B2. Physical data of cruentaren A:

Empirical formula: C₃₃H₅₁NO₈

Molecular weight: 589

Mass spectrum: MAT 95 DCI (M + H)⁺ found: 590.37052; calculated: 590.36929.

UV (methanol) λ_max [nm] (logε): 216 (4.26); 265 (3.98); 302 (3.66); 346 (1.94).

IR (KBr) [cm^"1]: 3396, 2959, 2932, 2863, 1646, 1614, 1251 , 1159.

Optical rotation: α_{D =} 3.4 ° (c = 13.3 mg/ml in methanol).

Thin-layer chromatography: eluant: ethyl acetate; R_f = 0.53.

Identification by means of analytical HPLC (column: Macherey Nagel 125/2 120 5 μm C-18; eluant: acetonitrile/water 55/45; flow rate: 0.3 ml/minute; detection: diode array):

R_t 3.9 minutes.

Table 1 : ¹H- and ¹³C-NMR spectra of cruentaren A in CDCI₃ (600 and 100 MHz)

B3. Physical data of cruentaren B

Empirical formula: C₃₃H₅₁NO₈

Molecular weight: 589

Mass spectrum: MAT 95 DCI (M + H)⁺ found: 590.37052; calculated: 590.36929.

UV (methanol) λ_max [nm] (logε): 210 (4.36); 215 (4.38); 267 (4.14); 301 (3.77).

IR (KBr) [cm^"1]: 3416, 2963, 2930, 2869, 1661 , 1628, 1250, 1159.

Optical rotation: α_D = 9.1 ° (c = 6.6 mg/ml in methanol).

Thin-layer chromatography: eluant: ethyl acetate; R_f = 0.40.

Identification by means of analytical HPLC (column: Macherey Nagel 1 5/2 120 5 μm C-18; eluant: acetonitrile/water 55/45; flow rate: 0.3 ml/minute; detection: diode array):

R_t 5.0 minutes

Table 2: ¹H- and ¹³C-NMR spectra of cruentaren B in CDCI₃ (600 and 100 MHz)

Bioloqical activity profile of the production strain Ha r1 :

In the cell extracts of Ha r1 it is possible on the one hand to detect a cytotoxic activity with respect to various cell cultures, e.g. L929 mouse fibroblasts, and on the other hand good activity against yeasts and fungi, and somewhat weaker activity against bacteria (agar diffusion test).

The substances responsible for those activiities are cruentaren B and, especially, cruentaren A.

For qualitative determination of cruentaren, the cell mass of the bacterium Byssophaga cruenta is extracted with a suitable organic solvent, for example acetone, or the fermentation culture is extracted by stirring, after the addition of adsorber resin Amberlite XAD 16 (Rohm & Haas), and then the separated XAD is eluted with acetone or methanol, and aliquots of the concentrated extracts are tested against L929 mouse fibroblasts or on Saccharomyces cerevisiae agar plates.

The compounds of formulae la and lb and their specific isomers cruentaren A and B can be used in unmodified form, that is to say as obtained from the fermentation culture, in human and veterinary medicine and in agriculture, but they are preferably processed in conventional manner with the adjuvants customary in formulation technology.

The present invention accordingly relates also to a pharmaceutical composition consisting of a compound of formula la and/or lb or the corresponding specific isomer cruentaren A and/or B together with the adjuvants customary in pharmaceutical formulation technology, for example solid carriers or solvents.

The compositions according to the invention can be prepared in a manner known perse, for example by means of conventional mixing, granulating, confectioning, dissolving or lyophilising processes. For example, pharmaceutical compositions for oral administration can be obtained by combining the active ingredients with solid carriers, optionally granulating a resulting mixture, and processing the mixture or granules, if desired or necessary, after the addition of suitable adjuvants, to form tablets or dragee cores.

Examples of such formulation adjuvants are given, for example, in EP-A-0 564409, US-A-6 271 222 and US-A-6 284 753. The present compositions can be used in pharmaceutical treatment, e.g. as cytotoxic agents or in the control of pathogenic organisms (bacteria, fungi) in and on humans and animals.

The present invention relates also to an agrochemical composition consisting of compounds of formula la and/or lb or the appropriate specific isomer(s) cruentaren A and/or B together with adjuvants customary in agrochemical formulation technology.

C. Biological characterisation and activity of cruentaren

C1. Antimicrobial activity of cruentaren A:

Surprisingly, it has now been found that cruentaren A possesses antibiotic activity, which is ascertained by means of the agar diffusion test by reference to the diameter of the inhibiting areola, or from the culture density in the dilution series test. For example, cruentaren A applied in the form of a methanolic stock solution inhibits the growth of some fungi and yeasts. The maximum inhibitory concentration (MIC) in the case of Saccharomyces cerevisiae (N3 medium: bacto-peptone 1 %, bacto-yeast extract 1 %, glycerol 2%, dissolved in 50 ml sodium/potassium phosphate buffer pH 6.3, agar 2%) is 0.4 μg/ml.

Cruentaren A is effective at 20 μg/test platelet against the following test organisms listed in

Table 3.

