Derivative compound comprising amino nicotinamide structure and antifungal pharmaceutical composition comprising same

Abstract

The present invention relates to a derivative compound comprising an amino nicotinamide structure, an isomer thereof or a pharmaceutically acceptable salt thereof, and an antifungal pharmaceutical composition comprising same. More specifically, the present invention provides a benzyloxybenzylamine amino acid derivative having few side effects, and an antifungal agent comprising same. The antifungal agent has a range having antifungal activity against fungi selected from Candida sp., Sporothrix sp., Malassezia sp., Cryptococcus sp., Aspergillus sp., Pneumocystis sp., and Mucor sp.

Description

Derivative compound comprising aminonicotinamide structure and antifungal pharmaceutical composition comprising same

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

[0002] This invention was supported by the Ministry of Health and Welfare under the Infectious Disease Prevention and Treatment Technology Development Project, under the title of "Optimization and Validation of Antifungal Drug-Resistant Candida Infection Treatment Preclinical Trial", with Amtix Bio Co., Ltd. as the lead organization and the Korea Health Industry Development Institute as the research management organization, from August 1, 2022 to December 31, 2023.

[0003]

[0004] Fungal infections occur very frequently in patients whose immune function is weakened due to excessive use of broad-spectrum antibacterial agents, organ transplantation, long-term administration of anticancer drugs, aging, AIDS, or procedures such as catheter or prosthetic devices.

[0005] Conventional azole drugs can cause death due to 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, and may cause enterocolitis. These side effects are exacerbated in patients with impaired renal function, so renal function monitoring is crucial. It is also contraindicated in pregnant women. The main toxicity of amphotericin B is glomerular nephrotoxicity due to renal artery constriction. This is dose-dependent, and lifetime cumulative doses exceeding 4-5 g increase the risk of permanent renal function loss. Furthermore, nephrotoxicity, including excessive loss of potassium, magnesium, and bicarbonate due to renal tubular toxicity and decreased production of hematopoietic hormones, can occur. In addition, acute reactions such as thrombophlebitis, chills, tremors, and hyperventilation may occur. In this way, existing antifungal agents show various side effects depending on the type of drug, and there is a need to develop new treatments that can increase antifungal effects while reducing these side effects.

[0006]

[0007] The present invention aims to provide a derivative compound comprising an aminonicodinamide structure, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof; and an antifungal composition comprising the same as an active ingredient, which inhibits the formation of a fungal biofilm and / or the formation of antifungal-resistant bacteria.

[0008]

[0009] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by those skilled in the art from the description below.

[0010]

[0011] Various embodiments of the present invention are described with reference to the drawings. In the following description, various specific details, such as specific configurations, compositions, and processes, are set forth to provide a thorough understanding of the present invention. However, certain embodiments may be practiced without one or more of these specific details, or in conjunction with other known methods and configurations. In other instances, well-known processes and manufacturing techniques are not described in specific detail so as not to unnecessarily obscure the present invention. Reference throughout this specification to an embodiment means that a particular feature, configuration, composition, or characteristic described in connection with the embodiment is included in one or more embodiments of the present invention. Therefore, the appearance of an embodiment in various places throughout this specification does not necessarily indicate the same embodiment of the present invention. Additionally, the particular features, configurations, compositions, or characteristics may be combined in any suitable manner in one or more embodiments. In the present invention, unless otherwise specified, terms used have their ordinary meanings well 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 racemate, or cis-trans isomers of alkenes, etc.).

[0012] In one specific embodiment, a compound represented by the following formula (I), an isomer thereof or a pharmaceutically acceptable salt thereof is provided:

[0013] [Chemical Formula I]

[0014]

[0015] In the above formula, W is O or S; R1 is hydrogen or C1-6 alkyl; R2 is -OX-R3; X is C1-6 alkyl; R3 is a cycloalkyl group having 5 to 8 carbon atoms, an aryl group or a monocyclic or bicyclic heteroaryl group containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen and sulfur, which in each case may be unsubstituted or substituted by 1 or 2 halogens or CF3; Y is NH2, OH, OMe, or OEt. In the above specific examples, W is O; R1 is hydrogen; X is C1-3 lower alkyl; R3 is a cycloalkyl group having 5 to 8 carbon atoms, a phenyl group or a monocyclic heteroaryl group containing nitrogen, which in each case may be unsubstituted or substituted by 1 or 2 halogens or CF3; Y is NH2, providing a compound, an isomer thereof or a pharmaceutically acceptable salt thereof, wherein in said specific embodiment, W is O; R1 is hydrogen; X is a methyl group; 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.

[0016] In one specific example, a pharmaceutical composition for preventing or treating mycosis is provided, comprising the compound, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient. In the above specific example, a pharmaceutical composition having antibacterial activity against a fungus selected from the group consisting of Candida, Sportricum, Malassezia, Cryptococcus, Aspergillus, Pneumocystis, and Mucor is provided, and in the above specific example, the mycosis is meningitis, encephalitis due to meningitis, cutaneous candidiasis, mucosal candidiasis, visceral candidiasis, urinary candidiasis, candidal endocarditis, oropharyngeal candidiasis, candidal endophthalmitis, candidal sepsis, tinea, tinea versicolor, sporriosis, fungal abscess, fungal granuloma, pyogenic granuloma, madura mycosis, Pneumocystis carinii pneumonia, systemic cryptococcosis, mucocutaneous cryptococcosis, aspergillosis, cavities, hemoptysis, allergies, and encephalitis. A pharmaceutical composition is provided, wherein the composition is selected from among pulmonary tuberculosis, pulmonary fibrosis, pulmonary cysts, chronic fever, cough, sputum, bloody sputum, jock itch, thrush, fungal dementia and zygomycosis.

[0017] In the present invention, the halogen is F, Cl, Br or I.

[0018] In the present invention, the term “C 1-7 “Alkyl” means a straight or branched chain alkyl having 1 to 7 carbon atoms, including but not limited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl; preferably ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, and tert-butyl.

[0019] In the present invention, the term “cycloalkyl” means a cyclic alkyl having three or more carbon atoms in the ring, and includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and cyclodecyl. The terms “C3-C8cycloalkyl,” “C3-C7cycloalkyl,” and “C3-C6cycloalkyl” have similar meanings.

[0020] In the present invention, the term “aromatic heterocycle” or “heteroaryl” has the same meaning and refers to a heteroaromatic group containing one or more heteroatoms. The heteroatoms referred to herein include oxygen, sulfur and nitrogen. Examples thereof include furyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl and the like. The heteroaryl ring may be fused to an aryl, a heterocyclic group or a cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring. The heteroaryl is optionally substituted or unsubstituted.

[0021] In the present invention, the term "substitution" means that one or more hydrogen atoms on a specified group are replaced by a specified substituent. The specified substituent is a substituent described correspondingly above or a substituent shown in each embodiment. Unless specifically stated otherwise, a specified substituted group may have one substituent selected from a specified group at any substitutable site of the group, and the substituents may be the same or different at each position. A cyclic substituent such as a heterocycloalkyl may be connected to another ring such as a cycloalkyl to form a spiro bicyclic system such as two rings having one common carbon atom. It should be understood by those skilled in the art that the combinations of substituents envisaged in the present invention are stable or chemically feasible combinations. The substituents may be, for example, C 1-8 Alkyl, C 2-8 Alkenyl, C 2-8 alkynyl, 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.

[0022] In one embodiment of the present invention, a compound represented by the following chemical formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof is provided:

[0023] [Chemical Formula 1]

[0024]

[0025] In the above formula, W is O or S; R1 is hydrogen or C1-6 alkyl; R2 is -OX-R3; X is C1-6 alkyl; R3 is a cycloalkyl group having 5 to 8 carbon atoms, an aryl group or a monocyclic or bicyclic heteroaryl group containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen and sulfur, in each case, these groups are unsubstituted or substituted with 1 or 2 halogens, CF 3, may be substituted by an ethoxy group; Y is substituted by one or more amino groups and is heteroaryl or C1 to C containing one or two heteroatoms independently selected from nitrogen, oxygen and sulfur. 10 is an alkylamino group.

[0026] The term "alkylamino" as used herein refers to a group having the structure -NHR', wherein R' is hydrogen, alkyl or substituted alkyl, cycloalkyl or substituted cycloalkyl as defined above.

[0027] The term "dialkylamino" as used herein refers to a group having the structure -NRR', wherein R and R' are each independently alkyl or substituted alkyl, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted cycloalkenyl, aryl or substituted aryl, heterocyclyl or substituted heterocyclyl as defined above. R and R' may be the same or different dialkyamino moieties. 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, and the like. In certain embodiments, R and R' are linked 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.

[0028] In another embodiment of the present invention, W is O; R1 is hydrogen; X is C1-3 lower alkyl; R3 is a cycloalkyl group having 5 to 8 carbon atoms, a phenyl group or a monocyclic heteroaryl group containing nitrogen, in each case these groups are unsubstituted or substituted with one or two halogens, CF 3, may be substituted by a methoxy group; Y is substituted by one amino group and is heteroaryl or C1 to C containing one or two nitrogen heteroatoms 10 Provided is a compound, an isomer or a pharmaceutically acceptable salt thereof, which is an alkylamino group.

