JPH07504422A - Cyclic ketal derivative - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Cyclic ketal derivative The present invention has blood cholesterol lowering, blood lipid lowering and/or antifungal activity. novel compounds, their production methods, pharmaceutical compositions containing them, medical and special for the treatment and/or prevention of atherosclerosis and related cardiovascular diseases. regarding their uses. The present invention reduces blood cholesterol, blood lipids, and and/or as intermediates for the production of compounds with antifungal activity. It also relates to compounds.

High levels of blood cholesterol and blood lipids are associated with the development of vascular wall disease. Ru. Therefore, there is great interest in effective methods for reducing plasma cholesterol levels. Leaning back. Cholesterol concentration is reduced by reducing dietary intake of sterols, e.g. by increasing its metabolism and excretion, or decreasing its rate of biosynthesis. can be reduced. Most effective in lowering physiological cholesterol levels Effective methods include - as a component, inhibition of cholesterol biosynthesis. it is Cholesterol synthesis undergoes feedback regulation, resulting in cholesterol levels This is because the decrease in biosynthesis is compensated for by an increase in biosynthesis.

One rate-limiting step in cholesterol biosynthesis is 3-hydroxy-3-methyl Formation of mevalonic acid from glutaryl coenzyme A (HMGCoA), which is associated with high cholesterol. The mebic acid group of HMG CoA reductase inhibitors is used in the treatment of rollemia. Clinical success was achieved. However, mevalonic acid has been shown to be co-fermented in animals. Common to all isoprenyl derivatives, including elemental Q, heme A and dolichols It is a precursor.

The first biosynthetic step leading exclusively to sterols, i.e. giving squalene2 The condensation of two farnesyl diphosphates is the second regulatory point. farnesirni In the synthesis of squalene from phosphoric acid, an isolable intermediate, namely presqualene The total synthetic pathway involves the membrane-bound enzyme squalene synthase (farnesinic acid). luniphosphate: farnesylniphosphate farnesyltransferase.

EC2,5,1,21). Therefore, squalene synthase Substances that act to inhibit enzymes are potential drugs for regulating cholesterol formation It is a thing. The use of such substances suggests that non-steroidal pathways are minimally affected. It is attractive because it is not.

The biosynthesis of ergosterol, a major sterol component of fungal cell membranes, is important in humans, including humans. similar to that of cholesterol in mammals, and thus squalene mediated by the intase enzyme. Therefore, fungal cells have squalene synthase Substances that act to inhibit enzymes are drugs that can be used for antifungal chemotherapy. Ru.

We act as inhibitors of the squalene synthase enzyme and/or A new intermediate for the production of compounds that act as inhibitors of quarene synthase enzymes. We have now discovered a group of novel compounds.

Therefore, in the first aspect of the invention we use the general formula %formula %: [In the above formula, R1 is a hydroxyl group, or -ococH-CHCH(CH)( CH2) 3CH3, -ocE E OCR-CHC(CH)-CHCH(CH3)CH2CH3 or -0CO-X -CH2CH(CH3)CH2CH3 (where X is -CH-CHCH(CH3 )-1-CHCH(OH)C)1(CH)-1-CH-CHC(OH)(CH )-1-CH2CH(0H)CH- or -CHCHCH(CH3)- ) represents a group selected from; R2 represents a hydroxyl group; R3 is (In the above formula, R7 is a hydrogen atom or an acetyl group), E8 -C(CH)-CHCH(CHR)CH2Ph (where R8 is hydrogen or hydrogen ), -C(CH0H)-CHCH(CH)CH2Ph.

-C(-CH)CH(OH)CH(CH20H)CH2Ph, -C(-CH)C H(NHCOCH3)CH(CH)CHPh, -C(CHNHCOCH3)- CHCH(CH3)CI (represents a group selected from 2Ph and; R and R5 each independently represent a hydrogen atom or a methyl group; R6 is attached to the rest of the molecule via a ring carbon atom and optionally one of the ring nitrogen atoms. In one C alkyl group ■-4 represents a tetrazole ring substituted with ] and salts thereof.

It is understood that all tautomers of the R6 substituent are included in the general formula (1) above. It should be.

The term C alkyl belonging to R6 refers to a straight chain or branched chain containing 1 to 4 carbon atoms. chains, more specifically methyl, ethyl, n-propyl, n-butyl, S-butyl or or t-butyl.

Compounds of formulas in which R and R5 represent hydrogen atoms or physiologically acceptable cations normally preferred.

R1 is preferably the following group: R3 is preferably the following group: (wherein R7 is a hydrogen atom or an acetyl group).

R6 preferably represents unsubstituted or methyl substituted tetrazole.

It is to be understood that the invention encompasses any combination of the above selections.

Particularly preferred compounds of formula (+) include: (Is-[1α(4R,5S*), 3α, 4β, 5N * α. 6α(2E, 4R, 6R*), 7β)) 1-(4-acetyloxy-5-methyl thyl-3-methylene-6-)dinylhexyl)-4,6,7-drihydroxy 3-(IH-tetrazol-5-yl)-2,8-dioxabicycloCB, 2. 1] Octane-4,5-dicarboxylic acid, 6-(4,6-dimethyl-2-octane noate); [15-[1α(4R,5S*), 3α, 4β, 5* α. 6α (2E, 4R, 6R"), 7β)) 1-(4* Monoacetyloxy-5-methyl-3-methylene-6-phenylhexyl'I-4 ．． 6.74lihydroxy-3-(2-methyl-2H-tetrazol-5-yl) -2゜8-dioxabicyclo(3,2,1)octane-4,5-dicarboxylic acid, 6-(4,6-dimethyl-2-octenoate): Cl5- [1α (4R, 55 pieces), 3α, 4β, 5* α, 6α, 7β)) 1-(4-acetyloxy-5-methyl-3-methylene-6 -phenylhexyl')-4,6゜7-dolihydroxy-3-(IH-tetra sol-5-yl)-2,8-dioxabincroC3,2,1)octane-4, 5-dicarboxylic acid: and [1,5-(1α (4R, 5S books), 3α, 4β, 5* α. 6α, 7β)) 1-(4-acetyloxy-5-methyl-3-methylene-6 -phenylhexyl'I-4,6゜7-dolihydroxy-3-(2-methyl- 2H-tetrazol-5-yl)-2,8-dioxabicyclo[3°2.1]o Cutane-4,5-dicarboxylic acid; and their salts and protected derivatives.

The compound of formula (1) in which R1 represents a hydroxyl group has squalene synthase inhibitory activity. It is also particularly useful as an intermediate for the production of related structures having properties.

The compounds of the invention may form salts with inorganic and organic acids and bases. Menstruation Scientifically acceptable base salts include alkali metal salts (e.g., disodium and niconica). sodium and potassium salts, including sodium salts), alkaline earth metal salts (e.g. , calcium salts) and inorganic base salts such as ammonium salts. proper organic Base salts include trialkylamines (e.g., triethylamine), dialkylamines, (e.g. dicyclohexylamine), optionally substituted benzylamine (e.g. p-bromobenzylamine), tris(hydroxymethyl)methyl Amine salts and amino acid salts (e.g. lysine and amino acid salts, including di-lysine salts) There are amine salts such as luginine salt). Examples of physiologically acceptable acid addition salts include: hydrochloride, hydrobromide, sulfate, alkyl or aryl sulfonate (e.g. , methanesulfonate or p-toluenesulfonate), phosphate, acetate, of citrate, succinate, lactate, tartrate, fumarate and maleate. There are salts derived from organic or inorganic acids such as

Other salts which are not physiologically acceptable are also useful in the preparation of compounds of formula (1); This forms another aspect of the invention.

Compounds of the invention inhibit squalene synthase enzyme and cholesterol biosynthesis Therefore, medically, especially in animals (particularly humans), plasma concentration is important. It is useful for a variety of conditions where lowering cholesterol levels would be beneficial. like this Examples of symptoms include those associated with hypercholesterolemia and hyperpoproteinemia. diseases, especially atherosclerosis and cardiovascular diseases (e.g. cardiac ischemic diseases, cerebral ischemic disease and peripheral arterial disease).

Compounds of the present invention that inhibit squalene synthase can be used in animals including humans. It may also be useful in treating fungal infections. For example, they are eg Candid a (for example, Candida albicans, Candida glabr ata, Candida parapsilosis and Candida p. seudotrop), Cryptococcus neol'.

rmans, Aspergillus Sp, (for example, Aspergillus lus flavus and Aspergillus fumigatus), Coccidioides (for example, Coccidioides 1 mm 1 t is), Paracoccidioides (for example) Paracocci dioides brasiliensis), Histoplasaa (eg. For example, 11 istoplasma capsulaturQ) or Blasto myces (e.g. Blastomyces dermatitidis) It is useful in treating systemic infections caused by They are Trichophyton, old crosporui or Epidermophyton (e.g. Tric hophyton menLograplytes, former crosporuI Ilcants or Epidermophyton fl.

It is also useful in treating local infections caused by species of C. ccosum. those are Torulopsis glabrata and Pityr.

It is also useful in treating fungal diseases caused by 5porvm ovale.

In vitro evaluation of the antifungal activity of the compounds of the present invention does not result in the growth of specific microorganisms. Determine the minimum inhibitory concentration (CMIC), which is the concentration of the test compound in the appropriate medium. This can be done by

Inhibiting squalene synthase in view of their potential in antifungal therapy The compounds of the invention are recommended for the treatment of various fungal infections in humans and animals.

Such infections include candidiasis and chronic mucosal candidiasis (e.g. oral and Fungal infections such as vaginal candidiasis), skin infections caused by fungi, skin and mucosal skin Skin candidiasis, dermatophytosis including white side and ring 1 infections, athlete's foot, paronychia, and cysts. Wind, erythema, intertrigo, fungal fluffy rash, candida vulvitis, candida balanitis and I have otitis externa.

They may also be useful as prophylactic agents to prevent systemic and local fungal infections. prevention Use as a selective intestinal decontamination method, e.g. in the prevention of infection in immunocompromised patients. Suitable as part of. Prevention of fungal overgrowth during antibiotic treatment may also be helpful in some medical conditions. or may be desirable in iatrogenic conditions.

Ability of Compounds of the Invention to Inhibit Squalene Synthase Enzyme in Mammals and Fungi Power is R, M, Talt, Analyt, Bjoches, 203, 310-3 18 (1992) as a substrate under assay conditions similar to those described in 2C] fufurnesyl pyrophosphate in vitro.

For therapeutic purposes, compounds of the invention that inhibit the enzyme squalene synthase may be administered directly. preferably as an active ingredient in a pharmaceutical formulation. . For this reason, the present invention provides compounds of the present invention that inhibit the squalene synthase enzyme. more than one pharmaceutically acceptable carrier and optionally other treatments and/or prophylaxis. Further provided are pharmaceutical formulations comprising the ingredients. The carrier is not included in the other components of the formulation. must be 'acceptable' in the sense of being compatible with the substance and not harmful to its receptors. do not have.

The compositions of the invention include oral, buccal, parenteral, implantable, rectal, topical, ophthalmic or Forms prescribed for genitourinary administration or suitable for administration by inhalation or insufflation There are compositions that have

Tablets and capsules for oral administration may contain binders such as syrup, acacia, gelatin, etc. of latin, sorbitol, tragacanth, starch or polyvinylpyrrolidone. Slime: fillers such as lactose, sugar, microcrystalline cellulose, maize starch , calcium phosphate or sorbitol: lubricants such as magnesi stearate. um, stearic acid, talc, polyethylene glycol or silica, disintegrant, e.g. potato starch or sodium starch glycolate; wetting agents e.g. It may also contain conventional excipients such as sodium lauryl sulfate. Tablets are in our industry It may be coated according to a well-known method. Oral liquid preparations may be e.g. aqueous or oil-based. It may be in the form of a suspension, solution, emulsion, syrup or elixir; may be served as a dry product that is prepared with water or other suitable vehicle before use. stomach. Such liquid preparations may contain suspending agents such as sorbitol syrup, methyl cell loin, glucose/sugar syrup, gelatin, hydroxyethylcellulose, carbonate ruboxymethylcellulose, aluminum stearate gel or hydrogenated edible fats; emulsifiers such as lecithin, sorbitan monooleate or gum arabic; Non-aqueous vehicles (which may also include edible oils), such as almond oil, fractionated coconut oil , oily esters, propylene glycol or ethyl alcohol; preservatives, e.g. Also contains conventional additives such as methyl or propyl p-hydroxybenzoate You may do so. The composition may include conventional herbal medicines such as cocoa butter or other glycerides. It may also be formulated as a suppository containing a base.

For oral or vaginal administration, the compositions may take the form of conventionally formulated tablets or lozenges. It's okay.

Compositions according to the invention may be formulated for parenteral administration by injection or continuous infusion. stomach. Injectable formulations are packaged and stored in unit dose form in ampoules or multi-dose containers. Agents may also be added. The compositions can be prepared as suspensions, solutions or solutions in oily or aqueous vehicles. It may be in the form of a emulsion and may contain suspending, stabilizing and/or dispersing agents. It may also contain prescription agents. On the other hand, the active ingredient must be combined with a suitable vehicle, e.g. It may also be in powder form prepared in sterile, pyrogen-free water.

For administration by inhalation, the compositions according to the invention may be administered using a suitable propellant, such as dichlorodifluoride. lomethane, trichlorofluoromethane, dichlorotetrafluoroethane, dioxide from a pressurized bag or with a nebulizer under the use of carbon or other suitable gas It is preferably delivered in the form of an aerosol spray dispensing. Place of pressurized aerosol In such cases, the dosage unit may be determined by providing a valve to deliver a metered amount.

On the other hand, for inhalation administration, the composition according to the invention may be a dry powder composition, e.g. and in the form of a powder mix with a suitable powder base such as lactose or starch. You can. Powder compositions can be packaged in capsules or cartridges, such as gelatin. , or as a blister pack in which the powder is administered with the help of an inhaler or ventilator Can be provided in unit dosage form.

The composition may be, for example, a suppository containing a conventional herbal base, or, for example, a conventional PET. It may also take the form of a pessary containing a sari base.