Table 3: Diameter of the inhibiting areola of test organisms on the application per platelet of 20 μg of cruentaren A dissolved in methanol.

In Table 3, Myc(= mycophil) medium: phytone-peptone 1%, glucose 1 %, agar 1.6%.

Cruentaren B does not exhibit any antimicrobial activity

C2. Cytotoxic activity of cruentaren A:

A further surprising property of cruentaren A is the very appreciable cytotoxic activity in a variety of human and animal cell lines, which are completely inhibited, e.g. the mouse L929 cell culture.

Resistant cell cultures (PC-3) are also found.

Typical morphological changes to cells are not observed.

The following Table 4 lists the concentrations of cruentaren A dissolved in methanol for a

50% inhibition (IC₅₀) in growth of various cell cultures.

Table 4: 50% inhibition (IC₅₀) of various human and animal cell cultures by cruentaren A in ng/ml in methanol.

Compared with cruentaren A, cruentaren B is cytotoxically less active, for example an IC₅₀ inhibition of L929 mouse cells is observed at 1000 ng/ml.

C3. Agrochemical activity of cruentaren A and B:

A further special property of cruentaren A and/or B is the surprisingly broad activity in relation to phytopathogenic insects, acarids and nematodes, and in relation to animal parasites, especially parasites in mammals, e.g. against worms, nematodes, cistodes and trematodes. The novel compounds can therefore be used successfully in agriculture against all development stages of parasites, especially insects, acarids and nematodes, that occur on cultivated plants, for example cereals, maize, cotton, soybean, sugar beet, sugar cane, plantation crops, rape, rice, vegetables, fruits or ornamentals, or live in stored goods, and also that play a part in the hygiene sector or as parasites in and on domestic animals and productive livestock.

The novel compounds according to the present invention can be used successfully in the control of the following harmful organisms: of the order Lepidoptera. e.g.: Acleris spp., Adoxophyes spp., Aegeria spp., Agrotis spp., Alabama argillaceae, Amylois spp., Anticarsia gemmatalis, Archips spp., Argyrotaenia spp., Autographa spp., Busseola fusca, Cadra cautella, Carposina nipponensis, Chilo spp., Choristoneura spp., Clysia ambiguella, Cnaphalocrocis spp., Cnephasia spp., Cochylis spp., Coleophora spp., Crocidolomia binotalis, Cryptophlebia leucotreta, Cydia spp., Diatraea spp., Diparopsis castanea, Earias spp., Ephestia spp., Eucosma spp., Eupoecilia ambiguella, Euproctis spp., Euxoa spp., Grapholita spp., Hedya nubiferana, Heliothis (Helicoverpa) spp., Hellula undalis, Hyphantria cunea, Keiferia lycopersicella, Leucoptera scitella, Lithocollethis spp., Lobesia botrana, Lymantria spp., Lyonetia spp., Malacosoma spp., Mamestra brassicae, Manduca sexta, Operophtera spp., Ostrinia nubilalis, Pammene spp., Pandemis spp., Panolis flammea, Pectinophora gossypiella, Phthorimaea operculella, Pieris rapae, Pieris spp., Plutella xylostella, Prays spp., Scirpophaga spp., Sesamia spp., Sparganothis spp., Spodoptera spp., Synanthedon spp., Thaumetopoea spp., Tortrix spp., Trichoplusia ni and Yponomeuta spp.; of the order Coleoptera, e.g.: Agriotes spp., Anthonomus spp., Atomaria linearis, Chaetocnema tibialis, Cosmopolites spp., Curculio spp., Dermestes spp., Diabrotica spp., Epilachna spp., Eremnus spp., Leptinotarsa decemlineata, Lissorhoptrus spp., Melolontha spp., Orycaephilus spp., Otiorhynchus spp., Phlyctinus spp., Popillia spp., Psylliodes spp., Rhizopertha spp., Scarabeidae, Sitophilus spp., Sitotroga spp., Tenebrio spp., Tribolium spp. and Trogoderma spp.; of the order Orthoptera. e.g.: Blatta spp., Blattella spp., Gryllotalpa spp., Leucophaea maderae, Locusta spp., Periplaneta spp. and Schistocerca spp.; of the order Isoptera, e.g.: Reticulitermes spp.; of the order Psocoptera. e.g.: Liposcelis spp.; of the order Anoplura, e.g.: Haematopinus spp., Linognathus spp., Pediculus spp.,

Pemphigus spp. and Phylloxera spp.; of the order Mallophaga. e.g.: Damalinia spp., Trichodectes spp. and Bovicola spp.; of the order Thvsanoptera e.g.: Frankliniella spp., Hercinothrips spp., Taeniothrips spp.,

Thrips palmi, Thrips tabaci and Scirtothrips aurantii; of the order Heteroptera, e.g.: Cimex spp., Distantiella theobroma, Dysdercus spp.,

Euchistus spp., Eurygaster spp., Leptocorisa spp., Nezara spp., Piesma spp., Rhodnius spp., Sahlbergella singularis, Scotinophara spp. and Triatoma spp.; of the order Homoptera, e.g.: Aleurothrixus floccosus, Aleyrodes brassicae, Aonidiella spp.,

