Bacterial efflux pump inhibitors and methods of use

Abstract

This invention discloses bacterial efflux inhibitor compounds and methods for using such compounds to treat bacterial infections.

Description

Prior Technology

[0001] The rise of antibiotic resistance underscores the need for novel antimicrobial agents. While historical approaches to antibiotic discovery have yielded many crucial therapeutic agents, recent attempts to identify novel drugs have lagged far behind the proliferation of resistance. During the golden age of antibiotic discovery in the 1940s and 1950s, screening actinomycete extracts for growth inhibition of pathogenic bacteria was crucial. This empirical platform led to the identification of the major classes of antibiotics currently in use. This culture-based strategy resulted in inhibitors targeting nuclear growth processes: translation, DNA replication, and cell wall synthesis. While highly effective, targeting essential processes resulted in strong selection against resistance. To focus on searching for antimicrobial compounds that are less likely to contribute to resistance, the field has shifted towards screening for virulence-specific processes, aided by the advent of genomics and the co-identification of virulence-related targets. Pharmaceutical companies have invested in high-throughput screening of libraries of synthetic chemicals with inhibitory activity against validated molecular targets in biochemical analysis. Over the past 30 years, targeted approaches have produced zero antibiotics for systemic use, due to a combination of poor hit recognition from self-screening and a general lack of antibacterial activity throughout the bacteria.

[0002] The disconnect between biochemical inhibition and antibacterial activity is attributed to the intracellular accumulation of small molecules in bacteria. Specifically, Gram-negative bacteria contain a cell membrane, cell wall, and outer membrane. This cell membrane restricts the penetration of amphiphilic and hydrophilic substances into the cytoplasm and poses a significant challenge to antibiotics.

[0003] Bacteria also use efflux pumps as a mechanism for antibiotic resistance. Efflux pumps cross the periplasm between the inner and outer membranes, capturing antibiotics and host antimicrobial peptides (AMPs) and exporting them in an energy-dependent manner. When bacteria face toxic molecules such as antibiotics, they exhibit a higher level of efflux pump response. Efflux pumps capture and expel antibiotics, and most antimicrobial resistance (AMR) clinical isolates have acquired additional copies of efflux pumps and / or expressed them at high levels. Therefore, bacterial efflux pumps are a major contributor to increased antibiotic resistance in Gram-negative bacteria.

[0004] Furthermore, bacteria living within host cells (e.g., *Salmonella enterica*, *Listeria monocytogenes*, *Staphylococcus aureus*, *Mycobacterium tuberculosis*) are additionally protected by the host cell membrane; some pathogens living within vesicles are also protected by the phagosome membrane. Therefore, even traditional antibiotics used to combat extracellular pathogens are ineffective against intracellular microorganisms. For example, aminoglycosides and β-lactams accumulate poorly within host cells and are generally ineffective. Fluoroquinolones are primarily localized in the host cytoplasm and are therefore less effective against pathogens within phagosomes. Macrolides, although highly concentrated intracellularly, are generally ineffective against vesicular microorganisms because phagolysosomes are inactivated at low pH levels, as biochemical methods inherently neglect cell permeability during initial screening. Therefore, poor cell permeability represents a key deficiency for antimicrobial agents targeting virulence.

[0005] This invention addresses these needs. Summary of the Invention

[0006] In the embodiments, the present invention provides a compound of formula (I). Or its stereoisomers or pharmaceutically acceptable salts, in: n is 1, 2, or 3; p is 1, 2, 3, 4, or 5; m is 1, 2, 3, 4, or 5; Each R1 is independently a halogroup, alkyl group, or haloalkyl group; R2 is -H, alkyl, alkenyl, or alkynyl; R3 is -H, alkyl, alkenyl, or alkynyl; R4 is an alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclic group, each of which may be substituted by one or more R5 groups as appropriate; Alternatively, R3 and R4 may form a heterocyclic group, which may be substituted by one or more R5 groups, depending on the situation. R5 is aryl, heteroaryl, alkyl, NH2, NHR A or NRARB, or alkyl-NH2; RA is alkyl, alkenyl, or alkynyl, each of which may be substituted with -OH or alkoxy groups as appropriate; and RB can be alkyl, alkenyl, or ynyl.

[0007] In the embodiments of formula (I), R3 is -H or an alkyl group. In the embodiments of formula (I), R4 is an alkyl group substituted with one R5, or a heterocyclic group; and R5 is NH2. In the embodiments, the compound of formula (I) has the following structure: , Or, or a medicinally acceptable salt.

[0008] In the embodiment of formula (I), R3 and R4 together form a heterocyclic group, which may be substituted by one or more R5 groups. In the embodiment, the compound of formula (I) has the structure of formula (II): Or its stereoisomers or pharmaceutically acceptable salts, in: n is 1, 2, or 3; p is 1, 2, 3, 4, or 5; m is 1, 2, 3, 4, or 5; q is 1, 2, or 3; Each R1 is independently a halogroup, alkyl group, or haloalkyl group; R2 is hydrogen, C1-6 alkyl, C1-6 alkenyl, or C1-6 ynyl; Ring A is a heterocyclic group; R5 is aryl, heteroaryl, alkyl, NH2, NHR A, or NRARB; RA is a C1-6 alkyl, C1-6 alkenyl, or C1-6 alkynyl group, each of which may be substituted with -OH or C1-6 alkoxy groups as appropriate; and RB is a C1-6 alkyl, C1-6 alkenyl, or C1-6 alkynyl group, each of which may be substituted with -OH or C1-6 alkoxy groups as appropriate.

[0009] In the embodiments of formula (II), ring A is a 5- to 8-membered heterocyclic group, which, in addition to ring N shown in formula (II), may have 1, 2, or 3 heteroatoms selected from N, O, or S. In the embodiments, R5 is a heteroaryl, NH2, NHR A, or NRARB; RA is a C1-6 alkyl group, which may be substituted with a C1-6 alkoxy group; and RB is a C1-6 alkyl group.

[0010] In embodiments of formula (I) or (II), the compounds have the structure of formula (III): (III) Or its stereoisomers or pharmaceutically acceptable salts, in: n is 1 or 2; p is 1, 2, or 3; m is 1, 2, or 3; q is 1; Each R1 is independently a halogroup or a haloalkyl group; Ring A is a 5- or 6-membered heterocyclic group; R5 is a heteroaryl group, a C1-6 alkyl group substituted with -NH2, -NH2, -NHRA, or -NRARB; RA is a C1-6 alkyl group substituted with a C1-6 alkoxy group, as appropriate; and RB is a C1-6 alkyl group.

[0011] In the embodiments of formula (I), (II) or (III), the compounds have the structure of formula (III-1) or (III-2): Or its stereoisomers or pharmaceutically acceptable salts, The n, p, m, q, R1, A ring and R5 series are defined as above.

[0012] In this embodiment, n is 1 or 2. In this embodiment, n is 2.

[0013] In the embodiments, p is 1, 2, or 3. In the embodiments, p is 1.

[0014] In the embodiments, m is 1, 2, or 3. In the embodiments, m is 1.

[0015] In this embodiment, q is 1.

[0016] In the embodiments, each R1 is independently a halogroup or a haloalkyl group. In the embodiments, each R1 is independently a halogroup. In the embodiments, n is 2 and each R1 is independently a halogroup. In the embodiments, each R1 is Cl. In the embodiments, n is 2 and each R1 is -Cl.

[0017] In the embodiments, ring A is a 5- or 6-membered heterocyclic group.

[0018] R5 is a heteroaryl, C1-6 alkyl-NH2, -NH2, -NHRA, or -NRARB. In the examples, RA is a C1-6 alkyl group, which may be substituted with a C1-6 alkoxy group as appropriate. In the examples, RB is a C1-6 alkyl group.

[0019] In the embodiments of formula (I), (II) or (III), the compounds have the structure of formula (IV): (IV) Or its stereoisomers or pharmaceutically acceptable salts, in: n is 1 or 2; q is 1; Each R1 is a Cl or fluoroalkyl group; R5 is an alkyl group substituted with -NH2, or -NH2, -NHRA, or NRARB; RA is a C1-6 alkyl group substituted with a C1-6 alkoxy group, as appropriate; RB is a C1-6 alkyl group.

[0020] In the embodiments, the compounds of formulas (I), (II), (III) or (IV) have the structure of formula (IV-1) or (IV-2): Or its stereoisomers or pharmaceutically acceptable salts, Where n, q, R1 and R5 are as defined above.

[0021] In the examples, R5 is an alkyl group substituted with -NH2, or -NH2. In the examples, the compound of formula (IV) has the following structure: , Or, its stereoisomers or pharmaceutically acceptable salts. In the examples, the compound of formula (IV) has the following structure: Or, or a medicinally acceptable salt.

[0022] In the embodiments of formula (I), (II) or (III), the compounds have the structure of formula (V): Or its stereoisomers or pharmaceutically acceptable salts, in: n is 1 or 2; q is 1; Each R1 is a Cl or fluoroalkyl group; R5 is a heteroaryl, NH2, NHRA, or NRARB; RA is a C1-6 alkyl group substituted with a C1-6 alkoxy group, as appropriate; RB is a C1-6 alkyl group.

[0023] In the embodiments of formula (V), R5 is a 5- to 7-membered heteroaryl group having 1, 2, or 3 heteroatoms selected from N and S. In the embodiments, R5 is a 5-membered heteroaryl group having 1 or 2 N heteroatoms. In the embodiments, R5 is an imidazole group. In the embodiments, the compound of formula (V) has the following structure: Or a medicinally acceptable salt.

[0024] In the embodiments, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I), (II), (III), (IV) or (V) and one or more pharmaceutically acceptable excipients.

[0025] In an embodiment, the present invention provides a method for treating a bacterial infection in an individual in need, comprising administering to the individual a pharmaceutically acceptable amount of a compound of formula (I), (II), (III), (IV), or (V). In an embodiment, the bacterial infection is caused by an intracellular pathogen. In an embodiment, the bacterial infection is caused by Gram-negative bacteria.In the embodiments, the bacterial infection is caused by one or more of the following genera: Salmonella (sp.), Acinetobacter (sp.), Actinobacillus (sp.), Aeromonas (sp.), Bacteroides (sp.), Bordetellus (sp.), Brucella (sp.), Burkholderia (sp.), Prevotellus (sp.), Porphyromonas (sp.), Campylobacter (sp.), Citrobacter (sp.), Edwarsiella (sp.), Eikenell (sp.), Enterobacter (sp.), Escherichia (sp.), Francisella (sp.), Haemophilus (sp.), and Helicobacter (sp.). Helicobacter p.), Kingella sp., Klebsiella sp., Legionella sp., Moraxella sp., Morganella sp., Neisseria sp., Pasteurell sp., Plesiomonas sp., Proteus sp., Providenciasp., Pseudomonas sp., Serratia sp., Shigella sp., Stenotrophomonas sp., Streptobacillus sp., Vibrio sp., Yersini sp., Chlamydophila sp., Rickettsia sp. Caused by *Rhizoctonia* sp., *Coxiella* sp., *Ehrlichiasp.*, or *Bartonella* sp. In the examples, the bacterial infection was caused by one or more *Salmonella* species.In the examples, the Salmonella species is *Enterobacter* (a serovar of *Salmonella typhimurium*). In the examples, the bacterial infection is caused by *Escherichia coli*, *Klebsiella pneumoniae*, or *Enterobacter cloacae*. In the examples, the bacterial infection is resistant to one or more antibiotics. In the examples, the method of the present invention further includes administering one or more antibiotics. In the examples, the antibiotic is a macrocyclic lactone, tetracycline, fluoroquinolone, penicillin, cephalosporin, aminoglycoside, sulfonamide, β-lactam, trimethoprim-sulfamethoxazole, chloramphenicol, or lincosamide.

