JP2012246228A - Antimicrobial agent against legionella - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an antimicrobial agent targeting Legionella.SOLUTION: The antimicrobial agent is based on unexpected findings that anthranilic acid or its derivatives, or ribose thereof or deoxyribulose derivatives can kill Legionella. The obtained antimicrobial agent using a compound having a relatively simple structure can kill Legionella. The obtained antibacterial substance is confirmed to have specific bactericidal property on Legionella. Namely, the bactericidal substance is extremely useful as a safe bactericide for selectively killing Legionella.

Description

  The present invention relates to the field of antimicrobial agents. More specifically, the present invention relates to Legionella antibacterial agents.

  Legionella is a general term for eubacteria belonging to the genus Legionella, and is a Gram-negative bacilli. Includes species that cause many Legionellosis such as Legionella pneumonia (Military illness). Legionella are bacteria that are normally present in the environment and are usually less likely to cause infections. However, in an environment that is susceptible to infection, especially for those with low resistance, such as the elderly, Legionella pneumophila is mainly responsible for Legionella infections (Legionella pneumonia and Pontiac fever) in humans. Become.

  Legionella bacteria can also cause slime damage such as slime adhering to the inside of the piping due to intrusion into the cooling water system and blocking the piping or reducing the heat exchange rate. As a technical guideline notification regarding measures necessary to prevent legionellosis (Ministry of Health, Labor and Welfare Notification No. 264), this situation is prevented by replacing cooling water, disinfecting, cleaning cooling equipment, etc. It is supposed to be possible. However, as for disinfection, only chlorinated chemicals are illustrated, and in this case, a great deal of labor is spent on maintaining and maintaining free chlorine concentration. Is done.

  And since it is an intracellular parasitic bacterium, the efficacy of a drug with poor penetration into cells is weak. Also, since many produce β-lactamase, penicillin and cephem antibacterial drugs are not effective. It has been conventionally practiced to administer macrolide, new quinolone, rifampicin, and tetracycline antibacterial agents that are highly migrating into phagocytes.

Anthranilic acid (AA) and its analogs are found in opportunistic bacteria Pseudomonas.
An antibacterial effect against aeruginosa (Pa) has been suggested (Non-patent Document 1). However, there is no description about the antibacterial activity against Legionella, and it can be said that the effect on Legionella can not be predicted from the effect on Pa and the effect on Pa. Moreover, as far as the present inventor is aware, there is no report referring to the antibacterial activity of anthranilic acid against Legionella.

  Non-Patent Document 2 describes that the synthesis of autoinducer (AI) synthesized from anthranilic acid (AA) is inhibited by AA analog. However, there is no description about antibacterial activity against Legionella, and no suggestion has been made.

  Non-Patent Document 3 is a paper on the antibacterial properties of plant hormone derivative indole 3 propionic acid against Legionella pneumophila (Lp). Here, it is only described that tryptophan has antibacterial properties.

  Non-patent document 4 describes the killing of Legionella, but only describes the anti-legionella peptide produced by Staphylococcus warneri.

  Patent Document 1 describes a Legionella-producing antibacterial substance. However, no specific substances are disclosed in this document. However, the disclosed content is merely an assembly of hydrophobic small molecules, and only a combined effect is described.

Patent Document 2 relates to a transparent medium for culturing Legionella, but there is no description regarding killing of Legionella.

JP 2000-053514 A Japanese Patent No. 4021969

Lesic B. , Et al. , PloS Pathog 2007, 3 (9): e126, 1229-1239. Calfee MW, et al. , PNAS 2001, 98: 11633-11637. Mandelbaum-Shavit F et al. , Antimicrobial Agents and Chemotherapy 1991, 35 (12) 2526-2530. Hechard Y et al. , FEMS Microbiology Letters 252 (2005), 19-23.

  This invention makes it a subject to provide the novel chemical | medical agent which can kill Legionella bacteria.

  The above problems have been solved by unexpectedly finding that anthranilic acid or a derivative thereof has the ability to kill Legionella.

  Accordingly, the present invention provides the following.

  In one aspect, the present invention provides the following formula:

and

And at least a compound selected from the group consisting of ribosyl derivatives and deoxyribulose derivatives thereof, or a phospho form, a salt or a solvate thereof,
An antibacterial agent against Legionella is provided.

  In one embodiment, the compound is included at a concentration of at least 0.01 mM.

  In another embodiment, the compound is contained in a concentration from 0.1 mM to 10 mM.

  In another embodiment, the compound is contained in a concentration from 1.0 mM to 10 mM.

  In another embodiment, the compound is

,

It is.

  The fluoro derivative can be any of the following:

  In another embodiment, the antimicrobial agent of the present invention comprises an additional antimicrobial substance.

  In another embodiment, the antimicrobial agent of the present invention is solid or liquid.

  In another aspect, the present invention provides a pharmaceutical composition comprising the antibacterial agent of the present invention.

  In another aspect, the present invention provides an article comprising the antibacterial agent of the present invention.

  In one embodiment, the antimicrobial agent is coated on the article.

  In another embodiment, the article is selected from a container, bath tub, handrail, toothbrush, filter, mask, tile, shower head, bath tub, deck brush, hose and nebulizer.

  In another embodiment, the antibacterial agent includes at least one selected from the group consisting of an inorganic coating agent, an organic coating agent, and a wax as a base.

