WO2010131691A1 - ポリケチド化合物 - Google Patents
ポリケチド化合物 Download PDFInfo
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- WO2010131691A1 WO2010131691A1 PCT/JP2010/058076 JP2010058076W WO2010131691A1 WO 2010131691 A1 WO2010131691 A1 WO 2010131691A1 JP 2010058076 W JP2010058076 W JP 2010058076W WO 2010131691 A1 WO2010131691 A1 WO 2010131691A1
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- compound
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- ringworm
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- capnodiaceae
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- DOMXUEMWDBAQBQ-WEVVVXLNSA-N CC(C)(C)C#C/C=C/CN(C)Cc1cccc2ccccc12 Chemical compound CC(C)(C)C#C/C=C/CN(C)Cc1cccc2ccccc12 DOMXUEMWDBAQBQ-WEVVVXLNSA-N 0.000 description 1
- PCXGABLZLXTHMH-UHFFFAOYSA-N CCC(C)CC(C)CCC(CCC1)OC1C(C(N(C)C=C1C(C(C(C2O)O)O)C2O)=O)=C1O Chemical compound CCC(C)CC(C)CCC(CCC1)OC1C(C(N(C)C=C1C(C(C(C2O)O)O)C2O)=O)=C1O PCXGABLZLXTHMH-UHFFFAOYSA-N 0.000 description 1
- IBHQZHNSPLOUKU-UHFFFAOYSA-N CCC(C)CC(C)CCC(CCC1)OC1C(C(N(C)C=C1C(CC(C2O)O)C2O)=O)=C1O Chemical compound CCC(C)CC(C)CCC(CCC1)OC1C(C(N(C)C=C1C(CC(C2O)O)C2O)=O)=C1O IBHQZHNSPLOUKU-UHFFFAOYSA-N 0.000 description 1
- VHVPQPYKVGDNFY-UHFFFAOYSA-N CCC(C)N(C1=O)N=CN1c(cc1)ccc1N(CC1)CCN1c(cc1)ccc1OCC1OC(C[n]2ncnc2)(c(c(Cl)c2)ccc2Cl)OC1 Chemical compound CCC(C)N(C1=O)N=CN1c(cc1)ccc1N(CC1)CCN1c(cc1)ccc1OCC1OC(C[n]2ncnc2)(c(c(Cl)c2)ccc2Cl)OC1 VHVPQPYKVGDNFY-UHFFFAOYSA-N 0.000 description 1
- DDUHZTYCFQRHIY-RBHXEPJQSA-N C[C@H](CC(C=C1OC)=O)[C@]1(C1=O)Oc2c1c(OC)cc(OC)c2Cl Chemical compound C[C@H](CC(C=C1OC)=O)[C@]1(C1=O)Oc2c1c(OC)cc(OC)c2Cl DDUHZTYCFQRHIY-RBHXEPJQSA-N 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
- C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
- C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
- A61K31/4433—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a six-membered ring with oxygen as a ring hetero atom
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/10—Antimycotics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P33/00—Antiparasitic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P33/00—Antiparasitic agents
- A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P33/00—Antiparasitic agents
- A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
- A61P33/08—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis for Pneumocystis carinii
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/16—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing two or more hetero rings
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
Definitions
- the present invention relates to a pharmaceutical composition, particularly a cyclic compound useful as an active ingredient of a pharmaceutical composition for treating ringworm, protozoal infection, and / or Pneumocystis pneumonia.
- onychomycosis is caused by infecting the nails with ringworm fungus (aquatic fungus). Since tinea pedis, a so-called athlete's foot, has been left untreated, it often develops due to the migration of ringworm bacteria from the skin of the foot into the nail, and the main symptoms are clouding and thickening of the nail.
- ringworm fungus aquatic fungus
- tinea pedis a so-called athlete's foot
- Terbinafine has been reported to have weak activity against dormant bacteria present in the vicinity of the infection (Mycoses, 46, pp.506-510, 2003). Long-term administration of drugs is essential to treat onychomycosis. In the nail of onychomycosis, bacteria that are growing, that is, mycelial elongation, and bacteria that are dormant, that is, spores and mycelial growth are stopped are mixed. In the nail penetrated by Terbinafine, the growing fungus is killed, so the ringworm does not infect the newly created nail. Since dormant bacteria are not sterilized by Terbinafine, the cure of onychomycosis requires administration for about 6 months until the nail changes. Therefore, if a dormant bacterium can be sterilized, it can be sterilized before the nail grows, so that it can be expected to shorten the treatment period and show a high cure rate.
- the compound having a 4-hydroxy-2-one skeleton for example, Funiculosin (formula D, however, wherein --- is a double bond), Tetrahydrofuniculosin (formula D, In the formula, - - is a single bond), there is Ilicicolin H (formula E) or the like.
