Antimicrobial conjugated oligoelectrolytes and methods thereof

Abstract

The present disclosure relates generally to antimicrobial conjugated oligoelectrolytes and methods of use thereof. The compounds are useful in the treatment of bacterial infections or bacterial diseases.

Description

Technical Field

[0001] The present disclosure generally relates to antimicrobial conjugated oligoelectrolytes and methods of using the same. Background Art

[0002] Antimicrobial resistance has been recognized as one of the most serious public health problems today. The overuse of antibiotics and the challenges of drug development have exacerbated this challenge. Therefore, there is an urgent need to develop new classes of antibiotics that can combat multidrug-resistant (MDR) priority pathogens that cannot be treated with existing options. Membrane-active molecules have become an attractive platform for the development of such new antibiotics due to their rapid bactericidal activity and low resistance to drug resistance. For example, membrane-soluble antimicrobial peptides (AMPs), as naturally occurring bioactive compounds, have become more popular due to their bactericidal activity and extremely low likelihood of developing drug resistance due to their membrane permeability mode of action. However, AMPs have inherent challenges including proteolytic instability and high synthesis costs. To overcome these challenges, synthetic efforts have been made, including the design of cationic polymers, synthetic peptide oligomers and amphiphilic compounds, all of which have the key membrane-specific mechanisms of AMPs. The core of all these developments is to control the hydrophobic-hydrophilic balance to design molecules that can penetrate and modify bacterial membranes.

[0003] It would be desirable to solve or alleviate at least one of the above problems, or at least provide a useful alternative. Summary of the Invention

[0004] The present disclosure relates to a compound of formula (I) or a pharmaceutically acceptable salt, solvate or isomer thereof:

[0005]

[0006] in

[0007] Each of R1, R2 and R3 is independently selected from H, optionally substituted alkyl, optionally substituted alkoxy;

[0008] Each phenylene ring contains at least one

[0009] wherein each R4 is independently selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, and optionally substituted cycloalkenyl;

[0010] n is an integer selected from 2 to 10; and

[0011] X is a linking group configured to maintain the π-π conjugation between the two phenylene rings.

[0012] In some embodiments, X is selected from:

[0013] in represents a bond to any phenylene ring.

[0014] The present disclosure relates to a compound of formula (IIa) or a pharmaceutically acceptable salt, solvate or isomer thereof:

[0015]

[0016] in

[0017] Each R1 is independently selected from H, optionally substituted alkyl, and optionally substituted alkoxy;

[0018] Each R2 is independently selected from H, optionally substituted alkyl, and optionally substituted alkoxy;

[0019] each R3 is independently selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, and optionally substituted cycloalkenyl; and

[0020] n is an integer selected from 4 to 10.

[0021] In some embodiments, R1 is independently optionally substituted methoxy and R2 is H.

[0022] In some embodiments, at least one R4 is optionally substituted aryl.

[0023] In some embodiments, the compound of Formula (I) or the compound of Formula (IIa) is characterized by the trans configuration or the E configuration.

[0024] In some embodiments, the compound of formula (I) or the compound of formula (IIa) is selected from:

[0025]

[0026] In some embodiments, the compound of Formula (I) or the compound of Formula (IIa) is characterized by a solubility in aqueous media of about 2 μg / mL to 200 μg / mL.

[0027] In some embodiments, the compound of Formula (I) or the compound of Formula (IIa) is characterized by a solubility in aqueous media of about 4 μg / mL to 138 μg / mL.

[0028] In some embodiments, the compound of Formula (I) or the compound of Formula (IIa) is characterized by a minimum inhibitory concentration (MIC) against a microorganism of about 0.5 μg / mL to 64 μg / mL.

[0029] In some embodiments, the compound of Formula (I) or the compound of Formula (IIa) is characterized by a minimum inhibitory concentration (MIC) against Gram-negative bacteria of about 0.5 μg / mL to 64 μg / mL.

[0030] In some embodiments, the compound of Formula (I) or the compound of Formula (IIa) is characterized by a minimum inhibitory concentration (MIC) against Gram-positive bacteria of about 0.5 μg / mL to 16 μg / mL.

[0031] In some embodiments, the MIC is not affected by the presence of proteins and / or salts in the aqueous medium.

[0032] In some embodiments, the compound of formula (I) or the compound of formula (IIa) is characterized by an IC 50 It is about 5 μg / mL to 256 μg / mL.

[0033] In some embodiments, the compound of Formula (I) or the compound of Formula (IIa) is characterized by a selectivity index (IC 50 / MIC) is from about 2 to about 100.

[0034] In some embodiments, the compound of Formula (I) or the compound of Formula (IIa) is characterized by a minimum bactericidal concentration (MBC) of about 0.5 μg / mL to 64 μg / mL.

[0035] In some embodiments, the microorganism does not develop drug resistance after contact with the compound of Formula (I) or the compound of Formula (IIa).

[0036] In some embodiments, bacterial membranes in bacterial cells are depolarized upon contact with a compound of Formula (I) or a compound of Formula (IIa).

[0037] The present disclosure also relates to pharmaceutical compositions comprising a compound of formula (I) or a compound of formula (IIa) or a pharmaceutically acceptable salt, solvate or isomer thereof, optionally in combination with an excipient.

[0038] The present disclosure also relates to compositions comprising a compound of formula (I) or a compound of formula (IIa) or a salt, solvate or isomer thereof, optionally in combination with an excipient.

[0039] The present disclosure also relates to a method for treating bacterial infection or bacterial disease, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I) or a compound of formula (IIa) or a pharmaceutically acceptable salt, solvate or isomer thereof.

[0040] The present disclosure also relates to a compound of formula (I) or a compound of formula (IIa) or a pharmaceutically acceptable salt, solvate or isomer thereof for use in treating bacterial infection or bacterial disease.

[0041] The present disclosure also relates to the use of a compound of formula (I) or a compound of formula (IIa) or a pharmaceutically acceptable salt, solvate or isomer thereof in the preparation of a medicament for treating bacterial infection or bacterial disease.

[0042] In some embodiments, about 0.05 mg / kg to about 10 mg / kg of a compound of Formula (I) or a compound of Formula (IIa), or a pharmaceutically acceptable salt, solvate, or isomer thereof, is provided to a subject in need thereof.

[0043] In some embodiments, the bacteria in the bacterial infection or bacterial disease are antibiotic-resistant bacteria.

[0044] In some embodiments, the bacteria in the bacterial infection or bacterial disease are gram-negative or gram-positive bacteria.

[0045] In some embodiments, the bacteria is selected from Escherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), Klebsiella pneumonia (K. pneumonia), Acinetobacter baumanii (A. baumanii), Staphylococcus aureus (S. aureus), Methicillin-resistant S. aureus (Methicillin-resistant S. aureus), Enterococcus faecalis (E. facium), Enterococcus faecium (E. facium) and non-tuberculous mycobacteria.

[0046] In some embodiments, the bacterial infection or bacterial disease is a bacterial skin infection or a foodborne infection.

[0047] The present disclosure also relates to a method of disinfecting a surface comprising contacting a compound of formula (I) with the surface.

[0048] In some embodiments, the surface is an abiotic surface. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Embodiments of the present invention will now be described by way of non-limiting examples with reference to the accompanying drawings, in which:

[0050] Figure 1 Shown are (a) a schematic diagram of the modular subunits of the antimicrobial COE; (b) the general structure of previously studied quaternary ammonium COEs; and (c) the TPP analogs in this study.

[0051] Figure 2 The synthetic route of phosphonium COE is shown.

[0052] Figure 3The time-kill kinetic curves of DM6P at different concentrations against MRSA (a) and PAO1 (b) are shown. The dashed line indicates the threshold at which 99.9% of bacteria are killed. The detection limit = 100 cfu / mL.

[0053] Figure 4 Serial subcultures of bacteria treated with sub-MIC concentrations of DM6P showed no development of resistance (two independent biological replicates for each compound). Data are plotted relative to the MIC value at the start of the experiment.

[0054] Figure 5 Shown are the changes in membrane permeability of bacterial cells (PAO1 (Gram-negative, left panel) and MRSA (Gram-positive, right panel) over time at different concentrations (4 μg / mL for PAO1 and 1 μg / mL for MRSA) set based on the MIC value. (a) Membrane depolarization assay. The increase in fluorescence intensity correlates with the change in membrane potential. Values are normalized to the complete depolarization induced by 0.2% (v / v) Triton-X-100. (b) Cytoplasmic membrane permeability measured by propidium iodide (30 μM) influx.

[0055] Figure 6 The results of live-dead staining using SYTO-9 / PI (5 μM / 30 μM, respectively) of Pseudomonas aeruginosa treated with DM6P at a concentration of 2×MIC (8 μg / mL) for 30 minutes are shown. Scale bar = 2 μm.

[0056] Figure 7 Shown are representative TEM images of MRSA cells treated with DM6P. Cells were incubated with 4 mg / mL (4×MIC) of the compound for 30 minutes. White arrows indicate regions of membrane destabilization. The white regions are characterized by the absence of electron-dense intracellular regions.

[0057] Figure 8 Figure 2 shows the in vivo efficacy of representative TPP-COE in a mouse wound model. Mice (n=4) were treated with 0.05 mg / kg or 0.25 mg / kg of DM6P to inoculate MRSA. Mice were sacrificed 24 hours later and colonies on agar plates were counted (p<0.0005, one-way ANOVA compared to saline control groups). The dotted line represents the inoculation cfu (6.02) per wound.

[0058] Figure 9The in vivo efficacy of a representative TPP-COE against Staphylococcus pseudointermedius (S. pseudointermedius) was shown, which was treated with 0.1 mg / kg or 0.25 mg / kg of DM6P, compared with an untreated vehicle control group. Bacterial loads were sampled and plated on agar plates 12 hours after treatment (p < 0.05 compared to saline control, one-way ANOVA).

[0059] Figure 10 The in vivo efficacy of a representative TPP-COE against Staphylococcus pseudintermedius is shown, which was treated with 0.1 mg / kg or 0.25 mg / kg of DM6P, compared with an untreated vehicle control group. Mice were sacrificed 24 hours later and colonies on agar plates were counted (p < 0.05 compared to saline control group, one-way ANOVA).

[0060] Figure 11 The structures of other compounds of the present disclosure are shown. DETAILED DESCRIPTION

[0061] The present disclosure is based on the recognition that certain conjugated oligoelectrolytes (COEs) have previously been shown to have antimicrobial activity. They are characterized by a linear hydrophobic π-conjugated core flanked by nitrogen-based cationic side groups that promote binding to negatively charged microbial cell membranes ( Figure 1 a) Molecular topology, hydrophobicity, and the chemical properties of the cationic groups provide structural factors that influence interactions with the lipid bilayer and, in turn, confer bactericidal properties. Existing research has focused on balancing the hydrophobic domains of the pendant subunits and the spacer subunits to enhance bacterial selectivity. Previous work has also demonstrated that increasing molecular length, while reducing overall solubility, can enhance activity by increasing cell binding.

[0062] Cationic groups in antimicrobial COEs are primarily limited to quaternary ammonium (QA) functional groups, which present challenging drug motifs due to potential off-target interactions. This common problem with ammonium functional groups has led to the investigation of other cationic species in drug design. Without wishing to be bound by theory, the inventors propose that benzyltriphenylphosphonium salts may possess antimicrobial activity, and that membrane-targeting motifs functionalized with triphenylphosphonium moieties may serve as antimicrobial agents with superior activity to existing lead compounds.

[0063] Thus, the present disclosure relates to a class of COEs that utilize a cationic triarylphosphonium (TPP) functional group to achieve solubility in aqueous media and compares their antimicrobial efficacy with that of structurally related quaternary ammonium analogs ( Figure 1 b. Figure 1 c and Figure 9 ). The inventors considered that the antimicrobial activity could be improved by enhancing its lipophobicity, a factor that has a key influence on the ability of better compounds to penetrate cell membranes. In addition, the delocalization effect of the charge on the larger TPP part is expected to change the membrane interaction properties, which may affect the selectivity. In order to alleviate the complex problems that may be caused by reduced water solubility, the inventors also studied chemical substitutions on the diphenylethylene core. Interference with the π-π interactions between hydrophobic diphenylethylene units is expected to reduce the tendency of molecules to aggregate and become a viable strategy to improve bioavailability. For example, methoxy substituents can be selected (Scheme 1c, R=OMe series). Alternatively, the alkenylene linking group can be modified. Based on these structural modifications, the inventors studied the effect of TPP cationic substitution on the antimicrobial activity of existing diphenylethylene COE skeletons by systematically studying their activity spectrum against multiple bacterial strains. The effect of the lipophilic group on the interaction of COE with bacterial membranes was also studied. Finally, the in vivo efficacy against community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) and Staphylococcus pseudintermedius was verified in a mouse skin infection model.

