JP3273918B2 - Anti-Helicobacter pylori agent - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to pharmaceuticals, particularly to an anti-Helicobacter pylori agent useful for the treatment and prevention of various diseases caused by Helicobacter pylori infection.

[0002]

2. Description of the Related Art Helicobacter pylori (Helic
obstructor pylori is a pathogenic bacterium discovered in 1983, and has peptic ulcer (eg, gastric ulcer or duodenal ulcer), inflammation (eg, gastritis), diseases of the upper gastrointestinal tract such as gastric cancer, MALT (mucosa-mucosa). associated
lymphoid tissue) It is said to be a lymphoid species or a background pathogen of chronic heart disease. At present, studies on the treatment of Helicobacter pylori infection are being actively conducted, and many such treatments have been reported as follows, including those for the purpose of eradication and those for the prevention of recurrence. For example, bismuth, antibiotics, proton pump inhibitors (PP
I) or a single-drug administration such as an anti-ulcer drug or a multi-drug combination method (drug combination, triple-drug combination) combining the aforementioned drugs and the like (Internal Medicine, Special Issue, Vol. 78, No. 1 (1996), Nankodo).
However, the above treatments may require, for example, frequent dosing, or require large doses over normal doses.
There are many points that need to be resolved, such as the onset of diarrhea and constipation due to drug administration, and the development of resistant bacteria. Further, a substance useful as an anti-Helicobacter pylori agent related to the present invention is disclosed in JP-A-7-189 as YL-02729S substance, and its use is particularly antibacterial against methicillin-resistant Staphylococcus aureus. The patent application includes YL
There is no suggestion or report on the antibacterial activity of the -02729S substance against Helicobacter pylori.

[0003]

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[0004]

SUMMARY OF THE INVENTION An object of the present invention is to provide an excellent anti-Helicobacter pylori agent.

[0005]

Means for Solving the Problems The present inventors have proposed the above YL.
As a result of intensive studies on the substance -02729S, it was found that the substance has excellent anti-Helicobacter pylori activity, and that the substance has high selectivity and does not affect other bacteria. Hereinafter, the present invention will be described in detail. The present invention provides: 1) the following formula (I):

[0006]

Embedded image (The symbol R1 in the formula represents a hydrogen atom or a hydroxyl group.) 2- (2-Nonenyl) -3-methyl-
An anti-Helicobacter pylori agent comprising a 4 (1H) -quinolone derivative or a pharmaceutically acceptable salt thereof as an active ingredient, preferably R1 is a hydroxyl group, more preferably an anti-Helicobacter pylori agent according to (1). Is 2) the following general formula (II)

[0007]

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(The symbol R1 in the formula represents a hydrogen atom or a hydroxyl group.) 2- (2-trans-nonenyl) -3-methyl-4 (1H) -quinolone derivative or a pharmaceutically acceptable derivative thereof An anti-Helicobacter pylori agent containing an acceptable salt as an active ingredient, and most preferably the anti-Helicobacter pylori agent according to 2), wherein R1 is a hydroxyl group. The invention also relates to the use of a composition comprising a substance of the formula (I) for the prevention or treatment of Helicobacter pylori infection, and to the use of a substance of the formula (I) and other agents, preferably antibiotics, acids. Also includes simultaneous or timed administration with the drug, H2 blocker, and use to prevent recurrence of disease caused by Helicobacter pylori infection. Further, a prodrug form of the substance of the formula (I) obtained by a conventional method is also included in the present invention. Since the active ingredient (I) of the anti-Helicobacter pylori agent of the present invention has a double bond, it has a cis form, tra
There are ns forms and tautomers. The existence of tautomers is disclosed in JP-A-7-189 as follows.

[0009]

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The pharmaceutically acceptable salt is a salt with an acid addition salt with an inorganic acid or an organic acid, and a pharmaceutically acceptable salt is preferred. Specific examples of these salts include mineral acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, and phosphoric acid, or formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, and succinic acid. And organic acids such as fumaric acid, maleic acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid and ethanesulfonic acid, and acid addition salts with acidic amino acids such as aspartic acid and glutamic acid. Further, hydrates or various solvates of the active ingredient of the present invention are included. Further, the active ingredient may have a polymorphism, and includes all of those crystal forms.

The typical production of the active ingredient of the anti-Helicobacter pylori agent of the present invention is exemplified by the production method disclosed in JP-A-7-189. A method for obtaining the active ingredient by a fermentation method is a method for producing a bacterium belonging to the genus Arthrobacter, for example, Arthrobacter sp. Of Deposit No. FERM BP-6326, which is internationally deposited at the Institute of Biotechnology and Industrial Technology. .) YL-0272
It is preferable to use a 9S strain, culture the strain, and isolate and purify from the culture according to a conventional method.

