LT3684B - Macrolide antibiotics and process for preparing thereof - Google Patents

Abstract

Compound and salts of the same are represented by the formula (II)<IMAGE> (II)whereR<1> marks methyl, ethyl or isopropyl group:R<2> marks hydrogen atom or OR<5> group (where OR<5> is a hydroxyl group or changed hydroxyl group having up to 25 carbon atoms) and R<3> marks hydrogen atom or R<2> and R<3> together with a carbon atom to which they are being connected marks )C=O group; andOR<4> marks OR<5> that is defined above;under condition that, when R<2> marks hydroxyl group, OR <4> is changed hydroxyl group, but not a methoxyl group. These compounds may be used to fight against insects, ticks, nematodes and other pests.

Description

The present invention relates to antibiotic compounds and methods for their preparation.

United Kingdom patent application no. No. 8522699 We describe the production of S541 antibiotics, which can be isolated from fermentation products of a new species of Streptomyces. S541 antibiotics are a group of similar compounds of general formula (I).

OH

Another group of compounds having antibiotic activity, which may be obtained by modifying S541 antibiotics or isolating them from a culture of the microorganism of the genus Streptomyces, are provided herein. The novel compounds of the invention exhibit antibiotic activity and / or are used as intermediates for the preparation of other active compounds and / or for the isolation and purification of S541 antibiotic compounds.

The compounds of the present invention are analogs of the 23-ketone, 23-deoxy and 23-hydroxyl or substituted hydroxyl S541 antibiotics having a hydroxyl or substituted hydroxyl group at the 5-position.

Thus, in one aspect of the invention, the compounds represented by formula (II) are mainly obtained,

OR ⁴ . . 2 wherein R is methyl, ethyl or isopropyl; R is a hydrogen atom or an OR group (where OR is a hydroxyl group or a substituted hydroxyl group having up to 23 carbon atoms) and R is a hydrogen atom, or R and R ³ together with the carbon atom to which they are attached form> C = Group O; OR ⁴ corresponds to the group OR ⁵ defined in 2 above and their salts, provided that when R is a hydroxyl group, OR ⁴ is a substituted hydroxyl group but not a methoxyl group.

When the compounds of formula (I) are used as intermediates, one or both of the R ² and -OR ⁴ groups will most often be a protected hydroxyl group, and the invention relates specifically to such protected compounds.

When the R ² or OR ⁴ groups of the compounds of formula (I) are substituted hydroxyl groups, they may be the same or different and be an acyloxy group [e.g., the group of formula -OCOR ⁶ , -OCO ₂ R ⁶ or -OCSOR ⁶ (where R ⁶ is an aliphatic, arylfat or aromatic group (e.g., alkyl, alkynyl, cycloalkyl, arylalkyl or aryl)]. formyloxyl

7 6 group -OR group (wherein R corresponds to R as defined), -OSO ₂ R group (wherein R is a C ₃ _ ₄ alkyl or C ₆ _ ₁₀ aryl group), sililoksilo group, cyclic, acyclic 3684 B acetalLT oxy group or the group OCO (CH ₂ ) _n CO ₂ R ⁹ (where R ⁹ is a hydrogen atom or a group as defined above for R ⁶ , on may have the meanings 0, 1 and 2).

When R or R is an alkyl group such as a C _{x 8} alkyl group such as methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl or n-heptyl, these alkyl groups may also be substituted. When R ⁶ is a substituted alkyl group, it may be substituted, for example, by one or more, e.g., two or three halogen atoms (e.g., chlorine or bromine), or carboxyl, C 4 -C 4 alkoxyl (e.g., methoxyl, ethoxyl, phenoxyl, or siliioksilo group. If R ⁷ is a substituted alkyl group, it may be changed C3_ ₇ cycloalkyl, eg., cyclopropyl group.

When R ⁶ or R ⁷ is an alkenyl or alkynyl groups, they may be eg. C ₂ _ ₈ alkenyl such as allyl, or C ₂ _ ₈ alkynyl groups.

When R ⁶ or R ⁷ is a cycloalkyl group, they may eg. C ₃ _ ₁₂ cikloalkiias, such as C ₃ _ ₇ cikloaikilas. Thus, R ⁶ may be, for example, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. R ⁷ can be, for example, a cyclopentyl group.

When R ⁶ or R ⁷ are arylalkyl groups, they usually have from 1 to 6 carbon atoms in the alkyl region, and the aryl group (s) may be carbocyclic or heterocyclic and most often are 4 to 15 carbon atoms, such as phenyl. Examples of such groups are phenyl C ₇ _ ₆ alkyl, such as benzyl or phenyl.

When R or R are aryl groups they may be carbocyclic or heterocyclic and usually have 4 to 15 carbon atoms, such as phenyl.

When R ² or -OR ⁴ is -OSO ₂ R ⁸ , it may be, for example, methylsulfonyloxy or p-toluenesulfonyloxy.

When R ² or -OR ⁴ represents a cyclic acetyloxy group, it may have, for example, 5-7 ring members and be, for example, a tetrahydropyranyloxy group.

When R ² or OR ⁴ represents a silyloxy group, or R ⁶ has a silyloxy substituent, the silyl group may combine three groups which may be the same or different from alkyl, alkenyl, alkoxyl, cycloalkyl, aralalkyl, aryl and aryloxy groups. The same may be defined as for R ⁶ and generally include methyl, t-butyl and phenyl. Specific examples of the silyloxy group include trimethylsilyloxy and t-butyldimethylsilyloxy.

When R ² or -OR ⁴ represent OCO (CH 2) n CO 2 R ⁹ group, it may be eg., OCOCO2R ⁹ OCOCH ₂ or CO ₂ R ^9, wherein R ⁹ is a hydrogen atom or an alkyl (eg. Methyl or ethyl) group .

Compounds of formula (II) containing an acidic group may form salts and the corresponding bases. Such salts include alkali metal salts such as sodium and potassium salts.

Important formula (II) group of compounds are those wherein R ¹ 2 4 represents methyl, ethyl or isopropyl group, R and R, which may be the same or different, each represents OR ⁵ (where OR ⁵ is a hydroxy group or substituted hydroxy group with up to 25 carbon atoms), and R ³ is a hydrogen atom, and salts with the proviso that when R ² is hydroxy, then OR ⁴ is a substituted hydroxyl group, but not a methoxyl group.

Another important group of compounds of formula (II) is that wherein R ¹ represents a methyl, ethyl or isopropyl group,

R ² and R ³ together with the carbon atom to which they are attached form a> C = O group and OR ⁴ is a hydroxyl group of OR ⁵ or a substituted hydroxyl group having up to 25 carbon atoms) and their salts.

Another important group of compounds of formula (II) is that wherein R ¹ represents a methyl, ethyl or isopropyl group, both R ² and R ³ are hydrogen and OR ⁴ is OR ⁵ (wherein OR ⁵ is hydroxy or substituted hydroxyl group containing up to 25 carbon atoms) and their salts.

In the compounds of formula (II), the group R ¹ is primarily an isopropyl group.

The group R is primarily a hydrogen atom or a hydroxyl group or a -OCOR ⁶ group of the formula [wherein R ⁶ is a C 3-8 alkyl group (a freely substituted C 4-4 alkoxyl group) or a phen C 1-6 alkyl group], -OCO ² R ⁶ (wherein R ⁶ is C 1-6 g). alkyl groups which are replaced by from one to three halogen atoms, such as., trichloroethyl) -OCOCO2H, -OR ⁸ (wherein R ⁸ is Ο ₄ _ ₈ alkyls, C ₃ _ ₇ cycloalkyl, allyl or cyclopropylmethyl group) and R ² and R ³ together with the carbon atom to which they are attached form a> C = O group. Importantly, R is primarily a hydrogen atom or an ethoxyl, n-propoxyl, cyclopropylmethoxyl, acetoxyl, phenacetoxyl, propionoxyl, isobutyrionoxyl or cyclopropanecarbonyloxy group or R and R together with the carbon atom to which they are attached form a> C = O group.

In the compounds of formula (II), the group -OR is primarily hydroxyl, methoxyl, acetoxyl or methyloxycarbonyloxy.

The active compounds of formula (II) according to the invention are those wherein R ¹ is methyl, ethyl and especially isopropyl, R is a hydrogen atom or an ethyloxy, n-propoxyl, acetoxyl or propionoxyl group, R is a hydrogen atom or R and R together with the carbon atom to which they are attached form a> C = O group, and OR ^{4 is a} hydroxyl, acetoxyl, or methoxycarbonyloxy group.

Of particular interest are those active compounds of formula (II) of the invention wherein:

3

R is an isopropyl group, R is a hydrogen atom, R is a hydrogen atom, and OR ⁴ is a hydroxyl group;

R is an isopropyl group, R is a propionoxyl group, R 4 is a hydrogen atom, and OR is a hydroxyl group;

R ¹ is an isopropyl group, R ² and R ³ taken together with the carbon atom to which they are attached form a> C = O group, and -OR is a hydroxyl group;

R ¹ is an isopropyl group, R ² is an ethoxyl group, R ³ is a hydrogen atom, and -OR ⁴ is a hydroxyl group; and

R is an isopropyl group, R is an n-propoxyl group, R is a hydrogen atom, and -OR ⁴ is a hydroxyl group.

R is a methyl group, R is an acetoxyl group, R is a hydrogen atom, and OR ⁴ is a hydroxyl group.

R is an ethyl group, R is an acetoxyl group, R is a hydrogen atom, and OR ⁴ is a hydroxyl group.

3

R is an isopropyl group, R is an acetoxyl group, R is a hydrogen atom, and OR ⁴ is a hydroxyl group.

As noted above, the compounds of the present invention may be used as antibiotics and / or as tarltic compounds to provide other active compounds and / or to isolate and purify S541 antibiotic compounds. Where the compounds of the invention are intended to be used as intermediates, the R ² and / or -OR ⁴ groups may be protected hydroxyl groups. It is understood that such a group should have a minimum number of additional functional groups in order to avoid other reaction chains, and should also be capable of selectively regenerating the hydroxyl group. Examples of protected hydroxyl groups are well known and described, e.g., Theodora W. Greene in Protective Groups in Organic Synthesis (WileyInterscience, New York 1981) and JFW McOmie in Protective Groups in Organic Chemistry (Plenum Press, London, 1973), R ² and examples of OR ⁴ with protected hydroxy groups are phenoxyacetoxyl, silyloxyacetoxyl (e.g. trimethylsilyloxyacetoxyl and t-butyldimethylsilyloxyacetoxyl), and silyloxy such as trimethylsilyloxy and t-butyldimethylsilyloxy. Compounds of the invention having such groups are used in particular as intermediates. Other groups, such as acetoxyl, may replace the protected hydroxyl group, although they may also be present in the final active compounds.

Other active compounds of the invention useful as intermediates are compounds of formula (II) wherein R is a -OCOCO ₂ H group. They are particularly suitable for isolation and purification of S541 antibiotic compounds. A compound of formula (II) wherein R ¹ is particularly useful in this regard. . R 2 is an isopropyl group, R is a -OCOCO ₂ H group, R is a hydrogen atom and OR is a hydroxyl group that can be successfully used to isolate and purify the antibiotic compound S541 of formula (V) below, wherein R ¹ is isopropyl, and R is a hydrogen atom. The crude compound of formula (V) provides the corresponding compound of formula (II) wherein

R ² is -OCOCO ₂ H and is isolated, for example, in crystalline form. The compound of formula (V) may then be recovered in substantially purified form from the latter compound using methods described herein, the isolation and purification methods of which form part of the present invention.

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

Ectoparasites and endoparasites infect humans and many animals, and are particularly prevalent in animals such as pigs, sheep, large horns, goats and birds, horses and domestic animals such as dogs and cats. Parasitic livestock infections causing anemia, malnutrition and weight loss are the leading cause of economic loss worldwide.

The genera of endoparasites infecting these animals and / or humans include Ancylostoma, Ascaridia, Ascaris, Aspicularis, Brugia, Bunostomum, Capilliara, Chabertia, Cooperia, Dictyocaulus, Dirofilaria, Dracunculus, Enterobius, Haemonchus, Heterakis, Loa,

Nematodirus, Nematospiroides (Heligomoroides), Nippostrongylus, Oesophagostomum, Onchocerca, Ostertagia, Oxyuris, Parascaris,

Strongyioides, Syphacia, Toxascaris,

Strongylus,

Toxocara,

Trichonema, Trichostrongylus, Uncinaria, and Vuchareria.

Trichineila, Trichuri,

Among the animal and / or human infectious ectoparasites are arthropod ectoparasites, which include stinging insects, meat flies, fleas, lice, mites, sucking insects, ixodic mites and other dicotyledonous pests.

The genera of ectoparasites infecting these animals and / or humans include Ambylomma, Boophilus, Chorioptes,

Culliphore, Demodex,

Gastrophilus, Haematobia,

Demallenia,

Dermatobia,

Haemophysalis,

Haematopinus,

Hyalomma, Hyperderma, Ixodes, Linognathus, Lucilia, Melophagus, Oestrus, Otobius, Otodectes, Psorergates,

Psoroptes,

Tabanus.

Rhipicephalus, Sarcoptes, Stomoxys, and

The compounds of the invention have been found to be effective against many endoparasites and ectoparasites both in vitro and in vivo. The compounds of the invention have been found to be particularly active against nematode parasites including Haemonchus contortus, Ostertagia circumcincta, Trichostrongylus colubiformis, Dictyocaulus viviparis, Cooperia oncophera, Ostertagia ostertagi, Nematospiroides dubius and Nippostrongylus, and Nippostrongylus. and Psoroptes sp.

