Use of compounds in mycobacterium tuberculosis infection

Abstract

The present application belongs to the technical field of medical use, and particularly relates to the use of compounds in mycobacterium tuberculosis infection. The present application provides the use of a compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating mycobacterium tuberculosis infection:

Description

[0001] This application claims priority to the prior application filed on November 29, 2024, with China National Intellectual Property Administration, patent application number 202411741143.8, entitled "Use of a compound in Mycobacterium tuberculosis infection"; the entire contents of the prior application are incorporated herein by reference. Technical Field

[0002] This invention belongs to the field of pharmaceutical application technology, specifically relating to the use of compounds in Mycobacterium tuberculosis infection. Background Technology

[0003] Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (Mtb) and remains one of the major global health threats in the 21st century. TB is primarily transmitted through the air; when a TB patient coughs, sneezes, or talks, they release droplets containing Mycobacterium tuberculosis. When healthy individuals inhale these droplets, the Mycobacterium tuberculosis can multiply in the lungs or other organs, leading to infection. The clinical manifestations of TB are diverse, ranging from asymptomatic to severe systemic disease. Because of the long incubation period, patients may not show any symptoms for a considerable time, complicating early diagnosis and treatment.

[0004] According to the World Health Organization (WHO) 2024 Tuberculosis Report, an estimated 8.2 million people were newly diagnosed with tuberculosis globally in 2023, and approximately 1.25 million died from the disease, making it one of the leading causes of death worldwide. The increasing prevalence of multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) has become a major obstacle to global tuberculosis control efforts (WHO 2024, Ormerod 2005, Raviglione and Smith 2007). The emergence of these drug-resistant tuberculosis diseases is mainly due to improper drug use, treatment interruptions or incomplete courses, and increased drug tolerance in tuberculosis patients.

[0005] Therefore, there is an urgent need to develop new drugs for Mycobacterium tuberculosis infection in order to improve treatment outcomes and improve treatment options for drug-resistant tuberculosis. Summary of the Invention

[0006] This invention provides the use of a compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention and / or treatment of Mycobacterium tuberculosis infection:

[0007] Wherein, R1 is selected from those without substitution or by one or more R a Replacement C 3-12 cycloalkyl, wherein each R a They may be the same or different, and are independently selected from hydroxyl, halogen, cyano, and C groups. 1-12 Alkyl, C 1-12 Alkyloxy; R2 and R7 may be the same or different, and are independently selected from those without substitution or by one or more R... b Replacement C 1-12 Alkyl groups, wherein each R b They may be the same or different, and are independently selected from hydroxyl, halogen, cyano, and C groups. 1-12 Alkyl, C 1-12 Alkyloxy, C 3-12 cycloalkyl; R3, R4, R5, and R6 may be the same or different, and are independently selected from H, hydroxyl, halogen, cyano, and C. 1-12 Alkyl, C 1-12 Alkyloxy, C 3-12 cycloalkyl; R8 and R9 may be the same or different, and are independently selected from H, unsubstituted, or by one or more R... c The following groups are substituted: C 1-12 Alkyl, C 3-12 cycloalkyl, wherein each R c They may be the same or different, and are independently selected from hydroxyl, halogen, cyano, and C groups. 1-12 Alkyl, C 1-12 Alkyloxy, C 3-8 cycloalkyl; R 10 Selected from H or C 1-12 alkyl.

[0008] According to an embodiment of the invention, R1 is selected from those without substitution or by one or more R... a Replacement C 3-8 Cycloalkyl, preferably unsubstituted or with one or more R a Replacement C 3-6 Cycloalkyl, such as unsubstituted or substituted with one or more R a The following groups may be substituted: cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0009] Preferably, each R a They may be the same or different, and are independently selected from hydroxyl, halogen, cyano, and C groups. 1-6 Alkyl, C 1-6 Alkyloxy group.

[0010] According to an embodiment of the invention, R2 and R7 may be the same or different, and are independently selected from those without substitution or by one or more R...b Replacement C 1-6 Alkyl groups, such as unsubstituted or substituted with one or more R groups b The following groups are substituted: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl.

[0011] Preferably, each R b They may be the same or different, and are independently selected from hydroxyl, halogen, cyano, and C groups. 1-6 Alkyl, C 1-6 Alkyloxy, C 3-6 Cycloalkyl.