The formulation and administration of the anti-Helicobacter pylori agent of the present invention will be described below in detail. Pharmaceutical compositions containing one or more of the substance represented by formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient include commonly used carriers and excipients for pharmaceutical preparations, Tablets, powders, fine granules, granules, capsules, pills,
It is prepared into solutions, injections, suppositories, ointments, patches, etc., and is administered orally or parenterally. The clinical dose of the present invention for humans is appropriately determined in consideration of the symptoms, weight, age, sex, etc. of the patient to which the present invention is applied.

Normally 0.1 to 500 m orally per day for an adult
g, parenterally, 0.01 to 100 mg, administered once or in several divided doses. Since the dosage varies under various conditions, an amount smaller than the above dosage range may be sufficient. The anti-Helicobacter pylori agent of the present invention can be used in combination with another drug, for example, an antibacterial agent to be described later, or simultaneously or at a later time. As the solid composition for oral administration according to the present invention, tablets, powders, granules and the like are used. In such a solid composition, one or more of the active substances
It is mixed with at least one inert diluent, such as lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, starch, polyvinylpyrrolidone, metasilicate, magnesium 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 agent such as an acid or a solubilizing agent may be contained. Tablets or pills may be coated with a film of a 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, For example, it contains purified water and ethyl alcohol. The composition may contain, besides an inert diluent, solubilizing or solubilizing agents, auxiliaries such as wetting agents and suspending agents, sweetening agents, flavoring agents, flavoring agents,
Preservatives may be included. Injections for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions,
Emulsifying agents. 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, and polysorbate 80 (trade name: polyoxyethylene sorbitan higher fatty acid ester). ). Such compositions may further comprise a tonicity agent, a preservative, a wetting agent,
Additives such as emulsifiers, dispersants, stabilizers (eg, lactose), solubilizing or solubilizing agents may be included. These are sterilized by, for example, filtration through a bacteria retaining filter, blending of a bactericide or irradiation. They 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, it may be subjected to a solubilization treatment. As a 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 such as a polymer (hydroxypropylmethylcellulose (HPM
C), water-soluble polymers such as polyvinylpyrrolidone (PVP) and polyethylene glycol (PEG), carboxymethylethylcellulose (CMEC), hydroxypropylmethylcellulose phthalate (HPMCP), methyl methacrylate-methacrylic acid copolymer (Eudragit L) , S, trade name; manufactured by Rohm and Haas)
Etc.) to form a solid dispersion. If necessary, a method for converting to a soluble salt,
For example, cyclodextrins or dextrins such as α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, hydroxyalkylated cyclodextrin such as hydroxypropyl β cyclodextrin, methylated cyclodextrin, and dimethyl β cyclodextrin are used. For example, a method of forming an inclusion compound by the method. The means of solubilization can be appropriately changed according to the target drug ["Recent formulation technology and its application", Isamu Utsumi, Pharmaceutical Journal 157-1
59 (1983) and “Pharmaceutical Monograph No. 1, Bioavailability”, Tsuneji Nagai et al., Soft Science, 78
-82 (1988)]. Among them, a method of improving the solubility by forming a solid dispersion of the drug and the solubilizer is preferably used (Japanese Patent Application Laid-Open No. 56-49314, FR24).
No. 60667).

According to the present invention, the active compound is used not only alone but also in combination with other antibacterial agents and the like (preferably 1
~ 3 types) can be used. As described above, the dosage form can be used simultaneously with the anti-Helicobacter pylori agent of the present invention or at a later time. Such other antibacterial agents include, for example, nitroimidazole antibiotics (eg, tinidazole and metronidazole), tetracyclines (eg, tetracycline, minocycline, or doxycycline), penicillins (eg, amoxicillin, ampicillin, tarampicillin, bacampicillin, Lenampicillin, mezlocillin,
Sultamicillin), cephalosporins (eg, cefaclor, cefadroxil, cephalexin, cefpodoxime proxetil, cefixime, cefdinir, ceftibutene, cefotiamhexetil, cefmetmet pivoxil, or cefuroxime axetil), penem Drugs (eg, flopenem or lipipenem acoxil), macrolide drugs (eg, erythromycin, oleandomycin, josamycin, midecamycin, rokitamicin, clarithromycin, roxithromycin, or azithromycin), lincomycins Drugs (eg, lincomycin or clindamycin), aminoglycoside drugs (eg, paromomycin), quinolone drugs (eg, ofloxacin, levofloki) Singh, it norfloxacin, enoxacin, ciprofloxacin, lomefloxacin, tosufloxacin, fleroxacin, sparfloxacin, temafloxacin, Najifokisashin, be mentioned grepafloxacin, or Pazufokisashin etc.) and nitrofurantoin. In addition, pharmaceutical compounds used for the treatment of acid-related diseases {eg, acid pump inhibitors (eg, omeprazole or lansoprazole)), H2 antagonists (eg, ranitidine, cimetidine, or famotidine), or inhibition of Helicobacter pylori gastric mucosal adhesion. Combinations of an active gastric mucosal protective agent and the active compounds are also included within the scope of the present invention.

