Use of a compound in the preparation of mycobacterial inhibitors

Abstract

This invention provides a compound of formula (I), its stereoisomers, its optical isomers, its pharmaceutically acceptable salts, its crystal form, its isotopic derivatives, its prodrug, its metabolites, its solvates, or its hydrates for the preparation of pharmaceutical compositions for the treatment and / or prevention of diseases associated with Mycobacterium tuberculosis and / or non-tuberculous mycobacteria. The compounds of this invention exhibit excellent bactericidal effects and high specificity, showing no cross-resistance with existing anti-tuberculosis drugs, thus providing a breakthrough direction for the treatment of drug-resistant tuberculosis.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to the use of a compound in the preparation of mycobacterial inhibitors. Background Technology

[0002] Tuberculosis (TB) is a chronic infectious disease caused by Mycobacterium tuberculosis (Mtb), and is one of the deadliest diseases in the world (Med Res Rev, 2020, 40(1):263-292). The four first-line anti-tuberculosis drugs (rifampin, isoniazid, pyrazinamide, and ethambutol) and eight second-line anti-tuberculosis drugs (streptomycin, ethionamide, para-aminosalicylic acid, capreomycin, kanamycin, ofloxacin, amikacin, and cycloserine) commonly used in clinical practice were all discovered between the 1940s and 1970s.

[0003] In order to shorten the treatment time, improve the efficacy, and reduce the chance of drug-resistant tuberculosis, the WHO recommends a treatment regimen that combines multiple drugs. However, this still cannot reduce the occurrence of drug resistance. Therefore, new anti-tuberculosis drugs are urgently needed.

[0004] Mycolic acid is a major component of the outermost layer of the mycobacterial cell wall, accounting for 60% of the cell's dry weight. Mycolic acid is not only an important pathogenic substance in mycobacteria, but its metabolic pathway is also a specific target of anti-tuberculosis drugs. For example, anti-tuberculosis drugs such as isoniazid (INH) target the mycolic acid synthesis pathway (Journal of Bacteriology, 2000, 182(14):4059; Microbiology, 2000, 146(2):289-296.). InhA is a key enzyme in the mycolic acid synthesis pathway of mycobacteria, and mycolic acid is an important component of the Mycobacterium tuberculosis cell wall, directly affecting the bacteria's survival ability and drug resistance.

[0005] INH is the most well-known first-line treatment inhibitor of InhA. However, since INH is a prodrug, mutations in both the InhA and KatG genes can lead to drug resistance. Therefore, researchers are currently working to discover novel potential inhibitors of Mycobacterium tuberculosis that can directly inhibit the catalytic activity of the InhA enzyme without requiring an activation phase. Summary of the Invention

[0006] To address the aforementioned problems, this invention provides a novel mycobacterial inhibitor targeting InhA, a key enzyme in the mycolic acid synthesis pathway of mycobacteria.

[0007] In a first aspect of the invention, there is provided the use of a compound of formula (I), its stereoisomers, its optical isomers, its pharmaceutically acceptable salts, its crystal forms, its isotopic derivatives, its prodrugs, its metabolites, its solvates, or its hydrates thereof, for the preparation of pharmaceutical compositions for treating and / or preventing diseases associated with Mycobacterium tuberculosis and / or non-tuberculous mycobacteria.

[0008]

[0009] Among them, R1, R2, and R3 are each independently selected from the following groups: H, D, C 1-4 Alkyl, C 1-4 Deuterated alkyl groups.

[0010] In another preferred embodiment, the nontuberculous mycobacteria include Mycobacterium abscessus.

[0011] In a preferred embodiment, the compound is selected from the group consisting of:

[0012]

[0013] In a preferred embodiment, the disease associated with Mycobacterium tuberculosis is tuberculosis; the disease associated with nontuberculous mycobacteria is nontuberculous mycobacterial disease.

[0014] Preferably, the tuberculosis is selected from the group consisting of: pulmonary tuberculosis, renal tuberculosis, bladder tuberculosis, reproductive system tuberculosis, intestinal tuberculosis, tuberculous peritonitis, bone and joint tuberculosis, tuberculous meningitis, lymph node tuberculosis, cutaneous tuberculosis, and hematogenous disseminated tuberculosis;

[0015] The nontuberculous mycobacterial diseases mentioned are selected from the following group: lung infections, skin and soft tissue infections, and lymphadenitis.

[0016] In a preferred embodiment, the compound reduces the viable count of standard strains of Mycobacterium tuberculosis and / or non-tuberculous mycobacteria by ≥1.5 log₂ / ₅ at a concentration of 2 × MIC within 9 days (e.g., 7, 6, 5, 4, 3, or 2 days). 10 Preferably ≥2log 10 More preferably ≥2.5log 10 .