[0064] The present disclosure relates to a compound of formula (I) or a pharmaceutically acceptable salt, solvate or isomer thereof:

[0065]

[0066] in

[0067] Each of R1, R2 and R3 is independently selected from H, optionally substituted alkyl, optionally substituted alkoxy;

[0068] Each phenylene ring contains at least one or

[0069] wherein each R4 is independently selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, and optionally substituted cycloalkenyl;

[0070] n is an integer selected from 2 to 10; and

[0071] X is a linking group configured to maintain the π-π conjugation between the two phenylene rings.

[0072] At least one of R1, R2 and R3 on each phenylene ring is Therefore, there can be one, two or three phenylene rings on each ring. Any one of R1, R2 and R3 on each phenylene ring may independently be In some embodiments, at least one of R1, R2, and R3 on each phenylene ring is

[0073] The π(pi) system of the molecule consists of sp 2 The π bond is formed by the interaction of unhybridized p atomic orbitals on sp hybridized and sp hybridized atoms. This interaction, which leads to the formation of π bonds, occurs between p orbitals connected by σ bonds and takes the form of a lateral overlap of the p orbitals.

[0074] In some embodiments, X is selected from:

[0075]

[0076] in represents a bond to any phenylene ring.

[0077] In some embodiments, the compound of formula (I) is a compound of formula (II) or a pharmaceutically acceptable salt, solvate or isomer thereof:

[0078]

[0079] in

[0080] Each of R1, R2 and R3 is independently selected from H, optionally substituted alkyl, optionally substituted alkoxy;

[0081] Each phenylene ring contains at least one

[0082] wherein each R4 is independently selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, and optionally substituted cycloalkenyl;

[0083] n is an integer selected from 2 to 10.

[0084] The present disclosure relates to a compound of formula (IIa) or a pharmaceutically acceptable salt, solvate or isomer thereof:

[0085]

[0086] in

[0087] Each R1 is independently selected from H, optionally substituted alkyl, and optionally substituted alkoxy;

[0088] Each R2 is independently selected from H, optionally substituted alkyl, and optionally substituted alkoxy;

[0089] each R4 is independently selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, and optionally substituted cycloalkenyl; and

[0090] n is an integer selected from 4 to 10.

[0091] In some embodiments, the compound of formula (I) is a compound of formula (III) or a pharmaceutically acceptable salt, solvate or isomer thereof:

[0092]

[0093] in

[0094] Each of R1, R2 and R3 is independently selected from H, optionally substituted alkyl, optionally substituted alkoxy;

[0095] Each phenylene ring contains at least one

[0096] wherein each R4 is independently selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, and optionally substituted cycloalkenyl;

[0097] n is an integer selected from 3 to 10.

[0098] In some embodiments, the compound of formula (III) is a compound of formula (IIIa) or a pharmaceutically acceptable salt, solvate or isomer thereof:

[0099]

[0100] in

[0101] Each R1 is independently selected from H, optionally substituted alkyl, and optionally substituted alkoxy;

[0102] Each R2 is independently selected from H, optionally substituted alkyl, and optionally substituted alkoxy;

[0103] each R4 is independently selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, and optionally substituted cycloalkenyl; and

[0104] n is an integer selected from 2 to 10.

[0105] In some embodiments, the compound of formula (I) is a compound of formula (IV) or a pharmaceutically acceptable salt, solvate, or isomer thereof:

[0106]

[0107] in

[0108] Each of R1, R2 and R3 is independently selected from H, optionally substituted alkyl, optionally substituted alkoxy;

[0109] Each phenylene ring contains at least one

[0110] wherein each R4 is independently selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, and optionally substituted cycloalkenyl;

[0111] n is an integer selected from 2 to 10.

[0112] In some embodiments, the compound of formula (IV) is a compound of formula (IVa) or a pharmaceutically acceptable salt, solvate, or isomer thereof:

[0113]

[0114] in

[0115] Each R3 is independently selected from H, optionally substituted alkyl, and optionally substituted alkoxy;

[0116] each R4 is independently selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, and optionally substituted cycloalkenyl; and

[0117] n is an integer selected from 3 to 10.

[0118] "Alkyl" refers to a monovalent alkyl group which may be linear or branched, preferably having 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms. Examples of such alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, n-hexyl, and the like.

[0119] "Alkoxy" refers to the group alkyl-O-, wherein the alkyl group is as described above. Examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.

[0120] "Aryl" refers to an unsaturated aromatic carbocyclic group having a single ring (e.g., phenyl) or multiple condensed rings (e.g., naphthyl or anthracenyl), preferably having 6 to 14 carbon atoms. Examples of aryl include phenyl, naphthyl, and the like.

[0121] "Heteroaryl" refers to a monovalent aromatic heterocyclic group that satisfies Hückel's aromaticity rules (i.e., contains 4n+2 π electrons), preferably having 2 to 10 carbon atoms and 1 to 4 heteroatoms selected from oxygen, nitrogen, selenium, and sulfur (including oxides of sulfur, selenium, and nitrogen) within the ring. Such heteroaryl groups may have a single ring (e.g., pyridyl, pyrrolyl, or its N-oxide, or furyl) or multiple fused rings (e.g., indolizinyl, benzimidazolyl, coumarinyl, quinolinyl, isoquinolinyl, or benzothiophenyl).

[0122] Examples of heteroaryl groups include, but are not limited to, oxazole, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, isothiazole, phenoxazine, phenothiazine, thiazole, thiadiazole, oxadiazole, oxatriazole, tetrazole, thiophene, benzo[b]thiophene, triazole, imidazopyridine, and the like.

[0123] "Cycloalkyl" refers to a cyclic alkyl group having a single ring or multiple fused rings, preferably containing 3 to 11 carbon atoms. Such cycloalkyl groups include, but are not limited to, monocyclic structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclooctyl, etc., or polycyclic structures such as adamantyl, indanyl, 1,2,3,4-tetrahydronaphthyl, etc.

[0124] "Heterocyclyl" refers to a monovalent saturated or unsaturated group having a single ring or multiple fused rings, preferably containing 1 to 8 carbon atoms and 1 to 4 heteroatoms selected from nitrogen, sulfur, oxygen, selenium or phosphorus in the ring. The most preferred heteroatom is nitrogen. It should be understood that when, for example, R2 or R' is an optionally substituted heterocyclyl having one or more than one ring heteroatom, the heterocyclyl can be connected to the core molecule of the compound of the present invention through a C-C bond or a C-heteroatom bond, particularly a C-N bond.

[0125] Examples of heterocyclic and heteroaryl groups include, but are not limited to, oxazole, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, isothiazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, indoline, phthalimide, 1,2,3,4-tetrahydroisoquinoline, 4,5,6,7-tetrahydrobenzo[b]thiophene, thiazole, thiadiazole, oxadiazole, oxatriazole, tetrazole, thiazolidine, thiophene, benzo[b]thiophene, morpholine, piperidinyl, pyrrolidine, tetrahydrofuranyl, triazole, and the like.

[0126] In the present specification, "optionally substituted" means that the group may or may not be substituted or fused (thereby forming a fused polycyclic group) with one or more than one group selected from the group consisting of hydroxy, acyl, alkyl, alkoxy, alkenyl, alkenyloxy, alkynyl, alkynyloxy, amino, aminoacyl, thio, aralkyl, aralkyloxy, aryl, aryloxy, carboxyl, amido, cyano, halogen, nitro, phosphono, sulfo, phosphorylamino, phosphinyl, heteroaryl, heteroaralkyl, heteroaryloxy, heterocyclyl, hetero Cycloalkyl, heterocyclyloxy, oxyacyl, oxime, oxime ether, hydrazone, oxyacylamino, oxysulfonylamino, aminoacyloxy, trihalomethyl, trialkylsilyl, pentafluoroethyl, trifluoromethoxy, difluoromethoxy, trifluoromethylthio, trifluorovinyl, mono- and di-alkylamino, mono- (substituted alkyl) amino and di- (substituted alkyl) amino, mono- and di-arylamino, mono- and di-heteroarylamino, mono- and di-heteroarylamino, and unsymmetrical disubstituted amines having different substituents selected from alkyl, aryl, heteroaryl, and heterocyclyl groups, etc., and may also include a bond to a solid support material (e.g., substituted onto a polymer resin). For example, an "optionally substituted amino" group may include amino acids and peptide residues.

[0127] The compounds described herein may contain one or more asymmetric centers and may therefore exist in a variety of isomeric forms, such as enantiomers and / or diastereomers. For example, the compounds described herein may exist in the form of single enantiomers, diastereomers, or geometric isomers, or in the form of mixtures of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. One skilled in the art can separate isomers from a mixture by known methods, including chiral high performance liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or prepare preferred isomers by asymmetric synthesis. The present invention also encompasses compounds described herein as single isomers that are substantially free of other isomers, and alternatively, compounds described herein as mixtures of various isomers. As used herein, "optically enriched" means that the compound is composed of a significantly higher proportion of one enantiomer. In certain embodiments, the compounds of the present invention are composed of at least about 90% by weight of a preferred enantiomer. In other embodiments, the compound is composed of at least about 95%, 98% or 99% by weight of the preferred enantiomer. The preferred enantiomer can be separated from the racemic mixture by any method known to those skilled in the art, including chiral high performance liquid chromatography (HPLC) and the formation and crystallization of chiral salts, or prepared by asymmetric synthesis. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33: 2725 (1977); Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, Tables of Resolving Agents and Optical Resolutions p. 268 (ELEliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972).

[0128] Conjugated oligoelectrolytes (COEs) are a class of molecules defined by a hydrophobic conjugated core with terminal polar ionic side groups. In certain embodiments, the hydrophobic and hydrophilic portions of a COE can be rationally designed into the molecule to correspond to the organization of hydrophilic and hydrophobic domains in a lipid bilayer. This structural design typically involves only an unbranched internal structure with charged groups at both ends, which facilitates the spontaneous insertion of the COE into the cell membrane through electrostatic and hydrophobic interactions between the COE and the lipids.

[0129] The compounds of the present invention have a substantially linear topology. Topology refers to the molecular structure of a compound within the constraints of three-dimensional (3D) space. This linear topology has two nodes at the ends and no connecting nodes. This linear topology facilitates lipid membrane embedding.

[0130] In some embodiments, each R1 is independently selected from H and optionally substituted alkoxy. In some embodiments, each R1 is independently selected from H, optionally substituted methoxy, optionally substituted ethoxy, and optionally substituted propoxy. In some embodiments, each R1 is independently selected from H, optionally substituted methoxy, and optionally substituted C4-C 10 An alkoxy group having a phosphonium moiety at its terminal end, wherein the phosphonium moiety may be a triarylphosphonium moiety.

[0131] In some embodiments, each R2 is independently selected from H and optionally substituted alkoxy. In some embodiments, each R2 is independently selected from H, optionally substituted methoxy, optionally substituted ethoxy, and optionally substituted propoxy. In some embodiments, each R2 is independently selected from H, optionally substituted methoxy, and optionally substituted C4-C 10 An alkoxy group having a phosphonium moiety at its end, wherein the phosphonium moiety may be a triarylphosphonium moiety.

[0132] In some embodiments, each R3 is independently selected from H and optionally substituted alkoxy. In some embodiments, each R3 is independently selected from H, optionally substituted methoxy, optionally substituted ethoxy, and optionally substituted propoxy. In some embodiments, each R3 is independently selected from H, optionally substituted methoxy, and optionally substituted C4-C 10 An alkoxy group having a phosphonium moiety at its end, wherein the phosphonium moiety may be a triarylphosphonium moiety.

[0133] In some embodiments, R1 is independently optionally substituted methoxy and R2 is H.

[0134] As mentioned above, the compounds of the present disclosure contain at least one terminal polar ionic side group at each end. The ionic side group can be an optionally substituted C4-C ... 10 The ionic side groups may be located at R1, R2 and / or R3.

[0135] In some embodiments, at least one of R1, R2, and R3 on each phenylene ring is or In some embodiments, at least two of R1, R2, and R3 on each phenylene ring are or In some embodiments, R3 on each phenylene ring is or In some embodiments, R3 on a phenylene ring is or And R1 and R2 on other phenylene rings are or In some embodiments, R1 and R2 on each phenylene ring are or

[0136] In some embodiments, at least one of R1, R2, and R3 on each phenylene ring is In some embodiments, at least two of R1, R2, and R3 on each phenylene ring are In some embodiments, R3 on each phenylene ring is In some embodiments, R3 on a phenylene ring is And R1 and R2 on other phenylene rings are In some embodiments, R1 and R2 on each phenylene ring are

[0137] In some embodiments, each R4 is independently selected from optionally substituted aryl and optionally substituted heteroaryl. In some embodiments, each R3 is optionally substituted aryl. In some embodiments, each R3 is optionally substituted phenyl.

