SI8712186A - Process for obtaining new macrolide compounds - Google Patents

Abstract

The invention relates to a process for the preparation of new ones macrolide compounds having an antibiotic activity of the formula (l) wherein R 1 represents methyl, ethyl or isopropyl group; R 2 represents a hydrogen atom, C 1-6 alkyl a group or a C3-8 alkenyl group and is a group = NOR2 in the E-configuration; OR3 is a hydroxyl group or substituted hydroxyl group having up to 25 atoms carbon, and pharmaceutically acceptable salts thereof comprising the reaction of compounds of formula (II) with the reagent H2NOR2 or with its salt (R1, R2 and OR3 are as defined above), and where appropriate for the preparation of compounds of formula (I), wherein R2 is Cis alkyl or C3-8 alkenyl group and OR3 substituted hydroxyl group, a reaction of the compounds of formula (I) is carried out, wherein OR3 is a hydroxyl group, with a reagent for converting the hydroxyl group into a substituted hydroxyl a group, or for the preparation of compounds of formula (I), wherein R 2 is a C 1-6 alkyl group and a C 3-6 alkenyl group, a reaction of the compounds of formula (I) is carried out, wherein R2 is an atom hydrogen or OR3 substituted hydroxyl group, s etherification agent R2Y (wherein R 2 is C 1-6 alkyl and C3-8 aikenyl group, Y is a leaving group), and then, if necessary, remove the protecting group from protected hydroxyl group OR3 in a compound of the formula (I) and the compounds of formula (I) are optionally converted to their salt.

Description

The invention relates to the production of new pharmaceutically useful organic compounds, and in particular relates to a process for the preparation of new macrolide compounds with antibiotic activity.

TECHNICAL_PROBLEM

The invention solves the problem of making new macrolide compounds, powerful antibiotics, synthetically.

SITUATION .. TECHNICAL

The UK Patent Application from the same Applicant No.2166436 describes the production of an Antibiotic S541 that can be isolated from the fermentation product of a new Streptomyces Sp.

2? I§_? SOLUTIONS_TECHNICAL_PROBLEMS_SA_ EXAMPLES

The process of the invention provides novel macrolide, antibiotic compounds by chemical modification of the Antibiotic S541, compounds of formula (I):

or salts thereof, wherein R ¹ represents methyl, ethyl or isopropyl group 2

R represents a hydrogen atom, a C ^ g alkyl group or ^ ₃ _θ alkenyl smaller group; OR1 is a hydroxyl group or a substituted hydroxyl group having up to 25 carbon atoms; and group = NOR "is in E configurations and.

The term " alkyl " or " alkenyl " as a group or part of G ^ nfbe formula (I) indicates that the group is straight or branched.

-v · :.

in meals ·

- * ri. T * · '·

When R in the compounds of formula (I) represents a C 1-10 alkyl group. this may be, for example, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl or t-butyl, and preferably a methyl group.

When R ^{c is a} -3_g alkenyl group, it may be, for example, an allyl group.

When the OR group in the compounds of formula (I) is a substituted hydroxyl group, it may represent an aryloxy group / e.g.

A group of the formula -OCOR, -OCC ^ R or -OCSOR (wherein R is an aliphatic, araliphatic or aromatic group, for example, alkyl, alkenyl, alkyl.cycloalkyl, aralkyl or aryl group) /, formyloxy group, group - OR ³

4 6 (where R is as defined above for R), group -OSO_R (where

R is alkyl or 0θ_ ^ θ aryl group), a cyclic or acyclic acetalox group, a group 0C0 (CH ₂ ) _n CO ₂ R ^ (where R? Is a hydrogen atom or a group as defined for R ⁴ above and is zero, 1 or 2) or 8 9 8 9 an OCONR R group (wherein R and R can each independently represent a hydrogen atom or a C 1-6 alkyl group, e.g., methyl).

5

When R or R are alkyl groups, they may be, for example, _θ alkyl groups, e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl or n- heptyl wherein these alkyl groups are also substituted. When R is a substituted alkyl group it may be substituted with, for example, one or more, e.g., two or three halogen atoms (e.g., chlorine or bromine atoms), or with carboxy, alkoxy (n. or., methoxy, ethoxy), phenoxy or siloxy gruo. When R is a substituted alkyl group, it may be substituted with a cycloalkyl, e.g., a cyclopropyl group.

5

Kada su R or R is alkenyl or alkynyl groups, one may be prepared to be, for example, C ₂ _g alkenyl, e.g. allyl, or _C2 -8 ^alkynyl group.

5

When R or R are cycloalkyl groups, they may be, for example, ^C 3-12 cycloalkyl, such as cycloalkyl, e.g. cyclopentyl groups.

5

When R or R are aralkyl groups, they preferably have 1 to 6 carbon atoms in the alkyl group and aryl groups may be carbocyclic or heterocyclic and preferably contain 4-15 carbon atoms, e.g., phenyl Examples of such groups include ίεηθ ^ _θ alkyl e.g., benzyl / groups.

5

When R or R are aryl groups, they may be carboxylic or heterocyclic and preferably have 4-15 carbon atoms, and may be, for example, a phenyl group.

When R contains a silyloxy substituent, the silyl group may carry three groups which are the same or different and are selected from alkyl, alkenyl, alkoxy, cycloalkyl, aralkyl, aryl and aryloxy groups. Such groups may be 4 as defined above, and particularly include methyl, t-butyl and phenyl groups. Some examples of such silyloxy groups are • trimethylsilyloxy and t-butyldimethylsilyloxy.

When the -OR ^ group is -OSO ₂ R ^, these measures are, for example, methylsulfonyloxy or p-toluenesulfonyloxy.

When -OR ^ represents a cyclic acetaloxy group, it may be, for example, a group having 5-7 ring members and may be, for example, a tetrahydropyranyloxy group.

7

When OR represents the group 0C0 (CH ₂ ) CO ₂ R, it may be, for example, the group OCOCO ₂ R ^{/ a} or OCOCH ₂ CO ₂ R ' ^and where R ^ ^a represents a hydrogen atom or alkyl (e.g., methyl or ethyl) group.

Salts which may be formed with compounds of formula (I) containing an acidic group include salts with bases, e.g., alkali metal salts such as sodium and potassium salts.

In the compounds of formula (I), the group R1 is preferably an isopropyl group.

The OR group is preferably a methoxycarbonyloxy or, in particular, an acetoxy, methoxy or hydroxy group. Generally, compounds of formula (I) 3 in which OR is a hydroxy group are particularly preferred.

Important compounds of the invention are those of formula (I) wherein r is *

3 isopropyl group, R is methyl group and OR is hydroxy, acetoxy or methoxycarbonyloxy group.

As indicated earlier, the compounds of the invention can be used as antibiotics. The compounds of the invention can also be used as intermediates for making other active compounds. When the compounds of the invention are to be used as intermediates,

The -OR group may be a protected hydroxyl group and the invention is defined;

includes such protected compounds. It will be appreciated that such a group should have minimal additional functionality to avoid further reaction sites and should be such that a hydroxyl group can be selectively regenerated from it. . Examples of protected hydroxyl crump are well known and are described, for example, in Protective Groups in

Organic Synthesis by Theodore W Greene. (Wiley-Inuerscience, Nev.)

York 1981) and Protective Groups in Organic Chemistry cd J.F.W.

McOmis (Plenum Press, London, 1973). Examples of OR protected hydroxy groups include phenoxyacetoxy, silyloxyacetoxy, (e.g. trimethylsilyloxyacetoxy and t-butyldimethylsilyloxyacetoxy) and silyloxy groups such as trimethylsilyloxy and t-butyloxy. Compounds of the invention containing such groups will primarily be used as intermediates. Other groups, such as acetoxy, may serve as protected hydroxyl groups, but may also be present in the final active compounds.

The compounds of the invention have antibiotic activity, e.g., anthelmintic activity, for example, against nematodes, and in particular anti-endoparietal and anti-ectoparasite activity.

