Derived embryo death, pharmaceutical or veterinary composition and method of producing compounds

 

(57) Abstract:

In the present invention proposed derived embryo death of the formula I, with antiparasitical activity

in which the dotted line indicates an optional carbon-carbon bond, R1H or OH, R2-C1-C8alkyl, C3-C8-cycloalkyl, R3Is H or OH, R4-H or a group capable of either hydrolyzed in vivo, as well as the way they are received, intermediate compounds, pharmaceutical or veterinary composition comprising an effective amount of the compounds of formula I and an acceptable excipient or carrier. 3 s and 5 C.p. f-crystals, 1 table.

The present invention relates to new antiparasitics funds related milbemycin and avermectins, methods for their preparation and their preparations.

The avermectins are a group of antiparasitics funds a wide spectrum of action, previously called compounds C-076. Get them fermentation involving strain of the microorganism Streptomyces avermitilis in aerobic conditions in an aqueous nutrient medium containing inorganic salts and Assemblywoman sources of carbon and nitrogen. Isolation and chemical structure of the eight individual components, kotori contains eight different but closely related compounds, described as C-076 A1a, A1b, A2a, A2b, B1a, B1b, B2a and B2b. A number of compounds refers to the natural avermectins, whose 25 - Deputy is (S) - sec-bootrom, and the number "b" refers to those compounds, in which the 25-Deputy is isopropyl. The designations "A" and "B" refer to the avermectins have a 5 Deputy is a methoxy group or a hydroxy-group, respectively, and the numeral "1" refers to the avermectins, in which the double bond is present at position 22(23), the number "2" refers to the avermectins, which are not 22 (23)-double bond and contain hydrogen in the 22-position and a hydroxy-group at the 23 - position.

In our applications for European patents NN 0214731, 0284176, 0317148, 0308145, 0340832, 0335541 and 0350187 describes methods of producing compounds related to the avermectins, but with a 25-position a group other than isopropyl or (S) - sec-butyl found in the original avermectins specified in the description of the patent England N 1573955. Such compounds can be obtained by fermentation with the participation of certain strains of Streptomyces avermitilis in the presence of organic acids or their derivatives. The receipt of such avermectins described in Journal of Antibiotics (1991), 44, N 3, pp. 357-365.

Milbemycin form another MgO in the C-13 position. Examples of such compounds are described in patent England N 1390336 and European patent publications NN 170006, 254583, 334484 and 410615. In addition to these fermentation products a large number of publications describes compounds derived semi-synthetic of these fermentation products, many of which have suitable antiparasitical activity. Part of these methods is discussed in Maerolide Antibiotics, Omura, S., E. d., Academic press, New York (1984) and Davies, H. G., Green, R. H. B Natural product Reports (1986), 3, 87-121 and in Chem. Soc. Rev., 1991, 20, 271-339.

It has been shown that some compounds that can be obtained synthetically from known avermectins avermectins and derivatives, have unexpectedly useful biological properties.

In accordance with one aspect of the present invention proposed the compounds of formula 1

< / BR>
in which the dashed line in 22 (23)- position indicates a possible additional bond, and, or this relationship is present and R1missing, or this relationship is absent and R1represents H or OH, R2represents a C1-C8-alkyl or C3-C8-cycloalkyl,

R3represents H,

R4represents H or a group capable of either hydrolyzed in vivo

If the context is not explicitly stated, all the alkyl and alkeneamine substituents having 3 or more carbon atoms, may have a normal chain or branched structure. The term "aryl" includes phenyl, which may be substituted by at least one C1-C6-alkyl, hydroxy-group, C1-C6-alkoxygroup, halogen, the nitro-group or CF3. In the present invention the term "alkyl" refers to alkali with 1-8 carbon atoms, for example methyl, ethyl, propyl, isopropyl, butyl, pentyl, hexyl and the like alkali, with a normal chain or branched structure. The term "alkanoyl" means alkanoyl with 1-8 carbon atoms, for example formyl, acetyl, propionyl, butyryl, pentanoyl, hexanoyl and similar radicals.

The term "carbarnoyl" denotes the group-CONR7R8in which R7and R8identical or different, represent H, alkyl, aryl, heteroaryl or form a 4-8-membered nucleus containing one or more atoms O, N or s

Group hydrolyzable in vivo with the formation of the corresponding compounds in which the group is substituted by H, is generally well known in pharmacy, a number of such groups suitable for use in compounds of the present invention. PR is the remains of dicarboxylic acids and amino acids. Specific such groups are indicated in the examples below. Preferred are those compounds in which R2- cyclohexyl, R3represents H and a possible link in 22 (23)-position is absent and R1represents H.

In particular, the preferred oximes of monosaccharides, in which R4represents H.

Individual compounds of the invention are described in the examples below.

The most preferred compound is a monosaccharide-5-oximino-22,23-dihydro - 25-cyclohexylaniline B1.

