Derivatives marcfortine/paraphernalia, the method of suppressing pests, insects or nematodes strain of the fungus

 

(57) Abstract:

Disclosed 18-timecharter derived natural products marcfortine a, b and C, C-13 thiomargarita and their derivatives, the new N-1 marcfortine a, b and C and their derivatives, a new N-1 parsername and its derivatives, useful in the treatment and prevention of infections caused by helminths and arthropods in animals and plants. Synthetic derivatives have the formula 1. 3 S. and 4 C. p. F.-ly, 2 tab. .

Marcfortine are known compounds and are described Polonsky et al. B. Journal of the Cemical Society Chemical Communications, 1980, 601-602 (Marketin A) and Tetrahedron Letters, 1981, 22 1977-1980 (Marcfortine B and C). Connections are fungal metabolites of Penicillium rogneforti. Macrophomina structurally related to paraphernalia, which are also known compounds Paraphernalia disclosed Yamazaki at al Tefraheolron Letters 1981 22 135-136, and Blanchflower et.al. Journal of Antibiotirs, 1991, 44, 492-497. U.S. patent 4 866 060 and 4 923 867 describe the application of marcfortine A, B and C and several of their derivatives, as suitable for the treatment and prevention of diseases caused by parasites in animals.

Paraphernalia has the following structure:

< / BR>
Markvorsen A has the following structure:

< / BR>
Marcfortine B has the following structure is e derivatives marcfortine and paraphernalia and (12) oxides as well as getting VW 29919 (parsername) and VM 55596 (N(12) oxide paraphernalia) (among other things) from Penicillinm Sp. IMI 332995.

U.S. patent 4 873 247 describes derivatives paraphernalia and strain Penicillinm charlessi MF 5123 (ATCC 20841) to obtain paraphernalia. U.S. patent 4 978 656 (as well as EP 390532-A, EP-301742-(A) describes the various synthetic derivatives, as well as getting paraphernalia of Penicilli um charlelli MF 5123 (ATCC 20841).

Paraphernalia is a compound produced under certain conditions fungal organism Penicillium paraherguei. WO 92/00300 (publ. January 9, 1992) discloses the synthesis of compounds related to paraphernalia received from (produced) Penicillium paraherguei deposited in the C. A. C. International Mycological Institute, Ferry Lane, Kew, London number CMI 68220.

This invention relates to the synthesis of C-18 tomarketing A, B and C and their derivatives, C-18 thiomargarita and its derivatives, the new N - I marcfortine, A, B and C and their derivatives, a new N - I paraphernalia and its derivatives, and the use of these compounds as proliferating agents.

The purpose of the invention consists in the description of these derivatives timecharter, thiomargarita, marcfortine and paraphernalia.

In addition, the purpose of this invention is the method of obtaining these compounds.

A further objective is to describe the by parasites.

In addition, a further purpose is to describe compositions for the treatment of diseases caused by parasites containing the new compounds of this invention as their active ingredient.

The compounds of this invention represented by formula I:

< / BR>
where

m is 0 or 1;

n = 1;

W = 5 or 0

R14aor R14bis a hydrogen atom;

R24and R25form a double bond;

R18ais a hydrogen atom or a C1-C7- alkyl;

R1is a hydrogen atom;

-C(O)R1group (where R1- cyclo C3-C8- alkyl, phenoxycarbonyl, -C(O)-C2-C24-alkyl, nitrophenoxyacetic, 1,3-dioxo-2H-isoindolyl, dinitrobenzenesulfonyl, trichloromethylsulfuryl, C1-C4-alkoxycarbonylmethyl, benzazolyl, tetrahydropyranyl, 1,4-dioxy-C2-C7alkenyl

or R1group-C(O)-NR4R5where R4and R5together with the nitrogen atom form a saturated heterocyclic ring: pyrrolidinyl, morpholinyl, piperazinil, unsubstituted or substituted C1-C4-alkyl, C1-C4-alkanoyl, C1C4-alkoxycarbonyl, 1,3 - benzodioxolyl-C1-C4-alkyl, pyridinyl, haloge the>alkoxycarbonyl, trifter C1-C4-alkyldiethanolamine, or R4and R5form piperidinyl, unsubstituted or substituted phenyl or mono - or di-substituted C1-C4-alkyl,

or their pharmaceutically acceptable salts, or hydrates.

Another aspect of the present invention is an 18-timecertain or derivatives of the formula II

< / BR>
or N-1-marcfortine IV

< / BR>
where the Deputy shall have the same meaning as for formula I.

Preferred compounds I where R1- and - rich heterocycle.

Examples of the heterocycles formed-NR4R5that are

4-morpholine,

4-phenyl-1-piperazine,

4-(2-pyridinyl)-1-piperazine,

2,6-dimethyl-4-morpholine,

1-pyrrolidin,

4-methyl-1-piperazine,

1-piperidine,

4-phenyl-1-piperidine,

2 methylpiperidin,

3 methylpiperidin,

4 methylpiperidin,

N-methylpiperazin,

1-methylhomopiperazine,

1-acetylpiperidine

Preferred compounds of the invention are

18-timecharter A (compound 2),

18-timecharter B (compound 2A),

1-pyrrolidinyl-marcfortine A (compound 12),

1-cyclopropanecarbonyl-marcfortine A (compound(4-nitrophenoxyacetic)-marcfortine A (compound 12D),

1-(1-piperidinylcarbonyl)-marcfortine A (compound 12F),

1-[[4-(etoxycarbonyl)piperazine-1-yl] carbonyl]-marcfortine A (compound 12G),

1-[[4-(benzyl)piperazine-1-yl]carbonyl]-marcfortine A (connection 12H),

1-[[4-(1,3-benzodioxol-5-ylmethyl)piperazine-1-yl] carbonyl]- -marcfortine A (compound 12I),

1-[[4-(methyl)piperazine-1-yl]carbonyl]-marcfortine A (compound 12J),

1-(1,3-dihydro-1,3-dioxo-2H-isoindole-2-yl)-marcfortine A (connection 12K),

1-[[4-(pyridin-2-yl)piperazine-1-yl] carbonyl] -marcfortine A (connection 12L),

1-[[4-(phenyl)piperazine-1-yl]carbonyl]-marcfortine A (connection 12M),

1-[[4-(chlorocarbonyl)piperazine-1-yl] carbonyl] -marcfortine A (connection 12N),

10,10"-(1,4-decarbonylation)bis(6', 7',8',9',10',10'a - hexahydro-1', 1', 4,4,12'-pentamethyl)-[2'S-[2 alpha, 3 a.alpha, 9 a.alpha, 10/2"'R*, 3"'aS*9"'aS, 10"'aR*)10'a.beta]]-Spiro[4H,8H-[1,4]dioxano[2,3-g]indole - 8,2'(3'H)-[1H,4H-3a,9a](aminomethane of cyclopent[b]hemolysin]-9,11'(10H)-dione (compound 120),

1-[[4-(phenyl)piperidine-1-yl]carbonyl]-marcfortine A (compound 12P),

1-[[4-(dimethyl)piperidine-1-yl]carbonyl]-marcfortine A (connection 12Q),

1-[[4-(5-chloropyridin-3-yl)piperazine-1-yl] carbonyl] -marcfortine A (connection 12R),

1-[[4-(4-chlorophenyl)piperazine-1-yl] carbonyl] -marcfortine A (with. 9'a.alpha, 10(2"'R*, 3"'aS*, 9"'aS*, 10"'aR*) 10'a, beta]]-Spiro[4H, 8H-[1,4]dioxano[2,3-g] indole-8,2'(3'H)- [1H, 4H-3a, 9a] (aminomethane)of cyclopent[b] hemolysin] -9,11'(10H)-dione (compound 12T),

1-[[4-(Trichloroisocyanuric)piperazine-1-yl] carbonyl] - marcfortine A (compound 12U),

1-[[4-(3-trifluromethyl-thiadiazole-1-yl)piperazine-1-yl] carbonyl] - marcfortine A (connection 12V),

1-acetoxymethyl-marcfortine A (compound 13)

1-(2,4-dinitrobenzenesulfonyl)-marcfortine A (compound 14),

1-(4-morpholinomethyl)-marcfortine A (compound 14A),

1-(trichloronitromethane)-marcfortine A (compound 14B),

1-(methoxycarbonylmethyl)-marcfortine A (compound 14C),

1-(benzazolyl)-marcfortine A (compound 14D),

1-(4-morpholinylcarbonyl)-marcfortine AN-oxide (compound 15),

1-(2-tetrahydrofuranyl)-marcfortine A (compound 16).

Pharmaceutically acceptable salts means salts that are useful for the purpose of the compounds of this invention as medicaments, these salts include mesilate, hydrochloride, hydrobromide, hydroiodide, sulfate, phosphate, acetate, propionate, lactate, maleate, malate, succinate, tartrate, and the like. These salts may be in hydrated form.

The compound of formula II preloadnetthing their substituted C-24, C-25, N-1 and N-18a derivatives pentasulfide phosphorus or preferably 2,4-bis(methoxyphenyl)-1,3-dithia-2,4-diphosphate-2,4-disulfide (reagent Lawesson'a) selectively gives the corresponding 18-thio-derivatives. Suitable C-24, C-25, N-1 and N-18a substituted derivatives marcfortine A, B, and C for this reaction is easily prepared by the methods described in [1], the description of which is incorporated herein by reference. The reaction is conducted in a suitable inert solvent, such as pyridine, kallidin, toluene (preferably), xylene, dioxane, tetrahydrofuran and the like, at temperatures of 10 - 180oC, preferably 80 to 140oC. Or C-24, C-25 and N-1 derivatives can be prepared from 18-tomarketing. For example, a large series of analogues 18-timecharter can be prepared by alkylation or acylation of the N-1 and N-18a substituted 18-tomarketing. Such derivatives can be easily prepared by the sequential processing solution 18-timecharter A, N-18a-substituted 18-timecharter B or N-18a-substituted 18-timecharter C in proton organic solvent, such as tetrahydrofuran, diethyl ether, benzene and the like, an excess of a strong base such as potassium hydride (preferred), sodium hydride, stage agent at temperatures in the range of 0 - 50oC from 0.25 to 48 hours

Suitable alkylating agents include alkyl bromides, alkyl iodides, alkyl sulfonates, alkenyl the iodides, quinil bromides, alkoxyalkyl chloride and the like. Suitable alleluya agents include acyl anhydrides, acyl chlorides, acyl bromides, substituted benzazolyl chlorides, substituted benzosulfimide anhydrides, sulfenyl chlorides, isocyanates, carbarnoyl chlorides, chloroformate and the like.