Thus, the compounds of the invention are useful in the treatment of animals and humans with endoparasitic and / or ectoparasitic infections.

The parasite species varies with the host and the prevailing site of infection. Therefore, for example, Haemonchus contortus, Ostertagia circumcincta and Trichostrongylus colubiformis usually infect sheep and are usually localized in the stomach and small intestine, while Dictyocaulus viviparis, Cooperia oncophera and Ostertagia ostertagi tend to infect the large gastropod, and the common gastropod, respectively.

Antibiotic activity of the compounds of the invention in vitro may be demonstrated, for example, by their action against free-living nematodes such as Caenorhabiditis elegans.

In addition, the compounds of the invention are useful as antifungal agents, e.g., against Candida sp. strains such as Candida albicans and Candida glabrat'a and against yeasts such as Saccharomyces carlsbergensis.

The compounds of the invention are also useful in the control of pests, insects, mites and nematodes, agriculture, horticulture, forestry, public health and product retention. Soil pests and crops such as cereals (e.g. wheat, barley, maize and rice), vegetables (e.g., soy), fruits (e.g., apples, vines and citrus), and root crops (e.g. beets, potatoes) can be successfully processed. Typical representatives of such pests include fruit mites and aphids such as Aphis fabae, Aulacorthum circumflexum, Myzus persicae, Nephotettix cincticeps, Nilparvata lugens, Panonychus ulmi, Phorodon humuli, Phyllocoptruta oleivora, Tetranychus urtica, genera Aphelencoides, Globodera, Heterodera, Meloidogyne and Panagrellus; butterflies such as Heliothis, Plutella and Spodoptera; cereal thorns, such as Anthonomus grandis and

Sitophilus granarius;

Tribolium castaneum; domestica; Richter moths, such as flies, such as Musca _ ants; moth-worn narrow-wing; Pear psylla; Thrips tabaci; cockroaches such as Blatella germanica and Periplaneta americana and mosquitoes such as Aedes aegypti.

Thus, the present invention provides compounds of formula (II), as described above, which may be used as antibiotics. In particular, they can be used for the treatment of animals and humans with endoparasitic, ectoparasitic and / or fungal injections and as pesticides for killing insect pests, mites and nematodes in agriculture, planting or forestry. They can also be used as pesticides to combat pests and in other circumstances, such as in warehouses, buildings or other areas of the public or in pests. In general, these compounds can be applied to either hosts (animals or humans, crop plants or other vegetation) or to the pests themselves and their foci.

The compounds of the invention may be formulated for administration by any suitable route in veterinary or human therapy, and therefore the invention includes pharmaceutical compositions containing a compound of the invention for use in veterinary and human therapy. Such mixtures may be presented for administration by conventional means, together with suitable carriers or excipients. The compositions of the invention are specifically formulated for parenteral (including delivery to mammary gland), oral, rectal, topical application or implantation. Suitable methods and agents for preparing the compounds of the invention for use in veterinary or human medicine are described in South African patent no. 85/7049 on S541 Antibiotic Compounds.

The compounds of the invention may be administered in combination with other pharmaceutically active ingredients.

The total daily amount of the compounds of the invention used in both veterinary and human therapy will be in the range of 1-2000 µg / kg, preferably 10-1000 µg / kg, and more preferably 100-500 µg / kg, and may be administered in divided doses, e.g. 104 times a day.

The compounds of the invention may be formulated for use in any suitable manner in horticulture or agriculture, and therefore the invention includes mixtures containing a compound of the invention for use in horticulture or agriculture. Such formulations are of a dry or liquid type, such as dusts, which include dusty substrates or concentrates, powders, which contain soluble or wettable powders, granules, which contain micropellets and dispersible granules, tablets, volatiles, emulsions such as soluble emulsions or emulsifiable concentrates, disinfectants such as roots and seeds, fertilizers for seeds, tablets for seeds, oily concentrates, oily solutions, for example, stems, pulverulent liquids, smoke and mists.

Suitable methods and agents for preparing the compounds of the invention for use in horticulture or agriculture are described in South African patent no. 85/7049 on S541 Antibiotics.

In formulations, the concentration of active ingredient is usually from 0.01 to 99%, more preferably from 0.01 to 40% by weight.

Commercial products are usually supplied in the form of concentrated mixtures which are diluted to the required concentration, e.g. 0.001 to 0.0001% by weight, before use.

When used in veterinary or horticultural applications and in agriculture, and when the compounds of the invention are obtained from fermentation products, it is desirable to use the whole fermentation broth as a source of active compounds. It is also suitable to use dry broth (with micelle) or micelle separated from broth and pasteurized, or better still, dried. If necessary, the broth or micelle may be prepared in admixture with inert carriers, excipients or diluents as described above.

The antibiotic compounds of the invention may be administered or used in combination with other active ingredients.

In addition, the antibiotic compounds of the invention may be used in conjunction with the antibiotic compounds S541 or other antibiotic compounds of the invention. This occurs when, for example, the crude fermentation products are subjected to the process of the invention without prior or subsequent isolation; this may be acceptable in agriculture when it is important to maintain low production prices.

The compounds of the invention may be prepared by the methods discussed below. In some of these ways, it may be necessary to protect the hydroxyl group of the starting material at position 5 or 23 before initiating the reactions described. In cases where the reactions are carried out to obtain the desired compound of the invention, it is necessary to deprotect the hydroxyl group. For protection or removal, conventional techniques such as those described in Greene and McOmie can be used.

In this way, for example, an acyl group such as an acetyl group may be removed by basic hydrolysis, e.g., using sodium or potassium hydroxides in an alcoholic aqueous solution, or by acid hydrolysis, e.g., using a methanolic solution of concentrated sulfuric acid. Acetyl groups such as tetrahydropyranyl may be cleaved, for example, by acid hydrolysis (using these, acetic or trifluoroacetic acids, or dilute mineral acid). The filing groups may be cleaved using fluoride ions (e.g., tetralkylammonium fluoride, such as tetra-n-butylammonium fluoride), hydrogen fluoride in acetonitrile aqueous solution, or an acid, such as p-toluenesulfonic acid (e.g., methanol). The arylmethyl groups may be separated by treatment with, for example, lysic acid (e.g., boron trifluoride ether) in the presence of a thiol (e.g., ethanolic) in an appropriate solvent, e.g., dichloromethane, e.g., at room temperature. The selective deprotection at the 5-position in the 5,23-disilyl compound of the invention may be carried out using tetra-n-butylamine fluoride, and the selective deprotection at the 5,23-diacetoxy compound may be carried out using sodium hydroxide in aqueous methanol.

The compounds of the invention in which R ² and / or -OR ⁴ is a substituted hydroxyl group can generally be obtained by interaction of antibiotic compounds S541 (e.g. compounds of formula (V) below) or their 5- or 23-O-mono derivatives with the reagents used. for the formation of a substituted hydroxyl group. In general, the 5-hydroxyl group is more active than the 23-hydroxyl group.

The hydroxyl group at the 5-position is more easily deprotected than at the 23-position. In general, the 5-mono-substituted compounds of the invention are obtained by treating the 5,23-unsubstituted S541 antibiotic compound with a limited amount of reagents and under milder conditions, while the 5,23-unsubstituted compounds are obtained under less mild conditions using higher amounts of reagents and / or catalyst. The 23-mono-substituted compounds of the invention are obtained by first forming a 5,23-substituted compound and selectively deprotecting at the 5-position. 5-23-Substituted compounds of the present invention having different substituents at the 5- and 23-positions may be obtained by interaction of a mono-substituted or 5- or 23-position compound with a reagent used to form a different substituted hydroxyl group at another position.

According to a further aspect of the invention there is provided a process for the preparation of compounds of formula (II) for2 4 mules wherein one of R and OR is a substituted hydroxyl group and the other is a hydroxyl or substituted hydroxyl group as described above comprising a compound of formula (III).

OR ⁴ (III) (wherein -OR ⁵ is substituted by one of -OR and is hydroxyl or by reacting with a reagent, the group is replaced by hydroxyl

R is as previously defined to convert a hydroxyl group and another hydroxyl group into a hydroxyl group and, if necessary, with subsequent selective deprotection of a compound of formula (II) wherein both R ² and OR ⁴ are substituted hydroxyl groups, to give a compound of formula (II) wherein OR ⁴ is a hydroxyl group and R ² is a substituted hydroxyl group.

In general, acylation, formylation, sulfonylation, etherification, silylation reaction or acetal formation will occur.

Thus, for example, acylation may be carried out using an acylating agent such as an acid of formula R ⁶ COOH or an active derivative thereof, such as a halogen anhydride (e.g. chloro anhydride), anhydride, activated ester, R ⁶ COOH or monocarboxylic acid. thiocarbonic acid R ⁶ CSOH derivative.

Acylation using halogen anhydrides or anhydrides may be carried out, where appropriate, in the presence of an acid-binding agent such as a triple amine (e.g., triethylamine, dimethylline or pyridine), inorganic bases (e.g., calcium carbonate or sodium bicarbonate), and oxiranes, such as lower 1,2-alkylene oxides (e.g. ethylene or propylene oxide), which bind the hydrogen halogen liberated during the acylation reaction.

Acylation using strongly electrophilic chloro anhydrides (e.g., oxalyl chloride, methoxyacetyl chloride, chloroacetichloride or bromoacetyl chloride), preferably with an acid binder such as calcium carbonate, can be used to selectively convert 23-position wherein R is an OR group, wherein R is a substituted hydroxyl group and OR ⁴ is a hydroxyl group.

It is desirable that the acid acylation be carried out in the presence of a condensing agent such as a carbodiimide such as N, N'-dicyclohexylcarbodiimide or N-ethyl-γ-dimethylaminopropylcarbodiimide; a carbonyl compound such as carbonyldiimidazole; or an isoxazole salt such as N-ethyl-5-phenylisoxazole perchlorate.

The activated ester may be conventionally prepared in situ using, for example, 1-hydroxybenzotriazole in the presence of a condensing agent as noted above. Alternatively, the activated ester may be briquetting.

The acylation reaction can take place in an aqueous or nonaqueous reaction medium, typically at a temperature of -20 ° C to + 100 ° C, such as -10 ° C to + 50 ° C.

Formylation can be carried out using an activated formic acid derivative such as N-formyl imiLT 3684 B dazole or formic acid anhydride under standard reaction conditions.

Sulfonation can occur when the active sulfonic acid R ⁸ SO ³ H can be present, a sulfonyl halide such as chloride R ⁸ SO ² Cl or a sulfonic anhydride. Sulfonylation is preferably carried out when an appropriate amount of an acid-binding agent is present, as described above.

The etherification may be carried out using a Y reagent of formula R ⁷ (wherein R ⁷ is as previously defined and Y is a cleavable group such as a chlorine, bromine or iodine atom or a hydrocarbylsulfonyloxy group such as mesyloxy or tosyloxy or a haloalkanoyloxy group such as dichloroacetoxy). ). The reaction may take place with the formation of magnesium alcoholate using a Grignard reagent such as methylmagnesium halide, such as methylmagnesium iodide, or a trialkylsilylmethylmagnesium halide, such as trimethylsilylmethylmagnesium chloride, followed by the reaction with R ⁷ Y.

Alternatively, the reaction may take place in the presence of a silver salt such as silver oxide, silver perchlorate, silver carbonate or silver salicylate, or mixtures thereof, and the suitability of such a system is particularly evident when etherification is carried out using an alkyl halide (e.g., methyl iodide).

Etherification can be carried out routinely in solutions such as ethers such as diethyl ether.

Acetal can be formed by reaction with a cyclic or acyclic vinyl ether. This method is particularly good for the preparation of tetrahydropyranyl ethers using the dihydropyran reagent or 1-alkoxyalkyl ethers such as 1-ethoxyalkyl ether using the alkyl vinyl ether reagent. The reaction is preferably carried out in the presence of a strong acidic catalyst such as a mineral acid such as sulfuric acid or an organic sulfuric acid such as p-toluenesulfonic acid in a non-hydroxylic, substantially anhydrous solvent.

Anhydrous solvents such as ketones (e.g. acetone), amides (e.g. N, N-dimethylformamide, N, N-dimethylacetamide or hexamethylphosphoramide), ethers (e.g. cyclic ethers, e.g. such as tetrahydrofuran or dioxane, and cyclic 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, as well as mixtures of two or more such solvents.

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

In many cases, e.g., when the 5,23-dihydroxy compound of formula (III) is used as starting material, a mixture of end products, such as a mixture of 5-mono-substituted and 5,23-disubstituted derivatives, will be obtained. They can be separated by conventional techniques such as chromatography such as silica chromatography or high performance liquid chromatography.

In another embodiment of the invention, a compound of formula (II) wherein R and R represent one hydrogen atom may be obtained by interaction of a compound of formula (III) wherein R is as previously defined and OR is a substituted hydroxyl group and R ⁵ is a hydroxyl group. obtained by substitution of the 23-hydroxyl group with a hydrogen atom and deprotection of the OR ⁴ group if junLT 3684 B is required, wherein OR ⁴ is a hydroxyl group, followed by further substitution for the compound of formula (II) in which OR ⁴ is a substituted hydroxyl group.