[0138] In some embodiments, at least one R4 is optionally substituted aryl. In some embodiments, at least two R4 are optionally substituted aryl. In this case, the two aryl moieties can be connected to a single phosphonium ion, or each aryl moiety can be connected to each phosphonium ion. In some embodiments, at least three R4 are optionally substituted aryl. In some embodiments, at least four R4 are optionally substituted aryl. In some embodiments, at least five R3 are optionally substituted aryl.

[0139] In some embodiments, n is an integer selected from 2 to 9 or 3 to 9. In some embodiments, n is an integer selected from 4 to 9. In some embodiments, n is an integer selected from 4 to 8. In some embodiments, n is an integer selected from 4, 6, and 8.

[0140] In some embodiments, the compounds of formula (I), (II), (III), and (IV) are characterized by the trans or E configuration.

[0141] In some embodiments, the compound of formula (I), the compound of formula (II), the compound of formula (III), and the compound of formula (IV) are selected from:

[0142]

[0143] In some embodiments, the compound of formula (II) is selected from:

[0144]

[0145] In some embodiments, the compound of formula (I), formula (II), formula (III), and formula (IV) are characterized by a solubility in aqueous media of about 2 μg / mL to about 200 μg / mL. In some embodiments, the solubility is about 2 μg / mL to about 180 μg / mL, about 2 μg / mL to about 160 μg / mL, about 2 μg / mL to about 140 μg / mL, about 2 μg / mL to about 120 μg / mL, or about 2 μg / mL to about 100 μg / mL. In some embodiments, the compound of formula (I) is characterized by a solubility in aqueous media of about 4 μg / mL to about 138 μg / mL.

[0146] As used herein, the term "aqueous medium" refers to a water-based solvent or solvent system that primarily comprises water. Such solvents can be polar or non-polar, and / or protic or aprotic. A solvent system refers to the combination of solvents that results in a final single phase. "Solvent" and "solvent system" may include, but are not limited to, pentane, cyclopentane, hexane, cyclohexane, benzene, toluene, dioxane, chloroform, ether, dichloromethane, tetrahydrofuran, ethyl acetate, acetone, dimethylformamide, acetonitrile, dimethyl sulfoxide, nitromethane, propylene carbonate, formic acid, butanol, isopropyl alcohol, propanol, ethanol, methanol, acetic acid, ethylene glycol, diethylene glycol, or water. A water-based solvent or solvent system may also contain dissolved ions, salts, and molecules such as amino acids, proteins, carbohydrates, and phospholipids. Such salts include, but are not limited to, sodium chloride, potassium chloride, ammonium acetate, magnesium acetate, magnesium chloride, magnesium sulfate, potassium acetate, potassium chloride, sodium acetate, sodium citrate, zinc chloride, HEPES sodium salt, calcium chloride, ferric nitrate, sodium bicarbonate, potassium phosphate, and sodium phosphate. Thus, biological fluids, physiological solutions, and culture media also fall within this definition.

[0147] In some embodiments, the compounds of formula (I), formula (II), formula (III), and formula (IV) are characterized in that the minimum inhibitory concentration (MIC) for microorganisms is from about 0.5 μg / mL to about 64 μg / mL. In other embodiments, the minimum inhibitory concentration (MIC) is from about 0.5 μg / mL to about 60 μg / mL, from about 0.5 μg / mL to about 55 μg / mL, from about 0.5 μg / mL to about 50 μg / mL, from about 0.5 μg / mL to about 45 μg / mL, from about 0.5 μg / mL to about 40 μg / mL, from about 0.5 μg / mL to about 35 μg / mL, or from about 0.5 μg / mL to about 30 μg / mL.

[0148] In some embodiments, the compound of formula (I), formula (II), formula (III), and formula (IV) are characterized in that the minimum inhibitory concentration (MIC) for Gram-negative bacteria is about 0.5 μg / mL to 64 μg / mL. In other embodiments, the minimum inhibitory concentration (MIC) is about 0.5 μg / mL to about 60 μg / mL, about 0.5 μg / mL to about 55 μg / mL, about 0.5 μg / mL to about 50 μg / mL, about 0.5 μg / mL to about 45 μg / mL, about 0.5 μg / mL to about 40 μg / mL, about 0.5 μg / mL to about 35 μg / mL, or about 0.5 μg / mL to about 30 μg / mL.

[0149] In some embodiments, the compounds of formula (I), (II), (III), and (IV) are characterized in that the minimum inhibitory concentration (MIC) against Gram-positive bacteria is from about 0.5 μg / mL to about 16 μg / mL. In other embodiments, the minimum inhibitory concentration (MIC) is from about 0.5 μg / mL to about 15 μg / mL, from about 0.5 μg / mL to about 14 μg / mL, from about 0.5 μg / mL to about 13 μg / mL, from about 0.5 μg / mL to about 12 μg / mL, from about 0.5 μg / mL to about 11 μg / mL, from about 0.5 μg / mL to about 10 μg / mL, or from about 0.5 μg / mL to about 9 μg / mL.

[0150] In some embodiments, the presence of protein and / or salt in the aqueous medium does not affect the MIC value.

[0151] In some embodiments, the compounds of formula (I), formula (II), formula (III), and formula (IV) are characterized by an IC of 100 for mammalian cells. 50 In other embodiments, the IC 50From about 5 μg / mL to about 300 μg / mL, about 5 μg / mL to about 280 μg / mL, about 5 μg / mL to about 260 μg / mL, about 5 μg / mL to about 250 μg / mL, about 5 μg / mL to about 240 μg / mL, about 5 μg / mL to about 220 μg / mL, about 5 μg / mL to about 200 μg / mL, about 5 μg / mL to about 180 μg / mL, about 5 μg / mL to about 160 μg / mL, about 5 μg / mL to about 140 μg / mL, about 5 μg / mL to about 120 μg / mL, about 5 μg / mL to about 100 μg / mL, about 5 μg / mL to about 80 μg / mL, about 5 μg / mL to about 60 μg / mL, about 5 μg / mL to about 40 μg / mL, or about 5 μg / mL to about 20 μg / mL.

[0152] In some embodiments, the compounds of formula (I), (II), (III), and (IV) are characterized by a selectivity index (IC 50 / MIC) is from about 2 to about 100. In other embodiments, the selectivity index is from about 2 to about 90, from about 2 to about 80, from about 2 to about 70, from about 2 to about 60, from about 2 to about 50, from about 2 to about 40, from about 2 to about 30, from about 2 to about 20, or from about 2 to about 15.

[0153] In some embodiments, the compounds of formula (I), (II), (III), and (IV) are characterized by a minimum bactericidal concentration (MBC) of about 0.5 μg / mL to about 64 μg / mL. In other embodiments, the minimum inhibitory concentration (MIC) is about 0.5 μg / mL to about 60 μg / mL, about 0.5 μg / mL to about 55 μg / mL, about 0.5 μg / mL to about 50 μg / mL, about 0.5 μg / mL to about 45 μg / mL, about 0.5 μg / mL to about 40 μg / mL, about 0.5 μg / mL to about 35 μg / mL, or about 0.5 μg / mL to about 30 μg / mL.

[0154] In some embodiments, after exposure to a compound of formula (I), (II), (III) and / or (IV), the microorganism does not develop drug resistance. The drug resistance may be undetectable.

[0155] In some embodiments, the bacterial membrane of the bacterial cell is depolarized upon contact with a compound of Formula (I), a compound of Formula (II), a compound of Formula (III), and / or a compound of Formula (IV).

[0156] The present disclosure also relates to pharmaceutical compositions comprising a compound of formula (I), a compound of formula (II), a compound of formula (III) and / or a compound of formula (IV) or a pharmaceutically acceptable salt, solvate or isomer thereof, optionally in combination with a pharmaceutically acceptable excipient.

[0157] The present disclosure also relates to compositions comprising a compound of formula (I), a compound of formula (II), a compound of formula (III) and / or a compound of formula (IV), or a salt, solvate or isomer thereof, optionally in combination with an excipient.

[0158] The present disclosure also relates to a method for treating a bacterial infection or bacterial disease, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I), a compound of formula (II), a compound of formula (III) and / or a compound of formula (IV) or a pharmaceutically acceptable salt, solvate or isomer thereof.

[0159] The present disclosure also relates to a compound of formula (I), a compound of formula (II), a compound of formula (III) and / or a compound of formula (IV) or a pharmaceutically acceptable salt, solvate or isomer thereof for use in treating bacterial infection or bacterial disease.

[0160] The present disclosure also relates to the use of a compound of formula (I), a compound of formula (II), a compound of formula (III) and / or a compound of formula (IV) or a pharmaceutically acceptable salt, solvate or isomer thereof in the preparation of a medicament for treating bacterial infection or bacterial disease.

[0161] In some embodiments, about 0.05 mg / kg to about 10 mg / kg of a compound of Formula (I), (II), (III), and / or (IV), or a pharmaceutically acceptable salt, solvate, or isomer thereof, is provided to a subject in need thereof. In other embodiments, the concentration is from about 0.05 mg / kg to about 9.5 mg / kg, about 0.05 mg / kg to about 9 mg / kg, about 0.05 mg / kg to about 8.5 mg / kg, about 0.05 mg / kg to about 8 mg / kg, about 0.05 mg / kg to about 7.5 mg / kg, about 0.05 mg / kg to about 7 mg / kg, about 0.05 mg / kg to about 6.5 mg / kg, about 0.05 mg / kg to about 6 mg / kg, about 0.05 mg / kg to about 5.5 mg / kg, about 0.05 mg / kg to about 5 mg / kg, about 0.05 mg / kg to about 4.5 mg / kg, about 0.05 mg / kg to about 4 mg / kg, about 0.05 mg / kg to about 3.5 mg / kg, or about 0.05 mg / kg to about 3 mg / kg.

[0162] In some embodiments, the bacteria in the bacterial infection or bacterial disease are antibiotic-resistant bacteria.

[0163] In some embodiments, the bacteria in the bacterial infection or bacterial disease are gram-negative or gram-positive bacteria.