Ectoparasites and endoparasites infect humans and various animals, and are particularly prevalent in domestic animals such as pigs, sheep, goats and poultry (eg, chickens and turkeys), horses, rabbits, fun birds, caged birds, and the like domestic animals such as dogs, cats, guinea pigs, gerbils and hamsters. Parasitic livestock infection, which leads to anemia, poor nutrition and weight loss, is a major cause of economic loss worldwide.

Examples of genera andoparasites infecting such animals and / or humans are An c y los torna, Ascaridia, Ascaris, A.spicularis, Brngia ,, Bunostomum, Capillaria, Chabertia, Cooperia, Cyathostomes, Dictvocaulus, Ditrof ilaria.,. Dracunculus, Enterobius, Gastrophilus, Haemonchus, Heterakis, Hvostrongylus, Loa, Metastrongvlus, Necator, Nematodirus, Nematospiroiaes, Nippostrongylus, Oesophagostomum, Onhocerca, Ostertacia. Oxvuris, Parafilaria, Parascaris, Probstmavria, Strongvlus ,.

Strongvloides, Syphacia, Thelazia, Toxascaris, Toxocara, Trichonema,

Trichostrongvlus, Trichinella, Trichurus, Triodontophorus, Uncinaria and Wuchereria.

Examples of ectoparasites infecting animals and / or humans are arthropod ectoparasites such as biting insects, various flies, flies, mites, blood sucking insects, gundi, and other ciptero damage.

Examples of the genera of such ectoparasites that infect animals and / or humans are Ambvloma, Anopheles, Boophilus, Chorioptes, Culszpipiens, Culliphore, Demodex, Damalinia, Dermatobia, Haematobia, Haematopir.us, Hvaloma, Hvpoderma, Ophosus, Linophagus, Iathodes. Otobius, Otodectes, Psorergates, Psoroptes, Psoroptes, Rhipicephalus, Sarcoptes, Solenopotes, Stomoxys, and Tabanus.

The compounds of the invention have been found to be effective both in vitro and in you against the entire interval of endoparasites and ectoparasites. The antibiotic activity of the compounds of the invention can, for example, be demonstrated by their activity against loose living nematodes, e.g., Caenorhabditis elegans and Nematospiroides dubius.

An important active compound of the invention is that of formula (I) in which:

2 3

R is methyl, R is methyl and OR is methoxy.

Another important active compound of the invention is that of formula (I) in which j

3

R is ethyl, R is methyl and OR is hydroxyl.

A particularly important active compound of the invention is that of formula (I) in which:

2 3

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

2

A compound of formula (I) wherein R is an isopropyl group, R is a methyl group and OR1 is a hydroxyl group is active against a wide range of endoparasites and ectoparasites. For example, this compound has been found to be active in vivo against such parasitic nematodes such as Ascaris, Cooperia curticei, Cooperia oncophora, Cvathostomes, Dictvocaulus viviDarus, Dirofiliaria immitis, Gastroohilus, Haemonchus contortus, Nematodirus battus, Nematodiirorus helvetodi, helveti dubium; Nippostrongvlus braziliensis; Oesophaostomum; Triodontophorus and Uncinar stenic grafts of iparasitic beetles, mites, worms and yours such as Ambllgona hebraeum, Anopheles stevensi, Boophilus dicolarartus, Boophilus microplus, Choriopr.es dependent, Culexpipiens molestus, Dama liri a bocvusus, Dama liri a bocvusus, sericata, Psoroptes dependent, Rhipicephalus appendiculatus and Sarcopr.es.

The compounds of the invention are also used to combat insects, acarides and nematodes in agriculture, horticulture, forestry, public health and in stock products. It is useful to treat the damage caused by land and plant crops, including cereals (e.g. wheat, barley, corn and rice), cotton, tobacco, vegetables (e.g. soybeans), fruits (e.g. apples). , grapevines and lemons) as well as root crops (e.g., turnips, potatoes). Some examples of such crops are water mites and arides such as Aphis fabae, Aulacorthum circumflexum,

Mvzus persicae, Nephotettix cincticeps, Nilparvata lugens, Panonychus ulmi, Phorodon humuli, Phyllocoptruta oleivora, Tetranvchus urticae and and members of the genus Trialeuroides; nematodes such as members of the genus Aphelencoides, Globodera, Heterodera, Meloidogvna and Panagrellus; lepidoptera such as Heliothis, Plutella and Spodoptera; cereal grains; such as Anthonomus grandis and Sitophilus granarius; flour bugs such as Tribolium castaneum; flies such as Musca domesti ca; ants; leaf rodents; Pear psvlla; Thrips tabaci; cockroaches such as BlateIla cermanica and Periplaneta americana and mosquitoes such as Aedes aegvoti.

Certainly, we have found that the compound of formula (I) wherein R is

3 methyl group, R is methyl group and OR is hydroxyl group active against Tetranvchus urticae (supported on French bean leaf), Mvzus persicae (supported on Chinese cabbage leaf), Heliothis vire · scens (supported on cotton leaf), Nilaparvata lugens (supported on a rice plant), Musca domestica (in a plastic bowl with cotton wool / sugar solution), Blatella germanica (in a plastic pot with food granules), Spodoptera exigua (supported on a cotton leaf) and

Meloidogvne incognita.

The compounds of the invention can also be used as an anti-fungal agent, for example, against Candida sp. such as Candida albicans and Candida glabrata and against such yeast as Saccharo myces carlbergensis.

According to the invention, we provide compounds of formula (I) as defined above that can be used as antibiotics. Certain, they can be used to treat animals and humans with endoparasitic, ectoparasitic and / or fungal infections and in agriculture, horticulture, or forestry as pesticides to combat insects, acarid and nematode pests. They can also be used generally as pesticides to control or control pests in other circumstances, for example, in warehouses, buildings or other public places or locations of pests. Generally the compounds can be applied either to the host (animal or human or plants or vegetation) or to its location or to the pests themselves.

The compounds of the invention may be formulated for administration in any suitable manner for use in veterinary or human medicine and therefore the invention includes within its scope pharmaceutical preparations comprising a compound of the invention adapted for use in veterinary or human medicine. Such preparations may be adapted for use in a conventional manner by the use of one or more suitable ingredients or carriers. The compositions of the invention include those in the form of particularly formulated for parenteral (including intradermal administration), oral, rectal, topical, intraruminal, implantation of ocular, nasal or genito-urinary use.

The compounds of the invention may be formulated for use in veterinary or human medicine by injection and may be formulated in unit dosage form, in ampoules or other unit dose containers, or in multi-dose containers, if necessary with added preservative. Injectable compositions may be in the form of suspensions, solutions or emulsions, in non-aqueous or aqueous vehicles, and may contain formulating agents such as suspending, stabilizing, emulsifying, dissolving and / or dispersing agents. Alternatively, the active ingredient may be in the form of a sterile reconstitution powder

- 8 showering with a suitable carrier, for example, sterile pyrogen-free water, before use. Oily carriers include polyhydroxyl alcohols and their esters such as glycerol esters, fatty acids, vegetable oils such as walnut oil, cottonseed oil, mineral oils such as liquid paraffin, isopropyl myristate and ethyl oleate and other similar compounds. Other carriers may also be used that contain such materials as glycerol-formaldehyde, propylene glycol, polyethylene glycols, ethanol, or glyofural. Conventional non-ionic, cationic or anionic surfactants can be used alone or in combination in a preparation.

Veterinary preparations may also be formulated as intrauterine preparations in the form of bases or with rapid or slow action and may be sterile solutions or suspensions in aqueous or oily carriers which optionally contain a thickening or suspending agent such as soft or hard paraffins, bees wax,

12-hydroxy-stearin, hydrogenated castor oil, aluminum stearates or glyceryl monostearate. Conventional non-ionic, cationic or anionic surfactants may be used alone or in combination in a preparation.

