CA2097594A1 - Thiangazole, its preparation, compositions and use thereof - Google Patents

Abstract

The invention relates to compounds of general formula (I) especially the compound of formula (Ia) referred to as thiangazole their pharmaceutically acceptable acid addition salts, processes for their preparation and therapeutic compositions and compositions that can be used in crop-protection.

Description

~92/~12~8 pcr/Eps1/o2336 2097~9~ 1 Thiangazole, its preparation. com~ositions and use thereof The present invention relates to compounds of formula I below and, among those, especially the compound of formula Ia called thiangazole, and to the pharmaceutically acceptable acid addition salts of compounds of formulae I and Ia. It relates also to pharmaceudcal compositions (with the excepdon of those against viral diseases) that comprise one of those compounds together with customary formuladon adjuvants.
Compositions that comprise one of the compounds of formula I or la are suitable for controlling Arth~opoda and Acanna and also for controlling parasitdc diseases in humans, animals and plants. Special mendon should be made of their anthelmintic, acaricidal and insecticidal acdvity. In addition to the pharmaceutdcal compositions, the invention there-fore relates also to pesdcides based on compounds of formula I or Ia. It relates also to the preparation of the active ingredients and of the said compositions and to their use for controlling parasites in humans, animals and plants, especially for controlling helminths, especially nematodes, cestodes and trematodes in warm-blooded animals, especially in domestic animals and productive livestock, and to their use for controlling pests of the order Arthropoda, preferably for contTolling insects and representatives of the order Acanna.

The present invention relates especially to compounds of the following general fonnula I
and to their pharmaceudcally acceptable acid addition salts:

~ ~NH

tl) - ~

, :,. .. ~,.~.. . .

WO 92/11258 PCr/EP91/023~ ~
?.~9~ ~9 2-and especially to the compound of forrnula Ia referred to as thiangazole NH
3 3 3 `CH 3 (la) and to its pharrnaceutically acceptable acid addition salts.

That thiangazole is obtainable by culturing the myxobacterium Polyangium spec. DSM
6267 (deposi~ed under E~udapest Treaty with DSM Deutsche Sammmlung von Mikroorganismen und Zellkulturen (Braunschweig, Germany) on December 11, 1990] in a medium containing carbon sources, nitrogen sources and mineral salts, - extracting the harvested cell mass with acetone, - concentrating the (combined) extract, - taking up the concentrate in ether/water, - separating off the aqueous phase and if necessary extracting with ether, - concentrating the (combined) ethereal phase, taking up the concentrate in methanoUheptane, - separating off the methanol phase and if necessary extracting with heptane, ~hen concentrating and extracting with tert-butyl methyl ether, - filtering the extract in tert-butyl methyl ether over Florisil, 92/11258 ~ o 9 7 5 9 4 PCr/EP91/02336 - crystallising thiangazole (or antibiotic andlor fungicidal activity) from the extract and - if appropriate converting the thiangazole (or antibiotic and/or fungicidal activity) into a therapeutically acceptable acid addition salt.

The thiangazole produced by the myxobacterium Polyangium spec. DSM 6267 has the configuration shown in formula Ia. Cis-trans isomers of the thiangazole of the general formula I given above are obtainable by subjecting the thiangazole obtainable by the process indica~ed to heat treatment or to treatment with W light.

All the compounds of the invention can be converted in a manner known Der se into pharmaceutically acceptable acid addition salts.

.
Compounds of formulac I and Ia can also be in the forrn of salts, especially pharmaceuti-cally acceptable, that is to say physiologically tolerable, salts. Pharmaceutically unaccept-able salts can also be used for isolation or purification purposes. Only pharmaceutically acceptable salts are used therapeutically and these are therefore preferred.

Pharmaceutically acceptable acid addition salts are to be understood as being those that are not disadvantagcous physiologically. They are physiologically tolerable addition compounds, especially complexes and salts of a compound of forrnula I with an inorganic or organic base which are formed by the addition of an equivalent amount of a salt-forming base to the basic molecule of formula I or with solvents such as dimcthylforma-mide (D~) or dimethylacetamide (I)MA). There are suitable especially metal salts or ammonium salts, such as alkali metal and alkaline earth metal salts, for example sodium, potassium, magnesium or calcium salts, or ammonium salts with ammonia or with suitable organic amines, especially tertiary monoamines and heterocyclic bases, for example trialkylamines, such as triethylamine or dialkylamines, such as diethylamine or dipropyl-amine, or other organic bascs such as N,N'-dimethylpiperazine.

Furthermore, a preferred embodiment of the invention relates to therapeutic compositions (with the exception of those against viral diseases) that consist of a compound of the : . : , .

' ~

~ogl PCr/EWI/023~ ' invention as active ingredient or comprise such a compound of the invention ~ogether with a customary carrier and/or diluent. Compounds of the invention are to be understood in this contcxt to include pharmaceutically acceptable acid addition salts. Those composi-tions arc suitable for controlling parasidc diseases in humans and animals.

Finally, the invention relates to a composition for plant protection for agriculture, forestry, horticulture and similar areas of use, consisting of thiangazole or comprising thiangazole (if appropriate an agriculturally acceptable acid addition salt) if necessary together with a customary carrier or diluent.

The present invention relates also to a compound, preferably of the thiangazole type, which is characterised by one or more of the parameters given in claim 3.

The invcntion is explained in detail below using expeAmental data:

A. Producdon condiions A.1. Production s;rain: the bacterium Polyangium spec. Strain Pl 3007 order Myxobacteriales, suborder Sorangineae, family Polyandaceae.

A.2. Orivin of production strain: isolated at GBF, Gesellschaft fur Biotechno-logische Forschung mbH (BrunswicklGermany), in October 1986 from soil from Ihe gardens of the Alhambra, Granada, Spain.

A.3. DescriDtion of producdon strain: the vegetative cells are cylindrical with broadly curving ends (Polyangium type), generally about 0.6-0.8 x 4-6 ~lm. In the event of nutrient deficiency, for example when cultured on a smear of living Escherichia coli bacteria on water-agar, the organism forms fruiting bodies. These are plate-like piles of small to medium-sized, spherical or ovoid, gold- to red-brown sporangioles the diameter of which is generally about 30-80 ~m. The sporangiole piles may become very large, but vary greatly in extent, generally from 100 to 600 llm. Pl 3007 grows well on living Escherichia coli bacteria on water-agar, the feedstuff bacteria being broken down. The gliding movement of the bacterial cells causes the colony gradually to spread out in a swarm over the culture plate. In addition, the organism also grows well on yeast-agar (VY/2 agar 0.5 % baker's yeast; 0.1 % CaCI2.2H20; 0.S mgll cyanocobalamine; pH 7.~).
As ;t g~ows, it penetrates deep into the culture substrate and breaks down the yeast cells to .. . . . . . . . . .

. " ` , ' " ~' ' ,. . ~

~ 92/11258 PCr/EP91/02336 - 2097~94 a considerable extent. Catalase and oxidase are positive. In liquid media the strain grows in small clumps of cells both in shaken flasks at 160 rpm (100 ml of medium in a 250 ml Erlenmeycr flask or 500 ml of medium in a 1000 ml Erlenmeyer flask), as well as in bio-reactors (tested up to a sca]e of 300 litres). An example of a suitable culture medium is Poll medium: Probion PS (single-cell protein from Methylomonas clarae; Hoechst, Frank-furt) 0.4 %; starch 0.3 %; MgSO4.7H20 0.1 %; CaCI2.2H20 0.05 %; 1 mUI standard trace-element solution (G. Drews, Messbiologisches Praktikum, 2nd edition, p. 6, Springer Verlag, Berlin 1974) and 1 ml/l standiard vitamin solution (H.G. Schlegel, Allgemeine Mikrobiologie, 6th edition, page 174, Thieme-Verlag, Stuttgart 1985); pH 7Ø The cultures are kept at 30C for 3-4 days. Pl 3007 can be preserved: for example by fieezing vegetative cells from agar plates or liquid cultures in peptone solution at -80C or in liquid nitrogen.