Aphididae, Aphis spp., Aspidiotus spp., Bemisia tabaci, Ceroplaster spp., Chrysomphalus aonidium, Chrysomphalus dictyospermi, Coccus hesperidum, Empoasca spp., Eriosoma larigerum, Erythroneura spp., Gascardia spp., Laodelphax spp., Lecanium corni,

Lepidosaphes spp., Macrosiphus spp., Myzus spp., Nephotettix spp., Nilaparvata spp.,

Paratoria spp., Pemphigus spp., Planococcus spp., Pseudaulacaspis spp., Pseudococcus spp., Psylla spp., Pulvinaria aethiopica, Quadraspidiotus spp., Rhopalosiphum spp.,

Saissetia spp., Scaphoideus spp., Schizaphis spp., Sitobion spp., Trialeurodes vaporariorum, Trioza erytreae and Unaspis citri; of the order Hymenoptera, e.g.: Acromyrmex, Atta spp., Cephus spp., Diprion spp.,

Diprionidae, Gilpinia polytoma, Hoplocampa spp., Lasius spp., monomorium pharaonis,

Neodiprion spp., Solenopsis spp. and Vespa spp.; of the order Diptera, e.g.: Aedes spp., Antherigona soccata, Bibio hortulanus, Calliphora erythrocephala, Ceratitis spp., Chrysomyia spp., Culex spp., Cuterebra spp., Dacus spp.,

Dermatobia spp., Drosophila melanogaster, Fannia spp., Gastrophilus spp., Glossina spp.,

Haematobia spp., Hypoderma spp., Hyppobosca spp., Liriomyza spp., Lucilia spp.,

Melanagromyza spp., Musca spp., Oestrus spp., Orseolia spp., Oscinella frit, Pegomyia hyoscyami, Phorbia spp., Rhagoletis pomonella, Sciara spp., Stomoxys spp., Tabanus spp.,

Tannia spp. and Tipula spp.; of the order Siphonaptera, e.g.: Ceratophyllus spp., Xenopsylla cheopis, Ctenocephalides felis, Pulex spp. and Ctenocephalides canis; of the order Thvsanura, e.g.: Lepisma saccharina; and: of the order acarids. e.g.: Acarus siro, Aceria sheldoni, Aculus schlechtendali, Amblyomma spp., Argas spp., Boophilus spp., Brevipalpus spp., Bryobia praetiosa, Calipitrimerus spp.,

Chorioptes spp., Dermanyssus gallinae, Dermatophagoides spp., Dermamacentor spp.,

Eotetranychus carpini, Eriophyes spp., Haemaphysalis spp., Hyalomma spp., Ixodes spp., Myobia spp., Myocoptes spp., Olygonychus pratensis, Omithodoros spp., Panonychus spp., Phyllocoptruta oleivora, Polyphagotarsonemus latus, Psorergates spp., Psoroptes spp., Rhipicephalus spp., Rhizoglyphus spp., Sarcoptes spp., Tarsonemus spp. and Tetranychus spp.; of the class Nematoda. for example the families Filariidae and Setariidae, and the genera Haemonchus, Trichostrongylus, Ostertagia, Nematodirus, Cooperia, Ascaris, Bunostumum, Oesophagostonum, Chabertia, Trichuris, Strongylus, Trichonema, Dictyocaulus, Capillaria, Strongyloides, Heterakis, Toxocara, Oxyuris, Ancylostoma, Uncinaria, Toxascaris and Parascaris; and also Dirofilaria.

The present method of controlling nematodes is suitable also especially for soil-borne nematodes, because it is very efficient and environmentally friendly and is very well tolerated by useful plants. Such soil-borne nematodes that parasiticise on and in useful plants and their roots are, for example, from the genera Heterodera and Globodera (cystogenic nematodes), Meloidogyne (root-knot nematodes), Radopholus, Pratylenchus, Tylenchulus, Longidorus, Trichodorus, Xiphinema, Ditylenchus (stem parasites), Aphelenchoides (leaf nematodes) and Anguina (blossom nematodes).

An especially important aspect of the present invention is the use of the compounds of formulae la and lb according to the invention or the corresponding specific isomers cruentaren A and B in the protection of plants against parasitic feeding pests.

The compounds according to the invention can be used to control, that is to say to inhibit or destroy, pests of the type mentioned that occur on plants, especially on useful plants and ornamentals in agriculture, horticulture and forestry, or on parts of such plants, such as the fruit, blossom, leaves, stems, tubers or roots, while in some cases plant parts that grow later are still protected against those pests.

Target crops include especially cereals, such as wheat, barley, rye, oats, rice, maize and sorghum; beet, such as sugar beet and fodder beet; fruit, for example pomes, stone fruit and soft fruit, such as apples, pears, plums, peaches, almonds, cherries and berries, e.g. strawberries, raspberries and blackberries; leguminous plants, such as beans, lentils, peas and soybeans; oil plants, such as rape, mustard, poppy, olives, sunflowers, coconut, castor oil plants, cocoa beans and groundnuts; cucurbitaceae, such as marrows, cucumbers and melons; fibre plants, such as cotton, flax, hemp and jute; citrus fruit, such as oranges, lemons, grapefruit and mandarins; vegetables, such as spinach, lettuce, asparagus, cabbages, carrots, onions, tomatoes, potatoes and paprika; lauraceae, such as avocados, cinnamon and camphor; and also tobacco, nuts, coffee, aubergines, sugar cane, tea, pepper, vines, hops, bananas, natural rubber plants and ornamentals.

