Metallo-beta-lactamase inhibitors

Abstract

The present invention provides: a compound represented by formula (I), formula (II), or formula (IV), or a pharmaceutically acceptable salt thereof; and a pharmaceutical composition that is to be used for inhibiting metallo-β-lactamase and that contains said compound or a pharmaceutically acceptable salt thereof.

Description

[Technical Field]

[0001] The present invention relates to a compound having a β-lactam structure, a pharmaceutical composition for use in metallo-β-lactamase inhibition, and a method for treating infections caused by β-lactam-resistant bacteria. [Background technology]

[0002] In recent years, many reports have described infectious disease-causing bacteria that have acquired resistance to β-lactam antibiotics, and the difficulty of treating them has become a problem. The most prominent resistance mechanism is the production of β-lactamases, which are involved in the degradation and inactivation of β-lactam antibiotics. β-lactamases are classified into classes A, B, C, and D based on their primary amino acid sequence. β-lactamases belonging to class B are called metallo-β-lactamases and differ from serine β-lactamases of other classes (classes A, C, and D) that have serine residues in their active site, in that they are metallo-β-lactamases containing zinc in their active site.

[0003] Metallo-β-lactamases exhibit broad substrate specificity, and bacteria producing them pose a threat because they develop resistance to many clinically important β-lactam antibiotics. For example, they hydrolyze carbapenem antibiotics, which are relatively stable against serine β-lactamase. Furthermore, metallo-β-lactamases have been identified in multiple bacterial species, and the production of metallo-β-lactamases by Pseudomonas aeruginosa (Pseudomonas aeruginosa) leading to multidrug resistance is a particular problem. Currently, β-lactamase inhibitors used are those effective against serine β-lactamase, such as clavulanic acid, sulbactam, and tazobactam, but no inhibitors effective against metallo-β-lactamase have yet been put into practical use.

[0004] The isolation and purification of metallo-β-lactamases have been reported (Non-Patent Document 5). Furthermore, studies have been conducted on metallo-β-lactamase inhibitors, such as succinic acid derivatives, maleic acid derivatives, and phthalic acid derivatives (Patent Documents 1-9). In addition, various compounds with inhibitory activity against metallo-β-lactamases have been reported (Patent Documents 10-22 and Non-Patent Documents 1-4). [Prior art documents] [Patent Documents]

[0005] [Patent Document 1] Special Publication 2003-513890 A [Patent Document 2] Special Publication 2003-527332 A [Patent Document 3] Japanese Patent Publication No. 2016-179964 A [Patent Document 4] Japanese Patent Publication No. 2008-115183 A [Patent Document 5] Japanese Patent Publication No. 2009-040743 A [Patent Document 6] Japanese Patent Publication No. 2013-032361 A [Patent Document 7] Japanese Patent Publication No. 2013-100289 A [Patent Document 8] WO2007 / 034924 [Patent Document 9] WO2008 / 016007 [Patent Document 10] WO2013 / 015388 [Patent Document 11] Special Publication 2016-538244 A [Patent Document 12] Japanese Patent Publication No. 2000-136133 A [Patent Document 13] Japanese Patent Publication No. 2000-143511 A [Patent Document 14] Special Publication 2005-525399 A [Patent Document 15] Special Publication No. 2000-504311 A [Patent Document 16] Special Publication 2018-515481 A [Patent Document 17] Special table 11-514981 A [Patent Document 18] Special Publication 2016-520582 A [Patent Document 19] Japanese Patent Publication No. 2017-101027 A [Patent Document 20] Japanese Patent Publication No. 2017-132766 A [Patent Document 21] Japanese Patent Publication No. 2019-195315 A [Patent Document 22] Japanese Patent Publication No. 2000-336075 A [Non-patent literature]

[0006] [Non-Patent Document 1] Yan, YH. et al., Med Res Rev. 2020; 40:1558-1592.; [Non-Patent Document 2] Palacios, AR., et al., Biomolecules 2020, 10, 854; doi:10.3390 / biom10060854; [Non-Patent Document 3] Jackson, AC. et al., Chem Med Chem 2021, 16, 654-661; [Non-Patent Document 4] Wachino J. et al., mBio 2020, 11:e03144-19. [Non-Patent Document 5] Osano E., et al., Antimicrobial Agents and Chemotherapy, Jan. 1994, 71-78 [Overview of the project] [Problems that the invention aims to solve]

[0007] New means are required for the treatment of infections caused by β-lactam-resistant bacteria, particularly for the treatment of infections caused by resistant bacteria that produce metallo-β-lactamase. The object of the present invention is to provide a metallo-β-lactamase inhibitor that can be used to suppress the inactivation of β-lactam antibiotics by inhibiting metallo-β-lactamase.

Means for Solving the Problems

[0008] The inventors of the present invention have found a compound having metallo-β-lactamase inhibitory activity and have completed the present invention. The present specification includes the disclosure of the following inventions.

[0009] [1] Formula (I) or formula (II):

[0010]

Chemical formula

[0011] [ka] A polyamine group represented by one of the following, where ● indicates the bond position. A compound represented by, or a pharmaceutically acceptable salt thereof.

[0012] [2]R 1 However, X 1 Phenyl, X which may be substituted with one or more substituents selected from 1 A 5 or 6-membered heteroaryl, or X, which may be substituted with one or more substituents selected from 3 C may be substituted with one or more substituents selected from 6-10 A cycloalkanedienyl compound, as described in [1], or a pharmaceutically acceptable salt thereof.

[0013] [3]R 4However, a hydrogen atom, or X 4 C is substituted with one substituent selected from 1-6 A compound described in [1] or [2], or a pharmaceutically acceptable salt thereof, which is alkyl.

[0014] [4] A compound according to any of [1] to [3], or a pharmaceutically acceptable salt thereof, wherein Ra is a polyamine group represented by formula IIIa, formula IIIh, or formula IIIi.

[0015] [5]R 2 The compound is a hydrogen atom, as described in any of [1] to [4], or a pharmaceutically acceptable salt thereof.

[0016] [6] A compound represented by formula II, one of the compounds described in [1] to [5], or a pharmaceutically acceptable salt thereof.

[0017] [7]R 5 However, hydrogen atoms, halogen atoms, C 1-6 Alkyl, C 1-6 Alkoxy, or C 2-6 A compound that is an alkenyl, as described in any of [1] to [6], or a pharmaceutically acceptable salt thereof.

[0018] A pharmaceutical composition comprising any of the compounds described in [8][1] to [7], or a pharmaceutically acceptable salt thereof.

[0019] [9] The pharmaceutical composition described in [8] for use in the treatment of infections caused by β-lactam antibiotic-resistant bacteria.

[0020]

[10] The pharmaceutical composition according to [8] or [9], wherein the resistant bacterium is a metallo-β-lactamase expressing bacterium.