In accordance with another aspect of the invention, a method for obtaining such compounds, which includes stages: (i) oxidation of compounds of formula II

in which the dotted line R1, R2, R3and R6has the above significance, and R5has the above values or R5is a - - alejandroisaacura;

for the formation of compounds of formula III

< / BR>
and (ii) the reaction of compounds of formula III with the compound of the formula

R4- O - NH2,

where

R4has the above values, and

R5represents alejandroisaacura,

and gidrospetstroy a H, the group can either hydrolyzed in vivo to form compounds in which R4represents H. If necessary (iv) (vii) hydrogenation of compounds to restore the double bond in position 22 (23) in a single bond.

Obtaining compounds of the invention is discussed and illustrated below.

Compounds of the present invention can be obtained from compounds of formula (iv), which can be obtained as described in the above patent publications.

< / BR>
Connection PA. The double bond is present, R1is missing.

Connection b. The double bond is absent, R1= H.

The Connection Of The Substation. The double bond is absent, R1= OH.

Semi-synthetic modifications required to obtain the compounds of formula I, may require a sequential reaction, the exact procedure for such transformations may change. Connection with the 23-hydroxy-group (or its protected derivative), can be transformed into the corresponding 22, 23-Dihydrocodeine or the corresponding compound with a double bond in 22(23)- position, using the methods described in U.S. patent N 4328335. The latter compound can also hydrogenation to turn 22, 23-d is P> Obtain the compounds of the invention can be achieved primarily by conversion of the above disaccharides PA, b and c in their respective monosaccharides by hydrolysis. Another way to obtain monosaccharides consists of direct fermentation of the corresponding aglycone, as described in the application for the European patent N 463677.

According to another method, compounds of the invention can be obtained by carrying out the above synthetic transformations on the disaccharides PA, b or C and subsequent hydrolysis at the end of the target monosaccharides.

If desired, the hydroxy-group can be allievate to obtain esters with the use of such reagents as anhydrides or acid chlorides and amines acids in accordance with commonly known in this field techniques. Hydroxy-group can be converted into the carbonyl group by oxidation dioxide magnesium or perruthenate of tetrapropylammonium. Exocoetidae can be treated with hydroxylamine or O-substituted analogue to obtain the corresponding oxime.

Compounds of the invention effective in the treatment of various conditions caused by endoparasites, including, in particular, helminthiasis, which is most often caused by a group of parasitic worms, which are described as Nemat the cattle, and also affects domestic animals and poultry. These compounds are also effective against other nematodes which affect various species of animals, including, for example, Dirofilaria in dogs and various parasites which can infect cattle, accompanying people animals, such as cats and dogs, and people, including gastro-intestinal parasites such as Ancylostoma, Necator, Ascaris, Strongyloides, Trichinella, Capillaria, Toxocara, Toxascaris, Trichuris, Enterobius and parasites which are found in blood or other tissues and organs, such as filaree and extraintestinal stages of Strongyloides and Toxocara, Trichinella.

These compounds are particularly valuable in the treatment of infection by ectoparasites, including, in particular, the infection of arthropod ectoparasites of man, animals and birds, for example, ticks, Ixodes ticks, lice, fleas, padalinimo flies, biting insects and migrating larvae dockrillia insects that infect cattle and horses.

Connections are also insecticides active against household pests such as cockroaches, moths, cojiedo and of houseflies, and suitable against arthropod pests of stored grain and agricultural crops, such as spider mites, aphids, the present invention is characterized as security, and have suddenly very strong systemic activity against winged insects and other important arthropod parasites of cats and dogs.

The compounds of formula I can be introduced in the form of a composition appropriate to the planned route of administration and data types of the animals treated with these compounds and infected by parasites or insects. Connection can be activated by injection subcutaneously or intramuscularly. They can also be administered orally in the form of a capsule, bolus, tablet, chewable tablet or liquid to inject the drug into the oral cavity of the animal, and you can enter them in the form of a composition for local injection or implant. For local injection can be applied composition for immersion, the composition for spraying, powder, dust, composition for pouring onto the animal, a composition for applying stains, fluid spray, shampoo, collar, label or harness. The compositions get the usual manner in accordance with standard veterinary practice. So, for example, capsules, boluses or tablets fashionable to get by mixing the active ingredient with suitable finely divided diluent or carrier, optionally containing disintegr the finished cavity of the animal can be obtained by dispersing the active ingredient in an aqueous solution together with a dispersing or wetting means and injectable preparations can be obtained in the form of a sterile solution or emulsion. Compositions for moulding or coating spots can be obtained by dissolving the active ingredient in a suitable carrier-filler, such as butalbigal, liquid paraffin or non-volatile ether complex, with the possible addition of the volatile component, for example isopropanol. Alternatively preparations for pouring, applying stains or spray on the animal can get kapsulirovaniem to leave the remainder of the active component on the surface of the animal. These compositions will vary according to the weight of active ingredient, depending on the type of processed animal host, the severity and type of infection and the weight of the carrier of the infection. Connections can be entered continuously, in particular for the prevention of known methods. The usual dose for oral, parenteral administration, and the introduction of emptying the animal from about 0.001 to 10 mg per 1 kg of body weight of the animal (as a single dose or as divided the total dose per course of treatment from 1 to 5 days) is sufficient, but there may be cases when there is evidence of higher or lower dose limits, and it is included in the scope of the present invention.