- Funiculosin is an antifungal active substance isolated from a culture solution of Penicillium funiculosum (Non-Patent Documents 1, 2, etc.), and Tetrahydrofuniculosin is a compound semi-synthesized from Funiculosin (Patent Document 1, Non-Patent Documents 3 and 4).
- Ilicicolin H is an antifungal active substance isolated from the culture solution of the incomplete bacterium Cylindrocladium iliciola MFC-870 (Non-patent Documents 5 and 6).
- a compound useful as an active ingredient of a pharmaceutical composition a pharmaceutical composition for treating ringworm, protozoal infection, and / or Pneumocystis pneumonia.
- Capnodiaceae strain No. 339855 Capnodiaceae strain ⁇ ⁇ No. 339855
- the present inventors have succeeded in isolating this polyketide compound having an excellent antibacterial action from the culture solution of this strain, and completed the present invention.
- the present invention provides a pharmaceutical composition comprising a compound of formula (I) or a salt thereof, and a compound of formula (I) or a salt thereof, and an excipient, in particular, ringworm, protozoal infection, and
- the present invention relates to a pharmaceutical composition that is a preventive or therapeutic agent for Pneumocystis pneumonia.
- the invention provides a compound of formula (I) or a salt thereof for use in the prevention or treatment of ringworm, protozoal infection, and / or Pneumocystis pneumonia, ) Or a salt thereof, and a method for the treatment of ringworm, protozoan infection, and Pneumocystis pneumonia.
- the present invention also includes culturing Capnodiaceae microorganisms having the ability to produce the compound of formula (I), preferably Capnodiaceae strain No. 339855 (Capnodiaceae strain No. 339855) or mutants thereof. And a method for producing a compound of (I) or a salt thereof, comprising isolating the compound of formula (I) or a salt thereof from the culture.
- the present invention relates to a capnody deposited as NITE BP-735, a deposit number, NITE BP-735, which is a novel microorganism having the ability to produce the compound of formula (I), which is the National Institute for Product Evaluation Technology Patent Microorganism Depositary. It also relates to Aceae strain No.339855 (Capnodiaceae strain No.339855) or its mutants.
- the compound of formula (I) or a salt thereof may be used to treat ringworm, such as ringworm granuloma, onychomycosis, hand / foot ringworm, body ringworm, hip ringworm, hairy coat ringworm, head Ringworm, Celsus pressure ulcer, tinea dermatosis, acute hairy deep tinea, and / or acute hairy deep tinea, etc.), protozoal infections (eg, malaria, toxoplasmosis), And / or can be used as a preventive and / or therapeutic agent for Pneumocystis pneumonia (formerly Carini pneumonia).
- ringworm such as ringworm granuloma, onychomycosis, hand / foot ringworm, body ringworm, hip ringworm, hairy coat ringworm, head Ringworm, Celsus pressure ulcer, tinea dermatosis, acute hairy deep tinea, and / or acute hairy deep tinea, etc.
- protozoal infections
- FIG. 1 is a diagram showing the 1 H-NMR spectrum (measurement solvent: CDCl 3 ) of Compound A.
- FIG. 2 is a diagram showing a 13 C-NMR spectrum (measurement solvent: CDCl 3 ) of Compound A.
- FIG. 3 is a diagram showing the 1 H-NMR spectrum (measurement solvent: CDCl 3 ) of Compound B.
- FIG. 4 is a diagram showing a 13 C-NMR spectrum of Compound B (measuring solvent: CDCl 3 ).
- the compound of the present invention may form a salt with a base.
- Specific examples of such salts include inorganic bases such as sodium, potassium, magnesium and calcium.
- the salt in the present invention includes so-called complex salts and chelate compounds.
- the compound of the formula (I) has 7 or 8 asymmetric carbon atoms, and optical isomers based on this can exist.
- the present invention encompasses both compounds of formula (I) isolated as a single optical isomer or compounds of formula (I) obtained as a mixture of isomers.
- the present invention includes pharmaceutically acceptable prodrugs of the compound of formula (I).
- Pharmaceutically acceptable prodrugs are for example those skilled in the art that one or more —OH possessed by a compound of formula (I) can be converted to the original —OH by solvolysis or under physiological conditions. Prodrugs that are chemically modified with well-known groups.
- the present invention also includes various hydrates and solvates of the compound of formula (I) and salts thereof, and crystalline polymorphic substances.
- the present invention also includes compounds labeled with various radioactive or non-radioactive isotopes.
- Capnodiaceae strain No.339855 Capnodiaceae strain No.339855
- a type of fungus was isolated from the fallen leaves of zelkova collected in Yoro Valley, Otaki-cho, Isumi-gun, Chiba Prefecture, by the wet chamber method. It was done.
- This strain has been deposited in Japan with the National Institute for Product Evaluation Technology Patent Microorganisms Deposit Center (2-5-8 Kazusa-Kamashita, Kisarazu City, Chiba Prefecture, 292-0818, Japan) with the accession number NITE P-735 (date of access: 2009) April 16).
