Antimicrobial agent

Abstract

A compound according to Formula (I) or Formula (la) and compositions comprising the same for use as antimicrobial agents: wherein X1, X2, X3, and X4 are each independently selected from: N, O, S; Y1 and Y2 are each independently selected from: N, O, S, C(R a ); M1 and M2 are each a metal center; R1, R2, R3, R a 4, and R5 are each independently selected from: hydrogen, alkyl, alkenyl, aryl, halogen, haloalkyl, haloalkenyl, haloaryl, hydroxyl, alkoxy, carboxylic acid, amino, amido, nitro, or combinations thereof; A1, A2, A3, and A4 are each a bidentate ligand; and rings D1 and D2 each independently comprise one or more heteroatoms selected from N, O, S, C(R a ); wherein the compound is for use as an antimicrobial agent.

Description

TECHNICAL FIELD

[0001] The present invention relates to compounds for use in the treatment of microbial infections, in particular bacterial infections. The invention also describes novel compounds, methods of preparing said compounds and methods of treating diseases and conditions caused by antimicrobial infections. BACKGROUND

[0002] While polypyridyl ruthenium(II) complexes have been the subject of much research as imaging probes and leads for anticancer therapy, initial biological studies focused on their potential as antimicrobial agents. In a work that was ahead of its time in many ways, the Dwyer group demonstrated that lipophilic derivatives of [Ru(phen)3] 2+ (phen = 1,10-phenanthroline) were active against a range of bacteria, particularly Gram-positive bacteria, which were unable to develop resistance to these structures. This work was not pursued further due to the relatively high minimum inhibitory concentrations (MICs) of these complexes compared to contemporary commercial antibacterial agents. However, in the modern context of a rapidly emerging global health crisis due to increasing antimicrobial resistance (AMR), the activity of this class of structures is being revisited.

[0003] It was recently discovered that tethering together [Ru(phen)3] 2+ units using flexible methylene linkers of different lengths resulted in complexes with quite high activity (low MICs). The mechanism of activity of these compounds is still under investigation. While some studies have speculated that they accumulate at the ribosome to cause polysome condensation, others have suggested that cellular uptake and antibacterial activity of this class of compounds is due to their transmembrane ability. However, these systems have shown low activity against Gram-negative bacteria such as E. coli, with one notable exception involving a mononuclear complex. In fact, pathogenic Gram-negative bacteria are a particularly problematic AMR issue. For example, in a recent report, the World Health Organization declared that the post-antibiotic era “is very likely to become a reality in the 21st century” and identified the Gram-negative Pseudomonas aeruginosa, Acinetobacter baumannii and members of the Enterobacteriaceae family (which make up the majority of the ESKAPE group of serious nosocomial infections) as three of the top-priority (critical) pathogens in its “priority pathogens list for R&D of new antibiotics”. The urgency of the situation is exacerbated by the lack of new therapeutic leads: it is over 50 years since a new antibiotic against Gram-negative pathogens was approved and since 2010 only one new compound has entered the antibiotic pipeline through phase 1 trials. This situation has prompted calls for increased chemical diversity in the search for chemical leads. The lack of wholly new molecular “starting point leads” has also been identified as the biggest obstacle to antibiotic discovery.

[0004] There have been some studies of ruthenium complexes as imaging agents for bacterial species, particularly for use in confocal microscopy (see, for example, WO 2009 / 050509). However, to date, no substantially effective ruthenium complex has been found which could form the basis of a new class of antibiotic and / or antimicrobial agent.

[0005] The present invention aims to address or at least ameliorate this problem. SUMMARY

[0006] In a first aspect of the present invention, there is provided a compound according to formula (I) or formula (la):

[0007]

[0008] wherein,

[0009] X1, X2, X3and X4are each independently selected from: N, O, S;

[0010] Y1and Y2are each independently selected from: N, O, S, C(R a );

[0011] M1and M2are each a metal centre;

[0012] R1, R2, R3, R4and R a are each independently selected from: hydrogen, alkyl, alkenyl, aryl, halogen, hydroxyl, alkoxy, carboxylic acid, amino, amido, nitro or combinations thereof; and

[0013] A1, A2, A3and A4are each a bidentate ligand;

[0014] wherein each of rings D1and D2may independently comprise one or more heteroatoms; and

[0015] wherein the compound is for use as an antimicrobial agent.

[0016] The inventors have found that bidentate functionalised complexes according to formula (I) or formula (la) not only can act as effective imaging agents, but also provide significant antimicrobial properties. Without being bound by theory, it is hypothesised that these compounds initially disrupt the cell membrane, attacking both the outer and inner membranes (tearing and "popping", respectively). Furthermore, it is believed that the compounds of formula (I) interact with cardiolipin (a double negative charged lipid on the inner membrane leaflet).

[0017] With respect to the compounds of formula (la), again without being bound by theory, it is believed that these compounds target DNA.

[0018] Typically, the compounds described in the first aspect of this invention are used as antibiotics. While these compounds can be used to treat a wide range of microorganisms, the compounds according to the first aspect of this invention have been found to be remarkably effective against bacteria. This is especially true for Gram-negative bacteria, to which the compounds of the first aspect of this invention are particularly effective. However, it is also contemplated that the claimed compounds may be useful against Gram-positive bacteria.

[0019] There are no particular limitations on the bacteria to which the compounds of the present invention can be targeted. However, generally, the compounds of the first aspect of the present invention are used to treat one or more bacterial species selected from Escherichia coli, Acinetobacter baumannii, Burkholderia cepacia, Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus faecalis, and Streptococcus.

[0020] To avoid ambiguity, the term "alkyl" is intended to be used in its usual sense, covering carbon lengths from C1 to C2. 30 The range includes straight-chain, branched, and cyclic saturated hydrocarbons. The alkyl group is typically straight-chain or branched, and usually straight-chain. While the exact length of the alkyl group can vary, it is typically between C1 and C2. 20 More typically C1 to C 12 More typically, C1 to C8, most typically C1 to C6. Typical alkyl groups are selected from, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, or tert-butyl. Similarly, the term "alkoxy" is intended to be used in its usual sense, that is, the term is the same as "alkyl" above, but the alkyl group is covalently bonded by oxygen atoms (as is known to those skilled in the art).

[0021] To avoid ambiguity, the term "alkenyl" is intended to be used in its usual sense, covering carbon lengths from C2 to C3. 30 This refers to straight-chain, branched, and cyclic partially saturated hydrocarbons within the range. The term "partially saturated" means that at least one C=C bond exists in the structure. The alkenyl group is typically straight-chain or branched, but usually straight-chain. Although the exact length of the alkenyl group can vary, it is typically between C2 and C3. 20 More typically, C2 to C 12 More typically, it ranges from C2 to C8, and most typically from C2 to C6. Typical alkenyl groups are selected from, but are not limited to, vinyl, propenyl, isopropenyl, or butenyl.

[0022] For the avoidance of doubt, each of the “alkyl” and “alkenyl” listed above may optionally be substituted. One or more optional substituents may be present, and typical optional substituents are selected from: halogens, hydroxyl groups, carboxylic acids, amines, amides, nitro groups, or combinations thereof. Most typically, the optional substituents are halogens or hydroxyl groups. In some embodiments, one or more hydrogen atoms may be replaced by halogens.

[0023] For the avoidance of doubt, the term "aryl" is intended to be used in its usual sense to encompass groups of one or more aromatic ring types. The aromatic rings may include eight-membered, seven-membered, six-membered, and five-membered rings. More typically, the ring is a six-membered or five-membered ring, and most typically, the ring is a six-membered ring. Furthermore, the ring may contain one or more heteroatoms to form heteroaryl types. Typical heteroatoms include N, O, and S, more typically N and O, and most typically N. In the use of heteroaryl groups, typically only one heteroatom is present. Generally, the aryl group is selected from: phenyl, cyclopentadienyl, pyridyl, and furanyl.

[0024] For the avoidance of doubt, the “aryl” group listed above may optionally be substituted. One or more optional substituents may be present, and typical optional substituents are selected from: alkyl, alkoxy, alkenyl, halogen, hydroxyl, carboxylic acid, amine, amide, nitro, or combinations thereof. More typically, the optional substituent is alkyl, alkoxy, halogen, or hydroxyl, and even more typically, the optional substituent is alkyl or alkoxy (usually alkyl). In some embodiments, one or more arbitrary hydrogen atoms may be substituted with a halogen.

[0025] For the avoidance of doubt, the term "halogen" is intended to be used in its common sense, generally encompassing fluorine, chlorine, bromine, and iodine. More typically, the halogen is fluorine, chlorine, or bromine; even more typically, fluorine or chlorine; most typically, fluorine.

[0026] Furthermore, the term "metal center" is intended to refer to a metal atom (usually a metal ion) around which a suitable ligand can be bonded to form a complex. There are no particular restrictions on the choice of metal, but metal centers are generally capable of forming complexes with a hexagonal geometry, i.e., capable of forming six bonds. Typically, the metal center is a transition metal.

[0027] The types of X1, X2, X3, and X4 (i.e., whether they are N, O, or S) depend largely on the choice of the metal center (i.e., “M1” and “M2”, respectively) used in the compound of formula (I) or formula (Ia). It is understood that different metals have different affinities for different donor atoms. However, typically, X1, X2, X3, and X4 are each independently N or O. More typically, at least one of X1, X2, X3, and X4 is N; more typically, at least two of X1, X2, X3, and X4 are N; and even more typically, at least three of X1, X2, X3, and X4 are N. However, most typically, X1, X2, X3, and X4 are each N.

[0028] Furthermore, although the types of Y1 and Y2 (i.e. whether N, O, S, C(R) are used) a There are no practical restrictions, but typically Y1 and Y2 are independently selected from N, O, and C(R).a ), and more typically each independently selected from N or C(R) a Typically, at least one of Y1 and Y2 is N. However, the most common case is that both Y1 and Y2 are N.

