Derivative compounds comprising an amino nicotinamide structure and antifungal pharmaceutical compositions comprising the same

Abstract

The present invention relates to derivative compounds comprising an amino nicotinamide structure, isomers thereof or pharmaceutically acceptable salts thereof, and antifungal pharmaceutical compositions comprising the same. More specifically, the present invention provides a benzyloxybenzylamine amino acid derivative with few side effects and an antifungal agent comprising the same, wherein the antifungal agent has antifungal activity against fungi selected from Candida, Sportricum, Malassezia, Cryptococcus, Aspergillus, Pneumocystis, and Mucor.

Description

Technology Field

[0001] The present invention relates to a derivative compound comprising an amino nicotinamide structure and a pharmaceutical composition for antifungal containing the same as an active ingredient.

[0002] The present invention was supported by the Ministry of Health and Welfare from August 1, 2022 to December 31, 2023 under the Infectious Disease Prevention and Treatment Technology Development Project, with the research project title "Optimization and Validation of Efficacy for Preclinical Treatment of Antifungal-Resistant Candida Infections," with Amtics Bio Co., Ltd. as the lead organization and the Korea Health Industry Development Institute as the research management agency. Background Technology

[0003] Fungal infections occur very frequently in patients whose immune function is compromised due to the excessive use of broad-spectrum antibacterial agents, organ transplantation, long-term administration of anticancer drugs, aging, and AIDS, or in patients whose immune function is compromised by procedures such as catheters or prosthetic devices.

[0004] Conventional azole-class drugs can lead to death from hepatitis when used in patients with impaired liver function, so liver function tests must be performed prior to administration. Flucytosine has been reported to cause dose-dependent bone marrow suppression and hepatotoxicity, as well as the potential for enterocolitis; since these side effects increase in patients with reduced renal function, monitoring the patient's renal function is crucial. It is also contraindicated in pregnant women. The primary toxicity of amphotericin B is glomerulonephropathy resulting from renal artery constriction; since this is dose-dependent, the incidence of permanent renal loss increases if the lifetime cumulative dose exceeds 4–5 g. Additionally, nephrotoxicity may occur due to tubular toxicity, including excessive loss of potassium, magnesium, and bicarbonate, as well as decreased erythropoietin production. Furthermore, acute reactions may include thrombophlebitis, chills, tremors, and hyperventilation. As such, existing antifungal agents exhibit various side effects depending on the type of drug, so there is a need for the development of new treatments that can enhance antifungal efficacy while reducing these side effects. The problem to be solved

[0005] One objective of the present invention is to provide a derivative compound comprising an amino nicodinamide structure, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof; and an antifungal composition comprising the same as an active ingredient that inhibits the formation of biofilms and / or the formation of antifungal-resistant strains of fungi.

[0006] However, the technical problems that the present invention aims to solve are not limited to those mentioned above, and other unmentioned problems will be clearly understood by those skilled in the art from the description below. means of solving the problem

[0007] Various embodiments of the present invention are described with reference to the drawings. In the following description, for a complete understanding of the present invention, various specific details, such as specific forms, compositions, and processes, are described. However, specific embodiments may be practiced without one or more of these specific details, or in combination with other known methods and forms. In other examples, known processes and manufacturing techniques are not described as specific details so as not to make the present invention unnecessary or obscure. Reference throughout this specification to one embodiment means that the specific features, forms, compositions, or characteristics described in association with the embodiment are included in one or more embodiments of the present invention. Accordingly, the circumstances of the embodiments expressed at various locations throughout this specification do not necessarily represent the same embodiment of the present invention. Additionally, specific features, forms, compositions, or characteristics may be combined in any suitable way in one or more embodiments. In the present invention, unless specifically noted otherwise, terms used have the general meaning known to those skilled in the art. In the present invention, all chemical formulas are intended to include any possible optical or geometric isomers (e.g., R form, S form, or racemic form, or cis-trans isomers of alkenes, etc.).

[0008] In one embodiment, a compound represented by the following formula (I), an isomer thereof, or a pharmaceutically acceptable salt is provided:

[0009] [Chemical Formula I]

[0010]

[0011] In the above formula, W is O or S; R1 is hydrogen or a C1-6 alkyl; R2 is -OX-R3; X is a C1-6 alkyl; R3 is a monocyclic or bicyclic heteroaryl group comprising one or two heteroatoms independently selected from nitrogen, oxygen, and sulfur, cycloalkyl groups having 5 to 8 carbon atoms, aryl groups, or nitrogen, in each case, these groups may be unsubstituted or substituted by one or two halogens or CF3; and Y is NH2, OH, OMe, or OEt. In the above embodiment, W is O; R1 is hydrogen; X is a C1-3 lower alkyl; R3 is a monocyclic heteroaryl group comprising a cycloalkyl group having 5 to 8 carbon atoms, a phenyl group, or nitrogen, in each case, these groups may be unsubstituted or substituted by one or two halogens or CF3; Y is NH2, providing a compound, an isomer thereof, or a pharmaceutically acceptable salt, wherein in the above embodiments, W is O; R1 is hydrogen; X is a methyl group; and R3 is a cycloalkyl group having 5 to 8 carbon atoms, a phenyl group, or a pyridyl group, in each case, these groups may be unsubstituted or substituted by one or two halogens or CF3; Y is NH2, providing a compound, an isomer thereof, or a pharmaceutically acceptable salt.

[0012] In one embodiment, a pharmaceutical composition for the prevention or treatment of fungal infections is provided, comprising the compound, its isomer, or a pharmaceutically acceptable salt as an active ingredient. In the above embodiment, a pharmaceutical composition is provided having antibacterial activity against bacteria selected from Candida, Sportricum, Malassezia, Cryptococcus, Aspergillus, Pneumocystis, and Mucor; and in the above embodiment, the mycosis is meningitis, encephalitis caused by meningitis, cutaneous candidiasis, mucosal candidiasis, visceral candidiasis, urinary tract candidiasis, candidal endocarditis, oropharyngeal candidiasis, candidal endophthalmitis, candidal sepsis, ringworm, tinea versicolor, sportrixosis, fungal abscess, fungal granuloma, pyogenic granuloma, Madura mycosis, Pneumocystis pneumonia, systemic cryptococcosis, cutaneous and mucosal cryptococcosis, aspergillosis, cavity, hemoptysis, allergy, chronic pulmonary tuberculosis. A pharmaceutical composition is provided that is selected from pulmonary fibrosis, pulmonary cysts, chronic fever, cough, sputum, hemoptysis, ringworm, thrush, fungal dementia, and zygomycosis.

[0013] In the present invention, the halogen is F, Cl, Br, or I.

[0014] In the present invention, the term “C 1-7 “Alkyl” means a straight-chain or branched-chain alkyl having 1 to 7 carbon atoms, and is not limited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl, etc.; preferably includes ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, and tert-butyl.

[0015] In the present invention, the term “cycloalkyl” means a cyclic alkyl having three or more carbon atoms in a ring, and includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and cyclodecyl. The terms “C3-C8 cycloalkyl”, “C3-C7 cycloalkyl”, and “C3-C6 cycloalkyl” have similar meanings.

[0016] In the present invention, the terms “aromatic heterocyclic” or “heteroaryl” have the same meaning and refer to a heteroaromatic group comprising one or more heteroatoms. The heteroatoms referred to herein include oxygen, sulfur, and nitrogen. Examples include furyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl ring may be condensed to an aryl, heterocyclic group, or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring. The heteroaryl is optionally substituted or unsubstituted.

[0017] In the present invention, the term “substitution” means that one or more hydrogen atoms on a specified group are substituted by a specified substituent. The specified substituent is the substituent correspondingly described above or the substituent shown in each example. Unless specifically described, the specified substituted group may have one substituent selected from a specific group at any substitutable site of the group, and said substituent may be the same or different at each position. Cyclic substituents, such as heterocycloalkyl, may be connected to another ring, such as cycloalkyl, to form a spiro-double ring system, such as a two-ring system having one common carbon atom. Those skilled in the art should understand that the combination of substituents anticipated in the present invention is a stable or chemically feasible combination. The said substituents are, for example, C 1-8 Alkyl, C 2-8 Alkenyl, C 2-8 Alkinyl, C 3-8 Cycloalkyl, 3- to 12-membered heterocyclic group, aryl, heteroaryl, halogen, hydroxyl, carboxyl(-COOH), C 1-8 Aldehyde group, C 2-10 Asil, C 2-10 Ester group, C 1-C12 Alkoxycarbonyl, amino, alkoxy, C 1-10Sulfonyl, etc., but are not limited to these.

[0018] In one embodiment of the present invention, a compound represented by the following formula 1, an isomer thereof, or a pharmaceutically acceptable salt is provided:

[0019] [Chemical Formula 1]

[0020]

[0021] In the above formula, W is O or S; R1 is hydrogen or a C1-6 alkyl; R2 is -OX-R3; X is a C1-6 alkyl; and R3 is a cycloalkyl group having 5 to 8 carbon atoms, an aryl group, or a monocyclic or dicyclic heteroaryl group comprising one or two heteroatoms independently selected from nitrogen, oxygen, and sulfur, wherein in each case these groups are unsubstituted or one or two halogens, CF 3, It can be substituted by an ethoxy group; Y is substituted by one or more amino groups, and is a heteroaryl or C1 to C comprising one or two heteroatoms independently selected from nitrogen, oxygen, and sulfur. 10 It is an alkylamino group.

[0022] In this specification, the term "alkylamino" refers to a group having the structure-NHR', where R' is hydrogen, alkyl or substituted alkyl, cycloalkyl or substituted cycloalkyl as defined above.

[0023] In this specification, the term “dialkylamino” refers to a group having the structure-NRR’, wherein R and R’ are each independently an alkyl or substituted alkyl, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted cycloalkenyl, aryl or substituted aryl, heterocyclil or substituted heterocyclil as defined above. R and R’ may have the same or different dialkylamino moiety. Examples of dialkylamino groups include, but are not limited to, dimethylamino, methyl ethylamino, diethylamino, methylpropylamino, di(n-propyl)amino, di(iso-propyl)amino, di(cyclopropyl)amino, di(n-butyl)amino, di(tert-butyl)amino, di(neopentyl)amino, di(n-pentyl)amino, di(hexyl)amino, di(cyclohexyl)amino, etc. In certain embodiments, R and R’ are connected to form a cyclic structure. The resulting cyclic structure may be aromatic or non-aromatic. Examples of cyclic diaminoalkyl groups include, but are not limited to, aziridinyl, pyrrolidinyl, piperidinyl, morpholinyl, pyrrolyl, imidazolyl, 1,3,4-trianolyl, and tetrazolyl.

