Antibacterial or antifungal composition and pharmaceutical composition for preventing or treating bacterial or fungal infections, comprising VLX600

Abstract

The present invention relates to an antibacterial or antifungal composition and a pharmaceutical composition for preventing or treating bacterial or fungal infections, both comprising VLX600. The antibacterial composition comprising VLX600 according to one aspect of the present invention exhibits antibacterial or antifungal activity against a wide range of strains, including Mycobacterium spp., Escherichia spp., Pseudomonas spp., Staphylococcus spp., Acinetobacter spp., Candida spp., and the like. The pharmaceutical composition comprising VLX600 demonstrates excellent antibacterial, antifungal, and therapeutic efficacy against pathogens and can be effectively utilized for the prevention or treatment of bacterial or fungal infections. Furthermore, VLX600 exhibits a synergistic effect with conventional antibiotics and thus is effective in treating infections caused by antibiotic-resistant bacterial strains.

Description

Antibacterial or antifungal composition comprising VLX600 and pharmaceutical composition for preventing or treating bacterial or fungal infections

[0001] The present invention relates to an antibacterial or antifungal composition comprising VLX600 and a pharmaceutical composition for preventing or treating bacterial or fungal infections.

[0002] Beginning with penicillin, the first antibiotic discovered in 1920, various types of antibiotics, including cephalosporins, quinolones, and tetracyclines, have been discovered and developed, and the use of these antibiotics has significantly reduced the mortality rate from bacterial infections.

[0003] However, along with the ever-increasing use of antibiotics, the incidence of antibiotic-resistant bacteria is rapidly increasing every year, while the pace of development of new antibiotics continues to decrease.

[0004] According to a WHO report, 700,000 people worldwide die each year due to antibiotic-resistant bacteria, and if this upward trend continues, the number is expected to increase to 10,000,000 by 2050, highlighting the need to develop new antibiotics to address this issue.

[0005] Among these, six strains of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterbacter spp., collectively known as ESKAPE, have been reported to have particularly high antibiotic resistance, and are frequently multidrug resistant (MDR), posing a significant public health problem.

[0006] In addition, Mycobacterium spp., a genus that includes Mycobacterium tuberculosis, the causative agent of tuberculosis, and nontuberculous mycobacteria (NTM) such as M. abscessus and M. intracellulare, have innate resistance to various antibiotics, and resistance is rapidly increasing even to highly susceptible antibiotics, making treatment difficult.

[0007] However, despite the steady increase in antibiotic-resistant bacteria, the pace at which new antibiotics are developed to counter them is decreasing significantly each year, making it urgent to revitalize the new antibiotic development market.

[0008] In addition, fungal infections, particularly those caused by Candida spp., are emerging as a serious clinical problem. Candidiasis causes opportunistic infections in immunocompromised patients, and invasive candidiasis has a particularly high mortality rate. Furthermore, the increasing number of strains resistant to existing antifungal agents necessitates the development of new antifungal agents.

[0009] Meanwhile, it generally takes about 15 years for a drug to be developed and approved for use, and it requires astronomical costs and labor.

[0010] Drug repositioning is a development strategy that uses drugs that have already been developed and approved for other purposes, or that have not yet been approved but are undergoing clinical trials, for a different purpose. It has the clear advantage of significantly reducing the time and cost required for development, as it uses drugs that have already been proven to be effective and safe to a certain extent by simply changing their purpose.

[0011] Accordingly, the inventors of this application sought to develop new antibiotic uses for existing drugs through drug repurposing. Through a large-scale screening of 3,200 clinical compounds, the inventors of this application identified new antibiotic and antifungal uses for VLX600.

[0012] VLX600 is a drug developed as an anticancer agent that induces cancer cell-specific apoptosis by inhibiting mitochondrial respiration by interfering with intracellular iron metabolism. A phase 1 clinical trial of VLX600 was conducted, and in the clinical trial, VLX600 exhibited generally acceptable levels of toxicity and adverse effects, and no dose-limiting toxicity was identified.

[0013] The inventors of the present application, through in-depth research, confirmed the growth inhibition effect of VLX600 on various bacteria and fungi, the elimination effect within infected cells, and the therapeutic effect in an animal model of infection, thereby deriving new antibacterial and antifungal uses of VLX600, and uses for the prevention and treatment of bacterial and fungal infections.

[0014] [Prior Art Literature]

[0015] [Patent Document]

[0016] (Patent Document 0001) International Patent Publication No. WO 2012 / 128689 A1

[0017] (Patent Document 0002) Korean Patent No. 10-1937279

[0018] [Non-patent literature]

[0019] (Non-patent Document 0001) Fryknas, M., Zhang, X., Bremberg, U. et al. Iron chelators target both proliferating and quiescent cancer cells. Sci Rep 6, 38343 (2016)

[0020] (Non-patent Document 0002) Mody, K., Mansfield, AS, Vemireddy, L. et al. A phase I study of the safety and tolerability of VLX600, an Iron Chelator, in patients with refractory advanced solid tumors. Invest New Drugs 37, 684-692 (2019).

[0021] One aspect of the present invention is to provide an antibacterial or antifungal composition comprising VLX600.

[0022] Another aspect provides the use of VLX600 for the preparation of an antibacterial or antifungal composition.

[0023] Another aspect is to provide a pharmaceutical composition for preventing or treating bacterial or fungal infections, comprising VLX600 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0024] Another aspect provides a method of preventing or treating a bacterial or fungal infection comprising administering VLX600 or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0025] Another aspect provides the use of VLX600 or a pharmaceutically acceptable salt thereof for use in the manufacture of a pharmaceutical composition for the prevention or treatment of a bacterial or fungal infection.

[0026] Another aspect provides a use of VLX600 or a pharmaceutically acceptable salt thereof for the prevention or treatment of a bacterial or fungal infection.

[0027] Another aspect is to provide a health functional food for preventing or improving bacterial or fungal infections containing VLX600.

[0028] Another aspect provides the use of VLX600 for use in the manufacture of a health functional food for the prevention or amelioration of bacterial or fungal infections.

[0029] Another aspect provides a use of VLX600 for preventing or ameliorating a bacterial or fungal infection.

[0030] Another aspect is to provide an antibacterial or antifungal over-the-counter drug product comprising VLX600.

[0031] Another aspect provides the use of VLX600 for the manufacture of antibacterial or antifungal over-the-counter drugs.

[0032] Another aspect is to provide an antibacterial or antifungal cosmetic composition comprising VLX600.

[0033] Another aspect provides the use of VLX600 for the preparation of an antibacterial or antifungal cosmetic composition.

[0034] Another aspect is to provide an antibacterial or antifungal feed composition comprising VLX600.

[0035] Another aspect provides the use of VLX600 for the preparation of an antibacterial or antifungal feed composition.

[0036] One aspect of the present invention provides an antibacterial or antifungal composition comprising VLX600.

[0037] Another aspect provides the use of VLX600 for the preparation of an antibacterial or antifungal composition.

[0038] VLX600 is an oxidative phosphorylation (OXPHOS) inhibitor and iron chelator.

[0039] The CAS number of the above VLX600 is 327031-55-0 and has the structure of a compound represented by the following chemical formula 1:

[0040] [Chemical Formula 1]

[0041]

[0042] In this specification, the term “antibacterial or antifungal” means the ability to resist bacteria, or any mechanism that is performed to resist or inhibit reproduction of microorganisms such as bacteria or fungi, or to defend against the action of microorganisms such as bacteria or fungi.

[0043] In one specific example, the antibacterial composition may have antibacterial activity against bacteria.

[0044] In one specific example, the antifungal composition may have antifungal activity against fungi.

[0045] In one specific example, the antibacterial or antifungal composition may have antibacterial or antifungal activity against one or more strains selected from the group consisting of Mycobacterium strains, Escherichia strains, Pseudomonas strains, Staphylococcus strains, Acinetobacter strains, and Candida strains.

[0046] Specifically, the antibacterial composition may have antibacterial activity against one or more strains selected from the group consisting of Mycobacterium strains, Escherichia strains, Pseudomonas strains, Staphylococcus strains, and Acinetobacter strains.

[0047] Specifically, the antifungal composition may have antifungal activity against strains of the genus Candida.

[0048] In one specific example, the strain of the genus Mycobacterium is Mycobacterium abscessus, Mycobacterium abscessus subsp.abscessus, Mycobacterium abscessus subsp.massiliense, Mycobacterium abscessus subsp.bolletii, Mycobacterium smegmatis, Mycobacterium intracellulare, Mycobacterium avium, Mycobacterium kansasii, Mycobacterium fortuitum, Mycobacterium Mycobacterium gordonae, Mycobacterium osloensis, Mycobacterium phlei, Mycobacterium terrae, Mycobacterium chelonae, Mycobacterium mucogenicum, Mycobacterium peregrinum, Mycobacterium simiae, Mycobacterium wolinskyi, Mycobacterium paragordonae, Mycobacterium ulcerans, Mycobacterium marinum, Mycobacterium bovis, Mycobacterium bovis BCG,It may be at least one selected from the group consisting of Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium canetti, Mycobacterium caprae, Mycobacterium microti, Mycobacterium tuberculosisK strain and Mycobacterium leprae.

