New delhi metallo-beta-lactamase-1 inhibitors

Abstract

The present invention relates to the use of (-) -epicatechin gallate, its stereoisomers, its hydrates, its solvates, its pharmaceutically acceptable salts, its hydrates or solvates of the pharmaceutically acceptable salts thereof or its pharmaceutically acceptable prodrugs in the manufacture of a medicament for the prevention and / or treatment of bacterial infection or diseases caused by bacterial infection, or in the manufacture of a medicament as a New Delhi metallo-beta-lactamase-1 (NDM-1) inhibitor, or in the manufacture of a medicament for antibacterial.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to a New Delhi metallo-β-lactamase-1 (NDM-1) inhibitor, which can be used to prevent and / or treat infections caused by NDM-1-producing bacteria. This invention also relates to the use of the NDM-1 inhibitor in combination with a β-lactam antibiotic in the preparation of a medicament for the prevention and / or treatment of bacterial infections or diseases caused by bacterial infections. Furthermore, this invention relates to a combination medicament or pharmaceutical composition containing the NDM-1 inhibitor and at least one β-lactam antibiotic. Background Technology

[0002] Antibiotics are a vital weapon in humanity's fight against bacterial infections. However, with the discovery and market launch of each class of antibiotics, while overcoming significant clinical challenges, drug resistance has also emerged. Carbapenems were once considered the most effective drugs for treating serious bacterial infections (including infections caused by drug-resistant bacteria, such as Gram-negative bacteria producing extended-spectrum β-lactamases). However, due to their widespread use and abuse, carbapenem resistance has become a major public health problem threatening human health worldwide. In the World Health Organization's 2017 list of "priority pathogens" for antibiotic resistance, three classes of carbapenem-resistant pathogens (carbapenem-resistant Acinetobacter baumannii, carbapenem-resistant Pseudomonas aeruginosa, and carbapenem-resistant, extended-spectrum β-lactamase-producing Enterobacteriaceae) were listed as "extremely important."

[0003] The production and spread of carbapenemases are important mechanisms of carbapenem antibiotic resistance. According to the Ambler classification system, carbapenemases are distributed in groups A (e.g., KPC, SME), B (e.g., NDM, VIM), and D (e.g., OXA-48, OXA-181). Among these, group B metallo-β-lactamases (NDM) have attracted significant attention due to their broad-spectrum resistance and rapid spread. NDM metallo-β-lactamases are carbapenemases capable of hydrolyzing almost all β-lactam drugs. Twenty-eight NDM alleles (NDM-1 to NDM-28) have been detected worldwide, with NDM-1 remaining the most common detected type.

[0004] NDM-1 can spread rapidly among different bacteria via plasmids or other mobile genetic elements, increasing the difficulty of monitoring and preventing carbapenem resistance caused by NDM-1. NDM-1-carrying strains are often resistant to most clinically used antibiotics. In many reports, tigecycline and colistin are the only drugs effective against NDM-1-positive strains. Currently, treatment options and clinical protocols for patients infected with NDM-1-carrying strains (especially pediatric patients) are very limited.

[0005] Combination therapy of serine β-lactamase inhibitors with β-lactam antibiotics has achieved good clinical efficacy. Given that currently marketed serine β-lactamase inhibitors have no significant effect on NDM-1-producing strains, researchers are actively seeking highly effective inhibitors of NDM-1. As a B1 class metallo-β-lactamase, NDM-1 has a typical αβ / βα sandwich structure, with its active site located in a cavity defined by flexible rings (L3 and L10), containing two divalent zinc ions bridged by hydroxide ions: Zn1 (coordinated with amino acids His120, His122, and His189) and Zn2 (coordinated with amino acids Asp124, Cys208, and His250).

[0006] Currently, there are over 500 NDM-1 inhibitors reported in the literature. Based on their mechanisms of action, they can be roughly divided into three categories: The first category of inhibitors directly acts on zinc ions at the active site of NDM-1, exerting inhibitory activity by chelating or coordinating with zinc ions. Since zinc-dependent enzymes also play important biological functions in the human body, the selectivity and in vivo safety of this type of inhibitor need to be carefully considered. The second category of inhibitors acts on amino acid residues of NDM-1, blocking the normal function of key amino acids or spatially hindering the binding of NDM-1 to substrates. The third category of inhibitors can simultaneously target zinc ions at the active site of NDM-1 and catalyze key amino acids, exerting an inhibitory effect on NDM-1. This type of inhibitor is considered to be the most promising NDM-1 inhibitor.

[0007] Despite significant progress in NDM-1 research, apart from the combination therapy of the bicyclic borate inhibitor VNRX-5133 and cefepime entering phase III clinical trials, there are currently no clinically available NDM-1 inhibitors. Finding highly effective and low-toxicity NDM-1 inhibitors remains a crucial scientific challenge. Summary of the Invention

[0008] The inventors have unexpectedly discovered that (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug have inhibitory activity against New Delhi metallo-β-lactamase-1 (NDM-1). Therefore, (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug can serve as an inhibitor of New Delhi metallo-β-lactamase-1 (NDM-1) for the prevention and / or treatment of bacterial infections or diseases caused by bacterial infections, particularly infections caused by New Delhi metallo-β-lactamase-1 (NDM-1)-producing bacteria or bacteria resistant to β-lactam antibiotics, or diseases caused by said infections. This NDM-1 inhibitor can be used in combination with β-lactam antibiotics for antibacterial purposes, particularly against NDM-1-containing superbugs. This invention was made based on the above findings.

[0009] This invention relates to the use of (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug in the preparation of medicaments for the prevention and / or treatment of bacterial infections or diseases caused by bacterial infections.

[0010] The present invention also relates to the use of (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug in the preparation of a medicament as a New Delhi metallo-β-lactamase-1 (NDM-1) inhibitor.

[0011] The present invention also relates to the use of (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug in the preparation of a medicament for antibacterial purposes.

[0012] The present invention also relates to the use of pharmaceutical compositions in the preparation of medicaments for the prevention and / or treatment of bacterial infections or diseases caused by bacterial infections, wherein the pharmaceutical composition comprises (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate or its pharmaceutically acceptable prodrug, and a pharmaceutically acceptable carrier or excipient.