[0029] In another embodiment of the present invention, W is O; R1 is hydrogen; X is a methyl group; R3 is a cycloalkyl group, a phenyl group or a pyridyl group having 5 to 8 carbon atoms, in each case these groups are unsubstituted or substituted with one or two halogens, CF 3, A compound, an isomer or a pharmaceutically acceptable salt thereof, wherein Y is substituted with one amino group and is a heteroaryl or C1 to C6 alkylamino group containing one nitrogen heteroatom.

[0030] The pharmaceutical composition of the present invention comprises a compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier within a safe and effective amount range. Here, the “safe and effective amount” refers to an amount of the compound that can significantly improve a disease without causing serious side effects. Typically, the pharmaceutical composition contains 1 to 2,000 mg of the compound / agent of the present invention, more preferably 5 to 500 mg of the compound / agent of the present invention. Preferably, the “first agent” is a capsule or a pill.

[0031] "Pharmaceutically acceptable carrier" 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. "Compatibility" herein means that each component of the composition can be mixed with the compound of the present invention without significantly reducing the efficacy of the compound. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (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, flavors, stabilizers, antioxidants, preservatives, and pyrogen-free water.

[0032] As used herein, the term "optical isomer" or "stereoisomer" refers to any of the various stereoisomeric configurations that may exist for a given compound of the present invention, including geometric isomers. It is understood that substituents may be attached to chiral centers of carbon atoms. The term "chiral" refers to a molecule that has the property of non-superimposability on its mirror image partner, whereas the term "achiral" refers to a molecule that is superimposable on its mirror image partner. Accordingly, the present invention includes enantiomers, diastereomers, or racemates of a compound. "Enantiomers" are a pair of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. The term is used to designate a racemic mixture, 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. When a compound is a pure enantiomer, the stereochemistry at each chiral carbon can be specified by R or S. Resolved compounds whose absolute configuration is unknown can be designated (+) or (-) according to the direction (dextrorotatory or levorotatory) in which they rotate plane-polarized light at the wavelength of the sodium D line. Certain compounds described herein contain one or more asymmetric centers or axes and thus can produce enantiomers, diastereomers, and other stereoisomeric forms that can be defined as (R)- or (S)- in terms of absolute stereochemistry.

[0033] 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 topical administration.

[0034] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or compatibilizers, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; (b) adhesives, such as hydroxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and gum acacia; (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 enhancers, such as quaternary amine compounds; (g) humectants, such as cetanol and glyceryl monostearate; (h) adsorbents such as kaolin; and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate or mixtures thereof. In capsules, tablets and pills, the formulation may also include buffering agents.

[0035] Solid dosage forms, such as tablets, dragees, capsules, pills, and granules, can be prepared using coating and shell materials, such as enteric coatings and other materials known in the art. These may include opacifying agents, and the active compound or compounds in these compositions can be released in a delayed manner at any location within the digestive tract. Examples of usable embedding components include polymeric materials and wax-based materials. If desired, the active compound may be formed into microcapsules with one or more of the above excipients.

[0036] Liquid formulations for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid formulations may contain inert diluents 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.

[0037] In addition to these inert diluents, the composition may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents and fragrances.

[0038] In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan, microcrystalline cellulose, aluminum methoxide and agar or mixtures of these substances.

[0039] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous aqueous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0040] 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 preservatives, buffers, or propellants that may be required.

[0041] 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 another small molecule compound or by forming an ADC with another large molecule compound, such as a monoclonal antibody.

[0042] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is administered to a mammal (e.g., a human) in need of treatment, wherein the dosage upon administration is a pharmaceutically recognized effective dosage. For a human weighing 60 kg, the daily dosage is typically 1 to 2,000 mg, preferably 5 to 500 mg. Of course, the specific dosage should take into account factors such as the route of administration and the patient's health condition, all of which are within the scope of the capabilities of a skilled physician.

[0043] In one specific example, fungi are a group of eukaryote organisms with cell walls, including but not limited to yeasts, molds, and mushrooms. Fungal cell walls contain unique components such as chitin and beta-glucan, making them targets for the development of antifungal agents targeting fungi. Furthermore, a key target is ergosterol, a component of the fungal cell membrane. For example, the Azoles family disrupts the cell membrane by inhibiting ergosterol synthesis. Furthermore, drugs such as flucytosine inhibit fungal cell proliferation by interfering with DNA and RNA synthesis, while some antifungals target ribosomes to block fungal protein synthesis, and drugs such as griseofulvin inhibit cell division by interfering with microtubule function.

[0044] In the present invention, “prevention” may include, without limitation, any act of blocking, suppressing, or delaying disease symptoms by using the pharmaceutical composition of the present invention.

[0045] In the present invention, “treatment” and “improvement” may include without limitation any action that improves or benefits disease symptoms by examining the pharmaceutical composition of the present invention.

[0046] The "pharmaceutical composition" of the present invention is not limited to these, but may be formulated and used in the form of oral dosage forms such as powders, granules, capsules, tablets, aqueous suspensions, etc., external preparations, suppositories, and sterile injectable solutions, each according to a conventional method. The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, coloring agents, fragrances, etc. for oral administration, and buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers, etc. for injections. In the case of topical administration, bases, excipients, lubricants, preservatives, etc. may be used. The formulation 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 injections, it can be manufactured in the form of unit dose ampoules or multiple doses. In addition, it can be formulated as a solution, suspension, tablet, capsule, sustained-release preparation, etc. Meanwhile, examples of carriers, excipients, and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, or mineral oil. Additionally, fillers, anti-coagulants, lubricants, humectants, fragrances, emulsifiers, preservatives, etc. may be included.

[0047] In the present invention, the "administration route" for the pharmaceutical composition includes, but is not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, or rectal administration. Oral or parenteral administration is preferred.

[0048] In the present invention, "parenteral" includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, 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.

[0049] The "dosage and administration" of the composition of the present invention may vary depending on various factors including the activity of the specific compound used, age, body weight, general health, sex, diet, administration time, administration route, excretion rate, drug combination, and severity of the specific disease to be prevented or treated, and the dosage of the pharmaceutical composition may vary depending on the patient's condition, body weight, degree of disease, pharmaceutical form, administration route, and period, but may be appropriately selected by those skilled in the art, and may be administered at 0.0001 to 50 mg / kg or 0.001 to 50 mg / kg per day. The administration may be administered once a day or divided into several times. The above 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, a sugar-coated tablet, a capsule, a liquid, a gel, a syrup, a slurry, or a suspension.

[0050] In one embodiment of the present invention, a compound represented by the following chemical formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof is provided:

[0051] [Chemical Formula 1]

[0052]

[0053] In the above formula, W is O or S; R1 is hydrogen or C1-6 alkyl; R2 is -OX-R3; X is C1-6 alkyl; R3 is a cycloalkyl group having 5 to 8 carbon atoms, an aryl group or a monocyclic or bicyclic heteroaryl group containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen and sulfur, in each case, these groups are unsubstituted or substituted with 1 or 2 halogens, CF 3, may be substituted by an ethoxy group; Y is substituted by one or more amino groups and is heteroaryl or C1 to C containing one or two heteroatoms independently selected from nitrogen, oxygen and sulfur. 10 is an alkylamino group.

[0054] In another embodiment of the present invention, W is O; R1 is hydrogen; X is C1-3 lower alkyl; R3 is a cycloalkyl group having 5 to 8 carbon atoms, a phenyl group or a monocyclic heteroaryl group containing nitrogen, in each case these groups are unsubstituted or substituted with one or two halogens, CF 3, may be substituted by a methoxy group; Y is substituted by one amino group and is heteroaryl or C1 to C containing one or two nitrogen heteroatoms 10 Provided is a compound, an isomer or a pharmaceutically acceptable salt thereof, which is an alkylamino group.

[0055] In another embodiment of the present invention, W is O; R1 is hydrogen; X is a methyl group; R3 is a cycloalkyl group, a phenyl group or a pyridyl group having 5 to 8 carbon atoms, in each case these groups are unsubstituted or substituted with one or two halogens, CF 3, A compound, an isomer or a pharmaceutically acceptable salt thereof, wherein Y is substituted with one amino group and is a heteroaryl or C1 to C6 alkylamino group containing one nitrogen heteroatom.

[0056] In another embodiment of the present invention, the compound represented by the above formula 1 provides a compound selected from the group consisting of compounds represented by the following formulae 2 to 20, an isomer thereof, or a pharmaceutically acceptable salt thereof:

[0057] [Chemical Formula 2]

[0058]

[0059] [Chemical Formula 3]

[0060]

[0061] [Chemical Formula 4]

[0062]

[0063] [Chemical Formula 5]

[0064]

[0065] [Chemical Formula 6]

[0066]

[0067] [Chemical Formula 7]

[0068]

[0069] [Chemical Formula 8]

[0070]

[0071] [Chemical Formula 9]

[0072]

[0073] [Chemical Formula 10]

[0074]

[0075] [Chemical Formula 11]

[0076]

[0077] [Chemical Formula 12]

[0078]

[0079] [Chemical Formula 13]

[0080]

[0081] [Chemical Formula 14]

[0082]

[0083] [Chemical Formula 15]

[0084]

[0085] [Chemical Formula 16]

[0086]

[0087] [Chemical Formula 17]

[0088]

[0089] [Chemical Formula 18]

[0090]

[0091] [Chemical Formula 19]

[0092]

[0093] [Chemical Formula 20]

[0094]

[0095] In another embodiment of the present invention, a pharmaceutical composition for preventing or treating mycosis is provided, comprising the compound, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

[0096] In another embodiment of the present invention, a pharmaceutical composition is provided, wherein the pharmaceutical composition has antibacterial activity against bacteria selected from the group consisting of Candida spp., Sporetricum spp., Malassezia spp., Cryptococcus spp., Aspergillus spp., Pneumocystis spp., and Mucor spp.

