WO2013065383A1 - 新規化合物、その製造方法、及びその用途 - Google Patents
新規化合物、その製造方法、及びその用途 Download PDFInfo
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- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/22—Cyclohexane rings, substituted by nitrogen atoms
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
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- 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
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- 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
- C12P17/02—Oxygen as only ring hetero atoms
- C12P17/06—Oxygen as only ring hetero atoms containing a six-membered hetero ring, e.g. fluorescein
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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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
- C12P19/46—Preparation of O-glycosides, e.g. glucosides having an oxygen atom of the saccharide radical bound to a cyclohexyl radical, e.g. kasugamycin
- C12P19/48—Preparation of O-glycosides, e.g. glucosides having an oxygen atom of the saccharide radical bound to a cyclohexyl radical, e.g. kasugamycin the cyclohexyl radical being substituted by two or more nitrogen atoms, e.g. destomycin, neamin
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- 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
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
Definitions
- the present invention includes a novel compound having a prostaglandin production inhibitory effect and a method for producing the same, a novel microorganism that is a bacterium producing the novel compound, a compound-containing composition containing the novel compound, and the compound-containing composition
- the present invention relates to a prostaglandin production inhibitor.
- Prostaglandin is a physiologically active substance that is a metabolite of arachidonic acid released from the cell membrane, and PGE 2 , PGD 2 , PGF 2 ⁇ , PGI 2 and the like are known.
- PG Prostaglandin
- PGE 2 , PGD 2 , PGF 2 ⁇ , PGI 2 and the like are known.
- the prostaglandins When a living body receives a stimulus such as a physical stimulus or an inflammatory stimulus, the prostaglandins are produced in the living body, and each of them binds to a specific receptor and causes various physiological reactions in the living body.
- the physiological reaction examples include pruritus, fever, increased vascular permeability, inflammatory reaction such as pain, abnormal increase in sensory nerve, bronchial smooth muscle contraction, platelet aggregation, tumor cell proliferation, promotion of bone resorption, nerve cell degeneration, etc.
- the prostaglandin plays an important role in the development of symptoms or pathogenesis in many diseases such as asthma, cardiovascular disease, premature birth, nephritis, atherosclerosis, overactive bladder, rheumatoid arthritis, osteoarthritis, and cancer. Is responsible. Therefore, it is expected that the various diseases can be prevented or treated by suppressing the production of the prostaglandins.
- the present invention has an excellent prostaglandin production inhibitory action, can be used for the prevention or treatment of various diseases caused by prostaglandins, and has a high safety and a method for producing the same, and the novel compounds It is an object of the present invention to provide a novel microorganism that is a production bacterium of the above, a compound-containing composition containing the novel compound, and a prostaglandin production inhibitor containing the compound-containing composition.
- the present inventors have intensively studied, and as a result, separated microorganisms from various soils, repeated research on metabolites produced by them, and belonged to the newly separated genus Saccharotrix. It was found that microorganisms produced a substance exhibiting a prostaglandin production inhibitory action in the culture solution.
- the active ingredient was separated and purified from the culture broth, and its physicochemical properties were examined.
- the obtained active ingredient was a compound represented by the following structural formula (A), which is different from any known substance, and prostaglandin. It was found that it has a gin production inhibitory action, and the present invention has been completed.
- This invention is based on the said knowledge by the present inventors, and the compound or its salt as a means for solving the said subject is a compound or its salt represented by following Structural formula (A).
- the said various problems in the past can be solved, the said objective can be achieved, it has the outstanding prostaglandin production inhibitory effect, and it uses for the prevention or treatment of the various diseases resulting from a prostaglandin.
- a highly safe novel compound and method for producing the same a novel microorganism that is a bacterium producing the novel compound, a compound-containing composition containing the novel compound, and a prostaglandin containing the compound-containing composition A production inhibitor can be provided.
- FIG. 1 is an infrared absorption spectrum obtained by measuring the compound of the present invention by the KBr method.
- FIG. 2 is an ultraviolet absorption spectrum obtained by measuring a methanol solution of the compound of the present invention.
- FIG. 3 is a proton nuclear magnetic resonance spectrum obtained by measuring a heavy dimethyl sulfoxide (heavy DMSO) solution of the compound of the present invention.
- FIG. 4 is a carbon-13 nuclear magnetic resonance spectrum obtained by measuring a heavy DMSO solution of the compound of the present invention.
- FIG. 5 is a diagram showing the prostaglandin production inhibitory action of the compound of the present invention. Vertical axis: PGE 2 or 6-keto-PFG 1 ⁇ production rate (%), horizontal axis: Compound concentration (ng / mL)
- the compound of the present invention is a compound represented by the following structural formula (A) and is a novel compound separated by the present inventors.
- the compound represented by the following structural formula (A) is a novel substance that is clearly distinguished from known compounds by the following physicochemical properties and structural characteristics.
- the ultraviolet absorption spectrum measured with a methanol solution is as shown in FIG. ⁇ max nm ( ⁇ ): 263 (9,909), 470 (2,739) (7)
- the proton nuclear magnetic resonance (NMR) spectrum measured at 25 ° C. in a heavy DMSO solvent at 600 MHz is as shown in FIG. (8)
- As a carbon 13 nuclear magnetic resonance spectrum, the carbon 13 nuclear magnetic resonance spectrum measured at 25 ° C. in heavy DMSO at 150 MHz is as shown in FIG.
- the compound has a structure represented by the structural formula (A) can be confirmed by various analysis methods selected as appropriate. For example, the mass spectrometry, the infrared absorption spectrum, the ultraviolet absorption It can be confirmed by analyzing the spectrum, the proton nuclear magnetic resonance spectrum, the carbon 13 nuclear magnetic resonance spectrum, and the like.
- the novel compound of the present invention may be a salt of the compound represented by the structural formula (A).
- the salt is not particularly limited as long as it is a pharmacologically acceptable salt, and can be appropriately selected according to the purpose. Examples thereof include organic salts such as acetates and citrates, hydrochlorides and carbonates. Examples include salt.