The active ingredients according to the invention are suitable especially for controlling Nila- parvata lugens, Frankliniella occidentalis, Spodoptera littoralis, Diabrotica balteata, Panonychus ulmi and Tetranychus urticae.

Further areas of use of the compounds according to the invention are the protection of stored goods and storerooms and the protection of raw materials, and also in the hygiene sector, especially the protection of domestic animals and productive livestock against pests of the type mentioned.

The invention accordingly relates also to pesticidal compositions, such as emulsifiable concentrates, suspension concentrates, directly sprayable or dilutable solutions, coatable pastes, dilute emulsions, wettable powders, soluble powders, dispersible powders, wettable powders, dusts, granules or encapsulations in polymeric substances, which comprise at least one of the compounds according to the invention, the type of formulation being chosen in accordance with the intended objectives and the prevailing circumstances.

The active ingredient is used in those compositions in pure form or, preferably, together with at least one of the adjuvants customary in formulation technology, such as extenders, e.g. solvents or solid carriers, or surface-active compounds (surfactants).

In plant protection, solvents, for example, may come into consideration, such as: aromatic hydrocarbons, which may be partially hydrogenated, preferably the C₈ to C₁₂ fractions of alkylbenzenes, such as xylene mixtures, alkylated naphthalenes or tetrahydronaphthalene, aliphatic or cycloaliphatic hydrocarbons, such as paraffins or cyclohexane, alcohols, such as ethanol, propanol or butanol, glycols and ethers and esters thereof, such as propylene glycol, dipropylene glycol ether, ethylene glycol or ethylene glycol monomethyl or monoethyl ether, ketones, such as cyclohexanone, isophorone or diacetone alcohol, strongly polar solvents, such as N-methylpyrrolid-2-one, dimethyl sulf oxide or N,N-dimethylformamide, water, vegetable oils, such as rapeseed oil, castor oil, coconut oil or soybean oil, which may or may not be epoxidised, and silicone oils. The solid carriers used e.g. for dusts and dispersible powders are normally natural mineral fillers, such as calcite, talcum, kaolin, montmorillonite or attapulgite. In order to improve the physical properties it is also possible to add highly dispersed silicic acids or highly dispersed absorbent polymers. Suitable granulated adsorptive carriers are porous types, for example pumice, broken brick, sepiolite or bentonite; and suitable non-sorbent carriers are calcite or sand. In addition, a great number of granulated materials of inorganic or organic nature can be used, especially dolomite or pulverised plant residues.

Depending on the nature of the active ingredient to be formulated, suitable surface-active compounds are non-ionic, cationic and/or anionic surfactants or surfactant mixtures having good emulsifying, dispersing and wetting properties. The surfactants listed hereinbelow are to be regarded only as examples; a large number of other surfactants customary in formulation technology and suitable in accordance with the invention are described in the relevant literature.

Suitable non-ionic surfactants are especially polyglycol ether derivatives of aliphatic or cycloaliphatic alcohols, saturated or unsaturated fatty acids and alkylphenols, said derivatives containing from 3 to 30 glycol ether groups and from 8 to 20 carbon atoms in the (aliphatic) hydrocarbon moiety and from 6 to 18 carbon atoms in the alkyl moiety of the alkylphenols. Also suitable are water-soluble adducts of polyethylene oxide with polyropylene glycol, ethylenediaminopolypropylene glycol and alkylpolypropylene glycol having from 1 to 10 carbon atoms in the alkyl chain, which adducts contain from 20 to 250 ethylene glycol ether groups and from 10 to 100 propylene glycol ether groups. These compounds usually contain from 1 to 5 ethylene glycol units per propylene glycol unit. Examples that may be mentioned are nonylphenol polyethoxyethanols, castor oil polyglycol ethers, polypropylene/polyethylene oxide adducts, tributylphenoxypolyethoxyethanol, polyethylene glycol and octylphenoxypolyethoxyethanol. Fatty acid esters of polyoxyethylene sorbitan, such as polyoxyethylene sorbitan trioleate, are also suitable.