Thus, the starting materials of formula (III) wherein OR is a substituted hydroxyl group and OR ⁵ is a hydroxyl group can interact with a reagent having a hydroxyl group replaced by a cleavage group to give compounds of formula (IV):

OR ⁴ [wherein R ¹ and OR ⁴ are just defined and L is an atom or group that is cleaved by reduction, for example., Homolitinės reduction time, such as R ⁿ OCSO- [wherein R ^u is Ο ₄ _ ₆ alkyl, aryl such as phenyl, or ₍₇ Ο _ ₆ alkyl) aryl, such as p-toluenoksitiokarboniloksilo group]].

Suitable reagents that can be used to introduce the moiety L include, for example, aryl haloenzymatic formates such as p-tolyl chlorothioformate. The reaction may take place in the presence of a base, e.g. an amine such as pyridine when the solvent is a hydrocarbon halide such as dichloromethane.

If necessary, intermediates of formula (IV) may be isolated.

The intermediates of formula (IV) may then be reduced to the desired compounds of formula (II) using a reducing agent such as an alkyl tin hydride (e.g., tri-n-butyl tin hydride) in the presence of a radical initiator such as peroxide, azobisisobutyrnitrile or light.

The reaction can usually be accomplished by the choice of a solvent between a ketone such as acetone; an ether such as dioxane, a hydrocarbon such as hexane or toluene; a hydrocarbon halide such as trichlorobenzene; or an ester such as ethyl acetate. Mixtures of several solvents, alone or with water, may also be used.

The reaction may be carried out at a temperature of 0 ° to 200 ° C, preferably 20 ° to 130 ° C.

In another embodiment of the invention, a compound of formula (II) wherein R ² and R ³ together with the carbon atom to which they are attached form a> C = O group can be obtained by oxidation of a compound of formula (III) wherein OR is a hydroxyl group , and OR is a substituted hydroxyl group, followed by deprotection of the OR group if a compound wherein OR ⁴ is hydroxyl is required, and if necessary re-substituted to provide a compound of formula (II) wherein OR ⁴ is substituted hydroxyl group. The reaction may take place in the presence of an oxidizing agent which must convert the second hydroxyl group to an oxo group, resulting in the compound of formula (II).

Suitable oxidizing agents include quinones in the presence of water, such as 2,3-dichloro-5,6-dicyano-1,4-benzoquinone or 2,3,5,6-tetrachloro-1,4-benzoquinone; chromium (VI) oxidizing agent, e.g. pyridine dichromate or chromium trioxide pyridine; a manganese oxidizing agent such as manganese dioxide in dichloromethane; An N-halosucinimide such as N-chlorosuccinimide or N-bromosuccinimide; a dialkylsulfoxide such as dimethylsulfoxide in the presence of active agents, N, N'-dicyclohexylcarbodiimide or an acyl halide such as oxalyl chloride; or sulfur-pyridine trioxide complex.

The reaction can proceed successfully by the choice of a solvent between a ketone such as acetone; an ether such as diethyl ether, dioxane or tetrahydrofuran; a hydrocarbon such as hexane or toluene; a halide of hydrocarbons such as chloroform or methylene chloride or an ester such as ethyl acetate or a substituted amide such as dimethylformamide. Mixtures of several solvents, alone or with water, may also be used.

The reaction can be carried out at temperatures between -80 ° C and + 50 ° C.

According to a further aspect of the invention there is provided a process for preparing a compound of formula (II) wherein R and R together with the carbon atom to which they are attached form a> C = O group and OR ⁴ is a hydroxyl or methoxy group comprising Streptomyces genus. culturing the organism capable of producing at least one compound of the invention and, if necessary, isolating said compound, as well as, if necessary, modifying the OR ⁴ group by the methods described above.

Proposed microorganisms capable of producing the above materials are strains of new species of the genus Streptomyces, which have been named Streptomyces thermoarchaensis. A sample of this microorganism, a soil isolate, was deposited (September 10, 1984) with a permanent culture collection at the National Industrial and Marine Bacteria Collection, Torry Research Station, Aberdeen, United Kingdom, and was given accession number NCIB 12015. Streptomyces thermoarchaensis The NCIB 12015 mutants can also be used successfully, and 4 mutant strains were deposited (June 26, 1985) into a permanent culture collection at the National Industrial and Marine Bacteria Collection under accession numbers NCIB 12111, NCIB 12112, NCIB 12113 and NCIB 12114. .

The compounds of the invention may be prepared by conventional methods of Streptomyces organisms, e.g. culturing Streptomyces organisms, when sources of carbon, nitrogen and mineral salts are assimilated into the medium.

The assimilable sources of carbon, nitrogen and mineral salts can be either simple or complex substances. Carbon sources will typically consist of glucose, maltose, starch, glycerol, molasses, dextrin, lactose, sucrose, fructose, carboxylic acids, amino acids, glycerides, alcohols, alkanes, and vegetable oils. The carbon sources will, as a rule, be 0.5 to 10% by weight of the fermentation medium.

Nitrogen sources will typically include soy flour, wheat liquids, distillate solutions, yeast extracts, cotton seed meal, peptones, peanut flour, malt extract, molasses, casein, amino acid mixtures, ammonia (gas or liquid), ammonium salts or nitrates. In addition, urea and other amides may be used. The nitrogen sources will, as a rule, be 0.1 to 10% by weight of the fermentation medium.

The nutrient mineral salts which may be included in the culture medium will consist of the common salts of sodium, potassium, ammonium, iron, magnesium, nickel, cobalt, manganese, vanadium, chromium, calcium, copper, molybdenum, boron, phosphorus, sulphate and chlorine. carbonate ions.

Streptomyces organisms are cultured generally at 20 to 50 ° C, preferably 25 to 40 ° C, and preferably about 34 ° C, and are preferably aerated and shaken, e.g., by shaking or stirring. Initially, a small amount of sporulated suspension of microorganisms may be inoculated, but avoiding retardation of growth, the growing crop may be prepared by inoculating organisms in the form of spores into a small amount of nutrient medium and the resulting growing crop may be transferred to a fermentation medium. , allowing for further growth, one or more passages before transfer to the main fermentation medium. The fermentation usually takes place at a pH range of 5.5 to 8.5, preferably 5.5 to 7.5.

The fermentation can take from 2 to 10 days, for example about 5 days.

If the entire fermentation broth is to be isolated with the compounds of the invention or any single compound can be isolated, then conventional separation and isolation techniques are used. The compounds of the invention are predominantly present in the cell micelle but can also be detected in the fermentation broth, so that the isolation techniques can also be applied to the fermentation broth after clarification. It is understood that quite different methods of separation may be chosen.

Various fractionation methods, such as adsorption, precipitation, fractional crystallization and solvent extraction, may be used to isolate and isolate the compounds of the invention, which may be variously combined.

Solvent extraction and chromatography have been found to be most suitable for the separation and isolation of the compounds of the invention.

After fermentation, the micelle can be collected by conventional methods such as filtration and centrifugation. Subsequently, the appropriate organic solvent, such as ketones, e.g. acetone, methylene ketone or methyl isobutyl ketone, may be used to extract the compounds from the micelle; hydrocarbon, e.g. hexane; hydrocarbon halides such as chloroform, carbon tetrachloride or methylene chloride; alcohols such as methanol or ethanol; or esters, such as methyl acetate or ethyl acetate. It is understood that if the micelle is aqueous, it is preferable to use aqueous solvents.

In general, extraction is performed more than once for optimal compound recovery. For the first extraction, it is preferable to use water miscible solvents such as methanol or acetone. Antibiotics can be extracted as crude extracts by removing the solvent. Solvent extracts, in turn, can be extracted by reducing the volume of the solvent, for example by evaporation. Water-immiscible solvents such as hexane, chloroform, methylene chloride, or ethyl acetate or mixtures thereof are more suitable at this stage, in addition to the addition of the requisite amount of water for better material distribution. Removal of the water immiscible phase yields a product containing one or more compounds of the invention, most probably S541 antibiotic compounds.

The purification and / or separation of the compounds of the invention can be carried out by conventional techniques such as chromatography (including high performance liquid chromatography, hplc) with an appropriate sorbent such as silica, non-functional large adsorbent resin, e.g. polystyrene resin. cross-links such as Amberlite XAD-2, XAD-4 or XAD-1180 (Rohm & Haas Ltd) resins, or S112 resin (Dash Ltd or organic solvent miscible dextran such as Sephadex LH20 (Pharmacia UK Ltd)) or HPLC the case of a reversed phase adsorbents such as hydrocarbon linked silica eg. C ₁₈ -surištas silica. The sorbent may be trough-shaped, and even better - filled column. Non-functional stambiatinklės resin such as XAD- 1180 or S112, organic solvents such as acetonitrile with water are suitable for leaching.

Silica and Sephadex columns are usually filled with a solution of the compounds and the appropriate solvent and, if necessary, reduced by volume of the solvent. The column can be optionally rinsed and subsequently rinsed with solvent of the required polarity. In the case of silica and sephadex, solvents such as alcohols such as methanol may be used; hydrocarbons such as hexane, acetonitrile; halide hydrocarbons such as chloroform or methylene chloride; or esters such as ethyl acetate. Combinations of solvents, either alone or with water, may also be used.

The leaching and purification / purification of the compounds of the invention can be controlled by conventional techniques such as thin layer chromatography and high performance liquid chromatography or by using the properties of the described compounds.

I

The compounds of the invention may first be purified by silica chromatography, preferably using a similar chloroform eluent: ethyl acetate, followed by, for example, high performance liquid chromatography. The resulting purified material can then be chromatographed on a sephadex column with an eluent such as acetonitrile, and then the compounds of the invention can be isolated by high performance liquid chromatography.

After the compound is obtained in substantially pure form, it is further purified by crystallization, e.g. from methanol, isopropanol or methanol / water; the invention also encompasses those compounds of the invention which are in crystalline form.

Following appropriate procedures, the compounds of formula (II), as hereinbefore defined, were isolated in solid form. It will be appreciated that the sequence of purification steps, their choice and the degree of purification sought may vary considerably. As described above, compounds having a purity level consistent with the intended use may be used. When used in human therapy, a purity of the compounds of less than 90%, preferably greater than 95%, is desirable. A lower purity, such as 50% or less, will be suitable for veterinary or other purposes.

The intermediates of formula (III) of antibiotic S541, wherein OR ⁵ is a hydroxide group and OR ⁴ is a hydroxyl or methoxyl group, may also be prepared using the fermentation and isolation methods described above, e.g. 85/7049. Other intermediates of formula (III) may be prepared from these compounds using the methods described above for the preparation of compounds of formula (II).

The following procedures and examples illustrate the invention. Temperatures are indicated in ° C.

The compounds are named with reference to the Factors which are the compounds of formula (V) below

The factor R R ¹ A -H -CH (CH ₃ ) ₂ B -ch ₃ -ch ₃ C -H -ch ₃ D -H -ch ₂ ch ₃ E -ch ₃ -ch ₂ ch ₃ F -ch ₃ -CH (CH ₃ ) ₂ Factors A, B, C, D, E, F, and E may be obtained

South African patent no. 85/7049.

example

5-Phenoxyacetoxy and 5,23-Diphenoxyacetoxy Factor A

Factor A (2.0 g) in dichloromethane (25 mL) and pyridine (0.35 mL) was treated with a solution of phenoxyacetyl chloride (0.5 mL) in dichloromethane at 0 °. After 18 h at 3 °, the solution was treated with pyridine (1.0 mL) and phenoxyacetyl chloride (1.0 mL) in dichloromethane (5 mL). The solution was stirred for 30 min before being added to ice water. At 0-5 °. After addition of ether (100 mL), the mixture was stirred for 20 min. The aqueous layer was extracted with ether. The ether layers were mixed, washed successively with water and brine, dried and evaporated. Purification of the residue by chromatography on a silica column using dichloromethane: acetone (40: 1) afforded a mixture of the title compounds (1.8 g, monoacyl: diacyl = 6: 1) which, after application, reversed-phase preparative hplc. to obtain 5-phenoxyacetoxy Factor A, δ (CDCl ₃ ) should be 6.8 to 7.4 (m; 5H) and 4.66 (s;

2H); m / z has 746, 728, 710, 594, and 576, and 5.23-diphenoxyacetoxy Factor A, δ (CDCl ₃ ) has from 6.8 to 7.4 (m, 10H), 4.60 (s, 2H), and 4.70 (s, 2H).

example

5-Phenoxyacetoxy-23- (4-methylphenoxithionecarbonyloxy) Factor A

5-Phenoxyacetoxy Factor A (747 mg) in dichloromethane (10 mL) at 0 ° C under nitrogen was treated with pyridine (0.81 mL) followed by 4-methylphenyl chlorthionformate (0.75 g) in dichloromethane (2). ml). The resulting dark solution was stirred for 15 min. 0 ° C and then 22 hours without refrigeration. The solution was poured into cold water and brine, then extracted with ether. The combined ether layers were washed with water and brine, dried and evaporated. Purification of the residue by chromatography on silica and reverse phase preparative hplc gave the title compound (430 mg), δ (CDCl ₃ ) having 3, 34 (m; 1H), 3, 58 (m,

1H), 3.97 (d10; 1H), 4.72 (s; 2H), 5.4 (m; 1H), 5.59 (d6; 1H), and 6.9 to 7.4 (m; 9H) m / z has 728, 616, 576, 466, 464, 448, 354, 297, 247, 219 and 151.