[0029] For compounds according to formula (Ia), rings D1 and D2 may each independently contain one or more heteroatoms, as typically defined above for X3 and X4. Specifically, X3 and / or X4 may be selected from C, N, and O, more typically C and N, and most typically N. It is possible that two or more heteroatoms are present in rings D1 and / or D2. Typically, the compound has a structure according to formula (Ib):

[0030]

[0031] in,

[0032] A1, A2, M1, R1 to R4, Y1, Y2, and X1 to X4 are defined as above.

[0033] There are no particular limitations on the metal centers M1 and M2 used in the compounds of formulas (I) and (Ia), as long as they can form stable complexes with the ligands of formulas (I) and (Ia). Typical examples of metal centers include, but are not limited to, ruthenium, iridium, osmium, iron, platinum, rhodium, or combinations thereof. Typically, M1 and M2 are selected from ruthenium, iridium, and osmium. Typically, at least one of M1 and M2 is ruthenium or iridium; more typically, it is ruthenium. It is possible that both M1 and M2 are ruthenium. In some embodiments, one or both of the metal centers M1 and M2 may be iridium.

[0034] Typically, R1, R2, R3, and R4 are each independently selected from: hydrogen, alkyl, alkenyl, aryl, halogen, hydroxyl, alkoxy, carboxylic acid, amino, amide, nitro, or combinations thereof. R1, R2, R3, and R4 each represent one to three groups attached to the relevant aromatic ring, as is well known to those skilled in the art, i.e., as shown in formula (II) or formula (IIa):

[0035]

[0036]

[0037] Typically, R1, R2, R3, and R4 are each independently selected from: hydrogen, alkyl, alkenyl, aryl, halogen, hydroxyl, alkoxy, or combinations thereof. More typically, R1, R2, R3, and R4 are each independently selected from: hydrogen, alkyl, alkenyl, aryl, or combinations thereof. Most typically, R1, R2, R3, and R4 are each independently selected from: hydrogen, alkyl, aryl, or combinations thereof. Usually, at least one of R1, R2, R3, and R4 is hydrogen; more usually, at least two of R1, R2, R3, and R4 are hydrogen; even more typically, at least three of R1, R2, R3, and R4 are hydrogen; most typically, each of R1, R2, R3, and R4 is hydrogen.

[0038] In addition, R 14 R 15 R 16 R 17 R 18 R 19 R 20 and R 21 Each is independently selected from: hydrogen, alkyl, alkenyl, aryl, halogen, hydroxyl, alkoxy, or combinations thereof. More typically, R 14 R 15 R 16 R 17 R 18 R 19 R 20 and R 21 Each is independently selected from: hydrogen, alkyl, alkenyl, aryl, or combinations thereof. Most typically, R 14 R 15 R 16 R 17 R 18 R 19 R 20 and R 21 Each is independently selected from: hydrogen, alkyl, aryl, or combinations thereof. Typically, R 14 R 15 R 16 R 17 R 18 R 19 R 20 and R 21 At least one of them is hydrogen; more generally, R 14 R 15 R 16 R 17 R 18 R 19 R 20 and R 21 At least two of them are hydrogen; more typically, R 14 R 15 R16 R 17 R 18 R 19 R 20 and R 21 At least three of them are hydrogen; even more typically, R 14 R 15 R 16 R 17 R 18 R 19 R 20 and R 21 At least four of them are hydrogen; most typically, R 14 R 15 R 16 R 17 R 18 R 19 R 20 and R 21 At least five of them are hydrogen. Typically, R... 14 R 15 R 16 R 17 R 18 R 19 R 20 and R 21 Each is hydrogen. Understandably, if X3 or X4 is a heteroatom (e.g., nitrogen or oxygen), it's possible that R is omitted respectively. 17 and R 18 .

[0039] As those skilled in the art will understand, the three substituents associated with each of R1, R2, R3 and R4 (e.g., R4', R4" and R4"' for R4) need not be the same.

[0040] Typically, R a Selected from: hydrogen, alkyl, alkenyl, aryl, halogen, hydroxyl, alkoxy, carboxylic acid, amino, amide, nitro, or combinations thereof. Typically, R a Selected from: hydrogen, alkyl, aryl, halogen, hydroxyl or alkoxy; more typically hydrogen, alkyl or alkoxy; even more typically hydrogen or alkyl; and most typically R a It is hydrogen.

[0041] The inventors have discovered that compounds of formulas (I) and (Ia) using bidentate ligands (A1 to A4) exhibit remarkable efficacy against a wide variety of microorganisms. In particular, while there are no particular restrictions on the choice of bidentate ligands, compounds according to formula (III) have been found to be especially effective. Such bidentate ligands are shown below:

[0042]

[0043] in,

[0044] Z1 and Z2 are each independently selected from: N, O, S; and

[0045] R5, R6 and R7 are each independently selected from: hydrogen, alkyl, alkenyl, aryl, halogen, haloalkyl, haloalkenyl, haloaryl, hydroxyl, alkoxy, carboxylic acid, amino, amide, nitro or a combination thereof.

[0046] Without being bound by theory, it is believed that compounds with the general structure of formula (III) containing two donor atoms at positions 4 and 5 produce compounds with remarkable antimicrobial efficacy. Although the individual donor atoms Z1 and Z2 can differ, they are typically identical. Furthermore, each of Z1 and Z2 can be independently selected from N or O. Most typically, at least one of Z1 and Z2 is nitrogen, and usually each of Z1 and Z2 is nitrogen. In this regard, it is typical that A1, A2, A3, and A4 are each independently phenanthroline or a derivative thereof.

[0047] Each of the substituents R5, R6, and R7 represents those groups attached to the ring constituting the general structure of formula (III). R5 and R7 involve up to three substituents, and R6 involves up to two substituents, as shown in formula (IIIa):

[0048]

[0049] R5, R6, and R7 may be the same or different; and each of their substituents may be the same or different. For example, when R5 is an alkyl group, one, two, or three of R5', R5"', and R5"' may be alkyl groups. Typically, R5, R6, and R7 are each independently selected from: hydrogen, alkyl, alkoxy, aryl, or combinations thereof. Typically, at least one of R5, R6, and R7 is hydrogen; typically, at least two of R5, R6, and R7 are hydrogen; in some cases, each of R5, R6, and R7 is hydrogen.

[0050] In an alternative embodiment, typically at least one of R5, R6, and R7 is an alkyl group; typically, at least two of R5, R6, and R7 are alkyl groups; in some cases, each of R5, R6, and R7 is an alkyl group. In some cases, two of R5, R6, and R7 are alkyl groups. Typically, when two of R5, R6, and R7 are alkyl groups, R5 and R7 are alkyl groups. Typical alkyl groups are as defined above. However, typically, when R5, R6, or R7 is an alkyl group, the alkyl group is selected from methyl, ethyl, or propyl. Typically, the alkyl group is methyl or ethyl, and more typically, the alkyl group is methyl.

[0051] In another alternative embodiment, typically at least one of R5, R6, and R7 is aryl; typically, at least two of R5, R6, and R7 are aryl; in some cases, each of R5, R6, and R7 is aryl. In some cases, two of R5, R6, and R7 are aryl. Typically, when two of R5, R6, and R7 are aryl, R5 and R7 are aryl. Typical aryl groups are as defined above. However, typically, when R5, R6, or R7 is aryl, the aryl group is selected from phenyl, pyridyl, or furanyl. Typically, the aryl group is phenyl or pyridyl, and more typically, the aryl group is phenyl.

[0052] It is possible that R5 and R7 are the same. Furthermore, the bidentate ligand is typically symmetrical, meaning that the substitution pattern of the structure of formula (IIIa) is symmetrical, for example, where R5' and R7' are methyl groups and all other substituents are hydrogen.

[0053] Typical bidentate ligands are compounds (1) to (7) shown below:

[0054]

[0055]

[0056] Typically, at least two of A1, A2, A3, and A4 are identical. Typically, at least three of A1, A2, A3, and A4 are identical; and it is possible that all four of A1, A2, A3, and A4 are identical. In some embodiments, each of A1, A2, A3, and A4 is different. Each of A1, A2, A3, and A4 may be independently selected from compounds (1) through (7). Typically, A1, A2, A3, and A4 are each independently selected from compounds (1), (2), (3), (5), (6), and (7); more typically, A1, A2, A3, and A4 are each independently selected from compounds (1), (2), and (3); even more typically, A1, A2, A3, and A4 are each independently selected from compounds (2) and (3); most typically, A1, A2, A3, and A4 are each independently selected from compounds (1) and (2). It is possible that A1, A2, A3 and A4 are all represented by compound (3).

[0057] In the case of compounds of formula (IIa), each of A1 and A2 can be independently selected from compounds (1) through (7). Typically, A1 and A2 are each independently selected from compounds (1), (2), (3), (5), (6), and (7); more typically, A1 and A2 are each independently selected from compounds (1), (2), and (3); even more typically, A1 and A2 are each independently selected from compounds (2) and (3); and most typically, A1 and A2 are each independently selected from compounds (1) and (2). It is possible that both A1 and A2 are represented by compound (3).

[0058] Typically, the compounds of the first aspect of this invention are positively charged, generally 4+ for compounds of formula (I) and 2+ for compounds of formula (Ia). A variety of counterions can be provided to balance this charge. While there are no particular limitations on the choice of counterions, they are typically selected from: chlorides, fluorides, bromides, hydroxides, nitrates, hexafluorophosphates, or combinations thereof. Typically, the counterions are chlorides or nitrates, usually chlorides. For the avoidance of doubt, the description of compounds herein is also intended to include descriptions of their pharmaceutically acceptable salts.