[0024] In another embodiment of the present invention, W is O; R1 is hydrogen; X is a C1-3 lower alkyl; and R3 is a cycloalkyl group having 5 to 8 carbon atoms, a phenyl group, or a monocyclic heteroaryl group containing nitrogen, wherein in each case these groups are unsubstituted or have one or two halogens, CF 3, It can be substituted by a methoxy group; Y is substituted by one amino group, and is a heteroaryl or C1 to C comprising one or two nitrogen heteroatoms. 10 Provides a compound, its isomer, or a pharmaceutically acceptable salt that is an alkylamino group of

[0025] In another embodiment of the present invention, W is O; R1 is hydrogen; X is a methyl group; and R3 is a cycloalkyl group having 5 to 8 carbon atoms, a phenyl group, or a pyridyl group, wherein in each case these groups are unsubstituted or have one or two halogens, CF 3, Provides a compound, isomer thereof, or pharmaceutically acceptable salt that can be substituted by a methoxy group; Y is substituted by one amino group and is a heteroaryl or C1 to C6 alkylamino group comprising one nitrogen heteroatom.

[0026] The pharmaceutical composition of the present invention comprises the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier within a safe effective amount range. Herein, “safe effective amount” refers to an amount of the compound capable of significantly improving a disease without causing serious side effects. Typically, the pharmaceutical composition contains 1 to 2000 mg of the compound / agent of the present invention, and more preferably 5 to 500 mg of the compound / agent of the present invention. Preferably, the “agent” is a single capsule or tablet.

[0027] “Pharmaceuticalally acceptable carriers” means one or more compatible solid or liquid fillers or gel materials that are suitable for human use and have sufficient purity and sufficiently low toxicity. Here, “compatibility” means that each component in the composition and the compound of the present invention are mixed together without significantly reducing the efficacy of the compound. Some examples of pharmaceutically acceptable carriers include cellulose and derivatives thereof (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g., stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g., Tween®), humectants (e.g., sodium dodecyl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, and pyrogen-free water.

[0028] As used herein, the terms "optical isomer" or "stereoisomer" refer to any of the various stereoisomer coordinations that may exist for a given compound of the present invention, including geometric isomers. Substituents are understood to be capable of attaching to the chiral center of a carbon atom. The term "chiral" refers to a molecule having the characteristic of non-superimposability on its mirror image partner, whereas the term "achiral" refers to a molecule capable of superposition on its mirror image partner. Accordingly, the present invention includes enantiomers, diastereomers, or racemics of a compound. "Enantiomers" are a pair of stereoisomers that are mirror images of each other and are non-superimposable. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. The term is used to designate racemic mixtures where appropriate. "Diastereomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. Absolute stereochemistry is specified according to the Cahn-Ingold-Prelog RS system. If the compound is a pure enantiomer, the stereochemistry at each chiral carbon may be specified by R or S. Resolved compounds whose absolute coordination is unknown may be designated as (+) or (-) depending on the direction in which they rotate plane polarization at the wavelength of the sodium D line (left-handed or right-handed). The specific compounds described herein may contain one or more asymmetric centers or axes and thus produce enantiomers, diastereomers, and other stereoisomer forms that can be defined as (R)- or (S)- in terms of absolute stereochemistry.

[0029] The method of administration of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative methods of administration include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular, or subcutaneous), and local administration.

[0030] Solid formulations for oral administration include capsules, tablets, pills, powders, and granules. In these solid formulations, the active compound is mixed with at least one common inert excipient (or carrier), for example, sodium citrate or dical phosphate, or with the following components: (a) fillers or compatibilizers such as starch, lactose, sucrose, glucose, mannitol, and silica; (b) adhesives such as hydroxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia gum; (c) humectants such as glycerin; (d) disintegrants such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, some complex silicates, and sodium carbonate; (e) retardants such as paraffin; (f) absorption promoters such as quaternary amine compounds; (g) wetting agents such as cetanol and glyceryl monostearate; (h) an adsorbent such as kaolin; and (i) a lubricant such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or a mixture thereof. In capsules, tablets, and pills, the formulation may include a buffer.

[0031] Solid formulations such as tablets, coated tablets, capsules, pills, and granules may be manufactured using coating and shell materials, such as enteric coatings and other materials known in the art. These may include an opacifying agent, and the release of the active compound or compounds in these compositions may be released in a delayed manner in any part of the digestive tract. Examples of usable embedding components are polymeric materials and wax-based materials. If necessary, the active compound may form a microcapsule with one or more of the excipients.

[0032] Liquid formulations for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, the liquid formulation may include an inert diluent commonly used in the art, such as water or other solvents, solubilizers, and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0033] In addition to these inert diluents, the composition may include auxiliary agents such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and flavorings.

[0034] In addition to the active compound, the suspension may include a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan, microcrystalline cellulose, aluminum methoxide and agar, or a mixture of these materials.

[0035] Compositions for parenteral injection may comprise physiologically acceptable sterile water or anhydrous solution, dispersion, suspension or emulsion, and sterile powder that is redissolved into a sterile injectable solution or dispersion. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0036] Formulations of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalants. Under sterile conditions, the active ingredient is mixed with a physiologically acceptable carrier and any preservative, buffer, or propellant that may be required.

[0037] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds. In some preferred embodiments, the compounds of the present invention may be administered by forming a PROTAC with other small molecule compounds or by forming an ADC with other macromolecular compounds, such as monoclonal antibodies.

[0038] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (e.g., human) requiring treatment, wherein the dose administered is a pharmaceutically recognized effective dose, and for a human weighing 60 kg, the daily dose is typically 1 to 2000 mg, preferably 5 to 500 mg. Of course, the specific dose must take into account factors such as the route of administration and the patient's health condition, all of which are within the scope of a skilled physician's expertise.

[0039] In one embodiment, fungi are a group of eucalyptus organisms having cell walls, including but not limited to yeast, mold, mushrooms, etc. The cell walls of fungi contain chitin and beta-glucan, which are characteristic components, and thus are targets for the development of antifungal agents targeting fungi. In addition, ergosterol, a component of the fungal cell membrane, is a major target. For example, Azoles inhibit the synthesis of ergosterol and destroy the cell membrane. In addition, drugs such as flucytosine inhibit the proliferation of fungal cells by interfering with DNA and RNA synthesis, some antifungal agents block fungal protein synthesis by targeting ribosomes, and drugs such as griseofulvin inhibit cell division by interfering with microtubule function.

[0040] In the present invention, "prevention" may include, without limitation, any act of blocking disease symptoms or suppressing or delaying disease symptoms using the pharmaceutical composition of the present invention.

[0041] In the present invention, "treatment" and "improvement" may be included without limitation as long as they are any act of improving or benefiting from disease symptoms by irradiating the pharmaceutical composition of the present invention.

[0042] The "pharmaceutical composition" of the present invention is not limited to these, but may be formulated and used in the form of oral formulations such as powders, granules, capsules, tablets, and aqueous suspensions, as well as topical preparations, suppositories, and sterile injectable solutions, according to conventional methods. The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. For oral administration, the pharmaceutically acceptable carrier may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, colorants, flavorings, etc. For injectable preparations, it may include buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers, etc., in combination; and for topical administration, a base, excipients, lubricants, preservatives, etc. may be used. The formulations of the pharmaceutical composition of the present invention may be prepared in various ways by mixing with the pharmaceutically acceptable carriers described above. For example, for oral administration, it can be manufactured in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and for injectables, it can be manufactured in the form of unit dosing ampoules or multiple dosing ampoules. In addition, it can be formulated as a solution, suspension, tablet, capsule, sustained-release formulation, etc. Meanwhile, examples of carriers, excipients, and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil. In addition, it may additionally include fillers, anticoagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, etc.

[0043] In the present invention, "routes of administration" for a pharmaceutical composition include, but are not limited to, oral, intravenous, intramuscular, intra-arterial, intramedullary, intradural, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual, or rectal. Oral or parenteral administration is preferred.

[0044] In the present invention, "parenteral" includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intradural, intralesional, and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may also be administered in the form of a suppository for rectal administration.

[0045] The "use and dosage" of the composition of the present invention may vary depending on several factors including the activity of the specific compound used, age, body weight, general health, gender, diet, time of administration, route of administration, elimination rate, drug combination, and the severity of the specific disease to be prevented or treated, and the dosage of the pharmaceutical composition may be appropriately selected by a person skilled in the art, depending on the patient's condition, body weight, degree of disease, form of medication, route of administration, and duration, and may be administered at a dose of 0.0001 to 50 mg / kg or 0.001 to 50 mg / kg per day. Administration may be administered once a day or divided into several doses. The dosage does not limit the scope of the present invention in any way. The pharmaceutical composition according to the present invention may be formulated as a pill, coated tablet, capsule, liquid, gel, syrup, slurry, or suspension.

[0046] In one embodiment of the present invention, a compound represented by the following chemical formula 1, an isomer thereof, or a pharmaceutically acceptable salt is provided:

[0047] [Chemical Formula 1]

[0048]

[0049] In the above formula, W is O or S; R1 is hydrogen or a C1-6 alkyl; R2 is -OX-R3; X is a C1-6 alkyl; and R3 is a cycloalkyl group having 5 to 8 carbon atoms, an aryl group, or a monocyclic or dicyclic heteroaryl group comprising one or two heteroatoms independently selected from nitrogen, oxygen, and sulfur, wherein in each case these groups are unsubstituted or one or two halogens, CF 3, It can be substituted by an ethoxy group; Y is substituted by one or more amino groups, and is a heteroaryl or C1 to C comprising one or two heteroatoms independently selected from nitrogen, oxygen, and sulfur. 10 It is an alkylamino group.

