JP2001247571A - Condensed quinolone derivative - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new compound having therapeutic effects on sepsis, etc., and a medicine comprising the same as an active ingredient. SOLUTION: This new quinolone compound derivative (1) having an inhibitory activity against the production of NO(nitric oxide) and TNF'-α (tumor necrosis factor) and represented by the formula (wherein R1 means 2-propyl or 2-butyl; and R2 means hydrogen atom or hydroxy, with the proviso that R2 means hydrogen atom when R1 is 2-propyl) is produced in a culture medium by culturing a microorganism belonging to the genus Penicillium in the culture medium. The medicine comprises the compound or its salt as an active ingredient and is preferably a medicine which is a prophylactic and a therapeutic agents for systemic inflammatory response syndrome, the sepsis, multiple organ failure, disseminated intravascular coagulation syndrome or endotoxin shock.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a novel condensed quinolone derivative or a salt thereof, which is a fermentation product, and a medicament containing the derivative as an active ingredient, preferably a systemic inflammatory response syndrome, sepsis, multiple organ failure, seeding. The present invention relates to a medicament that is an agent for preventing and treating sexual coagulation syndrome, endotoxin shock and the like.

[0002]

2. Description of the Related Art As effective treatment means for severe bacterial infections such as sepsis, control of bacteria causing shock, neutralization / absorption of free endotoxin, and neutralization / suppression of production of physiologically active substances such as cytokines are known. And so on. On the other hand, it is known that the use of certain antibiotics causes rapid release of endotoxin from bacterial cells.

[0003]

An object of the present invention is to provide a novel compound having a therapeutic effect on sepsis and the like and a medicament containing the same as an active ingredient.

[0004]

Under the above circumstances, the present inventors have conducted intensive studies on substances produced by many naturally occurring microorganisms. As a result, the microorganism belonging to the genus Penicillium isolated from the dead body of the larva of Kuwanomeiga was added to the culture medium. By culturing, NO (nitric oxide) and TNF-α
(Tumor necrosis factor) It was found that a novel compound having a production inhibitory activity was produced. The present invention has been completed by further isolating the compound. That is, the present invention has the following general formula:

[0005]

Embedded image (The symbol in the formula, R ¹ represents 2-propyl or 2-butyl,
R ² represents a hydrogen atom or hydroxy. Note that R ¹
Is 2-propyl, R ² represents a hydrogen atom. )
Is a quinolone derivative or a salt thereof, a drug containing the above-mentioned condensed quinolone derivative or a salt thereof as an active ingredient, preferably the drug is a systemic inflammatory response syndrome, sepsis, multiple organ failure, disseminated intravascular coagulation, or It is a drug that is an agent for preventing and treating endotoxin shock.

[0006]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail. The condensed quinolone derivative of the present invention or a salt thereof can be obtained by a conventional method from a culture in which the compound-producing bacterium belonging to the genus Penicillium is cultured in a nutrient medium and the compound is accumulated. As the microorganism used in the method for producing the compound, any microorganism belonging to the genus Penicillium and capable of producing the compound can be used. As such a microorganism, for example, an incomplete filamentous fungus Penicillium sp. EPF-6 strain isolated from the dead larva of Kuwanomeiga larvae can be mentioned. The mycological properties of this strain are as follows.

[0007] 1. Properties in various media (1) Potato and glucose agar media Colonies reach 11 mm in diameter after culturing at 25 ° C for 7 days. The aerial mycelium spreads velvetly on the colony surface, and the center is white and the periphery is olive gray. The back of the colony is light yellow. Conidium formation is very good. (2) Malt extract agar medium Colonies reach a diameter of 15 mm when cultured at 25 ° C. for 7 days. On the colony surface, aerial hyphae swell in the form of wool, giving a light orange color. The back of the colony is light brown. Conidium formation is rather good. (3) Tzapek agar medium The colonies reach a diameter of 21 mm after culturing at 25 ° C. for 7 days. Aerial hyphae spread velvety on the colony surface, giving a bright olive gray color. The back of the colony is dark green. Conidium formation is very good. (4) Sabouraud agar medium After culturing at 25 ° C for 7 days, colonies reach a diameter of 20 mm. On the colony surface, aerial hyphae swell in a wool-like manner, and the center where conidia are formed is bright olive gray and the periphery is brownish white. The back of the colony is light brown. Conidium formation is good. 2. Physiological properties Growth temperature: grows in the range of 10-37 ° C, optimal growth temperature is 20
~ 28 ° C.