example

5-tert-Butyldimethylsilyloxyacetoxy Factor A

Factor A (2.144 g) in dry ether (25 mL) and pyridine (2.5 mL) was treated dropwise with t-butyldimethylsilyloxyacetyl chloride (1.2 g) in ether (10 mL) at 0 ° C. The mixture was stirred for 90 min. At 0 ° C, acid chloride (1.10 g) in ether was added dropwise thereto. The mixture was stirred for 60 min. At 0 ° C and poured into cold water and ether. The aqueous layer was washed with ether. The combined organic layers were washed with water and brine, dried and evaporated. Purification of the residue by silica gel column chromatography using dichloromethane: acetone (25: 1) gave the title compound δ

(CDC1 ₃₎ is 0.09 (s, 6H), 0.78 (d6; 3H), 0.90 (s, 9H), 0.93 (d6; 3H), 0.97 (d6; 3H), 1.03 (d6; 3H), 1.51 (s; 3H), 1.59 (s, 3H), 1.74 (s, 3H), 3.32 (m; 1H), 3.52 (d10; 1H) , 3.64 (m; 1H), 3.74 (d10, 1H), 3.82 (m; 1H), 4.32 (s; 2H), 5.75 (d5; 1H), m / z has 784, 766, 748, 595, 577, 484, 466, 354, 314, 297, 265, 247, 237, 219,

and 151.

example

5-tert-Butyldimethylsilyloxy Factor A

Factor A (250 mg) and imidazole (163 mg) in purified dimethylformamide (10 mL) were treated with t-butyldimethylsilyl chloride (197 mg). The solution was stirred for 2 hours. and poured into cold water. The mixture was thoroughly extracted with ether and the mixture of ethereal extracts was dried and evaporated. The residue was purified by chromatography on silica gel using dichloromethane: acetone (10: 1) as the eluent, the title compound was obtained (235 mg), δ (CDC1 ₃₎ is 0.13 (s, 6H),

0, 80 (d6; 3H), 0, 9: 2 (s, 9H), 0, 96 (6d; 3H), 1.00 (d6; 3H), 1.03 (d6; 3H) , 1.53 (s; 3H), 1.60 (s; 3H), 1.80 (s; 3H),: 3.37 (m; 1H), 3.56 (d10; 1H), 3, 64 (m; 1H), 3, 75 (d10; : 1H) and 4.43 (d5; 1H), m / z has 726, 708, 691, 651, 633, 466 448, 354, 314, 297, 265, 247, 219,

and 151.

example

5-Acetoxy and 5,23-Diacetoxy Factor A

Factor A (3.0 g) in pyridine (20 mL) was treated with acetic anhydride (8 mL) at -5 ° and the resulting solution was left at 3 ° for 20 h. After the addition of benzene (100 mL), the solution was concentrated in vacuo. The resulting oil was purified by silica gel column chromatography using dichloromethane: acetone (40: 1) as eluent to give Factor A 5-acetate (2.06 g) containing 5,23-diacetate (10%). Separation of compounds by preparative reverse phase HPLC method, was obtained from 5-acetoxy Factor A (79% yield), X _max (EtOH) 244.5 nm (EJ 462), δ (CDC1 ₃₎ is 2.14 (s, 3H) , m / z has 654, 594 and 576, and 5,23-diacetoxy Factor A (6.5% yield), δ (CDCl ₃ ) has 2.01 (s; 3H) and 2.13 (s; 3H) , m / z has 696 and 636.

example

5,23-Diacetoxy Factor A

A solution of Factor A (600 mg) in purified pyridine (1.0 mL) was treated with excess acid anhydride (0.50 mL) and several crystals of 4-N, N-dimethylaminopyridine. After 24 or. at room temperature, the mixture was poured into ether and then the organic phase was washed successively with 2N chloric acid, saturated sodium bicarbonate solution and, finally, brine. After evaporation of the dry organic phase, the resulting resin was purified using Merck Kieselgei 60, 230-400 silica (45 g) for chromatography. Passing through the column with dichloromethane: ether (9: 1) gave the title compound as a colorless foam (560 mg), [α] 20 D = + 169 ° (c 0.48,

CHCl ₃ ).

Example compound was obtained in a similar manner:

example

5,23-Diacetoxy Factor C

Vol . 211-213 °, [α] g ² + 200 ° (c, 0.40 CHCl ₃ ) , EtOH ' ^A niax 238.5 (30,400 and 245 nm {^ 32,500); _Vmax 3440 (OH) and 1781 cm ¹ (acetoxy and ester) ; δ (CDC1 ₃ ) has 4.90 (m, 1H), 4.04 (d, 6Hz, 1H), 3.96 (d, 10Hz, 1H), 3.58 (m, 1H), 3.32 (m, 1H), 2.14 (s, 3H), 1.75 (s, 3H), 1.67 (d 6Hz, 3H), 1.60 (s, 3H), 1.52 (s, 3H), 0.99 (d, 6Hz, 3H), and 0.71 (d 7Hz, 3H). m / z = = 668 (M +), from Factor C (312 mg). Reaction mixture it is better to pour to ethyl

acetate than into ether and finally crystallized by trituration with diisopropyl ether.

example

23-Acetoxy Factor A

To a stirred and cooled (0-5 °) solution of the compound of Example 6 (530 mg) in methanol (10 mL) was added an aqueous solution of sodium hydroxide (30 mg) (1 mL). After 1.3 or. the mixture was poured into ethyl acetate 3684 B and the organic phase was then washed successively with 2N hydrochloric acid, water, and finally, brine. Evaporation of the dry organic * phase gave a yellow gum which, after being dissolved in dichloromethane, was passed through a Merck Kieselgel 60 column with 230-400 mesh silica (50 g) in the same solvent. Passing through the column with dichloromethane: ether (9: 1) gave the title compound which, upon evaporation of the n-pentane solution, gave an almost colorless solid. This material (330 mg) had [α] ρ ³ + 166 ° (c 0.64 in CHCl 3), λ ^ θ ^Η 239 (26, 500 and 245 nm (8max 29, 300); v _max (CHBr ₃ ) 3540, 3460 (OH) and

1712 -1 cm (ester) ; δ (CDCl ₃ ) has 4.89 (m, 1H), 4.27 (t, 6Hz, 1H), 3.93 (d, 6Hz, 1H), 3.91 (d 10Hz, 1H), 3, 55 (m, 1H), 3.25 (m, 1H), 2.01 (s, 3H), 1.85 (s, 3H), 1.59 (s, 3H), 1.51 (s, 3H), 1.03 (d, 6Hz, 3H), 0.98 (d 6Hz, 3H), 0.94 (d 6Hz, 3H), and 0.69 (d 7Hz, 3H). m / z = 654 (M ⁺ ).

Example compound was prepared in a similar manner to the sample

23-Acetoxy Factor C [α] ρ ³ + 139 ° (c 0.52, CH 2 Cl 2); λ ^ ° ^Η 244.5 nm {8max

29, 400); ? _max (CHBr ₃ ) 3550, 3480 (0H); 1716 {ester and

acetate) and 1255 -1 cm (acetate); δ (CDC1 ₃ ) has 4.91 (q, 3Hz, 1H); 2.03 (s, 3H) and 1.64 (d, 7Hz, 3H) and 1.60 (s, 3H); m / z 626 (M ⁺ ) , from the compound of Example 7 white

in the form of an amorphous solid.

example

5-Acetoxy-23-keto Factor A

A solution of oxalyl chloride (1.96 mL) in dry dichloromethane (25 mL) was treated at -70 ° with a solution of dimethyl sulfoxide (3.19 mL) in dry dichloromethane (15 mL) under nitrogen, followed by dropwise addition of 5-acetoxy A solution of Factor A (4.91 g) in dry dichloromethane (30 mL). The resulting solution was stirred for 1.5 h. At -70 ° before adding a solution of triethylamine (12.6 mL) in dry dichloromethane (40 mL). The reaction mixture was stirred for 1.25 h. without cooling and then added to a cold water / ether (1: 1) mixture. The aqueous layer was extracted with ether. The combined organic layers were washed with water, brine, dried and evaporated. Chromatography of the remaining foam on silica with dichloromethane: acetone (9: 1) gave the title compound (3.4 g), δ (CDCl ₃ ) 3.33 (m; 1H), 3.49 (m, 1H). , 3.70 (d10; 1H) and 5.52 (d5; 1H), m / z has 652, 634,

609, 591, 574, 482, 263, 235 and 151.

example

23-Keto Factor A

To a solution of the compound of Example 10 (276 mg) in methanol (5 mL) at 0 ° was added a solution of 2N-sodium hydroxide (0.42 mL) in methanol (1.0 mL). The solution was left for 5 hours before pouring into cold water. At 5 °. The mixture was extracted with ether and ethyl acetate. The combined organic layers were washed with brine, dried and evaporated to a solid which was purified by preparative thin layer chromatography using dichloromethane: acetone (10: 1) to afford the title compound as a solid.

Compound (140 mg), δ (CDCl ₃ ) 3.28 (m; 1H), 3.48 (m; 1H), 3.70 (d10; 1H), and 4.28 (t R ⁷ ; 1H), m / z 592, 549, 482, 370, 263, 235 and 151.

example

23-Keto Factor A

A spore suspension of 0.4 ml of Streptomyces thermoarchaensis NCIB 12015 dissolved in 10% glycerol was used to inoculate 50 ml of a medium containing 15.0 gL of ¹ D-glucose, 15.0 gL of ' ¹ glycerol, 15.0 g in a 250 ml Erlenmeyer flask. gL ^-1 soy peptone, 3.0 gL ' ¹ NaCl, and 1.0 gL' ¹ CaCO ₃ , and made up to 1 L with additional distilled water prior to autoclaving, the pH was adjusted to 7.0 with aqueous NaOH.

The flask was held for 2 days at 28 ° on a shaker rotating at 250 rpm and the flask moved in a 50 mm diameter circle. Each of the four flat-bottomed 2 L stoppered flasks with 200 ml of the same medium was then inoculated in 4 ml portions, after which the flasks were incubated under the same conditions for 2 days.

The contents of the four flasks were then inoculated with a 70-liter fermenter vessel with the same medium, only supplemented with polypropylene glycol 2000 (0.06%). Propylene Glycol 2000 was added to control foaming during fermentation. Fermentation was carried out at 28 ° with vigorous stirring and aeration to provide dissolved oxygen at levels greater than 30% of saturation. After 24 hours of fermentation, 800 ml and 9 liter portions were transferred to 70 liters of fermentor containing 40 liters of medium and 700 liters of fermentor with 450 liters of medium respectively. The medium in these fermenters was composed of 2.5 gL of ¹ D-glucose; 25.0 gL ¹ malt dextrin (MD30E) 12.5 gL ' ¹ Arkasoy 50; 1.5 gL ' ¹ beet meLT 3684 B lasha; 0,125 GI / ¹ K2HPO4; 1.25 gL ¹ CaCO3 (Ar) and 0.6 gL ' ¹ silicone 1520 (Dow Corning), make up to 1 L with distilled water. Adjust to pH 6.5 with sterile H ₂ SO ₄ before sterilization.

The aforementioned fermentation processes were carried out at 34 ° with vigorous stirring and aeration to provide dissolved oxygen greater than 30% saturation. If necessary, antifoaming polypropylene glycol 2000 was added. After 24 hours. aqueous solution of H ₂ SO ₄ was adjusted to pH 7.2. The fermentation products were harvested and pooled after 4 days.

A micelle (10.4 kg) was collected from the resulting total broth (423 L) by centrifugation in a Sharples PS16AY centrifuge. The micelle was stirred vigorously in methanol (50 L) for 40 min then filtered. The precipitate was resuspended in methanol (27 L) and filtered again. The two filtrates (55 L) were then combined with water (27 L) and 60-80 petroleum ether (30 L) and stirred for 20 min.

The phases were separated in a Westfalia MEM1256 centrifuge, then the methanol phase (75 L) was mixed with additional water (38 L) and 60-80 petroleum ether (30 L). After 20 min, the phases were again separated by centrifugation, and acetone (2 L) was added to the petroleum ether phase to decompose the emulsion. After mixing the methanol phase (110 L) for a third time with water (38 L) and 60-80 petroleum ether (30 L), the phases were separated as before. Again, acetone (3 L) was added to the petroleum ether phase to destroy the emulsion.

The three hexane phases were combined (90 L) and concentrated under low pressure (evaporation temperature 25 °). The concentrate was dried over sodium sulfate (3 Kg) and then evaporated to an oil.

The oil was dissolved in dichloromethane (0.5 L) and filtered through Dicalite 478. The solution (0.9 L) was applied to a column (150 x 10 cm) with silde oxide (Merck) at a rate of 6 L / h, washed with dichloromethane (4 L) and passed through the column with chloroform: ethyl acetate (3: 1 by volume). A fraction washed up between

14.6 and 33.3 L were concentrated to a solid, which in turn was dissolved in a mixture of chloroform: ethyl acetate (3: 1 by volume).

The solution was re-chromatographed on a silica column with the same solvent. The fraction eluted between 14.5 and 31.5 L was dried to a solid and redissolved in chloroform: ethyl acetate (3: 1 by volume). This solution was re-chromatographed on a silica column under the same conditions as before and the fraction eluted between 14 and 31 L was dried to a solid.

The solid was dissolved in 70% v / v aqueous acetonitrile (1.23 L) to which was added the appropriate amount of methanol to make the solution clear. The solution was chromatographed on a 5 ml Spherisorb ODS2 column. 70% acetonitrile was applied to the column at a rate of 20 ml / min, which after 21 min. was increased to 34 mL / min. Fractions of each volume, washed between 12.4 and 16 L, were collected and dissolved in the same volume of water. This solution was then applied to a Montedison S112 column with polystyrene coarse mesh (2 L). The column was washed with 35% acetonitrile and passed through acetone. The fraction washed between 0.5 and 1.25 L was dried to a solid.