Additional series of derivatives can be produced by modification of C24-C25 double bond 18-tomarketing A, B and C. 24, 25 dihydro analogues easily prepared by mixing a solution of a suitable 18-tomarketing in an alcohol solvent such as methanol, ethanol, propanol and the like with a catalyst, such as palladium, platinum, Tris (triphenylphosphine) - chloride and the like in the presence of gaseous hydrogen. The product, which is similar to 24,25-dihydro-18-marcfortine, can be isolated and purified using methods known to a person skilled in this technique. It should be noted that the above reactions for the modification of another part of the 18-timecharter patterns can also be applied to 24, 25-dihydro 18-timecharter analogs is due analogues 18-tomarketing can be obtained through 24, 25 dibromide, which is easily prepared by treatment of a solution of 18-tomarketing in halogenosilanes a solvent such as dichloromethane, chloroform, chetyrehhloristy carbon and the like I molar equivalent of bromine at a temperature range of -20 to 25oC for a time of from 0.25 to 8 o'clock This method gives the corresponding derivative of 24.25-dibromo 24,25-dihydro-18-tomarketing, which can be isolated and purified by using techniques known to experts in this technique.

It should be noted that 24,25, sodium dichloro-analogue can be prepared by substitution of bromine for chlorine in the above-described method. Analogues of 24.25 - dibromo 24,25-dihydro-18-marcfortine described above, are used as intermediates for additional derivatives. Thus, treatment of a solution of the dibromide in an alcohol solvent such as methanol, ethanol, propanol and the like, a strong base, such as 1,8 diazabicyclo [5.4.0] undec-7-ene (DWI)in the temperature range 0 - 30oC for 0.25 to 24 h gives analogues 24-alkoxy-25 bromo-24, 25 dihydro-18-tomarketing, which can be isolated and purified by using techniques known to experts in this technique. These 24 alkoxy, 25-bromo derivatives of benzene, toluene, hexane and the like, regenerating agent-based tin hydride, such as tri-butyl tin hydride, three-pencil tin hydride and the like with or without adding a radical initiator, such as azobis-isobutyronitrile (AIBN) in the temperature range 25 - 120oC for 0.5 - 48 hours This method gives the corresponding derivatives 24-alkoxy-18-timecharter (R24= lower alkoxy in the overall structure), which can be isolated and purified by using techniques known to experts in this technique.

The main ways to get heteroaromatic N - oxides can be found in Chapter II of "Chemistry of the Heterocyclic N-Dxides" A. R. Kattitzky and T. M. Ladowski, pyblished 1971 Academic Press (Vol.19 of ORGANIC CHEMISTRY - A Series of Monographs).

Typically the N - oxide is formed by the interaction and nagarbhavi acid in an aprotic solvent. It is most convenient to use aromatic nagkalat in nonpolar solvent as the reaction can usually be carried out at room temperature. Suitable aromatic nagkalat include adventurou acid, chloronitrobenzene acid and tereftalevuyu acid.

The compound of formula IV is prepared by alkylation or acylation of the N-1 markf the processing solution marcfortine A or N-18a substituted marcfortine B and C in an aprotic organic solvent, such as, tetrahydrofuran, diethyl ether, benzene and the like with an excess of a strong base such as potassium hydride (preferred), sodium hydride, utility, tert.-butyl potassium, and the like, followed by treatment with a suitable alkiliruushim or allermuir agent in the temperature range 0-50oC for 0.25 to 48 hours Suitable alkylating or alleluya agents include alkanoyloxy bromides, aminosulfonyl, phosphoryl chlorides, phosphonyl chlorides, acyl anhydrides, acyl chlorides, acyl bromides, substituted benzazolyl chlorides, carbarnoyl chlorides, substituted penicilliformis and the like. Processing marcfortine A and N-18a substituted marcfortine B and C and C24-C25 modified marcfortine under these conditions gives 1-N-selesnya analogues, which can be isolated and purified by using techniques known to experts in this technique.

Preparation of starting material

Paraaramid isolated from Penicillium Sp. IMI 332995 and/or Penicillium charlessi MG 5123 (ATSS 20841) using standard enzymatic and separation techniques. The selection is described in detail in U.S. patent 4973247 and 4978656 and EP 390532-A, EP-301742-A and WD91/09961 (all included here as a reference).

is presented in [1].

Marcfortine A, B, and C are selected, along with the previously known roquefortine as fungal metabolites Anicillinm roguerforti using standard enzymatic and separation technique. The selection, as well as analytical and structural characteristics marcfortine A, described in detail in Polonsky et al Journal of the Chemical Sosiiety Chemical Communisations 1980, 601-602. The selection, as well as analytical and structural characteristics of marcfortine B and C, described in detail in Polonsky et al Ketrahedron Letters 1981, 22, 1977-1980.

An alternative and more preferably, Marcfortine A and C can be isolated from Penicillium sp. IP 7780 (number of strains in culture Collections of IP 7780, The decision Upjohn Company, Kalamazoo, MI). This strain was isolated from soil sample collected in Illinois, deposited in the U.S. Department of Alricnetnre patent cnltnre collection in Peoria, IL and catalogued under the number NRRL 18887. For further characterization of the fungus was carried out taxonomic research according to the methods and materials described John I. Pitt, The Genus Pinicillium, Acodemic Press, L ondon, (1979). Disputes and surface hyphae were investigated by scanning electron microscopy according to methods Dietz, A. and Matthews, J. Appl. Microlilogy 18: 694-696 (1969). Intact conidiophore was observed visually using light microscopy [A. H. S. Onions et al, Smith's Introd lannou a Petri dish, containing glass beads, microscopic preparation and coating glass sterilized. A small chunk of potato agar with dextrose placed on a microscopic preparation and inoculant on all four sides fungal structure. Cover glass is placed on the inoculated block of agar and add sterile water to maintain humidity. The camera incubated for six days at the 24oC. Microscopic preparation is prepared by removing the top glass and placing it on a drop lactophenol cotton sineni.

Characteristics of the strain Penicillium sp. IP 7780 (NRRL 18887) following.

Morphology vimutta brush (two branch point between conidia and leg). These branching (Metula) support bearing structure of Fieldy and conidia. Conidiophore (approximately 35 microns) end in whorls of 2-5 (10-14 μm) of metal. Fieldy was in the form of cones (as Greek wine vessel) in whorls from 2-5 (7 μm). Conidia were smooth and spheroidal (2 μm), usually forming long ginesthai. Wall legs were smooth.

The culture was inoculable on three Petri dishes on yeast agar of čapek (CYA), of which one was 6 cm in diameter, the other contained agar malt extract (which 0.05% tween 80). Inoculum loop with conidia introduced into a test tube and mixed. The loop with the suspension was inoculable in the sample in three places on each Cup. For the introduction of inoculum in 6 cm Cup was used needle. Used incubation mode: one CYA Cup plus MEA and 25 G N Cup at the 24oC, one CYA Cup at 37oC and 6 cm CYA Cup at the 5oC. After seven days, diameters, colors, and other characteristics of the colonies were recorded and placed next to the table. 1. On potato dextrose PDA, Difco) agar at the bottom or reverse side of the colony appears dark red.

The sexual stage has not been observed. These results are in culture (NRRL 18887) was determined by Penicillium. The identifier for identifying subcultures. On Penicillium provider, only penitsillinovy type determines the subgenus to which it relates. This species has some characteristics that distinguish it from subgenus Biverticillium. For example, the brush are whorled. View reliably produces colonies of more than 10 mm in diameter for 7 days on agar with glycerol nitrate. Metulla is longer than Fieldy and in whorls from 2-5. These characteristics allow us to refer this strain Penicillium sp. (NRRL 18887) to the subgenus Furcatum.

The above description Il is-breaking. In addition, this invention also includes mutants of the above-described microorganism. For example, those mutants that obtained by natural selection, or the mutants, obtained by mutating agents, including ionizing radiation, such as ultraviolet radiation or chemical mutagens, such as nitrosoguanidine, or similar treatments, are also included in the purpose of this invention.

The object and purpose of this invention is the inclusion of mutual or internal genetic recombination processes using genetic techniques, well known to experts in the art, such as, for example, algae, translite (transultion) and methods of genetic engineering.

Penicillium sp. IP 7780 (NRRL 18887) can be cultured under aerobic conditions by a method that is usually applied in the technique of cultivation known strains of the genus Penicillium.

As components of the environment can be used any well-known nutrients for Penicillium. For example, as assimilated carbon source, glucose, glycerol, maltose, dextrin, starch, lactose, sucrose, molasses, soybean oil, cottonseed oil, etc. oeva flour, peanut meal, cotton seed meal, fish meal, liquid corn impregnation, peptone, rice bran, meat extract, yeast, yeast extract, sodium nitrate, ammonium nitrate, ammonium sulfate, etc. can be used. And inorganic salts such as sodium chloride, phosphates of sodium, calcium carbonate, etc. can be added to the environment for crops. If necessary, can also be added minor amounts of a metal salt. In addition, minor amounts of heavy metals can be added if necessary.

In particular, in the cultivation of Penicillium sp. (NRRL 19997) under aerobic conditions with success can be used conventional methods of aerobic cultivation, such as, for example, growing on dry residue, cultivation under aeration and stirring (agitation), cultivation with shaking, etc. To enzymatic environment must be added to the trace metals, such as zinc, magnesium, manganese, cobalt, iron and the like, as must be supported by the level of reverse-osmosis water.

In carrying out cultivation under aeration and agitation can be successfully applied antifoaming agents such as silicone oil, vegetable oils, surfactants, etc.

s cultivation can usually be 20-30oC, particularly preferred about 21oC.