- NITE P-735 date of access: 2009
- the growth temperature of this bacterium was 12 ° C to 30 ° C, and the optimal growth temperature was 20 ° C to 25 ° C. These growth temperature data were measured on potato dextrose agar (Difco 2010). The growth possible pH was 2.02 to 11.46, and the optimum growth pH was 5.45 to 6.15. The viable pH was measured with YM broth (Difco).
- Morphological features were determined based on culture on corn meal agar.
- the vegetative mycelium was brown and the thickness was 2.3 to 4.0 ⁇ m. No thick film spores were observed.
- the conidial pattern was not branched and formed a determinate synnemata.
- the total length was 200 to 260 mm, and the thickness was 8 to 10 mm.
- the base of the synnema spread to form a stroma, and the width was 30-40 ⁇ m.
- the morphology is similar to the genus Graphium, but the mycelial bundle was slightly swollen at the top of the synnema and narrowed again at the tip. Furthermore, several setae (about 20 ⁇ m) were formed so as to surround the tip.
- Colorless conidia were formed in the narrowed tip, and dropped into the upper part of the cinema.
- the conidia were elliptical to cylindrical and had a size of (3.2 to 3.6) ⁇ (1.7 to 1.76) ⁇ m.
- the surface was smooth with an optical microscope (400 ⁇ ).
- This bacterium formed a cinema and the tip (upper part) swelled. Furthermore, several short setas characteristic of this bacterium were formed at the swollen tip. Since this bacterium is morphologically similar to the genus Graphium, homology search was performed using 28S rDNA. As a result, the present bacterium was close to the genus Capnodium, and the result was that it was different from the multiple strains of the genus Graphium. At present, this bacterium has been identified as one strain of Capnodiaceae (Capnodiaceae) and named as Capnodiaceae strain (No.339855).
- Potato dextrose agar medium Sabouraud dextrose agar medium, oatmeal agar medium, malt extract agar medium, Czapek's agar medium, and MY20 agar medium showed almost similar properties. Therefore, the characteristics of potato dextrose agar medium as described above (2 ). In addition, since spore formation was confirmed only on the corn meal agar medium, the culture characteristics on the corn meal agar medium were also described in (2) above as a characteristic medium.
- Capnodiaceae strain No. 339855 Capnodiaceae strain No. 339855 used in the present invention is not only a microorganism isolated from nature, This includes those artificially mutated with ultraviolet rays, radiation, chemical agents, etc., and natural mutants thereof.
- the compound of the present invention is obtained by culturing a microorganism of the family Capnodiaceae having the ability to produce the compound of the present invention, preferably Capnodiaceae strain No. 339855 (Capnodiaceae strain No. 339855) or a mutant thereof. It is obtained by isolating the compound of the present invention from The culture is performed according to a general microorganism culture method.
- the medium may be any medium containing a nutrient source used by Capnodiaceae strain No. 339855 (capnodiaceae strain No. 339855), and a synthetic medium, semi-synthetic medium or natural medium is used.
- the composition of the medium is, for example, L-arabinose, D-xylose, D-glucose, D-fructose, sucrose, inositol, L-rhamnose, raffinose, D-mannitol, mannose, melibiose, lactose, D- Galactose, maltose, trehalose, salicin, xanthine, chitin, starch, glucose, dextrin, glycerin, vegetable oil, etc., as nitrogen sources are meat extract, peptone, gluten meal, cottonseed meal, soy flour, peanut flour, fish meal, corn steep Liquor, dry yeast, yeast extract, ammonium chloride, ammonium sulfate,
- metal salts such as sodium, potassium, magnesium, calcium, zinc and cobalt, nitrates, carbonates, phosphates and the like are added as necessary. Furthermore, if necessary, a production promoting substance or an antifoaming agent such as methionine, cysteine, cystine, thiosulfate, methyl oleate, lard oil, silicone oil, and surfactant can be added.
- the culture conditions it is generally advantageous to culture under aerobic conditions, and the culture temperature is in the range of 12 ° C to 30 ° C, preferably in the vicinity of 20 ° C to 25 ° C.
- the culture period is appropriately set according to the composition of the medium and temperature conditions, but is usually about 1 to 30 days, preferably about 2 to 7 days.
- a method of purifying and isolating a physiologically active substance from a normal microorganism culture is applied. That is, the culture is extracted with a suitable organic solvent, and the active substance is isolated from the extract by purification. That is, it is separated and purified by means used for the production of a general physiologically active substance using the difference in solubility and solubility in an appropriate solvent using antifungal activity as an index. These methods can be used alone as needed, or can be combined and repeated in any order.
- Other purification methods include removing the cells by culturing the culture as it is, or centrifuging or filtering, and then the difference in solubility and solubility in an appropriate solvent, the difference in the precipitation rate from the solution, and various adsorbents.