[0164] In some embodiments, the bacteria is selected from Escherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), Klebsiella pneumonia (K. pneumonia), Acinetobacter baumanii (A. baumanii), Staphylococcus aureus (S. aureus), Methicillin-resistant S. aureus (Methicillin-resistant S. aureus), Enterococcus faecalis (E. facium), and non-tuberculous mycobacteria. In some embodiments, the bacterium is selected from the group consisting of Acetobacter aurantius, Acinetobacter baumannii, Actinomyces israelii, Agrobacterium radiobacter, Agrobacterium tumefaciens, Anaplasma, Anaplasma phagocytophilum, Azorhizobium caulinodans, Azotobacter vinelandii, viridans streptococci, Bacillus, Bacillus anthracis, Bacillus brevis, Bacillus cereus, Bacillus fusiformis, Bacillus licheniformis, and Bacillus genus. licheniformis), Bacillus megaterium, Bacillus mycoides, Bacillus stearothermophilus, Bacillus subtilis, Bacillus thuringiensis, Bacteroides, Bacteroides fragilis, Bacteroides gingivalis, Bacteroides melaninogenicus (now called Prevotella melaninogenicus),melaninogenica), Bartonella, Bartonella henselae, Bartonella quintana, Bordetella, Bordetella bronchiseptica, Bordetella pertussis, Borrelia burgdorferi, Brucella, Brucella abortus, Brucella melitensis, Brucella suis, Burkholderia, Burkholderia mallei, Burkholderia pseudomallei, Burkholderia cepacia, Calymmatobacterium granulomatis), Campylobacter, Campylobacter coli, Campylobacter fetus, Campylobacter jejuni, Campylobacter pylori, Chlamydia, Chlamydia trachomatis, Chlamydophila, Chlamydophila pneumoniae (formerly Chlamydia pneumoniae), Chlamydophila psittaci (formerly Chlamydia psittaci), Clostridium, Clostridium botulinum, Clostridium difficile, Clostridium perfringens (formerly Clostridium welchii), Clostridium tetanitetani), Corynebacterium, Corynebacterium diphtheria, Corynebacterium fusiforme, Coxiella burnetii, Ehrlichia chaffeensis, Ehrlichia ewingii, Eikenella corrodens, Enterobacter cloacae, Enterococcus, Enterococcus avium, Enterococcus durans, Enterococcus faecalis, Enterococcus faecium, Enterococcus gallinarum, Enterococcus maloratus, Escherichia coli coli), Fusobacterium necrophorum, Fusobacterium nucleatum, Gardnerella vaginalis, Haemophilus, Haemophilus ducreyi, Haemophilus influenzae, Haemophilus parainfluenzae, Haemophilus pertussis, Haemophilus vaginalis, Helicobacter pylori, Klebsiella pneumonia, Lactobacillus, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus casei, Lactococcus lactis, Legionella pneumophila pneumophila), Leptospira interrogans, Leptospira noguchinoguchii), Listeria monocytogenes, Methanobacterium extroquens, Microbacterium multiforme, Micrococcus luteus, Moraxella catarrhalis, Mycobacterium, Mycobacterium avium, Mycobacterium bovis, Mycobacterium diphtheria, Mycobacterium intracellulare, Mycobacterium leprae, Mycobacterium lepraemurium, Mycobacterium phlei, Mycobacterium smegmatis, Mycobacterium tuberculosis tuberculosis), Mycoplasma, Mycoplasmafermentans, Mycoplasma genitalium, Mycoplasma hominis, Mycoplasma penetrans, Mycoplasma pneumonia, Mycoplasma Mexicana, Neisseria, Neisseria gonorrhoeae, Neisseria meningitides, Pasteurella, Pasteurella multocida, Pasteurella tularensis, Peptostreptococcus, Porphyromonas gingivalis, Prevotella melaninifera melaninogenica) (formerly known as Bacteroides melaninogenicus), Pseudomonas aeruginosaaeruginosa), Rhizobium radiobacter, Rickettsia, Rickettsia prowazekii, Rickettsia psittaci, Rickettsia Quintana, Rickettsia rickettsia, Rickettsia trachomae, Rochalimaea, Rochalimaea henselae, Rochalimaea Quintana, Rothia dentocariosa, Salmonella, Salmonella enteritidis, Salmonella typhi, Salmonella typhimurium typhimurium), Serratia marcescens, Shigella dysenteriae, Spirillum volutans, Staphylococcus, Staphylococcus aureus, Staphylococcus epidermidis, Stenotrophomonas maltophilia, Streptococcus, Streptococcus agalactiae, Streptococcus avium, Streptococcus bovis, Streptococcus cricetus, Streptococcus faceium, Streptococcus faecalis, Streptococcus ferus, Streptococcus gallinarum gallinarum), Streptococcus lactis, Streptococcus mitis, Streptococcus mutansmutans), Streptococcus oralis, Streptococcus pneumonia, Streptococcus pyogenes, Streptococcus rattus, Streptococcus salivarius, Streptococcus sanguis, Streptococcus sobrinus, Treponema, Ureaplasma urealyticum, Vibrio, Vibrio cholera, Vibrio comma, Vibrio parahaemolyticus, Vibrio vulnificus vulnificus), Wolbachia, Yersinia, Yersinia enterocolitica, Yersinia pestis, and Yersinia pseudotuberculosis.

[0165] In some embodiments, the bacterial infection or bacterial disease is a bacterial skin infection. The bacterial skin infection may include, but is not limited to, cellulitis, erysipelas, folliculitis, impetigo, carbuncle, or furuncle. In some embodiments, the bacterial infection or bacterial disease is a foodborne infection. The foodborne infection is an inflammation of the stomach and intestines that may cause diarrhea, nausea, vomiting, abdominal pain, abdominal cramps, chills, and / or fever. In some embodiments, the bacterial infection or bacterial disease is selected from brucellosis, Campylobacter infection, cat scratch disease, cholera, Escherichia coli infection, gonorrhea, Klebsiella infection, Enterobacter infection and Serratia infection, Legionella infection, meningococcal infection, pertussis, plague, Pseudomonas infection, Salmonella infection, shigellosis, typhoid fever, tularemia, anthrax, Clostridium difficile colitis, diphtheria, enterococcal infection, erysipelothrixosis, listeriosis, nocardiosis, pneumococcal infection, Staphylococcal infection, Streptococcal infection, non-venereal syphilis (Bejel), yaws and pinta, leptospirosis, Lyme disease, rat-bite fever, relapsing fever, syphilis, tuberculosis, actinomycosis, Bacteroides infection, botulism, Clostridium infection and tetanus.

[0166] In some embodiments, the bacterial infection or bacterial disease is characterized by antibiotic resistance. The antibiotic resistance may be antibiotic resistance to antibiotic drugs such as vancomycin, daptomycin, ceftaroline, linezolid, ceftazidime, aminoglycosides, carbapenems, and / or streptogramins.

[0167] In some embodiments, the compound of formula (I), the compound of formula (II), the compound of formula (III), and / or the compound of formula (IV), or a pharmaceutically acceptable salt, solvate, or isomer thereof, is administered in combination with an antibiotic. The administration of the compound of formula (I), or a pharmaceutically acceptable salt, solvate, or isomer thereof, and the antibiotic may be performed simultaneously or sequentially.

[0168] The present disclosure also relates to a method of disinfecting a surface comprising contacting a compound of formula (I), a compound of formula (II), a compound of formula (III), and / or a compound of formula (IV) with the surface.

[0169] In some embodiments, the surface is an abiotic surface.

[0170] The compounds of the present invention can be administered to a subject in the form of their pharmaceutically acceptable salts. Suitable pharmaceutically acceptable salts include, but are not limited to, salts of pharmaceutically acceptable inorganic acids, such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, boric acid, sulfamic acid, and hydrobromic acid; or salts of pharmaceutically acceptable organic acids, such as acetic acid, propionic acid, butyric acid, tartaric acid, maleic acid, hydroxymaleic acid, fumaric acid, maleic acid, citric acid, lactic acid, mucic acid, gluconic acid, benzoic acid, succinic acid, oxalic acid, phenylacetic acid, methanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, salicylic acid, sulfanilic acid, aspartic acid, glutamic acid, edetic acid, stearic acid, palmitic acid, oleic acid, lauric acid, pantothenic acid, tannic acid, ascorbic acid, and valeric acid.

[0171] Basic salts include, but are not limited to, salts formed with pharmaceutically acceptable cations, such as salts of sodium, potassium, lithium, calcium, magnesium, ammonium, and alkylammonium. In particular, the present invention includes within its scope cationic salts of phosphate groups, such as sodium or potassium salts, or alkyl esters (e.g., methyl, ethyl) of phosphate groups.

[0172] It should be understood that any compound that is a prodrug of a compound of formula (I), a compound of formula (II), a compound of formula (III), and / or a compound of formula (IV) also falls within the scope and spirit of the present invention. Therefore, the compounds of the present invention can be administered to a subject in the form of a pharmaceutically acceptable prodrug. The term "prodrug" is used in its broadest sense to encompass derivatives that are converted into the compounds of the present invention in vivo. Such derivatives will be apparent to those skilled in the art. Other texts generally describing prodrugs (and their preparation) include: Design of Prodrugs, 1985, H. Bundgaard (Elsevier); The Practice of Medicinal Chemistry, 1996, Camille G. Wermuth et al., Chapter 31 (Academic Press); and A Textbook of Drug Design and Development, 1991, Bundgaard et al., Chapter 5 (Harwood Academic Publishers).

[0173] The compounds of the present invention may exist in crystalline form, either as free compounds or as solvates (eg hydrates), and both forms are intended to fall within the scope of the present invention. Methods of solvation are generally known in the art.

[0174] The compound of the present invention or its pharmaceutically acceptable salt, solvate or prodrug is administered to the patient in a therapeutically effective amount. As used herein, a therapeutically effective amount is intended to include at least partially achieving the desired effect, or delaying the onset of the bacterial disease, or inhibiting its progression, or preventing or reversing its progression.

[0175] As used herein, the term "effective amount" relates to the amount of the compound providing the desired therapeutic activity when used according to a desired dosage regimen. Administration can be carried out at intervals of minutes, hours, days, weeks, months or years, or continuously in any of these time periods. Suitable dosage can be in the range of about 0.1 nanograms per kilogram of body weight to 1 gram per kilogram of body weight per dosage, for example, in the range of 1 milligram to 1 gram per kilogram of body weight per dosage. In one embodiment, dosage can be in the range of 1 milligram to 500 milligrams per kilogram of body weight per dosage. In another embodiment, dosage can be in the range of 1 milligram to 250 milligrams per kilogram of body weight per dosage. In yet another embodiment, dosage can be in the range of 1 milligram to 100 milligrams per kilogram of body weight per dosage, for example, up to 50 milligrams per kilogram of body weight per dosage.

[0176] The appropriate dosage and administration regimen can be determined by the attending physician and may depend on the severity of the condition and the general age, health, and weight of the patient to be treated. Other factors include the activity of the specific compound employed, age, weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the specific disease being treated. The amount of the compound in the disclosed compositions also depends on the specific compound in the composition.

[0177] The compounds of the present invention can be administered in single or multiple doses. While the active ingredient can be administered alone, it is preferably provided as a composition, more preferably a pharmaceutical composition. The formulation of such compositions is well known to those skilled in the art. The compositions may contain any suitable carrier, diluent, or excipient. This includes all conventional solvents, dispersion media, fillers, solid carriers, coatings, antifungal and antibacterial agents, transdermal absorption enhancers, surfactants, isotonic agents, and absorbents. It should be understood that the compositions of the present invention may also contain other supplementary physiologically active agents.

[0178] The carrier is pharmaceutically "acceptable", i.e., compatible with the other ingredients of the composition and not harmful to the patient. The composition can be conveniently provided in unit dosage form and can be prepared by any method known in the art of pharmacy. Such methods include the step of mixing the active ingredient with the carrier which constitutes one or more excipients. Generally, the composition is prepared by uniformly and intimately mixing the active ingredient with a liquid carrier or a finely divided solid carrier, or both, and then shaping the product if necessary.

[0179] The composition defined above can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, orally, vaginally, or by means of an implanted reservoir. As used herein, the term "parenteral" includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. Preferably, the composition is administered orally, intraperitoneally, or intravenously. The sterile injectable formulations of the compositions disclosed herein can be aqueous suspensions or oily suspensions, which can be prepared using suitable dispersants or wetting agents and suspending agents according to techniques known in the art. The sterile injectable formulation can also be a sterile injectable solution or suspension in a non-toxic, parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Acceptable carriers and solvents that can be used include water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils are conventionally used as solvents or suspending media.

[0180] For this purpose, any bland fixed oil may be used, including synthetic mono- or diglycerides. Fatty acids such as oleic acid and its glyceride derivatives may be used to prepare injections, as may natural pharmaceutically acceptable oils such as olive oil or castor oil, especially in their polyoxyethylated forms. These oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants, such as carboxymethyl cellulose or similar dispersants commonly used in the preparation of pharmaceutically acceptable dosage forms (including emulsions and suspensions). Other commonly used surfactants (such as Tweens, Spans and other emulsifiers) or bioavailability enhancers commonly used in the preparation of pharmaceutically acceptable solid, liquid or other dosage forms may also be used for formulation purposes.

[0181] The pharmaceutically acceptable compositions defined above can be orally administered in any orally acceptable dosage form, including but not limited to capsules, tablets, aqueous suspensions, or aqueous solutions. For tablets for oral administration, common carriers include lactose and corn starch. Lubricants such as magnesium stearate are also typically added. When administered orally in capsule form, useful diluents include lactose and dry corn starch. When an aqueous suspension for oral administration is desired, the active ingredient is combined with an emulsifier and a suspending agent. If desired, certain sweeteners, flavorings, or coloring agents may also be added.

[0182] The pharmaceutical composition for parenteral injection can include pharmaceutically acceptable sterile aqueous or non-aqueous solution, dispersion, suspension or emulsion, and can be reconstituted into the sterile powder of sterile injectable solution or dispersion before use. Suitable aqueous and non-aqueous carriers, diluents, solvents or supporting agents include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters (such as ethyl oleate). Suitable fluidity can be maintained, for example, by using coating materials (such as lecithin), by maintaining the required particle size in the case of dispersions, and by using surfactants.

[0183] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifiers, and dispersants. Protection against the action of microorganisms can be ensured by the addition of various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, and the like. The addition of isotonic agents, such as sugars, sodium chloride, and the like, may also be desirable. Prolonged absorption of injectable pharmaceutical dosage forms can be achieved by the addition of agents that delay absorption, such as aluminum monostearate and gelatin.

[0184] If desired, and for more effective distribution, the compounds can be incorporated into slow-release or targeted-delivery systems such as polymer matrices, liposomes, and microspheres.