[0013] In some embodiments, the pharmaceutical composition of the present invention further comprises a β-lactam antibiotic, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug.

[0014] The present invention also relates to the use of the pharmaceutical composition in the preparation of a medicament for antibacterial purposes.

[0015] The present invention also relates to the use of the pharmaceutical composition in the preparation of a medicament as a New Delhi metallo-β-lactamase-1 (NDM-1) inhibitor.

[0016] The present invention also relates to the use of (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug, in combination with a β-lactam antibiotic, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug, in the preparation of a medicament for the prevention and / or treatment of bacterial infections or diseases caused by bacterial infections.

[0017] The present invention also relates to the use of (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug, in combination with a β-lactam antibiotic, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug, in the preparation of a medicament for antibacterial purposes.

[0018] The present invention also relates to a combination drug comprising a first active ingredient and a second active ingredient, wherein the first active ingredient is (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or a pharmaceutically acceptable prodrug thereof, and the second active ingredient is a β-lactam antibiotic, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or a pharmaceutically acceptable prodrug thereof.

[0019] In some embodiments, the combination drug of the present invention further contains a pharmaceutically acceptable carrier or excipient.

[0020] In some embodiments, the first active ingredient in the combined medicament of the present invention is present in an effective amount for preventing or treating bacterial infections or diseases caused by bacterial infections.

[0021] In some embodiments, the second active ingredient in the combined medicament of the present invention is present in an effective amount for preventing or treating bacterial infections or diseases caused by bacterial infections.

[0022] In some embodiments, the first and second active ingredients of the combined medicament of the present invention are in the same formulation unit. In some embodiments, the first and second active ingredients of the combined medicament of the present invention are in different formulation units.

[0023] In some embodiments, the first and second active ingredients in the combined drug of the present invention are administered simultaneously, separately, or sequentially.

[0024] The present invention also relates to a pharmaceutical composition comprising a first active ingredient and at least one second active ingredient, wherein the first active ingredient is (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, a pharmaceutically acceptable salt, a hydrate or solvate of a pharmaceutically acceptable salt, or a pharmaceutically acceptable prodrug thereof; and the second active ingredient is a β-lactam antibiotic, its stereoisomer, its hydrate, its solvate, a pharmaceutically acceptable salt, a hydrate or solvate of a pharmaceutically acceptable salt, or a pharmaceutically acceptable prodrug thereof.

[0025] Optionally, the pharmaceutical composition further contains a pharmaceutically acceptable carrier or excipient.

[0026] In some embodiments, the first active ingredient in the pharmaceutical composition of the present invention is present in an effective amount for preventing or treating bacterial infections or diseases caused by bacterial infections.

[0027] In some embodiments, the second active ingredient in the pharmaceutical composition of the present invention is present in an effective amount for preventing or treating bacterial infections or diseases caused by bacterial infections.

[0028] The present invention also relates to (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate or its pharmaceutically acceptable prodrug, for the prevention and / or treatment of bacterial infections or diseases caused by bacterial infections.

[0029] The present invention also relates to (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate or its pharmaceutically acceptable prodrug, for antibacterial purposes.

[0030] The present invention also relates to (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate or its pharmaceutically acceptable prodrug, as an inhibitor of New Delhi metallo-β-lactamase-1 (NDM-1).

[0031] The present invention also relates to a method for preventing and / or treating bacterial infections or diseases caused by bacterial infections, comprising administering to a subject in need a preventive or therapeutically effective amount of (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug.

[0032] The present invention also relates to a method for preventing and / or treating bacterial infections or diseases caused by bacterial infections, comprising administering to a subject in need a preventative or therapeutically effective amount of (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug, and at least one preventative or therapeutically effective amount of a β-lactam antibiotic, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug.

[0033] The present invention also relates to another method of preventing and / or treating bacterial infections or diseases caused by bacterial infections, comprising administering to a subject in need a preventive or therapeutically effective amount of the combination drug or drug composition of the present invention.

[0034] In some embodiments, the bacterial infection described in this invention is an infection caused by New Delhi metallo-β-lactamase-1 (NDM-1)-producing bacteria or bacteria resistant to β-lactam antibiotics.

[0035] In some embodiments, the antibacterial activity described in this invention is bactericidal or bacteriostatic.

[0036] In some embodiments, the antibacterial agent of the present invention is an antibacterial agent against New Delhi metallo-β-lactamase-1 (NDM-1) bacteria or against bacteria resistant to β-lactam antibiotics.

[0037] In some embodiments, the bacteria producing New Delhi metallo-β-lactamase-1 (NDM-1) according to the present invention are Gram-negative bacteria that produce New Delhi metallo-β-lactamase-1 (NDM-1) (e.g., Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, Acinetobacter baumannii, or Citrobacter, etc.).

[0038] In some embodiments, the bacteria resistant to β-lactam antibiotics described in this invention are Gram-negative bacteria resistant to β-lactam antibiotics (e.g., Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, Acinetobacter baumannii, Citrobacter, etc.).

[0039] In some embodiments, the β-lactam antibiotics of the present invention are selected from: penicillins (such as: penicillin G, penicillin V, methicillin, oxacillin, cloxacillin, dicloxacillin, ampicillin, mecillin, temoxicillin, oxacillin, dicloxacillin, flucloxacillin, amoxicillin, piamocillin, carbenicillin, sulbenzylbenzyl, furazolidone, azoxystrobin, ticarcillin, piperacillin, etc.), cephalosporins (cefazoline, cefradine, cephalexin, cefadroxil, cefuroxime, cefotiam, cefaclor, cefuroxime axetil, cefprozil, cefotaxime, ceftriaxone, ceftazidime, cefoperazone, cefixime, cefpodoxime proxetil, cefepime, cefoperazone ... Cefuroxime, cefotaxime, cefotaxime, cefotaxime, cefomandole, cefepime, etc.), cephalosporins (cefoxitin, cefmetazole, cefminox, etc.), carbapenems (meropenem, imipenem, panipenem, ertapenem, faropenem, biapenem, doripenem, ipapenem, etc.), thiomycins, monocyclic β-lactams (aztreonam, carrumonam, etc.), and oxycephalosporins (lamiocefalexin, fluoxetine, etc.).