- a method using a difference in adsorption affinity, a difference in distribution between two liquid phases, and the like can be applied. Specifically, for example, a method of purifying the compound by bringing the culture solution into contact with an appropriate carrier to adsorb the compound and then eluting with an appropriate solvent can be mentioned. These methods can be used alone as needed, or can be combined and repeated in any order.
- the compound of the formula (I) or a salt thereof of the present invention can be obtained by a conventional method from a culture in which the substance-producing microorganism is cultured in a nutrient medium and the compound is accumulated.
- Any microorganism can be used as long as it is a microorganism belonging to the Capnodiaceae family and capable of producing the compound.
- Capnodiaceae strain No. 339855 (Capnodiaceae strain No. 339855) deposited internationally as NITE BP-735 can be used.
- a pharmaceutical composition containing one or more of the compounds of formula (I) or a salt thereof as an active ingredient is an excipient usually used in the art, that is, a pharmaceutical excipient or a pharmaceutical carrier. Can be prepared by a commonly used method.
- Administration is orally by tablets, pills, capsules, granules, powders, liquids, etc., or injections such as intra-articular, intravenous, intramuscular, suppositories, eye drops, ophthalmic ointments, transdermal solutions, Any form of parenteral administration such as an ointment, a transdermal patch, a transmucosal liquid, a transmucosal patch, and an inhalant may be used.
- Tablets, powders, granules, etc. are used as solid compositions for oral administration.
- one or more active ingredients are combined with at least one inert excipient such as lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, starch, polyvinylpyrrolidone. And / or mixed with magnesium aluminate metasilicate.
- the composition may contain an inert additive, for example, a lubricant such as magnesium stearate, a disintegrant such as sodium carboxymethyl starch, a stabilizer, or a solubilizing agent according to a conventional method. .
- tablets or pills may be coated with a sugar coating or a film of a gastric or enteric substance.
- Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or elixirs and the like, and commonly used inert diluents such as purified water. Or contains ethanol.
- the liquid composition may contain solubilizers, wetting agents, auxiliaries such as suspending agents, sweeteners, flavors, fragrances and preservatives in addition to the inert diluent.
- the injection for parenteral administration contains a sterile aqueous or non-aqueous solution, suspension or emulsion.
- aqueous solvent include distilled water for injection or physiological saline.
- non-aqueous solvents include propylene glycol, polyethylene glycol or vegetable oils such as olive oil, alcohols such as ethanol, or polysorbate 80 (a pharmacopeia name).
- Such compositions may further contain isotonic agents, preservatives, wetting agents, emulsifying agents, dispersing agents, stabilizing agents, or solubilizing agents. These are sterilized by, for example, filtration through a bacteria-retaining filter, blending with a bactericide or irradiation. These can also be used by producing a sterile solid composition and dissolving or suspending it in sterile water or a sterile solvent for injection before use.
- External preparations include ointments, plasters, creams, jellies, poultices, sprays, lotions, eye drops, eye ointments and the like.
- ointment bases include commonly used ointment bases, lotion bases, aqueous or non-aqueous solutions, suspensions, emulsions, and the like.
- ointment or lotion bases include polyethylene glycol, propylene glycol, white petrolatum, white beeswax, polyoxyethylene hydrogenated castor oil, glyceryl monostearate, stearyl alcohol, cetyl alcohol, lauromacrogol, sorbitan sesquioleate, etc. Can be mentioned.
- a transmucosal agent such as an inhalant or a nasal agent
- a solid, liquid or semi-solid agent is used and can be produced according to a conventionally known method.
- known excipients and further pH adjusters, preservatives, surfactants, lubricants, stabilizers, thickeners and the like may be added as appropriate.
- an appropriate device for inhalation or insufflation can be used.
- a known device such as a metered dose inhalation device or a nebulizer
- the compound is administered alone or as a powder in a formulated mixture or as a solution or suspension in combination with a pharmaceutically acceptable carrier. be able to.
- the dry powder inhaler or the like may be for single or multiple administrations, and a dry powder or a powder-containing capsule can be used. Alternatively, it may be in the form of a pressurized aerosol spray using a suitable propellant, for example, a suitable gas such as chlorofluoroalkane, hydrofluoroalkane or carbon dioxide.
- a suitable propellant for example, a suitable gas such as chlorofluoroalkane, hydrofluoroalkane or carbon dioxide.
- the daily dose is about 0.01 to 100 mg / kg per body weight, preferably 0.1 to 10 mg / kg. Administer in divided doses.
- the daily dose is suitably about 0.01 to 100 mg / kg per body weight, and is administered once to several times a day. The dose is appropriately determined according to individual cases in consideration of symptoms, age, sex, and the like.
- the compound of the formula (I) can be used in combination with various therapeutic agents or preventive agents for diseases for which the compound of the formula (I) is considered to be effective.
- the combination may be administered simultaneously, separately separately, or at desired time intervals.