  In another aspect, the present invention provides a method for inhibiting or killing Legionella, wherein the antibacterial agent according to any one of claims 1 to 7 is contacted with Legionella in an effective amount for inhibiting or killing. A method comprising the steps is provided.

  In one embodiment, the contacting is performed in the presence of less than 0.7M sodium chloride (NaCl).

  Preferred embodiments of the present invention will be shown below, but those skilled in the art can appropriately implement the embodiments and the like from the description of the present invention and well-known and common techniques in the field, and the effects and effects of the present invention can be easily achieved. It should be recognized to understand.

  The present invention can kill Legionella using a compound having a relatively simple structure. Moreover, it was confirmed that the bactericidal substance of the present invention has a bactericidal property specific to Legionella. That is, the bactericidal substance of the present invention is extremely useful as a safe bactericidal agent for selectively sterilizing Legionella. Therefore, the bactericidal substance of the present invention can be used as a bactericidal composition containing the bactericidal substance as an active ingredient, and this killing effect is remarkable in that it is selective. In addition, the water treatment agent added to the water in the environment where Legionella is prone to proliferate by circulating or using the water in the tank of the cooling water system or humidifier, etc., is secondary to Legionella disease. Expected to be used as a disinfectant and disinfectant to prevent contamination

  Hereinafter, preferred embodiments of the present invention will be described. Throughout this specification, it should be understood that the singular forms also include the plural concept unless specifically stated otherwise. Thus, singular articles (eg, corresponding articles and adjectives in other languages, such as “a”, “an”, “the” in the case of English) are subject to the plural concept unless otherwise stated. Should also be understood to include. In addition, it is to be understood that the terms used in the present specification are used in the meaning normally used in the art unless otherwise specified. Thus, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control.

(Definition)
Listed below are definitions of terms particularly used in the present specification.

As used herein, “Legionella” includes bacteria belonging to the genus Legionella. If the species is not specified, Legionella sp. (In this specification, in the case of a specific species, it may be indicated as L. ***). This bacterium is a generic name for eubacteria belonging to the genus Legionella, is a Gram-negative gonococcus, includes species that cause many Legionella diseases, such as Legionella pneumonia (also called local military disease), and about 50 species, 70 serotypes are known facultative intracellular parasitic bacteria. Legionella is an aerobic gram-negative bacillus of about 2-5 μm and has one or more flagella. Legionella is facultative intracellular parasites, and in these places, it parasitizes the cells of other organisms such as protozoa such as amoeba, or coexists with algae, which allows it to grow in various environments. It is said that it is possible. When an aerosol containing Legionella is inhaled by humans, Legionella reaches the alveoli, where it is phagocytosed by alveolar macrophages. However, Legionella is facultative intracellular parasitic and can escape its bactericidal mechanism and grow in macrophages. Legionella are bacteria that are normally present in the environment and are usually less likely to cause infections. However, in the environment as shown in the environment prone to infection, especially for people with low resistance such as elderly people, a kind of Legionella genus, L. pneumophila causes human Legionella infections (Legionella pneumonia and Pontiac fever).

  The bacterium targeted by the present invention is not particularly limited as long as it is a Legionella bacterium, and it is understood that any bacterial species that requires treatment falls within the range. Specifically, Legionella pneumophila (eg, serum group 1, serum group 2, serum group 3, serum group 4, serum group 5, serum group 6, serum group 7, serum group 8), Legionella longumachee, Legionella michidea , Legionella oakridgensis, Legionella feelei, Legionella anisa, Legionella sainthelensi, Legionella bozemanii, Legionella gormanii, Legionellawell It can be directed to a la gormanii like.

  As used herein, “ribosyl derivative” such as “anthranilic acid” refers to anthranilic acid or a derivative thereof (eg, a fluoro derivative (ie, a derivative in which one or several hydrogen atoms are substituted with fluorine in anthranilic acid, for example, , 3-fluoroanthranilic acid, 4-fluoroanthranilic acid, 5-fluoroanthranilic acid, 6-fluoroanthranilic acid, etc.), and the amino group of anthranilic acid is substituted with ribose. Examples thereof include rate, ribosyl 3-fluoroanthranylate, ribosyl 4-fluoroanthranilate, ribosyl 5-fluoroanthranilate, and ribosyl 6-fluoroanthranilate.

As used herein, “deoxyribulose derivative” such as “anthranilic acid” refers to anthranilic acid or a derivative thereof (for example, 3-fluoroanthranilic acid, 4-fluoroanthranilic acid, 5-fluoroanthranilic acid, 6-fluoroanthranilic acid, etc. ) In which the amino group of anthranilic acid is substituted with deoxyribulose. For example, carboxyphenylaminodeoxyribulose (CPADR), 3-fluorocarboxyphenylaminodeoxyribulose, 4-fluorocarboxyphenylaminodeoxyribulose, 5-fluorocarboxyphenylaminodeoxyribulose, 6-fluorocarboxyphenylaminodeoxyribulose, etc. be able to.

  As used herein, “phosphor” refers to anthranilic acid and fluoro derivatives thereof, and those obtained by binding a phosphate group to ribose derivatives and deoxyribulose derivatives thereof. One phosphate group may be bonded, or a plurality of, for example, two, three, etc., may be bonded.