[0049] The above Escherichia genus strain may be at least one selected from the group consisting of Escherichia coli, Escherichia albertii, Escherichia blattae, Escherichia fergusonii, Escherichia hermannii, and Escherichia vulneris.

[0050] The above Pseudomonas genus strain may be at least one selected from the group consisting of Pseudomonas aeruginosa, Pseudomonas mendocina, Pseudomonas stutzeri, Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas syringae, Pseudomonas alcaligenes, Pseudomonas luteola, and Pseudomonas oryzihabitans.

[0051] The above Staphylococcus strains include Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Staphylococcus lugdunensis, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus capitis, Staphylococcus warneri, Staphylococcus schleiferi, Staphylococcus intermedius, and Staphylococcus. It may be one or more selected from the group consisting of Staphylococcus pyogens.

[0052] The Acinetobacter genus strain may be at least one selected from the group consisting of Acinetobacter baumannii, Acinetobacter calcoaceticus, Acinetobacter lwoffii, Acinetobacter junii, Acinetobacter haemolyticus, Acinetobacter radioresiststens, Acinetobacter ursingii, Acinetobacter gerneri, and Acinetobacter nosocomialis.

[0053] The Candida genus strain may be at least one selected from the group consisting of Candida albicans, Candida glabrata, Candida tropicalis, Candida pseudotropicalis, Candida parapsilosis, Candida auris, Candida krusei, Candida glabrata, Candida dubliniensis, Candida lusitaniae, Candida kefyr, Candida haemulonii, and Candida rugosa.

[0054] In one specific example, the antibacterial or antifungal composition may additionally comprise an antibiotic.

[0055] The above antibiotics include clarithromycin, polymyxin B, erythromycin, isoniazid, rifampicin, ethambutol, SQ-109, pyrazinamide, streptomycin, gentamicin, kanamycin, apramycin, capreomycin, ethionamide, prothionamide, enviomycin, para-aminosalicylic acid, cycloserine, amikacin, levofloxacin, moxifloxacin, Gatifloxacin, ofloxacin, terizidone, thionamide, ethionamide, protionamide, clofazimine, linezolid, ampicillin, amoxicillin, clavulanate, thioacetazone, imipenem, cilastatin, bedaquiline, delamanid, rimipenem, cilastatin, meropenem, carbenicillin, chlorampenicol, tetracycline, doxycyline, tigecycline, It may be one or more selected from the group consisting of minocycline, and Collistin.

[0056]

[0057] Another aspect provides a pharmaceutical composition for preventing or treating bacterial or fungal infections, comprising VLX600 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0058] Another aspect provides a method of preventing or treating a bacterial or fungal infection comprising administering VLX600 or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0059] Another aspect provides the use of VLX600 or a pharmaceutically acceptable salt thereof for use in the manufacture of a pharmaceutical composition for the prevention or treatment of a bacterial or fungal infection.

[0060] Another aspect provides the use of VLX600 or a pharmaceutically acceptable salt thereof for the prevention or treatment of a bacterial or fungal infection.

[0061] In one specific example, the pharmaceutical composition may have antibacterial or antifungal activity, or activity for preventing or treating bacterial or fungal infections, against one or more strains selected from the group consisting of Mycobacterium strains, Escherichia strains, Pseudomonas strains, Staphylococcus strains, Acinetobacter strains, and Candida strains.

[0062] Specifically, the pharmaceutical composition for preventing or treating a bacterial infection may have antibacterial activity and activity for preventing or treating a bacterial infection against one or more strains selected from the group consisting of Mycobacterium strains, Escherichia strains, Pseudomonas strains, Staphylococcus strains, and Acinetobacter strains.

[0063] Specifically, the pharmaceutical composition for preventing or treating the fungal infection may have antifungal activity against strains of the genus Candida and activity for preventing or treating the fungal infection.

[0064] In one specific example, the strain of the genus Mycobacterium is Mycobacterium abscessus, Mycobacterium abscessus subsp.abscessus, Mycobacterium abscessus subsp.massiliense, Mycobacterium abscessus subsp.bolletii, Mycobacterium smegmatis, Mycobacterium intracellulare, Mycobacterium avium, Mycobacterium kansasii, Mycobacterium fortuitum, Mycobacterium Mycobacterium gordonae, Mycobacterium osloensis, Mycobacterium phlei, Mycobacterium terrae, Mycobacterium chelonae, Mycobacterium mucogenicum, Mycobacterium peregrinum, Mycobacterium simiae, Mycobacterium wolinskyi, Mycobacterium paragordonae, Mycobacterium ulcerans, Mycobacterium marinum, Mycobacterium bovis, Mycobacterium bovis BCG,It may be at least one selected from the group consisting of Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium canetti, Mycobacterium caprae, Mycobacterium microti, Mycobacterium tuberculosisK strain and Mycobacterium leprae.

[0065] The above Escherichia genus strain may be at least one selected from the group consisting of Escherichia coli, Escherichia albertii, Escherichia blattae, Escherichia fergusonii, Escherichia hermannii, and Escherichia vulneris.

[0066] The above Pseudomonas genus strain may be at least one selected from the group consisting of Pseudomonas aeruginosa, Pseudomonas mendocina, Pseudomonas stutzeri, Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas syringae, Pseudomonas alcaligenes, Pseudomonas luteola, and Pseudomonas oryzihabitans.

[0067] The above Staphylococcus strains include Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Staphylococcus lugdunensis, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus capitis, Staphylococcus warneri, Staphylococcus schleiferi, Staphylococcus intermedius, and Staphylococcus pyogenes. (Staphylococcus pyogens) may be selected from the group consisting of:

[0068] The Acinetobacter genus strain may be at least one selected from the group consisting of Acinetobacter baumannii, Acinetobacter calcoaceticus, Acinetobacter lwoffii, Acinetobacter junii, Acinetobacter haemolyticus, Acinetobacter radioresiststens, Acinetobacter ursingii, Acinetobacter gerneri, and Acinetobacter nosocomialis.

[0069] The Candida genus strain may be at least one selected from the group consisting of Candida albicans, Candida glabrata, Candida tropicalis, Candida pseudotropicalis, Candida parapsilosis, Candida auris, Candida krusei, Candida glabrata, Candida dubliniensis, Candida lusitaniae, Candida kefyr, Candida haemulonii, and Candida rugosa.

[0070] The term "bacterial infection" as used herein refers to a disease caused by bacteria, encompassing a variety of symptoms and diseases caused by bacterial infection. "Bacterial infection" encompasses "bacterial infectious diseases."

[0071] The term "fungal infection" in this specification refers to a disease caused by bacteria, encompassing a variety of symptoms and diseases caused by fungal infections. "Fungal infection" includes "fungal infectious diseases."

[0072] In one specific example, the bacterial infection may be at least one selected from the group consisting of nontuberculous mycobacterial infection, nontuberculous mycobacterial pulmonary disease, tuberculosis, leprosy, enterohemorrhagic Escherichia coli infection, enteropathogenic Escherichia coli infection, multidrug-resistant Pseudomonas aeruginosa infection, multidrug-resistant Acinetobacter baumannii infection, sepsis, pneumonia, enteritis, skin abscess, bloodstream infection, wound infection, cystic fibrosis, mucositis, urinary tract infection, liver abscess, otitis media, keratitis, endophthalmitis, bacteremia, meningitis, cellulitis, and peritonitis.

[0073] In one specific example, the fungal infection may be a systemic infection or an opportunistic infection.

[0074] Specifically, the fungal infection may be at least one selected from the group consisting of candidiasis, oral candidiasis, esophageal candidiasis, vaginal candidiasis, invasive candidiasis, candidemia, candida sepsis, candida bacteremia, candida pneumonia, candida pyelonephritis, candida urinary tract infection, candida endocarditis, candida arthritis, candida osteomyelitis, candida cellulitis, candida peritonitis, candida endophthalmitis, candida keratitis, candida otitis media, and candida meningitis.

[0075] In one specific example, the pharmaceutical composition may additionally comprise an antibiotic.

[0076] Specifically, the pharmaceutical composition for preventing or treating the bacterial infection may additionally contain an antibiotic.