Cationic surfactants are especially quaternary ammonium salts that contain as substituent at least one alkyl radical having from 8 to 22 C atoms and, as further substituents, unsubstituted or halogenated lower alkyl, benzyl or hydroxy-lower alkyl radicals. The salts are preferably in the form of halides, methyl sulfates or ethyl sulfates. Examples are stearyl- trimethyl-ammonium chloride and benzyl-di(2-chloroethyl)ethylammonium bromide. Both water-soluble soaps and water-soluble synthetic surface-active compounds are suitable anionic surfactants. Suitable soaps include the alkali metal salts, alkaline earth metal salts and unsubstituted or substituted ammonium salts of higher fatty acids (C₁₀ to C₂₂), such as the sodium or potassium salts of oleic or stearic acid, or of natural fatty acid mixtures, which can be obtained, for example, from coconut oil or tall oil; mention may also be made of fatty acid methyl taurine salts. More frequently, however, synthetic surfactants are used, especially fatty sulfonates, fatty sulfates, sulfonated benzimidazole derivatives or alkylarylsulfonates. The fatty sulfonates and fatty sulfates are usually in the form of alkali metal salts, alkaline earth metal salts or unsubstituted or substituted ammonium salts and generally contain an alkyl radical having from 8 to 22 C atoms, alkyl also including the alkyl moiety of acyl radicals; examples that may be mentioned are the sodium or calcium salts of lignosulfonic acid, of dodecylsulfate or of a fatty alcohol sulfate mixture prepared from natural fatty acids. These compounds also include the salts of the sulfuric acid esters and sulfonic acids of fatty alcohol/ethylene oxide adducts. The sulfonated benzimidazole derivatives preferably contain two sulfonic acid groups and one fatty acid radical having approximately from 8 to 22 C atoms. Examples of alkylarylsulfonates include, for example, the sodium, calcium or triethanolamine salts of dodecylbenzenesulfonic acid, of dibutylnaphthalenesulfonic acid or of a condensate of naphthalenesulfonic acid and formaldehyde. Also suitable are corresponding phosphates, such as salts of the phosphoric acid ester of an adduct of p-nonylphenol with 4 to 14 mol of ethylene oxide, or phospholipids.

The compositions for use in plant protection and in humans, domestic animals, productive livestock and pets, usually contain from 0.1 to 99%, especially from 0.1 to 95%, of active ingredient and from 1 to 99.9%, especially from 5 to 99.9%, of at least one solid or liquid adjuvant, it being possible generally for from 0 to 25%, especially from 0.1 to 20%, of the compositions to be surfactants (% in each case meaning per cent by weight). Whereas commercial products are preferably formulated as concentrates, the end user will normally employ dilute formulations that contain appreciably lower concentrations of active ingredient.

Preferred crop protection products have especially the following compositions (% = percent by weight):

Emulsifiable concentrates: active ingredient: 1 to 90%, preferably 5 to 20% surfactant: 1 to 30%, preferably 10 to 20% solvent: 5 to 98%, preferably 70 to 85%

Dusts: active ingredient: 0.1 to 10%, preferably 0.1 to 1 % solid carrier: 99.9 to 90%, preferably 99.9 to 99%

Suspension concentrates: active ingredient: 5 to 75%, preferably 10 to 50% water: 94 to 24%, preferably 88 to 30% surfactant: 1 to 40%, preferably 2 to 30%

Wettable powders: active ingredient: 0.5 to 90%, preferably 1 to 80% surfactant: 0.5 to 20%, preferably 1 to 15% solid carrier: 5 to 99%, preferably 15 to 98%

Granules: active ingredient: 0.5 to 30%, preferably 3 to 15% solid carrier: 99.5 to 70%, preferably 97 to 85%

The activity of the crop protection compositions according to the invention can be substantially broadened and adapted to prevailing circumstances by the addition of other insecticidal active ingredients. Examples of suitable additional active ingredients include, for example, representatives from the following classes of active ingredient: organophosphorus compounds, nitrophenols and derivatives, formamidines, acylureas, carbamates, pyrethroids, nitroenamines and derivatives, pyrroles, thioureas and derivatives, chlorinated hydrocarbons and Bacillus thuringiensis preparations. The compositions according to the invention may also comprise other solid or liquid adjuvants, such as stabilisers, for example vegetable oils or epoxidised vegetable oils (e.g. epoxidised coconut oil, rapeseed oil or soybean oil), antifoams, e.g. silicone oil, preservatives, viscosity regulators, binders and/or tackifiers, as well as fertilisers or other active ingredients for obtaining special effects, e.g. acaricides, bactericides, fungicides, nematicides, molluscicides or selective herbicides.

The crop protection compositions according to the invention are prepared in known manner: in the absence of adjuvants, for example by grinding, sieving and/or compressing a solid active ingredient or active ingredient mixture, for example to a specific particle size, and, in the presence of at least one adjuvant, for example by intimately mixing and/or grinding the active ingredient or active ingredient mixture with the adjuvant(s). The invention relates also to those methods of preparing the compositions according to the invention and to the use of the compounds of formulae la and lb in the preparation of such compositions.

The invention relates also to the methods of applying the crop protection products, that is the methods of controlling pests of the type mentioned, such as spraying, atomising, dusting, coating, dressing, scattering or pouring, which are chosen in accordance with the intended objectives and the prevailing circumstances, and to the use of the compositions in the control of pests of the type mentioned. Typical rates of concentration are from 0.1 to 1000 ppm, preferably from 0.1 to 500 ppm, of active ingredient. The rates of application per hectare are generally from 1 to 2000 g of active ingredient per hectare, especially from 10 to 1000 g/ha, preferably from 20 to 600 g/ha.