[0026] In an embodiment, the present invention provides a method for inhibiting bacterial efflux pumps in an individual with a bacterial infection, comprising administering to the individual a pharmaceutically acceptable amount of a compound of formula (I), (II), (III), (IV), or (V). In an embodiment, the bacterial infection is caused by an intracellular pathogen. In an embodiment, the bacterial infection is caused by Gram-negative bacteria. In an embodiment, the bacterial infection is caused by one or more of the following bacteria: Salmonella spp., Acinetobacter spp., Actinobacter spp., Aeromonas spp., Bacteroides spp., Bodophyllum spp., Brucella spp., Burkholderia spp., Prevotella spp., Porphyromonas spp., Campylobacter spp., Citrobacter spp., Edwardsiella spp., Ekenella spp., Enterobacter spp., Escherichia spp., Franz spp., Haemophilus spp., Helicobacter spp., and Chlorella spp. The infection is caused by one or more Salmonella species. In one example, the Salmonella spp. includes Klebsiella, Legionella, Moraxella, Morganella, Neisseria, Pasteurella, Orthomonas, Proteus, Providencia, Pseudomonas, Serratia, Shigella, Oligotrophozoites, Streptococcus, Vibrio, Yersinia, Chlamydia, Rickettsia, Coxiella, Ehrlich, or Bartonella. In the examples, the bacterial infection is caused by one or more Salmonella species. In one example, the Salmonella spp. is Enterobacter (a serovar of Salmonella typhimurium). In one example, the bacterial infection is caused by Escherichia coli, Klebsiella pneumoniae, or Enterobacter cloacae. In one example, the bacterial infection is resistant to one or more antibiotics. In one example, the method of the present invention further includes administering one or more antibiotics. In the examples, the antibiotic is a macrocyclic lactone, tetracycline, fluoroquinolone, penicillin, cephalosporin, aminoglycoside, sulfonamide, β-lactam, trimeprine-sulfamethoxazole, chloramphenicol, or lincosamide.

[0027] In the embodiments, the present invention provides a method for increasing the sensitivity of Gram-negative bacteria to antibiotics, which includes combining a compound of formula (I), (II), (III), (IV) or (V) with an antibiotic.

[0028] In the embodiments, the present invention provides a method for reversing or reducing antibiotic resistance in antibiotic-resistant Gram-negative bacteria, comprising administering a compound of formula (I), (II), (III), (IV) or (V). Implementation

[0029] [Cross-reference to related applications] []

[0030] This application claims priority to U.S. Application No. 63 / 228,541, filed August 2, 2021. The entire contents of the aforementioned application are incorporated herein by reference. [Government Support] []

[0031] This invention was carried out with government support and under grant NIH R33 AI121365 granted by the National Institutes of Health. The government retains certain rights to this invention. [definition] []

[0032] The term "medically acceptable salt" includes both acid and base addition salts. Pharmaceutically acceptable salts include those obtained by reacting an active compound acting as a base with an inorganic or organic acid to form a salt, such as salts of hydrochloric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, camphorsulfonic acid, oxalic acid, maleic acid, succinic acid, citric acid, formic acid, hydrobromic acid, benzoic acid, tartaric acid, fumaric acid, salicylic acid, mandelic acid, and carbonic acid. Those skilled in this art will further appreciate that acid addition salts can be prepared by reacting a compound with a suitable inorganic or organic acid via any of many known methods.

[0033] The term "treatment" means one or more of the following: reducing, alleviating, delaying, decreasing, improving, or managing at least one symptom of an individual's condition. The term "treatment" may also mean one or more of the following: inhibiting the development or worsening of a condition, delaying the onset of a condition (i.e., prior to the clinical manifestation of the condition), or reducing the risk of developing or worsening a condition.

[0034] The compounds of this invention, or their pharmaceutically acceptable salts, contain at least one asymmetric center. Compounds of this invention having one asymmetric center produce enantiomers, wherein absolute stereochemistry can be represented as (R)- and (S)-, or (+) and (-). When compounds of this invention have more than two asymmetric centers, such compounds may exist as diastereomers or other stereoisomers. This invention is intended to encompass all such possible isomers, as well as their racemic and, where appropriate, pure forms, whether or not explicitly described herein. Optically active (+) and (-) or (R)- and (S)- isomers can be prepared using palmitic synthons or palmitic reagents, or resolved using known techniques (e.g., chromatography and fractional crystallization). Known techniques for the preparation / separation of individual enantiomers include resolution from suitable optically pure precursors via palmitic synthesis or racemic synthesis (or racemic derivatives of salts or derivatives) using, for example, palmitic high-performance liquid chromatography (HPLC). When the compounds described herein contain an olefinic double bond or other geometrically asymmetric center, and unless otherwise specified, it is intended that such compounds include both E and Z geometric isomers. Similarly, it is intended that all tautomer forms be included.

[0035] "Stereoisomers" refer to compounds composed of identical atoms with the same bonds but different three-dimensional structures, and these structures are not interchangeable. This invention covers various stereoisomers and mixtures thereof, including "enantiomers," which refer to two stereoisomers whose molecules are non-superimposed mirror images of each other.

[0036] The term "therapeuticly effective" when applied to dosage or amount refers to the quantity of a compound or pharmaceutical preparation that, when administered to a patient in need, is sufficient to produce the desired clinical benefit.

[0037] The term "halogen" refers to halogens. Specifically, it refers to fluorine, chlorine, bromine, and iodine.

[0038] "Alkyl / alkyl group" refers to a fully saturated straight-chain or branched hydrocarbon chain group connected to the rest of the molecule by a single bond. It includes alkyl groups containing any number of carbon atoms (including but not limited to 1 to 12). Alkyl groups containing up to 12 carbon atoms are C1-C12 alkyl groups, alkyl groups containing up to 10 carbon atoms are C1-C10 alkyl groups, alkyl groups containing up to 6 carbon atoms are C1-C6 alkyl groups, and alkyl groups containing up to 5 carbon atoms are C1-C5 alkyl groups. C1-C5 alkyl groups include C5 alkyl, C4 alkyl, C3 alkyl, C2 alkyl, and C1 alkyl (i.e., methyl). C1-C6 alkyl groups include not only all the portions described above for C1-C5 alkyl groups, but also C6 alkyl groups. C1-C10 alkyl groups include not only all the portions described above for C1-C5 and C1-C6 alkyl groups, but also C7, C8, C9, and C10 alkyl groups. Similarly, C1-C12 alkyl groups include not only all the portions described above, but also C11 and C12 alkyl groups. Non-limiting examples of C1-C12 alkyl groups include methyl, ethyl, n-propyl, isopropyl, dipropyl, n-butyl, isobutyl, dibutyl, tributyl, n-pentyl, tripentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, and n-dodecyl. Unless otherwise specified, alkyl groups may be substituted as appropriate.

[0039] "Alkenyl (alkenyl group)" refers to a straight-chain or branched hydrocarbon chain having 2 to 12 carbon atoms and one or more carbon-carbon double bonds. Each alkenyl group is connected to the rest of the molecule by a single bond. It includes alkenyl groups containing any number of carbon atoms from 2 to 12. Alkenyl groups containing up to 12 carbon atoms are C2-C12 alkenyl groups, alkenyl groups containing up to 10 carbon atoms are C2-C10 alkenyl groups, alkenyl groups containing up to 6 carbon atoms are C2-C6 alkenyl groups, and alkenyl groups containing up to 5 carbon atoms are C2-C5 alkenyl groups. C2-C5 alkenyl groups include C5 alkenyl, C4 alkenyl, C3 alkenyl, and C2 alkenyl groups. C2-C6 alkenyl groups include not only all the portions described above for C2-C5 alkenyl groups but also C6 alkenyl groups. The C2-C10 alkenyl group includes not only all the portions described above for the C2-C5 and C2-C6 alkenyl groups, but also C7, C8, C9, and C10 alkenyl groups. Similarly, the C2-C12 alkenyl group includes not only all the portions described above, but also C11 and C12 alkenyl groups. Non-limiting examples of 12-alkenyl groups include ethenyl, 1-propenyl, 2-propenyl (allyl), isopropenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 5-heptenyl, 6-heptenyl, 1-octenyl, 2-octenyl, 3-octenyl, 4-octenyl, 5-octenyl, 6-octenyl, 7-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl... 4-Nonenyl, 5-Nonenyl, 6-Nonenyl, 7-Nonenyl, 8-Nonenyl, 1-Decanyl, 2-Decanyl, 3-Decanyl, 4-Decanyl, 5-Decanyl, 6-Decanyl, 7-Decanyl, 8-Decanyl, 9-Decanyl, 1-Undecenyl, 2-Undecenyl, 3-Undecenyl, 4-Undecenyl, 5-Undecenyl, 6-Undecenyl, 7-Undecenyl, 8-Undecenyl, 9-Undecenyl, 10-Undecenyl, 1-Dodecenyl, 2-Dodecenyl, 3-Dodecenyl, 4-Dodecenyl, 5-Dodecenyl, 6-Dodecenyl, 7-Dodecenyl, 8-Dodecenyl, 9-Dodecenyl, 10-Dodecenyl, and 11-Dodecenyl. Unless otherwise specified, the alkenyl group may be substituted as appropriate.

[0040] "Alkynyl / alkynyl group" refers to a straight-chain or branched hydrocarbon chain having 2 to 12 carbon atoms and one or more carbon-carbon triple bonds. Each alkynyl group is connected to the rest of the molecule by a single bond. It includes alkynyl groups containing any number of carbon atoms from 2 to 12. Alkynyl groups containing up to 12 carbon atoms are C2-C12 alkynyl groups, those containing up to 10 carbon atoms are C2-C10 alkynyl groups, those containing up to 6 carbon atoms are C2-C6 alkynyl groups, and those containing up to 5 carbon atoms are C2-C5 alkynyl groups. C2-C5 alkynyl groups include C5 alkynyl, C4 alkynyl, C3 alkynyl, and C2 alkynyl groups. C2-C6 alkynyl groups include not only all the portions described above for C2-C5 alkynyl groups but also C6 alkynyl groups. The C2-C10 ynyl group includes not only all the portions described above for the C2-C5 and C2-C6 ynyl groups, but also C7, C8, C9, and C10 ynyl groups. Similarly, the C2-C12 ynyl group includes not only all the portions described above, but also C11 and C12 ynyl groups. Non-limiting examples of the C2-C12 ynyl group include ethynyl, propynyl, butynyl, pentynyl, and the like. Unless otherwise specified, the ynyl group may be substituted as appropriate.

[0041] "Alkoxy" refers to a group of the formula -OR a, where Ra is an alkyl, alkenyl, or alkynyl group containing 1 to 12 carbon atoms as defined above. Unless otherwise specified, alkoxy groups may be substituted as appropriate.

[0042] "Cycloalkyl" refers to a stable, non-aromatic, monocyclic or polycyclic, fully saturated hydrocarbon group consisting only of carbon and hydrogen atoms. It may contain a fused or bridged ring system having 3 to 20 carbon atoms, preferably 3 to 10 carbon atoms, and is connected to the rest of the molecule by a single bond. Monocyclic cycloalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyl groups include, for example, adamantyl, norbornyl, decahydronaphthyl, 7,7-dimethyl-bicyclo[2.2.1]heptyl, and the like. Unless otherwise expressly specified in this specification, cycloalkyl groups may be substituted as appropriate.