[0021]

[11] A pharmaceutical composition according to any one of [8] to

[10] for use in combination with a β-lactam antibiotic.

[0022] A metallo-β-lactamase inhibitor comprising any of the compounds described in

[12] [1] to [7], or a pharmaceutically acceptable salt thereof.

[0023]

[13] A method for treating an infection caused by a β-lactam antibiotic-resistant bacterium, comprising administering a compound described in any of [1] to [7], or a pharmaceutically acceptable salt thereof, to a subject in need of treatment.

[0024]

[14] The method according to

[13] , further comprising administering a therapeutically effective dose of a β-lactam antibiotic to the subject.

[0025] This specification further includes the disclosure of the following inventions:

[0026] [A-1] Equation (I), Equation (II), or Equation (IV):

[0027] [ka] [In the formula, Q is a direct bond or base: -N(-R 2 )-CH(-R 1 )-C(=O)-, and the nitrogen atom of the group is linked to a carbonyl group shown in formula (I) or formula (II); R 1 X 1 Phenyl, X which may be substituted with one or more substituents selected from 1 A 5 or 6-membered heteroaryl, X which may be substituted with one or more substituents selected from 2 C may be substituted with one or more substituents selected from 1-10 Alkyl, X 2 C may be substituted with one or more substituents selected from 2-10 Alkenil, X 2 C may be substituted with one or more substituents selected from 2-10 Alkinil, X 2 C may be substituted with one or more substituents selected from 3-10 Cycloalkyl, or X 3C optionally substituted with one or more substituents selected from 6-10 is cycloalkanediyl; R 2 is a hydrogen atom or C 1-6 alkyl; R 3 is a hydrogen atom or C 1-6 alkyl; R 4 is a hydrogen atom, X 4 C optionally substituted with one or more substituents selected from 1-6 alkyl, or 5- or 6-membered non-aromatic heterocyclyloxy optionally substituted with one or more substituents selected from X 5 (where the heterocyclyl of the non-aromatic heterocyclyloxy may be fused to a benzene ring); R 5 is a hydrogen atom, a halogen atom, C 1-6 alkyl, C 1-6 alkoxy, C optionally substituted with R 6 2-6 alkenyl, or methyl substituted with R 7 [[ID=​​​​​​​​​​​​​​​​​​​​​​​​​​Pyridinium-1-yl, or X, which may be substituted with 5 It is 1-methylpyrrolidinium-1-yl, which may be substituted with; X 1 is amino, hydroxy, C 1-6 Alkyl, C 1-6 An alkoxy, a halogen atom, or a phenyl which may be substituted with one or more halogen atoms or hydroxyls; X 2 is amino, hydroxy, C 1-6 Alkyl, halogen atom, carboxy, or (C 1-6 It is an alkoxy carbonyl; X 3 C 1-6 Alkyl or halogen atoms; X 4 is, (C 1-6 Alkyl)carbonyloxy, or (C 1-6 It is an alkoxy carbonyl oxy; X 5 R 8 Even if replaced by C 1-6 Alkyl or carbamoyl; X 6 C 1-6 alkyl, halogen atom, (C 1-6 It is an alkoxy)carbonyl or carboxyl; R 8 is hydroxy, sulfo, or carboxy; R 9 is a hydrogen atom or a methyl atom; Ra is given by the following formula:

[0028] [ka] A polyamine group represented by one of the following, where ● indicates the bond position; Rb is given by the following formula: -Q 1 -NR 10 CO-Ra, -Q 1 -N=C-NR 11 CO-Ra,

[0029] [ka] Selected from the bases represented by, Q 1 C 2-6 It is alkylene, R 10 C is a hydrogen atom. 1-6 Alkyl, or -CH=NH, R 11 and R 12 These are, independently, hydrogen atoms, or C 1-6 It is alkyl, R 13 C is a hydrogen atom. 1-6 Alkyl, -CH2NHSO2NH2, or -CONR 14 R 15 And here alkyl is -NR 14 R 15 They may also be substituted with one or more substituents selected from hydroxyl, R 14 C is a hydrogen atom. 1-6 Alkyl, or X 6 A phenyl which may be substituted with one or more substituents selected from, R 15 is a hydrogen atom, or C 1-6 It is alkyl. A compound represented by, or a pharmaceutically acceptable salt thereof.

[0030] [A-2]R 1 However, X 1 Phenyl, X which may be substituted with one or more substituents selected from 1 A 5 or 6-membered heteroaryl, or X, which may be substituted with one or more substituents selected from 3 C may be substituted with one or more substituents selected from 6-10 A cycloalkanedienyl compound, as described in [A-1], or a pharmaceutically acceptable salt thereof.

[0031] [A-3]R 4However, a hydrogen atom, or X 4 C is substituted with one substituent selected from 1-6 A compound described in [A-1] or [A-2] that is alkyl, or a pharmaceutically acceptable salt thereof.

[0032] [A-4] A compound according to any of [A-1] to [A-3], or a pharmaceutically acceptable salt thereof, wherein Ra is a polyamine group represented by formula IIIa, formula IIIh, or formula IIIi.

[0033] [A-5]R 2 A compound listed in any of [A-1] to [A-4], wherein the compound is a hydrogen atom, or a pharmaceutically acceptable salt thereof.

[0034] [A-6] A compound represented by formula II, one of the compounds listed in [A-1] to [A-5], or a pharmaceutically acceptable salt thereof.

[0035] [A-7]R 5 However, hydrogen atoms, halogen atoms, C 1-6 Alkyl, C 1-6 Alkoxy, or C 2-6 A compound that is an alkenyl, as described in any of [A-1] to [A-6], or a pharmaceutically acceptable salt thereof.

[0036] [A-8] Rb is given by the following equation:

[0037] [ka] A compound selected from the groups represented by [A-1] to [A-7], or a pharmaceutically acceptable salt thereof.

[0038] [A-9] Rb is given by the following equation:

[0039] [ka] A compound selected from the groups represented by [A-1] to [A-8], or a pharmaceutically acceptable salt thereof.

[0040] A pharmaceutical composition comprising any of the compounds listed in [A-10], [A-1], to [A-9], or a pharmaceutically acceptable salt thereof.

[0041] [A-11] The pharmaceutical composition described in [A-10] for use in the treatment of infections caused by β-lactam antibiotic-resistant bacteria.

[0042] [A-12] The pharmaceutical composition according to [A-10] or [A-11], wherein the resistant bacterium is a metallo-β-lactamase expressing bacterium.

[0043] [A-13] A pharmaceutical composition according to any of [A-10] to [A-12] for use in combination with a β-lactam antibiotic.