[0017]

BEST MODE FOR CARRYING OUT THE INVENTION

The present invention will be described in more detail below. Of course, it is not limited to these examples.

Example 1 An in vitro test of the substance of the present invention was performed by the following method. Measurement of antibacterial activity (1) Preparation of agar plate containing antibacterial substance The substance to be evaluated was 100% dimethyl sulfoxide (DMS).
O) and serially diluted 2-fold. This diluted solution was put into a sterilized round petri dish, and sterilized there and kept at 50 ° C.
0 ml of Brucella agar medium (0.1% β-cyclodextrin) was added, mixed, and hardened. The final concentration of DMSO will be less than 1%. (2) Preparation of inoculum and determination of results Using a Brucella agar medium (containing 5% calf serum), a multi-gas incubator (N ₂ : 80%, CO ₂ : 15%,
(O ₂ : 5%) at 37 ° C. for 3 days, for example, Helicobacter pylori ATCC
The 43504 was prepared using a Brucella broth to be about 10 ^8/1 ml by turbidity. A solution obtained by diluting this bacterial solution 100-fold with brucella broth in the same manner was added to a drug-containing agar medium for about 5 minutes using a microplanter.
μl was inoculated on the agar surface. The inoculated agar plate is cultured in the above-mentioned multigas incubator at 37 ° C. for 3 days (72 hours). The agar plate after completion of the culture was observed, and the minimum drug concentration at which no growth was observed was defined as the MIC. As a result, the MIC of the anti-Helicobacter pylori agent of the present invention was 0.1.
It was 025 μg / ml.

Example 2 An in vitro test of the substance was carried out by the following method. Measurement of antibacterial activity Preparation of agar plate containing antibacterial substance 100% dimethyl sulfoxide (DMS)
O) and serially diluted 2-fold. This diluted solution was put into a sterilized round petri dish, and sterilized there and kept at 50 ° C.
The mixture was added to 0 ml of Brucella agar medium (0.1% β-cyclodextrin), mixed and solidified. The final concentration of DMSO will be less than 1%. Preparation of Inoculum and Judgment of Results Using a brucella agar medium (containing 5% calf serum), a multi-gas incubator (N ₂ : 80%, CO ₂ : 15%,
(O ₂ : 5%) at 37 ° C. for 3 days, for example, Helicobacter pylori ATCC
The 43504 was prepared using a Brucella broth to be about 10 ^8/1 ml by turbidity. A solution obtained by diluting this bacterial solution 100-fold with brucella broth in the same manner was added to a drug-containing agar medium for about 5 minutes using a microplanter.
μl was inoculated on the agar surface. The inoculated agar plate is cultured in the above-mentioned multigas incubator at 37 ° C. for 3 days (72 hours). The agar plate after completion of the culture was observed, and the minimum drug concentration at which no growth was observed was defined as the MIC. As a result, 1-hydroxy-2- (2-trans-nonenyl)-
The MIC of 3-methyl-4 (1H) -quinolone is 0.02
It was 5 μg / ml.

Example 3 An in vitro test for facultative anaerobic bacteria and aerobic bacteria was carried out by the following method. Preparation of agar plate containing antibacterial substance The substance to be evaluated was 100% dimethyl sulfoxide (DMS).
O) and serially diluted 2-fold. This diluted solution was put into a sterilized round petri dish, and sterilized there and kept at 50 ° C.
Add to 0 ml of Mueller Hinton agar medium and mix.
Hardened. The final concentration of DMSO will be less than 1%. Preparation of Inoculum and Judgment of Result Using a Mueller Hinton broth, a bacterial solution cultured overnight in a Flan apparatus set at 37 ° C. was diluted with Mueller Hinton broth to about 10 ⁶ cells / ml. About 5 μl of this bacterial solution was inoculated on the agar medium containing the drug using a microplanter. The inoculated agar plate was cultured for 18 hours in a 37 ° C. furan apparatus. The agar plate after completion of the culture was observed, and the minimum drug concentration at which no growth was observed was defined as the MIC.

Result 1-hydroxy-2- (2-trans-nonenyl) -3-
Methyl-4 (1H) -quinolone is available from Staphylococcus aureus.
us) FDA209P, Escherichia coli (Esch)
erichia coli) O-1, Pseudomonas
Elginosa (Pseudomonasaerugino)
sa) MIC against representative facultative anaerobic bacteria and aerobic bacteria such as NCTC10490 showed a value of 12.5 μg / ml or more.