«

The compounds of the invention may also be formulated for veterinary or medical use in a form suitable for oral administration, for example, in the form of solutions, syrups, emulsions or suspensions, or as a dry powder for reconstitution with water or other suitable carriers and before use, optionally with odorants and colorants. Takcd; solid preparations such as tablets, capsules, large tablets, pills, boluses, powders, pastes, granules, grains or premix preparations may be used. solid and liquid oral preparations may be made according to methods well known in the art. Such preparations may also contain one or more pharmaceutically acceptable carriers and ingredients, which may be either in solid or liquid form. Examples of suitable pharmaceutically acceptable carriers for use in the form of solid dosage units include binders (e.g., pregelatinized corn starch, polyvinylpyrrolidone or hydroxypropyl methylcellulose); fillers (e.g., lactose, microcrystalline cellulose or calcium phosphate); lubricants (e.g., magnesium stearate, talc or silica); chopping agents (e.g., potato starch or sodium starch glycolate); or wetting agents (e.g., sodium lauryl sulfate). The tablets may be coated by methods well known in the art.

Examples of suitable pharmaceutically acceptable additives for use as liquid dosage units include suspending agents (e.g., sorbitol syrup, methylcellulose or hydrogenated edible oils); emulsifying agents (e.g., lecithin or acacia); non-aqueous carriers (e.g., almond oil, oily esters or ethyl alcohol); and preservatives (e.g., methyl or propyl-p-hydroxybenzoates or sorbent acids); and stabilizing and solubilizing agents may also be included.

Oral administration pastes can be formulated according to methods well known in the art. Examples of suitable pharmaceutically acceptable additives for use in paste formulations include suspending agents or gelling agents, e.g., aluminum distearate or hydrogenated castor oil; dispersing agents, e.g., polysorbates; non-aqueous vehicles, e.g., walnut oil, oily esters, glyc or macrogels; stabilizing and solubilizing agents. The compounds of the invention may also be administered in veterinary medicine by their incorporation into solid or liquid daily meals of animals, e.g., as part of daily animal food or drinking water.

The compounds of the invention may also be administered orally in veterinary medicine in the form of such a liquid as a solution, suspension or dispersion of the active ingredient together with a pharmaceutically acceptable carrier or ingredient.

The compounds of the invention may, for example, be formulated for topical use, for use in veterinary and human medicine, such as ointments, creams, lotions, shampoos, powders, sprays, immersion agents, droplets (e.g., drops for the eyes or nose) or as a filler. Ointments and creams may, for example, be formulated with non-aqueous or oily basorine with the addition of suitable thickening and / or gelling agents. Eye grease can be produced in a sterile manner using sterilized components. The fillers may, for example, be formulated for veterinary use in organic solvents or as an aqueous suspension and may include percutaneous absorption promoting agents and formulation agents; which dissolve, stabilize, preserve or otherwise improve the storage properties and / or ease of use.

Lotions may be formulated with an aqueous or oily base and will generally also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents, thickening agents or coloring agents.

The powders may be formed by means of a suitable powder base (powder. The droplets may be formulated with an aqueous or non-aqueous base which also comprises one or more dispersing agents, stabilizing agents, solubilizing agents or suspending agents. They may also contain preservative.

For administration by inhalation, the compounds of the invention may be delivered for use in veterinary or human medicine in the formulation *, aerosol spray or inhaler.

The total daily doses of the compounds of the invention used in veterinary and human medicine will be suitably in the range of 1-2000 micrograms / kg body weight, preferably 5-800 g / kg and these may be administered in divided doses, e.g. e.g., 1-4 times daily. It will be clear that the dose will vary depending on the age and condition of the patient, the organism being treated, the route of administration and the particular formulated preparation. Doses for a particular patient can be determined using conventional considerations, e.g., by comparing the activity of the compound in question and that of a known antibiotic agent.

The compounds of the invention can be formulated in any convenient way for horticultural and agricultural use and the invention therefore includes within its scope preparations comprising a compound of the invention adapted for horticultural or agricultural use Such formulations include dry or liquid types, for example powders, including ordinary powders or concentrates, powders, including soluble or yeast powders, granules, including microgranules and dispersing granules, pills, liquid preparations, such emulsions as diluted emulsions or emulsifying concentrates, seed pills, seed fillings, immersion preparations such as preparations for root or seed immersion, oil concentrates, injections, e.g., injections for petioles, sprays, fumes and fogs.

Generally, such formulations will include the compound together with some suitable carrier or diluent. Such carriers may be liquid or solid and are intended to aid the administration of the compound or its dispersion when it is to be administered or to provide a formulation that can be made by the user himself in the form of a dispersing composition. Such formulations are well known in the art and can be made using conventional methods such as, for example, co-mixing and / or grinding the active ingredient (s) with a carrier or diluent, e.g., with a solid carrier , solvent or surfactant.

Suitable solid supports, for use in formulations such as powder; granules and powders may be selected, for example, from natural mineral fillers such as diatomite, talc, kaolinite, montmorillonite, profilite or atapulgite. Highly dispersed silicic acid or highly dispersed adsorbent polymers may be included in the preparation if desired. Granular adsorptive supports that can be used can be porous (such as float, milled tile, sepiolite or bentonite) or non-porous (such as calcite or sand). Suitable pre-granular materials which may be used and which may be organic or inorganic include dolomite and ground vegetable residues.

Suitable solvents for use as carriers or diluents include aromatic hydrocarbons, aliphatic hydrocarbons, alcohols and glycols or their ethers, esters, ketones, acid amides, strongly polar solvents, optionally epoxidized vegetable oils and water.

Conventional non-ionic, cationic or anionic surfactants, e.g., ethoxylated alkylphenols and alcohols, alkaline-metal or alkaline-earth metal salts of alkylbenzene-sulfonic acids, lignosulfonic acids or sulfo-amber acids, or sulfonates having good phenolic sulphonates and / or wetting can also be used either alone or in combination in a preparation.

Anti-baking agents, anti-foaming agents, anti-foaming agents, viscosity regulators, binders and adhesives, photostabilizers and fertilizers, food stimulants or other active substances, if desired, may be included in the preparations. The compounds of the invention may also be formulated in admixture with other insecticides, akari12 cides and nematicides.

In the formulations, the concentration of active material is generally from 0.01 to 99% and more, preferably between 0.01% and 40% by weight.

Commercial products are generally provided as concentrated prep. rates which should be diluted to an appropriate concentration, for example, from 0.001 to 0.0001% by weight, before use.

The rate at which a compound is administered depends on a number of factors including the type of pest involved and the degree of infection. However, mainly .; the application rate is 10 g / ha to 10 kg / ha; preferably from 10 g / ha to 1 kg, to control mites and insects and from 50 g / ha to 10 kg / ha to control nematodes.

The compounds of the invention may be administered or used in combination with other active ingredients. Specifically, the compounds of the invention may be administered or used in combination with other known anthelmintic agents. By combining the compounds of the invention with other anthelmintic agents, the spectrum of successfully parasitic infections can be expanded. Thus, it can be possible to eliminate parasitic infections against which individual components are ineffective or only partially effective.

The compounds obtained by the process of the invention may in some cases, during the process, have a protected hydroxyl group in the 5-position. Conventional procedures are used to protect and remove this protecting group, for example, as described in Green and McOmie's manuals.

The process of the invention for the preparation of a compound of formula (I) involves the reaction of a compound of formula (II):

(in which H _{2 is} NOR ² , or and, according to formula (I), formation with a salt thereof (wherein R is as desired, followed by deprotection in which OR ^{2 is a} protected hydroxyl group of the salt.

en som sano early edjer.ja optionally fine with ή

The oxidation reaction may be carried out in aqueous or non-aqueous reaction media, preferably at a temperature in the range of -20 to + 100 ° C, e.g., -10 to + 50 ° C. It is advantageous to use an H ₂ NOR ^Z reagent in the form of salts, for example, addition salts with an acid such as chloride. When used, such a reaction can be carried out in the presence of an acid-binding agent.

Solvents that may be used include water and quay solvents miscible with water such as alcohols (e.g., methanol or ethanol), amides (e.g., N, N-dimethylforraamide, Ν, Ν-dimethylacetamide or hexamet phosphoramide ), ethers (e.g., cyclic ethers such as tetrahydrofuran dioxane, and acyclic ethers such as dimethoxyethane or diethyl ether), nitriles (e.g., acetonitrile), sulfones (e.g., sulfolane), a hydrocarbon. such as halogenated hydrocarbons (e.g. methylene chloride) and esters such as ethyl acetate, as well as mixtures of two or more of such cancers.