[0077] The above antibiotics include clarithromycin, polymyxin B, erythromycin, isoniazid, rifampicin, ethambutol, SQ-109, pyrazinamide, streptomycin, gentamicin, kanamycin, apramycin, capreomycin, ethionamide, prothionamide, enviomycin, para-aminosalicylic acid, cycloserine, amikacin, levofloxacin, moxifloxacin, Gatifloxacin, ofloxacin, terizidone, thionamide, ethionamide, protionamide, clofazimine, linezolid, ampicillin, amoxicillin, clavulanate, thioacetazone, imipenem, cilastatin, bedaquiline, delamanid, rimipenem, cilastatin, meropenem, carbenicillin, chlorampenicol, tetracycline, doxycyline, tigecycline, It may be one or more selected from the group consisting of minocycline, and Collistin.

[0078] The term "comprising as an active ingredient" in this specification means including an effective amount capable of exhibiting the preventive or therapeutic effect of the pharmaceutical composition on bacterial or fungal infections.

[0079] As used herein, the term “pharmaceutically acceptable” means a substance that can be effectively used for a desired purpose without causing excessive toxicity, irritation, or allergic reaction, within the scope of pharmaceutical judgment.

[0080] As used herein, the term "pharmaceutically acceptable salt" means a salt according to one aspect of the present invention which is pharmaceutically acceptable and has the desired pharmacological activity of the parent compound. Salts of the parent compound can be synthesized from parent compounds containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid form of these compounds with a stoichiometric amount of a suitable base, for example, sodium, calcium, magnesium or potassium, or by reacting the free base form of these compounds with a stoichiometric amount of a suitable acid. These reactions are typically carried out in water or in an organic solvent or in a mixture of the two. Generally, when practical, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile can be used. The pharmaceutically acceptable salts include both addition salts of acids or bases and stereochemically isomeric forms thereof, and may be, for example, addition salts of organic or inorganic acids. The above salt includes any salt that maintains the activity of the parent compound in the subject of administration and does not cause undesirable effects, and is not particularly limited thereto.

[0081] These salts include inorganic and organic salts, for example, acetic acid, nitric acid, aspartic acid, sulfonic acid, sulfuric acid, maleic acid, glutamic acid, formic acid, succinic acid, phosphoric acid, phthalic acid, tannic acid, tartaric acid, hydrobromic acid, propionic acid, benzenesulfonic acid, benzoic acid, stearic acid, lactic acid, bicarboxylic acid, bisulfuric acid, bitartaric acid, oxalic acid, butyric acid, calcium idet, carbonic acid, chlorobenzoic acid, citric acid, idetic acid, toluenesulfonic acid, fumaric acid, gluceptic acid, esilinic acid, pamoic acid, gluconic acid, methylnitric acid, malonic acid, hydrochloric acid, hydroiodoic acid, hydroxynaphtholic acid, isethionic acid, lactobionic acid, mandelic acid, mucic acid, It can be naphthylic acid, muconic acid, p-nitromethanesulfonic acid, hexamic acid, pantothenic acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, salicylic acid, sulfamic acid, sulfanilinic acid, methanesulfonic acid. In addition, the salt form includes salts of alkali and alkaline earth metals such as ammonium salt, lithium salt, sodium salt, potassium salt, magnesium salt, and calcium salt, salts with organic bases such as benzathine, N-methyl-D-glucamine, and hydrabamine salts, and salts with amino acids such as arginine and lysine. In addition, the salt form can be converted into a free form by treating with a suitable base or acid.

[0082] In this specification, the term “prevention” means any action that inhibits or delays the onset of a target disease, “treatment” means any action that improves or beneficially changes a target disease and its metabolic abnormality symptoms by administering a pharmaceutical composition according to the present invention, and “improvement” means any action that reduces a parameter related to a target disease, for example, the severity of a symptom, by administering a composition according to the present invention.

[0083] In one specific example, the pharmaceutical composition may be administered in combination with an antibiotic. The antibiotic is as described above.

[0084] The term “combination administration” as used herein refers to administering the individual components of a treatment regimen simultaneously, sequentially, in reverse order, or individually. It refers to obtaining a combined therapeutic effect by administering two or more drugs simultaneously, sequentially, or in reverse order, or by administering them alternately at regular or indefinite intervals. Combination therapy is not limited thereto, but can be defined as providing a synergistic effect while providing efficacy that is therapeutically superior to the efficacy that can be obtained by administering one or the other of the components of the combination therapy at the usual dose, as measured by, for example, the degree of response, the rate of response, the time until disease progression, or the duration of survival. Combination administration includes simultaneous or sequential administration in any order.

[0085] As used herein, the combination therapy of VLX600 and an antibiotic according to the present invention can provide a "synergistic effect," i.e., the effect achieved when the active substances are used together is greater than the sum of the effects achieved when the active substances are used individually. The synergistic effect can be achieved when the active substances are (1) co-formulated and combined in a unit dosage formulation and administered or delivered simultaneously; (2) delivered sequentially, alternately, or in parallel as separate formulations; or (3) by some other route of administration. When delivered in alternating therapy, the synergistic effect can be achieved, for example, by sequential administration or delivery of the active substances by each injection using a separate syringe. A "synergistic combination" produces an effect that is superior to the sum of the effects of the individual active substances of the combination.

[0086] Combination therapy can provide an "additive" effect, meaning that the effect achieved when the active substances are used together is equal to the sum of the effects achieved when the active substances are used separately.

[0087] The terms "subject" and "patient" are used interchangeably herein. The subject may be an animal. In some embodiments, the subject is a mammal, such as a non-human animal (e.g., a cow, pig, horse, cat, dog, rat, mouse, monkey, or other primate). In some embodiments, the subject is a cynomolgus monkey. In some embodiments, the subject is a human.

[0088] As used herein, the term "therapeutically effective amount" means an amount of a drug, e.g., VLX600, effective to achieve a desired therapeutic or prophylactic result. In some cases, the desired result is treatment of a disease or disorder in a subject. The level of a therapeutically effective amount can be determined based on factors including the type and severity of the patient's disease, the activity and sensitivity of the drug to the drug, the time of administration, the route and excretion rate, the duration of treatment, concomitant medications, and other factors well known in the medical field. The compositions of the present disclosure can be administered as individual therapeutic agents or in combination with other therapeutic agents, can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered in single or multiple doses. That is, the total effective amount of the compositions of the present disclosure can be administered to a patient as a single dose, or can be administered in a fractionated treatment protocol in which multiple doses are administered over a long period of time. Taking all of the above factors into account, it is important to administer an amount that achieves the maximum effect with the minimum amount without causing side effects, and this can be readily determined by those skilled in the art.

[0089] As used herein, terms such as "treating," "treatment," "to treat," "palliating," or "to palliate" refer to therapeutic measures aimed at curing, slowing, alleviating symptoms, and / or arresting the progression of a diagnosed pathological condition or disorder. Therefore, those requiring treatment include those who have already been diagnosed with or are suspected of having a disorder.

[0090] Meanwhile, the pharmaceutical composition of the present specification may additionally include a pharmaceutically acceptable carrier and may be formulated together with the carrier.

[0091] As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier or diluent that does not stimulate a living organism and does not inhibit the biological activity and properties of the administered compound. In a composition formulated as a liquid solution, acceptable pharmaceutical carriers include those that are sterile and biocompatible, such as saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and a mixture of one or more of these components. If necessary, other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate the composition into injectable formulations such as aqueous solutions, suspensions, and emulsions, pills, capsules, granules, or tablets.

[0092] The pharmaceutical composition according to one embodiment of the present disclosure and the composition comprising a pharmaceutically acceptable carrier can be applied to any dosage form containing the pharmaceutical composition as an active ingredient, and can be prepared as an oral or parenteral dosage form, and can be formulated in a unit dosage form for ease of administration and uniformity of dosage. The pharmaceutical dosage form of the present disclosure includes a form suitable for oral, rectal, nasal, topical (including buccal and sublingual), subcutaneous, vaginal, or parenteral (including intramuscular, subcutaneous, and intravenous) administration, or a form suitable for administration by inhalation or insufflation.

[0093] Oral administration dosage forms containing the composition of the present specification as an active ingredient may be formulated, for example, as tablets, troches, lozenges, aqueous or oily suspensions, prepared powders or granules, emulsions, hard or soft capsules, syrups or elixirs.

[0094] The composition of the present disclosure may be formulated as a parenteral dosage form containing the active ingredient, such as a subcutaneous injection, intravenous injection, or intramuscular injection; a suppository injection; or a spray formulation such as an aerosol that can be inhaled through the respiratory tract. To formulate the composition of the present disclosure as an injectable dosage form, the composition of the present disclosure may be mixed with a stabilizer or buffer in water to prepare a solution or suspension, which may then be formulated into a unit dosage form in an ampoule or vial.