[0097] In another embodiment of the present invention, a pharmaceutical composition is provided, wherein the mycosis is selected from meningitis, encephalitis due to meningitis, cutaneous candidiasis, mucosal candidiasis, visceral candidiasis, urinary candidiasis, candidal endocarditis, oropharyngeal candidiasis, candidal endophthalmitis, candidal sepsis, tinea versicolor, sporriosis, fungal abscess, fungal granuloma, pyogenic granuloma, madura mycosis, Pneumocystis carinii pneumonia, systemic cryptococcosis, mucocutaneous cryptococcosis, aspergillosis, cavities, hemoptysis, allergies, chronic pulmonary tuberculosis, pulmonary fibrosis, pulmonary cysts, chronic fever, cough, sputum, bloody sputum, jock itch, thrush, fungal dementia and zygomycosis.

[0098]

[0099] The compound of the present invention has high antifungal effect with low side effects.

[0100]

[0101] Figure 1 compares and analyzes the in vivo efficacy of fluconazole (FCZ) and ATB6015.

[0102] Figure 2 shows the measurement of the frequency of appearance of resistant strains in Candida glabrata.

[0103] Figure 3 shows the change in body weight and survival rate after ATC0001 free administration in the Candida albicans (12-99) infection model.

[0104] Figure 4 is the result of cross-checking by request to another specialized institution, and is about the change in body weight and survival rate after ATC0001 free administration in the Candida albicans (12-99) infection model.

[0105] Figure 5 shows the body weight change and survival rate after drug treatment in the Candida auris infection model.

[0106] Figure 6 shows the results of in vitro activity screening of five additionally synthesized compounds (ATC0013, ATC0017, ATC0018, ATC0019, ATC0020).

[0107] Figure 7a is the spectroscopic data of ATB6015.

[0108] Figure 7b is the spectroscopic data of ATC0001.

[0109] Figure 7c is the spectroscopic data of ATC0002.

[0110] Figure 7d is the spectroscopic data of ATC0003.

[0111] Figure 7e is the spectroscopic data of ATC0004.

[0112] Figure 7f is the spectroscopic data of ATC0005.

[0113] Figure 7g is the spectroscopic data of ATC0006.

[0114] Figure 7h is the spectroscopic data of ATC0007.

[0115] Figure 7i is the spectroscopic data of ATB6015 chloride.

[0116] Figure 7j is the spectroscopic data of ATC0001 chloride.

[0117] Figure 7k is the spectroscopic data of ATC0002 chloride.

[0118] Figure 7l is the spectroscopic data of ATC0003 chloride.

[0119] Figure 7m is the spectroscopic data of ATC0004 chloride.

[0120] Figure 7n is the spectroscopic data of ATC0005 chloride.

[0121] Figure 7o is the spectroscopic data of ATC0013.

[0122] Figure 7p is the spectroscopic data of ATC0017.

[0123] Figure 7q is the spectroscopic data of ATC0018.

[0124] Figure 7r is the spectroscopic data of ATC0019.

[0125] Figure 7s is the spectroscopic data of ATC0020.

[0126]

[0127] Hereinafter, the present invention will be described in more detail through examples. These examples are intended solely to illustrate the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples, in accordance with the gist of the present invention.

[0128]

[0129] Example 1. In vivo drug efficacy evaluation and optimization of ATB6015 using a C. auris infection animal model.

[0130] Example 1-1. In vitro antifungal efficacy of ATB6015, a derivative based on ATB1651, against three Candida species.

[0131] The present inventors prepared a new derivative of a conventional amino acid derivative (ATB1651) having excellent pharmacological properties and antifungal activity based on the structure-activity relationship (SAR) of the following chemical formula 2 (ATB6015):

[0132] [Chemical Formula 2]

[0133]

[0134] The above derivative was confirmed to exhibit antifungal activity that was 4 to 25 times superior in vitro against three Candida species (C. albicans wild type strain SC5314, azole antifungal resistant C. glabrata BG2 strain, and multidrug resistant C. auris B8441 strain) including antifungal drug resistance, as shown in Table 1 below.

[0135] ATB6015, a derivative based on ATB1651 Candida Antibacterial activity against 3 strainsDrugMIC (㎍ / mL)C. albicans(SC5314)C. glabrata(BG-2)C. auris(B8441)Amphotericin B0.250.50.5Fluconazole1>64>64ATB1651488ATB60150.2520.5

[0136]

[0137] Example 1-2. Establishment of a mouse model infected with multidrug-resistant Candida auris and confirmation of the in vivo antifungal efficacy of ATB6015.

[0138] To establish an animal model of C. auris infection to evaluate the systemic infection drug efficacy of ATB6015, 7-week-old female A / J mice were each inoculated with 1x10 C. auris cells. 7 , 5x10 7 , 1x10 8 , 2x10 8 After infection by intravenous administration, the survival was confirmed. The number of infected cells was 1x10 7 Except in the case of , all mice died at 2 days after infection, and the number of infected cells was 1x10 7 was selected.

[0139] Next, to confirm the therapeutic efficacy of ATB6015 on mice infected with Candida auris, the previously set cell concentration (1x10 7 ) were infected with C. auris and the in vivo efficacy of fluconazole (FCZ), a known antifungal agent, and ATB6015 was compared and analyzed (see Fig. 1). After infection, ATB6015 was administered orally five times at a concentration of 10 mpk (3, 24, 48, 60, and 72 hours after infection). As a result, the median survival days of the vehicle-untreated group and the fluconazole-treated group were 6 and 11.5 days, respectively, but the median survival days of the ATB6015-treated group were confirmed to increase significantly to 20 days.

[0140]

[0141] Example 1-3. Drug target discovery of ATB6015 using the HIP assay (Haploinsufficiency Profiling assay)

[0142] After treatment with ATB6015, HIP assays revealed a group of drug target candidates with significantly reduced competitive fitness and low p-values. Among these, GWT1, which had a log2 fold change of -2 or less and the lowest p-value, is known to be involved in glycosyl-phosphatidylinositol (GPI) synthesis, which is related to cell wall integrity (Watanabe et al., 2012, Antimicrob Agents Chemother.), and was therefore judged to be most likely a drug target of ATB6015.

[0143]

[0144] Example 2. Evaluation of physical properties of ATB6015

[0145] Example 2-1. Solubility Evaluation of ATB6015

[0146] For toxicity tests for oral (PO) and intravenous injection (IV) administration, and for future drug development as an injectable drug, drug solubility is important. Therefore, to evaluate the solubility of ATB6015, an antifungal candidate, ATB6015 was dissolved in various excipients used as excipients in animal testing and the solubility in each solvent was evaluated. The solubility score was evaluated on a 6-level scale, with 1 being insoluble and 6 being completely dissolved. The level at which particles remain and settle or layer separation occurs was evaluated as 2, if some particles remain but do not settle, 3, if fine particles are visible, 4, and if it is completely dissolved or takes a long time to dissolve, it was evaluated as 5.

[0147] When ATB6015 was dissolved at a level of 6 mg / ml in various excipients such as PEG, Tween, and Cyclodextrin (CD) for intravenous injection toxicity testing, as shown in Tables 2 and 3 below, ATB6015 was insoluble in water and particles remained in other excipients, settling, or layer separation occurred (Table 2). In addition, even when dissolved at a high concentration of 100 mg / ml for a single oral administration toxicity test, it was hardly dissolved, so it was determined that it was desirable to increase the activity or solubility of the drug (Table 3).

[0148] Solubility evaluation of ATB6015 in various excipientsExcipient solubility * 0.9% saline21 95% DW2ATB6015 : CD = 1:1(w / w) mixed + D.W2(ATB6015 : CD = 1:1(w / w)) : (10% PEG300 + 90% DW) = 1:125% PEG400 + 95% DW25% PEG400 + 95% Saline210% PEG400 + 90% Saline2100mg / kg Glycerol in DW340% CD in DW4

[0149] Excipient solubility * 10% (50% Castor oil + 50% EtOH) + 90% DW150% (50% Castor oil + 50% EtOH) + 50% DW150% PEG400 + 50% DW150% propylene glycol + 50% DW11% Tween80 + 99% DW35% ethanol + 95% DW11% DMSO + 99% DW2

[0150]

[0151] Example 2-2. Measurement of physicochemical parameters of ATB6015

[0152] Since the physicochemical parameters of a new drug candidate are necessary for assessing future PK characteristics and optimizing the candidate, the most important physicochemical parameters of ATB6015, such as solubility, ionization degree (pKa), lipophilicity, and melting temperature, were analyzed.

[0153] (1) Drug solubility is one of the most important physical properties in new drug development. As shown in the table below, the intrinsic solubility of ATB6015 was obtained by the pH-metric method (potentiometric method) in the range of pH 2 to 12, and the result was 94.19 μM, as shown in Table 4 below.