[0008] 3. Morphological characteristics The results of observation of the strain grown on the potato / glucose agar medium under an optical microscope are as follows. The hypha has septum, the surface is smooth and branched. The conidiophores do not form a bundle, and most of them are formed upright independently from the basal hypha, but may be formed by branching secondarily. The conidiophore reaches 200 μm in length, 3 to 5.5 μm in diameter, is smooth, colorless and has septum. Approximately three metres are formed at the tip of the conidiophore, and three to five phialides are observed at the tip of the metole. Metre is cylindrical, 10-16μm long and 3-5.5μm wide
It is. Fiaride is oversized, 6-10 μm long and width
2-3 μm. Phyaroconidia are spherical, single cell,
Colorless, smooth surface with a diameter of about 2 μm. No sexual generation is observed. Based on the above mycological properties, the taxonomic position of the EPF-6 strain was determined by The Genera of Funjay Sporrating in Pure Culture 3rd Edition (JAvo
n Arx; The Genera of Fungi Sporulating in Pure C
ulture, 3rd ed. Cramer, Vaduz, published in 1981), this strain was found to be penicillium ( P.
enicillium ). It should be noted that the present strain Penicillium sp. (Penicillium sp.) EPF-6
Deposit No. FERM P-17738 has been deposited at the Institute of Biotechnology and Industrial Technology, Institute of Biotechnology, Tsukuba City, Ibaraki Prefecture, Japan, 1-3-1, Higashi (Postal Code: 305-8566).

(Production method) The compound of the present invention belongs to the genus Penicillium and can be obtained by culturing a microorganism capable of producing the compound of the present invention. The culturing is performed according to a general microorganism culturing method. The medium used for the culture may be any medium as long as the microorganism used can grow, and a synthetic medium, a semi-synthetic medium, or a natural medium can be used. As nutrients to be added to the medium, known nutrients for fungi can be used. For example, commercially available molasses, glucose, maltose, fructose, mannitol, potato starch, corn starch, dextrin, carbohydrates such as soluble starch or fats and oils, fats and the like, as the nitrogen source, nitrogen sources, commercially available peptones ,
Meat extracts, corn steep liquor, cottonseed meal, peanut flour, soy flour, yeast extract, NZ-amine, wheat germ, caseins, fish meal, and inorganic or organic nitrogen sources such as sodium nitrate and ammonium nitrate are used. As the metal salt, lead sulfate such as Na, K, Mg, Ca, Zn, Fe, Mn, Co, and Cu, hydrochloride, nitrate, phosphate, carbonate and the like can be added as required. Valine, leucine,
Amino acids such as isoleucine, phenylalanine, tryptophan, methionine, lysine, arginine, glutamic acid, aspartic acid, and vitamins, oleic acid, methyl oleate, lard oil, silicone oil, secondary metabolite production promoting substances such as surfactants or An antifoaming agent can be used as appropriate. In addition to these, any additives can be used as long as they can be used by the bacteria producing the compound of the present invention and are useful for producing the compound of the present invention.

