LT3676B - Macrolide compounds - Google Patents

Abstract

Compounds are described

Description

The present invention relates to novel compounds, antibiotics, processes for their preparation and pharmaceutical compositions based thereon.

In our patent specification no. 216646 / United Kingdom / We describe the antibiotic S541 which can be distinguished from the novel Streptomyces sp. fermentation products.

We have found another class of antibiotic activity compounds that can be obtained by chemically modifying antibiotic S541.

In one aspect, the invention relates to compounds represented by the formula:

OR ³ and their salts, wherein R ¹ represents a methyl, ethyl or isopropyl group; R represents a hydrogen atom, an alkyl group or Cį.g C ₃ _ ₈ alkenyl group; OR ³ represents a hydroxyl group or a substituted hydroxyl group having up to 25 carbon atoms; and = NOR is in E - configuration.

The term alkyl or alkenyl, as applicable to a group or moiety thereof in the compounds of Formula I, means that this group is linear or branched.

When compounds of formula I R 1 is C ₁ _ ₈ alkyl group, it may be eg. a methyl, ethyl, n-propyl, i-propyl or t-butyl group, most preferably a methyl group.

When R is a C ₃ _ ₈ alkenyl group which may be e. allyl group.

When the OR group in the compounds of formula 1 is a substituted hydroxyl group, it may be an acyloxy group [i.e., the group -OCOR ⁴ , -OCO ² R ⁴ , -OCSOR ⁴ (wherein R ⁴ is an aliphatic, arylaliphatic or aromatic group, e.g. alkyl, alkenyl, alkynyl, cycloalkyl, arylalkyl or aryl group) or formyloxy group, -OR ⁵ group (where R ⁵ is the same as mentioned for R ⁴ group), OSO ² R ⁶ group (where R ⁶ is C 2-4 alkyl or C 6-10 aryl group), cyclic or acyclic acetaloxy group , An OCO (CH 2) n CO ² R ⁷ group (where R ⁷ is a hydrogen atom or group as in R ⁴ and n can be zero, 1 or 2) or an OCONR ⁸ R ⁹ group (where R ⁸ and R each independently can be a hydrogen atom or C4_ ₄ alkyl group, for example. methyl).

When R ⁴ or R ⁵ are alkyl groups they may be e.g. alkyl groups, e.g. methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl or n-heptyl and may be substituted. When R ⁴ is a substituted alkyl group, it may be substituted e.g. one or more halogen (eg. chlorine or bromine atoms), or carboxyl, Ο ₄ _ ₄ alkoxy (eg methoxy, ethoxy), phenoxy or sililoksilo group. When R ⁵ is a substituted alkyl group, it may be substituted by cycloalkyl, e.g. cyclopropyl, a group.

5

When R or R are alkenyl or alkynyl groups, they may be e.g. C ₂ _ ₈ alkenyl as an allyl group, or a C ₂ _ ₈ alkynyl groups.

When R ⁴ or R ⁵ are cycloalkyl groups they may be e.g. C ₃ -C ₁₂ cycloalkyl groups as C ₃ . ₇ cycloalkyl, e.g. cyclopentyl group.

When R ⁴ or R ⁵ are arylalkyl groups they have from 1 to 6 carbon atoms in the alkyl residue and the aryl groups can be carbocyclic or heterocyclic and have from 4 to 15 carbon atoms, for example phenyl. Examples of such groups include phen-C 1-4 alkyl, e.g. benzyl groups.

When R ⁴ or R ⁵ are aryl groups they may be carbocyclic or heterocyclic and have 4 to 15 carbon atoms, e.g. may be a phenyl group.

When R ⁴ is substituted with a silyloxy group, the silyl group can include three groups, the same or different, from alkyl, alkenyl, alkoxyl, cycloalkyl, arylalkyl, aryl and aryloxy groups. These groups may be as defined above for R ⁴ , particularly suitable for methyl, t-butyl and phenyl groups. Typical examples of such silyloxy groups are trimethylsilyloxy and t-butyldimethylsilyloxy.

Where -OR ³ is a group -OSO ₂ R ⁶ , it may, e.g. be methylsulfonyloxy or para-toluenesulfonyloxy.

When -OR ³ is a cyclic acetaloxy group it may have from 5 to 7 atoms in the ring and e.g. may be tetrahydropyranyloxy.

When -OR ³ is a group represented by the formula OCO (CH 2) nCO ² R ⁷ , it may be e.g. OCOCO2R OCOCH2CH2CO2R ^7a or ^7a-type group, wherein R ^7a is hydrogen or C] _4 alkyl (eg. Methyl or ethyl) group.

The salts of the compounds of formula 1 which have an acidic group are bases such as alkali metal salts such as sodium and potassium.

In the compounds represented by formula 1, the group R ¹ should be an isopropyl group.

The group OR ³ should be methoxy-carbonyloxy or acetoxy, methoxy, oxy. In general, the compounds of formula 1, when OR is oxy, are most suitable.

Preferred compounds of formula 1 in the present invention are compounds wherein R is isopropyl, R is methyl and OR ³ is hydroxy, acetoxy or methoxycarbonyloxy.

As noted above, the compounds of the invention may be used as antibiotics. The compounds of the invention may also be used as intermediates for the preparation of other products. In this case, the -OR ³ group must be a protected hydroxyl group and the invention particularly encompasses such protected compounds. It should be emphasized that said group should have few side functionalities and no additional reaction centers and allow selective recovery of the hydroxyl group. Examples of such protected groups are well known and described e.g. in Protective Groups in Organic Synthesis by Theodora W.Greene, (WileyInterenee, New York 1981) and Protective Groups in Organic Chemistry by JFW McOmie (Plenum Press, London, 1973). Examples of protected hydroxyl groups in OR ³ are phenoxyacetoxy, silyloxyacetoxy (e.g. trimethylsilyloxyacetyloxy and t-butyldimethylsilyloxyacetoxy), and silyloxy groups such as trimethylsilyloxy and t-butyldimethylsilyloxy. The present invention relates to compounds having to groups 3676B which are mainly used as intermediates. Other groups, such as acetoxy groups, may be used as protecting hydroxyl groups and may also be present in the final active compounds.

The compounds of the invention have an antibiotic activity, e.g., an anthelmintic, e.g. against nematodes, and in particular anti-endoparasitic and anti -ectoparasitic activity.

Endoparasites and ectoparasites infect humans and many animals and infect farm animals such as pigs, sheep, rhinoceroses, as well as birds (such as chickens and turkeys), horses, rabbits and domestic animals such as. dogs, cats, guinea pigs, hamsters and so on. Parasitic infection in living organisms causes anemia and weight loss, leading to considerable economic losses.

Examples of endoparasites infecting said animals and / or humans include Ancylostoma, Ascaridia, Ascaris, Aspicularis, Brugia, Bunostomum, Capillaria, Chabertia, Cooperia, Cyathostomes, Dictyocaulus, Dracunculus, Enterobius,

Heteraki,

Dirofilaria,

Haemonchus,

Gastrophilus,

Loa, MetaHyostrongylus,

Nematodirus, Nematospiroides, Nippostrongylus, Oesophagostomum, Onchocerca, Ostertagia, Oxuris, Parafilaria, Parascaris, Probstmyria, strongylus, Necator,

Strongylus,

Toxascaris, Trichinelia, Wuchereria.

Strongyloides, Syphacia, Thelazia, Toxocara, Trichenoma, Trichostrongylus, Trichuris, Triodontophorus, Uncinaria and

Exemplary ectoparasites that infect animals and / or humans include arthropod parasites such as stinging insects, flies, fleas, blood sucking insects, mites and other double-winged insects.

Such ectoparasites may be e.g. Ambylomma, Anopheles, Boophilus, Chorioptes, Culepipiens, Culliphore, Demodex, Dmalinia, Dermatobia, Haemothobia, Haemotopinus, Haemophysalia, Hyaloma, Hypoderma, Ixodes, Linognathus, Lucilia, Melophagus, Oestrus, Pobius, Otobius Solenopotes, Stomoxys and Tabanus.

The compounds of the invention have been found to be effective against many endo- and ectoparasites both in vitro and in living organisms. The antibiotic activity of the compounds of the invention may be demonstrated against the nematodes Caenorhabditis elegans and Nematospiroides pelvis.

An important compound of the invention is represented by Formula 1 wherein:

R ¹ is a methyl group, R ² is a methyl group and OR ³ is a methoxy group.

Another important compound of the invention is represented by Formula 1 wherein:

R is an ethyl group, R is a methyl group and OR is a hydroxyl group.

A very important compound of the invention is represented by Formula 1 wherein:

R is an isopropyl group, R is a methyl group and OR is a hydroxyl group.

A compound of the formula wherein R ¹ is ethyl, R ² is methyl and OR ³ is hydroxyl is active against a large proportion of endo- and ectoparasites. Yes, for example.

this compound has been found to be active in vivo against parasitic nematodes such as Ascaris, Cooperia curticei, Cooperia oncopora, Cyathostomes, Dictyocaulus viviparus, Dirofilaria immitis, Gastrophilus, Haemanchus contortus, Nematodirus battus, Nematodirus helviatinus, Nematodirus spp. Oestophaostomum, Onchocera gutturosa, Ostertagia circumcincta, Ostertagia ostertagi, Oxyuris egui, Parascaris eguorusm, Probstmyria, Edentatus, Strongylus vulgaris, Toxocara canis, Trichostrongylus vitinus, Trichostrongylus axei, Triodontai , Anopheles stevensi, Boophilus dicolarartus, Boophilus microplus, Chorioptes ovis, Culexpipiens molestus, Damalinia bovis, Dermatobia, Haemotopinus, 'Hypoderma, Linognathus vituli, Lucilia sericata, Psoroptes ovis, Rhipicephalus app.