[0050] In another embodiment of the present invention, W is O; R1 is hydrogen; X is a C1-3 lower alkyl; and R3 is a cycloalkyl group having 5 to 8 carbon atoms, a phenyl group, or a monocyclic heteroaryl group containing nitrogen, wherein in each case these groups are unsubstituted or have one or two halogens, CF 3, It can be substituted by a methoxy group; Y is substituted by one amino group, and is a heteroaryl or C1 to C comprising one or two nitrogen heteroatoms. 10 Provides a compound, its isomer, or a pharmaceutically acceptable salt that is an alkylamino group of

[0051] In another embodiment of the present invention, W is O; R1 is hydrogen; X is a methyl group; and R3 is a cycloalkyl group having 5 to 8 carbon atoms, a phenyl group, or a pyridyl group, wherein in each case these groups are unsubstituted or have one or two halogens, CF 3, Provides a compound, isomer thereof, or pharmaceutically acceptable salt that can be substituted by a methoxy group; Y is substituted by one amino group and is a heteroaryl or C1 to C6 alkylamino group comprising one nitrogen heteroatom.

[0052] In another embodiment of the present invention, the compound represented by Formula 1 provides a compound selected from the group consisting of compounds represented by Formulas 2 to 20 below, an isomer thereof, or a pharmaceutically acceptable salt.

[0053] [Chemical Formula 2]

[0054]

[0055] [Chemical Formula 3]

[0056]

[0057] [Chemical Formula 4]

[0058]

[0059] [Chemical Formula 5]

[0060]

[0061] [Chemical Formula 6]

[0062]

[0063] [Chemical Formula 7]

[0064]

[0065] [Chemical Formula 8]

[0066]

[0067] [Chemical Formula 9]

[0068]

[0069] [Chemical Formula 10]

[0070]

[0071] [Chemical Formula 11]

[0072]

[0073] [Chemical Formula 12]

[0074]

[0075] [Chemical Formula 13]

[0076]

[0077] [Chemical Formula 14]

[0078]

[0079] [Chemical Formula 15]

[0080]

[0081] [Chemical Formula 16]

[0082]

[0083] [Chemical Formula 17]

[0084]

[0085] [Chemical Formula 18]

[0086]

[0087] [Chemical Formula 19]

[0088]

[0089] [Chemical Formula 20]

[0090]

[0091] In another embodiment of the present invention, a pharmaceutical composition for the prevention or treatment of fungal infections is provided, comprising the compound, its isomer, or a pharmaceutically acceptable salt as an active ingredient.

[0092] In another embodiment of the present invention, the pharmaceutical composition is provided having antibacterial activity against bacteria selected from Candida, Sportricum, Malassezia, Cryptococcus, Aspergillus, Pneumocystis, and Mucor.

[0093] In another embodiment of the present invention, a pharmaceutical composition is provided in which the mycosis is selected from meningitis, encephalitis caused by meningitis, cutaneous candidiasis, mucosal candidiasis, visceral candidiasis, urinary tract candidiasis, candidal endocarditis, oropharyngeal candidiasis, candidal endophthalmitis, candidal sepsis, ringworm, tinea versicolor, sportrixosis, fungal abscess, fungal granuloma, pyogenic granuloma, Madura mycosis, Pneumocystis pneumonia, systemic cryptococcosis, cutaneous and mucosal cryptococcosis, aspergillosis, cavities, hemoptysis, allergy, chronic pulmonary tuberculosis, pulmonary fibrosis, pulmonary cysts, chronic fever, cough, sputum, hemoptysis, ringworm, thrush, fungal dementia, and zygomycosis. Effects of the invention

[0094] The compound of the present invention has a high antifungal effect while having low side effects. Brief explanation of the drawing

[0095] Figure 1 shows fluconazole (FCZ) and ATB6015. in vivo The efficacy was compared and analyzed. Figure 2 is Candida glabrata This is a measurement of the frequency of appearance of resistant strains in [location]. Fig. 3 is Candida albicans (12-99) Changes in body weight and survival rate after administration of ATC0001 free in an infection model. Figure 4 is the result of a cross-check commissioned to another specialized agency, Candida albicans (12-99) This concerns changes in body weight and survival rate after administration of ATC0001 free in an infection model. Fig. 5 is Candida auris This is the change in body weight and survival rate after drug treatment in the infection model. Figure 6 shows the five additionally synthesized compounds (ATC0013, ATC0017, ATC0018, ATC0019, ATC0020). in vitro This is the result of the active screening. Figure 7a shows the spectroscopic data of ATB6015. Figure 7b shows the spectroscopic data of ATC0001. Figure 7c is the spectroscopic data of ATC0002. Figure 7d shows the spectroscopic data of ATC0003. Figure 7e shows the spectroscopic data of ATC0004. Figure 7f shows the spectroscopic data of ATC0005. Figure 7g shows the spectroscopic data of ATC0006. Figure 7h shows the spectroscopic data of ATC0007. Figure 7i shows the spectroscopic data of ATB6015 chloride. Figure 7j is spectroscopic data of ATC0001 chloride. Figure 7k shows the spectroscopic data of ATC0002 chloride. Figure 71 shows the spectroscopic data of ATC0003 chloride. Figure 7m shows the spectroscopic data of ATC0004 chloride. Figure 7n shows the spectroscopic data of ATC0005 chloride. Figure 70 shows the spectroscopic data of ATC0013. Figure 7p shows the spectroscopic data of ATC0017. Figure 7q is the spectroscopic data of ATC0018. Figure 7r is the spectroscopic data of ATC0019. Figure 7s is the spectroscopic data of ATC0020. Specific details for implementing the invention

[0096] The present invention will be described in more detail below through examples. These examples are intended solely to explain the present invention more specifically, and it will be obvious to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the invention.

[0097] Example 1. C. auris ATB6015 using an infection animal model in vivo Drug efficacy evaluation and optimization

[0098] Example 1-1. ATB1651-based derivative ATB6015 Candida Regarding the 3 types in vitro Confirmation of antifungal efficacy

[0099] The inventors prepared a new derivative of the following chemical formula 2 (ATB6015) based on the structure-activity relationship (SAR) of a conventional amino acid derivative (ATB1651) having excellent pharmacological and antifungal activity:

[0100] [Chemical Formula 2]

[0101]

[0102] The above derivative includes antifungal resistance. Candida 3 types ( C. albicans wild type strain SC5314, azole antifungal resistant strain C. glabrata BG2 strain, a multidrug-resistant fungus C. auris Regarding the B8441 strain in vitro As shown in Table 1 below, it was confirmed that it exhibited 4 to 25 times better antifungal activity under the conditions.

[0103] ATB6015, a derivative of ATB1651 Candida Antimicrobial activity against 3 types Drug MIC (µg / mL) C. albicans (SC5314) C. glabrata (BG-2) C. auris (B8441) Amphotericin B 0.25 0.5 0.5 Fluconazole 1 >64 >64 ATB1651 4 8 8 ATB6015 0.25 2 0.5

[0104] Example 1-2. Multidrug-resistant bacteria Candida auris Construction of an infected mouse model and ATB6015 in vivo Confirmation of antifungal efficacy

[0105] Evaluation of the drug efficacy of ATB6015 for systemic infection C. auris To establish an infection animal model, 7-week-old female A / J mice C. auris The number of cells is 1x10 respectively 7 , 5x10 7 , 1x10 8 , 2x10 8 Survival was checked after infecting via intravenous administration, adjusted to (cells). The number of infected cells was 1x10 7Except for the case where all mice died at 2 days after infection, the number of infected cells was set to 1x10 7 It was selected as.

[0106] Next, Candida auris To confirm the therapeutic efficacy of ATB6015 on mice infected with [variable], the previously established cell concentration (1x10⁻¹⁰) 7 Align the mouse with ) C. auris Infecting and the previously known antifungal agents fluconazole (FCZ) and ATB6015 in vivo The efficacy was compared and analyzed (see Figure 1). After infection, ATB6015 was administered orally five times at a concentration of 10 mpk (at 3, 24, 48, 60, and 72 hours after infection). As a result, the median survival days of the untreated group (vehicle) and the fluconazole-treated group were 6 days and 11.5 days, respectively, while the median survival days of the ATB6015-treated group increased significantly to 20 days.

[0107] Examples 1-3. Drug Target Discovery of ATB6015 Using HIP Assay (Haploinsufficiency Profiling Assay)

[0108] As a result of performing a HIP assay after treatment with ATB6015, we were able to identify drug target candidates with significantly reduced competitive fitness and low p-values. Among these, GWT1, which had the lowest p-value and a log2 fold change of -2 or less, was identified as the most likely drug target for ATB6015, as it is known to be a gene involved in the synthesis of glycosyl-phosphatidylinositol (GPI), which is related to cell wall integrity (Watanabe et al., 2012, Antimicrob Agents Chemother).

[0109] Example 2. Evaluation of Physical Properties of ATB6015

[0110] Example 2-1. Evaluation of the Solubility of ATB6015

[0111] Since drug solubility is important for toxicity tests for oral (per oral, PO) and intravenous injection (IV) administration, as well as for the future development of the drug as an injectable, the solubility of the antifungal candidate ATB6015 was evaluated by dissolving ATB6015 in various excipient components used as excipients in animal experiments and assessing its solubility in each solvent. The solubility score was rated on a 6-point scale, ranging from 1 for insolubility to 6 for complete dissolution. It was rated as 2 for particles remaining and settling or layer separation occurring, 3 for particles remaining but not settling, 4 for fine particles being visible, and 5 for cloudy complete dissolution or a long dissolution time.

[0112] For intravenous toxicity tests, when ATB6015 was dissolved at a level of 6 mg / ml in various excipients such as PEG, Tween, and Cyclodextrin (CD), as shown in Tables 2 and 3 below, ATB6015 did not dissolve in water, and particles remained in other excipients, settled, or separated into layers (Table 2). In addition, when dissolved at a high concentration of 100 mg / ml for single oral toxicity tests, it hardly dissolved, so it was determined that it is desirable to increase the drug's activity or solubility (Table 3).