Compositions include ointments, creams, gels, lotions, shampoos, powders (spray powders) pessaries, tampons, sprays, dips, aerosols, layering agents (e.g. , eye, ear or nose preparations) or for topical administration in the form of a pour-on. May be prescribed. Ointments and creams may contain suitable thickening and/or gelling agents. In addition, they may be formulated with an aqueous or oily base, for example. Ointments for ocular administration are sterile Manufactured aseptically using ingredients. Pour-ons may include prescription drugs, e.g. For example, they are formulated for veterinary use in oils containing organic solvents along with stabilizing and solubilizing agents.

Pessaries and tampons for membrane seeds are formulated using conventional techniques and are If so, it contains an effervescent vehicle. Such compositions may optionally contain corticosteroids. It may also contain other active ingredients such as antibiotics, antibiotics, or anthelmintics.

Liquid formulations for intranasal delivery take the form of solutions or suspensions and may include tonicity adjusting agents, e.g. Sodium, dextrose or mannitol; preservatives such as benzal chloride Conium, thiomersal, phenylethyl alcohol; other prescription agents, e.g. It may also contain conventional excipients such as clouding, buffering, stabilizing and/or dispersing agents.

Transdermal administration facilitates absorption of the active compound through the skin and typically includes a backing film. Adhesive stick-on (stjck-o) containing adhesive film, membrane and release liner n) This can be done by designing a suitable system consisting of encapsulated base formulations within a batch.

Compositions according to the invention may be formulated as devo preparations.

Such long-acting formulations may be implanted (e.g. subcutaneously or intramuscularly) or intramuscularly. Administered by injection. Thus, for example, the compounds of the present invention may be combined with suitable polymers. or hydrophobic substances (e.g. as emulsions in acceptable oils) or ions. It is formulated with exchange resins or as poorly soluble derivatives, such as poorly soluble salts.

When the composition consists of dosage units, each The unit is preferably 0.001 to 1000 mg, advantageously 0.01 to 400 mg Contains active ingredients. Daily doses as used in adult therapy may be determined by e.g. route of administration. and depending on the patient's condition and the disease being treated, preferably from 0.001 to 0.001 of the active ingredient. 5000 mg, most preferably within the range of 0.01 to 2000 l1g , administered 1 to 4 times a day.

The compound can be administered by intravenous infusion using, for example, up to 50 mg/kg/day of active ingredient. more administered. Treatment duration is determined by response rate rather than an arbitrary number of days. It will be done.

Compounds of the invention that inhibit squalene synthase may be used in combination with other therapeutic agents. Therefore, the present invention utilizes compounds of the present invention that inhibit squalene synthase in other treatments. Activators, such as 3-hydroxy-3-methylglutaryl coenzyme A (HMG Co A) Inhibitors of reductase or drugs that reduce serum cholesterol and/or or other drugs that inhibit cholesterol biosynthesis, such as bile acid sequestrants, or Antihyperpoproteinemic or antihyperlipidemic agents, such as probucol, gemfi Brozil, clofibrate, dextrothyroxine or its sodium salt, Cholesterol or its hydrochloride, cholestyramine, nicotinic acid, neomycin , p-aminosalicylic acid, aspirin, DEAE-Sephadex, poly(dia (lylmethylamine) derivatives, ionene or poly(diallyldimethylammonium) In another aspect, the combination is provided with chloride.

The above combination is preferably put into use in the form of a pharmaceutical formulation, for which Pharmaceutical formulations containing the combination together with a pharmaceutically acceptable carrier are another aspect of the invention. form a surface. The individual components of such combinations may be administered separately or in combination. Administered sequentially or simultaneously.

When a compound of the present invention is used in combination with a second therapeutic agent for a similar condition, each compound The dosage of the compound may be different than when the compound is used alone. appropriate use Amounts will be readily apparent to those skilled in the art.

According to another aspect of the invention we provide a compound of formula (1) or its physiologically acceptable salts that can be tolerated or for therapeutic use, especially plasma cholesterol in animals (especially humans) Treatment of conditions where lowering of drug levels is beneficial or true in animals (particularly humans) The compound of formula (1) as described above or its physiologically acceptable form for the treatment of fungal infections. The present invention provides a pharmaceutical composition containing a salt of the present invention.

In a specific aspect of the invention, we provide a compound of formula (1) or its physiologically acceptable or hypercholesterolemia and/or hyperpoproteinemia diseases related to heart disease, especially atherosclerosis and cardiovascular disease (e.g. heart failure). Formula (1) as above for the treatment of blood diseases, cerebral ischemic diseases and peripheral arterial diseases) or a physiologically acceptable salt thereof.

According to another aspect of the invention, we provide a method for treating hypercholesterolemia and/or high blood pressure. Diseases associated with proteinemia, especially atherosclerosis and cardiovascular diseases ( For example, for the production of drugs for the treatment of cardiac ischemic disease, cerebral ischemic disease, and peripheral arterial disease. The use of a compound of formula (+) or a physiologically acceptable salt thereof is provided.

According to another aspect of the invention, we provide a method for treating fungal infections in human or non-human animal patients. Compounds of formula (1) or physiologically acceptable compounds thereof for the manufacture of drugs for the treatment of infections Provides for the use of salt.

According to another aspect of the invention, we provide a method for treating hypercholesterolemia and/or high blood pressure. Diseases associated with proteinemia, especially atherosclerosis and cardiovascular diseases ( For example, to treat cardiac ischemic disease, cerebral ischemic disease and peripheral arterial disease) or to treat fungal Provided is a method of treating a human or non-human animal individual for treating a disease; The method includes administering an effective amount of the compound of formula (1) or a physiologically acceptable salt thereof to the above individual. It consists of administering.

When we refer to treatment here, we refer to prevention as well as treatment of established symptoms or infections. It will be clear to those skilled in the art that this also applies.

The compounds of the present invention are produced by the following process.

General procedure for producing compounds of formula (1) in which R6 is an unsubstituted tetrazole ring Seth (A) is a compound of the following formula (11): (In the above formula, R-R3 has the same meaning as above, and R4a and R5a are protecting groups. ) at high temperature (e.g., about 120"C) in a solvent such as dimethylformamide. preferably the presence of an organic base or a salt thereof (e.g. triethylamine hydrochloride) of sodium azide or azidotrimethylsilane, optionally in the presence of water. treatment with an azide source such as, followed by removal of any protecting groups present if necessary. It will be.

The compound of formula (II) is a compound of the following formula (II+): (In the above formula, R-R, R and and R5a have the same meanings as above 1 3 4a ) is produced by dehydrating it.

Dehydration is carried out under conventional conditions, for example by treating the compound of formula (11) with an organic base (e.g. halogenated hydrocarbons (e.g. dichloromethane) in the presence of treatment with an acid anhydride such as trifluoroacetic anhydride in a solvent such as (or, if pyridine is the base, this can represent the solvent) good. The reaction is preferably carried out at about room temperature.

The compound of formula (II+) has the following formula (compound of m: 1 3 4a (In the above formula, R-R, R and R5a have the same meanings as above), and 3- It is produced by converting a carboxyl group to a carboxamide group.

The reaction preferably involves activation of the 3-carboxyl group followed by ammonia under conventional conditions. Rell done by processing with Nia.

Thus, for example, amination preferably activates the activated derivative of the compound of formula (1v). This is carried out, for example, by treatment with ammonia gas at a temperature of 0 to 20'C.

Activation of the 3-carboxyl group can be carried out, for example, with a suitable halogen in dimethylformamide. hydrocarbons (e.g. dichloromethane), ethers (e.g. tetrahydrofuryl) (e.g. acetonitrile) or a nitrile (e.g. acetonitrile). and preferably by reaction with a reagent such as oxalyl chloride at a temperature of about 0°C. be exposed.

The compound of formula (II) is produced by producing the compound of formula (!1) from the compound of formula (II+). Compounds of formula (V) below using the conditions described above for: (In the above formula, R, R3, R4a and R5a have the same meanings as above, and R2a is R2 or is a protected hydroxyl group) and then, if present, It is also produced by removing the hydroxyl protecting group.

The compound of formula (v) is a compound of the following formula (vl): (in the above formula, R, R, R2aS R4a (J:R” has the same meaning as above) is hydroxylamine or a salt thereof (e.g. hydrochloride). The reaction preferably It is carried out in a suitable solvent such as lysine at about room temperature. Hydroxylamine hydrochloride If a salt such as is used, the reaction is carried out in the presence of a base. suitable bases include There is pyridine, which can also be used as a reaction solvent.

The compound of formula (vl) is obtained by activating the 3-carboxyl group from the compound of formula (1v). , followed by an amide (e.g. dimethylformamide) or an ether (e.g. trahydrofuran) at a suitable temperature, e.g. borohydride (e.g., sodium borohydride) at about room temperature). It is produced by reduction with a suitable reducing agent such as

Activation of the 3-carboxyl group can be performed, for example, by using an ether (e.g., tetrahydrofuran). Carbodiimides (e.g. For example, 1-cyclohexyl-3-(2-morpholinoethyl)carbodiimide meth -p-toluenesulfonate or N,N--dicyclohexylcarbodiimide ) or by reaction with a reagent such as N-hydroxysuccinimide in the presence of or a halogenated hydrocarbon (e.g. dichloromethane). 2-C in the presence of a non-nucleophilic organic base such as ethylamine at a temperature ranging from 0 to 20 °C. By reaction with lolo-3-ethylbenzoxacilium tetrafluoroborate This is done by conversion to an active ester. On the other hand, activation is performed using dimethyl fluoride as described above. Compounds of formula (vl) can also be prepared by reaction with oxalyl chloride in lumamide. Compound of the following formula ml: 1 3 4a (In the above formula, R, R, R and R5a have the same meanings as above, and R9 is hydroxyl oxidizing the hydroxyl protecting group) and then removing the hydroxyl protecting group if desired. It may also be manufactured by The oxidation is N~methylphorphorin in the presence of N-oxide, Perruthenate (e.g. A catalytic amount of an oxidizing agent such as tetra(n-propylammonium perruthenate) It is done using Suitable solvents for oxidation include nitriles such as acetonitrile. , the reaction is preferably carried out at about room temperature. On the other hand, the oxidation is preferably carried out using anhydrous truffles. halogenated hydrocarbons (e.g. dichloromethane) in the presence of fluoroacetic acid. A sulfoxide such as dimethyl sulfoxide in a solvent at low temperature (e.g., about -70°C) This is done using Sid.

The compound of formula (Vl+) can be used for the production of the compound of formula (vl) from the compound of formula (IV). Under the reducing conditions described above for the corresponding acid of formula (■111): 1 B 4a 5a (In the above formula, R, R, R, R and R9 have the same meanings as above.)

Compounds of formula (1v) and ml+) are preferably prepared using standard carboxylic acids and hydroxides. Compound of the following formula (1x) by the roxyl retention/deprotection method: (In the above formula, R-R3 has the same meaning as above.)

Compounds of formula (IV) and (Vll+) can be prepared in a suitable salt such as sodium hydroxide. of the following formula (X) by saponification of the 3-position carboxylic acid methyl ester group using Compound: 1 3 4a (In the above formula, R-R, R and R5a have the same meanings as above).

Compounds of formula (X) are prepared from compounds of formula (IX) using conventional esterification conditions. be done.

Another process (B) for the production of the compound of formula (1) is the production of the compound of formula (1) Converting a compound or a protected derivative thereof into a different compound of formula (1) or a protected derivative thereof and then, if necessary, removing any protective groups present.

Therefore, in embodiments of process (B), R6 is an alkylated tetrazole ring. A compound of formula (1) representing a compound of formula (1) in which R6 represents an unsubstituted tetrazole ring From the corresponding carboxylic acid protected derivatives, alkylation is then present if necessary. Produced by removal of protecting groups. The alkylation reaction is carried out under conventional conditions, e.g. inorganic Dimethylforma in the presence of a base such as a carbonate (e.g. sodium bicarbonate) By treating the protected intermediate with C alkyl iodide in a solvent such as Mido at about room temperature, Manufactured by

In another embodiment of process (B), the formula (+ ) can be prepared using the general deacylation conditions described below with R1 of formula (1 ) Deacylation of corresponding compounds of formula (1) representing an acyloxy group as described above in Manufactured by.

Carboxylic acid and hydroxyl protecting groups suitable for use herein include, for example, ° Protective Groups in Organic CheIIli stry’, Edl, F, W, McOmie (Plenum Press, 19 73) or °Protective Groups in Organic 5ynthesis’by Theodora w, Greene (John Wiley and 5ons, 1991) There are protective groups. Examples of suitable carboxylic acid protecting groups include methyl or t-butyl Alkyl groups such as thyl, 2-methoxyethoxymethyl, or diphenylmethyl There are aralkyl groups such as thyl or p-nitrobenzyl. appropriate hydroxy Examples of silyl protecting groups include 2-methoxyethoxymethyl and silyl groups (e.g. t-butyldimethylsilyl).

Protecting groups are removed using conventional techniques. For this reason, for example, t-butyl The alkyl group (e.g. ether, hydrogen chloride in a solvent such as dioxane) ) under anhydrous acid conditions. On the other hand, removal of the methyl protecting group Lithium iodide at elevated temperature in rusulfoxide or 2,4,6-drimethylpyridine This is done using The p-nitrobenzyl group is preferably an ether (e.g. zinc metal or aqueous tetrahydrofuran). and hydrochloric acid. diphenylmethyl group or 2-methoxyethoxy Methyl groups are preferably removed using aqueous formic acid or trifluoroacetic acid.

Silyl groups such as t-butyldimethylsilyl are preferably treated with fluoride ions. removed.

Esterification of the carboxylic acid group of a suitable intermediate compound to the corresponding methyl ester group is a preferred method. Preferably an amide (such as (f1 dimethylacetamide or preferably dimethylfotamide) Lumamide methyl halide (e.g., iodine) in the presence of a base such as sodium bicarbonate). methyl chloride) or by treatment with a methylating agent such as dimethyl sulfate.

Reaction C is preferably carried out at a temperature in the range of 0 to 100°C, preferably 20 to 30°C. Ru. On the other hand, esterification is the esterification of diazomethane in a suitable solvent such as methanol. The treatment may also be carried out by treatment with a hydrophobic solution. Esterification it, ri + * ore at room temperature Also by treatment with methanol in the presence of a suitable acid such as hydrochloric acid I can do it.