[0012] According to embodiments of the present invention, R3, R4, R5, and R6 may be the same or different, and are independently selected from H, hydroxyl, halogen, cyano, and C. 1-6 Alkyl, C 1-6 Alkyloxy, C 3-6 Cycloalkyl, preferably H.

[0013] According to embodiments of the present invention, R8 and R9 may be the same or different, and are independently selected from H, unsubstituted, or by one or more R... c The following groups are substituted: C 1-6 Alkyl, C 3-6 Cycloalkyl groups, preferably selected from H, unsubstituted, or substituted with one or more R groups. c Replacement C 1-6 Alkyl groups, such as H, unsubstituted, or substituted with one or more R groups. c The following groups are substituted: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl.

[0014] Preferably, R8 is H; R9 is selected from H, unsubstituted, or substituted by one or more R... c The following groups are substituted: C 1-6 Alkyl, C 3-6 Cycloalkyl groups, preferably selected from H, unsubstituted, or substituted with one or more R groups. c Replacement C 1-6 Alkyl groups, such as H, unsubstituted, or substituted with one or more R groups. c The following groups are substituted: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl.

[0015] Preferably, each R c They may be the same or different, and are independently selected from hydroxyl, halogen, cyano, and C groups. 1-6 Alkyl, C 1-6 Alkyloxy, C 3-6 Cycloalkyl.

[0016] According to an embodiment of the present invention, R 10 Selected from H or C 1-6Alkyl groups, such as H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl.

[0017] According to an embodiment of the present invention, the compound of formula I has the structure shown in formula I-1;

[0018] According to an embodiment of the present invention, the compound of formula I is ozerfloxacin.

[0019] According to embodiments of the present invention, the pharmaceutically acceptable salt is selected from organic and inorganic salts of compounds of formula I (e.g., compounds of formula I-1). For example, the organic salt is selected from one or more of sulfonates, carboxylates, amino acid salts, and fatty acid salts of compounds of formula I (e.g., compounds of formula I-1), and the inorganic salt is selected from one or more of hydrochlorides, bromates, iodates, sulfates, hydrogen sulfates, phosphates, hydrogen phosphates, dihydrogen phosphates, and nitrates of compounds of formula I (e.g., compounds of formula I-1).

[0020] According to an embodiment of the present invention, the medicament is a pharmaceutical composition comprising a compound of formula I (e.g., a compound of formula I-1) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0021] According to embodiments of the present invention, the medicament or pharmaceutical composition contains a therapeutically effective amount of a compound of formula I (e.g., a compound of formula I-1) or a pharmaceutically acceptable salt thereof.

[0022] According to embodiments of the present invention, the pharmaceutically acceptable excipients include, but are not limited to, pharmaceutically acceptable carriers or excipients. For example, the pharmaceutically acceptable excipients are selected from at least one of the following: fillers, disintegrants, binders, lubricants, surfactants, flavoring agents, humectants, pH adjusters, solubilizers or cosolvents, osmotic pressure regulators, etc.

[0023] According to embodiments of the present invention, the pharmaceutical composition may further comprise at least one other antituberculosis drug. For example, the other antituberculosis drug is selected from one or more of isoniazid, rifampin, pyrazinamide, ethambutol, para-aminosalicylic acid, prothionamide, ethionamide, cycloserine, amikacin, capreomycin, rifapentine, rifabutin, fluoroquinolones (e.g., levofloxacin, moxifloxacin), kanamycin, streptomycin, bedaquiline, delamani, pretomani, etc., preferably selected from one or more of isoniazid, rifampin, ethambutol, and streptomycin.

[0024] According to embodiments of the present invention, the routes of administration of the drug include, but are not limited to, intravenous infusion, intravenous drip, subcutaneous administration, intradermal administration, intramuscular injection, oral spray, oral administration, and in situ administration to the tumor. According to embodiments of the present invention, the drug is preferably a pharmaceutical preparation, such as tablets, capsules, pills, granules, solutions, suspensions, syrups, injections (including injection solutions, sterile powders for injection, or concentrated solutions for injection), suppositories, inhalers, or sprays.