[0040] The various terms and phrases used in this invention have their general meanings known to those skilled in the art. In the event of any inconsistency between the terms and phrases mentioned and their known meanings, the meanings expressed in this invention shall prevail.

[0041] The term "(-)-Epicatechin gallate" used in this invention has the English name (-)-Epicatechin gallate (ECG), CAS number 1257-08-5, and molecular formula C2. 22 H 18 O 10 The structural formula is:

[0042]

[0043] The term “NDM-1” used in this invention is New Delhi metallo-β-lactamase-1.

[0044] The term "β-lactam antibiotics" as used in this invention refers to a large class of antibiotics having a β-lactam ring in their chemical structure. This includes the most commonly used penicillins and cephalosporins, as well as newly developed atypical β-lactam antibiotics such as cephalosporins, carbapenems, thiomycins, monocyclic β-lactams, and oxocephalosporins. Specific β-lactam antibiotics include, but are not limited to:

[0045] Penicillins include: penicillin G, penicillin V, methicillin, oxacillin, cloxacillin, dicloxacillin, ampicillin, mecillin, temoxicillin, oxacillin, dicloxacillin, flucloxacillin, amoxicillin, piamocillin, carbenicillin, sulfobenzylcillin, furazolidone, azlocillin, ticarcillin, piperacillin, etc.

[0046] Cephalosporins include: cefazolin, cefadroxil, cefalexin, cefadroxil, cefuroxime, cefotiam, cefaclor, cefuroxime axetil, cefprozil, cefotaxime, ceftriaxone, ceftazidime, cefoperazone, cefixime, cefpodoxime proxetil, cefepime, cefuroxime proxetil, cefotorol, cefotaxime, cefazolin, cefpirome, cefamandole, cefepime, etc.

[0047] Cephalosporins, such as cefoxitin, cefmetazole, and cefminox,

[0048] Monocyclic β-lactams, such as aztreonam and carrumonam,

[0049] Carbapenems include: meropenem, imipenem, panipenem, ertapenem, faropenem, biapenem, doripenem, ipapenem, etc.

[0050] Oxycephalosporins, such as latamoxef and fluoxetine.

[0051] The term "subject" as used in this invention includes mammals and humans, preferably humans.

[0052] The term "pharmaceutically acceptable salt" as used in this invention means a salt of the compound of this invention that is pharmaceutically acceptable and has the desired pharmacological activity of the parent compound. These salts include: salts formed by the addition of inorganic acids or acids formed with organic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; salts formed by the addition of organic acids, such as acetic acid, propionic acid, hexanoic acid, cyclopentadienoic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid, camphorsulfonic acid, glucoheponic acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, mucoaceric acid, etc.; or salts formed when acidic protons present on the parent compound are replaced by metal ions, such as alkali metal ions or alkaline earth metal ions; or coordination compounds formed with organic bases, such as ethanolamine, diethanolamine, triethanolamine, N-methylglucosamine, etc.

[0053] The pharmaceutical composition of this invention contains (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug, and a pharmaceutically acceptable carrier or excipient. The carrier includes, but is not limited to: ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphates, glycerol, sorbic acid, potassium sorbate, a mixture of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylate, beeswax, and lanolin. The excipient refers to an additive in the pharmaceutical preparation other than the active pharmaceutical ingredient. Its properties are stable, it has no incompatibilities with the main drug, does not produce side effects, does not affect efficacy, is not easily deformed, cracked, moldy, or infested by insects at room temperature, is harmless to the human body, has no physiological effects, does not produce chemical or physical reactions with the main drug, and does not affect the content determination of the main drug. Examples of excipients include binders, fillers, disintegrants, and lubricants in tablets; wine, vinegar, and medicinal juice in traditional Chinese medicine pills; the base portion in semi-solid preparations such as ointments and creams; and preservatives, antioxidants, flavoring agents, fragrances, solubilizers, emulsifiers, solvents, osmotic pressure regulators, and colorants in liquid preparations.

[0054] The pharmaceutical compositions or combinations thereof described in this invention can be administered by methods known in the art, such as, but not limited to, any of the following: oral, spray inhalation, rectal, nasal, buccal, topical, parenteral, such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal, and intracranial injection or infusion, or administration via an external implantation device. Oral, intraperitoneal, or intravenous administration is preferred.

[0055] As used in this invention, the term "effective amount" means an amount sufficient to achieve, or at least partially achieve, the desired effect. For example, a preventive effective amount means an amount sufficient to prevent, stop, or delay the onset of a disease; a therapeutic effective amount means an amount sufficient to cure or at least partially stop the disease and its complications in a patient already suffering from the disease. Determining such an effective amount is entirely within the capabilities of those skilled in the art. For example, an effective amount for therapeutic purposes will depend on the severity of the disease to be treated, the overall state of the patient's own immune system, the patient's general characteristics such as age, weight, and sex, the method of administration of the drug, and other concurrent treatments, etc.

[0056] The amount of (-)-epicatechin gallate and / or β-lactam antibiotics administered to a subject depends on the type and severity of the disease or condition, as well as the subject's characteristics such as general health, age, sex, weight, and tolerance to the drug. It also depends on the type of formulation and the route of administration, and factors such as the dosing cycle or time interval. Those skilled in the art can determine the appropriate dosage based on these and other factors. Generally, the daily dose of (-)-epicatechin gallate for treatment can be about 1 to 1000 mg, and the daily dose of β-lactam antibiotics for treatment can be about 1 to 1000 mg, which can be administered once or in multiple doses as appropriate. The (-)-epicatechin gallate and / or β-lactam antibiotics of the present invention can be provided in dosage units, with a content in the dosage unit ranging from 0.1 to 200 mg, for example, 1 to 100 mg.