The compounds that are active ingredients herein include salts, preferably pharmaceutically acceptable salts. For example, alkali metals (such as lithium, sodium or potassium), alkaline earth metals (such as magnesium or calcium), ammonium, salts with organic bases and amino acids, or inorganic acids (hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, phosphorus Acid or hydroiodic acid)
And organic acids (acetic acid, trifluoroacetic acid, citric acid, lactic acid, tartaric acid, oxalic acid, maleic acid, fumaric acid, mandelic acid, glutaric acid, malic acid, benzoic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid , Methanesulfonic acid or ethanesulfonic acid, etc.). In particular, hydrochloric acid, phosphoric acid, tartaric acid, methanesulfonic acid and the like can be mentioned. These salts can be formed by a commonly performed method.

  As used herein, “solvate” refers to a complex of various stoichiometric amounts formed by a solute (eg, a compound used in the present invention) and a solvent, for example, a solvate with an organic solvent. Hydrates and the like. When forming a hydrate, it may be coordinated with any number of water molecules. Such solvents for the purposes of the present invention should not interfere with the biological activity of the solute. Examples of suitable solvents include but are not limited to water, methanol, ethanol and acetic acid.

  As used herein, “effective amount” means the amount of a drug or agent that elicits the biological or medical response of a tissue, system, animal or human of interest, for example, by those skilled in the art. . Furthermore, the term “therapeutically effective amount” refers to an improved treatment, cure, prevention or amelioration of a disease, disorder or side effect, or the rate of progression of a disease or disorder compared to a corresponding subject not receiving such an amount. Mean any amount that results in a decrease. The term also includes within its scope an amount effective to amplify normal physiological function.

  As used herein, “suppressing Legionella” means bacteriostatic of Legionella, that is, inhibition of bacterial growth.

  As used herein, “killing Legionella” means sterilization of Legionella, that is, killing Legionella.

  The term “antibacterial” as used herein refers to all negative effects on bacterial growth and growth, including bacteriostatic and bactericidal, when referring to Legionella. When described in detail, the “growth” of a bacterium refers to the increase of the bacterium, and the “growth” of the bacterium refers to continuing to live. The present invention may have an inhibitory action or an inhibitory action in any aspect, and is referred to as “growth inhibition” or simply “suppression” when inhibiting growth, and also referred to as “killing” when inhibiting “growth”.

  In the present specification, the term “antibacterial agent” has the broadest meaning used in the field, refers to a drug having an antibacterial action, a drug that causes destruction or growth inhibition of microorganisms and super microorganisms, and suppresses the growth of bacteria. In addition to drugs that have microbial growth, drugs that inhibit the growth of bacteria, bactericides in ground products, pharmaceuticals (for example, antibiotic preparations, etc.) are also included.

  As used herein, “a disease, disorder or condition associated with Legionella” refers to any disease, disorder or condition associated with Legionella. Legionella disease includes, for example, respiratory infection (Legionella pneumonia; veterans' disease) and Pontiac fever. Legionella-related disorders or conditions include those associated with Legionella pneumonia, starting with symptoms such as general malaise, headache, loss of appetite, muscle pain, and dry cough (after 2 to 3 days, purulent to reddish brown) ), High fever, chills, and chest pain. This disease is also characterized by the early appearance of central nervous system symptoms such as somnolence, coma, hallucinations, and tremor of the limbs. Related to Pontiac fever is its characteristic of starting with sudden fever, chills, and muscle pain, but healing temporarily. The present invention is not limited thereto, but can also treat or prevent diseases, disorders, symptoms and conditions related to Legionella.

  As used herein, “further antibacterial substance” means any antibacterial substance other than the antibacterial agent of the present invention. Therefore, such an antibacterial substance may be an antibacterial substance against Legionella bacteria or an antibacterial substance other than Legionella bacteria. Such antibacterial substances include β-lactams (eg, ampicillin, ceftazidime, meropenem), quinolones (eg, norfloxacin, ciprofloxacin), glycopeptides (eg, vancomycin), macrolides (eg, Erythromycin), oxazolidinones (eg, linezolid), peptide antibiotics (eg, magainin II), lipopeptides (eg, polymyxin, bacitracin), nitroimidazoles (eg, metronidazole), ansamycins (eg, rifampin) Azoles (eg, fluconazole), D-cycloserine, lincosamides (eg, clindamycin), mupirocin, streptogramins (eg, dalfopristin, quinupristin), fosfomycin, amino Synthesis of noglycosides (eg, gentamicin), sulfonamides (eg, sulfomethoxazole), trimethoprim, tetracyclines (eg, tigylcycline), novobiocin, chloramphenicol, monobactams, and antibiotics thereof Derivatives and the like can be mentioned. Further antibacterial substances include not only antibiotics but also general antibacterial alcohols such as ethanol and isopropanol, organic acid, hypochlorous acid, inorganic chlorine-based disinfectants represented by chlorine dioxide, It is understood that any antibacterial substance may be used as long as it has an antibacterial activity, including silver ion, photocatalytic antibacterial substance titanium oxide, zinc oxide, and the like. An antibacterial substance can be selected appropriately.

(Description of Preferred Embodiment)
The description of the preferred embodiment is described below, but it should be understood that this embodiment is an illustration of the present invention and that the scope of the present invention is not limited to such a preferred embodiment. It should be understood that those skilled in the art can easily make modifications, changes, and the like within the scope of the present invention with reference to the following preferred embodiments and demonstration examples in Examples.

(Legionella antibacterial agent)
In one aspect, the present invention provides the following formula:

and

Provided is an antibacterial agent against Legionella, comprising at least a compound selected from the group consisting of a compound having (fluoro derivative), a phospho form thereof, and a ribosyl derivative and a deoxyribulose derivative thereof.