- the compound represented by the structural formula (A) may be obtained from a microorganism that produces the compound represented by the structural formula (A), or may be obtained by chemical synthesis. However, it is preferably obtained by the method for producing the compound of the present invention described later.
- the compound represented by the structural formula (A) has an excellent prostaglandin production inhibitory action and is a highly safe compound. Therefore, the compound represented by the structural formula (A) can be suitably used as an active ingredient such as the compound-containing composition of the present invention described later and the prostaglandin production inhibitor of the present invention.
- the method for producing the compound of the present invention is a method for producing the compound represented by the structural formula (A), and includes at least a culturing step and a collecting step, and further includes other steps as necessary.
- the culturing step is a step of culturing a microorganism belonging to the genus Saccharothrix and having an ability to produce the compound represented by the structural formula (A).
- the microorganism is not particularly limited as long as it belongs to the genus Saccharotrix and has the ability to produce the compound represented by the structural formula (A), and can be appropriately selected according to the purpose.
- the ability of the microorganism to produce the compound represented by the structural formula (A) is, for example, measuring the prostaglandin production inhibitory action of a component of the microorganism culture, preferably the culture supernatant. Examples thereof include a method and a method for detecting the compound represented by the structural formula (A) by various analysis methods.
- the microorganism culture has a prostaglandin production inhibitory action
- the microorganism has the ability to produce the compound represented by the structural formula (A). It can be determined.
- the above culture is added to cultured cells such as humans, and prostaglandins produced in the culture (for example, prostaglandin E 2 and prostaglandin I 2 stabilized metabolites). 6-keto-prostaglandin F 1 ⁇ etc.) is detected, and the culture of the microorganism in which the production of the prostaglandin is suppressed can be judged to have a prostaglandin production inhibitory effect.
- microorganism examples include the Saccharotrix sp. MI559-46F5 strain (accession number NITE P-1152) isolated by the present inventors.
- other strains capable of producing the compound represented by the structural formula (A) can be isolated from the natural world by a conventional method.
- the production bacteria that produce the compound represented by the structural formula (A) including the Saccharothrix sp. MI559-46F5 strain are subjected to irradiation and other mutation treatments, so that It is also possible to increase the productivity of the compound represented by the structural formula (A).
- production of the compound represented by the structural formula (A) by genetic engineering techniques is also possible.
- a producing bacterium that produces the compound represented by the structural formula (A) may be referred to as a nutrient medium (hereinafter simply referred to as “medium”). Inoculation) and culturing at a good temperature for the production of the compound represented by the structural formula (A).
- cultivation of actinomycetes can be used, even if it is a liquid medium. It may be an agar medium.
- the nutrient source added to the nutrient medium is not particularly limited and can be appropriately selected according to the purpose.
- a nitrogen source commercially available soybean flour, peptone, yeast extract, meat extract, corn Steep liquor, ammonium sulfate, etc. can be used, and carbohydrates such as tomato paste, glycerin, starch, glucose, galactose, dextrin, bacto-soyton, fat, etc.
- inorganic salts such as sodium chloride and calcium carbonate can be added to the medium for use, and in addition, a trace amount of metal salt can be added to the medium for use. Any of these materials may be used as long as they are useful for the production of the compound represented by the structural formula (A) by the compound-producing bacteria, and all known culture materials can be used.
- the culture method is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably cultured under aerobic conditions.
- the temperature of the culture is not particularly limited as long as the compound represented by the structural formula (A) can be produced without substantially inhibiting the growth of the compound-producing bacteria. Although it can be appropriately selected depending on the producing bacteria, it is preferably 25 ° C to 35 ° C.
- the collecting step is a step of collecting the compound represented by the structural formula (A) from the culture obtained in the culturing step. Since the compound represented by the structural formula (A) has the physicochemical properties described above, it can be collected from the culture according to its properties.
- collecting means separating and / or purifying the compound represented by the structural formula (A) from the culture.
- the culture is not particularly limited as long as it contains the compound represented by the structural formula (A) obtained in the culture step, and can be appropriately selected according to the purpose.
- Examples include culture supernatants and mixtures thereof. Among these, it is preferable to use a culture supernatant as the culture because the compound represented by the structural formula (A) can be obtained efficiently.
- the compound represented by the structural formula (A) is obtained from the cells by an extraction method using an appropriate organic solvent or an elution method by disrupting the cells. It may be extracted and subjected to separation and / or purification.
- the collection method is not particularly limited, and a method used for collecting a metabolite produced by a microorganism can be appropriately selected. Examples thereof include a solvent extraction method, a method using a difference in adsorption affinity for various adsorbents, and a chromatography method.
- the compounds represented by the structural formula (A) can be separated and / or purified by using these methods alone or in combination as appropriate, and repeatedly using them in some cases.
- the solvent used in the solvent extraction method is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include ethyl acetate and n-butanol.
- adsorbent there is no restriction
- the chromatographic method is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include thin layer chromatography, high performance liquid chromatography for fractionation using normal phase or reverse phase columns (preparation). HPLC) method and the like. There is no restriction
- the method for eluting the compound represented by the structural formula (A) from the adsorbent or the carrier in the chromatography is not particularly limited, and is appropriately selected according to the kind and properties of the adsorbent and the carrier. be able to.
- a method of elution using water-containing alcohol, water-containing acetone or the like as an elution solvent can be mentioned.
- the compound represented by the structural formula (A) can be produced, whereby the compound represented by the structural formula (A) can be suitably obtained.
- the microorganism of the present invention belongs to the genus Saccharothrix and has the ability to produce the compound represented by the structural formula (A).
- the microorganism has the ability to produce the compound represented by the structural formula (A). Therefore, in the method for producing the compound of the present invention, the microorganism producing the compound represented by the structural formula (A) is used. If it is microorganisms which can be used as, there will be no restriction
- microorganisms it is particularly preferable to use a microorganism having a strain number of MI559-46F5 strain.