Cultivation may continue up until Marcfortine A significantly accumulate in the environment, culture, usually from 20 to 240 hours, preferably 48 to 168 h, and after cultivation Marcfortine A can be isolated and separated from the liquid medium for cultures (culture broth) by a suitable combination of different methods. For example, it may be extraction with an organic solvent, for example diethyl ether, ethyl acetate or chloroform; dissolution in the more polar solvent, such as acetone or alcohol; removing impurities less solvent such as petroleum ether or hexane, absorption chromatography on activated carbon or silica gel; gel filtration through a column of "Sephadex" (supplied by Pharmacia Co., Ltd, USA), and the like.

Marcfortine B can be prepared from Marcfortine A by biotransformation using a microorganism in water nutrient medium containing assimilated nitrogen source under aerobic conditions.

This invention also provides a biologically pure culture (organisms obtained from culture collections, as clearly exposed organisms) kind Sipha members of the genus Cunnirghamella, characterized as a species selected from the group consisting of NRRK 1368, 1393 or 3655 and ATCC a to obtain Marcfortine B, which involves attaching Makhorin A metabolisable the cultures of these microorganisms.

By "biotransformation" means the use of microorganisms or the allocation of partially purified enzymes for the conversion of this substrate into a useful product. N. With. Davies, et al, "Biotransformations in Dreparative Organic Chemistrg", Academic Press, N. Y., 1989, p.IX.

By "b" means the carrying out of the processes of metabolism. Definition of metabolism can be found in A. L. Lehninger, "Principlles of Biochemistry", Worth Pnbl., New York, 1982, p.333.

By using the new method of this invention Marcfortine B is obtained by adding Marcfortine A growing cultures of these organisms. Genus Cunnirghamella is preferred to perform this transformation, including Cunnirghamella echinulata sublp. elegans (-) NRRL 13b8, Cunninghamella blakesleeana (+) ATTC 8b88a Cunnirghamella echinufata subsp. elegans NRRL 1393, Cunnirghamella echinulata NRRL 3655;

in particular strain NRRL 3655 is preferred.

Podculture Cunnirghamella echinulata subsp. elegans (-) was taken on Deposit Budapest Treaty in the permanent collection of the Northem Region Research Center, ARS; U. S. Dept of Agriculture; Peoria, Illinois, USA.

She was assigned a room NRRulture Collection, Rockville, MD, USA.

It was assigned the number ATCC 8688b.

Podculture Cunnirghamella blakesleeana (+) taken on the Deposit of the Budapest Treaty in the permanent collection of the American Tupe Culture Collection, Rockville, MD, USA.

It was assigned the number ATCC 8688a.

Podculture Cunnirghamella echinulata subsp. elegans (-) taken on the Deposit of the Budapest Treaty in the permanent collection of the Northem Region Research Center, ARS; U. S. Dept of Agriculture; Peoria, Illinois, USA.

Its serial number is NRRL 1393. Podculture Cunnirghamella echinulata subsp. elegans (-) taken on the Deposit of the Budapest Treaty in the permanent collection of the American Tupe Culture Collection, Rockville, MD, USA.

She assigned a number ATCC 10028b.

Podculture Cunnirghamella echinulata was taken on the Deposit of the Budapest Treaty in the permanent collection of the Nocthern Region Research Center, ARS; U. S. Dept of Agriculture; Peoria, Illinois, USA.

She assigned a number NRRL 3655.

The use of these microorganisms in the method of the present invention is expressed in obtaining Marcfortine B from Marcfortine A.

Marcfortine B obtained when Cunnirghamella echinulata subsp. elegans (-) NRRL 13b8, Cunnirghamella blakeslleana (+) ATTC 8b88a Cunnirghamella echinulata subsp. elegans NRRL 1393, Cunnirghamella echinulata NRRL 3b55

fermented in an aqueous nutrient medium under aerobic conditions dives in the presence Marcfortine A. Typically, the microorganism is fermented in a nutrient medium containing a source glycerin, starch, corn starch, lactose, dextrin, molasses and the like. Preferred nitrogen sources include cottonseed flour (flour cottonseed), liquid corn impregnation, yeast, avtorizirovannye brewer's yeast with a dry matter of milk, soy flour, corn flour, dry matter of milk, casein is digested by pancreatic enzyme extract peptone animal origin, extracts of meat and bones, and the like. Can be mostly used a combination of these sources of carbon and nitrogen. There is no need to add traces of metals such as zinc, magnesium, manganese, cobalt, iron and the like, because the quality of the components used water from the tap and unrefined ingredients.

Production Marcfortine B can be induced at a certain temperature conducive to satisfactory growth of the microorganism approximately between 23 and 32oC and preferably at about 28oC.

Usually the best education Marcfortine B is approximately between 1 to 4 days after adding Marcfortine to A growing culture, and preferably for a period of about 2 days. Fermentative broth usually remains from outstay and in part on the initial pH of the medium for crops. It is advantageous to adjust the pH to about 6.5-7.5 and preferably to 7.2 before sterilization.

When growth is carried out in shake flask or large vessel and collections, it is preferable to use the vegetative form of the organism for inoculation, instead of using a form of dispute to avoid a pronounced delay in the generation of Marcfortine B and the attendant inefficient utilization of the equipment. Thus, it is desirable to obtain a vegetative inoculum in an aqueous nutrient medium by insulinopenia this environment aliquot breakdown of soil or culture on the sloped agar. When a young, active vegetative planting material thus selected, it is transferred aseptically to other shake tubes or collections. The environment in which the vegetative inoculum receive may be the same or different from that used to obtain Marcfortine B, the length of the process receive adequate growth of the microorganism.

Can be used various methods of isolation and purification Marcfortine from A fermentation broth, for example by the method of absorption chromatography with subsequent alyazia suitable solvent, speaker chromatogr

The preferred method of selection Marcfortine B is the extraction of the (raw) whole beer. Methods column chromatography, preferably on silica gel, is used to perform the initial cleaning. Final cleaning Marcfortine B is achieved by chromatography and crystallization from organic solvents.

The method further illustrates A method of biotransformation of the invention.

These compounds of this invention are unexpectedly highly active antiparasitic agents against endo and ecto parasites, particularly helminths (parasitic worms) and arthropods that cause numerous diseases caused by parasites in humans, animals and plants.

Diseases caused by parasites, can be caused by endoparasites and ectoparasites. Endoparasites are parasites that live inside the host's body or inside the body (such as the stomach, lung, heart, intestines, and so on) or just under the skin. Ectoparasites are parasites that live on the outer surface of the host, but still pull nutrients from the host.

Disease caused by endoparasites, usually called the Worms are widespread and serious global economic problem, associated with infection of domestic animals, such as pigs, horses, sheep, cattle, goats, dogs, cats and poultry. Many of these infections are caused by a group of worms called nematodes, which cause diseases of different organs in animals worldwide. These diseases are often serious and can lead to death of the infected animal. The most common genera of nematodes infecting the above-mentioned animals, include At, Trichostrongylus, Ostertagia, Nematodirus, Cooperia, Ascaris, Bunostomum, the recommended dose rate, Chabertia, Trichuris, Strongylus, Trichonema, Dictyocaulus, Capillaria, Heterakis, Toxocara, Ascaridia, Oxyuris, Ancylostoma, Uncinaria, Tokascaris, and Parascaris.

Many parasites are species-specific (infect only one host) and the majority also has a preferred site of infection in animals. So, At the and Ostertagia mainly infect the stomach, whereas Nematodirus and Cooperia mainly attack the intestines. Other parasites prefer to linger in the heart, eyes, lungs, blood vessels and the like, while others are fixed subcutaneous parasites. The worms can cause weakness, weight loss, anemia, damage to the bowel, lack of power and damage to other organs. If this disease is left without Le is issued, such as ticks, mites, lice, autumn of Zhigalko, jagalchi cow small, patelnie flies, fleas, etc., is also a serious problem. Infection with these parasites is expressed in blood loss, skin damage, can interfere with normal digestion and lead to weight loss. These infections can also lead to the development of serious diseases such as encephalitis, anaplasmosis, smallpox in pigs, etc. that can lead to fatal outcome.

Animals can be infected by several species of parasites at the same time as infection by one parasite can weaken the animal and make it more susceptible to infection with a second type of parasites. Thus the connection with a wide range of activity has advantages in the treatment of such diseases. The compounds of this invention have unexpectedly high activity against these parasites, and in addition are also active against Dirofilaria in dogs, Nematospiroides and Suphacia in rodents, stinging insects and migratory licino dipterans, such as Hypodorma sp. in cattle and Gastrophilus in horses.

These compounds are also useful against endo and ecto parasites that cause diseases caused couples is Rasimov of the genus Ancylostoma, Necator, Ascaris, Strongyloides, Trichinella, Capillaria, Trichuris, Enterobius, etc. Other endoparasites, which infect humans, are found in the blood and other organs. Examples of such parasites are thread worms Wucheria, Brugia, Onchocerca and the like, as well as extraintestinal phase of intestinal worms Strongylides and Trichinella. It's the parasites that parasitize on the person, include arthropods, such as ticks, fleas, mites, lice and the like and as with domestic animals, infection with these parasites can result in development of serious and even fatal diseases. These compounds are active against these endo and ecto parasites and, in addition, also effective against biting insects and other dipteran parasites that bothers a person. Current connections when used orally or parenterally, are prescribed in doses in the range of about 0.05 to 20 mg/kg body weight of the animal. These compounds are also useful against the occupants of pests, such as Blatella sp (cockroaches), Tineola sp (a mol), Attagenus sp. (koreeda), Musca domestira (room fly) and against Solenopsis Invicta (ant Richter).

In addition, the compounds are useful against agricultural pests, such as aphids (Acyrthiosiphon sp.), locusts and cotton dolmeh on uncompleted stages of development, living in plant tissue. The compounds are also useful as nematocide for control of nematodes in the soil, which can be important for agriculture. When used as antiparasitic agents for animals, the compounds of this invention can be assigned internally or orally, or by injection, either locally in the form of liquid irrigation or as a shampoo.