[0025] According to embodiments of the present invention, the drug is a single-dose formulation or a multi-dose formulation. The multi-dose formulation comprises more than one unit package, for example, consisting of 2 to 10 unit packages, such as 2, 3, 4, 5, 6, 7, 8, 9, or 10 unit packages. Each unit package contains the same amount of the compound of formula I and its pharmaceutically acceptable salts, or at least two unit packages contain different amounts of the compound of formula I and its pharmaceutically acceptable salts.

[0026] The present invention also provides a method for preventing and / or treating Mycobacterium tuberculosis infection, comprising administering to a patient a therapeutically effective amount of the compound of formula I (e.g., compound of formula I-1) or a pharmaceutically acceptable salt thereof.

[0027] The present invention also provides a method for preventing and / or treating Mycobacterium tuberculosis infection, comprising administering the treatment drug or drug composition to a patient.

[0028] According to an embodiment of the present invention, the Mycobacterium tuberculosis infection is selected from non-drug-resistant Mycobacterium tuberculosis infection and / or drug-resistant Mycobacterium tuberculosis infection, such as non-drug-resistant tuberculosis and / or drug-resistant tuberculosis.

[0029] According to an embodiment of the present invention, the non-drug-resistant Mycobacterium tuberculosis is the H37Rv standard strain.

[0030] According to an embodiment of the present invention, the drug-resistant Mycobacterium tuberculosis is a strain resistant to at least one known tuberculosis drug. As an example, the known tuberculosis drugs include, but are not limited to, one, two, three, or four of rifampin, isoniazid, moxifloxacin, and levofloxacin.

[0031] Preferably, the drug-resistant Mycobacterium tuberculosis is a Mycobacterium tuberculosis that is resistant to at least rifampin and isoniazid; for example, the drug-resistant Mycobacterium tuberculosis is a strain that is resistant to rifampin and isoniazid, and optionally resistant to at least one other known tuberculosis drug (e.g., fluoroquinolone drugs, including but not limited to moxifloxacin and levofloxacin).

[0032] According to an embodiment of the present invention, the drug-resistant tuberculosis is selected from at least one of the following: single-drug resistant tuberculosis (MR-TB), multidrug resistant tuberculosis (PR-TB), rifampicin resistant tuberculosis (RR-TB), isoniazid resistant tuberculosis (Hr-TB), multidrug resistant tuberculosis (MDR-TB), pre-extensively drug-resistant tuberculosis (Pre-XDR-TB), and extensively drug-resistant tuberculosis (XDR-TB).

[0033] According to an embodiment of the present invention, the tuberculosis is tuberculosis caused by infection with H37Rv standard strain and / or multidrug-resistant Mycobacterium tuberculosis strain.

[0034] Beneficial effects The present invention unexpectedly discovered that the quinolone compounds or their salts represented by Formula I have significant anti-tuberculosis activity, exhibiting excellent antibacterial activity against the H37Rv standard strain and drug-resistant Mycobacterium tuberculosis, with activity superior to the first-line drug isoniazid; secondly, it exhibits excellent antibacterial activity against Mycobacterium tuberculosis strains resistant to all fluoroquinolone drugs, indicating that its mechanism of action differs from that of fluoroquinolone drugs; furthermore, it has additive antibacterial activity with anti-tuberculosis drugs such as rifampin, isoniazid, ethambutol, and streptomycin, without antagonistic effects, and is expected to become an effective therapeutic drug for the prevention and treatment of Mycobacterium tuberculosis infectious diseases.

[0035] Terminology Definitions and Explanations Unless otherwise stated, the definitions of terms recorded in this application specification and claims, including definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, and definitions of specific compounds in the examples, can be arbitrarily combined and combined with each other. Such combinations and combinations shall fall within the scope of this application specification.

[0036] Unless otherwise stated, the numerical ranges described in this specification and claims are equivalent to describing at least each specific integer value therein. For example, the numerical range "1-12" is equivalent to describing each integer value in the numerical range "1-12", namely 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.

[0037] It should be understood that when describing one or more, or one or more types of items in this specification and claims, "a plurality of" or "multiple" means a quantity greater than or equal to 2, such as 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0038] Term "C" 1-12 "alkyl" should be understood to refer to straight-chain and branched alkyl groups having 1 to 12 carbon atoms, "C 1-6"Alkyl" means a straight-chain or branched alkyl group having 1, 2, 3, 4, 5, or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, or 1,2-dimethylbutyl, or their isomers.