The conversion of one methyl ester to a different methyl ester requires appropriate esterification/de-esterification. It will be carried out from Teruka Ste Nobu 1. Deesterification was carried out under standard conditions with flJ( By f-base calohydrolysis, or aqueous dimethyl sulfoxide is It is carried out using Euonymus ninotium in remethylpyridine at elevated temperature.

Compounds of formula (1x) may be prepared by the fermentation process described below or by a suitable aliquot. Sylation and deacylation method 1 (Acylation or deacylation at position 6 as appropriate) It can also be manufactured from the product of the chemical process (1). Law C is described below. Deacylation is preferably carried out using an alcohol (e.g. Base using a base like aqueous sodium hydroxide in a solvent like methanol) It is carried out by catalytic hydrolysis. On the other hand, deacylation of α,β-unsaturated esters is Optionally in a solvent such as dimethylformamide with a suitable base (e.g. triethyl hydroxylamine (e.g. trialkylamine) in the presence of a trialkylamine such as , N-methylhydroxylamine hydrochloride). In the fermentation process Cultivating a microorganism capable of producing one or more suitable compounds of formula (IX) . The desired compound is then isolated from the culture and acylated or Deacylated and/or esterified to the corresponding methyl ester.

Suitable microorganisms can be used to test samples obtained from microbial fermentation using small scale tests. is easily confirmed by analysis using standard methods.

In particular, the commonly used microorganism is CAB International Mycol.

gical Institute, Ferry Road. Kev. Surr. This is a strain of the microorganism deposited with E.EY, England. The stock was released in May 1989. It was accepted by the institution on the 25th, and then the accession number! Old 332962 and 1989 The deposit date was June 27th (the date of confirmation of survival). The deposited stock has accession number 1 13329B2 in this specification. I this regarding old 332962 The characteristics confirmed up to this point are shown in Example 15 below.

It is clear that the desired intermediate may also be produced from the IM+ 332962 mutant strain. Will.

Mutant strains of 1M1 332962 occur naturally or and Radioisotopes for Industrial Mi croorganisss'. Proceedings of the Sym posium. Vienna. 1973, p241.

+nternattona+ Atomic Energy Authorit ylkoo (AteH.1.

Techniques for the Developem by Adler A variety of microorganisms, including those described in methods such as ionizing radiation, chemical methods, e.g. N-Methyl-N-m = treatment with tro-N nitrosoguanidine (NTG), processing heat, genetic techniques, e.g. recombination and transformation and selective techniques for natural variants; There is.

Fermentation is carried out by conventional means, i.e. in the presence of assimilable sources of carbon, nitrogen and inorganic salts. This is done by culturing.

Assimilated sources of carbon, nitrogen and minerals are provided by simple and complex nutrients.

Carbon sources typically include glucose, maltose, starch, glycerol, molasses, and dextrose. Sto1non, lactose, sucrose, fructose, galactose, myoino Sitol, D-mannitol, soybean oil, carboxylic acid, amino acids, glycerides, amino acids Luconol, 7/lz can and vegetable oil. The carbon source is usually the fermentation medium. and sucrose are the preferred carbon sources.

Nitrogen sources typically include soybean meal, corn steeping liquid, distiller's soluble ingredients, yeast extract, Cottonseed meal, peptone, ground nut meal, malt extract, molasses, casein, aminos acid mixtures, ammonia (gas or solution), ammonium salts or nitrates. Ru. Urea and other amides may also be used. The nitrogen source is usually 0.1 to 1 in the fermentation medium. It is 0% by weight.

Nutrient inorganic salts added to the culture medium include sodium, potassium, ammonium, Iron, magnesium, zinc, nickel, cobalt, manganese, vanadium, chromium , calcium, copper, molybdenum, boron, phosphoric acid, sulfuric acid, chloride and carbonate ions There are commonly used salts that can produce ions.

Cultivation of organisms is usually carried out at 20-40°C, preferably at 20-35°C, particularly at about 25-28°C. temperature and may be carried out by aeration and agitation, e.g. shaking or stirring. desirable. The medium is first inoculated with a small amount of mycelia and/or spores. obtained The nutrient inoculum is either a fermentation medium or subjected to further growth before being transferred to the main fermentation medium. transferred to one or more seed stages. Fermentation is usually 3.5-9.5, preferably 4.5-7 ．． It is carried out in a pH range of 5. To keep the pH within the desired range, the fermentation medium must be It is necessary to add a base or acid. Suitable bases to add include aqueous There are alkali metal hydroxides such as sodium hydroxide or potassium hydroxide. suitable Correct acids include mineral acids such as hydrochloric acid, sulfuric acid, or phosphoric acid.

Fermentation is carried out over a period of 4 to 30 days, preferably about 7 to 18 days. Defoaming Agents may also be present to control excessive foaming and should be added at the required intervals. It will be done.

Carbon and/or nitrogen sources may also be provided into the fermentation medium if necessary.

The products of the fermentation process are present both in the fermentation liquor and in the mycelial fraction, preferably filtered. Or separated by centrifugation.

Optionally, the solution is then cooled until the pH is below pH 6 (e.g., about pH 3). treated with an acid such as sulfuric acid in the presence of an organic solvent.

Products of the fermentation process are separated from the fermentation broth by conventional isolation and separation techniques. You may. It will be clear that the choice of isolation technique is wide-ranging.

The products of the fermentation process are subjected to various fractionation techniques, such as adsorption-elution, sedimentation, fractional crystallization, Isolated and purified by solvent extraction and liquid-liquid partitioning, which can be You can combine them either way.

Adsorption to a solid support followed by elution allows the compounds of the invention to be isolated and It was found to be suitable for purification.

The products of the fermentation process are ketones (e.g. acetone, methyl ethyl ketone or methyl isobutyl ketone), halogenated hydrocarbons, alcohols, diols (e.g. propane-1,2-diol or butane-1,3-diol) or Cells are washed with a suitable organic solvent such as methyl acetate or ethyl acetate (e.g. methyl acetate or ethyl acetate). and extracted from the aqueous phase. Typically, two or more extractions will help achieve the best recovery. desirable. The water-immiscible solvent extract itself is extracted with a basic aqueous solution, and these basic After acidification of the solution, the desired compounds are re-extracted into the water-immiscible organic phase.

Removal of the solvent (e.g., by evaporation) from the organic extract containing the desired compounds Obtain the substance.

Chromatography (including high performance liquid chromatography) Suitable supports; non-functional cathode adsorption resins, e.g. Amberllte XAD- 2. XAD-4, XAD-16 or X, AD-1180 resin (Rohm & Baas Ltd) or KaStell S112 (Montedison ); cross-linked styrene-divinylbenzene polymer resins such as substituted styrene-divinylbenzene Rubenzene polymers, such as halo such as Diaion 5P207 (Mitsubishi) Geninated (e.g., brominated) styrene divinylbenzene polymer; anion exchange agent (For example, IRA-35 or IRA-48); 5ephadex LH20 Organic solvent compatible cross-linked dextrin such as (1’harmacia LIK Ltd) Tran; carried out on a reversed phase support such as hydrocarbon-bonded silica, e.g. C18-bonded silica. be exposed. Another chromatographic means for purification/separation of products of fermentation processes is countercurrent chromatography using a coil extractor, such as a multilayer coil extractor. be.

The products of the fermentation process can be isolated or isolated by the use of liquid anion exchangers such as LA2. You can also purify it.

When IRA-68 or especially IRA-35 is used as a solid adsorbent, the cell The extract is drained directly without removal of the solvent.

The extract may be added directly at about pH 3 or about pH 8 after filtration of solid impurities.

Suitable solutions for chromatographic purification TA/separation of suitable compounds of formula (IX) The medium/eluent will of course depend on the column type and the nature of the support. Countercurrent chromatography When using chemistries, we use ethyl acetate, hexane, methanol and aqueous Solvent systems containing acids (e.g. aqueous sulfuric acid) have been found to be particularly suitable.IRA When using an anion exchanger such as -35, the tree strength is preferably and then eluted with sulfuric acid in aqueous acetone.

The presence of the products of the fermentation process during the extraction/isolation operation is determined by h, p, 1. c, also by conventional techniques such as UV spectroscopy or by using the properties of the compound. What to do: Be monitored by others.

Products of fermentation processes, e.g. dissolved in organic solvents after chromatographic purification If the desired compound is obtained in the form 1. (i) by evaporation. If desired, the compound can be removed by chromatography as described above. further refined.

A compound of formula (IX) in which R1 represents an acyloxy group as described above in formula (+) The acylation given can be achieved by formula (IX ) with a suitable acylating agent under conventional esterification conditions, This is carried out by subsequently removing the protective groups present. For this reason, for example, the expression ( When R1 in IX) represents, the R group is a formula (IX) in which R1 is a hydroxy group. A compound of the following formula (xl): or by treatment with an activated derivative thereof. For this reason, the formula (XI ) is preferably acylated with an acid such as dicyclohexylcarbodiimide. a suitable salt such as 4-dimethylaminopyridine in the presence of a suitable carbodiimide. In the presence of the group, a solvent such as a /10 generated hydrocarbon (e.g. dichloromethane) It takes place inside. On the other hand, the acid of formula (xl) can be prepared using, for example, thionyl chloride to obtain the corresponding acid chloride. 1, and then the acylation reaction is performed with /10 hydrogenated carbon f arsenide (e.g. 2.4°6-trimethylpyridine or N,N- in a solvent such as chloromethane) Alkali metal carbonate or carbonate in the presence of a base such as dimethylaniline or It is carried out using alkaline earth metals (eg calcium carbonate).

Acylation or deacylation and esterification processes may be carried out sequentially or simultaneously, as appropriate. It is understood that the steps may be combined as appropriate.

The compound of formula (Xl) is preferably a hydrolyzed compound of formula (IX) in which R1 represents By solution, aqueous hydroxyl in a solvent such as alcohol (e.g. methanol) It is produced by base-catalyzed hydrolysis using a base such as sodium chloride.

The base salt of the compound of formula (1) is preferably a base salt of the compound of formula (1). formed by treatment with a group. For this reason, for example, salt is preferably with sodium or potassium hydroxide, bicarbonate, carbonate or acetate (e.g. , potassium hydroxide, potassium bicarbonate, sodium bicarbonate or potassium acetate ), ammonium acetate, calcium acetate and L-lysine; is prepared by treating a compound of formula (R) with a base. a suitable solvent and/or alcohol (e.g. methanol) or nitrile. solution of the compound of formula (+) in a co-solvent such as acetonitrile or The suspension is treated with suitable salts or salts at a temperature of e.g. 0 to 80°C, preferably at about room temperature. or by adding a base (if necessary as an aqueous solution).

Acid addition salts can be prepared in a suitable solvent such as water and/or alcohol (e.g. methane). in the presence of a co-solvent such as a nitrile (e.g. acetonitrile) It is produced by treating a compound of formula (1) with a suitable acid.

Physiologically acceptable salts can be prepared using conventional methods using other physiologically acceptable salts of the compound of formula (1). It may also be made from other salts, including those salts.

Compounds of formula (11) and (V) are novel intermediates and form another aspect of the invention. Ru.

The following examples are given to illustrate the invention and are not intended to limit it in any way. isn't it.

Intermediate 1 (Is-[1α(4R,5S*), 3α, 4β, 5α.

* Monotricarboxylic acid, 6-(4,6-dimethyl-2-octenoate) (a) IMl 3329B2 was grown on agar plates with the following composition: Malt extract (Oxold R39) 30g Fungal peptone (Oxoid R40) ) 5g yeast extract (Oxoid R21) 0.5g agar (Oxoid No. , 3) 20g distilled water, total amount up to 1 liter The pH of the medium before autoclaving was in the range of 5.3 to 5.5. Contact The seeded plates were incubated at 28°C for 14 days. Covered with fungal mycelium Several 6-diameter plugs of agar were cut out from the growth end of the culture, and the two plugs were was placed in each of several cryotubes containing 1.6 l of sterile distilled water. the The tube was capped and stored at room temperature until needed.

contained in a 250a+l Erlenmeyer flask using two agar plugs. 501 seed medium (A) was inoculated into 8 cells each: Seed medium (A): Peptone (Oxoid R34) 10g malt extract (Oxoid d R39) 21g glycerol 40g Junlon 110 (Honeywill & 5tein) Ltd, Wa llington, 5urrey) 1g distilled water, total amount up to 1 liter The pH of the medium was adjusted to 6.3-6.5 with aqueous sodium hydroxide before autoclaving. It was adjusted.

The flask of inoculated seed medium was heated at 250 rpa+ while moving in a 50 mm diameter orbit. The cells were incubated for 5 days at 25° C. on a shaker plate rotating at 50°C.

The contents of the flasks were pooled and homogenized. homogenized seed culture was used at 3% (V/V) to prepare fermentation medium (B ) 120 pieces were inoculated with 50 al each: Fermentation medium (B): Glycerol 50g Soybean oil 30g Cottonseed flour (Sigma) 10g Distilled water up to 1 liter The pH of the medium before autoclaving was in the range of 6.1 to 6.3. centre The flasks were incubated as above for 8 days with shaking.

Fermentation broth (approximately 6 L) from flasks incubated for 8 days was filtered to remove bacteria. The strands were removed and the filtrate was adjusted to pH 2.8 with sulfuric acid (20% v/v) and diluted with double volume. Extracted three times with ethyl acetate. Combine the ethyl acetate extracts and dilute with aqueous sodium bicarbonate. The mixture was back-extracted twice with 400 ml of a 1% v/v solution. Combine the aqueous back extract; The pH was adjusted to 2°8 as before and re-extracted twice into 50 ml of ethyl acetate. .

These extracts were combined and evaporated to dryness to give a brown oil. This oil is It o Mul Law 1ayer Co11 ExLracLor (P, C, Inc, , Potomac, Maryland, USA) Further treatment was performed with tography. The coil used has a total capacitance of approximately 3801 Standard prevariety consisting of 70 meters of 2.6mm inner diameter PTFE tubing It was a coil. The solvent system used was ethyl acetate, hexane, methanol and It was a mixture of N/100 sulfuric acid (6:5:5:6 by volume). The lower phase remains unchanged. Ta. G11son Model 303 pump and Model 80 on the coil 4CManometricModule (Gilson, Vlliers L e Be1. France) and filled with the lower phase. Then oil (upper phase 41 +497 mg in lower phase 41) was injected at the "tail" end of the column. Then centrifuge was operated at 800 revolutions/min, and the mobile (upper) phase was transferred from the “tail” end of the column to Pumping was carried out at ml/min. The 201 fractions were collected, and squalene synthase inhibition was performed. Monitored by measuring harm.