A.4. Performiance of the production striain: An activity can be detected in cell extracts or to a certain extent also in XAD eluates of Pl 3007 that inhibits the growth of certain fungi in an agar diffusion tcst (see below). Chemically the substance consists inter alia of 3 thiazoline nuclei and one oxazole nucleus and has been named thiangazole.

A.S. Availability of the strain: The production strain has been deposited with the Deutsche Sammlung von Mikroorganismen in Brunswick as a patent strain under No. DSM 6267.
A.6. Detection of thiangazole: For qualitadve delecdon of thiangazole cell masses are extracted with acetone or the cultures are stirred with the adsorber resin XAD 1180 (Messrs. Rohm and Haas) and the XAD batches are eluled with methanol and acetoneafter sieving. Aliquots of the concentrated extracts are tested for inhibidon, for example of the fungus Mucor hiemalis, and in a thin-layer chromatography test thiangazole is identified by comparison with the pure substance. The working up, isoladon and chemical characterisation are effected in accordance with Secdon B.

A.7. Production condidons of thiangazole: In shaken flasks thiangazole is formed during growth and reaches its highest activity from the end of the logarithmic phase to the early stationary phase. - -B. Ferrnentation in a bioreactor Bioreactor (bS0) having a capacity of 65 litres from Messrs. Giovanola Frères, Monthey, -';, W0 92/112s8 9 4 pcr/E~l/o23 Switzerland, with two paddle stirrers. Medium: Probion PSC/single-cell protein from Methylomonas clarae; Hoechst, Frankfurt) 0.4 %; starch 0.3 %; MgSO4.7H2O 0.1 %;
CaCI2.2H20 0.05 %; standard trace-element solution 1 mVlitre (see above) and cyano-cobalamine 0.5 mg/litre; pH 7.2. 60 litres of medium are inoculated with 5 litres of culture from a shaken flask having a good culture growth. The aeration rate is set at 200 NVhour and the specd at 200 rpm. The PO2, which is at 100 % saturadon at the start of fermentation, falls continuously to 85 % by the end of fermentation after 9S hours. The pH value rises over the course of the fermentation from 7.0 to 7.2. (Nl denotes normal litre = 1 litre of air under normal conditions.) Isolation of thian azole 309 g of moist Polyangium Pl 3007 cell mass are stirred with 500 ml of acetone. The solid residue is centrifuged off and extracted four times for 30 minutes with approximately 150 ml of acetone each time and then centrifuged off. The combined acetone supernatants are concentrated in vacuo. The residue (6.7 g) is dissolved in ether/water. The aqueous phase is extracted three times with ether and discarded. The combined ethereal phases are concentrated in vacuo (3.54 g) and dissolved in methanoUheptane. After removal of the heptane phase the methanol phase is extracted three dmes with heptane and concentrated in vacuo (1.93 g). The heptane phases are discarded. The crude product from the methanol phase is filtered with S00 ml of tert-butyl methyl ether over 50 ml of Florisil.
The filtrate is concentrated in vacuo and petroleum ether is added. 82 mg of the produc~
are separated in the form of cryst~s. The mother liquor is purified by medium-pressure cluomatography (column (diameter x length) 37 x 420 mm, silica gel 15 Il, 60 A
tHD-SL-15-60, Messrs. Kronwald), eluant petroleum ether/tert-butyl methyl ether/-methanol 50149/1, 28 mUmin, detection W absorpdon at 278 nm). The main peak (tR ~
45 min) is combined and crystallised in tert-butyl methyl ether/petroleum ether (35 mg).
Yield 117 mg.
.
Physical data of thian azole l~etection of thiangazole by analytical HPLC: Column (diameter x length) 4 x 250 mm, packed with HD-S~ 18-5-100 (Messrs. Kronwald), detection W absorption at 290 nm,eluant methanoUwater (80:20). flow 1.5 ml/min, t.~? = 5.8 min; with me~hanoVwater (75~25) tR=10.5 min.-m.p. 140C.-,: . . . . ..

~)9~/llZ58 2097.~9~ PCI/EP91/~2336 UV (methanol)~ /lg ~) = 211 (sh),218 (sh),223 (37884/4.578),228 (sh), 288 (36384/4.561), 300 (sh). (sh = shoulder).-1~ (CHCI3): 3432 (m), 1664 (s),1633 (s), 1577 (m), 1567 (m), 1536 (m), 1465 (w), 1449 (w), 1413 (w), 1370 (w), 1168 (m), 1102 (m), 1015 (m),963 (m) cm~l - (s = s~ong, m =
medium s~ength, w = low intensi~
H-NMR (CDC13,300.133 MHz): ~ = 7.48 (m, 2H),7.35 (M,3H), 7.13 (d, lH, J =
16.2Hz),7.04(d,1H,J=16.2Hz),6.91(mlH),3.85(d,1H,J=11.2Hz),3.81(d,1H,J
= 10.6 Hz), 3.74 (d, lH, J = 11.4 Hz),3.37 (d, lH, J = 11.2 Hz), 3.27 (d, lH, J = 11.4 Hz), 3.20 (d, lH, J = 11.3 Hz),2.93 (d, 3H, J = 5.1 Hz), 2.63 (s, 3H), 1.68 (s, 3H), 1.66 (s, 3H), 1.59 (s, 3H).-3C-NMR (CDCI3,75.473 MHz): ~ = 179.22 s, 178.07 s, 167.90 s, 162.50 s, 162.36 s,153.39 s, 141.99 d, 135.12 s, 129.73 d, 129.18 s, 128.92 d(2C),127.62 d(2C), 122.4& d, 83.69 s, 83.53 s, 79.45 s,43.22 t, 42.50 t, 41.99 t, 26.15 q,25.70 q 25.53 q,24.38 q, 11.75 q. (s, d, t and q denote ~e signal mul~plici~es that would produce an SFORD 13C-NMR
spectrum. 2C denotes doublc signal intensi~y.) -(+)FAB-MS (xenon, ma~lix 3-nilrobenzyl alcohol): m/z = 540 = (M+H)+.-High resolu~on:
[C26H29NSO2S3+Hl calculalcd 540.1562 found 540.1688 (EAB-MS) El-MS (210C,70 eV): m/z (%) = 541 (2.1), 540 (5.1),539 (14) [M ' ],524 (12),493 (16), 379 (4.5), 337 (20), 301 (50),280 (21),260 (44), 213 (23,202 (74), 182 (18), 172 (31), 150 (14), 140 (19), 130 (2''), 115 (21), 103 (10), 85 (22),73 (100).-High resolu~ons:
C26H2gNsO2s3 calc. 539.1483 found 539.1488 (EI-MS) C2SH27NSo2s2 calc. 493.1606 found 493.1604 (EI-MS) Cl4Hl~N4O2s2 calc. 337.0793 found 337.0793 (EI-MS) C,6HI7N2s2 calc. 301.0833 found 301.0833 (EI-MS) C,3HI~N2s2 calc. 260.0442 found 260.0444 (EI-MS) Cl2Hl2NlSl calc. 202.0690 found 202.0691 tEI-MS) ~, ; ~, -x .
, W O 92/11258 PC~r/EP91/0233 ~
~9~94 -8-Commcrcial names HD-Sil-15-60 and HD-Sil-18-5-100 (Kronwald) Florisil (Flolidin Corp.; magnesium silicatc gcl) C. Biolo~ical activitv It has bcen found, suprisingly, that the compounds according to the invention of the general formula I and especially thiangazole according to the invention of formula Ia are -valuable active ingredients against parasites in humans, animals and plants while being well tolcrated by warm-blooded animals, fish and plants. It not only exhibits a broad spectrum of activity against helminths, such as nematodes, cestodes and trematodes, that are parasitcs in the human or animal organism, especially in mammals, its activity bcing dirccted prcferably against nematodes (roundworm), but can also be used successfully against phytopathogcnic insects and arachnids that occur on useful plants and ornamentals in agriculture, especially in cotton, vegetable and fruit crops, in forestry, in the protection of sto~d goods and material stocks, and also in the hygiene sector, especially on domestic animals and producdve livestock. Thiangazole is, moreover, effective against all or individual development stages of normally sensidve and also resistant species of insects and arachnids.