[0113] Evaluation of the Solubility of ATB6015 in Various Excipients Brothers Degree of dissolution * 0.9% saline 2 1X PBS 2 DW 1 *0.05% MC in DW 2 10% PEG400 + 90% DW 2 30% PEG400 + 70% DW 2 10% PEG300 + 90% DW 2 20% PEG300 + 80% DW 2 30% PEG300 + 70% DW 2 5% Tween80 + 95% DW 2 ATB6015 : CD = 1:1 (w / w) mix + DW 2 (ATB6015 : CD = 1:1(w / w)) : (10% PEG300 + 90% DW) = 1:1 2 5% PEG400 + 95% DW 2 5% PEG400 + 95% Saline 2 10% PEG400 + 90% Saline 2 100mg / kg Glycerol in DW 3 40% CD in DW 4

[0114] Brothers Degree of dissolution * 10% (50% Castor oil + 50% EtOH) + 90% DW 1 50% (50% Castor oil + 50% EtOH) + 50% DW 1 50% PEG400 + 50% DW 1 50% propylene glycol + 50% DW 1 1% Tween80 + 99% DW 3 5% ethanol + 95% DW 1 1% DMSO + 99% DW 2

[0115] Example 2-2. Measurement of Physicochemical Parameters of ATB6015

[0116] Since the physicochemical parameters of a new drug candidate are essential for assessing future PK characteristics and optimizing the candidate, the most important physical properties (physichochemiocal parameters) of ATB6015—solubility, pKa, lipophilicity, and melting temperature—were analyzed.

[0117] (1) Solubility of a drug is one of the most important physical properties in new drug development. As shown in the table below, the intrinsic solubility of ATB6015 was determined by the pH-metric method (potentiometric method) in the pH range from 2 to 12, and the result was 94.19 μM as shown in Table 4 below.

[0118] Intrinsic Sol. T / ℃ Method Ionic Environment Assay Type 94.18 μM 24.5 pH-metric 0.167 M KCl Curve fitting

[0119] (2) The degree of ionization (pKa) is a property that can predict the physicochemical behavior of a compound and is a parameter that affects lipidity, solubility, protein binding, and permeability. As shown in Table 5 below, the pKa of ATB6015 was found to have two values ​​as structural characteristics of the compound, with values ​​of 3.89 and 5.17.

[0120] pKa Type T / ℃ Ionic Environment Method 3.89 ± 0.05 Base 24.9 0.171 M KCl pH-metric 5.17 ± 0.03 Base 24.9 0.171 M KCl pH-metric

[0121] (3) Lipophilicity (logP / logD) is a physical property that is very related to the absorption of a drug in the human body. It was analyzed by measuring the partition coefficient in water and octanol, and as shown in Table 6 below, the LogP value of ATB6015 was measured to be 2.41.

[0122] LogP Type LogD at pH 7.4 (blood) T / ℃ Ionic Environment Method 2.41 ± 0.03 Neutral 2.40 25.0 0.169 M KCl pH-metric

[0123] (4) The melting temperature is an intrinsic property of the compound. As a result of measuring the melting temperature using differential scanning calorimetry (DSC), values ​​of 163°C to 165°C were confirmed as shown in Table 7 below.

[0124] melting point Analysis method Analysis conditions Sampling / Preprocessing 163℃ - 165℃ DSC Analysis -10℃ ~ 300℃ range 52 min Standard Sealing Pan(GCA-0052, Aluminum)

[0125] Example 3. Development of Candidate Materials for Optimization of Physical Properties Improvement of ATB6015

[0126] Example 3-1. Synthesis of ATB6015, 5 derivatives, and 6 chlorides (salt form)

[0127] To find a substance with improved antifungal activity and solubility compared to the antifungal candidate ATB6015, seven derivatives of ATB6015 and five salt-form compounds thereof were synthesized.

[0128] Synthesis method of antifungal candidate ATB6015 and 7 derivatives (ATC0001 - ATC0007)

[0129] Compound 1 was synthesized as follows after Boc protection using Boc anhydride, NaHCO3, and MeOH solvent as starting materials and stirring at room temperature for 24 hours.

[0130]

[0131] Intermediates 3a-3h with functional groups were synthesized as follows by refluxing substances 2a-2h having functional groups in compound 1 with K2CO3 and acetone solvent.

[0132]

[0133] Salt compounds 4a-4h were synthesized as follows by deprotecting intermediates 3a-3h with introduced functional groups by stirring at room temperature for 24 hours using 4.0 M HCl in dioxane and EtOAc solvents.

[0134]

[0135] Compound 6 (ATB6015) and seven derivatives (ATC0001 - ATC0007, compound numbers 6-13) were finally synthesized at room temperature using DMTMM and DMF solvents with substance 5 having a functional group and different analogues 4a - 4h.

[0136]

[0137] Synthesis method of chlorides (salt form) of ATB6015 and its five derivatives (ATC0001 - ATC0005)

[0138] Compounds 14 (ATB6015 salt) and compounds 15-19 (5 derivatives, ATC0001 salt - ATC0005 salt) were finally synthesized at room temperature using 4.0 M dioxane and EtOAc solvents in compounds 6-11 synthesized above as follows.

[0139]

[0140] Through the above compound synthesis method, the antifungal candidate ATB6015 and 7 derivatives (ATC0001-0005) and 6 chlorides (salt forms) were successfully synthesized, and the drug names are as follows. In addition, NMR data are shown in Figure 7a and below.

[0141] turn Drug names structure 1 ATB6015 2 ATC0001 free 3 ATC0002 free 4 ATC0003 free 5 ATC0004 free 6 ATC0005 free 7 ATC0006 free 8 ATC0007 free 9 ATB6015 salt 10 ATC0001 salt 11 ATC0002 salt 12 ATC0003 salt 13 ATC0004 salt 14 ATC0005 salt

[0142] Example 3-2. Synthesis of 5 additional ATB6015 derivatives (ATC0013, ATC0017, ATC0018, ATC0019, ATC0020)

[0143] Compound 20 was synthesized as follows by using NMM, IBCF, and THF solvents in the starting material and 4b, and stirring at room temperature for 2 hours.

[0144]

[0145] Compound 21 (ATC0013) was finally synthesized at room temperature using 4.0 M in dioxane and EtOAc solvent in compound 20 synthesized above as follows.

[0146]

[0147] Compound 23 was synthesized at room temperature using DMTMM and DMF solvents on materials 5 and 22 having functional groups as follows.

[0148]

[0149] Different analogues 24a–24d of compound 23 synthesized above were prepared using CS2CO3 and DMF solvents as follows, 120 oFour derivatives 25a-25d were finally synthesized using a microwave reactor in C.

[0150]

[0151] Through the above compound synthesis method, five additional derivatives (ATC0013, ATC0017-0020) were successfully synthesized, and their names are as follows. In addition, NMR data are shown in Figure 7a and below.

[0152] turn Drug names structure 1 ATC0013 2 ATC0017 3 ATC0018 4 ATC0019 5 ATC0020

[0153] Example 3-3. For deriving optimized candidate substances based on ATB6015 in vitro Active screening

[0154] By analyzing the Minimal Inhibitory Concentration (MIC) using the 13 candidate drugs of the above derivatives synthesized based on ATB6015, drug resistance White To screen for the ability to inhibit strains and invasive strains, drug activity evaluations were performed on the following fungi:

[0155] 4 types of invasive fungi ( Candida albicans (SC5314) , Candida tropicalis (KCCM 50075), Fusarium oxysporum (KCCM60555) and Aspergillus fumigatus (CEA10))

[0156] - Multidrug resistance showing resistance to antifungal agents White strain ( Candida albicans (12-99) , Candida glabrata (BG-2))

[0157] The test drug was dissolved in 100% DMSO solvent and treated in the medium at a final ratio of 1% in accordance with the Clinical Standards Institute (CLSI) Guidelines Edition 4 M27 (Table 2-1). C. albicans (SC5314), C. albicans (12-99), C. glabrata (BG-2) and Candida tropicalisThe final number of cells for (KCCM 50075) was prepared as 500 cells, and the experiment was conducted as shown in Table 10 below.

[0158] RPMI 1640 medium 10.4 grams MOPS buffer 34.53 grams NaOH Adjust the pH to 7 the whole volume 1 L가 아니다 마근증류수 그리

[0159] MIC analysis results showed that among the ATB6015 derivatives, ATC0001 free was effective against two types of invasive pathogens and two types of azole-resistant bacteria White Overall, results showed lower MIC values ​​compared to ATB6015 for spp. ATC0001 free exhibited the greatest increase in antifungal activity relative to ATB6015 among the 13 derivatives, and various White It showed efficacy against spp., particularly azole-resistant strains C. albicans (12-99) and C. glabrata ATC0001 free for (BG2) in vitro As shown in Table 11 below, it was confirmed that the antifungal activity increased by approximately 10 to 32 times compared to ATB6015. In addition, the ATC0003 free, ATC0003 salt, and ATC0006 free compounds C. albicans (SC5314), C. albicans (12-99) and C. tropicalis At a level similar to ATB6015 in the (KCCM 50075) strain in vitro It exhibited antifungal activity, and ATC0004 free is an azole-resistant strain C. albicans (12-99) and C. glabrata (BG2) species compared to ATB6015 in vitro The antifungal efficacy improved by about 2 to 4 times.

[0160] Drug MIC (㎍ / mL) C. albicans (SC5314) C. albicans (12-99) C. glabrata (BG-2) C. tropicalis (KCCM 50075) Amphotericin B 0.25 0.25 0.5 0.125 Fluconazole 1 >64 >64 >64 ATB6015 0.25 4 2 1-4 ATC0001 free 0.03 0.125 0.25 0.5-1 ATC0002 free 8 64 64 64 ATC0003 free 0.5 4 8 4 ATC0004 free 0.25 1 1 >64 ATC0005 free >64 >64 >64 >64 ATC0006 free 1 4 4 2 ATC0007 free >64 >64 >64 >64 ATB6015 salt 0.5 8 2 2 ATC0001 salt 0.03 0.125 0.25 >64 ATC0002 salt 8 64 64 4 ATC0003 salt 0.5 4 8 1 ATC0004 salt 0.25 >64 >64 >64 ATC0005 salt >64 >64 >64 4

[0161] Meanwhile, the above compound is White not only spp. Aspergillus spp. and Fusarium It also exhibited antifungal activity against filamentous fungi such as spp. Aspergillus fumigatus (CEA10) and Fusarium oxysporumMIC measurement results from (KCCM60555), as shown in Table 12 below Aspergillus fumigatus For the strain, the antifungal activity of ATC0001 free was improved by approximately 17 times, and Fusarium oxysporum Regarding this, the antifungal activity of ATC0001-free was 66 times higher. In addition, azole-resistant strains C. albicans (12-99) and C. glabrata In the case of ATC0004 free, which showed 2 to 4 times improved antifungal efficacy compared to ATB6015 in (BG2), Fusarium oxysporum (KCCM60555) did not show efficacy, but Aspergillus fumigatus (CEA10) showed an effect that was improved by about 4 times.