[0057] In this invention, the term "combined drug" refers to a drug composition comprising (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug, and a β-lactam antibiotic, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug. This combined drug composition may be a pharmaceutical composition consisting of two active ingredients: (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug, and a β-lactam antibiotic, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug. It can also be a combination of two pharmaceutical ingredients, each consisting of (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate or its pharmaceutically acceptable prodrug, and a β-lactam antibiotic, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate or its pharmaceutically acceptable prodrug. In this combination therapy, the (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug, and the β-lactam antibiotic, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug, can be administered simultaneously or separately to the individual requiring treatment; or (e)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug can be administered first, followed by the β-lactam antibiotic, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug, after a certain interval; or (e)-lactam antibiotic, Its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate or its pharmaceutically acceptable prodrug, after a certain time interval, are administered (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate or its pharmaceutically acceptable prodrug.

[0058] In this invention, the term "pharmaceuticalally acceptable prodrug" refers to compounds that, upon administration to a subject, are typically hydrolyzed in the digestive tract, liver, or plasma to release (-)-epicatechin gallate or its metabolites in vivo. Typical prodrugs are pharmaceutically acceptable ethers or esters with a hydroxyl functional group. Pharmaceutically acceptable esters include alkyl esters (including acetates, butyrates, tert-butyrates, octadecyl esters, and neopentyl esters), phosphate esters, and sulfonates (i.e., esters derived from RSO2OH, where R is a lower alkyl or aryl group). Pharmaceutically acceptable ethers include lower alkyl ethers, such as C... 1-6 Alkyl ethers.

[0059] Beneficial technical effects of the present invention

[0060] The (-)-epicatechin gallate, its stereoisomer, its hydrate, its solvate, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt hydrate or solvate, or its pharmaceutically acceptable prodrug, as described in this invention, have inhibitory activity against New Delhi metallo-β-lactamase-1 (NDM-1) and can be used as a New Delhi metallo-β-lactamase-1 (NDM-1) inhibitor for the prevention and / or treatment of bacterial infections or diseases caused by bacterial infections, particularly infections caused by New Delhi metallo-β-lactamase-1 (NDM-1)-producing bacteria or bacteria resistant to β-lactam antibiotics. This NDM-1 inhibitor can be used in combination with β-lactam antibiotics for antibacterial purposes, particularly against NDM-1-producing superbugs. Attached Figure Description

[0061] Figure 1 The curves showing the changes in NDM-1 enzyme activity under different concentrations of (-)-epicatechin gallate are presented.

[0062] Figure 2 The curves showing the relationship between the reaction rate and the concentration of NDM-1 enzyme at different concentrations of the inhibitor (-)-epicatechin gallate are presented.

[0063] Figure 3 Lineweaver-Burke curves were shown at different concentrations of (-)-epicatechin gallate.

[0064] Figure 4 The fluorescence curves of NDM-1 enzyme under different concentrations of (-)-epicatechin gallate are shown.

[0065] Figure 5 The SPR patterns of (-)-epicatechin gallate binding to NDM-1 enzyme at different concentrations are shown.

[0066] Figure 6 It was shown that (-)-epicatechin gallate could restore the antibacterial activity of meropenem and biapenem against NDM-1-producing recombinant engineered bacteria in a dose-dependent manner;

[0067] Figure 7 The effect of Zn ions on the inhibition of NDM-1 enzyme activity by epicatechin gallate was shown. Detailed Implementation

[0068] The embodiments of the present invention will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of the invention. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer are followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0069] The experimental materials, reagents, and instruments involved in the embodiments of the present invention include:

[0070] (-)-Epicatechin gallate, purchased from Beijing Bailingwei Technology Co., Ltd., product number 426323;

[0071] 96-well UV analysis plate, purchased from CORNING, item number 3635;

[0072] 96-well cell culture plates, purchased from Costar, catalog number 3599;

[0073] The NDM-1 enzyme, purified from E. coli BL21(DE3) / pET30a(+)-blaNDM-1-M (an engineered bacterium expressing the mature NDM-1 protein gene), was provided by Professor You Xuefu's laboratory at the Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences.

[0074] HEPES FREE ACID, purchased from aMRESCO;

[0075] Meropenem trihydrate was purchased from TCI (Shanghai) Chemical Industry Development Co., Ltd.

[0076] The microplate reader, model Enspire 2300 Multiabel Reader, was purchased from PerkinElmer.

[0077] 96-hole blackboard, purchased from Costar, 3915;

[0078] EDC / NHS, purchased from Aladdin;

[0079] CM5 chip, purchased from Reichert;

[0080] Sodium acetate solution, purchased from GE Healthcare Bio-Sciences AB;

[0081] Ethanolamine, purchased from Sigma;

[0082] DMSO, purchased from aMRESCO;

[0083] PBST, purchased from Thermo Scientific, item number 28352;

[0084] PBS, purchased from Solarbio, catalog number P1010;

[0085] SPR, purchased from Reichert, model number 2SPR;

[0086] Piperacillin, purchased from Shanghai Maclean Biochemical Technology Co., Ltd.

[0087] Cefoperazone, cefotaxime, meropenem (MEPM), and biapenem were purchased from the National Institutes for Food and Drug Control.

[0088] E. coli BL21(DE3) / pET30a(+)-blaNDM-1 (an engineered bacterium expressing the full-length NDM-1 protein gene) was provided by Professor You Xuefu's laboratory at the Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences.

[0089] EDTA, purchased from Sangon Biotech (Shanghai) Co., Ltd.

[0090] Sealing film, purchased from Bemis;

[0091] ZnCl2, purchased from Beijing Bailingwei Technology Co., Ltd., product number 975486;

[0092] CCK-8 test kit, purchased from Bimake;

[0093] African green monkey kidney cells (Vero cells) were donated by Professor Li Yuhuan's laboratory at the Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences.

[0094] The HepG2 human liver cancer cells were donated by Professor He Qiyang's laboratory at the Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences.

[0095] HEPES buffer (10mM) is prepared as follows: Dissolve 2.383g of HEPES free active in 1000mL of distilled water, adjust the pH to 7.5 with NaOH, filter through a 0.45μm filter, and store at 0℃.