  Here, the fluoro derivative is

Or a ribosyl derivative or a deoxyribosyl derivative thereof. These compounds were found to have antibacterial activity against Legionella when tested definitively. The following representative structural formulas are shown.

  The ribose derivative of the present invention can also be expressed as follows (from left: ribose derivative of anthranilic acid and its fluoro derivative).

  The deoxyribulose derivative of the present invention can also be expressed as follows (from the left, deoxyribulose derivative of anthranilic acid and its fluoro derivative).

  Since the compound of the present invention has the property of being easily phosphorylated in a living body such as a bacterium, it is presumed that the phosphorylated form of the compound also exhibits antibacterial activity. It is understood that the phospho form is also within the scope of the present invention.

  In the present invention, the compound that is the active ingredient of the present invention can be used at any concentration that inhibits the growth of Legionella, and can be used usually at a concentration that inhibits the growth. Including but not limited to a final concentration of 01 mM, such as at least 0.01 μM, at least 0.1 μM, at least 1 μM, or at least 0.02 mM, at least 0.03 mM, at least 0.04 mM, at least 0.05 mM, It can be at least 0.06 mM, at least 0.07 mM, at least 0.08 mM, at least 0.09 mM, at least 0.1 mM, or any value in between. On the other hand, the compound which is an active ingredient of the present invention is 10 mM or less, or 9 mM or less, 8 mM or less, 7 mM or less, 6 mM or less, 5 mM or less, 4 mM or less, 3 mM or less, 2 mM or less, 1 mM or less, or any between them Can be used. Legionella kills or inhibits growth, but can be used when it is desired to use it at a concentration that does not kill or inhibit growth of other representative bacteria.

In one embodiment, for example, 0.1 mM to 10 mM, 1.0 mM to 10 mM, 0.1
Although it can use in the range of mM-1 mM etc., it is not limited to these. This is because it was demonstrated in the Examples that killing or growth inhibition of Legionella was observed in this concentration range, and killing or growth inhibition of other representative bacteria was not observed. It is understood that the present invention is not limited to this because it may be used as a specific or selective antibacterial agent. In representative embodiments, it is understood that the lower limit may be 0.01 mM or more, and the upper limit may be 10 mM or less, or 10 mM or more.

  In one embodiment, the present invention can preferably use anthranilic acid or a fluoro derivative thereof as the active ingredient. Although not wishing to be bound by theory, anthranilic acid or its fluoro derivatives are more stable than their ribosyl derivatives.

In one embodiment, the antimicrobial agent of the present invention may contain additional antimicrobial substances. Since the antibacterial agent of the present invention has high specificity for Legionella bacteria and low killing ability or growth inhibition ability against other bacteria, selective killing or growth inhibition against Legionella bacteria can be performed. And if you want to kill other bacteria, mix antibacterial substances that have spectrum in the bacteria with the antibacterial agent of the present invention to produce dual specific antibacterial agents against Legionella and other bacteria. Can do. Such antibacterial agents are advantageous when there are bacteria that are desired to survive. For example, in the present invention, it has been demonstrated that 10 mM has no effect on Lactobacillus (Lactobacillus plantarum), which is a useful bacterium. Therefore, other lactic acid bacteria such as Lactobacillus acidophilus, Lactococcus (eg Lact
ococcus lactis etc.) can be mentioned as useful bacteria.

(Fungicide)
The present invention can be used as a disinfectant. Examples of the disinfectant include liquid products such as a disinfectant spray, powder, tablets (tablets), and the like.

  Such a bactericidal agent can be prepared by dissolving anthranilic acid, which is an active ingredient of the present invention, in an appropriate medium. Examples of media that can be used include water, organic solvents, alcohols, lipids, and fatty acids.

  This can be filled into an appropriate container. For such applications, any container used in the art can be used. If it is a spray, an aluminum can and a steel can can be used, and it can be filled with dimethyl ether, carbon dioxide, chlorofluorocarbon alternative, etc.

(Method of sterilization or bacteriostasis)
The present invention provides a method of inhibiting or killing Legionella. This method includes the step of contacting the antimicrobial agent of the present invention with Legionella. This contact can be achieved by, for example, spraying or applying the composition containing the antibacterial agent of the present invention and an appropriate medium as needed to a place where Legionella bacteria that have been suppressed or killed are present. In addition, the powder sterilizer can be used for sterilization of vomits and water tanks (tubs) with or without adding an appropriate excipient according to the purpose. A tablet-like disinfectant can be formed into a tablet shape by adding an appropriate excipient according to the purpose, and after controlling the sustained release, a continuous disinfecting effect can be expected by administering it to circulating water.

  Add antibacterial properties to coating agents (inorganic or organic), waxes, etc. by adding liquid disinfectants and powder disinfectants to paints, coating agents (inorganic or organic), waxes, etc. that match those characteristics. Can do. ). Also, in one embodiment, the contacting is performed in the presence of less than 0.7M, or less than 0.9M sodium chloride (NaCl). This is because at concentrations higher than these concentrations, bacteriostatic or bactericidal effects against other bacteria may be observed (Non-patent Document 1). If this NaCl concentration is a salt that affects other osmotic pressures, the same bacteriostatic or bactericidal effect may be seen. Therefore, when selectivity is considered important in the antibacterial agent of the present invention. It is advantageous to avoid such concentrations. On the other hand, when other bacteria need to be killed, a salt that affects the osmotic pressure, such as NaCl, can be added to make a fungicide. Examples of the salt having an influence on the osmotic pressure include potassium chloride (KCl) other than sodium chloride. In addition, examples of other bacteria include P.I. aeruginosa, B. et al. thailandensis, Y.M. pseudotuberculosis, S. p. aureus, B. et al. Examples include subtilis (Non-Patent Document 1).