When aqueous conditions are used, the reaction may be suitably buffered to pH 2-9 with a suitable acid, base or buffer.

Suitable acids include mineral acids such as hydrochloric or sulfuric acid, and such mineral acids as acetic acid. Suitable bases include alkali metal carbonates and bicarbonates such as sodium bicarbonate, hydroxides such as sodium hydroxide, and carboxylates of an alkali metal such as sodium acetate. A suitable buffer is sodium acetate / acetic acid.

The compounds of formula (II) are either known compounds as described in UK Patent Application 2176182 or can be made from known compounds as described herein using standard procedures.

The process of the invention may further comprise making 2 compounds of formula (I) in which R C is alkyl or a C 1-6 alkenyl group and OR is a substituted hydroxyl group, wherein 3 is a compound of formula (I) in which OR is a hydroxyl group reacted with a suitable reagent which is used to convert the hydroxyl group into a substituted hydroxyl group, followed by salt formation if necessary.

The acylation, formylation, sulfonylation, etherification, salylation or acetal formation reactions may be performed by conventional methods as described below.

Thus, for example, acylation can be carried out using such an agent.

for acylation such as an acid of the formula R COOH or a reactive derivative thereof, such as an acyl halide (e.g., acid chloride), an anhydride or activated ester, or a reactive derivative

4 carboxylic acids R OCOOH or thiocarboxylic acids R OCSOH.

Acylations using acid halides and anhydrides may optionally be carried out in the presence of an acid-binding agent such as a tertiary amine (e.g., triethylamine, dimethylaniline or pyridine), inorganic bases (e.g., potassium carbonate or sodium carbonate) ), and oxides such as some of the lower 1,2-alkylene oxides (e.g., ethylene oxide or propylene oxide) that bind the hydrogen halide released in the acylation reaction.

Acylations using acids are preferably carried out in the presence of a condensing agent, for example, a carbodiimide such as Ν, Ν′-dicyclohexylcarbodiimide or N-ethyl-N-gamma-dimethylaminopropylcarbodiimide; a carbonyl compound such as a carbonyldiimidazole; or some isoxazolium salts such as N-ethyl-5-phenylisoxazolium perchlorate.

An activated ester may conveniently be formed in situ using, for example, 1-hydroxybenzotriazole in the presence of a condensing agent as shown above. Alternatively, the activated ester may be reformed.

The acylation reaction may be carried out in aqueous or non-aqueous reaction media, preferably at a temperature in the range of -20 ° to + 100 ° C, e.g., -10 ° to + 50 ° C.

Formulation can be carried out using an activated formic acid derivative, e.g., N-formyl-imidazole or form-acet-anhydride, under standard reaction conditions.

The sulfonylation may be carried out with some reactive sulfonic acid derivative R ⁶ SO ₇ H such as some sulfonyl halide, for example, some 6 SCRCl chloride or some sulfonic anhydride. The sulfonylation is preferably carried out in the presence of a suitable acid-binding agent as described above.

The etherification can be carried out using a reagent of the formula R ^D y (wherein r 5 is as previously defined and Y represents a leaving group such as a chlorine, bromine or iodine atom or a hydrocarbylsulfonyl oxy group, such as mesyloxy or tosyloxy, or a haloalkanoyl oxy groups such as dichloroacetoxy). The reaction may be carried out by forming some magnesium alkoxide using such a Grignard reagent such as a methyl magnesium halide, e.g., methyl magnesium iodide, or using a trialkylsilylmethylmagnesium halide, e.g.

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 or mixtures thereof, and this system may be particularly appropriate when etherification is carried out using an alkyl halide (e.g. , methyl iodide).

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

Acetal formation can be accomplished by reaction with some cyclic or acyclic vinyl ether. This process is particularly useful for the manufacture of tetrahydropyranylether, using dihydropyran as a reagent, or such 1-alkoxyalkyl ethers such as 1-ethoxyalkylethyl, using alkylvinyl ether as a reagent. The reaction is preferably carried out in the presence of some strongly acid catalyst, for example, some such mineral acids such as sulfuric acid, or some organic sulfuric acids such as p-toluenesulfonic acid, in a non-hydroxyl solvent substantially free of water.

Solvents that can be used in the above reactions include ketones (e.g., acetone), amides (e.g., N, N-dimethylformamide, Ν, Ν-dimethyl acetamide or hexamethylphosphoramide), ethers (e.g., cyclic ethers such as tetrachydrofuran or dioxane, and acyclic ethers such as dimethoxyethane or diethyl ether, nitriles (e.g. acetonitrile), such hydrocarbons as sho are halogenated hydrocarbons (e.g. methylene chloride), and such esters such as ethyl acetate, as well as mixtures of two or more such solvents.

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

Carbamoylation to provide a compound of formula (I) wherein 3 is 8 9

The OC group OCONR R may be reacted with some suitable acylating agent (i.e., carbamoylation). Suitable carbamoylating agents which can be used to obtain the compounds in which it is eaten

9 of R and R is the hydrogen atom and the other is C.. alkyl groups include iso10-10 ^1-4 cyanates of the formula R NCO (where R is a ^C 1-4 alkyl group). The carbamoylation reaction may preferably be carried out in the presence of a solvent:

solvent mixtures selected from hydrocarbons (e.g. aromatic hydrocarbons such as benzene and toluene), halogenated hydrocarbons (e.g. dichloromethane), amides (e.g. formamide or dimethylformaraid), esters (e.g. e.g., ethyl acetate), ethers (e.g., cyclic ethers such as tetrahydrofuran and dioxane), ketone (e.g. acetone), sulfoxide (e.g. dimethylsulfoxide), or mixtures of these solvents. The reaction may conveniently be carried out at a temperature between -50 ° C and a boiling point of the reaction mixture, for example, up to 100 ° C, preferably between -20 ° and + 30 ° C.

Carbamoylation may be aided by the presence of a base, e.g., a certain tertiary amine such as tri- (loweralkyl) amine (e.g. triethylamine).

Another useful carbamoylating agent is cyanic acid, which is conveniently generated in situ, for example, from an alkali metal cyanate such as sodium cyanate, whereby the reaction is facilitated by the presence of an acid, e.g., some strong acid such as triflu. orosuric acid. Cyanic acid effectively corresponds to the isocyanate compounds mentioned above where R1 is hydrogen and therefore translates the compounds of formula (III) directly into their carbamoyloxy analogs (i.e., compounds of formula (I) in which OR1 is an OCONH ₂ group).

Alternatively, the carbamoylation may be carried out by reaction with phosgene or carbonyldiimidazole and, subsequently, with ammonia or with an appropriately substituted amine, optionally in an aqueous or non-aqueous reaction medium.

The formation of a compound of formula (I) in which OR represents the group OCO (CH ₂ ) _n CO ₂ R 4 can be achieved by acylation of the corresponding 5-hydroxy compound with an acid HO ₂ C (CH ₂ ) _n CO ₂ R 4 or with its reactive derivative according to the acylation method described above.

The process of the invention may also include, for the preparation of 2 compounds of formula (I) in which R 6 is alkyl or a C 2-6 alkenyl group, the reaction of a compound of formula (I) in which R 3 is a hydrogen atom and OR is a substituted hydroxyl group, with agent for 2 2 the etherification of RY (wherein Rc is C 1-6 alkyl or C 3-8 alkenyl group and

Y has the foregoing meaning), and subsequently, if necessary, the removal of 3 protection in a compound of formula (I) in which the OR is a protected hydroxyl group, and finally, if necessary, salt formation.

