Derivatives doramectin, methods for their preparation, methods of treatment

 

(57) Abstract:

Derivatives doramectin formula III, where R1- (C2- C5) alkanoyl or phenoxyacetyl, optionally substituted by 1 to 3 ( C1- C4) alkyl, ( C1- C4) alkoxy or halogen; P2is hydrogen or phenoxythiocarbonyl, optionally substituted by 1 to 3 ( C1- C4) alkyl, ( C1- C4) alkoxy or halogen. Doramectin and intermediates obtained by the method according to this invention, can also be used as antiparasitics agents. 7 C. and 16 h.p. f-crystals.

The present invention relates to new intermediates in obtaining the compounds of the following formula (I)

< / BR>
hereinafter doramectin, and an improved process for the preparation of these intermediates. The invention also relates to the use of these intermediates as antiparasitics compounds. Further, the invention relates to an improved method of obtaining doramectin from compounds of the following formula (II).

< / BR>
Doramectin - antiparasitically and antihelminthic agent wide spectrum of action belonging to the class of secondary metabolites, known as Avermes is avermectin, such as ATCC 31267, 31271 or 31272 under aerobic conditions in an aqueous nutrient medium containing inorganic salts and assimilable sources of carbon and nitrogen, as described in U.S. patent N 5089480 that there is a link. Another strain of microorganism producing avermectin is Streptomyces avermitilis ATCC 53568, which is described by Dutton with TCS. (Dutton et al., Jornal of Antibiotics, 44, 357-65 (1991). Streptomyces avermitilis ATCC 31267, and 31271 31272 deposited in accordance with the terms of the Budapest agreement in the American Type Culture Collection, 12301 Parklawn Drive, Rockvill, Maryland 20852, USA.

During the process of fermentation, which gives the compound of the above formula (I) can be allocated to different products. The main by-product, which can be selected during fermentation defined above microorganisms, has the structure above described formula (II). The selection of the compounds of formula (II) from a fermentation broth Streptomyced avermitilis ATCC 53568 described by Dutton with employees (see above). The compound of formula (II) is an active antiparasitics and anthelminthic compound that has been described and protected in U.S. patent N 5089480. However, doramectin is preferred. The method according to the present invention allows the use of side products is>The total change in this method is the elimination of the hydroxy-group on the C atom-23 education in the molecule olefinic linkages between positions C-22 and C-23. Earlier it was reported about the method such as turning slightly different avermectins, which are characterized by low output. For example, if the atom C-25 instead of second-butilkoi group is tsiklogeksilnogo, then the transformation is achieved as the result of five sequential stages with a total output of approximately 3.6% (Mrozic et al. , Tetrahedron Letters, 1982, 23, 2377-78). Due to the low output in this way it is desirable to invent a way in which the source material more effectively would be turned to doramectin.

Therefore, the aim of the present invention is the conversion of the compounds of formula (II) in doramectin with a high total yield. A further purpose of this invention to provide useful intermediates for the method.

The present invention provides for obtaining new intermediates of the formula (III)

< / BR>
where

R1is (C2-C5)alkanoyl or aryloxyacetic, and

R2is hydrogen or aryloxyalkyl.

This invention further provides a receiving new intermediates of the formula (IV)1is (C2-C5)alkanoyl, phenoxyacetyl or (C1-C4)alkylphenoxy, and R2is hydrogen or (C1-C4)alkylenediamines. Particularly preferred compounds within this group are the compounds of formula (III), where R1is acetyl or phenoxyacetyl, and R2is hydrogen or p-tolylacetylene.

Also preferred compounds according to this invention are the compounds of formula (IV), where R1is (C2-C5)alkanoyl, phenoxyacetyl or (C1-C4)alkylperoxyl. Particularly preferred compounds within this group are the compounds of formula (IV), where R1is acetyl or phenoxyacetyl.