- the MI559-46F5 strain was registered on September 28, 2011 at the Patent Microorganism Depositary Center (National Institute of Technology and Evaluation 2-5-8, Kazusa Kamashichi, Kisarazu City, Chiba Prefecture) on September 28, 2011. Commissioned as NITE P-1152.
- the above-mentioned MI559-46F5 strain is susceptible to changes in properties.
- mutant strains derived from the MI559-46F5 strain for example, ultraviolet rays, X-rays, radiation, drugs, etc.
- a zygote, a genetic recombinant, etc. that can be obtained by the mutation treatment of the above, those having the ability to produce the compound represented by the structural formula (A) are Included in the microorganism of the invention.
- Compound-containing composition contains at least one of the compound represented by the structural formula (A) and a salt thereof, and further contains other components as necessary.
- the compound-containing composition may be the compound itself represented by the structural formula (A).
- ⁇ Other ingredients> There is no restriction
- an additive, an adjuvant, water, etc. are mentioned. These may be used alone or in combination of two or more.
- the additive or the adjuvant is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include bactericides, preservatives, binders, thickeners, fixing agents, binders, and coloring agents. , Stabilizers, pH adjusters, buffers, isotonic agents, solvents, antioxidants, UV inhibitors, crystal precipitation inhibitors, antifoaming agents, physical property improvers, preservatives, and the like.
- the bactericide is not particularly limited and may be appropriately selected depending on the intended purpose.
- examples thereof include cationic surfactants such as benzalkonium chloride, benzethonium chloride and cetylpyridinium chloride.
- the preservative is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include p-hydroxybenzoates, chlorobutanol, and cresol.
- the binder, thickener and fixing agent are not particularly limited and may be appropriately selected depending on the intended purpose.
- the binder is not particularly limited and may be appropriately selected depending on the intended purpose.
- examples include propyl starch, methyl cellulose, ethyl cellulose, shellac, calcium phosphate, and polyvinyl pyrrolidone.
- the colorant is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include titanium oxide and iron oxide.
- the stabilizer is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include tragacanth, gum arabic, gelatin, sodium pyrosulfite, EDTA, thioglycolic acid, and thiolactic acid.
- the pH adjusting agent and the buffering agent are not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include sodium citrate, sodium acetate, and sodium phosphate.
- the tonicity agent is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include sodium chloride and glucose.
- the compound-containing composition contains at least one of the compound represented by the structural formula (A) and a salt thereof, the compound-containing composition has an excellent prostaglandin production inhibitory action and is highly safe.
- the prostaglandin production inhibitor of the present invention can be suitably used.
- the prostaglandin production inhibitor of the present invention contains the compound-containing composition of the present invention, and further contains other components as necessary.
- the prostaglandin production inhibitor has a prostaglandin production inhibitory action.
- the prostaglandin production inhibitor may be the compound-containing composition itself.
- the said prostaglandin production inhibitor may be used individually by 1 type, and may be used with the pharmaceutical which uses another component as an active ingredient. Moreover, you may use the said prostaglandin production inhibitor in the state mix
- the method for examining the prostaglandin production inhibitory activity is not particularly limited and may be appropriately selected from known methods according to the purpose. Examples thereof include the methods described in Test Examples described later.
- ⁇ Dosage form> There is no restriction
- the solid agent is not particularly limited and may be appropriately selected depending on the intended purpose.
- a content agent for example, tablet, chewable tablet, effervescent tablet, orally disintegrating tablet, troche, drop agent , Hard capsules, soft capsules, granules, powders, pills, dry syrups, soaking agents and the like.
- a suppository, a poultice, a plaster agent etc. are mentioned, for example.
- liquid agent a syrup agent, a drink agent, a suspension agent, an alcoholic agent etc. are mentioned, for example.
- a liquid agent, eye drops, an aerosol agent, a spray agent etc. are mentioned, for example.
- administering There is no restriction
- limiting in particular as said administration method According to the objective, it can select suitably, For example, a local administration method, an enteral administration method, a parenteral administration method etc. are mentioned.
- the dosage is not particularly limited and may be appropriately selected in consideration of various factors such as the age, weight, constitution, symptom, and presence / absence of administration of a drug containing other ingredients as active ingredients. it can.
- the animal species to be administered is not particularly limited and can be appropriately selected according to the purpose. For example, human, monkey, pig, cow, sheep, goat, dog, cat, mouse, rat, bird, etc. Among these, it is preferably used for humans.
- the prostaglandin production inhibitor has an excellent prostaglandin production inhibitory action and is highly safe. Therefore, pruritus caused by prostaglandin, fever, increased vascular permeability, pain and other inflammatory reactions, sensory nerves Can suppress physiological activities such as abnormal hypersensitivity, bronchial smooth muscle contraction, platelet aggregation, tumor cell growth, bone resorption promotion, neuronal degeneration, asthma, cardiovascular disease, premature birth, nephritis, atherosclerosis, overactivity It can be suitably used as a preventive or therapeutic agent for various diseases such as bladder, rheumatoid arthritis, osteoarthritis, and cancer.
- MI559-46F5 strain (deposited under the accession number NITE P-1152) pre-cultured on an agar slant medium, and 2 at 30 ° C. By culturing with shaking at a rotational speed of 180 rpm for one day, a seed culture solution was obtained.
- glycerin 2.0%, dextrin 2.0%, yeast extract (manufactured by Wako Pure Chemical Industries, Ltd.) 0.3%, bacto soyton (manufactured by Difco) 1.0%, ammonium sulfate 0.2%, and carbonic acid Calcium 0.2% was suspended in water to prepare a production medium liquid medium (adjusted to pH 7.0).
- 110 mL of this liquid medium for production medium was dispensed into Erlenmeyer flasks (500 mL volume) and sterilized at 120 ° C. for 20 minutes by a conventional method.
- the liquid medium for production medium was inoculated with 2% by volume of the prepared seed culture, and cultured with shaking at 27 ° C. for 5 days at a rotation speed of 180 rpm.