For oral administration the compounds can be assigned in the form of capsules, tablets, balls for infusion of medication or otherwise, they can be mixed with food for the animals. Capsules, tablets and balls contain the active ingredient in combination with an appropriate binder carrier, such as starch, talc, magnesium stearate, or dicalcium phosphate. These unit dosage forms are prepared by the close of mixing the active ingredients with suitable solvents, fillers, desintererada agents, suspendida agents and/or binders, so as to achieve a homogeneous solution or suspension. Inert ingredients are those that will not interact with these compounds and which are not toxic for animals to be treated. Suitable inert ingredi can contain a wide variety of active and inactive ingredients, depending on numerous factors such as the size and type of animal and the type and severity of infection. The active ingredients can also be administered in the form of food supplements by simply mixing the compound with food or applying the compounds to the surface of food. Otherwise, the active ingredient may be mixed with the inert carrier and the resulting composition may then be or mixed with food or directly fed to the animal. Suitable inert carriers include corn meal, citrus meal, enzyme residues, soy flour, dry cereals, etc., the Active ingredients are intimately mixed with these inert carriers by milling, stirring, crushing or hallowene, so that the final composition contains 0.001 to 5.0 wt.% the active ingredient.

Otherwise the connection can be assigned parenterale through injection of a composition containing the active ingredient, dissolved in an inert liquid medium. Injection can be performed intramuscularly, intraruminal, vnutritrahealno or subcutaneously. Formulations for injection contain the active ingredient mixed with a suitable inert liquid medium. Acceptable liquid carriers include historical solvents, such as solketal, glycerol formal and the like. Alternatively, you can also use aqueous parenteral formulations. Vegetable oils are dissolving or suspendirovanie active ingredient in a liquid medium so that the final composition contains from 0.005 to 20 wt.% the active ingredient.

Topical application of these compounds is the possibility of using liquid sprinklers or shampoos containing the current connection in the form of aqueous solutions or suspensions. These formulations usually contain suspendisse agent, such as bentonite, and usually should also contain antifoaming agent. Acceptable are compounds containing from 0.005 to 20 wt.% these compounds. These compounds are mainly useful as antiparasitic agents for the treatment and/or prevention of helminthiasis in domestic animals such as cattle, sheep, horses, dogs, goats, pigs and poultry. They are also useful in the prevention and treatment of infections from parasites, caused by parasites in these animals, such as ticks, mites, lice, Zhigalko and the like. They are also effective in the treatment of infections caused by parasites in humans. In such treatment and the other not related antiparasitic agents. Doses of these compounds necessary for the best results depend on several factors such as the type and size of the animal, the type and severity of infection, treatment and used for the connection. Oral treatment with these compounds at doses of 0.005-50 mg per 1 kg body weight of the animal, either in a single dose or in multiple doses, held in a few days, gives good results. A single dose of one of these compounds usually gives excellent control, however, repeated doses may be given to prevent re-infection or parasites, which occasionally remain. The technique of treatment of these animals are known compounds for professionals working in the veterinary field.

The compounds of this invention can also be used to combat agricultural pests that destroy crops or fields, or during storage. Compounds used for such purposes in the form of liquids for spraying, dust products, emulsions and the like or for growing plants or harvested crop. Methods of using these compounds for such purposes are known for working in the field of agriculture experts the doctrine and selection Marcfortine A

The method of fermentation seeds

Seed fermentati inoculant using agar plugs culture of Penicillium sp. IP 7780 (NRRL 18887), stored in liquid nitrogen. Three tubes are thawed and used as inoculum. GS - 7 are from glucose and cotton flour (mark "Pharmamedia" "by Traders Protein, Procterk Gamble Oilseed Products Co., Memphis, TN, U. S. A.

Use tap water without additional purification for the hydration of the components and pH of the medium was adjusted to 7.2 with NH4OH. The medium was dispensed in not containing walls of the flask, representing a closed system, 300 ml to 1000 ml flask and sterilized by autoclaving at 121oC for 30 minutes Each flask with a closed system containing 300 ml GS - 7 environment, inoculable three agar plugs culture of Penicillium sp. 7780 (NRRL 18887) and were shaken in a rotary vibrator at 250 rpm for 36 h at 22oC.

Method of secondary fermentation seeds

Culture ripe seeds are used as inoculum for secondary environment at 0.3% of the dose administration. The secondary environment is composed of glucose monohydrate (brand Cerelose by C. P. C. International) 25 g flour cottonseed (mark "Pharmamedia") 25 g MgCl26H2O USD 329.8 mg; MnSO4H2O 11.4 mg; FeSO47H2O 3.2 mg; B>6H2O 0.1 mg; CuSO45H2O 3.1 mg and antifoaming silicone (brand SAC - 471 Antifoam) 0.5 ml per 1 liter of water to provide reverse osmosis. Components sufficient for 200 l secondary environment for seeds hydratious in water with indicator reverse osmosis to q. s. volume of 190 l 250-liter fermenter. After establishing the pH of the medium was adjusted to 7.2 with NH4OH and then the medium is sterilized at 121oC for 30 minutes Two flasks closed system culture ripe initial seeds used as inoculum at 0.3% of the dose administration. Secondary seed culture incubated at 22oC 125 slm aeration when the back pressure (0.35 kg/cm2and 250 rpm for 36 hours

The method of carrying out fermentation

The production environment consists of beet molasses 50 g of fish meal (brand Henhaden Select Fish Meal) 16 g, yeast extract (brand Fidro) 10 g; MgCl26H2O USD 329.8 mg; MnSO4H2O 11.4 mg; FeSO47H2O 3,29 mg; Na2MoO42H2O 1.8 mg; CaCl22H2O 367,6 mg; NaCl 84,2 mg; KCl to 5.8 mg; ZnSO47H2O 0.1 mg; CoCl26H2O 0.1 mg CuSO45H2O 3.1 mg and antifoaming silicone (brand SAG - 471) 0.5 ml per 1 l of water with indicator reverse osmosis.

The components of the environment, sufficient DL is ing the pH of the medium was adjusted to 7.0 with KOH, and then the medium is sterilized at 123oC for 30 minutes a Culture ripe secondary seed is used as inoculum at 1.0% of the dose administration. The culture is incubated at 22oC 2,500 slm aeration at 5 psig back pressure and 250 rpm for 96 hours

Selection marcfortine A

4900 l fermented volume of harvested cells, grown in culture, passing through finely chopping mixer in the vessel for collecting cells. After migration, add 4% wt/about diatomaceous earth and 1/2 volume of methylene chloride. The solution collected cells are then filtered using the press for filtering. The filter cake is washed with 2 times 10% by volume of methylene chloride.

The obtained filtrate is decanted to remove the aqueous phase. The remaining phase methylene chloride-enriched product, then concentrate to a volume of 44 liters of Concentrate is then brought to the desired condition using 20% of the concentrate volume (9 l) methylene chloride and diatomaceous earth on the filter.

53 l squared away concentrate is then clear for the Department Marcfortine A from other components using chromatography on silica gel and crystallization.

Before chromatography squared away concentrate is divided into four diameter 9", filled with 25 kg of dry silica gel (volume of filler 59 l). The loaded column elute 120 l of 10% acetone in methylene chloride, 120 l 20% acetone in methylene chloride, 120 l of 30% acetone in methylene chloride, 160 liters of 40% acetone in methylene chloride and 130 liters of acetone, collecting 30 and 40% eluate fractions 20 HP Eluate analyzed by TLC, using for example a solvent system composed of 6% isopropanol and 0.3% ammonium hydroxide in methylene chloride, for the manifestation of plates of silica gel.

Faction Marcfortine A (containing a small amount Marcfortine A chromatographic which goes together with A) crystallized from acetone. The appropriate fractions (40-100 l) concentrate under reduced pressure to approximately 5 liters of Solution (or weak suspension) then transferred to a rotary evaporator and continue to concentrate under reduced pressure. During the concentration add a few 1 liter portions of acetone until complete removal of the methylene chloride. The remaining acetone suspension (approximately 1 l volume) cooled overnight, and the crystals Marcfortine A collected and washed with several small portions of chilled acetone and dried under reduced pressure. Such crystals may contain in cachedtitle chloride as described) gives pure Marcfortine A.

Example 1A. Receipt and allocation of Marcfortine A and C

The primary fermentation seeds

Seed fermentati inoculant using agar plugs culture of Penicillium sp. IP 7780 (NRRL 18887), stored in liquid nitrogen. Three tubes are thawed and used as inoculum for 100 ml GS - 7 environment for planting. GS - 7 are from glucose and cotton seed flour (mark "Pharmamedia" by Traders Protein, Procter & Gamble Oilseed Products Co., Memphis, TN, U. S. A. adding each component at a concentration of 25 g/l of tap water. After composing pH GS - 7 was adjusted to 7.2 using NH4OH. Wednesday autoclave volume 100 ml 500 ml neprekonatelnych fermentation flasks for 30 minutes Sterile GS - 7 inoculant, as described above, and shaken at 250 rpm/min for 35-58 h, 23oC.

The method of carrying out fermentation (bulb vibrator)

Culture ripe seeds are used as inoculum for the production environment when 1% of the dose administration. The production environment consists of 45 g glucose, 25 g casein enzymatic digestion (brand Peptonized Milk Nutrient by Sheffield Products, Norwich, N. Y. U. S. A. 2.5 g yeast extract (brand Bacto Yeast Extract Code: 0127 by Difco Laboratories, Detroit, MI) for 1 litre of water. After establishing the pH of the produced medium is brought Dov 500 ml partitioned flasks for fermentation. Sterile environment production inoculant, as described above, and shaken 7-14 days at 250 rpm at 21oC.