The compounds of the invention may also be used as insecticides, acaricides and nematocides in agriculture, gardens, public places and warehouses. Pests living on the soil or on agricultural crops (cereals, cotton, tobacco, vegetables, fruits, roots) can be eradicated by the compounds of the invention. Examples of such pests include Aphis Myzus persicae, Aulacorthum circumflexum,

Nephotettix cinciticeps, Nilparvata lugens, Panonychus ulmi, Phorodon humuli, Phyllocoptruta oleivora, Tetranychus urticea and the family Trialeuroides; nematodes from the families Aphelencoides, Globodera, Heterodera, Meloidogyne, and Panagrellus; butterflies - Heliothis, Plutella and Spodoptera; long-tailed Anthonomus grandis and Sitophilus granarius; beetle Tribolium castaneum; flies Musca domestica; ants; termites; Pear psylla; Thrips tabaci; cockroaches like Blatella germanica and Periplaneta americana and mosquitoes like Aedes aegypti.

It has been found that a compound of formula 1 wherein R ¹ is isopropyl group, R ² is methyl group and OR ³ is hydroxyl group is active against Tetranychus urticae (bean dwelling), Myzus persicae (Chinese cabbage leaf), Heliothis virescencis (ant. cotton leaves), Nilparvata lugens (rice paddy), Musca domestica (sugar and cotton in boxes), Blatella germanica (in food boxes), Spodopter exiqua (on cotton leaves) and Meloidogyne incognita.

The compounds of the invention may also be used as fungicides, e.g. against Candida sp. strains, e.g. Candida albicana and Candida glabrata and against yeast like Saccharomyces carlsbergensis.

The present invention provides compounds of Formula 1 as above which may be used as antibiotics. Partially, these compounds can be used in the treatment of animals and humans infected with endo- and ectoparasites, ectoparasitic and fungal infections, as well as in agriculture, forestry, plantation maintenance as pesticides for pest control and as acaricides and nematocides. These compounds can generally be used as pesticides in other cases, e.g. in warehouses, buildings and other public places where pests are present. In general, these compounds can be used both in the organism in which they live and in the areas where they live and against the pests themselves.

The compounds of the invention may be prepared by a variety of routes for their further use in veterinary or medical applications, and thus the invention also relates to pharmaceutical compositions comprising compounds of the present invention suitable for veterinary and medical use. Such compositions may be formulated in conventional manner using one or more suitable carriers or diluents. The compositions of the invention encompass such forms as are particularly suitable for internal administration of drugs, including administration to the mammary gland, as well as compositions for oral, rectal, topical, intramuscular, implantable, ophthalmic, nasal, genito-urinary administration. organism.

The compounds of the invention may be administered by medical or veterinary administration via injection, which may be in single-dose, ampoule or other container for single-dose or in multi-dose containers. If necessary, preservatives may be included in such compositions. Formulations for injection may be formulated as suspensions, emulsions, or solutions in water or other carrier vehicle, and may contain agents to stabilize the compositions, e.g. suspending, stabilizing, emulsifying and / or dispersing agents. Alternatively, the active ingredient may be in the form of a sterile powder for constitution with a suitable vehicle, e.g. sterile, pyrogen-free water, and such a preparation procedure is performed prior to use.

Fatty carriers include polyatomic alcohols and their esters, such as glycerol esters, fatty acids, vegetable oils, e.g. arachis oil, cottonseed oil or fractionated coconut oil, mineral oil fat, e.g. liquid paraffin, isopropyl myristate and ethyl oleate as well as other similar compounds. Other carriers such as formal glycerol, propylene glycol, polyethylene glycols, ethanol and glycofurol may also be used. Common nonionic, cationic, and anionic surfactants may be used alone or in combination in preparing the composition.

Formulations for veterinary use may also be formulated as injections into the mammary glands (to maintain a sustained, slow, sustained release of the active component), and may be formulated to provide rapid release of the active component. These compositions may be sterile solutions or suspensions in water or fat, optionally with a thickening or suspending agent, e.g. soft or hard paraffin, wax, 12-oxystearin, hydrogenated castor oil, aluminum stearate or glyceryl monostearate. Common nonionic, cationic, and anionic surfactants may be used alone or in combination in preparing the composition.

The compounds of the invention intended for medical or veterinary use may be presented in forms suitable for oral use, i. in the form of solutions, syrups, emulsions or suspensions, or as a dry powder, followed by mixing with water or another suitable vehicle, if necessary with a coloring or flavoring agent. Other solid compositions such as tablets, capsules, cakes, beads, pastes, powders, granules or pre-mixed ready-to-use formulations may also be used. Solid and liquid compositions for oral use may be prepared by conventional techniques well known to those skilled in the art. Such compositions may include one or more pharmacologically acceptable carriers or excipients in solid or liquid form. Examples of pharmacologically acceptable carriers for use in solid forms include binding agents (such as gelatinized starch, polyvinylpyrrolidone or hydroxypropylmethylcellulose), excipients, and

(such as lactose, microcrystalline cellulose or calcium phosphate), lubricants (such as magnesium stearate, talc or silica), disintegrants (such as potato starch or sodium starch glycolate) or wetting agents (such as sodium lauryl). The tablets may be coated by techniques well known and practiced in the art.

Examples of pharmacologically acceptable additives for use in liquid form include suspending agents (e.g., sorbitol syrup, methylcellulose or hydrogenated edible fats), emulsifying agents (e.g., lecithin or acacia), non-aqueous vehicles (e.g., almond oil, fatty esters, or ethyl alcohol). ) and preservatives (e.g., methyl or propyl paraoxybenzoates or sorbic acid), and stabilizing agents may be included in such prepared forms.

Oral pastes may be prepared by known techniques. Examples of pharmacologically acceptable additives in paste formulations include suspending and gelling agents, e.g. aluminum distearate or hydrogenated castor oil; dispersing agents, e.g. polysorbates; non-aqueous vehicles, e.g. arachis oil, fatty esters, glycols or macroglycols; stabilizing and solubilizing agents. The compounds of the present invention can be used in veterinary medicine by mixing them in 1 solid or liquid food ration.

The compounds of the invention may be administered orally for veterinary purposes in the form of liquid drugs, suspensions, solutions or dispersions of the active ingredient, together with a pharmacologically acceptable carrier or excipient.

The compounds of the invention may also be embodied in the form of suppositories, i.e. in dosages suitable for the manufacture of candles for veterinary or medical use, or in the form of pessaries, i.e. activating materials used in the production of common pessaries.

lubricating and / or gel lubricants are produced

The compounds of the invention may be adapted for topical use in veterinary and medical applications, e.g. in the form of ointments, creams, lotions, shampoos, powders, sprays, drops (eg nose and eye drops). Ointments and creams may be formulated with an aqueous or non-aqueous vehicle with the addition of suitable disintegrating agents. For eyes using sterile technology using sterile components. Formulations for surface wetting may be formulated for veterinary purposes using organic solvents or in aqueous suspension, and may include agents to accelerate the dermal adsorption process, and those compositions may include agents to provide solubilization, stabilization, or preservation and in one way or another to improve storage. and / or ease of use.

Lotions may be formulated with an aqueous or non-fatty base and will generally include one or more emulsifying, stabilizing, dispersing, thickening or coloring agents.

The powder may be prepared in any powdered carrier. The drops may be prepared using an aqueous or anhydrous carrier and may also include one or more dispersing, stabilizing, solubilizing, or suspending agents. They can also turn on a preservative.

The compounds of the present invention, when administered by inhalation, may be formulated as an aerosol.

The total daily dose of the compounds of the invention used in medicine and veterinary medicine should be in the range of from 1 to 2000 kg / kg body weight, preferably from 5 to 800 kg / kg, and may be divided into individual doses, e.g. 1 to 4 times a day. It should be noted that the dosage depends on the age and condition of the patient, the organism to be treated, the route of administration and the form used. The dosages required for a given organism may be calculated using conventional methods, e.g. comparing the activity of the given compound with a known antibiotic.

The compounds of the invention may be used in any conventional manner in agriculture or gardens, and in such manner. the present invention encompasses all those compositions containing the compounds of the invention which are useful in agriculture and in the treatment of gardens and plantations. Such ready-to-use forms activate anhydrous or liquid compositions, e.g. dusts, powders; granules; liquid compositions; emulsions; watering solutions; seed soaking solutions; fatty concentrates; fat solutions; injections; spraying, spraying and fogging compositions.

In general, such compositions will activate the active compound in association with a suitable carrier and diluent. Such carriers may be liquid or solid and may be used either for dispersing the compound or for preparing a dispersing composition for use in a liquid formulation. Such preparation methods are well known to those skilled in the art and can be obtained e.g. mixing and / or comminuting the active ingredient or ingredients with a carrier or diluent, as a solid carrier, diluent or surfactant.