Alternatively the connection can be entered along with food for CNIM animal feed.

For use as an insecticide for pest control connection is applied in the form of a composition for spraying, dust, composition for moulding, emulsions and similar forms in accordance with normal agricultural practice.

Man these compounds is administered in the form of a pharmaceutically suitable preparation in accordance with normal medical practice.

The formation of compounds in accordance with the invention is illustrated by the following examples.

Examples 14-25 are compounds in which R4represents a group capable of either hydrolyzed in vivo.

Example 1. The monosaccharide 22, 23 - dihydroavermectin VA.

22, 23-Dihydroavermectin B1a (50 g) was dissolved in a mixture of isopropanol ( 100 ml) and sulfuric acid ( 1 ml) and stirred at room temperature under nitrogen atmosphere for 48 hours. The reaction mixture was poured on crushed ice and was extracted with dichloromethane ( 2 x 200 ml). The combined extract washed with aqueous saturated sodium bicarbonate solution (100 ml), dried over anhydrous magnesium sulfate and concentrated in vacuum, obtaining white crystals (14 g), which was separated by filtration. The mass spectrum and NMR spectra the m method of 22, 23-dihydroavermectin B1b.

Example 2. The monosaccharide 5-oxo-22, 23-dihydroavermectin VA.

The monosaccharide 22, 23 - dihydroavermectin VA (14 g) was dissolved in diethyl ether (200 ml) and the solution was added activated manganese dioxide (14 g). The mixture was stirred at room temperature for 4 hours, filtered and evaporated to dryness in vacuum, obtaining the title product (11.4 g), NMR-spectrum of which is fully consistent with the proposed structure.

1b - Analog received identical way of monosaccharide 22, 23 - dihydroavermectin 1b.

Example 3. The monosaccharide 5-oximino-22,23-dihydroavermectin VA.

The monosaccharide 5-oxo-22,23-dihydroavermectin B1a (1 g) was dissolved in dry pyridine (25 ml) and the solution was added hydroxylamine hydrochloride (1 g). The reaction mixture under stirring was boiled under reflux for 4 hours and after cooling, was poured on crushed ice and was extracted with dichloromethane (2 x 50 ml). The combined extract was dried over magnesium sulfate and evaporated in vacuum, obtaining the crude resin (1.1 g). This substance was purified liquid chromatography under high pressure column (41,4 x 250 mm, 8 μm, ODS - silica, Rainin, Dynamax commodity EIT the ear, receiving the title product as white solids with so pl. 180 - 190oC. Mass spectrum and NMR spectrum was fully consistent with the proposed structure.

5-Oximino-22, 23 - dihydroavermectin B1b was treated in a similar way from a monosaccharide 5-oxo-22,23-dihydroavermectin B1b.

Example 4. The monosaccharide 22, 23-dihydro-25-cyclohexylaniline B1

25-Cyclohexylaniline B1 (9,9 g) was dissolved in toluene (1 l) was added catalyst of Wilkinson [chloride three (triphenyl-phosphine)rhodium(1)] (a 9.25 g). The solution was first made on a large vibrator Parra (trademark) at room temperature and a hydrogen pressure of 3.5 at. After 3 hours, the pressure in the reaction vessel was discharged, and kept the reaction mixture for 12 hours before adding the next portion of the catalyst (5 g) and was first made as before another 2 hours, after which the reaction mixture remained of the original connection. The solution was filtered, evaporated to dryness in vacuo and the residue was chromatographically on the silicon dioxide, elwira dichloromethane and then a mixture of dichloromethane with methanol (9: 1). The crude product is then again chromatographically on silica (200 g) with elution with a mixture of dichloromethane and methanol (19:1) to give after evaporation rastvorilasj in a mixture of isopropanol (200 ml) and sulfuric acid (2 ml) and the brown solution was stirred at room temperature for 15 hours and then poured into a mixture of ice and water (500 ml) and was extracted with dichloromethane (3 x 200 ml). The organic layer was washed with a saturated aqueous solution of potassium bicarbonate (100 ml), water (2 x 50 ml), dried over anhydrous magnesium sulfate and evaporated in vacuum, obtaining the crude resin, which was chromatographically on silica (100 g) with elution with dichloromethane, then with a mixture of dichloromethane and ethyl acetate (2:1) to give the title compound (8.2 g). The mass spectrum and NMR spectrum is fully consistent with the proposed structure.