[0044] A metallo-β-lactamase inhibitor comprising a compound listed in any of [A-14], [A-1], to [A-9], or a pharmaceutically acceptable salt thereof. [Effects of the Invention]

[0045] The present invention provides an inhibitor of metallo-β-lactamase, and further provides a novel means for treating infections caused by drug-resistant bacteria that produce metallo-β-lactamase. [Brief explanation of the drawing]

[0046] [Figure 1] Figure 1 shows the results of purification of the reaction solution by reverse-phase HPLC in Example 1. [Figure 2] Figure 2 shows the results of confirming the molecular weight of the target compound by liquid chromatography-mass spectrometry (LC-MS). [Figure 3] Figure 3 shows the results of quantifying the remaining meropenem in Test Example 1 using tandem mass spectrometry. [Figure 4]Figure 4 is a graph showing the results of quantifying the peak area of ​​meropenem detected in Figure 3. [Figure 5] This graph shows the results of quantifying the remaining meropenem in Test Example 2 using tandem mass spectrometry. [Figure 6] The measurement results for Test Example 3 are shown. Complex formation between DTPA-cephalexin and zinc is confirmed from these results. [Figure 7] This graph shows the results of quantifying the remaining meropenem in Test Example 4 using tandem mass spectrometry. [Figure 8] Figure 8 is a graph showing the susceptibility of IMP-1 expressing E. coli to meropenem when 25 μM of DTPA-cephalexin or 25 μM of EDTA is added. [Figure 9] Figure 9 is a graph showing the susceptibility of IMP-1 expressing Escherichia coli to meropenem after the addition of 25 μM NOTA-cephalexin. [Figure 10] Figure 10 shows the MS spectrum of DTPA-cefaclor (compound 2). [Figure 11] Figure 11 shows the MS spectrum of DTPA-cefrazine (compound 3). [Figure 12] Figure 12 shows the MS spectrum of DTPA-amoxicillin (compound 4). [Figure 13] Figure 13 shows the MS spectrum of NOTA-GA-cefrazine (compound 5). [Figure 14] Figure 14 shows the MS spectrum of DTPA-ADCA (compound 6). [Figure 15] Figure 15 shows the MS spectrum of DTPA-cephalexin (compound 7). [Figure 16] Figure 16 shows the procedure for treating mice in Test Example 6. [Figure 17] Figure 17 is a graph showing the survival rate of infected mice after drug administration in Test Example 6. [Figure 18] Figure 18 is a graph showing the test results confirming the inhibitory effect of compound 8 on the growth of multidrug-resistant Pseudomonas aeruginosa strains in test example 8. [Figure 19] Figure 19 is a graph showing the results of a cytotoxicity test of the compound of the present invention in Test Example 9. [Figure 20] Figure 20 is a graph showing the test results in Test Example 10, which confirmed the effect of DTPA-ADCA on enhancing the antibacterial activity of meropenem against IMP-1 expressing Escherichia coli (clinical isolate). [Figure 21] Figure 21 shows the MS spectrum of NODA-GA-bound doripenem (compound 8). [Modes for carrying out the invention]

[0047] In one aspect, the present invention relates to formula (I), formula (II), or formula (IV):

[0048] [ka] The present invention provides compounds represented by formula (I), or pharmaceutically acceptable salts thereof. Here, the compounds represented by formula (I) include the compounds represented by formulas (Ia) and (Ib) below. The compounds represented by formula (II) also include the compounds represented by formulas (IIa) or (IIb) below.

[0049] [ka] In one aspect, the present invention relates to formula (I) or formula (II):

[0050] [ka] The present invention provides compounds represented by formula (Ia), formula (Ib), formula (IIa), or formula (IIb):

[0051] [ka] The present invention provides a compound represented by [formula], or a pharmaceutically acceptable salt thereof.

[0052] In one embodiment of the present invention, Rb is given by the following formula: -Q 1 -NR 10 CO-Ra, -Q 1 -N=C-NR 11 CO-Ra,

[0053] [ka] Selected from the bases represented by

[0054] In one aspect of the present invention, the following compounds are examples of compounds represented by formula IV.

[0055] [ka]

[0056] Here R 13 Examples include hydrogen atoms, (3-carboxyphenyl)aminocarbonyl, dimethylaminocarbonyl, aminosulfonylaminomethyl, and 3-aminomethyl-2-hydroxypropyl. More specifically, the following compounds are examples:

[0057] [ka]

[0058] In this specification, "C 1-10 "Alkyl" refers to linear, branched, cyclic, or partially cyclic alkyl groups having 1 to 10 carbon atoms, such as methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, t-butyl, n-pentyl, 3-methylbutyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, n-hexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3-ethylbutyl, 2-ethylbutyl, n-heptyl, n-octyl, and n-nonyl. This includes n-decanyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl, and for example, C 1-4 Alkyl and C 1-3 Alkyl compounds are also included.

[0059] In this specification, "C 1-6 "Alkyl" refers to linear, branched, cyclic, or partially cyclic alkyl groups having 1 to 6 carbon atoms, and includes, for example, methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, t-butyl, n-pentyl, 3-methylbutyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, n-hexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3-ethylbutyl, and 2-ethylbutyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cyclopropylmethyl, for example, C 1-4 Alkyl and C 1-3 Alkyl compounds are also included.

[0060] In this specification, "C 1-6 "Alkoxy" refers to an alkyloxy group having an alkyl group with 1 to 6 carbon atoms as defined in the alkyl portion [-O-(C 1-6 This means alkyl), and includes, for example, methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, s-butoxy, i-butoxy, t-butoxy, n-pentoxy, 3-methylbutoxy, 2-methylbutoxy, 1-methylbutoxy, 1-ethylpropoxy, n-hexyloxy, 4-methylpentoxy, 3-methylpentoxy, 2-methylpentoxy, 1-methylpentoxy, 3-ethylbutoxy, cyclopentyloxy, cyclohexyloxy, cyclopropylmethyloxy, etc., for example, C 1-4 Alkoxy and C 1-3 Alkoxy compounds are also included. Furthermore, in this specification, "C 1-4 For example, C 1-3 This also includes alkoxy compounds.

[0061] In this specification, "C 2-10 "Alkenyl" refers to a linear, branched, cyclic, or partially cyclic alkenyl group having 2 to 10 carbon atoms, and having one or more, preferably 1 to 3, and more preferably 1 double bond. 2-10 Examples of alkenyls include vinyl, 2-propenyl, 1-propenyl, 1-methylvinyl, 3-butenyl, 2-butenyl, and 1-butenyl.

[0062] In this specification, "C 2-6 "Alkenyl" refers to a linear, branched, cyclic, or partially cyclic alkenyl group having 2 to 6 carbon atoms, and having one or more, preferably 1 to 3, and more preferably 1 double bond. 2-10 Examples of alkenyls include vinyl, 2-propenyl, 1-propenyl, 1-methylvinyl, 3-butenyl, 2-butenyl, and 1-butenyl.