[0095] The dosage of the pharmaceutical composition of this specification varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity. The daily dosage is preferably 0.01 μg to 100 mg per kg of body weight per day when administered parenterally, more preferably 1 μg to 50 mg. However, since the dosage may increase or decrease depending on the route of administration, severity of obesity, sex, body weight, age, etc., the above dosage does not limit the scope of this specification in any way.

[0096]

[0097] Another aspect provides a health functional food for preventing or improving bacterial or fungal infections, comprising VLX600.

[0098] Another aspect provides the use of VLX600 for use in the manufacture of a health functional food for the prevention or amelioration of bacterial or fungal infections.

[0099] Another aspect provides for the use of VLX600 for the prevention or amelioration of bacterial or fungal infections.

[0100] In the health functional food of the present invention according to one specific example, “bacteria,” “fungus,” “bacterial infection,” “fungal infection,” “prevention,” and “improvement” are as described in relation to the pharmaceutical composition in this specification.

[0101] In addition, the above health functional food may additionally contain an antibiotic, and the antibiotic is as described above.

[0102] In one specific example, the health functional food may additionally include a food-wise acceptable salt (e.g., a food-wise acceptable salt of VLX600).

[0103] As used herein, the term "food-acceptable salt" refers to a formulation of a compound that does not cause serious irritation to an organism to which the compound is administered and does not impair the biological activity and physical properties of the compound. For example, the food-acceptable salt can be obtained by reacting the compound with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, sulfonic acid such as methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, tartaric acid, formic acid, citric acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, capric acid, isobutanoic acid, malonic acid, succinic acid, phthalic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, salicylic acid, and the like. In addition, it can be obtained by reacting the compound with a base to form salts such as ammonium salts, alkali metal salts such as sodium or potassium salts, alkaline earth metal salts such as calcium or magnesium salts, salts of organic bases such as dicyclohexylamine, N-methyl-D-glucamine, tris(hydroxymethyl)methylamine, and salts of amino acids such as arginine and lysine, but is not limited thereto.

[0104] The health functional food herein may be formulated into any one form selected from the group consisting of powders, tablets, capsules, pills, granules, and liquids, using conventional methods known in the art, but is not limited thereto. Various forms may be manufactured using methods known in the art.

[0105] Additionally, it can be prepared in the form of a composition by mixing with a known substance or active ingredient known to have an activity of preventing, improving or treating bacterial or fungal infections.

[0106] In addition, the health functional food of the present invention may contain conventional food additives, and the suitability as the "food additive" is determined by the specifications and standards for the relevant item according to the general provisions and general test methods of the Food Additives Codex approved by the Ministry of Food and Drug Safety, unless otherwise specified. Items listed in the "Food Additives Codex" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigment, licorice extract, crystalline cellulose, sorghum pigment, and guar gum; and mixed preparations such as sodium L-glutamate preparations, alkaline agents added to noodles, preservative preparations, and tar color preparations.

[0107] In addition to the above, the health functional food of the present invention may include various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the composition of the present invention may include fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These ingredients may be used independently or in combination.

[0108]

[0109] Another aspect provides an antibacterial or antifungal over-the-counter drug product comprising VLX600.

[0110] Another aspect provides the use of VLX600 for the manufacture of antibacterial or antifungal over-the-counter drugs.

[0111] According to one specific example, the antibacterial or antifungal quasi-drug of the present invention has antibacterial or antifungal activity, and in the antibacterial or antifungal quasi-drug of the present invention, “antibacterial,” “fungal,” “antibacterial activity,” and “antifungal activity” are as described in relation to the antibacterial or antifungal composition herein.

[0112] In addition, the above antibacterial or antifungal over-the-counter drug may additionally contain an antibiotic, and the antibiotic is as described above.

[0113] In this specification, the term "quasi-drug" refers to products that are used for the purpose of diagnosing, treating, improving, alleviating, treating, or preventing diseases in humans or animals, and have a milder effect than pharmaceuticals. Specifically, it refers to products other than those used for pharmaceutical purposes, such as fibers, rubber products, or similar products; products that have a weak effect on the human body or do not directly affect the human body, and are not devices or machines or similar products; and products used for sterilization, insecticide, and similar purposes to prevent infection. In other words, it may refer to products that are not devices, machines, or devices among products used for the purpose of diagnosing, treating, alleviating, treating, or preventing diseases in humans or animals, and products that are not devices, machines, or devices among products used for the purpose of exerting a pharmacological effect on the structure and function of humans or animals. Quasi-drugs also include external skin preparations and personal hygiene products.

[0114] The antibacterial or antifungal over-the-counter drug containing VLX600 according to the present invention can be used in combination with other over-the-counter drug ingredients and can be used appropriately according to conventional methods. The mixing amount of the active ingredients can be appropriately determined depending on the intended use.

[0115] The over-the-counter drug of the present invention may be manufactured and used in the form of a cream, lotion, aerosol, shampoo, gel, or pack, but is not limited thereto.

[0116] In the case of the above creams, ointments, shampoos, gels or packs, bases such as white petrolatum, yellow petrolatum, lanolin, bleached beeswax, cetanol, stearyl alcohol, stearic acid, hydrogenated oils, gelling hydrocarbons, polyethylene glycol, liquid paraffin, squalane, etc.; solvents and solubilizing agents such as oleic acid, isopropyl myristate, glycerin triisooctanoate, crotamiton, diethyl sebacate, diisopropyl adipate, hexyl laurate, fatty acids, fatty acid esters, aliphatic alcohols, vegetable oils, etc.; antioxidants such as tocopherol derivatives, L-ascorbic acid, dibutylhydroxytoluene, butylhydroxyanisole, etc.; preservatives such as parahydroxybenzoate esters, etc.; moisturizers such as glycerin, propylene glycol, sodium hyaluronate, etc. Surfactants such as polyoxyethylene derivatives, glycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, and lecithin; thickeners such as carboxyvinyl polymers, xanthan gum, carboxymethylcellulose, carboxymethylcellulose sodium salts, hydroxypropylcellulose, and hydroxypropylmethylcellulose.

[0117]

[0118] Another aspect provides an antibacterial or antifungal cosmetic composition comprising VLX600.

[0119] Another aspect provides the use of VLX600 for the preparation of an antibacterial or antifungal cosmetic composition.

[0120] According to one specific example, the antibacterial or antifungal cosmetic composition of the present invention has antibacterial or antifungal activity, and in the antibacterial or antifungal cosmetic composition of the present invention, “antibacterial,” “fungal,” “antibacterial activity,” and “fungal activity” are as described herein with respect to the antibacterial or antifungal composition.

[0121] In addition, the above-mentioned antibacterial or antifungal cosmetic composition may additionally contain an antibiotic, which is as described above.

[0122] In one specific embodiment, the cosmetic composition may further include additional ingredients commonly used in cosmetics. For example, these may include conventional adjuvants and carriers, such as stabilizers, solubilizers, vitamins, pigments, and fragrances. Those skilled in the art can select any additional ingredients and / or their amounts so that the beneficial properties of the composition according to the present disclosure are not adversely affected or substantially affected by the anticipated addition.

[0123] In one specific example, the cosmetic composition may be applied alone or in combination, or may be applied in combination with other cosmetic compositions other than those of the present invention. Furthermore, the cosmetic composition according to the present invention may be used according to conventional methods, and the frequency of use may vary depending on the user's skin (or scalp) condition or preference.

[0124] The cosmetic composition of the present invention can be manufactured in any formulation commonly manufactured in the art, and can be manufactured as, for example, emulsion, cream, toner, pack, foundation, lotion, beauty liquid, hair cosmetic, etc. Specifically, the cosmetic composition of the present invention includes formulations of skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, essence, nutrition essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, and body cleanser. In addition, it includes formulations of hair tonic, hair cream, hair lotion, hair shampoo, hair rinse, hair conditioner, hair spray, hair aerosol, pomade, powder gel, hair pack, hair treatment, eyebrow hair tonic, eyelash hair tonic, or eyelash nutrient.

[0125] When the formulation of the present invention is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component.

[0126] When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and particularly in the case of a spray, a propellant such as chlorofluorohydrocarbon, propane / butane or dimethyl ether may be additionally included.

[0127] In the case where the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

[0128] When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth may be used as carrier components.

[0129] When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolenic derivative, or ethoxylated glycerol fatty acid ester may be used as a carrier component.

[0130]

[0131] Another aspect provides an antibacterial or antifungal feed composition comprising VLX600.

[0132] Another aspect provides the use of VLX600 for the preparation of an antibacterial or antifungal feed composition.

[0133] According to one specific example, the antibacterial or antifungal feed composition of the present invention has antibacterial or antifungal activity, and in the antibacterial or antifungal feed composition of the present invention, “antibacterial,” “fungal,” “antibacterial activity,” and “antifungal activity” are as described herein with respect to the antibacterial or antifungal composition.