[0162] compound A. fumigatus (CEA10) F.oxysporum (KCCM60555) ATB6015 0.5∼1 2 ATC0001 free 0.03∼0.06 0.03 ATC0002 free 4∼8 4∼8 ATC0004 free 0.125∼0.5 >64 ATC0005 free >64 >64

[0163] Summarizing the MIC screening results of ATB6015-based derivatives, ATC0001-free is invasive White Fungi, multidrug resistance White and also in filamentous fungi in vitro It was confirmed that the compound possesses antifungal activity and exhibits a broad spectrum of effects against various strains.

[0164] Example 3-4. addition of the synthesized derivative in vitro active

[0165] Five additional compounds containing amino nicotinamide structure derivatives (ATC0013, ATC0017, ATC0018, ATC0019, ATC0020) in vitro Active screening was performed.

[0166] Drug activity evaluations were performed on the following fungi to screen whether five candidate drugs could inhibit representative Candida strains by analyzing the Minimal Inhibitory Concentration (MIC):

[0167] - 2 species of invasive fungi (Candida albicans (SC5314), Candida auris (B8441))

[0168] The test was performed according to the Minimum Inhibitory Concentration (MIC) test developed by the European Committee on Antimicrobial Susceptibility Testing (EUCAST). The test drug was dissolved in 100% DMSO solvent and added to the medium at a final concentration of 2%, using a cell concentration corresponding to an OD600 of 0.5 for C. albicans (SC5314) and C. auris (B8441). 200 μl of the cell suspension was mixed with 10 ml of RPMI 1640 medium (pH 7.4, 0.165 M MOPS, and 2% glucose) and dispensed into 96-well plates to contain the diluted drug. The 96-well plates were incubated at 35°C for 2 days, and the cell concentration in each well was measured at an OD595. After measurement, the culture solution was plated onto YPD medium and incubated at 30°C for 24 hours to confirm the cidal effect.

[0169] As a result of MIC analysis, as shown in Figures 6a and 6b, it was confirmed that among the five compounds, ATC0017 had the lowest MIC value against two types of invasive pathogens and exhibited an antifungal effect. Compounds ATC0018 and ATC0019 showed antifungal effects, but their MIC values ​​were measured to be higher than those of ATC0017, and they did not show a fungicidal effect. ATC0020 showed an antifungal effect against C. auris only against C. albicans, but its MIC value was measured to be higher than that of compounds ATC0017, ATC0018, and ATC0019. ATC0013 did not show an antifungal effect against either of the two types of invasive pathogens. All five additional compounds measured higher MIC values ​​than ATC0001.

[0170] Example 3-4. ATB6015 is based on the optimality of the candidate material 도출을 마이스 장하니다

[0171] To evaluate how much the solubility of five derivatives and salt-form compounds improved compared to ATB6015, the concentration was adjusted to 2.56 mg / ml, the maximum concentration of the drug used in MIC tests. Solubility in DW and PEG400 solvents, commonly used as excipients, was evaluated on a 6-point scale, with 100% of each solvent used (the solubility score was rated on a 6-level scale, ranging from 1 for insolubility to 6 for complete dissolution; 2 for particles remaining and settling or phase separation; 3 for particles remaining but not settling; 4 for fine particles visible; and 5 for cloudy dissolution or a long dissolution time).

[0172] As a result of the solubility evaluation, as shown in the table below, it was confirmed that the solubility of all ATB6015 derivatives and salt-form compounds in DW was improved compared to ATB6015, and the salt-form of ATB6015 showed the most improved solubility with respect to DW with a score of 6. Among the ATB6015 derivatives, ATC0001 free increased the solubility by 2 points compared to ATB6015 in 100% DW solvent when the drug was dissolved at a concentration of 2.56 mg / ml, and completely dissolved in 100% PEG400 solvent, increasing the solubility score by 1 point compared to ATB6015.

[0173] compound DW can be used Do not use PEG400 compound DW Solubility PEG400 Solubility ATB6015 1 5 ATB6015 salt 6 4 ATC0001 free 3 6 ATC0001 salt 2 5 ATC0002 free 2 5 ATC0002 salt 4 2 ATC0003 free 4 2 ATC0003 salt 5 3 ATC0004 free 3 5 ATC0004 salt 2 5 ATC0005 2 2 ATC0005 salt 3 2

[0174] Synthesizing the above experiments, the ATC0001 free MIC value is azole resistance Candida In the strains, values ​​were improved by approximately 10 to 32 times compared to ATB6015, and additionally, MIC antifungal activity was increased by approximately 17 to 66 times in filamentous strains. To identify the target compound with improved solubility compared to ATB6015, solubility was measured; as a result, ATC0001 free showed a 2-point increase in solubility to DW and a 1-point increase in solubility to PEG400 compared to ATB6015. Broad spectrum in vitroBased on the results showing improved antifungal activity and solubility in the most commonly used solvents, ATC0001 free was selected as the optimal candidate.

[0175] Example 4. Confirmation of properties of optimized candidate materials based on ATB6015

[0176] Example 4-1. Evaluation of Physical Properties of Optimization Candidate Material ATC0001 Free

[0177] To evaluate the physical properties (physichochemiocal parameters) of ATC0001 free, solubility, pKa, lipophilicity, and melting temperature were measured by a specialized analytical institution.

[0178] (1) As a result of the solubility evaluation, the intrinsic solubility of ATC0001 free was determined using the pH-metric method (potentiometric method) in the pH range from pH 2 to pH 12, and as shown in the table below, a result of 7.923 μM was obtained.

[0179] compound name Intrinsic Sol. T / ℃ Method Ionic Environment Assay Type ATC0001 free 7.923 μM 25.0 pH-metric 0.168 M KCl CheqSol

[0180] (2) The degree of ionization (pKa) is a property that can predict the physicochemical behavior of a compound and is a parameter that affects lipidity, solubility, protein binding and permeability. As shown in the table below, the pKa of ATC0001 free was found to be 4.94.

[0181] compound name pKa Type T / ℃ Ionic Environment Method ATC0001 free 4.94 ± 0.01 Base 25.0 0.168 M KCl pH-metric

[0182] (3) Lipophilicity (logP / logD) is a physical property that is very related to the absorption of a drug in the human body. It was analyzed by measuring the partition coefficient in water and octanol, and the LogP value of ATC0001 free was measured as 2.86 as follows.

[0183] compound name LogP Type logD at pH 7.4 (blood) T / ℃ Ionic Environment Method ATC0001 free 2.86 ± 0.04 Neutral 2.86 25.0 0.165 M KCl pH-metric

[0184] (4) The melting temperature is an intrinsic property of the compound. When the melting temperature of ATC0001 free was measured using differential scanning calorimetry (DSC), the value was confirmed to be 163℃-165℃ as shown in the table below.

[0185] compound name melting point Analysis method Analysis conditions Sampling / Preprocessing ATC0001 free 167℃ - 169℃ DSC Analysis -10℃ ~ 300℃ range 52 min Standard Sealing Pan(GCA-0052, Aluminum)

[0186] Example 4-2. Comparison of Physical Property Evaluations of ATB6015 and ATC0001 Free

[0187] (1) The intrinsic solubility of ATC0001 free was measured to be 7.923 μM, and the solubility did not increase significantly compared to ATB6015. Further evaluation of solubility showed that ATC0001 free had solubility scores for DW and PEG400 that increased by 2 points and 1 point, respectively, compared to ATB6015. in vivo It was considered that ATC0001 free was more suitable as the compound to be dissolved in the solvent for the experiment.

[0188] compound name Intrinsic Sol. T / ℃ Method Ionic Environment Assay Type ATB6015 94.18 μM 24.5 pH-metric 0.167 M KCl Curve fitting ATB6015 salt 366.6 μM 24.3 pH-metric 0.165 M KCl Curve fitting ATC0001 free 7.923 μM 25.0 pH-metric 0.168 M KCl CheqSol ATC0001 salt 37.89 μM 25.0 pH-metric 0.165 M KCl CheqSol

[0189] (2) In addition, regarding structural characteristics, two pKa values ​​of ATB6015 and one pKa value of ATC0001 free can be analyzed. As a result of analyzing the degree of ionization (pKa) of ATC0001 free, ATB6015 was found to have a value of 4.94 as shown in the table below, which is similar to the pKa values ​​of 3.89 and 5.17 of ATB6015.

[0190] compound name pKa Type T / ℃ Ionic Environment Method ATB6015 3.89 ± 0.05 Base 24.9 0.171 M KCl pH-metric 5.17 ± 0.03 Base 24.9 0.171 M KCl pH-metric ATB6015 salt 3.67 ± 0.01 Base 24.9 0.161 M KCl pH-metric 5.12 ± 0.02 Base 24.9 0.161 M KCl pH-metric ATC0001 free 4.94 ± 0.01 Base 25.0 0.168 M KCl pH-metric ATC0001 salt 4.65 ± 0.04 Base 25.0 0.169 M KCl pH-metric

[0191] (3) Meanwhile, according to Lipinski's rule, a lipophilicity LogP value less than 5 is considered suitable as a drug, and ATB6015's LogP value was 2.41 and ATC0001 free's LogP value was 2.86, so both compounds were determined to be suitable for use as drugs as shown in the table below.

[0192] compound name LogP Type logD at pH 7.4 (blood) T / ℃ Ionic Environment Method ATB6015 2.41 ± 0.03 Neutral 2.40 25.0 0.169 M KCl pH-metric ATB6015 salt 2.07 ± 0.03 Neutral 2.07 25.0 0.171 M KCl pH-metric ATC0001 free 2.86 ± 0.04 Neutral 2.86 25.0 0.165 M KCl pH-metric ATC0001 salt 1.90 ± 0.06 Neutral 1.90 25.0 0.169 M KCl pH-metric

[0193] (4) Melting temperature is an intrinsic property of a compound. When the melting points of ATB6015 and ATC0001 free were measured using differential scanning calorimetry (DSC), values ​​of 163°C-165°C and 167°C-169°C were measured, respectively, and it was confirmed that both compounds had similar levels as shown in the table below.