[0063] In this specification, "C 2-10 "Alkynyl" refers to a linear, branched, cyclic, or partially cyclic alkynyl group having 2 to 10 carbon atoms, and the alkynyl group has one or more, preferably 1 to 3, and more preferably 1 triple bond. 2-6 Examples of alkynyls include ethynyl, 2-propynyl, 1-propynyl, 3-butynyl, 2-butynyl, and 1-butynyl.

[0064] In this specification, "C 6-10 A "cycloalkanedienyl" refers to a cyclic alkenyl group with 6 to 10 carbon atoms and two double bonds. Examples include 1-cyclohexa-1,4-dienyl and 2-cyclohexa-1,4-dienyl.

[0065] In this specification, "C 3-10 "Cycloalkyl" refers to a cyclic alkyl group having 3 to 10 carbon atoms. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0066] In this specification, "C 3-7 "Cycloalkyl" refers to a cyclic alkyl group having 3 to 7 carbon atoms. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

[0067] In this specification, "(C 1-6 "Alkyl)carbonyl" refers to the C that has already been defined as the alkyl portion. 1-6 This refers to an alkylcarbonyl group having an alkyl group, such as methylcarbonyl, ethylcarbonyl, tert-butylcarbonyl, and (C 1-3 This includes alkyl and carbonyl groups.

[0068] In this specification, "(C 1-6 "Alkoxy)carbonyl" refers to the alkoxy portion which has already been defined as C 1-6 This refers to an alkoxycarbonyl group having an alkoxy group, such as methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, and (C 1-3 It contains alkoxycarbonyl compounds, etc.

[0069] In this specification, "(C 1-6 "(alkoxy)carbonyloxy" means (C 1-6 The (alkoxy)carbonyl portion has already been defined as (C 1-6 This refers to an alkoxycarbonyl group having an alkoxycarbonyl group, such as methoxycarbonyloxy, ethoxycarbonyloxy, tert-butoxycarbonyloxy, and (C 1-3 It contains alkoxy (carbonyl)oxy, etc.

[0070] In this specification, "5- or 6-membered heteroaryl ring" is not particularly limited to any aromatic heterocyclic group of a 5- or 6-membered ring containing one or more heteroatoms selected from oxygen, nitrogen, and sulfur atoms, for example, 1 to 4 or 1 to 3 heteroatoms. The heteroaryl group may have carbonyl groups as its ring constituent carbons. Examples include pyridyl, pyrimidyl, pyridazinyl, pyrazyl, furanyl(furyl), thiophenyl(thienyl), oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, and 5-oxo-2,5-dihydro-1,2,4-triazine.

[0071] In this specification, "5- or 6-membered ring heteroaryloxy" refers to a 5- or 6-membered ring heteroaryloxy having a 5- or 6-membered ring heteroaryl as defined above. Examples include pyridyloxy, pyrimidyloxy, pyridadinioxyl, pyrazyloxy, furyloxy (furyloxy), thiophenyloxy (thienyloxy), oxazolyloxy, isoxazolyloxy, oxadiazolyloxy, thiazolyloxy, isothiazolyloxy, thiadiazolyloxy, pyrrolyloxy, imidazolyloxy, pyrazolyloxy, triazolyloxy, tetrazolyloxy, and 5-oxo-2,5-dihydro-1,2,4-triazineoxy.

[0072] In this specification, "5- or 6-membered ring heteroarylsulfanil" means a 5- or 6-membered ring heteroarylsulfanil [(5- or 6-membered ring heteroaryl)-S-] having a 5- or 6-membered ring heteroaryl as defined above. Examples include pyridylsulfanil, pyrimidylsulfanil, pyridadinylsulfanil, pyrazylsulfanil, furylsulfanil (furylsulfanil), thiophenylsulfanil (thienylsulfanil), oxazolylsulfanil, isoxazolylsulfanil, oxadiazolylsulfanil, thiazolylsulfanil, isothiazolylsulfanil, thiadiazolylsulfanil, pyrrolylsulfanil, imidazolylsulfanil, pyrazolylsulfanil, triazolylsulfanil, tetrazolylsulfanil, and 5-oxo-2,5-dihydro-1,2,4-triazinesulfanil.

[0073] In this specification, "5 or 6-membered non-aromatic heterocyclyloxy" means a non-aromatic heterocyclyloxy comprising a non-aromatic heterocyclic group containing one or more heteroatoms selected from nitrogen, oxygen, and sulfur atoms, for example, 1 to 4 or 1 to 3 heteroatoms, as a 5 or 6-membered non-aromatic heterocyclyl. Examples include tetrahydrofuranyloxy, dihydrofuranyloxy, pyrrolidinyloxy, piperidinyloxy, piperazinyloxy, and morpholinyloxy.

[0074] Examples of halogen atoms include fluorine, chlorine, bromine, and iodine.

[0075] In this specification, sulfo represents the group -SO2OH.

[0076] In this specification, X 1 ~X 5 If the molecule may be substituted with one or more substituents selected from the following, the number of substituents is 1 to 4, or 1 to 3, or 1 or 2, or 1. Furthermore, if multiple substituents exist, they may be identical or different.

[0077] In this specification, "pharmaceutically acceptable salt" is not particularly limited to any salt that can be used as a pharmaceutical. Examples of salts formed by the compounds of the present invention with a base include salts with inorganic bases such as sodium, potassium, magnesium, calcium, and aluminum; and salts with organic bases such as methylamine, ethylamine, and ethanolamine. The salt may also be an acid addition salt, and specific examples of such salts include mineral acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, and phosphoric acid; and acid addition salts with organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, and ethanesulfonic acid.

[0078] In one embodiment of the present invention, the compounds represented by formula (I), formula (II), or formula (IV) may exist as pharmaceutically acceptable salts, and some or all of the salt-forming groups, such as carboxyl groups, contained in the compound may form salts. Furthermore, if a cation such as a pyridinium group is present in the compound, the carboxyl group in the molecule may be a counteranion, or another counteranion may be present.

[0079] The atoms contained in the compounds represented by formula (I), formula (II), or formula (IV) (e.g., hydrogen, carbon, oxygen, nitrogen, and sulfur atoms) may be isotopic atoms other than the most abundant isotope in nature, and such isotopic atoms may be radioactive isotopes. That is, according to one aspect of the present invention, compounds of formula (I), formula (II), or formula (IV) as defined herein, or salts thereof, labeled with isotopic atoms are provided. Here, labeling with isotopic atoms is, for example, labeling with radioactive isotopes ( 3 H, 14 C, 32 It may also be P, and from the perspective of ease of compound preparation, 3 Labeling with H is preferred.

[0080] In one embodiment of the present invention, a compound of formula (I), formula (II), or formula (IV), its enantiomer, its diastereomer, or a pharmaceutically acceptable salt thereof is administered as a prodrug and converted in vivo to an active compound.