- the culture solution 15L obtained in the culturing step was centrifuged at 8,000 rpm for 15 minutes to separate the culture filtrate and the cells. Subsequently, the obtained culture was applied to a column (inner diameter: 80 mm ⁇ length: 300 mm) packed with 1.5 L of an adsorption resin (Diaion (registered trademark) HP-20, manufactured by Mitsubishi Chemical Corporation) equilibrated with water. After passing the filtrate, the adsorbent resin was washed with 3 L of water, followed by 4.5 L of 50% by volume methanol aqueous solution.
- adsorption resin Diaion (registered trademark) HP-20, manufactured by Mitsubishi Chemical Corporation
- the washed adsorbent resin was eluted with 3 L of methanol, methanol was distilled off from the eluate with an evaporator, and the resulting residue was dissolved in 1.5 L of water.
- the mixture was stirred by adding 1.5 L of ethyl acetate, allowed to stand to separate into two phases, and collecting the ethyl acetate phase. This washing operation was further continued twice, and then ethyl acetate was distilled off with an evaporator to obtain 1.13 g of a red oily substance.
- the obtained red oily substance was dissolved in a small amount of methanol, applied to celite, and applied to a 140 mL silica gel column packed with chloroform. Wash with 0.8 L of chloroform, then wash with 0.8 L of chloroform-methanol mixture (100: 1 (volume ratio)), and further 1.2 L of chloroform-methanol mixture (100: 2 (volume ratio)) And then eluted with 1.2 L of a chloroform-methanol mixture (10: 1 (volume ratio)). The eluate was concentrated under reduced pressure to obtain 127.6 mg of a red oily substance.
- the red oily substance was dissolved in methanol, applied to a column (Toyopearl HW-40F, inner diameter 37 mm ⁇ length 670 mm, manufactured by Tosoh Corporation), and eluted with methanol. The eluate was collected and concentrated under reduced pressure to obtain a reddish purple oily substance.
- SW982 cells purchased from American Type Culture Collection: ATCC
- a human osteosarcoma cell line containing 10% fetal bovine serum (manufactured by Nichirei) (DMEM / F12) Modified Eagle Medium: The suspension was suspended in Nutrient Mixture F-12 (manufactured by Invitrogen), seeded in a 96-well culture plate at 1 ⁇ 10 4 cells / well, and cultured overnight.
- Nutrient Mixture F-12 manufactured by Invitrogen
- HTRF time-resolved fluorescence
- PGE 2 production rate (%) (AB) / (CB) ⁇ 100
- A represents the PGE 2 concentration in the test substance addition system
- B represents the PGE 2 concentration in the negative control
- C represents the PGE 2 concentration in the positive control .
- PGI 2 production inhibitory action The PGI 2 production inhibitory action of the compound represented by the structural formula (A) was examined by the following method. PGI 2 is metabolized to 6-keto-PGF 1 ⁇ (6-keto-prostaglandin F 1 ⁇ ), which is a stable metabolite. Therefore, by measuring the 6-keto-PGF 1 ⁇ concentration, the concentration of PGI 2 produced can be measured.
- SW982 cells SW982 cells (purchased from ATCC), a human osteosarcoma cell line, are suspended in DMEM / F12 (Invitrogen) containing 10% fetal bovine serum (Biowest), and then 1 ⁇ 10 4 in a 96-well culture plate. Cells / well were seeded and cultured overnight.
- the culture supernatant was collected, and the 6-keto-PGF 1 ⁇ concentration D in the culture supernatant was measured using a 6-keto prostaglandin F1 ⁇ enzyme immunoassay kit (# 515211, manufactured by CAYMAN). .
- 6-keto-PGF 1 ⁇ concentration in negative control As a negative control, in the measurement of the 6-keto-PGF 1 ⁇ concentration in the test substance addition system, the test substance and bradykinin (stimulant) were not added, except that 6-keto-PGF in the test substance addition system was not added. The same operation as the measurement of 1 ⁇ concentration was performed, and 6-keto-PGF 1 ⁇ concentration E was measured.
- 6-keto-PGF 1 ⁇ concentration in positive control The positive control was the same as the measurement of 6-keto-PGF 1 ⁇ concentration in the test substance addition system except that no test substance was added in the measurement of 6-keto-PGF 1 ⁇ concentration in the test substance addition system.
- the 6-keto-PGF 1 ⁇ concentration F was measured.
- 6-keto-PGF 1 ⁇ production rate (DE) / (FE) ⁇ 100
- D represents the 6-keto-PGF 1 ⁇ concentration in the test substance addition system
- E represents the 6-keto-PGF 1 ⁇ concentration in the negative control
- F represents Represents 6-keto-PGF 1 ⁇ concentration in the positive control.
- Test Example 3 when the test substance contains a substance effective for suppressing prostaglandin production, the production of PGI 2 in the cell culture medium is suppressed, and thus the stability of PGI 2 detected from the culture liquid The metabolite 6-keto-PGF 1 ⁇ is reduced. That is, the 6-keto-PGF 1 ⁇ production rate decreases.
- the compound represented by the structural formula (A) As a result of adding the compound represented by the structural formula (A) as a test substance, the production rate of 6-keto-PGF 1 ⁇ decreased as shown in FIG. Therefore, it was confirmed that the compound represented by the structural formula (A) has an excellent prostaglandin production inhibitory action.
- test substance addition system- SW982 cells purchased from ATCC
- SW982 cells purchased from ATCC
- DMEM / F12 medium Invitrogen
- 10% fetal calf serum Naichirei Co., Ltd.
- 96 wells Each 0.1 mL of the culture plate was inoculated.
- the test substance (compound represented by the structural formula (A)) produced in Production Example 1 was added to each well so as to have the concentration shown in Table 1 below, and the test substance was added at 37 ° C., 5% CO 2. For 48 hours.
- the compound represented by the structural formula (A) does not affect cell proliferation with respect to SW982 cells. It was confirmed that the safety was high.