A preferred method of application in the area of crop protection is application to the foliage of the plants (foliar application), the frequency and rate of application depending on the risk of infestation by the pest in question. The active ingredient may, however, also penetrate the plants through the root system (systemic action) when the locus of the plants is impregnated with a liquid formulation or when the active ingredient is incorporated in solid form, for example in the form of granules, into the locus of the plants, for example into the soil (soil application). In the case of paddy rice crops, such granules may be applied in metered amounts to the flooded rice field.

The crop protection compositions according to the invention are suitable also for the protection of plant propagation material, e.g. seed, such as fruit, tubers or grains, or plant cuttings, against animal pests. The propagation material may be treated with the composition before planting, for example seed may be dressed before being sown. The compounds according to the invention may also be applied to seed grains (coating) either by impregnating the seeds with a liquid formulation or by coating them with a solid formulation. The composition may also be applied to the planting site when the propagation material is being planted, for example may be applied to the seed furrow during sowing. The invention relates also to those methods of treating plant propagation material and to the plant propagation material so treated. The following Formulation and Biological Examples serve to illustrate the invention without, however, limiting the invention to the individual aspects given by way of example. Formulation Examples for use in crop protection (% = percent by weight)

Example F1 : Emulsifiable concentrates a) b) c) active ingredient 25% 40% 50% calcium dodecylbenzenesulfonate 5% 8% 6% castor oil polyethylene glycol ether

(36 mol of ethylene oxide) 5% tributylphenol polyethylene glycol ether

(30 mol of ethylene oxide) 12% 4% cyclohexanone 15% 20% xylene mixture 65% 25% 20%

The finely ground active ingredient and adjuvants are mixed to yield an emulsifiable concentrate from which emulsions of any desired concentration can be obtained by dilution with water.

Example F2: Solutions a) b) c) d) active ingredient 80% 10% 5% 95% ethylene glycol monomethyl ether 20% - - - polyethylene glycol (MW 400) - 70% - -

N-methylpyrrolid-2-one - 20% - - epoxidised coconut oil - - 1% 5% benzine (boiling range: 160-190°) - - 94% -

The finely ground active ingredient and adjuvants are mixed to yield a solution which is suitable for use in the form of microdrops.

Example F3: Granules a) b) c) d) active ingredient 5% 10% 8% 21% kaolin 94% - 79% 54% highly dispersed silicic acid 1 % - 13% 7% attapulgite - 90% _ 18%

The active ingredient is dissolved in dichloromethane, the solution is applied to the carrier mixture by spraying, and the solvent is evaporated off in vacuo. Example F4: Dusts a) b) active ingredient 2% 5% highly dispersed silicic acid 1 % 5% talcum 97% _ kaolin - 90%

The active ingredient and carriers are mixed to yield ready-to-use dusts.

Example F5: Wettable powders a) b) G) active ingredient 25% 50% 75% sodium lignosulfonate 5% 5% - sodium lauryl sulfate 3% - 5% sodium diisobutylnaphthalenesulfonate - 6% 10% octylphenol polyethylene glycol ether - 2% -

(7-8 mol of ethylene oxide) highly dispersed silicic acid 5% 10% 10% kaolin 62% 27% _

The active ingredient and adjuvants are mixed and the mixture is ground in a suitable mill, affording wettable powders which can be diluted with water to give suspensions of any desired concentration.

Example F6: Emulsfiable concentrate active ingredient 10% octylphenol polyethylene glycol ether 3%

(4-5 mol of ethylene oxide) calcium dodecylbenzenesulfonate 3% castor oil polyethylene glycol ether 4%

(36 mol of ethylene oxide) cyclohexanone 30% xylene mixture 50%

The finely ground active ingredient and adjuvants are mixed to yield an emulsifiable concentrate, from which emulsions of any desired concentration can be obtained by dilution with water. Example F7: Dusts a) b) active ingredient 5% 8% talcum 95% - kaolin - 92%

Ready-to-use dusts are obtained by mixing the active ingredient and carrier and grinding the mixture in a suitable mill.

Example F8: Extruder granules active ingredient 10% sodium lignosulfonate 2% carboxymethylcellulose 1 % kaolin 87%

The active ingredient and adjuvants are mixed, the mixture is ground, moistened with water, extruded and granulated, and the granules are dried in a stream of air.

Example F9: Coated granules active ingredient 3% polyethylene gycol (MW 200) 3% kaolin 94%

Uniform application, in a mixer, of the finely ground active ingredient to the kaolin moistened with polyethylene glycol yields non-dusty coated granules.

Example F10: Suspension concentrate active ingredient 40% ethylene glycol 10% nonylphenol polyethylene glycol ether 6%

(15 mol of ethylene oxide) sodium lignosulfonate 10% carboxymethylcellulose 1 % aqueous formaldehyde solution (37%) 0.2% aqueous silicone oil emulsion (75%) 0.8% water 32%

The finely ground active ingredient and the adjuvants are mixed to yield a suspension concentrate, from which suspensions of any desired concentration can be obtained by dilution with water. Biological Examples:

Examples of use in plant protection

Example B1 : Action against Diabrotica balteata larvae

Maize seedlings are sprayed with a formulation comprising 200 ppm of the active ingredient. After the spray-coating has dried, the maize seedlings are populated with 10 Diabrotica balteata larvae in the second stage (L2) and then placed in a plastics container. Evaluation is carried out 6 days later. The percentage reduction in population (% activity) is determined by comparing the number of dead larvae on the treated plants with that on the untreated plants. Cruentaren A is 100% effective against Diabrotica balteata in this test.