[0039] Term "C" 3-12 "Cycloalkyl" should be understood to refer to saturated monovalent monocyclic, bicyclic (such as fused ring, bridged ring, spiro ring) hydrocarbon rings or tricyclic alkanes, having 3 to 12 carbon atoms, preferably "C". 3-8 cycloalkyl. The term "C" 3-12 "Cycloalkyl" should be understood to refer to a saturated monovalent monocyclic, bicyclic (e.g., bridged, spirocyclic) hydrocarbon ring or tricyclic alkane having 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbon atoms. The C... 3-12 Cycloalkyl groups can be monocyclic hydrocarbon groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or cyclodecyl; or bicyclic hydrocarbon groups, such as borneolyl, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonyl, 2,6-diazaspiro[3,4]octyl; or tricyclic hydrocarbon groups, such as adamantyl.

[0040] The term "alkoxy" refers to -O- (alkyl), where alkyl is defined as described above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, and butoxy.

[0041] The term "halogen" includes F, Cl, Br, and I.

[0042] The term "pharmaceuticalally acceptable excipient" refers to an excipient that does not cause significant irritation to the organism and does not impair the biological activity and properties of the active compound.

[0043] Unless otherwise stated, the Formula I compounds and / or their pharmaceutically acceptable salts described in this specification may include their tautomers, amorphous forms, polymorphs, solvates, isotope-labeled forms (such as deuterated derivatives), etc.

[0044] The term "tautomer" refers to a functional group isomer resulting from the rapid movement of an atom between two positions within a molecule. The compounds disclosed herein can exhibit tautomerism. Tautomers can exist in two or more interconvertible forms. Proton-transfer tautomers arise from the migration of covalently bonded hydrogen atoms between two atoms. Tautomers generally exist in equilibrium form, and attempts to isolate a single tautomer typically yield a mixture whose physicochemical properties are consistent with those of the mixture of compounds. The equilibrium position depends on the intramolecular chemical characteristics. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form is dominant; while in phenols, the enol form is dominant. This disclosure encompasses all tautomeric forms of the compounds.

[0045] In this application, the term "solvent" refers to a compound of the present disclosure or a salt thereof comprising a stoichiometric or non-stoichiometric solvent bound by intermolecular noncovalent forces, and a hydrate when the solvent is water.

[0046] "Isotope" refers to all isotopes of atoms appearing in the compounds of this invention. Isotopes include those atoms having the same atomic number but different mass numbers. Examples of isotopes suitable for inclusion in the compounds of this invention are hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, respectively, for example, but not limited to, [examples of isotopes]. 2 H, 3 H, 13 C 14 C 15 N、 18 O、 31 P, 32 P, 35 S, 18 F and 36 C1. The isotope-labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art or by methods similar to those described in the appended examples, using suitable isotope-labeled reagents instead of non-isotope-labeled preparations. Such compounds have a variety of potential uses, for example, as standards and reagents in the determination of biological activity. In the case of stable isotopes, such compounds have the potential to advantageously alter biological, pharmacological, or pharmacokinetic properties.

[0047] The term "H37Rv standard strain" refers to the most commonly used standard viral strain of Mycobacterium tuberculosis in current research on pulmonary tuberculosis.

[0048] The term "drug-resistant tuberculosis (MR-TB)" refers to resistance to a single first-line anti-tuberculosis drug. The term "multidrug-resistant tuberculosis (PR-TB)" refers to resistance to more than one first-line anti-tuberculosis drug, but does not include simultaneous resistance to isoniazid and rifampin. The term "rifampicin-resistant tuberculosis (RR-TB)" refers to resistance to rifampicin, regardless of whether it is resistant to other anti-tuberculosis drugs. The term "isoniazid-resistant tuberculosis (Hr-TB)" refers to tuberculosis that is resistant to isoniazid but sensitive to rifampin. The term "multidrug-resistant tuberculosis (MDR-TB)" refers to resistance to at least two first-line anti-tuberculosis drugs, including simultaneous resistance to isoniazid and rifampin. The term "pre-extensively drug-resistant tuberculosis (Pre-XDR-TB)" is defined as MDR-TB / RR-TB, which is resistant to any one of the fluoroquinolone drugs. The term “extensively drug-resistant tuberculosis (XDR-TB)” is defined as MDR-TB / RR-TB, which is resistant to any one fluoroquinolone drug and at least one other Group A drug (bedaquiline or linezolid).