Suitable solid carriers for e.g. duets, granules or powders may contain substances from natural mineral fillers, e.g. diatomite, talc, kaolinite, montmorillonite, profililite or atapulgite. If desired, the dispersed silicic acid or the dispersed adsorbent polymers may be incorporated into the composition. Granular carriers with adsorptive properties may be in the form of porous material (e.g. pumice, crushed brick, sepiolite or bentonite) and non-porous material (eg calcite or sand). Preformed granular formulations that can be used and may be of organic or inorganic origin (dolomite and crushed plant residues) are suitable.

Suitable solvents as carriers or diluents include aromatic carbohydrates, aliphatic carbohydrates, alcohols and glycols or their ethers, esters, ketones, acid amides, polar solvents, optionally unoxidized vegetable oils and water.

Common nonionic, cationic, or anionic surfactants, e.g. ethoxylated alkylphenols and alcohols, salts of alkali metals and sulfonates of alkylbenzenesulfonic acids, lignosulfonic acids or polymeric phenols having good emulsifying, dispersing and / or wetting properties, may be used either in their pure form or in said compositions.

Optionally, the compositions may include stabilizers, anti-adhesive agents, viscosity regulators, fillers and adhesives, photostabilizers, as well as fertilizers, food stabilizers and other active substances. The compounds of the present invention may also be used in combination with other insecticides, acaricides and nematocides.

In the above formulas, the active ingredient concentration will generally be in the range 0.01% to 99% by weight, preferably between 0.01% and 40% by weight.

Industrial products are generally manufactured in the form of concentrated compositions which need to be diluted to 0.001% to 0.0001% by weight prior to use.

The amount of compound used depends on factors such as the type of pest and the level of infestation. In general, suitable application rates are from 10 g / ha to 10 kg / ha, preferably from 10 g / ha to 100 kg / ha for killing mites and insects and from 50 g / ha to 10 kg / ha for killing nematodes.

The compounds of the invention may be used with other active ingredients. If necessary, the compounds of the invention may be used in combination with other known anthelmintic preparations. By combining the formulations of the invention with other anthelmintic agents, a greater number of parasitic diseases can be combated. In this way, the possibility of combating parasitic infections for which the products used alone are ineffective or ineffective can be realized.

The compounds of the invention may be prepared by the methods described below. In some of these methods, it is necessary to protect the hydroxyl group at position 5 of the starting material before carrying out the reaction described. In such cases, it is subsequently necessary to remove this protection in order to obtain the compound of the invention as the final product. For this purpose, conventional methods of applying and removing guards which, e.g. described in Grino and Mak-Ami's book e.

According to one aspect of the invention there is provided a process for the preparation of compounds of Formula I (A), which comprises reacting a compound of Formula 2:

(2) with a compound represented by Formula H ₂ NOR or a salt thereof (wherein R ^{2 is} as defined above) and optionally further deprotecting a compound represented by Formula 1 when OR ³ represents a protected hydroxyl group, and if necessary, obtaining a salt of this compound.

The oxidation reaction may be carried out in an aqueous or anhydrous medium, usually at a temperature in the range of -20 ° to + 100 ° C, e.g. -10 ° to + 50 ° C. It is preferable to use the salt of the reagent represented by the formula H ₂ NOR ² . Where salt is used, the reaction may be carried out in the presence of an acid-binding agent.

The solvents used are water and water miscible solvents such as alcohol (e.g. methanol, ethanol), amides (e.g. N, N-dimethylformamide, N, N-dimethylacetamide or hexamethylphosphoramide), ethers (e.g. cyclic ethers such as tetrahydrofuran or dioxane, and acyclic ethers such as dimethoxyethane or diethyl ether), nitriles (e.g. acetonitrile), sultans (e.g. sulfolane), hydrocarbons such as hydrocarbon halides (e.g. methylene chloride) and esters such as ethyl acetate, as well as mixtures, consisting of two or more such solvents.

Where the reaction is carried out in an aqueous medium, the appropriate acid, base or buffer may be used to buffer the medium at pH 2-9.

Suitable acids for this purpose include inorganic acids such as hydrochloric or sulfuric acid, and carboxylic acids such as acetic acid. Examples of suitable bases include alkali metal carbonates and bicarbonates such as sodium bicarbonate; hydroxides as sodium hydroxide and alkali metal carboxylates as sodium acetate. A suitable buffer is a solution consisting of sodium acetate and acetic acid.

The compounds represented by Formula 2 are either known compounds disclosed in British Pat. 2176182, or may be derived from known compounds described herein using standard procedures.

According to another aspect of the present invention there is provided another process (B) for the preparation of compounds of Formula 1 wherein R ² represents a C 1-6 alkyl group or C ₃ . _The alkenyl group ₈ and OR ³ represent a substituted hydroxyl group, and the essence of this process is the reaction between a compound represented by the formula wherein OR ³ represents a hydroxyl group and a reagent whose function is to replace the hydroxyl group, if necessary, to form a salt.

Acylation, formylation, sulfonation, etherification, silylation, and acetal formation reactions can be carried out using standard methods as described below.

Thus, for example, the acylation reaction may be carried out using an acylating agent, e.g. an acid represented by the formula R ⁴ COOH or an active derivative thereof, such as an acid halide anhydride (e.g. acid chloride); anhydride or active ester or an active derivative of a carboxylic acid of formula R OCOOH or a thiocarboxylic acid of formula R ⁴ OCSOH.

Acylation reactions using acid halide anhydrides or anhydrides may allow conditions where the acid-binding agent, e.g. such as tertiary amine (e.g., triethylamine, dimethylamine or pyridine), inorganic bases (e.g., calcium carbonate or calcium bicarbonate), and oxines, such as lower 1,2-alkylene oxides (e.g., ethylene oxide or propylene oxide), which bind hydrogen halides formed as a result of the acylation reaction.

The acylation reaction using acids must generally be carried out in the presence of a condensing agent, e.g. carbodiimide as N, N'-dicyclohexylcarbodiimide or Netyl-N 'γ-dimethylaminopropylcarbodiimide; a carbonyl compound such as carbonyldiimidazole; or isoxazole salts such as N-ethyl-5-phenylisoxazole perchlorate.

The activated ester can be obtained in situ using, e.g. 1-hydroxybenzotriazole in the presence of conLT 3676 B densifying agent, as noted above. Alternatively, the activated ester may be obtained in advance.

The acylation reaction can be carried out in an aqueous or anhydrous medium at temperatures ranging from -20 ° C to + 100 ° C, e.g. -10 ° to + 50 ° C.

The formylation reaction can be carried out using an active formic acid derivative, e.g. N-formylimidazole or mixed anhydride of formic acid and acetic acid under standard reaction conditions.

The sulfonation reaction can be carried out using an active sulfuric acid derivative represented by the formula R ⁶ SO ³ H, e.g. sulfonyl halide, e.g. chloride R ⁶ SO ³ Cl, or sulfo anhydride. The sulfonation reaction is usually carried out in the presence of an acid-binding agent in the reaction medium as previously described. The etherification reaction may be carried out using a reagent of formula R ⁵ Y (wherein R ⁵ is as previously described and Y represents a leaving group such as chlorine, bromine or iodine, or a hydrocarbylsulfonyloxy group such as methylsulfonyloxy or thiazyloxy; dichloroacetoxy group). This reaction may be carried out using Grignard's reagent as methylmagnesium halide, e.g. methylmagnesium iodide, or using trialkylsilylmethylmagnesium halide, e.g. trialkylsilylmethylmagnesium chloride, thereby obtaining magnesium alkoxide, followed by treatment with a reagent of formula R ⁵ Y and subsequent reaction.

Alternatively, the reaction may be carried out using a silver salt such as silver oxide, silver perchlorate, silver carbonate or silver salicylate, and mixtures of these salts, and this system may be particularly useful where the etherification process is carried out using an alkyl halide, e.g. methyl iodide.

The etherification reaction may be carried out in a solvent such as an ether, e.g. diethyl ether.

Acetal formation can be accomplished using a cyclic or acyclic vinyl ether. This method is particularly suitable for the preparation of tetrahydropyranyl ethers using a dihydropyran reagent or alkyl ethynyl ether as a reagent for 1-ethoxyalkyl ethers. This reaction must be carried out in the presence of a strong acidic catalyst, e.g. inorganic acid such as sulfuric acid or in the presence of an organic sulfuric acid such as para-toluenesulfonic acid in a non-hydroxylic, water-free solvent.

The solvents used in the above reactions may include: ketones (e.g. acetone), amides (e.g. N, N-dimethylformamide, N, N-dimethylacetamide or hexamethylphosphoramide), ethers (e.g. cyclic ethers such as tetrahydrofuran or dioxane), and acyclic ethers such as dimethoxyethane or diethyl ether), nitriles (e.g. acetonitrile), hydrocarbons such as hydrocarbon halides (e.g. methylene chloride) and esters such as ethyl acetate, and mixtures of two or more such solvents.

The silylation reaction can be carried out using a silyl halide (e.g. chloride), preferably in the presence of a base in the medium, as imidazole triethylamine or pyridine, using a solvent such as dimethylformamide.