(Articles coated with antibacterial agents)
The present invention provides an article coated or coated with the antibacterial agent of the present invention. Such an article can be produced by directly applying the antibacterial agent of the present invention to the article, or mixing it with an appropriate base and applying the medium to the intended article. As such a base, any widely used base such as paint, inorganic / organic coating agent, wax and the like can be used. For example, acrylic polymer, polyethylene glycol, silicon And a coating agent. As such articles, typically, articles in the field of hygiene products, such as containers, bath tubs, handrails, toothbrushes, air conditioner filters, masks, bathroom tiles, shower heads, bath tubs, deck brushes, hoses , A 24-hour bath circulator filter, a nebulizer, a humidifier tank, a curtain, a sheet (bedding), and the like. As for the combination of the base and the article, a person skilled in the art can select an appropriate combination in consideration of a known technique in consideration of wettability and the like. Examples of the resin coating agent or organic coating agent include urethane, fluororesin, polyethylene, nylon, and epoxy resin. Examples of the inorganic coating agent include a silicon coating agent, a titanium oxide coating agent, a silver coating agent, and a ceramic coating agent. Examples of the wax include paraffin wax (petroleum wax) and synthetic wax. Moreover, the coating agent can use the coating-type thing and the liquid thing used for a spray in this invention from the durability. Examples of the paint include water-based paint, oil-based paint, and powder paint. It is understood that the paint is a mixture and can include the substances listed as the coating agent.

(Treatment / Prevention Administration and Composition)
The present invention provides methods for the treatment, inhibition and prevention of Legionella related diseases, disorders or conditions by administration of an effective amount of a component or pharmaceutical composition comprising a compound of the present invention to a subject. In a preferred aspect, the component comprising the compound can be substantially purified (eg, substantially free of substances that limit its effect or cause undesirable side effects). The subject is preferably an animal, including but not limited to animals such as cows, pigs, horses, chickens, cats, dogs, and the like, and is preferably a mammal, and most preferably a human.

  Where the compounds of the invention are used as medicaments, such compositions may further comprise a pharmaceutically acceptable carrier and the like. Examples of the pharmaceutically acceptable carrier contained in the medicament of the present invention include any substance known in the art.

  Such suitable formulation materials or pharmaceutically acceptable carriers include antioxidants, preservatives, colorants, flavors, and diluents, emulsifiers, suspending agents, solvents, fillers, bulking agents, buffers. Include, but are not limited to, agents, delivery vehicles, diluents, excipients and / or pharmaceutical adjuvants. Typically, the medicament of the present invention is a composition comprising isolated pluripotent stem cells, or a variant or derivative thereof, together with one or more physiologically acceptable carriers, excipients or diluents. It is administered in the form of For example, a suitable vehicle can be water for injection, physiological solution, or artificial cerebrospinal fluid, which can be supplemented with other materials common to compositions for parenteral delivery. .

  Acceptable carriers, excipients or stabilizers used herein are non-toxic to the recipient and are preferably inert at the dosages and concentrations used, for example Phosphate, citrate, or other organic acids; ascorbic acid, α-tocopherol; low molecular weight polypeptide; protein (eg, serum albumin, gelatin or immunoglobulin); hydrophilic polymer (eg, polyvinylpyrrolidone); amino acid (Eg, glycine, glutamine, asparagine, arginine or lysine); monosaccharides, disaccharides and other carbohydrates (including glucose, mannose, or dextrin); chelating agents (eg, EDTA); sugar alcohols (eg, mannitol or sorbitol) Salt formation vs. Io (E.g., sodium); and / or non-ionic surface-active agents (e.g., Tween, Pluronic (pluronic) or polyethylene glycols (PEG)) but the like are not limited thereto.

Exemplary suitable carriers include neutral buffered saline or saline mixed with serum albumin. Preferably, the product is formulated as a lyophilizer using a suitable excipient (eg, sucrose). Other standard carriers, diluents and excipients may be included as desired. Other exemplary compositions are Tri-pH 7.0-8.5.
s buffer or acetate buffer at pH 4.0-5.5, which may further include sorbitol or a suitable substitute thereof.

  The medicament of the present invention can be administered orally or parenterally. Alternatively, the medicament of the present invention can be administered intravenously or subcutaneously. When administered systemically, the medicament used in the present invention may be in the form of a pharmaceutically acceptable aqueous solution free of pyrogens. Such a pharmaceutically acceptable composition can be easily prepared by those skilled in the art by considering pH, isotonicity, stability and the like. In this specification, the administration method includes oral administration, parenteral administration (for example, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, mucosal administration, intrarectal administration, intravaginal administration, local administration to the affected area, Skin administration, etc.). Formulations for such administration can be provided in any dosage form. Examples of such a pharmaceutical form include a liquid, an injection, and a sustained release agent.

  Other dosage forms may be solid, semi-solid, liquid, etc., for internal or external use, such as tablets, capsules, solutions, vapors, ointments, pastes, sprays and the like.