[0017] In another preferred embodiment, the compound reduces the viable count of a standard strain of Mycobacterium tuberculosis by ≥3 log₂ within 9 days (e.g., 7, 6, 5, 4, 3, or 2 days) at a concentration of 2 × MIC. 10 Preferably ≥3.5log 10 More preferably ≥4log 10 .

[0018] In another preferred embodiment, the compound reduces the viable count of a standard strain of Mycobacterium tuberculosis by ≥3 log within 9 days (e.g., 7, 6, 5, 4, 3, or 2 days) at a concentration of 10 × MIC. 10 Preferably ≥3.5log 10 More preferably ≥4log 10 The optimal value is ≥5log 10 The optimal value is ≥5.5log. 10 .

[0019] In another preferred embodiment, the standard strain of Mycobacterium tuberculosis is H37Rv.

[0020] In another preferred embodiment, the compound reduces the viable count of a standard strain of nontuberculous mycobacteria by ≥3 log₂ within 9 days (e.g., 7, 6, 5, 4, 3, or 2 days) at a concentration of 2 × MIC. 10 In another preferred embodiment, the compound reduces the viable count of a standard strain of nontuberculous mycobacteria by ≥3 log at a concentration of 10 × MIC for 9 days (e.g., 7, 6, 5, 4, 3, or 2 days). 10 Preferably ≥3.5log 10 More preferably ≥4log 10 The optimal value is ≥5log 10 The optimal value is ≥5.5log. 10 .

[0021] In another preferred embodiment, the standard strain of the nontuberculous mycobacterium is ATCC 19977.

[0022] In a preferred embodiment, the compound has no antibacterial activity (MIC) against Salmonella, Enterobacter cloacae, Klebsiella pneumoniae, and / or Staphylococcus aureus. 90 >1000μm).

[0023] In another preferred embodiment, the compound exhibits an antimicrobial MIC against standard strains of Mycobacterium tuberculosis and / or Mycobacterium abscessis. 90 The range is 30–260 μm, for example, 31.25, 62.5, 125 or 250 μm.

[0024] In another preferred embodiment, the compound has an antimicrobial MIC against widely resistant strains of Mycobacterium tuberculosis. 90 The range is 30–260 μm, for example, 31.25, 62.5, 125 or 250 μm.

[0025] In another preferred embodiment, the broadly drug-resistant strains of Mycobacterium tuberculosis are selected from the group consisting of:

[0026] XDR-1 is resistant to isoniazid, rifampin, streptomycin, ethambutol, ofloxacin, levofloxacin, rifabutin, pyrazinamide, and para-aminosalicylic acid.

[0027] XDR-2 is resistant to isoniazid, rifampin, streptomycin, ethambutol, ofloxacin, levofloxacin, rifabutin, pyrazinamide, kanamycin, capreomycin, and amikacin.

[0028] XDR-3 is resistant to isoniazid, rifampin, streptomycin, ethambutol, ofloxacin, levofloxacin, rifabutin, pyrazinamide, kanamycin, capreomycin, amikacin, and para-aminosalicylic acid.

[0029] In another preferred embodiment, the compound has an antimicrobial MIC against clinically susceptible strains of Mycobacterium tuberculosis. 90 All range from 30 to 260 μm, for example, 31.25, 62.5, 125 or 250 μm.

[0030] In another preferred embodiment, the compound reduces the viable number of dormant Mycobacterium tuberculosis bacteria at a concentration of 2×MIC by ≥2log over 24 days (e.g., 22, 20, 17, 14, 12, 10, 8, 6, 4, 3, or 2 days). 10 3log is preferred. 10 More preferably ≥4log 10 The optimal value is ≥5log 10 The optimal value is ≥5.5log. 10 .

[0031] In another preferred embodiment, the compound reduces the viable number of dormant Mycobacterium tuberculosis bacteria at a concentration of 10×MIC by ≥2log over 24 days (e.g., 22, 20, 17, 14, 12, 10, 8, 6, 4, 3, or 2 days). 10 3log is preferred. 10 More preferably ≥4log 10 The optimal value is ≥5log 10 The optimal value is ≥5.5log. 10 .

[0032] In another preferred embodiment, the compound reduces the viable count of dormant nontuberculous mycobacteria by ≥3 log within 12 days (e.g., 10, 9, 8, 6, 4, 3, or 2 days) at a concentration of 2 × MIC. 10 Preferably ≥4log 10 More preferably ≥5log 10 The optimal value is ≥5.5log. 10 The optimal choice is ≥6log 10 .