[0134] Additionally, the antibacterial or antifungal feed composition may additionally contain an antibiotic, which is as described above.

[0135] In this specification, “feed composition” may mean any natural or artificial diet, meal, etc. or a component of said meal, especially for or suitable for eating, ingesting and digesting by an animal.

[0136] The above feed composition may include, but is not particularly limited to, nutrients such as energy, protein, lipids, vitamins, and minerals required by the individual consuming the feed. The individual refers to the subject of breeding, and includes, without limitation, any living organism capable of consuming the feed of the present invention, including companion animals and livestock.

[0137] The type of the above feed is not particularly limited, and feed commonly used in the relevant technical field can be used. Non-limiting examples of the above feed include plant-based feed such as grains, roots, food processing by-products, algae, fiber, pharmaceutical by-products, oils, starches, meal, or grain by-products; and animal-based feed such as proteins, inorganic substances, oils, minerals, oils, single-cell proteins, zooplankton, or food. These may be used alone or in combination of two or more types.

[0138] The feed composition according to the present invention can be manufactured by adding VLX600 of the present invention in an appropriate effective concentration range according to various feed manufacturing methods known in the art.

[0139] The feed composition according to the present invention can be applied to any individual for which the antibacterial effect according to the present invention is desired, without limitation. For example, it can be applied to any individual, including non-human animals such as monkeys, dogs, cats, rabbits, guinea pigs, rats, mice, cows, sheep, pigs, goats, birds, and fish.

[0140] An antibacterial or antifungal composition comprising VLX600 according to one aspect of the present invention has antibacterial or antifungal activity against a wide range of strains, including Mycobacterium strains, Escherichia strains, Pseudomonas strains, Staphylococcus strains, Acinetobacter strains, and Candida strains, and a pharmaceutical composition comprising VLX600 has excellent antibacterial, antifungal, and therapeutic effects against pathogens, and can be usefully utilized for the prevention or treatment of bacterial or fungal infections. In addition, VLX600 has a synergistic effect with existing antibiotics, and is effective in treating infections caused by antibiotic-resistant strains.

[0141] Figure 1 is a schematic diagram of the production of luminescent recombinant strain Mab-luxG13 and the evaluation of antibacterial efficacy of drugs using it.

[0142] Figure 2 shows the process of discovering VLX600 through screening and the chemical structure of VLX600.

[0143] Figure 3 shows the results of the safety evaluation of VLX600 in vitro using the NRU (Neutral red uptake) assay.

[0144] Figure 4 is a schematic diagram of the MIC measurement of VLX600 using the microdilution technique.

[0145] Figure 5 is a graph evaluating the direct antibacterial efficacy of VLX600 against the M. abscessussubsp.abscessus strain through luminescence measurement.

[0146] Figure 6 is a graph evaluating the direct antibacterial efficacy of VLX600 against the M. abscessussubsp.massiliense strain through luminescence measurement.

[0147] Figure 7 is a graph evaluating the direct antibacterial efficacy of VLX600 against three standard strains of M. abscessus through MIC measurement.

[0148] Figure 8 is a graph evaluating the antibacterial efficacy of VLX600 against M. abscesuss in macrophages through luminescence measurement.

[0149] Figure 9 is a graph evaluating the antibacterial efficacy of VLX600 against M. abscesuss in macrophages through luminescence measurement.

[0150] Figure 10 is a graph evaluating the direct antibacterial efficacy of VLX600 against Mycobacteria species other than M. abscessus through MIC measurement.

[0151] Figure 11 is a graph evaluating the direct antibacterial efficacy of VLX600 against species other than Mycobacteria through MIC measurement.

[0152] Figure 12 is a graph evaluating the antibacterial efficacy of VLX600 against A. baumannii in lung cells and macrophages through CFU analysis.

[0153] Figure 13 is a schematic diagram of the in vivo therapeutic efficacy evaluation of VLX600 using the M. abscessus respiratory infection model.

[0154] Figure 14 is a schematic diagram of the in vivo therapeutic efficacy evaluation of VLX600 using an A. baumannii sepsis model.

[0155] Figure 15 is a schedule for evaluating the efficacy of VLX600 in the M. abscessus respiratory infection model.

[0156] Figure 16 is a graph evaluating changes in mouse body weight and CFU in lung tissue following M. abscessus infection.

[0157] Figure 17 is a photograph showing inflammation in lung tissue through H&E staining.

[0158] Figure 18 is a graph evaluating the improvement in survival rate by VLX600 administration in the A. baumannii sepsis model.

[0159] Figure 19 is a schematic diagram for evaluating the synergistic effect of VLX600 and antibiotics using SynergyFinder.

[0160] Figures 20 and 21 show the results of evaluating the synergistic effect of VLX600 and Clarithromycin.

[0161] Figures 22 and 23 show the results of evaluating the synergistic effect of VLX600 and Polymyxin B.

[0162] Hereinafter, preferred embodiments are presented to aid understanding of the present invention. However, the following embodiments are provided solely to facilitate a better understanding of the present invention and are not intended to limit the scope of the present invention. The embodiments are susceptible to various modifications, and thus the embodiments are not limited to the embodiments disclosed below and may be implemented in various forms.

[0163] Terms or words used in the specification and claims of the present invention are not to be construed as limited to their usual or dictionary meanings, and should be interpreted as meanings and concepts that conform to the technical idea of ​​the present invention based on the principle that the inventor can appropriately define the concept of the term to explain his or her own invention in the best way.

[0164] Throughout the specification of the present invention, when a part is said to "include" a certain component, this does not mean that other components are excluded, but rather that other components may be included, unless specifically stated otherwise.

[0165] Throughout the specification of the present invention, “A and / or B” means A or B, or A and B.

[0166]

[0167] Example 1. Discovery of new antibiotics through drug repurposing.

[0168] To discover novel antibiotics through drug repurposing, a recombinant strain (Mab-luxG13) expressing luciferase was constructed from M. abscessus isolated from a patient with lung infection, and a screening system was established. The produced Mab-luxG13 exhibits luminescence that increases in proportion to the number of viable bacteria and decreases when viability is reduced by heat treatment or antibiotics. Therefore, the antibacterial activity of candidate substances can be efficiently evaluated by measuring luminescence.

[0169]

[0170] Figure 1 is a schematic diagram of the production of luminescent recombinant strain Mab-luxG13 and the evaluation of antibacterial efficacy of drugs using it.

[0171]

[0172] A compound library consisting of 3,200 clinical compounds was directly treated with Mab-luxG13 and treated with a macrophage infection model to confirm antibacterial efficacy by luminescence, and “VLX600”, which showed high antibacterial efficacy in both models, was selected.

[0173]

[0174] Figure 2 shows the process of discovering VLX600 through screening and the chemical structure of VLX600.

[0175]

[0176] Example 2. Evaluation of antibacterial and antifungal activity of VLX600

[0177] The antibacterial activity of VLX600 in direct treatment and macrophage infection models was confirmed according to the concentration of VLX600, and this was confirmed not only through luminescence but also through the CFU assay, a commonly used technique. In addition, the cytotoxicity in macrophages according to VLX600 treatment was confirmed through the NRU (Neutral red uptake) assay, confirming the safety of VLX600 in vitro.

[0178]

[0179] Figure 3 shows the results of the safety evaluation of VLX600 in vitro using the NRU (Neutral red uptake) assay.

[0180]

[0181] In addition, the antibacterial activity against other species of bacteria, such as M. intracellulare, M. bovisBCG, M. smegmatis, and E. coli, P. aeruginosa, A. baumannii, S. aureus, and C. albicans, was also evaluated by measuring the MIC using the microdilution technique.

[0182]

[0183] Figure 4 is a schematic diagram of the MIC measurement of VLX600 using the microdilution technique.

[0184]

[0185] The antibacterial activity evaluation of VLX600 is described in detail through Experimental Examples 1 to 6 below.

[0186]

[0187] Experimental Example 1. Direct confirmation of the antibacterial efficacy of VLX600 against M. abscessus through luminescence measurement.

[0188] The efficacy was confirmed by measuring luminescence as follows. Mab-luxG13 or Mmas-luxG13 cultured at 37°C to the stationary phase was dissolved in 1% Tween-80 solution to remove clumps, and then incubated in complete 7H9 culture medium (2.5% glycerol, 0.5% Tween-80, 10% ADC in 7H9) at OD 600nm Diluted to =0.2.

[0189] Mab-luxG13 is a strain recombinantly modified to express luciferase in M. abscessussubsp.abscessus, and Mmas-luxG13 is a strain recombinantly modified to express luciferase in M. abscessussubsp.massiliense.