[0049] The term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, with humans being the most preferred.

[0050] The term “therapeutic effective dose” refers to the amount of an active compound or drug that researchers, veterinarians, physicians, or other clinicians are searching for in a tissue, system, animal, individual, or human to elicit a biological or medical response. It includes one or more of the following: (1) prevention of disease: e.g., prevention of disease, disorder, or condition in an individual who is susceptible to disease, disorder, or symptom but has not yet experienced or developed the pathology or symptoms of the disease; (2) inhibition of disease: e.g., inhibition of disease, disorder, or symptom in an individual experiencing or developing the pathology or symptoms of the disease (i.e., prevention of further development of the pathology and / or symptoms); (3) relief of disease: e.g., relief of disease, disorder, or symptom in an individual experiencing or developing the pathology or symptoms of the disease (i.e., reversal of the pathology and / or symptom). The specific dosage will vary depending on factors such as the specific compound chosen, the dosing regimen, whether it is administered in combination with other compounds, the timing of administration, the tissue to which the drug is administered, and the physical delivery system used. Attached Figure Description

[0051] Figure 1 Compound I-1 1 H NMR spectrum; Figure 2 Liquid chromatogram of compound I-1; Figure 3 Time-kill curves of bactericidal activity of different concentrations of ozenocin against H37Rv and MDR strains. Detailed Implementation

[0052] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0053] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0054] In the following embodiments: The compound to be tested, I-1, is named ozelaxacin. It is sourced from TargetMol Chemicals Inc., manufacturer number T7902, and has a purity of 99.33%. 1 H NMR spectrum as follows Figure 1 The liquid chromatogram is as follows Figure 2 .

[0055] Strains were sourced from the following strains: H37Rv standard strain: provided by Guangdong Provincial Tuberculosis Control Center; multidrug-resistant Mycobacterium tuberculosis strains (resistant to isoniazid, rifampin, moxifloxacin, and levofloxacin): clinical isolates from the Third People's Hospital of Zhuhai.

[0056] Example 1: Evaluation of the anti-tuberculosis effect of ozenafloxacin Strains: H37Rv standard strain, multidrug-resistant Mycobacterium tuberculosis strain (resistant to isoniazid, rifampin, moxifloxacin and levofloxacin).

[0057] Experimental Materials and Methods: The microplate alamar blue assay (MABA) was used to evaluate the in vitro antibacterial activity of compounds against H37Rv standard strain and multidrug-resistant Mycobacterium tuberculosis strain. The MABA experiment was performed as follows: (1) Culture medium preparation: Middlebrook 7H9 culture medium was prepared and 0.05% Tween-80, 0.2% glycerol and 10% albumin-glucose-catalase supplement (BBL Middlebrook ADC Enrichment, BectonDickinson) were added to promote the growth of the strain.

[0058] (2) Inoculation of strains: The H37Rv standard strain and the multidrug-resistant tuberculosis mycobacterium strain cultured for 2 to 3 weeks were inoculated into the above culture medium and adjusted to the McFarland 1 standard concentration (approximately 2 × 10⁻⁶). 8 (CFU / mL), to be used as a bacterial suspension for experiments.

[0059] (3) Culture medium preparation: Prepare 7H9 culture medium without Tween-80 and add it to a sterile 96-well microtiter plate, 100 µL per well.

[0060] (4) Compound dissolution and dilution: The test compounds were first dissolved in DMSO and diluted with pure water to 128 μg / mL. Then, these compounds were serially diluted twofold using 7H9 medium without Tween-80, and 100 µL of each solution was added to a sterile 96-well plate. The final drug concentration range was 0.25–32 μg / mL. Isoniazid was used as a positive control, and the final concentration of DMSO in all wells was ensured to be ≤ 0.1% to minimize interference with the experimental results.