The carbamoylation reaction to give compounds of formula 1, wherein OR ³ represents a group represented by the formula OCONR ⁸ R ⁹ , may be carried out using a suitable acylating (i.e., carbamoylating) agent. Suitable bamoylating agents of karLT 3676 B which may be used to prepare compounds wherein one of R and R is a hydrogen atom and the other is an alkyl group are isocyanates of the formula R ¹⁰ NCO (wherein R ¹⁰ is an alkyl group). The carbamoylation reaction is carried out in the presence of a solvent or mixture of solvents selected from the group consisting of: hydrocarbons (e.g. aromatic hydrocarbons such as benzene or toluene), halogenated hydrocarbons (e.g. dichloromethane), amides (e.g. formamide or dimethylformamide), such as ethyl acetate), ethers (e.g. cyclic ethers such as tetrahydrofuran or dioxane), ketones (e.g. · acetone), sulfoxides (e.g. dimethylsulfoxide), or mixtures of these solvents. This reaction can be carried out at temperatures ranging from -80 ° C to the reflux temperature of the reaction medium e.g. 100 ° C, preferably in the range -20 ° to + 30 ° C.

The presence of a base such as a tertiary organic base, e.g. tri (lower alkyl) amine (e.g., triethylamine).

Another suitable carbamoylating agent is cyanic acid, which may be prepared in situ, e.g. from an alkali metal cyanate as sodium cyanate and the presence of an acid, e.g. strong organic acids like trifluoroacetic acid. Cyanic acid effectively corresponds to the isocyanate compounds mentioned above wherein R ¹⁰ is hydrogen and thereby helps to obtain from their compounds of formula 1 their carbamoyloxy analogs (i.e. compounds represented by formula 1 wherein OR ³ is an OCONH ₂ group).

Alternatively, carbamoylation may be accomplished by reaction with phosgene or carbonyldiimidazole followed by reaction with ammonia or a suitable substituted amine in aqueous or anhydrous reaction medium.

compounds of formula in which OR ³ is a group of the formula OCO (CH 2) n CO 2 R ^7, can then be realized seilinant corresponding 5-hydroxy compound with acid of the formula HO 2 C (CH _{2) n} CO ₂ R ^7, or its reactive derivative to a procedure, described above.

According to another aspect of the invention is the compounds of formula 2 wherein R is a ₄ Ο _ ₃ alkyl group or

C ₃ -C ₈ alkenyl group, a process for the preparation of a compound of formula 1 wherein R is a hydrogen atom and OR is a substituted 2 2 hydroxyl group and an etherifying agent R Y (wherein R is C ₁ -C _{8). 8} alkyl or C ₃ _ ₈ alkenyl group, and Y are as described above); optionally deprotecting a compound of formula 1 wherein OR ³ is a protected hydroxyl group, and further providing a salt thereof.

The etherification reaction may be carried out using magnesium alkoxide using Grignard reagent as methylmagnesium halide and subsequent treatment with R Y reagent. Alternatively, the reaction may be carried out in the presence of a silver salt such as silver oxide, silver perchlorate, silver carbonate or silver salicylate and a mixture of these salts, or in the form of a base such as potassium carbonate or sodium hydride. The etherification reaction may be carried out in an organic solvent, such as an ether, e.g. diethyl ether, tetrahydrofuran or dioxane, or amide, e.g. dimethylformamide, hexamethylphosphoric triamide, or in a mixture of these solvents at ambient temperature. Under these conditions, the oxime group configuration is virtually unchanged after the etherification reaction.

According to another aspect of the present invention there is provided another process (D) for the preparation of compounds of formula 1 wherein OR ³ is a hydroxyl group and a compound of formula (3) having the essence of a compound

reduction with subsequent salt formation, if necessary.

The reduction reaction can be accomplished using a reducing agent which can stereoselectively reduce the 5-keto group. Suitable reducing agents include borohydrides, such as alkali metal borohydrides (e.g., sodium borohydride) and lithium alkoxyaluminium hydrides (e.g., lithium tributoxyaluminium hydride).

The reaction using a borohydride reducing agent is carried out in the presence of a solvent such as an alkanol, e.g. isopropyl alcohol or isobutyl alcohol, typically at -30 ° to + 80 ° C, e.g. At 0 ° C. The reaction using lithium alkoxyaluminum hydride is carried out in the presence of a solvent such as ether, e.g. tetrahydrofuran or dioxane, at -78 ° to 0 ° C.

The intermediates represented by Formula 3 may be obtained from the 5,23-diketone represented by Formula 4

previously) in an amount equivalent by applying the oxidation reaction conditions described above for the preparation of compounds of formula 1.

compounds of formula I may be obtained by oxidation of a compound represented by formula 5

This reaction may be carried out using an oxidizing agent for the modification of the secondary hydroxyl group to give the compound of formula 1.

Suitable oxidizing agents include: quinones in aqueous media, e.g. 2,3-dichloro-5,6-dicyano-1,4-benzoquinone or 2,3,5,6-tetrachloro-1,4-benzoquinone; oxidizing agents that activate chromium (6-valence), e.g. sodium or pyridine dichromate or chromium trioxide in pyridine medium, preferably in the presence of a phase transfer catalyst; oxidizing agent activating 4-valent manganese, e.g. manganese dioxide in dichloromethane; N-halosuccinimide, e.g. N-chlorosuccinimide or N-bromosuccinimide; dialkylsulfoxide in the presence of an oiling agent such as N, N'-dicyclohexylcarbodiimide, halide, e.g. oxalyl chloride; or a pyridine-sulfur trioxide complex.

The reaction may be carried out in a suitable solvent, which may be selected from the group consisting of: ketones, e.g. acetone; ethers, e.g. diethyl ether, dioxane or tetrahydrofuran; hydrocarbons, e.g. hexane; halogenated hydrocarbons, e.g. chloroform or methylene chloride; or esters, e.g. ethyl acetate or substituted amide, e.g. dimethylformamide. A combination of these solvents, both in pure form and in water, can be used. The choice of solvent depends on the oxidizing agent used in the reaction.

This reaction can be carried out in a temperature range of -80 ° to + 50 ° C.

The compounds of formula I may be obtained, for example, by culturing Streptomyces thermoarchaensis culture NCIB 12015 (deposited on 10 September 1984 at the National Industrial and Marine Bacteria (UK, Aberdeen, Tori Research Station) or a mutant thereof) and isolating the compound from the fermentation broth thus obtained.

The organism Sterptomyces thermoarchaensis can be grown by conventional methods, e.g. in the presence of appropriate sources of carbon, nitrogen and mineral for assimilation in the medium. Suitable sources of carbon, nitrogen, and minerals for assimilation can be in the form of simple or complex nutrients, e.g. as described in British Patent No. 2166436. Suitable media for incorporating said materials are described in Preparation No. 1 below.

Cultivation of Streptomyces is generally carried out at temperatures between 20 and 50 ° C, preferably between 25 and 40 ° C, by aeration and agitation, e.g. shaking and stirring. The medium may initially be inoculated with a small amount of the sporulated suspension of microorganisms, but to avoid growth retardation, the vegetative inoculum may be prepared by inoculating a small amount of culture with the sporulated organism, and the resulting vegetative inoculum is then introduced into the enzymatic medium or, more preferably, one or more stages of inoculation in which growth occurs before transfer to the main fermentation medium. In general, the fermentation is carried out at pH 2.5 to 8.5.

The fermentation can be carried out for 2 to 10 days, e.g. 5 days.

The compounds of formula I may be isolated from the enzymatic broth thus obtained by conventional methods of separation and isolation. Various fractionation methods can be used e.g. adsorption - elution, precipitation, fractional crystallization and solvent extraction, which can be combined in various ways. Solvent extraction and chromatography have been found to be most suitable for the isolation of the compounds. A process for preparing compounds of Formula 5 employing the following procedures is provided in Preparation no. 1, described below.

According to another aspect of the invention there is provided a process for the preparation of compounds of formula (E) wherein OR ³ is a hydroxyl group, which comprises deprotecting a compound of formula 1 wherein OR ³ is a protected hydroxyl group, as described above.

Yes, for example. an acyl group such as an acetyl group may be removed by an alkaline hydrolysis reaction, e.g. using sodium or potassium hydroxide in an alcoholic aqueous solution; or by acid hydrolysis, e.g. using concentrated sulfuric acid in methanol medium. Acetyl groups such as tetrahydropyranyl may be removed by acid hydrolysis (e.g., acetic or trifluoroacetic acid or dilute inorganic acid). The thread groups may be removed using fluoride ions (e.g., tetralkylammonium fluoride, as fluoride), hydrogen fluoride, in acetonitrile solution, or acidic, such as paratoluene sulfuric acid (e.g., methanolic medium). The arylmethyl groups may be removed by treatment with Lewis acid (e.g. boron trifluoride etherate) in the presence of a thiol (e.g. ethanolic) in a suitable solvent such as dichloromethane at room temperature.

tetra-n-butylammonium in aqueous

The acid salts represented by Formula 1 can be obtained by conventional methods, e.g. by treating an acid with a base or converting one salt into another by an ion-exchange reaction.

The present invention is illustrated but not limited to the Preparations and Examples below, wherein temperatures are in ° C, 'L' represents liter and EtOH means ethanol.

In the Preparations and Examples below, the compounds are provided as derivatives of known factors A, B, C and D. Factor A is a compound described in ForLT 3676 B mule 6, wherein R ¹ is isopropyl, R ³ is hydrogen; Factor B is a compound represented by Formula 6 wherein R ¹ is methyl, R ³ is methyl; Factor C is a compound represented by Formula 6 wherein R is methyl, R is vanyl; Factor D is a compound represented by Formula 6 wherein R ¹ is ethyl, R ³ is hydrogen.