[0190] LumiNunc diluted bacterial culture TM 100 μl each was added to a 96-well plate (Thermo 437796), and 100 μl each was treated with VLX600 and clarithromycin diluted in complete 7H9 to twice the target concentrations of 10 μM, 5 μM, and 2.5 μM. After treatment, culture was performed in a 37°C incubator, and the luminescence was measured using a Tecan F200 microplate reader at 0, 24, 48, and 72 hours after treatment.

[0191]

[0192] Figure 5 is a graph evaluating the direct antibacterial efficacy of VLX600 against the M. abscessussubsp.abscessus strain through luminescence measurement.

[0193] Figure 6 is a graph evaluating the direct antibacterial efficacy of VLX600 against the M. abscessussubsp.massiliense strain through luminescence measurement.

[0194]

[0195] As shown in FIGS. 5 and 6, unlike clarithromycin, which did not significantly reduce the viability of Mab-luxG13 and Mmas-luxG13 compared to the negative control group DMSO due to drug resistance, VLX600 significantly inhibited the viability of Mab-luxG13 and Mmas-luxG13 depending on the concentration and time.

[0196]

[0197] Experimental Example 2. Direct confirmation of the antibacterial efficacy of VLX600 against M. abscessus through MIC measurement.

[0198] Efficacy was confirmed through MIC measurement according to the guidelines presented by the Clinical & Laboratory Standards Institute (CLSI). Three standard strains of M. abscessus (subsp. Abscessus: ATCC 19977, subsp. Massiliense: KCTC 19086, subsp. Bolteii: KCTC19281) cultured at 37℃ to the stationary phase were dissolved in 0.1% Tween-80 solution to remove clumps, and then diluted in PBS to a McFarland index of 0.5. This was diluted 1:100 in Mueller-Hinton solution, 100 μl each was added to a 96-well plate, and VLX600 was serially diluted in Mueller-Hinton solution to twice the target concentration, and 100 μl each was treated. After treatment, the culture was incubated for one week in a 37℃ incubator, and OD600nm was measured using a Tecan F200 microplate reader. MIC was defined as the concentration at which growth was inhibited by 90% or more compared to the untreated control group.

[0199]

[0200] Figure 7 is a graph evaluating the direct antibacterial efficacy of VLX600 against three standard strains of M. abscessus through MIC measurement.

[0201]

[0202] As shown in Fig. 7, VLX600 effectively inhibited the growth of M. abscessus depending on the concentration, as confirmed by OD600nm measurement, and the efficacy was maintained stably without disappearing even after one week.

[0203] In addition, it showed excellent inhibitory effects on all three subspecies of M. abscessussubsp.abscessus, M. abscessussubsp.massiliense, and M. abscessussubsp.bolletii, and reached MIC at 8 μg / ml for M. abscessussubsp.abscessus, 4 μg / ml for M. abscessussubsp.massiliense, and 2 μg / ml for M. abscessussubsp.bolletii, indicating that susceptibility to VLX600 is high in the order of bolletii, massiliense, and abscessus.

[0204]

[0205] Experimental Example 3. Confirmation of the antibacterial efficacy of VLX600 against M. abscesus in macrophages using luminescence measurements.

[0206] The antibacterial efficacy of VLX600 against M. abscessus in macrophages was evaluated using the Mab_luxG13-macrophage infection model using the macrophage cell line J774A.1. 1x10 5 J774A.1 of LumiNunc TM96-well plates (Thermo 437796) were seeded, cultured overnight, and starved in Opti-MEM for 1 hour. VLX600 was diluted at various concentrations in RPMI-1640 containing 2% FBS and pretreated J774A.1 for 16 hours. After removing the culture medium containing the compound, Mab_luxG13 was infected into J774A.1 at an MOI of 10 for 2 hours. The infected Mab_luxG13 was removed, washed with PBS, and cultured in RPMI-1640 containing 50 μg / ml amikacin for approximately 1 hour to inactivate Mab_luxG13 that had not entered the cells. After removing the culture medium containing amikacin, the candidate compounds were diluted at various concentrations in RPMI-1640 containing 2% FBS and cultured, and luminescence was measured after 24 hours using a Tecan F200 Microplater Reader machine.

[0207]

[0208] Figure 8 is a graph evaluating the antibacterial efficacy of VLX600 against M. abscesuss in macrophages through luminescence measurement.

[0209]

[0210] As shown in Fig. 8, VLX600 was confirmed to effectively inhibit the growth of M. abscessus within macrophage cells. Furthermore, this effect was confirmed not only in the Rough strain, which is one of two colony phenotypes of M. abscessus, but also in the Smooth strain, and was confirmed to be effective in the massiliense subspecies of M. abscessus.

[0211]

[0212] Experimental Example 4. Confirmation of the antibacterial efficacy of VLX600 against M. abscesus in macrophages through CFU analysis.

[0213] CFU analysis was performed with 1x10 in each well. 5Macrophage cell line J774A.1 was seeded in a 96-well cell culture plate and cultured for 12 to 16 hours, followed by starvation in Opti-MEM for 1 to 2 hours. VLX600 was diluted at various concentrations in RPMI-1640 containing 2% FBS and pretreated to J774A.1 for 12 to 16 hours. After removing the culture medium containing the compound, M. abscessus was infected into J774A.1 at an MOI of 10 for 2 hours. The infected M. abscessus was removed, washed with PBS, and cultured in RPMI-1640 containing 50 μg / ml amikacin for approximately 1 hour to inactivate M. abscessus that had not entered the cells. After that, the culture medium containing amikacin was removed, and the candidate compounds were diluted at various concentrations and cultured in RPMI-1640 containing 2% FBS. After 24 hours, they were washed with PBS and J774A.1 was lysed by adding PBS containing 1% Triton X-100. The lysed solution was serially diluted with PBS, dropped 20 μl at a time onto 7H10 agar medium, and cultured at 37°C until colonies were formed, and the formed colonies were counted.

[0214]

[0215] Figure 9 is a graph evaluating the antibacterial efficacy of VLX600 against M. abscesuss in macrophages through luminescence measurement.

[0216]

[0217] As shown in Fig. 9, VLX600 effectively inhibited the growth of M. abscessus within macrophages, as confirmed not only by luminescence measurements but also by actual CFU counts. Furthermore, this effect was confirmed not only in the Rough strain, but also in the Smooth strain, two colony phenotypes of M. abscessus, and was confirmed to work effectively in the massiliense subspecies of M. abscessus.

[0218]

[0219] Experimental Example 5. Confirmation of the antibacterial efficacy of VLX600 against other Mycobacteria species.

[0220] The antibacterial efficacy of VLX600 against different Mycobacteria strains was evaluated using MIC measurements. Efficacy was confirmed using MIC measurements in accordance with the guidelines presented by the Clinical & Laboratory Standards Institute (CLSI).

[0221] M. smegmatis (ATCC 700084), M. bovis BCG (strain Tokyo), and M. intracellulare (ATCC 13950), cultured to the stationary phase at 37℃, were diluted in PBS to a McFarland index of 0.5. These were diluted 1:100 in Mueller-Hinton solution and 100 μl each was added to a 96-well plate. VLX600 was serially diluted in Mueller-Hinton solution to twice the target concentration and treated with 100 μl each. After treatment, M. smegmatis was cultured for 3 days in an incubator at 37℃, and M. bovis BCG and M. intracellulare were cultured for 7 days, and the OD600 nm was measured using a Tecan F200 microplate reader. The MIC was defined as the concentration at which growth was inhibited by 90% or more compared to the untreated control.

[0222]

[0223] Figure 10 is a graph evaluating the direct antibacterial efficacy of VLX600 against Mycobacteria species other than M. abscessus through MIC measurement.

[0224]

[0225] As shown in Fig. 10, VLX600 exhibited growth inhibition effects depending on the concentration when directly treated with other mycobacteria species, such as M. smegmatis, M. bovis BCG, and M. intracellulare. The MIC was reached at 4 μg / ml for M. smegmatis and M. intracellulare, and at 8 μg / ml for M. bovis BCG. These results confirmed that VLX600 possesses high direct antibacterial efficacy against mycobacteria other than M. abscessus.

[0226]

[0227] Experimental Example 6. Confirmation of the antibacterial efficacy of VLX600 against species other than Mycobacteria.

[0228] The antibacterial efficacy of VLX600 against bacterial and fungal species other than Mycobacteria was evaluated using MIC measurements. Efficacy confirmation through MIC measurements was conducted in accordance with the guidelines presented by the Clinical & Laboratory Standards Institute (CLSI), as follows.

[0229] Escherichia coli (NCCP 14762), Pseudomonas aeruginosa (NCCP 14781), Staphylococcus aureus (NCCP 14780), Acinetobacter baumannii (ATCC 17978 and 2 clinical strains; A0062, A0292), and Candida albicans (NCCP32557) cultured to the stationary phase at 37℃ were diluted in PBS to a McFarland standard of 0.5. These were diluted 1:100 in Mueller-Hinton solution, placed 100 μl each in a 96-well plate, and VLX600 was serially diluted in Mueller-Hinton solution to twice the target concentration, and treated with 100 μl each. After treatment, culture was performed in a 37℃ incubator for 20-24 hours, and OD600nm was measured using a Tecan F200 microplate reader. MIC was defined as the concentration at which growth was inhibited by 90% or more compared to the untreated control group.