This invention further provides a method of obtaining

compounds of formula (I)

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includes successive stages:

(a) interaction of the compounds of formula (II)

< / BR>
with allermuir agent of the formula (R4CO)2O or R4COX, where R4is relaxometer and X is Cl or Br, with the formation of the compounds of formula (III), where R1is aryloxyacetic and R2is H;

(b) interaction of the compounds of the formula (III), where R1is aryloxyacetic and R2- this is s (III), where R1is aryloxyacetic and R2is aryloxyalkyl;

(C) interaction of the compounds of the formula (III), where R1is aryloxyacetic and R2is aryloxyalkyl, in an inert solvent at a temperature of from 150oC to 200oC for from 2 to 48 hours with calcium carbonate when the continuous supply of inert gas with the formation of the compounds of formula (IV), where R1is aryloxyacetic; and

(g) interaction of the compounds of the formula (IV), where R1is aryloxyacetic with base in alcohol with the formation of the compounds of formula (I).

In particular, the preferred method in this method is the one where the inert gas is nitrogen. A particularly preferred method in this proposed method is one where the base is NH3, KOH, KCN, Na2CO3or NaOAc. Even more preferred of ways in the person of the preferred method are those which are particularly preferred compounds according to this invention.

The invention is further directed to a method of obtaining the compounds of formula (I), including successive stages:

(a) interactions of the compounds of formula (II) with allermuir agent of the formula (R4CO)1is (C1-C4)alkanoyl and R2is H;

(b) interaction of the compounds of the formula (III), where R1is (C1-C4)alkanoyl) and R2is H, with a compound of formula R3OC(=S)X, where R3is aryl and X is Cl or Br, with the formation of the compounds of formula (III), where R1is (C1-C4)alkanoyl and R2is aryloxyalkyl;

(C) the interaction of the compounds of the formula (III), where R1is (C1-C4)alkanoyl and R2is arylacetylenes, in an inert solvent at a temperature of from 150oC to 200oC for from 2 to 48 hours with calcium carbonate when the continuous supply of inert gas with the formation of the compounds of formula (IV), where R1is (C1-C4)alkanoyl; and

(g) interaction of the compounds of the formula (IV), where R1is (C1-C4)alkanoyl with lithium aluminum hydride, cyanoborohydride sodium or triethylborohydride lithium in an inert solvent with the formation of the compounds of formula (I).

In particular, the preferred method in this method is the one where the inert gas is nitrogen. Even more preferred ways in particularly preferred methods are those in which the alley provides a method of treatment of mammals from parasitic diseases, involving the introduction of a specified mammal antiparasitics effective amount of the compounds of formula (II) or (IV).

Compounds according to this invention can easily receive in accordance with the method of the present invention, which are described below.

The compound of formula (I) in this invention is doramectin useful antiparasitically and antihelminthic agent, which is described in U.S. patent N 5089480, to which there are links. Doramectin according to this invention are obtained from compounds of the above formula (IV) interaction of the compounds of the formula (IV) with lithium aluminum hydride, cyanoborohydride sodium or triethylborohydride lithium or saponification of esters depending on the nature split the air.

If R2is (C2-C5)alkanoyl, (C2-C5)alkanoyloxy transformed into the hydroxy-group by reacting the compounds of formula (IV) where R1is (C2-C5)alkanoyl, with lithium aluminum hydride, cyanoborohydride sodium or triethylborohydride lithium in an inert solvent at a temperature from -100oC and 0oC for from 15 minutes to 24 hours. Use inert solvents for this reaction, diethyl ether, dioxane, tetrahydrofuran, 2-methoxymethyl ether and 1,2-dimethoxyethane. Particularly preferred regenerating agent for this reaction is triethylborohydride lithium, and especially good solvent is tetrahydrofuran. In General, the temperature during mixing of the reagent with the substrate and within a short time after this, usually from 15 minutes to 1 hour, supported from -78oC to -70oC. Immediately after mixing, usually within 15 minutes to 1 hour, the temperature is slowly increased to room. The reaction mixture was quenched and the product of formula (I) was isolated using well known to the average person skilled in the art standard methods.

If R1is aryloxyacetic, alloctype transformed into the hydroxy-group by reacting the compounds of formula (IV) where R1is aryloxyacetic, with alkali metal hydroxide in alcohol at a temperature of from -75oC and 0oC for from 15 minutes to 24 hours. Particularly preferred alkali metal hydroxide is potassium hydroxide and a particularly preferred solvent is methanol. In General, the reaction is carried out at a temperature of -35oC for about 1 hour, p is Antartic ways. Alternatively, the reaction is carried out by replacing the alkali metal hydroxide ammonia. In the case of ammonia, the reaction is in General carried out at temperatures from -35oC and 0oC for 1 to 16 hours. The reaction is preferably carried out in methanol at -15oC for 5-6 hours. The product of formula (I) was isolated using standard methods of organic chemistry well known to the average expert in the field.