Consecutive fractions showing activity against squalene synthase were combined. Steaming the early fractions The mixture was evaporated to dryness to give the title compound (90 mg) as a pale yellow oil.

(b) Mycelium isolated from 6L broth for 8 days according to the procedure in part (a) above. Extract the incubated flask with methanol (2x 3 L) and filter. did. The filtrate was concentrated to about 500 ml by evaporation, adjusted to pH 3.0 with formic acid, Extracted three times with ethyl acetate 5001. Combine the ethyl acetate extracts and dilute with sodium bicarbonate. Back-extraction was carried out twice with Lim solution (1% v/v) 2001. Combine the aqueous back extract; The pH was adjusted to 3.0 and re-extracted twice into 500 sl of ethyl acetate. all organic The fractions were combined and reduced to dryness using a rotary evaporator to remove the brown oil. Obtained. The oil (578II 1g) was collected using three G11son solvent delivery pumps (Model 578II). Dell 303), 811 Dynamic Mixer and 802CManomet G11son autopreparative system consisting of ric Module The sample was further processed by high performance liquid chromatography (HPLC) using a solvent. chroma The toography was performed using Dynamax Microsorb C10 (5 μm) semi-particles. It was carried out on a reparative column (250×10III11). Mobile phase for 65 minutes Pumped at 2.8-5, 61111/ff1in over a run time of acetonitrile and p) 0.1% v/v formic acid and ammonium acetate up to i3.15. um (1:3-44:1→1:3). This method 1 Repeated 6 times. The 13X4.95m1n fraction was collected and evaluated for inhibition of squalene synthase. Monitored by measuring harm.

Fraction No. 5 from each run was combined, acidified to pH 3.0 with formic acid, and ethyl acetate Extracted twice with 100ml. The organic phase was removed and evaporated to dryness as a pale yellow oil. The title compound (172 mg) was obtained.

(c) (i) 0.5 ml from the 5-day fermentation carried out as in part (a) above. Seed medium (A ) 50 ml each was inoculated into 8 cells. While moving the flask in a 50) radial orbit, 5. Incubate at 25°C for 4 days on a shaker plate rotating at 5.0 rpm. Ta. The contents of the flasks were pooled and homogenized.

Using a homogenized seed culture at 3% (V/V), Fermentation medium (B) 120 pieces each of 50I111 were inoculated in Mayer flasks. Hula The scions were incubated with shaking as above for 10 days.

(c) (i) Homogenized seed culture prepared as in part (c) (H) above. Fermentation of 5 liters and 3 liters each using nutrients at 3% (V/V) Two fermentation vessels containing medium (B) were inoculated. The inoculation medium was maintained at 25°C; Stirred with two six-sided turbines rotating at 500 rpm (70mm diameter) . The culture was aerated by sparging with 3 LprH of sterile air. Silico Overexposure of the culture was prevented by the addition of antifoam agent (Dov Corning 1520). I prepared to control the bubbles. After 11 days of growth, the contents of two culture vessels were combined and further treated by countercurrent chromatography according to the procedure of part (a) above, The title compound (137B) was obtained with 1 deuterium-500 MHz proton in methanol. N, +n, r.

Then, about 60.84-0.90 (+n, 9tl), 1.03 (d, 7) , 3H), 1, 09-1.19 (m, 2H), 2.10 (s, 3) 1 ), 2.24 (m, LH), 2J4 (m, 1.H), 2.68 ( dd, 13.6.1H), 4.04(d, 2.IH), 4.97(s , 1) 1), 5.02 (s, IH), 5.08 (d, 5. LH ), 5.27 (s, LH), 5.80 (d, 18.18), 6.3 1 (d, 2.111), tli, 85 (dd, 1U゜ 8, IH), 7.14 (t, 7. LH), 7.19 (d, 7, 2H) , shows a signal at 7.26 (t, 7.2H): in deuterium-methanol In the composite pulse decoupling 125.75MHz carbon-13 N, m, r, Approximately δ172, 5 (0), 172.1 (0), 170.1 (0), 168.5 (0), 166, 5 (0), 157.8 (1), 147. 7 (0), 1416 (0), 130.2 (1), 129.3 (1 ), 126.9 (L), 119,8 (1), 111.5 (2), 106.8 (0), 91.1 (0), 82.5 (1), 81.0 (1), 80.1 (1), 76.6 (1), 75.6 (0), 4 4.4 (2), 40.9 (2), 37.7 (L), 35.6 (1) , 34.9 (2), 3'3.1 (L), 30.8 (2), 26. 5 (2), 20.9 (3), 20.5 (3), 19.2 (3), 14. Peaks are shown at 1 (3) and 11.4 ('3).

(d) (+> Frozen stock of inoculum for 5 days carried out as in part (a) above) Prepared from fermentation. Centrifuge the culture sample for 10 minutes and remove the mycelium from 15% glycerinase. Roll and resuspended in 0.01% Tween 80 to the original amount. mycelium Spin down and resuspend again, then add 1.8ml into a plastic tube. Amounts were distributed and stored at -20°C. Frozen inoculum 0. Using 8 pieces each of 5+nl , 50 ml each of seed medium (A) contained in 2501 Erlenmeyer flasks. Eight were inoculated. Rotate the flask at 25 Orpm while moving in a 50-diameter orbit. The cells were incubated for 4 days at 25°C on a shaker plate. Inside the seed flask The contents were pooled and used at 3% (V/V) in a 250 ml Lullenmeyer flask. 50 ml of fermentation medium (B) was inoculated into 120 cells each. the flask for 9 days. (d) m) above bar) (d) (H) The contents of the four end-stage flasks were pooled after 7 days of incubation and added to the fermentation medium ( B) Homogenize to obtain 120 seeds in 50 ml aliquots and add this to the above Incubated for 8 days as in parts (c)(+) and (d)(+). 8 Fermentation broth (approximately 6 L) from flasks incubated for 1 day was filtered to remove bacteria. The thread body was removed. The filtrate was adjusted to pH 2.8 with sulfuric acid (20% v/v) and diluted with ethyl acetate. and back-extracted into sodium bicarbonate as described in part (a) above. Re-extracted into ethyl acetate at pH 2.8. Concentrate the ethyl acetate extract under reduced pressure. Condensation gave a yellow oil which was dissolved in methanol (10ml). This solution Evaporate to 31 and OD S -3 (Whata+an Partisil B ioprep 40.75, slurry filled with acetonitrile-water 20:80 -) column (32 x 2.5 cm).

The column had acetonitrile ratios of 1:4.3 and 2:3.1:1.3:2. eluted with a stepwise gradient of a mixture of acetonitrile and water, increasing to . Fractions were monitored by HPLC and fractions containing the title compound were evaporated and aliquoted. Setonitrile was removed. The resulting aqueous suspensions were pooled, lyophilized overnight, and the ash The title compound (59'+ng) was obtained as a white solid.

(e) Followed the procedure in part (d)(i) except that the pooled seed flasks were % (V/V) to inoculate 4 L of seed medium (A) in a 7B fermentor. culture At 500 rpm with agitation as above while aerating the culture at 4 L/sin. Incubated for 2 days. Remove 1.2L of culture and fill with 40L of seed medium (A). It was used to inoculate a 70L fermentor. The culture was 4QL/sin The culture was incubated as above for 2 days at 500 rpm with aeration of the culture. . Remove 15L of culture and place in 780L fermentor filled with 500L of fermentation medium (C) added to.

Fermentation medium (C): Fructose 50g Soybean oil 30g Cottonseed flour (Sigma) 20g neutral pH Culture was incubated for 450 hours at 200 rpm while aerating the culture at 500 L/1n. Incubate under shaking as above and add a 50% (V/V) solution of fructose. was increased to 5 g/L/day at 120 hours and 7.5 g/L/day at 162 hours. was supplied. Analysis of the broth at 450 hours yielded a title of 1056a+g/L. The yield of the compound is shown.

The above operation was repeated on a small scale, but fructose was replaced with glucose, galactose, Sucrose, maltose, lactose, myo-inositol, D-mannitol and other carbon sources selected from soybean oil. Each experiment at the 450th hour? Analysis of their blanks showed substantial potency of the title compound.

The title compound prepared according to the above procedure corresponded to a product with the following characteristics: Roughly 1690 molecules. -FAB mass spectrum 1lPJ constant [H]''''689. 2789; +F A B mass spectrum measurement [M+Na] 1713.27 53, molecular formula C35H46014 deuterium-500 MHz in chloroform ton N, +a, r, spectrum [multiplicity, coupling constant (H) and δ value with integral value]: 0.79-0.85 (m, 9H), 1. (10 (d, 7.3H), 1.04-1.15 (m, 2H), 2.09 (s, 310, 2.40 (m, 1) I), 2.69 (dd, 13.5. IH), 4.05 (s, IN), 4.94 (s, IIri), 4.98 (s, III) , 5.06 (d, 4. LH), 5.30 (s, LH), 5.78 ( d, 1B, 1. H), 5.92 (s, [1), 6.88 (dd, i s, g, [1), 7. l I (d, 7.2) 1), 7.14 (t, 7.IH), 7.24 (t, 7°2H).

Combined pulse decoupling 125.75MHz in deuterium-chloroform Carbon-13 N, i, r, spectrum [δ value showing the number of bonded protons in parentheses) : 171.5(0), 171.0(0).

189.1 (0), 167.0 (0), 16B, 7 (0), 1 57.9(1), 145.4(0), 140.1(f)), 12g, 9 (]), 128.1 (1), 125.8 (+), 1tL8 (1), H t, 4(2)・105.8(0), 118.5(0), 81.6(1 ), 80.7 (1), 79.3 (1), 75.1 (1), 74.2 ( 0), 42.9 (2), 39.7 (2), 36.7 (1), 34 ．． 2 (1), 33.6 (2), 31.6 (1), 29.4 (2), 25 ．． 4 (2), 20.9 (3), 19.8 (3), 18.8 (3 ), 13.5 (3), 10.9 (3).

Intermediate 2 A solution of the lyophilized product of Intermediate 1 (940 mg) in methanol (15 ml) was diluted with Treated with diazomethane (solution D (516 ml 0.4M) in methyl ether. Excess The diazomethane of was inactivated with acetic acid (0.1 ml) and the solution was concentrated under reduced pressure. was subjected to chromatography on silica gel (Merck 7734.50g) and Elute from chloromethane increasing to 2% methanol/dichloromethane, title Compound (906B) was obtained; "jo" N, n, r, (CDCl2) Te is δ0, 8-0, 9 (a+, Cdan 3), 1.04 (d, J-7H2 , CH-CHCHCf13), 2.09 (s, CH3Co2), 3.7G, 3 ．． 81 Oyobi 3.93 (3s, Co 2CJL3).

4,05 (d=J-2Hz, 7-H), 4.98 oyo (J' 5.00 ( 2s, C-CH2), 5, lO(d, J-6H2, CHOAC), 5 ．． 28 (S, 3-H), 5.75 (d, J-16H2, CH-C and c.

2), 5.81 (d, J=2) 1z, 8-H), 6-84 (dd, J"16 and 8.5Hz, C) I-CHCO2), 7.13-7.28 (1 , aromatic).

Intermediate 3 (IS-(la (4R*, 53*), 3a, 4β, 5α.

6α(2E, 4R, 6R*), 7β)Ill-(4-A* cetyloxy-5-methyl-3-methylene-6-phenylhexyl)-4,6, 7-Dolihydroxy-2,8-dioxabicycloC3,2,1)octane-3, 4,5-tricarboxylic acid, 6-(4,6-dimethyl-2-octenoate), 4.5-Dimethylester Intermediate 2 (173B) in tetrahydrofuran (51) A solution of aqueous sodium hydroxide (0, IN) was prepared at room temperature.

2.361). After 10 minutes, remove most of the organic solvent under vacuum. Ta. The resulting aqueous solution was diluted with water (201) and washed with ether (2x). water The solution was acidified with 0.5M aqueous hydrochloric acid, extracted with ethyl acetate (3x), and extracted with magnesium sulfate. It was dried with sodium hydroxide and filtered. Removal of solvent yields the title compound as a light brown gum. (145.2 mg) was obtained: Proton N, m, r, (CDCla) is 67 ．． 1-7.3 (m, 5H-Ph), 6.85 (dd, H(, CH-CHCO2 , J-15,5 and 8.7511z), 5. lH(d, lit, C ll0COCH-CI, J-211z), 5, 75 (d, il l　,　e■Soot|Ct(C0 2, J-1-5,5Hz), 5.22 (s, lH, C Dan CO2H), 5.0[1 (d, l11.Ac0cl(, J-5+1z), 5, 01 (s, 2 H, C-CB2), 5.0 (br s, LH, ON), 4.0e (d, 1N.

CHO) (, J-21 (Z), 3.'13 and 3.79 (2S, 8H12C0 2Me), 2.89 (dd.

LH, PhC) I (7) 1, J-13.7 Oyobi fi, 2H2), 2.1 (S, 314.Cu3Co, ), 1.05 (d, 3) 1. CB-CHCII C tempa, J-7Hz), 0.8-0.9 (m, 9tl, 3C dan,).

Intermediate 4 cetyloxy-5-methyl-3-methylene-6-phenylhexyl)-3-(a (minocarbonyl)-4,6,7-dolihydroxy-2,8-dioxabicyclo[ 3,2゜1] Octane-4,5-dicarboxylic acid, 6-(4,6-dimethyl-2 -octenoate), 4,5-dimethyl ester Dry N,N-dimethylene in dry dichloromethane (151) cooled to 0 °C under nitrogen. Oxalyl chloride (1,1 1) was added. The mixture was stirred at 0°C for 5 minutes and then dissolved in dry tetrahydrofuran ( 22ml) and the intermediate (3g) in dry acetonitrile (13ml) were added.