[0081] In one aspect of the present invention, the pharmaceutical composition may be in various dosage forms, for example, for oral administration, tablets, capsules, powders, granules, pills, liquids, emulsions, suspensions, solutions, alcoholic preparations, syrups, extracts, and elixirs. The pharmaceutical composition of the present invention may be, but is not limited to, parenteral preparations such as injectable preparations such as subcutaneous, intravenous, intramuscular, and intraperitoneal injections; transdermal preparations such as patches, ointments, or lotions; sublingual preparations and oral patches for oral administration; and aerosols for nasal administration. These preparations may be manufactured by known methods commonly used in the pharmaceutical manufacturing process.

[0082] The pharmaceutical composition may contain various commonly used components, such as one or more pharmaceutically acceptable excipients, disintegrants, diluents, lubricants, flavoring agents, colorants, sweeteners, flavoring agents, suspending agents, wetting agents, emulsifiers, dispersants, auxiliary agents, preservatives, buffers, binders, stabilizers, coating agents, etc. The pharmaceutical composition of the present invention may also be in a sustained-release or sustained-release dosage form.

[0083] In one aspect of the present invention, the dosage of the pharmaceutical composition can be appropriately selected depending on the route of administration, the patient's body type, age, physical condition, severity of the disease, time elapsed since the onset of the disease, etc. The pharmaceutical composition of the present invention may contain a therapeutically effective amount and / or a preventively effective amount of the compound of formula (I), formula (II), or formula (IV) described above. In the present invention, the compound of formula (I), formula (II), or formula (IV) described above may generally be used in doses of 1 to 1000 mg / day / adult or 0.01 to 20 mg / day / kg body weight. The pharmaceutical composition may be administered as a single dose or in multiple doses.

[0084] In an orally administered composition containing the compound of the present invention, the content of the compound is, for example, 0.001 to 1000 mg per unit dosage form, specifically 0.01 to 500 mg, and more specifically 0.005 to 100 mg.

[0085] The pharmaceutical composition of the present invention may optionally contain conventionally known components such as colorants, preservatives, fragrances, flavorings, coatings, antioxidants, vitamins, amino acids, peptides, proteins, and minerals (such as iron, zinc, magnesium, and iodine). In one embodiment of the present invention, the pharmaceutical composition may be prepared in a form suitable for oral administration, such as various solid preparations including granules (including dry syrup), capsules (soft capsules, hard capsules), tablets (including chewable tablets), powders, pills, or liquid preparations such as oral solutions (including liquids, suspensions, and syrups).

[0086] Examples of additives for formulation include excipients, lubricants, binders, disintegrants, fluidizers, dispersants, wetting agents, preservatives, viscosity modifiers, pH adjusters, colorants, flavoring and odor-modifying agents, surfactants, and solubilizers. When forming a liquid formulation, thickeners such as pectin, xanthan gum, and guar gum can be added. Furthermore, coating agents can be used to form coated tablets or paste-like gelling agents. Even when preparing other forms, conventional methods should be followed.

[0087] According to one aspect of the present invention, a compound represented by formula (I), formula (II), or formula (IV), or a pharmaceutically acceptable salt thereof, is used as a metallo-β-lactamase inhibitor, which is administered in combination with a β-lactam antibiotic. In one embodiment of the present invention, the metallo-β-lactamase inhibitor is administered simultaneously with, separately from, or sequentially with the β-lactam antibiotic.

[0088] Examples of β-lactam antibiotics include carbapenems, penicillins, cephalosporins, or their prodrugs.

[0089] Examples of carbapenems include imipenem, meropenem, biapenem, doripenem, ertapenem, tebipenem pivoxil, and tomopenem (CS-023). More specific examples of carbapenems are imipenem, meropenem, biapenem, and doripenem.

[0090] Examples of penicillins include benzylpenicillin, phenoxymethylpenicillin, carbenicillin, azidocillin, propicillin, ampicillin, amoxicillin, epicillin, ticalcillin, cyclacillin, pirbenicillin, azurocillin, mezurocillin, sulbenicillin, piperacillin, and other known penicillins, as well as their prodrugs.

[0091] Examples of cephalosporins include cefatolidin, cefaloridine, cephalothin, cefazolin, cephalexin, cefacetril, cefapillin, cephamandol naphate, cefradin, 4-hydroxycephalexin, cefoperazone, latamoxef, cefminox, flomoxef, cefsulodine, ceftazidime, cefuroxime, cefditoren, cefmetazole, cefotaxime, ceftriaxone, cefepime, cefpirome, cefozopran, and their prodrugs.

[0092] According to one aspect of the present invention, another type of antibiotic may be used in addition to the β-lactam antibiotic.

[0093] According to one aspect of the present invention, in addition to a metallo-β-lactamase inhibitor, other β-lactamase inhibitors may be used in combination. Preferred examples include serine β-lactamase inhibitors such as clavulanic acid, sulbactam, or tazobactam.

[0094] In one aspect of the present invention, metallo-beta-lactamase inhibitors are used in the treatment of infections caused by metallo-beta-lactamase-producing strains. Examples of metallo-beta-lactamase-producing strains include Bacillus cereus, Bacteroides fragilis, Escherichia coli, Aeromonas hydrophila, Klebsiella pneumoniae, Pseudomonas aeruginosa, Serratia marcescens, Stenotrophomonas maltophilia, Shigella flexneri, Alcaligenes xylosoxidans, Legionella gormanii, Chryseobacterium meningosepticum, Chryseobacterium indologenes, Acinetobacter baumannii, Citrobacter freundii, and Enterobacter cloacae.

[0095] The dosage of a compound of formula (I), formula (II), or formula (IV), or a pharmaceutically acceptable salt thereof, with an antibiotic can vary over a wide range, but for example, a weight ratio of approximately 1:0.5 to 20, preferably 1:1 to 8, is common.

[0096] Metallo-β-lactamase inhibitors and β-lactam antibiotics can be administered separately, or they can be administered in the form of a single composition containing both active ingredients. In either embodiment, the compound of formula (I), formula (II), or formula (IV) or a pharmaceutically acceptable salt thereof is preferably in the form of a pharmaceutical composition by being combined with an antibiotic and a pharmaceutically acceptable carrier (i.e., a formulation additive).

[0097] Compounds of formula (I), formula (II), or formula (IV) can be synthesized by reacting a β-lactam compound having an amino group with a carboxylic acid corresponding to the polyamine group of formulas IIIa to IIIi. In one embodiment, the reaction can be carried out by reacting a carboxylic acid anhydride with a β-lactam compound, and if the acid anhydride is a divalent acid anhydride, the target compound can be obtained by separating the dimer produced as a by-product. In another embodiment, the target product can be obtained by reacting the carboxylic acid with a β-lactam compound in the presence of a coupling agent.