The compound of formula (IV) according to this invention can easily receive in accordance with the method of this invention by the interaction of the compounds of formula (III) according to this invention in a high-boiling solvent, such as 2-methoxyethylamine ether or 2-ethoxyethyl ether, but not limited to, at temperatures from 150oC to 200oC for from 2 to 48 hours. To achieve maximum yield in the reaction mixture is continuously serves inert gas, such as nitrogen or argon. In particular, it is preferable to conduct the reaction to 156o-158oC 2-methoxyethanol the air with continuous transmission of nitrogen for 12 hours. Upon completion of the reaction product of the formula (IV) was isolated using well known to the average person skilled in the art standard methods. according to the method according to this invention by reacting the compounds of formula (III) according to this invention, where R2is hydrogen, halogenation formula R3OC(=S)X, where R3defined above, in an inert solvent in the presence of a proton acceptor, such as pyridine or 4-dimethylaminopyridine, for from 30 minutes to 12 hours. Suitable inert solvents for this reaction include ethyl acetate, 1,2-dimethoxyethane, 2-methoxymethyl ether, 2-methoxyethylamine ether, aromatic solvents such as toluene, xylene and benzene, and chlorinated solvents such as chloroform and methylene chloride. Preferred solvents for this reaction include ethyl acetate or toluene. It is preferable to conduct the reaction for 2 to 4 hours. The reaction mixture is heated to a temperature of from 40oC to the boiling temperature selected for the reaction solvent. The reaction mixture was quenched and the product of formula (III), where R2is arylacetylenes, isolated in accordance with standard methods.

The compounds of formula (III) according to this invention, where R2is hydrogen and R1is (C2-C5)alkanoyl or aryloxyacetic, easy to get in accordance with the method of the present invention. The compound of formula (II) reacts with allermuir agent in an inert rastvoriteli, morpholine or di(ISO-propyl)ethylamine, but not limited to, at temperatures from -75oC and 0oC for from 5 minutes to 8 hours. Suitable inert solvents for this reaction include aromatic solvents such as toluene, benzene or xylene, or chlorinated solvents such as methylene chloride, chloroform or 1,2-dichloroethane. Suitable allerease agents are the acid halides, typically the acid chlorides, and anhydrides of the acids. If Alliluyeva agent is used, the acid chloride of the acid, particularly preferred organic amine is pyridine. If Alliluyeva agent uses the anhydride of the acid, particularly preferred organic amine is triethylamine. In General, the preferred solvents for these reactions, in which as Alliluyeva agent is used, the acid chloride or acid anhydride, are chlorinated solvents. In particular, preferred is methylene chloride. Upon completion of the reaction, the reaction mixture was quenched and the product of formula (III), where R2is hydrogen and R1is (C2-C5)alkanoyl or aryloxyacetic, isolated using standard methods of organic x the clients avermectin, such as ATSS 31267, 31271 or 31272, as described in U.S. patent N 5089480, to which there are links. Other methods for obtaining compounds of formula (II), including the allocation of fermentation broth of Streptomyces avermitilis ADS 53568 described by Dutton employees (Dutton et al., Journal of Antibiotics, 44, 357-65 (I991).

New compounds (III)-(IV) of the present invention are used as intermediates in the synthesis of the compounds of formula (I), doramectin, from compounds of formula (II).

New compounds (III)-(IV) of the present invention are also used as antiparasitics agents. The applicability of these compounds (III)-(IV) as antiparasitics agents demonstrated the activity of these compounds in model experiments in vivo in rodents, as described by Grahanam with employees (Gration et al., Veterinary Parasitology, 42, 1992, 273-279.

Compounds of formulas (III) and (IV) are highly active antiparasiticsthe agents having the application, in particular, as a sedative, anti-ectoparasites, insecticides and acaricides (anti-mites).