The mixture was stirred for 2 hours at 0°C under nitrogen, during which time gaseous ammonia was added for 0.5 hours. He blew gently. After heating to 20℃ and leaving for 2 hours, water (20ml) was added and extracted with ether (40 ml). After washing with water, dry the extract, Evaporation gave the crude product, which was purified by silica gel (Merek 9385.600). ml) chromatography eluting with ethyl acetate dipetroleum ether (5:1) Purification to give the title compound (1.75 g) as a pale foam: Proton N , Lr, (CDCI), δ0.72-0.90(m, 911.CB, ), 0.99(d, 3H, J-7, 5Hz, -CHCHCI(3), 2.10( s, 3H, 0COCdan 3), 3.34 (s, LH, 7-0) 1), 3.82 (s , 3H, Co CH), 3.95 (s, 3H, Co2C and 3), 4.05 (s, LH, 7 once), 4.99 and 5. Ql (2s, 2H, -CB, ), 5.0 9(m, 2)1, CH-OAc and 3-H), 5.55(brs, l) l, NH), 5.76 (d, LH.

J-15) 1z, C1 (-C8-Co), 5.80 (s, LH, 6-H), 6 ．． 50 (br　s, LH, Nji), 13.85 (dd, IH, J-15, 7H z, CH-CH-Co), 7.10-7.32 (country, aromatic proton) .

Analysis actual value: C61,34:H7,04, N 1.94CHNo, 0.5H 20 calculated value d A5113 CB1.08. H7, L5, N 1.92% Muyum"hi"" Utsuyuemu 1 earth L-mu" Yukoel upper yu (" Yoeyu mutual two and 1" 2 (4,6 nimuri 22" Yu L mmum) Intermediate 4 (2,786 g) was dissolved in dichloromethane. Dissolved in methane (83 ml) and triethylamine (1,59 ml), Trifluoroacetic acid (1.09ml) was added. The solution was stirred at room temperature for 3 hours and then Between triethylamine (0.4 ml) and trifluoroacetic anhydride (0.27 ml) Add 1).

The solution was stirred for a further 2 hours at room temperature, diluted with dichloromethane (100ml) and diluted with 2N Wash with hydrochloric acid (2001) and aqueous saturated sodium bicarbonate solution (200a+I). Purified. The organic phase was dried (MgSO4) and evaporated to give a tan gum. child Silica gel (Merck Kiesclgel 60.100g, 240~ 400 mesh) chromatography using cyclohexane:ethyl acetate (3 : It was eluted with 1). Combine the appropriate fractions and evaporate the title as a colorless foam. Compound (2.30 g) was obtained; proton N, O1゜r, δ in (CDC13) 7.33-7.10 (m, aromatic proton), 6.85 (dd, J-16, 8Hz, OCOCH-CHCHMe), 5.74 (d, J”l [i Hz, 0COCJi”CHCHMe), 5.66(d, J=2.5Hz, H-6), 5.50 (s, H-3), 5.07 (d, J-5Hz .

CHCo CH), 5.0 and 4.95 (2s, -CH-2), 4.05 (s , 4-0H). 4.03 (d, J-2, 5Hz, H-7), 3.9[i and 3 ．． 81 (2s, Cdan 302C), 3.26 (d, J-2,5Hz, 7-0H), 2.68 (dd, J-14Hz, 5Hz, 1 of Chi2Ph), 2.09 (s, C and Co 2-), 1.04 (d, J-6) 12,0 COCH-CH CHC Dan. ) is shown.

Analysis actual value: C63, 52, H7, 27, N 2.09 C37H4, NO12 Calculated value: C63,50, H7, O[i, N 2.00% cetyloxy-5 -Methyl-3-methylene-6-phenylhexyl'I-4,6,7-1-hydride Roxy 2,8-dioxabicyclo[3,2,1]octane-3,4,5-tri Carboxylic acid, tripotassium salt A suspension of the lyophilized product (1 g) of intermediate 1 in water (100 nl) was added to water (10 ml). Treated with a solution of potassium bicarbonate (430 mg) in water. Lyophilize the resulting solution The title compound (1.08 g) was obtained as a beige solid; ol) 3491-3167 (br OH), 1731 (ester C-0) 1 aX , 1614 cm-' C carboxylate co and C-C); Analysis actual measurement Value 2 C48,65; H5,70; K 14.1 i H20B, 2C35H4 3 to 3o14・3H20 calculated value: C48,93; H5,75; K 13.65 ;H20G, 29% intermediate 7 (1s-(1α(4R,5S*), 3α, 4β, 5α.

* A stirred suspension of intermediate 6 (15.6 g) in methanol (IL) was dissolved in concentrated hydrochloric acid (13 m 1) was added dropwise. The resulting clear solution was stirred at room temperature for 24 hours. Then that This was treated with solid sodium bicarbonate (13.2 g) and most of the solvent was removed under reduced pressure. Evaporated. The residue was treated with aqueous hydrochloric acid (2M, 500ml) and ethyl acetate (I LX3). The organic extract was washed with water (IL) and dried over magnesium sulfate. Dry, filter and evaporate. The residue was dissolved in dry toluene (130+al) and nitrogen Heating to 80°C under It was treated dropwise with tar (38 ml) over 30 minutes. The reaction mixture was heated to 80°C for 31 min. The mixture was stirred for /4 hours and then cooled. Dilute it with ether (700 nl) and salt Washed with water (600 nl). Dry the organic layer over magnesium sulfate and remove the solvent under reduced pressure. evaporated under. The residue was immersed in silica gel (Merck 9385.900 g) 5:1 to 1:1 cyclohexane:vinegar Elution was carried out with ethyl acid. Combine the appropriate fractions and evaporate the solvent to a yellow foam. The title compound (5.93 g) was obtained; proton N, m.

r, (CDCI) value is 60.8-0.9 (i, 9H13Cdana), 1.0 5(d,3)1. -CHCHCdana, J-6, 2Hz), 1, 8 and 1 ．． 48 (2S, 18H, 2CO2C (C and 3) a), 2. l(s, 3H, Cdan3　CO□　), 2.71 (dd, 1) 1. One of PhC) I, J-13 , 7 and 5Hz), 2.96 (d, LH, CHOH, J-2Hz), 3.73 (s, 3H.

C02Cdana), 4.1 (s, LH, OH), 4.05 (t, 18.C and 0) 1. J-2Hz), 4.97 (s, 2H, C-CH2), 5.1 (d, t)1 ．． ^cOcH, J-5Hz>, 5.28 (s, LH, cHCo CH), 5.7 7 (d, IHoCH-CHCO2, J-16, 2Hz), 5.97 (d.

1H,Cdan0COC)I-CH,J-2)1z),6.91 (dd,IH,C ) I-CHCO2, J-16,2 and 8.7) 1z), 7.1-74 (11, 5H, P! L); t, 1. c, (SiO,,)ethyl acetate/cyclohexane (1:1) Rfo, 5B Intermediate 8 (1s-(1α(4R,5S'), 3α, 4β, 5α.

Book cetyloxy-5-methyl-3-methylene-6-phenylhexyl)-7-(( Dimethyl(1,1-dimethylethyl)silyloxy)-4,6-cyhydroxy -2゜8-dioxabicyclo(3゜2.1)octane-3,4゜5-tricarbo acid, 6-(4,6-dimethyl-2-octenoate), 4,5-bis(1, 1-dimethylethyl) ester, 3-methyl ester Intermediate 7 (10,38 g) in dry dimethylformamide (26 ml), t-butyl of dimethylsilyl chloride (19,6 g) and imidazole (17,7 g). The solution was stirred at 65°C under nitrogen for 16.5 hours, then ethyl acetate (200+nl) and 2M hydrochloric acid (200nl).

The aqueous phase was extracted with ethyl acetate (2001), the combined organic extracts were extracted with 2M hydrochloric acid (1 00 nl), washed with water and brine (2 x 100 ml each) and dried (MgS04). Dry and evaporate to an orange oil. Add this to silica gel (Merek 77) 34.500g) and chromatographed with 4:1 cyclohexane:acetic acid. It was eluted with chill. Combine the required fractions, evaporate and title as a colorless gum. A compound (9.01 g) was obtained: Proton N, II1. r, (CDCI3) is 6 0.05(s, (Cdan, )2St), 0.8-0.9(a, CH3 and (C Dan. ) a C3i), 1.02 (d, JJHz, -CHCHCHa), 1. 4 and 1. B5 (2s.

Co　C(CI　　)　), 2.1(s, CHCO), 3.73(S, co2c ! L3), 4°02 (s, 4-inch), 4.12 (d, J-2Hz, 7-H), 4 ．． 98 and 5.0 (2s, C-CH2), 5.12 (d, J-5Hz, C-CH2) 0Ac), 5.28 (s, 3-H), 5.8 (d, J-IBHz, 0COC ) 1-CH), 6.38 (d, J-2Hz, 6 once), 6.93 (dd, J-9 and 16Hz, 0COCH=CI), 7.1-7.3(m, C6dan5). vinegar.

Intermediate 9 (Is-(1α (4R, 5g), 3α, 4β, 5α.

Book cetyloxy-5-methyl-3-methylene-6-phenylhexyl)-7-[[ Dimethyl(1,1-dimethylethyl)silyloxy)-4,6-cyhydroxy -2゜8-dioxabicyclo(3,2,11 octane-3,4゜5-tricarbo acid, 6-(4,6-dimethyl-2-octenoate), 4,5-bis(1, 1-dimethylethyl) ester A solution of intermediate 8 (9, OIg) in tetrahydrofuran (450 nl) was stirred at room temperature. Under stirring 0. Treated with IM sodium hydroxide (116 nl). Solution after 0.5 hours was evaporated to a small amount, then ethyl acetate (250+al) and 2M hydrochloric acid (500n l). The aqueous phase was further extracted with ethyl acetate (2×250+nl). Match The combined extract was washed with water and brine (2 x 250 ml each) and dried (MgS04). Drying and evaporation gave the title compound (8.7g) as a white foam; proton N , I1. r, (CDCIs) is 60.04 (s, (Cl3)2St), 0 ．． 8-0.9 (1, CH, and (C, j, )3C3i), 1.02 (d, J-6Hz, -CHCHCHa), 1.4 and 1.85 (2s, Co2C( Cp3) 3).

2.1 (s, CHa Co), 4.13 (d, J-2Hz, 7-Jj), 5.0 2 and 5.05 (2s.

C-CH,), 5. Ll(d, J-5)1z, CHOAc), 5.22(s, 3 once), 5.78(dl-16Hz, 0COCI("CH), t3.34( d, J-2Hz, G once), 6.95 (dd, J-9 and 16Hz, 0C OCH-Cl), 7.1-7.3 (country, C6dan5).

Intermediate] 0 * (Is-C1α (4R, 5S books), 3α, 4β, 5α.

Intermediate 9 (5,89 g) in tetrahydrofuran (60 ml), N-hydroxy Cysuccinimide (0,8g) and N,N-dicyclohexylcarbodiimide A solution of (1.46 g) was stirred at room temperature for 17 hours. The resulting suspension was filtered; The filtrate was evaporated to a white foam.

Dissolve this in dimethylformamide (60111), stir at room temperature, and Treated with sodium urethane (242 mg). After 55 minutes the suspension was filtered. filter The solution was partitioned between ethyl acetate (500 ml) and 2M hydrochloric acid (500 ml). organic phase water (500ml), saturated aqueous sodium bicarbonate and brine (2X500ml each) 1), dried (MgSO4) and evaporated. The residue is treated with silica (Merck 7734.300 g) and 3:1 cyclohexane: Elution was performed with ethyl acetate. The required fractions were combined, evaporated and marked as a white foam. The title compound (2.55 g) was obtained; Bouton N, m, r, (CDC1a) Teha 60.04 (S, (C) 13) 231), 0.8-0.95 (w, C, j, and (Ci3) 3CSi), 1.02 (d.

JJHz, -CHCHC 3), 1.4 and 1.131 (2S, Co2C (C H, > 3), 2.1 (s, Cl, Co), 3.58 and 3.78 (CH 20H), 3.86 (S, 4-0H), 4.11 (d, J-2Hz, 7-H) , 4.65 (dd, J-4 and 6Hz, 3-)j), 4.98 and 5.0 (2S, C-CH2), 5.12 (d, J-5Hz, 0 Ac on q), 5.8 (d , J-1[iHz, 0COCdan-CH), 6.32 (d, J-2Hz, B-H) , 13.92 (dd, J-9 and 18Hz, 0COCR-C) I), 7.1- 7.3 (+, C6H5).

Intermediate 11 Zero * (Is-[1α(4R,5S), 3α, 4β, 5α.

Kutaji 4,5-dicarboxylic acid, 6-(4,6-dimethydichloromethane (31 ) of dimethyl sulfoxide (0.4 ml) was stirred at -70°C under nitrogen and diluted with dimethyl sulfoxide. 1 with a solution of trifluoroacetic anhydride (0°6 ml) in chloromethane (1,5 ml). The dropwise treatment was carried out over a period of 2 minutes. After a further 10 minutes, in dichloromethane (1.5 ml) A solution of Intermediate 10 (495a+g) was added dropwise over 15 minutes. Stir about -70 ℃ for 30 minutes, then add triethylamine (0°9 ml) for about 15 minutes. dripped. The reaction mixture was allowed to warm to room temperature over approximately 30 minutes, then ethyl acetate (5 01) and water (50Ql+). The organic phase was dissolved in water (50 ml) and brine (2 Washed with X50a+1), dried (MgSO4) and evaporated to a yellow gum. I let it happen. Chromatography of this on silica (Merck 7734.30g) and eluted with 5.1 and then 4:1 chlorohexane:ethyl acetate. necessary Fractions were combined and evaporated to give the title compound (315B) as a pale yellow gum. ;　ν(CHBra　)ax 1728 and 1739 cm-1; for protons N, m, r, (CDCl5) 60.03-0.6 (2s, (CH) S+), 0.8-0.95 (m, CH a and (CI) C3i), 1.02 (d, J-6Hz, -〇HCHCdan s), t, 4 and l.

65 (2s, Co C(CH)), 2.1 (s, Cdan a Co), 4, 0 (s54-01), 4°2), 5.14 (dl-5Hz, CHOAc), 5.80 (d, J-16Hz, 0COCH-C1l), 6°28 (d, J-2H z, 13-H), [i, 93 (dc!, J-9 and I GHz, 0 COC I(-C1(), 7-1-7.3(m, C6H5), 9.5(Cl0) vinegar.