[0098] The present invention will be illustrated below with examples, but this is not intended to limit the present invention.

[0099] [Example 1] Synthesis of DTPA-bound cephalexin

[0100] [ka]

[0101] Cephalexin (Wako Pure Chemical Industries) was dissolved in a 400 mM aqueous solution of disodium hydrogen phosphate to a concentration of 20 mM. To this solution, anhydrous DTPA powder (Dojin Chemical Laboratories) (see formula below) was added to a final concentration of 20 mM, and the mixture was reacted at 37°C for 30 minutes. The reaction solution was purified by reverse-phase HPLC under the following conditions to obtain compound 1 (Figure 1, yield: 47%).

[0102] Reverse-phase column: YMC-Triat C18 Plus column (4.6 x 250 mm); Column temperature: 35°C; Mobile phase (2 solutions): Mobile phase A (0.1% formic acid aqueous solution), Mobile phase B (acetonitrile); Gradient: Increase from 0.2%B to 40%B over 22 minutes, then maintain at 40%B for 1 minute. Then return to 0.2%B over 1 minute. Flow rate: 0.8 ml / min; Detection: 254 nm; Sample injection volume: 1 ml.

[0103] [ka]

[0104] The elution fraction containing the target compound (Compound 1, molecular weight: 722.72) was recovered (Figure 1) and freeze-dried. The molecular weight of the compounds in the fraction was confirmed by liquid chromatography-mass spectrometry (LC-MS) (Figure 2).

[0105] [Example 2] Synthesis of DTPA-bound cefaclor

[0106] [ka]

[0107] Compound 2 was prepared using cefaclor (Sigma-Aldrich) in the same manner as in Example 1 (yield: 43%). The molecular weight of the target compound was determined by liquid chromatography-mass spectrometry (LC-MS), and the results are shown in Figure 10.

[0108] [Example 3] Synthesis of DTPA-bound cefradin

[0109] [ka]

[0110] Compound 3 was prepared using cefradin (Sigma-Aldrich) in the same manner as in Example 1 (yield: 27%). The molecular weight of the target compound was confirmed by liquid chromatography-mass spectrometry (LC-MS), and the results are shown in Figure 11.

[0111] [Example 4] Synthesis of DTPA-bound amoxicillin

[0112] [ka]

[0113] Compound 4 was prepared using amoxicillin (Fujifilm) in the same manner as in Example 1 (yield: 41%). The molecular weight of the target compound was confirmed by liquid chromatography-mass spectrometry (LC-MS), and the results are shown in Figure 12.

[0114] [Example 5] Synthesis of NOTA-GA-bound cefradin

[0115] [ka]

[0116] Cefrazine (Sigma-Aldrich) was dissolved in a 400 mM aqueous solution of disodium hydrogen phosphate to a concentration of 20 mM. To this solution, powdered NOTA-GA-NHS (2,2'-(7-(1-carboxy-4-((2,5-dioxopyrrolidine-1-yl)oxy)-4-oxobutyl)-1,4,7-triazonan-1,4-diyl)diacetic acid; CheMatech, France) was added to a final concentration of 30 mM, and the mixture was reacted at 37°C for 30 minutes. The reaction solution was purified by reverse-phase HPLC under the following conditions to obtain compound 5 (yield: 85%).

[0117] Reverse-phase column: YMC-Triat C18 Plus column (4.6 x 250 mm); Column temperature: 35°C; Mobile phase (2 solutions): Mobile phase A (0.1% formic acid aqueous solution), Mobile phase B (acetonitrile); Gradient: Increase from 0.2%B to 40%B over 22 minutes, then maintain at 40%B for 1 minute. Then return to 0.2%B over 1 minute. Flow rate: 0.8 ml / min; Detection: 254 nm; Sample injection volume: 1 ml.

[0118] Figure 13 shows the results of confirming the molecular weight of the target compound by liquid chromatography-mass spectrometry (LC-MS).

[0119] [Example 6] Synthesis of DTPA-bound ADCA

[0120] [ka]

[0121] Compound 6 was prepared using 7-aminodesacetoxycephalosporanic acid (7-ADCA, Tokyo Chemical Industry Co., Ltd.) in the same manner as in Example 1 (yield: 58%). The molecular weight of the target compound was confirmed by liquid chromatography-mass spectrometry (LC-MS), and the results are shown in Figure 14.

[0122] [Example 7] Synthesis of NOTA-conjugated cephalexin

[0123] [ka]

[0124] Cephalexin (Wako Pure Chemical Industries) was dissolved in a 400 mM aqueous solution of disodium hydrogen phosphate to a concentration of 30 mM. To this solution, powdered NOTA-NHS (2,2'-(7-(2-((2,5-dioxopyrrolidine-1-yl)oxy)-2-oxoethyl)-1,4,7-triazonan-1,4-diyl)diacetic acid (CheMatech)) was added to a final concentration of 40 mM, and the mixture was reacted at 37°C for 30 minutes. The reaction solution was purified by reverse-phase HPLC under the following conditions to obtain compound 7 (yield: 82%).

[0125] Reverse-phase column: YMC-Triat C18 Plus column (4.6 x 250 mm); Column temperature: 35°C; Mobile phase (2 solutions): Mobile phase A (0.1% formic acid aqueous solution), Mobile phase B (acetonitrile); Gradient: Increase from 0.2%B to 40%B over 22 minutes, then maintain at 40%B for 1 minute. Then return to 0.2%B over 1 minute. Flow rate: 0.8 ml / min; Detection: 254 nm; Sample injection volume: 1 ml.

[0126] Figure 15 shows the results of confirming the molecular weight of the target compound by liquid chromatography-mass spectrometry (LC-MS).

[0127] [Example 8] Synthesis of NODA-GA-conjugated doripenem

[0128] [ka]

[0129] Doripenem (Tokyo Chemical Industries) was dissolved in a 400 mM aqueous solution of disodium hydrogen phosphate to a concentration of 30 mM. To this solution, powdered NODA-GA-NHS(2,2'-(7-(1-carboxy-4-((2,5-dioxopyrrolidine-1-yl)oxy)-4-oxobutyl)-1,4,7-triazonan-1,4-diyl)diacetic acid (CheMatech)) was added to a final concentration of 40 mM, and the mixture was reacted at 37°C for 30 minutes. The reaction solution was purified by reverse-phase HPLC under the following conditions to obtain compound 7 (yield: 75%).

[0130] Reverse-phase column: YMC-Triat C18 Plus column (4.6 x 250 mm); Column temperature: 35°C; Mobile phase (2 solutions): Mobile phase A (0.1% formic acid aqueous solution), Mobile phase B (acetonitrile); Gradient: Increase from 0.2%B to 40%B over 22 minutes, then maintain at 40%B for 1 minute. Then return to 0.2%B over 1 minute. Flow rate: 0.8 ml / min; Detection: 254 nm; Sample injection volume: 1 ml.