[0043] "Halogenalkyl" means an alkyl group as defined above that is substituted with one or more halogen groups as defined above, such as trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl and the like. Unless otherwise specified, halogenalkyl groups may be substituted as appropriate.

[0044] "Aryl" refers to a hydrocarbon ring system group comprising hydrogen, 6 to 18 carbon atoms, and at least one aromatic ring. For the purposes of this invention, aryl can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems. Aryl includes (but is not limited to) aryl derivatives of: anthracene, phenanthrene, anthracene, azulene, benzene, arsenic, arsenic, arsenic, asymmetric indole, symmetric indole, dihydroindene, indene, naphthalene, phenanthracene, heptamethrin, pyrene, and benzo[a]phenanthrene. Unless otherwise specified, the term "aryl" is intended to include, where appropriate, substituted aryl groups.

[0045] "Heterocyclic group," "heterocyclic ring," or "heterocycle" refers to a stable 3- to 20-membered cyclic group consisting of 2 to 12 carbon atoms and 1 to 6 heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur. The heterocyclic group can be a monocyclic, bicyclic, tricyclic, or tetracyclic system, and may include fused or bridging ring systems. The nitrogen, carbon, or sulfur atoms in the heterocyclic group may be oxidized, and the nitrogen atom may be quaternized, depending on the situation. The heterocyclic group may be partially or completely saturated. Examples of such heterocyclic groups include (but are not limited to) dioxolane, decahydroisoquinolinyl, imidazolinyl, imidazodinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-sideoxypiperazinyl, 2-sideoxypiperidinyl, 2-sideoxypyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidinoneyl, pyrrolidinyl, pyrazolidinyl, quininecycloyl, thiazodinyl, tetrahydrofuranyl, trithiaalkyl, tetrahydropiperazinyl, thiomorpholinyl, thiomorpholinyl, 1-sideoxy-thiomorpholinyl, and 1,1-disideoxy-thiomorpholinyl. Unless otherwise specified, heterocyclic groups may be substituted as appropriate.

[0046] "Heteroaryl" refers to a 5- to 20-membered ring system group comprising a hydrogen atom, 1 to 13 carbon atoms, 1 to 6 heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur, and at least one aromatic ring. For the purposes of this invention, the heteroaryl group can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon, or sulfur atoms in the heteroaryl group may be oxidized as appropriate; the nitrogen atom may be quaternized as appropriate. Examples include (but are not limited to) nitrogen-containing heterocyclic heptadienyl, acridinel, benzimidazolyl, benzothiazolyl, benzoindolyl, benzom-dioxacyclopentenyl, benzofuranyl, benzoxazolyl, benzothiadiazolyl, benzo[ b][1,4]dioxaneheptyl, 1,4-benzodioxyl, benzonaphthofuranyl, benzoxazolylbenzodioxinyl, benzopiperanyl, benzopiperanone, benzofuranyl, benzofuranone, benzothienyl / benzothiophenyl, benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridyl, carbazole, cenyl, dibenzofuranyl 1-Franyl, dibenzothiophene, furanyl, furanone, isothiazolyl, imidazolyl, indazole, indole, isoindole, indolin, isoindolin, isoquinolinyl, indazinyl, isoxazolyl, naphridinyl, oxadiazolyl, 2-side-oxy-azine-heptyl, oxazolyl, oxacyclopropane, 1-oxonylpyridinyl, 1-oxonylpyrimidinyl, 1-oxonylpyrazinyl, 1-oxonylpyrazinyl, 1-phenyl-1 H-pyrrole, phenazinyl, phenothiazinyl, phenotoxazinyl, phthalazinyl, pteridinyl, purine, pyrrole, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyrazinyl, quinazolinyl, quinoxolinyl, quinolinyl, quininecycloyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and phenylthio (i.e., thiophene). Unless otherwise specified, heteroaryl groups may be substituted as appropriate.

[0047] As used herein, the term "substitution" means any of the above groups in which at least one hydrogen atom is substituted by a bond to a non-hydrogen atom such as (but not limited to) the following: halogen atoms, such as F, Cl, Br and I; oxygen atoms in groups such as hydroxyl, alkoxy and ester groups; sulfur atoms in groups such as thiols, thioalkyls, ternaryls, sulfonyls and ternidines; nitrogen atoms in groups such as amines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides, amides and enamines; silicon atoms in groups such as trialkylsilyls, dialkylarylsilyls, alkyldiarylsilyls and triarylsilyls; and other heteroatoms in various other groups. "Substitution" also means any of the above groups in which one or more hydrogen atoms are replaced by a higher-order bond (e.g., double or triple bond) of a heteroatom such as: oxygen in a side oxygen group, carbonyl group, carboxyl group, or ester group; and nitrogen in a group such as imine, oxime, hydrazone, or nitrile. For example, "substitution" includes any of the above groups in which one or more hydrogen atoms are replaced by -NR gR h, -NR gC(=O)R h, -NR gC(=O)NR gR h, -NR gC(=O)OR h, -NR gSO 2R h, -OC(=O)NR gR h, -OR g, -SR g, -SOR g, -SO 2R g, -OSO 2R g, -SO 2OR g, =NSO 2R g, and -SO 2NR gR h. "Substitution" also means any of the above groups, wherein one or more hydrogen atoms are substituted by -C(=O)Rg, -C(=O)ORg, -C(=O)NRgRh, -CH2SO2Rg, or -CH2SO2NRgRh. In the above, Rg and Rh are the same or different and independently hydrogen, alkyl, alkenyl, alkynyl, alkoxy, alkylamine, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclic, N-heterocyclic, heterocyclic alkyl, heteroaryl, N-heteroaryl and / or heteroarylalkyl. "Substitution" further means any of the above groups in which one or more hydrogen atoms are substituted into the following bonds: amino, cyano, hydroxyl, imino, nitro, oxy-group, thio-group, halogen, alkyl, alkenyl, alkynyl, alkoxy, alkylamine, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclic, N-heterocyclic, heterocyclic alkyl, heteroaryl, N-heteroaryl and / or heteroarylalkyl. Furthermore, each of the above groups may also be substituted with one or more of the above groups as appropriate. [Compound] []

[0048] In the embodiments, the present invention provides compounds that can be used to treat bacterial infections. In the embodiments, the compounds inhibit efflux pumps, which reduce the ability of bacteria to expel antibiotics. Therefore, by inhibiting the efflux pumps, the compounds of the present invention also increase the sensitivity of bacteria to antibiotics. Therefore, in the embodiments, these compounds can be used in combination with antibiotics to treat antibiotic-resistant bacteria, reduce the ability of bacteria to develop antibiotic resistance, or increase the sensitivity of bacteria to antibiotics.

[0049] In some embodiments, the present invention provides compounds of formula (I). Or its stereoisomers or pharmaceutically acceptable salts.

[0050] In the embodiments, n is 1, 2 or 3.

[0051] In the embodiments, p is 1, 2, 3, 4 or 5.

[0052] In the embodiments, m is 1, 2, 3, 4 or 5.

[0053] In the examples, each R1 is independently a halogroup, alkyl group, or haloalkyl group. In the examples, the alkyl group is substituted. In the examples, the alkyl group is unsubstituted.

[0054] In the examples, R2 is -H, alkyl, alkenyl, or alkynyl. In the examples, the alkyl group is substituted. In the examples, the alkyl group is unsubstituted. In the examples, the alkenyl group is substituted. In the examples, the alkenyl group is unsubstituted. In the examples, the alkynyl group is substituted. In the examples, the alkynyl group is unsubstituted.

[0055] In the examples, R3 is -H, alkyl, alkenyl, or alkynyl. In the examples, the alkyl group is substituted. In the examples, the alkyl group is unsubstituted. In the examples, the alkenyl group is substituted. In the examples, the alkenyl group is unsubstituted. In the examples, the alkynyl group is substituted. In the examples, the alkynyl group is unsubstituted.

[0056] In the examples, R4 is an alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclic group, each of which may be substituted by one or more R5 groups as appropriate. In the examples, the alkyl group is substituted. In the examples, the alkyl group is unsubstituted. In the examples, the alkenyl group is substituted. In the examples, the alkenyl group is unsubstituted. In the examples, the alkynyl group is substituted. In the examples, the alkynyl group is unsubstituted. In the examples, the cycloalkyl group is substituted. In the examples, the cycloalkyl group is unsubstituted. In the examples, the heterocyclic group is substituted. In the examples, the heterocyclic group is unsubstituted.

[0057] In the embodiments, R3 and R4 together form a heterocyclic group, which may be substituted by one or more R5s as appropriate.

[0058] In the examples, R5 is an aryl, heteroaryl, alkyl, NH2, NHR A or NRARB, or an alkyl group substituted with NH2. In the examples, RA is an alkyl, alkenyl, or alkynyl group, each of which is substituted with -OH or alkoxy groups as appropriate. In the examples, RB is an alkyl, alkenyl, or alkynyl group. In the examples, the alkyl group is substituted. In the examples, the alkyl group is unsubstituted. In the examples, the alkenyl group is substituted. In the examples, the alkynyl group is unsubstituted.

[0059] In the embodiments, n is 1, 2, or 3. In the embodiments, n is 1 or 2. In the embodiments, n is 1. In the embodiments, n is 2. In the embodiments, each R1 is independently a halogroup, alkyl group, or haloalkyl group. In the embodiments, each R1 is independently a halogroup or haloalkyl group. In the embodiments, n is 2, and each R1 is independently a halogroup. In the embodiments, n is 1, and R1 is a haloalkyl group. In the embodiments, the haloalkyl group is a C1-C6 alkyl group substituted with 1, 2, 3, or more fluorine atoms. In the embodiments, the haloalkyl group is -CF3. In the embodiments, p is 1, 2, 3, 4, or 5. In the embodiments, p is 1.

[0060] In the embodiments, m is 1, 2, 3, 4, or 5. In the embodiments, m is 1.

[0061] In the embodiments, R2 is -H, alkyl, alkenyl, or alkynyl. In the embodiments, R2 is -H.

[0062] In the embodiments, R3 is -H, alkyl, alkenyl, or alkynyl. In the embodiments, R3 is -H or alkyl. In the embodiments, R3 is -H. In the embodiments, R3 is alkyl. In the embodiments, R3 is C1-C6 alkyl. In the embodiments, R3 is methyl or ethyl. In the embodiments, R3 is methyl.

[0063] In the examples, R4 is an alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclic group, each of which may be substituted with one or more R5 groups as appropriate. In the examples, R4 is an alkyl or heterocyclic group, each of which may be substituted with one or more R5 groups as appropriate. In the examples, R4 is a C1-C6 alkyl group. In the examples, R4 is methyl, ethyl, propyl (e.g., n-propyl, isopropyl, dipropyl) or butyl (e.g., n-butyl, isobutyl, dibutyl, tributyl). In the examples, R4 is propyl. In the examples, R4 is butyl. In the examples, R5 is an alkyl-NH2 group. In the examples, R5 is a C1-C6 alkyl group substituted with -NH2 (i.e., methyl, ethyl, n-propyl, isopropyl, dipropyl, n-butyl, isobutyl, dibutyl, tributyl, n-pentyl, tripentyl, or n-hexyl). In the examples, R5 is a C4 alkyl-NH2. In the examples, R5 is a C1-NH2.