Intermediate 12 * [:IS-[1α(4R,5S”), 3α(EZ).

1-(4-acetyloxy-5-methyl-3-methylene-6-phenylhexy )-7-((dimethyl(1,1-dimethylethyl)]silyloxy)-4,6 -cyhydroxy-3- [(hydroxyimino)methyl)-2,8-dioxabi Cyclo[3,,2,1]octane-4,5-dicarboxylic acid, 6-(14,6- dimethyl-2-octenoate), 4,5-bis(1,1-dimethylethyl)ethyl Stell A solution of intermediate 11 (244 o + g) in pyridine (2.5 m) was stirred at room temperature, and the salt Treated with acid hydroxylamine (24111 g). After 2 hours, ethyl acetate (5 01) and the solution was mixed with 2M hydrochloric acid (2X 25ml), water (2X 25ml) and Washed with brine (2 x 25 ml), dried (MgSO4), evaporated and freed. The title compound (250 mg) was obtained as a colored gum: protons N, m, r, ( CDCIs), δ0.05(2s, (CHs)2Si), 0.75-1 ．． 0 (rA, CH3 and (Cfi, )3C3t), 1.02 (d, J-6H z, -CHCHCdan3), 1.4 and 1y61 (2s.

Co,, C(CH,) 3), 2.1(s, Cdan a Co), 3.92(s , 4-0. jj), 4-12 (d.

J=2Hz, 7-)1), 4.95 and 5.0 (2s, C-CH2), 5. 12 (d, J=5Hz, CHOAc), 5.18 (d, J-7Hz, 3-H), 5.81 (d, J-16Hz, 0COCH-CI).

8.37 (d, J-2Hz, 8-H), 8.95 (dd, J-9 and 18H z, 0cOcH-CI), 7.1-7.3 (m, C6dan5), 7.38 (d, 7 )1z, cH-N).

Analysis actual value: C64,5; H8,8,N 1.2C49H77N Otas l Calculated value: C84,2, H&,5;N 1.5%* 6α (2E, 4R, 6R’), 7β)) 1-(4-aviridin (2,5+nl) A solution of middle intermediate 12 (226 mg) was stirred at room temperature and diluted with trifluoroacetic anhydride (7 0 μL). After 55 minutes, ethyl acetate (50a+I) was added and the solution was diluted with 2 Washed with M hydrochloric acid, water and brine (2X25a+l each) and dried (MgS04). , evaporated to give the title compound (223 mg) as a colorless gum: Proton N , I1. r.

(CDCI) is 60.05-0.08 (2s, (Cdana) 2S+) , 0.8-1.0 (m, CI! and (Ctl) C3+), 1.02 (d , J-[1)1z, -C)Ic)IC,jja), 1.4 and 1. [15( 2s, COC (CH)), 2.08 (s, C and 3CO), 4.12 (d, J- 2Hz, 7-,! ri, 4.96 and 5.02 (s, cc and 2), 5.1 1 (d, J=5Hz, CHOAc), 5.55 (s, 3-1j), 5.79 (d , J-16Hz, 0COCH-CH), 8.22 (d, J-2H2,6-H), 8.95 (dd, J-9 and 16Hz, 0COCR-CH), 7.1-74 ( ■,C6dan,) is shown.

Analysis actual value: C65.0, H8.6; N 1.2C49H75N OL2S l Calculated value: C65.5, H8.4; N 1.8%6α (2E, 4R, 6H pieces) ,7β:1)1-(4-a* A solution of intermediate 13 (207 g) in tetrahydrofuran (5 ml) was stirred at room temperature. Stir and add tetra-n-butylammonium fluoride (IM in tetrahydrofuran). solution, 0.231). After 65 minutes the solution was evaporated to dryness.

The residue was partitioned between ethyl acetate (50ml) and water (50ml).

The organic phase was washed with water (50ml) and brine (2X50ml), (MgSO4) Drying and evaporation gave the title compound (193mg) as a gum; proton N, I1. r, (CDCI3) is 60.8-0.95 (m, CM), 1 ．． 05(d, J”6Hz, =CHCICdan.

), 1.48 and 1.58 (2S, CO2C (CH3) 3), 2.10 (s , CM, c.

), 4.05 (+a, 7-H), 4.95 and 4.98 (2S, C-Cl+ 2), 5.08 (d, J=5, a2(a, C6dan5).

Mass spectrum (MW 783.9) + FAB 784.4288 (M)I) cetyloxy-5-methyl-3-methylene-6-phenylhexyl)-4,6, 7-Dolihydroxy-3-(IH-tetrazol-5-yl)-2,8-dioxy Sabicyclo[3,2,11 octane-4,5-dicarboxylic acid.

6-(46-dimethyl-2-octenoate), 4°5-dimethyl ester Intermediate 5 (1 g) was dissolved in N,N-dimethylformamide (401). Horse mackerel Add sodium chloride (650 mg) and triethylamine hydrochloride (1,375 g). Then, the mixture was stirred at 120° C. for 2 hours under nitrogen. Cool the mixture to room temperature and add nitrous acidified with 2N hydrochloric acid, and diluted with ethyl acetate (2X 150 1). The organic extracts were combined, washed with water (4X 100ml), and (M gSO), dried and evaporated to give a yellow oil.

This was dissolved in ether (50at) and washed with water (3X3(1+1). The phase was dried (M g S O4) and evaporated to give a colorless foam. Prepare this lative HPLC [5pherisorb 0DS-2 column, flow rate 15all lins 66% (95:5 acetonitrile:water and concentrated sulfuric acid 0.15ml/L ) and 3496C water and concentrated sulfuric acid 0.15 ml/L)). appropriate The fractions were combined and the acetonitrile was removed under reduced pressure at 40°C. acetic acid to remove the cloudy aqueous phase. Extracted with ethyl (2X2001). Combine the organic extracts (Mg 504) Drying and evaporation gave the title compound (93B) as a colorless solid; proton N , w, r, (DMSO-d6) has δ7.30-7.11 (m, aromatic proto ), 8.92 (dd, J-17, 5Hz, 8.7511z, OCOCH-CH CHMe), e, 41 (d, J-5Hz, 7-Off), 8.12 (d, Jc2 11z, H-6), 6.21-6.13 (bs, Nll or 4-0H), 5 ．． 98 (s, 1l-3), 5.82 (d, J-17, 5Hz, OCOC) 1- CHCHMe), 5.02(d, J-4)1z, CHOCOCH3), 4. ’ 16 and 4.92 (2s, = C) 12), 4.05 (dd, J-5Hz, = 2Hz, H-7), 3.73 and 3.80 (2s, Co C8), 2.64 ( dd, J=12.5Hz, 6Hz, (1 piece of 42ph), 2.10(s, 0 COCjj3), 1.0 (d, J=7Hz, 0COC)l-CHC) ICC20 shows.

Analysis actual value: C59.12; H6.8g, N 7.27Ca7HsoN 4012.0.5H20 calculated value: C59, 11, Hfli, 84, N 7.4 5% example 2 (IS-[1α(4R,5'S*), 3α, 4β, 5α.

* * 6α(2E,4R,6R”),7β))1-(4-A The title compound of Example 1 (3,5 53g) was dissolved in 2.4.6-collidine (130QI+). lithium iodide (4.88 g) was added and the mixture was stirred at 45° C. under nitrogen for 24 hours. Pour the mixture into the chamber. Cool to warm, pour into 3N hydrochloric acid (1000ml) and add ethyl acetate (2X 500ml). n+l). The combined organic extracts were diluted with 3N hydrochloric acid (2X2501) and water (2 501), saturated aqueous sodium thiosulfate solution (250 ml), and water (250 ml). Washed and then dried (MgSO4). Evaporate the solvent to leave a brown foam. I obtained this and used it as a prevariety HP LC (Spherisorb 0DS-2 card). ram, flow rate 15 m1/min, 60% (95:5 acetonitrile:water and concentrated sulfur) acid 0.15+nl/L) and 40% (water and concentrated sulfuric acid 0°15Ill/L)) It was purified by Appropriate fractions were combined and the acetonitrile was evaporated under reduced pressure.

The cloudy aqueous phase was extracted with ethyl acetate (2X300ml). Combined organic extracts (MgS04) Dry and evaporate to give a beige solid. Some of this substance ( 860 mg) was dried under vacuum at 45°C for 24 hours as a beige solid. A compound (784 mg) was obtained. Boo ton N, s, r, (CD300) Teha 6 7.27-7.08 (m, aromatic proton), fi, Ilg (dd, J-1 [i Hz, 8) 1z.

0COCH-CHCHCH3), 6.44(d, J<2)1z, H-6), 6. 24(s,)I-3), 5.82(d, J-16Hz, 0COCH-CHC)I CH3), 5.06(d, J-5)1z, CHOAc), 5.02 and 4.9 8<2s, -Cdan2), 4.14(d, J<2Hz, H-7), 2.1li 7 (dd, J-14Hz, 7Hz, CH2Ph) 1), 2.10 (S, 0 COCH3), i, 04 (d, J-8Hz, 0COCH=CHCHCH3) show.

Analysis actual value: C5[i, 55:HG, 59; N7.39; H202, 8C35H46N4012””Ethyl acetate, 1.25H20 calculated value: C56, 1!5; He, go; N 7.09; H202, 85% failure [IS-(lα(4R*,5S”), 3a, 4β, 5α.

The title compound of Example 2 (500 mg) was dissolved in potassium bicarbonate (207 mg, 2.95 equivalents). (amount) suspended in water (140 ml). The resulting suspension is dissolved until it becomes a complete solution. Ultrasonicated. The solution was extracted with ether (100 ml), the aqueous phase was lyophilized and The title compound (511B) was obtained as a beige solid; protons N, m, r , (D20) is 67.37-7, L7 (m, aromatic proton), 6.9 7 (dd, no 16Hz, 8) 1z, OCOC)I-C)IC)IMe), 6 ．． 34 (d, J<2Hz, H-6), 5.97 (s, H-3), 5.94 (d, J-16t (z.

OCOCH-C)1cHMe), 5.04 and 4.97 (2s, -Cjj2) , 4.91(d, J-4)1z, CHOAc), 4.04(d, J<2Hz, H -7), 2.16 (s, 0 COCH3); HPLC retention time 10.14 m 1n (Spherisorb 0DS-2 column (20cIIIXO, 4cm ), flow rate 1, 5 ml/win, 6a% (95:5 acetonitrile:water and and concentrated sulfuric acid 0.15 m1/L) and 40% (water and concentrated sulfuric acid 0.15 mark +/L ))Ne * 6α (2E, 4R, 6R), 7β)) 1-(4-A* 6α(2E, 4R, 6R*), 7β)) 1-(4-acarboxylic acid, 6-(4, 6-dimethyl-2-octenoe), 4,5-dimethyl ester (B) Example 1 (0.4 g) dissolved in N,N-dimethylformamide (5 ml) in 1° carbonated water Add base sodium (0.48g) and iodomethane (0.241g) to the mixture. was stirred for 16 hours. The mixture was then poured into brine (50 ml) and ethyl acetate ( 100ffll). The organic layer was washed with water (3X50ml), dried and Evaporated. The residue was soaked in silica gel (Merck Kieselgel l1iO, 100gs 230~400 mesh) Elution was with xane:ethyl acetate 3:1 to 1:1. Appropriate fast running minutes The fractions were combined and evaporated to give the title compound A (0,153 g) as a white foam. Proton N, m, r, (CDCIa) Te4;! δ7.30-7.06 (m, aromatic proton), 6.88 (dd, J-16 and 8Hz, 0COC H-CHCHCH3), [i, 1g (s, H-3), 5.87 (d, J-2, 5Hz, )I-6), 5, 78 (d, J-16Hz, 0COCR-CHCjI CH), 5.05 (d, J-5Hz, CH3C02CH), 5.00 and 4 ．． 95 (2s.

-CH2)-4,18 (s, NCHa), 4-14 (m, H-7), 3. 98 and 3.84 (2s, c02C) 13), 3.32 (d, J-2,5Hz , 7-0H), 2.68 (dd, J-14 and 5Hz, LH, Cdan 2 Ph) , 2.08 (s, Cdan a CO2), 1, 05 (d, J-6Hz.

0COCH-CHCHCH3).

Analysis actual value 2C[io, 52:H7,24; N 7.12C38H52N40 12 Calculated value: C [io, 30; H6, 93; N 7.40% appropriate slower The running fractions were combined and evaporated to give the title compound B (0,0 77g) was obtained; proton N, w, r, (CDCIa) was 67.30-7 ．． 08 (m, M aromatic proton), 6.88 (dd, J-16 and 8Hz, 0C OCH-CHCHCH3), 8.08 (s, H-3), 5.9 [i (d) , J-2,5Hz, H-8), 5.78 (d, J”18Hz, 0 COCH=CHCHCH), 5.1O (d, J”5Hz, CH3C02CH) , 4.98 and 4.97 (2s, -CH), 4.30 (s, NCdana), 4.14 (br　s, H-17), 3.88 and 3.82 (2S, CO2C 3), 3.35 (d, J-2, 5Hz, 7-0R), 2.70 (dd,)-1 4 and 5) IZ, LH, c) I, Ph).

Analysis actual value + CBo, 46; H7, 1B, N 7.29C38H52N4 012 calculated value: Ceo, ao;) IB, 93, N 7.29% Cecilio xy-5-methyl-3-methylene-6-phenylhexyl)-4,6,7-doly Hydroxy-3-(1-methyl-IH-tetrazol-5-yl)-2,8-di Oxabicyclo(3,2,1)octane-4,5-dicarboxylic acid, 6-(4, 6-dimethyl-2-octenoate) Example 4 Compound A (357 mg) was dissolved in 2.4.6-collidine (20 ml) . Anhydrous lithium iodide (548 mg) was added and the mixture was heated at 40°C under nitrogen for 16 hours. Stirred. The mixture was cooled to room temperature and treated with 2N hydrochloric acid (200ml). this was extracted with ethyl acetate (200ml), and the organic phase was further extracted with 2N hydrochloric acid (2X 50ml). ) Washed with water (50 ml). Dry the solution (N a 2 S O4) , and evaporated to give a dark gum. This is preparative HP L CC5ph erisorb 0DS-2 (25X 2c+Il) column, flow rate 15 ml /sin.