Example 4 An in vitro test for anaerobic bacteria was performed by the following method. Preparation of agar plate containing antibacterial substance The substance to be evaluated was 100% dimethyl sulfoxide (DMS).
O) and serially diluted 2-fold. This diluted solution was put into a sterilized round petri dish, and sterilized there and kept at 50 ° C.
The mixture was added to 0 ml of GAM agar medium, mixed and solidified. DM
The final concentration of SO will be less than 1%. Preparation of inoculum and determination of results Using GAM broth, N ₂ 80%; CO ₂ 10%; H
_2. A bacterial solution cultured overnight at 37 ° C. using an anaerobic bacterium culture apparatus gas-replaced with a 10% mixed gas was prepared using the same GAM broth to a concentration of about 10 ⁶ cells / ml. About 5 μl of this bacterial solution was inoculated on the agar medium containing the drug using a microplanter. The inoculated agar plate was cultured for 18 hours in the anaerobic bacterium incubator set at 37 ° C. as described above. The agar plate after completion of the culture was observed, and the minimum drug concentration at which no growth was observed was defined as the MIC.

Result 1-hydroxy-2- (2-trans-nonenyl) -3-
Methyl-4 (1H) -quinolone is available from Bifidobacterium bifidum (Bifidobacterium b.
ifidum) CAYA21-1, Peptostreptococcus products (Peptostreptocos)
cactus product CAYA12-2, Bacteroides fragilis (Bacteroides)
MICs for obligate anaerobic bacteria, such as G. fragilis) GAI5562, showed values greater than 25 μg / ml.

Example 5 In vivo antibacterial activity against Helicobacter pylori was measured by the following method. In vivo antimicrobial activity Infection experiments were performed using gerbils that have been reported to have stable infection (J. Gastroenterology 31: supple
IX, 24-28, 1996). Mongolian fasting overnight (Mongo
5% for lian gerbils, (MGS / Sea), male, 4-week-old)
Heli cultured overnight in Brucella broth supplemented with calf serum
A bacterial solution of cobacter pylori ATCC43504 was inoculated into the stomach using a sonde. Start treatment about 1 week after infection,
The drug to be evaluated is dissolved in a solvent according to a standard method, and 10 mg / kg, 1
The dose of mg / kg or 0.1 ml / kg was orally administered twice a day for 3 days. The day after the end of administration, the stomach was removed and crushed. The stomach homogenate solution was serially diluted 10-fold, inoculated in a modified skilow medium, and incubated at 37 ° C., 6 to 6 under microaerobic conditions or 10% CO ₂ conditions.
Cultured for 7 days. The number of bacteria in the stomach was calculated from the number of bacteria that had grown. As a result, 1-hydroxy-2- (2-trans-
Nonenyl) -3-methyl-4 (1H) -quinolone was confirmed to reduce the number of bacteria in the stomach.

[0025]

Industrial Applicability The present invention has an antibacterial activity against Helicobacter pylori and is effective for treating Helicobacter pylori in humans and related Helicobacter genus infections in animals. In addition, the anti-Helicobacter pylori agent of the present invention can be used for peptic ulcers (eg, gastric and duodenal ulcers), inflammation (acute or chronic gastritis or duodenitis), diseases of the upper gastrointestinal tract such as gastric cancer, MALT (muco).
sa-associated lymphoid tissue) It is effective for prevention (including prevention of recurrence) and treatment of lymphatic species or chronic heart disease.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI C07D 215/60 C07D 215/60 (72) Inventor Koichi Tanaka 1-1-8 Shozuzawa, Itabashi-ku, Tokyo Yamanouchi Pharmaceutical Co., Ltd. (72 ) Inventor Masayuki Komiya 1-1-8 Shozuzawa, Itabashi-ku, Tokyo Yamanouchi Pharmaceutical Co., Ltd. (72) Inventor Ratna Murni Lantia Tomojo Indonesia Jakarta 11510 Art R 008 RV 008 RV 009 Complex Greenville Block Quo Number 12 (56 References: JP-A-7-189 (JP, A) WO 97/12868 (WO, A1) Journal of Antibiotics, (1996), Vol. 49, No. 8, p. 823-82 (58) Field surveyed (Int. Cl. ⁷ , DB name) A61K 31/47 C07D 215/22 C07D 215/60 CA (STN) REGISTRY (STN)

Claims (1)

(57) [Claims] 1. A compound represented by the following general formula (II) : (The symbol R1 in the formula represents a hydroxyl group.) 2- (2- trans- nonenyl) -3-methyl-4
An anti-Helicobacter pylori agent comprising a (1H) -quinolone derivative or a pharmaceutically acceptable salt thereof as an active ingredient.