Example 5. The monosaccharide 5-oximino-22, 23-dihydro-25-cyclohexylaniline B1

The monosaccharide 22,23 - dihydro-25-cyclohexylaniline B1 (8,2 g) was oxidized in 5-oxoproline using manganese dioxide in anhydrous diethyl ether in accordance with the method of example 2. The crude product was purified by chromatography on silica (50 g) to give 5-exocoetidae (3,22 g) as a yellow foam. It was dissolved in anhydrous pyridine (60 ml) and the solution was added hydroxylamine hydrochloride (3,22 g). After stirring for 15 hours at room temperature was added another portion of hydroxylamine hydrochloride (3,22 g) and the solution was heated at a temperature of up to 50oC as long, until the reaction mixture had no original connection. The solution was poured into water (5 ristoro sodium, was dried over anhydrous sodium sulfate and evaporated to dryness in a vacuum. The crude product was chromatographically on silica (25 g), elwira a mixture of dichloromethane and ethyl acetate (4:1), and finally purified liquid chromatography under high pressure using a column (41,4 x 250 mm, ODS - silica 8 μm, Rainin) Dynamax (trade mark), elwira mixture of methanol and water (9:1) at a rate of 65 ml per minute. Appropriate fractions were combined and evaporated in vacuum, obtaining the title compound (1,53 g). The mass spectrum and NMR spectrum was fully consistent with the proposed structure.

Example 6. The monosaccharide 25-cyclohexylaniline B2

25-Cyclohexylaniline B2 (10 g) is suspended in isopropanol (100 ml) and the suspension was added a solution of sulfuric acid (2 ml) in isopropanol (100 ml). After stirring at room temperature for 24 hours, the clear solution was poured onto ice (600 g) and was extracted with dichloromethane (2 x 100 ml). The organic layer was dried over anhydrous sodium sulfate and evaporated to dryness. The residue was dissolved in carbon tetrachloride and the solution kept at 4oC. the Crystals that stood out slowly, occasionally separated by filtration. Showed that these crystals represented uroy.

Example 7. The monosaccharide 5-oximino-25-cyclohexylaniline B2

The methods of examples 2 and 3 monosaccharide 25-cyclohexylaniline B2 was converted into the title compound. The mass spectrum and NMR spectrum was fully consistent with the proposed structure.

Example 8. The monosaccharide 25-cyclohexylaniline B1

25-Cyclohexylaniline B1 (20 g) was dissolved in tetrahydrofuran (250 ml) and the solution was added a mixture of tetrahydrofuran (250 ml), water (10 ml) and sulfuric acid (10 ml). The mixture was stirred at room temperature for 15 hours and then poured into a mixture of ice (500 g) and water (1 l) and was extracted with dichloromethane (2 x 500 ml). The organic layer was washed with a saturated solution of sodium chloride, dried over anhydrous sodium sulfate and evaporated in a vacuum, getting foamy product. It was chromatographically on silica (150 g) with elution with a mixture of ethyl acetate and dichloromethane (1:1) to give the crude product (13.3 g). Final purification was performed liquid chromatography high resolution (IHVR) with a reversible phase, using a column (41,4 x 250 mm, ODS-silica, 8 μm, Rainin) Dynamax (trade mark). Was suirable mixture of methanol and water (4:1) at a rate of 70 ml per minute, receiving the pure title compound. Masarid 5-oximino-25-cyclohexylaniline B1

The methods of examples 2 and 3 monosaccharide 25-cyclohexylaniline B1 was converted into the title compound. The mass spectrum and NMR spectrum was fully consistent with the proposed structure.

Example 10. The monosaccharide 5-O-tert-butyl-dimethylsilane-25-cyclohexyl-22,23-dihydroavermectin B1

The monosaccharide 22, 23-dihydro-25-cyclohexylaniline B1 (example 4) (12.1 g) and imidazole (7.2 g) was dissolved in dry dimethylformamide (10 ml). In this solution at room temperature was added chloride tert-butyldimethylsilyl (7.9 g). After 18 hours the mixture was poured into a mixture of ice and water (200 ml), acidified to pH 2 using 2N HCl and was extracted with diethyl ether (2 x 80 ml). The combined extract was washed with saturated aqueous sodium bicarbonate solution (50 ml) and water (50 ml), dried over anhydrous sodium sulfate and evaporated in vacuum, obtaining the crude product (14.9 g). The product was further purified by chromatography on silica (kieselgel 60, 230 - 240 mesh, Merck (300 g), elwira a mixture of dichloromethane and ethyl acetate (9:1). Appropriate fractions were combined and evaporated to dryness, obtaining the title product (8,35 g). The NMR spectrum was fully consistent with the proposed structure.

Example 11. 5-Acsaverecord B1a

Avermectin B1a (2.4 g) rivali at room temperature for 18 hours, was filtered and evaporated to dryness in vacuum, obtaining the title product, the NMR spectrum of which is fully consistent with the proposed structure.

Example 12. 5-Examinationin B1a

5-Acsaverecord B1a (800 mg) (example 11) was dissolved in pyridine (10 ml) and the solution was added hydroxylamine hydrochloride (800 mg). After stirring at room temperature for 1 hour the mixture was poured into a mixture of ice (50 g) and water (50 ml), acidified to pH with concentrated hydrochloric acid and was extracted with dichloromethane (3 x 30 ml). The combined extract was washed with water (20 ml), dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure, obtaining the crude product (1 g). The product was chromatographically on the silicon dioxide (kieselgel 60, 230 - 400 mesh, Merck)(100 g), elwira a mixture of dichloromethane and ethyl acetate (2:1), and finally purified liquid chromatography under high pressure using a column (41,4 x 250 mm, ODS-silica 8 μm, Rainin) Dynamax (trade mark) and elwira mixture of methanol and water (85:15) at a rate of 70 ml / min. The desired fractions were combined and evaporated in vacuum, obtaining the title compound (290 mg). The mass spectrum and NMR spectrum was fully consistent with the proposed structure.