[0059] In a second aspect of the invention, a composition comprising a compound according to the first aspect of the invention is also provided. There are no particular limitations on the content of the composition. The composition may contain one or more excipients to modify the physical or chemical properties of the composition, as is well known to those skilled in the art. The composition may be formulated as a tablet for oral administration, a topical formulation suitable for human use, or a composition suitable for intravenous or subcutaneous administration, as is well known to those skilled in the art.

[0060] The composition may contain one or more additional active pharmaceutical ingredients, including one or more additional antimicrobial agents, typically one or more additional antibiotics. The composition may also be prepared using a dosage range of compounds from the first aspect of the invention.

[0061] The compounds according to the first aspect of the invention and the compositions according to the second aspect of the invention can be used to treat one or more diseases or conditions caused by microorganisms. More typically, the compounds according to the first aspect of the invention and the compositions according to the second aspect of the invention are used to treat one or more diseases or conditions caused by bacteria, particularly Gram-negative bacteria. Typical diseases include, but are not limited to: pneumonia, tuberculosis, cholera, syphilis, typhoid fever, tetanus, nosocomial infections (iatrogenic), urinary tract infections, bloodstream infections, or combinations thereof.

[0062] In a third aspect of the invention, a method for treating a microbial disease or condition is also provided, comprising the step of administering to a patient a compound of the first aspect of the invention or a composition of the second aspect of the invention. Typically, the microbial disease or condition is a bacterial disease or condition usually caused by Gram-negative bacteria. Typical examples include, but are not limited to: pneumonia, tuberculosis, cholera, syphilis, typhoid fever, tetanus, nosocomial infections (iatrogenic), urinary tract infections, bloodstream infections, or combinations thereof.

[0063] In a fourth aspect of the invention, a compound according to formula (IV) or (IVa) is provided:

[0064]

[0065]

[0066] in

[0067] R8, R8', R8”, R8”’, R9, R9’, R9”, R9”’, R 10 R 10 '、R 10 "、R 10 ”'、R 11 R 11 '、R 11 "、R 11 ”'、R 12 R 12 '、R 12 "、R 12 ”'、R 13 R 13 '、R 13 "and R 13 Each is independently selected from: hydrogen, alkyl, alkoxy, alkenyl, and aryl;

[0068] The conditions are R8, R8', R8”, R8”', R9, R9', R9”, R9”', R 10 R 10 '、R 10 "、R 10 ”'、R 11 R 11 '、R 11 "、R 11 ”'、R 12 R 12 '、R 12 "、R 12 ”'、R 13 R 13 '、R 13 "and R 13At least one of the following is selected from: alkyl, alkoxy, alkenyl, and aryl (wherein each of the alkyl, alkoxy, alkenyl, and aryl groups is as described above); and wherein R 14 To R 16 and R 18 To R 21 As stated above.

[0069] Typically, two, three, or all four of R8, R8', R8" and R8"' are the same. Similarly, typically two, three, or all four of R9, R9', R9" and R9"' are the same. In some implementations, R... 10 R 10 '、R 10 "and R 10 Two, three, or all four characters in the '' can be the same, and typically, R 11 R 11 '、R 11 "and R 11 Two, three, or all four characters in ' can be the same.' Furthermore, R... 12 R 12 '、R 12 "and R 12 Two, three, or all four in the ' are typically the same, and usually, R 13 R 13 '、R 13 "and R 13 Two, three, or all four in the '' are identical. It will be understood that it is generally easier to create complexes using the same bidentate ligands (ideally all four bidentate ligands are identical) than using combinations of different bidentate ligands, because it is difficult to reliably and precisely control the substitution pattern around the ruthenium metal center. Typically, separation techniques are required to achieve complexes with consistent "mixed" ligands.

[0070] Similarly, for equation (IVa), R8 and R8' are usually the same. And R9 and R9' are usually the same. In some implementations, R... 10 and R 10 'Can be the same, and typically R 11 and R 11 'It's the same.' Furthermore, R 12 and R 12 'Typically they are the same, and usually R 13 and R 13 'They are the same.'

[0071] In one embodiment of the present invention, R8 to R 13 At least one of them is an alkyl group; more typically, R8 to R13 At least two of them are alkyl groups; or even more typically, R8 to R 13 At least three of them are alkyl groups; more typically, R8 to R 13 At least four of them are alkyl groups; even more typically, R8 to R9 are alkyl groups. 13 At least five of them are alkyl groups; most typically, R8 to R9 are alkyl groups. 13 Each of these is an alkyl group. The alkyl group is typically selected from methyl, ethyl, and propyl, most typically methyl or ethyl, and usually methyl. Substituents R8 to R 13 Those that are not alkyl groups are typically hydrogen. Generally, the substitution around bidentate ligands is symmetrical (e.g., where R8, R9, and R...). 10 respectively with R 13 R 12 and R 11 (Same). Typically, R8 to R 13 Two of them are alkyl groups, more typically R8 to R 13 Four of them are alkyl groups; in some cases, R8 to R9 are alkyl groups. 13 All six of them are alkyl groups.

[0072] Normally, R8 and R 13 It is an alkyl group (usually methyl); typically, in this case, R9 to R 12 It is hydrogen. Or, possibly, R9 to R 12 It is an alkyl group (usually methyl); typically, in this case, R8 and R 13 It is hydrogen. Additionally, R9 and R... 12 It is an alkyl group (usually methyl); typically, in this case, R8, R 10 R 11 and R 13 It is hydrogen. Or, possibly R 10 and R 11 It is an alkyl group (usually methyl); typically, in this case, R8, R9, R 12 and R 13 It is hydrogen.

[0073] In another embodiment of the invention, R8 to R 13 At least one of them is aryl; and more typically, R8 to R 13 At least two of them are aryl groups. The aryl groups are typically selected from phenyl, pyridyl, and furanyl; most typically phenyl. Substituents R8 to R 13 Those that are not aryl are typically hydrogen. Usually, the substitution around the bidentate ligand is symmetrical (e.g., where R8, R9, and R...). 10 respectively with R 13 R12 and R 11 (Same). Typically, R8 to R 13 Two of them are aryl, and in some cases, R8 to R 13 Four of them are aryl groups.

[0074] Normally, R 10 and R 11 It is aryl (usually phenyl); typically, in this case, R8, R9, R 12 and R 13 It is hydrogen. Or, possibly, R9 to R 12 It is aryl (usually phenyl); typically, in this case, R8, R 10 R 11 and R 13 It is hydrogen.

[0075] It is possible that the compound or a pharmaceutically acceptable salt thereof is represented by formula (Va) or formula (Va):

[0076]

[0077] There are no particular restrictions on the selection of counterions that can be used; exemplary counterions are as described above.

[0078] In a fifth aspect of the invention, a composition comprising a compound according to a fourth aspect of the invention is also provided. There are no particular limitations on the content of the composition. The composition may contain one or more excipients to modify the physical or chemical properties of the composition, as is well known to those skilled in the art. The composition may be formulated as a tablet for oral administration, a topical formulation suitable for human use, or a composition suitable for intravenous or subcutaneous administration, as is well known to those skilled in the art.

[0079] The composition may contain one or more additional active pharmaceutical ingredients, including one or more antimicrobial agents, typically one or more antibiotics. The composition may also be prepared using compounds of the first aspect of the invention within a specified dosage range.

[0080] The compounds according to the fourth aspect of the invention and the compositions according to the fifth aspect of the invention can be used to treat one or more diseases or conditions caused by microorganisms. More typically, the compounds according to the fourth aspect of the invention and the compositions according to the fifth aspect of the invention are used to treat one or more diseases or conditions caused by bacteria, particularly Gram-negative bacteria. Typical diseases include, but are not limited to: pneumonia, tuberculosis, cholera, syphilis, typhoid fever, tetanus, nosocomial infections (iatrogenic), urinary tract infections, bloodstream infections, or combinations thereof.

[0081] In a sixth aspect of the invention, a method for treating a microbial disease or condition is also provided, comprising the step of administering to a patient a compound of the fourth aspect of the invention or a composition of the fifth aspect of the invention. Typically, the microbial disease or condition is a bacterial disease or condition usually caused by Gram-negative bacteria. Typical examples include, but are not limited to: pneumonia, tuberculosis, cholera, syphilis, typhoid fever, tetanus, nosocomial infections (iatrogenic), urinary tract infections, bloodstream infections, or combinations thereof.

[0082] The invention will now be described with reference to the accompanying drawings, solely for purposes of aiding understanding. Attached Figure Description

[0083] Figure 1 A shows 4 4+ The effects of uptake and cell death. Complex 4 4+ In vitro, dose-dependent killing of *Escherichia coli* MG1655 (top) and EC958 (bottom) planktonic cultures was induced. In GDMM, doses below and above 4... 4+ Different concentrations of the complex were added at MICs (1.2 μM (A) or 1.6 μM (B)). Killing was determined by monitoring the number of colony-forming units (CFU) per milliliter at time intervals of up to 6 hours post-treatment. Error bars represent three independent biological replicates ± standard deviation (SD).

[0084] Figure 1 B shows 4 4+ Effects of glucose uptake on cell death. E. coli EC958 exposed to 4 mg of glucose in the absence of glucose (left) and in the presence of glucose (right). 4+ ICP-AES data for ruthenium uptake were then analyzed. Ru (upper line) and Fe (lower line) levels per cell were expressed as metal (g) per cell. Fe levels were calculated as a control. Condition: 4 4+ The concentration was 0.8 μM. Cells were washed with 0.5% (v / v) nitric acid to remove unbound complexes. Error bars represent three independent biological replicates ± SD.