1-5-Keto-Factor A Preparation

Spores of Streptomyces thermoarchaensis NCIB 12015 were inoculated into agar medium prepared from the following components:

g / L

Yeast extract (Oxoid L21) 0.5

Malt Extract (Oxoid L39) 30.0

Mycopeptone (Oxoid L40) 5.0

Agar No.3 (Oxoid L13) 15.0

Distilled water to 1 L pH ~ 5.4 and spores were incubated for 10 days at 28 ° C.

The mature culture was then covered with 6 ml of 10% glycerol solution and the spores and mycelium were sterilized with a sterile instrument. Aliquots of 3.4 ml of the resulting suspension were transferred to sterile polypropylene tubes, which were subsequently sealed and stored in liquid nitrogen vapor until needed.

Two 250 ml Erlenmeyer flasks containing 50 ml of inoculated medium were prepared as follows:

g / L

D-glucose 15.0

Glycerin 15.0

Soy peptone 15.0

NaCl 3.0

CaCO ₃ 1.0

Distilled water to 1 L (initially pH = 6.7; pH = 7.0 before autoclaving with aqueous sodium hydroxide. After autoclaving pH = 7.3)

Each flask was inoculated with 0.2 ml of spore suspension taken from test tubes.

The flasks were then incubated for 3 days at 28 ° C, rotating at 250 rpm with a 50 mm diameter radius.

The contents of both flasks were used to inoculate a 70 L fermenter containing 40 L of a similar medium with the addition of 0.06% by volume of polypropylene 2000. Polypropylene 2000 was required to control foaming. The fermentation process was carried out at 28 ° C with stirring and aeration, maintaining oxygen saturation above 30% · After 24 hours of fermentation, a 9 L portion of broth was loaded into a 700 L fermenter containing 450 L medium prepared as follows:

D-glucose

Maltese Dextrin (MD3OE) Arkasoy 50

Molasses

K ₂ HPO ₃

CaCO ₃

Silicone-525 (Dow Corning) g / L

2.8

27.8

13.9

1.7

0.14

1.39

0.06% (by volume)

The pH should be 6.5 before sterilization.

The fermentation was carried out at 28 ° C by aeration and stirring. Polypropylene 2000 was added to control foaming and the pH was lowered to 7.2 by addition of H ₂ SO ₄ until the culture matured. The fermentation took 5 days.

450 liters of broth were centrifuged (Westfalia KA 25 centrifuge) and the supernatant was replaced with 20 L of water. 25.5 kg of fiber were separated and mixed for 1 hour. using a sufficient amount of methanol in a Silversom BX mixer and resulting in 75 L of material. The resulting suspension was filtered and the solid phase was re-extracted with 35 L of methanol and then filtered. The total filtrate (87 L) was diluted with water (40 L) and extracted with petroleum ether (30 L) at 60-80 °. After 30 minutes the phases were separated in a Westfalia MEM 1256 centrifuge and the lower methanol phase was repeatedly extracted at 60-80 ° with petroleum ether (30 L) followed by addition of 40 L water. After separation, the lower phase was extracted with petroleum ether (30 L) at 60-80 ° C. The combined petroleum ether phases (85 L) were concentrated by passing three times through a Pfaudler evaporator 8.8-12v-27 (vapor pressure 0.1 bar, vapor temperature 20 °, vapor temperature 127 °). The concentrate (9 L) was dried using sodium sulfate followed by evaporation under reduced pressure at 40 ° C.

The resulting oil (130 g) was dissolved in chloroform to give 190 mL of product passed through a Merck 7734 column filled with silica 60 (200 x 4 cm). The column was washed with chloroform (500ml), then eluted with a mixture of chloroform and ethyl acetate (3: 1), after which approximately 40ml fractions were collected after the initial 1400ml passage.

Fractions 32-46 were mixed and evaporated to give an oil (21.2 g). Fractions 47-93 were mixed and evaporated to give an oil (20.1 g) which was dissolved in a mixture of chloroform and ethyl acetate (3: 1) to a total volume of 53 mL, which was then passed through a Merck 7734 column packed with silica. 60 (200 x 4 cm) suspended in a mixture of chloroform and ethyl acetate (3: 1), after which fractions of about 40 ml were collected after the initial 1400 ml was omitted. Fractions 22-36 were mixed and evaporated to give an oil (3.1 g), which was mixed with the oil from fractions 32-46 in the first column. The resulting mixture was dissolved in boiling methanol (4 mL) and then added to 20 mL of hot propan-2-ol followed by crystallization.

After crystallization, the solution was evaporated to give an oil which was dissolved in an equal volume of methylene chloride followed by loading onto a Merck Kieselgel 60 (sieve 70-230 ASTM, Art. No. 7734) column (30x2.2cm) loaded in methylene chloride. The layer was washed with two volumes of methylene chloride, then eluted with two volumes of a mixture of chloroform and ethyl acetate (1: 1). Evaporation of the eluent allowed to obtain an oil which was dissolved in methanol, followed by chromatography on a high performance liquid chromatograph Spherisoro S5 ODS-2 (250mm x 20mm, Phase Sep. Ltd.). 5ml samples were pumped into the column for 1 min and the column contents eluted with a mixture of acetonitrile and water (7: 3) as follows:

Time (min) Amount (ml / min)

0.00 0.00 } Enter 1.00 0.00 } start 1.10 30.00 39.90 30.00 40.00 35.00 75.00 35.00

The material eluted from the chromatographic column was analyzed by ultraviolet spectroscopy at 238 nm.

Evaporation of the pooled fractions with an elution peak of 33.4 min gave 33.4 mg of compound -1-5-ketofactor A as a solid.

E.I. molecular ion at 610 was detected by mass spectroscopy and showed characteristic fragments at: 592, 574, 556, 422, 259 and 241.

Example [E] -Methoxyimine Factor A

a) 5,23-diketo-factor A

An ice-cold solution obtained by mixing 1.2 ml of sulfuric acid with 120 mg of sodium dichromate in 2 ml of water was added over 15 minutes to an ice-cooled mixture of 200 mg of 5-keto-factor A and 15 mg of tetrabutylammonium in 4 ml of ethyl acetate. After one hour, the resulting mixture was diluted with ethyl acetate, after which the organic phase was washed with a saturated aqueous solution of sodium bicarbonate. The dried organic phase was evaporated and the resulting resin was purified by chromatography on Merck Keiselgel 60, sieve 230-400 (100 mL).

Elution with 10% ethyl acetate in dichloromethane gave 86 mg of the title compound as a yellow foam; δ (CDCl ₃ ) 6.57 (m, 1H);

250 (s, 2H); and 1.89 (m, 3H).

b) 5-keto, 23 [E] -methoxyimine Factor A

475 mg of 5,23-diketo-factor A, 69 mg of methoxylamine hydrochloride and 135 mg of sodium acetate were dissolved in methanol. After 1.5 hours at room temperature, the solution was stored for 16 hours at -18 ° C, diluted with ethyl acetate and rinsed thoroughly with 1N hydrochloric acid, water and brine. The dried organic phase was evaporated and the resulting yellow foam was chromatographed on Merck Keiselgel 60, 230-400 mesh (120 ml). Elution of the column with hexane / ethyl acetate 4: 1 gave 255 mg of the title compound as a yellow foam [α] p ^χ + 80 ° (c 1.2O, CHCl ₃ ), k _max (EtOH) 241nm (ε 27500), u _max (CHBr ₃ ), 3530, 3460 (OH) 1708 (C = O), 1676 (C = CC = O), 986 (CO), δ (CDCl ₃ ) 6.58 (s; 1H), 3.80 (s; 1H), 3.58 (m; 1H), 3.30 (d14; 1H), 1.00 (d6; 3H), 0.96 (d6; 3H), 0.92 (d6; 3H).

c) 23 [E] -methoxyimin-factor A

1) 6.5 mg of sodium borohydride was added to an ice-cooled solution of 83 mg of 5-keto, 23 [E] -methoxyimine factor A in 20 mg of isopropanol. The resulting yellow mixture was stirred in an ice bath for 35 minutes, then diluted with ethyl acetate and rinsed thoroughly with 1N hydrochloric acid, water and brine. The dried organic phase was evaporated and the resulting yellow gum was chromatographed on a Merck Keiselgel 60 sieve 230-400 (60 mL). Elution of the column with 2: 1 hexane: ethyl acetate gave 58 mg of the title compound as a yellow foam.

Crystallization from hexane gave the title compound with a melting point of 203 °, [α] _D ²¹ + 133 ° (c 1.12, CHCl ₃ ), X _raax (EtOH) 244nm (ε 26200), δ (CDCl ₃ ) 4.29 ( t7; 1H), 3.84 (s; 3H), 3.29 (d15, 1H).

2) 50 mg of 5-keto, 23 [E] -methoxyimino-factor A dissolved in Imi tetrahydrofuran was added to a cooled solution to -78 ° of 261 mg of lithium tris-t-butoxyaluminium hydride dissolved in 3 ml of tetrahydrofuran. After 0.75 h at -78 °, the resulting mixture was diluted with 30 mL of ethyl acetate and washed thoroughly with 0.5N hydrochloric acid and water. The dried organic phase was evaporated and the resulting intermediate was chromatographed on a Merck Keiselgel 60 sieve 230-400 (40 mL). Elution of the column with 25% ethyl acetate in hexane gave the title compound as a white foam [a] p ² ! + 128 ° (c 0.95, CHC1 ₃ ), δ (CDC1 ₃ ) turns on

4.29 (t7; 1H), 3.84 (s; 3H), 3.29 (d15, 1H).