Compounds of formulas (III) and (IV) effective for the treatment and prevention of and variety of cases caused by endoparasites, including, in particular, helminthes, Kolesnik economic losses in pig production sheep breeding, horse breeding and animal husbandry in General, as well as to the defeat of domestic animals and poultry. Doramectin also effective against other nematodes which affect various species of animals, including, for example, Dirofilaria in dogs, and against various parasites such as Ancylostoma, Necator, Ascaris, Strongyloides, Trichinella, Capillaria, Trichuris, Enterobius and against other parasites found in blood or other tissues and organs, such as filiform worms, and against Strongyloides and Trichinella in extraintestinal stages.

Compounds of formulas (III) and (IV) are also valuable in the treatment and prevention of an ectoparasitic infections, including, in particular, Arthropoda ectoparasites of animals and birds such as ticks, lice, fleas, meat flies, biting insects and migrating larvae of Diptera, which can affect cattle and horses.

Compounds of formulas (III) and (IV) also exhibit insecticidal activity against parasites of dwellings, such as cockroaches, clothes moth, carpet beetle and fly home, as well as against insects in grain and agricultural plants, such as spiders, hafidi, caterpillars, and against migratory Orthoptera, such as the locust.

Compounds of formulas (III) and (IV) are introduced in the form of compositions in accordance with IV". When used as a sedative compound may be administered orally as capsules, pastilles, tablets, or preferably in the form of a liquid composition or, alternatively, may be in the form of injection, infusion or in the form of the implant. Such compositions are prepared in the usual manner in accordance with standard practice in veterinary medicine. So, capsules, pellets, or tablets are prepared by mixing the active ingredient with suitable finely diluent or carrier, optionally containing crushed agent and/or binder such as starch, lactose, talc, magnesium stearate, etc., a Liquid composition is prepared by dispersing the active ingredient in an aqueous solution together with dispersing agents or wetting, etc. and composition for injection is prepared in the form of a sterile solution, which may contain other substances, for example enough salts or glucose to make the solution property isotonicity with blood. These formulations can vary in weight of active compound, depending on the species of the host being treated, the severity and type of infection and the body weight of the host. In the General case, for oral administration or injection of a dose of approximately from 0.001 enough, but there may be circumstances in which shown a higher or lower dose, and such are within the scope of this invention. For oral administration, or injections, the most preferred dose of from 0.02 to 2 mg/kg of body weight of the animal as a single dose or in separate doses within 1 to 5 days.

Alternatively, the compounds can be administered to animal feed; for this purpose, prepare a concentrated feed additive or admixture for mixing with the normal animal feed.

For use as an insecticide and processing of agricultural plant compounds used in the form of aerosols, powders, emulsions and the like in accordance with standard practice in agriculture.

As used above and in the attached claims, the term "inert solvent" refers to any solvent which does not react with the starting materials, reagents, intermediates or products in any way, adversely affecting the yield of the target product. The term "inert gas" refers to any gas that does not react with the starting materials, reagents, intermediates or products in any way, negatively affecting the output Clevo is or phenyl, arbitrarily substituted with one, two or three (C1-C4)CNS groups or halogen atoms.

As used above and in the attached claims, the term "aryloxy" means phenyloxy or fenoxaprop optionally substituted one, two or three (C1-C4)alkyl, (C1-C4)CNS groups or halogen atoms.

The present invention is illustrated in the following examples. However, it should be understood that the invention is not limited to the specific details of these examples.

Example 1. The compound of formula (III), where R1is phenoxyacetyl and R2is hydrogen

A solution of 20 g of compound of formula (II) in 400 ml of methylene chloride and 10 ml of anhydrous pyridine is cooled by passing nitrogen to -78oC, and within 10 minutes are added dropwise to 15.1 ml phenoxyacetamide. The reaction mixture was kept at a temperature of from -70oC to -78oC for 1.5 hours, then quenched with 250 ml saturated sodium bicarbonate solution. The mixture is stirred for 1.5 hours, the organic layer is separated, re-extracted with 250 ml saturated sodium bicarbonate solution and dried over anhydrous MgSO4. The solvent is removed in vacuun 7.9 minutes (HPLC, Ultrasphere ODS 5, 25 cm x 4.6 mm (Beckman); methanol:acetonitrile: water 559:383:58; 1.5 ml/min; UV 245 nm).