The solid was dissolved in acetonitrile (20 mL) and chromatographed on a Sephadex LH20 column in the same solvent. Fractions leaching between 1.08 and 1.26 L were collected and dried to solid state.

The solid was dissolved in 60% acetonitrile (10 mL) with enough ethanol to make a clear solution. It was again chromatographed in 2 ml volumes on a Spherisorb ODS2 column, passing 60% acetonitrile at 25 mL / min. Fractions leached between 0.95 and 1.08 L were collected together and dried to a solid. This solid was dissolved in chloroform (5 mL) and chromatographed on a Merck silica gel 60 column. Subsequently, it was washed with chloroform at 10 mL / min and the fraction eluted between 400 and 790 mL was dried to give the title compound as a solid (33 mg). Nuclear magnetic resonance spectrum confirmed the presence of a 23-keto Factor A.E.I. mass spectroscopic analysis revealed the molecular ion at 610 and the other characteristic fragments were 592, 549, 498, 482, 370, 263 and 151.

Following the application of hplc, it was confirmed that Factor A of Example 23keto is analogous to the ketone produced by the chemical oxidation of Factor A.

example

5-Phenoxyacetoxy-23-deoxy Factor A

To the compound of Example 2 (350 mg) and azobisisobutyrnitrile (25 mg) in dry toluene (20 min) was added a solution of tri-n-butyl tin hydride (0.5 mL) in dry toluene at 120 ° C under nitrogen. In 90 minutes the flocculation solution was cooled and evaporated. Chromatography of the residue on a silica column with dichloromethane: acetone (40: 1) gave

I

38 title compound (280 mg), δ (CDC1 ₃ ) 3, 32 (m; 1H), 3.42 (d10; 1H), 3.57 (m; 1H) , 4.71 (s; 2H), 5, 59 (d6; 1H), and from 6.8 to 7.4 (m; 5H), m / z 7 30 / 712, 578, 560, 468, 450, 356, 314, 299, 249, 248, 221, and 151

example

23-Deoxy Factor A

Example 13 (240 mg) was added to a solution of saturated ammonia in methanol (10 mL) at - 5 °.

The solution was stirred for 2 hours. 0 ° -10 °, then evaporated to dryness. Chromatography of the residue on a silica column using dichloromethane: ace15 (20: 1) as a solvent afforded the title compound (180 mg), δ (CDCl ₃ ) 3.27 (m; 1H), 3.42 (d9; 1H), 3, 54 (m; 1H) and 4.29 (t6; 1H), m / z 596, 578, 560, 468, 450, 356, 314, 299, 249, 248, 221 and 151.

Example 15

5-Acetoxy-23-Mesyloxy Factor A

A solution of 5-acetoxy factor A (3.46 g) in pyridine (30 mL) was cooled in an ice bath and treated with methanesulfonic anhydride (2.2 g). After 30 minutes the solution was left to equilibrate to ambient temperature, then within 60 min. was partitioned between ethyl acetate and hydrochloric acid using successively saturated aqueous sodium bicarbonate solution and saturated saline. After drying the organic solution (Na ₂ SO ₄ ) and evaporating the solvent, the remaining foam was chromatographed on a silica column (Merck Art 9385) with hexane (60 ° -80 °) / ethyl acetate (3: 1) and washed with the same solvent. The combination of the main component of the relevant fractions and evaporation of the solvent resulted in the title compound foam (2.08 g), [α] ^ ² + 154 ° (c = 0.56, CHC1 ₃₎ 247 nm (ε 29,070);

v _max (CHBr ₃ ), 3550, 3470 (OH), and 1735, 1715 (ester); δ (CDCl ₃ ) 4.90 (m, 1H), 3.05 (s, 3H) and 2.16 (s, 3H).

example

23-Tosyloxy Factor B

Factor B (250 mg), tosyl anhydride (204 mg) and several 4-N, N-dimethylaminopyridine crystals were mixed together for 24 h. in dry pyridine (0.5 mL). After pouring the resulting solution into ether, the organic phase was washed successively with 2N hydrochloric acid, saturated aqueous sodium bicarbonate solution and, finally, brine. Evaporation of the dried (Na ₂ SO ₄ ) ether layer afforded a crude material which was loaded with dichloromethane onto a silica gel (50 g, Merck Kieselgel 60, 203-400) column with the same solvent. After washing with dichloromethane: ether (9: 1), the major component was obtained, which was further purified by hplc. Resin-derived titanium

23rd compound [a] p + 173 ° (c , 0.84, CHCl ₃ ); ^{/ v} max (EtOH 235 (30,000) and 244 , 5 nm (s _max 30,800); v _max ( 1708 -1 cm (ester); δ (CDC1 ₃ ) 7.82 (d, 10Hz, 2H), 7.29 (d, 10Hz 2H), 4.83 (kv, 3Hz, 1H), 3.50 (s, 3H) and 2.43 (s, 3H);

m / z = 752 (M +).

example

5-Acetoxy-23-n-butoxy Factor A To a solution of 5-acetoxy Factor A (325 mg) in dry ether was added silver carbonate (1 g) followed by iodobutane (0.5 mL) and silver perchlorate (550 mg). After 20 or so. collidine (0.5 mL) was added to the mixture at room temperature with stirring. After stirring for another 20 min, the mixture was filtered and the filtrate was washed successively with 2N hydrochloric acid, saturated aqueous sodium bicarbonate solution and water. The dried organic phase was evaporated to near dryness and the oil was purified by column chromatography on Merck Kieselgel 60, 203-400. Column using hexane: ethyl acetate (3: 1) as the eluent, there was a colorless foam-like title compound (276 mg) [a] ^ ² + 160 ° (c 0.94, CHC1 ₃₎ X _max (EtOH) 245 nm.

example

Factor A 23-Hemioxalate

Factor A (1 g, about 70% purity) and calcium carbonate (1.5 g) were mixed with dichloromethane (20 mL) at 21 ° and the resulting suspension was added with excess oxalyl chloride (1.0 mL). After 4-5 minutes water (15 mL) was added to the mixture and another 5 min. 2M HCl (10 mL) and ethyl acetate (70 mL). The organic phase was separated, washed with water and brine and then treated with decolorizing charcoal. After 5-10 minutes the organic solution was filtered with phase separation paper and the solvent was separated off from the filtrate by desorption. The precipitate in ether (50 mL) was clarified by filtration through Hyflo and the solvent removed to give a gray-yellow foam. Vigorous shaking with diisopropyl ether (about 7 mL) and washing the resulting solid with diisopropyl ether and n-pentane gave the title compound (750 mg) as a crystalline solid. v _raax (CHBr ₃ ) from 3360 to 3600 (OH), 1805 (acid monomer), and

1720 cm ¹ (acid dimer and esters); δ (CDCl ₃ ) 4.30 (d, 5Hz, 1H) and 5.14 (m, 1H).

example

Factor A

A solution of 23-hemioxalate Factor A (500 mg) in dichloromethane (30 mL) was stirred vigorously with aqueous sodium hydroxide (1.54 g dissolved in 30 mL H ₂ O) for 2 h. At 21 °. After collecting the organic phase, the aqueous phase was washed with a small amount of dichloromethane. The organic phases were combined,

2M HCl was then dried (Na ₂ SO ₄ ) and the title compound was obtained as a foam (430 mg). Based on a comparison made by hplc with an authentic sample of Factor A, the compound was shown to be Factor A.

washed with desorbed sample

Factor A, 5-hemisuccinate

A solution of factor A (306 mg) and succinic anhydride (60 mg) in dry pyridine (0.5 mL) was stirred for 24 h. 22 ° C. After diluting the mixture with ether, the organic phase was washed with 2N hydrochloric acid and brine. After evaporation of the dried ether layer, a foam was obtained from which the title compound was obtained by reverse phase preparative hplc. In ether, the product precipitated out as a white amorphous material with the addition of n-pentane, [α] _D + 125 ° (c 0.44, CHCl ₃ ); ^ ^Η ° λ 245 nm (s _max 30.700) v _max (CHBr ₃ ) 3490 (OH) 3,300-2,00 (CO ₂ H), 1730 (acid carbonyl) and 1710 cm ^-1 (ester); δ (CDCl ₃ ) 2.74 (s,

4H). m / z = 712 (M +).

Example i

5-Acetoxy, 23-Ethyloxyalkyloxy Factor A ·

5-Acetoxy A solution of Factor A (320 mg) and excess ethyl oxalyl chloride in purified dichloromethane (6 mL) was mixed with calcium carbonate (300 mg) for 1 h. The mixture, diluted with ether, was added to saturated sodium bicarbonate and was added for 20 min before being added to ethyl acetate. stirring. Evaporation of the dry (Na ₂ SO ₄ ) organic phase gave a gum (325 mg) which was loaded onto a column of Merck Kieselgel 60, 70-230 silica (25 g) with the same solvent. Rinse the column with dichloromethane and then dichloromethane: ether (95: 5) to give the title compound as a white foam (265 mg) [a] _D + 157 (c)

0.41, CHCl ₃ ); 245 nm (ε _max 32,800); v _max (CHBr ₃ )

3530, 3480 (OH), 1760 and 1735 cm ^-1 (esters); δ (CDCl ₃ ) 5.4 to 5.6 (m, 2H), 5.05 (m; 1H); 4.32 (q, 7Hz,

2H), 2.13 (s, 3H) and 1.35 (t, 7Hz, 3H).

example

23-Methyloxalyloxy Factor A

5-Acetoxy, 23-ethyloxalyloxy A solution of Factor A (50 mg) in methanol (1 mL) containing concentrated sulfuric acid (0.01 mL) was maintained for 17 h. At 21 °. After addition of the mixture to ethyl acetate, the organic phase gradually became neutral. The product was purified by column chromatography on Merck Kieselgel 60, 70-230 silica (25 g), eluting with dichloromethane followed by dichloromethane: ether (9: 1). It was a colorless gum the title compound (25 mg), [ct] p ³ + 147 ° (c 0.28 CHC13) λ ^ ^Η ° 245 nm (sraax 26.600) (ν _max (CHBr ₃ ) 3530, 3480 (OH), 1763, 1737 and 1710 cm ^-1 (esters); δ (CDCl ₃ ) 4.29 (br. vol., 7Hz, 1H), 5.09 (m ,

1H) and 3.88 (s, 3H).

example

5-Acetoxy-23-cyclopropylcarbonyloxy Factor A

5-Acetoxy A solution of Factor A (140 mg) in pyridine (1 mL) was treated with cyclopropanecarboxylic acid chloride (0.08 mL). After 3 or. the reaction mixture was partitioned between ethyl acetate and 2N hydrochloric acid. The organic phase was separated and washed successively with 2N hydrochloric acid and saturated aqueous sodium bicarbonate solution. Drying of the organic solution (Na ₂ SO ₄ ) and evaporation of the solvent gave an oil which was chromatographed on a silica column (Merck Art 9385; 80 mL) with hexane (60 ° 80 °) / ethyl acetate (3: 1) and washed with with the same solvent. Combining the appropriate fractions of the parent compound and evaporating the solvent gave the title compound (70 mg) as a foam. [.alpha.] D @ ₂₀ + 143 DEG (c = 0.54; CHCl3); λ £ $ 5 ¹³ 250 2 (ε = 20310); v _max (CHBr ₃ ) 3550, 3470 (OH) and 1733, 1712 (ester); δ (CDCl ₃ ) 2.16 (s; 3H), 3.33 (m, 1H), 3.88 (d, J 10 Hz, 1H), 5.01 (m; 1H) and (5.5-5 , 6 (m; 2H).

Example compound was obtained in a similar manner:

example

5-Acetoxy-23-cyclopentyloxycarbonyloxy Factor A

5-Acetoxy Factor A (0.28 g) and cyclopentylcarbonylchloride (0.2 mL) gave the foam title compound (0.17 g). [α] D ₂₀ + 149 ° (c = 0.475; CDCl ₃ );

Xmas ^l3 250.2 nm (ε = 19900); ν _max (CHBr ₃ ) 3480 (OH).

example

I

23-Cyclopropylcarbonyloxy Factor A

A stirred solution of 5-acetoxy-23-cyclopropylcarbonyloxy Factor A (230 mg) in methanol (15 mL) was cooled in an ice bath and treated with 3% aqueous sodium hydroxide solution (0.5 mL). After 5 hours, at which time the reaction mixture warmed to room temperature, the solution was partitioned between ethyl acetate and 2N hydrochloric acid. The organic phase was separated and washed successively with 2N hydrochloric acid (x2), dried (Na ₂ SO ₄ ) and the solvent evaporated. The precipitate was chromatographed on a silica column (Merck Art 9385; 150 mL) with hexane (60 ° 80 °) / ethyl acetate (3: 1), and washed with the same solvent. Combining the appropriate fractions of the parent compound and evaporating the solvent gave the title compound as a foam (170 mg), [α] D + 145 ° (c = 0.46, CHCl ₃ ); Xmas ^l3 250.2 nm (s = 20,090); v _max (CHBr ₃ ) 3550, 3480 (OH) and 1709 (esters); δ (CDCl ₃ ) 3.26 (m; 1H), 3.88 (d, J 10Hz, 1H), 4.29 (t, J 7Hz, 1H) and 5.01 (m; 1H).