[0194] compound name melting point Analysis method Analysis conditions Sampling / Preprocessing ATB6015 163℃ - 165℃ DSC Analysis -10℃ ~ 300℃ range, 52 min Standard Sealing Pan (GCA-0052, Aluminum) ATB6015 salt 188℃ - 190℃ DSC Analysis -10℃ ~ 300℃ range, 52 min Standard Sealing Pan (GCA-0052, Aluminum) ATC0001 free 167℃ - 169℃ DSC Analysis -10℃ ~ 300℃ range, 52 min Standard Sealing Pan (GCA-0052, Aluminum) ATC0001 salt 158℃ - 160℃ DSC Analysis -10℃ ~ 300℃ range, 52 min Standard Sealing Pan (GCA-0052, Aluminum)

[0195] Example 4-3. Evaluation of the solubility of optimization candidate substance ATC0001 free

[0196] To identify excipients used in oral and intravenous animal studies, solubility in each solvent was evaluated (the solubility score was rated on a 6-point scale, ranging from 1 for insolubility to 6 for complete dissolution; 2 for particles remaining and settling or phase separation; 3 for particles remaining but not settling; 4 for fine particles visible; and 5 for cloudy dissolution or a long dissolution time).

[0197] ATC0001 was dissolved at a concentration of 100 mg / ml in various excipients such as PEG, Propylene Glycol (PG), Tween, Dimethyl Sulfoxide (DMSO), Carboxymethyl Cellulose (CMC), and Cyclodextrin (CD) to determine the excipient to be used for single-dose toxicity oral administration, and was dissolved at concentrations of 1 to 10 mg / ml to determine the excipient for oral and intravenous administration to be used for efficacy experiments.

[0198] At a level of 10 mg / ml, it dissolved completely under the excipient condition of 10% DMSO in corn oil, and at a concentration of 6 mg / ml, it dissolved best in PEG400 with 50% propylene glycol. At a level of 2 mg / ml, it dissolved in a combination of 5% DMSO, 10% solutol, and 85% PBS, and at a concentration of 1 mg / ml, solubility was high under the condition of 5% DMSO + 10% solutol in DW or PBS, as shown in the table below (Table 22).

[0199] density Brothers Degree of dissolution * 100mg / ml 100% corn oil 3 50% PEG400 in PG 4 10% DMSO in Corn oil 2 50% PEG400 in Corn oil 3 10mg / ml 10% PEG400 in DW 1 30% PEG400 in DW 2 50% PEG400 in DW 3 30% Corn oil in DW 2 50% PG + 2% (Tween 80 1%) in saline 3 50% PG in DW 4 40% CD in water 4 0.5% CMC in water 4 10% DMSO + 40% PEG300 + 5% (Tween 80 1%) in saline 3 10% DMSO in corn oil 6 5% DMSO + 10% PEG400 in CD (40% CD in water) 3 6mg / ml 100% corn oil 5 2% CMC in DW 4 5% DMSO + 2% Tween 80 in DW 3 2% Tween 80 in 0.5% CMC 3 5% DMSO + 2% Tween 80 + 40% PEG in 0.5% CMC 4 5% DMSO + 2% Tween 80 in 0.5% CMC 4 10% DMSO in 0.5% CMC 3 100% olive oil 5 50% Propylene glycol in PEG400 6 4mg / ml 5% DMSO + 10% Solutol in PBS 4 2mg / ml 10% (50% Castor oil + 50% EtOH) in DW 4 5% DMSO + 10% Solutol in PBS 6 1mg / ml 10% (50% Castor oil + 50% EtOH) in DW 5 5% DMSO + 10% Solutol in DW or PBS 5-6

[0200] Example 5. Optimizing candidates Candida spp in vitro active assessment

[0201] Example 5-1. ATC0001 free optimization candidate selection and MFC evaluation

[0202] The Minimum Fungicidal Concentration (MFC) was measured using the ATC0001-free candidate substance to determine the fungicidal (fungicide) or fungistatic (growth inhibition) mode in which the drug kills the bacterial strains. For MFC measurement, wells corresponding to 1X, 2X, 4X, and 8X MICs from the MIC plate were plated onto YPD plates after incubation, and the colonies were counted and compared with the MIC values. The bacterial strains used for MFC measurement were as follows.

[0203] - 2 types of invasive pathogenic fungi ( Candida albicans (SC5314) , Candida tropicalis (KCCM 50075))

[0204] - 2 types of multidrug-resistant strains ( Candida albicans (12-99) , Candida glabrata (BG-2))

[0205] Analysis of MFC revealed that ATC0001 free is an invasive fungus Candida albicans(SC5314) and multidrug-resistant strains Candida albicans (12-99) , Candida glabrata It was determined that it fungistatically inhibits the growth of strains in (BG-2), similar to ATB6015. Invasive pathogens Candida tropicalis In (KCCM 50075), both ATB6015 and ATC0001 free were found to fungicidally kill strains, and in particular, ATC0001 free showed fungicidal antifungal efficacy even at a drug concentration of 4 µg / ml, which is lower than the MFC value of 16 µg / ml for ATB6015.

[0206] Drugs C. albicans (SC5314) C. albicans (12-99) C. glabrata (BG-2) C. tropicalis (KCCM 50075) MIC MFC MIC MFC MIC MFC MIC MFC Amphotericin B 0.06 0.125fungicidal 0.125 0.25fungicidal 0.25 0.25fungicidal 0.13 0.125fungicidal Fluconazole 2 N / A >64 N / A 16 N / A 1 N / A ATB6015 0.25 >2fungistatic 4 >32fungistatic 2 >16fungistatic 4 16fungicidal ATB6015 salt 0.5 >2fungistatic 8 >32fungistatic 2 >16fungistatic 4 8fungicidal ATC0001 free 0.03 >0.25fungistatic 0.13 1fungistatic 0.25 >2fungistatic 1 4fungicidal

[0207] Example 5-2. Measurement of the biofilm inhibitory efficacy of the optimized candidate substance ATC0001-free

[0208] Fungi form a biofilm, a mucous layer composed of a complex mixture of proteins, sugars, and lipids, to protect themselves from drugs or external stimuli; once the biofilm is formed, resistance to drugs increases approximately 100-fold, making it difficult to effectively eliminate the fungi. Therefore, measuring the ability to eliminate fungi that have formed biofilms was crucial during the development of antifungal agents. Multidrug resistance Candida Fungi Candida albicans (12-99) , Candida glabrata The adherence and development inhibition of ATB6015 and ATC0001-free were measured in (BG-2) to compare their biofilm disruption capabilities with those of ATB6015.

[0209] strains resistant to azole drugs Candida albicans Measurement of the biofilm inhibitory activity of ATC0001 free against (12-99) showed that the sessile minimal inhibitory concentration (SMIC) 50% inhibited 50% of the biofilm formed by the strain. 50Compared to ATB6015, the activity inhibiting adherence increased by more than 512 times, and the development inhibitory ability improved by 200 times. The sessile minimal inhibitory concentration (SMIC) inhibiting 90% of biofilms was 90. 90 In ), ATC0001 free increased the biofilm adherence inhibition ability by 200 times compared to ATB6015, and the biofilm development inhibition ability was also improved by 200 times.

[0210] Azole-resistant strains Candida glabrata Analysis of the biofilm inhibitory activity of ATC0001 free against (BG-2) showed that the sessile minimal inhibitory concentration (SMIC) inhibited 50% of the biofilm formed by the strain. 50 Compared to ATB6015, the activity inhibiting adherence increased 200-fold, and the ability to inhibit biofilm development was also improved 200-fold. The sessile minimal inhibitory concentration (SMIC) that inhibits 90% of biofilms is 90. 90 As a result of the measurement, ATC0001 free increased the ability to inhibit biofilm adherence by more than 1,067 times compared to ATB6015, and the degree of inhibition of biofilm development improved by about 267 times.

[0211] Candida albicans Biofilm inhibitory activity of ATC0001 free against (12-99) (Unit: μg / ml) Adherence inhibition SMIC 50 Developmentinhibition SMIC 50 Adherence inhibition SMIC 90 Development inhibition SMIC 90 Amphotericin B 0.125 0.03 1 0.25 Fluconazole >64 64 >64 >64 ATB6015 >64 2 >64 8 ATB6015 salt >64 2 >64 8 ATC0001 free 0.125-0.25 0.01-0.03 0.25 0.03-2 ATC0001 salt 0.125 16 0.03 0.25

[0212] Candida glabrata Biofilm inhibitory activity of ATC0001 free against (BG-2) (Unit: μg / ml) Adherence inhibition SMIC 50 Developmentinhibition SMIC 50 Adherence inhibition SMIC 90 Development inhibition SMIC 90 Amphotericin B 1 0.06 >64 >8 Fluconazole 4 2 >64 16 ATB6015 2 2 >64 8 ATB6015 salt 2 2 64 8 ATC0001 free 0.01 0.01-0.03 0.06-0.125 0.03-0.06 ATC0001 salt 0.25 0.01 4 0.06

[0213] When combining the above information Candida albicans(12-99) In the strain, ATC0001 free improved the concentration that inhibits 50% of the biofilm compared to ATB6015 by about 200 times and 512 times, respectively, and improved the inhibition of 90% of the biofilm by 200 times in both adherence and development methods. Candida glabrata In the (BG2) strain, inhibition of 50% of the biofilm increased activity 200-fold using both adherence and development methods, while inhibition of 90% of the biofilm improved by approximately 1,067-fold and 267-fold, respectively, using adherence and development methods. Therefore, ATC0001-free multidrug resistance compared to ATB6015 Candida The ability to inhibit biofilms produced by fungi was improved by approximately 200 to 1,067 times.

[0214] Example 5-3. Measurement of the emergence of resistant strains of the ATC0001-free candidate substance

[0215] With the recent gradual increase in strains resistant to antifungals, the development of antifungal agents capable of effectively inhibiting the growth of multidrug-resistant fungi is essential. Candida glabrata (ATCC 2001) Performed MIC experiments of compounds against strains to determine the IC values ​​for each drug 50 The values ​​were determined. IC for each drug 50 Resistance was induced by culturing bacteria in media containing concentrations of 1, 4, 8, 16, and 32X. The initial bacterial concentration was 10 6 It was inoculated to this extent, and the number of bacteria increased 100-fold even in the medium containing the drug, 10 8 When it reached [value], it was determined that a resistant strain had appeared; the corresponding date was circled, a sample stock was prepared, and it was stored in a deep freezer. At the same time, 10 in new drug media of different concentrations 6The bacteria were subcultured until the number was sufficient, and this was repeated daily for 40 days. After 40 days, once the measurement of the frequency of resistant strain emergence was completed, the MIC (Amphotercin B and ATB6015 derivatives) and IC of the sample stock were measured. 50 The values ​​of azole drugs (Fluconazole and Efinaconazole) were analyzed and compared to the values ​​before the experiment to determine how much resistance had increased. If resistance had increased, the MIC and IC for the same fungal species were determined. 50 As the value increased, a higher concentration of drug was required to kill the bacteria.