An example

23 [E] -methoxyimino-factor A, 5-acetate

A solution of 2.8 g of sodium acetate dissolved in 15 ml of water was added to a solution of 23-keto-factor A, 5-acetate (3.13 g in Example 18 of UK Patent 2176182) dissolved in methanol, followed by the addition of 3.01 g of methoxyamine hydrochloride. The resulting mixture was stirred at 28 ° for 1.5 h, diluted with ethyl acetate and washed thoroughly with 0.5 N hydrochloric acid, water and brine. The dried organic phase was evaporated to near dryness and the resulting white foam was chromatographed on a Merck Keiselgel 60, 230-400 sieve (600 ml). Eluting the column with hexane and ethyl acetate, 4: 1 ratio as a result was obtained 14.2 mg of the title compound as colorless foam, [a] _D ²¹ + 128 ° (c 1:35, CHC1 ₃₎ ^ _ax (EtOH) 244 nm (c _max 27,250) , while _max (CHBr ₃ ), 3560, 3480 (OH) 1733 (acetate), 1715 (C = O), 995 (CO), δ (CDCl ₃ ), 5.5-5.6 (m: 2H), 3.84 (S: 3H), 3.29 (d 15; H), 2.16 (S: 3H).

example

23 [E] -hydroxyimin-factor A, 5-acetate

The reaction between 23-keto-factor A, 5-acetate and hydroxylamine hydrochloride was carried out as in Example 1 above. The intermediate was chromatographed on Merck Keiselgel 60, 230-400 mesh. Elution of the column with 4: 1 ethyl acetate: acetonitrile gave the title compound as a colorless foam, [α] _D + 132 ° (c 1.21, CHCl ₃ ), (EtOH) 244 nm (ε ^ 27800), and _max (CHBr). ₃ ), 3565,

3470 (OH) 1733 (acetate), 1712 (C = O), 993 (CO), δ (CDCl ₃ ) 8.12 (S: 1H), 5.5-5.6 (m: 2H), 3.42 (d 15: H),

2.16 (s 3H).

Example [E] -methoxyimin-factor A

1.88 g of the product obtained in Example 2 were dissolved in methanol and the resulting solution was cooled in an ice bath, followed by the addition of 5.5 ml of aq. It was then diluted with ethyl acetate and rinsed thoroughly with 0.5N hydrochloric acid, water and brine. The dried organic phase was evaporated and the resulting foam was chromatographed on a Merck Keiselgel 60 sieve 230-400 (400 mL). Elution of the column with 2: 1 hexane: ethyl acetate gave 1.429 mg of the title compound as a colorless foam.

Crystallization from hexane gave the title compound with a melting point of 203 °, [α] + 132 ° (c 1.21, CHCl ₃ ), k _max (EtOH) 244 nm (s _max 29200), u _max (CHBr ₃ ), 3540 ( OH), 1708 (C = O), 992 (CO), δ (CDCl ₃ )

4.29 (t7: 1H), 3.84 (s: 3H), 3.29 (d15: 1H).

Example [E] -hydroxyimin-factor A

Hydrolysis of the product of Example 3 using the procedure described in Example 3 gave the product which was chromatographed on Merck Keiselgel 60, 230-400 mesh (400ml). Elution of the column with hexane / ethyl acetate 1: 1 gave the title compound as a colorless foam, [α] _D ²¹ + 140 ° (c 1.24, CHCl ₃ ), k _max (EtOH) 244 nm (s _max)

26700), u _max (CHBR ₃ ), 3565, 3490 (OH), 1710 (C = O), 994 (CO), δ (CDCl ₃ ) 8.11 (s: 1H), 4.29 (t7: 1H), 3.41 ( d15 (1H).

example

23 [E] -ethoxyimin-factor A

To a solution of 140 mg of sodium acetate dissolved in 3 ml of water was added a solution of 1 23-keto-factor A (200 mg, example 23 in UK Patent No. 2176182) and 126 mg of ethoxyamine hydrochloride dissolved in 20 ml of methanol. After 2 hours storage at 20 °, the resulting mixture was diluted with 40 mL of ether and washed with water. The dried organic phase was evaporated, and the resulting white foam was chromatographed on Merck Keiselgel 60, 230-400 mesh (90 ml). Elution of the column with 2: 1 hexane: ethyl acetate gave 189 mg of the title compound as a colorless foam, [α] θ ²¹ + 125 ° (c 1.00, CHCl ₃ ), k _max (EtOH) 244 nm (E _max 28200), u _max (CHBr ₃ ), 3540.3480 (OH), 1705 (C = O),

990 (CO), δ (CDC1 ₃₎ 4.30 (t7: H), 4.10 (KV7: 2H), 3.31 (DL5: H), 1.24 (t7: 3H).

The compounds of Examples 7,8,9 were obtained analogously from 23-keto-factor A using the corresponding alkoxyamine.

example

23 [E] -alyloxyimino factor A [α] _D ²¹ + 124 ° (c 1.17, CHCl ₃ ), λ _max (EtOH) 244 nm (s _max

28400) and _max (CHBr ₃₎ 3550, 3490 (OH), 1708 (CO), 990 (CO), δ (CDC1 ₃₎ 5.98 (m, lH), 5.28 (ddl7,2; H), 5.15 (dd9 , 2; 1H), 4.3-4.7 (m; 2H), 4.29 (t7; 1H), 3.36 (d14, 1H), the compound was prepared using allyloxyamine hydrochloride.

example

23 [E] -isopropyloxyimino factor A [α] _D ²¹ + 116 ° (c 0.97, CHCl ₃ ), λ _max (EtOH) 244 nm (ε _max 25000), u _max (CHBr ₃ ), 3550, 3490 (OH ), 1708 (C = O),

992 (CO), δ (CDC1 ₃₎ 4.2-4.4 (m, 2H), 3.30 (DL4; H), 1.21 (d7; 3H), 1.20 (d7; 3H), title compound was prepared using izopropiloksiamino hydrochloride.

example

23 [E] -butoxyimino-factor A [α] _D ²¹ + 115 ° (c 1.10, CHCl ₃ ), λ _max (EtOH) 244 nm (s _max

31800), u _max (CHBr ₃ ), 3540, 3460 (OH), 1708 (C = O), 992 (CO), δ (CDCl ₃ ) 4.28 (t ₆ ; 1H), 4.03 (m; 2H), 3.96 ( d6; 1H), 3.31 (d14; 1H), 0.9-1.1 (m; 15H), the compound was obtained using n-butoxyamine hydrochloride.

example

23 [E] -methoxyimino-factor A, 5-acetate

1) A 3 molar solution of methylmagnesium dissolved in 0.16 mL of ether was added to a solution of 120 mg of the product of Example 3 dissolved in 5 mL of hexamethylphosphoric triamide under nitrogen. 0.09 ml of iodomethane was then added and after one hour the resulting solution was diluted with 30 ml of ethyl acetate and rinsed thoroughly with 2N hydrochloric acid and water. The dried organic phase was evaporated and the resulting resin was chromatographed on a Merck Keiselgel 60 sieve 230-400 (80 mL). Elution on a column with a 2: 1 mixture of hexane and ethyl acetate gave the title compound as a white foam, [α] _D ²¹ + 123 ° (c 1.25, CHCl ₃ ), λ _max (EtOH) 245 nm (s _max 28200). The nuclear-magnetic resonance spectrum was as described in Example 2.

2) 0.082 g of the product of Example 3 was dissolved in ml of diethyl ether containing 0.4 g of silver oxide (obtained by mixing silver nitrate with 2M sodium hydroxide solution).

The resulting mixture was stirred for 2 hours at room temperature, then filtered and evaporated to give the intermediate as a yellow gum. The resulting resin was chromatographed on Merck 5717 and eluted with a 25: 1 mixture of dichloromethane and acetone. The main fraction was extracted, evaporated, and 0.059 g of the title compound was obtained. The nuclear-magnetic resonance spectrum was as described in Example 2.

Example [E] -methoxyimin-factor A, 5-methylcarbamate

0.13 ml of 125 mg of methyl isocyanate and 2 drops of triethylamine were added to 1350 ml of a solution of 23 [E] -methoxyimine-factor A dissolved in 0.75 ml of dimethylformamide. The flask containing the resulting solution was sealed and heated at 80 ° for 5.5 hours with stirring. The mixture thus prepared was poured into 1 50 ml of water and the resulting mixture was filtered. The precipitate was washed with 150 ml of water and extracted with 75 ml of dichloromethane. The resulting extract was dried over magnesium sulfate and then concentrated to give a yellow foam which was chromatographed on a silica gel column under medium pressure (125 g Merck Kieselgel 60, 230-400 mesh). Elution of the column with a 1: 1 mixture of hexane and ethyl acetate gave 206 mg of the title compound.

of the title compound as a white foam [α] _D + 99 ° (c 0.55, CH ₂ Cl ₂ ), _{λ max} (EtOH) 244.4 nm (s _max 28710), u _max (CHBr ₃ ), 3530 (OH), 3455 ( NH), 1720 (ester), 1720 + 1510 (carbamate), and 993 cm @ ¹ (CO); δ (CDCl ₃ ), 1.78 (s, 3H), 2.86 (d, 5Hz, 3H), 3.29 (d, 14Hz, 1H), 3.83 (s, 3H), 4.80 (sq, 5Hz, 1H) and 5.50 (m) , 2H).