P is isopropanol (1 ml) and sulfuric acid (10 ml) and stirred at room temperature under nitrogen atmosphere for 48 hours. Then was added a saturated aqueous solution of sodium bicarbonate (1 ml) and the product was extracted with ethyl acetate (2 x 5 ml). The combined extract was dried over anhydrous magnesium sulfate and concentrated in vacuum. The crude product (25 mg) was purified liquid chromatography high-pressure column (24 x 250 mm, ODS-silica 5 μm, Beckman) Ultrasphere (trade mark), elwira mixture of methanol and water (85:15) at a rate of 20 ml per minute. Appropriate fractions were combined, receiving the title product. The mass spectrum and NMR spectrum was fully consistent with the proposed structure.

Example 14. The monosaccharide 5-(trimethylacetamido-25-cyclohexyl-22,23-dihydroavermectin B1

In a mixed solution of monosaccharide 5-oximino-25-cyclohexyl-22, 23-dihydroavermectin B1 (example 5) (50 mg) in dichloromethane (2 ml) at room temperature was added triethylamine (72 ml), then chloride trimethylacetyl (80 ml). After keeping the mixture for 18 hours was added an aqueous citric acid solution (10 wt./vol.% 2 ml). The organic layer was separated, washed with saturated aqueous sodium chloride (2 ml), dried over anhydrous sodium sulfate and evaporated to dryness in vacuum, obtaining the crude product, the cat is e fractions were combined and evaporated to dryness in a vacuum, receiving the product (53 mg), which was further purified liquid chromatography high-pressure column (21.2 x 250 mm, ODS-silica, 5 μm, Rainin, Dynamax (trade mark), elwira rate of 20 ml / min with a mixture of methanol and water (95:5). Appropriate fractions were combined and evaporated in vacuum, obtaining the title compound as a white powder (18 mg). The mass spectrum and NMR spectrum was fully consistent with the proposed structure.

Example 15. The monosaccharide 5-(benzoylamino)-25-cyclohexyl-22, 23-dihydroavermectin B1

The monosaccharide 5-oximino-25-cyclohexyl-22, 23-dihydroavermectin B1 (example 5) (70 mg) in dichloromethane (30 ml) was treated with triethylamine (50 ml) and benzyl chloride (100 ml) and the target product was extracted by a method identical to that described in example 42. Purification was performed liquid chromatography high-pressure column (41,4 x 250 mm, ODS - silica 8 μm, Rainin) Dynamax (trade mark), elwira at a speed of 45 ml / min with a mixture of methanol and water (90:10). Appropriate fractions were combined and evaporated in vacuum, obtaining the title compound as a white powder (28 mg). The mass spectrum and NMR spectrum was fully consistent with the proposed structure.

Example 16. The monosaccharide 5-(N-methylimino-25-cyclohexyl - 22, 23-dihydroavermectin B1 (example 5) (106 mg) in dichloromethane (10 ml) was added methyl isocyanate (15 ml) and the mixture was stirred for 1 hour. Then add an additional amount of methyl isocyanate (30 ml) and the reaction mixture was stirred for 72 hours before adding saturated aqueous solution of sodium chloride (10 ml) and diethyl ether (30 ml). The organic extract was dried over anhydrous sodium sulfate and evaporated to dryness in vacuum, obtaining the crude product (150 mg) which was purified liquid chromatography high-pressure column (41,4 x 250 mm, ODS-silica, 8 μm, Rainin)Dynamax (trade mark), elwira at a speed of 45 ml / min with a mixture of methanol and water (91:9). Appropriate fractions were combined and evaporated to dryness in vacuum, obtaining the title compound as a white powder (80 mg). The mass spectrum and NMR spectrum was fully consistent with the proposed structure.

Example 17. The monosaccharide 5-(N, N-dimethylcarbamoyl)-25-cyclohexyl-22, 23-dihydroavermectin B1

In a mixed solution of monosaccharide 5-oximino-25-cyclohexyl-22, 23-dihydroavermectin B1 (example 5) (50 mg) in dichloromethane (2 ml) at room temperature was added triethylamine (72 ml) and 4-dimethylaminopyridine (1 ml), then chloride is (58 ml) and the reaction mixture was stirred for 18 hours. Then added an aqueous citric acid solution (10 wt./vol.%, 2 ml) and diethyl ether (20 ml). The organic layer was separated, washed with saturated aqueous sodium chloride (5 ml), dried over anhydrous sodium sulfate and evaporated to dryness in vacuum, obtaining the crude product, which was purified liquid chromatography high-pressure column (21.2 x 250 mm, ODS-silica, 5 μm, Rainin) Dynamax (trade mark), elwira at a rate of 10 ml / min with a mixture of methanol and water (90:10). The desired fractions were combined and evaporated in vacuum, obtaining the title compound as a white powder (18 mg). The mass spectrum and NMR spectrum was fully consistent with the proposed structure.