A spccial feature of the compounds of forrnulae I and Ia that should be mentioned is the surprising degree to which they are tolerated by warm-blooded animals, which makes them superior to many other anthelmindcs. The handling of those compounds in practice in the treatrnent of worm-infested animals is facilitated to an extraordinary degree as a result, since even rclatdvely high doses are tolerated by the medicated animals, without any symptoms.

The novel compounds according to the invention of formulae I and Ia are suitable as anthelmindcs for example for controlling parasidc nematodes of the orders (in accordance with K.l. Skrajabin) Rhabdidda Ascaridida Spirurida Trichocephalida or for con~rolling cestodes of the orders (in accordance with Wardle and McLeod)Cyclophyllidae : . . ' ' ~ , . , . ~ :, ' ~92/11258 2097;~9~ pcr/E~l/o2336 Pseudophyllidae or for controlling trematodes of the order .' Digenea in domestic animals and productive livestock, such as cattle, sheep, goats, horses, pigs, red deer, cats, dogs and poultry. Thiangazole can be administered to the animals in a single dose or in repeated doses, the individual doses preferably being from 1 to 50 mg per kg of body weight, according to the species of animal. With protracted adrninistration it is possible in some cases to achieve a better effect or for lower total doses to suffice.

In the case of animal pests from the group of the Insecta and Acarina, the action of the compounds according to the invention of formulae I and Ia may manifest itself in the death of the pests immediately or only at a later date, for exarnple at moulting, or in reduced oviposition andlor a reduced hatching rate. The above-mentioned animal pests include:
of the order Lepidoptera, for example: 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 arnbiguella, 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 spp., Hellula undalis, Hyphantria cunea, Keiferia Iycopersicella, 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 flarnmea, 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, for example: Agriotes spp., Anthonomus spp., Atomaria linearis, Chaetocnema tdbialis, Cosmopolites spp., Curculio spp., Dermestes spp., Diabro~ica spp., Epilachna spp., Eremnus spp., Leptinotarsa decemlineata, Lissorhoptrus spp. Melolontha spp., Orycaephilus spp., Odorhynchus spp., Phlyctinus spp., Popillia spp., Psylliodcs spp., Rhizopertha spp., Scarabeidae, Sitophilus spp., Sitotroga spp.. Tenebrio spp., Tribolium spp. and Trogoderma spp.;
of the order Orthoptera, for example: Blat~a spp., Blattella spp., Gryllotalpa spp., Leucophaea maderae, Locus~a spp.. Periplaneta spp. and Schistocerca spp.;

'' -,' ;

,: ~

WO9~ 8 ~ PCI/EWI/1)23~, ~

of the order Isoptera, for example: Redculitermes spp.;
of the order Psocoptera, for example: Liposcelis spp.;
of the order Anoplura, for example: Haematopinus spp., Linognathus spp. Pediculus spp., Pemphigus spp. and Phylloxera spp.;
of the order Mallophaga, for example: Damalinea spp. and TAchodectes spp.;
Or the order Thysanoptera, for e%ample: Frankliniella spp., Hercinothrips spp., TaeniothAps spp., Thrips palmi, ThAps tabaci and Scirtothrips aurantii;
of the order Heteroptera, for example: 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, for example: Aleurothrixus floccosus, Aleyrodes brassicae, Aonidiella spp., Aphididae, Aphis spp., Aspidiotus spp., Bemisia tabaci, Ceroplaster spp., Chrysomphalus aonidium, Chrysomphalus dictyospermi, Coccus hesperidum, Empoasca spp., Eriosoma laAgerum, Erylhroneura spp., Gascardia spp., Laodelphax spp., Lecanium corni, Lepidosaphes spp., Macrosiphus spp., Myzus spp., Nephotettix spp., Nilaparvata spp., ParatoAa 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 vaporaAorum, TAoza erytreae and Unaspis citri;
of the order Hymenoptera, for example: Acromyrmex, Atta spp., Cephus spp., Diprion spp., Diprionidae, Gilpinia polytoma, Hoplocampa spp., Lasius spp., Monomorium phataonis, Neodiprion spp., Solenopsis spp. and Vespa spp.;
Or the order Diptera, for example: Aedes spp., Antherigona soccata, Bibio hortulanus, Calliphora erythrocephala, Ceratitis spp., Chrysomyia spp., Culex spp., Cuterebra spp., Daeus spp., Drosophila melanogaster, Fannia spp., Gastrophilus spp., Glossina spp., Hypoderma spp., Hyppobosca spp., LiAomyza spp., Lucilia spp., Melanagromyza spp., Musca spp., Oestrus spp., Orseolia spp. Oscinella fAt, Pegomyia hyoscyami, Phorbia spp., Rhagoletis pomonella, Sciara spp., Stomoxys spp., Tabanus spp., Tannia spp. and Tipula spp.;
of the order Siphonaptera, for example: Ceratophyllus spp., Xenopsylla cheopis, of the order Acarina, for example: Acarus siro, AceAa sheldoni, Aculus schlechtendali, Amblyomma spp., Argas spp., Boophilus spp., Brevipalpus spp., Bryobia praeiosa.
Calipitrimerus spp., ChoAoptes spp., Dermanyssus gallinae, Eotetranychus carpini, Eriophyes spp., Hyalomma spp., lxodes spp., Olygonychus pratensis, Ornithodoros spp., Panonychus spp., Phyllocoptruta olei~ora, Polyphago~arsonemus la~us, Psoroples spp., Rhipicephalus spp., Rhizoglyphus spp., Sarcoptes spp., Tarsonemus spp. and Te~ranychus . . .: -, ........ . -. ., . :

. ~ . ~ .......... . . . ...

,. . . . . . . , . - ~ -- .- .. : :- --. .

~ ~ 92/11258 pcr/Ep91/o2336 ~ 2~9759~

spp.; and of the order Thysanura, for example: Lepisma saccharina.

The eompounds aceording to the invention of formulae I and Ia are therefore very suitable for eontrolling pests from the group of the insects and arachnids in cotton, fruit, maize, soybean, citrus and vege~able erops. They control especially plant-eating insects such as Anthonomus grandis, plant-eating insect larvae such as those of Spodoptera littoralis or Heliothis virescens, sucking insects such as Aphis craccivora or Bemisia tabaci and soil insects such as Diabrotica balteata.

The compounds of formulae I and Ia can also be used as dressing agents for protecting seed (fruit, tubers, grains) and plant euttings from noxious inseets and from phytopatho-genie noxious insects whieh occur in the soil.

The invention therefore relates also to eompositions that comprise as active ingredient a compound of formula I or Ia, especially compositions for crop protection, and to their use in the agricultural sector, including agriculture, horticulture and forestry or related fields.

The present invention further embraees the preparation of those compositions which eomprises homogeneously mixing and/or grinding the active ingredient with one or more of the substanees or groups of substanees deseribed herein. The formulation steps can be supplemented by kneading, granulating (in the ease of granules) and, if appropriate, eompressing (in the ease of pellets). Also ineluded is a prophylaede and/or eurative method for eontrolling pes~s of plants and~or helminths in the case of mammals, which eomprises apply ng a eompound aeeording to the invendon of formula I or Ia or the eomposidon aeeording to the invention to the locus of the pest.