  The medicament of the present invention may be a physiologically acceptable carrier, excipient or stabilizer (Japanese Pharmacopoeia 15th edition, its supplement or its latest edition, Remington's Pharmaceutical Sciences, 18th Edition, A R. Gennaro, ed., Mack Publishing Company, 1990, etc.) and a compound having the desired degree of purity can be prepared and stored in the form of a lyophilized cake or aqueous solution.

  The amount of the composition used in the treatment method of the present invention takes into consideration the purpose of use, the target disease (type, severity, etc.), the patient's age, weight, sex, medical history, cell morphology or type, etc. Can be easily determined by those skilled in the art. The frequency with which the treatment method of the present invention is applied to a subject (or patient) also depends on the purpose of use, target disease (type, severity, etc.), patient age, weight, sex, medical history, treatment course, etc. In view of this, it can be easily determined by those skilled in the art. Examples of the frequency include administration every day to once every several months (for example, once a week to once a month). It is preferable to administer once a week to once a month while observing the course.

  The present invention can also provide the active ingredient of the present invention as a physiologically functional derivative. As used herein, “physiologically functional derivative” refers to a pharmaceutically acceptable derivative of any of the compounds of the present invention, such as an ester or amide, which when administered to a mammal A derivative that can provide (directly or indirectly) a compound of the invention or an active metabolite thereof. Such derivatives can be produced without undue experimentation by Burger's Medicinal Chemistry And Drug Discovery, 5th edition, Vol 1: Principles and Practice (the content of this document is the scope of teaching physiologically functional derivatives). And will be apparent to those skilled in the art with reference to the teachings of

  References such as scientific literature, patents and patent applications cited herein are hereby incorporated by reference in their entirety to the same extent as if each was specifically described.

  The present invention has been described with reference to the preferred embodiments for easy understanding. In the following, the present invention will be described based on examples, but the above description and the following examples are provided only for the purpose of illustration, not for the purpose of limiting the present invention. Accordingly, the scope of the present invention is not limited to the embodiments or examples specifically described in the present specification, but is limited only by the scope of the claims.

  Reagents used in the following examples were obtained from Wako Pure Chemical Industries, Ltd., Sigma Aldrich Japan Co., Ltd., Tokyo Kasei Kogyo Co., Ltd., Kanto Chemical Co., Ltd., etc. Obtained or available from RIKEN BioResource Center, American Type Culture Collection, etc. Alternatively, it was synthesized by commissioning to Nard Laboratories.

(Example 1: Antibacterial activity of anthranilic acids against Legionella bacteria)
In this example, the antibacterial activity of anthranilic acids against Legionella was examined.

(Method and material)
(Substances used; abbreviations in parentheses are used. Also, the following parentheses are the manufacturing company and its catalog number.)
Anthranilic acid (AA) (obtained from Kanto Chemical Co., Inc., 0141-30)
4-fluoroanthranilic acid (4-FAA) (obtained from Tokyo Chemical Industry Co., Ltd., F0405)
5-fluoroanthranilic acid (5-FAA) (obtained from Wako Pure Chemical Industries, Ltd., 322-82431)
6-fluoroanthranilic acid (6-FAA) (obtained from Wako Pure Chemical Industries, Ltd., 328-66651)
Methyl anthranilate (AM) (obtained from Wako Pure Chemical Industries, Ltd., 138-01973) Methyl N-methyl anthranilate (MAM) (obtained from Wako Pure Chemical Industries, Ltd., 134-04253)
3-Chloroanthranilic acid (4-ClAA) (obtained from Wako Pure Chemical Industries, Ltd., 328-55421) 1.0 mM
4-Chloroanthranilic acid (4-ClAA) (obtained from Tokyo Chemical Industry Co., Ltd., A0661) 1.0 mM
2-amino-5-chlorobenzoic acid (5-ClAA) (5-chloroanthranilic acid) (obtained from Wako Pure Chemical Industries, 321-50391) 1.0 mM
2-Amino-6-chlorobenzoic acid (6-ClAA) (6-chloroanthranilic acid) (obtained from Wako Chemical, 321-224761) 1.0 mM
Carbonylphenylaminodeoxyribosylate (CPADR) (manufactured by Nard Laboratories)
Chorismic acid (CA) (obtained from Sigma-Aldrich Japan, C1761)
Indole (obtained from Wako Pure Chemical Industries, 093-00162)
L-tryptophan (L-Trp) (obtained from Peptide Institute, 2721)
Benzoic acid (BA) (obtained from Kanto Chemical Co., Ltd., 04115-30)
Ribosyl anthranilic acid (RAA) (obtained by combining AA and ribose (Nard Laboratories))
Ribosyl 4-fluoroanthranilic acid (RFAA) (obtained by combining FAA and ribose (Nard Laboratories))
Ribosylanthranilic acid benzyl ester (RAABn) (Nard Research Laboratories)
Ribosyl 4-chloroanthranilic acid (RClAA) (Nard Research Laboratories)
Ribosyl-dimethylanthranilic acid (RdMAA) (Nard Research Laboratories)
p-aminobenzoic acid (p-ABA) (obtained from Wako Pure Chemical Industries, Ltd., 015-02332)
L-kynurenine (LK) (obtained from Sigma-Aldrich Japan, K8625)
Picolinic acid (P) (obtained from Sigma-Aldrich Japan, P42800).

  (Tested strain)

  These bacterial strains were inoculated with 1 platinum loop of microbial cells in a fresh LS medium and cultured at 35 ° C. and 150 rpm.