The etherification reaction may be carried out, for example, by the formation of magnesium zium alkoxide using some Grigr.ard reagent, such as methylmagnesium halocrenide, e.g., methylmagnesium iodide, followed by treatment with reagent R Y. Alternatively, the reaction may be to be carried out in the presence of some silver salts such as silver oxide, silver perchlorate, silver carbonate or silver salicylate or mixtures thereof, or in the presence of a base, for example, potassium carbonate or naurium hyd. The etherification may conveniently be carried out in an organic solvent such as an ether, e.g., diethyl ether, tetrahydrofuran or dioxane or an amide, e.g., dimethylformamide or hexamethylphosphoric triamide, or in a mixture of such solvents at ordinary temperature.

Under these conditions, the configuration of the oximino group is substantially unchanged by the etherification reaction.

Acid salts of formula (I) can be made by conventional methods, for example, by treating an acid with some basor or by converting one salt to another by ion exchange.

The invention is illustrated but not limited to the following Preparations and Examples in which the temperatures in ° C, 'L' are liter and EtOH is ethanol.

In the following Preparations and Examples, the compounds are referred to as derivatives of known 'Factors', Factors A, B, C * and D. Factor A is a compound

3 of formula- (VI) wherein R is isopropyl and R is hydrogen; Factor B is a compound of formula (VI) wherein R ^{1 is} methyl and R ¹ is methyl; Factor C

3 is a compound of formula (VI) wherein R is methyl and R is hydrogen; i 1 3

Factor D is a compound of formula (VI) wherein R is ethyl and R is hydrogen.

(VI)

Making 1- 5-Keto Factor A

Strsptomvcss tnermoarchensis spores NCIB 12015 inoculate ss on agar slant plates with the following ingredients

Yeast extract (Oxoid L21) 0.5 Malt extract (Oxoid L39) 30.0 Mycotic peptone (Oxoid L40) 5.0 Agar Ko. 3 (Oxoid L13) 15.0 Descilled water to 1 L pH about 5.4 and incubated at 28 ° for 10 days.

The mature plate is then coated with 6 ml of 10% glycerol solution and treated with sterile tools to loosen spores and also micelles. Aliquots of 0.4 ml of the resulting spore suspension were transferred to sterile polypropylene vessels, which were then sealed by heating and stored in liquid nitrogen vapor until needed.

Two Erlenmeyer 250 ml balloons containing 50 ml pelvic media consisting of:

gL ¹ D-Glucose 15.0 Glycerol 15.0 Soy peptone 15.0 NaCl 3.0 CaCO, 1 .0

The pH of the substrate was 6.7 and was adjusted to pH 7.0 with aqueous sodium hydroxide prior to autoclaving. The pH of the substrate after autoclaving was 7.3 / inoculated with 0.2 ml of spore suspensions taken from the vessel.

The balloons were inoculated at 28 ° for 3 days on a shaker rotating at 250 rpm with a 50 mm orbital diameter.

The contents of both balloons are used to inoculate a 70 L fermentor vessel containing 40 L of the same medium supplemented with polypropylene 2000 {0.06% v / v). Polypropylene 2000 was added as needed throughout the fermentation to control foaming. Fermentation is carried out at 28 °, with agitation and aeration satisfactory to maintain dissolved oxygen levels above 30% saturation. After 24 hours fermentation, a portion of the 9 L broth is transferred to a 700 L fermenter containing 450 L of substrate consisting of the following ingredients

D-Glucose 2.8 Malt Dextrin (MD3OE) 27.8 Arkasoy 50 13.9 Molasses 1.7 Κ, ΗΡΟ. 2 4 0.14 CaCO β 1 .39 Silicone 525 (Dow Corning) 0.06% (v / v) Adjust to pH 6.5 before sterilization. Fermentation is carried out at 28 with stirring and aeration. Polypropylene 20 ( antifoam is added as needed and pH is maintained at pH 7.2 added ·

by H ₂ SO ^ until harvest. The fermentation is harvested after 5 days.

The broth (450 L) was clarified on a Westfalia KA 25 centrifuge and the residual supernatant was replaced with water (20 L). The regenerated cells (25.5 kg) were mixed for 1 hour with a Silverson mixer Model BX in enough methanol to give a total volume of 75 L. The suspension was filtered and the solid was re-extracted with methanol (35 L) and filtered. The combined filtrate (87 L) was diluted with water (40 L) and extracted with petroleum ether at 60 ° -80 ° (30 L). After 30 minutes, the phases were separated on a Westfalia MEM 1256 centrifuge and the lower methanol phase was re-extracted with petroleum ether at 60-80 (30 L) after addition of water (40 L). After separation, the lower phase is again extracted with light petroleum at 60 ° -80 ° (30 L). Petroleum ether compounds (85 L) were concentrated with three passes through Pfaudler

8.8-12V-27 coated film evaporator (steam pressure 0.1 bar, steam temperature 20 °, hot steam temperature 127 °). The concentrate (9 L) was dried with sodium sulfate (2 kg) and further concentrated under reduced pressure to 4οθ in a rotary film evaporator.

The oily residue (130 g) was dissolved in chloroform to give 190 ml and this was applied to a Merck 7734 silica 60 (200x4 cm) column packed in chloroform. The column was washed with chloroform (500 ml) and eluted with chloroform: ethyl acetate (3: 1) and fractions of approximately 40 ml were collected after a pre-fraction of 1,400 ml.

Fractions 32-46 were combined and evaporated to give an oil (21.2 g). Fractions 47-93 were combined and evaporated to give an oil (20.1 g) which was dissolved in chloroform: ethyl acetate (3: 1) up to 50 ml, and applied to a Merck 7734 silica 60 (200x4 cm) column which was packaged in chloroform: ethyl acetate (3: 1), and fractions of approximately 40 ml were collected and evaporated to give an oil (3.1 g) which was added to an oil obtained from fractions 32-46 from the first column. The combined oils were dissolved in boiling methanol (4 ml) which was then added to the hot propan-2-ol (20 ml) to allow crystallization.

The mother liquor after crystallization is evaporated to give an oil which is dissolved in an equal volume of methylene chloride and batched on a column (30x2.2 cm) of Merck Kieselgel 60 (70-230 blend ASTM, Art. No. 7734) methylene chloride. The layer was washed with methylene chloride (2 volumes) and eluted with chloroform: ethyl acetate (3: 1) (2 layers). Evaporation of the eluate gave an oil which was dissolved in methanol and subjected to high performance preparative liquid chromatography (hplc) on Spherisorb S5 ODS-2 (250nunx20mm, Phase Sep. Ltd.). Sample batches are pumped onto the column for a period of 1 minute, the column is eluted with acetonitrile: water (7: 3) under the following conditions:

eme (min.) Flow rate (mil / min) 0.00 o.00 (Time 1.00 0.00 injection) 1.10 30.00 39. 90 30.00 40.00 35.00 75.00 35.00

The material eluted from the hplc column was followed by UV spectroscopy at 238 nm.

Evaporation of the combined fractions with peaks eluting at 33.4 minutes afforded the title compound (34 mg) as a solid.

E.I. mass spectroscopy gave the molecular ion at 610 and gave the characteristic fragments at: 592

574

556

422

259

241 example 1

23 / E / -Methoxyimino Factor A, 5-acetate

A solution of anhydrous sodium acetate (2.8 g) in water (15 ml) was added to a solution of 23-keto Factor A, 5-acetate (3.13 '^ g, Example 18 in UK Patent Application 21761 82) in methanol, and then, and methoxyamine chloride (3.01 g). The resulting solution was mixed with 0.5N hydrochloric acid, water and brine. The dried organic phase was evaporated to near dryness and the off-white foam was purified by chromatography over Merck Kieselgs 60 230-400 mixture (600 ml). Elution of the column with hexane: ethyl acetate (4: 1) afforded the title compound as a colorless foam (2.14 g) / alpha / _D + 128 ° (c 1.35, CHCl 2) lambda (EtOH) 244 nm (epsilon 27,250);

· “J ΓΠ & Χ ΙΠ3.Χ ni (CHBr-,) 3560, 3480 (OH), 1733 (acetate), 1715 (C = O), delta (CDC1,) ΓΠ, & Χ 3 3 includes 5.5-5.6 (m; 2H ), 3.84 (S; 3H) 3.29 (d 15; H), 2.16 (S; 3H).