[0064] In the embodiments, R4 is a 5- to 7-membered heterocyclic group having 1, 2, or 3 heteroatoms selected from O, N, or S and, if applicable, substituted with R5. In the embodiments, R4 is a 5- to 6-membered heterocyclic group. In the embodiments, R4 is a 5- to 6-membered bicyclic heterocyclic group. In the embodiments, R4 is a 6-membered heterocyclic group having 1 N heteroatom. In the embodiments, R4 is a 5,3-fused heterocyclic group having 1 N heteroatom.

[0065] In the examples, R3 is -H or alkyl (e.g., C1-3 alkyl), and R4 is alkyl or heterocyclic, which may be substituted with one R5. In the examples, R5 is -NH2. In the examples, the compounds have one of the following structures: , Or, or a medicinally acceptable salt.

[0066] In the embodiments, R3 and R4 together form a heterocyclic group. In the embodiments, the heterocyclic group may be a 5- to 8-membered heterocyclic group, which may be substituted with one or more R5s as appropriate. In the embodiments, R3 and R4 together form a 5-membered heterocyclic group, which may be substituted with -NH2 or alkyl-NH2 as appropriate. In the embodiments, R3 and R4 together form a 5-membered heterocyclic group, which may be substituted with -NH2 or C1-C6 alkyl-NH2 (e.g., -CH2-NH2). In the embodiments, R3 and R4 together form a 6-membered heterocyclic group, which may be substituted with one or more R5s as appropriate. In the embodiments, R3 and R4 together form a 6-membered heterocyclic group, which may be substituted with one or more R5s, and R5 is an alkyl, -NH2, or -NRAR B, wherein RA is an alkyl group substituted with an alkoxy group as appropriate and RB is an alkyl group. In the embodiments, R3 and R4 together form a 6-membered heterocyclic group, and R5 is a C1-C6 alkyl, -NH2, or -NRAR B, wherein RA is a C1-C6 alkyl group substituted with a C1-C6 alkoxy group, and RB is a C1-C6 alkyl group. In the embodiments, R3 and R4 together form a 6-membered heterocyclic group, which is substituted with R5, and R5 is -NH2 or -NRAR B, wherein RA is a C2-C6 alkyl group substituted with a C1-C3 alkoxy group, and RB is a C1-C3 alkyl group.

[0067] In the embodiments, R5 is an aryl, heteroaryl, alkyl, -NH2, -NHR A, or -NRAR B, or an alkyl-NH2. In the embodiments, R5 is a 6- or 8-membered aryl, a 5- to 8-membered heteroaryl, a C1-C6 alkyl, -NH2, -NHR A, or -NRAR B, or a C1-C6 alkyl substituted with -NH2. In the embodiments, R5 is a 5-membered heteroaryl. In the embodiments, R5 is an imidazolyl. In the embodiments, R3 and R4 together form a 6-membered heterocyclic group, and R5 is a 5-membered heteroaryl (e.g., imidazolyl).

[0068] In the examples, RA is alkyl, alkenyl, or alkynyl, each of which may be substituted with -OH or alkoxy groups as appropriate. In the examples, RA is a C1-C6 alkyl. In the examples, RA is methyl. In the examples, RA is ethyl. In the examples, RA is propyl. RA is a C1-C6 alkyl group substituted with a C1-C6 alkoxy group. In the examples, RA is a C1-C6 alkyl group substituted with a methoxy group. In the examples, RA is a propyl group substituted with a methoxy group.

[0069] In the examples, RB is alkyl, alkenyl, or alkynyl, each of which may be substituted with -OH or alkoxy groups as appropriate. In the examples, RB is C1-C6 alkyl. In the examples, RB is methyl.

[0070] In the embodiments, the compound of formula (I) has the structure of formula (II): Or its stereoisomers or pharmaceutically acceptable salts.

[0071] In this embodiment, n is 1, 2, or 3. In this embodiment, n is 1. In this embodiment, n is 2.

[0072] In this embodiment, p is 1, 2, 3, 4, or 5. In this embodiment, p is 1. In this embodiment, m is 1, 2, 3, 4, or 5. In this embodiment, m is 1.

[0073] In this embodiment, q is 1, 2, or 3. In this embodiment, q is 1. In this embodiment, q is 2.

[0074] In the embodiments, each R1 is independently a halogroup, alkyl group, or haloalkyl group. In the embodiments, each R1 is independently a halogroup or haloalkyl group. In the embodiments, n is 2, and each R1 is independently a halogroup. In the embodiments, n is 2, and each R1 is independently -Cl. In the embodiments, n is 1, and R1 is a haloalkyl group. In the embodiments, the haloalkyl group is a C1-C6 alkyl group substituted with 1, 2, 3, or more fluorine atoms. In the embodiments, the haloalkyl group is -CF3.

[0075] In the examples, R2 is hydrogen, C1-6 alkyl, C1-6 alkenyl, or C1-6 alkynyl. In the examples, R2 is hydrogen.

[0076] In the embodiments, ring A is a heterocyclic group. In the embodiments, ring A is a 5- to 8-membered heterocyclic group, which, in addition to ring N shown in formula (II), may have 1, 2, or 3 heteroatoms selected from N, O, or S. In the embodiments, ring A is a 5- to 8-membered heterocyclic group.

[0077] In the examples, R5 is aryl, heteroaryl, alkyl, NH2, NHR A, or NRARB. In the examples, R5 is NH2. In the examples, R5 is methyl, ethyl, or propyl (n-propyl, isopropyl, dipropyl). In the examples, R5 is methyl.

[0078] In the examples, RA is a C1-6 alkyl, C1-6 alkenyl, or C1-6 alkynyl, each of which may be substituted with a halogen, OH, or C1-6 alkoxy group, as appropriate. In the examples, RB is a C1-6 alkyl, C1-6 alkenyl, or C1-6 alkynyl, each of which may be substituted with a halogen, OH, or C1-6 alkoxy group, as appropriate. In the examples, R5 is a heteroaryl, NH2, NHR A, or NRARB. In the examples, RA is a C1-6 alkyl group substituted with a C1-6 alkoxy group, as appropriate. In the examples, RA is a propyl group substituted with a methoxy group. In the examples, RB is a C1-6 alkyl group. In the examples, RB is methyl or ethyl. In the examples, RB is methyl.

[0079] In the embodiments, the compound of formula (I) or (II) has the structure of formula (III): , (III) Or its stereoisomers or pharmaceutically acceptable salts.

[0080] In the embodiments, the compound of formula (I), (II) or (III) has the structure of formula (III-1) or (III-2): Or its stereoisomers or pharmaceutically acceptable salts.

[0081] In this embodiment, n is 1 or 2. In this embodiment, n is 2.

[0082] In the embodiments, p is 1, 2, or 3. In the embodiments, p is 1.

[0083] In the embodiments, m is 1, 2, or 3. In the embodiments, m is 1.

[0084] In this embodiment, q is 1.

[0085] In the embodiments, each R1 is independently a halogroup or a haloalkyl group. In the embodiments, each R1 is independently a halogroup. In the embodiments, n is 2 and each R1 is independently a halogroup. In the embodiments, each R1 is Cl. In the embodiments, n is 2 and each R1 is independently -Cl.

[0086] In the embodiments, ring A is a 5- or 6-membered heterocyclic group.

[0087] In the examples, R5 is a heteroaryl, C1-6 alkyl, C1-6 alkyl-NH2, -NH2, -NHRA, or -NRARB. In the examples, R5 is -NH2. In the examples, R5 is a C1-6 alkyl (e.g., -CH3) and -NH2. In the examples, RA is a C1-6 alkyl substituted with a C1-6 alkoxy group, as appropriate. In the examples, RA is a propyl group substituted with a methoxy group. In the examples, RB is a C1-6 alkyl group. In the examples, RB is a methyl group.

[0088] In the embodiments, the compound of formula (I), (II), or (III) has the structure of formula (IV): (IV) Or its stereoisomers or pharmaceutically acceptable salts.

[0089] In the embodiments, the compounds of formulas (I), (II), (III) or (IV) have the structure of formula (IV-1) or (IV-2): Or its stereoisomers or pharmaceutically acceptable salts.

[0090] In the embodiments, n is 1 or 2. In the embodiments, each R1 is Cl or a fluoroalkyl group. In the embodiments, n is 2 and each R1 is Cl. In the embodiments, n is 1 and R1 is independently -CF3.

[0091] In the examples, q is 1. In the examples, R5 is an alkyl group substituted with -NH2 or -NH2, -NHR A, or NRARB. In the examples, RA is a C1-6 alkyl group, which may be substituted with a C1-6 alkoxy group. In the examples, RB is a C1-6 alkyl group. In the examples, R5 is an alkyl group substituted with -NH2 or -NH2.

[0092] In the embodiments, the compound of formula (IV) has the structure of formula (IV.A) or (IV.B): or .

[0093] In the embodiments, the compound of formula (IV) has the following structure: , Or its stereoisomers or pharmaceutically acceptable salts.

[0094] In the embodiments, the compound of formula (IV) has the following structure: Or, or a medicinally acceptable salt.

[0095] In the embodiments, the compound of formula (I), (II), or (III) has the structure of formula (V): Or its stereoisomers or pharmaceutically acceptable salts.

[0096] In the examples, n is 1 or 2. In the examples, each R1 is Cl or fluoroalkyl. In the examples, n is 2 and each R1 is Cl. In the examples, n is 1 and R1 is fluoroalkyl. In the examples, the fluoroalkyl is a C1-C6 alkyl substituted with 1, 2, 3 or more fluorine atoms. In the examples, the fluoroalkyl is -CF3. In the examples, q is 1. In the examples, R5 is heteroaryl, NH2, NHR A or NRARB. In the examples, RA is a C1-6 alkyl substituted with C1-6 alkoxy, as appropriate. RB is a C1-6 alkyl. RA is a C1-6 alkyl substituted with C1-6 alkoxy. In the examples, RA is a C1-6 alkyl substituted with methoxy. In the examples, RA is propyl or butyl substituted with -OCH3. In the examples, RB is a C1-6 alkyl. In the examples, RB is -CH3.

[0097] In the embodiments, R5 is a 5- to 7-membered heteroaryl group having 1, 2, or 3 heteroatoms selected from N and S. In the embodiments, R5 is a 5-membered heteroaryl group having 1 or 2 N heteroatoms. In the embodiments, R5 is an imidazole group.

[0098] In the embodiments, the compound of formula (V) has the structure of formulas (VA) to (VF):

[0099] In the embodiments, the compound of formula (V) has the following structure: Or a medicinally acceptable salt. [General Synthesis] []

[0100] The compounds disclosed herein (including their salts) can be prepared using known organic synthetic techniques and can be synthesized via any of the many possible synthetic routes. Those familiar with synthetic organic chemistry will understand that the choice of starting materials and reagents will depend in part on the desired product and / or the reagents used; for example, various mechanisms require primary or secondary alcohols.

[0101] The reactions used to prepare the compounds disclosed herein can be carried out in suitable solvents, which can be readily selected by those skilled in organic synthesis. Suitable solvents are substantially non-reactive with the starting materials, intermediates, or products at the temperature at which the reaction is carried out (e.g., a temperature ranging from room temperature to the boiling point of the solvent). The selection of suitable protecting groups can be readily determined by those skilled in this art. A given reaction can be carried out in a single solvent or a mixture of solvents.