[0096] The LB liquid culture medium is prepared as follows: 5 g of yeast extract (OXOID, LP0021), 10 g of tryptone (OXOID, LP0042), 10 g of NaCl (Sinopharm Chemical Reagent Co., Ltd., 10019318), add distilled water to a final volume of 1000 mL, mix and sterilize at 121℃ for 15 min;

[0097] Vero medium for African green monkey kidney cells is prepared as follows: 450 mL MEM (Invitrogen) and 50 mL FBS (Gibco), mixed well and stored at 4°C.

[0098] The culture medium for human hepatocellular carcinoma HepG2 cells is prepared as follows: 450 mL of DMEM (Invitrogen) and 50 mL of FBS (Gibco), mixed well and stored at 4°C.

[0099] The meanings of the English abbreviations involved in the embodiments of the present invention are as follows:

[0100] NDM-1: New Delhi metallo-β-lactamase-1 (NDM-1);

[0101] HEPES: Hydroxyethyl piperazine ethanesulfonic acid;

[0102] EDC / NHS: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride / N-hydroxysuccinimide;

[0103] DMSO: dimethyl sulfoxide;

[0104] PBST: Phosphate-Buffered Saline with Tween TM -20);

[0105] PBSTD: Phosphate Tween buffer containing 1.25 v / v DMSO;

[0106] PBS: Phosphate buffer solution;

[0107] SPR: Surface plasmon resonance;

[0108] CFU: Colony-Forming Unit;

[0109] EDTA: ethylenediaminetetraacetic acid;

[0110] MIC: Minimum inhibitory concentration;

[0111] ECG: (-)-epicatechin gallate;

[0112] LB: LB medium (Luria-Bertani medium);

[0113] ZnCl2: Zinc chloride;

[0114] CCK-8: Cell Counting Kit-8;

[0115] OD: Optical density;

[0116] CC 50 : Half-maximal cytotoxicity (50%);

[0117] IC 50 : Half-maximal inhibitory concentration (50% inhibiting concentration).

[0118] In the following experiments, the concentration unit "M" represents mol / L, mM represents mmol / L, and μM represents μmol / L.

[0119] Example 1 (-) - Half-maximal inhibitory concentration (IC50) of epicatechin gallate against NDM-1 50 Measurement

[0120] (1) The final concentrations of (-)-epicatechin gallate were set to 50 μg / mL, 25 μg / mL, 12.5 μg / mL, 6.25 μg / mL, 3.125 μg / mL and 1.5625 μg / mL; 2 μl of (-)-epicatechin gallate diluted with distilled water according to the corresponding concentration was added to a 96-well UV plate, with five replicates for each group;

[0121] (2) The final concentration of NDM-1 enzyme was set to 0.2 μg / mL. 100 μl of enzyme solution diluted with 10 mM HEPES buffer was added to each well and incubated with (-)-epicatechin gallate at 37°C for 15 min.

[0122] (3) The final concentration of meropenem was set to 50 μM. 98 μl of meropenem solution diluted with 10 mM HEPES buffer was added to each well.

[0123] (4) Immediately place the 96-well UV plate in a microplate reader and measure the absorbance of the system at 300 nm every 1 min. Continuously measure at 37℃ for 15 min, calculate the enzyme activity at each concentration of (-)-epicatechin gallate, and analyze and calculate the IC50 using GraphPad Prism 5 software. 50 .

[0124] The formula for calculating enzyme activity is:

[0125] Enzyme activity (%) = (Reaction rate of NDM-1 enzyme inhibitor / Reaction rate of control group) × 100%

[0126] The formula for calculating the reaction rate is:

[0127]

[0128] T1 and T2 are the detection times, respectively.

[0129] The NDM-1 enzyme activity change curves under different concentrations of (-)-epicatechin gallate are shown in the figure below. Figure 1 As shown in the figure. The results showed that (-)-epicatechin gallate could effectively inhibit NDM-1 enzyme activity, and (-)-epicatechin gallate inhibited the IC50 of NDM-1. 50 The value is 4.48 μM.

[0130] Example 2: Kinetic study of the inhibitor (-)-epicatechin gallate against NDM-1.

[0131] 2.1 Reversibility Detection

[0132] (1) The final concentrations of (-)-epicatechin gallate were set at six concentration gradients: 50 μg / mL, 25 μg / mL, 12.5 μg / mL, 6.25 μg / mL, 3.125 μg / mL and 1.5625 μg / mL. 2 μl of each of the (-)-epicatechin gallate diluted with distilled water according to the corresponding concentration was added to a 96-well UV analysis plate.

[0133] (2) For the same concentration of (-)-epicatechin gallate, set up 6 enzyme concentration gradients, add 100 μl of NDM-1 enzyme solution diluted with 10 mM HEPES buffer to each well, and incubate with (-)-epicatechin gallate at 37°C for 15 min.

[0134] (3) The final concentration of meropenem was set to 50 μM. 98 μl of meropenem solution diluted with 10 mM HEPES buffer was added to each well.

[0135] (4) Immediately place the 96-well UV analysis plate in an ELISA reader and measure the absorbance of the system at 300 nm every 1 min at 37°C.

[0136] (5) Calculate the reaction rate at each inhibitor concentration, and plot the relationship curves between the reaction rate and the NDM-1 enzyme concentration at different inhibitor concentrations (-)-epicatechin gallate, such as... Figure 2 As shown.

[0137] The formula for calculating the reaction rate is shown below:

[0138]

[0139] T1 and T2 are the detection times, respectively.

[0140] Experimental results:

[0141] like Figure 2 As shown, without the addition of an inhibitor, a straight line passing through the origin can be obtained; when a gradient concentration of (-)-epicatechin gallate is added to the enzyme activity assay system, a set of straight lines passing through the origin but with a decreasing slope as the inhibitor concentration increases are obtained, suggesting that (-)-epicatechin gallate has a reversible inhibitory effect on NDM-1.

[0142] 2.2 Detection of Inhibition Type

[0143] (1) The final concentrations of (-)-epicatechin gallate were set at six concentration gradients: 50 μg / mL, 25 μg / mL, 12.5 μg / mL, 6.25 μg / mL, 3.125 μg / mL and 1.5625 μg / mL. 2 μl of each of the (-)-epicatechin gallate diluted with distilled water according to the corresponding concentration was added to a 96-well UV analysis plate.

