JP2017105714A - Multiple drug discharge pump inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a compound having the activity to inhibit the discharge ability of a multiple drug discharge pump, and a pharmaceutical composition comprising the compound as an active ingredient.SOLUTION: The present invention provides a compound represented by general formula (I) [where Ris a thiazole group or an adamantyl group in which one hydrogen atom is optionally substituted with an alkyl group having 1 to 4 carbon atoms, or an alkyl group having 3 to 6 carbon atoms, X is a hydrogen atom or a halogen atom, Ris an amino group or a hydrogen atom, n is an integer of 1-4, Ris an amino group or an imidazole group in which one hydrogen atom is optionally substituted with an amidino group] or its physiologically acceptable salt, or their solvates.SELECTED DRAWING: None

Description

  The present invention relates to compounds useful for the prevention or treatment of bacterial infections, particularly multidrug resistant bacterial infections, and more specifically, the ability to inhibit the drainage ability of multidrug efflux pumps that are highly expressed in multidrug resistant bacteria. And a pharmaceutical composition containing the compound as an active ingredient.

  Many antibacterial agents have been developed to date for the prevention and treatment of infections caused by bacteria, and β-lactams, aminoglycosides, quinolones, macrolides, tetracyclines, and other compounds have been put into practical use as pharmaceuticals. I came. However, in recent years, the emergence of resistant bacteria to these antibacterial agents has become a serious problem in clinical practice. Among infections caused by drug-resistant bacteria, gram-negative multidrug-resistant bacteria such as multidrug-resistant Pseudomonas aeruginosa (MDRP), multidrug-resistant Acinetobacter (MDRA), and carbapenem-resistant enterococci (CRE) are methicillin-resistant Staphylococcus aureus ( Unlike gram-positive multidrug-resistant bacteria such as MRSA), there are few therapeutic agents that can be addressed. For this reason, nosocomial infections caused by Gram-negative multidrug-resistant bacteria are a major problem. A method of dealing with an infection caused by a gram-negative multidrug-resistant bacterium with a new antibacterial agent may lead to the emergence of a new resistant bacterium, and is unlikely to be a fundamental solution. In order to overcome multidrug-resistant gram-negative bacterial infections, the creation of prevention / treatment strategies based on new concepts is an urgent issue that is socially demanded.

  Bacterial multidrug resistance is caused by the acquisition of multiple resistance factors. Examples of the acquisition of the resistance factor include production of carbapenemase and accumulation of DNA gyrase mutation. While the individual resistance spectrum of these resistance factors is clear, recent multi-drug resistant bacteria are characterized by a very broad resistance to almost all antibiotics. Underlying such widespread multidrug resistance in gram-negative bacteria is the high expression of the multidrug efflux system. The multidrug efflux system of Gram-negative bacteria is a multicomponent efflux protein complex, and AcrAB-TolC in Escherichia coli, MexAB-OprM and MexXY-OprM in Pseudomonas aeruginosa are known. Among these, AcrB, MexB, and MexY are RND (resistance / nodulation / division) type multidrug efflux pumps that exist on the inner membrane of Gram-negative bacteria and excrete various drugs using proton driving force. (Non-Patent Document 1). Drug drainage by these multidrug pumps gains other resistance factors in low drug environments in addition to the direct effect of lowering drug concentration in the bacterial body and significantly reducing drug sensitivity It has the indirect effect of giving the opportunity to do so, and has become a factor for the multidrug resistance of bacteria.

  Based on the above background, the concept of a concomitant drug that makes existing antibacterial agents effective by inhibiting the discharge ability of the multidrug discharge pump, which is the cause of multidrug resistance, has been submitted and developed. It was. For example, it has been reported that ABI-PP (D13-9001) having pyridopyrimidine as a nucleus inhibits the excretion ability of AcrB of E. coli and MexB of Pseudomonas aeruginosa and increases the susceptibility of bacteria to known antibacterial agents. (Patent Document 1). In addition, an oligopeptide compound (PAβN) in which phenylalanine, arginine, and 3-aminoquinoline are linked by an amide bond and related compounds (Patent Documents 2 to 5), and a group of compounds having an amidine group at both ends of the molecule (pentamidines) (Patent Documents 6 and 7) are reported to reduce the minimum inhibitory concentration (MIC) of known antibacterial agents against Pseudomonas aeruginosa strains in which the multidrug efflux pump is highly expressed. However, none of these compounds has been applied to medicine, and to date, there has been no application of a multidrug excretion inhibitor as a medicine.

  In recent years, rational drug molecular design (rational drug design) has been recognized as an important approach in drug discovery. In particular, three-dimensional structure information of a protein serving as a drug target is clarified by crystal structure analysis, and pharmaceutical design (SBDD) based on the three-dimensional structure information is actively attempted as a highly accurate method. For SBDD, it is necessary to obtain a co-crystal structure of a drug and a target protein, but it is difficult to obtain a crystal structure of a multidrug efflux pump that is a membrane protein. The inventors of the present invention succeeded in crystal structure analysis of AcrB for the first time in the world (Non-patent Document 2), and also achieved co-crystal structure analysis of AcrB with several known antibacterial agents discharged as AcrB substrates. For the first time in the world, we have succeeded in elucidating the drug discharge mechanism of a multidrug discharge pump at the molecular level (Non-patent Documents 3 and 4). Furthermore, the co-crystal structure analysis of the excretion inhibitor ABI-PP (D13-9001) and MexB was conducted to clarify the existence of the inhibitor binding pocket, and ABI-PP is another multidrug excretion pump of Pseudomonas aeruginosa. The structural factor that cannot inhibit the excretion ability of MexY is also clarified (Non-patent Document 5).

JP 2002-322004 A International Publication No. 2005/113579 US Patent Application Publication No. 2008/0076741 International Publication No. 2008/141010 International Publication No. 2008/141012 International Publication No. 2005/089738 US Patent Application Publication No. 2008/0132457

Yamaguchi et al., Frontiers in Microbiology, 2015, vol.6, Article 327. Murakami et al., Nature, 2002, vol.419, p.587-593. Murakami et al., Nature, 2006, vol.443, p.173-179. Nakashima et al., Nature, 2011, vol.480, p.565-569. Nakashima et al., Nature, 2013, vol.500, p.102-106. Nishino et al., Journal of Infection and Chemotherapy, 2008, vol.14, p.23-29.

  The present invention relates to a compound having an activity of inhibiting the discharge ability of a multidrug efflux pump, in particular, a compound having an efflux inhibitory activity against both of the two multidrug efflux pumps MexB and MexY possessed by Pseudomonas aeruginosa An object of the present invention is to provide a medicinal composition containing as an active ingredient.

The compounds, multidrug efflux pump inhibitors, and pharmaceutical compositions according to the present invention are the following [1] to [12].
[1] The following general formula (I)

[In Formula (I), R ¹ represents a thiazole group, an adamantyl group, or an alkyl group having 3 to 6 carbon atoms in which one hydrogen atom may be substituted with an alkyl group having 1 to 4 carbon atoms. X represents a hydrogen atom or a halogen atom, R ² represents an amino group or a hydrogen atom, n represents an integer of 1 to 4, and R ³ represents one hydrogen atom substituted with an amidino group. An amino group or an imidazole group which may be present. ]
Or a physiologically acceptable salt thereof, or a solvate thereof.
[2] R ¹ is a thiazole group in which one hydrogen atom may be substituted with an alkyl group having 1 to 4 carbon atoms, X is a hydrogen atom, a chlorine atom, or a bromine atom, and R ² Or a physiologically acceptable salt thereof, or a solvate thereof, wherein is an amino group.
[3] The compound of [1] or [2] above, wherein R ³ is an unsubstituted amino group or an amino group in which one hydrogen atom is substituted with an amidino group, and n is 3 or 4 or a compound thereof Physiologically acceptable salts or solvates thereof.
[4] The compound represented by the general formula (I) is represented by the following formulas (H-31), (H-32), (H-34) to (H-37), (H-39), (H -46), (H-47), (H-50), or (H-51)

The compound of any one of the above-mentioned [1] to [3], or a physiologically acceptable salt thereof, or a solvate thereof.
[5] The compound according to any one of [1] to [4] or a physiologically acceptable salt thereof, or one or more of these solvates as an active ingredient, A multidrug efflux pump inhibitor having an activity of inhibiting drug excretion ability.
[6] The multidrug efflux pump inhibitor according to the above [5], which has an activity of inhibiting both the drug excretion ability of MexB and the drug excretion ability of MexY.
[7] A pharmaceutical composition comprising one or more of the compounds according to any one of [1] to [4] or a physiologically acceptable salt thereof, or a solvate thereof as an active ingredient. .
[8] The pharmaceutical composition according to the above [7], which is used for prevention and / or treatment of bacterial infection.
[9] The pharmaceutical composition according to [8], further comprising one or more antibacterial agents.
[10] The pharmaceutical composition according to [8] or [9], wherein the pathogenic bacterium of the bacterial infection is a gram-negative bacterium.
[11] The pharmaceutical composition according to [8] or [9], wherein the pathogenic bacterium of the bacterial infection is Pseudomonas aeruginosa.
[12] The pharmaceutical composition according to any of [8] to [11], wherein the pathogenic bacterium of the bacterial infection is a bacterium that has acquired resistance to an antibacterial agent.