- Examples of the aspect of the present invention include the following. ⁇ 1> A compound represented by the following structural formula (A) or a salt thereof. ⁇ 2> A method for producing a compound represented by the following structural formula (A), A culture step of culturing a microorganism belonging to the genus Saccharothrix and having the ability to produce a compound represented by the following structural formula (A): A collecting step of collecting a compound represented by the following structural formula (A) from the culture obtained in the culturing step; It is a manufacturing method of the compound characterized by including. ⁇ 3> A microorganism belonging to the genus Saccharothrix and capable of producing the compound represented by the structural formula (A) is Saccharothris sp.
- a prostaglandin production inhibitor comprising the compound-containing composition according to ⁇ 6> and having a prostaglandin production inhibitory action.
- the compound of the present invention has an excellent prostaglandin production inhibitory action and is a highly safe compound. Therefore, pruritus caused by prostaglandins, fever, increased vascular permeability, pain and other inflammatory reactions, sensory nerves Can suppress physiological activities such as abnormal hypersensitivity, bronchial smooth muscle contraction, platelet aggregation, tumor cell growth, bone resorption promotion, neuronal degeneration, asthma, cardiovascular disease, premature birth, nephritis, atherosclerosis, overactivity It can be suitably used as an active ingredient of preventive or therapeutic agents for various diseases such as bladder, rheumatoid arthritis, osteoarthritis, and cancer.
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Abstract
Description
したがって、前記プロスタグランジンの産生を抑制することにより、前記種々の疾患を予防又は治療できることが期待されている。
本発明の化合物は、下記構造式(A)で表される化合物であり、本発明者らが分離した新規化合物である。
下記構造式(A)で表される化合物は、下記に示す物理化学的性質及び構造上の特徴によって、既知の化合物と明確に区別される新規物質である。
前記構造式(A)で表される化合物の物理化学的性質としては、次のとおりである。
(1) 外観は、赤紫色の粉状である。
(2) 分子式は、C20H24N2O9で表される。
(3) 高分解能質量分析(HRESIMS:正イオンモード)による、実験値は、m/z 459.1371(M+Na)+であり、計算値は、m/z 459.1374(C20H24N2O9Naとして)である。
(4) 比旋光度は、[α]D23=-358°(c 0.0036, メタノール)である。
(5) KBr法で測定した赤外吸収スペクトルは、図1に示すとおりである。
(6) メタノール溶液で測定した紫外吸収スペクトルは、図2に示すとおりである。
λmax nm(ε) :263(9,909)、470(2,739)
(7) プロトン核磁気共鳴(NMR)スペクトルとして、600MHzにおいて重DMSO溶媒中で25℃にて測定したプロトン核磁気共鳴スペクトルは、図3に示すとおりである。
(8) 炭素13核磁気共鳴スペクトルとして、150MHzにおいて重DMSO中で25℃にて測定した炭素13核磁気共鳴スペクトルは、図4に示すとおりである。
前記塩としては、薬理学的に許容され得る塩であれば、特に制限はなく、目的に応じて適宜選択することができ、例えば、酢酸塩、クエン酸塩等の有機塩、塩酸塩、炭酸塩などが挙げられる。
前記構造式(A)で表される化合物は、優れたプロスタグランジン産生抑制作用を有し、安全性の高い化合物である。そのため、前記構造式(A)で表される化合物は、例えば、後述する本発明の化合物含有組成物や、本発明のプロスタグランジン産生抑制剤等の有効成分として好適に利用可能である。
本発明の化合物の製造方法は、前記構造式(A)で表される化合物の製造方法であって、培養工程と、採取工程と、を少なくとも含み、必要に応じて更にその他の工程を含む。
前記培養工程は、サッカロスリックス(Saccharothrix)属に属し、前記構造式(A)で表される化合物を生産する能力を有する微生物を培養する工程である。
具体的には、ヒト等の培養細胞に前記培養物を添加し、該培養物中に産生されるプロスタグランジン(例えば、プロスタグランジンE2やプロスタグランジンI2の安定化代謝物である6-ケト-プロスタグランジンF1αなど)を検出し、前記プロスタグランジンの産生が抑制された微生物の培養物をプロスタグランジン産生抑制作用を有すると判断することができる。