The method of carrying out fermentation (collections Labraferm tanks)

Culture ripe seeds are used as inoculum for a sterile environment production at 0.5% of the dose administration. Environment production described above. After adjusting the pH of the medium to 7.0 using KOH 10 liters of this medium autoclaved for 90 min 12 l collections Labroferm tanks (New Brunswier Scientific Co., Inc.). Collections were inoculable at 0.5% of the dose of injection and stirred at 500 rpm at 20oC for 5-9 days. The flow rate of air is maintained between 10-15 l/min

The selection of Marcfortine A and C

West fermentative broth (35 l) which macerate at low speed in a large industrial mixer Waring'a and then mixed with an equal volume of methylene chloride. The mixture is left overnight under cooling and then centrifuged for breaking emulsions. The formed layer of transparent methylene chloride is drained and evaporated under reduced pressure. A concentrated solution of the residue (or 37.4 g) in methylene chloride is applied on a column Packed with a slurry of silica gel (1 kg), in methylene chloride. Column elute increasing concentrations acetonitrile from acetone to obtain Marcfortine A and Marcfortine C.

Example 1B. Receipt and allocation of Marcfortine A and C.

The method of fermentation seeds

Seed fermentati inoculant using agar plugs culture of Penicillium sp. IP 7780 (NRRL 18887), stored in liquid nitrogen. Three tubes are thawed and used as inoculum for 100 ml GS - 7 seed environment. GS - 7 amount of glucose flour from cotton seed (mark "Pharmamedia" by Traders Protein, Procter & Gamble Oilseed Products Co., Memphis, TN, U. S. A., by adding each component at a concentration of 25 g/l of tap water. After composing pH GS - 7 was adjusted to 7.2 using NH4OH. Wednesday autoclave in volumes of 100 ml in neprekonatelnych fermentation flasks for 30 minutes Sterile GS - 7 inoculant, as described above, and shaken at 250 rpm/min for 35 - 58 h at 23oC.

The method of carrying out fermentation (shake flask)

Culture ripe seeds are used as inoculum for the production environment when 1% of the dose administration. The production environment consists of glucose 20 g of glycerol 15 ml of flour cottonseed (mark "Pharmamedia" by Traders Protein, Procter Gamble Oilseed Products (o., Memphis, TN, U. S. A) 20 g soy flour 10 g, K2HPO43 g per 1 l of tap water. After establishing the pH of the medium producing adjusted to 6.8 using gidroponnyh flasks. Sterile environment production inoculant, as described above, and shaken for 7 - 14 days at 250 rpm at 21oC.

The method of carrying out fermentation (Labraferm collections)

Culture ripe seeds are used as inoculum for a sterile environment production at 0.5% of the dose administration. Environment production described above. After adjusting pH to 7.0 using KOH, 10 liters of this medium autoclave for 90 min 12 l collections (Labroferm tanks (New Brunswick Scientific Co., Inc). Collections inoculant at 0.5% of the dose of injection and stirred at 500 rpm at 20oC for 5 to 9 days. The flow rate of air support between 10 - 15 l/min

The selection of Marcfortine A and C

All fermentative broth (35 l) marcellous at low speed in a large industrial mixer Waring'a and then mixed with an equal volume of methylene chloride. The mixture is left overnight under cooling and then centrifuged for breaking emulsions. The formed layer of transparent methylene chloride is drained and evaporated under reduced pressure. A concentrated solution of the residue (or 37.4 g) in methylene chloride is applied on a column Packed with silikagelevye 91 kg) suspension in methylene chloride. Column elute increasing concentrations of acetone in IU the form from acetone to obtain Marcfortine A and Marcfortine C.

Example 2. 18-timecharter A (Compound 2)

A solution of 30 g marcfortine A and 22 mg of 2,4-bis(methoxyphenyl)-1,3-dithia-2,; -diphosphate-2,4-disulfide (reagent Lawesson's) in 5 ml of toluene is heated to boiling under reflux in nitrogen atmosphere for 18 hours the Mixture is cooled and the solvent is distilled off under reduced pressure. The residue is subjected to preparative chromatography (TCX) on plates of silica gel using 10% acetone in methylene chloride as eluent, to obtain 18-timecharter A. T. pl. 128 - 260oC.

FAB-MS 494 (M++ H)

In accordance with the General method of example 2, but using the appropriate marcfortine source material prepared following other compounds 18-timecharter:

1-methyl-18-timecharter A

1-benzyl-18-toumarkin A

1-ethyl-18-timecharter B

1-benzyl-18-timecharter B

18a-ethyl-18-timecharter B

18a-benzyl-18-timecharter B

18a-methoxyethoxymethyl-18-timecharter B

18a-allyl-18-timecharter B

18a-propargyl-18-timecharter B

1,18 a-bis-ethyl-18-timecharter B

1,18 a-bis-benzyl-18-timecharter B

18a-ethyl-24-methoxy-18-timecharter B

18-timecharter B (Compound # 2A)

FAB-MS 480 (M++ H)

18a-ethyl-24,2 marcfortine C

18a-benzyl-18-timecharter C

18a-methoxyethyl-18-timecharter C

18a-allyl-18-timecharter C

18a-propargyl-18-timecharter C

1,18 a-bis-propyl-18-timecharter C

1,18 a-bis-benzyl-18-timecharter C

18-timecharter C

1-Palmitoyl-18-timecharter A

1-decanoyl-18-timecharter A

1-decanoyl-18-timecharter B

Example 3. 24, 25 dihydro-18-timecharter A

A mixture consisting of 15 mg of 5% palladium on carbon and 18-timecharter A (30 mg, 0.6 mmol) in 1 ml of methanol, vigorously stirred in an atmosphere of hydrogen for 45 minutes, the Reaction mixture was filtered through Celiteand the filtrate evaporated under reduced pressure. Preparative thin layer chromatography of the residue on 0.5 mm plate of silica gel, eluruumi 3% methanol in methylene chloride, gives 24, 25-dihydro 18-timecharter A.

Example 4. 24,25-dibromo-24,25-dihydro 18-timecharter A

A solution of bromine in chloroform (0.4 ml 12M solution 0,048 mmol) is added dropwise to a chilled (ice bath) solution of 18-timecharter A (20 mg, 0.04 mmol) in 2 ml of chloroform. The resulting yellow solution was stirred at room temperature for 15 min and then evaporated in a stream of nitrogen. Preparative thin layer chromatography omarsdottir A.

Example 5. 24-methoxy-24,25-dihydro 18-tomarketing

A solution of bromine in chloroform (0.6 ml, 0.12 M solution 0,072 mmol) is added dropwise to a chilled (ice bath) solution of 18-tomarketing In (28 mg, 0.06 mmol) in 2 ml of chloroform. The resulting yellow solution was stirred at room temperature for 10 min, then at 0oC for 20 min, then evaporated in a stream of nitrogen. The yellow solid residue was dissolved in 2 ml of methanol, then add 1,8-diazabicyclo [5,4,0]undec-7-ene (DWI, of 0.015 ml, 0.10 mmol). The solution is stirred at room temperature for 90 min, then evaporated under reduced pressure. Preparative thin layer chromatography of the residue on 1.0 mm plate of silica gel, eluruumi 2% methanol in methylene chloride, gives 24-methoxy-25-bromo-24,25-dihydro-18-timecharter Century thus Obtained 24-methoxy-25-bromo-24,25-dihydro-18-timecharter dissolved In 2 ml dry toluene, then add tributyltin hydride (1.2 ml, 0.45 mmol). The solution was stirred at 100oC 16 h, then evaporated under reduced pressure. Preparative thin layer chromatography of the residue in 2.0 ml plate of silica gel, eluruumi 2% methanol in methylene chloride, gives 24-methoxy-24,25-dihydro-18-teamaction Century

Example 6. 24-ProPak the 24,25 - dihydro-18-tomarketing In (example 5), and the use of modified methods to 20 g 18-timecharter And gives an oily residue. Preparative conclusiona chromatography of the crude product on a 0.5 mm silica gel plate, aliremove 2% methanol in methylene chloride, gives 24-propoxy-24,25-dihydro-18-tomarketing A.

Example 7. 1-(dimethylcarbamoyl)-18-tomarketing AND

The potassium hydride (50 mg 25% oil dispersion) is added to a solution of 18-timecharter And (15 ml 0,030 mmol) in 1 ml dry tetrahydrofuran. The solution was stirred at room temperature for 2 h, then add dimethylcarbamoyl (0,028 ml, 0.30 mmol). The mixture is stirred at room temperature for 18 h, then partitioned between 5% aqueous sodium bicarbonate (1 ml) and methylene chloride (1 ml). The layers separated, and the aqueous layer was extracted with methylene chloride (2 ml). The combined extracts dried with magnesium sulfate, filtered and evaporated under reduced pressure. Preparative thin layer chromatography of the residue on a plate of silica gel, aliremove 25% acetone in hexane, to give 1-(dimethylcarbamoyl)-18-timecharter A.

According to example 7, but using the appropriate marcfortine starting material is prepared following other compounds 18 is propylboronic-18-timecharter AND

1-dimethylsulphamoyl-18-timecharter AND

1-(1-piperidinyl)thiocarbonyl-18-timecharter AND

1-succinoyl-18-teamaction AND

1-(2,4-dinitrobenzenesulfonyl)-18-timecharter AND

1-(4-morpholinomethyl-18-timecharter AND

1-(p-toluensulfonyl)-18-timecharter AND

1-acetyl-18-timecharter AND

1-methoxycarbonyl-18-timecharter AND

1-(p-toluensulfonyl)-18-timecharter

1-(2,4-dinitrobenzene sulfonyl)-18a-ethyl-24-methoxy-24,25-dihydro

1-(p-Brabanthal sulfonyl)-18-timecharter

1-propionyl 18-timecharter

1-(4-carbethoxy-1,3-thiazolidin-yl)carbonyl-18-timecharter A.