Target animals to be proteeted by the use as anthelmintics are all warm-blooded animals that can be infested by helminths, espeeially all mammals ineluding humans, and birds, but especially domestie animals, producdve livestock and pets, such as eows, horses, donkeys, sheep, goats, llamas, eamels, red deer, pigs, dogs, eats, rabbits, hens, turkeys, dueks, geese, pheasants, partridges, etc., as well as all fur-producing farmed animals. It is, of course, possible also ~o treat infested zoo animals successfully.

As is generally known, of the endoparasites occuITing in warm-blooded animals, il is specifically helminths that cause great damage in the animals they infest. The damage .
..

.

. : - . ..

WO 92/11258 PCr/EP91/023~ '~
~9~ 12 caused by helminthiasis can assume very great economic significance where the~ is chronic and especially epidemic occurrence of worrn-related disorders in herds of live-stock. The damage manifests itself in the diseased animals inter alia in reductions in productivity, reduced resistance to other diseases and increased mortality. Especially dangcrous worm-related disorders are brought about by helminths that parasiticise the gas~intestinal tract and other organs and occur relatively frequently, in spite of numerous prophylactic measures, in ruminants, such as cattle, sheep and goats, and also in horses, pigs, poultry, red deer, dogs and cats.

In the present description there is to be understood by the term "helminths" especially parasitic worms that belong to the Platyhelminthes (cestodes, trematodes) and the Nemat-helminthes (nematodes and related species), that is to say tape worms, sucker worms and roundworms of the gastro-intestinal tract and other organs (for example liver, lungs, kidneys, Iymph vessels, blood, etc.).

One of the most pressing tasks is therefore to provide therapeutic compositions that are suitable for controlling helminths in all stages of development and for preventing infesta-tion by those parasites.

Although a number of substances having anthelmintic activity are known that have been proposed for controlling various spccies of helminth, these are not completely satisfactor~ .
either because at a tolerable dose it is not possible to make full use of their spectrum of acivity, or because at therapeutically effective doses they exhibit undesired side effects or properties. In this respect, the resistance to certain classes of substance that is occurring more and more today is also becoming increasingly significant. "Albendazole" which is described, for example, in the literature (British Pat. No. 1464326; Am. J. Vet. Res. 38, 1425-1426 (1977); Am. J. Vet. Res. 37, 1515-1516 (1976); Am. J. Vet. Res. 38, 807-808 (1977); Am. J. Vet. Res. 38, 1247-1248 (1977)) has only a limited spectrum of arithel-mintic activity in ruminants. Its activity against benzimidazole-resistant nematodes and adult livcr flukes is totally inadequate, since in particular the pathogenically important immature migrating forms of the latter are not affected by doses tolerated by the host animal. The provision of compounds having a completely novel basic structure overcomes a large number of resistance problems.

Target crops ~o be protected against phytopathogenic pests within the scope of the present invention by thc crop-protecting use disclosed herein comprise e.g. the following species '. ' , '.' ~' ' ' ' . .. ' : '. : . . ' :.
~' . - ,' ' '~. .' '' ~ ' ' ' '.
'.
~- ' .' . ' :
r, ' : ' ` '' . ' ; : .~
,, ' .

~ 92/11258 2 0 9 7 ~ 9 4 pcr/Epsl/o2336 of plants: cereals (wheat, barley, rye, oats, rice, maize, sorghum and related crops), beet (sugar beet and fodder beet), pomes, drupes and soft fruit (apples, pears, plums, peaches, almonds, chcrries, strawberries, raspberries and blackberries), leguminous plants (beans, lentils, peas, soybeans), oil plants (rape, mustard, poppy, olives, sunflowers, coconut, castor oil plants, cocoa beans, groundnuts), cucumber plants (cucumber, marrows,melons), fibre plants (cotton, flax, hemp, jute), citrus fruit (oranges, lemons, grapefruit, mandarins), vegetables (spinach, lettuce, asparagus, cabbages, carnots, onions, tomatoes, potatoes, paprika), lauraceac (avocados, cinnamon, camphor), or plants such as tobacco, nuts, coffee, aubcrgincs, sugar cane, tea, pepper, vines, hops, bananas and natural rubber plants, as well as omamentals.

The compounds of fommulae I and Ia are nommally applied in the form of compositions and can bc applicd to the crop area or plant to be treated, simultaneously or in succession, with furthcr compounds. Thcsc funher compounds can be fertilisers or micronutrient donors or othcr preparations that influence plant growth. They can also be selective herbicides as well as insecticides, fungicides, bactericides, nematicides, molluscicides or mixtures of scvcral of these prcparations, if desired together with further carricrs, surfactants or other application-promoting adjuvants customarily employed in fommulation technology.

The good pesticidal activity of the compounds of formulae I and la according to the invention corresponds to a mortality of at least S0-60 % of the mentioned pests.
The activity of thc compounds of the invcntion and of the composidons comprising them against animal pests can be substantially broadened and adapted to prevailing circum-stances by the addition of other insecticides and/or acaricides. Examples of suitable additives include rep~sentatives of the following classes of compounds: organo-phosphorus compounds, nitrophenols and derivatives thereof, fommamidines, ureas,carbamates, pyrethroids, chlorinated hydrocarbons, and Bacillus thuringicnsis preparations.

The compounds of formulae I and la are used in umnodified form or, preferably, together with thc adjuvants convendonally employed in formulation technology, and can therefore be formulated in known manner e.g. into emulsifiable concentrates, directly sprayable or dilu~able solutions, dilute emulsions, wet~able powders, soluble powders, dusts, granules.
and also encapsuladons in polymer substances. As with the composidons. the methods of application, such as spraying, atomising, dusting, scattering or pouring, are chosen in i . , -. - - . .

,:
- :~
., '. ' -.

WO 92/11258 PCr/EP91/023~ , ~
~,~9~ ~9 - 14 -accordance with the intended objectives and the prevailing circumstances.

A prefcrred method of applying a compound of formula I or Ia, or an agrochemicalcomposidon which comprises at least one of said compounds, is foliar application. The number of applications and the rate of application depend on the risk of infestation by the corrcsponding pathogen. Howcver, the compounds of formulae I and Ia can also penetrate the plant through the roots via the soil (systemic acdon) if the locus of the plant is impreg-nated with a liquid formulation, or if the compounds are applied in solid form to the soil, -e.g. in granular form (soil application). In paddy rice crops, such granules may be applied in metered amounts to the flooded rice field. The compounds of formulae I and Ia may, however, also be applied to seeds (coadng) either by impregnadng the seeds with a liquid formulaion comprising the compound, or by coatdng them with a solid formulation.
The formulations, i.e. the composidons, comprising as active ingredient a compound of the invention, or combinadons of those compounds, and, where appropriate, a solid or liquid adjuvant, are prepared in known manner, e.g. by homogeneously mixing and/or grinding the acdve ingredients with extenders, e.g. solvents, solid carriers and, where appropriate, suriface-acdve compounds (surfactants).

Suilable solvents are: aromadc hydrocarbons, preferably the C8 to Cl2 fractions of aLlcyl-benzenes, e.g. xylene mixtures or alkylated naphthalenes, aliphadc or cycloaliphatic hydrocarbons such as cyclohexane, paraffins or tetrahydronaphthalene, alcohols such as ethanol, propanol or butanol, and glycols and their ethers and esters, such as propylene glycol, dipropylene glycol ethcr, ethylene glycol, ethylene glycol monomethyl or mono-ethyl ethcr, ketones such as cyclohexanone, isophoronc or diacetone alcohol, strongly polar solvents such as N-methyl-2-pyrrolidone, dimethyl sulfoxide or dimethylformamide, or water, vegetable oils such as rapeseed oil, castor oil, coconu~ oil or soybean oil; and, whero appropriate, silicone oils.