[0033] In another preferred embodiment, the compound reduces the viable count of dormant nontuberculous mycobacteria by ≥3 log₂ within 12 days (e.g., 10, 9, 8, 6, 4, 3, or 2 days) at a concentration of 10 × MIC. 10 Preferably ≥4log 10 More preferably ≥5log 10 The optimal value is ≥5.5log. 10 The optimal choice is ≥6log 10 .

[0034] In a preferred embodiment, the compound inhibits the activity of InhA.

[0035] In a preferred embodiment, the pharmaceutical composition further includes other anti-tuberculosis mycobacterial drugs and / or other anti-nontuberculosis mycobacterial drugs.

[0036] In a preferred embodiment, the other anti-tuberculosis mycobacterial drugs are selected from the group consisting of: rifampin, isoniazid, pyrazinamide, ethambutol, streptomycin, ethionamide, para-aminosalicylic acid, capreomycin, kanamycin, ofloxacin, amikacin, cycloserine, bedaquiline, diramani, or combinations thereof;

[0037] The other anti-nontuberculous mycobacterial drugs mentioned are selected from the group consisting of: amikacin, clofazimine, azithromycin, clarithromycin, cefoxitin, moxifloxacin, ciprofloxacin, imipenem, meropenem, or combinations thereof.

[0038] In a preferred embodiment, the FICI of the compound of formula (I) with other anti-tuberculous mycobacterial drugs and / or anti-nontuberculous mycobacterial drugs is ≤1 and >0.5, preferably FICI ≤0.5.

[0039] In a preferred embodiment, the pharmaceutical composition further includes a pharmaceutically acceptable carrier, excipient, or excipient.

[0040] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0041] Figure 1 The in vitro bactericidal activity of compounds X-376 and X-396 in Example 4 is shown, where CFU represents the number of viable bacteria, and the test strain is the standard strain H37Rv of Mycobacterium tuberculosis.

[0042] Figure 2The in vitro bactericidal activity of compounds X-376 and X-396 in Example 4 is shown, where CFU represents the number of viable bacteria, and the test strain is the standard strain ATCC 19977 of Mycobacterium abscessus.

[0043] Figure 3 The ability of compounds X-376 and X-396 in Example 5 to eliminate dormant bacteria in vitro is shown, where CFU represents the number of viable bacteria, and the test strain is the standard strain H37Rv of Mycobacterium tuberculosis.

[0044] Figure 4 The ability of compounds X-376 and X-396 in Example 5 to eliminate dormant bacteria in vitro is shown, where CFU represents the number of viable bacteria, and the test strain is the standard strain ATCC 19977 of Mycobacterium abscessus. Detailed Implementation

[0045] Through long-term and in-depth research and extensive screening, the inventors have developed a novel mycobacterial inhibitor targeting InhA, a key enzyme in the mycolic acid synthesis pathway. The inventors previously evaluated the antibacterial efficacy, cross-resistance, and combination therapy with first-line anti-tuberculosis drugs using standard and clinical strains. They also plan to evaluate the therapeutic efficacy of these compounds against Mycobacterium tuberculosis using mouse models. Furthermore, they will utilize multi-omics and high-throughput transposon library screening methods to reveal the antibacterial mechanisms and bacterial response characteristics of these compounds. Finally, they will optimize the structure of the compounds to develop more effective drug-grade compounds. Based on these findings, the inventors have completed this invention.

[0046] Active ingredients

[0047] As used herein, “compound of the present invention” means a compound of formula (I), and also includes its stereoisomers, its optical isomers, its pharmaceutically acceptable salts, its crystal forms, its isotopic derivatives, its prodrugs, its metabolites, its solvates or hydrates thereof.

[0048] Unless otherwise specified, the structural formulas described in this invention are intended to include all stereoisomers (such as cis-trans isomers, enantiomers, diastereomers, and conformational isomers): R and S configurations containing an asymmetric center, (Z) and (E) isomers of double bonds, cis-trans isomers of cycloalkanes, etc. Therefore, any single stereochemical isomer of the compounds of this invention, or a mixture of its enantiomers, diastereomers, or conformational isomers, is within the scope of this invention.