[0185] The injectable formulations can be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved or dispersed in sterile water or other sterile injectable medium immediately before use.

[0186] Alternatively, the pharmaceutically acceptable compositions as defined above may be in the form of suppositories for rectal administration. These may be prepared by mixing the agent with a suitable non-irritating excipient which is solid at room temperature but liquid at rectal temperature and which therefore melts in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycol.

[0187] The pharmaceutically acceptable compositions as defined above may also be administered topically, particularly when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, skin or lower intestinal tract. Suitable topical formulations may be readily prepared for such areas or organs.

[0188] Topical application for the lower intestinal tract can be achieved in a rectal suppository formulation (see above) or in a suitable enema formulation.Topical transdermal patches may also be used.

[0189] For topical application, the pharmaceutically acceptable composition can be formulated into a suitable ointment containing an active ingredient suspended or dissolved in one or more than one carrier. Carriers for topical application of the compound as defined above include, but are not limited to, mineral oil, liquid paraffin, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsifying wax, and water. Alternatively, the pharmaceutically acceptable composition can be formulated into a suitable lotion or cream containing an active ingredient suspended or dissolved in one or more than one pharmaceutically acceptable carrier. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, and water.

[0190] For ophthalmic use, the pharmaceutically acceptable composition can be formulated as a micronized suspension in isotonic and pH-adjusted sterile saline, or more preferably as a solution in isotonic and pH-adjusted sterile saline with or without a preservative such as benzalkonium chloride. Alternatively, for ophthalmic use, the pharmaceutically acceptable composition can be formulated in an ointment such as petrolatum.

[0191] The pharmaceutically acceptable compositions defined above may also be administered by nasal aerosol or inhalation. Such compositions may be prepared according to techniques known in the art of pharmaceutical formulation and may be formulated as solutions in saline, using benzyl alcohol or other suitable preservatives, absorption enhancers to improve bioavailability, fluorocarbons, and / or other conventional solubilizing or dispersing agents.

[0192] Most preferably, the pharmaceutically acceptable compositions as defined above can be formulated for oral administration. Such formulations can be administered with or without food. In some embodiments, the pharmaceutically acceptable compositions as defined above can be administered without food. In other embodiments, the pharmaceutically acceptable compositions as defined above can be administered with food.

[0193] It should be understood that, in addition to the active ingredients specifically mentioned above, the compositions or combinations of the present invention may include conventional other agents for the types of compositions or combinations, such as those suitable for oral administration, which may include such other agents as binders, sweeteners, thickeners, flavorings, disintegrants, coatings, preservatives, lubricants and / or delay agents. Suitable sweeteners include sucrose, lactose, glucose, aspartame or saccharin. Suitable disintegrants include corn starch, methylcellulose, polyvinyl pyrrolidone, xanthan gum, bentonite, alginic acid or agar. Suitable flavorings include peppermint oil, wintergreen oil, cherry flavor, orange flavor or raspberry flavor. Suitable coating agents include homopolymers or copolymers of acrylic acid and / or methacrylic acid and esters thereof, waxes, fatty alcohols, zein, shellac or gluten. Suitable preservatives include sodium benzoate, vitamin E, alpha-tocopherol, ascorbic acid, methylparaben, propylparaben or sodium bisulfite. Suitable lubricants include magnesium stearate, stearic acid, sodium oleate, sodium chloride or talc. Suitable delay agents include glyceryl monostearate or glyceryl distearate.

[0194] Example

[0195] Synthesis and characterization of compounds

[0196] The synthesis of TPP-containing COEs is based on the reaction of the corresponding neutral alkyl iodide precursors with triphenylphosphine (which replaces the tertiary amine), see Figure 2 . This method was verified by synthesizing D4P, D6P and D8P from the reported 1a, 1b and 1c, respectively. Using commercially available isoeugenol as the starting material, the key functionalized stilbene core 2 was constructed through olefin metathesis reaction with excellent catalyst loading efficiency (0.05 mol%), reflecting the advantages of green synthesis strategy in the construction of new drug skeletons. Compound 2 was alkylated with the corresponding α,ω-alkyl diiodide to obtain compounds 3a to 3c, and the neutral intermediate was further reacted with triphenylphosphine to obtain the target TPP compound. Good yields were obtained after sequential recrystallization from toluene and dichloromethane. The specific synthesis and purification methods are detailed below. The ease of synthetic diversification and relatively simple purification make this type of compound attractive for further derivatization.

[0197] DM4P was obtained from N,N-dimethylformamide and dichloromethane by fractional recrystallization to obtain high-quality single crystals suitable for X-ray diffraction to determine its chemical structure. Crystal analysis showed that the distance between phosphorus atoms is The total length of the molecule is These molecular size parameters work synergistically with the hydrophobic force to facilitate membrane embedding.

[0198] The sufficient water solubility of antibiotics is a key drug-like property to achieve good bioavailability, accurate drug concentration and desired pharmacological effect. The solubility of all TPP derivatives was determined in phosphate buffered saline (PBS) (Table S3). Solubility generally decreases with increasing molecular length, and the characteristic solubility of the stilbene derivatives D4P, D6P and D8P is 76 μg / mL, 18 μg / mL and 4 μg / mL, respectively, and has a value estimated by turbidimetric screening (turbidimetric screening) less than 10 μg / mL). Consistent with our design strategy, the functionalization modification of the stilbene core improves overall solubility. The solubility of the best soluble derivative DM4P in PBS is 138 μg / mL, while the values observed for DM6P and DM8P are 108 μg / mL and 4 μg / mL, respectively. This difference can be attributed to the destruction of the face-to-face stacking effect of the molecule and the increase in the total polar surface area, while keeping it below the estimated value required for spontaneous insertion into the membrane.

[0199] Activity against a broad spectrum of pathogens

[0200] Antimicrobial activity screening was performed against a panel of clinically relevant ESKAPE pathogens using minimum inhibitory concentration (MIC) determinations in cation-adjusted Mueller-Hinton broth (CA-MHB). The results of these studies are presented in Table 1. In contrast to D4 (>64 μg / mL for both S. aureus and E. coli), the TPP-modified analog D4P exhibited better antimicrobial activity (4 μg / mL for S. aureus and E. coli). The increased lipophilicity of the TPP group likely compensated for the previously identified weak interaction of D4 with bacteria. D6P exhibited slightly better activity against Gram-positive SA strains (2 μg / mL) compared to D6P's TMA counterpart D6 (32 μg / mL). DM6P achieved a further reduction of the MIC to 1 μg / mL for all Gram-positive pathogens. Overall, these results indicate that TPP analogs exhibit comparable or superior activity against Gram-positive bacteria compared to their QA counterparts.

[0201] Table 1. MIC values of lead COEs against various ESKAPE pathogens. The highest concentration tested was 128 μg / mL for D4, polymyxin B, and vancomycin, and 64 μg / mL for the remaining compounds. Due to turbidity issues caused by solubility, the maximum concentration tested for D8P and DM8P was 4 μg / mL. EC = Escherichia coli K12 (ATCC 10798), PA = Pseudomonas aeruginosa (ATCC 47085), KP (CRE) = Klebsiella pneumoniae (carbapenem-resistant Enterobacteriales) (ATCC BAA1705), AB = Acinetobacter baumannii (ATCC 17978), SA = Staphylococcus aureus (ATCC 29213), MRSA = methicillin-resistant Staphylococcus aureus (ATCC BAA1717), FAE = Enterococcus faecalis OG1RF (ATCC 47077), FAC = Enterococcus faecium (ATCC 19434).

[0202]

[0203] Note: For both DM8P and D8P, the molecules remain fully soluble at 4 μg / ml. At higher concentrations, accurate comparison becomes difficult due to precipitation.

[0204] The difference in activity conferred by the TPP group was even more pronounced against Gram-negative bacteria. For D6P, an MIC of 8 μg / ml was observed against P. aeruginosa, while the QA analog D6 had an MIC of >64 μg / ml against this bacterium. The antimicrobial activity of DM6P was further enhanced, with MIC values reaching 4 to 8 μg / ml against the key WHO-defined Gram-negative pathogens Acinetobacter baumannii, Klebsiella pneumoniae, and P. aeruginosa. In summary, these two chemical modifications together resulted in a greater than 16-fold increase in antimicrobial activity. In contrast, D8 had only limited effect against E. coli, which can be attributed to the higher permeability of the E. coli membrane compared to other difficult-to-treat Gram-negative bacteria such as Pseudomonas species and Acinetobacter species.

[0205] In general, changing the terminal group from trimethylalkylammonium to TPP improves overall antimicrobial activity, likely due to an overall increase in cationic lipophilicity while retaining the dicationic nature of the molecule, which is essential for solubility and enhanced binding. However, as shown in Table 1, and in contrast to previous observations from studies with D4, D6, and D8, examination of the D4P to D8P analogs suggests that increasing compound length is not necessarily a direct approach to reducing the minimum MIC. The lower dependence on molecular length compared to previously studied structures suggests that other potential membrane disruption mechanisms may be relevant to bioactivity.

[0206] We also investigated nontuberculous mycobacteria (NTM). These opportunistic pathogens are of increasing clinical importance due to their increasing prevalence and difficulty in treatment. The mycobacterial membrane is an inherently stronger barrier to antibiotic penetration due to its higher lipid content relative to other pathogens and the high lipophilicity imparted by long-chain mycolic acids. We also evaluated the antimicrobial activity of TPP analogs against mycobacteria in Middlebrook 7H9 broth. The characteristic MIC of DM4P and DM6P against the clinical target of Mycobacterium abscessus (M. abscessus) was 8 μg / ml, which is comparable to values observed with currently used antibiotics (i.e., amikacin and linezolid) (Table 2). Therefore, activity against this group of bacteria suggests a potential avenue for developing COEs as antibiotics against challenging drug targets.

[0207] Table 2. MICs of TPP COE against Mycobacterium abscessus (ATCC 19977)

[0208]

[0209] To investigate the extent to which the antimicrobial activity of COE-phosphonium derivatives can be modulated by the chemical properties of the phosphonium fragment, we synthesized more hydrophilic phosphonium variants, viz. Figure 2 In a DM6THPP variant, the more hydrophobic phenyl substituent was replaced with a hydroxypropyl group to enhance the compound's solubility. As expected, the compound had extremely high solubility in water (>2560 μg / mL). However, DM6THPP had significantly reduced activity against the tested pathogenic bacterial strains (MIC>64 μg / mL, see Table 1), highlighting that the nature of the phosphonium functional group provides a relevant regulatory structural factor that modulates the antimicrobial effect.

[0210] TPP substitution does not adversely increase cytotoxicity

[0211] The cytotoxicity of TPP COE against mammalian cell line (A549) was also tested. After replacing the TMA group with TPP, the IC 50 The change in the values was negligible (Table 3). Compared with D8, the selectivity index (IC 50 / MIC) was higher, reaching 11, while D8 had similar antibacterial activity, but its IC 50 Based on these results, we chose to focus on DM6P in most of our subsequent studies to investigate the interactions of TPP-COE with biological systems.

[0212] Table 3. IC values for mammalian cell line A549 (lung epithelial cells) 50Selectivity index is based on IC 50 Determined by dividing by the MIC value against MRSA.

[0213]

[0214] Antimicrobial activity in complex environments

[0215] The complexity of the physiological environment often influences drug activity, particularly when cations in solution contribute to oligomerization-dependent mechanisms of action, as is the case with daptomycin. Binding of the free drug to plasma proteins effectively reduces the concentration of free drug available for bacterial killing, leading to the observed reduction in activity. We tested the MIC values of DM4P and DM6P in the presence of human serum albumin (40 g / L) and high salt concentrations (Table 4). Under these conditions, antimicrobial activity was minimally affected by these environmental factors, with the MIC remaining within a 2-fold range.

[0216] Table 4. MICs of major COEs in a simulated physiological environment. HSA = human serum albumin.

[0217]

[0218] TPP-COE has bactericidal activity

[0219] Determination of biological activity by MIC measurement cannot distinguish between inhibition of bacterial replication and direct bacterial killing. To determine the extent to which TPP-COE effectively kills bacteria, bacterial cells treated with varying concentrations of DM6P were plated on tryptone soy agar (TSA) plates, and the minimum bactericidal concentration (MBC) that completely inactivated the inoculum was determined (Table 5). MBC values ranged from 2 μg / mL for MRSA to 8 μg / mL for PAO1 and remained within a range of 2 to 4 times the MIC for all strains tested.