[0230]

[0231] Figure 11 is a graph evaluating the direct antibacterial efficacy of VLX600 against species other than Mycobacteria through MIC measurement.

[0232]

[0233] As shown in Fig. 11, VLX600 showed a growth inhibition effect depending on the concentration when directly treated with bacterial strains other than mycobacteria, such as E. coli, P. aeruginosa, S. aureus, and A. baumannii. In addition, VLX600 showed a growth inhibition effect depending on the concentration when directly treated with fungal strains, such as Candida albicans. The MIC was reached at 16 μg / ml for E. coli, 4 μg / ml for P. aeruginosa, 16 μg / ml for S. aureus, 8 μg / ml for A. baumannii, and 2 μg / ml for C. albicans. From these results, it can be determined that VLX600 is a broad-spectrum antibiotic that exhibits antibacterial and antifungal efficacy against various bacterial and fungal species.

[0234]

[0235] Experimental Example 7. Confirmation of the antibacterial efficacy of VLX600 against A. baumannii in lung cells and macrophages through CFU analysis.

[0236] The antibacterial efficacy of VLX600 against intracellular A. baumannii was evaluated using the lung cell line A549 and the macrophage cell line J774A.1 as an A. baumannii-cell infection model. 1x10 5Cells were seeded in 96-well plates, cultured overnight, and starved in Opti-MEM for 1 hour. VLX600 was diluted at various concentrations in RPMI-1640 containing 2% FBS and pretreated J774A.1 for 16 hours. After removing the culture medium containing the compound, A. baumannii was infected with J774A.1 at an MOI of 10 for 4 hours. The infected A. baumannii was removed, washed with PBS, and cultured in RPMI-1640 containing 50 ug / ml amikacin for about 1 hour to inactivate A. baumannii that had not entered the cells. The culture medium containing amikacin was removed, and VLX600 was diluted at various concentrations in RPMI-1640 containing 2% FBS and cultured. After 24 hours, the cells were washed with PBS and lysed with PBS containing 1% Triton X-100. The lysed solution was serially diluted with PBS, dropped 20 μl at a time onto LB agar medium, and cultured at 37°C until colonies were formed, after which the resulting colonies were counted.

[0237]

[0238] Figure 12 is a graph evaluating the antibacterial efficacy of VLX600 against A. baumannii in lung cells and macrophages through CFU analysis.

[0239]

[0240] As shown in Fig. 12, it was confirmed that VLX600 effectively inhibited the growth of A. baumannii present in lung cells and macrophages.

[0241]

[0242] Example 3. Evaluation of the in vivo therapeutic efficacy of VLX600

[0243] To evaluate the in vivo efficacy of VLX600, the following model was constructed and experiments were conducted.

[0244] Specifically, after establishing a M. abscessus mouse respiratory infection model, VLX600 was administered intraperitoneally daily to evaluate the reduction in bacterial counts and the degree of inflammation relief in lung tissue compared to the control group. In addition, after establishing an A. baumannii sepsis model, the improvement in survival rate following VLX600 administration was evaluated.

[0245]

[0246] Figure 13 is a schematic diagram of the in vivo therapeutic efficacy evaluation of VLX600 using the M. abscessus respiratory infection model.

[0247] Figure 14 is a schematic diagram of the in vivo therapeutic efficacy evaluation of VLX600 using an A. baumannii sepsis model.

[0248]

[0249] The in vivo therapeutic efficacy evaluation of VLX600 is described in detail through Experimental Example 8 below.

[0250]

[0251] Experimental Example 8. Confirmation of the antibacterial and therapeutic efficacy of VLX600 in vivo.

[0252] 8.1 In vivo confirmation of the antibacterial and therapeutic efficacy of VLX600 against M. abscessus.

[0253] Eight-week-old female BALB / c mice were used. To increase the infection rate of M. abscessus, 150 mg / kg of cyclophosphamide (Sigma, #PHR1404) was administered intraperitoneally on day 1 and 4 before infection to induce neutropenia in the mice. M. abscessus (Clinical Mab_R), derived from a patient with refractory lung disease, was cultured at 37°C until the stationary phase and administered to mice at a dose of 1 x 10 6Respiratory infections were induced by intranasal administration of CFU (Colony Forming Unit). The PBS administration group served as a negative control group, and the amikacin administration group (50 mg / kg) served as a positive control group. To confirm the efficacy of the VLX600 administration group, 4.5 mg / kg of VLX600 was administered intraperitoneally daily from one day after infection until the 10th day of infection. On the 10th day of infection, the mice were sacrificed, and the lungs were isolated. Some of them were fixed in 10% formalin and stained with H&E for histological examination, and the rest were homogenized in PBS and subjected to CFU analysis on 7H10 solid medium.

[0254]

[0255] Figure 15 is a schedule for evaluating the efficacy of VLX600 in the M. abscessus respiratory infection model.

[0256] Figure 16 is a graph evaluating changes in mouse body weight and CFU in lung tissue following M. abscessus infection.

[0257] Figure 17 is a photograph showing inflammation in lung tissue through H&E staining. In Figure 17, the upper left is the non-infected control group, the upper right is the PBS administration group, the lower left is the Amikacin administration group, and the lower right is the VLX600 administration group.

[0258]

[0259] As shown in Fig. 16, the VLX600-administered group showed weight loss until the third day of administration, and then gradually increased again from the fourth day, resulting in a 3.25% weight loss compared to the PBS-administered control group on the 10th day of infection, which was lower than that of Amikacin (6.84%). This weight loss can be considered to be due to VLX600 administration in addition to the decrease due to M. abscessus infection. When CFU was checked, the CFU in the lung tissue of the VLX600-administered group was reduced by approximately 60% compared to the PBS-administered control group, which was statistically significant. These results indicate that the number of M. abscessus bacteria in the lung tissue was reduced by VLX600.

[0260] As shown in Figure 17, when inflammation in lung tissue was confirmed through histological examination, it was confirmed that inflammation was significantly reduced in the VLX600 administration group compared to the PBS administration control group.

[0261] The above results indicate that VLX600 has excellent antibacterial or antifungal efficacy and therapeutic efficacy against pathogens and can be used as a pharmaceutical composition for preventing or treating bacterial or fungal infections.

[0262]

[0263] 8.2A. In vivo confirmation of the antibacterial and therapeutic efficacy of VLX600 against B. baumannii

[0264] Eight-week-old female BALB / c mice were used, and carbapenem-resistant A. baumannii (A0292) derived from a lung disease patient was cultured at 37°C until the stationary phase and injected into the mice at a dose of 1x10 8 Sepsis was induced by intraperitoneal administration of CFU. VLX600 was administered intraperitoneally at a dose of 20 mg / kg 2 hours before infection, and the PBS group was used as a negative control group, and the survival rate was compared over time.

[0265]

[0266] Figure 18 is a graph evaluating the improvement in survival rate by VLX600 administration in the A. baumannii sepsis model.

[0267]

[0268] As shown in Figure 18, the PBS-administered control group showed a significant decrease in survival rate over time, whereas the VLX600-administered group showed no decrease in survival rate over time.

[0269] The above results indicate that VLX600 has excellent antibacterial or antifungal efficacy and therapeutic efficacy against pathogens and can be used as a pharmaceutical composition for preventing or treating bacterial or fungal infections.

[0270]

[0271] Example 4. Evaluation of the synergistic effect of VLX600 and antibiotics

[0272] Because treatment of highly antibiotic-resistant bacterial infections often involves administering multiple antibiotics in combination, synergy between antibiotics can be a valuable advantage in new drug development. Accordingly, we evaluated the synergistic effects of VLX600 with currently clinically used antibiotics using a web-based analysis program called SynergyFinder.

[0273]

[0274] Figure 19 is a schematic diagram for evaluating the synergistic effect of VLX600 and antibiotics using SynergyFinder.

[0275]

[0276] The evaluation of the synergistic effect of VLX600 and antibiotics is described in detail through Experimental Example 9 below.

[0277]

[0278] Experimental Example 9. Confirmation of the synergistic effect of VLX600 and antibiotics.

[0279] A checkerboard assay was performed to determine whether there was a synergistic effect when Clarithromycin, an antibiotic used to treat M. abscessus infection, and Polymyxin B, an antibiotic used to treat A. baumannii infection, were used together with VLX600.