  The compound according to the present invention has an inhibitory ability against the RND type multidrug efflux pump possessed by gram-negative bacteria represented by MexB and MexY. Therefore, a pharmaceutical composition comprising the compound or a physiologically acceptable salt thereof or a solvate thereof as an active ingredient is a multidrug resistant gram-negative bacterial infection represented by multidrug resistant Pseudomonas aeruginosa. It is very useful for prevention or treatment.

FIG. 1 shows the results of measuring over time the OD ₆₀₀ of a culture medium in which each Escherichia coli strain was cultured in a culture medium in which H-31 compounds were added at various concentrations in Test Example 1. FIG. 2 shows the results of measurement over time of OD ₆₀₀ of a culture medium in which each Escherichia coli strain was cultured in a culture medium in which H-32 compounds were added at various concentrations in Test Example 1. FIG. 3 shows the results of measurement over time of OD ₆₀₀ of a medium in which each Escherichia coli strain was cultured in a medium to which H-34 compound was added at each concentration in Test Example 1. FIG. 4 shows the results of measuring over time the OD ₆₀₀ of a culture medium in which each Escherichia coli strain was cultured in a culture medium in which H-36 compound was added at various concentrations in Test Example 1. FIG. 5 shows the results of measurement over time of OD ₆₀₀ of a medium in which each Escherichia coli strain was cultured in a medium to which H-37 compound was added at each concentration in Test Example 1. FIG. 6 shows the results of measuring over time the OD ₆₀₀ of a culture medium in which each Escherichia coli strain was cultured in a culture medium in which H-31 compounds were added at various concentrations in Test Example 1. FIG. 7 is a graph showing the minimum growth inhibitory concentration (MIC) cumulative distribution when ciprofloxacin (CPFX) and H-31 compound are used in combination with clinically isolated MDRP55 strain in Test Example 2. FIG. 8 is a graph showing the cumulative distribution of MIC at the time of combined use of imipenem (IPM) and H-31 compound for clinically isolated MDRP55 strain in Test Example 2. FIG. 9 is a graph showing the cumulative distribution of MIC during combined use of amikacin (AMK) and H-31 compound for clinically isolated MDRP55 strain in Test Example 2. FIG. 10 is a graph showing the cumulative distribution of MIC during the combined use of Aztreonam (AZT) and H-31 compound for clinically isolated MDRP55 strain in Test Example 2.

  The known elimination inhibitor ABI-PP (D13-9001) has the ability to inhibit MexB among the two major multidrug efflux pumps of Pseudomonas aeruginosa, but cannot inhibit the ability to excrete MexY. (Non-patent document 5). The reason why ABI-PP cannot inhibit the excretion ability of MexY is that the pyridopyrimidine skeleton at the center of the molecule of ABI-PP is based on the result of co-crystal structure analysis of ABI-PP and MexB. It was presumed that it had to sterically interfere with the tryptophan residue present in the inhibitor binding pocket. Therefore, the present inventors must replace the bicyclic pyridopyrimidine skeleton at the center of the molecule of ABI-PP with a monocyclic skeleton in order to simultaneously inhibit the excretion ability of MexB and MexY. Based on the inference, a compound having an inhibitory ability against both MexB and MexY excretory ability was developed.

  The compound according to the present invention is a compound represented by the following general formula (I). The compound has a dual inhibitory ability to simultaneously inhibit the drug efflux ability of both MexB and MexY RND type multidrug efflux pumps. For this reason, the compound represented by general formula (I) can be used as an active ingredient of a multidrug efflux pump inhibitor.

In general formula (I), R ¹ represents a thiazole group, an adamantyl group, or an alkyl group having 3 to 6 carbon atoms. When R ¹ is an alkyl group having 3 to 6 carbon atoms, the alkyl group includes a propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a t-butyl group (tert-butyl group), and an n-pentyl group. , Isoamyl group, neopentyl group, n-hexyl group, isohexyl group, texyl group and the like.

When R ¹ is a thiazole group, it may be an unsubstituted thiazole group or a group in which one hydrogen atom in the thiazole group is substituted with an alkyl group having 1 to 4 carbon atoms. When one hydrogen atom in the thiazole group is substituted with an alkyl group, the alkyl group includes a methyl group, an ethyl group, a propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a t-butyl group ( tert-butyl group).

As the compound represented by the general formula (I), R ¹ is preferably a thiazole group because of its higher discharge capacity inhibitory activity against a multidrug discharge pump, and one hydrogen atom has 1 to 1 carbon atoms. More preferably, it is a thiazole group substituted with 4 alkyl groups, more preferably a thiazole group in which one hydrogen atom is substituted with an alkyl group having 3 to 4 carbon atoms, one hydrogen atom Is more preferably a thiazole group substituted with a t-butyl group.

  In general formula (I), X represents a hydrogen atom or a halogen atom. Examples of the halogen atom include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom. As the compound represented by the general formula (I), those in which X is a hydrogen atom, a chlorine atom or a bromine atom are preferable, and since X has a higher discharge capacity inhibiting activity for a multidrug discharge pump, X is a chlorine atom or bromine. What is an atom is more preferable, and what is a chlorine atom is still more preferable.

In general formula (I), R ² represents an amino group or a hydrogen atom. When R ² is an amino group, as the compound represented by the general formula (I), the configuration of the amino group of R ² may be (R) isomer or (S) isomer. The racemate may be used. As the compound represented by the general formula (I), a compound in which R ² is an amino group is preferable because it has a higher discharge capacity inhibitory activity for a multidrug discharge pump.

  In general formula (I), n represents an integer of 1 to 4.

In general formula (I), R ³ represents an amino group or an imidazole group in which one hydrogen atom may be substituted with an amidino group. When R ³ is an unsubstituted amino group, n is preferably 2, 3, or 4, more preferably 3 or 4. When R ³ is an amino group in which one hydrogen atom is substituted with an amidino group, n is preferably 2, 3, or 4, and more preferably 3 or 4. When R ³ is an imidazole group, n is preferably 1 or 2, and more preferably 1.

As the compound represented by the general formula (I), R ³ is an unsubstituted amino group or an amino group in which one hydrogen atom is substituted with an amidino group because of its higher excretion ability inhibiting activity against a multidrug efflux pump. And a compound in which n is 3 or 4 is preferred, a compound in which R ³ is an unsubstituted amino group and n is 3 or 4, or a hydrogen atom in R ³ is an amidino group A compound that is a substituted amino group and n is 3 is more preferable, and a compound that R ³ is an unsubstituted amino group and n is 4 is more preferable.

As the compound represented by the general formula (I), R ¹ is an unsubstituted thiazole group or a thiazole group in which one hydrogen atom is substituted with an alkyl group having 1 to 4 carbon atoms, and X is a hydrogen atom. , A chlorine atom, or a bromine atom, R ² is an amino group, n is an integer of 1 to 4, R ³ is an unsubstituted amino group, and one hydrogen atom is substituted with an amidino group A compound that is a group or an imidazole group, R ¹ is a thiazole group in which one hydrogen atom is substituted with an alkyl group having 1 to 4 carbon atoms, and X is a hydrogen atom, a chlorine atom, or a bromine atom R ² is an amino group, n is an integer of 2, 3, or 4, and R ³ is an unsubstituted amino group or an amino group in which one hydrogen atom is substituted with an amidino group. Preferably, R ¹ is one hydrogen atom being a t-butyl group A substituted thiazole group, X is a hydrogen atom, a chlorine atom, or a bromine atom, R ² is an amino group, n is 3 or 4, and R ³ is an unsubstituted amino group or one More preferred are compounds in which the hydrogen atom is an amino group substituted with an amidino group.

  More specifically, the compound represented by the general formula (I) is represented by the following formulas (H-31), (H-32), (H-34) to (H-37), (H-39). , (H-46), (H-47), (H-50), or (H-51).

The compound represented by the general formula (I) can be obtained, for example, by condensing an amino group of a benzamide derivative and a carboxylic acid in the presence of a condensing agent by a reaction shown in the following reaction formula (A). In the reaction formula (A), R ¹ , R ² , R ³ , X and n are the same as those in the general formula (I). In this reaction, the reactive group in the carboxylic acid is preferably protected in advance with a protecting group and deprotected after the condensation reaction.

For example, among the compounds represented by the general formula (I), R ¹ is a thiazole group in which one hydrogen atom is substituted with a t-butyl group, X is a hydrogen atom, a chlorine atom or a bromine atom, A compound in which R ² is an amino group is obtained by reacting 3-amino-N- (4-t-butyl-1,3-thiazol-2-yl) benzamide (as shown in the reaction formula (A-1) below) CAS Registry Number: 12817772-26-6), 5-amino-N- (4-t-butyl-1,3-thiazol-2-yl) -2-chlorobenzamide (CAS Registry Number: 13008582-83-3) Or 5-amino-N- (4-t-butyl-1,3-thiazol-2-yl) -2-bromobenzamide (CAS Registry Number: 130654) -17-1) is a condensing agent such as 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI), wherein a carboxylic acid in which a reactive group other than a carboxyl group is protected by an appropriate protecting group After the condensation in the presence, it can be synthesized by deprotecting the protecting group in the resulting condensate. As the protecting group for the amino group on the side chain, a widely used product such as a t-butoxycarbonyl (Boc) group or a 9-fluorenylmethyloxycarbonyl (Fmoc) group can be used. The deprotection reaction can be performed by a conventional method. More specifically, in the following reaction formula (A-1), the compound of the formula (H-31) can be synthesized by using D-lysine in which the side chain amino group is protected as the carboxylic acid. A compound of formula (H-36) can be synthesized by using L-lysine protected with an amino group, and a compound of formula (H-32) can be synthesized by using L-arginine with the amino group of the side chain protected. The compound of the formula (H-37) can be synthesized by using L-ornithine in which the side chain amino group is protected.