[0061] (5) Inoculation and incubation: Dilute the McFarland 1 standard concentration bacterial suspension at a ratio of 1:20. Add 100 µL of the diluted bacterial suspension to each well, with a final volume of 200 µL per well and a final bacterial suspension concentration of 5 × 10⁻⁶. 6 CFU / mL. Place sterile 96-well plates in an incubator and incubate at 37°C for 8 days.

[0062] (6) Alamar Blue detection: After the incubation period, add 20 µL of 10× Alamar Blue and 50 µL of 10% Tween-80 to each well, place them in a constant temperature CO2 incubator, and incubate at 37°C for 24 hours in a 5% CO2 environment.

[0063] (7) Fluorescence detection: After 24 hours of incubation, fluorescence microplate readers were used to detect the fluorescence at an excitation wavelength of 560 nm and an emission wavelength of 590 nm. The corresponding fluorescence intensity (Fluorescence units, FU) was recorded to evaluate the inhibitory effect of the compound.

[0064] (8) Determination of minimum inhibitory concentration (MIC): MIC is defined as the lowest concentration of compound that produces at least 90% inhibition. A positive control group containing the H37Rv standard strain without antibiotics was set up to monitor bacterial growth; a negative control group was set up to 7H9 medium without Tween-80 to ensure sterility of the experiment.

[0065] The experimental results are shown in Table 1.

[0066] Table 1. Antibacterial activity of ozenafloxacin against H37Rv standard strain and multidrug-resistant Mycobacterium tuberculosis strain in vitro.

[0067] aMultidrug-resistant Mycobacterium tuberculosis: resistant to isoniazid, rifampin, moxifloxacin and levofloxacin.

[0068] The results showed that ozenafloxacin has a significant antibacterial effect against Mycobacterium tuberculosis. The MIC for both the H37Rv standard strain and multidrug-resistant Mycobacterium tuberculosis was 0.125 μg / mL, exceeding that of the positive control drug isoniazid, providing a new and feasible treatment option for tuberculosis. Ozefloxacin is a quinolone drug, and its mechanism of action differs from that of the fluoroquinolones moxifloxacin and levofloxacin.

[0069] Example 2: Evaluation of the synergistic antibacterial effect of ozenafloxacin with existing anti-tuberculosis drugs in vitro Strains: H37Rv standard strain.

[0070] Experimental Materials and Methods: Drug interactions were assessed using a checkerboard titration method to compare the candidate drug ozenafil with existing anti-tuberculosis drugs (rifampin, isoniazid, ethambutol, and streptomycin) via microdilution. The specific steps are as follows: (1) In a 96-well microtiter plate, the drugs were diluted in 100 µL volumes. Compound I-1, ozenafil, was serially diluted vertically in two-fold increments (rows A to G), and rifampin, isoniazid, ethambutol, or streptomycin was serially diluted horizontally in two-fold increments (columns 1 to 10) to obtain various combinations of the two drugs. The bottom row (row H) and column 11 contained only a single drug, and column 12 was a drug-free control.

[0071] (2) Add 100 µL of H37Rv standard strain culture medium in logarithmic growth phase (dilution of McFarland 1 standard concentration bacterial suspension at a ratio of 1:20) to each well. Place the plate in a sealed bag and incubate at 37°C for 2 weeks, then read the results. The lowest drug concentration corresponding to the well that produces at least 90% inhibition is considered the minimum inhibitory concentration (MIC).

[0072] (3) Synergy is determined by calculating the fractional inhibitory concentration (FIC) of ozenacin, which is the sum of the fractional inhibitory concentrations of rifampin, isoniazid, ethambutol, or streptomycin. The formula for calculating the fractional inhibitory concentration (FIC) is as follows:

[0073] ΣFIC ≤ 0.5 indicates a synergistic effect, ΣFIC ≥ 4.0 indicates an antagonistic effect, and values ​​in between indicate an additive effect.

[0074] The experimental results are shown in Table 2.

[0075] Table 2. Ozefloxacin and its comparison with rifampin, isoniazid, ethambutol, and streptomycin as anti-tuberculosis drugs. Synergistic antibacterial activity against H37Rv standard strain in vitro

[0076] The results showed that ozenafloxacin had an additive antibacterial effect in vitro with the anti-tuberculosis drugs rifampin, isoniazid, ethambutol, and streptomycin.