[0049] The compounds of this invention may contain cis-trans isomers, one or more chiral carbon atoms, and thus can produce cis-trans isomers, chiral isomers, enantiomers, diastereomers, and other combinations of stereoisomers. Cis-trans isomerism refers to the diastereomeric phenomenon in which different functional groups in a compound molecule are arranged differently in space due to a restrictive factor that limits free rotation. This restrictive factor is generally caused by non-rotating functional groups in the structure of organic compounds, such as C=C double bonds, C=N double bonds, C=S double bonds, N=N double bonds, heterocycles, or cycloalkanes. Organic molecules containing such isomers, such as alkenes, azo compounds, and cycloalkanes, are considered cis-trans isomers. Cis refers to the same ligands being in adjacent positions, generally denoted by "cis" or "cis-"; trans refers to the same ligands being in diagonal positions, generally denoted by "trans" or "trans-". Each chiral carbon atom can be defined as (R)- or (S)- based on stereochemistry. This invention aims to include all possible isomers, their racemic and optically pure forms. The compounds of this invention can be prepared using racemic, cis-trans, chiral, diastereomer, or enantiomers as starting materials or intermediates. Optically active isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as crystallization and chiral chromatography.

[0050] Conventional techniques for preparing / separating individual optical isomers (i.e., cis-trans isomers and chiral isomers) include chiral synthesis from suitable cis-trans precursors or optically pure precursors, or resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high-performance liquid chromatography.

[0051] To design the synthesis of a specific stereoisomer of the compound of this invention, it can be prepared asymmetrically or derivatized with a chiral auxiliary. The resulting stereo mixture is then separated, and the chiral auxiliary is removed to obtain pure cis-trans monomers, chiral monomers, or mixed stereoisomers. If the molecule contains a cis-trans isomer center, it can be purified by column chromatography (normal-phase silica gel column or reverse-phase high-performance liquid chromatography) to obtain pure cis or trans products. Alternatively, if the molecule contains a basic functional group, such as an amino acid, or an acidic functional group, such as a carboxyl group, it can be formed with a suitable optically active acid or base to form a diastereomeric salt, which is then separated by conventional methods such as separation crystallization or chromatography to obtain pure enantiomers.

[0052] This invention also includes isotopically labeled compounds (i.e., isotopic derivatives), equivalent to the original compounds disclosed herein. However, it is common practice for one or more atoms to be substituted with atoms of different atomic weights or mass numbers. Examples of isotopes in the isotopic derivatives of this invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine isotopes, such as 2H, 3H, 13C, 11C, 14C, 15N, 18O, 17O, 31P, 32P, 35S, 18F, and 36Cl, respectively. Isotopic derivatives of the compounds of this invention are all within the scope of protection of this invention. In this document, 3H-labeled and 14C-labeled compounds are useful in tissue distribution experiments of drugs and substrates. Tritium (i.e., 3H) and carbon-14 (i.e., 14C)-labeled compounds are relatively easy to prepare and detect, and are the preferred isotopes. Furthermore, heavier isotope substitutions, such as deuterium (2H), are preferred in certain cases due to their excellent metabolic stability, which offers advantages in some therapies, such as increasing half-life or reducing dosage in vivo. Isotope-labeled compounds can be prepared using general methods by replacing non-isotopic reagents with readily available isotope-labeling reagents, according to the schemes disclosed in the examples.

[0053] As used herein, the term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0054] "Pharmaceutically acceptable acid addition salts" refer to salts formed with inorganic or organic acids that retain the bioavailability of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochlorides, hydrobroms, sulfates, nitrates, and phosphates; organic acid salts include, but are not limited to, formates, acetates, 2,2-dichloroacetate, trifluoroacetate, propionates, hexanoates, octanoates, decanoates, undecenoates, glycolates, gluconates, lactates, sebates, adipates, glutarate, malonates, oxalates, maleates, succinates, fumarates, tartrates, citrates, palmitates, stearates, oleates, cinnamates, laurates, malates, glutamates, pyroglutamates, aspartate, benzoates, methanesulfonates, benzenesulfonates, p-toluenesulfonates, alginates, ascorbic acid salts, salicylates, 4-aminosalicylic acid salts, and naphthalene disulfonates. These salts can be prepared using methods known in this field.

[0055] "Pharmaceutically acceptable base addition salts" refer to salts formed with inorganic or organic bases that retain the bioavailability of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Preferred inorganic salts are ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, the following: primary amines, secondary amines, and tertiary amines; substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. These salts can be prepared by methods known in the art.

[0056] Metabolites of the compound represented by formula (I) and its pharmaceutically acceptable salts, as well as prodrugs that can be converted in vivo into the compound represented by formula (I) and its pharmaceutically acceptable salts, are also included within the scope of protection of this invention.

[0057] As used herein, the term "solvent" refers to a complex of a compound of formula (I) coordinated with a solvent molecule in a specific ratio.