Example 18. The monosaccharide 5-(4-methylpiperazine-1 carbonyloxy)-25-cyclohexyl-22, 23-dihydroavermectin B1

In a mixed solution of N-methylpiperazine (0,65 ml) and triethylamine (1.3 ml) in toluene (25 ml) at 0oC was added dropwise a solution of phosgene in toluene (20%, 5,1 ml) for 15 minutes. The reaction mixture was stirred to warm to room temperature, stirred for 3 hours, filtered and concentrated to approximately 10 ml under reduced pressure, obtaining a solution of 1-chlorocarbonyl-4-methylpiperazine that education is) and 4-dimethylaminopyridine (5 mg) in dichloromethane (10 ml) at room temperature in accordance with the method, described in example 45. Purification of the product was performed by chromatography on silica (kieselgel 60, 230-400 mesh mesh, Merck) (35 g), elwira dichloromethane. The desired fractions were combined and evaporated in vacuum, obtaining the product (53 mg), which was further purified liquid chromatography high-pressure column (21.1 × 250 mm, ODS-silica, 5 μm, Rainin) Dynamax (trade mark), elwira rate of 20 ml / min with a mixture of methanol and water (95:5). The desired fractions were combined and evaporated in vacuum, obtaining the title compound as a white powder. The mass spectrum and NMR spectrum was fully consistent with the proposed structure.

Example 19. The monosaccharide 5-(tert-butyloxycarbonyl(-25-cyclohexyl-22, 23-dihydroavermectin B1

In a mixed solution of monosaccharide 5-oximino-25-cyclohexyl-22, 23-dihydroavermectin B1 (example 5) (60 mg) and triethylamine (50 mg) in dichloromethane (5 ml) at room temperature was added tert-butyl ester of carbonic acid (60 mg). After conditioning for 48 hours the reaction mixture is evaporated to dryness in vacuum, obtaining a residue that was dissolved in dichloromethane and the solution was chromatographically on the silicon dioxide (kieselgel 60, 230-400 mesh mesh, Merck) (5 g), elwira dichloromethane. Suitable ACC-spectrum and the NMR spectrum was fully consistent with the proposed structure.

Example 20. The monosaccharide 5-(N-(4-formylphenyl)-carbamoyloximes)-25-cyclohexyl-22, 23-dihydroavermectin B1

4-formylphenylboronic obtained in accordance with the procedure described in J. Med. Chem., 32(10), 2354 (1989), processed monosaccharide 5-oximino-25-cyclohexyl-22, 23-dihydroavermectin B1 (example 5) (500 mg) in dry dichloromethane (50 ml) at room temperature for 1 hour in accordance with the method described in example 15. Purification of the target product was carried out on silica (kieselgel 60, 230 - 400 mesh, Merck) (125 g), elwira with a mixture of hexane and diethyl ether with a gradient from 1:1 to 20:80. The desired fractions were combined and evaporated to dryness in vacuum, obtaining the title compound as a white powder (300 mg). The mass spectrum and NMR spectrum was fully consistent with the proposed structure.

Example 21. The monosaccharide 5-(N-(4-diethylaminomethyl)phenyl)carbamoyloximes)-25-cyclohexyl-22, 23-dihydroavermectin B1

Chloride 4-Diethylaminoethanol received in accordance with the methodology described in published U.S. patent US-4623486, were treated with monosaccharide 5-oximino-25-cyclohexyl-22, 23-dihydroavermectin B1 (example 5) (100 mg) in dry dichloromethane (50 ml) containing triethylamine (450 ml) and 4-dimethylaminopyridine (126 new product was carried out on silica (kieselgel 60, 230-400 mesh mesh, Merck) (5 g), elwira mixture of methanol and dichloromethane with gradient 0:100 to 10:90. Appropriate fractions were combined and evaporated to dryness in vacuum, obtaining the title compound as a white powder (11 mg). The mass spectrum and NMR spectrum was fully consistent with the proposed structure.

Example 22. The monosaccharide 5-(N-(4-(4-methyl-1-piperazinylmethyl)phenyl)-carbamoyloximes) -25-cyclohexyl-22, 23-dihydroavermectin B1

Chloride 4-(4-methylpiperazin-1-ylmethyl)benzoyl obtained in accordance with the technique described in patent publication U.S. US-4623486, were treated with monosaccharide 5-oximino-25-cyclohexyl-22, 23-dihydroavermectin B1 (example 5) by the method identical to that described in example 20. The title compound was obtained as a white powder (18 mg). The mass spectrum and NMR spectrum was fully consistent with the proposed structure.