Example [E] -methoxyimine-factor A, 5-methyl carbonate

Methyl chloroformate (0.7 mL of 1M dichloromethane solution) was added to a solution of 150 mg of 23 [E] -methoxyimine-factor A dissolved in mL dichloromethane and 0.3 mL pyridine at 0 ° C. The resulting mixture was stirred for 20 min. at 0-3 °, followed by the addition of 70 ml of dichloromethane and washing with 50 ml of 2N hydrochloric acid and 50 ml of water. The organic phase was dried over magnesium sulfate, after which the solvent was removed to give a foam which was chromatographed on a silica gel column under medium pressure (40 g, Merck Kieselgel 60, 230-400 mesh). Elution of the column with 30: 1 dichloromethane / ethyl acetate gave 127 mg of the title compound.

j unginio white colored foam form [α] _D ²¹ + 14 5 ° (c = 0.41, CH ₂ C1 ₂ ), k _max (EtOH) 244.4 nm (ε 31210), υ _ΜΧ (CHBr ₃ ), 3460+ 3540 (OH), 1742 (carbonate), 1710

(ester) and 992 cm @ ¹ (CO); δ (CDCl ₃ ), 1.82 (s, 3H),

3.29 (d 14Hz, 1H), 3.82 (s, 3H), 3.83 (s, 3H), 5.2-5.4 (m; 3H), 5.56 (s, 1H).

Example [E] -methoxyimin-factor D, 5-acetate

A solution of 251 mg of 23-keto-factor D, 5acetate (Example 119 in UK Patent No. 2176182), 250 mg of sodium acetate and 250 mg of methoxyamine hydrochloride dissolved in 40 ml of methanol was stored for 24 hours at 20 °, followed by concentration. to 10 ml, then diluted with 50 ml of ethyl acetate and washed with 0.5 N hydrochloric acid and water. The dried organic phase was evaporated, and the resulting yellow foam was chromatographed on Merck Keiselgel 60, 230-400 mesh (120 ml). Elution of the column with hexane gave 144 mg of the title compound as a colorless foam, k _max (EtOH) 244 nm (ε 26, 400) (cm ^-1 ) 3500 (OH), 1732 (OAc), 1710 (C = O), δ (CDC1 _3), 5:54 (m, 2H), 4.92 (m, lH), 3.84 (s, 3H), '3:32 (m, 1H), 3.30 (DL4; H), 2.17 (s, 3H), 1.91 ( d14, 1H),

1.76 (s; 3H), 1.63 (s; 3H), 1.51 (s; 3H), 1.01 (t7; 3H),

0.99 (d6; 3H), 0.92 (d6; 3H).

Example E [E] -methoxyimine-factor 140 mg of the product obtained in Example D and 0.6 ml of 1N sodium hydroxide were dissolved in 8 ml of methanol and the resulting solution was stirred in an ice bath for 1.5 hours. The resulting solution was diluted with 30 mL of ethyl acetate and washed with 1N hydrochloric acid and water. The dried organic phase was evaporated and the resulting yellow foam was chromatographed on a Merck Keiselgel 60 sieve 230-400 (50 mL). Elution of the column with hexane / ethyl acetate 2: 1 gave 105 mg of the title compound as a whitish foam, [α] D ²¹ + 96 ° (c 1.38, CHCl ₃ ); k _max (EtOH)

244nm (ε 26700); u _raax (CHBr ₃₎ ^(cm-1) 3550, 3500 (OH), 1710 (C = O), δ (CDC1 ₃₎ 4.93 (m, 1H), 4.30 (t6; H), 3.95 (d6; H) , 3.84 (s; 3H), 3.30 (d14; 1H), 3.27 (m; 1H), 1.88 (s; 3H), 1.64 (s; 3H), 1.52 (s; 3H), 1.01 (t7; 3H), 1.00 (d6; 3H), 0.92 (d6; 3H).

example

23 [E] -methoxyimin-factor B

A solution of 1 g of 23-keto-factor B (Example 19 in UK Patent No. 2176182), 400 mg of sodium acetate and 400 mg of methoxyamine hydrochloride was stirred for 20 hours at 20 ° and then concentrated to a volume of 10 ml. dilute and rinse with water. The organic phase was washed with 0.5N hydrochloric acid, then the dried organic phase was evaporated and the product was chromatographed on Merck Keiselgel 60, 230400 mesh (200 ml). Elution of the column with a 1: 9 mixture of ethyl acetate and dichloromethane afforded 500 mg of the title compound as a white foam, [α] _D ²¹ + 128 ° (c 1.09, CHCl ₃ ); _λmax (EtOH) 244 nm (ε 30100); u _raax (CHBr ₃ ) (cm ^-1 ) 3540, 3460 (OH), 1708 (C = O), δ (CDCl ₃ ) 5.46 (kv6; 1H), 4.03 (d6; 1H), 3.97 (d5; 1H) , 3.83 (s, 3H), 3.50 (s; 3H), 3.32 (m; 1H), 3.29 (dl4; 1H), 1.82 (s; 3H), 1.68 (d6; 3H), 1.00 (d6; 3H), 0.92 (d6; 3H).

example

23 [E] -methoxyimin-factor C

0.54 g of sodium acetate and 0.58 g of methoxyamine hydrochloride were added to a solution of 23-keto-factor C (1.97 g of Example 12 in UK Patent No. 2176182) dissolved in 30 ml of methanol to give 5 ml of water, resulting in The mixture of 1 21 of the title compound, [α] θ, was stirred for 30 min at room temperature. To the resulting solution was added 30 mL of ethyl acetate and 30 mL of 0.5N hydrochloric acid, and the resulting aqueous layer was extracted with 15 mL of ethyl acetate. The combined organic layers were washed with 0.5M hydrochloric acid, 5% saturated aqueous sodium bicarbonate, and 10% saturated aqueous sodium chloride, and the mixture was concentrated in vacuo to give a yellow foam. The resulting product was purified by chromatography on a silica gel Merck 9385 column, initially with dichloromethane. The column was then eluted with dichloromethane containing up to 10% of ethyl in the name _n to trigger the following signals:

7Hz); 3.96 (d, 1H, 10Hz) (c 1.0, CH ₃ OH); 1 H NMR 4.95 (m, 1H); 4.29 (t, 1H, (d, IH, 7 Hz), 3.85 (s, 3H [= NOCH _3]); 3.66

1.51 (s, 3H); 1.42 (t, 1H, 12Hz); and (CHBR ₃ )

3620-3340 cm ^-1 (-OH), 1711 cm ^-1 (C = O).

The following examples relate to compositions prepared in accordance with the present invention. The term active ingredient used below relates to the compounds of the present invention and may be e.g. the compound shown in Example 4.

Multiple Dose Parenteral Injection Sample% w / v (s)

Active ingredient

Benzyl alcohol

Polysorbate 80

Formal glycerol Water for injections

2.0

1.0

10.0

50.0 to 100.0

Range

0.1-6.0% w / v

The active ingredient is dissolved in Polysorbate 80 and formal glycerol. Benzyl alcohol is then added and the water is made up to volume. By standardized methods, the product is sterilized by filtration or packaging.

by autoclaving and

PO e.g. a sterile example of that

Active ingredient

Benzyl alcohol

Glyceryl triacetate

Propylene glycol% w / v (w / v)

4.0

2.0

30.0 to 100.0

Range

0.1-7.5% w / v

The active ingredient is dissolved in benzyl alcohol and glyceryl triacetate. It is then made up to volume with propylene glycol by the usual methods, sterile packing.

Product sterilization is carried out e.g. sterile filtration followed by an example

Active ingredient

Ethanol

Non-ionic surfactant (eg Synperonic PE L44 *)

Propylene glycol

ICI brand% weight / volume (s / t)

2.0

36.0% v / v

10.0 to 100.0

Range

0.1-7.5% w / v

The active ingredient is dissolved in ethanol and the surfactant. Subsequently, propylene glycol is added to the required volume. Sterilization of the product is carried out by conventional methods, e.g. followed by sterile filtration followed by sterile packaging.

example

Active ingredient Non-ionic surfactant (eg Synperonic PE L44 *)

Benzyl alcohol

Miglyol 840 ** 16.0

Water for Injections * ICI Brand% Weight / Volume (s / t)

2.0 0

Range

1-3.0% w / v

2.0

1.0% v / v to 100.0 ** Dynamit Nobel brand

The active ingredient is dissolved in Miglyol 840. The surfactant and benzyl alcohol are dissolved in a larger proportion of the water. The emulsion is then prepared by adding the fat solution to the aqueous solution while homogenizing the mixture by standard methods. Then make up to volume and prepare and package under sterile conditions.

Aerosol

% weight / volume (s / t) Range Active ingredient 0.1 0.1-3.0% w / v Trichiorethane 29.9 Trichlorofluoromethane 35.0 Dichlorofluoromethane 35.0

The active ingredient is mixed with trichloroethane and the resulting mixture is placed in an aerosol container. The top of the cylinder is purged with gaseous propellant and the valve assembly pressed. It is then filled with liquid propellant through the valve to the required weight. Built-in pressure head and cap.