[0154] Intrinsic Sol.T / °CMethodIonic EnvironmentAssay Type94.18 μM24.5pH-metric0.167 M KClCurve fitting

[0155] (2) Ionization degree (pKa) is a property that can predict the physicochemical behavior of a compound and is a parameter that affects lipid content, solubility, protein binding, and permeability. As shown in Table 5 below, the pKa of ATB6015 was confirmed to have two values ​​as a structural characteristic of the compound, with 3.89 and 5.17.

[0156] pKaTypeT / °CIonic EnvironmentMethod3.89 ± 0.05Base24.90.171 M KClpH-metric5.17 ± 0.03Base24.90.171 M KClpH-metric

[0157] (3) Lipophilicity (logP / logD) is a property that is closely related to the absorption of a drug in the human body, and was analyzed by measuring the distribution coefficient in water and octanol. As shown in Table 6 below, the LogP value of ATB6015 was measured as 2.41.

[0158] LogPTypeLogD at pH 7.4 (blood)T / °CIonic EnvironmentMethod2.41 ± 0.03Neutral2.4025.00.169 M KClpH-metric

[0159] (4) Melting point is a unique property of the compound. The melting point was measured using differential scanning calorimetry (DSC), and the values ​​were found to be 163 ℃ - 165 ℃, as shown in Table 7 below.

[0160] Melting point analysis method Analysis conditions Sampling / pretreatment 163℃ - 165℃ DSC Analysis - 10℃ ~ 300℃ range 52 min Standard Sealing Pan (GCA-0052, Aluminum)

[0161]

[0162] Example 3. Development of an optimized candidate material for improving the physical properties of ATB6015.

[0163] Example 3-1. Synthesis of ATB6015 and 5 derivatives and 6 chlorides (salt form)

[0164] In order to find a substance with improved antifungal activity and solubility than the antifungal agent candidate ATB6015, seven derivatives of ATB6015 and five salt compounds thereof were synthesized.

[0165]

[0166] Synthesis method of antifungal drug candidate ATB6015 and 7 derivatives (ATC0001 - ATC0007)

[0167] 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.

[0168]

[0169] Compound 1 was refluxed with a functional group-containing substance 2a-2h, K2CO3 and acetone solvent to synthesize intermediates 3a-3h with functional groups introduced thereto as follows.

[0170]

[0171] Intermediates 3a - 3h with functional groups introduced were stirred at room temperature for 24 hours using 4.0 M HCl in dioxane and EtOAc as a solvent to perform Boc deprotection, thereby synthesizing salt compounds 4a - 4h as follows.

[0172]

[0173] Compound 6 (ATB6015) and 7 derivatives (ATC0001 - ATC0007, compound numbers 6-13) were finally synthesized at room temperature using DMTMM and DMF solvents with functional group-containing substance 5 and different analogues 4a - 4h, as follows.

[0174]

[0175]

[0176] Method for synthesizing chlorides (salt forms) of ATB6015 and its five derivatives (ATC0001 - ATC0005)

[0177] Compounds 6 to 11 synthesized above were finally synthesized at room temperature using 4.0 M in dioxane and EtOAc solvent as follows: Compound 14 (ATB6015 salt), Compound 15 to 19 (5 derivatives, ATC0001 salt - ATC0005 salt).

[0178]

[0179] Through the above compound synthesis method, the antifungal drug candidate ATB6015 and 7 derivatives (ATC0001-0005) and 6 of its chlorides (salt forms) were successfully synthesized. The drug names are as follows. In addition, NMR data is shown in Fig. 7a and below.

[0180] Serial number drug name structure 1ATB6015 2ATC0001 free 3ATC0002 free 4ATC0003 free 5ATC0004 free 6ATC0005 free 7ATC0006 free 8ATC0007 free 9ATB6015 salt 10ATC0001 salt 11ATC0002 salt 12ATC0003 salt 13ATC0004 salt 14ATC0005 salt

[0181]

[0182] Example 3-2. Additional synthesis of five ATB6015 derivatives (ATC0013, ATC0017, ATC0018, ATC0019, ATC0020)

[0183] Compound 20 was synthesized as follows using starting material and 4b as a solvent, NMM, IBCF and THF, by stirring at room temperature for 2 hours.

[0184]

[0185] Compound 20 synthesized above was used as a solvent in 4.0 M dioxane and EtOAc, and compound 21 (ATC0013) was finally synthesized at room temperature as follows.

[0186]

[0187] Compound 23 was synthesized at room temperature using DMTMM and DMF solvents in substances 5 and 22 having functional groups as follows.

[0188]

[0189] Compound 23 synthesized above was synthesized with different analogues 24a - 24d using CS2CO3 and DMF solvents as follows: 120 o Four derivatives 25a - 25d were finally synthesized using a microwave reactor in C.

[0190]

[0191] Through the above compound synthesis method, five additional derivatives (ATC0013, ATC0017-0020) were successfully synthesized, and their drug names are as follows. In addition, NMR data is shown in Figure 7a and below.

[0192] Serial number drug name structure 1ATC0013 2ATC0017 3ATC0018 4ATC0019 5ATC0020

[0193]

[0194] Example 3-3. In vitro activity screening for deriving ATB6015-based optimization candidate substances.

[0195] Using the 13 candidate drugs synthesized based on ATB6015, the minimum inhibitory concentration (MIC) was analyzed to screen for the ability to inhibit drug-resistant Candida strains and invasive strains. Drug activity evaluation was performed on the following fungi:

[0196] - Four invasive fungal species (Candida albicans (SC5314), Candida tropicalis (KCCM 50075), Fusarium oxysporum (KCCM60555), and Aspergillus fumigatus (CEA10))

[0197] - Multidrug-resistant Candida strains showing resistance to antifungal agents (Candida albicans (12-99), Candida glabrata (BG-2))

[0198] The test drug was dissolved in 100% DMSO solvent and treated with the medium at a final ratio of 1% according to the Clinical Standards Institute (CLSI) Guideline Edition 4 M27 (Table 2-1), and the final cell number of C. albicans (SC5314), C. albicans (12-99), C. glabrata (BG-2), and Candida tropicalis (KCCM 50075) was prepared as 500 cells, and the experiment was conducted as shown in Table 10 below.

[0199] RPMI 1640 medium 10.4 gram MOPS buffer 34.53 gram NaOH Adjust to pH 7 Add sterile distilled water to make a total volume of 1 L

[0200] MIC analysis results showed that among the ATB6015 derivatives, ATC0001 free showed lower MIC values ​​overall than ATB6015 against two invasive fungi and two azole-resistant Candida spp. ATC0001 free showed the greatest antifungal activity compared to ATB6015 among the 13 derivatives and was effective against various Candida spp. In particular, the in vitro antifungal activity of ATC0001 free against the azole-resistant strains C. albicans (12-99) and C. glabrata (BG2) was confirmed to be approximately 10- to 32-fold higher than that of ATB6015, as shown in Table 11 below. In addition, ATC0003 free, ATC0003 salt, and ATC0006 free compounds showed the highest antifungal activity against C. albicans (SC5314), C. albicans (12-99), and C. glabrata (BG2). tropicalis (KCCM 50075) strain showed in vitro antifungal activity at a level similar to that of ATB6015, and ATC0004 free showed 2- to 4-fold improved in vitro antifungal efficacy compared to ATB6015 in azole drug-resistant strains C. albicans (12-99) and C. glabrata (BG2).

[0201] DrugMIC (㎍ / mL)C. albicans(SC5314)C. albicans(12-99)C. glabrata(BG-2)C. tropicalis(KCCM 50075)Amphotericin B0.250.250.50.125Fluconazole1>64>64>64ATB60150.25421-4ATC0001 free0.030.1250.250.5-1ATC0002 free8646464ATC0003 free0.5484ATC0004 free0.2511>64ATC0005 free>64>64>64>64ATC0006 free1442ATC0007 free>64>64>64>64ATB6015 salt0.5822ATC0001 salt0.030.1250.25>64ATC0002 salt864644ATC0003 salt0.5481ATC0004 salt0.25>64>64>64ATC0005 salt>64>64>644

[0202] Meanwhile, the compound exhibited antifungal activity not only against Candida spp. but also against filamentous fungi such as Aspergillus spp. and Fusarium spp. As a result of measuring the MIC in Aspergillus fumigatus (CEA10) and Fusarium oxysporum (KCCM60555), as shown in Table 12 below, the antifungal activity of ATC0001 free against the Aspergillus fumigatus strain was improved by about 17 times, and the antifungal activity of ATC0001 free against Fusarium oxysporum was 66 times higher. In addition, the azole-resistant strains C. albicans (12-99) and C. In the case of ATC0004 free, which showed 2 to 4 times improved antifungal efficacy compared to ATB6015 in A. glabrata (BG2), it did not show efficacy against Fusarium oxysporum (KCCM60555), but showed about 4 times improved efficacy against Aspergillus fumigatus (CEA10).

[0203] Compound NameA.fumigatus(CEA10)F.oxysporum(KCCM60555)ATB60150.5∼12ATC0001 free0.03∼0.060.03ATC0002 free4∼84∼8ATC0004 free0.125∼0.5>64ATC0005 free>64>64

[0204] The MIC screening results of ATB6015-based derivatives showed that ATC0001 free has in vitro antifungal activity against invasive Candida fungi, multidrug-resistant Candida, and filamentous fungi, confirming that it is a compound that exhibits a broad-spectrum effect against various strains.