[0085] Figure 2 The image shows four nanoscopes visualized at 5, 20, 60, and 120 minutes using laser scanning confocal microscopy (LSCM) and stimulated emission loss (STED) nanomirrors. 4+ Localization in *E. coli* EC958 cells. Top row: Localization using a white laser and a 470nm notch filter at 470nm excitation. 4+Cells imaged using emission. Middle row: Cells imaged using the same excitation and emission settings, employing a 775nm depletion laser and a 780nm vortex phase plate to obtain the STED effect. Deconvolution diffraction-limited (d-LSCM) and super-resolution (d-STED) images were both processed using commercial Huygens software (SVI). Arrows highlight 4. 4+ Regions of preferential accumulation. Bottom row: Normalized emission intensity status of white solid lines drawn along selected regions of cells shown in the middle row; black solid lines represent d-LSCM, and red solid lines represent d-STED. Conditions: with 0.8 μM 4 4+ After treatment, the cells were washed with nitric acid and then fixed with paraformaldehyde (16%).

[0086] Figure 3 Representative images of cell profiles and super-resolution images (d-LSCM and d-3D STED) at the full-volume deconvolution diffraction limit are displayed. Normalized emission intensity patterns are plotted for the white solid lines drawn over the selected cell regions. Black solid lines represent d-LSCM, and red solid lines represent d-3D STED. Enlarged areas of the white dashed squares plotted at each time point, along with orthogonal representations of each axis (XY, XZ, and YZ), are shown near the images, where 4 4+ Improved resolution and better localization are shown in green. The 10 and 20-minute time points show accumulation at specific locations within the cell membrane; by the 60-minute time point, the compound is localized to portions of the cell poles, as seen in the magnified areas (a, b, c, d). The conditions used are consistent with... Figure 2 The same conditions are used in the middle.

[0087] Figure 4 A shows evidence of membrane damage in E. coli EC958. Visualized by SIM at 5 minutes (left) and 60 minutes (right). 4+ Co-localization with NHS-ester405. (i) Using 0.8 μM 4 4+ Cells were treated and fixed with paraformaldehyde (16%). After fixation, cells were treated with 2.5 μg / mL NHS-ester405. Top image: 4 4+ Emissions (A568 filter); Middle image: Emissions of NHS-ester405 at 405nm (DAPI filter); Bottom image: Combined image. (ii) In the absence of 4 4+ In the case of staining with NHS-ester, the staining is performed.

[0088] Figure 4 B showed 4 4+Induced ATP release from EC958 cells, extracellular [ATP] (nM) was quantified using recombinant luciferase and D-luciferin, with ATP release measured using a spectrophotometer. Samples were exposed to 0 (control), 0.8, and 1.6 μM (MIC) at 4... 4+ Two hours. A significant difference in the three-star result was observed between 0 and 1 MIC, P = 0.0006. Error bars represent three independent biological replicates ± SD. The ATP positive control was polymyxin 4 μg / mL (inset). IC50 against HEK293 cells and three bacterial strains. 50 / MIC comparison.

[0089] Figure 4 C shows the IC50 values ​​for HEK293 cells and three bacterial strains. 50 / MIC comparison.

[0090] Figure 4 D shows the Kaplan-Meier survival curves for toxicity screening of wax moths, where cells were treated with 0-80 mg / kg of compound 4. 4+ Treatment, cultured at 37.5°C for 120 hours: water control (black), compound 4 4+ (red).

[0091] Figure 5A The UV-Vis absorption spectra are shown, illustrating the change in molar extinction coefficient with increasing concentration of [{Ru(3,4,7,8-tetramethyl-1,10-phenanthroline)2}2(tpphz)] in MeCN (performed using a Cary 300 UV / Vis spectrophotometer at 27.5 °C).

[0092] Figure 5B The UV-Vis absorption spectra are shown, illustrating the change in molar extinction coefficient with increasing concentration of [{Ru(3,4,7,8-tetramethyl-1,10-phenanthroline)2}2(tpphz)] in water (performed using a Cary 300 UV / Vis spectrophotometer at 27.5 °C).

[0093] Figure 6A The emission spectra in MeCN at increasing concentrations of [{Ru(3,4,7,8-tetramethyl-1,10-phenanthroline)2}2(tpphz)] are shown in the range of 550–800 nm. The fluorescence spectra were obtained using a Fluoromax 3 fluorometer at 27.5 °C.

[0094] Figure 6BThe emission spectra of [{Ru(3,4,7,8-tetramethyl-1,10-phenanthroline)2}2(tpphz)] in water are shown in the range of 550–800 nm. The measurements were performed using a Fluoromax 3 fluorometer at 27.5 °C.

[0095] Figure 7 The Log P values ​​and activity (MIC) of each complex against pathogenic Escherichia coli strain EC958 are compared to demonstrate that activity increases with relative lipophilicity. Log P data were collected using a shake-flask procedure. “Phen” stands for phenanthroline, “DMP” for dimethylphenanthroline, “TMP” for tetramethylphenanthroline, and “DIP” for diphenylphenanthroline.

[0096] Figure 8 The EC958 was shown to be 4 4+ Accumulate CFU / mL counts to ensure that the number of bacteria in the solution remains constant between each time point.

[0097] Figure 9 The results show that at each time point, under the conditions of (+) glucose presence and (-) glucose absence, for 4... 4+ Regarding the differences in Ru content per cell (g), significant differences were observed in ruthenium accumulation in samples with (+) glucose at 10 and 20 minutes. Ruthenium content per cell was determined by ICP-AES.

[0098] Figure 10A The results showed that using Fluoromax 3, in Tris buffer at 27°C, with increased concentrations of CT-DNA and 4 4+ DNA binding titration.

[0099] Figure 10B The Scatchard plot and McGhee Von Hippel fit are shown to obtain 4 4+ The binding constant.

[0100] Figure 11A and 11B This describes the detection of membrane potential in *E. coli* EC958 cells. The percentage (%) of cell populations showing red / red+green / green fluorescence is given. The presence or absence of 1.6 μM 4 4+ Under these conditions, cells were cultured with 30 μM Di-OC2(3) for 30 minutes.

[0101] Figure 12A and 12B The analysis of flow cytometry data using red and green fluorescence parameters is shown. Dot plots of red fluorescence versus green fluorescence were collected under logarithmic magnification.

[0102] Figure 13 Showing the use of 4 4+ Acinetobacter baumannii (AB184) stained at MIC concentration for 10 min (left) and 60 min (right), corresponding to the outer membrane (10 min) and inner membrane (60 min) imaging. Cells were fixed with PFA (4%) and washed with PBS. Cells were imaged using a structured illumination microscope with a 488 nm laser and an A568 filter.

[0103] Figure 14 The figures show the ruthenium hemolymph content (μg / mL) of wax moths injected at 20 mg / kg (blue) and 80 mg / kg (red). 10 μL of 4 4+ Water was injected into the left ventral leg of the wax moth, and the moths were incubated at 37.5°C for 120 hours. The moths were scored at each time interval: survival / death, activity, and melanization. Ru content was determined by ICP-AES.

[0104] Figure 15 Displaying wax moth (CFU 10) 5 left, 10 6 Right) Toxicity screening Kaplan-Meier survival curves, cells were used at 0-80 mg / kg in 4... 4+ Treatment was performed and incubated at 37.5°C for 120 hours; water control (orange), bacteria (green), and 4... 4+ (Purple). The larvae were injected with bacteria into their right gastropod, followed by 4 gram of the bacteria into their left gastropod 30 minutes later. 4+ .

[0105] Figure 16 A shows the 4 at 40 mg / kg and 80 mg / kg 4+ Bacterial CFU counts from extracted larval hemolymph (wax moth) were performed after 120 hours of observation in the presence of the bacteria. Extraction was conducted at 24 and 120 hours. Initial bacterial count was 10. 5 (left) and 10 6 (Right). Larvae were cultured at 37.5°C.

[0106] Figure 16 B shows a photograph of the agar plate, which was used to determine... Figure 16 Figure A shows the results after 24 hours at doses of 0 mg / kg, 40 mg / kg, and 80 mg / kg. 4+ Complete clearance was observed within 96 hours at single doses of 40 and 80 mg / kg. The black markings were not bacterial colonies, but rather blackened hemolymph.

[0107] Figure 17 Four images were observed using a confocal microscope. 4+Localization of Acinetobacter baumannii AB184 cells in larval hemolymph. Top row: Excited at 450 nm using an A568 filter with 4... 4+ Cells in emission imaging. Middle row: phase contrast. Bottom row: composite image. Extracted hemolymphocytes were washed with nitric acid and then fixed with paraformaldehyde (16%).

[0108] Figure 18 AC is an image obtained by removing wax moth larvae. Complex 4 in stained Acinetobacter baumannii AB184. 4+ The luminescence of the bacteria was measured (head A, tail B). Blood cells containing bacterial cell regions I and II were highlighted. Acinetobacter baumannii was analyzed using 1.2 μM 4... 4+ Stain for 30 minutes, then fix with PFA (4%). Inject larvae into cells and incubate for 30 minutes. Larvae anesthetized with ether are incubated in PFA (4%) for 30 minutes, then removed using the CUBIC protocol. Images are captured using a Nikon confocal microscope with a 488nm laser and a red emission filter. Images are processed using ImageJ.

[0109] Figure 19 A series of images of cleared wax moth larvae infected with Acinetobacter baumannii AB184 are provided, using 0.5 mg / mL NHS-ester488(A) or 1.2 μM complex 4. 4+ (B) Staining was performed. Cell regions were identified (i) and enlarged (AII, AIII, BII) to show Acinetobacter baumannii within blood cells. Three-dimensional surface maps (AIV, BIII) were presented to show peak emission intensity. Conditions and Figure 18 Same as described above. Images were captured using a fluorescence stereomicroscope: 490nm ex / 520nm em (NHS-ester488), 565nm ex / 640nm. Images were processed using ImageJ.

[0110] Figure 20 The results showed that Escherichia coli ST131 and EC958, after being subjected to different concentrations of 4 4+ Biofilm formation under the influence of ruthenium. N≥3±SEM. Biofilms formed in 24-well plates after incubation at 37°C for 19 hours. ST131 showed biofilm formation in the absence of ruthenium and at 0.156 μM (10% of the MIC).