[0102] In the examples, the compounds disclosed herein can be prepared by the following reaction diagrams I to IV. [Reaction Diagram] [I] Synthesis of Epoxide Intermediates [] in: X is H or a cation; LG stands for leaving group, such as halides; n is 1, 2, or 3; p is 1, 2, 3, 4, or 5; Each R1 is independently a halogen, alkyl, or haloalkyl group. [] [Reaction Diagram] [II] General Synthesis of Compounds [] in: n is 1, 2, or 3; p is 1, 2, 3, 4, or 5; m is 1, 2, or 3; q is 1 or 2; and Each R1 is independently a halogroup, alkyl group, or haloalkyl group; R5 is aryl, heteroaryl, alkyl, NH2, NHR A, or NRARB; RA is a C1-6 alkyl, C1-6 alkenyl, or C1-6 alkynyl group, each of which may be substituted with -OH or C1-6 alkoxy groups as appropriate; RB is a C1-6 alkyl, C1-6 alkenyl, or C1-6 alkynyl group, each of which may be substituted with -OH or C1-6 alkoxy groups as appropriate. [Reaction Diagram] [III] General Synthesis of Compounds [] in: n is 1, 2, or 3; p is 1, 2, 3, 4, or 5; q is 1 or 2; and Each R1 is independently a halogroup, alkyl group, or haloalkyl group; R3 is -H, alkyl, alkenyl, or alkynyl; R4 is an alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclic group, each of which may be substituted by one or more R5 groups as appropriate; Alternatively, R3 and R4 may form a heterocyclic group, which may be substituted by one or more R5 groups, depending on the situation. R5 is an aryl, heteroaryl, alkyl, NH2, NHR A or NRARB, or an alkyl group substituted with -NH2; RA can be alkyl, alkenyl, or alkynyl, each of which may be substituted with -OH or alkoxy groups depending on the situation; and RB can be alkyl, alkenyl, or ynyl. [Reaction Diagram] [IV] [:] [make] [Amine deprotecting group] [] [] in: n is 1, 2, or 3; m is 1, 2, 3, 4, or 5; q is 1 or 2; and Each R1 is independently a halogroup, alkyl group, or haloalkyl group. [Treatment methods] []

[0103] In embodiments, the present invention provides a method for treating bacterial infections in an individual in need, comprising administering to the individual one or more pharmaceutically acceptable amounts of one or more compounds of formulas (I), (II), (III), (IV), (V) or their stereoisomers or pharmaceutically acceptable salts. In embodiments, the present invention provides a method for inhibiting bacterial efflux pumps in an individual with a bacterial infection, comprising administering to the individual one or more pharmaceutically acceptable amounts of one or more compounds of formulas (I), (II), (III), (IV), (V) or their stereoisomers or pharmaceutically acceptable salts.

[0104] In this embodiment, the bacterial infection is caused by Gram-negative bacteria. In this embodiment, the Gram-negative bacteria may be intracellular pathogens.

[0105] In the embodiments, the bacterial infection is caused by one or more of the following species: Salmonella spp., Acinetobacter spp., Actinobacter spp., Aeromonas spp., Bacteroides spp., Bodophyllum spp., Brucella spp., Burkholderia spp., Prevotella spp., Porphyromonas spp., Campylobacter spp., Citrobacter spp., Edwardsiella spp., Ekenella spp., Enterobacter spp., Escherichia spp., Franz spp., Haemophilus spp., Helicobacter spp., and Chlorella spp. Caused by the genera *Klebsiella*, *Legionella*, *Moraxella*, *Morganella*, *Neisseria*, *Pasteurella*, *Pseudomonas*, *Proteus*, *Providence*, *Providence*, *Pseudomonas*, *Serratia*, *Shigella*, *Oligotrophomonas*, *Streptococcus*, *Vibrio*, *Yersinia*, *Chlamydia*, *Rickettsia*, *Coxiella*, *Ehrlichia*, or *Bartonia*.

[0106] Acinetobacter baumannii, Acinetobacter haemolyticus, Actinobacillus actinomycetemcomitans, Aeromonas hydrophila, Bacteroides fragilis, Bacteroides theataioatamides theataioatamides theataioatides distasonis, Bacteroides ovatus, Bacteroides vulgatus, Bordetella pertussis, Brucella melitensis, Burkholderia cepacia, Burkholderia pseudomallei, Burkholderia mallei, Prevotella corporis, Prevotella intermedius Intermediate bacteria, *Prevotella endodontalis*, *Porphyromonas asaccharolytica*, *Campylobacter jejuni*, *Campylobacter coli*, *Campylobacter fetus*, *Citrobacter freundii*, *Citrobacter koseri*, *Edwarsiella tarda*, *Eikenella corrodens*, *Enterobacter cloacae*, *Enterobacter aerogenes*, *Enterobacter agglomerans*, *Escherichia coli*, *Francisella tularensis*, *Haemophilus influenzae*, *Haemophilus ducreyi*The following bacteria are listed: *Helicobacter pylori*, *Kingella kingae*, *Klebsiella pella pneumoniae rhinoscleromatis*, *Klebsiella ozaenae*, *Legionella penumophila*, *Moraxella catarrhalis*, *Morganii morganii*, *Neisseria gonorrhoeae*, *Neisseria meningitidis*, *Pasteurella multocida*, *Plesiomonas shigelloides*, *Proteus mirabilis*, *Proteus vulgaris*, *Proteus penneri*, *Proteus myxofaciens*, and *Providencia*. *Providencia rettgeri*, *Providencia alcalifaciens*, *Pseudomonas aeruginosa*, *Pseudomonas fluorescens*, *Salmonella typhi*, *Salmonella enterica*, *Salmonella paratyphi*, *Serratia marcescens*, *Shigella flexneri*, *Shigella boydii*, *Shigella sonnei*, *Shigella dysenteriae*, *Stenotrophomonas maltophilia*, *Streptobacillus moniliformis*, *Vibrio cholerae* cholerae), Vibrio parahaemolyticusThe following bacteria are listed: *Vibrio parahaemolyticus*, *Vibrio vulnificus*, *Vibrio alginolyticus*, *Yersinia enterocolitica*, *Yersinia pestis*, *Yersinia pseudotuberculosis*, *Chlamydophila pneumoniae*, *Chlamydophila trachomatis*, *Rickettsia prowazekii*, *Coxiella burnetii*, *Ehrlichia chaffeensis*, or *Bartonella hensenae*.

[0107] In the embodiments, the bacterial infection is caused by one or more Salmonella spp. In the embodiments, the Salmonella spp. is Enterobacter (a serovar of Salmonella typhimurium). In the embodiments, the bacterial infection is caused by Escherichia coli. In the embodiments, the bacterial infection is caused by Klebsiella pneumoniae. In the embodiments, the bacterial infection is caused by Enterobacter cloacae.

[0108] In the embodiments, the compounds of the present invention may be administered in combination with one or more antibiotics (separately, simultaneously (e.g., as part of the same composition in a combined product), or sequentially). In the embodiments, the antibiotics are macrocyclic lactones, tetracyclines, fluoroquinolones, penicillin, cephalosporins, aminoglycosides, sulfonamides, β-lactams, trimeprine-sulfamethoxazole, chloramphenicol, or lincosamide.

[0109] In the examples, the antibiotics are penicillin G, penicillin V, methicillin, oxacillin, cloxacillin, dicloxacillin, nafcillin, ampicillin, amoxicillin, carbenicillin, ticarcillin, mezlocillin, piperacillin, azlocillin, and temoxicillin. illin, cepalothin, cephapirin, cephradine, cephaloridine, cefazolin, cefamandole, cefuroxime, cephalexin, cefprozil, cefaclor, loracarbef, cefoxitin, cefmatozole, cefotaxime, cefazolin (ceftizoxime), ceftriaxone, cefoperazone, ceftazidime, cefixime, cefpodoxime, ceftibuten, cefdinir, cefpirome, cefepime, BAL5788, BAL9141, imipenem, ertapenem, meropenem, astreonam, clavulanic acid (clavulanate), sulbactam, tazobactam, streptomycin, neomycin, kanamycin, paromycin, gentamicin, tobramycin, amikacin, netilmicin, spectinomycin, sisomicin, dibekalin, isepamicin, tetracycline,Chlortetracycline, demeclocycline, minocycline, oxytetracycline, metacycline, doxycycline, erythromycin, azithromycin, clarithromycin, telithromycin, ABT-773, lincomycin, clindamycin Vancomycin, oritavancin, dalbavancin, teicoplanin, quinupristin, dalfopristin, sulfadiazine, sulfisoxazole, sulfamethoxazole, sulfathalidine, linezolid, nalidixic acid, oxolinic acid (acid), norfloxacin, perfloxacin, enoxacin, ofloxacin, ciprofloxacin, temafloxacin, lomefloxacin, fleroxacin, grepafloxacin, sparfloxacin, trovafloxacin, clinaficin Oxacin), gatifloxacin, moxifloxacin, gemifloxacin, sitafloxacin, metronidazole, daptomycin, garenoxacin, ramoplanin, faropenem, polymyxin, tigecycline, AZD2563, or trimeprilin.

[0110] In the embodiments, the compounds disclosed herein are intended for the treatment of infections caused by Gram-negative bacteria that have developed antibiotic resistance. The terms "resistance" and "bacterial resistance" refer to bacteria that can survive exposure to one or more antibiotics. In the embodiments, the bacteria are resistant to one or more of the following: aminoglycoside antibiotics (e.g., amikacin, gentamicin, kanamycin, neomycin, netilmicin, tobramycin, paromomycin, spectinomycin), ansamycin antibiotics (e.g., rifaximin, streptomycin), carbapenem antibiotics (e.g., ertapenem, doripenem, imipenem / cilastatin, meropenem), cephalosporins. Antibiotics (e.g., cefadroxil, cefaxolin, cefatolin, cefalexin, cefaclor, cefamandole, cefoxitin, cefrozil, cefuroxime, cefisime, cefdinir, cefditoren, cefoperazone, cefotaxime, cefpodoxime, ceftarolin) fosamil, ceftobiprole, glycopeptide antibiotics (e.g., teicoplanin, vancomycin, telavancin), lincomycin antibiotics (e.g., clindamycin, lincomycin), daptomycin, macrocyclic lactone antibiotics (e.g., azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, troleandomycin, telithromycin, spiramycin), aztreonam, furazolidone Nitrofurantoin, oxazolidinone antibiotics (e.g., linezolid, posizolid, radezolid, torezolid), penicillin antibiotics (e.g., amoxicillin, ampicillin, azoxicillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, methicillin, nafcillin, oxacillin, penicillin, piperacillin, temoxicillin, tekcillin), amoxicillin / clavulanic acid, ampicillin / sulbactam, piperacillin / tazobactam, tekcillin / clavulanic acid, quinolone antibacterial agents (e.g.,Ciprofloxacin, enoxacin, gatifloxacin, gemifloxacin, levofloxacin, lomefloxacin, moxifloxacin, nalidixic acid, norfloxacin, ofloxacin, trovafloxacin, grefloxacin, sparfloxacin, temafloxacin), sulfonamide antibiotics (e.g., mafenide, sulfacetamide, sulfadimethoxine, silver sulfadimethoxine, sulfamethizole, sulfamethoxazole, sulfanilimide, sulfasalazine, sulfisoxazole, trimeprine / sulfamethoxazole-TMP-SMX), and tetracycline antibiotics (e.g., demecycline, doxycycline, minocycline, oxytetracycline, tetracycline, tigecycline).