Example 2. The compound of formula (III), where R1is phenoxyacetyl and R2is p-tolylacetylene

A solution of 10 g of compound indicated in the heading of example 1, in 100 ml of toluene and 30 ml of anhydrous pyridine is heated by passing nitrogen to 100oC and within 5 minutes was added 7.7 ml O-p-tailgatenation. The reaction mixture was kept at 1005oC for 2 hours to complete the reaction, cooled to approximately the 50oC and the solvents removed in vacuum with the formation of a brown precipitate. The residue is dissolved in 120 ml of toluene, the solution is extracted with 120 ml of water and twice with 100 ml saturated sodium bicarbonate solution. Separate the organic layer evaporated to 80 ml, filtered and chromatographic using system Prep-500 (Waters Associates). Eluent composition has hexane: ethyl acetate 75: 25. The fractions containing the product are pooled, the solvent is removed in vacuum, get 9,8 g 95,7% of the product as a yellowish brown solid (yield 83,2%). The retention time of 15.2 minutes (HPLC, Ultrasphere ODS 5, 25 cm x 4.6 mm (Beckman); methanol:acetonitrile:water 559:383:58; 1.5 ml/min; UV 245 nm).

Example 3. 5,4"-O-diphenoxyethane (E of example 2, in 90 ml of 2-methoxyethanol ether add 4.5 g of calcium carbonate. Then the mixture is heated with good stirring and continuous transmitting current of nitrogen to 156o-158oC. After keeping at 156o-158oC within 24 hours analysis of the mixture shows the presence of 6.68 g of product (yield 90%). After filtering the mixture, the solvent is removed in vacuum; receive the product as a yellow resin. Retention time of 10.4 minutes (HPLC, Ultrasphere ODS 5, 25 cm x 4.6 mm (Beckman); methanol:acetonitrile:water 559:383:58; 1.5 ml/min; UV 245 nm).

Example 4. 5,4"-O-Diphenoxylate-doramectin (compound of formula (IV), where R1is phenoxyacetyl)

To a solution of 1 g of compound indicated in the heading of example 2 in 20 ml of 2-ethoxyethyl ether is added 500 mg of calcium carbonate. Then the mixture is heated with good stirring and continuous transmitting current of nitrogen to 185oC. After keeping at 185oC for 5 hours, the reaction mixture was cooled to room temperature and filtered. Analysis of the solution by HPLC (Ultrasphere ODS 5, 25 cm x 4.6 mm (Beckman); methanol:acetonitrile:water 559: 383: 58; 1.5 ml/min; UV 245 nm) shows the presence of 796 mg of product (yield 91%).

Example 5. Doramectin (compound of formula (I))

A solution of 500 mg of the compounds of the decree the minutes are added dropwise 2.1 ml of 2 M solution of KOH in methanol. The reaction mixture, stirring well, then maintained at -35oC for 1 hour to complete the reaction, then add 252 mg glacial acetic acid, dissolved in 0.5 ml of methanol. Stop cooling and for 1 hour and added dropwise 2.2 ml of water to initiate crystallization. After stirring at room temperature for 1 hour, the crystals are filtered, washed with two portions of 0.5 ml of a mixture methanol:water 7:3 and dried in a vacuum oven. After recrystallization from aqueous methanol receive 272 mg 93,8-th product (output 70,6%). The retention time of 6.5 min (HPLC, Ultrasphere ODS 5, 25 cm x 4.6 mm (Beckman); methanol:acetonitrile:water 860:51:89; 1.5 ml/min; UV 245 nm).

Example 6. Doramectin (compound of formula (I))

To a solution of 6.6 g of compound indicated in the heading of example 3, in 75 ml of methanol, cooled to -15oC, add 75 ml of a saturated solution of ammonia in methanol. The mixture was kept at -15oC for 5.5 hours to complete the reaction, then rinsed with nitrogen for 30 minutes. Volatile impurities are removed in vacuo, the obtained yellowish oil was dissolved in 45 ml of methanol and the solution is purified by filtration. To the solution are added dropwise 5 ml of water to initiate crystallization. The mixture is after further stirring for 1.5 hours the solid is filtered off and washed with two portions of 2.5 ml of a mixture methanol:water (7:3). After drying in a vacuum oven at room temperature for 24 hours to obtain 3.75 g 87,7% of product (yield 64%).