Example 23. The monosaccharide 5-(N-(3-pyridyl-carbonyl)carbamoyloximes)-25-cyclohexyl-22, 23-dihydroavermectin B1

In a mixed solution of nicotinamide (4,88 g) in dry 1,2-dichloroethane (500 ml) was added dropwise chloride, oxalyl (5,24 ml). The mixture was boiled under reflux for 4.5 hours, then cooled and filtered. The resulting solution containing nicoti mg) in dichloromethane (10 ml) at room temperature. After standing the reaction mixture for 18 hours was added an additional amount of solution nicotinereplacement (25 ml) and the mixture was stirred at room temperature for another 18 hours. The mixture is then evaporated to dryness in vacuum, obtaining a residue that was purified liquid chromatography high-pressure column (41,4 x 250 mm, ODS-silica, 8 μm, Rainin) Dynomax (trade mark), elwira at a speed of 45 ml / min with a mixture of methanol, acetonitrile and water (20:65:15). The desired fractions were combined and evaporated in vacuum, obtaining the title compound as a white powder. The mass spectrum and NMR spectrum was fully consistent with the proposed structure.

Example 24. The monosaccharide 5-(N-(3-pyridyl)-carbamoyloximes)-25-cyclohexyl-22, 23-dihydroavermectin B1

In a solution of dihydrochloride of nicotinic acid hydrazide (2 g) in water (10 ml) was added a solution of sodium nitrite (1.6 g) in water (10 ml), keeping the temperature below 20oC. Then was added diethyl ether (50 ml) and the mixture was podslushivaet careful addition of solid sodium bicarbonate. The organic layer was separated, washed with water (20 ml), dried over anhydrous magnesium sulfate and evaporated to dryness in a vacuum, getting nicotinate (1.1 g) with so pl. 54oC., receiving a solution containing 3-predilatation. Part of this solution (1 ml) was treated with a monosaccharide 5-oximino-25-cyclohexyl-22, 23-dihydroavermectin B1 ( example 5) (100 mg) in toluene (10 ml) at room temperature for 1 hour, then the mixture was poured into a mixture of diethyl ether and water (1:1, 30 ml). The organic layer was separated, dried over anhydrous magnesium sulfate and evaporated to dryness in vacuum, obtaining a residue (130 mg) which was purified liquid chromatography high-pressure column (41,4 x 250 mm, ODS-silica, 8 μm, Rainin) Dynamax (trade mark), elwira at a speed of 45 ml / min with a mixture of methanol and water with a ratio of 85:15, which was changed after 15 minutes, until the ratio of 87:13. Appropriate fractions were combined and evaporated in vacuum, obtaining the title compound as a white powder (52 mg). The mass spectrum and NMR spectrum was fully consistent with the proposed structure.

Example 25. The monosaccharide 5-(arylcarbamoyl-oximino)-25-cyclohexyl-22, 23-dihydroavermectin B1

Reaction of monosaccharide 5-oximino-25-cyclohexyl-22, 23-dihydroavermectin B1 (example 5) (500 mg) with arylisocyanates (108 mg) in dichloromethane (50 ml) according to the method described in example 15, was obtained the title compound in VI is P CLASS="ptx2">

Example 26. Obtaining veterinary compositions for oral administration.

Any of the data obtained in the above examples, the product is mixed with lactose powder and fill with a mixture of hard gelatin capsules based 1.0 mg of active compound per capsule.

Example 27. Activity of the compounds of the present invention against fleas.

1. Test in vitro by determining the activity of compounds against fleas Ctenocephalides felis is conducted according to the following General method.

1.1. Test compounds dissolved in DMSO to obtain a working solution with a concentration of 0.2 mg/ml Aliquot of 25 ml add 5 ml of a mixture of cow blood and citrate with obtaining the initial concentration for test 1 µg/ml. Make dilution in DMSO and 25 μl of the diluted solution is added to 5 ml of a mixture of cow blood and citrate, as described above, receiving concentrations for testing 0.5 μg/ml, 0.25 microgram/ml, etc.

1.2. A mixture of citrate and blood of a calf prepared by adding 100 ml of citrate solution (22 g of sodium citrate, 8.0 g of citric acid monohydrate, 24, 5 g of glucose in 100 ml of water) to 500 ml of blood of a calf.

1.3 20-25 adult fleas Ctenocephalides felis (cat flea) collect and poseshayushie.

1.4. 5 ml of a mixture of cow blood and citrate containing the test compound, is placed in a glass container, the temperature at which the support 37oC using a water outer jacket. Pull paraffin film over the open top of the container, obtaining a dense membrane, through which fleas can eat. The camera for testing, containing fleas, carefully placed on the parafilm membrane, and then the tank/chamber for testing overturn in order to bring the blood into contact with the membrane and fleas began to eat.

1.5. The fleas were allowed to eat for 6 hours, after which the camera for testing removed and stored over night at 25oC in a humid atmosphere.

1.6 See fleas that have fallen and/or died, and fix their percentage. Compounds active at concentration 1 μg/ml, further testing at lower doses. When compounds were tested more than once (n>1), record the total percentage. The results obtained are shown in the attached table.