[0216] Results of the test measuring the frequency of appearance of resistant strains, Candida glabrata IC50 when treated with ATB6015-free drug in (ATCC 2001) strains 50 Resistant strains were generated proportionally to concentration, appearing 15 times at 1X, 13 times at 4X, 10 times at 8X, 7 times at 16X, and 5 times at 32X; on average, resistant strains appeared approximately 10 times over 40 days. ATC0001 free IC 50 Similarly, concentration-proportional results were observed, with 12 occurrences at 1X, 4 at 4X, 4 at 8X, 3 at 16X, and 1 at 32X, and an average of about 5 resistant strains appeared over 40 days. ATC0001 free reduced the frequency of appearance of resistant strains by more than twofold compared to ATB6015.

[0217] For sample stocks of resistant strains, the MIC was measured for the three test drugs and the commercially available Amphotericin B drug, and the IC was measured for the azole-class drug. 50 It was measured and shown as a graph in Figure 2.

[0218] The experimental results for the control group were as follows. Sample analysis of the commercial drug Amphotericin B revealed that the MIC values ​​in the 1, 4, and 8X treatment groups did not increase compared to pre-experiment levels, leading to the conclusion that drug tolerance did not increase. Next, regarding the azole drug Fluconazole, the IC values ​​in the control groups were 8-fold at 4X, 4-fold at 8X, and 8-fold at 16X. 50 The values ​​increased, and Efinaconazole showed a 4-fold increase at 4X and a 4-fold increase at 8X IC5. 50 The value increased.

[0219] For the lead compound ATB6015, the MIC values ​​did not increase in the 1X to 32X groups, while for ATB6015 salt, the MIC values ​​doubled in the 4, 8, and 16X groups. For the candidate compound ATC0001 free, the MIC values ​​did not increase compared to the pre-experiment analysis values ​​in all groups from 1X to 32X. As a result of the analysis of resistant strain sample stocks, it was determined that ATC0001 free does not develop resistant strains, as the MIC values ​​did not increase compared to the pre-experiment values, similar to the lead compound ATB6015.

[0220] To summarize the contents of Figure 2 Candida glabrata (ATCC 2001) In the strain, the ATC0001 free candidate substance improved the frequency of appearance of resistant strains by twofold compared to ATB6015, and the results of the MIC analysis of the appearing resistant strains showed that ATC0001 free did not increase the MIC value, similar to ATB6015, and was interpreted as not inducing resistance in the strain.

[0221] Example 6. Optimized candidate substance Candida Regarding spp. in vivo Activity evaluation

[0222] Example 6-1. Azole-resistant strain Candida albicans (12-99) in vivo Efficacy evaluation

[0223] To establish a fungal systemic infection model, male ICR mice Candida albicans (12-99) administered intravenously to the systemic Candida Infection was induced. The antifungal efficacy of the substances was evaluated and compared by administering a commercial drug (Caspofungin) and candidate substances (ATB6015, ATC0001 free) orally or intraperitoneally to an infection model. Survival rates and body weight were measured up to day 15 with 8 animals per group, and the composition of the excipients for the drug and candidate substances was standardized to Castor oil : EtOH : H2O = 5 : 5 : 90 (Table 25, Figs. 3a and 3b).

[0224] army Test Articles Dose Administered amount Route of administration Frequency of administration Administration period Population / Group (mg / kg) (mL / kg) G1 Vehicle - 5 PO QD 5 days 8 G2 Caspofungin 0.1 IP G3 ATB6015 5 PO G4 ATB6015 10 PO G5 ATC0001 free 5 PO G6 ATC0001 free 10 PO G7 ATC0001 free 20 PO

[0225] As a result, as shown in Figures 3a and 3b, the survival rate and body weight changes of mice were observed for 15 days after infection (Day 0). In the control group (G1) without drug administration, the survival rate on Day 15 was 0, with all individuals dying. In contrast, the Day 15 survival rate of the control group administered caspofungin (G2) was 50%, demonstrating a typical antifungal effect comparable to that of a commercially available drug. The experimental results showed equivalent results, with the Day 15 survival rate of the ATB6015 10 mg / kg group (G4) and the ATC0001 free 10 mg / kg group (G6) both at 62.5%; this figure was 12.5% ​​higher than that of caspofungin, which showed a survival rate of 50%. Therefore, ATB6015 and ATC0001 free [drugs] against azole resistance Candida albicans It suggested that it has antifungal efficacy against infected animal models.

[0226] Example 6-2. Azole-resistant strain Candida albicans (12-99) in vivo Efficacy evaluation

[0227] commissioned to another specialized institution, azole-resistant strain Candida albicans (12-99) in vivo We intended to cross-check the efficacy. To establish a systemic fungal infection model, male ICR mice Candida albicansFungal infections were induced by intravenous administration of (12-99). The antifungal efficacy of the substances was evaluated by administering commercial antifungal agents (Caspofungin, Posaconazle) and candidate substances (ATB6015, ATC0001) orally or intraperitoneally to the infection model. Survival rates and body weights were compared with 7 animals per group. The composition of the excipients for the drugs and candidate substances was standardized to Castor oil : EtOH : H2O = 5 : 5 : 90 (Table 27, Figs. 4a and 4b).

[0228] army Test Articles Dose (mg / kg) Administered amount Route of administration Frequency of administration Administration period Population / Group (mL / kg) G1 Vehicle - 5 PO QD 5 days 7 G2 Caspofungin 0.1 IP G3 Posaconazole 2 PO G4 ATB6015 10 PO G5 ATC0001 free 10 PO G6 ATC0001 free 20 PO G7 ATC0001 free 30 PO G8 ATC0001 free 50 PO

[0229] As shown in Figures 4a and 4b, the body weight and condition of mice were continuously tracked for 11 days after infection (Day 0). While the survival rate on day 11 for the control group (G1) without drug administration was 14%, the survival rates on day 11 for the positive control drug caspofungin administration group (G2) and the negative control drug posaconazole administration group (G3) were 100% and 57%, respectively, showing normal antifungal effects of the commercial drugs.

[0230] Furthermore, the survival rate at day 11 for the G5 group (10 mg / kg) administered ATB6015 was 86%, while the survival rates for the ATC0001-free 30 mg / kg group (G7) and the ATC0001-free 50 mg / kg group (G8) were both 100%, representing a 14% increase in survival rate compared to ATB6015. Since the survival rates of the ATC0001-free 30 mg / kg and 50 mg / kg groups were the same as those of the positive control drug caspofungin group (G2), ATB6015 and ATC0001-free azole-class drug resistance Candida albicans It suggested that it has antifungal efficacy against infected animal models.

[0231] Example 6-3. Novel multidrug-resistant fungi Candida auris for in vivo Confirmation of efficacy evaluation

[0232] To establish a fungal systemic infection model, male ICR mice Candida auris Systemic mycosis was induced by intravenous administration. The antifungal efficacy of the substances was evaluated by orally or intraperitoneally administering a commercial antifungal agent (Caspofungin) and candidate substances (ATB6015, ATC0001 free) to the infection model. Survival rates and body weights were measured for 8 animals per group, and the composition of the excipients for the drug and candidate substances was standardized to Castor oil : EtOH : H2O = 5 : 5 : 90 (Table 28, Figs. 5a and 5b).

[0233] army Test Articles Dose (mg / kg) Administered amount Route of administration Frequency of administration Administration period Population / Group (mL / kg) G1 Vehicle - 5 PO QD 5 days 8 G2 Caspofungin 0.1 IP G3 ATB6015 10 PO G4 ATC0001 free 10 PO G5 ATC0001 free 20 PO G6 ATC0001 free 30 PO G7 ATC0001 free 40 PO

[0234] As shown in Figures 5a and 5b, the body weight and condition of mice were tracked and observed for 22 days after infection (Day 0). While the survival rate on day 22 for the control group (G1) without drug administration was 50%, the survival rate on day 22 for the control group administered caspofungin (G2) was 75%, demonstrating the antifungal effect of a standard commercial drug, and the experimental conditions were well established.

[0235] In particular, among the test substance administration groups, the survival rate of the ATC0001 free 20 mg / kg administration group (G5) was 75%, and the survival rate of the ATC0001 free 30 mg / kg administration group (G6) was 62.5%, which was 1.5 times and 1.25 times better than the survival rate of the ATB6015 10 mg / kg administration group (G4), which was 50%.

[0236] Example 7. Evaluation of PK and Stability of Optimized Candidate Substances

[0237] Example 7-1. PK measurement of ATC0001 free

[0238] Pharmacokinetics (PK) were measured to quantitatively determine the processes of absorption, distribution, transport, and excretion over time following administration of the synthetic candidate substance into the body. Using SD rat experimental animals, plasma was collected after administering the candidate substance ATC0001 free orally (per oral, PO) at 10 mg / kg and intravenously (intravenous, IV) at 0.5 mg / kg, and pharmacokinetics (PK) were confirmed.

[0239] compound name Route Dose mg / kg T 1 / 2h T max h C maxng / mL AUC (0-t) h*ng / mL AUC (0- ∞ )h*ng / mL MRT (0-t)h MRT (0- ∞ ) h V z mL / kg Cl mL / h / kg F% ATB6015 IV 2 4.7 0.08 281 1,025 1,049 4.9 5.5 12,880 1,920 N / A PO 10 12.9 4 201 2,416 3,408 8.3 18.2 N / A N / A 47.1 ATB6015 salt PO 10 29.8 4 1,755 23,460 53,604 9.8 41.9 N / A N / A N / A ATC0001free IV 0.5 3.5 0.08 560 1,340 1,510 3.1 4.5 1.72 5.98 N / A PO 10 12.0 4 1,516 17,845 24,363 8.7 17.2 N / A N / A 66.6 ATC0001 salt PO 10 6.2 4 1,285 14,291 15,435 7.1 9.0 N / A N / A N / A

[0240] (T1 / 2: Terminal half-life, Tmax: Time of the maximum, Cmax: Maximum concentration, AUC (0-t): Area under the curve from the time of dosing to the time of the last observation, AUC (0-∞): Area under the curve from the time of dosing to infinity, MRT (0-t): Mean residence time from of dosing to the time of last observation, MRT (0-∞): Mean residence time from of dosing to infinity, Vz: Volume of distribution, CL: Clearance, F: Bioavailability)

[0241] As shown in Table 29 above, when ATC0001 free was administered intravenously to SD rats, the maximum concentration (Cmax) value, which represents the maximum blood concentration, was improved by approximately 2 times compared to ATB6015, even though the administered drug concentration of ATC0001 free was 1 / 4 lower. When administered orally, the maximum blood concentration, Cmax value, was improved by approximately 8 times in ATC0001 free compared to ATB6015.