[0077] Example 3: Evaluation of the time-bactericidal activity of ozenafloxacin against H37Rv and MDR strains using time-bactericidal curves Strains: H37Rv standard strain, multidrug-resistant Mycobacterium tuberculosis strain (resistant to isoniazid, rifampin, moxifloxacin and levofloxacin).

[0078] Experimental Materials and Methods: This study used the YK-320 system and its matching mycobacterial liquid culture tubes. The culture tubes mainly contained 7H9 medium and ADC additives, and detection was based on fluorescence. The experiment was conducted at a constant temperature of 37 ℃, with a culture and analysis cycle set at 360 hours. The system excited fluorescence signals at a wavelength of 560 nm per hour and automatically acquired data. Both the experimental and control groups had culture tubes prepared with a final concentration of 10... 5 CFU / mL bacterial suspension. Different concentrations of ozenacin (OZN) were added to the experimental groups, with final concentrations of 0.5 µg / mL, 0.25 µg / mL, and 0.125 µg / mL, respectively; a control group without the drug was also set up as a control.

[0079] The results showed that ozetin exhibited strong, concentration-dependent inhibitory activity against both Mycobacterium tuberculosis H37Rv and MDR strains. For H37Rv strain: 0.25 µg / mL and 0.5 µg / mL ozetin completely inhibited bacterial growth over a 360-hour detection period; 0.125 µg / mL delayed its logarithmic growth phase by approximately 129 hours. Figure 3 (A). The inhibitory effect of ozenafloxacin on MDR strains was also significant, with concentrations of 0.125 µg / mL, 0.25 µg / mL, and 0.5 µg / mL delaying its logarithmic growth phase by approximately 34 hours, 94 hours, and 186 hours, respectively. Figure 3 (Middle B). The 360-hour long-term culture used in this study, compared with the conventional 168-hour culture, more fully revealed the sustained inhibitory effect of ozenafloxacin on slow-growing Mycobacterium tuberculosis.

[0080] In summary, ozenafloxacin exhibits significant anti-tuberculosis activity in vitro, demonstrating excellent antibacterial activity against both the H37Rv standard strain and multidrug-resistant Mycobacterium tuberculosis. Furthermore, it shows additive antibacterial activity in vitro with first-line anti-tuberculosis drugs such as rifampin, isoniazid, and ethambutol, without antagonistic effects, making it a promising treatment for Mycobacterium tuberculosis infections. This invention lays a scientific experimental foundation for expanding the clinical indications of ozenafloxacin.

[0081] The above description provides an exemplary account of the implementation methods of the technical solution disclosed herein. It should be understood that the scope of protection of this disclosure is not limited to the above-described embodiments. Any modifications, equivalent substitutions, or improvements made by those skilled in the art within the spirit and principles of this disclosure should be included within the scope of protection of the claims of this application.

Claims

1. Use of a compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of a medicament, wherein the medicament is used for the prevention and / or treatment of Mycobacterium tuberculosis infection: in, R1 is selected from those without substitution or by one or more R1s. a Replacement C 3-12 cycloalkyl, wherein each R a They may be the same or different, and are independently selected from hydroxyl, halogen, cyano, and C groups. 1-12 Alkyl, C 1-12 Alkyloxy; R2 and R7 may be the same or different, and are independently selected from those without substitution or by one or more R... b Replacement C 1-12 Alkyl groups, wherein each R b They may be the same or different, and are independently selected from hydroxyl, halogen, cyano, and C groups. 1-12 Alkyl, C 1-12 Alkyloxy, C 3-12 cycloalkyl; R3, R4, R5, and R6 may be the same or different, and are independently selected from H, hydroxyl, halogen, cyano, and C. 1-12 Alkyl, C 1-12 Alkyloxy, C 3-12 cycloalkyl; R8 and R9 may be the same or different, and are independently selected from H, unsubstituted, or by one or more R... c The following groups are substituted: C 1-12 Alkyl, C 3-12 cycloalkyl, wherein each R c They may be the same or different, and are independently selected from hydroxyl, halogen, cyano, and C groups. 1-12 Alkyl, C 1-12 Alkyloxy, C 3-8 cycloalkyl; R 10 Selected from H or C 1-12 alkyl; Preferably, the Mycobacterium tuberculosis infection is selected from non-drug-resistant Mycobacterium tuberculosis infection and / or drug-resistant Mycobacterium tuberculosis infection, such as non-drug-resistant tuberculosis and / or drug-resistant tuberculosis.