[0111] Figure 21 This shows that when existing biofilms are exposed to 15.6 μM, 78 μM, and 156 μM of 4 4+ At the time of influence, the biofilm produced by *E. coli* ST131 showed CFU / mL. N≥3±SEM. The biofilm was grown on the filter membrane at 37°C for 4 days, with MHA replaced daily. On the fourth day, compound 4 was directly added.2+ Transferred to a biofilm and incubated for 24 hours. CFU / ml was calculated using the Miles and Mirsa methods. One-way ANOVA was performed using Turkey's multiple comparison test.

[0112] Figure 22A and 22B 4 were displayed 2+ Helical DNA binding was observed in EC958, and mononuclear TMPs again showed DNA binding in SH1000.

[0113] Figure 23 4 were displayed 2+ Cell death effects of uptake. Escherichia coli EC958 exposed to 4 mg of glucose in the absence of glucose (a) and in the presence of glucose (b) 2+ ICP-AES data for ruthenium uptake were then analyzed. Ru (upper line) and Fe (lower line) levels per cell were expressed as metal (g) per cell. Fe levels were calculated as a control. Condition: 4 2+ The concentration was 0.8 μM. Cells were washed with 0.5% (v / v) nitric acid to remove unbound complexes. Error bars represent three independent biological replicates ± SD.

[0114] Figure 24 The results show that at each time point, under the conditions of (+) glucose presence and (-) glucose absence, for 4... 2+ Differences in Ru content (g) per cell were observed. At 10 minutes, ruthenium accumulation showed significant differences in (+) glucose samples. Ruthenium content per cell was determined by ICP-AES.

[0115] Figure 25 Displayed based on Figure 1 B and Figure 23 The result, 4 2+ and 4 4+ The relative rate of Ru uptake.

[0116] Figure 26 AC displays four images visualized at 60 minutes, 120 minutes, and 24 hours using laser scanning confocal microscopy (LSCM) and stimulated emission loss (STED) nanomirrors. 2+ Localization in *E. coli* EC958 cells. The localization was achieved using a white light laser and a 470nm notch filter under 470nm excitation. 2+ Emission imaging cells were processed using commercial Huygens software (SVI) to obtain deconvolution diffraction-limited (d-LSCM) and super-resolution (d-STED) images. Conditions: 0.8 μM 4 2+ After treatment, the cells were washed with nitric acid and then fixed with paraformaldehyde (16%).

[0117] Figure 27 The results of the DNA co-staining experiment are shown. Using 4... 2+ Stained Staphylococcus aureus (top and CI), Staphylococcus aureus stained with DNA and DAPI (CII), overlaid images (CIII), overlaid 3D surface map (CIV). This image shows the direct overlay of DAPI and mononuclear TMP, with calculated Pearson colocalization constant >0.9.

[0118] Figure 28 Shown in 4 2+ The results of Ames mutation assay on Escherichia coli in the presence of [a specific substance]. Using 4 [a specific method / method]... 2+ The percentage of mutagenesis was measured by treating *E. coli* cells (25% overnight culture, 75% medium) at 0x (natural mutagenesis control), 0.5x, 1.0x, and 2.0x. Bromocresol purple indicator was added, and the percentage change in color from purple to yellow was measured after 48 hours of incubation. A positive control of cells irradiated with UV light was added for comparison.

[0119] Figure 29 Shown in 4 2+ TEM images of Escherichia coli are present.

[0120] Figure 30 Showing 4 2+ Induced ATP release from *E. coli* EC958 cells, extracellular [ATP] (nM) was quantified using polymyxin and measured using a spectrophotometer. Samples were exposed to 0 MIC (control), 0.5 MIC, and 1.0 MIC for 4 days. 2+ Four hours. Error bars represent three independent biological replicates ± SD. Polymyxin concentration was 4 μg / mL.

[0121] Figure 31 Kaplan-Meier survival curves for toxicity screening of wax moths were shown, with cells treated with 0-80 mg / kg of 4... 2+ Treatment, cultured at 37.5°C for 120 hours: water control (black), compound (red).

[0122] Figure 32 Kaplan-Meier survival curves for Acinetobacter baumannii AB184 virulence screening are shown (initial bacterial count: 10). 4 left, 10 5 (Right), cells are treated with 40 or 80 mg / kg of 4 2+ The mixture was treated and incubated at 37.5°C for 120 hours: water control (red), compound (green).

[0123] Figure 33 A shows the concentrations at 40 mg / kg and 80 mg / kg.2+ After 120 hours of observation in the presence of bacteria, the bacterial CFU count (Acinetobacter baumannii AB184) from the extracted larval hemolymph was recorded. The initial bacterial count was 10. 4 (left) and 10 5 (right).

[0124] Figure 33 B shows a photograph of the agar plate, which was used to determine... Figure 33 Regarding the 80mg / kg dosage in section A, 4 2+ The image shows black markings that are not bacterial colonies, but rather blackened hemolymph. Detailed Implementation

[0125] The invention will now be described with reference to specific embodiments. These embodiments should not be construed as limiting, and are provided to enhance understanding of the invention.

[0126] Example

[0127] Synthesis of binuclear complexes

[0128] Complex 1 4+ and 2 4+ (See Scheme 1) The following procedure is used for synthesis.

[0129]

[0130] Scheme 1. Compound 1 4+ Up to 4 4+

[0131] 1,10-phenanthroline-5,6-dione (Compound 1 4+ )

[0132] 1,10-Phenanthroline (18.02 g, 100 mmol) was dissolved in 60% H₂SO₄ (125 mL). Potassium bromate (66.81 g, 400.1 mmol) was slowly added with continuous stirring to prevent the reaction from becoming too vigorous. The reaction released brown fumes of bromine gas. Once all the potassium bromate had been added, the reaction mixture was cooled to room temperature. The mixture was further cooled by adding crushed ice (100 g) and placing it in an ice bath. The solution was neutralized to pH 5-6 by dropwise addition of NaOH (20 M). The mixture became hot during neutralization and had to be carried out in an ice bath to maintain cooling. The yellow precipitate was filtered onto the sintered body and washed with water (1 L) and diethyl ether (100 mL). The product was dried under vacuum. The crude product was purified by recrystallization in water / methanol (1:50), and bright yellow crystals were collected by vacuum filtration. Mass = 16.04 g (76.31 mmol, 76.3%) of yellow solid. 1¹H NMR (CDCl₃) δ (split integral): 7.61 (dd, 2H), 8.52 (dd, 2H), 9.13 (dd, 2H). MS: m / z: 210.1 (100) [M+]. tetrapyrido[3,2-a:2',3'-c:3'-c:3",2"-h:2"',3"'-j]phenazine (TPPHZ) [Ru(N-N)2Cl2]

[0133] Ammonium acetate (15 g, 194.6 mmol), diphenylphenanthrene (2.90 g, 13.8 mmol), and sodium dithionite (300 mg, 1.72 mmol) were refluxed and boiled under nitrogen at 180 °C for 2 hours. The reaction mixture was stirred occasionally. Once the reaction was complete, the mixture was cooled to room temperature and then distilled water (20 mL) was added. The resulting yellow precipitate was collected, filtered under vacuum, and washed with water, methanol, and acetone (3 × 20 mL). The resulting crude product was ground in refluxed ethanol (100 mL) to remove impurities, filtered while hot, and dried under vacuum. Mass = (0.92 g, 2.39 mmol, 34.6%) yellow solid. The product is slightly soluble in most solvents. 1 HNMR (CDCl3) δ (split integral) 7.94 (dd, 4H), 9.41 (dd, 4H), 9.83 (dd, 4H). 1 ¹H NMR (d-TFA) δ (split integral) 8.62 (dd, 4H), 9.56 (dd, 4H), 10.52 (dd, 4H). MS; m / z (42.6%): 385.1 (100) [M+].

[0134] [Ru(1,10-phenanthroline)2Cl2]

[0135] Four compounds were synthesized by the following method, where NN represents a substituted phenanthroline auxiliary ligand. RuCl3·3H2O, NN, and LiCl were refluxed in DMF and heated for 8 hours. The reaction mixture was cooled to room temperature and acetone was added. The mixture was stored at 4°C for 16 hours. The deep purple precipitate was washed with water and ethanol and dried under vacuum.

[0136] [{Ru(N-N)2}2(tpphz)][PF6]4

[0137] RuCl3·3H2O (1.56 g, 6 mmol), LiCl (1.55 g, 36.9 mmol), 1,10-phenanthroline (2.5 g, 13.9 mmol), DMF (20 mL), and acetone (100 mL). Mass = 2.41 g (4.59 mmol, 66.1% yield). ES-MS m / z (%): 497 (70) [M-Cl] + 525(100)[M-Cl] + +CO.

[0138] [{Ru(1,10-phenanthroline)2}2(tpphz)][PF6]4

[0139] Four compounds were synthesized via the following general procedure. [Ru(NN)₂Cl₂] and (tpphz) were added to a 1:1 solution of ethanol and water. The solution was refluxed under nitrogen for 12 hours. After this, the reaction mixture was cooled to room temperature and stored at 4°C for 16 hours. The red solution was filtered, and ethanol was removed by rotary evaporation. A saturated amount of NH₄PF₆ was added; this resulted in the formation of a deep red precipitate. The precipitate was collected by filtration, washed with water, and recrystallized in acetonitrile with diethyl ether. The product was dried under vacuum and purified on an alumina column using the following solvent system: 95% MeCN, 3% dH₂O, and 2% KNO₃.