Example 2

23 / E / -Hydroxyimino Factor A, 5-acetate

The reaction of 23-keto FactorA, 5-acetate with hydroxylamine hydrochloride was carried out in a similar manner to that described in Example 1 above.

The crude product was purified by chromatography over Merck Kieselgel 60

230-400 was stirred, eluting with ethyl acetateacetonitrile (4: 1) to give the title compound as a colorless foam / alpha / + 132 ° (c 1.01,

CHC1-), lambda (EtOH) 244 nm (epsilon 27800), ni (CHBr _o ) max max max 3

3565, 3470 (OH), 1732 (acetate), 1712 (C = 0), 993 (CO), delta (CDC1 ₃ ) included 8.12 (S; 1H), 5.5-5.6 (m; 2H), 3.42 (dl 5 1H), 2.16 (S; 3H).

Example 3

23 / E / -Methoxyimino Factor A

A solution of the product of Example 1 (1.88 g) in methanol was cooled in an ice bath, 1N aqueous sodium hydroxide (5.6 ml) was added and the solution was stirred in an ice bath for 1.5 hours. The solution was diluted with ethyl acetate and washed successively with 0.5N aqueous hydrochloric acid, water and brine. The dried organic phase was evaporated and the resulting foam was purified by chromatography over Merck Kieselgel 60 230-400 mixture (400 ml). Elution columns with hexane: ethyl acetate (2: 1) gave a colorless foam (1.429 g). Crystallization from hexane gave the pure title compound, mp 203 °, / alpha / ^{1 *} 132 + (c 1.21, CHCl 2), lambda (EtoH) 244 nm (epsilon 29200), ni (CHBr-,) 3540 (OH ), max ^c max 'max 3

1708 (C = O), 992 (C-O), aeltaiCDCl-1) included 4.29 (t7: 1H), 3.84 (s; 3H), 3.29 (d15; 1H).

Example 4

23 / E / -Hydroxyimino Factor A

Hydrolysis of the product of Example 2 according to the procedure described in

Example 2 above gave a product which was purified by chroinatography over Merck Kieselgel 60 230-400 mesh (400 ml) eluting with hexane:

ethyl acetate (1: 1) to give the title compound as a colorless

Dena / alpha / ¹ + 140 ° (c 1.24, CHC1-), lambda _m _ (EtOH) 244 nm (epsilon 26700) ni (CHBr,) 3565, 3490 (OH), 1710 (C = O), 994 max max j (CO), deltafCDCl2) included 8.11 (S; 1H), 4.29 (t7; 1H), 3.41 (d15; 1E

Example 5

23 / E / -Ethoxyimino Factor A

A solution of anhydrous sodium acetate (140 mg) in water (3 ml) was added to a solution of 23-Keto Factor A (200 mg, Example 23 in the U.K. Patent

Report 2176182) and ethoxyamine hydrochloride (126 mg) in methanol (20 ml)

After 2 hours at 20 ° the solution was diluted with ether (40 ml) and washed with water. The dried organic phase was evaporated and the resulting off-white foam was purified by chromatography over Merck Kieselgel 60 230-400 mixture (90 ml). Elution of the column with hexane: ethyl acetate (2: 1) gave the title compound as a colorless foam (189 mg) / alpha / + 125 ° (c 1.00, CHCl 3 lambda (EtOH) 244 nm (epsilon 28,200) or (CHBr,) 3540 , max max 'max 3

3480 (OH), 1705 (C = 0), 990 (CO), delta (CDCl ₃ ) included

4.30 (t, 1H), 4.10 (q7; 2H), 3.31 (d15; 1H), 1.24 (t7; 3H).

The compounds of Examples 6, 7 and 8 were similarly made from 23-keto Factor A and the corresponding alkoxyamine.

Example 6

23 / E / -Alyloxyimino Factor A / alpha / p ¹ + 124 ° (c 1 .17, CHCl 2), lambda _max (EtOH) 244 nm (epsilon ^^ 28,400), ni (CHBr,) 3550, 3490 (OH ), 1708 (C = O), 990 (CO) delta (CDCl ₃ ) includes 5.98 (m; 1H), 5.28 (dd17.2; 1H), 5.15 (dd9.2; 1E) 4.5-4.7 (m; 2H) ), 4.29 (t7; 1H), 3.36 (d14; 1H) is made from allyloxyamine hydrochloride.

Example 7

23 / E / -Isopropyloxyimino Factor A / alpha / ^{* 21} + 116 ° (c 0.97, CHC1.J, larnbda _{m = iv} (EtOH) 244 nm (epsilon 'D - 3 max max

25,000), neither (CHBr) 3550, 3490 (OH), 1708 (C = O), 992 (CO), delta max (CDC1 ₃ ) includes 4.2-4.4 (m; 2H), 3.30 (d14; 1H), 1.21 (d7; 3H), 1.20 (d7; 3H) is made from isopropyloxyamine hydrochloride.

Example 8

23 / E / -n-Butoxyimino Factor A / alpha / ²¹ + 115 ° (c 1.10, CHC1,), lambda (EtOH) 244 nm (epsilon 'D - 3 max * max

31,800), nor (CHBr ^) 3540, 3460 (OH), 1708 <C = O), 992 (CO), max 3 delta (CDC1 ₃ ) includes 4.28 (t6; 1H), 4.03 (m; 2H), 3.96 (d6; 1H),

3.31 (d14; 1H), 0.9-1.1 (m; 15H) is made from n-butoxyamine hydrochloride.

Example 9

23 / E / -Methoxyimino Factor A, 5-acetate (1) A 3-molar solution of methyl magnesium iodide in ether (0.16 ml) was added to a stirred solution of the product of Example 2 (120 mg) in dry hexamethylphosphoric triamide (5 ml) under nitrogen. Iodomethane (0.09 ml) was added after 1 hour, the mixture was diluted with ethyl acetate (30 ml) and washed successively with 2N hydrochloric acid and water. The dried organic phase was evaporated and the yellow gum was purified by chromatography over Merck

Kieselgel 60 230-400 blends (80 ml). Elution of the column with hexane: ethyl21 o Daddy (2: 1) gave the title compound as a white foam / alpha / _D + 123 ^ (c 1.25, CHC1_) lambda (EtOH) 245 nm (epsilon 30,300). The NMR was as described in Example 1.

(ii) The product of Example 2 (0.082 g) was dissolved in diethyl ether (10 ml) containing silver oxide (0.4 g), freshly made from aqueous silver silicate and 2M sodium hydroxide). The mixture was stirred at room temperature for an hour, after which it was filtered and the solvent was evaporated to give a crude yellow gum. The residue was purified by preparative thin layer chromatography (Merck 5717) eluting with dichloromethane / acetone (25: The title bar was extracted with acetone and evaporated to give the title compound (0.059 g). NMR was as described above in Example 1 ·

Example 1θ

23 / E / -Methoxyimino Factor A, 5-methylcarbamate

Methylisocyanate (0.13 ml) (125 mg) and triethylamine (2 drops) were added n. a solution of 23 (E) -methoxyimino Factor A (350 mg) in dry dimethylformamic (0.75 ml). The balloon was inflated and heated for 5.5 hours at 80 ° with stirring. The reaction mixture was poured into water (50 ml) and the resulting mixture filtered through kieselguhr. The cake on the filter was washed with water (150 ml) and then extracted with dichloromethane (75 ml). The extract was dried (MgSO4) and concentrated to give a yellow foam which was purified by column chromatography on silica (125 mg, Merck Kieselgel 60, 230-4CC stirred) under medium pressure. Elution with hexanesethyl acetate (1: 1) gave the title compound as a white foam (206 mg). [alpha] + + 99 ° (c 0.55, CK ₃ C1 _O ); lambda (EtOH) 244.4 nm (epsilon 26710); ni - zz max max (CHBr ₃ ) 3530 (OH), 3455 (NH), 1720 (ester), 1720 + 1510 (carbamate) and 993 cm ¹ (C-0); delta (CDCl 2) included 1.78 (s, 3H), 2.86 (d, 5Hz, 3 3.29 (d, 14Hz, 1H), 3.83 (s, 3H), 4.80 (g, 5Hz, 1H) and 5.50 (m, 2H) ).