前記栄養培地に添加する栄養源としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、窒素源として、市販されている大豆粉、ペプトン、酵母エキス、肉エキス、コーン・スティープ・リカー、硫酸アンモニウムなどが使用でき、炭素源として、トマトペースト、グリセリン、でん粉、グルコース、ガラクトース、デキストリン、バクトソイトン等の炭水化物、脂肪などが使用できる。更に、食塩、炭酸カルシウム等の無機塩を培地に添加して使用することもでき、その他、必要に応じて微量の金属塩を培地に添加して使用することもできる。
これらの材料は、前記化合物類生産菌が利用し、前記構造式(A)で表される化合物の生産に役立つものであればよく、公知の培養材料は全て用いることができる。
前記培養の温度としては、前記化合物類生産菌の発育が実質的に阻害されずに、前記構造式(A)で表される化合物を生産しうる範囲であれば、特に制限はなく、使用する生産菌に応じて適宜選択することができるが、25℃~35℃が好ましい。
前記培養の期間としては、特に制限はなく、前記構造式(A)で表される化合物の蓄積に合わせて適宜選択することができる。
前記採取工程は、前記培養工程で得られた培養物から前記構造式(A)で表される化合物を採取する工程である。前記構造式(A)で表される化合物は、上述した物理化学的性質を有するので、その性状に従って培養物から採取することができる。ここで、本発明において、採取とは、前記構造式(A)で表される化合物を、前記培養物から分離及び/又は精製することを意味する。
なお、前記培養物として、前記菌体を用いる場合は、適当な有機溶媒を用いた抽出方法や、菌体破砕による溶出方法などにより、前記構造式(A)で表される化合物を菌体から抽出し、これを分離及び/又は精製に供してもよい。
前記吸着剤の市販品の具体例としては、アンバーライトXAD(ローム・アンド・ハース社製)、ダイヤイオンHP-20(三菱化学株式会社製)などが挙げられる。
前記クロマトグラフィー法に用いる担体としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、ゲル濾過、シリカゲル、アルミナ、活性炭などが挙げられる。
前記ゲル濾過クロマトグラフィー法に用いる担体の市販品の具体例としては、トヨパールHW-40F(東ソー株式会社製)、セファデックス(Sephadex)LH-20(GE社製)などが挙げられる。
前記その他の工程としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、前記培養工程で得られた培養物又は前記採取工程で得られた前記構造式(A)で表される化合物を洗浄する洗浄工程、前記採取工程で得られた前記構造式(A)で表される化合物を更に精製する精製工程などが挙げられる。前記洗浄工程や前記精製工程は、公知の方法で適宜行われる。
本発明の微生物は、サッカロスリックス(Saccharothrix)属に属し、前記構造式(A)で表される化合物を生産する能力を有する。前記微生物は、前記構造式(A)で表される化合物を生産する能力を有し、そのために、本発明の前記化合物の製造方法において、前記構造式(A)で表される化合物の生産菌として使用され得る微生物であれば、特に制限はなく、目的に応じて適宜選択することができる。
本発明の化合物含有組成物は、少なくとも前記構造式(A)で表される化合物及びその塩の少なくともいずれかを含み、必要に応じて更にその他の成分を含む。
前記化合物含有組成物中の前記構造式(A)で表される化合物の含有量としては、特に制限はなく、目的に応じて適宜選択することができる。また、前記化合物含有組成物は、前記構造式(A)で表される化合物そのものであってもよい。
前記その他の成分としては、特に制限はなく、例えば、薬理学的に許容され得る担体の中から目的に応じて適宜選択することができ、例えば、添加剤、補助剤、水などが挙げられる。これらは、1種単独で使用してもよく、2種以上を併用してもよい。
前記添加剤又は前記補助剤としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、殺菌剤、保存剤、粘結剤、増粘剤、固着剤、結合剤、着色剤、安定化剤、pH調整剤、緩衝剤、等張化剤、溶剤、酸化防止剤、紫外線防止剤、結晶析出防止剤、消泡剤、物性向上剤、防腐剤などが挙げられる。
前記化合物含有組成物は、前記構造式(A)で表される化合物及びその塩の少なくともいずれかを含むため、優れたプロスタグランジン産生抑制作用を有し、安全性の高いものであり、後述する本発明のプロスタグランジン産生抑制剤などに好適に利用可能である。
本発明のプロスタグランジン産生抑制剤は、本発明の前記化合物含有組成物を含有し、必要に応じて、更にその他の成分を含有する。前記プロスタグランジン産生抑制剤は、プロスタグランジン産生抑制作用を有する。
前記プロスタグランジン産生抑制剤中の前記化合物含有組成物の含有量としては、特に制限はなく、目的に応じて適宜選択することができる。また、前記プロスタグランジン産生抑制剤は、前記化合物含有組成物そのものであってもよい。
前記その他の成分としては、特に制限はなく、例えば、前記化合物含有組成物中のその他の成分と同様のものなどが挙げられる。
前記プロスタグランジン産生抑制剤中の、前記その他の成分の含有量としては、特に制限はなく、前記構造式(A)で表される化合物の効果を損なわない範囲内で、目的に応じて適宜選択することができる。
前記プロスタグランジン産生抑制活性を調べる方法としては、特に制限はなく、公知の方法の中から目的に応じて適宜選択することができ、例えば、後述する試験例に記載の方法などが挙げられる。
前記プロスタグランジン産生抑制剤の剤型としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、固形剤、半固形剤、液剤などが挙げられる。
前記固形剤としては、特に制限はなく、目的に応じて適宜選択することができるが、内容剤として用いられる場合、例えば、錠剤、チュアブル錠、発泡錠、口腔内崩壊錠、トローチ剤、ドロップ剤、硬カプセル剤、軟カプセル剤、顆粒剤、散剤、丸剤、ドライシロップ剤、浸剤などが挙げられる。
前記固形剤が、外用剤として用いられる場合、例えば、坐剤、パップ剤、プラスター剤などが挙げられる。
前記半固形剤としては、特に制限はなく、目的に応じて適宜選択することができるが、内用剤として用いられる場合、例えば、舐剤、チューインガム剤、ホイップ剤、ゼリー剤などが挙げられる。
前記半固形剤が、外用剤として用いられる場合、例えば、軟膏剤、クリーム剤、ムース剤、インヘラー剤、ナザールジェル剤などが挙げられる。
前記液剤としては、特に制限はなく、目的に応じて適宜選択することができるが、内用剤として用いられる場合、例えば、シロップ剤、ドリンク剤、懸濁剤、酒精剤などが挙げられる。