68% (95:5 acetonitrile:water containing concentrated sulfuric acid 0.15ml/L) and and 32% (water containing 0.15 ml/L of concentrated sulfuric acid)]. appropriate amount The fractions were combined and the acetonitrile was evaporated under reduced pressure. Salt water (50ml) became cloudy Added to the aqueous phase and extracted the product with ethyl acetate (3X1001). Combine the organic extracts dried (Na2S04) and evaporated to give the title compound as an off-white solid foam. (104II1g) Obtained; F0 tons N, m, r, (CD30D)” C'ha δ7.2B-7,0B (11, aromatic proton), 8.1111 (dd, J-16 and l1Hz.

0COCH-CdanCHCH3), 6.48(d, J=2)1z, H-8), 6. 30(s,)l-3), 5.84(d, J-16Hz, 0COCH-CHC)1 cH3), 5.04 (d, J-5Hz, CHOAc), 5.02 and 4.98 (2s, C=C)12), 4.24(s, NCjj3　), 4.12(d, J- 2Hz, H-7), 2.65 (dd, J-14 and 8Hz, IH, Cdan 2P h), 2.08 (s, cIIOc.

Cjia), 1.04 (d, J-7Hz, 0COCH-CHCHCdan3). vinegar. Exact mass measured 729.3309; MH+calculated 729.334 7* 6α (2E, 4R, 61ne), 7β)) 1-(4-armethyl-2H-tetra sol-5-yl)-2,8-dioxabicyclo(3,2,1)octane-4, 5-dicarboxylic acid, 6-(4,6-dimethyl-2-octenoate) Example 4, compound B (170 mg) was dissolved in 2.4.6-collidine (10 ml) . Anhydrous lithium iodide (309 mg) was added and the mixture was heated at 40°C under nitrogen for 40 hours. Stirred. The mixture was cooled to room temperature and treated with 2N hydrochloric acid (200a+l). child This was extracted with ethyl acetate (200nl), and the organic phase was further extracted with 2N hydrochloric acid (2X50+nl). l), aqueous sodium thiosulfate (10%, 50!1Il) and water (50ml) Washed with. Dry the organic solution (Na2S04) and evaporate to a dark gum. I let it happen. This is preparative HP L C (Spherisorb 0DS- 2 (25X 2 cm) column, 9 containing 65% (concentrated sulfuric acid 0.15 ml/L) 5:5 acetonitrile:water) and 35% (concentrated sulfuric acid 0.15a+I/L) purified water)]. Combine appropriate fractions and evaporate acetonitrile under reduced pressure. I let it happen. Brine (50+al) was added to the cloudy aqueous phase and the product was extracted with ethyl acetate. Ta. The organic extracts were combined, dried and evaporated to give the title compound ( 89 mg) was obtained; proton N, m, r, (CDa OD) was 67.30-7 ．． 08 (m, aromatic proton), 6.88 (dd, J-16 and 8Hz.

0COC) 1-C) 1cHc) 13), 6.46 (d, J-2112, 1l-G ), 6.31 (S, I+-3), 5.1114 (d, J-IBllz, 0COC 1j-CHCIIC113), 5.08(d, J-5112, CpOAc), 5 ．． 02 and 4.9 [1 (2s, C-CH2), 4.33 (s, NCdana), 4.10 (d, J-2Hz, H-7), 2. H(dd, J-14 and GHz, IH, cp2Ph), 2.09 (s, C)IOCOCH,), 104 (d, J= 7Hz, 0COCH-CHCHCH,').

Analysis actual value: C57,08:H6,79; N 7.03036H48N40 12.1.5H20 calculated value: C57,21,H6,80; N 7.41%* 6α (2E, 4R, 6R units), 7β)) 1-(4-A The title compound of Example 5 (97m g) in ether (20 ml), water (20 ml) and potassium hydrogen chloride (2 , 6tag). Separate the aqueous layer and wash with ether (2X20ml). and then lyophilized to give the title compound (97 mg) as a white lyophilized solid. Obtained. Proton N, a+, r, (D20) is 67.36-7.14 (n, aromatic proton), 6.98 (dd, J-16C-C12), 4.90 (d, J =3Hz, CH3Co2Cdan-), 4.22(s, NCdana L4.09(b r　s,)l-7), 2.16(s-CHs　Co-).

Analysis actual value: C50, 32; H5, 82, N 8.0gc36H46N4 2 012.3H20 calculated value: C50,33; H6,10, N Ili, 52% Example 6 The title compound (62 mg) was dissolved in dioxane (5+al) I, and water (10 ml) was dissolved in dioxane (5+al) I. ) in hydrogen carbonate (17 mg). Lyophilize the resulting solution overnight The title compound (64 mg) was obtained; protons N, m, r, (D20) It's coming δ7.40-7. l[1(rA, aromatic proton), 8.98(dd, J -16 and 8 Hz.

0COC11-Cdanc11cH3), 8.29 (s, H-6), 6.00 (s, H-3), 5.95 (d, J-IBHz, 0COCH-CtlCHCH), 5. 06 and 4.98 (2s, C-C dan 2), 4.9G (d, J-5Hz, -C HoAc), 4.3tli (s, NC, i 3), 4.06 (s , )I-7)-2,18(s, CH3CO).

Analysis actual value: C49,04:H5,60; N 6.02C49,30:H6 , 21; N Ili, 39%-5-yl)-2,8-dioxabicyclo( 3,2゜dimethylformamide (51) middle side 4, compound A (756Il1g) N-methylhydroxylamine hydrochloride (261 mg) and triethylamine ( 0,882g+1) for 16 hours. Dimethylformamide under reduced pressure Evaporated and the residue was partitioned between ethyl acetate (1501) and water (1001). The machine layer was washed with water (10C]+I) and brine (100+ol), then (Na2S 04) Dry and evaporate. The residue was soaked in silica gel (Merck Kieselge) 80.150 g, 240-400 mesh), Elution was performed with cyclohexane:ethyl acetate 3:1 to 1:4. Combine appropriate fractions , evaporated to give the title compound (413a+g) as a solid foam; proton N , m, r, (CDC13) is 67.38-7.06 (+a, aromatic proton ), 6.10(s, H-3)-5,22(dd, J-5,5 and lHz, H -8), 5.04 (d, J-5Hz, -CHoAc), 4.98 and 4.9 7 (2s, -Cdan, , ), 4.28 (dd, J-5 and IHz, H-7) , 4.22 (s, -NCH3), 4.08 (s, 4-0 degrees), 3.94 and 3 ．． 83 (2s, 2" CO2C Dan 3), 3.Ei4 (d, J-5t+z, 7 -0H), 3.30 (d, J-5,5H2,11i-OH), 2.68 (dd, J-13 and 6Hz, ill.

CHPh), 0.116 (d, J-tliHz, CHC Dan,).

Analysis actual value: C53,6, H6,03; N 8.4CHN O,1,5H2 0 calculated value: C53,24; HG, 22; N 8.87% 6α, 7β: 1) 1-(4-A cetyloxy-5-methyl-3-methylene-6-phenylhexyl)-4,6, 7-Dolihydroxy-3-(1-methyl-IH-tetrazol-5-yl)- 2,8-dioxabicycloC3,2゜1]octane-4,5-dicarboxylic acid, 4 - Methyl Nissa The title compound of Example 9 (401 mg) was dissolved in 2.4.6-collidine (15 ml). Ta. Anhydrous lithium iodide (887 mg) was added and the mixture was heated at 40°C under nitrogen for 32 hours. and then stirred at ambient temperature for 64 hours. Collidine was evaporated under reduced pressure and the residue Partitioned between brine (20ml) and ethyl acetate (100ml). The aqueous layer was further diluted with ethyl acetate. (2×100ml). Combine the organic layers, (NaSO4) Dry and evaporate to a gum. This is Prevarate HP L CC 3pherisorb 0DS-2 (2X 25cm) column, eluent 40% ( 95:5 acetonitrile:water + trifluoroacetic acid 0.1 + al/L) and 60 % (water + trifluoroacetic acid 0.11/L), flow rate 15 ml/akin) and purified.

Appropriate fractions were combined and the acetonitrile was evaporated. Lyophilize the aqueous residue and ash The title compound (29 mg) was obtained as a white foam; protons N, m, r, (C D 30D) is 67.29-7.08 (m, aromatic proton), 6.15 ( S, H-3), 5.18 (dd, J-2Hz, 6-H), 5.08 (d, J-5 Hz, -Cdan0Ac), 5.03 and 5.01 (2s.

-CH), 4.22 (S, NCC20), 4.17 (d, J-2Hz, H-7) , 3.81 (s.

Co　CH), 2.09 (S, -CIIOCOCdana), 0.88 (d, J- 6 Hz, CHC Dan 3) The title compound (30 mg) of Example 10 was (1.5 ml). Add anhydrous lithium iodide (133 mg) and mix The solution was stirred at 40°C under nitrogen for 48 hours. Cool the reaction mixture, filter, and trifle the residue. Washed with toluene (20ml). The filtrates were combined, evaporated and the residue was dissolved in toluene (2 0 ml) and evaporated again. This was repeated three more times. pre-product Parativ HPLC (Spherisorb 0DS-2 (25X 2 Cm) Column, eluent 38% (95:5 acetonitrile:water + trifluoroacetic acid 0.1 ml/L) and 62% (water + trifluoroacetic acid 0°1 ml/L), flow rate 15 ml/gin). Combine appropriate fractions and evaporate acetonitrile. I set it. The aqueous residue was lyophilized to give the title compound (10a+g); proton N- m, r-(CD30D) is 67.27-7.08 (1, aromatic proton).

8.20 (br s, H-3), 4.19 (s, N-C tana), 4.14 (b r　s, H-7), 2.12 (s, -CHoCOCR), 0.84 (d, J-6 Hz, CHC temperature). The mass vector [H]- is 575.2013 Measured; C28H3□N40□1 Calculated value 575.19893 6α, 7β)) 1-(4-acetyloxy-5-methyl 1]octane-4,5- Dicarboxylic acid, 4,5-dimethyl ester Compound 4B (453 mg) in dimethylformamide (51) was added to N-methylhydrogen. Droxylamine hydrochloride (156 mg) and triethylamine (0,528 m 1) and stirred for 16 hours. Dimethylformamide was evaporated under reduced pressure and the residue was partitioned between ethyl acetate (150ml) and water (100ml). The organic layer was diluted with water (10 0 ml) and brine (10011), then dried (Na2S04), Evaporated. The residue was coated with silica gel (Merck Kleselgel 60.15 0g, 240-400 mesh) and cyclohexane. : Elution was carried out with ethyl acetate 3:1 to 1:4. Combine appropriate fractions, evaporate and white The title compound (184B) was obtained as a colored foam; protons N, +s.

r, (CDCIa) is 67.36-7.08 (+a, aromatic proton), 6.00 (s, 3-H), 5.17 (d, J-2Hz, H-8 ), 5.07 (d, J-5Hz, -CIIOAc), 5D0 2 and 4.98 (2s, C-CH), 4.32 (s, NCff13), 4.1 7 (d, J-2Hz, H-7), 3.8B and 3.79 (2s, -co2Cdan) , ), 2.72 (dd, J-15 and 9Hz, LH, -CHPh), 2.1 2<s, -CHoCOCR,), 0.84 (d, J=7Hz, CHCHa) shows.

Example 13 (Is-[1α (4R, 55 lines), 3α, 4β, 5α.

* 6α, 7β]) 1-(4-hydroxy-5-methyl-3(1s-(1α(4R, 5s book), 3α, 4β, 5α.

* monotrihydroxy-3-(2-methyl-2H-tetra/-5-yl)- 2,B-dioxabicyclo[3,2゜1]octane-4,5-dicarboxylic acid (B ) ■ ff, 71) 12'' soil 2 nie 2 upper eth 2 ni ni l 3-methylene-6- phenylhexyl)-4,6,7-dolihydroxydi-3-(2-methyl-2H -tetranyl-5-yl)-2,8-dioxabicyclo[3,2゜1)mu Mutsu-Maru-Σ2 ZL Domaru Z Mouse = Sat2ヱ±A King l Chill (C) The title compound of Example 12 (184ig) was dissolved in 2.4.6-collidine (71). . Anhydrous lithium iodide (407 mg) was added and the mixture was heated at 40°C under nitrogen for 42 hours. Stirred.

The reaction mixture was filtered and the residue was washed with toluene (50owl). Filtrate and wash The liquids were combined and evaporated to dryness. Repeat this 6 times to get a residue and use it as a block. Re/Kuratave HP L C (Spherisorb 0DS-2 (25X 2 cm) column, flow rate 15ml/win, 38% (95:5 acetonitrile: water + trifluoroacetic acid 1 + al/L) and 62% (7) C 1 to 1 nofluoro vinegar Purified by acid 0.11/L)). Combine appropriate fractions and evaporate acetonitrile. I let it emanate. Lyophilization of the aqueous layer yielded three products, among which the fast tX run The reaction mixture was the title compound A (10 mg); proton nitrogen, (CD300 ) is 67.29-7.06 (m, aromatic proton).

6.02(s,)! -3), 5.38 (br　s, H-6), 4.32 (s, - NCtana), 4.15 (one of br-CHPh), 0.80 (d, J-7H z, CH (Hjs) is shown. Exact mass measured at 557.189392 ;C24H3oN40, oNa calculated value 557.185963 The title compound B (68 mg) was obtained as a white solid from the intermediate running fraction; Roton N, m, r, (CD30D) is 67.29-7.09 (m, aromatic protein roton), 6.00 (s, 3-H), 5.42 (d, J-2Hz, 6-) 1), 5.10 (d, J=5Hz, C and 0Ac), 5.01 and 4.95 (2s, C -CH2), 4.30 (s, -NCjia), 4.12 (d, J-2) 12 ．． )l-7)-2,71(dd, J-7 and 14Hz,l)1. CHPh), 2.09 (s, CHoCOCR3), 0.83 (d, J-11Hz, CHC Dan 3) is shown.