[0058] As used herein, the term "hydrate" refers to a complex of the compound represented by formula (I) coordinated with water molecules in a specific ratio.

[0059] As described herein, the compounds of the present invention can be substituted with any number of substituents or functional groups to broaden their scope. Generally, the term "substitution" refers to replacing a hydrogen radical with a substituent of a specified structure. When multiple positions in a particular structure are substituted by multiple specific substituents, each position of the substituent can be the same or different. The term "substitution" as used herein includes all permissible organic group substitutions. In a broad sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic organic groups. As described herein, heteroatomic nitrogen may be supplemented with a hydrogen substituent or any permissible organic group described above to complete its valence state. Furthermore, the present invention is not intended to limit permissible substituted organic groups in any way. The present invention considers the combination of substituents and variable groups to be beneficial in the treatment of diseases in the form of stable compounds. The term "stable" here means having a stable compound that, when tested over a sufficiently long period, maintains sufficient integrity of the compound structure, preferably remaining effective for a sufficiently long period, and is used herein for the purposes described above.

[0060] Pharmaceutical Compositions and Administration

[0061] Because the compounds of this invention can kill both Mycobacterium tuberculosis and non-tuberculous mycobacteria, and are used to treat diseases such as pulmonary tuberculosis, renal tuberculosis, bladder tuberculosis, and lung infections, the compounds of this invention, their stereoisomers, their optical isomers, their pharmaceutically acceptable salts, their crystal forms, their isotopic derivatives, their prodrugs, their metabolites, their solvates or hydrates thereof, and pharmaceutical compositions containing the compounds of this invention as the main active ingredient, can be used to prevent and / or treat (stabilize, alleviate or cure) diseases related to Mycobacterium tuberculosis and / or non-tuberculous mycobacteria (pulmonary tuberculosis, renal tuberculosis, bladder tuberculosis, lung infections, etc.).

[0062] The pharmaceutical compositions of the present invention comprise the compound of the present invention within a safe and effective range and a pharmaceutically acceptable excipient or carrier. "Safe and effective range" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably, 10-200 mg of the compound of the present invention per dose. Preferably, "one dose" is one capsule or tablet.

[0063] "Pharmaceutically acceptable carriers" refer to one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with and with the compounds of the present invention without significantly reducing the efficacy of the compounds. Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as... Wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0064] There are no particular limitations on the administration of the compounds or pharmaceutical compositions of the present invention. Representative administration methods include, but are not limited to, oral administration and parenteral administration (intravenous, intramuscular, or subcutaneous).

[0065] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following components: (a) fillers or compatibilizers, such as starch, lactose, sucrose, glucose, mannitol, and silica; (b) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or cassava starch, alginate, certain complex silicates, and sodium carbonate; (e) slowing agents, such as paraffin; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. Buffers may also be included in capsules, tablets, and pills.

[0066] Solid dosage forms such as tablets, sugar pills, capsules, pellets, and granules can be prepared using coatings and shells, such as casings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compound from such compositions can be delayed in a portion of the digestive tract. Examples of encapsulating components that can be used are polymeric substances and waxes. If necessary, the active compound may also be formed into microcapsules with one or more of the excipients described above.

[0067] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0068] In addition to these inert diluents, the composition may also contain auxiliaries such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances.

[0069] In addition to the active ingredient, the suspension may contain suspending agents, such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0070] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0071] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalers. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants as needed.

[0072] The compounds of this invention can be administered alone or in combination with other pharmaceutically acceptable compounds.

[0073] When administered in combination, the pharmaceutical composition further comprises one or more (two, three, four, or more) other pharmaceutically acceptable compounds. One or more (two, three, four, or more) of these other pharmaceutically acceptable compounds may be used simultaneously, separately, or sequentially with the compounds of the present invention for the prevention and / or treatment of diseases associated with Mycobacterium tuberculosis.

[0074] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is administered to the mammal (such as a human) requiring treatment. The dosage at the time of administration is the pharmaceutically considered effective dose. For a person weighing 60 kg, the daily dose is typically 1-2000 mg, preferably 20-500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the scope of the skill of a skilled physician.

[0075] The main advantages of this invention are:

[0076] 1. This invention discovers a novel inhibitor of mycobacteria (including tuberculous mycobacteria and non-tuberculous mycobacteria), providing a theoretical basis for drug development based on the mycoic acid synthesis pathway.

[0077] 2. The compounds of the present invention have excellent bactericidal effects and are highly specific, with no cross-resistance with existing anti-tuberculosis drugs, providing a breakthrough direction for the treatment of drug-resistant tuberculosis.