[0140] Properties of binuclear complexes

[0141] Tpphz (0.263 g, 0.68 mmol), [Ru(1,10-phenanthroline)2Cl2] (1 g, 1.89 mmol), and ethanol / water (50 mL). Mass = 1.1 g (0.58 mmol, 85.6% yield). 1 H NMR (MeCN-d) 6 δ (split integral): 7.71 (m, 8H), 7.94 (dd, 4H), 8.09 (d, 4H), 8.29 (dd, 8H), 8.33 (s, 8H), 8.69 (dd, 8H), 10.01 (dd, 4H). ES-MS; m / z (%): 799 (10) [M-2PF6] 2+ ,484(15)[M-3PF6] 3+ ,321(50)[M-4PF6] 4+ Accurate quality analysis: C 72 H 28 N 14 [ 102 Ru]2 4+ The calculated value is 321.1110. The verified value is 321.1112.

[0142] Complex 3 4+ and 4 4+ A similar method was used to synthesize the relevant methylated bidentate ligands. 3 4+ and 4 4+ MeCN consistently demonstrates the expected strong Ru-based performance. II →tpphz 3 MLCT emission was observed, centered at 670 nm and 700 nm, respectively (Figures 5 and 6). The biological properties of all four complexes were investigated using chloride salts (obtained via anion displacement).

[0143] Figure 7

[0144] The balance between lipophilic and hydrophilic properties is considered crucial for the uptake of bioactive substrates by living cells. Log P values ​​for all four complexes were determined using a shake-flask procedure via octanol-water partitioning. The results are as follows: 1 4+ =1.77,2 4+ =1.03,3 4+ =1.38,4 4+ =1.13. These data indicate that 2 4+ These are the most lipophilic complexes. Furthermore, the relative lipophilicity appears to increase with the number of methyl groups linked to the auxiliary ligands of these complexes.

[0145] The bioactivity of these compounds against wild-type K12 derivative MG1655 of *Escherichia coli* and urethral inflammatory multidrug-resistant strain EC958 ST131 was investigated. The compounds were also tested against another Gram-positive bacterium, *ESKAPE* bacteria (specifically, the pathogenic gastrointestinal strain V583 (ATCC700802) of *Enterococcus faecalis*, a major opportunistic pathogen and a primary cause of urinary tract infections). The minimum inhibitory concentrations (MICs) of these four complexes were obtained in glucose minimum limit medium (GDMM) and nutrient-rich Mueller-Hinton-II (MH-II). Figure 1 As shown in Table 1, all four complexes exhibited high activity in GDMM.

[0146] Table 1. MIC (μM) and MBC (μM) results of wild-type (MG1655) and pathogenic (EC958) strains of Escherichia coli and pathogenic (V583) strain of Enterococcus faecalis in GDMM and MH-II.

[0147]

[0148] Although compound 2 is the most lipophilic 4+ The series exhibited the lowest activity (likely due to its low solubility in aqueous media), while the lipophilic series showed increased lipophilicity accompanied by increased activity, with 4 of them showing the lowest activity. 4+ It exhibits the highest activity against all three bacterial strains. Notably, 1 4+ 3 4+ and 4 4+ It showed considerable activity against β-lactam-resistant strains of *Escherichia coli* and vancomycin-resistant strains of *Enterococcus faecalis*; Complex 4 4+ Its activity against wild-type strains of Escherichia coli is even higher than that against ampicillin. Furthermore, it showed activity against 1... 4+ Up to 4 4+The estimated minimum bactericidal concentration (MBC) values ​​are summarized in Table 1. These data indicate that, in terms of MIC data, an increase in MBC values ​​was observed between GDMM and MH-II. Similarly, 4 4+ It exhibits the highest activity, with an MBC value lower than ampicillin in the GDMM, indicating greater activity than conventional antibiotics against all bacterial strains. Furthermore, since the MBC values ​​against all three strains exceed the MIC by at least four times, all compounds act as antimicrobial agents.

[0149] In determining coordination compound 4 4+ After demonstrating the most promising bactericidal properties, time-kill kinetics were performed on two strains of *E. coli*, after exposure to an increased concentration of the complex in a minimal culture medium at 37°C for 6 hours (see [link to study]). Figure 8 A and Figure 1 At concentrations below the MIC, colony-forming units (CFU) gradually increased as bacteria continued to grow. In both strains, it was clear that at MIC and higher concentrations, the compound inhibited bacterial proliferation and reduced the number of viable bacterial cells. For exposure to the highest concentration of 4... 4+ EC958 showed no colony formation, indicating that all bacteria in the system had been killed. The difference between the MBC value and the time-kill test may stem from different experimental conditions, where the MBC test involves static incubation for 16-18 hours, while the time-kill test is performed under 90% aeration and rotation for 6 hours.

[0150] To investigate the effects of E. coli cells on 4 4+ The uptake of glucose was studied using ICP-AES. Four experiments were conducted in the presence and absence of glucose. 4+ Intake studies (see) Figure 9 B and Figure 2 Experiments were conducted to study the accumulation of ruthenium in Escherichia coli EC958 within 1 hour. At high concentrations, time-kill assays showed that 99.9% of the bacteria survived exposure to 4 hours. 4+ They were killed within the first hour. In these experiments, the concentration of iron (a trace element in all cells) was also quantified as a control. It was found that using 4... 4+ During treatment, the iron content remained constant; furthermore, the change in CFU / mL was negligible, indicating that the cells did not lyse during the accumulation experiment.

[0151] Under glucose-free conditions, accumulation exhibited two phases: after an initial increase upon initial exposure, ruthenium remained at low levels for approximately 20 minutes, followed by a gradual doubling of uptake, eventually reaching 1.1 × 10⁻⁶ per cell. -16 g. Assuming an average cell volume of 1 μm. 3This equates to an intracellular concentration >1 mM. In contrast, in the presence of glucose, although the final amount of ruthenium accumulated was the same within experimental error, the uptake of the complex was rapid, reaching maximum intracellular ruthenium concentration within 20 minutes. Significant differences between glucose- and glucose-free conditions were observed at 10 and 20 minutes.

[0152] In addition, metal complexes were used to perform super-resolution analysis on 4 4+ Uptake and cellular response to exposure to 4 4+ The response was analyzed. Structured illumination microscopy was used to improve the resolution (approximately 100 nm) for the analysis of 4... 4+ For imaging of the internalization, we also used stimulated emission loss (STED) nanomicroscopy to provide the highest sub-diffraction-limited resolution. Example STED images taken over a time course (5–120 minutes) are shown below. Figure 3 .

[0153] To investigate whether cell morphology changed during the first 5–20 minutes of exposure, images were taken at the same time points and under the same conditions used in the accumulation experiment. These images confirmed 4 4+ It is readily and rapidly taken up by pathogenic strains of E. coli. Interestingly, up to 20 minutes, the complex accumulates in large quantities at the cell membrane and is generally distributed within the cell compartments. However, after this period, it increasingly preferentially resides at the cell poles.

[0154] STED microscopy has also been used for detailed 3D partitioning experiments (3D STED). Considering the critical factor of bacterial cell size, the use of dual STED beams split into the XY plane and Z axis improves the highest possible 3D resolution for each imaging plane. Using this procedure, a 3D STED resolution of 50 nm per plane and approximately 120 nm on the Z axis was obtained. Figure 4 It showed that exposure to 4 4+ Images taken at specific time points after the initial exposure. During the first 10 minutes, the probe accumulates within the cell membrane, forming a unique distribution pattern. After 20 minutes, dye redistribution begins, with accumulation at the cell poles becoming increasingly apparent.

[0155] Combined with ICP-AES data, imaging studies showed that the uptake and intracellular distribution of the complex changed after approximately 20 minutes. Furthermore, due to 4 4+ The molecular weight of the ester is significantly greater than the upper limit of porin-mediated uptake (approximately 600 Da), therefore this mechanism can be ignored. To investigate the possibility of membrane damage, a secondary co-staining experiment was performed using the probe AlexaFluorNHS-ester405 (see [link to study]. Figure 4 A).

[0156] Because Alexa Fluor NHS-ester405 is impermeable to undamaged bacterial membranes, it can be used for cell membrane localization and imaging. When exposed to 4... 4+ Five minutes later, NHS-ester 405 localized to the bacterial cell membrane. However, after 60 minutes of exposure to the complex, both dyes were found to be internalized in *E. coli*. In contrast, even after 60 minutes, cells stained with NHS-ester 405 alone continued to show membrane staining only. Membrane staining was only observed after using 4... 4+ The fact that internalization only occurs after treatment further proves that the complex is disrupting the bacterial membrane structure. To study this phenomenon more quantitatively, a concentration-dependent ATP cell leakage assay was performed. Using a specific concentration of 4... 4+ After treatment, the presence of extracellular ATP released after bacterial cell membrane damage was detected by using luminescence generated by the ATP-dependent reaction between recombinant firefly luciferase and D-luciferin.

[0157] Data obtained from luminescence-based [ATP] assays (summarized in...) Figure 4 B) Confirmed that when exposed to 4 4+ At this time, the bacterial membrane is damaged in a concentration-dependent manner. Given this effect, the uptake of the complex appears to be biphasic under glucose-free conditions, since membrane damage must occur within the internalized 4-day timeframe. 4+ This occurs before the levels rise to high concentrations. This membrane damage may be the only mechanism of the complex's therapeutic effect, although the complex's localization and binding to specific cellular regions once internalized suggests another cellular target. Given that pathogenic, therapeutically resistant *E. coli* strains remain sensitive to this complex, it appears that 4 4+ The membrane disruption effect may be just one aspect of a more complex series of interactions and cellular responses.

[0158] Table 2. UV-Vis spectral data, showing the molar extinction coefficients and maximum absorbance of four [{Ru(NN)2}2(tpphz)] in water and MeCN, measured at 27.5 °C using a Cary 300 UV / Vis spectrophotometer.

[0159]

[0160] Table 3. Emission data, showing the molar extinction coefficients and maximum absorption values ​​of four [{Ru(NN)2}2(tpphz)] in water and MeCN, measured at 27.5 °C using a Fluoromax 3 fluorometer.