[0131] [Test Example 1] Based on the description in Non-Patent Document 5, recombinant metallo-β-lactamase (recombinant IMP-1) was prepared. The IMP-1 gene, "NG_049172" from Database: RefSeq, was incorporated into the vector pET29a (Novagen) and expressed in E. coli BL21 (DE3) (Invitrogen).

[0132] Recombinant IMP-1 was dissolved at a concentration of 0.25 μg / ml in 50 mM sodium phosphate buffer (pH 7.6). Zinc chloride was then added to a concentration of 0.1 μM. This mixture was pre-incubated at 37°C for 10 minutes. DTPA-cephalexin (compound 1) was then added at concentrations of 1, 5, and 10 μM, followed by the addition of 100 μM meropenem (Wako Pure Chemical Industries). The mixture was incubated at 37°C for 1 hour, and the remaining meropenem was quantified by tandem mass spectrometry. Meropenem was further quantified by multiple reaction monitoring. The measurement conditions were as follows.

[0133] Reverse-phase column: YMC-Triat C18 Plus column (2.1 x 50 mm); Column temperature: 45°C; Mobile phase (2 solutions): Mobile phase A (0.1% formic acid aqueous solution), Mobile phase B (acetonitrile); Gradient: Increase from 1%B to 80%B over 10 minutes, then maintain at 80%B for 0.5 minutes. Then return to 1%B over 1 minute. Flow rate: 0.2 ml / min; Sample injection volume: 10 μl. Detection: Multiple reaction monitoring. Parent ion 384.2, child ion 68.1; Measured in positive ion mode.

[0134] The results are shown in Figures 3 and 4. A peak for meropenem is detected around 7.5 minutes in Figure 3. Meropenem is completely degraded when incubated with IMP-1. Adding DTPA-cephalexin then inhibits the degradation of meropenem.

[0135] Figure 4 shows a graph quantifying the peak area of ​​meropenem detected in Figure 3. The peak containing only meropenem is shown as 100%.

[0136] [Test Example 2] Under the same conditions as in Test Example 1, tests were conducted using DTPA, DTPA-cefrazine, and DTPA-cefachlor, and the inhibitory effect of IMP-1 on the degradation of meropenem when each sample was added at a concentration of 1 μM was evaluated. The results are shown in Figure 5.

[0137] [Test Example 3] DTPA-cephalexin (100 μM) and zinc chloride (ZnCl2, 100 μM) were reacted in 50 mM sodium phosphate buffer (pH 7.6) at 37°C for 1 hour. For comparison, DTPA-cephalexin alone was incubated similarly at 37°C for 1 hour. Subsequently, complex formation between DTPA-cephalexin and zinc was confirmed by mass spectrometry. Measurements were performed in negative mode. The results are shown in Figure 6. DTPA-cephalexin was detected with an m / z of 721 (peak 1). On the other hand, when zinc coordinated with DTPA-cephalexin, an m / z of 783 (peak 2) was detected, with the molecular weight of zinc added.

[0138] [Test Example 4] Under the same conditions as in Test Example 1, tests were conducted using DTPA and NOTA-GA-cefrazine, and the inhibitory effect of IMP-1 on the degradation of meropenem when each sample was added at a concentration of 1 μM was evaluated. The results are shown in Figure 7.

[0139] [Test Example 5] Based on the description in Non-Patent Document 5, IMP-1 expressing Escherichia coli was prepared.

[0140] The effects of DTPA-cephalexin and NOTA-cephalexin on the susceptibility of IMP-1 expressing Escherichia coli to carbapenem antibiotics (meropenem) were evaluated using the following method. IMP-1 expressing Escherichia coli was cultured overnight with shaking in LB medium containing 100 μg / ml ampicillin. The resulting bacterial suspension was diluted to 1 / 200th in LB medium to prepare the test suspension, which was then seeded into a 96-well plate. Meropenem (Wako Pure Chemical Industries) was added to this suspension in two step dilutions, starting from a maximum of 2 μM. The bacteria were cultured for 1 day in an incubator at 37°C, and the growth during this time was measured by turbidity (655 nm). The results are shown in Figures 8 and 9.

[0141] Figure 8 shows the susceptibility to meropenem when 25 μM of DTPA-cephalexin or 25 μM of EDTA or DTPA is added. Figure 9 shows the susceptibility to meropenem when 25 μM of NOTA-cephalexin is added. The results labeled "control" in the figures show the results when only meropenem was added. It was confirmed that when DTPA-cephalexin and NOTA-cephalexin were present together, metallo-β-lactamase-expressing bacteria were killed by the action of carbapenem antibacterial agents.

[0142] [Test Example 6] In vivo experiments using bacterial-infected mice were conducted to investigate the drug susceptibility-enhancing effect of DTPA-conjugated cephalexin (compound 1) on pathogenic bacteria. To create an immunocompromised Leukopenia mouse model, 0.1 mL of cyclophosphamide monohydrate (Sigma-Aldrich) prepared in PBS was administered intraperitoneally to 4-week-old male ddY mice (Nippon SLC) at a dose of 250 mg / kg four days before infection. IMP-1 expressing Klebsiella pneumoniae was obtained from clinical isolates provided by Kumamoto University Hospital. The bacteria were cultured overnight in LB medium at 37°C with shaking, then diluted 50-fold in LB medium and cultured again with shaking until the turbidity (600 nm) was 1.5 or higher. The bacteria were centrifuged to remove the medium, washed twice with PBS, and then refractory to 5 × 10⁴ 5 The solution was diluted with PBS to a concentration of CFU / mL. In mouse infection experiments, 0.1 mL (5 × 10⁶) of IMP-1 expressing Klebsiella pneumoniae was administered to each Leukopenia mouse. 4 CFU was administered intraperitoneally to the mice for infection. The treatment group received a mixture of meropenem (Wako Pure Chemical Industries) and compound 1 (DTPA-CEF) at concentrations of 10 mg / kg and 50 mg / kg, respectively, at a dose of 0.1 mL subcutaneously 30 minutes after infection. The control group received either meropenem (10 mg / kg) or the solvent PBS at a dose of 0.1 mL subcutaneously. The viability of the mice was assessed 48 hours after infection. The results are shown in Figure 17. The number of mice in each group was 7 in the PBS group, 4 in the meropenem group, and 4 in the meropenem and DTPA-CEF combined group.

[0143] The Klebsiella pneumoniae expressing IMP-1 showed lethality in the leukopenia model mice, and the survival rate decreased to 14% after 48 hours (Figure 17, PBS administration group). All mice died in the meropenem administration group, but the survival rate improved to 50% in the treatment group administered with meropenem and compound 1 (Figure 17, MEPM administration group, and MEPM + DTPA-CEF administration group).