(Plate assay medium preparation)
(Basic medium)
Yeast extract (Merck 1.03753) 10.0 g
Gellan gum (Wako Pure Chemicals 073-03071) 10.0g
Magnesium chloride hexahydrate (Wako Pure Chemicals 135-09205) 0.1g / 900mL
These were mixed and autoclaved at 121 ° C. for 15 minutes.

(Growth supplements)
ACES (N- (2-acetamido) -2-amino-ethanesulfonic acid) (Dojindo 349-04881) 10.0 g
Iron (III) diphosphate, soluble (iron (III) pyrophosphate, soluble) (Wako Pure Chemical Industries, Ltd. 090-02405) 0.250 g
L-cysteine (Wako Chemicals 322-20612) 0.40 g
2-Oxoglutaric acid (α-ketoglutaric acid) (Wako Pure Chemicals 119-00085) 1.0 g
The pH was 6.90 / 100 mL.

The growth supplement was sterilized with a filter and then added to the medium and mixed. After 21 hours, the pouch strain culture was terminated. For Legionella, L prepared a pour medium using the stock solution.
Since other strains grew faster than Legionella, a pour medium was prepared using a 10-fold diluted solution. 0.2 mL of the culture solution or diluted solution was added to 10 mL of the agar medium cooled to 55 ° C., and the above was mixed in a sterile petri dish to prepare a pour plate.

(Plate assay)
The plate assay was performed as follows.

  Each assay sample was dissolved in 100% or 50% acetonitrile to prepare an appropriate concentration solution, and 2.0 μL of the assay sample was spotted on each pour plate. The numbers in the table indicate the diameter (mm) of the growth inhibition circle. -Indicates not implemented.

(result)
The results of this example are summarized in the following table.

  (Anthranilic acid and its derivatives)

  (Anthranilic acid analog)

  (Ribosylanthranilic acid derivative and CPADR)

In three types of AA and AA fluoro derivatives, growth inhibition circles were observed around the sample spot in Legionella bacteria. For fluoro derivatives, inhibitory activity is observed at the 4th, 5th and 6th positions tested. The chloro derivative has no inhibitory activity regardless of the 3-position, 4-position, 5-position and 6-position. In other strains, it was found that no growth inhibitory activity was observed under this assay condition.

  From the above, the growth inhibitory effect on Legionella at the present time was confirmed to be anthranilic acid (AA), 4-fluoroanthranilic acid, 5-fluoroanthranilic acid, 6-fluoroanthranilic acid, ribosyl 3-fluoroanthranilic acid, It can be said that ribosyl 4-fluoroanthranilic acid, ribosyl 5-fluoroanthranilic acid, ribosyl 6-fluoroanthranilic acid, and carbonylphenylaminodeoxyribosylate have Legionella killing effect. A similar effect is expected for 3-fluoroanthranilic acid.

  On the contrary, no activity is observed when chorismate (CA), indole (Ind), tryptophan (Trp), benzoic acid (BA), AA derivatives (methyl anthranilate (AM), methyl N-methylanthranilate (MAM) ), 4-chloroanthranilic acid (ClAA)), ribosyl anthranilic acid benzyl ester (RAABn), ribosyl 4-chloroanthranilic acid (RClAA), ribosyl-dimethylanthranilic acid (RdMAA)).

  In lactic acid bacteria (Lp), neither anthranilic acid nor CPADR showed growth inhibitory effects. Therefore, it became clear that there was specificity for Legionella and that only Legionella can be killed without killing lactic acid bacteria, which are useful bacteria.

  (Other related chemical substances)

  K and P are substances (Koji N et al Bio. Pharm. Bull 2009 32; 41-44) that have been investigated to have antibacterial activity against multi-drug resistant staphylococci and the like. Expected to be active. However, no activity against Legionella was found. Moreover, although p-ABA is an isomer of anthranilic acid (o-ABA), antibacterial activity was not recognized.

(Example 2: Strength of growth inhibitory activity of each substance)
About AA, FAA, CPADR in which the growth inhibitory activity was confirmed as described above, the strength of the growth inhibitory activity was verified by changing the concentration of the spotted sample. Each sample was serially diluted with 50% acetonitrile, spotted on an assay plate mixed with Legionella bacteria, 2 μL each, and cultured overnight, and a growth inhibition circle appearing around the spot was observed.

  As shown in the above table, there was no significant difference in the strength of the growth inhibitory activity among the three substances AA, FAA, and CPADR. Also, the minimum inhibitory concentration (MIC) appears to be between 0.1 mM and 0.01 mM. Here, it is recognized that Δ and ○ have the antibacterial activity referred to in the present invention.

(Example 3: Verification of growth inhibitory activity in Legionella pour plate <Verification of growth inhibitory effect in other bacterial species>)
In this example, the growth inhibitory effect of other strains was verified. That is, similar to the method described in Example 1, the growth inhibitory effect of Legionella and other bacterial species on the AA and CPADR-added plates was verified. A plate to which AA and CPADR were added was prepared, Legionella and other bacterial species were applied to the plate, and the growth inhibitory effect of each bacterial species on AA was verified.

(Strain used; abbreviations in parentheses)
Legionella: Legionella pneumophila subsp. pneumophila JCM7571 (L)
Other species:
Bacillus subtilis ATCC6633 (Bs)
Escherichia coli NBRC 3972 (Ec)
Pseudomonas aeruginosa NBRC13275 (Pa)
Staphylococcus aureus NBRC13276 (Sa)
In these, cells of one platinum loop were inoculated into a fresh LS (Linsmeier and Skog) medium, and culture was started at 35 ° C. and 150 rpm.