Example 7. The compound of formula (III), where R1is acetyl and R2is hydrogen

To a solution of 20 g of compound of formula (II) in 200 ml of methylene chloride by passing nitrogen was added 28.6 g of triethylamine and 1.06 g of 4-dimethylaminopyridine. The reaction mixture is cooled to -5oC...0oC and added dropwise within 5 minutes add 14.5 g of acetic anhydride. The mixture was incubated for 20 minutes at 0oC to complete the reaction, then quenched at 0oC 200 ml saturated sodium bicarbonate solution. The organic layer re-extracted twice with saturated sodium bicarbonate solution and dried (MgSO4). The solvent is removed under vacuum; gain of 22.2 g of 95% of the product as a white solid (yield 96,7%). (HPLC, Ultrasphere ODS 5, 25 cm x 4.6 mm (Beckman); methanol:acetonitrile:water 902:36:62; 1.0 ml/min; UV 245 nm).

Example 8. The compound of formula (III), where R1is acetyl and R2is hydrogen

To a suspension of 5 g of compound of formula (II) in 100 ml of toluene by passing nitrogen gain of 7.24 g of triethylamine and 266 mg of 4-dimethylaminopyridine. The mixture is cooled to -15oC and added dropwise within 5 minutes and, quenched with 50 ml saturated sodium bicarbonate solution and stirred for 1 hour at room temperature. The organic layer re-extracted with twice 50 ml of saturated sodium bicarbonate solution and dried partial distillation of the solvent, using a nozzle Dean-stark. The solution is analyzed by HPLC (Ultrasphere ODS 5, 25 cm x 4.6 mm (Beckman); methanol:acetonitrile: water 902:36:62; 1.0 ml/min; UV 245 nm), it contains at 5.27 g of product (yield of 96.5%).

Example 9. The compound of formula (III), where R1is acetyl and R2is hydrogen

To a solution of 20 g of compound indicated in the heading of example 7 in 200 ml of ethyl acetate by passing nitrogen was added 60 ml of anhydrous pyridine, then add 22,2 g O-p-tailgatenation, then roll a yellow precipitate. The mixture is heated to the boiling temperature and incubated for 2 hours with good stirring to complete the reaction and cooled to room temperature. Volatile impurities are removed in vacuo, the residue is partitioned between 150 ml of ethyl acetate and 150 ml of water, the organic layer is extracted twice with saturated sodium bicarbonate solution and once with water. The ethyl acetate layer dried (MgSO4), the solvent is removed under vacuum. The residue is dissolved in 25 ml ethyl acetate and 5 ml of methyl the two silica gel cartridges. Eluent composition has hexane:ethyl acetate:methylene chloride 70: 25: 5. The fractions containing the product are pooled, the solvent is removed in vacuum; obtain 19 g of the product as a yellowish solid (yield of 82.6%). The retention time of 17.1 minutes. (HPLC, Ultrasphere ODS 5, 25 cm x 4.6 mm (Beckman); methanol: acetonitrile:water 902:36:62; 1.0 ml/min; UV 245 nm).

Example 10. 5,4"-O-diacetyl-doramectin (compound of formula (IV), where R1is acetyl)

A suspension of 9.5 g of calcium carbonate is heated to 156o-158oC in continuous transmitting current of nitrogen. Then to this mixture of 18.7 g of compound indicated in the heading of example 9; heat, stirring and passing nitrogen continued for 25 hours. After cooling to 50oC the mixture is filtered and the solvent is removed in vacuum. The residue is dissolved in 50 ml of methylene chloride, add 25 ml of ethyl acetate and 70 ml of hexanol, the solution is filtered and chromatographic using system Prep-500 (Water Associates) equipped with two silica gel cartridges. Eluent composition has hexane:ethyl acetate:methylene chloride 70:25:5. The fractions containing the product are pooled, the solvent is removed in vacuum; gain of 13.7 g 87% of the product as a yellow solid (yield 74.8 percent). The retention time of 11.1 minutes. (HPLC, Ultrasphere ODS 5 is ktin (compound of formula (IV), where R1is acetyl)

To a solution of 504 mg of the compound indicated in the heading of example 9, in 25 ml of 2-ethoxyethyl ether add 250 mg of calcium carbonate. The mixture is heated with good stirring and continuous transmitting current of nitrogen to 183o-185oC. After keeping at 183o-185oC for 1.5 hours analysis of the mixture shows the presence of 364,4 mg (yield 84.7 per cent).