Further, for example, among compounds represented by the general formula (I), a compound having a common structure in which X is a hydrogen atom and a side chain containing R ² and R ³ is D-lysine is represented by (R ) -3- (2,6-bis ((t-butoxycarbonyl) amino) hexanamido) benzoic acid as a common raw material, and after condensation with 1-adamantylamine, the protective group is deprotected. A compound of formula (H-47) can be synthesized, and after condensation with hexylamine, formula (H-50) can be synthesized by deprotecting the protecting group. In addition, (R) -3- (2,6-bis ((t-butoxycarbonyl) amino) hexanamide) benzoic acid is obtained by combining ethyl 3-aminobenzoate and D-lysine in which the side chain amino group is protected. After condensing with 2-chloro-1-methylpyridinium iodide, it can be synthesized by hydrolyzing the ethyl ester of the resulting condensed compound with lithium hydroxide.

  Even if the compound represented by general formula (I) is a physiologically acceptable salt, it can be used as an active ingredient of a multidrug efflux pump inhibitor like the free form compound. Since the compound represented by the general formula (I) has a basic amino group, examples of physiologically acceptable salts of the compound include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphorus Salts of mineral acids such as acids; methanesulfonic acid, benzenesulfonic acid, paratoluenesulfonic acid, acetic acid, propionate, tartaric acid, fumaric acid, maleic acid, malic acid, oxalic acid, succinic acid, citric acid, benzoic acid, And salts with organic acids such as mandelic acid, cinnamic acid, lactic acid, glycolic acid, glucuronic acid, ascorbic acid, nicotinic acid and salicylic acid; or salts with acidic amino acids such as aspartic acid and glutamic acid.

  The compound represented by the general formula (I) can be used as an active ingredient of a multidrug efflux pump inhibitor even in the solvate of the compound or a physiologically acceptable salt thereof, like the free form compound. . Examples of the solvent that forms the solvate include water and ethanol.

  The multidrug discharge pump inhibitor according to the present invention may contain only one type of compound represented by the general formula (I) or may contain two or more types in combination. In addition, the compound represented by the general formula (I) contained in the multidrug efflux pump inhibitor according to the present invention may be only in a free form, and a free form compound and a physiologically acceptable salt thereof Both free form compounds and solvates thereof, both physiologically acceptable salts and solvates thereof, free form compounds and The physiologically acceptable salts and their solvates may be all.

  The multidrug efflux pump inhibitor according to the present invention is a compound represented by the general formula (I) which is an active ingredient or a physiologically acceptable salt thereof, or a solvate thereof (hereinafter referred to as “the present invention”). It may be composed only of “compounds and the like.”) And may contain various additives as long as the multidrug efflux pump inhibitory activity of the compound according to the present invention is not impaired. Such additives include excipients, binders, lubricants, wetting agents, solvents, disintegrants, solubilizers, suspending agents, emulsifiers, isotonic agents, stabilizers, buffers, preservatives. , Antioxidants, flavoring agents, coloring agents and the like. These additives are pharmaceutically acceptable substances and can be appropriately selected from those used for pharmaceutical preparation.

  The compound according to the present invention is also suitable as an active ingredient of a pharmaceutical composition. The pharmaceutical composition according to the present invention may contain only one type of the compound according to the present invention, or may contain two or more types in combination.

  Since the compound according to the present invention has the ability to inhibit the drug excretion ability of the RND type multidrug excretion pump possessed by Gram-negative bacteria, when used in combination with one or more antibacterial agents, The sensitivity to antibacterial agents can be increased and the resulting therapeutic effect can be enhanced. That is, by administering together with the compound etc. which concern on this invention, the dosage of an antibacterial agent required in order to acquire a required therapeutic effect can be decreased rather than the case where it administers alone. By reducing the dose of the antibacterial agent, the compound according to the present invention can also suppress the resistance of the bacterium to the antibacterial agent. Furthermore, the compound etc. which concern on this invention can act on the bacteria which acquired tolerance with respect to an antibacterial agent, can inhibit the discharge | emission ability of a multidrug discharge pump, and can also make it detolerated.

  Therefore, the pharmaceutical composition comprising the compound according to the present invention as an active ingredient is preferably used for the prevention and / or treatment of bacterial infections, and the bacteria that have acquired resistance to at least one antibacterial agent are pathogenic. What is used for prevention and / or treatment of bacterial infection which is bacteria is more preferable, and what is used for prevention and / or treatment of bacterial infection whose multidrug resistant bacteria are pathogenic bacteria is more preferable.

  The kind of bacterial infection which becomes the application target of the pharmaceutical composition according to the present invention is not particularly limited. The application target of the pharmaceutical composition according to the present invention is preferably a gram-negative bacterial infection (bacterial infection in which the pathogenic bacterium is a gram-negative bacterium), and a bacterium in which Pseudomonas aeruginosa, Acinetobacter, or enterococci are pathogenic bacteria. Infectious diseases are more preferred, bacterial infections in which Pseudomonas aeruginosa is a pathogenic bacterium are further preferred, and multidrug resistant Pseudomonas aeruginosa is particularly preferred.

  The pharmaceutical composition according to the present invention is preferably administered to humans or non-human animals. Examples of animals other than humans include mammals such as cows, pigs, horses, sheep, goats, monkeys, dogs, cats, rabbits, mice, rats, hamsters, guinea pigs, and birds such as chickens, quails, and ducks. Can be mentioned.

  As antibacterial agents, compounds having various structures have been known so far, and many compounds have been clinically used as pharmaceuticals. The kind of antibacterial agent that can be administered together with the pharmaceutical composition according to the present invention is not particularly limited. Examples of the antibacterial agent include penicillin (penam), cephalosporin (cephem), oxacephem, penem, carbapenem and other bicyclic β-lactam antibacterial agents, monobactams and the like. Cyclic β-lactam antibacterial agent, aminoglycoside antibacterial agent, macrolide antibacterial agent, chloramphenicol antibacterial agent, tetracycline antibacterial agent, glycopeptide antibacterial agent, fosfomycin antibacterial agent And lincomycin-based antibacterial agents, as well as sulfa drugs, paraaminosalicylic acid formulations, isonicotinic acid hydrazide formulations, and quinolone-based synthetic antibacterial agents.

  The pharmaceutical composition according to the present invention may contain other active ingredients in addition to the compound according to the present invention. When the pharmaceutical composition according to the present invention is a composition used for the prevention and / or treatment of bacterial infection, the active ingredient preferably contained in the pharmaceutical composition according to the present invention includes, for example, Examples thereof include agents having a function of suppressing resistance to antibacterial agents such as bacterial agents and β-lactamase inhibitors. When the pharmaceutical composition according to the present invention further contains one or more antibacterial agents, the antibacterial agent is listed as an antibacterial agent that can be administered together with the pharmaceutical composition according to the present invention. Can be mentioned. By administering a compound according to the present invention in combination with an antibacterial agent, an extremely high therapeutic effect can be achieved for bacterial infections.

  The pharmaceutical composition according to the present invention is known in the field of pharmaceutics by mixing the compound according to the present invention, which is an active ingredient, with one or more pharmaceutically acceptable excipients. According to the formulation method, it can be formulated into a dosage form suitable for administration to animals. Examples of pharmaceutical additives include excipients, binders, lubricants, wetting agents, solvents, disintegrants, solubilizers, suspending agents, emulsifiers (surfactants), isotonic agents, and stability. Examples include, but are not limited to, agents, buffers, plasticizers, preservatives, antioxidants, flavoring agents, and coloring agents.

  The method for administering the pharmaceutical composition according to the present invention to an animal is not particularly limited, and the dosage form is not particularly limited. Examples of dosage forms suitable for oral administration include capsules, powders, tablets, granules, fine granules, emulsions, syrups, solutions, suspensions, etc., and dosage forms suitable for parenteral administration As, for example, inhalants, sprays, rectal administration, injections, drops, ointments, creams, transdermal absorption agents, transmucosal absorption agents, eye drops, nasal drops, ear drops, tapes And patches.

  Among pharmaceutical compositions suitable for oral administration, for example, liquid preparations such as emulsions and syrups include water; sugars such as sucrose, sorbit and fructose; glycols such as polyethylene glycol and propylene glycol; sesame oil, olive oil, soybean oil and the like Oils; Preservatives such as p-hydroxybenzoates; and other additives for pharmaceutical use such as strawberry flavors and flavors such as peppermint. Solid preparations such as capsules, tablets, powders and granules include excipients such as lactose, glucose, sucrose and mannitol; disintegrants such as starch and sodium alginate; lubricants such as magnesium stearate and talc; It can be produced using a binder such as polyvinyl alcohol, hydroxypropyl cellulose, gelatin; a surfactant such as a fatty acid ester; a plasticizer such as glycerin.