前記液剤が、外用剤として用いられる場合、例えば、液剤、点眼剤、エアゾール剤、噴霧剤などが挙げられる。
前記プロスタグランジン産生抑制剤の投与方法、投与量、投与時期、及び投与対象としては、特に制限はなく、目的に応じて適宜選択することができる。
前記投与方法としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、局所投与法、経腸投与法、非経口投与法などが挙げられる。
前記投与量としては、特に制限はなく、投与対象個体の年齢、体重、体質、症状、他の成分を有効成分とする医薬の投与の有無など、様々な要因を考慮して適宜選択することができる。
前記投与対象となる動物種としては、特に制限はなく、目的に応じて適宜選択することができ、例えば、ヒト、サル、ブタ、ウシ、ヒツジ、ヤギ、イヌ、ネコ、マウス、ラット、トリなどが挙げられるが、これらの中でもヒトに好適に用いられる。
前記プロスタグランジン産生抑制剤は、優れたプロスタグランジン産生抑制作用を有し、安全性が高いため、プロスタグランジンに起因する掻痒、発熱、血管透過性亢進、疼痛等の炎症反応、知覚神経の異常亢進、気管支平滑筋収縮、血小板凝集、腫瘍細胞増殖、骨吸収促進、神経細胞変性等の生理活性を抑制することができ、喘息、心臓血管疾患、早産、腎炎、アテローム硬化症、過活動膀胱、慢性関節リウマチ、変形性関節炎、癌等の種々の疾患の予防薬又は治療薬として好適に利用可能である。
<培養工程>
ガラクトース2%、デキストリン2%、グリセリン1%、バクトソイトン(ディフコ社製)1%、コーン・スティープ・リカー0.5%、硫酸アンモニウム0.2%、及び炭酸カルシウム0.2%を水に懸濁し、種母培養液用液体培地(pH7.0に調整)を調製した。この種母培養液用液体培地を三角フラスコ(500mL容)に110mLずつ分注し、常法により120℃で20分滅菌した。
前記滅菌した種母培養液用液体培地に、寒天斜面培地で前培養したサッカロスリックス エスピー(Saccharothrix sp.)MI559-46F5株(受託番号NITE P-1152として寄託)を接種し、30℃で2日間、回転速度180rpmで振とう培養することで、種母培養液を得た。
前記培養工程で得られた培養液15Lを、回転数8,000rpmで15分間遠心分離し、培養ろ液と菌体とに分離した。次いで、水で平衡化した1.5Lの吸着樹脂(ダイヤイオン(登録商標)HP-20、三菱化学株式会社製)を充填したカラム(内径80mm×長さ300mm)、に、得られた前記培養ろ液を通過させた後、該吸着樹脂を水3Lで洗浄し、続いて50体積%メタノール水溶液4.5Lで洗浄した。洗浄後の吸着樹脂にメタノール3Lを供し溶出させ、この溶出液からメタノールをエバポレーターで留去し、得られた残渣を水1.5Lに溶解した。ここに酢酸エチル1.5Lを添加して攪拌し、これを静置して二相に分離させ、酢酸エチル相を回収することにより洗浄を行った。この洗浄の操作を更に続けて2回行った後、酢酸エチルをエバポレーターで留去し、赤色油状物質1.13gを得た。
製造例1で得られた目的物質の物理化学的性質を測定したところ、以下の通りであり、これらのことから、前記目的物質が、下記構造式(A)で表される構造を有する新規化合物であることが確認された。
(1) 外観は、赤紫色の粉状であった。
(2) 分子式は、C20H24N2O9で表される。
(3) 高分解能質量分析(HRESIMS:正イオンモード)による、実験値は、m/z 459.1371(M+Na)+であり、計算値は、m/z 459.1374(C20H24N2O9Naとして)であった。
(4) 比旋光度は、[α]D23=-358°(c 0.0036, メタノール)である。
(5) KBr法で測定した赤外吸収スペクトルは、図1に示すとおりであった。
(6) メタノール溶液で測定した紫外吸収スペクトルは、図2に示すとおりであった。
λmax nm(ε) :263(9,909)、470(2,739)
(7) プロトン核磁気共鳴スペクトルとして、600MHzにおいて重ジメチルスルホキシド(DMSO)溶媒中で25℃にて測定したプロトン核磁気共鳴スペクトルは、図3に示すとおりであった。
(8) 炭素13核磁気共鳴スペクトルとして、150MHzにおいて重DMSO中で25℃にて測定した炭素13核磁気共鳴スペクトルは、図4に示すとおりであった。
前記構造式(A)で表される化合物のPGE2産生抑制作用について以下の方法で検討を行った。
ヒト骨肉腫細胞株であるSW982細胞(米国の株保存機関(American Type Culture Collection:ATCC)より購入)を10%牛胎児血清(ニチレイ社製)を含むDMEM/F12(Dulbecco’s Modified Eagle Medium:Nutrient Mixture F-12、インビトロジェン社製)に懸濁した後、96ウェル培養プレートに1×104細胞/ウェルとなるように播種し、一晩培養した。
前記一晩培養後のSW982細胞における96ウェル培養プレート中の培地をHEPES-HANKSバッファー(日水製薬株式会社製)に置き換え、前記製造例1で製造した被験物質(前記構造式(A)で表される化合物)を10,000ng/mL、3,333ng/mL、1,111ng/mL、370ng/mL、123ng/mL、41ng/mL、14ng/mL、5ng/mL、又は2ng/mLとなるように各ウェルに添加し、更に炎症性の刺激剤としてブラジキニン(株式会社ペプチド研究所製)を1nMになるように添加した。30分間培養後、培養上清を回収し、該培養上清中のPGE2濃度Aを、均一時間分解蛍光(HTRF)法ベースのキット(62P2APEB、Cisbio International社製)を用いて測定した。
陰性対照としては、前記被験物質添加系におけるPGE2濃度の測定において、被験物質及びブラジキニン(刺激剤)を添加しなかったこと以外は、前記被験物質添加系におけるPGE2濃度の測定と同様の操作を行い、PGE2濃度Bの測定を行った。
陽性対照としては、前記被験物質添加系におけるPGE2濃度の測定において、被験物質を添加しなかったこと以外は、前記被験物質添加系におけるPGE2濃度の測定と同様の操作を行い、PGE2濃度Cの測定を行った。
各ウェルのPGE2濃度の測定結果より、下記式(1)に基づきPGE2産生率を算出した。
PGE2産生率(%)=(A-B)/(C-B)×100 ・・・式(1)
前記式(1)において、「A」は、被験物質添加系におけるPGE2濃度を表し、「B」は、陰性対照におけるPGE2濃度を表し、「C」は、陽性対照におけるPGE2濃度を表す。
前記構造式(A)で表される化合物を被験物質として添加した結果、図5に示すとおり、PGE2産生率が低下した。