[0161]

[0162] Due to 4 4+Exhibiting high antimicrobial activity and targeting the membrane, the compound's potency in non-cancerous eukaryotic cells was determined, allowing for further exploration of its potential as a lead inhibitor for antimicrobial therapy. MTT assays in the human embryonic kidney cell line HEK293 showed a mean IC50... 50 A value of 135 μM indicates an inhibitory concentration at least 80-fold different between bacteria and HEK293 cells (see [link to relevant documentation]). Figure 4 C).

[0163] Given IC 50 Promising comparisons were made between the MIC values ​​and animal model screening. Because many aspects of the physiology of the wax moth (Galleria mellonella) larvae, particularly their immune system, are very similar to those of mammals, they have been extensively used as in vivo models, including for toxicity screening, producing results comparable to commonly used mammalian models. Using 4 4+ Toxicity screening was performed, and Kaplan-Meier survival curves were plotted (see...). Figure 13 D). All concentrations used were above 4%. 4 + The MIC for EC958 was determined, and within the daily dose range for clinical use of antimicrobial agents. The results were determined by a log-rank test at all compound concentrations, using 4... 4+ There was no significant difference in the survival percentage between the treated wax moths and the control group. Furthermore, activity and melanization scores indicated that exposure to 4... 4+ The compound showed no significant negative effects on wax moths, confirming its non-toxicity at concentrations far above the MIC.

[0164] Table 4. DNA binding constants and site sizes of the four compounds described in this paper (Figure 10)

[0165]

[0166] Table 5. Log-rank (Mantel-Cox) test of Kaplan-Meier survival curves to determine whether a significant difference was observed between the survival percentage of wax moths injected with the compound (water) and those injected with 10 μL of the compound (0-80 mg / kg) and stored at 37.5°C for 120 hours.

[0167]

[0168] 4 4+ The membrane potential and flow cytometry behavior were measured, as shown in Figures 11 and 12. Furthermore, it has been shown that 4 4+ It can penetrate Acinetobacter baumannii (AB184) at the MIC concentration. Figure 17 At 10 minutes, the outer membrane can be imaged, and at 60 minutes, the inner membrane can be imaged.Figure 14 4 were displayed 4+ Localization of Acinetobacter baumannii AB184 cells in larval hemolymph.

[0169] For injections, there are 4 4+ The ruthenium hemolymph content (μg / mL) of the wax moth was determined. Figure 15 As expected, higher doses resulted in higher ruthenium levels in the hemolymph, although these levels remained largely constant throughout the experiment. The Kaplan-Meier infection model showed higher levels of ruthenium compared to the control (water). Figure 16 ) in 4 4+ Good survival was observed at both concentrations (40 mg / kg and 80 mg / kg). Furthermore, Figure 18 Figures A and 16B show the bacterial CFU counts (caecilomyces cerevisiae) from extracted larval hemolymph. From these figures, it is clear that at both treatment concentrations (40 or 80 mg / kg), the CFU counts at different concentrations are significantly different. 4+ After 48 hours, all colonies were eliminated. This elimination was achieved with a single compound dose. In contrast, in the absence of 4... 4+ Under these conditions, bacterial growth was observed to be exponential.

[0170] Using the CUBIC removal program, wax moth larvae were selected for further study. Figure 19 Images are shown taken through the removed wax moth larvae. At the head (A) and tail (B) of the wax moth, 4 4+ Acinetobacter baumannii cells fluoresce after staining. The areas containing Acinetobacter baumannii cells are highlighted (I, II). The initiation of an immune response in *Eriocheir sinensis* hemolymphocytes is observed (C). Images show that, with 4... 4+ The stained Acinetobacter baumannii was ingested and phagocytosed by the larval hemocytes. This indicates that 4 4+ It not only kills bacteria, but also upregulates the larvae's immune response, enabling the infection to be cleared. Figure 20 Images of the removed larvae, infected with Acinetobacter baumannii AB184, are also shown. This is confirmed by NHS-ester488(A) or 4... 4+ (B) Staining. Cellular regions were identified (i) and enlarged (AII, AIII, BII) to show Acinetobacter baumannii within blood cells. Three-dimensional surface maps (AIV, BIII) were presented to show peak emission intensities.

[0171] Figure 21 The study demonstrated the effects of Escherichia coli ST131 and EC958 on complex 4. 4+ Research on biofilm formation under the influence of ST131. 4+Biofilm formation occurred at 10% of the MIC. Welch's t-test showed no significant difference between these groups (P = 0.687). At concentrations of 0.390 and 0.781, no biofilm formation was observed (with small negative values ​​of 0.003 and 0.005, respectively). Therefore, 4 4+ It can prevent biofilm formation at concentrations as low as 0.390 μM. Synthesis of mononuclear complexes This study demonstrates the effects of pre-formed biofilms. The biofilms were subjected to four [units of molecular weight] at 15.6 μM, 78 μM, and 156 μM. 4+ Treatment. A significant difference in one star was observed between the control (0 μM) and the 156 μM sample (p = 0.0015), and a significant difference in two stars was observed between the control (0 μM) and the 15.6 μM sample (p = 0.0086). Therefore, 4 4+ It has the ability to prevent the formation of biofilms and to penetrate existing biofilms.

[0172] [Ru(N-N)2Cl2]

[0173] Complex 1 2+ and 2 2+ (See Scheme 2) The following procedure is used for synthesis.

[0174]

[0175] Scheme 2. Compound 1 4+ Up to 4 4+

[0176] [Ru(3,4,7,8-tetramethyl-1,10-phenanthroline)2Cl2]

[0177] Four compounds were synthesized by the following method, where NN represents a substituted phenanthroline auxiliary ligand. RuCl3·3H2O, NN, and LiCl were refluxed in DMF and heated for 8 hours. The reaction mixture was cooled to room temperature and acetone was added. This was stored at 4°C for 16 hours. The deep purple precipitate was washed with water and ethanol and dried under vacuum.

[0178] [Ru(N-N)2(DPQ)][PF6]2

[0179] RuCl3·3H2O (1.14 g, 5.50 mmol), TMP (2.4 g, 10.16 mmol), LiCl (1.47 g, 34.68 mmol), DMF (19 mL), and acetone (100 mL). Mass = 2.07 g (3.21 mmol, 63.2%) purple solid. MS m / z (%): 609.1 (62) [M-Cl] + , 637.1(100)[M] + 667.1. (44)[M+Na] +Carbon monoxide replaces one of the chlorides.

[0180] [Ru(3,4,7,8-tetramethyl-1,10-phenanthroline)(DPQ)][PF6]2

[0181] Four compounds were synthesized via the following general procedure. [Ru(NN)₂Cl₂] and DPQ were suspended in a 1:1 solution of EtOH:H₂O. The suspension was refluxed under argon for 12 hours, cooled to room temperature, and filtered. NH₄PF₆ was added to form a brown hexafluorophosphate.

[0182] [Ru(N-N)2(tpphz)][PF6]2

[0183] [Ru(TMP)₂Cl₂] (1.01 g, 1.57 mmol), DPQ (0.495 g, 2.36 mmol), and EtOH:H₂O (50 mL). Mass = 0.861 g (0.801 mmol, 51%). MS (TOF MS LD⁺) m / z (%): 784 (51) [M⁻²PF₆) 2+ ,929(100)[M-PF6] + 1H NMR (DMSO-d) 6 δ(split integral): 2.23(6H,s), 2.39(6H,s), 2.79(6H,s), 2.85(6H,s), 7.46(2H,dd), 7.85(2H,d), 7.65(2H,s), 7.95(2H,s), 8.41(4H,d), 8.48(2H,d).

[0184] [Ru(3,4,7,8-tetramethyl-1,10-phenanthroline)(tpphz)][PF6]2

[0185] Four compounds were synthesized via the following general procedure. 5,6-Diamino-1,10-phenanthroline was dissolved in hot methanol, and then [Ru(NN)₂DPQ][PF₆]₂ was added to a boiling solution in acetonitrile. The reaction mixture was refluxed at 80 °C for 6 hours. The solution was cooled to room temperature and filtered. NH₄PF₆ was added to form a red hexafluorophosphate. The crude product was washed with water, ethanol, and diethyl ether. It was then purified on a primary alumina column using acetonitrile / water / KNO₃. The red range was collected, the solvent was removed under reduced pressure, and the red solid was dried under vacuum.

[0186] Properties of mononuclear complexes

[0187] 5,6-Diamino-1,10-phenanthroline (88.2 mg, 0.42 mmol), hot methanol (17 mL), [Ru(TMP)2DPQ][PF6]2 (606 mg, 0.56 mmol), acetonitrile (30 mL). Mass = 0.272 g (0.389 mmol, 45%), 1H NMR (CD3CN-d 6 )δ (split integral): 2.29(6H,s),2.32(6H,s),2.80(6H,s),2.86(6H,s),7.71-7.78(4H,m),7.84(4H,s),8.15(4H,d),9.30(4H,d),9.59(4H,d). MS; m / z(%): 479[M-2(PF6)] 2+ Accurate quality analysis: C 56 H 44 N 10 [ 102 Ru] 2+ The calculated value is 479.1391. The verified value is 479.1405.

[0188] Figure 23

[0189] Using the same compound 1 determined above 4+ Up to 4 4+ Under the same parameters and conditions as the MIC and MBC values, complex 1 was tested. 2 + Up to 4 2+ The MIC and MBC values.

[0190] Table 6. MIC results of mononuclear complexes

[0191]

[0192]

[0193] Table 7. MBC results of mononuclear complexes

[0194]

[0195] As can be seen from Tables 6 and 7, compound 1 2+ Up to 4 2+ Each of them was found to have antimicrobial activity. Furthermore, compound 4 was found... 2+ It has shown remarkable efficacy as an antibiotic, and its properties are superior to existing antibiotics (such as ampicillin). To avoid confusion: SH1000 is Staphylococcus aureus; AB184 is Acinetobacter baumannii; PA2017 is Pseudomonas aeruginosa.