The culturing method may be the same as that of general antibiotics and the like, and the culturing method may be solid culture or liquid culture. In the case of liquid culture, any of stationary culture, shaking culture, and stirring culture may be performed, but aeration and stirring culture is particularly desirable. As the culture conditions, the culture temperature can be appropriately applied at a temperature at which the producing bacteria can grow and produce the compound of the present invention, that is, within the range of 10 to 37 ° C, but about 28 ° C is preferable. The pH can be appropriately applied in the range of 4 to 9, but is preferably 6 to 7. The cultivation time varies depending on various conditions, usually 1 to 30
The compound of the present invention is accumulated in the medium in about a day. In order to isolate the compound of interest from the culture, the usual extraction and purification means used for isolating metabolites of microorganisms can be appropriately used. The substance in the culture may be a culture solution as it is, or a culture filtrate obtained by centrifuging or adding a filter aid to the culture to obtain a culture filtrate, which is an organic compound that is immiscible with water such as ethyl acetate, chloroform, benzene, and toluene. Add solvent and extract. Further, the culture solution is brought into contact with an appropriate carrier, the produced substance in the filtrate is adsorbed, and then the substance is extracted by eluting with a suitable solvent. For example, Amberlite XAD-2, Diaion HP-20, Diaion CHP-
The substance is adsorbed by contacting with a porous adsorption resin such as 20P or Diaion SP-850. Then methanol,
The substance is eluted using a mixture of water and an organic solvent such as ethanol, acetone or acetonitrile. By increasing the mixing ratio of the organic solvent to a high concentration stepwise or continuously from a low concentration at this time, a fraction having a higher ratio of the substance can be obtained. Next, the substance-containing fraction obtained by using each of the above-mentioned procedures was applied to silica gel, ODS
Further purification and purification can be carried out by a conventional method such as column chromatography using HPLC or the like, centrifugal liquid partition chromatography, high performance liquid chromatography (HPLC) using ODS or the like.

The compounds of the present invention form salts with acids or bases. It is an acid addition salt with an inorganic acid or an organic acid, or a salt with an inorganic base or an organic base, and a pharmaceutically acceptable salt is preferable. Further, the compound of the present invention includes all of various isomers, and also includes hydrates, various solvates and the like. Furthermore, the present invention includes some having polymorphism, and includes all of these crystal forms.

The method of formulating and administering a medicament comprising the compound of the present invention as an active ingredient will be described below in detail. Pharmaceutical compositions containing one or more of the compound of the present invention or a salt thereof as an active ingredient, commonly used carriers and excipients for pharmaceuticals, other additives, tablets, powders, Fine granules, granules,
It is prepared into capsules, pills, solutions, injections, suppositories, ointments, patches, and the like, and is administered orally or parenterally. The clinical dose of the compound of the present invention to humans is appropriately determined in consideration of the patient's symptoms, weight, age, sex, and the like.
Usually 0.1 to 500 mg orally per adult daily, 0.01 to 100 parenteral
mg, which is administered once or in several divided doses.
Since the dose varies under various conditions, a dose smaller than the above dose range may be sufficient. As the solid composition for oral administration according to the present invention, tablets, powders, granules and the like are used. In such solid compositions, the one or more active substances comprise at least one inert diluent, such as lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, starch,
Polyvinylpyrrolidone is mixed with magnesium metasilicate aluminate. The composition may contain, in a conventional manner, additives other than an inert diluent, for example, a lubricant such as magnesium stearate, a disintegrant such as calcium cellulose glycolate, a stabilizer such as lactose, glutamic acid or asparagine. A solubilizing or solubilizing agent such as an acid may be contained. Tablets or pills may be coated with sugar or a film of gastric or enteric substance such as sucrose, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose phthalate, if necessary. Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs and the like, and commonly used inert diluents such as purified water , Ethyl alcohol. The composition may contain, in addition to the inert diluent, solubilizing or solubilizing agents, auxiliaries such as wetting agents and suspending agents, sweetening agents, flavoring agents, fragrances, and preservatives.