したがって、前記構造式(A)で表される化合物が、優れたプロスタグランジン産生抑制作用を有することが認められた。
前記構造式(A)で表される化合物のPGI2産生抑制作用について、以下の方法で検討を行った。PGI2は、代謝されて安定代謝物である6-ケト-PGF1α(6-ケト-プロスタグランジンF1α)となる。したがって、6-ケト-PGF1α濃度を測定することで、産生されたPGI2濃度を測定することができる。
ヒト骨肉腫細胞株であるSW982細胞(ATCCより購入)を10%牛胎児血清(Biowest社製)を含むDMEM/F12(インビトロジェン社製)に懸濁した後、96ウェル培養プレートに1×104細胞/ウェルとなるように播種し、一晩培養した。
前記一晩培養後のSW982細胞における96ウェル培養プレート中の培地をHEPES-HANKSバッファー(日水製薬株式会社製)に置き換え、前記製造例1で製造した被験物質(前記構造式(A)で表される化合物)を10,000ng/mL、3,333ng/mL、1,111ng/mL、370ng/mL、123ng/mL、41ng/mL、又は14ng/mLとなるように各ウェルに添加し、更に炎症性の刺激剤としてブラジキニン(株式会社ペプチド研究所製)を1nMになるように添加した。30分間培養後、培養上清を回収し、該培養上清中の6-ケト-PGF1α濃度Dを、6-keto Prostaglandin F1 α enzyme immunoassay kit(#515211、CAYMAN社製)を用いて測定した。
陰性対照としては、前記被験物質添加系における6-ケト-PGF1α濃度の測定において、被験物質及びブラジキニン(刺激剤)を添加しなかったこと以外は、前記被験物質添加系における6-ケト-PGF1α濃度の測定と同様の操作を行い、6-ケト-PGF1α濃度Eの測定を行った。
陽性対照としては、前記被験物質添加系における6-ケト-PGF1α濃度の測定において、被験物質を添加しなかったこと以外は、前記被験物質添加系における6-ケト-PGF1α濃度の測定と同様の操作を行い、6-ケト-PGF1α濃度Fの測定を行った。
各ウェルの6-ケト-PGF1α濃度の測定結果より、下記式(2)に基づき6-ケト-PGF1α産生率を算出した。
6-ケト-PGF1α産生率(%)=(D-E)/(F-E)×100 ・・・式(2)
前記式(2)において、「D」は、被験物質添加系における6-ケト-PGF1α濃度を表し、「E」は、陰性対照における6-ケト-PGF1α濃度を表し、「F」は、陽性対照における6-ケト-PGF1α濃度を表す。
前記構造式(A)で表される化合物を被験物質として添加した結果、図5に示すとおり、6-ケト-PGF1α産生率が低下した。
したがって、前記構造式(A)で表される化合物が、優れたプロスタグランジン産生抑制作用を有することが認められた。
前記構造式(A)で表される化合物の細胞毒性について、以下の方法で検討を行った。
SW982細胞(ATCCより購入)を、10%胎仔牛血清(株式会社ニチレイ製)を含むDMEM/F12培地(インビトロジェン社製)を用いて2×103個/ウェルとなるように調製し、96ウェル培養プレートに0.1mLずつ接種した。これと同時に、前記製造例1で製造した被験物質(前記構造式(A)で表される化合物)を下記表1に示す濃度となるように各ウェルに添加し、37℃、5%CO2の条件下で48時間培養した。48時間培養後、各ウェルに細胞数測定キット(cell counting Kit-8、同仁化学株式会社製)10μLを添加し、更に2時間培養した後、450nmの吸光度Gを測定した。
前記細胞数測定キットは、生存細胞が発色する。したがって、450nmの吸光度が高いほど、生存細胞が多いこと、即ち細胞増殖に影響を及ぼさないことを示す。これより、各ウェルの吸光度の測定結果から、下記式(3)に基づき細胞増殖率を算出し、これにより細胞増殖に及ぼす影響を評価した。
細胞増殖率(%)=G/H×100 ・・・式(3)
前記式(3)において、「G」は、被験物質添加系における450nmの吸光度を表し、「H」は、被験物質非添加系(下記表1の濃度0nM)における450nmの吸光度を表す。
<1> 下記構造式(A)で表されることを特徴とする化合物又はその塩である。
サッカロスリックス(Saccharothrix)属に属し、下記構造式(A)で表される化合物を生産する能力を有する微生物を培養する培養工程と、
前記培養工程で得られた培養物から下記構造式(A)で表される化合物を採取する採取工程と、
を含むことを特徴とする化合物の製造方法である。
<4> サッカロスリックス(Saccharothrix)属に属し、下記構造式(A)で表される化合物を生産する能力を有することを特徴とする微生物である。
<6> 下記構造式(A)で表される化合物及びその塩の少なくともいずれかを含有することを特徴とする化合物含有組成物である。
Claims (7)
- サッカロスリックス(Saccharothrix)属に属し、構造式(A)で表される化合物を生産する能力を有する微生物が、受託番号NITE P-1152のサッカロスリックス エスピー(Saccharothrix sp.)MI559-46F5株である請求項2に記載の化合物の製造方法。
- 受託番号NITE P-1152のサッカロスリックス エスピー(Saccharothrix sp.)MI559-46F5株である請求項4に記載の微生物。
- 請求項6に記載の化合物含有組成物を含有し、プロスタグランジン産生抑制作用を有することを特徴とするプロスタグランジン産生抑制剤。
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JP2004262868A (ja) | 2003-03-03 | 2004-09-24 | Minofuaagen Seiyaku:Kk | 新規なオレアナン系トリテルペン誘導体、該誘導体の製造方法及び原料、並びに該誘導体を有効成分とする医薬組成物、抗炎症剤及びプロスタグランジンe2産生抑制剤 |
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KOICHI NAKAE ET AL.: "Hosenkin Taisha Sanbutsu kara Erareta Shinki Prostaglandin Hoshutsu Sogai Busshitsu pronqodine A ni Kansuru Kenkyu", JAPAN SOCIETY FOR BIOSCIENCE, BIOTECHNOLOGY, AND AGROCHEMISTRY 2011 NENDO TAIKAI KOEN YOSHISHU, 5 March 2011 (2011-03-05), pages 18 * |
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