[0144] (2) Set the final concentration of NDM-1 enzyme to 0.2 μg / mL, add 100 μl of enzyme solution diluted with 10 mM HEPES buffer to each well, and incubate with (-)-epicatechin gallate at 37℃ for 15 min.

[0145] (3) For the same concentration of (-)-epicatechin gallate, set up 7 meropenem concentration gradients, and add 98 μl of meropenem solution diluted with 10 mM HEPES buffer to each well.

[0146] (4) Immediately place the 96-well plate in an ELISA reader and measure the absorbance of the system at 300 nm every 1 min at 37°C;

[0147] (5) Calculate the reaction rate of (-)-epicatechin gallate at each concentration, and plot the Lineweaver-Burke curves (e.g., 1 / [S] as the x-axis and 1 / V as the y-axis) for different concentrations of (-)-epicatechin gallate. Figure 3 As shown in the figure, the inhibitory effects of (-)-epicatechin gallate on NDM-1 were analyzed.

[0148] Experimental results:

[0149] The results are as follows Figure 3 As shown, Lineweaver-Burke curves were plotted at different concentrations of (-)-epicatechin gallate, resulting in a set of straight lines intersecting the negative half of the X-axis. The slope of the straight lines increased with increasing concentration of (-)-epicatechin gallate, indicating that (-)-epicatechin gallate inhibits the activity of NDM-1 in a non-competitive manner.

[0150] Example 3(-) - Detection of the interaction between epicatechin gallate and NDM-1

[0151] 3.1 Fluorescence Quenching Experiment

[0152] Fluorescence detection was performed on a 96-well blackboard with a detection volume of 200 μl and a detection temperature of 37 °C.

[0153] (1) Set the final concentration of NDM-1 enzyme solution to 647 μg / mL, and add 198 μl of enzyme solution to each well;

[0154] (2) Set up five concentration gradients for (-)-epicatechin gallate: 50 μg / mL, 25 μg / mL, 12.5 μg / mL, 6.25 μg / mL and 3.125 μg / mL; add 2 μl of (-)-epicatechin gallate diluted with distilled water to each well according to the corresponding concentration;

[0155] Two control groups were set up simultaneously. One control group was given only the inhibitor (-)-epicatechin gallate, without the NDM-1 enzyme; the other control group was given only the NDM-1 enzyme, without the inhibitor (-)-epicatechin gallate.

[0156] (3) Set the excitation wavelength to 270 nm, scan the emission light from 290 nm to 500 nm, and plot the fluorescence curves at different inhibitor concentrations (-)-epicatechin gallate, such as... Figure 4 As shown.

[0157] Experimental results:

[0158] The results showed that (-)-epicatechin gallate dose-dependently reduced the fluorescence intensity of NDM-1, while shifting the maximum emission wavelength of the NDM-1 enzyme from 347 nm (without (-)-epicatechin gallate) to 359 nm (with 50 μg / mL (-)-epicatechin gallate). These results suggest that (-)-epicatechin gallate may bind to the aromatic amino acids of the NDM-1 enzyme, causing a polarization of its microenvironment and thus altering the emission spectrum of NDM-1.

[0159] 3.2 Surface Plasmon Resonance (SPR) Experiment

[0160] (1) Inject the EDC / NHS mixture through channels 1 and 2 to activate the chip surface. Dilute the NDM-1 enzyme with sodium acetate solution of appropriate pH and fix it separately in channel 1, with channel 2 serving as a blank control. After the NDM-1 enzyme is coupled and immobilized, inject ethanolamine through channels 1 and 2 to seal the chip;

[0161] (2) A concentration gradient of (-)-epicatechin gallate was set up at 50 μg / mL, 25 μg / mL, 12.5 μg / mL, 6.25 μg / mL, 3.125 μg / mL, 1.5625 μg / mL, and 0.78125 μg / mL, and diluted with PBSTD (PBST buffer containing 1.25 v / v % DMSO). Different concentrations of (-)-epicatechin gallate were sequentially injected into the protein channel, with a PBSTD blank control set between every three samples. The binding of (-)-epicatechin gallate to NDM-1 enzyme at different concentrations was observed.

[0162] (3) The SPR binding plot of (-)-epicatechin gallate with NDM-1 enzyme was fitted using Trace Drawer software, and the affinity constant K was calculated. D ,like Figure 5 As shown.

[0163] Experimental results:

[0164] Figure 5 The SPR profiles of (-)-epicatechin gallate binding to NDM-1 enzyme at different concentrations are shown. The SPR results suggest a moderate-strength interaction between (-)-epicatechin gallate and NDM-1, K...D The value is 8.02e -5 M.

[0165] Example 4 (-) - Detection of the synergistic effect of epicatechin gallate on β-lactam antibiotics

[0166] 4.1(-)-Epicatechin gallate can restore the antibacterial activity of various β-lactam antibiotics against NDM-1-resistant strains.

[0167] To investigate whether (-)-epicatechin gallate has inhibitory activity against NDM-1 enzyme at the bacterial cell level, an in vitro drug susceptibility test was performed using NDM-1 recombinant engineered bacteria to determine the minimum inhibitory concentration (MIC) of β-lactam antibiotics against NDM-1 recombinant engineered bacteria and empty plasmid control bacteria.

[0168] (1) In sterile 96-well cell culture plates, 100 μl of gradient concentrations of β-lactam antibiotic solutions were added (cefotaxime and cefotaxime were set at a concentration gradient of 256.0 μg / mL to 0.125 μg / mL; piperacillin was set at a concentration gradient of 1024.0 μg / mL to 0.5 μg / mL; meropenem and biapenem were set at a concentration gradient of 64 μg / mL to 0.03125 μg / mL). The concentration gradients of the above antibiotics were respectively prepared using a solution containing approximately 4 × 10⁻⁶ μg / mL of β-lactam antibiotics. 5 It was prepared by diluting LB medium containing CFU / mL NDM-1 producing bacteria;

[0169] (2) The concentration of the inhibitor (-)-epicatechin gallate was set at 32 μg / mL, the concentration of the positive control drug EDTA was set at 32 μg / mL, and the solvent DMSO control well and the negative growth control well (with sterile culture medium added) were also set.