- the simultaneous administration preparation may be a compounding agent or may be separately formulated.
- the manufacturing method of the compound of Formula (I) is demonstrated in detail.
- this invention is not limited to the compound as described in the following Example.
- the production method of the compound of the formula (I) is not limited to the production methods of the specific examples shown below, and the compound of the formula (I) may be a combination of these production methods or a person skilled in the art. It can also be produced by methods that are self-evident.
- MeCN acetonitrile
- MeOH methanol
- MgSO 4 .7H 2 O magnesium sulfate heptahydrate
- t-BuOH tert-butyl alcohol
- TFA trifluoroacetic acid
- HR ESI MS high resolution electrospray mass spectrometry
- Example 1 (Culture production of Compound A and Compound B) Dispense seed medium (30 mL; 2% corn starch, 1% glycerin, 1% sucrose, 1% pharmame media, 1% gluten meal, 0.2% Tween 80) into Erlenmeyer flask (size: 100 mL) And sterilized in an autoclave (121 ° C., 30 minutes). Slope culture (slant culture) of Capnodiaceae strain No.339855 (Caprantiace strain No.339855) is inoculated aseptically into this seed medium and aseptically inoculated on a rotary shaker at 25 ° C for 4 days. Shake culture (220 rpm).
- production medium ⁇ 20 mL; 4% glucose, 6% soluble starch (manufactured by Nacalai Tesque), 2% yeast extract (manufactured by Wako Pure Chemical Industries, Ltd.), 0.02% KCl, 0.02% MgSO 4. 7H 2 O, 1% K 2 HPO 4 , 1% ⁇ -cyclodextrin and 2% agar) were sterilized by an autoclave and dispensed into a petri dish (diameter 9 cm). A seed culture (2 mL) was aseptically applied to the petri dish and cultured at 25 ° C. for 10 days.
- Example 2 The antifungal activity of the compound of the present invention was confirmed by the following method.
- the antifungal activity against the following test bacteria was measured using a micro liquid dilution method (by Kume Mitsuko, Toshikazu Yamazaki, Clinical and Microbiology, Vol. 21, No. 5, pp. 573-580, 1994).
- Compound A exhibited the activity shown below.
- Table 1 shows the minimum inhibitory concentration (MIC) of Compound A.
- ringworm specifically, for example, ringworm granuloma, onychomycosis, tinea pedis, body ringworm, hip ringworm, hairy hair ringworm, head ringworm, Celsus acne It has been shown to be useful for the treatment of trichodermatosis, acute hairy deep tinea, and / or acute hairy deep tinea.
- Example 3 (Complex III inhibitory activity assay) Medium (0.1% glucose, 0.2% bactotryptone, 0.2% K 2 HPO 4 , 0.005% MgSO 4 .7H 2 O, 0.005% CaCl 2 .2H 2 O) in an Erlenmeyer flask The aliquot was sterilized by autoclaving. Trichophyton mentagrophytes FP2103 conidia were inoculated into this medium and cultured with shaking on a rotary shaker at 30 ° C. for 40 hours.
- the bacterial cells are collected from this culture solution, and the bacterial cells are lysed with cell wall lysing enzyme solution ⁇ 1M KCl, 25 mM CaCl 2 ⁇ 2H 2 O, 10 mM MgCl 2 ⁇ 6H 2 O, 5 mg / mL zymolyce 100T ) 1 mg / mL chitinase (manufactured by Sigma), 5 mg / mL driselase (manufactured by Sigma) ⁇ and shaken at 30 ° C. for 4 hours.
- the enzyme-treated solution was filtered through a funnel packed with gauze to obtain a protoplast.
- Mitochondrial fraction was prepared from protoplasts using Mitochondria Isolation Kit (BioChain Institute, Inc.). Complex III inhibitory activity was measured by ubiquinol-cytochrome c reductase assay using the adjusted mitochondrial fraction (The Journal of Biological Chemistry, 279, pp. 8708-8714, 2004). As a result, Compound A inhibited complex III at 0.9 ng / mL.
- the compound of the formula (I) or a salt thereof has a complex III inhibitory action. Therefore, it was suggested that it can be used for the treatment of protozoan infections (for example, malaria, toxoplasmosis) and / or Pneumocystis pneumonia.
- protozoan infections for example, malaria, toxoplasmosis
- Pneumocystis pneumonia for example, malaria, toxoplasmosis
- Example 4 Cytotoxicity
- EL-4 cells a mouse T lymphoma cell line
- test drugs for cytotoxicity, EL-4 cells, a mouse T lymphoma cell line, were added with test drugs at various concentrations, cultured at 37 ° C for 72 hours in a CO 2 incubator, and then cell counting kit (manufactured by Wako Pure Chemical Industries, Ltd.). ), The number of cells was measured, and the IC 50 value was calculated and evaluated.
- Compound A showed an IC 50 value of 6.2 ⁇ g / mL against EL-4 cells, and showed no cytotoxicity at a concentration of 3.1 ⁇ g / mL or less.