Example 12. Doramectin (compound of formula (I))

To a solution of 7.3 g of compound indicated in the heading of example 10, in 145 ml of anhydrous tetrahydrofuran by passing nitrogen is cooled to -72oC, add 59,5 ml of 0.1 M solution of triethylborohydride lithium in tetrahydrofuran. The reaction mixture was stirred at -72oC...-70oC for 1 hour and slowly warmed to room temperature to complete the reaction. The mixture is then quenched with 75 ml of water and 75 ml of methylene chloride and stirred for 1 hour. The layers separated, the aqueous layer was extracted with 75 ml of methylene chloride; the combined organic layers are washed with saturated sodium bicarbonate solution and twice with 75 ml. of the Solvent is removed in vacuum, the residue is dissolved in 54 ml of methanol and the solution is purified by filtration. To the solution is added dropwise within 20 minutes was added 6.9 ml of water initiation cu the drops are added 6.2 ml of water. The mixture is stirred for 18 hours, the solid phase is separated by filtration, washed in a vacuum furnace, get 4,27 g 89,5% of product (yield of 57.3%).

Example 13. The influence of the bandwidth of nitrogen to the reaction heat removal

To a solution of 5.3 g of compound indicated in the heading of example 2, in 55 ml of 2-methoxyethanol ether add 2.5 g of calcium carbonate. The mixture is heated to 157oC with good stirring and continuous transmitting current of nitrogen through the gas distribution tube. After keeping at 154o-157oC for 30 hours the mixture is cooled to room temperature, filtered and analyzed by HPLC Ultrasphere ODS 5, 25 cm x 4.6 mm (Beckman); methanol: acetonitrile:water 559:383:58; 1.5 ml/min; UV 245 nm). The analysis shows the presence 3,93 g 5,4"-O-Diphenoxylate-doramectin (yield of 84.3%).

This experiment was repeated under the same conditions, including equal loading of the connection specified in the header of example 2, but without passing nitrogen. The yield in this case was 47.5%, showing a positive effect of the bandwidth of nitrogen.

Biological activity

The use of compounds as anti-helminth funds demonstrated the activity of these compounds on the model in v is ocent cleaning from the nematode Trichostrongylus colubriformis in rats infected with these parasites.

Anthelminthic efficacy against T. colubriformis in rats after oral doses of 1 mg/kg

Connection - Cleanup from worms compared to the result achieved from the drug intake

Example 7 - 100

Example 9 - 75

Example 10 - a

1. Derivatives doramectin General formula III

< / BR>
where R1- (C2-C5-alkanoyl or phenoxyacetyl, optionally substituted by 1-3 (C1-C4)-alkyl, (C1-C4)-alkoxy or halogen;

R2is hydrogen or phenoxythiocarbonyl, optionally substituted by 1-3 (C1-C4)-alkyl, (C1-C4)-alkoxy or halogen.

2. Connection on p. 1, where R1- (C2-C5) alkanoyl, phenoxyacetyl or (C1-C4-alkylphenoxy and R2is hydrogen or (C1-C4-alkylenediamines.

3. Connection on p. 1, where R1- C6H5OCH2C (=O).

4. Connection on p. 3, where R2- hydrogen.

5. Connection on p. 3, where R2- p-CH3-C6H5OC (=S)-.

6. Connection on p. 2, where R1- CH3C (=O)-.

7. Connection on p. 6, where R2- hydrogen.

8. Connection on p. 6>/BR>where R1- (C2-C5-alkanoyl or phenoxyacetyl, optionally substituted one to three (C1-C4)-alkyl, (C1-C4)-alkoxy or halogen.