Pills

Method of production - wet granulation

The active ingredient is Magnesium Stearate

Corn starch

Starch sodium glycolate

Sodium laurilsulfate

Microcrystalline cellulose for tablet coating mg

250.0

4.5

22.5

9.0

4.5 to 450.0 mg by weight

Sufficient 10% starch paste is added to the active ingredient to give the required wet mass suitable for granulation. The pellets are made and dried, then sieved, the remaining ingredients are added and tablets are formed.

If necessary, the tablets may be film-coated using oxypropylmethylcellulose or other similar material. A plasticizer or suitable paint may be incorporated into the film forming solution.

Veterinary pills for small / pet animals

Method of production-dry granulation mg

Active ingredient 50.0

Magnesium stearate 7.5

Microcrystalline cellulose for coating tablets 75.0 mg

The active ingredient is mixed with magnesium stearate and microcrystalline cellulose. The mixture is further compacted into balls and passed through a granulator to form granules which are compressed into tablets. If necessary, the tablets may be film-coated as described above.

Veterinary injections into the mammary gland

Active ingredient

Polysorbate 60

White beeswax Arachis oil

Where s / s means mg / dose

150.0

3.0% s / s)

6.0% s / s to 3 s / s)

91.0% w / w

Range 0.05-1.0 g)

g) up to 3 or 15 g)

Arachis oil, white beeswax and polysorbate 60 are heated to 160 ° with stirring. The resulting mixture is maintained at this temperature for 2 hours and then cooled to room temperature with stirring. The active ingredient is then added under sterile conditions and dispersed using a high speed mixer. Grind using a colloidal grinder. Hereafter the product

bottled sterile syringes • Veterinary tablets discharge with the slow of the active ingredient % s / s Range Active ingredient 0.25-2 g Colloidal silicon dioxide 2.0 required amount Microcrystalline cellulose to 100.0 mg to full volume The active ingredient is mixed with colloidal silicon

by microcrystalline cellulose using any suitable technology to distribute the active ingredient over the entire volume of the substance. The following composition is introduced into a pharmacologic device providing slow release of the active ingredient, which allows 1) sustained release of the active ingredient, or 2) rapid release of the active ingredient.

Veterinary dosage for oral use

Active ingredient Polysorbate 85

Benzyl alcohol Propylene glycol Phosphate buffer pH

Water% s / t Range

0.35 0.01-2% s / t

5.0

3.0

30.0 for support 6.0-6.5 to 100.0

The active ingredient is dissolved in Polysorbate 85, benzyl alcohol and propylene glycol. Add water and adjust pH to 6.06.5, if necessary, using phosphate buffer. Make up to volume with water. The resulting product is placed in a container for oral administration of the drug.

Veterinary paste for oral use

Active ingredient

Saccharin sodium

Polysorbate 85

Aluminum distearate

Fractionated coconut oil% s / s Range

4.0 1-20% s / s

2.5

3.0

5.0 to 100.0

The aluminum distearate is dispersed on heating in fractionated coconut oil and Polysorbate 85. The resulting mixture is cooled to room temperature and the saccharin sodium is dispersed in a fat carrier followed by dispersing the active ingredient. The resulting product is packaged.

Veterinary granules for use with food% s / s

Active ingredient 2.5

Calcium sulphate (hydrated) to 100.0

Range

0.05-5% s / s

The active ingredient is mixed with calcium sulfate. The wet process is used to obtain the pellets and the pellets are subsequently dried. The resulting pellets are placed in a suitable container.

Veterinary composition for watering

Active ingredient

Dimethylsulfoxide

Methyl isobutyl ketone

Propylene Glycol (and Pigment)% s / t Range

2.0 0.1-30%

10.0

30.0 to 100.0

The active ingredient is dissolved in dimethyl sulfoxide and methyl isobutyl ketone. Subsequently, propylene glycol and pigment are added to the required volume. The resulting composition is placed in a container suitable for irrigation.

Emulsifiable concentrate

Active ingredient Anionic emulsifier (eg Phenylsulfonate CALX) Nonionic emulsifier (eg Synperonic NP13 *) Aromatic solvent (egSolvesso 100)

50.0 g 40.0 g

60.0g to 1.0L * ICI brand

The ingredients are mixed until completely dissolved. 15th

Granules

a)

Active ingredient

You say

50.0 g 40.0 g

Gypsum beads (sieve 20-60) (eg Agsorb 100 A)

b)

Active ingredient

Synperonic NP 13 *

Gypsum beads (sieve 20-60) (eg Agsorb 100 A) up to 1.0 kg

50.0 g

40.0g to 1.0kg * ICI trademark

All ingredients are dissolved in a volatile solvent, e.g. methylene chloride, followed by granulation in a blender. Dried to remove solvent.

The pesticidal activity of the compounds of the invention was determined by testing them on various pests in their habitats according to the following procedure:

- the product was in liquid form. The forms were obtained by dissolving the product in acetone. The solutions were then diluted with water containing 0.1% or 0.01% wetting agent until the product concentration reached the required concentration.

The test methodology used for most pests has enabled the maintenance of a certain number of pests in their normal environment either by treatment with the environment (residual test) or, in the case of Tetranychus urticae, Myzus persicae, Nilparvata lugens and Musca domestika, by pests and environment treatment (contact test). In the case of Meloidogyne, the solution was applied to soil containing tomato seedlings which were attacked by nematodes, after which the number of root tumors was compared with the control plants.

According to these procedures, the compound represented by Formula 1 wherein R ¹ is isopropyl, R ² is methyl and R ³ is hydrogen is effective at concentrations of 100 ppm or less.

Claims (15)

DEFINITION OF INVENTION Compounds of formula 1 NOR ^{2 in} which R ¹ is methyl, ethyl or isopropyl; R ² is hydrogen atom, C ^ g alkyl group or C ₃ _ ₈ aikenilo group and = NOR group in the E-configuration; OR ³ is a hydroxyl group or a substituted hydroxyl group having up to 25 carbon atoms and their salts. 2. Compounds according to claim 1, wherein OR is methoxycarbonyloxy, acetoxy, methoxy or hydroxy. 3. Compounds according to claim 1, wherein OR ³ is a hydroxyl group. Compounds according to any one of the preceding claims, characterized in that R ¹ is an isopropyl group. Compounds according to any one of the preceding claims, characterized in that R is a methyl group. 6. Compounds according to claim 1, characterized in that R is an isopropyl group, R is a methyl group and OR ³ is a hydroxy group, an acetoxy group or a methoxycarbonyloxy group. Compounds according to claim 1, characterized in that R ¹ is an isopropyl group, R ² is a methyl group and OR ³ is a hydroxyl group. Compounds according to claim 1, characterized in that R is a methyl group, R is a methyl group and OR ³ is a methoxy group; or R ¹ is ethyl, R ² is methyl and OR ³ is hydroxy 9. A composition for use in medicine, which comprises an effective amount of at least one compound according to claim 1 in combination with one or more carriers and / or excipients. 10. A composition for veterinary use comprising providing an effective amount of at least one compound of claim 1 together with one or more carriers and / or excipients. 11. A composition for controlling pests, which comprises activating an effective amount of at least one compound according to claim 1 together with one or more carriers and / or excipients. A composition according to any one of the preceding claims 9 to 11, wherein it comprises an effective amount of at least one compound according to claim 7 together with one or more carriers and / or excipients. 13. A method of killing pests in agriculture, horticulture or forestry, or in warehouses, buildings, or other public places or pests, where the plants or vegetables are exposed, or the pests are directly exposed to one or more compounds according to claim 1. . 14. The method of claim 13, wherein said pests are insects, mites and nematodes. 15. A process for the preparation of a compound according to claim 1 wherein: A) Reaction of a compound represented by formula 2 OR ³ 12.3 with a reagent of formula H ₂ NOR or a salt thereof (R, R and OR are as defined in claim 1, followed by deprotection, where appropriate, of a compound of formula 1 wherein OR ³ is a protected hydroxyl group; B) to give a compound characterized by formula 1 wherein R is _Ο χ _ ₈ alkyl or C ₃ _ ₈ alkenyl group, and OR is a substituted hydroxyl group by reaction with the corresponding compound of formula 1 in which OR ³ is a hydroxyl group and the reagent, capable of change a hydroxyl group; C) to give a compound characterized by formula 1 wherein R is Ο ₃ _ ₈ alkyl or C ₃ _ ₈ alkenyl group, and OR is a substituted hydroxyl group by reaction with the corresponding compound of formula 1 wherein R is a hydrogen atom and OR ³ is substituted by a hydroxyl eterifikuojančio group and the reagent R ² Y (wherein R ² is _{0> 8} alkyl or C ₃ _ ₈ alkenyl group, and Y is a leaving group) with the next, if necessary, deprotection of a compound of formula 1 in which oR ³ is protected hydroxy; D) The compound of formula 1 is obtained wherein OR ³ is a hydroxyl group; OR is a hydroxyl group by deprotection of the corresponding compound represented by formula 1, wherein OR ³ is a protected hydroxyl group, or F) the salt of the acid of the formula I obtained by reaction between said acid and a base or by conversion of one salt to another by an ion exchange reaction.