  Among pharmaceutical compositions suitable for parenteral administration, liquid preparations such as injections, drops, eye drops and the like can be prepared preferably as sterilized isotonic liquid preparations. For example, injections can be prepared using an aqueous medium consisting of a salt solution, a glucose solution, or a mixture of saline and glucose solution. Rectal administration agents can be prepared usually in the form of suppositories, using a carrier such as cacao butter, hydrogenated fat or hydrogenated carboxylic acid. For the preparation of the propellant, a non-irritating carrier that facilitates absorption by dispersing the above-mentioned substance as an active ingredient as fine particles can be used. Examples of such a carrier include lactose and glycerin, and the form of the preparation can be selected from forms such as aerosol and dry powder.

  The dosage and number of administrations of the pharmaceutical composition according to the present invention are not particularly limited, and the type and severity of the bacterial infection, the subject of administration, so that the action effect of the compound according to the present invention is sufficiently exhibited. It can be set as appropriate according to the species, sex, age, weight, presence or absence of the underlying disease, mode of administration, and the like.

  EXAMPLES Next, although an Example etc. are shown and this invention is demonstrated further in detail, this invention is not limited to a following example etc.

[Example 1]
(R) -N- (4- (t-butyl) thiazol-2-yl) -2-chloro-5- (2,6-diaminohexanamide) benzamide (a compound represented by the formula (H-31); Hereinafter, it may be referred to as “H-31 compound”) was synthesized as follows.

(1) In a 30 mL eggplant flask, Nα, Nε-bis (t-butoxycarbonyl) -D-lysine (215 mg, 0.62 mmol, 1.3 eq) was dissolved in dichloromethane, and 1- (3-dimethylamino was dissolved in dichloromethane. Propyl) -3-ethylcarbodiimide hydrochloride (EDCI; 151 mg, 0.79 mmol, 1.6 eq) was added dropwise. Next, commercially available 5-amino-N- (4-tert-butyl-1,3-thiazol-2-yl) -2-chlorobenzamide (152 mg, 0.49 mmol) dissolved in dichloromethane was added, and 3 Stir for hours. The reaction solution was washed successively with 1M aqueous hydrochloric acid solution, aqueous sodium hydrogen carbonate solution and saturated brine, then extracted with a mixed solvent of ethyl acetate and diethyl ether, and dried over sodium sulfate. The extract solution after drying was concentrated under reduced pressure, and silica gel column chromatography (hexane: ethyl acetate = 2: 1) was performed twice to obtain a condensate (200 mg, 0.31 mmol). The yield was 64%. NMR analysis and mass spectrometry were performed on the obtained condensate.

¹ H-NMR (400 MHz, CDCl ₃ ): δ 11.29 (br, 1H), 9.39 (brs, 1H), 7.67 (d, J = 8.7 Hz, 1H), 7.18 (brm, 1H), 7.12 (d, J = 8.7 Hz, 1H), 6.94 (brs, 1H), 6.61 (s, 1H), 4.55 (brm, 1H), 4.47 (q , J = 7.8 Hz, 1H), 3.11 (brm, 2H), 2.00-1.80 (m, 2H), 1.62-1.46 (m, 4H), 1.40 (s , 9H), 1.37 (s, 9H), 1.25 (s, 9H).
HRMS (ESI, m / z, M + Na ⁺ ): Calculated 660.2593 for C ₃₀ H ₄₄ ClN ₅ O ₆ SNa, found 660.2596.

(2) Next, the condensate (63 mg, 0.10 mmol) obtained in the above step (1) was dissolved in diethyl ether (3 mL) in a 20 mL eggplant flask, and then 1M aqueous hydrochloric acid / ethyl acetate (3 mL) was added. The solution was added dropwise and stirred at room temperature for 1 hour. The precipitated solid was subjected to suction filtration, and the collected solid was washed with diethyl ether and then vacuum-dried to obtain the target compound (H-31 compound) as a dihydrochloride salt. The yield was 51 mg (0.10 mmol), and the yield was 100%. The obtained target compound was subjected to NMR analysis and mass spectrometry.

¹ H-NMR (400 MHz, CD ₃ OD): δ 8.17 (d, J = 2.7 Hz, 1H), 7.88 (dd, J = 8.7, 2.7 Hz, 1H), 7.57 ( d, J = 8.7 Hz, 1H), 7.11 (s, 1H), 4.16 (brt, J = 6.6 Hz, 1H), 2.97 (brt, J = 7.8 Hz, 2H), 2.08 (m, 1H), 2.01 (m, 1H), 1.75 (m, 2H), 1.60 (m, 2H), 1.41 (s, 9H).
_{^{HRMS (ESI, m / z,}} M + H +): C 20 H 29 ClN 5 O 2 Calcd 438.1725 for S, Found 438.1717.

[Example 2]
(S) -N- (4- (t-butyl) thiazol-2-yl) -2-chloro-5- (2,6-diaminohexaneamide) benzamide (a compound represented by the formula (H-36); Hereinafter, it may be referred to as “H-36 compound”) was synthesized as follows.

(1) Nα, Nε-bis (t-butoxycarbonyl) -L-lysine is used instead of Nα, Nε-bis (t-butoxycarbonyl) -D-lysine, and the same as step (1) of Example 1 By the process, a condensate was obtained and subjected to NMR analysis and mass spectrometry. The yield was 92%.

¹ H-NMR (400 MHz, CDCl ₃ ): δ 11.38 (br, 1H), 9.41 (br, 1H), 7.66 (d, J = 8.7 Hz, 1H), 7.26 (br, 1H), 7.11 (d, J = 8.7 Hz, 1H), 6.91 (s, 1H), 6.61 (s, 1H), 4.57 (m, 1H), 4.49 (q , J = 7.8 Hz, 1H), 3.17-3.05 (br, 2H), 2.00-1.80 (m, 2H), 1.62-1.46 (m, 4H), 1 .40 (s, 9H), 1.36 (s, 9H), 1.24 (s, 9H).
HRMS (ESI, m / z, M + Na ⁺ ): Calculated 660.2593 for C ₃₀ H ₄₄ ClN ₅ O ₆ SNa, found 660.25592.

(2) Next, the target compound (H-36 compound) was obtained as a dihydrochloride salt from the condensate obtained in the step (1) by the same step as the step (2) of Example 1, and subjected to NMR analysis. And mass spectrometry was performed. The yield was 100%.

¹ H-NMR (400 MHz, CD ₃ OD): δ 8.12 (brm, 1H), 7.86 (brd, J = 8.7 Hz, 1H), 7.56 (d, J = 8.7 Hz, 1H) 7.02 (brs, 1H), 4.15 (m, 1H), 2.97 (brt, J = 7.8 Hz, 2H), 2.08 (m, 1H), 2.01 (m, 1H) ), 1.75 (m, 2H), 1.60 (m, 2H), 1.39 (s, 9H).
HRMS (ESI, m / z, M + H ⁺ ): Calculated 438.1725 for C ₂₀ H ₂₉ ClN ₅ O ₂ S, found 438.1726.

[Example 3]
(R) -N- (4- (t-butyl) thiazol-2-yl) -2-bromo-5- (2,6-diaminohexanamide) benzamide (a compound represented by the formula (H-34); Hereinafter, it may be referred to as “H-34 compound”) was synthesized as follows.

(1) From commercially available 5-amino-N- (4-t-butyl-1,3-thiazol-2-yl) -2-bromobenzamide and Nα, Nε-bis (t-butoxycarbonyl) -D-lysine The condensate was obtained by the same process as the process (1) of Example 1, and NMR analysis and mass spectrometry were performed. The yield was 81%.

¹ H-NMR (400 MHz, CDCl ₃ ): δ 11.27 (br, 1H), 9.39 (brs, 1H), 7.57 (d, J = 8.7 Hz, 1H), 7.29 (d, J = 8.7 Hz, 1H), 7.25 (brm, 1H), 6.89 (brs, 1H), 6.61 (s, 1H), 4.55 (brm, 1H), 4.47 (q , J = 7.8 Hz, 1H), 3.12 (brm, 2H), 1.98-1.78 (m, 2H), 1.67-1.42 (m, 4H), 1.40 (s , 9H), 1.36 (s, 9H), 1.24 (s, 9H).
HRMS (ESI, m / z, M + Na ⁺ ): calcd for C ₃₀ H ₄₄ BrN ₅ O ₆ SNa 704.2088, found 704.2095.

(2) Next, the target compound (H-34 compound) was obtained as a dihydrochloride salt from the condensate obtained in the step (1) by the same step as the step (2) of Example 1, and subjected to NMR analysis. And mass spectrometry was performed. The yield was 100%.

¹ H-NMR (400 MHz, CD ₃ OD): δ 8.04 (d, J = 2.2 Hz, 1H), 7.76 (dd, J = 8.6, 2.2 Hz, 1H), 7.71 ( d, J = 8.7 Hz, 1H), 6.95 (s, 1H), 4.15 (brt, J = 6.2 Hz, 1H) 2.97 (brt, J = 7.5 Hz, 2H), 2 .06 (m, 1H), 1.98 (m, 1H), 1.75 (m, 2H), 1.57 (m, 2H), 1.37 (s, 9H).
HRMS (ESI, m / z, M + H ⁺ ): Calculated 482.1220 for C ₂₀ H ₂₉ BrN ₅ O ₂ S, found 482.1219.

[Example 4]
(S) -N- (4- (t-butyl) thiazol-2-yl) -2-chloro-5- (2,5-diaminopentanamide) benzamide (a compound represented by the formula (H-37); Hereinafter, it may be referred to as “H-37 compound”) was synthesized as follows.