[0078] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, were generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight.

[0079] Example 1: Screening of InhA Inhibitors

[0080] Using InhA, a key enzyme in the mycolic acid synthesis pathway of mycobacteria, as a target, a number of potential novel mycobacterial inhibitors were screened based on structural virtual screening from drugs approved by the US FDA for the treatment of other diseases and compounds undergoing preclinical research. The relevant compounds were then subjected to drug susceptibility testing in Mycobacterium tuberculosis (Mtb H37Rv), BCG, and Mycobacterium abscessus (Mab ATCC19977). The experimental results are shown in Table 1.

[0081] Table 1. Screening of InhA Inhibitors

[0082]

[0083] As shown in Table 1, among the InhA inhibitors selected in the virtual screening above, compounds 4# (X-376), 6#, and 14# (X-396) exhibited the lowest minimum inhibitory concentration (MIC) against Mycobacterium tuberculosis (H37Rv). 90 The concentrations were all 62.5 μM, which is close to the minimum inhibitory concentration range (0.1-10 μM) of first-line drugs for tuberculosis treatment.

[0084] In addition, compounds 4# (X-376) and 14# (X-396) also showed excellent inhibitory effects on BCG and Mycobacterium abscessus (ATCC19977).

[0085] Specific information about X-376 and X-396 is shown in Table 2.

[0086] Table 2. Structural information of compounds X-376 and X-396

[0087]

[0088] Example 2: X-376 and X-396 Specific Targeting Mycobacteria

[0089] To determine whether X-376 and X-396 specifically target mycobacteria, the inventors, based on Example 1, further determined the minimum inhibitory concentration (MIC) of compounds X-376 and X-396, which have good antibacterial effects against mycobacteria, in Salmonella Typhimurium, Enterobacter cloacae, Klebsiella pneumoniae, and Staphylococcus aureus. 90 The results are shown in Table 3.

[0090] Table 3. Compounds X-376 and X-396 showed no antibacterial activity against other common clinical pathogens.

[0091]

[0092] The results showed that compounds X-376 and X-396 had no inhibitory effect on Salmonella Typhimurium, Enterobacter cloacae, Klebsiella pneumoniae, and Staphylococcus aureus, but had significant inhibitory effects on Mycobacterium tuberculosis (M. tuberculosis H37Rv) and Mycobacterium abscessus (M. abscessus ATCC 19977), further confirming that compounds X-376 and X-396 can specifically target mycobacteria.

[0093] Example 3: X-376 and X-396 showed no cross-resistance with existing anti-tuberculosis drugs.

[0094] To analyze the cross-resistance of compounds X-376 and X-396 with existing anti-tuberculosis drugs, we determined the minimum inhibitory concentrations (MICs) of compounds X-376 and X-396 against clinical XDR strains (extensively drug-resistant strains, typically resistant to 6–8 anti-tuberculosis drugs, as described below). 90 The results are shown in Table 4.

[0095] Among them, XDR-1 is resistant to isoniazid, rifampin, streptomycin, ethambutol, ofloxacin, levofloxacin, rifabutin, pyrazinamide, and para-aminosalicylic acid; XDR-2 is resistant to isoniazid, rifampin, streptomycin, ethambutol, ofloxacin, levofloxacin, rifabutin, pyrazinamide, kanamycin, capreomycin, and amikacin; XDR-3 is resistant to isoniazid, rifampin, streptomycin, ethambutol, ofloxacin, levofloxacin, rifabutin, pyrazinamide, kanamycin, capreomycin, amikacin, and para-aminosalicylic acid.

[0096] Table 4. Compounds X-376 and X-396 showed no cross-resistance with existing anti-tuberculosis drugs.

[0097]

[0098] The results showed that compounds X-376 and X-396 had MICs against clinical XDR strains (XDR-1, XDR-2, XDR-3). 90 Consistent with the standard strain (H37Rv) and clinically susceptible strains (DS-1 and DS-1), compounds X-376 and X-396 showed good inhibitory effects on clinical XDR strains, suggesting that compounds X-376 and X-396 do not exhibit cross-resistance with existing anti-tuberculosis drugs.

[0099] Example 4: X-376 and X-396 exhibit in vitro bactericidal activity.