Example 8. 1-benzyl-18-thio-marchmain AND

The potassium hydride (75 ml 25% oil dispersion) is added to a solution of 18-tomarketing And (15 mg, 0,030 mmole) in 1 ml dry tetrahydrofuran. The solution is stirred at room temperature for 2 h, then add benzyl bromide (0,025 ml, 0.21 mmol). The mixture is stirred at room temperature for 3 h, then partitioned between water (1 ml) and methylene chloride (1 ml). The layers are separated and the aqueous layer was extracted with methylene chloride (2 ml). The combined extracts dried with magnesium sulfate, filtered and evaporated under reduced pressure. Preparative thin-layer chromatography is Fiorini A. According to example 8, but using the appropriate marcfortine source drug cook below other compounds 18-tomarketing:

1, 18a-bis-ethylene-18-timecharter;

1-allyl-18-timecharter A;

1-methoxymethyl-18-timecharter A;

1-propargyl-18-timecharter A;

1-benzyl-18a-Teal-18-timecharter;

Example 9. Halmonchus contortus /Trichostrongylus colubriformis / Tird Assay:

This in vivo (in vivo) testing use of gerbils infected with two important target parasites of ruminants, H. contortus (H. C) and T. colubriformis (T. C) (can be used in sensitive versus resistant worms or worms). Initially, the activity determined only against H. contortus, as described in G. A. Conder et al, T. Parasitol, 76, 168-170(1990), whereas in the later stages of the learning activity against both species of parasites, using the techniques described in G. A. Conderetal., T. Parasitol. 77, 621-623 (1991). Activity the following table. 2.

Example 10. According to example 2, but using the appropriate paraphernalia/typeahead source drug cook below other compounds 18-thiomargarita:

18-typeahead

1-methyl-18-typeahead

1-ethyl-18-typeahead

Example 10A. According to example 7, but using the appropriate typeahead source drug cook below other compounds 18-thiomargarita:

1-acetoxymethyl-18-tomarketing

1-diethoxyphosphoryl-18-typeahead

1-cyclopropanecarbonyl-18-typeahead

1-dimethylsulphamoyl-18-typeahead

1-(1-piperidinyl)thiocarbonyl-18-typeahead

1-succinyl-18-typeahead

1-(2,4-dinitrobenzenesulfonyl)-18-typeahead

1-(4-morpholinomethyl)-18-typeahead

1-(p-toluensulfonyl)-18-typologically

1-acetyl-18-typeahead

1-methoxycarbonyl-18-typeahead

Example 11. According to example 8, but using the appropriate paraphernalia starting material is prepared following other compounds

18-thiomargarita:

1,18-bis-ethyl-18-typeahead

1-allyl-18-typeahead;

1-methoxymethyl-18-typeahead;

1-propargyl-18-typeahead;

1-benzyl-18a-ethyl-18-typeahead.

Example 12. 1-pyrrolidinyl-marcfortine And connect. 12.

The potassium hydride (50 mg 25% oil dispersion) is added to a solution of marcfortine who make pyrrolidinylcarbonyl (40 mg, 0.30 mmol). The mixture is stirred at room temperature for 18 h, then partitioned between 5% aqueous sodium bicarbonate (1 ml) and methylene chloride (1 ml). The layers are separated and the aqueous layer was extracted with methylene chloride (2 ml). The combined extracts dried with magnesium sulfate, filtered and evaporated under reduced pressure. Preparative thin layer chromatography on a plate of silica gel, aliremove 25% acetone in hexane, gives 1-pyrrolidinyl carbonyl-marcfortine AND

Here and further in the text:

1) in the spectra of1H-NMR (TMR)

s - singlet - brs broadened singlet

m - multiplet

d - doublet

t - triplet

dd = double doublet

2) the Item. analysis:

calc - calculated

obsd - found

H1NMR(CDCl3) of 0.83(s, 3H), 1,21(s, 3H), 1,45(s, 3H), 1,3-2,4(m, 17H), of 2.38(d, 1H), 2,6-2,8(m, 2H), 2,84(d, 1H), 3.00 and(s, 3H), 3,4-3,6(m, 4H), and 3.72(d, 1H), around 4.85(d, 1H), 6,41(d, 1H), 6,84(s, 2H)< / BR>
In accordance with the General method of example 12, but using the appropriate makhorin the source of the drug and the carbonyl chloride prepared following other compounds makhorin:

1 - diethoxyphosphoryl - Makhotin AND

1 - cyclopropanecarbonyl - marcfortine And, Conn. #12A.

1H NMR (CDCl3) of 0.82(s, 3H), 1.05(s, 3H), of 1.05(s, 3H), of 1.44(2s, 6H), of 0.9 to 1.7(m, 8H), 1,7-3,0(m, 11H), 3,14(s, 3H), 3,80(d

1-phenoxycarbonyl-marcfortine And Conn. #12B.

1H NMR (CDCl3) of 0.83(s, 3H), of 1.09(s, 3H), 1.3 to 1.5(m, 1H), 1,42(s, 3H), of 1.46(s, 3H), 1.5 to 3.0(m, 13H), 3,14(s, 3H), 3,70(d, 1H), a 4.83(d, 1H), 6,27(d, 1H), 6.90 to(3, 2H), 7,2-7,5(m, 5H).

1-(2,4-dichlorophenoxyacetic)-18a-allyl-marcfortine B

1-(4-oxopentanoic)-18-methyl-marcfortine C

1-Palmitoyl-marcfortine And, Conn. #12C

FABS-MS 716(M++ H)

1-(4-nitrophenoxyacetic)-marcfortine And, Conn. # 12D1H NMR (CDCl3) of 0.90(s, 3H), of 1.09(s, 3H), 1.3 to 1.5(m, 1H), 1,44(s, 3H), of 1.47(s, 3H), 1,5-2,9(m, 13H), 3,14(s, 3H), 3,70(d, 1H), a 4.86(d, 1H), 6,23(d, 1H), 6,93(ABg, 2H), of 7.48(d, 2H), 8,32(d, 2H).

1-(1-piperidinylcarbonyl)marcfortine And, Conn. #12F

FAB-MS 589(M++ H); H RNS m/z (M++ H, C34H44N4O5+ H1) calc 589.3390, obsd 589.3398

1-[[4-(etoxycarbonyl)piperazine-1-yl] carbonyl] - marcfortine And, Conn. # 12G

FAB-MS 662(M++ H); 684 (M++ Na); HRMS m/z (M++ H, C36H47N5O7+ H1), calc 662,3553, obsd 662.3569

1-[[4-(benzyl)piperazine-1-yl]carbonyl] - marcfortine And, Conn. # 12H

1H NMR (300mHz, CDCl3):0,80(s, 3H), 1.26 in(s, 3H), of 1.41(s, 3H), 1,45(s, 3H), 1,2-2,9(m, H), 2,98(s, 3H), of 3.4 to 3.8(m, 8H), 6,41(d, 1H), 6,84(s, 2H), 7,2-7,5(m, 5H), FAB-MS 680 (M++ H); 702(M++ Na)

1-[[4-(1,3-benzodioxol-5-ylmethyl)piperazine-1-yl] carbonyl] - marcfortine And, Conn. # 12I

1H NMR (300m Hz, CDCl3): 0,80(s, 3H), 1.00 and-2,60(m, 15H), metal)piperazin-1-carbonyl]-marcfortine And, Conn. # 12J

1H NMR (300mHz, CDCl3):0.82 (s, 3H),1.26 (s, 3H), 1.40 - 2.72 (m, 20H), 1.41 (s, 3H), 1.44 (s, 3H), 2.82 (d, 1H) 2.99 (s, 3H), 3.40-3.81 (m, 5H), 4.85 (d, 1H), 6.41 (d, 1H), 6.84 (s, 2H). FAB-MS 604 (M++H)$ HRMS m/z (M++H, C4H45N5O5+H1), Calc 604.30499, obsd 604.3506

1-(1,3-dihydro-1,3-dioxo-2H-isoindole-2-yl)-marcfortine And, Conn. # 12K FABS-MS 665 (VH)

1-[[4-pyridin-2-yl)piperazine-1-yl] carbonyl] -marcfortine And, Conn. 12 # L1H NMR (300m Hz, CDCl3) : 0,84 (s, 3H), 1.10-2.30 (m, 12H), 1.41 (s, 3H), 1.44 (s, 3H), 2.40 (d, 2H), 2.67 (t, 2H), 2.85 (d, 1H), 3.00 (s, 3H), 3.45-3.85 (m, 9H), 4.85 (d, 1H), 6.40 (d, 1H). 6.60-6.70 (m, 2H), 6.85 (s, 2H), 7.52 (t, 1H), 8,15-8.22 (m. 1H), FAB-MS 667 (M++H)

1-[[4-(phenyl)piperazine-1-yl] carbonyl] -marcfortine And, Conn. # 12M1H NMR (300 m Hz, CDCl3): 0,84(s, 3H), 1.20-2.50 (m, 11H), 1.22 (s, 3H), 1.42 (s, 3H), 1.45 (s, 3H), 2.67 (t, 1H), 2.85 (d, 1H), 3.00 (s, 3H), 3.10-3.91 (m, 9H), 4.85 (d, 1H), 6.40 (d, 1H), 6.85 (s, 2H), 6.90-7.00 (m, 3H), 7.25-7.38 (m, 2H)

1-[[4-(chlorocarbonyl)piperazine-1-yl]carbonyl]-marcfortine A. Conn. # 12N

1H NMR (300m Hz, CDCl3): 0.74 (s, 3H), 1.18 (s, 3H), 1.30-2.65 (m. 20H), 2.74 (d, 1H), 2.81 (s, 3H), 3.48-3.90 (m, 8H), 4.30 (d, 1H), 6.32 (d, 1H), 6.80 (s, 2H), FAB-MS 652 (M++H)

10,10" - (1,4-decarbonylation)bis(6', 7',8',9',10',10' hexahydro-1', 1', 4,4,12 - pentamethyl)-[2'S - [2' - alpha, 3'a, alpha, 9'a alpha, 10/2"' R*, 3"'a S*, 10"'a R*), 10'a. beta]] - S Piro [4H, 8H - [1,4] dioxano [2,3-g]indole - 8,2' (3 n) - [1H, 4H - 3a, 9a] (aminomethane) of cyclopent [b] hearten And, Conn. #12P1H NMR (300m Hz, CDCl3); 0.76 (s, 3H) 1.10 - 2.30 (m, 13H), 1.14 (s, 3H), 1.35 (s, 3H), 1.38 (s, 3H), 2.33 (d, 1H), 2.52-3.00 (m, 8H), 3.00 - 3.20 (m, 1H), 3.50 - 3.75 (m, 2H), 4.05 (d, 1H), 4.33 (d, 1H), 4.79 (dd, 1H), 6.35 (dd, 1H), 6.78 (s, 2H), 7.10 - 7.30 (m, 5H). FAB-MS 665 (M++ H)