(1) In place of Nα, Nε-bis (t-butoxycarbonyl) -D-lysine, Nα, Nδ-bis (t-butoxycarbonyl) -L-ornithine is used, and the same as step (1) of Example 1 By the process, a condensate was obtained and subjected to NMR analysis and mass spectrometry. The yield was 29%.

¹ H-NMR (400 MHz, CDCl ₃ ): δ 11.15 (br, 1H), 9.39 (brs, 1H), 7.65 (brd, J = 8.7 Hz, 1H), 7.13 (d, J = 8.7 Hz, 1H), 7.03 (brm, 2H), 6.61 (s, 1H), 4.69 (brm, 1H), 4.53 (brq, J = 7.3 Hz, 1H) 3.28 (m, 1H), 3.18 (m, 1H), 1.93 (m, 1H), 1.84 (m, 1H), 1.73-1.60 (m, 2H), 1.40 (s, 9H), 1.36 (s, 9H), 1.26 (s, 9H).
HRMS (ESI, m / z, M + Na ⁺ ): calcd for C ₂₉ H ₄₂ ClN ₅ O ₆ SNa 646.2437, found 646.2436.

(2) Next, the target compound (H-37 compound) was obtained as a dihydrochloride salt from the condensate obtained in the step (1) by the same step as the step (2) of Example 1, and subjected to NMR analysis. And mass spectrometry was performed. The yield was 70%.

¹ H-NMR (400 MHz, CD ₃ OD): δ 8.12 (d, J = 2.3 Hz, 1H), 7.86 (dd, J = 8.7, 2.3 Hz, 1H), 7.56 ( d, J = 8.7 Hz, 1H), 7.01 (s, 1H), 4.20 (brt, J = 6.6 Hz, 1H), 3.11-2.93 (m, 2H), 2. 19-1.95 (m, 2H), 1.95-1.78 (m, 2H), 1.38 (s, 9H).
HRMS (ESI, m / z, M + H ⁺ ): Calculated 414.1568 for C ₁₉ H ₂₇ ClN ₅ O ₂ S, found 424.1569.

[Example 5]
(S) -5- (2-amino-5-guanidinopentanamide) -N- (4- (t-butyl) thiazol-2-yl) -2-chlorobenzamide (represented by formula (H-32) Compound, hereinafter sometimes referred to as “H-32 compound”) was synthesized as follows.

(1) N-α- (9-fluorenylmethoxycarbonyl) -N-ω- (2,2,4,6,7- in place of Nα, Nε-bis (t-butoxycarbonyl) -D-lysine Using pentamethyldihydrobenzofuran-5-sulfonyl) -L-arginine, a condensate was obtained in the same manner as in step (1) of Example 1, and NMR analysis and mass spectrometry were performed. The yield was 58%.

¹ H-NMR (400 MHz, CD ₃ OD): δ 7.78 (d, J = 2.3 Hz, 1H), 7.68 (d, J = 7.8 Hz, 2H), 7.60 (dd, J = 8.7, 2.3 Hz, 1H), 7.56 (dd in appearance, J = 6.8, 5.5 Hz, 2H), 7.37 (d, J = 8.7 Hz, 1H), 7.26. (Dd, J = 7.8, 7.3 Hz, 2H), 7.19 (dd, J = 7.8, 7.3 Hz, 2H), 6.66 (s, 1H), 4.53 (brs, 1H), 4.31 (d, J = 6.9 Hz, 2H), 4.12 (m, 2H), 3.15 (m, 1H), 2.85 (s, 2H), 2.46 (s) , 3H), 2.39 (s, 3H), 1.94 (s, 3H), 1.72 (m, 1H), 1.60 (m, 1H), 1.54-1.1.13 ( m, 2H), 1.31 (s, 6H), 1.22 (s, 9H).
_{^{HRMS (ESI, m / z,}} M + Na +): C 48 H 54 ClN 7 O 7 S Calculated 962.3107 for 2 Na, Found 962.3287.

(2) Next, 0.5 mL of piperidine was added to a dichloromethane solution (2 mL) of the condensate (127 mg, 0.135 mmol) obtained in the step (1), and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was diluted with ethyl acetate, the organic phase was extracted three times with 1M aqueous hydrochloric acid solution to remove neutral / acidic impurities, and the aqueous phase was neutralized with aqueous sodium hydrogen carbonate solution. The neutralized solution was extracted with a mixed solvent of ethyl acetate and diethyl ether, and the obtained organic phase was washed with saturated brine and then dried over sodium sulfate. Next, after concentrating the organic phase, purification was performed with silica gel column chromatography (dichloromethane: methanol = 9: 1) to obtain 83 mg (0.12 mmol) of Fmoc deprotected body, and NMR analysis and mass spectrometry were performed. It was 86%.

¹ H-NMR (400 MHz, CD ₃ OD): δ 7.82 (d, J = 2.8 Hz, 1H), 7.60 (dd, J = 8.7, 2.8 Hz, 1H), 7.36 ( d, 8.7 Hz, 1H), 6.65 (s, 1H) 3.36 (dd, J = 6.4, 5.5 Hz, 1H), 3.28-3.02 (m, 2H), 2 .87 (s, 2H), 2.45 (s, 3H), 2.38 (s, 3H), 1.95 (s, 3H), 1.71-1.42 (m, 4H), 1. 33 (s, 6H), 1.22 (s, 9H).
_{^{HRMS (ESI, m / z,}} M + H +): C 33 H 45 ClN 7 O 5 Calculated 718.2607 for _{S 2,} Found 718.2607.

(3) Next, the Fmoc deprotected body (53 mg, 0.074 mmol) obtained in the above step (2) was dissolved in 2 mL of trifluoroacetic acid, 0.3 mL of water was added, and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was diluted with water and extracted with diethyl ether. Specifically, after the aqueous phase was evaporated to dryness, diethyl ether was added to the obtained solid, and insoluble matters were removed by filtration to obtain a diethyl ether solution of the target compound (H-32 compound). . On the other hand, the basic compound was re-extracted from the diethyl ether phase with 1M hydrochloric acid, and the resulting extract was neutralized with an aqueous sodium hydrogen carbonate solution and then extracted with ethyl acetate. The obtained organic phase was washed with saturated brine, dehydrated with sodium sulfate, concentrated with the diethyl ether solution obtained first, and further dried in vacuo overnight to give the desired compound (white powder solid) H-32 compound) (23 mg, 0.049 mmol) was obtained. The yield was 66%.

¹ H-NMR (400 MHz, CD ₃ OD): δ 7.91 (d, J = 2.8 Hz, 1H), 7.74 (dd, J = 9.2, 2.8 Hz, 1H), 7.42 ( d, J = 9.2 Hz, 1H), 6.79 (s, 1H), 4.02 (brdd, J = 6.0, 5.5 Hz, 1H), 3.17 (brdd, J = 6.9). , 6.4 Hz, 2H), 1.98 (m, 1H), 1.91 (m, 1H), 1.67 (m, 2H), 1.26 (s, 9H).
_{^{HRMS (ESI, m / z,}} M + H +): C 20 H 29 ClN 7 O 2 Calcd 466.1786 for S, Found 466.1777.

[Example 6]
5- (2-aminohexanamide) -N- (4- (t-butyl) thiazol-2-yl) -2-chlorobenzamide (compound represented by formula (H-35), hereinafter referred to as “H-35 compound” Was synthesized as follows.

(1) A step similar to step (1) of Example 1 except that 6-((t-butoxycarbonyl) amino) hexanoic acid is used instead of Nα, Nε-bis (t-butoxycarbonyl) -D-lysine. The condensate was obtained by NMR analysis. The yield was 89%.

¹ H-NMR (400 MHz, CDCl ₃ ): δ 9.91 (br, 1H), 8.26 (br, 1H), 7.91 (s, 1H), 7.83 (d, J = 8.7 Hz, 1H), 7.38 (d, J = 8.7 Hz, 1H), 6.60 (s, 1H), 4.68 (brm, 1H), 3.10 (m, 2H), 2.38 (m , 2H), 1.71 (m, 2H), 1.48 (m, 2H), 1.43 (s, 9H), 1.35 (m, 2H), 1.30 (s, 9H).

(2) Next, from the condensate obtained in the step (1), the target compound (H-35 compound) is obtained as a hydrochloride by the same step as the step (2) of Example 1, and NMR analysis is performed. went. The yield was 85%.

¹ H-NMR (400 MHz, CD ₃ OD): δ 7.96 (d, J = 2.6 Hz, 1H), 7.70 (dd, J = 8.6, 2.6 Hz, 1H), 7.49 ( d, J = 8.6 Hz, 1H), 6.88 (s, 1H), 2.93 (brdd, J = 7.7, 7.3 Hz, 2H), 2.45 (t, J = 7.3 Hz). , 2H), 1.75 (ttm, J = 7.7, 7.3 Hz, 2H), 1.70 (ttm, J = 7.7, 7.3 Hz, 2H), 1.47 (m, 2H) , 1.35 (s, 9H).

[Example 7]
N- (4- (t-butyl) thiazol-2-yl) -2-chloro-5- (2,3-diaminopropanamide) benzamide (a compound represented by the formula (H-39), hereinafter referred to as “H- 39 compounds ") was synthesized as follows.