[0196] In determining coordination compound 42+ After exhibiting the most promising bactericidal properties, DNA binding (Figure 22) and time-kill kinetics were tested against *E. coli* during exposure to increased concentrations of the complex in a minimal culture medium at 37°C (see Figure 22). Figure 24 An efflux mechanism was observed in the presence of glucose, clearly indicating that this complex can be actively transported in and out of cells. Figure 23 Shown in Figure 26 The Ru content per cell (g) varied at different time intervals. At 5, 10, and 20 minutes, Ru content was generally higher in the presence of glucose, with significant differences observed after 10 minutes.

[0197] Use 4 2+ Treatment of E. coli cells at MIC concentrations for 1, 2, and 24 hours showed that when 4... 2+ Multinucleated cell mitosis occurs at this time. Figure 27 AC and Figure 27 This indicates that in the presence of this complex, cell death is caused by DNA damage. Figure 26 In the middle, DAPI and 4 2+ The direct superposition yielded a Pearson colocation constant >0.9. This constant indicates strong colocation, confirming the 4 2+ Targeting Staphylococcus aureus DNA. To confirm... Figure 28 and 27 The observation results were used to determine, using the Ames mutation test, that compared to natural mutation and ultraviolet irradiation, 4 2+ Mutagenic properties of Escherichia coli Figure 26 ). In 4 2+ Significant DNA mutagenesis was observed at MIC levels and above, and at twice the MIC level, it approached the levels observed under ultraviolet irradiation, indicating that 4 2+ Damage to bacterial cell DNA, such as in Figure 29 The observations made during the formation of filaments. Figure 30 It showed that E. coli was used in 4 2+ The processed TEM images showed that the cell membranes remained intact, further demonstrating that cell death occurred through membrane damage rather than membrane dissolution. Nevertheless, signs of plasmolysis were present in the dead cells (e.g., internal cell leakage observed in III), indicating that 4 2+ It can also cause osmotic damage to cells. Figure 31 The membrane damage assay further confirmed the above DNA damage model. This assay used different concentrations of 4 2+ Extracellular ATP was quantified after treatment, compared to the polymyxin control, using 4 2+ The treated cells retained ATP. This is believed to be because the cells require additional levels of ATP to repair damaged DNA.

[0198] 4 2+ Toxicity screening ( Figure 4 According to the above text regarding 4 4+ The description ( Figure 32 D) Completed. 4 2+ It is non-toxic to wax moths at doses up to 80 mg / kg (maximum clinical daily dose of antibiotics). (Compared to 4) 4+ Similarly, the Mantel-Cox log-rank test showed that there was no statistically significant difference in the survival percentage of larvae between the control group and the compound-treated larvae at any concentration. Figure 33 Further toxicity screening data were provided (in this case, in the case of Acinetobacter baumannii AB184), and these tests clearly showed 4 2+ The presence of [a specific substance] eliminated the colonies, as all treated larvae survived for 120 hours of experimental duration, while those not treated with 4 [a specific substance] were eliminated. 2+ The treated larvae did not survive. A Mantel-Cox log-rank study showed a one-star significant difference in survival percentage between untreated and treated infected larvae. Furthermore, Figure 25 Figures A and 33B show the bacterial CFU counts (Acinetobacter baumannii AB184 bacteria) from extracted larval hemolymph. It is clearly visible from these figures that at both treatment concentrations (40 or 80 mg / kg), the CFU counts were significantly higher than those of other bacteria. 2+ After 48 hours, all colonies were eliminated. This elimination was achieved with a single compound dose. In contrast, in the absence of 4... 2+ Under these conditions, bacterial growth was observed to be exponential.

[0199] Comparison of properties of mononuclear and binuclear complexes

[0200] To further consider compound 1 4+ Up to 4 4+ and compound 1 2+ Up to 4 2+ Antimicrobial activity, selected compound 4 4+ and 4 2+ The results of studies on various microorganisms are shown in Table 8 below. The determination of MIC and MBC values ​​is as described above.

[0201] Table 8. MIC and MBC results of binuclear and mononuclear complexes

[0202]

[0203]

[0204]

[0205] It can be seen that the claimed complexes exhibit activity against a broad library of bacterial strains. In particular, both complexes show high activity against all bacteria, including those identified as a Level 1 priority by the World Health Organization: key carbapenem-resistant strains.

[0206] Tables 9 to 11 further compare coordination compound 4 4+ and 4 2+ Activity compared to clinical standard gentamicin and cisplatin.

[0207] Table 9. Overview of biofilm activities of binuclear and mononuclear complexes against Gram-negative biofilms

[0208]

[0209]

[0210] It can be seen that both compounds are active against the tested Gram-negative biomembranes, indicating that they can penetrate and disrupt biomembranes. (Dinuclear complex 4) 4+ The activity is higher than 4 2+ Or the clinical standard antibiotic gentamicin.

[0211] Table 10. Mutagenesis test results of binuclear and mononuclear complexes

[0212]

[0213] Table 10 shows the mutagenic frequency observed in the untreated control group (natural mutagenesis) at concentrations above the compound's MIC. This mutagenic frequency is lower than that observed with cisplatin. Therefore, these compounds are confirmed to be non-mutagenic for mammalian DNA.

[0214] Table 11. Mammalian cytotoxicity results of binuclear and monouclear complexes

[0215]

[0216] Table 11 shows that the mammalian cytotoxicity data of both compounds are lower than that of cisplatin, with binuclear 4 being the most toxic. 4+ It is more than 10 times less toxic to healthy eukaryotic cells than this well-known drug. Furthermore, 4... 4+ The average treatment index is >60, 4 2+ The average therapeutic index is approximately 6. In comparison, the therapeutic index of cisplatin is 2.

[0217] Complex 4 2+ and 4 4+ The relative uptake rate. For example... Compound As shown, it was observed that relative to complex 4 4+The increased initial uptake rate is attributed to the lower molecular weight of the mononuclear complex.

[0218] Table 12 shows the kinetic solubility of the complexes, for 4... 2+ and 4 4+ The compounds were tested and compared with a soluble positive control (nicardipine). Both compounds passed DMPK analysis and demonstrated optimal solubility and kinetic stability.

[0219] Table 12: Kinetic Turbidity and Solubility of DMPK

[0220] Concentration indicated μM Pass / Fail Buffer Log S Solubility μM Pass 4 2+ ]] 200 pH 7.4 Pass 0.8281 6.7 4 2+ ]] 200 pH 7.4 Pass 0.7961 6.3 4 4+ ]] 200 pH 7.4 Pass 1.138 13.7 4 4+ ]] 200 pH 7.4 Nicardipine 1.138 13.7 Pass 200 pH 7.4 Nicardipine 1.319 20.8 Pass 200 pH 7.4 Nicardipine 1.319 20.8 Pass 200 pH 7.4 ​ 1.319 20.8

Claims

1. Use of a compound in the preparation of an antimicrobial agent, said compound being of formula (I) or formula (Ia): in, X1, X2, X3, and X4 are each N; Y1 and Y2 are each independently selected from: N or C(R) a ); M1 and M2 are each metal centers, with M1 and M2 each being ruthenium; R1, R2 and R a Each is hydrogen; R3 and R4 are each independently selected from: hydrogen, methyl, halogen or a combination thereof; A1, A2, A3, and A4 are each bidentate ligands; and Rings D1 and D2 each independently contain one or more elements selected from N, C(R) a The heteroatom of ); wherein the bidentate ligand is a compound according to formula (III): in, Z1 and Z2 are each N; and R5, R6 and R7 are each independently selected from: hydrogen, methyl, phenyl, methoxy, nitro or a combination thereof.

2. The use according to claim 1, wherein the compound is used as an antibiotic.

3. The use according to claim 2, wherein the compound is used against Gram-negative bacteria.

4. The use according to any one of the preceding claims, wherein Y1 and Y2 are each N.

5. The use according to any one of claims 1 to 3, wherein R5, R6 and R7 are each independently selected from: hydrogen, methyl and phenyl.

6. The use according to any one of claims 1 to 3, wherein A1, A2, A3 and A4 are each independently selected from (1) to (7) or combinations thereof:

7. The use according to claim 1, wherein the compound has a structure according to formula (IV) or formula (IVa): in R8, R8', R8”, R8”’, R9, R9’, R9”, R9”’, R 10 R 10 '、R 10 "、R 10 ”'、R 11 R 11 '、R 11 "、R 11 ”'、R 12 R 12 '、R 12 "、R 12 ”'、R 13 R 13 '、R 13 "and R 13 Each is independently selected from: hydrogen, methyl, methoxy, and phenyl; The conditions are R8, R8', R8”, R8”', R9, R9', R9”, R9”', R 10 R 10 '、R 10 "、R 10 ”'、R 11 R 11 '、R 11 "、R 11 ”'、R 12 R 12 '、R 12 "、R 12 ”'、R 13 R 13 '、R 13 "and R 13 At least one of the following is selected from: methyl, methoxy, and phenyl; and Where R 14 R 15 R 16 R 19 R 20 and R 21 Each is independently selected from: hydrogen, halogens, or combinations thereof.

8. The use according to claim 7, wherein: R8, R8', R8”, and R8”' are the same; R9, R9', R9”, and R9”' are the same; R 10 R 10 '、R 10 "and R 10 Same; R 11 R 11 '、R 11 "and R 11 Same; R 12 R 12 '、R 12 "and R 12 Same; R 13 R 13 '、R 13 "and R 13 "'same.

9. The use according to claim 7 or 8, wherein the compound or a pharmaceutically acceptable salt thereof has a structure according to formula (V) or formula (Va):

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