[0144] [Test Example 7] The effect of NODA-GA-conjugated doripenem (Compound 8) on the carbapenem antibacterial susceptibility of clinically isolated Pseudomonas aeruginosa was evaluated by the following method. Clinically isolated Pseudomonas aeruginosa was cultured overnight with shaking in LB medium. The bacterial solution diluted 1000-fold with LB medium was used as the test bacterial solution and seeded in a 96-well plate. Meropenem (Wako Pure Chemical Industries, Ltd.) was added to this bacterial solution to a concentration of 2 μg / ml. In addition, the effect of NODA-GA-doripenem alone was also examined. The bacteria were cultured for 1 day in an incubator at 37°C, and the growth at that time was measured by turbidity (655 nm). The results are shown in the following table.

[0145]

Table 1

[0146] In Table 1, 〇 indicates that the growth of bacteria was inhibited in all 3 wells. △ indicates that the growth of bacteria was inhibited in 2 out of 3 wells. × indicates that the number of wells in which growth inhibition of bacteria was confirmed was 1 or less. MEPM in Table 1 is the single administration of meropenem (2 μg / ml); NODA-GA-doripenem is the single administration of Compound 8 (20 μM; 15.5 μg / ml), and Combination means the combined use of meropenem (2 μg / ml) and Compound 8 (20 μM; 15.5 μg / ml).

[0147] [Test Example 8] Multidrug-resistant Pseudomonas aeruginosa strain MR4 was cultured with doripenem or compound 8, and the bacterial load after 24 hours was evaluated by turbidity. The results are shown in Figure 18. It was confirmed that the anti-Pseudomonas activity of NODA-GA-conjugated doripenem was significantly improved compared to doripenem alone.

[0148] [Test Example 9] The cytotoxicity of compound 6 (DTPA-conjugated ADCA), compound 1 (DTPA-conjugated cephalexin), and compound 8 (NODA-GA-conjugated doripenem) was evaluated using the 3-(4,5-dimethylthia-2-yl)-2,5-tetrazolium bromide (MTT) method. HeLa cells were used in each well (1 × 10⁶ cells). 4 The cells were dispensed into 96-well plates and cultured overnight at 37°C under 5% carbon dioxide circulation. The test compound was added to these cells in two step dilutions, starting from a maximum of 400 μM. Six hours after compound treatment, the medium was changed, and MTT dissolved in PBS to a concentration of 7.5 mg / mL was added, followed by a further 2-hour reaction. The supernatant was then removed from each well, and isopropanol hydrochloride solution was added to dissolve the formazan crystals. Cytotoxicity was calculated from the difference between the increase in absorption at 490 nm due to formazan elution and the absorbance at 655 nm. The results are shown in Figure 19. The results are expressed with the case without compound addition set to 100%.

[0149] [Test Example 10] The enhancement effect of DTPA-ADCA on the susceptibility of clinical isolates of Escherichia coli expressing IMP-1 to carbapenem antibiotics (meropenem) was evaluated using the following method. Clinical isolates of Escherichia coli were cultured overnight with shaking in LB medium. The resulting bacterial suspension was diluted to 1 / 1000 in LB medium to prepare the test suspension, which was then seeded into a 96-well plate. Meropenem (Wako Pure Chemical Industries) was added to this suspension in two step dilutions, starting from a maximum of 10 μg / ml. The bacteria were cultured for 1 day in an incubator at 37°C, and their growth was measured by turbidity (655 nm). The meropenem concentration at which no bacterial growth was observed was defined as the minimum inhibitory concentration.

[0150] The effect of DTPA-ADCA on the meropenem susceptibility of clinical isolates of Escherichia coli was evaluated using the following method. Clinical isolates of Escherichia coli were cultured overnight with shaking in LB medium. The resulting bacterial suspension was diluted to 1 / 1000th in LB medium to prepare the test suspension, which was then seeded into a 96-well plate. Meropenem (Wako Pure Chemical Industries) was added to this suspension in a range of 0.1 μg / ml to 0.005 μg / ml. DTPA or DTPA-ADCA was then added at a concentration of 20 mM to investigate its effect on bacterial growth. The bacteria were cultured for 1 day in an incubator at 37°C, and their growth was measured by turbidity (655 nm).

[0151] The results are shown in Figure 20. Multidrug-resistant E. coli were cultured with the indicated concentrations of the compounds, and the bacterial load after 24 hours was evaluated by turbidity. Meropenem alone had a minimum inhibitory concentration of 10 μg / ml (left graph). When DTPA-ADCA (20 μM) was used in combination, the minimum inhibitory concentration of meropenem decreased to 10 μg / ml, and the antibacterial activity was enhanced 100-fold. DTPA alone did not show such an effect (right graph).

Claims

1. Formula (IV): 【Chemistry 1】 [In the formula, R ３ is a hydrogen atom, or C １－６ It is alkyl; R ９ is a hydrogen atom, or a methyl atom; Ra is given by the following formula: 【Chemistry 2】 A polyamine group represented by one of the following, where ● indicates the bond position; Rb is given by the following formula: -Q １ -NR １０ A-00 -Q １ -N=C-NR １１ A-00 【Transformation 3】 Selected from the bases represented by, Q １ is an alkylene C ２－６ and R １０ C is a hydrogen atom. １－６ Alkyl, or -CH=NH, R １１ and R １２ These are, independently, hydrogen atoms, or C １－６ It is alkyl, R １３ C is a hydrogen atom. １－６ Alkyl, -CH ２ NHSO ２ NH ２ , or -CONR １４ R １５ And here alkyl is -NR １４ R １５ They may be substituted with one or more substituents selected from hydroxyl, R １４ C is a hydrogen atom. １－６ Alkyl, or X ６ A phenyl which may be substituted with one or more substituents selected from, R １５ is a hydrogen atom, or C １－６ It is alkyl. A compound represented by, or a pharmaceutically acceptable salt thereof.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein Ra is a polyamine group represented by formula IIIa, formula IIIh, or formula IIIi.

3. Rb is given by the following equation: 【Chemistry 4】 A compound according to claim 1, or a pharmaceutically acceptable salt thereof, selected from the groups represented by .

4. Rb is given by the following equation: 【Transformation 5】 A compound according to claim 1, or a pharmaceutically acceptable salt thereof, selected from the groups represented by .

5. A pharmaceutical composition comprising a compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof.

6. The pharmaceutical composition according to claim 5, for use in the treatment of infections caused by β-lactam antibiotic-resistant bacteria.

7. The pharmaceutical composition according to claim 6, wherein the resistant bacterium is a metallo-β-lactamase expressing bacterium.

8. The pharmaceutical composition according to claim 5, for use in combination with a β-lactam antibiotic.

9. A metallo-β-lactamase inhibitor comprising a compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof.

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