[0205]

[0206] Example 3-4. In vitro activity screening of additionally synthesized derivatives

[0207] In vitro activity screening was performed on five additional compounds (ATC0013, ATC0017, ATC0018, ATC0019, ATC0020) containing aminonicotinamide structures.

[0208] To screen for the ability of five candidate drugs to inhibit representative Candida strains by analyzing the minimal inhibitory concentration (MIC), drug activity evaluation was performed against the following fungi:

[0209] - Two invasive fungal species (Candida albicans (SC5314), Candida auris (B8441))

[0210] The minimum inhibitory concentration (MIC) test developed by the European Committee on Antimicrobial Susceptibility Testing (EUCAST) was performed by dissolving the test drug in 100% DMSO solvent and treating the medium with a final ratio of 2%. The cell concentration corresponding to the OD600 of C. albicans (SC5314) and C. auris (B8441) was used. 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 a 96-well plate containing the diluted drug. The 96-well plate was incubated at 35°C for 2 days, and the cell concentration of each well was measured at OD595. After measurement, the culture solution was spread on YPD medium and cultured at 30°C for 24 hours to confirm the cidal effect.

[0211] As shown in Figs. 6a and 6b, among the five compounds, ATC0017 had the lowest MIC value and was confirmed to have an antifungal effect against two types of invasive fungi. Compounds ATC0018 and ATC0019 showed antifungal effects, but their MIC values ​​were higher than those of ATC0017 and they did not show bactericidal effects. ATC0020 showed antifungal effects only against C. albicans in C. auris, but its MIC values ​​were higher than those of compounds ATC0017, ATC0018, and ATC0019. ATC0013 did not show antifungal effects against either type of invasive fungi. All five additional compounds showed higher MIC values ​​than ATC0001.

[0212]

[0213] Example 3-4. Solubility comparison for deriving optimized candidate materials based on ATB6015.

[0214] In order to evaluate how much the solubility of the five derivatives and salt-form compounds was improved compared to ATB6015, the concentration was adjusted to 2.56 mg / ml, which is the maximum concentration of the drug treated in the MIC experiment, and the degree of solubility in DW and PEG400 solvents, which are widely used as excipients, was evaluated on a 6-point scale, and 100% of each solvent was used (the solubility score was evaluated in 6 levels, from 1 when insoluble to 6 when completely dissolved, and 2 when particles remain and settle or layer separation occurs, 3 when some particles remain but do not settle, 4 when fine particles are visible, and 5 when it is cloudy and completely dissolved or takes a long time to dissolve.)

[0215] 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 forms was improved in DW compared to ATB6015, and the ATB6015 salt form showed the most improved solubility of 6 points in DW. Among the ATB6015 derivatives, ATC0001 free showed a solubility increase of 2 points compared to ATB6015 in 100% DW solvent when the drug was dissolved based on 2.56 mg / ml, and was completely dissolved in 100% PEG400 solvent, showing a solubility score increase of 1 point compared to ATB6015.

[0216] Compound DW solubility PEG400 solubility Compound DW solubility PEG400 solubility ATB601515ATB6015 salt64ATC0001 free36ATC0001 salt25ATC0002 free25ATC0002 salt42ATC0003 free42ATC0003 salt53ATC0004 free35ATC0004 salt25ATC000522ATC0005 salt32

[0217] In summary of the above experiments, ATC0001 free showed about 10-fold to 32-fold improvement in azole-resistant Candida strains compared to ATB6015, and also increased MIC antifungal activity about 17-fold to 66-fold in filamentous strains. To find a compound with improved solubility compared to ATB6015, solubility was measured, and ATC0001 free showed a 2-point increase in solubility in DW and a 1-point increase in solubility in PEG400 compared to ATB6015. Based on the results of broad in vitro antifungal activity and improved solubility in the most commonly used solvents, ATC0001 free was selected as the optimal candidate.

[0218]

[0219] Example 4. Confirmation of the properties of an optimized candidate material based on ATB6015.

[0220] Example 4-1. Physical property evaluation of the optimization candidate material ATC0001 free

[0221] In order to evaluate the physicochemical parameters of ATC0001 free, solubility, ionization degree (pKa), lipophilicity, and melting temperature were measured by a specialized analytical institution.

[0222] (1) As a result of the solubility evaluation, the intrinsic solubility of ATC0001 free was calculated using the pH-metric method (potentiometric method) in the range of pH 2 to pH 12, and the result was 7.923 μM, as shown in the table below.

[0223] Compound NameIntrinsic Sol.T / ℃MethodIonic EnvironmentAssay TypeATC0001 free7.923 μM25.0pH-metric0.168 M KClCheqSol

[0224] (2) Ionization degree (pKa) is a property that can predict the physicochemical behavior of a compound and is a parameter that affects lipid content, solubility, protein binding, and permeability. As shown in the table below, the pKa of ATC0001 free was confirmed to have a value of 4.94.

[0225] Compound NamepKaTypeT / ℃Ionic EnvironmentMethodATC0001 free4.94 ± 0.01Base25.00.168 M KClpH-metric

[0226] (3) Lipophilicity (logP / logD) is a property that is closely related to the absorption of drugs in the human body. It was analyzed by measuring the distribution coefficient in water and octanol, and the LogP value of ATC0001 free was measured as 2.86, as shown below.

[0227] Compound nameLogPTypelogD at pH 7.4 (blood)T / ℃Ionic EnvironmentMethodATC0001 free2.86 ± 0.04Neutral2.8625.00.165 M KClpH-metric

[0228] (4) Melting point is a unique property of the compound. The melting point of ATC0001 free was measured using differential scanning calorimetry (DSC), and the value was confirmed to be 163℃-165℃, as shown in the table below.

[0229] Compound name Melting point Analysis method Analysis conditions Sampling / pretreatment ATC0001 free 167℃ - 169℃ DSC Analysis - 10℃ ~ 300℃ range 52 min Standard Sealing Pan (GCA-0052, Aluminum)

[0230]

[0231] Example 4-2. Comparison of physical properties of ATB6015 and ATC0001 free

[0232] (1) The intrinsic solubility of ATC0001 free was measured to be 7.923 μM, which did not significantly increase the solubility compared to ATB6015. In addition, the solubility was evaluated, and the solubility scores for ATC0001 free in DW and PEG400 increased by 2 and 1 point, respectively, compared to ATB6015, suggesting that ATC0001 free is more suitable as a compound to be dissolved in a solvent for in vivo experiments.

[0233] Compound Name Intrinsic Sol.T / °CMethodIonic EnvironmentAssay TypeATB601594.18 μM24.5pH-metric0.167 M KClCurve fittingATB6015 salt366.6 μM24.3pH-metric0.165 M KClCurve fittingATC0001 free7.923 μM25.0pH-metric0.168 M KClCheqSolATC0001 salt37.89 μM25.0pH-metric0.165 M KClCheqSol

[0234] (2) In addition, due to structural characteristics, two pKa values ​​of ATB6015 and one pKa value of ATC0001 free can be analyzed. As a result of analyzing the ionization degree (pKa) of ATB6015 and ATC0001 free, it was confirmed to have a value of 4.94 as shown in the table below, and it was measured at a similar level to ATB6015, which had pKa values ​​of 3.89 and 5.17.

[0235] Compound NamepKaTypeT / °CIonic EnvironmentMethodATB60153.89 ± 0.05Base24.90.171 M KClpH-metric5.17 ± 0.03Base24.90.171 M KClpH-metricATB6015 salt3.67 ± 0.01Base24.90.161 M KClpH-metric5.12 ± 0.02Base24.90.161 M KClpH-metricATC0001 free4.94 ± 0.01Base25.00.168 M KClpH-metricATC0001 salt4.65 ± 0.04Base25.00.169 M KClpH-metric

[0236] (3) Meanwhile, according to Lipinski's rule, a lipid-based compound must have a LogP value less than 5 to be considered suitable as a drug. As the LogP value of ATB6015 was 2.41 and the free LogP value of ATC0001 was 2.86, both compounds were determined to be suitable for use as drugs, as shown in the table below.

[0237] Compound nameLogPTypelogD at pH 7.4 (blood)T / °CIonic EnvironmentMethodATB60152.41 ± 0.03Neutral2.4025.00.169 M KClpH-metricATB6015 salt2.07 ± 0.03Neutral2.0725.00.171 M KClpH-metricATC0001 free2.86 ± 0.04Neutral2.8625.00.165 M KClpH-metricATC0001 salt1.90 ± 0.06Neutral1.9025.00.169 M KClpH-metric

[0238] (4) Melting point is a unique property of the compound. The melting points of ATB6015 and ATC0001 free were measured using differential scanning calorimetry (DSC). The values ​​were 163℃-165℃ and 167℃-169℃, respectively, and were confirmed to be at similar levels for both compounds, as shown in the table below.