Example compound was obtained in a similar manner:

example

23-Cyclopentylcarbonyloxy Factor A

Deprotection of 5-acetoxy-23-cyclopentyloxycarbonyloxy Factor A (0.25 g) gave the title compound as a foam (0.215 g) [α] D 20 + 141 ° (c = 0.63; CHCl ₃ ); Ama? ³ 250.2 nm (ε = 19990); v _max (CHBr ₃ ) 3550, 3480 (OH) 1710 (ester); δ (CDCl ₃ ) 2.74 quintet, J 7Hz,

1 H), 3.26 (m; 1H), 3.91 (d, J 10 Hz, 1H), 4.30 (t, J

7 Hz, 1H) and 4.95 (m; 1H).

example

23-Cyclopropylcarbonyloxy-5-methoxycarbonyloxy Factor A

23-Cyclopropylcarbonyloxy A solution of Factor A (0.1 g) in dry dichloroethane (15 mL) was cooled in an ice bath and treated with pyridine (0.2 mL) followed by 1M methyl chloroformate in dichloroethane (0.3 mL). After 20 minutes the solution was diluted with dichloroethane and washed with 2N hydrochloric acid, then dried over sodium sulfate and the solvent evaporated. The precipitate was chromatographed on a silica column (Merck Art 9385; 100 mL) with hexane (60 ° 80 °) / ethyl acetate (3: 1) and washed with the same solvent. Combining the appropriate fractions of the parent compound and evaporating the solvent gave the title compound as a foam (0.106 g). [α] D ₂₀ + 149 ° (C = 0.63, CHCl ₃ ); λ ^ ξ ^ΐ3 3 250.2 (ε = 20470); v _max (CHBr ₃ )

3470 (OH), 1745 (carbonate), 1710 (ester); 998 (CO); δ CDC1 ₃₎ 5.56 (s, 1H), 5.01 (m, 1H), 3.88 (d, J 10Hz,

1 H), 3.83 (s; 3 H), 0.67 (d; J 7 Hz; 1H).

example

Factor A, 5-acetate, 23-isobutyrate

A solution of Factor A, 5-acetate (131 mg), isobutyryl chloride (0.05 mL), pyridine (0.1 mL) and 4-dimethylaminopyridine (25 mg) in dry dichloroethane (10 mL) was stirred for 16 h. At 20 °. The solution was diluted with dichloroethane (30 mL), washed with 2N hydrochloric acid and dried (magnesium sulfate), then evaporated to dryness. The precipitate was chromatographed on Kieselgel 60 (15 g). The column was eluted with petroleum ether: ethyl acetate (4: 1) to give the title compound as a colorless foam (57 mg), [α] θ ° + 159 ° (c = 0.61 in CHCl ₃ ), λ ^ _χ (EtOH) at 244 nm. (ε 30.4000), ^v max (CHBr3) 3480 (OH) and 1713 ^cm-1 (ester), δ (CDC13) 5.5 to 5.6 (m, 2H), 4.95 (m, 1H), 3.93 (d, J 10Hz, 1H), 2.55 (septet, J 7Hz, 1H), 2.17 (s; 3H), 1.20 (d; J 7Hz; 6H) and 0, 70 (d; J 7 Hz; 3H).

example

Factor A, 5,23-di-n-butyrate

A solution of factor A (306 mg), n-butanoic anhydride (0.33 mL) and 4-dimethylaminopyridine (244 mg) in pyridine (5 mL) was stirred for 18 h. At 20 ° C, then added to ethyl acetate and 2N hydrochloric acid (50 mL each). The organic phase was washed with 2N hydrochloric acid, saturated sodium bicarbonate (25 mL) and brine (25 mL), dried (magnesium sulfate) and evaporated to dryness. The precipitate was chromatographed on Kieselgel 60 (40 g). Rinse the column with petroleum ether: ethyl acetate (5: 1) to give the title compound as a colorless foam (200 mg), [α] + 144 ° (c 1.13, CHCl ₃ ), λ _max (EtOH) 245 nm ( ε 31,800), v _max (CHBr ₃ ) 1720 cm ^-1 (est-

ries) , δ (CDC1 ₃₎ from 5, 5 to 5, 6 (m, 2H), 4.94 (m, 1H), 3, 92 (d, J 10Hz, 1H), 2.39 (t, J 7Hz, 2H), 2, 28th (t, J 7Hz, 2H) and 0.70 (d, J 7Hz, 3H), m / z = 752 (M +)

The compounds of Examples 30-32 were obtained in a similar manner:

example

Factor A, 5-acetate, 23-pivaloate (70 mg)

Wd + 159 ° (c 0.69, CHCl ₃ ), (EtOH) 244 nm (ε 29, 900), V _max (CHBr ₃ ) 3470 (OH) and 1730 and 1710 -1 cm (esters), δ (CDC1 ₃ ) from 5.5 to 5.5, m (2H), 4, 95 (m, 1H), 3.91 (d, J 10Hz, 1H) , 2.16 (s, 3H), 1.22 (s, 9H), and 0, 68 {d, J

7Hz, 3H), from Factor A, 5-acetate (131 mg) and pivaloyl chloride (0.06 mL).

example

Factor A, 5-acetate, 23-benzoate (180 mg) [α] D ₂₀ + 153 ° (c 0.59 in CHCl ₃ ), (EtOH) 236 nm (ε 36,200), ν _max (CHBr ₃ ) 3460 (OH). ) and 1730 and 1707 cm ^-1 (esters),

δ (CDCl ₃ ) 8.21 (d, J 7Hz, 2H), 7.56 (t, J 7Hz, 1H), 7.44 (t, J 7Hz, 2H) from 5, 5 to 5.6 (m, 2H), 4, 07 (d, J 10Hz, 1H), 2.17 (s, 3H) and 0.75 (d, J 7Hz, 3H), of Factor A, 5-acetate (327 mg) and benzoines of acid

anhydride (339 mg).

example

Factor A, 5,23-dibenzoate (370 mg) and 5-benzoate (200 mg)

From Factor A (613 mg) and benzoyl chloride (0.35 mL); from the initial fractions, the titled dibenzoate was obtained, [α] D + 86 ° (c 0.65, CHCl ₃ ), (EtOH) 236 nm (ε 46,200), ν _max (CHBr ₃ ) 3480 (OH), 1710 (esters). ), and 1602 and 1585 cm ^-1 (phenyl), δ (CDCl ₃ ) 8.2 to 8.0 (m, 4H), 7.7 to 7.5 (m, 2H), 7.46 (t , J 7Hz, 4H), 5.61 (s, 1H), 4.07 (d, J 10Hz, 1H), and 0.76 (d, J 7Hz, 3H).

The following fractions were obtained main benzoate [a] § ° + 80 ° (c 0.61, CHC1 ₃₎ \ _nax (EtOH) 237 nm (ε 39.200), ^v max (CHBr ₃₎ 3500 (OH), 1712 (esters), and 1601 and

1585 cm ^-1 (phenyl), δ (CDCl ₃ ) 8.09 (d, J 7Hz, 2H), 7.58 (t, J 7Hz, 1H), 7.45 (t, J 7Hz, 2H), δ , 60 (s, 1H), 3.83 (m, 1H), 3.76 (d, J10H2, 1H), and 0.81 (d, J7Hz, 3H).

example

Factor A, 5-chloroacetate

A solution of Factor A (123 mg) and pyridine (0.1 mL) in dry dichloromethane (5 mL) was stirred at 0 ° and chloroacetyl chloride (0.3 mL of 1M solution in dichloromethane) was added. The resulting solution was stirred for 15 min, then chloroacetyl chloride (0.1 mL of 1M solution) was added and stirred for a further 15 min. The solution was diluted with dichloromethane (50 mL), washed with 2N hydrochloric acid (2 x 20 mL) and saturated sodium bicarbonate (20 mL), dried (magnesium sulfate) and evaporated to dryness. The residue (140 mg) was purified by chromatography on Kieselgel 60 (15 g). The column was washed with petroleum ether / ethyl acetate (4: 1) to give the title compound as a colorless foam (90 mg), [α] D + 143 ° (c 1.11, CHCl ₃ ), (EtOH) 245.5 nm (ε 30,400). ), v _max (CHBr ₃ ) 3500 (OH), 1760 (chloroacetate), and 1710 cm ^-1 (ester), δ (CDCl ₃ ) 5.5 to 5.6 (m, 2H), 4.18 (s) , 2H), about 3.81 (m, 1H), 3.74 (d, J 10Hz, 1H) and 0.80 (d, J 7Hz, 3H), m / z = 752 (M +, ³⁵ Cl).

Examples 35, 45, 49 and 50 were obtained in a similar manner.

example

Factor A, 5-chloroacetate, 23-ketone

A solution of Factor A, 5-chloroacetate (276 mg) and pyridine dichromate (602 mg) in dry N, N-dimethylformamide (10 mL) was stirred for 48 h. At 20 ° C, then added to a mixture of ethyl acetate (50 mL) and 2N hydrochloric acid (25 mL). The organic phase was washed with 2N hydrochloric acid and saturated sodium bicarbonate, dried (magnesium sulfate) and evaporated to dryness. The precipitate was purified by chromatography on Kieselgel 60 (25 g). The column was washed with petroleum ether: ethyl acetate (4: 1) to give the title compound as a colorless foam (85 mg), [α] D ₂₀ + 120 ° (c 1.05 in CHCl ₃ ), (EtOH) at 245 nm (ε 26,500). , ^v max (CHBr 3) 3480 (OH), 1760 (chloroacetate) and 1715 cm ^-1 (ester and ketone), δ (CDCl 3) 5.5 to 5.7 (m, 2H), 4.17 (s, 2H), 3.72 (d, J 10 Hz, 1H), 2.50 (s, 2H) and 0.85 (d, J 7 Hz, 3H), m / z = 686 (M +, ³⁵ Cl).

example

Factor A, 5-bromoacetate (447 mg) [α] D ₂₀ + 132 ° (c 0.77, CHCl ₃ ), (EtOH) 245.5 nm (ε 32,500), ν _max (CHBr ₃ ) 3495 (OH). , 1730 (bromoacetate) and 1712 cm ^-1 (ester), δ (CDCl ₃ ) 5.5 to 5.6 (m, 2H), 3.97 (d, J 12 Hz, 1H), 3.90 (d) , J 12Hz, 1H), 3.81 (m, 1H), 3.75 (d, J 10Hz, 1H) and 0.81 (d, J 7Hz, 3H) from Factor A (613 g) and bromoacetyl bromide (0.13 mL).

example

Factor A, 5,23-chloroacetate

A solution of Factor A, 5-chloroacetate (365 mg), chloroacetyl chloride (0.21 mL) and calcium carbonate (265 mg) in dry dichloromethane (15 mL) was stirred for 48 h. At 20 °. The resulting suspension was dissolved in dichloromethane (50 mL), then washed with 2N hydrochloric acid and saturated sodium bicarbonate, dried (magnesium sulfate) and evaporated to dryness. The precipitate was purified by chromatography on Kieselgel 60 (30 g). The column was washed with petroleum ether: ethyl acetate (4: 1) to give the title compound as a colorless foam (167 mg), [α] D ₂₀ + 151 ° (c 1.17, CHCl ₃ ).

Λ _max (EtOH) 245, 5 nm (ε 29,400), ν _max (CHBr ₃ ) 3470 (OH), and 1725 cm ^-1 (esters), δ (CDCl ₃ ) 5.5 to 5.6 (m, 2H). ),

5.00 (m, 1H), 4.17 (s, 2H), 4.10 (d, J 15Hz, 1H), 4.02 (d, J 15Hz, 1H), 3.92 (d, J 10Hz) , 1H) and 0.73 (d, J *

7Hz, 3H), m / z = 764 (M +, ³⁵ Cl).