- the compound of formula (I) or a salt thereof may be used to treat ringworm, such as ringworm granuloma, onychomycosis, hand / foot ringworm, body ringworm, hip ringworm, hairy coat ringworm, head Ringworm, quercus acne, tinea variola, acute hairy deep tinea, and / or acute hairy deep tinea, etc.), protozoal infections (eg, malaria, toxoplasma), and It can be used as a preventive and / or therapeutic agent for Pneumocystis pneumonia (formerly Carini pneumonia).
- ringworm such as ringworm granuloma, onychomycosis, hand / foot ringworm, body ringworm, hip ringworm, hairy coat ringworm, head Ringworm, quercus acne, tinea variola, acute hairy deep tinea, and / or acute hairy deep tinea, etc.
- protozoal infections eg, malaria, tox
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Abstract
Description
爪白癬症の治療薬としては、スクワレンエポキシダーゼ阻害作用を有するTerbinafine(下記式A)、微小管の機能を阻害するGriseofulvin(下記式B)、チトクロームP450阻害作用を有するItraconazole(下記式C)が用いられている。
Funiculosinは、Penicillium funiculosum の培養液から単離された抗真菌活性物質であり(非特許文献1、2等)、TetrahydrofuniculosinはFuniculosinから半合成した化合物である(特許文献1、非特許文献3,4)。また、Ilicicolin Hは、不完全菌Cylindrocladium iliciola MFC-870の培養液から単離された抗真菌活性物質である(非特許文献5,6)。
本発明化合物は、塩基との塩を形成する場合もある。かかる塩としては、具体的には、ナトリウム、カリウム、マグネシウム、カルシウム等の無機塩基等が挙げられる。また、本発明における塩は、いわゆる錯塩やキレート化合物も包含する。
(1)生産菌の由来
真菌の一種である、カプノディアセアエ ストレインNo.339855(Capnodiaceae strain No.339855)は千葉県夷隅郡大多喜町の養老渓谷で採集したケヤキの落葉から湿室法により分離された。本菌株は独立行政法人製品評価技術基盤機構特許微生物寄託センター(〒292-0818 千葉県木更津市かずさ鎌足2-5-8)に国内寄託し、受託番号NITE P-735(受託日:2009年4月16日)が付与された。さらに、2010年1月29日(移管日)にブダペスト条約に基づく国際寄託への移管請求が受領され、受託番号NITE BP-735が付与された(受託日:2009年4月16日)。
ポテトデキストロース寒天培地(Difco社製 2010)上での生育は良好で、25℃で2週間後の直径は23~26 mmであった。集落表面はフェルト状(felty)で、部分的に羊毛状(floccose)であった。集落周縁は円状(entire)であり、中央部から周縁部に数本の放射状の溝を生じた。集落の色はフェルト状の部分が灰緑色(grayish green, 30F8)で羊毛状の部分が薄灰緑色(grayish green, 29E4)であった。集落裏面の色は全体的に濃灰緑色(grayish green, 28F4)であった。30℃での2週間後の集落の直径は6~7 mmであった。5℃と37℃では生育は認められなかった。色調の記載はMethuen Handbook of Colour(Kornerup, A. and J.H. Wanscher, 3rd ed., pp.252, Methuen, London, 1987年)に従った。
培養特性は、市販の培地、および文献に記載された組成で調製した培地で判定した。ポテトデキストロース寒天培地はDifco社 2010、サブローデキストロース寒天培地はDifco社 0109、Emerson YpSs寒天培地はDifco社0739、コーンミール寒天培地はDifco社0386、オートミール寒天培地はDifco社 0552をそれぞれ購入した。麦芽抽出寒天培地、ツァペック氏液寒天培地、MY20寒天培地の組成は、JCMカタログ(Japan Collection of Microorganisms, the Institute of Physical and Chemical Research, Saitama, Nakase,T. 6th ed. ,pp.617,1995年)に従った。
本発明化合物は、本発明化合物の生産能を有するカプノディアセアエ科の微生物、好ましくは、カプノディアセアエ ストレインNo.339855(Capnodiaceae strain No.339855)もしくはその変異体を培養し、その培養物から本発明化合物を単離することによって得られる。培養は、一般微生物の培養方法に準じて行われる。
MeCN=アセトニトリル、MeOH=メタノール、MgSO4・7H2O=硫酸マグネシウム7水和物、t-BuOH=tert-ブチルアルコール、TFA=トリフルオロ酢酸、HR ESI MS=高分解能エレクトロスプレー法質量分析
(化合物Aおよび化合物Bの培養生産)