[0111] In the embodiments, when the compounds of the present invention are used in combination with antibiotics, the compounds reduce the MIC (e.g., as measured in Example 2) of the antibiotics by about 1.5 times, about 2 times, about 2.5 times, about 3 times, about 3.5 times, about 4 times, about 4.5 times, about 5 times, about 5.5 times, about 6 times, about 6.5 times, about 7 times, about 7.5 times, about 8 times, about 8.5 times, about 9 times, about 9.5 times, or about 10 times, including all values ​​and ranges therein. In the embodiments, when the compounds of the present invention are used in combination with antibiotics, the dosage of the antibiotics may be reduced (compared to the dosage of the antibiotics administered in the absence of the disclosed compounds) by about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or more, including all values ​​and ranges therein. In the embodiments, the compounds of the present invention allow antibiotics to be effective at the same dosage, at which the antibiotics are ineffective when administered in the absence of one of the disclosed compounds.

[0112] In the embodiments, the present invention provides a method for increasing the sensitivity of bacteria to antibiotic treatment by administering the compound of the present invention and one or more antibiotics.

[0113] In embodiments, the method includes administering to an individual in need one or more pharmaceutically acceptable amounts of one or more of formulas (I), (II), (III), (IV), (V) or their stereoisomers or pharmaceutically acceptable salts and one or more antibiotics. In embodiments, the method increases the susceptibility of Gram-negative bacteria to antibiotics. In embodiments, the compounds of the present invention can suscepte antibiotic-resistant strains of bacteria to antibiotics they were originally resistant to. In embodiments, the antibiotic is a macrocyclic lactone, tetracycline, fluoroquinolone, penicillin, cephalosporin, aminoglycoside, sulfonamide, β-lactam, trimeprine-sulfamethoxazole, chloramphenicol, or lincosamide.

[0114] In the embodiments, when the compounds of the present invention are used in combination with antibiotics, the compounds increase the sensitivity of the antibiotics by reducing the MIC of the antibiotics by about 1.1 times, about 1.2 times, about 1.3 times, about 1.4 times, about 1.5 times, about 2 times, about 2.5 times, about 3 times, about 3.5 times, about 4 times, about 4.5 times, about 5 times, about 5.5 times, about 6 times, about 6.5 times, about 7 times, about 7.5 times, about 8 times, about 8.5 times, about 9 times, about 9.5 times, or about 10 times (inclusive of all values ​​and ranges). In the embodiments, when the compounds of the present invention are used in combination with antibiotics, these compounds increase antibiotic sensitivity by reducing the dosage and / or IC 50 (compared to the dosage of antibiotics administered in the absence of the disclosed compounds) by about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90% or more (inclusive of all values ​​and ranges therein). In the embodiments, the compounds of the present invention allow antibiotics to be effective at the same dosage at which they would be ineffective when administered in the absence of one of the disclosed compounds.

[0115] In embodiments, the present invention also provides a method for reversing or reducing antibiotic resistance in antibiotic-resistant Gram-negative bacteria. In embodiments, the method includes administering to an individual in need one or more pharmaceutically acceptable amounts of one or more antibiotics of formulas (I), (II), (III), (IV), (V) or their stereoisomers or pharmaceutically acceptable salts. In embodiments, the antibiotic is a macrocyclic lactone, tetracycline, fluoroquinolone, penicillin, cephalosporin, aminoglycoside, sulfonamide, β-lactam, trimeprine-sulfamethoxazole, chloramphenicol, or lincosamide.

[0116] In the embodiments, the reversal or reduction of antibiotic resistance is determined by assessing the decrease in the growth rate of antibiotic-resistant strains in the presence of the compound and one or more antibiotics. In the embodiments, administration of the compound of the present invention in combination with antibiotics reverses or reduces antibiotic resistance by reducing bacterial pathogenicity, inhibiting or killing antibiotic-resistant bacteria, preventing biofilm formation, preventing septic shock, treating sepsis, and / or increasing bacterial susceptibility to previously exhibited resistance to antibiotics. In the embodiments, when the compound of the present invention is used in combination with antibiotics, the pathogenicity or growth rate of antibiotic-resistant bacteria may be reduced (compared to the dose of antibiotic administered in the absence of the disclosed compound) by about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or more (inclusive of all values ​​and ranges therein). In the embodiments, when the compounds of the present invention are used in combination with antibiotics, the compounds reverse or reduce antibiotic resistance by reducing the MIC of the antibiotic by about 1.1 times, about 1.2 times, about 1.3 times, about 1.4 times, about 1.5 times, about 2 times, about 2.5 times, about 3 times, about 3.5 times, about 4 times, about 4.5 times, about 5 times, about 5.5 times, about 6 times, about 6.5 times, about 7 times, about 7.5 times, about 8 times, about 8.5 times, about 9 times, about 9.5 times, or about 10 times (inclusive of all values ​​and ranges). [Pharmaceutical Compositions] []

[0117] In some embodiments of the present invention, the pharmaceutical composition comprises a therapeutically effective amount of one or more of the formulas (I), (II), (III), (IV) or (V) compounds or stereoisomers thereof or pharmaceutically acceptable salts.

[0118] In the embodiments, pharmaceutical compositions comprising one or more compounds disclosed herein, or their stereoisomers or pharmaceutically acceptable salts, and pharmaceutically acceptable excipients or adjuvants are provided. Pharmaceutically acceptable excipients and adjuvants are added to the composition or formulation for various purposes. In the embodiments, the pharmaceutical composition comprises pharmaceutically acceptable carriers, binders, and / or diluents. In the embodiments, the pharmaceutical composition may contain additional materials suitable for the physical formulation of various dosage forms of the compositions of the present invention, such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickeners, stabilizers, lubricants, wetting agents, emulsifiers, salts affecting osmotic pressure, buffers, colorants, flavorings, and / or aromatic substances.

[0119] In some embodiments, the pharmaceutical compositions of the present invention may additionally contain other auxiliary components conventionally found in pharmaceutical compositions and established at their respective levels of use in the relevant art. Thus, for example, such pharmaceutical compositions may contain additional compatible pharmaceutically active substances, such as antipruritics, astringents, local anesthetics, or anti-inflammatory agents.

[0120] The compounds of this invention can be formulated into formulations containing pharmaceutically acceptable carriers, adjuvants, and mediators for administration by various methods, including oral and enteral administration. As used herein, the term enteral administration includes subcutaneous, intravenous, intramuscular, and intra-arterial injection using various infusion techniques. As used herein, intra-arterial and intravenous injection include administration via catheter.

[0121] The compounds disclosed herein can be formulated according to conventional procedures applicable to the desired route of administration. Therefore, the compounds disclosed herein can be in the form of suspensions, solutions, or emulsions contained in oily or aqueous media, and may contain suspending agents, stabilizers, and / or dispersants. The compounds disclosed herein can also be formulated into injection formulations.

[0122] In some embodiments, the pharmaceutical compositions of the present invention are prepared using known techniques, including (but not limited to) mixing, dissolving, granulation, sugar-coated pill making, water milling, emulsification, encapsulation, trapping, or tablet making processes.

[0123] In some embodiments, the pharmaceutical composition may be a solid, powder, liquid, or gel. In some embodiments, the pharmaceutical agent is a solid (e.g., powder, tablet, capsule, granules, and / or aggregate). In some of these embodiments, the solid pharmaceutical composition comprises one or more excipients known in the art, including (but not limited to) starch, sugar, diluent, granulating agent, lubricant, binder, and disintegrant.

[0124] Solid carriers applicable to this application include (but are not limited to) sugars and sugar alcohols (e.g., lactose, glucose, mannitol, and the like), starch, methylcellulose, magnesium stearate, dicalcium phosphate, calcium phosphate, talc, sugar, dextran, starch, gelatin, cellulose, and polyvinylpyrrolidone. Solid carriers may further comprise one or more substances that act as flavoring agents, lubricants, solubilizers, suspending agents, fillers, flow aids, compression aids, binders, or tablet disintegrants. Tablets may be prepared by compression or molding, as appropriate, with one or more auxiliary ingredients. Compressed tablets may be prepared by compressing an active ingredient in a free-flowing form (such as powder or granules) mixed with, as appropriate, a binder (e.g., povidone, gelatin, hydroxypropyl methylcellulose), lubricant, inert diluent, preservative, and / or disintegrant (e.g., sodium starch glycolate, crospovidone, crospovidone carboxymethyl cellulose sodium) in a suitable machine. Molding tablets can be prepared by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0125] In the embodiments, the pharmaceutical composition is formulated into a liquid. Liquid pharmaceutical compositions suitable for use in this invention include solutions, suspensions, emulsions, syrups, elixirs, and pressurized compounds. The active ingredient can be dissolved or suspended in a pharmaceutically acceptable liquid carrier, such as water, an organic solvent, a mixture of both, or a pharmaceutically acceptable oil or fat. In the embodiments, the liquid solution can be an aqueous or non-aqueous solution.

[0126] Examples of non-aqueous carriers include (but are not limited to) propylene glycol, polyethylene glycol, vegetable oils (such as olive oil), and injectable organic esters.

[0127] The aqueous carriers applicable to this application include (but are not limited to) water, ethanol, alcohol / aqueous solutions, glycerol, emulsions or suspensions (including saline and buffer media).

[0128] Non-enteral carriers applicable to this application include (but are not limited to) sodium chloride solution, Ringer's dextran, dextran and sodium chloride, lactated Ringer's, and fixed oil. Intravenous carriers include fluids and nutritional supplements, electrolyte supplements, such as those based on Ringer's dextran, and the like. Preservatives and other additives may also be present, such as, for example, antibacterial agents, antioxidants, chelating agents, inert gases, and the like. Liquid carriers may contain other suitable pharmaceutical additives, such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickeners, coloring agents, viscosity modifiers, stabilizers, or osmotic pressure modifiers.