1-[[4-(dimethyl)piperidine-1-yl] carbonyl] - marcfortine A, Conn. 12Q1H NMR(300m Hz, CDCl3); 0.75 (s, 3H), 0.90 - 1.95 (m, 13H), 0.92 (s, 3H), 0.96 (s, 3H), 1.14 (s, 3H), 1.34 (s, 3H), 1.37 (s,3H), 2.08 (d, 1H), 2.20 (brs, 1H), 2.33 (d, 1H), 2.60 (t, 2H), 2.77 (d, 1H), 2.92 (s, 3H), 3.45 - 3.75 (m, 4H), 4.77 (d, 1H), 6.33 (d, 1H), 6.77 (s, 2H). FAB-MS 617( M++ H)

1-[[4-(5-chloropyridin-3-yl)piperazine-1-yl] carbonyl] - marcfortine And, Conn. 12 R1H NMR (300m Hz, CDCl3): 0.83 (s, 3H), 1.20-2.30 (m, 11H), 1.22 (s, 3H), 1.42 (s, 3H), 1.44 (s, 3H), 2.45 (d, 1H), 2.65 (t, 2H), 2.83 (s, 1H), 2.99 (s, 3H), 3.60-3.85 (m, 8H), 4.86 (d, 1H), 6.40 (d, 1H), 6.86 (s, 2H), 6.92 (d, 1H), 7.29 (d, 1H). FAB-MS 702 (M++ H).

1-[[4-course)piperazine-1-yl] carbonyl] - marcfortine A, Conn.# 12S1HNMR (300m Hz, CDCl3); 0.84(s, 3H), 1.20 - 2.30 (m, 9H), 1.26(s, 3H), 1.42 (s, 3H), 1.45 (s, 3H), 2.40 (d, 1H), 2.60 - 2.75 (m, 2H), 2.85 (d, 1H), 3.00 (s, 3H), 3.15 - 3.30 (m, 4H), 3.49(d, 1H), 3.52 - 3.84 (m, 5H), 4.85 (d, 1H), 6.39 (d, 1H), 3.15 - 3.30 is (m, 4H), 3.49 (d, 1H), 3.52 - 3.84 (m, 5H), 4.85 (d, 1H), 6.39 (d, 1H), 6.80 - 6.90 (m, 4H), 7.23 (d, 2H).

10'10"-(1,4-dioxo-2-butene)bis(6', 7', 8', 9', 10'a - hexahydro- 1',1', 4,4,12'-pentamethyl)- [2'S - 2 alpha, 3'a alpha, 3'a alpha, 9'a, alpha, 10(2"'R*, 3"' a S*9"'a S*, 10"'a R*), 10"a beta]]-S Piro[4H, 8H-[1,4] dioxano[2,3-g] indole-8,2'(3'H)-[1H, 4H-3a, 9a](imename the piperazine-1-yl] carbonyl] -marcfortine A, Conn.# 12U

1H NMR (300m Hz, CDCL3: 0.75 (s,3H), 1.1-1.85 (m,8H), 1.15 (s, 3H), 1.34 (s,3H), 1.34 (s,3H), 1.92 (d,1H), 2.00-2.40(m,2H), 2.34 (d,1H), 2.55 (t,24), 2.7 b (d,1H), 2.91 (s,3H), 3.42-3.75 (m, 8H), 4.65-4.77 (m,2H), 4.79 (d,1H), 6.33 (d,1H), 6.79 (s,2H), FAB-MS 764 (M+H); HRMS m/z (M++ H, C6H44Cl3N5O7+H1), alc 764.2384, obsd 764.2366

1-[[4-(3-trifluoromethyl-thiadiazole-yl)piperazine-1-yl] carbonyl]-marcfortine A, Conn.# 12V

NMR (300m Hz, CDCl3): 0.83 (s,3H), 1.26 (s,34), 1.30-2.30 (m,11H), 1.42 (s, 3H), 1.45 (S,3H), 2.42 (d,1H), 2.55-2.72 (m,2H), 2.84 (d,1H), 2.99 (S, 3H), 3.60-3.90 (m,8H), 4.87 (d,1H), 6.30 (d,1H), 6.88 (s,2H), FAB-MS742 (M++ H); HRMS m/z (M++ H, C36H42F3N7S1+H1), calc 742.2998, obcd 742.3038

Example 13. 1-acetoxymethyl-marcfortine A, Conn.#13

Sodium hydride (467 mg of 60% oil dispersion) is added to a solution of marcfortine A(850 mg, 1.78 mmol) in 1 ml of tetrahydrofuran. The solution was stirred at room temperature for 4 h and then add methyl bromoacetate (280 mg, 1.82 mmol). The mixture is stirred at room temperature for 8 h, then partitioned between 5% sodium bicarbonate (1 ml) and methylene chloride (1 ml). The layers are separated and the aqueous layer was extracted with methylene chloride (2 ml). The combined extracts dried with magnesium sulfate, filtered and evaporated under reduced pressure. Chromatography on a plate siliconera, e is to maintain oil:

1H NMR (CDCl3) 0.75 (3H, s), 1.08 (3H, s), 1.43 (3H, s), 1.45 (3H, s), 2.05 (3H, s), 3.12 (3H, s), 4.86 (1H, d, J = 8), to 5.85 (1H, d, J = 8), 5.96 (1H, d, J = 8), 6.26 (1H, d, J = 8), 6.77 (1H, d, J = 7), 6.86 (1H, d, J = 7).

According to example 13, but using the appropriate marcfortine starting material is prepared following other compounds marcfortine:

1-t-butyryloxy-marcfortine A

1-benzoyloxymethyl-marcfortine A

1-t-butyryloxy-marcfortine A

1-acetoxymethyl-18a-benzyl marcfortine B

1-3,4-dichlorobenzoate-marcfortine C

Example 14. 1-(2,4-dinitrobenzenesulfonyl)-marcfortine A, Conn. 14

Marcfortine A (75 mg, about,15 mmol) are added to a suspension of potassium hydride (180 mg, 1.50 mmol, 35 wt.% in raises. oil) in THF (6 ml) at 5oC, followed by addition of 2,4-dinitrobenzenesulfonyl chloride (73 mg, 0.30 mmol). The reaction mixture is then stirred for 3 h at 5oC. After dilution with water and extraction with chloroform, the crude product chromatografic 1 mm plate of silica gel (TLC), elwira 30% acetone in methylene chloride, getting 1-(2,4-dinitrobenzenesulfonyl)-marcfortine A (90 mg, 89,1%) as a yellow solid product. So pl. 155 - 160oC (decomp.).

1H NMR (CDCl3); 0.86 - 0.97 (m, 3H), 1.13 - 1.45 (m, 15H), 1.56 - 1.78 (m, 5H), 1.86 (m, 1H), 2.00 (m, 1H), 2.15 (d, 1H), 2. 9.16 (d, 1H).

According to example 14, but using the appropriate marcfortine starting material is prepared following other compounds marcfortine:

1-(4-morpholinomethyl)-marcfortine A, Conn. # 14A

FAB-MS; m/e 595 (m++ H)

1-(trichloronitromethane)-marcfortine A, Conn. # 14B

FAB-MS; m/e 626, 628, 630 (M++ H)

1-(methoxycarbonylmethyl)-marcfortine A, Conn. # 14C)

FAB-MS; m/e 568 (M++ H)

1-(benzazolyl)-marcfortine A, Conn. # 14D

FAB-MS; m/e 586 (M++ H)

1-trichloronitromethane-18a-ethyl-marcfortine B

1-ethoxycarbonylmethyl-18a-methyl-marcfortine C

Example 15. 1-(4-morpholinylcarbonyl)-marcfortine A N-oxide, Conn. # 15

1-(4-Morpholinoethyl)-marcfortine A (Conn. # 12E, 15 mg) is treated with 3-chloronitrobenzene acid (15 mg) in methylene chloride (2 ml) at 0oC for 10 minutes the Mixture was partitioned between 5% sodium bicarbonate (2 ml) and methylene chloride (2 ml). The layers are separated and the aqueous layer was extracted with methylene chloride (2 ml). The combined extracts dried with magnesium sulfate, filtered and evaporated under reduced pressure. The remainder chromatographic 0.5 mm plate of silica gel preparative thin-layer chromatography, elwira 10% methanol in METI is="ptx2">

Example 16 1-(2-tetrahydrofuranyl)marcfortine A, Conn. # 16

Carbonyldiimidazole (50 mg) are added to a solution marcfortine A (60 mg) in 6 ml of tetrahydrofuran. The mixture is heated to boiling under reflux and add potassium hydride (120 mg). The resulting mixture is stirred, continuing to boil under reflux for 1 hour the Precipitate is filtered off and the filtrate concentrated. The remainder chromatographic on a plate of silica gel, elwira 25% acetone in methylene chloride, and get named in the title compound (30 mg) as a solid product. HRM S (FAB); m/e (M++ H), C32H41N3O5+ H) calc. 548, 3124, obsd. 548, 3086.

Example 17. 1-(4-morpholinylcarbonyl)-marcfortine A, Conn. # 12E

Marcfortine A (1 g, 2.1 mmol) in THF (25 ml, purified by distillation over metallic sodium from benzophenone) and potassium hydride (35 wt.% 0.5 g, 4.4 mmol) is stirred for 0.5 h at room temperature in a nitrogen atmosphere. Then using a syringe add morpholinylcarbonyl chloride (1 ml, 8.4 mmol). After 3 h stirring at room temperature, the turbid reaction mixture is cooled in an ice bath and quickly stop the reaction by adding dropwise a cooled saturated solution of sodium carbonate (5 ml). The mixture is then diluted with water (75 ml) and tryout obtain the crude product, containing oil. Chromatography on silica gel (2% to 5% MeOH/CH2Cl2) gives 1-morpholinylcarbonyl-marcfortine A. (Chromatography on silica gel on chromatotron (4% MeOH/CH2Cl2) is the best method of cleaning for 1-morpholinylcarbonyl-marcfortine)

1H NMR (300 MHz, CDCl3); 0.81 (s, 3H), 1.21 (s, 3H), 1.19 - 1.94 (m, 8H), 1.41 (s, 3H), 1.45 (s, 3H), 1.98 (d, 1H), 2.08 - 2.28 (m, 2H), 2.40 (d, 1H), 2.62 (t, 2H), 2.82 (d, 1H), 2.99 (s, 3H), 3.50 - 3.86 (m, 8H), 4.86 (d, 1H), 6.41 (d, 1H), 6.85 (s, 2H), FAB-MS 591 (M++ H); HRMS M/Z (M++ H, C33H42N4O6+ H1), calc. 591.3182, obsd 591. 3200.