(Medium preparation)
(Basic medium)
Yeast extract (Merck 1.03753) 6.0g
Gellan gum (Wako Pure Chemicals 073-03071) 6.0g
Magnesium chloride hexahydrate (Wako Pure Chemicals 135-09205) 0.06g / 540mL
These were prepared by autoclaving at 121 ° C. for 15 minutes.

(Growth supplements)
ACES (N- (2-acetamido) -2-amino-ethanesulfonic acid) (Dojindo 349-04881) 6.0 g
Iron (III) diphosphate, soluble (iron (III) pyrophosphate, soluble) (Wako Pure Chemical Industries, Ltd. 090-02405) 0.150 g
L-cysteine (Wako Chemicals 322-20612) 0.24 g
2-Oxoglutaric acid (α-ketoglutaric acid) (Wako Pure Chemicals 119-00085) 0.6 g
The pH was 6.90 / 60 mL.

  These were added to and mixed with the medium after filter sterilization.

(Assay)
AA and CPADR were added to a sterile petri dish as follows.

10 mL of the above medium was dispensed into each dish and mixed with AA and CPADR to solidify.

(Plate assay)
After 17 hours, the culture was terminated. Each culture solution was diluted 10 ³ to 10 ⁶ times with a phosphate buffer. Next, 50 μL of each of the above dilutions was applied to the AA and CPADR-added plates, and 50 μL of each of the same dilutions were applied to the NC plate. The culture was started at 35 ° C. The result was evaluated by counting the number of bacteria on the plate.

(Plate check result)
The results are shown below.

The indication (small) in the table indicates that colonies are formed, but the colony diameter is very small compared to the others.

L was found to be about 10 times more sensitive than AA and CPADR. As for other bacteria, it was found that when 10 mM AA was added to Bs, colonies were not formed at all, and when 1 mM CPADR was added, only a recovery rate of about 50% without addition was obtained. It was also found that Pa was not inhibited by 10 mM AA or 1 mM CPADR, and Ec was colony grown even when 10 mM AA was added, but the colony was considerably small and was inhibited to some extent. .
It was also found that 1 mM CPADR addition did not inhibit growth at all. It was found that Sa showed almost the same result as Ec.

  From the above, it was found that the antibacterial agent of the present invention is specific or selective to Legionella bacteria compared to these bacteria.

(Example 4: Implementation with spray product)
In this embodiment, an example is shown in which the spray product is used.

  The antibacterial agent of the present invention is dissolved in a normal sterilizing spray containing 70% ethanol as a main component, and the antibacterial action is further enhanced against Legionella bacteria.

(Example 5: Implementation in a bath)
In the present embodiment, an example when the bath pot product is used is shown. When molding the bath, the antibacterial agent of the present invention is kneaded so that Legionella bacteria hardly grow on the surface.

(Example 6: Implementation with a circulating bath filter)
In this embodiment, an example is shown in which the filter is used in a circulating bath filter.

  The antibacterial agent of the present invention is kneaded into a circulating bath kettle filter to make it difficult for Legionella to grow on the surface.

  As mentioned above, although this invention has been illustrated using preferable embodiment of this invention, it is understood that the scope of this invention should be construed only by the claims. Patents, patent applications, and documents cited herein should be incorporated by reference in their entirety, as if the contents themselves were specifically described herein. Understood.

  The present invention provides a novel antibacterial agent against Legionella. It has been found that the present invention can provide an antibacterial agent with a compound having a relatively simple structure and also exhibits selectivity. Because of such effects, the present invention is useful in industries where antibacterial agents are used, and finds use in the pharmaceutical industry.

Claims (14)

The following formula:
and
And an antibacterial agent against Legionella, comprising at least a compound selected from the group consisting of a ribosyl derivative and a deoxyribulose derivative, or a phospho form, a salt or a solvate thereof. The antibacterial agent according to claim 1, wherein the compound is contained at a concentration of at least 0.01 mM. The antibacterial agent according to claim 1, wherein the compound is contained at a concentration of 0.1 mM to 10 mM. The antibacterial agent according to claim 1, wherein the compound is contained at a concentration of 1.0 mM to 10 mM. The compound is
Or
The antibacterial agent according to claim 1, wherein The antibacterial agent according to any one of claims 1 to 6, further comprising an antibacterial substance. The antibacterial agent according to claim 1, which is liquid. A pharmaceutical composition comprising the antibacterial agent according to claim 1. An article comprising the antibacterial agent according to claim 1. The article according to claim 9, wherein the antibacterial agent is coated on the article. The article according to claim 9, wherein the article is selected from a container, a bath tub, a handrail, a toothbrush, a filter, a mask, a tile, a shower head, a bath tub, a deck brush, a hose and a nebulizer. The article according to claim 9, wherein the antibacterial agent includes at least one selected from the group consisting of a paint, an inorganic coating agent, an organic coating agent, and a wax as a base. A method for inhibiting or killing Legionella, comprising the step of bringing the antibacterial agent according to any one of claims 1 to 7 into contact with the Legionella in an effective amount for inhibiting or killing. 14. The method of claim 13, wherein the contacting is performed in the presence of less than 0.7M sodium chloride (NaCl).