種培地(30 mL;2% コーンスターチ、1% グリセリン、1% シュークロース、1% ファーマメディア、1% グルテンミール、0.2% Tween 80を含有)を三角フラスコ(サイズ:100 mL)に分注し、オートクレーブで滅菌した(121℃、30分)。この種培地にカプノディアセアエ ストレインNo.339855(Capnodiaceae strain No.339855)の斜面培養物(スラント培養物)を一白金耳分、無菌的に植菌し、25℃で4日間、ロータリーシェーカーで振盪培養した(220 rpm)。次に、生産培地{20 mL;4% グルコース、6% 可溶性デンプン(ナカライテスク社製)、2% 酵母エキス(和光純薬工業社製)、0.02% KCl、0.02% MgSO4・7H2O、1% K2HPO4、1% β‐サイクロデキストリン、2% 寒天を含有)をオートクレーブで滅菌し、シャーレ(直径9 cm)に分注した。このシャーレに種培養物(2 mL)を無菌的に塗布し、25℃で10日間培養した。
上記の培養方法で得られた培養物(1.8 L)をアセトン(2 L)の中に浸し、1時間撹拌後、ろ過した。ろ液に等量の水を加え、Diaion HP20カラム(100×16 mm I.D.、サイズ:20 mL、三菱ケミカル社製)に通液し、混合溶媒{アセトン:水=60:40(v/v)、100 mL}で溶出した。溶出液に水(100 mL)を加え、Daisogel SP-120-ODS-Bカラム(サイズ:180 mL、250×30mm I.D.DAISO社製)に通液し、混合溶媒{MeCN:水=50:50(v/v)(0.05% TFAを含む)}を移動相として、化合物Aと化合物Bを含むフラクションを各々分取した。化合物Aを含むフラクションに等量の水を加え、OASIS HLB 固相抽出カートリッジ(サイズ:6g/35cc、Waters社製)に通液し、MeCN(100 mL)で溶出した。溶出液を減圧濃縮し、t-BuOHを添加し、凍結乾燥することで、化合物A(265 mg)を粉末として得た。
色および形状 = 白色粉末
旋光度 = [α]D 25 -120°(c 0.5,MeOH)
分子式 = C25H41NO7
HR ESI MS(m/z) = Calcd for C25H42N1O7 [M+H]+468.2961, Found 468.2965
UV = λMeOHnm (ε):233(sh), 290.5(6,100)
1H-NMRスペクトラム = 図1に示した。
13C-NMRスペクトラム = 図2に示した。
色および形状 = 白色粉末
分子式 = C25H41NO6
HR ESI MS(m/z) = Calcd for C25H42N1O6 [M+H]+452.3012,Found 452.3011
1H-NMRスペクトラム = 図3に示した。
13C-NMRスペクトラム = 図4に示した。
(抗真菌活性)
本発明化合物の抗真菌活性は以下の方法で確認した。
下記検定菌に対する抗真菌活性は、微量液体希釈法(久米光、山崎敏和著、臨床と微生物、21巻5号、pp.573-580、1994年)を用いて測定した。例えば、化合物Aは以下に示す活性を示した。表1に化合物Aの最小生育阻止濃度(MIC)を示す。
(Complex III 阻害活性測定法)
培地(0.1% グルコース、0.2% バクトトリプトン、0.2% K2HPO4、0.005% MgSO4・7H2O、0.005% CaCl2・2H2O)を三角フラスコに分注し、オートクレーブで滅菌した。この培地にTrichophyton mentagrophytes FP2103の分生子を植菌し、30℃で40時間、ロータリーシェーカーで振盪培養した。この培養液から菌体を回収し、菌体を細胞壁溶解酵素液{1M KCl、25mM CaCl2・2H2O、10mM MgCl2・6H2O、5mg/mL ザイモリエイス-100T(生化学工業株式会社製)、1mg/mL chitinase(Sigma社製)、5mg/mL driselase(Sigma社製)}に浸し、30℃で4時間振盪した。この酵素処理液を、ガーゼを詰めたロートでろ過してプロトプラストを得た。Mitochondria Isolation Kit(BioChain Institute, Inc.)を用いてプロトプラストからミトコンドリア画分を調整した。Complex III阻害活性は調整したミトコンドリア画分を用いてubiquinol-cytochrome c reductase assayにより測定した(The Journal of Biological Chemistry,279,pp.8708-8714,2004年)。その結果、化合物Aはcomplex IIIを0.9 ng/mLで阻害した。
(細胞傷害性)
細胞障害性は、マウスTリンパ腫細胞株であるEL-4細胞に試験薬剤を各種濃度で添加し、CO2インキュベーターで37℃、72時間培養した後、セルカウンティングキット(和光純薬工業株式会社製)で細胞数を測定し、IC50値を算出して評価した。その結果、例えば、化合物Aは、EL-4細胞に対して、IC50値は、6.2 μg/mLであり、3.1 μg/mL以下の濃度において細胞障害性は示さなかった。
Claims (10)
- RがOHである、請求項1に記載の化合物又はその塩。
- RがHである、請求項1に記載の化合物又はその塩。
- 請求項1に記載の化合物またはその塩、および製薬学的に許容される賦形剤を含有する医薬組成物。
- 白癬症、原虫感染症、及び/又は、ニューモシスチス肺炎の予防若しくは治療剤である請求項4記載の医薬組成物。
- 白癬症、原虫感染症、及び/又は、ニューモシスチス肺炎の予防若しくは治療における使用のための請求項1に記載の化合物またはその塩。
- 請求項1に記載の化合物またはその塩の有効量を患者に投与することからなる白癬症、原虫感染症、及び/又は、ニューモシスチス肺炎の予防若しくは治療方法。
- 受託番号がNITE BP-735であるカプノディアセアエ ストレインNo.339855(Capnodiaceae strain No.339855)もしくはその変異株。
- 請求項1に記載の化合物の産生能を有するカプノディアセアエ科(Capnodiaceae)の微生物を培養し、その培養物から請求項1の化合物又はその塩を単離することを含む、請求項1に記載の化合物またはその塩の製造方法。
- 微生物が請求項8記載の微生物である請求項9記載の製造方法。
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CA2761595A CA2761595A1 (en) | 2009-05-14 | 2010-05-13 | Polyketide compound |
US13/320,347 US20120059038A1 (en) | 2009-05-14 | 2010-05-13 | Polyketide compound |