[0013] Injections for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Diluents for aqueous solutions and suspensions include, for example, distilled water for injections and physiological saline. Examples of diluents for non-aqueous solutions and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, alcohols such as ethyl alcohol, polysorbate 80 (trade name; polyoxyethylene sorbitan higher fatty acid ester) ). Such a composition is
Further, additives such as a tonicity agent, a preservative, a wetting agent, an emulsifier, a dispersant, a stabilizer (eg, lactose), and a solubilizing or solubilizing agent may be contained. These are sterilized by, for example, filtration through a bacteria retaining filter, blending of a bactericide or irradiation. These can also be used by preparing a sterile solid composition and dissolving in sterile water or a sterile injection solvent before use. When the solubility of the compound of the present invention is low, a solubilization treatment may be performed. As the solubilization treatment, known methods applicable to pharmaceutical preparations, for example, surfactants (polyoxyethylene hydrogenated castor oils, polyoxyethylene sorbitan higher fatty acid esters, polyoxyethylene polyoxypropylene glycols, sucrose fatty acid esters) ), A drug and a solubilizer, for example, a polymer [hydroxypropylmethylcellulose (H
PMC), polyvinylpyrrolidone (PVP), water-soluble polymers such as polyethylene glycol (PEG), carboxymethylethylcellulose (CMEC), hydroxypropylmethylcellulose phthalate (HPMCP), methyl methacrylate-methacrylic acid copolymer Eudragit L , Eudragit S, (trade name); enteric polymer from Rohm and Haas Co., Ltd.]. If necessary, a method for forming a soluble salt, a method for forming an inclusion compound using cyclodextrin or the like can also be employed. The means of solubilization can be appropriately changed depending on the target drug ["Recent formulation technology and its application", Isamu Utsumi, Pharmaceutical Journal 157-159 (1983) and "Pharmaceutical Monograph No. 1, Biology Ability to Use ", Tsuneji Nagai et al., Soft Science, 78-82 (1988))]. Of these, a method of improving the solubility by forming a solid dispersion of the drug and the solubilizing agent is preferably employed (Japanese Patent Application Laid-Open No. 56-493).
No. 14, FR2460667).

[0014]

EXAMPLES The present invention will be described in more detail below, but the present invention is of course not limited to these examples. Example 1 10 g glucose, 20 g potato starch, 5 g polypeptone,
Medium containing yeast extract 5g, calcium carbonate 4g, distilled water 1L
(pH 7.0) was dispensed in 100 ml aliquots into a 500 ml conical flask with a navel and sterilized at 120 ° C. for 20 minutes. Penicillium sp. EP grown well on potato dextrose agar
Scrape and inoculate the F-6 strain and incubate at 28 ° C and 200 rpm.
The cells were cultured with shaking for 4 days. The above culture solution was inoculated at a rate of 3% into four culture media prepared in the same manner, and incubated at 28 ° C. and 200 rpm.
After shaking culture for one day, a seed culture solution was obtained. Next, as a production medium, 30 g of brown rice, 60 mg of yeast extract, 30 mg of monosodium phosphate, 50 ml of distilled water were placed in a 500 ml Erlenmeyer flask.
Was prepared and sterilized twice at 120 ° C. for 20 minutes. This medium is inoculated with a seed culture solution at a rate of 3%, and incubated at 28 ° C for 21
The stationary culture was performed for one day.

20 L of acetone was added to the 5 L of the culture thus cultured, and the mixture was stirred for 3 hours. Then 5% radio light
After adding # 600 (Showa Kagaku), the mixture was filtered, and acetone was distilled off from the filtrate under reduced pressure. This concentrated solution (pH 7) was adsorbed on 300 ml of Diaion HP-20 (Mitsubishi Chemical), and this was washed with 1 L of water. Next, after flowing 1.2 L of 20% acetone water, 40% acetone water
Eluted at 1.2L. Acetone was distilled off under reduced pressure from the fraction eluted with 40% acetone. The concentrated solution was partitioned and extracted with an equal volume of chloroform in two layers. The obtained chloroform layer was concentrated to dryness under reduced pressure to obtain 1.53 g of an extract. The obtained extract was subjected to column chromatography using 30 g of silica gel C300 (Wako Pure Chemical Industries, Ltd.). Silica gel column chromatography
Was eluted with a mixed solvent of chloroform-methanol to gradually increase the content of methanol. First, 150 ml of the mixed solvent (100: 1) is flowed, and sequentially (50: 1), (20: 1),
(10: 1) and (5: 1) were eluted by flowing 120 ml each. Chloroform-
The desired activity was observed in the fraction eluted with methanol (5: 1), and the obtained fraction was concentrated to dryness under reduced pressure to obtain 970 mg of a brown substance. The obtained active fraction was subjected to column chromatography of Sephadex LH-20 (Pharmacia) 3 cm × 90 cm. For elution, methanol was used to fractionate 5 ml each. The obtained active fractions and fractions 42 to 47 were concentrated to dryness under reduced pressure to obtain 390 mg of an active substance. The obtained active substance was finally fractionated using high performance liquid chromatography. High-performance liquid chromatography conditions are Supelc
osil ABZ + plus (21.2mm id × 250mm, manufactured by Spelco)
Was used as a solvent, and 30% acetonitrile was flowed at a flow rate of 5 ml / min. Detection was performed at UV 275 nm, and activity was observed in the peak eluted at 20 minutes. The peak was collected and concentrated under reduced pressure to isolate 81 mg of Compound A. 15 minutes and 22
The activity eluted in the peak eluted per minute was observed. From the respective peaks, 21 mg of Compound B and 21 mg of Compound C were obtained in the same manner.
8 mg was isolated.