Example 18. According to example 12, but using the appropriate paraphernalia starting material is prepared following other compounds paraphernalia:

1-diethoxyphosphoryl-paraphernalia

1-cyclopropanecarbonyl-preocupied

1-(1-piperidinyl)thiocarbonyl-paraphernalia

1-succinoyl-paraphernalia

1-(4-carbethoxy-1,3-thiazolidin-3-yl)carbonyl-paraphernalia

1-phenoxycarbonyl-paraphernalia

1-pyrrolidinyl-paraphernalia

1-piperidinylcarbonyl-paraphernalia

1-(4-morpholinylcarbonyl)-paraphernalia

Example 19. According to example 13, but using the appropriate paraphernalia source material purchasing to proximity-paraphernalia

1-acetoxymethyl-18a-paraphernalia

1-3,4-dichlorobenzoate-paraphernalia

Example 20. According to example 14, but using the appropriate paraphernalia source material, prepared following other compounds paraphernalia:

1-(2,4-dinitrobenzenesulfonyl)-paraphernalia

1-(4-morpholinomethyl)-paraphernalia

1-trichloronitromethane-18a-ethyl-paraphernalia

1-ethoxycarbonylmethyl-18a-methyl-paraphernalia

1-Palmitoyl-paraphernalia

Methodology A

A. Seed fermentat Cunninghamtlla cchinulata subsp elegans (-) NRRL 1368, Cunningha-mella blakesleeana (+) Atcc SbSSa, Cunnirghamella echinulata subsp. sllgans (-) NRRL 1393, Cunnirghamella echinulata NRRL 3655.

Frozen agar tube NRRL 1368 or 1393, ACTS 8688a or preferably NRRL 3655 (stored in liquid nitrogen LN2") is aseptically transferred into a 100 ml volume of GS-7 (sterile) environment, contained in a 500 ml wide-mouthed flask for shaking.

GS-7 are from (Cerelose) and Pharmamelig (Pharmacracy), each added at a concentration of 25 g/l of tap water. the pH of the medium was adjusted to 7.2 using ammonium hydroxide. The medium is sterilized by autoclaving for 30 minutes Inoculated GS-7 shake at 150-300 (preferably 250 rpm) at 21-30oC, preferably coleosperma at approximately 1 to 5% of the dose (preferably about 5% of the dose).

B. Method biotransformation

100 ml volumes of GS-7, contained in a 500 ml wide-mouthed flasks, for fermentata inoculant 48-72 h seed fermentation at approximately 1 - 5% of the dose (preferably about 5% of the dose). Inoculated CS-7 shaken at approximately 250 rpm at 28oC for 24-48 hours During this time to fermentate add Marcfortine A, dissolved in dimethylformamide (DMF), in the interval between 10-12 mg/flask, preferably at 10 mg in 0.4 ml of DMF in a 100 ml flask/fermentat. Fermentation in the presence of Marcfortine A continue, as indicated previously, within 24-72 hours

Contents 100 flasks, each containing 100 ml of fermentata, as described in the method for biotransformation, which was shaken at 28oC for 24-72 h followed by the addition Marcfortine A (0,010 g per flask, the total amount of 1 g) in DMF (0.2 ml per flask, 20 ml), treated with CH2Cl2(100 ml) to each flask. The contents are combined and stirred wacihg mixer 5 minutes the Mixture centrifugum decanted and the resulting aqueous layer, the Organic layer collected, dried (sodium sulfate), filtered and concentrated. The components of the crude mixture divided by a series of column chromatography, using a ratio of the silica gel - solid extract aromatherapies fractions separated Marcfortine B. HRMS(FAB); m/e found: 4642563; calculated: C27H33N3)O4+ H, 4642549.

If desired marcfortine B can be recrystallized from a mixture of methylene chloride/acetone. For example, the solution marcfortine B in methylene chloride concentrate at a residual pressure on a rotary evaporator. The concentrate is diluted portions of acetone during the concentration, until a suspension marcfortine B in acetone.

1. Derivatives marcfortine/paraphernalia formula I

< / BR>
where m = 0 or 1;

n = 1;

W = S or O;

R14andand R14bis a hydrogen atom;

R24and R25form a double bond;

R18andis a hydrogen atom or a C1- C7- alkyl;

R1is a hydrogen atom, -C(O)R1group (where R1- cyclo3- C8-alkyl), phenoxycarbonyl, -C(O)-C2-C24-alkyl, nitrophenoxyacetic, 1,3-dioxo-2H-isoindolyl, dinitrobenzenesulfonyl, trichloromethylsulfuryl,1- C4-alkoxycarbonylmethyl, benzazolyl, tetrahydropyranyl, 1,4-dioxy-C2-C7alkenyl or R1group-C(O)NR4R5where R4and R5together with the nitrogen atom form nennig1- C4-alkyl, C1-C4-alkanoyl, C1-C4-alkoxycarbonyl, 1,3-benzodioxolyl-C1- C4-alkyl, pyridinyl, halogenopyrimidines, phenyl (optionally substituted by halogen atom), chlorocarbonyl, trifter-C1-C4-alkoxycarbonyl, trifter-C1-C4-alkyldiethanolamine, or R4and R5form piperidinyl, optionally substituted phenyl or mono - or disubstituted WITH1-C4-alkyl,

or their pharmaceutically acceptable salts, or hydrates.

2. Connection on p. 1, characterized in that the group R1represents - and - a saturated heterocyclic ring, as defined in paragraph 1.

3. Connection on p. 2, where a saturated heterocyclic ring selected from 4-research, 4-phenyl-1-piperazine, 4-(2-pyridinyl)-1-piperazine, 2,6-dimethyl-4-research, 1-pyrrolidino, 4-methyl-1-piperazine, 1-piperidine, 4-phenyl-1-piperidine, 4-phenylpiperidine, 2-methylpiperidine, 3-methylpiperidine, 4-methylpiperidine, N-methylpiperidine, 1-methylhomopiperazine, 1-acetylpiperidine.

4. Connection PP.1 to 3, with deworming and acaricide properties.

5. The method of suppressing insects or nematodes in the media, characterized in that as the active component connection is used on the PP.1 - 3.

6. The strain of the fungus Penicillium sp. NRRL 18887, producer marcfortine and/or marcfortine C.

7. Connection on p. 1, characterized in that it is a

18-timecharter A (compound 2),

18-timecharter (compound 2A),

1-pyrrolidinyl-marcfortine And (compound 12),

1-cyclopropanecarbonyl-marcfortine And (compound 12A),

1-phenoxycarbonyl-marcfortine And (compound 12B),

1-Palmitoyl-marcfortine And (compound 12C),

1-(4-nitrophenoxyacetic)-marcfortine And (compound 12D),

1-(1-piperidinylcarbonyl)-marcfortine And (compound 12F),

1-[[4-(etoxycarbonyl)piperazine 1-yl]carbonyl]-marcfortine And (compound 12G),

1-[[4-(benzyl)piperazine-1-yl] carbonyl]-marcfortine (connection N),

1-[[4-(1,3-benzodioxol-5-ylmethyl)piperazine-1-yl] carbonyl]-marcfortine And (compound 12I), 1-[[4-(methyl)piperazine-1-yl] carbonyl] -marcfortine And (compound 12J),

1-(1,3-dihydro-1,3-dioxo-2H-isoindole-2-yl)-marcfortine (connection 12K),

1-[[4-(pyridin-2-yl)piperazine-1-yl] carbonyl] -marcfortine (connection 12L),

1-[[4-(phenyl)piperazine-1-yl] carbonyl]-marcfortine (connection 12M),

1-[[4-chloro-carbonyl)piperazine-1',4,4,12'-pentamethyl)-[2'S-[2'. alpha, 3 a.alpha, 9 a.alpha,10] 2"' R*, 3"'as*9"'as, 10"'ar*)-10 a. beta]]-Spiro[4H,8H-(1,4-doxepin)- 2,3-g[indole-8,2'-(3 N)] 1H, 4H-3A,9a[(aminomethane of cyclopent [b] hemolysin] -9,11'-(10H)-dione (compound 12°), 1-[[4-(phenyl)piperidine-1-yl] carbonyl] -marcfortine (connection R),

1-[[4-(dimethyl)piperidine-1-yl]carbonyl]-marcfortine (connection 12Q),

1-[[4-(5-chloropyridin-3-yl)piperazine-1-yl] carbonyl] -marcfortine (connection 12R),

1-[[4-(4-course)piperazine - 1-yl] carbonyl]-marcfortine (connection 12S),

10,10"-(1,4-dioxo-2-butene)bis(6', 7',8',9',10',10A-hexahydro-1',1', 4,4,12'-pentamethyl)-[2'S-[2'.alpha, 3 a.alpha, 9 a.alpha, 10(2"'R*, 3"'as*9"' as*, 10"'ar*) 10 a.beta]]-Spiro[4H,8H-(1,4-doxepin)-2,3 - g[indole-8,2'(3 N)-1H, 4H-3A, 9a] (aminomethane)of cyclopent [b] hemolysin]-9,11'-(10H)-dione (compound 12T),

1-[[4-(Trichloroisocyanuric) piperazine-1-yl] carbonyl] -marcfortine And (compound 12U),

1-[[4-(3-trifluromethyl-thiadiazole-1-yl)piperazine-1-yl] carbonyl)-marcfortine a (12V connection).

 

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