[0239] Compound name Melting point Analysis method Analysis conditions Sampling / Pretreatment 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) ATB0001 free 167℃ - 169℃ DSC Analysis - 10℃ ~ 300℃ range, 52 min Standard Sealing Pan (GCA-0052, Aluminum) ATB0001 salt 158℃ - 160℃ DSC Analysis - 10℃ ~ 300℃ range, 52 min Standard Sealing Pan (GCA-0052, Aluminum)

[0240]

[0241] Example 4-3. Solubility evaluation of optimization candidate ATC0001 free

[0242] To identify excipients used in oral and intravenous animal experiments, the solubility of each solvent was evaluated (the solubility score was evaluated on a 6-level scale, with 1 being insoluble and 6 being completely dissolved. 2 was the level at which particles remained and settled or layer separation occurred, 3 was the level at which some particles remained but did not settle, 4 was the level at which fine particles were visible, and 5 was the level at which the particles were completely dissolved or took a long time to dissolve.).

[0243] ATC0001 was dissolved in various excipients such as PEG, Propylene glycol (PG), Tween, Dimethyl Sulfoxide (DMSO), Carboxy Methyl Cellulose (CMC), and Cyclodextrin (CD) at a concentration of 100 mg / ml to determine the excipient to be used for single-dose oral toxicity administration, and was dissolved at concentrations of 1 to 10 mg / ml to determine the excipient for oral and intravenous injection to be used for efficacy testing.

[0244] When dissolved at a level of 10 mg / ml, it was completely dissolved under the excipient condition of 10% DMSO in corn oil, and when dissolved at a concentration of 6 mg / ml, it was most dissolved when 50% propylene glycol was dissolved in PEG400. When dissolved at a level of 2 mg / ml, it was dissolved in a combination of 5% DMSO, 10% solutol, and 85% PBS, and at a concentration of 1 mg / ml, the solubility was high under the condition of 5% DMSO + 10% solutol in DW or PBS, as shown in the table below. (Table 22).

[0245] Degree of solubility of concentrated active ingredient *100mg / ml100% corn oil350% PEG400 in PG410% DMSO in Corn oil250% PEG400 in Corn oil310mg / ml10% PEG400 in DW130% PEG400 in DW250% PEG400 in DW330% Corn oil in DW250% PG + 2% (Tween 80 1%) in saline350% PG in DW440% CD in water40.5% CMC in water410% DMSO + 40% PEG300 + 5% (Tween 80 1%) in saline310% DMSO in corn oil65% DMSO + 10% PEG400 in CD (40% CD in water)36mg / ml100% corn oil52% CMC in DW45% DMSO + 2% Tween 80 in DW32% Tween 80 in 0.5% CMC35% DMSO + 2% Tween 80 + 40% PEG in 0.5% CMC45% DMSO + 2% Tween 80 in 0.5% CMC410% DMSO in 0.5% CMC3100% olive oil550% Propylene glycol in PEG40064mg / ml5% DMSO + 10% Solutol in PBS42mg / ml10% (50% Castor oil+ 50% EtOH) in DW45% DMSO + 10% Solutol in PBS61mg / ml10% (50% Castor oil+ 50% EtOH) in DW55% DMSO + 10% Solutol in DW or PBS5-6

[0246] Example 5. In vitro activity evaluation of optimization candidate substances against Candida spp.

[0247] Example 5-1. Selection of ATC0001-free optimization candidate and MFC evaluation

[0248] The minimum fungicidal concentration (MFC) of the ATC0001 free candidate substance was measured to determine the fungicidal (fungicidal) / fungistatic (fungal growth inhibition) mode in which the drug kills the strain. For the MFC measurement, after performing the MIC, wells corresponding to 1X, 2X, 4X, and 8X MIC on the MIC plate were plated on a YPD plate, cultured, and colonies were counted to compare the difference with the MIC value. The strains used for the MFC measurement were as follows.

[0249] - Two invasive fungal species (Candida albicans (SC5314), Candida tropicalis (KCCM 50075))

[0250] - Two multidrug-resistant strains (Candida albicans (12-99), Candida glabrata (BG-2))

[0251] As a result of MFC analysis, ATC0001 free was determined to fungistatically inhibit the growth of the invasive fungus Candida albicans (SC5314) and multidrug-resistant strains Candida albicans (12-99) and Candida glabrata (BG-2), similar to ATB6015. In the invasive fungus Candida tropicalis (KCCM 50075), both ATB6015 and ATC0001 free were found to fungicidal kill the strain, and in particular, ATC0001 free showed fungicidal antifungal efficacy even at a lower drug concentration of 4 ㎍ / ml than the MFC value of 16 ㎍ / ml of ATB6015.

[0252] DrugC. albicans(SC5314)C. albicans(12-99)C. glabrata(BG-2)C. tropicalis(KCCM 50075)MICMFCMICMFCMICMFCMICMFCAmphotericin B0.060.125fungicidal0.1250.25fungicidal0.250.25fungicidal0.130.125fungicidalFluconazole2N / A>64N / A16N / A1N / AATB60150.25>2fungistatic4>32fungistatic2>16fungistatic416fungicidalATB6015 salt0.5>2fungistatic8>32fungistatic2>16fungistatic48fungicidalATC0001 free0.03>0.25fungistatic0.131fungistatic0.25>2fungistatic14fungicidal

[0253]

[0254] Example 5-2. Measurement of the biofilm inhibition efficacy of the optimized candidate substance ATC0001 free.

[0255] Fungi form biofilms, which are slimy layers composed of complex proteins, sugars, and lipids, to protect themselves from drugs and external stimuli. Once a biofilm is formed, drug resistance increases approximately 100-fold, making it difficult to effectively eliminate the fungi. When developing antifungal agents, it is important to measure the ability to eliminate fungi that have formed biofilms. The adherence and development inhibition activities of ATB6015 and ATC0001 free were measured in multidrug-resistant Candida fungi, Candida albicans (12-99) and Candida glabrata (BG-2), and the biofilm destruction ability of ATB6015 was compared with that of ATC0001 free.

[0256] The results of measuring the biofilm inhibition activity of ATC0001 free against the strain Candida albicans (12-99) resistant to azole drugs showed that the sessile minimal inhibitory concentration 50 (SMIC) inhibited 50% of the biofilm formed by the strain. 50 ) The activity of inhibiting adherence was increased by more than 512 times compared to ATB6015, and the development inhibition ability was improved by 200 times. The sessile minimal inhibitory concentration 90 (SMIC) that inhibits 90% of biofilm 90 ) ATC0001 free showed a 200-fold increase in biofilm adherence inhibition ability compared to ATB6015, and biofilm development inhibition ability was also improved by 200-fold.

[0257] The results of the biofilm inhibition assay of ATC0001 free on the azole-resistant strain Candida glabrata (BG-2) showed that the sessile minimal inhibitory concentration 50 (SMIC) inhibited 50% of the biofilm formed by the strain. 50 ) showed a 200-fold increase in adherence inhibition activity compared to ATB6015, and also improved the biofilm development inhibition ability by 200-fold. The sessile minimal inhibitory concentration 90 (SMIC) that inhibits 90% of biofilm 90 ) As a result of the measurement, ATC0001 free showed an ability to inhibit biofilm adherence approximately 1,067 times higher than ATB6015, and the degree of inhibition of biofilm development was improved approximately 267 times.

[0258] Candida albicans Biofilm inhibition activity of ATC0001 free (unit: ㎍ / ml) Adherence inhibition SMIC 50Developmentinhibition SMIC 50 Adherence inhibition SMIC 90 Development inhibition SMIC 90 Amphotericin B0.1250.0310.25Fluconazole>6464>64>64ATB6015>642>648ATB6015 salt>642>648ATC0001 free0.125-0.250.01-0.030.250.03-2ATC0001 salt0.125160.030.25

[0259] Candida glabrata (BG-2)에 대한 ATC0001 free의 생물막 저해능 (단위: ㎍ / ml)Adherence inhibition SMIC 50 Developmentinhibition SMIC 50 Adherence inhibition SMIC 90 Development inhibition SMIC 90 Amphotericin B10.06>64>8Fluconazole42>6416ATB601522>648ATB6015 salt22648ATC0001 free0.010.01-0.030.06-0.1250.03-0.06ATC0001 salt0.250.0140.06

[0260] In summary, in the Candida albicans (12-99) strain, ATC0001 free improved the concentration that inhibits 50% of the biofilm by about 200-fold and 512-fold compared to ATB6015 in the adherence and development methods, respectively, and improved 90% inhibition of the biofilm by about 200-fold in both the adherence and development methods. In the Candida glabrata (BG2) strain, the activity of 50% inhibition of the biofilm by both the adherence and development methods increased by 200-fold, and the inhibition of 90% of the biofilm by about 1,067-fold and 267-fold in the adherence and development methods, respectively. Therefore, ATC0001 free has an approximately 200- to 1,067-fold improvement in the ability to inhibit biofilms formed by multidrug-resistant Candida fungi compared to ATB6015.