Column: Supelcosil ABZ + plus (21.2 mm id × 250 mm, manufactured by Spelco) Elution solvent: mixed solution of acetonitrile-water (30:70) Outflow rate: 5 ml / min Detection wavelength: 275 nm Has chemical properties. (1) Color and shape: white amorphous translucent solid. (2) Classification of acidic, neutral and basic: neutral. Solubility: soluble in methanol and chloroform, but water,
Hardly soluble in hexane. (4) UV absorption spectrum: 215, 248, 256, 315, 32
The absorption maximum is shown at 7 nm. (5) Molecular weight: 270 (6) Molecular formula: C ₁₆ H ₁₈ N ₂ O ₂ (7) Infrared absorption spectrum (KBr): (FIG. 1) (8) ¹ H-NMR spectrum (400 MHz, DMSO-d ₆ ) : (FIG. 2) (9) ¹³ C-NMR spectrum (100 MHz, DMSO-d ₆ ): (FIG. 3) From the above physicochemical properties, the chemical structural formula of compound A was determined as follows.

[0017]

Embedded image

Compound B has the following physicochemical properties. (1) Color and shape: white amorphous translucent solid. (2) Classification of acidic, neutral and basic: neutral. Solubility: soluble in methanol and chloroform, but water,
Almost insoluble in hexane (4) Ultraviolet absorption spectrum: 214, 248, 255, 314, 32
The absorption maximum is shown at 7 nm. (5) Molecular weight: 256 (6) Molecular formula: C ₁₅ H ₁₆ N ₂ O ₂ (7) Infrared absorption spectrum (KBr): (FIG. 4) (8) ¹ H-NMR spectrum (400 MHz, DMSO-d ₆ ) : (FIG. 5) (9) ¹³ C-NMR spectrum (100 MHz, DMSO-d ₆ ): (FIG. 6) From the above physicochemical properties, the chemical structural formula of compound B was determined as follows.

[0019]

Embedded image Compound C has the following physicochemical properties. (1) Color and shape: white amorphous translucent solid. (2) Classification of acidic, neutral and basic: neutral. Solubility: soluble in methanol and chloroform, but water,
Almost insoluble in hexane (4) Ultraviolet absorption spectrum: 215, 250, 258, 315, 32
The absorption maximum is shown at 8 nm. (5) Molecular weight: 286 (6) Molecular formula: C ₁₆ H ₁₈ N ₂ O ₃ (7) Infrared absorption spectrum (KBr): (FIG. 7) (8) ¹ H-NMR spectrum (400 MHz, DMSO-d ₆ ) : (FIG. 8) (9) ¹³ C-NMR spectrum (100 MHz, DMSO-d ₆ ): (FIG. 9) From the above physicochemical properties, the chemical structural formula of compound C was determined as follows.