[0170] (3) Seal the 96-well cell culture plate with sealing film and incubate it in a 37°C incubator in the dark.

[0171] (4) After culturing for about 20 hours, observe and record the bacterial growth of each experimental group, and calculate the minimum inhibitory concentration (MIC) of β-lactam antibiotics against NDM-1 recombinant engineered bacteria and empty plasmid control bacteria.

[0172] Experimental results:

[0173] The results are shown in Table 1. 32 μg / mL (-)-epicatechin gallate reduced the MICs of piperacillin, cefoperazone, meropenem, and biapenem against NDM-1 recombinant engineered bacteria to 1 / 2, 1 / 4, 1 / 4, and 1 / 4 of their original values, respectively.

[0174] Table 1 (-) - Antibacterial effects of epicatechin gallate on NDM-1 recombinant engineered bacteria and empty plasmid control bacteria

[0175]

[0176] 4.2(-)-Epicatechin gallate can dose-dependently restore the antibacterial activity of carbapenem antibiotics against NDM-1-resistant strains.

[0177] (1) Add 100 μl of carbapenem antibiotic solutions (meropenem / biapenem, with a concentration gradient of 64 μg / mL to 0.0625 μg / mL) to a sterile 96-well cell culture plate. The above gradients are respectively treated with solutions containing approximately 4 × 10⁻⁶ μg / mL of carbapenem antibiotics. 5 It was prepared by diluting LB medium containing CFU / ml NDM-1 producing bacteria;

[0178] (2) The combined effect of (-)-epicatechin gallate and carbapenem antibiotics was determined by the checkerboard method. The concentration of the inhibitor (-)-epicatechin gallate was set at 128 μg / mL to 16 μg / mL. Two control groups were set up at the same time. One control group was given antibiotics alone without (-)-epicatechin gallate. The other control group was given (-)-epicatechin gallate alone without antibiotics.

[0179] (3) Seal the 96-well cell culture plate with sealing film and incubate it in a 37°C incubator in the dark.

[0180] (4) After culturing for about 20 hours, observe and record the bacterial growth of each experimental group, and calculate the minimum inhibitory concentration (MIC) of the antibiotics against the NDM-1 recombinant engineered bacteria.

[0181] Experimental results:

[0182] The results are as follows Figure 6 As shown, (-)-epicatechin gallate itself has no significant antibacterial activity at concentrations of 16 μg / mL to 128 μg / mL, but it can restore the antibacterial activity of meropenem and biapenem against NDM-1-producing recombinant engineered bacteria in a dose-dependent manner. When 64 μg / mL of (-)-epicatechin gallate is added, the MICs of meropenem and biapenem are reduced to 1 / 16 and 1 / 8 of their original values, respectively.

[0183] Example 5: Zinc Ion Replenishment Experiment

[0184] (1) The detection system consisted of 200 μl of NDM-1-producing recombinant engineered bacteria, with an inoculum size of approximately 2 × 10⁻⁶. 5 CFU / mL;

[0185] (2) Set the final concentration of meropenem and (-)-epicatechin gallate to 16 μg / mL, and add ZnCl2 in an equimolar amount of (-)-epicatechin gallate; the positive control drug EDTA was treated in the same way.

[0186] (3) Seal the 96-well cell culture plate with sealing film and incubate it in a 37°C incubator in the dark.

[0187] (4) After culturing for about 30 hours, observe and record the bacterial growth of each experimental group, and use an enzyme-linked immunosorbent assay (ELISA) reader to measure the absorbance of the system at 600 nm.

[0188] Experimental results:

[0189] The results are as follows Figure 7 As shown, 16 μg / mL (-)-epicatechin gallate restored the antibacterial activity of meropenem and inhibited the growth of NDM-1-resistant bacteria; however, the synergistic effect of (-)-epicatechin gallate on meropenem was almost completely reversed by exogenous zinc ions; the positive control drug EDTA showed a similar trend. This indicates that the NDM-1 inhibitory activity of (-)-epicatechin gallate is related to its effect on zinc ions.

[0190] Example 6 Cytotoxicity evaluation of NDM-1 inhibitor (-)-epicatechin gallate

[0191] (1) The concentration of Vero cells in the logarithmic growth phase of African green monkey kidney cells was adjusted to approximately 2 × 10⁻⁶ using fresh culture medium. 4 The concentration of HepG2 human liver cancer cells was adjusted to approximately 5 × 10⁶ cells / mL. 4 Cells / mL. Seed 100 μL of cells into sterile 96-well cell culture plates and incubate at 37°C in a 5% CO2 cell culture incubator.

[0192] (2) After culturing for 24 hours and the cells adhered to the cell wall, 100 μL of serum-free medium diluted with (-)-epicatechin gallate was added to each well to make the final concentration gradients 200, 100, 50, 25, 12.5, 6.25 and 3.125 μg / mL. A DMSO solvent control group, a cell control group and a background (medium) control group were set up. The cells were cultured for another 48 hours in a 37℃, 5% CO2 cell incubator.

[0193] (3) Remove the 96-well plate, discard the culture medium, add 100 μL of CCK-8 detection reagent, and continue incubation for 1-2 hours until the OD of the cell control group is reached. 450 The solution should reach approximately 1. Place it on a horizontal shaker and shake at low speed for 10 minutes to ensure it is thoroughly mixed.

[0194] (4) The absorbance at 450 nm was measured using an ELISA reader. The cell viability (%) was calculated using the formula: Cell viability (%) = (OD of drug-treated group - Background OD) / (OD of control group - Background OD) × 100%. The CC was calculated using GraphPad Prism. 50 .

[0195] Experimental results:

[0196] The results are shown in Table 2. (-)-Epicatechin gallate exhibited relatively low cytotoxicity against CSA2 in Vero cells. 50 At 106.3 μM, it had an effect on CC in HepG2 cells. 50 It is 80.76 μM.