[0100] To determine whether compounds X-376 and X-396 possess in vitro bactericidal activity, i.e., whether they are bactericidal agents (reduced viable bacterial count) rather than bacteriostatic agents (virulent bacterial count remains essentially unchanged), the applicant tested the effects of compounds X-376, 6#, and X-396 on the viable bacterial count (CFU) of standard strains of Mycobacterium tuberculosis (H37Rv) and Mycobacterium abscessus (ATCC 19977) at concentrations of 2×MIC and 10×MIC. The results for Mycobacterium tuberculosis (H37Rv) and Mycobacterium abscessus (ATCC 19977) are as follows: Figure 1 and Figure 2 As shown.

[0101] Figure 1 The results showed that compound 6# failed to reduce the number of viable Mycobacterium tuberculosis at concentrations of 2×MIC and 10×MIC, while compounds X-376 and X-396 significantly reduced the number of viable bacteria at concentrations of 2×MIC and 10×MIC. This indicates that compound 6# has no bactericidal effect, while compounds X-376 and X-396 are bactericidal agents that can effectively eliminate Mycobacterium tuberculosis in vitro, and their bactericidal effect is superior to that of first-line anti-tuberculosis drugs rifampin (RIF) and isoniazid (INH).

[0102] Figure 2 The results showed that compound 6# failed to reduce the viable count of Mycobacterium abscessus at concentrations of 2×MIC and 10×MIC, while compounds X-376 and X-396 significantly reduced the viable count at concentrations of 2×MIC and 10×MIC. This indicates that compound 6# has no bactericidal effect, while compounds X-376 and X-396 are bactericidal agents that can effectively eliminate Mycobacterium abscessus in vitro, and their bactericidal effect is superior to that of commonly used anti-Mycobacterium abscessus drugs such as amikacin (AMK) and clarithromycin (CL).

[0103] Example 5: X-376 and X-396 have the ability to eliminate dormant bacteria in vitro.

[0104] Dormant bacteria are bacteria in a dormant state that can usually resist the bactericidal effects of drugs. They are an important pathogenic reason for the long course of tuberculosis treatment. The dormant bacteria used in this example are dormant Mycobacterium tuberculosis (H37Rv) and Mycobacterium abscessus (ATCC 19977).

[0105] To test whether compounds X-376 and X-396 could effectively eliminate dormant bacteria, the inventors further determined the effect of compounds X-376 and X-396 on the viable count (CFU) of dormant Mycobacterium tuberculosis (H37Rv) and Mycobacterium abscessus (ATCC 19977) bacteria at compound concentrations of 2×MIC and 10×MIC, i.e., their bactericidal ability. The results for dormant Mycobacterium tuberculosis (H37Rv) and dormant Mycobacterium abscessus (ATCC 19977) are as follows: Figure 3 and Figure 4 As shown.

[0106] Figure 3 The results showed that compound 6# failed to reduce the viable number of dormant Mycobacterium tuberculosis bacteria at concentrations of 2×MIC and 10×MIC, while compounds X-376 and X-396 significantly reduced the viable number of dormant bacteria at concentrations of 2×MIC and 10×MIC. This indicates that compounds X-376 and X-396 can effectively eliminate dormant bacteria in vitro, and their bactericidal effect is superior to that of INH and streptomycin (STR). This suggests that compounds X-376 and X-396 may have a significant role in shortening the course of treatment for diseases related to Mycobacterium tuberculosis infection.

[0107] Figure 4 The results showed that compounds 6# and X-376 failed to reduce the viable number of dormant Mycobacterium abscessus at concentrations of 2×MIC and 10×MIC, while compound X-396 significantly reduced the viable number of dormant bacteria at concentrations of 2×MIC and 10×MIC. This indicates that compound X-396 can effectively eliminate dormant Mycobacterium abscessus in vitro, and its bactericidal effect is superior to that of AMK and CL. This suggests that compound X-396 may have a significant role in shortening the course of treatment for diseases related to Mycobacterium abscessus infection.

[0108] Example 6: X-376 and X-396 exhibit synergistic or additive effects with first-line anti-tuberculosis drugs.

[0109] To investigate the effects of compounds X-376 and X-396 in combination with first-line anti-tuberculosis drugs RIF and INH, the inventors determined the minimum inhibitory concentration (MIC) of compounds X-376 and X-396 against the standard strain of Mycobacterium tuberculosis (H37Rv) using the checkerboard method. 90 The combined efficacy of the drugs was evaluated using the fractional inhibitory concentration index (FICI) method, and the results are shown in Table 5.