[0020]

Embedded image

[0021]

The compound of the present invention is useful as a prophylactic and therapeutic agent for systemic inflammatory response syndrome, sepsis, multiple organ failure, disseminated intravascular coagulation syndrome, endotoxin shock, etc. based on the inhibitory action of NO and TNF-α production. is there.

The NO production inhibitory activity test and the TNF-α production inhibitory activity test are described below. NO production inhibition activity test J774.1 cells (BALB / c blood-derived macrophage cell line,
RPMI16 containing 10% FBS (fetal bovine serum) with National Institute of Health and Medical Care Research Institute Infection and Tropical Diseases Research Division Switch Teruo et al.
The cells were cultured in 40 media (37 ° C., 5% CO ₂ ), and when the cells became floating, the culture medium was replaced and the cells were cultured overnight. The culture was transferred to a centrifuge tube and centrifuged at 1,000 × g for 10 minutes. Remove the supernatant and resuspend the cells in RPMI 1640 medium containing 2% FBS,
It was diluted with the same medium to 1.25 × 10 ⁶ / ml. The cell solution was seeded at 160 μl / well on a 96-well flat bottom microplate (Nippon Becton Dickinson Co., Ltd.). The test compound was dissolved in DMSO, adjusted to 100 mg / ml, and used as a stock solution.
400, 800, 160 in RPMI1640 medium containing 2% FBS
It was diluted 0, 3200 times. These diluents were prepared previously96
20 μl / well was added to the well microplate. Next, IFN-γ
20 U / ml at 10 μl / well, Salmonella minnesota LPS
(Sigma Chemical Co.) 200 ng / ml was added at 10 μl / well. After culturing for 48 hours, the culture supernatant and the nitrite standard solution are mixed.
100 μl was placed in a 96-well microplate. Equivalent amount of grease reagent (1: 1, v / v, N- [1-naphthyl] ethyl) in all wells
enediamine dihydrocholride (NED, Sigma Chemical C
o.), 0.1% in H ₂ O, sulfanilamide 1.0% in 5% H ₂ PO ₄ )
Was added and the color was developed. Biomek 1000 spectrophotometer
(Beckman Instruments, Palo Alto, Calif.) The absorbance was measured at 540 nm to determine the NO production inhibition rate. For example, Compound A suppressed LPS-induced NO production in J774.1 cells.

TNF-α production inhibitory activity J774.1 cells (macrophage cell line derived from BALB / c blood,
RPMI containing 10% of FBS (fetal bovine serum) from Infectious Diseases and Tropical Diseases Research Division
The cells were cultured in a 1640 medium (37 ° C., 5% CO ₂ ), and when the cells became suspended, the culture medium was replaced and the cells were cultured overnight. The culture was transferred to a centrifuge tube and centrifuged at 1,000 × g for 10 min. The supernatant was removed, the cells were resuspended in RPMI1640 medium containing 2% FBS, and diluted with the same medium so that the number of cells became 1.25 × 10 ⁶ / ml.
The cell solution was seeded at 160 μl / well on a 96-well flat bottom microplate (Nippon Becton Dickinson Co., Ltd.).
The test compound was dissolved in DMSO, adjusted to 100 mg / ml, and used as a stock solution. This stock solution is 100, in RPMI1640 medium containing 5% FBS.
It was diluted 200, 400, 800, 1600 times. These diluted solutions were added to the previously prepared 96-well microplate at 20 μl / well. Then RPMI1640 medium containing 2% FBS or Salmonell
a minnesota LPS (Sigma Chemical Co.) 100 ng / ml for 20
μl / well was added. After culture for 3.5 hours, culture supernatant 160 μl
Was transferred to a 96-well microplate and stored frozen (-60 ° C).