[0280] Specifically, 1x10 in each well 5 J774A.1 of LumiNunc TM The cells were seeded in 96-well plates (Thermo 437796) and cultured for 16 hours, and Mab_luxG13 was infected with J774A.1 at an MOI of 10 for 2 hours. The culture medium was removed, washed with PBS, and cultured in RPMI-1640 containing 50 μg / ml amikacin for about 1 hour to inactivate bacteria that had not entered the cells. The culture medium containing amikacin was removed, and RPMI-1640 containing 2% FBS and VLX600 and clarithromycin were diluted at different concentrations. Similarly, 100 μl of antibiotics were treated from top to bottom in order of decreasing concentration, and 100 μl of VLX600 were treated from left to right in order of decreasing concentration, and a checkerboard assay was performed. After treatment, luminescence was measured over time using a Tecan F200 Microplate Reader.

[0281] For A. baumannii, the process was performed by directly treating the bacterial culture with Polymyxin B and VLX600. The bacteria were diluted to McFarland standard 0.5 and then diluted 1:100 in Muller-Hinton broth to prepare an inoculum, which was loaded 100 μL into each well. VLX600 and Polymyxin B were also diluted to the target concentrations in Muller-Hinton broth, and then 100 μL of Polymyxin B were treated from the top to the bottom of the plate, starting with the lowest concentration, and 100 μL of VLX600 were treated from the left to the right, starting with the lowest concentration, to perform a checkerboard assay. After treatment, the plates were cultured at 37°C for 16-20 hours, and the OD600 was measured using a Tecan F200 Microplate Reader.

[0282] Analysis of the synergy effect was performed using SynergyFinder Plus (https: / synergyfinder.org), a web-based analysis tool, and a synergy effect was considered to exist if the synergy score was 5 or higher through four calculation methods: ZIP, BLiss, Loewe, and HSA.

[0283]

[0284] Figures 20 and 21 show the results of evaluating the synergistic effect of VLX600 and Clarithromycin.

[0285] Figures 22 and 23 show the results of evaluating the synergistic effect of VLX600 and Polymyxin B.

[0286]

[0287] As shown in Figures 20 and 21, a high synergistic effect was confirmed when Clarithromycin was treated together with VLX600 in Mab-luxG13 infected into macrophages. High scores of 15 points or more were confirmed in all four calculation methods: ZIP, BLiss, Loewe, and HSA.

[0288] As shown in Figures 22 and 23, a high synergistic effect was also confirmed when Polymyxin B and VLX600 were treated together on A. baumannii. High scores that far exceeded the criterion score of 5 points for the synergistic effect were confirmed in all four calculation methods (ZIP, BLiss, Loewe, and HSA).

[0289] This synergistic effect between VLX600 and antibiotics means that it is effective in treating infections caused by antibiotic-resistant strains, and it can be judged that it will be very advantageous in that multiple antibiotics are used in combination in clinical practice to treat infections caused by antibiotic-resistant strains.

[0290]

[0291] From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without altering its technical concept or essential characteristics. In this regard, it should be understood that the experimental examples and embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as encompassing all changes or modifications derived from the meaning and scope of the following claims and their equivalent concepts, rather than the detailed description above.

Claims

1. An antibacterial or antifungal composition comprising VLX600.

2. An antibacterial or antifungal composition according to claim 1, wherein the composition has antibacterial activity against at least one strain selected from the group consisting of Mycobacterium strains, Escherichia strains, Pseudomonas strains, Staphylococcus strains, Acinetobacter strains, and Candida strains.

3. In the second paragraph, the strain of the genus Mycobacterium is Mycobacterium abscessus, Mycobacterium abscessus subsp.abscessus, Mycobacterium abscessus subsp.massiliense, Mycobacterium abscessus subsp.bolletii, Mycobacterium smegmatis, Mycobacterium intracellulare, Mycobacterium avium, Mycobacterium kansasii, Mycobacterium fortuitum, Mycobacterium Mycobacterium gordonae, Mycobacterium osloensis, Mycobacterium phlei, Mycobacterium terrae, Mycobacterium chelonae, Mycobacterium mucogenicum, Mycobacterium peregrinum, Mycobacterium simiae, Mycobacterium wolinskyi, Mycobacterium paragordonae, Mycobacterium ulcerans, Mycobacterium marinum, Mycobacterium bovis, Mycobacterium bovis BCG,An antibacterial or antifungal composition comprising at least one selected from the group consisting of Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium canetti, Mycobacterium caprae, Mycobacterium microti, Mycobacterium tuberculosis K strain, and Mycobacterium leprae.

4. An antibacterial or antifungal composition according to claim 1, wherein the composition further comprises an antibiotic.

5. In paragraph 4, the antibiotic is clarithromycin, polymyxin B, erythromycin, isoniazid, rifampicin, ethambutol, SQ-109, pyrazinamide, streptomycin, gentamicin, kanamycin, apramycin, capreomycin, ethionamide, prothionamide, enviomycin, para-aminosalicylic acid, cycloserine, amikacin, levofloxacin, Moxifloxacin, Gatifloxacin, ofloxacin, terizidone, thionamide, ethionamide, protionamide, clofazimine, linezolid, ampicillin, amoxicillin, clavulanate, thioacetazone, imipenem, cilastatin, bedaquiline, delamanid, rimipenem, cilastatin, meropenem, carbenicillin, chlorampenicol, tetracycline, doxycycline (doxycyline), tigecycline, minocycline,An antibacterial composition comprising at least one selected from the group consisting of and Collistin.

6. A pharmaceutical composition for preventing or treating bacterial or fungal infections, comprising VLX600 or a pharmaceutically acceptable salt thereof as an active ingredient.

7. A pharmaceutical composition according to claim 6, wherein the bacteria is at least one selected from the group consisting of Mycobacterium strains, Escherichia strains, Pseudomonas strains, Staphylococcus strains, Acinetobacter strains, and Candida strains.

8. In the 7th paragraph, the strain of the genus Mycobacterium is Mycobacterium abscessus, Mycobacterium abscessus subsp.abscessus, Mycobacterium abscessus subsp.massiliense, Mycobacterium abscessus subsp.bolletii, Mycobacterium smegmatis, Mycobacterium intracellulare, Mycobacterium avium, Mycobacterium kansasii, Mycobacterium fortuitum, Mycobacterium Mycobacterium gordonae, Mycobacterium osloensis, Mycobacterium phlei, Mycobacterium terrae, Mycobacterium chelonae, Mycobacterium mucogenicum, Mycobacterium peregrinum, Mycobacterium simiae, Mycobacterium wolinskyi, Mycobacterium paragordonae, Mycobacterium ulcerans, Mycobacterium marinum, Mycobacterium bovis, Mycobacterium bovis BCG,A pharmaceutical composition comprising at least one selected from the group consisting of Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium canetti, Mycobacterium caprae, Mycobacterium microti, Mycobacterium tuberculosisK strain, and Mycobacterium leprae.

9. In paragraph 6, A pharmaceutical composition, wherein the bacterial infection is at least one selected from the group consisting of nontuberculous mycobacterial infection, nontuberculous mycobacterial pulmonary disease, tuberculosis, leprosy, enterohemorrhagic Escherichia coli infection, enteropathogenic Escherichia coli infection, multidrug-resistant Pseudomonas aeruginosa infection, multidrug-resistant Acinetobacter baumannii infection, sepsis, pneumonia, enteritis, skin suppuration, bloodstream infection, wound infection, cystic fibrosis, mucositis, urinary tract infection, liver abscess, otitis media, keratitis, endophthalmitis, bacteremia, meningitis, cellulitis, and peritonitis.

10. A pharmaceutical composition according to claim 6, wherein the pharmaceutical composition further comprises an antibiotic.

11. In the 10th paragraph, the antibiotic is clarithromycin, polymyxin B, erythromycin, isoniazid, rifampicin, ethambutol, SQ-109, pyrazinamide, streptomycin, gentamicin, kanamycin, apramycin, capreomycin, ethionamide, prothionamide, enviomycin, para-aminosalicylic acid, cycloserine, amikacin, levofloxacin, Moxifloxacin, Gatifloxacin, ofloxacin, terizidone, thionamide, ethionamide, protionamide, clofazimine, linezolid, ampicillin, amoxicillin, clavulanate, thioacetazone, imipenem, cilastatin, bedaquiline, delamanid, rimipenem, cilastatin, meropenem, carbenicillin, chlorampenicol, tetracycline, doxycycline (doxycyline), tigecycline, minocycline,A pharmaceutical composition comprising at least one selected from the group consisting of and Collistin.

12. Health functional food for preventing or improving bacterial or fungal infections containing VLX600.

13. Antibacterial or antifungal over-the-counter drugs containing VLX600. 14.An antibacterial or antifungal cosmetic composition comprising VLX600. 15.An antibacterial or antifungal feed composition comprising VLX600.

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