[0220] Table 5. Minimum bactericidal concentrations (MBCs) of DM6P against different pathogens. EC = Escherichia coli K12 (ATCC 10798), PA = Pseudomonas aeruginosa (ATCC 47085), KP (CRE) = Klebsiella pneumoniae (ATCC BAA1705), AB = Acinetobacter baumannii (ATCC 17978), MRSA = Methicillin-resistant Staphylococcus aureus (ATCC BAA1717), FAE = Enterococcus faecalis OG1RF (ATCC 47077).

[0221]

[0222] We treated MRSA and Pseudomonas aeruginosa cultures with different concentrations of DM6P and took aliquots at specific intervals to quantify the number of remaining bacteria. Figure 3 As shown in Figure 2, when the concentration was higher than the MBC value of 4 μg / mL, MRSA was almost completely killed (<100 cfu / mL) within the first two hours. For Pseudomonas aeruginosa, after processing for 24 hours at the MIC, the bacterial load was reduced by 99.95% (3.5 log). Reaching the MBC value or exceeding the treatment under the MBC value resulted in the complete elimination of cells. In short, these results show that TPP derivatives retain the unique bactericidal activity of COEs.

[0223] TPP-COE minimally induced resistance development

[0224] Unlike traditional antibiotics, the nonspecific effects of membrane-active antibiotics minimize the development of their own resistance because of the huge metabolic burden involved. Based on this premise, we subcultured bacteria with DM6P or ciprofloxacin (control) at 0.5 times the MIC to study the adaptation of bacteria to external antibiotic pressure. In the control experiment, MRSA produced resistant mutants within 4 generations after treatment with ciprofloxacin, see Figure 4 a. Figure 4 b shows a slower rate of development of P. aeruginosa. In contrast, DM6P induced no observable resistance mutants against either bacterial strain under these experimental conditions. Combined with its broad-spectrum activity, this feature offers the opportunity to use COE antimicrobials to address mixed polymicrobial infections with a lower incidence of antibiotic resistance.

[0225] Mechanism of DM6P's membrane interaction

[0226] The interaction of DM6P with different bacterial cell membranes was first investigated by spectroscopic methods. In normal cells, the compartmentalization of the intracellular space by the lipid bilayer maintains the transmembrane potential required for normal cell division and energy production. The membrane potential-sensitive dye 3,3'-dipropylthiodicarbonylcyanine iodide (DiSC3(5)) accumulates within the membranes of healthy, polarized cells, where its fluorescence is self-quenched. Depolarization of the membrane, which is required to maintain this gradient, causes the release of the dye, resulting in an increase in fluorescence intensity. Treatment of MRSA and PAO1 cells coated with DiSC3(5) with DM6P showed a dose-dependent depolarization of the bacterial membrane upon binding of the molecule ( Figure 5 a), indicating that COE induces ion leakage. Even at half the MIC level, the compound's interaction with the cell membrane appears to induce ion efflux.

[0227] Another sign of cytoplasmic membrane damage is increased permeability to small molecules. Propidium iodide (PI) is a membrane-impermeable organic dye that exhibits a significant increase in fluorescence intensity when bound to intracellular DNA and can therefore be used to study the potential mechanisms of action of antibiotics. Molecular treatments that cause membrane disturbances can induce rapid influx of the dye, resulting in an increase in fluorescence emission over monitoring time. When MRSA and PAO1 were treated with DM6P at concentrations of 0.5 to 2 times the MIC (0.5 μg / mL to 2 μg / mL for MRSA and 2 μg / mL to 8 μg / mL for PA), an increase in emission intensity from the propidium cation was observed ( Figure 5 b). For treated PAO1 cells, the increase in PI influx appears to be concentration-independent. For treated MRSA cells, PI influx was observed at 2× the MIC, corresponding to the bactericidal concentration of DM6P. This discrepancy suggests two plausible concentration-dependent mechanisms of action. Similar observations were made with tobramycin, where the pattern of protein inhibition at low concentrations (<4 μg / mL) was replaced by outer membrane disruption at higher concentrations (8 μg / mL).

[0228] To quantify PI uptake at the cellular level, flow cytometry measurements were performed. PAO1 suspensions were treated with 16 μg / mL (4× MIC) of DM6P and then stained with PI for 15 minutes, a duration at which the majority of the dye was observed to be taken up by the cells. Treatment with a bactericidal concentration of COE demonstrated near-quantitative cellular staining with PI compared to buffer-treated controls. Heat-killed control populations exhibited similar fluorescence intensity compared to COE-treated samples, consistent with increased membrane disorder due to DM6P.

[0229] Visualization of COE-induced membrane damage

[0230] Membrane damage induced by COE treatment was also confirmed by fluorescence microscopy. SYTO-9 in combination with PI is often used as a live-dead stain to distinguish cells with permeabilized membranes. SYTO-9 accumulates in both healthy and dead cells and shows strong fluorescence upon binding to nucleic acids. In combination with PI, the contrast between the colors is used as a visual indicator of membrane damage. PAO1 cells were exposed to 16 μg / mL DM6P for 30 minutes, then stained with both dyes and imaged immediately. Consistent with previous experiments, Figure 6 In the red fluorescence channel, only the COE-treated sample was observed to have obvious PI staining, reflecting the destroyed endometrium.

[0231] The damage of bacterial cell membrane was observed by transmission electron microscopy (TEM). Staphylococcus aureus cells were incubated with 4 μg / mL DM6P for 30 minutes and then immediately fixed to preserve the membrane ultrastructure close to the time of death as much as possible. Figure 7a and Figure 7 As shown in b, the untreated MRSA control showed smooth, well-aligned cell walls with no signs of morphological damage. DM6P treatment resulted in visible cell shrinkage and membrane disintegration, as shown in Figure 7 The area highlighted by the white arrow in c is shown. Portions of the cell wall and cytoplasmic membrane, which can be clearly observed as thick grey lines, were found to be missing. Figure 7 In Figure d, plasma membrane separation was also observed in the cells, a key hallmark of plasmolysis and presumably an early stage of cell death, preceding leakage of intracellular components. These observations are consistent with micrographs of cells treated with the membrane-active AMPs gramicidin S and PGLa. Together, these morphological features are consistent with the central concept that TPP-COE induces cell membrane damage, similar to previously studied COEs. Cellular leakage, disrupted protein localization and inactivation, and small molecule permeation are downstream observations frequently associated with cell membrane perturbations.

[0232] In vivo studies

[0233] To explore the potential of TPP-COE as a viable therapy, we evaluated the efficacy of DM6P in treating bacterial infections in vivo. Due to the prevalent clinical problems associated with Staphylococcus aureus skin and soft tissue infections (SSTIs), a Staphylococcus aureus mouse wound infection model was used. Figure 8 As shown, mice inoculated with MRSA treated with DM6P at doses of 0.05 mg / kg and 0.25 mg / kg showed a 3.8-log and 4.2-log reduction in bacterial load compared to the saline-treated group (over 99.99% at 0.25 mg / kg). The low concentrations required to achieve this activity in vivo and the absence of bacterial regrowth after 24 hours, relative to the current clinical dose of daptomycin of 4 mg / kg to 6 mg / kg, highlight the potential for further development of TPP-COEs with improved activity and safety.

[0234] The physiological compatibility of DM6P was assessed by measuring blood biomarker levels in mouse blood samples after 14 days of treatment at the highest concentration of 0.25 mg / kg. We assessed common biomarkers such as total protein content (TP), albumin (ALB), globulin (GLO), total bilirubin (TBIL), alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), blood urea nitrogen (BUN), and creatinine (CRE) and found no significant differences in protein levels or enzyme activities compared to saline-treated controls (Table 6). Hematoxylin-eosin (H&E) staining of major organs in DM6P-treated mice also did not show any signs of abnormalities and did not impair wound healing.

[0235] Table 6. Blood biomarker levels in mice treated with 0.9% saline (control group) or DM6P (0.25 mg / kg). Data are expressed as mean ± sd (n = 3). Standard ranges are derived from the veterinary database (MNChip, Tianjin). TP = total protein, ALB = albumin, GLO = globulin, TBIL = total bilirubin, ALT = alanine aminotransferase, AST = aspartate aminotransferase, GGT = gamma-glutamyltransferase, BUN = blood urea nitrogen, CRE = creatinine.

[0236]

[0237] Staphylococcus pseudintermedius (SP) is particularly pathogenic in dogs, and skin infections such as pyoderma are characterized by prolonged treatment and high rates of treatment failure due to antimicrobial resistance. Here, we demonstrated in a mouse wound infection model that Figure 11 The compounds described herein have high in vivo antimicrobial efficacy, characterized by a greater than 3-log reduction in bacterial load at low concentrations. Figure 9 and Figure 10 The colony counts at 12 and 24 hours are shown. The group tested with saline had 5.16×10 8 The group tested with DM6P (0.1 mg / kg) had an average CFU of 1.62×10 7 The group tested with DM6P (0.25 mg / kg) had an average CFU of 5.51×10 5 The average CFU / g was 4.37 mmol / L. In addition, the mice did not show any negative side effects after compound administration, indicating tolerability.

[0238] Table 7: Compound 1 to Compound 5 ( Figure 11 ) against methicillin-resistant Staphylococcus aureus (USA300) and Escherichia coli K12 (ATCC 10798)

[0239]

[0240] in conclusion

[0241] A series of COEs bearing a lipophilic TPP group were synthesized to understand how this cationic functional group affects antibiotic activity relative to the more frequently studied quaternary ammonium counterparts. The results showed that replacing the cationic center from TMA to TPP resulted in broad-spectrum activity against ESKAPE plates as well as mycobacteria and remained unaffected in complex physiologically simulated environments. By carefully maintaining the hydrophobic-hydrophilic balance throughout the molecule, DM6P achieved optimal activity against bacteria. Furthermore, this TPP substitution did not compromise cytotoxicity and retained the membrane-perturbing properties of previously studied COEs. Importantly, based on a murine wound model, DM6P was shown to be effective in treating MRSA skin infections, even at a dose of 0.05 mg / kg. This in vivo efficacy was further validated in infections with Staphylococcus pseudintermedius. These findings further demonstrate the straightforward molecular design principles for expanding the spectrum of activity of antibacterial stilbene-derived COEs.

[0242] Synthesis scheme and method

[0243] (E)-4,4'-(ethylene-1,2-diyl)bis(2-methoxyphenol)(2)

[0244] Compound 1 was synthesized according to a modified method from the literature. 2 In a 250 mL round-bottom flask equipped with a magnetic stir bar, 2-methoxy-4-propenylphenol (0.5 g, 1.0 equivalent) and Grubbs II catalyst (1 mg) were added in a glove box. The flask was sealed with a septum and evacuated while maintaining a positive nitrogen flow. The reaction mixture was then heated to 90°C and stirred for 10 minutes, during which time the mixture rapidly solidified. The crude product was suspended with dichloromethane (2 mL) and methanol (3 mL) and stirred for 30 minutes. The resulting suspension was allowed to stand at 0°C for another 30 minutes, filtered, and washed with cold methanol until the filtrate was colorless. The resulting solid was dried under vacuum to obtain pure compound 2.

[0245] 1H NMR (400MHz, DMSO-d6) δ9.02 (s, 2H), 7.13 (d, J=2.0Hz, 2H), 6.96-6.90 (m, 4H), 6.74 (d, J=8.1Hz, 2H), 3.82 (s, 6H).

[0246] General procedure for the alkylation of compound 2.

[0247] Under an inert atmosphere, compound 2 (0.5 g, 1.0 equiv), potassium carbonate (3 equiv), and the corresponding α,ω-diiodoalkane (6 equiv) were dissolved in acetone in a 250 mL round-bottom flask equipped with a magnetic stir bar. The reaction mixture was heated to reflux for 48 hours. After cooling to room temperature, the reaction solution was concentrated under reduced pressure, diluted with dichloromethane, and washed with water (200 mL x 3) and brine (200 mL). The organic portion was dried over sodium sulfate, filtered, and the solvent removed in vacuo. The crude product was purified by column chromatography (silica, elution with n-hexane / dichloromethane 1:2) to obtain the desired pure product.

[0248] (E)-1,2-bis(4-(4-iodobutoxy)-3-methoxyphenyl)ethylene(3a)

[0249] Using 1,4-diiodobutane, the product was obtained as a white solid (73% yield).

[0250] 1H NMR (400 MHz, chloroform-d) δ 7.09-7.02 (m, 2H), 6.94 (s, 1H), 6.87 (d, J = 8.3 Hz, 1H), 4.08 (t, J = 6.2 Hz, 2H), 3.95 (s, 3H), 3.31 (t, J = 6.8 Hz, 2H), 2.13-2.04 (m, 2H), 1.99 (d, J = 8.3 Hz, 2H). 13C NMR (101 MHz, CDCl3) δ 149.66, 148.04, 131.03, 126.74, 119.54, 113.31, 109.27, 67.94, 56.01, 30.22, 30.14, 6.45.