Example B2: Action against Tetranychus urticae

Young bean plants are populated with a mixed population (larvae and adults) of Tetranychus urticae and sprayed one day later with a formulation comprising 200 ppm of the active ingredient. The plants are then incubated for 6 days at 25°C and subsequently evaluated.

The percentage reduction in population (% activity) is determined by comparing the number of dead larvae and adults on the treated plants with that on the untreated plants.

The compound cruentaren A is 100% effective against Tetranychus urticae in this test.

Example B3: Action against Frankliniella occidentalis (mixed population, contact action and feeding action)

Discs of bean leaves are placed on agar in petri dishes and sprayed in a spray chamber with a formulation comprising 200 ppm of the active ingredient. The leaves are then populated with a mixed population of Frankliniella occidentalis. Evaluation is carried out 10 days later. The percentage reduction in population (% activity) is determined by comparing the population on the treated leaves with that on the untreated leaves.

The compound cruentaren A is 100% effective against Frankliniella occidentalis in this test.

Example B4: Action against Spodoptera littoralis (larvicide, feeding and contact action)

Discs of cotton leaves are placed on agar in petri dishes and sprayed in a spray chamber with a formulation comprising 200 ppm of the active ingredient. After the spray-coating has dried, the cotton leaf discs are populated with 20-25 Spodoptera littoralis larvae in the first stage (L1 ). 2 and 6 days after treatment, the percentage reduction in population (% activity), repellent action, feeding behaviour and growth regulation are evaluated.

The compound cruentaren A exhibits a very high action (mortality) of 90% against Spodoptera littoralis in this test.

Example B5: Action against Nilaparvata lugens (larvicide. feeding and contact action)

Rice seedlings are sprayed with a formulation comprising 200 ppm of the active ingredient. After the spray-coating has dried, the rice seedlings are populated with 20 Nilaparvata lugens nymphs in the N3 stage. 6 and 12 days after treatment, the percentage reduction in population (% activity), growth regulation and effects on the F1 generation are evaluated.

The compound cruentaren A exhibits an action (mortality) of 100% against Nilaparvata lugens in this test.

Claims

What is claimed is:

1. A compound of the general formula la or lb

or

or a pharmaceutically or agrochemically acceptable salt thereof or any geometric isomer or stereoisomer thereof, or a mixture thereof in enriched form.

2. A compound according to claim 1 having the absolute configuration of cruentaren A or B, obtainable by fermentation of a bacterial strain of the species Byssophaga cruenta.

3. A compound according to claim 2 having the absolute configuration of cruentaren A.

4. A compound according to claim 1 having the absolute configuration of cruentaren A or B in the form of an enriched mixture containing 80-99% by weight, obtainable by fermentation of the production strain Ha r1 of the myxobacterium Byssophaga cruenta.

5. A method of preparing a compound or an enriched mixture according to any one of claims 1 to 4, which comprises culturing the Byssophaga bacterium, and then isolating the metabolite cruentaren A or B either in the form of an enriched mixture or in the form of an isomerically pure individual component.

6. A compound according to either claim 2 or claim 3 designated cruentaren A, characterised by one or more of the following physical parameters:

Mass spectrum: MAT 95 DCI (M + H)⁺ found: 590.37052; calculated: 590.36929.

UV (methanol) λ_max [nm] (logε): 216 (4.26); 265 (3.98); 302 (3.66); 346 (1.94).

IR (KBr) [cm ¹]: 3396, 2959, 2932, 2863, 1646, 1614, 1251 , 1159.

Optical rotation: _D = 3.4 ° (c = 13.3 mg/ml in methanol).

Thin layer chromatography: eluant: ethyl acetate, R_f = 0.53.

Identification by means of analytical HPLC (column: Macherey Nagel 125/2 120 5 μm C-18; eluant: acetonitrile/water 55/45; flow rate: 0.3 ml/minute; detection: diode array):

R_t 3.9 minutes.

¹H-NMR (CDCI₃, 600 MHz, δ [ppm]): 11.48 (broad s, OH); 6.35 (d, 1 H); 6.30 (d, 1 H); 6.08

(broad s, NH); 5.56 (dxdxd, 1 H); 5.49 (1 H); 5.44 (1 H); 5.40 (m, 1 H); 5.29 (dxdxd, 1 H); 3.91

(dxdxd, 1 H); 3.85 (m, 1 H); 3.84 (m, 1 H); 3.80 (s, 3H); 3.75 (dxd, 1 H); 3.64 (dxdxd, 1 H); 3.45

(dxd, 1 H); 2.82 (dxt, 1 H); 2.70 (broad s, 1 H); 2.35 (dxt, 1 H); 2.30 (dxq, 1 H); 2.27 (m, 1 H);

2.24 (m, 2H); 2.22 (m, 1 H); 2.0 (m, 2H); 1.95 (m, 1 H); 1.75 (1 H); 1.56 (broad s, OH); 1.47

(m, 1 H); 1.46 (m, 1 H); 1.33 (m, 1 H); 1.32 (m, 1 H); 1.14 (d, 3H); 1.01 (d, 3H); 0.92 (t, 3H);

0.89 (d, 3H); 0.79 (d, 3H).