L929 (mouse fibroblasts, distributed by Teruo Switching, Division of Infection and Tropical Diseases, National Institute of International Health) was cultured in RPMI1640 medium containing 5% FBS. After the cells proliferated, the culture solution was removed, the cells were washed with PBS, trypsin was added, and the cells were detached from the surface of the flask. FBS 5
% RPMI1640 medium was added to the flask, transferred to a centrifuge tube, and centrifuged at 1,000 g for 10 min. Remove supernatant and transfer cells to FB
Resuspend in RPMI1640 medium containing 5% S, and count cells 5 × 10 ⁵
A cell solution diluted with the same medium to a concentration of / ml was seeded on a 96-well microplate at 100 µl / well. After culturing for 20 hours, 5μ
After culturing for 3 hours in RPMI1640 medium (containing 5% FBS) containing actinomycin D (Sigma Chemical Co.) at a concentration of g / ml, TNF-α
The standard solution and the cryopreserved sample were serially diluted (2 to 2048 times) on the culture plate. After culturing for 18 hours, the culture solution is discarded, the cells are washed with PBS, and tetrazolium salt (MTT, Sigma Chl.
emical Co.) was added at 100 μl / well with RPMI1640 medium (without adding phenol red) containing 1 mg / ml. After 3 hours of culture,
The supernatant was removed and propanol was added to dissolve the product.
Biomek 1000 spectrophotometer (Beckman Instrument
s, Palo Alto, Calif.) The absorbance was measured at 540 nm, the number of living cells was compared, TNF-α in the sample was quantified, and the inhibition rate was determined. For example, Compound A suppressed TNF-α production of J774.1 cells by LPS.

[Brief description of the drawings]

FIG. 1 shows an infrared absorption spectrum (KBr) of Compound A.

FIG. 2 ¹ H-NMR spectrum of compound A (400 MHz, DMSO-
d ₆ ).

FIG. 3 ¹³ C-NMR spectrum of compound A (100 MHz, DMSO-d
₆ ) is shown.

FIG. 4 shows an infrared absorption spectrum (KBr) of Compound B.

FIG. 5 ¹ H-NMR spectrum of compound B (400 MHz, DMSO-
d ₆ ).

FIG. 6 shows a ¹³ C-NMR spectrum of compound B (100 MHz, DMSO-d
₆ ) is shown.

FIG. 7 shows an infrared absorption spectrum (KBr) of Compound C.

FIG. 8 shows a ¹ H-NMR spectrum (400 MHz, DMSO-
d ₆ ).

FIG. 9 shows a ¹³ C-NMR spectrum of compound C (100 MHz, DMSO-d
₆ ) is shown.

Continued on the front page (51) Int.Cl. ⁷ Identification FI FI Theme Court II (Reference) A61P 31/04 A61P 31/04 (72) Inventor Noriharu Takahashi 1-2 Toyodaidai, Utsunomiya City, Tochigi Prefecture Teikyo University Dormitory C- 402 (72) Inventor Koji Nagai 1-1-8 Shozusawa, Itabashi-ku, Tokyo Yamanouchi Pharmaceutical Co., Ltd. (72) Inventor Koichi Tanaka 1-1-8 Shozusawa, Itabashi-ku, Tokyo Yamanouchi Pharmaceutical Co., Ltd. (72) Inventor Yoji Yamaguchi 1-1-8 Shozuzawa, Itabashi-ku, Tokyo Yamanouchi Pharmaceutical Co., Ltd. (72) Inventor Tatsuhiro Tokunaga 21 Miyukigaoka, Tsukuba-shi, Ibaraki Yamanouchi Pharmaceutical Co., Ltd. JJ04 KK02 LL04 PP01 4C086 AA01 AA02 AA03 CB05 MA01 MA04 NA14 ZA54 ZB11 ZB35

Claims (3)

[Claims] (1) The following formula: (The symbol in the formula, R ¹ represents 2-propyl or 2-butyl,
R ² represents a hydrogen atom or hydroxy. Note that R ¹
Is 2-propyl, R ² represents a hydrogen atom. )
Or a salt thereof. 2. A medicament comprising the quinolone derivative according to claim 1 or a salt thereof as an active ingredient. 3. The medicament according to claim 2, which is an agent for preventing and treating systemic inflammatory response syndrome, sepsis, multiple organ failure, disseminated intravascular coagulation syndrome, or endotoxin shock.