[0251] (E)-1,2-bis(4-((6-iodohexyl)oxy)-3-methoxyphenyl)ethylene (3b)

[0252] Using 1,6-diiodohexane, the product was obtained as a white solid (70% yield).

[0253] 1H NMR (400MHz, chloroform-d) δ7.08 (d, J=2.0Hz, 1H), 7.06-7.01 (m, 1H), 6.94 (s, 1H), 6.88 (d, J=8.3Hz, 1 H), 4.06 (t, J=6.7Hz, 2H), 3.95 (s, 3H), 3.23 (t, J=7.0Hz, 2H), 1.89 (s, 4H), 1.55-1.47 (m, 4H).

[0254] (E)-1,2-bis(4-((8-iodooctyl)oxy)-3-methoxyphenyl)ethylene (3c)

[0255] Using 1,8-diiodooctane, the product was obtained as a white solid (66% yield).

[0256] 1H NMR (400 MHz, chloroform-d) δ 7.05 (d, J = 2.0 Hz, 1H), 7.01 (dd, J = 8.3, 2.0 Hz, 1H), 6.91 (s, 1H), 6.85 (d, J = 8.3 Hz, 1H), 4.03 (t, J = 6.8 Hz, 2H), 3.19 (t, J = 7.0 Hz, 2H), 1.89-1.78 (m, 4H), 1.49-1.30 (m, 8H).

[0257] General procedure for the quaternization of COEs (A)

[0258] Compound 1 (1.0 equiv) and PPh3 (2.5 equiv) were added to a screw cap vial and purged with N2. The reagent was dissolved in 2 mL of DMF and heated to 90°C. After stirring for 48 hours, the reaction mixture was cooled to room temperature and toluene (about 10 mL) was added to cover the liquid surface. The flask was placed in a refrigerator overnight and the white solid was collected by filtration and washed thoroughly with ether. The crude solid was redissolved in DMF and triturated with ether to obtain the pure compound.

[0259] General procedure for the quaternization of COE (B)

[0260] Compound 1 (1.0 equivalents) and trihydroxypropylphosphine (3 equivalents) are loaded into a screw cap vial and purged with N2. The reagent is dissolved in 2ml DMF and heated to 90°C. After stirring for 48 hours, the reaction mixture is cooled to room temperature and 40ml ether is added to precipitate the product as an oily residue. The product is washed with toluene and ether and redissolved in a minimum amount of DMF. The solution is ground twice with ether and dried under high vacuum to obtain the product as a colloidal white solid (50% yield).

[0261] General procedure for the quaternization of COEs (C)

[0262] Compound 1 is loaded into a screw cap vial and dissolved in 1 mL of DMF at 45°C. A solution of trimethylamine in THF (0.5 mL, 3.2 M) is added once and the reaction is stirred for 48 hours. The solvent is removed under reduced pressure and the resulting residue is dissolved in a minimum of methanol. Pure compound is obtained by grinding with ether.

[0263] DM4P

[0264] The reaction was carried out by general procedure A. The product was obtained in 92% yield as a white solid.

[0265] 1H NMR (400MHz, chloroform-d) δ7.78 (s, 9H), 7.70-7.62 (m, 6H), 7.03-6.97 (m, 2H), 6.90 (d, J=1.6Hz, 1H), 6.85 (d, J=8. 3Hz, 1H), 4.12 (t, J=5.5Hz, 2H), 3.83 (d, J=3.4Hz, 2H), 3.69 (s, 3H), 2.21 (s, 2H), 1.93 (d, J=7.9Hz, 2H).13C NMR (101 MHz, chloroform-d) δ 134.92, 134.89, 133.60, 133.50, 130.30, 130.18, 126.30, 119.40, 118.89, 118.04, 113.08, 109.21, 67.04, 55.51, 39.50, 18.80.31P NMR (162 MHz, CDCl3) δ 24.34ESI-MS (TOF): [(M-2I) 2+ ]453.6

[0266] DM6P

[0267] The reaction was carried out by general procedure A. The product was obtained in 86% yield as a white solid.

[0268] 1H NMR (400MHz, DMSO-d6) δ7.95-7.74 (m, 15H), 7.18 (d, J=2.0Hz, 1H), 7.07-7.00 (m, 2H), 6.90 (d, J=8.4 Hz, 1H), 3.92 (t, J=6.4Hz, 2H), 3.79 (s, 3H), 3.58 (s, 2H), 1.67 (s, 2H), 1.55 (s, 4H), 1.44 (s, 2H).13C NMR (101MHz, DMSO-d6) δ134.89, 134.86, 133.62, 133.52, 130.29, 130.16, 126.24, 119.3 9, 118.97, 118.12, 112.97, 109.15, 67.96, 55.48, 39.50, 28.40, 24.70, 21.72, 19.91.31P NMR(162MHz, DMSO-d6)δ24.04.ESI-MS(TOF): [(M-2I) 2+ ]481.6

[0269] DM8P

[0270] The reaction was carried out by general procedure A. The product was obtained in 82% yield as a white solid.

[0271] 1H NMR (400MHz, DMSO-d6) δ7.78 (d, J=1.1Hz, 15H), 7.19 (d, J=1.9Hz, 1H), 7.03 (d, J=6.7Hz, 2H), 6.91 (d, J =8.4Hz, 1H), 3.92 (t, J = 6.5Hz, 2H), 3.80 (s, 3H), 3.56 (s, 2H), 1.68 (s, 2H), 1.38 (d, J = 70.8Hz, 10H).13C NMR (126MHz, DMSO-d6) δ134.48, 134.45, 133.20, 133.12, 129.88, 129.78, 126.14, 119.16, 118. 63, 117.95, 113.68, 110.05, 68.36, 55.62, 28.41, 27.96, 27.57, 24.97, 21.41, 20.53, 20.13.31P NMR(162MHz, DMSO-d6)δ24.08.ESI-MS(TOF): [(M-2I) 2+ ]509.7

[0272] D4P

[0273] The reaction was carried out by general procedure A to afford the product as a white solid in 75% yield.

[0274] 1H NMR (400MHz, chloroform-d) δ7.71 (d, J=1.6Hz, 9H), 7.64-7.57 (m, 6H), 7.32-7.27 (m, 2H), 6.80 (s, 1H), 6.73 (d, J =8.8Hz, 2H), 4.03 (t, J = 5.6Hz, 2H), 3.78 (d, J = 3.5Hz, 2H), 2.19-2.11 (m, 2H), 1.81 (d, J = 7.8Hz, 2H).31P NMR(162MHz, DMSO-d6)δ24.08.13C NMR (101MHz, DMSO-d6) δ158.27, 135.43, 135.40, 134.13, 134.03, 130.80, 130.68, 127.87, 126.26 , 119.39, 118.54, 115.18, 66.47, 29.75, 29.58, 20.45, 19.95, 18.98, 18.95.ESI-MS (TOF): [(M-2I) 2+ ]423.6

[0275] D6P

[0276] The reaction was carried out by general procedure A. The product was obtained as a white solid in 77% yield.

[0277] 1H NMR (400MHz, chloroform-d) δ7.88-7.79 (m, 10H), 7.73 (s, 6H), 7.38 (d, J=8.8Hz, 2H), 6.89 (s, 1H), 6.85 (d, J=8.8Hz, 2H), 3.96 (t, J=6.3Hz, 2H), 3.74 (d, J=3.0Hz, 2H), 1.75 (d, J=2.4Hz, 6H), 1.51 (s, 2H).13C NMR (101MHz, DMSO-d6) δ158.47, 135.39, 135.36, 134.13, 134.03, 130.79, 130.67, 130.39, 127.88, 126. 20, 119.47, 118.62, 115.08, 67.75, 36.27, 30.14, 29.97, 28.82, 25.23, 22.24, 22.20, 20.91, 20.41.31P NMR(162MHz, DMSO-d6)δ24.05.ESI-MS(TOF): [(M-2I) 2+ ]451.6

[0278] D8P

[0279] The reaction was carried out by general procedure A. The product was obtained as a white solid in 81% yield.

[0280] 1H NMR (400MHz, chloroform-d) δ7.82 (s, 9H), 7.73 (d, J=2.8Hz, 6H), 7.40 (d, J=8.9Hz, 2H), 6.91 (s, 1H), 6 .87(d, J=8.8Hz, 2H), 3.95(t, J=6.5Hz, 2H), 3.72(s, 2H), 1.68(s, 5H), 1.53-1.22(m, 8H).13C NMR (126MHz, DMSO-d6) δ134.47, 133.21, 133.13, 129.89, 129.79, 129.74, 127.02, 125.58, 117.9 6, 114.47, 67.34, 39.50, 28.27, 27.97, 27.57, 24.96, 21.42, 20.53, 20.13.ESI-MS(TOF): [(M-2I) 2+ ]479.7

[0281] DM6THPP

[0282] The reaction was carried out by general procedure B. The product was obtained in 50% yield as a white solid.

[0283] 1H NMR (400MHz, DMSO-d6) δ7.20 (d, J=1.9Hz, 2H), 7.10-7.01 (m, 4H), 6.94 (d, J=85Hz, 2H), 4.80 (t, J=5.0Hz, 6H), 3 .97 (t, J=6.4Hz, 4H), 3.82 (s, 6H), 3.48 (q, J=5.7Hz, 12H), 2.25 (d, J=3.3Hz, 16H), 1.66 (s, 16H), 1.47 (s, 12H).

[0284] Single crystal growth and measurement

[0285] X-ray quality single crystals were grown by slow diffusion of hexane into a solution of DM4P dissolved in dichloromethane. Single crystal X-ray diffraction data were collected at 100 K using a Bruker D8Advance diffractometer. Frame integration was performed using the Bruker SAINT software package with the narrow frame algorithm. The molecular formula unit C was analyzed using the Bruker SHELXTL software package using space group P-1. 60 H 60 The structure of O4P2I2·CHCl2, Z = I, was solved and refined. The final anisotropic full-matrix least-squares refinement with 363 variables converged to R1 = 3.65% for the observed data and R2 = 9.92% for all data based on the F2 value.

[0286] Minimum inhibitory concentration determination

[0287] The minimum inhibitory concentration (MIC) of COE was determined by broth microdilution according to the Clinical and Laboratory Standards Institute (CLSI) M07-A10 guidelines. Single colonies were isolated from overnight cultures and grown to mid-logarithmic phase (OD 600 Compounds were prepared as stock solutions in DMSO and diluted in CA-MHB in sterile flat-bottom 96-well plates. Subsequent concentrations were prepared by diluting the initial solution two-fold. An equal volume of bacteria was added to the compound to a final concentration of 5 × 10 5 The plate was incubated at 37°C for 16 to 18 hours and the plate was counted using a microplate reader (TECAN ) in OD 600 MIC 90 The value is determined by the concentration at which less than 10% bacterial growth is observed compared to the corresponding control. The MIC value of each compound is determined in triplicate. For each test, polymyxin B (Gram-negative) or vancomycin (Gram-positive) is used as a positive control.

[0288] Minimum bactericidal concentration (MBC) determination

[0289] Bacterial samples from broth microdilution in 96-well plates were treated with varying concentrations of DM6P according to the method for determining MICs. Colonies were plated onto 20 μL TSB agar plates, which were incubated overnight at 37°C. The MBC was determined as the COE concentration at which no colony growth was observed. All measurements were performed in triplicate.

[0290] Time-kill kinetics

[0291] Time-kill kinetics were determined for Pseudomonas aeruginosa (PAO1, ATCC 47085) and methicillin-resistant Staphylococcus aureus (ATCC BAA-1717). The tested bacteria were transferred to cation-adjusted Mueller Hinton II broth (CAMHB, BD Biosciences), cultured overnight at 37°C and 200 rpm shaking to mid-logarithmic phase, and diluted to OD 600 A value of 0.2 was set and aliquoted into 2 mL aliquots in culture tubes. The resulting bacterial suspensions were then treated with varying concentrations of DM6P and incubated at 37°C with shaking at 200 rpm. At regular intervals, a 100 μL aliquot of each sample was diluted 10-fold in PBS, and 100 μL of this inoculum was then plated onto TSB agar plates. Colonies were incubated overnight at 37°C and counted to determine the cfu / mL of each sample. The experiment was repeated three times.