¹³C-NMR (CDCI₃, 100 MHz, δ [ppm]): 176.5 (C-23); 171.5 (C-1); 165.8 (C-3); 163.6 (C-5);

143.7 (C-7); 132.2 (C-12); 131.0 (C-20); 126.8 (C-21); 125.8 (C-13); 112.4 (C-6); 105.0

(C-2); 99.7 (C-4); 78.1 (C-15); 74.8 (C-17); 73.1 (C-9); 71.9 (C-25); 55.4 (C-29); 44.9 (C-24);

39.3 (C-16); 38.3 (C-10); 36.9 (C-18); 36.7 (C-8); 36.6 (C-22); 35.8 (C-26); 31.6 (C-11 ); 30.7

(C-19); 29.9 (C-14); 19.3 (C-27); 16.1 (C-32); 14.2 (C-30); 14.1 (C-28); 11.2 (C-33); 8.6

(C-31 ).

7. A compound according to claim 2 designated cruentaren B, characterised by one or more of the following physical parameters:

Mass spectrum: MAT 95 DCI (M + H)⁺ found: 590.37052; calculated: 590.36929. UV (methanol) λ_max [nm] (logε): 210 (4.36); 215 (4.38); 267 (4.14); 301 (3.77). IR (KBr) [cm ¹]: 3416, 2963, 2930, 2869, 1661 , 1628, 1250, 1159. Optical rotation: α_D = 9.1 ° (c = 6.6 mg/ml in methanol). Thin layer chromatography: eluant: ethyl acetate; R_f = 0.40. Identification by means of analytical HPLC (column: Macherey Nagel 125/2 120 5 μm C-18; eluant: acetonitrile/water 55/45; flow rate: 0.3 ml/minute; detection: diode array): R_t 5.0 minutes.

¹H-NMR (CDCI₃, 600 MHz, δ [ppm]): 11.35 (broad s, OH); 6.40 (broad s, 1 H); 6.35 (d, 1 H); 6.26 (d, 1 H); 5.65 (m, 1 H); 5.53 (m, 1 H); 5.50 (m, 1 H); 5.49 (m, 1 H); 4.35 (dxdxd, 1 H); 3.88 (t, 1 H); 3.85 (m, 2H); 3.84 (m, 1 H); 3.80 (s, 3H); 3.47 (dxd, 1 H); 2.93 (dxd, 1 H); 2.80 (dxd, 1 H); 2.35 (m, 1 H); 2.30 (m, 1 H); 2.24 (m, 1 H); 2.22 (m, 1 H); 2.12 (m, 1 H); 2.10 (m, 1 H); 2.02 (m, 1 H); 1.70 (m, 2H); 1.45 (m, 1 H); 1.44 (m, 1 H); 1.32 (m, 1 H); 1.31 (m, 1 H); 1.13 (d, 3H); 1.02 (d, 3H); 0.92 (d, 3H); 0.92 (t, 3H); 0.84 (d, 3H).

¹³C-NMR (CDCI₃, 100 MHz, δ [ppm]): 176.3 (C-23); 170.0 (C-1 ); 165.9 (C-5); 164.6 (C-3); 141.1 (C-7); 131.5 (C-20); 129.8 (C-12); 127.7 (C-13); 126.4 (C-21); 106.4 (C-6); 101.8 (C-2); 99.5 (C-4); 82.3 (C-9); 80.2 (C-17); 77.3 (C-15); 71.9 (C-25); 55.6 (C-29); 44.9 (C-24); 37.3 (C-10); 37.2 (C-16); 36.5 (C-18); 36.2 (C-22); 35.8 (C-26); 33.4 (C-14); 30.7 (C-19); 30.1 (C-11 , C-8); 19.2 (C-27); 15.8 (C-32); 15.1 (C-30); 14.1 (C-28); 11.2 (C-33); 4.4 (C-31 ).

8. A pharmaceutical composition that comprises a compound of formula la and/or lb according to claim 1 together with a pharmaceutical formulation adjuvant.

9. A pharmaceutical composition that comprises the specific isomer cruentaren A and/or B according to claims 2 to 4 together with a pharmaceutical formulation adjuvant.

10. An agrochemical composition that comprises a compound of formula la and/or lb according to claim 1 together with an agrochemical formulation adjuvant.

11. An agrochemical composition that comprises the specific isomer cruentaren A and/or B according to claims 2 to 4 together with an agrochemical formulation adjuvant.

12. A composition according to claims 10 and 1 1 for controlling insects and acarids in crops of useful plants.

13. A method of controlling insects and acarids in crops of useful plants which comprises applying an insecticidally or acaridically effective amount of a compound according to claims 1 to 4 or of a composition comprising such a compound to the insects, acarids or the habitat thereof.

14. A microorganism by means of the fermentation of which a compound of the general formula la or lb can be prepared.

15. A microorganism according to claim 14, which is a microorganism of the order Myxococcales.