Examples 37, 38 and 46 were obtained in a similar manner:

example

Factor A, 23-methoxyacetate (92 mg)

[α] ²⁰ D + 157 ° (c, 0.75, CHCl ₃ ), λ ^ _χ (EtOH) 244 nm (ε 32,300), v _max (CHBr ₃ ) 3560 and 3480 (OH), 1740 (methoxyacetate) and 1712 cm ¹ (ester), δ (CDC1 ₃₎ 5, 01 (m, 1H), 4.28 (t, J 7 Hz, 1H), 4.01 (s, 2H) r, 3.89 (d, J 10 Hz, 1H), 3.46 (s, 3H) and 0.71 (d, J 7Hz,  3H), from Factor A (184 mg)

and methoxyacetyl chloride (0.3 mL).

example

Factor A, 23-phenoxyacetate (112 mg) [α] g ° + 129 ° (c 0.75, CHCl ₃ ), (EtOH) 244 nm (ε 25.900), v _max (CHBr ₃ ) 3550 and 3470 (OH). , and 1745 and 1708 cm ^-1 (esters), δ (CDCl ₃ ) 7.30 (t, J 8Hz, 2H), 7.00 (t, J

8Hz, 1H), 6.94 (d, J8Hz, 2H), 5.02 (m, 1H), 4.68 (d, J16Hz, 1H), 4.58 (d, J16Hz, 1H), 4.30 J 7Hz, 1H), 3.92 (d, J 10Hz, 1H) and 0.67 (d, 7Hz, 3H) from Factor A (184 mg) and phenoxyacetyl chloride (0.38 mL).

example

Factor A, 5,23-di- (2,2,2-trichloroethyl) carbonate (362 mg) [α] D ₂₀ + 110 ° (c 0.94, CHCl ₃ ), (EtOH) 244 nm (ε 28,000) , v _max (CHBr ₃ ) 3550 and 3460 (OH), 1750 (carbonate) and

1710 cm- ¹ (ester), δ (CDCl ₃ ) 5.60 (s, 1H), 4.89 and 4.75 (AB sq, J 12Hz, 2H), 4.85 (m, 1H), 4.81 and 4.71 (AB sq. J 14Hz, 2H) and 0.79 (d, J 7Hz, 3H), from Factor A (306 mg) and 2,2,2-trichloroethyl chloroformate (0.14 mL).

example

Factor A, 23- (2,2,2-trichloroethyl) carbonate

A solution of Factor A, 5,23-di- (2,2,2-trichloroethyl) carbonate (200 mg) in dioxane (10 mL) was stirred at about 20 ° and 1M-sodium hydroxide (0.5 mL) was added. The resulting solution was stirred for an additional 30 min, then 1M sodium hydroxide (1 mL) was added and stirred for another 1 h. The solution was diluted with ethyl acetate (50 mL), washed with 2N hydrochloric acid, dried (magnesium sulfate) and evaporated to dryness. The residue was purified by chromatography on Kieselgel 60 (15 g). Rinse the column with petroleum ether: ethyl acetate (2: 1) to give the title compound as a colorless foam (80 mg), [α] θ ° + 142 ° (c 0.92, CHCl ₃ ), λ _max (EtOH) 245 nm ( ε 29,200), v _max (CHBr ₃ ) 3560 and 3500 (OH), 1745 (carbonate) and 1710 cm ^-1 (ester), δ (CDCl ₃ ) 5.40 (s, 1H),

4.80 and 4.70 (AB kv, J 14Hz, 2H), 4.28 (m, 1H) and 0.79 (d, J 7Hz, 3H).

example

Factor A, 5-acetate, 23- (2,2,2-trichloroethyl) carbonate (385 mg) [α] D ₂₀ + 140 ° (c 1.07, CHCl ₃ ), λ _max (EtOH)

245, 5 nm (ε 29,200), v _max (CHBr ₃ ) 3300 to 3620 (OH), 1738 (carbonate) and 1720 cm ^-1 (ester), δ (CDCl ₃ ) has 5.53 (s, 1H). , 4.81 and 4.70 (AB kv, J 14Hz, 2H), 4.84 (m, 1H), 3.98 (d, J 10Hz, 1H), 2.16 (s, 3H) and 0, 79 (d, J 7 Hz, 3H) from Factor A, 5-acetate (393 mg) and

2,2,2-trichloroethyl chloroformate (2 mL of a 1M solution in dichloromethane).

example

Factor A, 5,23-di-n-butyrate

A solution of factor A (306 mg), n-butanoic anhydride (0.33 mL) and 4-dimethylaminopyridine (244 mg) in pyridine (5 mL) was stirred for 18 h. At 20 ° and then added to a mixture of ethyl acetate and 2N hydrochloric acid (50 ml each). The organic phase was washed with 2N hydrochloric acid, saturated sodium bicarbonate (25 mL) and brine (25 mL), dried (magnesium sulfate) and evaporated to dryness. The precipitate was purified by chromatography on Kieselgel 60 (40 g). The column was washed with petroleum ether: ethyl acetate (5: 1) to give the title compound as a colorless foam (200 mg), [α] D + 144 ° (c 1.13, CHCl ₃ ), (EtOH) 245 nm (ε 31.800). , (CHBr ₃ )

1720 cm ^-1 (ester), δ (CDCl ₃ ) 5.5 to 5.6 (m, 2H),

4.94 (m, 1H), 3.92 (d, J 10Hz, 1H), 2, 39 (t, J 7Hz, 2H), 2.28 (t, 752 (M +). J 7Hz 2H) and 0.70 (d, J 7Hz, 3H), m / z = Example 43 Factor A, 23-n-Butyrate Factor A 5,23-di-n-butyrate (160 mg) in methanol

(10 mL) was stirred at 0 ° C and 1M-sodium hydroxide (0.25 mL) was added. The resulting solution was stirred for a further 90 min. at 0 to 5 °, then ΙΜ-sodium hydroxide (0.25 mL) was added and stirred for a further 4 h. The solution was added to a mixture of ethyl acetate (50 mL) and 2N hydrochloric acid (25 mL), then the organic phase was washed with water, brine and dried (magnesium sulfate) and evaporated to dryness. The precipitate was purified by chromatography on Kieselgel 60 (20 g). After washing the column with light petroleum: ethyl acetate (2: 1) was obtained a colorless foam-like title compound (68 mg), [a] p ° + 164 ° (c 1.03, CHC1 ₃₎ l _max (EtOH) 244.5 nm (ε 29, 600), ν _max (CHBr ₃ ) 3550 and 3470 (OH), and 1710 cm ^-1 (esters), δ (CDCl ₃ ) 4.93 (m, 1H),

4.28 (t, J 7Hz 1H), 3.91 (d, J 10Hz, 1H), 2.28 (t, J 7Hz, 2H), 1.65 (m, 2H), 0.94 (t, J 7 Hz, 3 H) and 0.69 (d, J 7 Hz, 3 H), m / z = 682 (M +).

example

Factor A, 23-methyl carbonate (155 mg) [α] ρ ° + 169 ° (c 0.81, CHCl ₃ ), λ ^ _χ (EtOH) 245 nm (ε 26,600), v _max (CHBr ₃ ) from 3300 to 3610 (OH), 1735 (carbonate) and 1710 cm ^-1 (ester), δ (CDCl ₃ ) 5.40 (s, 1H), 4.28 (m,

1H), 3.94 (d, J 10Hz, 1H), 3.77 (s, 3H), and 0.76 (d, J

7Hz, 3H), m / z = 670 (M +), from Factor A, 5-acetate, 23 (2,2,2-trichloroethyl) carbonate (300 mg).

example

23-Deoxy Factor A, 5-methylcarbonate (57 mg) [α] θ ° + 152 ° (c, 0.6, CHCl ₃ ), λ _max (EtOH) 244.5 nm (ε 28,200), ν _max (CHBr) ₃₎ 3530 and 3460 (OH), 1740 (carbonate) and 1707 ^cm-1 (ester), δ (CDC1 ₃₎ 5.55 (s, 1H)

3.82 (s, 3H), 3.42 (d, J 10Hz, 1H) and 0.69 (d, J 4Hz, 3H), from 23-deoxy Factor A (90 mg) and methyl chloroformate (0.3 mL) Of ΙΜ-solution in dichloromethane).

example

Factor A, 23-chloroacetate (193 mg) [α] g ° + 162 ° (c 1.04, (CHCl ₃ ), λ _max (EtOH) 245 nm (ε 28,900), ν _max (CHBr ₃ ) from 3320 to 3620 (OH), 1748 (chloroacetate) and 1710 ^cm-1 (ester), δ (CDC1 ₃₎ is 5.42 (s, 1H), 4.28 (m, 1H), 4.09 and 4.01 (AB sq, J 15Hz, 2H), 3.91 (d, J 10Hz, 1H) and 0.73 (d, J 7Hz, 3H), m / z = 688 (M +, ³⁵ Cl), from Factor A (306 mg). ) and chloroacetyl chloride (0.2 mL).

example

Factor A, 5,23-dipropionate (387 mg) [α] θ ° + 157 ° (c, 0.96, CHCl ₃ ), k _max (EtOH) 244.5 nm (ε 30,200), v _max (CHBr ₃₎ ) 3500 (OH) and 1720 cm ^-1 (esters), δ (CDCl ₃ ) has 5.52 (s, 1H), 3.93 (d, J 10Hz, 1H), 2.43 (sq, J 7Hz, 2H), 2.32 (sq, J 7Hz, 3H), 1.18 (t, J 7Hz, 3H), 1.16 (t, J 7Hz, 3H) and 0.70 (d, J 7Hz, 3H) , m / z = 724 (M +), from Factor A (613 mg) and propanoic anhydride (0.5 mL).

example

Factor A, 23-propionate (155 mg) [α] β ° + 168 ° (c 1.03, CHCl ₃ ), (EtOH) 244.5 nm (ε 30, 600), v _max (CHBr ₃ ) 3550 and 3480 (OH), and 1710 cm ^-1 (ester), δ (CDCl ₃ ) 5.39 (s, 1H), 4.27 (m, 1H), 3.91 (d, J 10 Hz, 1H), 2, 31 (sq, J 7Hz, 2H), 1.14 (t, J 7Hz, 3H) and 0.69 (d, J 7Hz, 3H), m / z = 668 (M +), from Factor A, 5.23 -dipropionate (327 mg) in a similar manner to the compound of Example 43.

example

Factor A, 5-methyl carbonate, 23-ketone (830 mg) [α] D ₂₀ + 132 ° (c 0.82, CHCl ₃ ), k _max (EtOH), 245 nm (ε 29.800). Infrared and nuclear magnetic resonance spectroscopy data were similar to that of the compound described in Example 40 from Factor A, 23-ketone (916 mg) and methyl chloroformate (0.23 mL).

example

Factor A, 5-benzyl carbonate, 23-ketone (57 mg) [α] ρ ° + 99 ° (c 0.4, CHCl ₃ ), (EtOH) 245.5 nm (ε 29, 600), ^v max ( CHBr ₃ ) 3600, 3550 and 3480 (OH), 1740 (carbonate)

and 1715 cm ¹ (ester and ketone), δ (CDC1 ₃₎ from 7.5 to 7.2 (m, 5H), 5.57 (s, 1H), 5.21 (s, 2H), 3, 70 (d, J 10Hz, 1H), 2.49 (s, 2H) and 0.86 (d, J 6Hz, 3H), of

Factor A, 23-ketone (92 mg) and benzyl chloroformate (0.05 mL).

example

5-Acetoxy-23-n-butoxy Factor A

To a solution of 5-acetoxy Factor A (325 mg) in purified ether was added silver carbonate (1 g) followed by iodobutane (0.5 mL) and silver perchlorate (550 mg). The solution was stirred for 20 hours. at room temperature, then collidine (0.5 mL) was added. After stirring for another 20 min, the mixture was filtered and the filtrate was washed successively with 2N hydrochloric acid, saturated aqueous sodium bicarbonate solution and water. The dried organic phase was evaporated to near dryness and the oil was purified by chromatography on Merck Kieselgel 60 with 230-400 silica (100 mL). After washing the column with hexane: ethyl acetate (3: 1) was obtained a colorless foam-like title compound (276 mg, 78%) [(x] q ² + 160 ° (c 0.94,

CHCl3), X _max (EtOH) 245 nm (ε 28.300); δ (CDCl ₃ ) 3.16 (m, 1H), 3.43 (m, 1H), 3.59 (m, 1H).

example

5-Acetoxy-23-cyclopropylmethoxy Factor A [α] at ¹ + 174 ° (c 1.74, CHCl ₃ ), k _max (EtOH) 244 nm (s _max

28,360), v _max (CHBr ₃ ) 3470 (OH), and 1732, 1710 (esters), and 998 cm ^-1 (CO) δ (CDCl ₃ ) have 5.5-5.6 (m; 2H), 3.96. (d 10; 1H), 3.49 (m; 1H), 3.37 (dd 6, 10; 1H), 3.17 (dd, 6.10; 1H), 2.16 (s; 3H), 0.76 (d 7; 3H), 0.44 (m; 2H), 0.22 (m; 2H) from 5-acetoxy Factor A and bromomethylcyclopropane, similar to Example 17.

example

5-Butyldimethylsilyloxy-23-methoxy Factor A

A solution of methylmagnesium iodide in ether (0.35 mL of a 3M solution) was added at room temperature under nitrogen atmosphere to a solution of 5-t-butyldimethylsilyloxy-Factor A (239 mg) in dry hexamethylphosphoric triamide (18 mL) stirred under a magnetic stirrer. After 30 minutes after addition of iodomethane (0.4 mL), the resulting mixture was stirred for 5 h, diluted with ether (100 mL) and rinsed well with water. The ether solution was then washed with brine (50 mL) and the organic phase was evaporated. The resulting foam (280 mg) was purified by chromatography on a 230-400 mesh Merck Kieselgel 60 column (80 mL). After washing the column with hexane: ethyl acetate (5: 1) was obtained a colorless foam-like title compound (46%) [a] ^ ¹ + 142 ° (c 1.13, CHC1 ₃₎ (EtOH) 244 nm (ε 27 900);

^v max (CHBr3) 3460 (OH), 1708 (broad; ester) and 995 cm @ ^-1 (CO); δ (CDCl 3) 4.42 (m; 1H), 3.92 (d 10; 1H), 3.3-3.4 (m; 2H), 3.32 (s; 3H), 0.92 (s) ; 9H), 0.75 (d 7; 3H), 0.12 (s; 6H).