10. Connection on p. 9, where R1- (C2-C5-alkanoyl, phenoxyacetyl or (C1-C4-alkylphenoxy.

11. Connection on p. 10, where R1- CH3C (=O)-.

12. Connection on p. 10, where R1- C6H5OCH2C (=O)-.

13. The method of producing doramectin formula I

< / BR>
characterized in that it comprises the following successive stages:

(a) interactions of the compounds of formula II

< / BR>
with allermuir agent of the General formula (R4CO)2O or R4COX, where R4- phenoxymethyl, optionally substituted one to three (C1-C4)-alkyl, (C1-C4)-alkoxy or halogen and X Is Cl or Br, with the formation of compounds of General formula III

< / BR>
where R1- phenoxyacetyl, optionally substituted one to three (C1-C4)-alkyl, (C1-C4)-alkoxy or halogen;

R2is hydrogen;

b) interaction of the compounds of formula III, where R1defined above and R2- H, with a compound of formula R3OC (=S)X, where R WHICH X Is Cl or Br, with the formation of the compounds of formula III, where R1defined above and R2phenoxythiocarbonyl, optionally substituted one to three (C1-C4)-alkyl, (C1-C4)-alkoxy or halogen;

C) interaction of the compounds of formula III, where R1and R2defined above, in an inert solvent at a temperature of 150 - 200oC for 2 to 48 h with calcium carbonate when the continuous supply of inert gas with the formation of compounds of General formula

< / BR>
where R1defined above, and

g) the interaction of the compounds of formula IV, where R1defined above, with a base in alcohol with the formation of compounds of formula I.

14. The method according to p. 13, where the inert gas is nitrogen.

15. The method according to p. 14, where the base - NH3, KOH, KCN, Na2CO3, NaHCO3or NaOAC.

16. The method of producing doramectin formula I

< / BR>
characterized in that it comprises the following successive stages:

a) interaction of the compounds of formula II

< / BR>
with allermuir agent of the formula (R4CO)2O or R4COX, where R4defined above, X Is Cl or Br, with the formation of compounds of General formula III

< / BR>
where R1defined above;

R2- water>
-H, with a compound of formula R3OC (=S)X, where R3defined above and X Is Cl or Br, with the formation of the compounds of formula III, where R1- (C1-C4-alkanoyl and R2- phenoxythiocarbonyl, optionally substituted by 1-3 (C1-C4)-alkyl, (C1-C4)-alkoxy or halogen.

C) interaction of the compounds of formula III, where R1- (C1-C4-alkanoyl and R2defined above, in an inert solvent at a temperature of 150 - 200oC for 2 to 48 h with calcium carbonate when the continuous supply of inert gas with the formation of compounds of General formula IV

< / BR>
where R1- (C1-C4-alkanoyl;

g) the interaction of the compounds of formula IV, where R1- (C1-C4-alkanoyl, with lithium aluminum hydride, cyanoborohydride sodium or triethylborohydride lithium in an inert solvent with the formation of compounds of formula I.

17. The method according to p. 16, where the inert gas is nitrogen.

18. The method according to p. 17, characterized in that it includes the interaction of the compounds of formula IV, where R1- (C1-C4-alkanoyl, triethylborohydride lithium.

19. The method according to p. 18, characterized in that it comprises mixing a specified seacure (-78) - (-70)oC, the shutter speed for a given temperature from -78oC to -70oC for from 15 minutes to 1 hour and the subsequent increase in temperature to room.

20. The method of obtaining the derived doramectin General formula IV

< / BR>
where R1defined in paragraph 1, characterized in that it includes the interaction of the compounds of formula III

< / BR>
where R1defined above and R2- phenoxythiocarbonyl, optionally substituted one to three (C1-C4)-alkyl, (C1-C4)-alkoxy or halogen in an inert solvent at a temperature of 150 - 200oC for approximately 2 h with calcium carbonate when the continuous supply of inert gas.

21. The method according to p. 20, where the inert gas is nitrogen.

22. A method of treating mammals suffering from parasitic diseases, by introducing derived secondary metabolites of microorganisms Vita Streptomyces avermitilis, characterized in that the injected antiparasitic effective amount of the compounds under item 1.

23. A method of treating mammals suffering from parasitic diseases, by introducing derived secondary metabolites of microorganisms Vita Strepromyces avermitilis, characterized in that the injected antiparasitic effective if

 

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