The solid carriers used, e.g. for dusts and dispersible powders, are norrnally natural mineral fillers such as calcite, talcum, kaolin, montmorillonite or attapulgite. In order to improve the physical properdes it is also possible to add highly dispersed silicic acids or highly dispersed absorbent polymers. Suitable granulaud adsorpdve carriers are porous types, for example pumice, broken brick, sepiolite or bentonite; and suitable nonsorbent carriers are calcite or sand. In addidon, a great number of granulated materials of inorganic or organic nature can be used, e.g. especially dolomite or pulverised plam . .
-.... ~ , .
~ . - ' ' ., . : . .

' ' ' ' ' :
. ,f..

: , : - .

~9Z/112S8 PCI/EWI/02336 2097~9~ 1~

residues.

Suitable surface-acdve compounds are the combinations of of the compounds of formula I
or Ia with other insecticides or acaricides together with non-ionic, cationic and/or anionic surfactants having good emulsifying, dispersing and wetting properties. The term"surfactants" will also be understood as comprising mixtures of surfactants.

Both so-called water-soluble soaps and water-soluble synthetic surface-active compounds are suitablc anionic surfactants.

Suitable soaps are the alkali metal salts, alkaline earth metal salts or unsubstituted or substituted ammonium salts of higher fatty acids (C1o-C22), e.g. the sodium or polassium salts of oleic or stearic acid, or of natural fatty acid mixtures which can be obtained e.g.
from coconut oil or tall oil. Mention may also be made of fatty acid methyltaurin salts as surfactants.

More frequently, however, so-called synthetic surfactants are used, especially fatty sulfonates, fatty sulfates, sulfonated benzimidazole derivatives or aL~cylarylsulfonates.

The fatty sulfonates or sulfates are usually in the form of alkali metal salts, alkaline earth metal salts or unsubsdtuted or substituted ammonium salts and generally contain a C8-C22-alkyl radical, which also includes the allcyl moiety of acyl radicals, e.g. the sodium or ealeium salt of lignosulfonic acid, of dodecyl sulfate or of a mixture of fatty alcohol sulfates obtained from natural fatty acids. These compounds also comprise the salts of sulfated and sulfonated fatty alcohoVethylene oxide adducts. The sulfonated benzimid-azole derivati~es preferably contain 2 sulfonic acid gsoups and one fatty acid radical containing approximately 8 to 22 carbon atoms. Examples of alkylarylsulfonates are the sodium, calcium or triethanolamine salts of dodecylbenzenesulfonic acid, dibutylnaphtha-lenesulfonic acid, or of a condensate of naphthalenesulfonic acid and foTrnaldehyde. Also suitable are corresponding phosphates, e.g. salts of the phosphoric acid ester of an adduct of p-nonylphenol with 4 to 14 moles of ethylene oxide, or phospholipids.

Non-ionic surfactants are especially polyglycol ether derivadves of aliphatic or cyclo-aliphatic alcohols, saturated or unsaturated fatty acids and alkylphenols, said derivatives containing 3 to 30 glycol ether groups and 8 to 20 carbon atoms in the (aliphatic) hydro-carbon moiety and 6 to 18 carbon atoms in the alkyl moiety of the alkylphenols. Further - . ; ;-WO92/11258 PCr/EP91/023~
?.~9~ ~9 4 - 16-suitable non-ionic surfactants are the watersoluble adducts of polyethylene oxide with polypropylenc glycol, ethylenediaminopolypropylene glycol and alkylpolypropyleneglycol containing I to 10 carbon atoms in the alkyl chain, which adducts contain 20 to 250 cthylcnc glycol ether groups and 10 to 100 propylene glycol ether groups. These compounds usually contain 1 to 5 ethylene glycol units per propylene glycol unit.

Representadve examples of non-ionic surfactants are nonylphenolpolycthoxyethanols, castor oil polyglycol ethers, polypropylene/polyethylene oxide adducts, tributylphenoxy-polyethoxycthanol, polyethylene glycol and octylphenoxypolyethoxyethanol. Fatty acid esters of polyoxyethylene sorbitan, e.g. polyoxyethylene sorbitan trioleate, are also suitablc non-ionic surfactants.

Cationic surfactants are especially quatemary ammonium salts which contain, as N-subsdtuent, at least one CB-C22aL~yl radical and, as further subsdtuents, unsubstituted or halogcnated lower alkyl, benzyl or hydroxy-lower alkyl radicals. The salts are preferably in the form of halides, methyl sulfates or ethyl sulfates, e.g. stearyltrimethylammonium chloride or benzyldi(2-chloroethyl)ethylammonium bromide.

The surfactants customarily employed in formulation technology are described, for examplc, in the following publications:

"McCutcheon's Detergents and Emulsifiers Annual", MC Publishing Corp., Glen Rock, NJ, USA, 1988", H. Stache, "Tensid-Taschenbuch", 2nd ediion, C. Hanser Verlag, Munich, Vienna, 1981, M. and J. Ash, "Encyclopedia of Surfactants", Vol. I-m, Chemical Publishing Co., New York, 1980-1981.

The pesticidal compositions for crop protection usually comprise 0.1 to 99 %, especially 0.1 to 95 %, of a compound of formula I or forrnula Ia or combinations of that compound with other insecticides or acaricides, 1 to 99.9 % of a solid or liquid adjuvant, and 0 to 25 %, especially 0.1 to 25 %, of a surfactant. Whereas commercial products will preferably be formulated as concentrates, the end user will normally employ dilute formulations comprising considerably lower active ingredient concentraions. Typical appiicaion concentrations are from O. 1 to 1000 ppm, preferably from 0.1 to ~OO ppm. For , . ~ . . .
.
.- - : ' ' :.

. . : .~ .. : . ' . .' ' :
. .

~ 92/11258 PCr/EP91/02336 - 2097S9~

crop protection, the rates of application per hectare are generally from l to 2000 g of active ingredient per hectare of cultivated area, preferably from 10 to lO00 g/ha, cspecially from 20 to 600 g/ha.

Preferred forms of administration to warm-blooded animals for controlling helminths include solutions, emulsions, suspensions (drenches), feed additives, powders and tablets, including effervescent tablets, boli, capsules and micro-encapsulations, it being necessary to take into account the physiological acceptability of the formulation adjuvants.

Suitable binders for tablets and boli are chemically modified natural polymer substances that are soluble in water or alcohol, such as starch, ceDulose or protein derivatives (e.g.
methylcdlulose, carboxymethylccllulose, ethylhydroxyethylcellulose, proteins, such as zein, gelatin and the like) and synthetic polymers, such as, for example, polyvinyl alcohol, polyvinylpyrrolidone etc.. The tablets also comprise fillers (e.g. starch, microcrystalline ceDulose, sugar, lactose etc.), glidants and disintegrators.

If the anthelmindc composidons are in the form of feed concentratcs, then the carriers used are, for example, performance fecd, fecd grain or protcin concentrates. In addidon to the acdve ingredients, such fccd concentrates or compositions may contain additives, vitamins, andbiotdcs, chemothcrapeudc agents, or other pesdcides, cspecially bacterio-stadcs, fundstadcs or coccidiostatics, or also hormonc preparatdons, substanccs having an anabolic acdvity, or substanccs that promote growth, influence the mcat quality of animals for slaughter or are useful to the organism in some other way. If thc compositions or the active ingredients of for nula I they comprise are added directly to thc fced or to the herd Wnks, then the prepued feed or the prepared Wnk preferably compriscs thc acdve ingredients in a concentradon of from approximatcly 0.0005 to 0.02 pcrccnt by wcight (5-200 ppm).