[0110] FIC A = MIC of drug A when used in combination / MIC of drug A when used alone

[0111] FIC B= MIC of drug B when used in combination / MIC of drug B when used alone

[0112] FICI = FIC A + FIC B

[0113] Among them, FICI ≤ 0.5 indicates that the combination of the two drugs significantly enhances the antibacterial effect, that is, synergistic effect;

[0114] 0.5 < FICI ≤ 1 indicates that the combined effect of the two drugs is equal to or slightly better than that of using the drugs alone, that is, additive effect;

[0115] 1 < FICI ≤ 4.0 indicates that there is no significant synergy or antagonism in the combination of the two drugs, that is, no relevant effect;

[0116] FICI > 4.0 indicates that the combination of the two drugs may weaken the antibacterial effect, that is, antagonistic effect.

[0117] Table 5. Compounds X-376 and X-396 have synergistic or additive effects with first-line anti-tuberculosis drugs

[0118]

[0119] The results show that compounds X-376 and X-396 have synergistic or additive effects with the first-line anti-tuberculosis drugs RIF and INH, suggesting that compounds X-376 and X-396 can be used as combined drugs with first-line anti-tuberculosis drugs clinically, which helps to improve the clinical efficacy and reduce the emergence of drug resistance.

[0120] All the documents mentioned in the present invention are cited herein as references, as if each document was cited individually as a reference. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims

1. The use of a compound of formula (I) and a pharmaceutically acceptable salt thereof, characterized in that, A pharmaceutical composition for preparing a medicament for treating and / or preventing a disease associated with Mycobacterium tuberculosis and / or non-tuberculous mycobacteria, wherein R1, R2, and R3are each independently selected from the group consisting of H, D, C 1-4 alkyl, C 1-4 deuteroalkyl.

2. Use according to claim 1, characterized in that, The compound is selected from the group consisting of: 。 3. Use according to claim 1, characterized in that, The disease associated with Mycobacterium tuberculosis is tuberculosis; the disease associated with non-tuberculous mycobacteria is non-tuberculous mycobacterial disease; The tuberculosis is selected from the group consisting of pulmonary tuberculosis, renal tuberculosis, bladder tuberculosis, reproductive system tuberculosis, intestinal tuberculosis, tuberculous peritonitis, bone and joint tuberculosis, tuberculous meningitis, lymph node tuberculosis, skin tuberculosis, and blood-borne tuberculosis. The non-tuberculous mycobacterial disease is selected from the group consisting of pulmonary infection, skin and soft tissue infection, and lymphadenitis.

4. The use according to claim 1, characterized in that, The compounds described reduce the viable count of Mycobacterium tuberculosis and / or non-tuberculous mycobacterial standard strains by >1.5 log in 9 days at 2x MIC concentration 10 .

5. Use according to claim 4, characterized in that, reducing the viable count of Mycobacterium tuberculosis and / or non-tuberculous Mycobacterium standard strains by > 2 log at 2x MIC concentration 10 .

6. Use according to claim 5, characterized in that, reducing the viable count of Mycobacterium tuberculosis and / or non-tuberculous Mycobacterium standard strains by > 2.5 log at 2x MIC concentration 10 .

7. The use according to claim 1, characterized in that, The compounds have no antibacterial activity against Salmonella, Enterobacter cloacae, Klebsiella pneumoniae and / or Staphylococcus aureus, their antibacterial MIC 90 > 1000 μm.

8. The use according to claim 1, characterized in that, The compound inhibits the activity of InhA.

9. The use according to claim 1, characterized in that, The pharmaceutical composition further comprises other anti-Mycobacterium tuberculosis drugs and / or other anti-non-tuberculous mycobacterial drugs.

10. Use according to claim 9, characterized in that, The other anti-Mycobacterium tuberculosis drug is selected from the group consisting of rifampicin, isoniazid, pyrazinamide, ethambutol, streptomycin, ethionamide, p-aminosalicylic acid, capreomycin, kanamycin, ofloxacin, amikacin, cycloserine, bedaquiline, delamanid, or a combination thereof. The other anti-non-tuberculous mycobacterial drug is selected from the group consisting of amikacin, clofazimine, azithromycin, clarithromycin, cefoxitin, moxifloxacin, ciprofloxacin, imipenem, meropenem, or a combination thereof.

11. Use according to claim 1, characterized in that, The FICI of the compound of formula (I) in combination with isoniazid is ≤1 and >0.5; The FICI of the compound of formula (I) in combination with rifampicin is ≤0.5; wherein the compound of formula (I) is .

12. The use according to claim 1, characterized in that, The FICI of the compound of formula (I) in combination with rifampicin or isoniazid is ≤0.5; wherein the compound of formula (I) is .

13. The use according to claim 1, characterized in that, The pharmaceutical composition further comprises a pharmaceutically acceptable carrier, adjuvant, or excipient.

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