[0129] In the embodiments, the amount of the compound disclosed herein or its stereoisomers or pharmaceutically acceptable salts may be administered from about 0.001 mg / kg to about 100 mg / kg body weight (e.g., about 0.01 mg / kg to about 10 mg / kg or about 0.1 mg / kg to about 5 mg / kg). In the embodiments, the amount of the compound disclosed herein or its stereoisomers or pharmaceutically acceptable salts may be administered from about 0.1 mg to about 1,000 mg (e.g., about 0.1 mg to about 500 mg / kg or about 0.1 mg / kg to about 100 mg / kg). [Example] []

[0130] The following experiments were performed using the SAFIRE analysis described in U.S. Patent Publication No. 2020 / 0022961, the entire contents of which are incorporated herein by reference for all purposes. SAFIRE (screening for intracellular anti-infective agents of Enterobacteriaceae using fluorescence microscopy) uses cell cultures as a substitute to identify compounds that prevent intracellular replication of human pathogens, model Gram-negative bacteria. This method identifies compounds that prevent bacterial growth in culture media and at concentrations... [<] Compounds that reduce the load of Gram-negative bacteria inside macrophages at 10 µM, but have no effect on bacterial growth in standard Mueller Hinton (MHB) medium at 100 µM. Therefore, SAFIRE enables the discovery of compounds that are effective against Gram-negative bacteria in host cells and all animals, regardless of whether they are antibacterial in culture medium. [Example] [1.] [Inhibition Curve] []

[0131] [Bacterial strain] []

[0132] Wild-type Enterobacter spp. (a serovar of Salmonella typhimurium) strain SL1344 was initially isolated from the blood of infected calves. For screening and validation in macrophages, SL1344 sifB::gfp was grown overnight to saturation in Luria-Bertani medium (LB) containing 30 μg / ml streptomycin and 30 μg / ml kanamycin, diluted to 0.001 OD, and aliquots were frozen in 20% glycerol at -80°C. Before infection, aliquots were grown at 37°C in 5 mL LB culture containing 30 μg / ml streptomycin and 30 μg / ml kanamycin for 18 hours with aeration. The bacterial strain was routinely grown in LB with antibiotics: 30 μg / ml streptomycin, 30 μg / ml kanamycin, 50 μg / ml ampicillin, 10 μg / ml tetracycline, and / or 1.15 μg / ml meropenem. The acrAB::kan and macAB::kan strains were constructed using known methods. The enteric Salmonella subgenus and mouse typhoid bacillus serovars S10801 and NR-22067 were multidrug-resistant isolates from septic calves. These strains, and others as specified, were obtained through BEI resources from NIAID and NIH. []

[0133] [Cell Culture] [thing] []

[0134] RAW 264.7 murine macrophage cells and the HeLa human epithelial cell line were obtained from the American Type Tissue Collection. The BMDM line was isolated as previously described. Briefly, bone marrow was extracted from the femur of 1- to 4-month-old internally fed 129SvEvTac mice (Taconic Laboratory). Monocytes were isolated using Histopaque-1083 (Sigma), washed, and directly seeded into analytical plates at 1 x 10⁵ cells / ml in complete medium supplemented with 35% conditioned medium from MCSF-expressing 3T3 cells. The medium was replaced after three days. After one week, the medium was replaced with 100 μL of fresh medium and the cells were infected as described below. All three cell types were grown in DMEM high glucose (Sigma) supplemented with 10% fetal bovine serum, 2 mM L-glutamylamine, 1 mM sodium pyruvate, 10 mM HEPES, and 50 μM β-mercaptoethanol. Cells were maintained at 37°C under a humidified atmosphere of 5% CO2. For screening, frozen aliquots of RAW 264.7 were thawed and allowed to expand for three days prior to inoculation; other experiments were performed using cultures between passage 4 and passage 20. []

[0135] [For use] [SAFIRE] [Bacterial infection and] [CFU] [Platelet inoculation] []

[0136] SAFIRE-RAW264.7 macrophages (7 x 10³ cells in 40 μL or 5 x 10⁴ cells in 100 μL) were seeded individually in 384- or 96-well Brooks Automation black-walled glass plates. 24 hours post-inoculation, bacteria in 20 or 50 μL PBS were added to a final concentration of 1 x 10⁷ CFU / mL, which resulted in approximately 70% macrophage infection and minimal macrophage cytotoxicity 18 hours post-inoculation. A sifB::gfp bacterial reporter line was used to minimize the green signal from extracellular bacteria. 45 minutes after bacterial addition, 20 or 50 μL of gentamicin was added to a final concentration of 40 μg / mL, which did not affect intracellular infection but inhibited extracellular bacterial replication. 2 hours post-inoculation, 200 or 500 nL of the compound was added using a pin tool (CyBio) to produce a final concentration of 25 μM. Each analytical plate included rifampicin and DMSO controls. In some experiments, the culture medium was removed and replaced with fresh medium containing 40 μg / mL gentamicin and an increased concentration of the compound of the present invention. At 17.5 hours post-infection, PBS containing MitoTracker Red CMXRos (Life Technologies) was added to the 384- or 96-well plates to a final concentration of 300 nM or 100 nM, respectively. Thirty minutes later, 16% paraformaldehyde was added to 1 ± 2% to the final concentration and incubated at room temperature for 15 minutes. The wells were washed twice with PBS and stained with 1 μM DAPI for 20 minutes; the wells were washed twice and stored in PBS containing 90% glycerol until imaging. The Z'-factors of the screening platform for the 96-well and 384-well plates were 0.59 and 0.48, respectively, within the range disclosed for complex cell-based screening. []

[0137] HeLa cells were infected with Salmonella typhimurium as described above, except that 1 x 10⁴ cells were seeded and the cells were infected with Salmonella typhimurium that constitutively expressed GFP from the rpsM locus, because sifB::gfp does not express well in HeLa cells. Furthermore, after adding the bacteria, the plate was rotated at 500 x g for 5 minutes to increase infection. []

[0138] CFU infection was performed as described above, except that cells were seeded in 96-well Greiner plates coated with tissue culture. Eighteen hours post-infection, the wells were washed three times with PBS, dissolved in 30 μL of 0.1% Triton X-100, diluted, and plated to determine CFU levels.

[0139] The IC50 values ​​of the compounds of this invention were measured using SAFIRE across at least eight concentrations ranging from 50 to 0.001 µM and are provided in Table A below.

[0140] Furthermore, toxicity was measured according to the following procedures: 1) macrophage morphology analysis; and 2) cell counting using MATLAB (cells that rose and floated after treatment were not within the appropriate Z-plane for counting and were therefore considered dead). A compound was considered non-toxic if 70% of the cell lines adhered (as measured by comparing cell numbers before and after treatment). Compounds passing #1 and #2 above were considered to have toxicity >50 μM. Compounds failing #1 or #2 were considered to have toxicity <25 μM. Data are shown in Table A below. [surface] [A] [] [Compound Number] [structure] [IC, 50 , (µM) ] [toxicity] [(µM)] 1 <1> 50 2 < 1 > 50 3 < 1 > 50 4 < 1 < 25 5 < 1 > 50 6 < 1 > 50 7 < 1 <50 but> 25 8 < 1 > 50 9 < 1 > 50 10 < 1 <50 but> 25 11 < 1 > 50 12 < 1 > 50 13 < 1 > 50 14 > 1 > 50 15 < 1 > 50 16 < 1 > 50 17 < 1 < 25 18 < 1 > 50 19 < 1 <50 but> 25 [Example] [2.] [Culture medium activity profile] []

[0141] To evaluate the ability of the compounds of this invention to susceptible bacteria to antibiotics, the compounds of this invention were combined with known antibiotics doxycycline, ciprofloxacin, and chloramphenicol in several bacterial cell lines (including carbapenem-resistant Enterobacteriaceae), and the minimum inhibitory concentration (MIC) was measured. The MIC of the antibiotic alone (i.e., in the absence of the disclosed compound) is provided in the first row and marked "None". The MICs of the antibiotics were re-evaluated in each cell line by adding 50 µg / mL of comparative compounds EPI 35 and PaβN, which have been reported to inhibit bacterial efflux pumps. These results are presented in Tables B through D. When used at a concentration of 50 µg / mL, the compounds of this invention reduced the MICs of these antibiotics. At this concentration (50 µg / mL), the compounds of this invention lacked intrinsic antibacterial activity, but a significant improvement in efficacy was still observed. Therefore, the compounds of this invention can be used to enhance the activity of antibiotics that have lost their efficacy due to efflux. [surface] [B] [Selected compound] [Doxycycline] [MIC (µg / mL)] 50 µg / mL contained in MHB Salmonella S10801 E. coli BAA-2340 Klebsiella pneumoniae BAA-2342 Klebsiella pneumoniae BAA-1705 Enterobacter cloacae BAA-2341 none 32 8 8 4 8 AND 35 8 8 8 8 8 1 8 [<]1 2 [<]1 [<]1 3 8 [<]1 2 [<]1 [<]1 4 8 [<]1 [<]1 [<]1 [<]1 7 8 [<]1 [<]1 [<]1 [<]1 10 8 [<]1 [<]1 [<]1 [<]1 16 8 [<]1 2 [<]1 [<]1 17 8 2 4 [<]1 2 18 4 [<]1 [<]1 [<]1 [<]1 19 8 [<]1 [<]1 [<]1 [<]1 PAβN (25 µg / mL) 8 [<]1 [<]1 [<]1 [<]1 [surface] [C] [Selected Compounds] [PMB MIC (µg / mL)] [Ciprofloxacin] [MIC (µg / mL)] 50 µg / mL contained in MHB Enterobacteriaceae MB315 Escherichia coli BAA-2340 Klebsiella pneumoniae BAA-2342 Klebsiella pneumoniae BAA-1705 Enterobacter cloacae BAA-2341 none 32 32 32 >64 32 EPI 35 1 32 32 64 32 1 32 8 8 32 16 3 32 16 16 32 16 4 16 8 8 16 8 7 32 8 8 32 16 10 32 16 16 32 16 16 32 16 16 32 16 17 16 16 16 32 16 18 1 8 8 16 8 19 32 32 32 64 16 PAβN (25 µg / mL) 32 16 16 64 16 [surface] [D] [Selected compound] Chloramphenicol [MIC (µg / mL)] 50 µg / mL contained in MHB E. coli BAA-2340 Klebsiella pneumoniae BAA-2342 Enterobacter cloacae BAA-2341 Acinetobacter baumannii MB306 Acinetobacter baumannii MB309 Pseudomonas aeruginosa focus MB0477 none 64 64 16 >64 >64 64 EPI 35 64 64 16 64 64 >64 1 16 16 2 64 64 64 3 16 16 2 64 64 64 4 8 8 2 64 64 64 7 8 8 2 32 64 64 10 8 8 2 32 64 4 16 16 16 2 32 64 64 17 16 16 4 32 64 64 18 8 8 2 32 32 >64 19 16 16 2 64 64 16 PAβN (25 µg / mL) 8 8 2 32 64 1 [Example] [3.] [Methods for preparing compounds] []

[0142] The compounds disclosed herein can be prepared by the general synthetic reaction diagrams I to IV shown below and / or by any other suitable method. These compounds can be characterized by any suitable method known in this art, such as NMR, UV, HPLC, LC-MS, and TLC. [Reaction Diagram] [I] Synthesis of Epoxide Intermediates [] in: X is H or a cation; LG stands for leaving group, such as halides; n is 1, 2, or 3; p is 1, 2, 3, 4, or 5; and Each R1 is independently a halogen, alkyl, or haloalkyl group.

[0143] A stirred mixture of 1.0 g of 3,4-fluorophenol, 1.5 equivalents of epichlorohydrin, and 2.0 equivalents of Cs₂CO₃ was heated to 80 °C overnight. The reaction was stopped with water, and the precipitate was purified by normal-phase column chromatography using ethyl acetate and hexane, yielding good results. The product was used in the next step. The product was characterized by LC-MS, and the desired mass was observed. [Reaction Diagram] [II] General Synthesis of Compounds [] in: n is 1, 2, or 3; p is 1, 2, 3, 4, or 5; m is 1, 2, or 3; q is 1 or 2; and Each R1 is independently a halogroup, alkyl group, or haloalkyl group; R5 is aryl, heteroaryl, alkyl, NH2, NHR A, or NRARB; RA is a C1-6 alkyl, C1-6 alkenyl, or C1-6 alkynyl group, each of which may be substituted with -OH or C1-6 alkoxy groups as appropriate; and RB is a C1-6 alkyl, C1-6 alkenyl, or C1-6 alkynyl group, each of which may be substituted with -OH or C1-6 alkoxy groups as appropriate.