[0292] Membrane depolarization assay

[0293] The degree of bacterial cell membrane depolarization was determined using the voltage-sensitive dye DiSC3 (5) (ThermoFisher, USA). The dye accumulates in polarized healthy cell membranes, producing quenched fluorescence. When the membrane depolarizes, subsequent ion permeation releases the dye from the membrane and increases fluorescence intensity. 3The assay was performed according to a modified protocol reported previously. Briefly, the corresponding bacteria were collected from mid-logarithmic cultures by centrifugation (7000 rpm, 10 minutes) and washed three times with PBS containing 200 mM glucose. The pellet was then resuspended in the same buffer and diluted to an optical density of 0.3. The probe was then added to the suspension to a final concentration of 1 μM and inoculated into a 96-well plate at 100 μl per well. The plate was incubated in the dark for 30 minutes, and the signal was measured every 5 minutes until a stable baseline was observed. Different concentrations of COE were then added, and the change in fluorescence intensity over time was measured at 37°C. The excitation and emission wavelengths used were 600 nm and 660 nm, respectively. Quantitative membrane depolarization was induced by treating the cells with 0.2% (v / v) Triton-X-100 to completely lyse the cells, as a positive control. Independent wells containing untreated cells were used as negative controls. In the absence of bacteria, no interaction between the probe and the compound was observed. The data are plotted as the mean ± SEM of two experiments performed in triplicate.

[0294] Propidium iodide influx assay

[0295] The assay was performed according to a modified reported protocol. 4 Briefly, the corresponding bacteria were collected from mid-logarithmic cultures by centrifugation (7000 rpm, 10 minutes) and washed three times with PBS. The pellet was then resuspended in the same buffer and diluted to an optical density of 2 by extrapolation. The desired final concentration was used in a 96-well plate. Cells were treated with COE and incubated in the dark for 30 minutes. An equal volume of propidium iodide (PI) was added to the wells and mixed (final concentration was 30 μM). Fluorescence intensity was measured over time in a plate reader at an excitation wavelength of 525 nm and an emission wavelength of 620 nm. Fluorescence signal was measured every 30 seconds, repeated three times.

[0296] live-dead staining

[0297] Use LIVE / DEAD according to the manufacturer's protocol. TM BacLight TM The cell membrane integrity was determined using the Bacterial Viability Kit L7007 (ThermoFisher, USA). Bacterial cells were collected from mid-logarithmic cultures by centrifugation (7000 rpm, 10 min). The culture medium was removed, the bacteria were washed twice with PBS, and diluted to approximately 1 × 10 cells / ml. 7The concentration of CFU was calculated. The bacterial suspension was treated with DM6P to a final concentration of 2×MIC and incubated at 37°C and 200 rpm for 2 hours. A mixture of SYTO-9 and PI dyes dissolved in DMSO was added to the suspension to a final concentration of 5 μM and 30 μM, respectively. The bacteria were then incubated in the dark for 15 minutes. Samples were prepared by aliquoting 5 μL onto a clean glass slide, captured with an 18 mm coverslip, and imaged on an epifluorescence microscope (Leica Thunder Imager).

[0298] Transmission electron microscopy

[0299] The mid-logarithmic phase culture of MRSA was processed 30 minutes with 4 μ g / mL DM6P or PBS (control) respectively. Cell was fixed overnight with 4% glutaraldehyde at 4 ℃, and collected by centrifugation (7000rpm, 10 minutes). Cell was fixed after 0.2% osmic acid, ethanol gradient dehydration, and embedded in epoxy resin. Ultrathin section was placed on copper net through uranyl acetate staining, and imaged on JEOLJEM-1400Flash TEM.

[0300] Flow cytometry

[0301] Bacterial cells were collected from mid-logarithmic cultures by centrifugation (7000 rpm, 10 minutes) and washed twice with PBS. The cells were treated with 4 × MIC concentration COE for 30 minutes. PI (final concentration 30 μM) was then added and the cells were incubated for a further 10 minutes. The suspension was diluted to a suitable cell density in PBS and flow cytometry analysis was performed using an Amnis ImageStreamX Mk II imaging flow cytometer. The area-aspect ratio of the bright field channel was gated to screen out only intact cell events. A 638 nm laser was used to excite and collect emission light from 642 nm to 745 nm to select cells stained with PI. Data were processed using IDEAS v6.3.

[0302] In vivo experiments

[0303] The animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of the National University of Singapore (No. R20-1178) and performed in accordance with the Guide for the Care and Use of Laboratory Animals (National Institutes of Health, USA). C57BL6 female mice (weighing 18g to 20g) were selected and anesthetized with ketamine, and the hair on the back was shaved. A biopsy punch was used to make an excisional wound with a diameter of 6mm on the back. Logarithmic phase MRSA (in 5μL PBS) was inoculated on the wound. 3M Tegaderm was covered on the wound to prevent contamination. After 2 hours of infection, DM6P saline solution (500μg / mL, 0.25mg / kg) or vehicle control (saline) was added to the wound and left to dry for 10 minutes. New Tegaderm was applied to the wound. All mice were killed 24 hours after infection, and the wound tissue was harvested and placed in 1mL PBS. After homogenization, the samples were serially diluted 10-fold and plated on TSA plates. CFU were counted after incubation at 37°C for 16 hours. Significance was assessed using one-way analysis of variance (ANOVA) followed by multiple comparisons with one-way Tukey's test (GraphPad PRISM).

[0304] For histological analysis, a second group of mice was treated in a similar manner and monitored for 14 days after treatment. At the end point, the mice were sacrificed and organs and blood samples were collected. Tissues were fixed in 10% neutral buffered formalin (NBF) for 48 hours and stained with H&E and examined. Clinical blood biomarkers were detected using the cM4 blood biochemistry analyzer (Tianjin MNCHIP Technologies Co., Ltd.) according to the instructions.

[0305] Animal studies

[0306] To test for Staphylococcus pseudintermedius, C57BL / 6[SPF] mice (female, 18g to 20g) were first adapted to feeding, then weighed and divided into 3 groups according to body weight. After anesthesia, the mice were shaved of their back hair, and excisional wounds with a diameter of 6mm were made on the back, and the logarithmic phase Staphylococcus pseudintermedius (Sp strain) was inoculated (in 5μL PBS). 3M Tegaderm was covered on the wound to prevent contamination. After 2 hours of infection, each group was used to test saline, DM6P (0.1mg / kg) and DM6P (0.25mg / kg) by adding directly to the wound. Sampling and colony counting were performed 12 hours and 24 hours after infection. All mice were killed 24 hours after infection and the wound tissue was harvested into 1mL PBS.

[0307] It will be understood that many further modifications and permutations of the various aspects of the described embodiments are possible. Therefore, the described aspects are intended to embrace all such changes, modifications and variations that fall within the spirit and scope of the appended claims.

[0308] Throughout this specification and claims, unless the context requires otherwise, the terms "comprises" or "comprising" and variations thereof will be understood to imply the inclusion of a stated integer or step or group of integers or groups of steps but not the exclusion of any other integer or step or group of integers or groups of steps.

[0309] Throughout this specification and claims, unless the context requires otherwise, the phrase "consisting essentially of" and variations thereof will be understood to indicate that one or more of the recited elements are essential, i.e., essential elements of the invention. The phrase permits the presence of additional, unrecited elements that do not materially affect the characteristics of the invention, but excludes additional, unspecified elements that would affect the basic and novel characteristics of the defined method.

[0310] The reference in this specification to any prior art publication (or its derivative information) or to any publicly known matter shall not be regarded as an admission or any form of indication that the prior art publication or (its derivative information) or publicly known matter constitutes part of the common general knowledge in the technical field to which this specification belongs.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt, solvate or isomer thereof: in Each of R1, R2 and R3 is independently selected from H, optionally substituted alkyl, optionally substituted alkoxy; wherein at least one of R1, R2 and R3 on each phenylene ring is or wherein each R4 is independently selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, and optionally substituted cycloalkenyl; n is an integer selected from 2 to 10; and X is a linking group configured to maintain the π-π conjugation between the two phenylene rings.

2. The compound according to claim 1, wherein X is selected from in represents a bond to any phenylene ring.

3. A compound of formula (IIa) or a pharmaceutically acceptable salt, solvate or isomer thereof: in Each R1 is independently selected from H, optionally substituted alkyl, and optionally substituted alkoxy; Each R2 is independently selected from H, optionally substituted alkyl, and optionally substituted alkoxy; each R3 is independently selected from optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, and optionally substituted cycloalkenyl; and n is an integer selected from 4 to 10.

4. The compound of any one of claims 1 to 3, wherein R1 is independently optionally substituted methoxy and R2 is H.

5. The compound according to any one of claims 1 to 4, wherein at least one R4 is optionally substituted aryl.

6. The compound according to any one of claims 1 to 5, wherein the compound of formula (I) or the compound of formula (IIa) is characterized by trans configuration or E configuration.

7. The compound according to any one of claims 1 to 6, wherein the compound of formula (I) or the compound of formula (IIa) is selected from:

8. The compound of any one of claims 1 to 7, wherein the compound of formula (I) or the compound of formula (IIa) is characterized by a solubility in aqueous media of about 2 μg / mL to about 200 μg / mL, or preferably about 4 μg / mL to about 138 μg / mL.

9. The compound of any one of claims 1 to 8, wherein the compound of formula (I) or the compound of formula (IIa) is characterized by a minimum inhibitory concentration (MIC) against microorganisms of about 0.5 μg / mL to about 64 μg / mL.

10. The compound of any one of claims 1 to 9, wherein the compound of formula (I) or the compound of formula (IIa) is characterized by a minimum inhibitory concentration (MIC) against Gram-negative bacteria of about 0.5 μg / mL to about 64 μg / mL.

11. The compound according to any one of claims 1 to 10, wherein the minimum inhibitory concentration (MIC) of the compound of formula (I) or the compound of formula (IIa) against Gram-positive bacteria is about 0.5 μg / mL to about 16 μg / mL.

12. The compound of claim 11, wherein the MIC is not affected by the presence of proteins and / or salts in aqueous media.

13. The compound according to any one of claims 1 to 12, wherein the compound of formula (I) or the compound of formula (IIa) is characterized by an IC of 50 From about 5 μg / mL to about 256 μg / mL.

14. A compound according to any one of claims 1 to 13, wherein the compound of formula (I) or the compound of formula (IIa) is characterized by a selectivity index (IC 50 / MIC) is from about 2 to about 100.

15. The compound of any one of claims 1 to 14, wherein the compound of formula (I) or the compound of formula (IIa) is characterized by a minimum bactericidal concentration (MBC) of about 0.5 μg / mL to 64 μg / mL.

16. The compound according to any one of claims 1 to 15, wherein microorganisms contacted with the compound of formula (I) or the compound of formula (IIa) do not develop drug resistance.

17. The compound according to any one of claims 1 to 16, wherein the bacterial membrane of the bacterial cell contacted with the compound of formula (I) or the compound of formula (IIa) is depolarized.

18. A pharmaceutical composition comprising a compound of formula (I) or a compound of formula (IIa) or a pharmaceutically acceptable salt, solvate or isomer thereof, optionally in combination with an excipient.

19. A composition comprising a compound of formula (I) or a compound of formula (IIa) or a salt, solvate or isomer thereof, optionally in combination with an excipient.

20. A method for treating bacterial infection or bacterial disease, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I) or a compound of formula (IIa) or a pharmaceutically acceptable salt, solvate or isomer thereof.

21. A compound of formula (I) or formula (IIa) or a pharmaceutically acceptable salt, solvate or isomer thereof for use in treating bacterial infection or bacterial disease.

22. Use of a compound of formula (I) or a compound of formula (IIa) or a pharmaceutically acceptable salt, solvate or isomer thereof in the preparation of a medicament for treating bacterial infection or bacterial disease.

23. The method, compound for use or use according to any one of claims 20 to 22, wherein about 0.05 mg / kg to about 10 mg / kg of a compound of formula (I) or a compound of formula (IIa) or a pharmaceutically acceptable salt, solvate or isomer thereof is provided to a subject in need thereof.

24. The method, compound for use or use according to any one of claims 20 to 23, wherein the bacteria in the bacterial infection or bacterial disease are antibiotic-resistant bacteria.

25. The method, compound for use or use according to any one of claims 20 to 24, wherein the bacteria in the bacterial infection or bacterial disease are Gram-negative or Gram-positive bacteria.

26. The method, compound for use or use according to any one of claims 20 to 25, wherein the bacterium is selected from Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium and non-tuberculous mycobacteria.

27. The method, compound for use or use according to any one of claims 20 to 26, wherein the bacterial infection or bacterial disease is a bacterial skin infection or a foodborne infection.

28. A method of disinfecting a surface comprising contacting a compound of formula (I) and / or a compound of formula (IIa) with said surface.

29. The method of claim 28, wherein the surface is an abiotic surface.

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