Example 11

23 / E / -Methoxyimino Factor A, 5-methylcarbonate ϋ solution of 23 / E / -methoxyimino Factor A (150 mg) in dichloromethane (15 ml) and pyridine (0.3 ml) was added methyl chloroformate (0.7 ml of 1.0M solution in dichloromethane) with stirring. The reaction mixture was allowed to stir at 0-3 ° for 20 minutes, then added to dichloromethane (70 ml) and washed with 2N hydrochloric acid (50 ml) and water (50 ml). The organic phase was dried (MgSO4) and the solvent was separated to give a foam which was purified by chromatography on a column of silica (40 g,

Merck kieselgel .60, 230-400 mixes) under medium pressure. Elution with dichloromethane: ethyl acetate (30: 1) gave the title compound as a white foam (127 mg), [alpha] + ¹ + 145 [deg.] (C = 0.41, CH _O C1 _O ); lambda at £, z rnax (EtOH) 244.4 nm (epsilon 31210); ni (CHBr,) 3460 + 3540 (OH),

ΙΗαΧχ J

1742 (carbonate) 1710 (ester) and 992 xm (C-0); delta (CDCl2) include 1.82 (s, 3H), 3.29 (d 14 Hz, 1H), 3.82 (s, 3H), 3.83 (s, 3H), 5.2-5.4 (m; 3H) 5.56 (s, 1H) .

Example 12

23 / E / -Methoxyimino Factor D, 5-acetate

Solution containing 23-keto Factor D, 5-acetate (251 mg, Example 119 u

U. S. Patent Application 2176182), sodium acetate (250 mg) and methoxyimin chloride (250 mg) in methanol (40 ml) is held at 20 ° for 24 hours, concentrated to about 10 ml, diluted with ethyl acetate (50 ml) and washed is successively with 0.5N hydrochloric acid and water. The dried organ was evaporated to give a yellow foam which was purified by chromatography over Merck Kieselgel 60, 230-400 mesh (120 ml). Eluting the column with hexane gave the title compound as a light yellow foam (144 mg);

lambda (EtOH) 244 nm (epsilon 26,400); ni (CHBr _n ) (cm ^Ί ) 3500 max max □ (OH), 1732 (OAc), 1710 (C = O); delta (CDCl3) included 5.54 (m; 2H), 4.92 (m; 1H), 3.84 (s; 3H); 3.32 (m; 1H), 3.30 (d14; 1H), 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 13.

23 / E / -Methoxyimino Factor B

A solution containing 23-keto Factor B (1g, Example 19 in UK Patent Application 2176182), sodium acetate (400 mg) and methoxyamine chloride (400 mg) was stirred at 20 ° for 20 hours, concentrated to about 10 ml. , diluted with ethyl acetate and washed with water. The organic phase was washed successively with 0.5N hydrochloric acid and water, and the dried organic phase was evaporated and the crude product was purified by chromatography over Merck Kieselgel 60, 230-400 stirred (200 ml). Elution of the column with ethyl acetate: chloromethane (1: 9) gave the title compound as a white foam (500 mg); / alpha / i? ¹ + 128 ° (c 1.09, CHCl 3); larnbda _mav (EtOH)

244 nm (epsilon 30,100); not _in (CHBr,) (cm *) 3540, 3460 (OH),

1708 (C = O); delta (CDCl3) included 5.46 (q6; 1H), 4.03 (d5; 1H),

3.97 (d5; 1H), 3.83 (s; 3H); 3.50 (s; 3H), 3.32 (m; 1H), 3.29 (d14; 1H), 1.82 (s; 3H), 1.68 (d6; 3H), 1.00 (d6; 3H), 0.92 (d6; 3H) .

Example 14

23 / E / -Methoxyimino Factor C

Anhydrous sodium acetate (0.54 g) and methoxyamine hydrochloride (0.5cg) were added to a solution of 23-keto Factor C (1.97 g, Example 12 in U.K. Patent Application 2176182) in methanol (30 ml) containing water (5 ml). the mixture was stirred for 30 minutes at room temperature. Ethyl acetate (30 ml) and 0.5M hydrochloric acid were added and the aqueous layer was re-extracted with ethyl acetate (15 ml). The combined organic layers were washed again with 0.5M hydrochloric acid, with 5% saturated aqueous sodium bicarbonate and 10% saturated aqueous sodium chloride, then concentrated in vacuo to a yellow foam which was purified by chromatography on Merck 9385 silica gel, initially the column was developed with dichloromethane and then eluted with dichloromethane containing a small amount of ethyl acetate (up to 10%) to give the title compound (1.0 g); [alpha] + + 64 ° (c 1.0, CH 2 OH); NMR (CDCl3) included the following signals: delta 4.95 (m, 1H); 4.29 (t, 1H, 7Hz); 3.96 (d, 1H, 7 Hz); 3.85 (s, 3H / = NOCH ₃ /); 3.66 (d, 1H, 10 Hz); 1.51 (s, 3H); 1.42 (t, 1H,

12Hz); IR (CHBr ₃ ) 3620-3340 cm ^-1 (-0H), 1711 cm ¹ (C = O).

The following are examples of formulations according to the invention. The term 'Active ingredient' as used hereinafter means a compound of the invention and may optionally be a compound of Example 3.

Multiaise parenteral injection

Example 1

Active ingredient

Benzyl alcohol

Polysorbate 80

Glycerol-formal

Water for injections up to% m / v Interval 2.0 0.1-6.0% w / v .0

10.0

50.0

100.0

Dissolve the active ingredient in polysorbate 80 and glyceroliormal. Add benzyl alcohol and make up to volume with water for injections. Sterilize the product by conventional methods, for example, by sterile filtration or by autoclaving, and pack aseptically.

Example 2

Active ingredient Benzyl alcohol Glycerol triacetate Propylene glycol ao% w / v

4.0

2.0

30.0

100.0

Interval 0.1 - 7.5% w / v

Dissolve the active ingredient in benzyl alcohol and glycerol triacetate. Add propylene glycol and make up to volume. Sterilize the product with conventional pharmaceutical methods, for example, by sterile filtration and pack aspetically.

Example 3

Active ingredient Ethanol Non-ionic surfactant (e.g. Svnperon; Propylene glycol

2.0 m / v 36.0 m / v PE L44 *) 10.0 m / v to 100.0 ingredient in ethanol and.

Interval

0.1 -7.5% w / v volumes. Sterilize product with conventional pharmaceutical processes, for example, sterile filtration and aseptically packaged, * ICI Trade name

Example 4

Active ingredient

Non-ionic surfactant (e.g., Svnperonic PE F68 *) Benzylalcohol

Miglyol 840 **

Water for injections up to 100.0

2.0 m / h

2.0 m / v 1.0 m / v

16.0 v / v interval 0.1 --3.0% w / v

Dissolve the active ingredient in Miglyiol 840. Dissolve the non-ionic son factor and benzyl alcohol in most of the water. Make an emulsion

- 30-. by adding an oily solution to the aqueous solution with homogenisation using conventional means. Make up to volume. Aseptically prepare and pack aseptically.

* ICI trade name ** Dynamit Nobel trade name

Aerosol spray

Active ingredient Trichloromethane Trichlorofluoromethane Dichlorodifluoromethane

% m / m Interval 0.1 0. G1 - 2.0% w / w 29.9 35.0 35.0

Mix the active ingredient with trichloroethane and fill in an aerosol container. Inflate the end space with water propellant and put the valve in position. Fill the required weight of the liquid propellant under pressure through the valve. Store with actuators and caps.

Tablet

Production process - wet granulation

Ξ2

Active ingredient 250.0 Magnesium stearate 4.5 Corn starch 22.5 Sodium starch glycolate 9.0 Sodium lauryl sulfate - 4.5

Microcrystalline cellulose to the inside weight of the tablet

450 mg

Add a satisfactory amount of 10% starch paste to the active ingredient to produce a suitable wet granulation mass. Prepare the granules and dry them using a drier with a tray or with a fluid layer. Siphon through a sieve, add remaining ingredients and compress into tablets.