(1) In place of Nα, Nε-bis (t-butoxycarbonyl) -D-lysine, 2,3-bis ((t-butoxycarbonyl) amino) propanoic acid was used, and step (1) of Example 1 and A condensate was obtained by the same process and subjected to NMR analysis. The yield was 87%.

¹ H-NMR (400 MHz, CDCl ₃ ): δ 10.46 (br, 1H), 9.43 (brs, 1H), 7.69 (s, 1H), 7.66 (d, J = 8.4 Hz, 1H), 7.31 (d, J = 8.4 Hz, 1H), 6.61 (d, J = 4.9 Hz, 1H), 6.35 (brs, 1H), 5.38 (brm, 1H) 4.42 (brm, 1H), 3.63 (m, 1H), 3.56 (m, 1H), 1.41 (s, 9H), 1.40 (s, 9H), 1.33 ( s, 9H).

(2) Next, the target compound (H-39 compound) was obtained as a dihydrochloride salt from the condensate obtained in the step (1) by the same step as the step (2) of Example 1, and subjected to NMR analysis. Went. The yield was 100%.

¹ H-NMR (700 MHz, CD ₃ OD): δ 8.10 (d, J = 2.6 Hz, 1H), 7.86 (dd, J = 8.8, 2.6 Hz, 1H), 7.56 ( d, J = 8.8 Hz, 1H), 6.90 (s, 1H), 4.56 (dd, J = 5.8, 5.6 Hz, 1H), 3.62 (dd, J = 14.1) , 5.8 Hz, 1H), 3.54 (dd, J = 14.1, 5.6 Hz, 1H), 1.35 (s, 9H).

[Example 8]
(S) -5- (2-Amino-3- (1H-imidazol-4-yl) propanamide-N- (4- (t-butyl) thiazol-2-yl) -2-chlorobenzamide (formula (H The compound represented by -46), hereinafter sometimes referred to as "H-46 compound") was synthesized as follows.

(1) Instead of Nα, Nε-bis (t-butoxycarbonyl) -D-lysine, N-α- (9-fluorenylmethoxycarbonyl) amino) -N-τ-trityl-L-histidine was used. A condensate was obtained by the same step as in step (1) of Example 1, and NMR analysis was performed. The yield was 37%.

¹ H-NMR (400 MHz, CDCl ₃ ): δ 11.01 (br, 1H), 10.01 (brs, 1H), 7.66-7.61 (m, 3H), 7.45-7.10 ( m, 19H), 6.97 (d-like, J = 7.8 Hz, 6H), 6.63 (s, 1H), 6.58 (s, 1H), 4.84 (dm, J = 6. 4 Hz, 1 H), 4.33 (dd, J = 11.3, 7.1 Hz, 1 H), 4.16 (m, 1 H), 3.99 (brt, J = 6.9 Hz, 1 H), 3. 37 (br, 1H), 3.15-3.04 (m, 2H), 1.29 (s, 9H).

(2) Next, from the condensate obtained in the step (1), an Fmoc deprotection product was obtained in the same manner as the step (2) of Example 5, and NMR analysis was performed. The yield was 42%.

¹ H-NMR (400 MHz, CDCl ₃ ): δ 10.06 (br, 1H), 7.83 (s, 1H), 7.63 (d, J = 8.2 Hz, 1H), 7.30-7. 20 (m, 11H), 7.03 (brd, J = 6.9 Hz, 6H), 6.61 (s, 1H), 6.57 (s, 1H), 3.77 (brm, 1H), 3 .02 (m, 1H), 2.91 (m, 1H), 1.26 (s, 9H).

(3) Next, the target compound (H-46 compound) was obtained from the Fmoc deprotected body obtained in the step (2) by the same step as the step (3) of Example 5, and subjected to NMR analysis. It was. The yield was 52%.

¹ H-NMR (400 MHz, CDCl ₃ ): δ 9.94 (br, 1H), 7.95 (d, J = 2.3 Hz, 1H), 7.74 (d, J = 8.7, 2.3 Hz) , 1H), 7.54 (s, 1H), 7.38 (d, J = 8.7 Hz, 1H), 6.85 (s, 1H), 6.61 (s, 1H), 3.77 ( t, J = 5.5 Hz, 1H), 3.11 (dd, J = 14.7, 5.5 Hz, 1H), 3.06 (dd, J = 14.7, 5.5 Hz, 1H), 1 .30 (s, 9H).

[Production Example 1]
(R) -3- (2,6-bis ((t-butoxycarbonyl) amino) hexanamido) benzoic acid was synthesized as follows.

(1) Ethyl 3-aminobenzoate (366 mg, 2.22 mmol) and Nα, Nε-bis (t-butoxycarbonyl) -D-lysine (969 mg, 2.93 mmol) were dissolved in dichloromethane (8 mL), and 2- Chloro-1-methylpyridinium iodide (879 mg, 3.44 mmol) and triethylamine (1.5 mL, 10 mmol) were added, and the mixture was stirred at room temperature for 12 hours. After completion of the reaction, the reaction mixture was diluted with ethyl acetate, and the organic phase was washed with water and then with an aqueous sodium hydrogen sulfate solution. The aqueous phase at that time was re-extracted twice with ethyl acetate, and the organic phase was washed successively with an aqueous sodium hydrogen carbonate solution and saturated brine, and then dried over sodium sulfate. The extracted solution after drying was concentrated under reduced pressure and purified by silica gel column chromatography (hexane: ethyl acetate = 2: 1) to obtain a condensate (456 mg, 0.924 mmol). The yield was 42%. NMR analysis was performed on the resulting condensate.

¹ H-NMR (400 MHz, CDCl ₃ ): δ 9.16 (br, 1H), 8.10 (s, 1H), 7.80-7.68 (m, 2H), 7.30 (brm, 1H) , 5.61 (brs, 1H), 4.80 (brs, 1H), 4.38-4.32 (m, 3H), 3.18-3.06 (m, 2H), 1.91 (m , 1H), 1.74 (m, 1H), 1.58-1.40 (m, 4H), 1.44 (s, 9H), 1.43 (s, 9H), 1.37 (t, J = 7.0 Hz, 3H).

(2) Next, the condensate (972 mg, 1.97 mmol) obtained in the step (1) was dissolved in tetrahydrofuran (20 mL), 10 mL each of water and methanol was added, and lithium hydroxide monohydrate was further added. (234 mg, 5.58 mmol) was added, and the mixture was stirred at room temperature for 2.5 hours. After completion of the reaction, the mixture was diluted with an aqueous potassium carbonate solution and washed with ethyl acetate to remove neutral and basic substances. The aqueous phase was acidified with 1M hydrochloric acid dropwise, and extracted with ethyl acetate. The organic phase was washed with saturated brine and dried over sodium sulfate. The extracted solution after drying was concentrated under reduced pressure to obtain the desired benzoic acid derivative (864 mg, 1.86 mmol). The yield was 96%. The obtained target compound was subjected to NMR analysis.

¹ H-NMR (400 MHz, CDCl ₃ ): δ 9.50 (s, 1H), 8.23 (brs, 1H), 8.14 (s, 1H), 7.79 (d, J = 6.9 Hz, 1H), 7.38 (dd, J = 7.6, 6.9 Hz, 1H), 5.81 (brs, 1H), 4.81 (brs, 1H), 4.46 (brs, 1H), 3 .10 (brm, 2H), 1.87 (m, 1H), 1.75 (m, 1H), 1.55-1.50 (m, 2H), 1.46 (s, 9H), 1. 41 (s, 9H).

[Example 9]
(R) -N-adamantan-1-yl-3- (2,6-diaminohexanamide) benzamide (compound represented by the formula (H-47), hereinafter may be referred to as “H-47 compound”) Was synthesized as follows.

(1) 1-adamantylamine (55 mg, 0.36 mmol) and the benzoic acid derivative (147 mg, 0.316 mmol) obtained in Preparation Example 1 were dissolved in dichloromethane (8 mL), and EDCI hydrochloride (64 mg, 0.335 mmol) was dissolved. ), Diisopropylethylamine (610 μL, 3.59 mmol) and 1-hydroxybenzotriazole (HOBt: 45 mg, 0.33 mmol) were sequentially added, and the mixture was stirred at room temperature for 4 hours. After completion of the reaction, the reaction mixture was diluted with ethyl acetate, and the organic phase was washed with water and then with an aqueous sodium hydrogen sulfate solution. Further, the organic phase was washed successively with an aqueous sodium hydrogen carbonate solution and saturated brine, and then dried over sodium sulfate. The extracted solution after drying was concentrated under reduced pressure and purified by silica gel column chromatography (hexane: ethyl acetate = 2: 1) to obtain a condensate (115 mg, 0.192 mmol). The yield was 60%. NMR analysis was performed on the resulting condensate.

¹ H-NMR (400 MHz, CDCl ₃ ): δ 9.18 (brs, 1H), 7.64 (d, J = 7.8 Hz, 1H), 7.40 (brs, 1H), 7.36 (d, J = 7.3 Hz, 1H), 7.15 (dd, J = 7.8, 7.3 Hz, 1H), 6.31 (s, 1H), 5.44 (brm, 1H), 4.70 ( brm, 1H), 4.31 (brm, 1H), 3.11 (brm, 2H), 2.16 (brs, 4H), 2.14 (brm, 4H), 1.99 (m, 2H), 1.95-1.81 (m, 2H), 1.80-1.48 (m, 9H), 1.47 (s, 9H), 1.44 (s, 9H).