[0261]

[0262] Example 5-3. Measurement of the emergence of resistant strains of ATC0001 free candidate substances

[0263] Recently, the number of strains showing resistance to antifungal drugs has been increasing, so it is essential to develop antifungal drugs that can effectively inhibit the growth of multidrug-resistant fungi. The MIC test of compounds against the Candida glabrata (ATCC 2001) strain was performed to determine the IC of each drug. 50 The values ​​were determined. Each drug IC 50 Resistance was induced by culturing the fungi on media containing concentrations of 1, 4, 8, 16, and 32X the value. The initial fungal concentration was 10 6 This was inoculated and the number of bacteria increased 100-fold in the medium containing the drug, 10 8 When it reached , it was judged that resistant strains had emerged, and the corresponding date was marked with a circle, and a sample stock was made and stored in a deep freezer. At the same time, 10 new concentration-specific drug media were added. 6The strains were passaged to a number of , and the experiment was repeated daily for 40 days. After 40 days, the measurement of the frequency of emergence of resistant strains was completed, and the MIC (Amphotercin B and ATB6015 derivatives) and IC of the sample stocks were measured. 50 (azole drugs, fluconazole and efinaconazole) were analyzed to determine how much resistance had increased compared to the values ​​before the experiment began. If resistance had increased, the MIC and IC for the same fungal species 50 As the value increases, higher drug concentrations are required to kill the fungus.

[0264] Results of the test to measure the frequency of emergence of resistant strains, IC when ATB6015 free drug was treated in Candida glabrata (ATCC 2001) strain 50 Resistant strains were produced in proportion to the concentration: 15 times at 1X, 13 times at 4X, 10 times at 8X, 7 times at 16X, and 5 times at 32X, and resistant strains appeared on average about 10 times over 40 days. ATC0001 free IC 50 At 1X, it showed 12 times, at 4X, 4 times, at 8X, 3 times, and at 32X, it showed results proportional to the concentration, and resistant strains appeared an average of about 5 times over 40 days. ATC0001 free reduced the frequency of appearance of resistant strains by about twice as much as ATB6015.

[0265] Sample stocks of resistant strains were tested for MIC of three test drugs and commercial amphotericin B, and IC of azole drugs. 50 The results were measured and presented in a graph in Fig. 2.

[0266] The experimental results of the control group were as follows. As a result of the sample analysis of the commercial drug Amphotericin B, it was determined that the resistance to the drug did not increase as the MIC value did not increase in the drug treatment groups of 1, 4, and 8X compared to the MIC value before the experiment. Next, the IC of the azole drug Fluconazole control group was 8 times at 4X, 4 times at 8X, and 8 times at 16X. 50 The values ​​increased and Efinaconazole had a 4X to 4-fold and 8X to 4-fold IC 50 The value has increased.

[0267] In the lead substance ATB6015, the MIC value did not increase in the 1X to 32X groups, and the MIC value of ATB6015 salt increased by 2-fold in 4, 8, and 16X. In the candidate substance ATC0001 free, the MIC value did not increase compared to the pre-experimental analysis value in any group from 1X to 32X. As a result of the analysis of the resistant strain sample stock, it was determined that ATC0001 free did not develop the production of resistant strains because, like the lead substance ATB6015, the MIC value did not increase compared to the pre-experimental value.

[0268] To summarize the contents of Figure 2, in the Candida glabrata (ATCC 2001) strain, the ATC0001 free candidate substance improved the frequency of emergence of resistant strains by two times compared to ATB6015, and the MIC analysis result of the emerged resistant strains showed that ATC0001 free did not induce resistance in the strains, as did ATB6015, since the MIC value did not increase.

[0269]

[0270] Example 6. In vivo activity evaluation of optimized candidate substances against Candida spp.

[0271] Example 6-1. In vivo efficacy evaluation against azole-resistant strain Candida albicans (12-99)

[0272] To establish a systemic fungal infection model, systemic Candida infection was induced in male ICR mice by intravenous administration of Candida albicans (12-99). The antifungal efficacy of commercially available drugs (Caspofungin) and candidate substances (ATB6015, ATC0001 free) was evaluated and compared by oral or intraperitoneal administration. Eight mice were administered per group, and survival rates and body weights were measured for up to 15 days. The excipient composition of the drugs and candidate substances was unified as Castor oil: EtOH: H2O = 5:5:90 (Table 25, Figures 3a and 3b).

[0273] GroupTest ArticlesDoseAdministration amountAdministration routeDosage frequencyDosage periodNumber of individuals / group(mg / kg)(mL / kg)G1Vehicle-5POQD5 days8G2Caspofungin0.1IPG3ATB60155POG4ATB601510POG5ATC0001 free5POG6ATC0001 free10POG7ATC0001 free20PO

[0274] As a result, as shown in Fig. 3a and Fig. 3b, the survival rate and body weight changes of mice after infection (Day 0) were observed for 15 days, and in the control group (G1) without drug administration, the survival rate on the 15th day was 0, meaning all mice died. In contrast, the survival rate on the 15th day of the caspofungin administration group (G2), which was the control group, was 50%, showing the antifungal effect of a general commercial drug. As a result of the experiment, the survival rates on the 15th day of the ATB6015 10 mg / kg administration group (G4) and the ATC0001 free 10 mg / kg administration group (G6) were 62.5%, showing similar results, which was 12.5% ​​higher than the survival rate of caspofungin, which showed a 50% survival rate. Therefore, it was suggested that ATB6015 and ATC0001 free had antifungal efficacy against an animal model of azole-resistant Candida albicans infection.

[0275]

[0276] Example 6-2. In vivo efficacy evaluation against azole-resistant strain Candida albicans (12-99)

[0277] We sought to cross-check the in vivo efficacy against the azole-resistant strain Candida albicans (12-99) by requesting another specialized institution. To establish a fungal systemic infection model, Candida albicans (12-99) was administered intravenously to male ICR mice to induce fungal infection. The antifungal efficacy of commercial antifungal drugs (Caspofungin, Posaconazle) and candidate substances (ATB6015, ATC0001) was evaluated by oral or intraperitoneal administration to the infection model. Survival rates and body weights were compared with 7 mice per group. The composition of the excipients of the drug and candidate substances was unified as Castor oil: EtOH: H2O = 5:5:90 (Table 27, Figures 4a and 4b).

[0278] GroupTest ArticlesDose (mg / kg)Administration amountAdministration routeAdministration frequencyAdministration periodNumber of individuals / group (mL / kg)G1Vehicle-5POQD5 days7G2Caspofungin0.1IPG3Posaconazole2POG4ATB601510POG5ATC0001 free10POG6ATC0001 free20POG7ATC0001 free30POG8ATC0001 free50PO

[0279] As shown in Fig. 4a and Fig. 4b, the body weight and condition of the mice were continuously tracked for 11 days after infection (Day 0). In the case of the control group (G1) without drug administration, the survival rate on Day 11 was 14%, whereas the survival rates on Day 11 of the positive control drug caspofungin administration group (G2) and the negative control drug posaconazole administration group (G3) were 100% and 57%, respectively, showing normal commercial drug antifungal effects.

[0280] In addition, the survival rate on the 11th day of group G5 (10 mg / kg) administered ATB6015 was 86%, and the survival rates of the ATC0001 free 30 mg / kg administered group (G7) and the ATC0001 free 50 mg / kg administered group (G8) were both 100%, showing a 14% increase in survival rate compared to ATB6015. Since the survival rates of ATC0001 free 30 mg / kg and 50 mg / kg were the same as those of the positive control drug caspofungin administered group (G2), it was suggested that ATB6015 and ATC0001 free had antifungal efficacy against an animal model of azole drug-resistant Candida albicans infection.

[0281]

[0282] Example 6-3. In vivo efficacy evaluation against a new multidrug-resistant fungus, Candida auris.

[0283] To establish a systemic fungal infection model, systemic mycosis was induced in male ICR mice by intravenous administration of Candida auris. The antifungal efficacy of commercial antifungal drugs (Caspofungin) and candidate compounds (ATB6015, ATC0001 free) was evaluated by oral or intraperitoneal administration. Eight mice were administered per group, and survival rates and body weights were measured. The excipient composition of the drug and candidate compounds was standardized as Castor oil: EtOH: H2O = 5:5:90 (Table 28, Figures 5a and 5b).

[0284] GroupTest ArticlesDose (mg / kg)Administration amountAdministration routeAdministration frequencyAdministration periodNumber of individuals / group (mL / kg)G1Vehicle-5POQD5 days8G2Caspofungin0.1IPG3ATB601510POG4ATC0001 free10POG5ATC0001 free20POG6ATC0001 free30POG7ATC0001 free40PO

[0285] As shown in Figures 5a and 5b, the body weight and condition of the mice were tracked and observed for 22 days after infection (Day 0). In the case of the control group (G1) without drug administration, the survival rate on Day 22 was 50%, whereas in the control group administered commercially available drugs (G2), the survival rate on Day 22 was 75%, demonstrating the general antifungal effect of commercially available drugs, and the experimental conditions were well-established.

[0286] 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 were 1.5-fold and 1.25-fold improvements, respectively, compared to the survival rate of 50% of the ATB6015 10 mg / kg administration group (G4).

[0287]

[0288] Example 7. PK and stability evaluation of optimization candidate substances

[0289] Example 7-1. PK measurement of ATC0001 free

[0290] Pharmacokinetics (PK) was measured to quantitatively investigate the absorption, distribution, transport, and excretion of the synthetic candidate substance over time after administration in the body. Using SD rats as experimental animals, the candidate substance ATC0001 free was administered orally (per oral, PO) at 10 mg / kg and intravenously (intravenously, IV) at 0.5 mg / kg, and plasma was collected to determine the pharmacokinetics (PK).