[0144] Two starting materials (0.16 mmol) were mixed in 1 mL of DI water at a 1:1 mol ratio and heated to 140 °C for 10 minutes using a microwave oven. The resulting product was purified by reverse-phase column chromatography. The product was characterized by LC-MS. The desired mass was observed, as shown below. [surface] As shown in [E]. [] [Reaction Diagram] [III] General Synthesis of Compounds [] in: n is 1, 2, or 3; p is 1, 2, 3, 4, or 5; q is 1 or 2; and Each R1 is independently a halogroup, alkyl group, or haloalkyl group; R3 is -H, alkyl, alkenyl, or alkynyl; R4 is an alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclic group, each of which may be substituted by one or more R5 groups as appropriate; Alternatively, R3 and R4 may form a heterocyclic group, which may be substituted by one or more R5 groups, depending on the situation. R5 is an aryl, heteroaryl, alkyl, NH2, NHR A or NRARB, or an alkyl group substituted with -NH2; RA can be alkyl, alkenyl, or alkynyl, each of which may be substituted with -OH or alkoxy groups depending on the situation; and RB can be alkyl, alkenyl, or ynyl.

[0145] Two starting materials (0.16 mmol) were mixed in 1 mL of DI water at a 1:1 mol ratio and heated to 140 °C for 10 minutes using a microwave oven. The resulting product was purified by reverse-phase column chromatography. The product was characterized by LC-MS. The desired mass was observed, as shown below. [surface] As shown in [E]. [] [Reaction Diagram] [IV] [:] [make] [Amine deprotecting group] [] [] in: n is 1, 2, or 3; m is 1, 2, 3, 4, or 5; q is 1 or 2; and Each R1 is independently a halogroup, alkyl group, or haloalkyl group.

[0146] The starting material (1.11 mmol) was dissolved in 3 mL of 4N HCl in dioxane and stirred overnight. The solvent was removed by evaporation, and the final product was precipitated from diethyl ether, filtered, and dried to obtain the product as a diHCl salt. The product was characterized by LC-MS. The desired mass was observed, as shown below. [surface] As shown in [E]. [surface] [E.] [Measuring the mass and synthesis methods of compounds] [] [serial number] [structure] [MW (g / mol)] [Measurement value] [(g / mol)] [Synthetic Reaction Diagram] 1 293 294 I、II、IV 2 293 294 I、II、IV 3 334 335 I、II 4 334 335 I、II 5 336 337 I、II 6 336 337 I、II 7 336 337 I、II 8 336 337 I、II 9 333 334 I、II、IV 10 377 378 I、II 11 350 351 I、II 12 394 395 I、II 13 394 395 I、II 14 394 395 I、II 15 336 337 I、II 16 334 335 I、II 17 356 357 I、II 18 331 332 I、III、IV 19 319 320 I、III、IV

Claims

1. A compound having the structure of formula (IV) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, , (IV) wherein: n is 1 or 2; q is 1; each R1 is Cl or fluoroalkyl; R5 is alkyl-NH2, -NH2, -NHRA or NRARB; RA is a C1-6 alkyl group substituted with C1-6 alkoxy as appropriate; and RB is a C1-6 alkyl group.

2. The compound of claim 1 or its stereoisomer or pharmaceutically acceptable salt thereof, wherein R5 is an alkyl-NH2 or -NH2.

3. The compound of claim 1 or its stereoisomer or pharmaceutically acceptable salt, wherein the compound has the following structures: , , , , or.

4. A compound having the structure of formula (V) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein: n is 1 or 2; q is 1 or 2; each R1 is Cl or fluoroalkyl; R5 is NRARB or imidazolyl; RA is a C1-6 alkyl group substituted with C1-6 alkoxy; and RB is a C1-6 alkyl group.

5. A compound of claim 4 or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein the compound has the following structures: , or.

6. A compound having one of the following structures, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, , , , , , or.

7. A compound having the following structure or a stereoisomer thereof or a pharmaceutically acceptable salt thereof.

8. A compound having the following structure or a stereoisomer thereof or a pharmaceutically acceptable salt thereof.

9. A pharmaceutical composition comprising a therapeutically effective amount of any one of claims 1 to 8 of the compound or its stereoisomer or pharmaceutically acceptable salt and one or more pharmaceutically acceptable excipients.

10. A pharmaceutical composition comprising a therapeutically effective amount of a compound having the following structure or a stereoisomer thereof or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients.

11. Use of a compound or stereoisomer of any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating bacterial infections.

12. As requested in claim 11, wherein the bacterial infection is caused by an intracellular pathogen.

13. As requested in claim 11, wherein the bacterial infection is caused by Gram-negative bacteria.

14. As claimed in claim 13, wherein the bacterial infection is caused by one or more of the following species: Salmonella sp., Acinetobacter sp., Actinobacillus sp., Aeromonas sp., Bacteroide sp., Bordetella sp., Brucella sp., Burkholderia sp., Prevotella sp., Porphyromonas sp., Campylobacter sp., Citrobacter sp., Edwarsiella sp., Eikenella sp., Enterobacter sp., Escherichia sp., and Francisella sp. sp.), Haemophilus sp., Helicobacter sp., Kingella sp., Klebsiella sp., Legionella sp., Moraxella sp., Morganella sp., Neisseria sp., Pasteurella sp., Plesiomonas sp., Proteus sp., Providencia sp., Pseudomonas sp., Serratia sp., Shigella sp., Stenotrophomonas sp., Streptobacillus sp., Vibrio sp., Yersinia sp. Caused by the genera *Chlamydophila*, *Ricketsia*, *Coxiella*, *Ehrlichia*, or *Bartonella*.

15. As claimed in claim 14, wherein the bacterial infection is caused by one or more Salmonella species.

16. As claimed in claim 15, wherein the Salmonella spp. is enterica (a serovar of Salmonella typhimurium in mice).

17. As claimed in claim 11, wherein the bacterial infection is caused by Escherichia coli, Klebsiella pneumoniae, or Enterobacter cloacae.

18. As claimed in claim 11, wherein the compound is: or a stereoisomer thereof or a pharmaceutically acceptable salt thereof.

19. As claimed in claim 11, wherein the compound is...

20. Use of a compound having the following structure or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating bacterial infections.

21. As claimed in claim 11, wherein the bacterial infection is resistant to one or more antibiotics.

22. As claimed in claim 11, wherein the agent is used in combination with one or more antibiotics.

23. As claimed in claim 22, wherein the antibiotic is a macrolide, tetracycline, fluoroquinolone, penicillin, cephalosporin, aminoglycoside, sulfonamide, β-lactam, trimethoprim-sulfamethoxazole, chloramphenicol, or lincosamide.

24. Use of a compound or stereoisomer of any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof and one or more antibiotics in the manufacture of an agent for treating bacterial infections.

25. As requested in claim 24, wherein the bacterial infection is caused by an intracellular pathogen.

26. As requested in claim 24, wherein the bacterial infection is caused by a Gram-negative bacterium.

27. As claimed in claim 26, wherein the bacterial infection is caused by one or more of the following species: Salmonella spp., Acinetobacter spp., Actinobacter spp., Aeromonas spp., Bacteroides spp., Bodophyllum spp., Brucella spp., Burkholderia spp., Prevotella spp., Porphyromonas spp., Campylobacter spp., Citrobacter spp., Edwardsiella spp., Ekenella spp., Enterobacter spp., Escherichia spp., Franz spp., Haemophilus spp., and Helicobacter spp. Caused by the genera *Aureobacterium*, *Klebsiella*, *Lactobacillus*, *Moraxella*, *Morganella*, *Neisseria*, *Pasteurella*, *Pseudomonas*, *Proteus*, *Providence*, *Providence*, *Pseudomonas*, *Serratia*, *Shigella*, *Oligotrophomonas*, *Streptococcus*, *Vibrio*, *Yersinia*, *Chlamydia*, *Rickettsia*, *Coxiella*, *Ehrlich*, or *Bartonia*.

28. As claimed in claim 27, wherein the bacterial infection is caused by one or more Salmonella species.

29. As requested in claim 28, wherein the Salmonella genus is Enterobacter (a serovar of Salmonella typhimurium in mice).

30. As claimed in claim 24, wherein the bacterial infection is caused by Escherichia coli, Klebsiella pneumoniae or Enterobacter cloacae.

31. The use as claimed in claim 24, wherein the antibiotic is a macrocyclic lactone, tetracycline, fluoroquinolone, penicillin, cephalosporin, aminoglycoside, sulfonamide, β-lactam, trimeprine-sulfamethoxazole, chloramphenicol, or lincosamide.

32. Use of a compound or stereoisomer of any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for inhibiting bacterial efflux pumps in an individual with a bacterial infection.

33. As claimed in claim 32, wherein the bacterial infection is caused by an intracellular pathogen.

34. As claimed in claim 33, wherein the bacterial infection is caused by Gram-negative bacteria.

35. As claimed in claim 34, wherein the bacterial infection is caused by one or more of the following species: Salmonella spp., Acinetobacter spp., Actinobacter spp., Aeromonas spp., Bacteroides spp., Bodophyllum spp., Brucella spp., Burkholderia spp., Prevotella spp., Porphyromonas spp., Campylobacter spp., Citrobacter spp., Edwardsiella spp., Ekenella spp., Enterobacter spp., Escherichia spp., Franz spp., Haemophilus spp., and Helicobacter spp. Caused by the genera *Aureobacterium*, *Klebsiella*, *Lactobacillus*, *Moraxella*, *Morganella*, *Neisseria*, *Pasteurella*, *Pseudomonas*, *Proteus*, *Providence*, *Providence*, *Pseudomonas*, *Serratia*, *Shigella*, *Oligotrophomonas*, *Streptococcus*, *Vibrio*, *Yersinia*, *Chlamydia*, *Rickettsia*, *Coxiella*, *Ehrlich*, or *Bartonia*.

36. As claimed in claim 35, wherein the bacterial infection is caused by one or more Salmonella species.

37. As requested in claim 36, wherein the Salmonella genus is Enterobacter (a serovar of Salmonella typhimurium in mice).

38. The use as claimed in claim 32, wherein the compound is a stereoisomer of itself or a pharmaceutically acceptable salt thereof.

39. The use as claimed in claim 32, wherein the compound is a stereoisomer of itself or a pharmaceutically acceptable salt thereof.

40. As claimed in claim 32, wherein the bacterial infection is resistant to one or more antibiotics.

41. The use as claimed in claim 32, wherein the agent is used in combination with one or more antibiotics.

42. The use as claimed in claim 41, wherein the antibiotic is a macrocyclic lactone, tetracycline, fluoroquinolone, penicillin, cephalosporin, aminoglycoside, sulfonamide, β-lactam, trimeprine-sulfamethoxazole, chloramphenicol, or lincosamide.

43. Use of a compound having the following structure or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for inhibiting bacterial efflux pumps in an individual with a bacterial infection.

44. A method for increasing the sensitivity of Gram-negative bacteria to antibiotics in vitro, comprising combining administration of a compound or stereoisomer of any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof with an antibiotic.

45. A method for reversing or reducing antibiotic resistance in living Gram-negative bacteria, comprising combining administration of a compound or stereoisomer thereof or a pharmaceutically acceptable salt thereof, as claimed in any one of claims 1 to 8, with an antibiotic.

46. ​​The method of claim 44, wherein the antibiotic is a macrocyclic lactone, tetracycline, fluoroquinolone, penicillin, cephalosporin, aminoglycoside, sulfonamide, β-lactam, trimeprine-sulfamethoxazole, chloramphenicol, or lincosamide.

47. The method of claim 45, wherein the antibiotic is a macrocyclic lactone, tetracycline, fluoroquinolone, penicillin, cephalosporin, aminoglycoside, sulfonamide, β-lactam, trimeprine-sulfamethoxazole, chloramphenicol, or lincosamide.