Example compound was obtained in a similar manner:

example

5-t-Butyldimethylsilyloxy-23-n-propyloxy Factor A [α] + 153 ° (c 1.16, CHCl ₃ ), (EtOH) 244 nm (ε ^ 29.950),? _Max (CHBr ₃ ) 3460 (OH ), 1705 (ester) and 995 cm ^-1 (CO), δ (CDCl ₃ ) 4.44 (m, 1H), 3.96 (d 10; 1H), 3.56 (m; 1H),

3.45 (m; 1H); 3.14 (m; 1H); 0.93 (s, 9H); 0.75 (d, 7; 3H); 0.13 (s, 6H); butyldimethylsilyloxy Factor A and 1-iodopropane.

example

5-Acetoxy Factor D (923 mg) [α] D ²⁰ + 143.9 ° (c 0.9, CHCl ₃ ), λ ^ $ ^Η 239 (28,700) and 245 nm (s _max 31,000); v _max (CHBr ₃ ) 3490 (OH), 1730 and 1710 (ester); Δ (CDCl ₃ ) 0.81 (d, 7Hz; 3H), 0.99 (d, 7Hz,

3H), 1.00 (t, 7Hz, 3H), 1.53 (s; 3H), 1.59 (s; 3H),

1.75 (s; 3H), 2.16 (s; 3H), 3.32 (m; 1H), 3.65 (m; 1H),

4.04 (d, 6Hz; 1H) and 5.35 (m; 2H), m / z = 640 (M +),

Factor D (2.5 g) and acetic anhydride (0.47 mL) were similar to the compound of Example 5.

example

5,23-Diacetoxy Factor D (286 mg)

EurLex-2 en 147-149 ° diamonds of pentane ether [α] D + 152.6 ° (c 0.9, CHCl ₃ ) x x _a 5 ^H 232.5 (21,500), 238 (26,600), and

244.5 nm (δ _max 28,800); v _max (CHBr ₃ ) 1720 cm ^-1 (ester); δ (CDCl ₃ ) 0.72 (d, 6Hz; 3H), 0.99 (d, 6Hz; 3H), 1.01 (t, 7Hz; 3H), 1.53 (s, 3H), 1.60 (s; 3H), 1.75 (s; 3H),

2.02 (s; 3H), 2.15 (s; 3H), 3.31 (m; 1H), 4.04 (d, 6Hz, 1H), 4.91 (m; 1H) and from 5.5 to 5.6 (m; 2H). m / z = 682 (M +), from Factor D (439 mg) and acetic acid anLT 3684 B hydride (0.25 mL), similar to that of Example 6.

example

23-Acetoxy Factor D (127 mg)

MS ¹ (^ max + 150 ° (c 0.5, CHC1 ₃₎ (CHBr ₃₎ ^H max ^X 238 (29.200), and 244.5 nm 15,600); v ^y max 3300, 3590 (OH) and 1710 cm ^-1 (ester); δ (CDCl 3) 0.72 (d, 7Hz; 3H), 0.99 (d, 7Hz; 3H), 1.01 (t, 7Hz ; 3H), 1.53 (s, 3H), 1.60 (s, 3H), 1.86 (s, 3H), 3.26 (m, 1H) , 3.95 (d, 6Hz; 1H), 4.26 (t, 6Hz; 1H), and 4, 91 (m, 1H). m / z = 640 (M +), of 5.23-

diacetoxy Factor D (207 mg), similar to the compound of Example 8.

example

5-Acetoxy, 23-keto Factor D (152 mg)

lt 228-230 ° [a] ^ ¹ + 84 ° (c 0.6, CHC1 ₃₎ 244.5 nm _(max £ 31.100) v _max (CHBr ₃ ) 3500 (OH), 1732 and 1714 cm ¹ (ester and ketone); δ (CDCl ₃ ) 0.86 (d, 6Hz; 3H), 0.98 (d, 7Hz; 3H), 1.00 (t, 7Hz; 3H), 1.49 (s, 3H), 1.67 (s; 3H), 1.74 (s; 3H), 2.14 (s; 3H), 3.33 (m; 1H), 4.03 (d,

6Hz; 1H) and 5.5-5.6 (m; 2H). m / z = 638 (M +). From 5acetoxy Factor D (336 mg), similar to the compound from Example 10.

example

23-Keto Factor D (59 mg) [ajg ¹ + 84 (c 0.4, CHCl ₃ ), 244, 5 nm (s _max 28,000);

v _max (CHBr ₃ ) 3550 and 3500 (OH), and 1712 cm ¹ (ester and ketone); δ (CDCl ₃ ) 0.86 (d, 6Hz; 3H), 0.98 (d, 7Hz;

3H), 1.00 (t, 7Hz; 3H), 1.50 (s, 3H), 1.68 (s, 3H),

1.86 (s, 3H), 3.27 (m, 1H), 3.73 (d, 10 Hz, 1H), 3.95 (d, 6 Hz, 1H), and 4.27 (t, 6 Hz) ; 1H), from 5-acetoxy-23keto Factor D (96 mg), similar to the compound of Example 11.

example

Factor A, 23-phenylacetate (240 mg) [α] θ ° + 140 ° (c, 0.92, CHCl ₃ ), v _raax (CHBr ₃ ) 3550 and 3470 (OH), and 1730 cm ¹ (esters), δ (CDCl ₃ ) 7.32 (s, 5H),

5.0m, 1H), 4.29 (t, J7Hz, 1H), 3.88 (d, J10Hz, 1H), 3.62 (s, 2H) and 0.54 (d, J7Hz), 3H) was obtained in a similar manner to the compound of Example 38 from Factor A (306 mg) and phenylacetate chloride (0.33 mL).

example

23-Ethoxy Factor A

5-Acetoxy-23-ethoxy A solution of Factor A (806 mg) in methanol (18 mL) was cooled in an ice bath, 1N aqueous sodium hydroxide (1.3 mL) was added and the resulting light yellow solution was stirred for 1.25 h. The solution was diluted with ethyl acetate (80 mL), then washed successively with IN hydrochloric acid, water and brine. The dry organic phase was evaporated and the resulting resin was purified by chromatography on a 230-400 mesh Merck Kieselgel 60 column (200 mL). The column was eluted with 15% ethyl acetate in dichloromethane to give the title compound as a colorless foam (623 mg) [α] 20 D + 178 ° (c 1.13,

CHC1 _3), X _max (EtOH) 244 nm (ε 29 400); δ (CDCl ₃ ) 4.29 (t, 7; 1H), 3.65 (m; 1H), 3.47 (m; 1H), 3.26 (m; 2H), 1.15 (t, 7 ; 3H).

Examples 123-126 were obtained in a similar manner:

example

23-n-butoxy Factor A (61%) were obtained as a colorless foam [a] ^ ¹ + 161 ° (c 1.47, CHC1 _{3). Λmax} (EtOH) 244 nm (ε 33, 100); δ (CDCl ₃ ) 4.30 (t, 7; 1H), 3.60 (m; 1H),

3.43 (m; 1H), 3.17 (m; 2H) from 5-acetoxy-23-n-butoxyFactor A.

example

23-n-Propoxy Factor A (83%) was obtained as a colorless foam [a] g ¹ + 165 ° (c 1.01, CHCl ₃ ), λ _max (EtOH) 245 nm (δ 30, 970); δ (CDCl ₃ ) 4.29 (t, 7; 1H), 3.55 (m; 1H),

3.44 (m; 1H), 3.13 (m; 1H) from 5-acetoxy-23-n-propoxyFactor A.

example

23-Methoxy Factor A (66%) was obtained as a colorless foam [ccjp ¹ + 175 ° (c 1.01, CHCl ₂ ), k _max (EtOH) at 244 nm (ε

19,100); δ (CDCl ₃ ) 4.29 (t, 7; 1H), 3.40 (m; 1H), 3.33 (s; 3H), from 5-acetoxy-23-methoxy Factor A.

example

23-Cyclopentyloxy Factor A (75%) was obtained as a colorless foam [α] θ + + 160 ° (c 1.65, CHCl ₃ ), k _max (EtOH)

244 nm (ε 19, 800); δ (CDCl ₃ ) 4.29 (t, 7; 1H), about 3.96 (weak m; 1H), 3.95 (d, 5; 1H), 3.91 (d, 10; 1H),

3.46 (m; 1H), 0.69 (d, 7; 3H) from 5-acetoxy-23-cyclopentyloxy Factor A.

Example (a) 5-Acetoxy-23-allyloxy Factor A

Silver salicylate (872 mg) was added to a solution of 5-acetoxy Factor A (207 mg) and allyl iodide (1.0 mL) in dry ether (25 mL) and the mixture was stirred for 4 days at room temperature before filtration. Evaporation of the filtrate gave a yellow oil which was purified by chromatography on a 230-400 mesh Merck Kieselgel 60 column. After washing with dichloromethane: ethyl acetate (19: 1) was obtained a colorless foam-like title compound (105 mg), [<x] q + 152 ° (c 1.00 CHC1 ₃₎ i _max (EtOH) 245 nm (ε 28,400 , δ (CDCl ₃ ) 3.54 (m, 1H), 4.14 (m, 1H).

The following compounds were obtained in a similar manner:

(b) 5-Acetoxy-23-n-propyloxy Factor A, from 5-Acetoxy Factor A and n-propyl iodide. After purified by chromatography on 230-400 mesh, Merck Kieselgel 60, elution column using hexane: ethyl acetate (3: 1) of 22 as a colorless foam-like title compound [a] _D + 160 ° (c 0.75 CHC1 _3), X _max ( EtOH) 245 nm (ε, 27,600), δ (CDC1 ₃₎ 3.33 (s, 3H), 3.39 (m, 1H).

(c) 5-Acetoxy-23-methoxy Factor A, from 5-acetoxy

Factor A and methyl iodide. The title compound was obtained in the form of a colorless 9 9 foam [a] _D ⁺ 159 ° (c 0.98).

CHCl ₃ ), _λmaK (EtOH) 245 nm (ε 27600), δ (CDCl ₃ ) 3.33 (s,

3H), 3.39 (m, 1H).

example

5-Acetoxy-23-ethoxy Factor A from 5-Acetoxy Factor A and ethyl iodide. The title compound was obtained in the form of a colorless foam, [α] 20 D + 167 ° (c 1.02 CHCl ₃ ), k _max (EtOH) 245 nm (ε 28070), δ (CDCl ₃ ) 3.25 (m, 1H),

3.46 (m, 1H), 3.64 (m, 1H), in a similar manner to Example 17.

Claims (8)

DEFINITION OF INVENTION wherein R ¹ represents a methyl, ethyl or isopropyl group; R represents a hydrogen atom or an OR group (where OR is a hydroxyl group or a substituted hydroxyl group having up to 25 carbon atoms) and R ³ represents a hydrogen atom, or 2 3 R and R together with the carbon atom to which they are attached represent a) C = O; and 4 5 OR denotes OR as defined above; provided that when R ² represents a hydroxyl group, OR ⁴ is a substituted hydroxyl group but not a methoxyl group. 2. A compound according to claim 1, characterized in that R ¹ is an isopropyl group. 3. A compound according to claim 1 wherein R is isopropyl, R is a hydrogen atom or pionoxyl group of ethoxyl, n-propoxyl, acetoxyl or proLT 3684 B, and R is a hydrogen atom or R and R ³ together with a carbon atom, to which they are attached form) C = O, and OR ⁴ is a hydroxyl, acetoxyl or methyloxycarbonyloxy group. 4. A compound according to claim 1 wherein: R ¹ is an isopropyl group, R ² is a hydrogen atom, R ³ is a hydrogen atom, and OR ⁴ is a hydroxyl group; R is an isopropyl group, R is a propionoxyl group, R is a hydrogen atom, and OR ⁴ is a hydroxyl group; R ¹ is present isopropyl group R ² and R ³ together with carbon atomic, to which they are online, indicates > C - 0 group and -OR ⁴ is a hydroxyl group; R ¹ is present isopropyl 2 3 group, R is ethoxyl group, R is the hydrogen atom and -OR ⁴ are a hydroxyl group; R ¹ is isopropyl group, R ² is n-propoxyl group, R ³ is hydrogen atom and -OR ⁴ hydroxyl group; R ¹ is a methyl group, R ² is an acetoxyl group, R ³ is a hydrogen atom and OR ⁴ is a hydrogen atom; R is an ethyl group, R is an acetoxyl group, R is a hydrogen atom and OR ⁴ is a hydroxyl group; or 1 2. . R 3 is an isopropyl group, R is an acetoxyl group, R is a hydrogen atom, and OR ⁴ is a hydroxyl group. 5. A composition for human and veterinary use, which comprises, together with one or more carriers and / or excipients, an effective amount of at least one compound according to claim 1. A pest control composition for controlling pests, for example, in agriculture, horticulture or forestry, which comprises, together with one or more carriers and / or excipients, an effective amount of at least one compound according to claim 1. 7. A process for the preparation of a compound according to claim 1, wherein: (a) in which one or both of R and OR is a substituted hydroxyl group which is obtained by interaction ati2 4 suitable compound in which one or both R and OR is a hydroxyl group with a hydroxyl shifting reagent interaction OR ^{2 * 4} in protected hydroxy to give compounds wherein OR is -OH; 2 3 (b) a compound wherein R and R are both hydrogen atoms obtained by reduction of a corresponding compound wherein R is a homolytically reducing atom or group; 2 3 (c) a compound wherein R and R together represent an oxo group obtained by oxidation of the corresponding compound wherein R is -OH, OR ⁴ is a protected hydroxyl group to give compounds wherein OR ⁴ is -OH; (d) a compound wherein R ² or OR ⁴ is a hydroxyl group, by deprotecting the corresponding compound wherein R or OR is a protected hydroxyl group; (e) Formula (II) represents an acid salt obtained by treating an acid with a base or by converting one salt into another ion exchange pathway; or A compound of formula (3) (f) wherein R and R together represent an oxo group and OR is -OH or methoxy obtained when cultured An organism of the genus Streptomyces capable of producing at least one of the said compounds. 8. A method for controlling pests in agriculture, horticultural, horticultural, forestry and other (except therapeutic treatment of humans and animals), comprising administering to the crop, other vegetation or localized pests an effective amount of one or more compounds according to claim 1 or a composition according to See point 6.