[0197] Table 2(-) - Effects of catechin gallate on cellular CC 50 value

[0198]

Claims

1. Use of a pharmaceutical composition in the preparation of a medicament for the prevention and / or treatment of bacterial infections or diseases caused by bacterial infections, wherein the pharmaceutical composition comprises an active ingredient and a pharmaceutically acceptable carrier, wherein the active ingredient comprises a first active ingredient and a second active ingredient, the first active ingredient being (-)-epicatechin gallate or a pharmaceutically acceptable salt thereof, and the second active ingredient being a β-lactam antibiotic or a pharmaceutically acceptable salt thereof, wherein the β-lactam antibiotic is selected from piperacillin, cefoperazone, meropenem, and biapenem, and the bacterial infection is an infection caused by New Delhi metallo-β-lactamase-1-producing Gram-negative bacteria or Gram-negative bacteria resistant to β-lactam antibiotics, wherein the New Delhi metallo-β-lactamase-1-producing Gram-negative bacteria or Gram-negative bacteria resistant to β-lactam antibiotics is selected from Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, Acinetobacter baumannii, and Citrobacter.

2. Use of a pharmaceutical composition in the preparation of a medicament for antibacterial purposes, wherein the pharmaceutical composition comprises an active ingredient and a pharmaceutically acceptable carrier, wherein the active ingredient comprises a first active ingredient and a second active ingredient, the first active ingredient being (-)-epicatechin gallate or a pharmaceutically acceptable salt thereof, and the second active ingredient being a β-lactam antibiotic or a pharmaceutically acceptable salt thereof, wherein the β-lactam antibiotic is selected from piperacillin, cefoperazone, meropenem, and biapenem, and the antibacterial activity is against New Delhi metallo-β-lactamase-1-producing Gram-negative bacteria or against Gram-negative bacteria resistant to β-lactam antibiotics, wherein the New Delhi metallo-β-lactamase-1-producing Gram-negative bacteria or Gram-negative bacteria resistant to β-lactam antibiotics are selected from Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, Acinetobacter baumannii, and Citrobacter.

3. Use of a pharmaceutical composition in the preparation of a medicament as a New Delhi metallo-β-lactamase-1 inhibitor, wherein the pharmaceutical composition comprises an active ingredient and a pharmaceutically acceptable carrier, wherein the active ingredient comprises a first active ingredient and a second active ingredient, the first active ingredient being (-)-epicatechin gallate or a pharmaceutically acceptable salt thereof, and the second active ingredient being a β-lactam antibiotic or a pharmaceutically acceptable salt thereof, said β-lactam antibiotic being selected from: piperacillin, cefoperazone, meropenem, and biapenem.

4. The use according to claim 2, wherein the antibacterial activity is bactericidal or bacteriostatic.

5. The use of (-)-epicatechin gallate or a pharmaceutically acceptable salt thereof, alone as an active ingredient, in combination with a β-lactam antibiotic or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for the prevention and / or treatment of bacterial infections or diseases caused by bacterial infections, wherein the bacterial infection is caused by New Delhi metallo-β-lactamase-1-producing Gram-negative bacteria or Gram-negative bacteria resistant to β-lactam antibiotics; wherein the β-lactam antibiotic is selected from: piperacillin, cefoperazone, meropenem, and biapenem; wherein the New Delhi metallo-β-lactamase-1-producing Gram-negative bacteria or Gram-negative bacteria resistant to β-lactam antibiotics is selected from Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, Acinetobacter baumannii, and Citrobacter.

6. The use of (-)-epicatechin gallate or a pharmaceutically acceptable salt thereof, alone as an active ingredient, in combination with a β-lactam antibiotic or a pharmaceutically acceptable salt thereof, in the preparation of an antibacterial medicament, wherein the antibacterial activity is against New Delhi metallo-β-lactamase-1-producing Gram-negative bacteria or against Gram-negative bacteria resistant to β-lactam antibiotics; wherein the β-lactam antibiotic is selected from: piperacillin, cefoperazone, meropenem, and biapenem; wherein the New Delhi metallo-β-lactamase-1-producing Gram-negative bacteria or Gram-negative bacteria resistant to β-lactam antibiotics is selected from Klebsiella pneumoniae, Escherichia coli, Enterobacter cloacae, Acinetobacter baumannii, and Citrobacter.

7. The use according to claim 6, wherein the antibacterial activity is bactericidal or bacteriostatic.

8. A combination drug, wherein the active ingredient comprises a first active ingredient and at least one second active ingredient, the first active ingredient being (-)-epicatechin gallate or a pharmaceutically acceptable salt thereof, and the second active ingredient being a β-lactam antibiotic or a pharmaceutically acceptable salt thereof, said β-lactam antibiotic being selected from piperacillin, cefoperazone, meropenem, and biapenem.

9. The combination drug of claim 8, further comprising a pharmaceutically acceptable carrier.

10. The combination medicament of claim 8, wherein the first active ingredient is present in an effective amount for preventing or treating bacterial infection or disease caused by bacterial infection; The second active ingredient is present in an effective amount to prevent or treat bacterial infections or diseases caused by bacterial infections.

11. The combination drug of claim 8, wherein the first active ingredient and the second active ingredient are in the same formulation unit, or the first active ingredient and the second active ingredient are in different formulation units.

12. The combination drug of claim 8, wherein the first active ingredient and the second active ingredient are administered simultaneously, separately, or sequentially.

13. A pharmaceutical composition comprising an active ingredient and a pharmaceutically acceptable carrier, wherein the active ingredient comprises a first active ingredient and a second active ingredient, the first active ingredient being (-)-epicatechin gallate or a pharmaceutically acceptable salt thereof, and the second active ingredient being a β-lactam antibiotic or a pharmaceutically acceptable salt thereof, said β-lactam antibiotic being selected from piperacillin, cefoperazone, meropenem, and biapenem.

14. The pharmaceutical composition of claim 13, wherein the first active ingredient is present in an effective amount for preventing or treating bacterial infection or disease caused by bacterial infection; The second active ingredient is present in an effective amount to prevent or treat bacterial infections or diseases caused by bacterial infections.

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