The compositions according to the invendon can be administered to the animals to be treated perorally, parenterally or subcutaneously, the compositions bdng in the form of soludons, emulsions, suspensions (drenches), powders, tablets, boli and capsulcs.

The anthelmintic compositions according to the invention gcncrally comprise from 0.1 to 99 % by weight, preferably from 0.1 to 95 % by weight, of a compound of forrnuia 1, la or mixtures thereof, from 99.9 to I % by weight, preferably from 99.8 to 5 % by weight, of a solid or liquid adjuvant, including from 0 to 25 % by weight, preferably from 0.1 to 25 %

: .

WO 92/11258 " 9 4 - 18 - P~/EP91/02 by weight, of a surfactant.

Whercas commercial products will prcferably be formulated as concentrates, the end user will normally cmploy dilute formulations.

Thc composidons may also comprise further auxiliaries such as stabilisers, antifoams, viscosity regulators, binders, tackifiers as well as other active ingredients for obtaining special effects.

The present invention relates also to such anthelmintic compositions employed by the end user.

In each of the methods for pest control according to the invention and in each of the pcsticides according to the invention the compound of formula I may be used in any of its structural configurations, in mixtures thereof or in the form of its salts.

The invention includes also a method for the prophylactic protection of warm-blooded animals, especially productive livestock, domestic animals and pets, against parasitic helminths, which comprises administering the compound of formula I or active ingredient forrnulations prepared therefrom to the animals in the form of an additive 10 the feed or to the drinl; or, alternatively, in solid or liquid forrn, orally, by means of injection or parenterally. The invention includes also the compounds according to the invention of forrnula I for usc in one of the mentioned methods.

The Exarnples that follow serve mcrely to illus~ate the invention without limiting the invendon.
Preferred formuladons have especially the following composition: (throughout, percent-ages ~re by weight) Ernulsifiable concentrates:
thiangazole l to 90 %, preferably S to 20 %
surfactant: 1 to 30 %, preferably lO to 20 %
liquid carrier: S to 94 %, preferably 70 to 85 %

. : - : . - . .. .: . .. . . . .

.:
. , . : , . .

~

~ 9~/11258 2 (~ 9 7 ~ 9 4 PCI/EP91/l)Z336 ¦ -Dusts: j thiangazole 0.1 to 10 %, preferably 0.1 to 1 %
solid carrier: 99.9 to 90 %, preferably 99.9 to 99 %

Suspension conccntrates:
thiangazole S to 75 %, prcferably 10 to 50 %
water 94 to 24 %, preferably 88 to 30 %
surfactant: 1 to 40 %, preferably 2 to 30 %

Wettable powders:
thiangazole 0.5 to 90 %, preferably I to 80 %
surfactant: 0.5 to 20 %, preferably 1 to 15 %
solid carrier 5 to 95 %, preferably 15 to 90 %

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

The compositdons may also comprise further addidves such as stabilisers, for example vegetable oils or epoxidised vegetable oils (epoxidised coconut oil, rapeseed oil or soybean oil), andfoams, for exarnple silicone oil, preservatives, viscosity regulators, binders, tacldfiers as well as fertilisers or other active ingredients for obtaining special effects.

W092/11258 9~594 PCr/EP9l/023~

The following Examples serve to illustrate the invention. They do not limit the invention.

D. Preliminarv tests for biolo,eical activitv The antibiotic activity is detcrmined by means of an agar diffusion test with reference to -the inhibiting areola diameter, or from the culture density in a series dilution test. Thian-gazole inhibits the growth of some fungi, for example Mucor hiemalis, Botrytis cinerea, Gibberella fujikuroi, Rhizopus arrhizus and Pythium debaryanum. The MIC for Ustilago maydis is 3.2 llg/ml. In the case of sub-mitochondrial particles of deer heart, thiangazole inhibits NADH oxidation. Lnvestigations by differential spectroscopy show complex I
(NADH2: ubiquinone-oxidoreductase) of the eukaryotic respiratory chain to be the site of action of thiangazole.

E. Formulation Examples E. l . Emulsifiable concentrates a) b) c) a compound of formula Ia 25 % 40 % S0 %
calcium dodecylbenzenesulfonate 5 % 8 % 6 %
castor oil polyethylene glycol ether (36 moles of ethylene oxide) S %
tributylphenol polyethylene glycol ether (30 moles of ethylene oxide) - 12 % 4 %
cyclohexanone - 15 % 20 %
xylene mixture 65 % 25 % 20 %

Emulsions of any desired concentration can be produced from such concentrates bydilution with water.
E.2. Emulsifiable concentrates a) b) c) a compound of formula Ia 10 % 8 % 60 %
octylphenol polyethylene glycol ether (4-5 moles of ethylene oxide) 3 % 3 % 2 ~0 calcium dodecylbenzenesulfonate 3 % 4 % 4 %
castor oil polyethylene glycol ether (35 moles of ethylene oxide) 4 % 5 % 4 %
cyclohexanone 30 % 40 % lS ~c xylenc mixture 50 % 40 % 15 %

. : -. , : . .
.. .
. - ~ . ~, . . .

"., '` '`' - ' : ~ - . . , . -- .

9~/11258 ~! 0 9 7 ~ 9 1 pcr~Ewl/o2336 Emulsions of any desired concentration can be produced from such concentrates bydilution with water.

E.3. SusPension concentrate a compound of formula Ia 40 %
ethylene glycol 10 %
nonylphenol polyethylene glycol ether (15 moles of ethylene oxide) 6 %
sodium lignosulfonate 10 %
carboxymethylcellulose 1 %
37 % aqueous formaldehyde solution 0.2 %
silicone oil in the form of a 75 %
aqueous emulsion 0.8 %
water 32 %

The finely ground active ingredient is intimately mixed with the adjuvants, affording a suspension concentrate from which suspensions of any desired concentration can be obtained by dilution with water.

E.4. Water-dispersible powder mixtures a) b) c) a compound of fonnula la 25 % 50 % 75 %
sodium lignosulfonate 5 % 5 %
oleieacid 3% - 5 %
sodium diisobutylnaphthalene-sulfonate - 6 % 10 %
octylphenol polyethylene glycol ether (7-8 moles of ethylene oxide) - 2 %
highly dispersed silicic acid 5 % 10 % 10 %
kaolin 62 % 27 % - -The active ingredient is thoroughly mixed with the adjuvanls and the mixture is thoroughly ground in a suitable mill, affording wettable powders which can be diluled with wa~er to give suspensions of any desired concentration.

.: .; , ., . . . ~, ..

2~9rl ~ pcr~Eps1/o23~

E.5. Dusts a) b) a compound of formula Ia 2 % 5 %
highly dispersed silicic acid1% 5 %
tatcum 97 %
kaolin - 90 %

Ready-for-use dusts are obtained by intimately mixing the carriers with the active ingredient and grinding the mixture.

E.6. Granules a) b) a compound of formula Ia 5 % lO %
kaolin 94 %
highly dispersed silicic acid1 %
attapuldtc - 90 %

The active ingredient is dissolved in methylene chloride, the solution is sprayed onto the carrier, and the solvent is subsequently evaporated off in vacuo. Such granules can be mixed with anirnal feed.

E.7. Granules a compound of formula Ia lO %
sodium lignosulfonate 2 %
carboxymethylcellulose I %
kaolin 87 %

The tctive ingredient is mixed and ground with the adjuvants, and the mixture issubsequently moistened with water. The mixture is extruded and then dried in a stream of air.

E.8. Granules a compound of formula la 3 %
polyethylene glycol (mol. wt. 200) 3 %
kaolin 94 %

The finely ground active ingredient is uniformly applied, in a mixer, to the kaolin mois~ened with polyethylene glycol. Non-dusty coated granules are obtained in this .... . . . .
, . .. , , - . .. ~ . - . - . , .. . . , ~ .. .. . . .
.. .. . . . ~ .
. .. . . -.