2. The use according to claim 1, wherein: R1 is selected from those without substitution or by one or more R1s. a Replacement C 3-6 Cycloalkyl, each R a They may be the same or different, and are independently selected from hydroxyl, halogen, cyano, and C groups. 1-6 Alkyl, C 1-6 Alkyloxy; R2 and R7 may be the same or different, and are independently selected from those without substitution or by one or more R... b Replacement C 1-6 Alkyl, each R b They may be the same or different, and are independently selected from hydroxyl, halogen, cyano, and C groups. 1-6 Alkyl, C 1-6 Alkyloxy, C 3-6 cycloalkyl; R3, R4, R5, and R6 may be the same or different, and are independently selected from H, hydroxyl, halogen, cyano, and C. 1-6 Alkyl, C 1-6 Alkyloxy, C 3-6 cycloalkyl; R8 and R9 may be the same or different, and are independently selected from H, unsubstituted, or by one or more R... c The following groups are substituted: C 1-6 Alkyl, C 3-6 Cycloalkyl, each R c They may be the same or different, and are independently selected from hydroxyl, halogen, cyano, and C groups. 1-6 Alkyl, C 1-6 Alkyloxy, C 3-6 cycloalkyl; Preferably, R8 is H; R9 is selected from H, unsubstituted, or substituted by one or more R... c The following groups are substituted: C 1-6 Alkyl, C 3-6 cycloalkyl; R 10 Selected from H or C 1-6 Alkyl groups, such as H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl.

3. The use according to claim 1, wherein the compound of formula I has the structure shown in formula I-1; 。 4. The use according to any one of claims 1-3, wherein the drug is a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable excipient present or absent.

5. The use according to claim 4, wherein the pharmaceutical composition may further comprise at least one other anti-tuberculosis drug; For example, the other anti-tuberculosis drugs are selected from one or more of isoniazid, rifampin, pyrazinamide, ethambutol, para-aminosalicylic acid, prothionamide, ethionamide, cycloserine, amikacin, capreomycin, rifapentine, rifabutin, fluoroquinolones (e.g., levofloxacin, moxifloxacin), kanamycin, streptomycin, bedaquiline, delamani, pretomani, etc., preferably selected from one or more of isoniazid, rifampin, ethambutol, and streptomycin.

6. The use according to any one of claims 1-5, wherein the Mycobacterium tuberculosis infection is selected from non-drug-resistant Mycobacterium tuberculosis infection and / or drug-resistant Mycobacterium tuberculosis infection.

7. The use according to claim 6, wherein: The non-drug-resistant Mycobacterium tuberculosis strain is the H37Rv standard strain; The drug-resistant Mycobacterium tuberculosis is a strain resistant to at least one known tuberculosis drug; for example, the known tuberculosis drug includes, but is not selected from, 1, 2, 3 or 4 of rifampin, isoniazid, moxifloxacin and levofloxacin.

8. The use according to claim 7, wherein: The drug-resistant Mycobacterium tuberculosis is Mycobacterium tuberculosis that is resistant to at least rifampin and isoniazid; For example, the drug-resistant Mycobacterium tuberculosis is a strain that is resistant to rifampin and isoniazid, and optionally resistant to at least one other known tuberculosis drug (e.g., fluoroquinolones, including but not limited to moxifloxacin and levofloxacin).

9. The use according to claim 6, wherein the Mycobacterium tuberculosis infection is non-drug-resistant tuberculosis and / or drug-resistant tuberculosis.

10. The use according to claim 9, wherein the drug-resistant tuberculosis is selected from at least one of the following: single-drug resistant tuberculosis (MR-TB), multidrug resistant tuberculosis (PR-TB), rifampicin resistant tuberculosis (RR-TB), isoniazid resistant tuberculosis (Hr-TB), multidrug resistant tuberculosis (MDR-TB), pre-extensively drug-resistant tuberculosis (Pre-XDR-TB), and extensively drug-resistant tuberculosis (XDR-TB).

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