[0291] Compound nameRouteDosemg / kg T 1 / 2h T maxh C maxng / mL AUC (0-t)h*ng / mL AUC (0- ∞ ) h*ng / mL MRT (0-t)h MRT (0- ∞)h V zmL / kgClmL / h / kgF%ATB6015IV24.70.082811,0251,0494.95.512,8801,920N / APO1012.942012,4163,4088.318.2N / AN / A47.1ATB6015 saltPO1029.841,75523,46053,6049.841.9N / AN / AN / AATC0001freeIV0.53.50.085601,3401,5103.14.51.725.98N / APO1012.041,51617,84524,3638.717.2N / AN / A66.6ATC0001 saltPO106.241,28514,29115,4357.19.0N / AN / AN / A

[0292] (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)

[0293]

[0294] As shown in Table 29 above, when administered intravenously to SD rats, the maximum concentration (Cmax) value of ATC0001 free was improved by approximately 2-fold 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-fold for ATC0001 free compared to ATB6015.

[0295] As a result of intravenous administration of ATC0001 free, the area under the curve (AUC) value, which indicates the total amount in the blood, was improved by about 1.3 times compared to ATB6015, and about 7.4 times when administered orally. The mean residence time (MRT) in the body was similar for ATC0001 free and ATB6015 when administered intravenously and orally. The bioavailability (F) value, which is an indicator of the amount absorbed into the body after oral administration, was improved by about 1.4 times for ATC0001 free than for ATB6015, indicating increased bioavailability. The bioavailability (F) value, which is an indicator of the amount absorbed into the body after oral administration, was improved by about 1.4 times for ATC0001 free than for ATB6015, indicating increased bioavailability.

[0296] In summary of the above results, it was found that candidate substance ATC0001 free showed significant improvements in most pharmacokinetic indicators, such as maximum blood concentration (Cmax), total blood amount (AUC), and amount absorbed in the body (F), when administered orally and intravenously to experimental animals, compared to ATB6015.

[0297]

[0298] Example 7-2. Stability evaluation results of ATC0001 free

[0299] ATC0001 free was tested by a specialized testing agency to evaluate plasma protein binding, stability in plasma, metabolic stability, hepatocyte stability, and cardiac toxicity, and the stability of the compound was measured as shown in the table below.

[0300] Compound nameSpeciesPlasma protein binding%Plasma stability%Metabolicstability%hepatocyticstabilityCL int ( μL / min) hERGIC 50 (μM)ATB6015Human99.78N / A---Mouse99.82N / A---ATC0001freeHuman98.6>100 (30min)>100 (120min)83.3189.3 (t 1 / 2 )3.7 (CL int )>100Mouse97.4>100 (30min)>100 (120min)78.2427 (t 1 / 2 )1.6 (CL int )

[0301] The degree of binding of ATC0001 free to human and mouse 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, and the result was calculated as a value obtained 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 ​​in the two compounds, with 99.78% in humans and 99.82% in mice.

[0302] In order to evaluate the stability in plasma, the drug was treated and cultured in human and mouse plasma, and the drug was analyzed through 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 levels of values ​​>100% at 30 minutes and 2 hours. Since a plasma stability value of approximately 85% or higher can be evaluated as stable, ATC0001 free can be interpreted as having high stability in vivo.

[0303] 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 a metabolic stability value of 83.3% in humans and 78.2% in mice. Since it is evaluated as a stable compound with a half-life of approximately 1 to 3 hours when the metabolic stability value is 70 to 90%, ATC0001 free could be judged to be a stable drug as a result of the metabolic stability test.

[0304] Additionally, the hepatocyte stability of ATC0001 free was analyzed, and a half-life of 189.3 hours was measured in human hepatocytes and 427 hours in mouse hepatocytes. Clearance criteria (unit: μL / min / 10 6In human hepatocytes, if the clearance value is 3.5 or lower, metabolism is evaluated as slow, and if the value is 19 or higher, metabolism is evaluated as fast. Therefore, ATC0001 free, which showed a value of 3.7, can be interpreted as indicating moderate metabolism in humans. In mouse hepatocytes, if the clearance value is 3.3 or lower, metabolism is evaluated as slow, and if the value is 17.8 or higher, metabolism is evaluated as fast. Therefore, ATC0001 free showed a value of 1.6, and can be evaluated as having a low metabolic rate in mice.

[0305] Next, the hERG cardiotoxicity test was evaluated to assess the arrhythmogenic potential of the drug by measuring the degree of inhibition of the hERG potassium channel activity. In the hERG test, hERG-expressing HEK cells were used without separating human and mouse species, and the IC of ATC0001 free drug was measured. 50 It was evaluated as stable because it had little inhibition of hERG with a value of >100.

[0306]

[0307] In summary, the stability evaluation results of the above ATC0001 free were evaluated by a specialized testing agency to determine that it was stable in terms of stability in plasma and metabolic stability. The results of the hepatocyte stability analysis showed that it had a moderate metabolic rate in human hepatocytes and did not inhibit hERG under the test conditions (100 μM), so it was determined to be a stable compound that did not cause cardiotoxicity.

[0308]

[0309] While specific aspects of the present invention have been described in detail above, it should be apparent to those skilled in the art that these specific descriptions are merely preferred embodiments and do not limit the scope of the present invention. Therefore, the substantial scope of the present invention is defined by the appended claims and their equivalents.

[0310]

[0311] Since the compound of the present invention has a high antifungal effect with low side effects, an antifungal pharmaceutical composition containing the compound is provided.

Claims

1. A compound represented by the following chemical formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the above formula, W is O or S; R1 is hydrogen or C1-6 alkyl; R2 is -OX-R3; X is C1-6 alkyl; R3 is a cycloalkyl group having 5 to 8 carbon atoms, an aryl group, or a monocyclic or bicyclic heteroaryl group containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen and sulfur, and in each case, these groups are unsubstituted or substituted with 1 or 2 halogens, CF 3, may be substituted by an ethoxy group; Y is substituted with one or more amino groups and is heteroaryl or C1 to C containing one or two heteroatoms independently selected from nitrogen, oxygen and sulfur. 10 is an alkylamino group.

2. In paragraph 1, W is O; R1 is hydrogen; X is C1-3 lower alkyl; 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 substituted with one or two halogens, CF 3, can be substituted by a methoxy group; Y is substituted with one amino group and is heteroaryl or C1 to C containing one or two nitrogen heteroatoms. 10 A compound, an isomer or a pharmaceutically acceptable salt thereof, which is an alkylamino group.

3. In paragraph 1, W is O; R1 is hydrogen; X is a methyl group; R3 is a cycloalkyl group, a phenyl group or a pyridyl group having 5 to 8 carbon atoms, and in each case, these groups are unsubstituted or substituted with one or two halogens, CF 3, can be substituted by a methoxy group; A compound, an isomer or a pharmaceutically acceptable salt thereof, wherein Y is a heteroaryl or C1 to C6 alkylamino group substituted with one amino group and containing one nitrogen heteroatom.

4. In paragraph 1, The compound represented by the above chemical formula 1 is a compound selected from the group consisting of compounds represented by the following chemical formulas 2 to 20, an isomer thereof, or a pharmaceutically acceptable salt thereof: [Chemical 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] 5. A pharmaceutical composition for preventing or treating mycosis, comprising a compound according to any one of claims 1 to 4, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

6. In paragraph 5, A pharmaceutical composition having antibacterial activity against bacteria selected from the group consisting of Candida spp., Sporetricum spp., Malassezia spp., Cryptococcus spp., Aspergillus spp., Pneumocystis spp., and Mucor spp.

7. In paragraph 6, A pharmaceutical composition according to claim 1, wherein the mycosis is selected from meningitis, encephalitis due to meningitis, cutaneous candidiasis, mucosal candidiasis, visceral candidiasis, urinary candidiasis, candidal endocarditis, oropharyngeal candidiasis, candidal endophthalmitis, candidal sepsis, tinea versicolor, sporathione, fungal abscess, fungal granuloma, pyogenic granuloma, madura mycosis, Pneumocystis carinii pneumonia, systemic cryptococcosis, mucocutaneous cryptococcosis, aspergillosis, cavities, hemoptysis, allergies, chronic pulmonary tuberculosis, pulmonary fibrosis, pulmonary cysts, chronic fever, cough, sputum, bloody sputum, jock itch, thrush, fungal dementia, and zygomycosis.

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.

9. In paragraph 8, A method having antibacterial activity against bacteria selected from the spp. Candida, Sportrichum, Malassezia, Cryptococcus, Aspergillus, Pneumocystis and Mucor.

10. In paragraph 9, A method according to claim 1, wherein the mycosis is selected from meningitis, encephalitis due to meningitis, cutaneous candidiasis, mucosal candidiasis, visceral candidiasis, urinary candidiasis, candidal endocarditis, oropharyngeal candidiasis, candidal endophthalmitis, candidal sepsis, tinea versicolor, sporathione, fungal abscess, fungal granuloma, pyogenic granuloma, madura mycosis, Pneumocystis carinii pneumonia, systemic cryptococcosis, mucocutaneous cryptococcosis, aspergillosis, cavities, hemoptysis, allergies, chronic pulmonary tuberculosis, pulmonary fibrosis, pulmonary cysts, chronic fever, cough, sputum, bloody sputum, jock itch, thrush, fungal dementia, and zygomycosis.

Images