(2) Next, the condensate (109 mg, 0.182 mmol) obtained in the above step (1) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added dropwise thereto at room temperature for 1.5 hours. Stir. After completion of the reaction, the reaction solution is concentrated, and the solid precipitated by adding diethyl ether is filtered by suction. The collected solid is washed with diethyl ether and further dried under vacuum to obtain the target compound (H-47 compound) as 2 Obtained as the trifluoroacetate salt. The yield was 85 mg (0.213 mmol), and the yield was 75%. The obtained target compound was subjected to NMR analysis.

¹ H-NMR (400 MHz, CD ₃ OD): δ 7.98 (s, 1H), 7.72 (dm, J = 8.2 Hz, 1H), 7.49 (d, J = 7.8 Hz, 1H) , 7.40 (dd, J = 8.2, 7.8 Hz, 1H), 4.06 (brt, J = 6.4 Hz, 1H), 2.94 (dd, J = 7.8, 7.3 Hz) , 2H), 2.15 (m, 6H), 2.10 (brm, 3H), 2.06-1.92 (m, 2H), 1.80-1.68 (m, 8H), 1. 60-1.49 (m, 2H).

[Example 10]
(R) -3- (2,6-diaminohexanamide) -N-hexylbenzamide (a compound represented by the formula (H-50), hereinafter may be referred to as “H-50 compound”) is as follows. Synthesized as street.

(1) Using hexylamine instead of 1-adamantylamine, a condensate was obtained and subjected to NMR analysis by the same step as in step (1) of Example 9. The yield was 45%.

¹ H-NMR (400 MHz, CDCl ₃ ): δ9.53 (brs, 1H), 7.68 (d, J = 7.8 Hz, 1H), 7.33 (d, J = 6.9 Hz, 1H), 7.14-7.05 (m, 2H), 5.58 (brm, 1H), 4.74 (brs, 1H), 4.37 (brm, 1H), 3.50-3.36 (m, 2H), 3.11 (brm, 2H), 1.86-1.28 (m, 15H), 1.44 (s, 9H), 1.43 (s, 9H), 0.91 (m, 3H) ).

(2) Next, the target compound (H-50 compound) was obtained as a 2-trifluoroacetate salt from the condensate obtained in the step (1) by the same step as the step (2) in Example 9. NMR analysis was performed. The yield was 72%.

¹ H-NMR (400 MHz, CD ₃ OD): δ 8.09 (s, 1H), 7.76 (d, J = 7.8 Hz, 1H), 7.56 (d, J = 7.8 Hz, 1H) , 7.43 (t, J = 7.8 Hz, 1H), 4.03 (t, J = 6.4 Hz, 1H), 3.37 (dd, J = 7.3, 6.9 Hz, 2H), 2.94 (brt, J = 7.8 Hz, 2H), 2.08-1.94 (m, 2H), 1.77-1.69 (m, 2H), 1.64-1.51 (m , 4H), 1.42-1.30 (m, 6H), 0.91 (t-like, J to 6.8 Hz, 3H).

[Example 11]
(S) -N- (4- (t-butyl) thiazol-2-yl) -3- (2,6-diaminohexaneamide) benzamide (compound represented by formula (H-51), hereinafter referred to as “H- 51 compounds ”)) was synthesized as follows.

(1) Instead of Nα, Nε-bis (t-butoxycarbonyl) -D-lysine, Nα, Nε-bis (t-butoxycarbonyl) -L-lysine is used, and 5-amino-N- (4-t Example using 3-amino-N- (4-tert-butyl-1,3-thiazol-2-yl) benzamide instead of -butyl-1,3-thiazol-2-yl) -2-chlorobenzamide A condensate was obtained by the same step as step 1 (1) and NMR analysis was performed. The yield was 90%.

¹ H-NMR (400 MHz, CDCl ₃ ): δ 10.84 (br, 1H), 9.35 (brs, 1H), 7.70 (d, J = 7.8 Hz, 1H), 7.54 (d, J = 7.3 Hz, 1H), 7.27 (brs, 1H), 7.11 (dd, J = 7.8, 7.3 Hz, 1H), 6.79 (d, J = 8.2 Hz, 1H) ), 6.58 (s, 1H), 4.68 (brm, 1H), 4.51 (dtm, J = 7.3, 6.9 Hz, 1H), 3.13 (brm, 2H), 1. 95 (m, 1H), 1.88 (m, 1H), 1.60-1.44 (m, 4H), 1.41 (s, 9H), 1.36 (s, 9H), 1.31 (S, 9H).

(2) Next, the target compound (H-51 compound) is obtained as a dihydrochloride salt from the condensate obtained in the step (1) by the same step as the step (2) in Example 1, and subjected to NMR analysis. Went. The yield was 100%.

¹ H-NMR (400 MHz, CD ₃ OD): δ 8.39 (dd, J = 2.3, 1.8 Hz, 1H), 7.92 (ddm, J = 8.2, 2.3 Hz, 1H), 7.86 (ddm, J = 8.2, 1.8 Hz, 1H), 7.58 (t, J = 8.2 Hz, 1H), 6.94 (s, 1H), 4.16 (t, J = 6.4 Hz, 1H), 2.98 (brdd, J = 7.8, 7.3 Hz, 2H), 2.14-1.97 (m, 2H), 1.81-1.73 (m, 2H), 1.68-1.53 (m, 2H), 1.39 (s, 9H).

[Test Example 1] Evaluation of the discharge inhibitory effect on the RND type multidrug discharge pump Erythromycin, which is a macrolide antibacterial agent and is an excretion substrate of MexB and MexY (hereinafter referred to as “EM” in the figures). The ability of the compound represented by the general formula (I) to inhibit the discharge ability of the compound represented by the general formula (I) on the RND type multidrug discharge pump was examined using the growth ability of bacteria in the presence of When the test compound has the ability to inhibit the discharge capacity of the RND-type multidrug discharge pump, the combined use of EM and the test compound suppresses bacterial growth compared to when EM is used alone.

Among the multidrug efflux system AcrAB-TolC possessed by E. coli described in Non-Patent Document 6, the Escherichia coli strain lacking the acrB and tolC genes is the main component of Pseudomonas aeruginosa described in Non-Patent Document 5. Mutant strain transfected with the gene of multidrug efflux system MexAB-OprM as a plasmid (hereinafter referred to as “pmexB strain” in the figure and the figure), another main multidrug efflux system gene of Pseudomonas aeruginosa MexXY-OprM The transferred strain (hereinafter referred to as “pmexY”) and the empty vector transferred strain (hereinafter referred to as “vector strain” in the figure) are represented by the compound represented by the general formula (I). Used to evaluate the inhibitory capacity against multidrug efflux pumps. For culture of each E. coli strain, LB (Luria-Bertani) broth was used as a medium, a 96-well plate (medium amount per well was 200 μL), and the amount of inoculum was 2 × 10 ⁷ CFU / μL. .

Specifically, each E. coli strain is cultured in a medium to which EM and various concentrations (0 to 128 μg / mL) of a test compound are added, and the growth of the strain is measured over time with optical turbidity (OD ₆₀₀ ) at ₆₀₀ nm. Was evaluated by mechanical measurement. The EM concentration of the medium was 8 μg / mL in the culture of the pmexB strain and 64 μg / mL in the culture of the pmexY strain.

  The contribution of the antibacterial activity of each test compound itself to the growth of the strain is to measure the growth of the strain when the same concentration of the test compound is added to the vector strain inoculated in the EM-free medium. Evaluated by.

OD _{600 of} the culture medium in which each E. coli strain was cultured when H-31 compound, H-32 compound, H-34 compound, H-36 compound, and H-37 compound were used as test compounds and each test compound was added. The measurement results are shown in FIGS. In FIGS. 1 to 5, (A) shows the results of the vector strain, (B) shows the results of the pmexB strain, and (C) shows the results of the pmexY strain. In the culture experiment of the vector strain, in any test compound, in the absence of EM, the OD ₆₀₀ decreased depending on the amount added to the medium, and the growth of the strain was suppressed (each figure (A)). From these results, it was found that these test compounds have weak antibacterial activity alone. In addition, when any of the test compounds was used in combination with EM, the OD ₆₀₀ was drastically decreased at an addition concentration of 8 μg / mL in both the pmexB strain and the pmexY strain, and the growth of each strain was suppressed (each figure ( B) and (C)). Since the antibacterial activity of each test compound was weak when the concentration added to the medium was 8 μg / mL, the decrease in the strain growth ability of the pmexB strain and the pmexY strain when combined with EM was due to the antibacterial activity of each test compound. It was found that each test compound was due to the antibacterial activity of EM accumulated in the strain by inhibiting the RND type multidrug efflux pump.

FIG. 6 shows the results of examining the growth of each strain in the same manner, with the H-31 compound as the test compound and the concentration added to the medium being lower (0 to 8 μg / mL). The H-31 compound showed almost no antibacterial activity at an addition concentration as low as 4 μg / mL (FIG. 6 (A)), but in both pmexB and pmexY strains, the addition was as low as 4 μg / mL. The OD ₆₀₀ was reduced to almost 0 with the concentration (FIGS. 6B and 6C). There is almost no difference in the OD ₆₀₀ of the medium when the addition concentration of H-31 compound is 2 μg / mL and when it is not added (0 μg / mL), and the growth ability is almost completely inhibited when the addition concentration is 4 μg / mL. From these results, it was suggested that there is a clear threshold for the ability of the compound of the present invention to inhibit the RND type multidrug efflux pump.