Analysis actual value: C44,93; H4,59; N7.43; H205, OCH N　O,1,5CF　Co　H・2H20 calculated value = C44,45; H4,82; N 7.15; H204, 59% off-white solid from slowest running fraction The title compound C (93 mg) was obtained as a product; proton N, ta, r, (CD3 00), 67.29-7.08 (+a, aromatic proton), 5.95 (s, 3 -t(), 5.10(br　s, -C)10Ac and H-6), 5.02 and and 4.96 (2s, C-C Dan 2).

4.32 (s, NCR), 4.12 (d, J-2Hz, H-7), 3.78 (s , -CO2C stop 3).

2.72 (dd, J-14 and 6Hz, IH, CH2Ph), 2.08 (s, -CHOCOC 3), 0.84 (d, J-6Hz, CHC 3). minutes Child ion MH+ was measured at 591, 232422; c HN O+ calculated value 591.230233 Example 14 (Is-C1α (4R, 55 pieces), 3α, 4β, 5α.

Ne The title compound (150B) of Example 2 was dissolved in anhydrous N,N-dimethylformamide (I11 1) and triethylamine (1271 g). N-methylhydroxy Ruamine hydrochloride (53 mg) was added and the resulting solution was stirred at room temperature for 16 hours.

The solvent was evaporated under vacuum and the residue was purified using preparative HP L CC3pheris. orb 0DS-2 (25X 2Cm) column, flow rate 15ml/wins maximum 40% (95 containing 1 ml/L trifluoroacetic acid) until the first component elutes. 5 acetonitrile:water) and 60% (containing 1+nl/L trifluoroacetic acid) (containing 65% trifluoroacetic acid 1 ml/L) over 2 min. 5:5 acetonitrile:water) and 35% (containing 11/L trifluoroacetic acid) It was purified by a gradient of up to 50% water). Combine appropriate fractions and reduce acetonitrile. Removed under pressure. The residual aqueous phase was lyophilized to give the title compound (72I) as a white solid. Ilg) was obtained; proton N, m, r, (CD30D) was 67.28-7.1 0 (+n, aromatic proton), 6.12 (s, H-3), 5.28 (d, J-2,5Hz, H-[i), 5.08 (d, J-58z, C tan 0Ac), 5.02 and 4.99 (2s, -CH2), 4.1IIi (d, J -2,5Hz-11-7), 2.70(dd, J-14Hz, [1l(z, C 1 of H2Ph), 2.09 (S, 0COC!L3), 0.85 (d, J- 7Hz, CH(CH) CH2Ph).

Analysis actual value: C49,52; H5,39; N 9.05CHN 0. 2. 5H20 calculated value: C49,42; H5,81, N 2-3 at 25℃ on 9.22% oatmeal agar After weeks of growth, the colony will be smoked in color on both the front and back sides of the colony. Gray-mouse gray (Rayner's Mycol.

gical Co1our Chart, 1970;Coa+1onvealt h　Published by Agricultural Bureau).

Morphological observations of the strains grown on oatmeal agar at 25°C were performed under a light microscope.

Although the fungus did not have a sexual reproductive stage, it did form powder sporophores and thus Coe. It belongs to the lomycetes. The fungus produces beak-shaped powder spore organs, and the hyphae are The conidia were smooth and had both septate and 1 septate conidia. Conidia are about 5~ 9X 1.5-3 μM (usually 7-9X 1.50-2.5 μM). chlamydospore Also, numerous multiseptate, multicellular, spherical structures resembling powder sporophyte progenitor cells were also observed.

There are no obvious reticulospores.

The isolate was identified as a species of the genus Phoma, and the identity was confirmed by CAB International. Confirmed by onal Mycological In5 titute.

In vitro results The ability of the compounds of the present invention to inhibit the squalene synthase enzyme is determined by R, M, Ta1 t, 8 nalyt, Biochem, 203, 310-31 [i (1992 ) as a substrate under the assay conditions described in This was demonstrated using phosphoric acid. Inhibition of squalene synthase is C2-”C”J farne Rat liver homogenate was combined with various concentrations of test compound in the presence of cilnylic acid. quantified by incubation.

Concentration of compound showing 50% inhibition of squalene production (14c) in 30 minute assay was taken as the engineering C5o value.

In this test, the title compounds of Examples 2.7.8 and 14 had an I It had a C5o value.

Formulation example In the examples below, the term 'active ingredient' refers to a compound of the invention, such as those mentioned in the examples above. Regarding the listed compounds.

Example 1 - Tablet a) Active ingredient 5.0mg Lactose 95.0mg Microcrystalline cellulose 90.0mg Crosslinked polyvinylpyrrolidone 8, 0mg Magnesium stearate 2.0m g Compressed weight 200.0mg Active ingredients: microcrystalline cellulose, lactose and cross-linked polyvinylpyrrolidone. 00 micron sieve and blend in a suitable mixer. Magnesium stearate sieve through a 250 micron sieve and blend with the active blend. Blend properly Compress into tablets using a suitable punch.

b) Active ingredient 5. OB Lactose 165.011g Pre-gelatinized starch 20. On+g crosslinked polyvinylpyrrolidone 8. OB Magnesium stearate 2. OB Compressed weight: 200.0g The active ingredients, lactose and pre-gelatinized starch are blended together and prepared with water. Grainy. Dry the wet mass and grind. Magnesium stearate and cross-linked poly Pass the vinylpyrrolidone through a 250 micron sieve and blend with the granules. The obtained block a) Active ingredient 5.0 mg using a suitable tablet punch Pre-gelatinized starch L93.0mg Magnesium stearate 2. OB Filling weight jl 200. Omg The active ingredient and pre-gelatinized starch were passed through a 500 micron mesh sieve. Blend with magnesium stearate (passed through a 25o Mikucon sieve). Served by Sawa. Fill the blend into appropriately sized hard gelatin capsules.

b) Active ingredient 5.0-g Lactose 177, OB Polyvinylpyrrolidone 8. OB Crosslinked polyvinylpyrrolidone 8.0mg magnesium stearate 2. OB Filling weight 200. hg The active ingredient and lactose are blended together and in a solution of polyvinylpyrrolidone. Granulate. Dry the wet mass and grind. Magnesium stearate and crosslinking The ribinylpyrrolidone is passed through a 250 micron sieve and blended with the granules. obtained Fill the blend into appropriately sized hard gelatin capsules.

Example 3 - Syrup a) Active ingredient 5.0a+g Hydroxypropyl methylcellulose 45. hg propyl hydroxybenzoate 1. 5mg butyl hydroxybenzoate 0.75mg saccharin sodium 5.0a+g Sorbitol solution 1,01 Appropriate buffer qs Appropriate flavor qs Purified water total amount 10. Until Oal Hydroxypropyl methylcellulose with hydroxybenzoate in thermally purified water and cool the solution to room temperature. Sodium saccharin, flavor and sorbitol solution to the bulk solution. Dissolve the active ingredient in some of the residual water and add to the bulk solution. Appropriate buffers control pH in the region of maximum stability. You can add it to make a difference. Adjust the solution to the desired volume, filter, and fill into a suitable container. Ru.

Active ingredient 0.1 Dextrose (anhydrous) 5.0 Benzalkonium chloride 0.02 Appropriate buffer qs Purified water up to 100% Dissolve the active ingredient and dextrose in a portion of the bulk solution. A suitable buffer is May be added to control pH in the region of maximum stability. Adjust the solution to the specified volume. Strain, filter and fill into suitable containers.

On the other hand, solutions may be provided as sterile unit doses such that preservatives are omitted from the formulation. good.

b) Sterile solution without preservatives %w/v Active ingredient 0.1 □, -1 ρCT/EP 93100487 Front page continuation (51) Int, C1,' Identification symbol Internal office reference number A61K 31/34 AED 31/70 ADZ 9454-4C C07H91028615-4C 15/10 8615-4C (81) Designation diagram EP (AT, BE, CH, DE.

DK, ES, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE) , 0A (BF, BJ, CF, CG, CI, CM, GA, GN, ML, MR, SN, TD.

TG), AT, AU, BB, BG, BR, CA, CH.

CZ, DE, DK, ES, FI, GB, HU, JP, KP, KR, KZ, L K, LU, MG, MN, MW, NL, No, NZ, PL, PT, RO , RU, SD, SE.

SK, UA, US FI (72) Inventor: Kirk, Barry, Nidward, Middlesepras, UK Benford, Berkeley, Avenue (street address) ), GRAKTS, GROUP, RESEARCH, LIMITED. (72) Inventor: Watson, Nizel Stephen Middlesepras, UK Greenford, Berkeley, Avenue (Street address) ), GRAKTS, GROUP, RESEARCH, LIMITED.

Claims (1)

[Claims] 1. Compound having the following formula (1): ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I) [In the above formula, R1 is a hydroxyl group, or [There is an array] or [There is an array] or [There is an array] or represents a group; R2 represents a hydroxyl group; R3 is ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ (In the above formula, R7 is a hydrogen atom or acetyl), [There is a sequence] (where R8 is hydrogen or hydroxyl group), [there is a sequence] and Beauty ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ represents a group selected from; R4 and R5 each independently represent a hydrogen atom or a methyl group; R6 is attached to the rest of the molecule via a ring carbon atom and optionally one of the ring nitrogen atoms. [represents a tetrazole ring substituted with a C1-4 alkyl group]2. R 2. A compound according to claim 1, wherein 4 and R5 represent a hydrogen atom. 3. R1 is the following group: ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ 3. A compound according to claim 1 or 2. . 4. Any of claims 1 to 3, wherein R6 represents an unsubstituted or methyl substituted tetrazole ring. The compound described in any one of the above. 5. R3 is the following group: ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ (wherein R7 is a hydrogen atom or an acetyl group) The compound described in any one of the above. 6. [1S-[1α (4R*, 5S*), 3α, 4β, 5α, 6α (2E, 4R *,6R*),7β]]1-(4-acetyloxy-5-methyl-3-methylene -6-phenylhexyl)-4,6,7-trihydroxy-3-(1H-tetra sol-5-yl)-2,8-dioxabicyclo[3.2.1]octane-4, 5-dicarboxylic acid, 6-(4,6-dimethyl-2-octenoate); [1S-[1α (4R*, 5S*), 3α, 4β, 5α, 6α (2E, 4R*, 6R*),7β]]1-(4-acetyloxy-5-methyl-3-methylene-6 -phenylhexyl)-4,6,7-trihydroxy-3-(2-methyl-2H -tetrazol-5-yl)-2,8-dioxabicyclo[3.2.1]okuku -4,5-dicarboxylic acid, 6-(4,6-dimethyl-2-octenoate); [1S-[1α (4R*, 5S*), 3α, 4β, 5α, 6α, 7β]] 1-( 4-acetyloxy-5-methyl-3-methylene-6-phenylhexyl)-4 ,6,7-trihydroxy-3-(1H-tetrazol-5-yl)-2,8- dioxabicyclo[3.2.1]octane-4,5-dicarboxylic acid; and [1S-[1α (4R*, 5S*), 3α, 4β, 5α, 6α, 7β]] 1-( 4-acetyloxy-5-methyl-3-methylene-6-phenylhexyl)-4 ,6,7-trihydroxy-3-(2-methyl-2H-tetrazol-5-yl )-2,8-dioxabicyclo[3.2.1]octane-4,5-dicarboxylic acid and their salts and protected derivatives. 7. 7. A compound according to any one of claims 1 to 6 for use in therapy. 8. Diseases in which lowering plasma cholesterol levels is beneficial in animals, including humans 7. A compound according to any one of claims 1 to 6 for use in the treatment of. 9. Claim 1: Used for the treatment of fungal infections in human or non-human animal patients. 7. The compound according to any one of . 10. Diseases associated with hypercholesterolemia and/or hyperlipoproteinemia Treatment of disease or true OCOC H-ECHCH(CH3)(CH2)3CH3, -OCOCH-ECHC(CH 3) -ECHCH(CH3)CH2CH3 or -OCO-X-CH2CH(C H3) CH2CH3 (where ・X is -CH-ECHCH(CH3)-, -CH2 CH(OH)CH(CH3)-, -CH-ECHC(OH)(CH3)-, -C Treat bacterial diseases with H2CH(OH)CH2- or -CH2CH2CH(CH3)- A method for treating a human or non-human animal individual, the method comprising: A compound according to any one of claims 1 to 6 which inhibits squalene synthase. A method comprising administering an effective amount to said individual. 11. Any one of claims 1 to 6 together with one or more carriers and/or excipients. A pharmaceutical composition comprising a compound according to item 1. 12. Reducing plasma cholesterol levels in animals, including humans, is beneficial An active amount of a compound according to any one of claims 1 to 6 used in the treatment of a condition. A pharmaceutical composition comprising. 13. The claim is for use in the treatment of fungal infections in human or non-human animal patients. 7. A pharmaceutical composition comprising an active amount of a compound according to any one of 1 to 6. 14. Suitable for oral, buccal, topical, parenteral, implantable, rectal, ocular or genitourinary administration. claim 11, in a form suitable for administration by inhalation or insufflation. 14. The pharmaceutical composition according to any one of items 1 to 13. 15. 15. A pharmaceutical composition according to any one of claims 11 to 14 in unit dosage form. 16. Hypercholesterolemia and/or in human or non-human animal patients Any one of claims 1 to 6 for the manufacture of a medicament for treating hyperriboproteinemia. USE OF THE COMPOUNDS DESCRIBED IN SECTION. 17. For the manufacture of medicaments for the treatment of fungal infections in human or non-human animal patients. , use of a compound according to any one of claims 1 to 6. 18. A method for producing the compound according to claim 1, comprising: (A) (for the production of compounds of formula (I) in which R6 represents an unsubstituted tetrazole ring) Compounds of the following formula (II): ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (II) (the above formula (wherein, R1 to R3 have the same meanings as in claim 1, and R4a and R5a are protecting groups) is reacted with an azide source at elevated temperature, followed by removal of any protecting groups present if necessary. ;or (B) a compound of formula (I) or a protected derivative thereof with a different compound of formula (I) or converting it into a protected derivative thereof and subsequently removing any protecting groups present if necessary; A method consisting of. 19. A compound of formula (II). 20. A compound of formula (V). 21. A compound according to any one of claims 1 to 6 substantially as herein described. thing. 22. 16. A method according to any one of claims 11 to 15 substantially as herein described. Composition.