If necessary, coat the inside of the tablets with a film using hydroxypropyl methylcellulose or other similar material that forms the film using either an aqueous or non-aqueous solvent system. The plasticizer and a suitable color may be included in the film forming solution.

Veterinary pill for use on small / domestic animals Production process - dry granulation mg

Active ingredient 50.0

Magnesium stearate 7.5 * ·

Microcrystalline cellulose for the inside of a 75.0 tablet

Mix the active ingredient with magnesium stearate and microcrystalline cellulose. Put the mixture in a lump. Crush the lumps by passing through a rotary granulator to produce fine mobile granules. Compound into tablets.

The inner parts of the tablet can be film coated, if desired, as described above.

Veterinary intradermal injection mg / dose

150 mg

Active ingredient Polysorbate 60 3.0% w / w))

White beeswax 6.0% w / w) up to 3 g) up to 3 or 15 g Walnut oil 91.0% w / w)

Interval 0.05 - 1.0 g

Heat the walnut oil, white beeswax and polysorbate 60 to 16 ° C with stirring. Maintain at 160 ° C for two hours and then cool to room temperature with stirring. Aseptically add the active ingredient to the carrier and disperse using a high speed mixer. Puffin by passing through a colloidal mill. Aseptically fill the product in sterile plastic syringes.

Slight Release Veterinary Bolus% m / m Interval,

Active ingredient) 0.25-2g

Colloidal silicon dioxide 2.0) is filled as needed

Microcrystalline cellulose up to 100.0)

Mix the active ingredient with colloidal silica and microcrystalline cellulose using a suitable aliquot mixing technique to achieve a satisfactory distribution of the active ingredient throughout the entire honeycomb. Insert into the light release device and adjust to (1) the constant release of the active ingredient or (2) the pulse release of the active ingredient.

Veterinary oral

Active ingredient Polysorbate 85 Benzyl alcohol Propylene glycol Phosphate buffer Water liquid% w / v

0.35

5.0

3.0

30.0 as pH 6.0 to 100.0

Interval 0.01 - 2% w / v

6.5

Dissolve the active ingredient in Polysorbate 85, benzyl alcohol and propyl glycol. Add a batch of water and adjust to pH 6.0-6.5 with phosphate buffer, if necessary. Make up to volume with water. Fill the product into a liquid container.

Veterinary oral paste

Active ingredient

Sodium saccharin

Polysorbate 85

Aluminum um-distearate. Fractionated Coconut Oil% m / m 4.0 2.5 3.0 5.0 to 100.0

Interval 1-20% w / w

Disperse aluminum distearate in fractionated coconut oil and polysorbate 85 by heating. Cool to room temperature and disperse sodium saccharin in an oil carrier. Disperse the active ingredient into the base. Fill in plastic syringes.

Granules for veterinary administration in food% m / m

Active ingredient 2.5

Calcium sulfate, hemi-hydrate up to 100.0

Interval 0.05 - 5% w / w

Mix active ingredient with calcium sulfate. Prepare granules using the wet granulation process. Dry with a drier or a fluid bed. Fill in appropriate container.

Pouring Veterinary Nrenate - ---------- - - - * ·· -. -. Active ingredient

Dimethylsulfoxide

Methylisobutylketone

Propylene glycol (and pigment) up to% m / v Interval

2.0 0.1 to 30%

10.0

30.0

100.0

Dissolve the active ingredient in dimethylsulfoxide and methylisobutyl ketone. Add pigment and make up to volume with propylene glycol. Screw in a container for pouring preparation.

Emulsifying concentrate

Active ingredient 50 g

Anionic emulsifier 40 g (e.g. Phenyl sulphonate CALX)

Non-ionic emulsifier 60 g (eg Synperonic NP13) *

Mix all ingredients and mix until dissolved.

* Trade name ICI

Granules (a) Active ingredient 50 g

Wood resin 40 g

Gypsum granules (20-60 meshes) up to 1 kg (eg Agsorb 100A) (b) Active ingredient 50 g

Synperonic NP13 * 40 g

Gypsum granules (20-60 meshes) up to 1 kg.

Dissolve all ingredients in a volatile solvent, e.g., methylene chloride, add granules by shaking in a mixer. Dry to separate solvent.

* Trade name ICI

The pesticidal activity of the compounds of the invention was determined by various pests and their hosts according to the following general procedure:

The product is useful as a liquid preparation. The preparations were made by dissolving the product in acetone. The solutions were then diluted with water containing 0.1% or 0.01% by weight. wetting agents until the liquid preparations contain the required amount of product.

The accepted treatment procedure for many pests included the support of a large number of pests on the rootstock, which was usually a native plant and either the substrate was treated with the preparation (residual test) or, in the case of Tetranychus urticae, Mvzus persicae, Nilaparvata lugens and Mus ca domestica. and pests and substrate treated with the preparation (contact test). In the case of Meloidogna incognita, the solution was applied to the soil in which the tomato plants were grown for, was subsequently treated with nematodes and the reduction in the number of nodes at the root was evaluated in comparison with the control plant.

By the following methods, the compound of formula (I) in 12 3 to which R is isopropyl, R is methyl and R is hydrogen is found to be effective at concentrations (by product weight) of 100 parts per million or less.

THE APPLICANT'S STATEMENT ON THE BEST KNOWLEDGE WAY FOR THE ECONOMIC USE OF THE APPLICATION FOUND

23 / E / -Methoxyimino Factor A

A solution of the product of Example 1. (1.88 g) in methanol was cooled to ice:.

· 1N aqueous sodium hydroxide (5.6 ml) was added and the solution stirred in an ice bath for 1.5 h. The solution was diluted with ethyl acetate and washed successively with 0.5N aqueous hydrochloric acid, in:

and brine. The dried organic phase was evaporated and the resulting foam was purified by chromatography over Merck Kieselgel 60 230-400 mixture (400 ml). Elution columns with hexane; ethyl acetate (2: 1) gave a colorless foam (1.429 g). Crystallization from hexane gave the pure title compound, mp 203 °, / alpha / ¹ + 132 ° (c 1.21, CHCl-j), lambda (EtoH) 244 nm (epsilon 29200), ni (CHBr,) 3540 (OH) , max ^r max 'max 3 ·

Similarly, by reacting 23-keto Factor A, 5-acetate with hydroxylamine hydrochloride, 23 / E / -hydroxyimino Factor A is obtained,

5-acetate.

Claims (1)

PATENT APPLICATION A process for the preparation of novel macrolide compounds of general formula (I) wherein R ¹ represents a methyl, ethyl or isopropyl group; R ² represents a hydrogen atom, an alkyl group or a C ₃ ^ alkenyl group and the group = NOR ² in E-configuration; OR ³ is a hydroxyl group or a substituted hydroxyl group having up to 25 carbon atoms and pharmaceutically acceptable salts thereof, characterized in that it comprises reacting compounds of formula (II) with H ₂ NOR ² reagent or a salt thereof (R ¹ , R ² and OR ³ are as defined above), and then, where necessary, to prepare compounds of formula (I) wherein R ^{2 is an} alkyl or C 3-6 alkenyl group and OR ^{3 is a} substituted hydroxyl group, the reaction of compounds of formula (I) is carried out , wherein OR ^{3 is a} hydroxyl group, with a reagent to convert a hydroxyl group into a substituted hydroxyl group, or to prepare compounds of formula (I) wherein R ^{2 is an} alkyl group and a C 3 ^ alkenyl group, the reaction of compounds of formula (I) is carried out, Iger is a R ² hydrogen atom or an OR ³ substituted hydroxyl group, with an esterifying agent R ² Y (where R ^{2 is an} alkyl and C ₃ ^ alkenyl group and Y is a leaving group), and then optionally removing the protecting group from the protected hydroxyl group OR ³ in the compound of the form lo (I) and the compounds of formula (I) are optionally converted into their salts.