, - - :: , : , . -: . , .

~? 92/11258 2 0 9 7S9 ~ PCr/EP91/02336 manner.

E.9. Tablets or boli I compound of formula Ia 33.00 %
methylccllulose 0.80 %
highly dispersed silicic acid 0.80 %
cornstarch 8.40 %
t II crystalline lactose 22.50 %
cornstarch 17.00 %
microcrystalline cellulose 16.50 %
magnesium stearate 1.00 %

The methylcellulose is stilred into water and allowed to swell; the silicic acid is stirred in and the mixture is made into a homogeneous suspension. The active ingredient and cornstarch are mixed and the aqueous suspension is incorporated into this mixture which is kneaded to a paste. The mass so obtained is granulated through a 12M sieve and dried.

II All four adjuvants are thoroughly mixed.

III The premixtures obtained in accordance with I and II are mixed and compressed to tablets or boli.

E.10. Iniectable DreDarations a. Oilv vehicle tslow release~
compound of formula la 0.1-1.0 g groundnut oil ad 100 ml compound of formula Ia 0.1-1.0 g sesame oil ad 100 ml Preparation: The active ingredient is dissolved in some of the oil with sti~ing and if necessary with gentle heating. After cooling, the solution is made up to the desired . . .

WO 92/11258 pcr/Ep91/o23~ ,~
~9rl ~9 24 volume and sterile-filtered through a suitable 0.22 llm membrane filter.

Q. Water-miscible solvent (medium rate of release) compound of forrnula Ia 0.1-1.0 g 4-hydroxymethyl- 1 ,3-dioxolane (glycerol formal) 40 g 1,2-propanediol ad 100 ml compound of formula la 0.1-1.0 g glycerol dimethyLtcetal 40 g 1,2-propanediol ad 100 ml Preparation: Thc active ingredient is dissolved in some of the solven~ with stirring, made up to the desircd volume and sterile-Sltered through a suitable 0.22 llm membrane filter.

y. Aqueous soluble preparation (rapid release) compound of formula la 0.1-1.0 g polyethoxylated castor oil (40 ethylene oxide units) 10 g 1,2-propanediol 20 g benzyl alcohol I g Aqua ad inject. ad 100 ml compound of formula Ia 0.1-1.0 g polycthoxylated sorbitan monooleate (20 ethylene oxide units) 8 g 4-hydroxymethyl- 1,3-dioxolane (glycerol forrnal) 20 g benzyl alcohol I g Aqua ad inject. ad 100 ml Preparation: The active ingredient is dissolved in the solvents and the surfactant, made up to the desired volume with water and then sterile-filtered through a suitable membrane filter having a pore diameter of 0.22 llm.

The aqueous systems can be used preferably also for oral and/or intr~rumin31 adminis-tration.

' , ', - ;, .
' ~ 92~11258 pcr/Ep9l~o2336 2097~9~

F. Biolo~ical Examples The anthelmintic activity is demonstrated by means of the following tests:

F. 1. Trial with sheeP infested with nematodes such as Haemonchus contortus and Tricho-stron~lus colubriformis Thiangazole is administered in the form of a suspension using a stomach probe or by intra-ruminal injection to sheep that have previously been artificially infested with nematodes, such as Haemonchus contortus and Trichostrongylus colubriformis. l to 3 animals are used for cach dosc per trial. Each sheep is treated only once with a single dose.

A first evaluati~n is made by comparing the number of worm eggs excreted in the faeces of the sheep before and after treatment.

Seven to ten days after treatment the sheep are sacrificed and dissected. The evaluation is carried out by counting the worms remaining in the intestine after the treatment. Sheep simultaneously and similarly infested but untreated are used as a control or comparison.

A sharp reduction h nematode infestation is achievcd with thiangazole in this trial. For example the use of 20 mg of active ingredient per kg of body weight effects virtually complete reduction of nematode infestation.

F.2. Trial with sheeD infested wdh cestodes such as Moniczia benedeni The active ingredient is administered in the form of a suspcnsion using a stomach probe or by intraruminal injection to sheep that have prcviously been artificially infested with cestcdes, such as Moniezia benedeni. 1 to 3 animals are used for each dose per trial. Each sheep is treated only once with a single dose. Seven to ten days after treatment the sheep are sacrificed and dissected. The evaluation is carried out by counting the worms remaining in the intestine after the treatrnent. Sheep simultaneously and similarly infested but untreated are used as a control or comparison. In this test thiangazole effects an approximately 90 % reduction in cestode infestation at a dose of less than 20 mg/lcg bodv weight.

: . :, ,, . -.
- ' ' ' ; ' -: ", "................. .. ' . :' ~", ' ' ' : ., ~ . . .

o 3 pcr/Ep9l /023 F.3. Action against Derrnanvssus allinae 2 to 3 ml of a solution comprising 100 ppm of test compound, and approxunately 200 mites at vaAous stages of development, are placed in a glass container that is open at the top. The container is then closed with a cotton wool plug, shaken for 10 minutes until the mitcs are completely wetted, and then inverted for a short time so that the remaining test solution can be absorbed by the cotton wool. After 3 days, the mortality of the mites is determined as a percentage by counting the number of dead individuals. Thiangazole exhibits good activity against Dermanyssus gallinae in this-test.

F.4. Action a~ainst Lucilia sericata Freshly deposited eggs of the blow fly Lucilia seAcata are in~oduced in small portions (30-50 eggs) into test tubes in which 4 ml of nutrient medium have previously been mixed with I ml of test solution in the intermediate dilution required for the final concentration.
After inoculation of the culture medium the test tubes are closed with a cotton wool plug and incubated in an incubator at 30C for four days. By this time, larvae approximately I cm long (stage 3) have developed in the medium that, for comparison, has not been treated. If a compound is active, the larvae are at this time either dead or moribund and clearly retarded. The test is carried out simultaneously at concentrasions of from 10 -0.01 ppm. The activity is measured at the lowest fully active concentration (LC 100). The test includes both compounds that are effective by contact and those that are effective as an ingested poison. Repellence is also taken into account, since the larvae migrate from the medium and starve. Thiangazole exhibits full activity even at low concentrations in this test.

F.S. Action a~ainst Lucilia cuPrina Freshly deposited eggs of the blow fly species Lucilia cuprina are introduced in small portions (30-S0 eggs) into test tubes in which 4 ml of nutrient medium have previously been mixed with I ml of test solution comprising 16 ppm of the test compound. After inoculation of the culture medium the test tubes are closed with a cotton wool plug and incubated in an incubator at 30C for four days. By this time, larvae approximately I cm long (stage 3) have developed in the untreated medium. If a compound is active the larvae are at this time either dead or clearly retarded. Evaluation is effected after 96 hours.
Thiangazole exhibits very good activity against Lucilia cuprina.

: ' :
.

~ 92/11258 2 ~ 9 7~ 9 ~ PC~r/EP91/02336 t F.6. Trial with sheep infested with Fasciola hepatica The activc ingredient is ad ninis~ered in the form of a suspension using a stomach probe or by intraruminal injection to sheep that have previously been arti~lcially infested with Fasciola hepatica 3 animals are used for each dose per trial. Each animal is treated only once with a single dose.

A first evaluation is made by comparing the number of worm eggs excreted in the faeces of the sheep before and after treatment. ~ -Three to four weeks after trea~ment the sheep are sacrificed and dissected. The evaluation is carried out by counting the liver flukes remaining in the gall-bladder ducts after treat-ment. Sheep simultaneously and similarly infested but untreated are used as a control or comparison. The difference in the number of liver flukes counted in the two groups gives .
the degree of cffectivcness of thc test compound.

Thiangazolc cxhibits good activity against Fasciola hepadca at doses of less than 50 mg of acdve ingredient/kg of body weight in this test.
.