2. Testing in vitro

2. The potency of the compound of example 5 of the present invention (IR-124, 114) was also determined in vivo in mice.

The test is carried out in 10 hungry adult fleas, which were isolated from eating blood, placed in sterile tubes.

2.2. Test compounds administered orally to DM1 mice weighing 20 g, which is then incubated for 2-4 hours, anaesthetize and placed in tubes with fleas.

2.3. Fleas are in contact with mice within 1 hour.

2.4. Then the mice are removed from the tubes and full of fleas removed from mice and placed in tubes containing long, narrow strip of paper, and leave it there. The end of the tube is closed by a grid with very small holes.

2.5. Examine the number of fleas that were paralyzed and/or dead after 4, 24 and 48 hours after dosing, and record the number of deaths or survivors of individuals.

2.6. It was found that the compound of example 5 (IR 124, 114) is active in this test at a dose of 3 mg/kg, with no signs of any toxicity was not observed.

1. Derived embryo death of General formula I

< / BR>
where the dashed line in 22(23)-position denotes an optional carbon-carbon bond, and this bond is present and R1missing or this relationship is absent and R1represents H or HE;

R2pri H or HE;

R4represents H or a group capable of either hydrolyzed in vivo to form compounds in which R4represents H, and which represents acetyl, tert-butylcarbamoyl, tert-butoxycarbonyl, benzoyl, methylpiperidine, N-methylcarbamoyl, N, N-dimethylcarbamoyl, formylphenylboronic, N-(4-diethylamino-were)carbarnoyl, N-(4-methyl-1-piperazineethanol) carbarnoyl, N-(3-pyridylcarbonyl)carbarnoyl, N-(3-pyridyl)carbarnoyl or arylcarbamoyl;

R5is a HE.

2. Connection on p. 1, in which R2represents cyclohexyl.

3. Connection under item 1 or 2, in which the optional bond in 22(23)-position is absent and R1represents N.

4. Connection on p. 1, chosen from:

the monosaccharide 5-oximino-22,23-dihydroavermectin B1a,

the monosaccharide 5-oximino-22,23-dihydro-25-cyclohexylaniline B1,

the monosaccharide 5-oximino-25-cyclohexylaniline B2,

the monosaccharide 5-oximino-25-cyclohexylaniline B1,

the monosaccharide 5-oximino-embryo death WA.

5. Connection on p. 1, chosen from:

the monosaccharide 5-(trimethylacetamido)-25-cyclohexyl-22,23-dihydroavermectin B1,

a monosaccharide is clohessy-22,23-dihydroavermectin B1,

the monosaccharide 5-(N, N-dimethylcarbamoyl)-25-cyclohexyl-22,23-dihydroavermectin B1,

the monosaccharide 5-(4-methylpiperazine-1 carbonyloxy)-25-cyclohexyl-22,23-dihydroavermectin B1,

the monosaccharide 5-(tert-butyloxycarbonyl)-25-cyclohexyl-22,23-dihydroavermectin B1,

the monosaccharide 5-(N-(4-formylphenyl)carbamoyloximes)-25-cyclohexyl-22,23-dihydroavermectin B1,

the monosaccharide 5-(N-(4-diethylaminomethyl)phenyl)carbamoyloximes)-25-cyclohexyl-22,23-dihydroavermectin B1,

the monosaccharide 5-(N-(4-methyl-1-piperazinylmethyl)phenyl)carbamoyloximes)-25-cyclohexyl-22,23-dihydroavermectin B1,

the monosaccharide 5-(N-3-pyridylcarbonyl)carbamoyloximes-25-cyclohexyl-22,23-dihydroavermectin B1,

the monosaccharide 5-(N-(3-pyridyl)carbamoyloximes)-25-cyclohexyl-22,23-dihydroavermectin B1,

the monosaccharide 5-(N-arylcarboxamide)-25-cyclohexyl-22,23-dihydroavermectin B1.

6. Pharmaceutical or veterinary composition having anti-parasitic activity, containing derivatives of avermectins as an active agent and a pharmaceutically suitable carrier or excipient, characterized in that it contains as active agent a compound of the formula I according to any one of paragraphs.1 - 5 th agent to combat fleas.

8. The method of obtaining the compounds of formula I under item 1

< / BR>
where R1, R2, R3, R4and R5take the values specified above,

which involves the following stages:

(i) oxidation of compounds of formula II

< / BR>
where the dotted line, R1, R2and R3have the above values;

R5has the above values or R5represents alejandroisaacura,

obtaining the compounds of formula III

< / BR>
(ii) reaction of the compound of formula III with the compound of the formula

R4-O - NH2,

where R4has the above meanings;

R5represents alejandroisaacura,

and hydrolysis of the compounds in the compound of formula I,

(iii) if necessary, replacement of the group R4when she represents H, a group capable of either hydrolyzed in vivo to form compounds in which R4represents H,

and, if necessary,

(iv) hydrogenation of compounds to restore the double bond in position 22(23) in a single communication.

 

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