CN2010800210866A CN102421909A (zh) | 2009-05-14 | 2010-05-13 | 聚酮化合物 |
AU2010248415A AU2010248415A1 (en) | 2009-05-14 | 2010-05-13 | Polyketide compound |
JP2011513364A JPWO2010131691A1 (ja) | 2009-05-14 | 2010-05-13 | ポリケチド化合物 |
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CN111436461A (zh) * | 2020-03-25 | 2020-07-24 | 福建省农业科学院茶叶研究所 | 一种生物除草剂 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
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JPS53121767A (en) | 1977-04-01 | 1978-10-24 | Chugai Pharmaceut Co Ltd | Funiculosin derivatives |
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AUPO371596A0 (en) * | 1996-11-19 | 1996-12-12 | Fujisawa Pharmaceutical Co., Ltd. | New compound |
CN1240713C (zh) * | 1998-02-09 | 2006-02-08 | 藤泽药品工业株式会社 | 化合物 |
AUPS044102A0 (en) * | 2002-02-11 | 2002-03-07 | Fujisawa Pharmaceutical Co., Ltd. | New compound |
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- 2010-05-13 CA CA2761595A patent/CA2761595A1/en not_active Abandoned
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JPS53121767A (en) | 1977-04-01 | 1978-10-24 | Chugai Pharmaceut Co Ltd | Funiculosin derivatives |
Non-Patent Citations (14)
Title |
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"Japan Collection of Microorganisms", 1995, INSTITUTE OF PHYSICAL AND CHEMICAL RESEARCH, pages: 617 |
ACTA CRYSTALLOGRAPHICA, SECTION C: CRYSTAL STRUCTURE COMMUNICATIONS, vol. C46, no. 3, 1990, pages 515 - 517 |
ANDO K. ET AL.: "Funiculosin, a new antibiotic. II. Structure elucidation and antifungal activity", JOURNAL OF ANTIBIOTICS, vol. 31, no. 6, June 1978 (1978-06-01), pages 533 - 538 * |
HIKARU KUME, TOSHIKAZU YAMAZAKI, CLINICAL MICROBIOLOGY, vol. 21, no. 5, 1994, pages 573 - 580 |
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MYCOSES, vol. 46, 2003, pages 505 - 510 |
NAWATA Y. ET AL.: "Structure of tetrahydrofuniculosin", ACTA CRYSTALLOGRAPHICA, SECTION C: CRYSTAL STRUCTURE COMMUNICATIONS, vol. 46, no. 3, March 1990 (1990-03-01), pages 515 - 517, XP009162048, DOI: doi:10.1107/S0108270189012175 * |
See also references of EP2431477A4 * |
TETRAHEDRON LETTERS, vol. 17, 1976, pages 3827 - 3830 |
TETRAHEDRON LETTERS, vol. 41, 2000, pages 9397 - 9402 |
TETRAHEDRON LETTERS, vol. 46, 2005, pages 8009 - 8010 |
THE JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 279, 2004, pages 8708 - 8714 |
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JP2015091785A (ja) * | 2013-09-30 | 2015-05-14 | 国立大学法人大阪大学 | トール様受容体4活性化作用を有するフニクロシン誘導体及びその用途 |
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EP2431477A1 (en) | 2012-03-21 |
KR20120027341A (ko) | 2012-03-21 |
CA2761595A1 (en) | 2010-11-18 |
CN102421909A (zh) | 2012-04-18 |
AU2010248415A1 (en) | 2011-12-01 |
EP2431477A4 (en) | 2012-09-26 |
US20120059038A1 (en) | 2012-03-08 |
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