F.7. OvicidaUlarvicidal action a~ainst Heliothis virescens Egg deposits of Heliothis viresccns on cotton are sprayed with an aqueous emulsion comptising 400 ppm of test compound. 8 days later, the percentage of eggs which have hatched and the survival rate of the caterpillars are evaluated in comparison with untreated controls (% reducdon in the population). Thiangazole exhibits good activity against Heliothis virescens in this test.

F.8. Action a,eainst ADhis cracci_ora Pea seedlings are infested with Aphis craccivora and then sprayed with a spray mixture comprising 400 ppm of the test compound, and incubated at 20C. Evaluation is made 3 and 6 days later. The percentage reduction in the population (% activity) is detennined by comparing the number of dead aphids on the treated plants with that on untreated plan~s. Thiangazole exhibits good activity against Aphis craccivora in this test.

4 PCl /EP91/023 F.9 Action a~ainst Tetranychus urticae Young bean plants are populaled with a mixed population of Tetranychus urticae and sprayed one day later with an aqueous emulsion comprising 400 ppm of the test compound. The plants are then incubated for 6 days at 25C and then evaluated. The percentage reduction in the population (% activity) is determined by comparing the number of dead eggs, larvae and adults on the treated plants with that on untreated plants.
Thiangazole exhibits good activity against Tetranychus urticae in this test.

F.10. Action a~ainst Heliothis virescens caterpillars Young soybean plants are sprayed with an aqueous emulsion comprising 400 ppm of the test copound. After the spray coating has dried, the soybean plants are populated with 10 Heliothis virescens caterpillars in the f~st stage and placed in a plastics container.
Evaluation is made 6 days later. The percentage reduction in the population or the percentage reduction in feeding damage (% activity) is determined by comparing the number of dead caterpillars and the feeding damage on the treated plants with that on untreated plants. Thiangazole exhibits very good activity against Heliothis virescens caterpillars.

F.l I . Action a~ainst Plutella xvlostella caterDillars Young cabbage plants are sprayed with an aqueous emulsion comprising 400 ppm of the test compound. After the spray coating has dried, thc cabbage plants are populated with 10 Plutella xylostella caterpillars in thc third stage and placed in a plastics container.
Evaluation is made 3 days later. The percentage reduction in the population or the percentage r;educdon in feeding damage (% activity) is determined by comparing the number of dead caterpil~ars and the feeding damage on the treated plants with that on untreated plants. Thiangazole exhibits good activity against Plutella xylostella in this test.
The activity is ovcr 80 %.

' :"~ "- ' ' ' ~ . ' .

Claims (12)

WO 92/11258 PCT/EP91/0233?

What is claimed is:

1. A compound of the following general formula I or a pharmaceutically acceptable acid addition salt thereof:

(I)

2. A compound according to claim 1 having the absolute configuration given underformula Ia (Ia) or a pharmaceutically acceptable acid addition salt thereof.

3. A substance obtainable from the culture broth of the myxobacterium Polyangium spec.
DSM 6267, having one or more of the following parameters, or a pharmaceutically acceptable acid addition salt thereof:
m.p. 140°C
UV (methanol):.lambda.max(.epsilon./lg .epsilon.) = 211 (sh),218(sh), 223(37884/4.578), 228 (sh), 288 (36384/4.561), 300 (sh). (sh = shoulder).-IR (CHCl3): 3432 (m), 1664 (s), 1633 (s), 1577 (m), 1567 (m), 1536 (m), 1465 (w), 1449 (w), 1413 (w), 1370 (w), 1168 (m), 1102 (m), 1015 (m), 963 (m) cm-1 - (s = strong, m =
medium-strength, w = low intensity).-?H-NMR (CDCl3, 300.133 MHz): .delta. = 7.48 (m, 2H),7.35 (M, 3H), 7.13 (d, 1H, ? =
16.2 Hz), 7.04 (d, 1H,? = 16.2 Hz), 6.91 (m 1H), 3.85 (d, 1H, ? = 11.2 Hz), 3.81 (d, 1H.?
= 10.6 Hz), 3.74 (d, 1H, ? = 11.4 Hz), 3.37 (d, 1H, ? = 11.2 Hz), 3.27 (d, 1H, ? = 11.4 Hz), ?O 92/11258 -30- PCT/EP91/02336 3,20(d,1H,?=11.3 Hz),2.93(d,3H,?=5.1Hz),2.63(s,3H),1.68(s,3H),1.66(s,3H), 1.59(s,3H).-13C-NMR(CDCl3, 75.473 MHz):.delta.=179.22s, 178.07s, 167.90s, 162.50s, 162.36s, 153.39s, 141.99d, 135.12s, 129.73d, 129.18s, 128.92d(2C), 127.62d(2C), 122.48d, 83.69s, 83.53s, 79.45s, 43.22t ,42.50t, 41.99t, 26.15q, 25.70q 25.53q, 24.38q, 11.75 q.(s,d,t and q denote signal multiplicities that would produce an SFORD 13C-NMR
spectrum.2C denotes double signal intensity.) (+)FAB_MS(xenon,matrix 3-nirobenzylalcohol):m/z=540=(M+H)+.-High resolution;
[C26H29N5O2S3+H]calculated 540.1562 found 540.1688(FAB-MS) EI-MS(210°C,70eV):m/z(%)=541(2.1),540(5.1),539(14)[M+],524(12),493(16), 379(4.5),337(20),301(50),280(21),260(44),213(23,202(74),182(18),172(31), 150(14),140(19),130(22),115(21),103(10),85(22),73(100).-High resolutions:

4. A process for the preparation of a compound according to any one of claims 1 to 3 which comprises - culturing Polyangium DSM 6267 in a medium containing carbon sources, nitrogen sources and mineral salts, - extracting the harvest cell mass with acetone, - concentrating the (combined) extract, - taking up the concentrate in ether/water, - separating off the aqueous phase and if necessary extracting with ether, - concentrating the (combined) ethereal phase, WO 92/11258 PCT/EP91/023??

- taking up the concentrate in methanol/heptane, - separating off the methanol phase and if necessary extracting with heptane, - then concentrating and extracting with tert-butyl methyl ether, - filtering the extract in tert-butyl methyl ether over Florisil, - crystallising thiangazole (or antibiotic and/or fungicidal activity) from the extract and - if appropriate converting the thiangazole (or antibiotic and/or fungicidal activity) into a therapeutically acceptable acid addition salt.

5. A process for the preparation of cis-trans isomers of a compound of formula I according to claim 1 which comprises subjecting the compound of formula Ia according to claim 2 to treatment with UV light and separating the resulting isomers by chromatography, if appropriate isolating those isomers and if appropriate converting them into pharmaceut-ically acceptable acid addition salts.

6. A pharmaceutical composition (with the exception of those for controlling viral diseases) consisting of a compound according to any one of claims 1 to 3 or comprising one of those compounds or one of the physiologically tolerable acid addition salts thereof and at least one customary formulation adjuvant.

7. A pharmaceutical composition according to claim 6 for controlling Arthropoda or Acarina, and also for controlling parasitic diseases in humans or animals (with the exception of those for controlling viral diseases).

8. A pharmaceutical composition according to claim 7 for controlling helminths.

9. A compound according to any one of claims 1 to 3 for use in a method for the thera-peutic treatment of the human or animal body (with the exception of those for controlling viral diseases).

10. A pesticide for protecting plants for agriculture, forestry and/or horticulture, based on a compound according to any one of claims 1 to 3 or an acid addition salt thereof together with a customary carrier.

11. The use of a compound according to any one of claims 1 to 3 for the preparation of a pharmaceutical composition, a pesticide or a crop-protection composition.

12. The use of a compound according to any one of claims 1 to 3 for the preparation of a composition against Arthropoda, preferably insects or pests from the class of the Acarina.