In addition to the H-35 compound, H-39 compound, H-46 compound, H-47 compound, H-50 compound, and H-51 compound synthesized in Examples 6 to 11, the following formula (H-8 ), (H-27) to (H-30), (H-33), (H-38), (H-41), (H-42), (H-45), (H-48), And about the compound represented by (H-49), the discharge ability inhibitory effect with respect to a RND type | mold multidrug discharge pump was investigated similarly. From the measurement result of OD ₆₀₀ of the culture medium in which each E. coli strain was cultured when each test compound was added, the growth inhibitory effect of each test compound was evaluated. The growth inhibitory effect of the test compound alone against the vector strain is evaluated as “−” when the EC ₅₀ (50% effective concentration) is 32 μg / mL or more in the experiment using the vector strain, and is 8 μg / mL or more. When it was less than 32 μg / mL, it was evaluated as “+”, and when it was less than 8 μg / mL, it was evaluated as “++”. The growth inhibitory effect of each test compound in combination with EM (8 μg / mL) against the pmexB strain was evaluated as “−” when the EC ₅₀ was 128 μg / mL or more in the experiment using the pmexB strain, and 8 μg When it was not less than 128 μg / mL, it was evaluated as “+”, and when it was less than 8 μg / mL, it was evaluated as “++”. The growth inhibitory effect of each test compound in combination with EM (64 μg / mL) on the pmexY strain was evaluated as “−” when the EC ₅₀ was 128 μg / mL or more in the experiment using the pmexY strain, and 8 μg When it was not less than 128 μg / mL, it was evaluated as “+”, and when it was less than 8 μg / mL, it was evaluated as “++”.

  Table 1 shows the results of evaluating the growth inhibitory effect of each test compound. In Table 1, the “vector strain growth inhibition” column shows the evaluation results of the growth inhibition effect of the test compound alone against the vector strain, and the “pmexB strain growth inhibition” column shows the EM (8 μg / mL) combination with the pmexB strain. The evaluation result of the growth inhibitory effect is shown in the column “pmexY strain growth inhibition”, and the evaluation result of the growth inhibitory effect in combination with EM (64 μg / mL) for the pmexY strain is shown.

  As a result, the compounds represented by the general formula (I), such as H-35, H-39, H-46, H-47, H-50, and H-51, are also H-31, H- Similar to the compounds of 32, H-34, H-35, and H-37, both pmexX strain and pmexY strain had a high growth inhibitory effect under combined administration with EM. From these results, it is clear that the compounds according to the present invention remarkably enhance the susceptibility of both strains to EM by inhibiting the excretion of MexB and MexY, which are multidrug efflux pumps of pmexB and pmexY strains. It is. That is, it was shown that the compound according to the present invention is useful as an inhibitor for the RND type multidrug efflux pump.

  On the other hand, H-8, H-27 to H-30, H-33, H-38, H-41, H-42, H-45, H-, which are not compounds represented by the general formula (I). The 48 and H-49 compounds were unable to inhibit the growth of either or both pmexB and pmexY strains. That is, it was found that these compounds cannot simultaneously inhibit the drug excretion ability of MexB and MexY.

[Test Example 2] Evaluation of combined effect with antibacterial agent against multidrug-resistant Pseudomonas aeruginosa Using the H-31 compound produced in Example 1, the combined effect with antibacterial agent against multidrug-resistant Pseudomonas aeruginosa Examined. As the multidrug resistant Pseudomonas aeruginosa, 55 clinical isolates collected from domestic medical institutions were used. As a combined antibacterial agent, ciprofloxacin (CPFX) which is a quinolone series, imipenem (IPM) which is a carbapenem series, amikacin (AMK) which is an aminoglycoside series, or aztreonam (AZT) which is a monobactam series was used. As a comparative control, ABI-PP (D13-9001), a known MexB inhibitor, was used.

Specifically, the minimum inhibitory concentration (MIC) of the combined antibacterial agent for 55 clinical isolates in the presence of the H-31 compound was determined.
First, the H-31 compound dissolved in DMSO in an MH (Mueller-Hinton) agar medium containing various antibacterial agents at various concentrations by the agar plate medium dilution method has a concentration of 32 μg in the agar medium. A plate medium was prepared so as to be / mL. Similarly, a plate medium in which ABI-PP dissolved in DMSO is added to an MH agar medium containing various antibacterial agents at various concentrations so that the concentration in the agar medium is 32 μg / mL; A plate medium was prepared by adding the same amount of DMSO as a solvent to an MH agar medium containing any antibacterial agent at various concentrations.
Next, each plate medium was cultured overnight at 37 ° C. in MH broth, then 1 μL of each clinical isolate diluted to a concentration of 10 ⁵ CFU / μL in the same liquid medium was inoculated and cultured at 37 ° C. for 20 hours. Later, the minimum combined antibacterial agent concentration at which no bacterial growth was inhibited and colonies were observed was defined as MIC.

  FIGS. 7 to 10 show graphs of the cumulative distribution of MIC for clinically separated multidrug-resistant Pseudomonas aeruginosa strains 55 of the combined antibacterial agent in each plate medium. FIG. 7 shows the results when ciprofloxacin was used together, FIG. 8 shows the results when imipenem was used together, FIG. 9 shows the results when amikacin was used together, and FIG. 10 shows the results when used together with aztreonam. Is the result of As a result, when the concomitant antibacterial agent is amikacin (AMK) which is an aminoglycoside antibacterial agent (FIG. 9), although the effectiveness is not recognized, ciprofloxacin whose concomitant antibacterial agent is quinolone In the case of (CPFX: FIG. 7), imipenem (IPM: FIG. 8) which is a carbapenem series, or aztreonam (AZT: FIG. 10) which is a monobactam series, de-drug of multidrug-resistant Pseudomonas aeruginosa by H-31 compound Tolerance was observed. In particular, the effect of the H-31 compound in the case of AZT was extremely remarkable as that of ABI-PP. The effect of the H-31 compound in the case of CPFX was much greater than that of ABI-PP. The effect on IPM was small compared to the former two, but this was thought to be due to the large contribution of resistance factors other than the drug efflux pump to the bicyclic β-lactam antibacterial agent. For example, since the contribution of β-lactamase is strongly suggested, it can be easily estimated that a combination of a bicyclic β-lactam antibacterial agent and a β-lactamase inhibitor is preferable. As described above, the compound according to the present invention functions as a dual inhibitor that simultaneously inhibits the excretion of MexB and MexY, which are the main multidrug efflux pumps, against clinical isolates of multidrug resistant Pseudomonas aeruginosa. Thus, it has been proved that a combination effect with an antibacterial agent is brought about by making it resistant to de-drug.

Claims (12)

The following general formula (I)

[In Formula (I), R ¹ represents a thiazole group, an adamantyl group, or an alkyl group having 3 to 6 carbon atoms in which one hydrogen atom may be substituted with an alkyl group having 1 to 4 carbon atoms. X represents a hydrogen atom or a halogen atom, R ² represents an amino group or a hydrogen atom, n represents an integer of 1 to 4, and R ³ represents one hydrogen atom substituted with an amidino group. An amino group or an imidazole group which may be present. ]
Or a physiologically acceptable salt thereof, or a solvate thereof. R ¹ is a thiazole group in which one hydrogen atom may be substituted with an alkyl group having 1 to 4 carbon atoms, X is a hydrogen atom, a chlorine atom, or a bromine atom, and R ² is an amino group The compound according to claim 1, or a physiologically acceptable salt thereof, or a solvate thereof. The compound according to claim 1 or 2, wherein R ³ is an unsubstituted amino group or an amino group in which one hydrogen atom is substituted with an amidino group, and n is 3 or 4, or a physiologically acceptable compound thereof Salts, or solvates thereof. The compound represented by the general formula (I) is represented by the following formulas (H-31), (H-32), (H-34) to (H-37), (H-39), (H-46). , (H-47), (H-50), or (H-51)

The compound according to any one of claims 1 to 3, or a physiologically acceptable salt thereof, or a solvate thereof.   One or two or more of the compounds according to any one of claims 1 to 4 or a physiologically acceptable salt thereof or a solvate thereof as an active ingredient, and drug discharge of a multidrug discharge pump A multidrug efflux pump inhibitor having activity to inhibit the ability.   The multidrug efflux pump inhibitor according to claim 5, which has an activity of inhibiting both the drug excretion ability of MexB and the drug excretion ability of MexY.   A pharmaceutical composition comprising one or more of the compounds according to any one of claims 1 to 4 or a physiologically acceptable salt thereof, or a solvate thereof as an active ingredient.   The pharmaceutical composition according to claim 7, which is used for prevention and / or treatment of a bacterial infection.   The pharmaceutical composition according to claim 8, further comprising one or more antibacterial agents.   The pharmaceutical composition according to claim 8 or 9, wherein the pathogenic bacterium of the bacterial infection is a gram-negative bacterium.   The pharmaceutical composition according to claim 8 or 9, wherein the pathogenic bacterium of the bacterial infection is Pseudomonas aeruginosa.   The pharmaceutical composition according to any one of claims 8 to 11, wherein the pathogenic bacterium of the bacterial infection is a bacterium that has acquired resistance to an antibacterial agent.

Images