[0242] As a result of intravenous administration of ATC0001 free, the area under the curve (AUC) value, representing the total blood volume, improved by approximately 1.3 times compared to ATB6015, and showed a value improved by approximately 7.4 times upon oral administration. The mean residence time (MRT) was similar for both compounds when ATC0001 free was administered intravenously and orally compared to ATB6015. Bioavailability (F), an indicator of the amount absorbed into the body after oral administration, showed that ATC0001 free improved by approximately 1.4 times compared to ATB6015, indicating an increase in bioavailability.

[0243] Synthesizing the results above, it was found that most pharmacokinetic indicators, such as the maximum blood concentration (Cmax), total blood volume (AUC), and amount absorbed into the body (F), were significantly improved when the candidate substance ATC0001 free was administered orally and intravenously to experimental animals compared to ATB6015.

[0244] Example 7-2. Stability evaluation results of ATC0001 free

[0245] ATC0001 free was sent to a specialized testing laboratory to evaluate plasma protein binding, plasma stability, metabolic stability, hepatocellular stability, and cardiotoxicity, and the stability of the compound was measured as shown in the table below.

[0246] compound name Species Plasma protein binding% Plasma stability% Metabolismability% hepatocyticstabilityCL int ( μL / min) hERGIC 50 (μM) ATB6015 Human 99.78 N / A - - - Mouse 99.82 N / A - - - ATC0001free Human 98.6 >100 (30min)>100 (120min) 83.3 189.3 (t 1 / 2 ) 3.7 (CL int ) >100 Mouse 97.4 >100 (30min)>100 (120min) 78.2 427 (t 1 / 2 ) 1.6 (CL int )

[0247] The degree of binding of ATC0001 free to human and mouse-derived plasma proteins was evaluated by a specialized institution, and the degree of binding to plasma proteins (bound form %) was measured through LC-MS / MS analysis. The result was calculated by subtracting the free form %) from 100%. The degree of binding of ATC0001 free to plasma proteins was 98.6% in humans and 97.4% in mice, while ATB6015 showed similar values ​​for both compounds, with 99.78% in humans and 99.82% in mice.

[0248] To evaluate stability in plasma, human and mouse plasmas were treated with the drug and cultured, and the drug was analyzed via LC-MS / MS. Unlike ATB6015, which could not be measured by LC-MS / MS due to ion suppression, ATC0001 free was measurable and maintained similar values ​​of >100% at 30 minutes and 2 hours. Since a plasma stability value of approximately 85% or higher is considered stable, it was interpreted that ATC0001 free has high in vivo stability.

[0249] In addition, the metabolic stability of the drug was measured using liver microsomes to determine the extent to which the compound is metabolized in the liver. ATC0001 free showed metabolic stability values ​​of 83.3% in humans and 78.2% in mice. Since a metabolic stability value of 70–90% is evaluated as a stable compound with a half-life of about 1–3 hours, ATC0001 free was determined to be a stable drug based on the results of the metabolic stability test.

[0250] Additionally, the hepatocyte stability of ATC0001-free was analyzed, and a half-life of 189.3 hours was obtained in human hepatocytes, while a half-life of 427 hours was measured in mouse hepatocytes. Clearance criteria (Unit: μL / min / 10 6 In human hepatocytes, a clearance value of 3.5 or less indicates slow metabolism, while a value of 19 or more indicates rapid metabolism; thus, ATC0001 free, which showed a value of 3.7, could be interpreted as indicating moderate metabolism in humans. In mouse hepatocytes, a clearance value of 3.3 or less indicates slow metabolism, and a value of 17.8 or more indicates rapid metabolism; thus, ATC0001 free showed a value of 1.6, which could be evaluated as having a low metabolic rate in mice.

[0251] Next, we evaluated the hERG cardiotoxicity test, which assesses the potential for arrhythmia induction by measuring the degree of inhibition of hERG potassium channel activity. In the hERG test, hERG-expressing HEK cells were used without separating human and mouse species, and the measurement results showed the IC50 of the ATC0001-free drug. 50 It was evaluated as stable because the value was >100, indicating almost no inhibition of hERG.

[0252] When summarizing the results of the stability evaluation of the above ATC0001 free, it was determined to be stable as a result of measuring plasma stability and metabolic stability by commissioning a specialized testing institution, and as a result of the hepatocyte stability analysis, it showed a moderate metabolic rate in human hepatocytes and did not inhibit hERG under test conditions (100 μM), so it was determined to be a stable compound that does not cause cardiotoxicity.

[0253] Foregoing, specific parts of the present invention have been described in detail. It is evident to those skilled in the art that such specific descriptions are merely preferred embodiments and do not limit the scope of the invention. Accordingly, the actual scope of the invention is defined by the appended claims and their equivalents. Industrial applicability

[0254] Since the compound of the present invention has a high antifungal effect with low side effects, an antifungal pharmaceutical composition containing the same is provided.

Claims

Claim 1 Compounds represented by the following Chemical Formula 1, isomers thereof, or pharmaceutically acceptable salts: [Chemical Formula 1] In the above formula, W is O or S; R1 is hydrogen or a C1-6 alkyl; R2 is -OX-R3; X is a C1-6 alkyl; and R3 is a cycloalkyl group having 5 to 8 carbon atoms, an aryl group, or a monocyclic or dicyclic heteroaryl group comprising one or two heteroatoms independently selected from nitrogen, oxygen, and sulfur, wherein in each case these groups are unsubstituted or one or two halogens, CF 3, It can be substituted by an ethoxy group; Y is substituted by one or more amino groups, and is a heteroaryl or C1 to C comprising one or two heteroatoms independently selected from nitrogen, oxygen, and sulfur. 10 It is an alkylamino group. Claim 2 In claim 1, W is O; R1 is hydrogen; X is a C1-3 lower alkyl; and R3 is a cycloalkyl group having 5 to 8 carbon atoms, a phenyl group, or a monocyclic heteroaryl group containing nitrogen, and in each case, these groups are unsubstituted or have one or two halogens, CF 3, It can be substituted by a methoxy group; Y is substituted by one amino group, and is a heteroaryl or C1 to C comprising one or two nitrogen heteroatoms. 10 A compound, its isomer, or a pharmaceutically acceptable salt that is an alkylamino group of Claim 3 In paragraph 1, W is O; R1 is hydrogen; X is a methyl group; and R3 is a cycloalkyl group having 5 to 8 carbon atoms, a phenyl group, or a pyridyl group, and in each case, these groups are unsubstituted or have one or two halogens, CF 3, A compound, isomer thereof, or pharmaceutically acceptable salt thereof, which can be substituted by a methoxy group; Y being substituted by one amino group and comprising one nitrogen heteroatom, a heteroaryl or C1 to C6 alkylamino group. Claim 4 In claim 1, the compound represented by Formula 1 is a compound selected from the group consisting of compounds represented by Formulas 2 to 20 below, isomers thereof, or pharmaceutically acceptable salts: [Formula 2] [Chemical Formula 3] [Chemical Formula 4] [Chemical Formula 5] [Chemical Formula 6] [Chemical Formula 7] [Chemical Formula 8] [Chemical Formula 9] [Chemical Formula 10] [Chemical Formula 11] [Chemical Formula 12] [Chemical Formula 13] [Chemical Formula 14] [Chemical Formula 15] [Chemical Formula 16] [Chemical Formula 17] [Chemical Formula 18] [Chemical Formula 19] [Chemical Formula 20] Claim 5 A pharmaceutical composition for the prevention or treatment of fungal infections comprising, as an active ingredient, a compound according to any one of claims 1 to 4, an isomer thereof, or a pharmaceutically acceptable salt thereof. Claim 6 A pharmaceutical composition according to claim 5, having antibacterial activity against bacteria selected from Candida, Sportricum, Malassezia, Cryptococcus, Aspergillus, Pneumocystis, and Mucor. Claim 7 A pharmaceutical composition according to claim 6, wherein the fungal infection is selected from meningitis, encephalitis caused by meningitis, cutaneous candidiasis, mucosal candidiasis, visceral candidiasis, urinary tract candidiasis, candidal endocarditis, oropharyngeal candidiasis, candidal endophthalmitis, candidal sepsis, ringworm, tinea versicolor, sportrixosis, fungal abscess, fungal granuloma, pyogenic granuloma, Madura mycosis, Pneumocystis pneumonia, systemic cryptococcosis, cutaneous and mucosal cryptococcosis, aspergillosis, cavities, hemoptysis, allergy, chronic pulmonary tuberculosis, pulmonary fibrosis, pulmonary cysts, chronic fever, cough, sputum, hemoptysis, ringworm, thrush, fungal dementia, and zygomycosis. Claim 8 A method for preventing or treating mycosis comprising administering to a subject a compound according to any one of claims 1 to 4, an isomer thereof, or a pharmaceutically acceptable salt thereof. Claim 9 A method according to claim 8, having antibacterial activity against bacteria selected from Candida, Sportricum, Malassezia, Cryptococcus, Aspergillus, Pneumocystis, and Mucor. Claim 10 In claim 9, the above fungal infection is selected from meningitis, encephalitis caused by meningitis, cutaneous candidiasis, mucosal candidiasis, visceral candidiasis, urinary tract candidiasis, candidal endocarditis, oropharyngeal candidiasis, candidal endophthalmitis, candidal sepsis, ringworm, tinea versicolor, sportrixosis, fungal abscess, fungal granuloma, pyogenic granuloma, Madura mycosis, Pneumocystis pneumonia, systemic cryptococcosis, cutaneous and mucosal cryptococcosis, aspergillosis, cavities, hemoptysis, allergy, chronic pulmonary tuberculosis, pulmonary fibrosis, pulmonary cysts, chronic fever, cough, sputum, hemoptysis, ringworm, thrush, fungal dementia, and zygomycosis.

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