Novel compound and composition for preventing or treating respiratory diseases containing the same as an active ingredient

Abstract

The present invention relates to a novel compound and a composition for preventing or treating respiratory diseases, which comprises the novel compound, its E- or Z-isomer, its optical isomer, a mixture of two of its isomers, its precursor, a pharmaceutically acceptable salt thereof, or a solvate thereof as an active ingredient.

Description

Technical Field

[0001] The present invention relates to a novel compound and a composition for preventing or treating a respiratory disease containing the same as an active ingredient.

Background Art

[0002] Pendrin is encoded by the SLC26A4 gene, which is an anion exchanger and a member of the SLC26 gene family, and exchanges Cl - for HCO 3 - , I - , OH - and anions such as SCN - . Pendrin is a cell membrane protein expressed on the luminal membrane of airway epithelial cells. However, the expression of pendrin is strongly upregulated in inflammatory airway diseases such as chronic obstructive pulmonary disease (COPD), allergic rhinitis, asthma, Bordetella pertussis infection, acute lung injury (ALI), acute respiratory distress syndrome (ARDS), and common colds caused by rhinovirus, and the upregulation of pendrin is observed when they are cultured with IL-4, IL-13, and IL-17A in primary airway epithelial cells. Interestingly, pendrin knockout (KO) improves airway inflammation in all mouse models of COPD, allergic rhinitis, asthma, Bordetella pertussis infection, and rhinovirus infection. The pathophysiological role of pendrin in airway inflammation has not been clearly elucidated. However, new evidence indicates that pendrin is involved in the regulation of airway surface liquid (ASL) volume preservation and mucus production in inflammatory airway diseases.

[0003] In the culture of primary mouse tracheal epithelial cells with IL-13, the volume increase of ASL was significantly higher in pendrin KO mice compared to the WT mouse control group. In the culture of primary human nasal epithelial (HNE) cells from hearing-impaired patients carrying the pendrin mutant (DFNB4), the IL-13-induced volume increase of ASL was significantly higher than that of the normal control group. Also, the suppression of pendrin by a pendrin inhibitor significantly increased the volume of IL-13-induced ASL in the primary culture of human bronchial epithelial cells. Such findings suggest that the downregulation of pendrin may have a beneficial effect on the regulation of ASL volume homeostasis in inflammatory airway diseases.

[0004] Excessive mucus production is a common feature of inflammatory airway diseases such as asthma and COPD. Overexpression of pendrin significantly increased MUC5AC gene expression in the human lung cancer cell line NCI-H292 and mouse lung tissue. IL-13 treatment significantly increased MUC5AC gene expression in HNE cells from normal subjects, but the IL-13-induced upregulation of MUC5AC was completely abolished in HNE cells from hearing-impaired patients carrying the pendrin mutant. Such research results suggest that the downregulation of pendrin may be beneficial for the treatment of asthma and COPD.

Summary of the Invention

Problems to be Solved by the Invention

[0005] The present invention is based on the discovery that certain compounds can act as pendrin inhibitors that may be useful in treating respiratory diseases. The present invention is based on the discovery that some compounds discovered by cell-based HTS screening for the identification of small molecule pendrin inhibitors or newly designed compounds can provide potential treatments for respiratory diseases (inflammatory airway diseases) such as asthma, acute or chronic bronchitis, allergic rhinitis, acute respiratory infections, acute upper respiratory infections, cystic fibrosis, acute respiratory distress syndrome (ARDS), acute lung injury (ALI), or chronic obstructive pulmonary disease (COPD). The present invention is based on the discovery that some small molecules may be shown to reduce the IL-13-induced upregulation of MUC5AC gene expression in HNE cells differentiated from subjects with normal downregulation of pendrin. Also, some molecules have been used as pendrin inhibitors that improve airway inflammation in a mouse model of ovalbumin (OVA)-induced allergic asthma.

Means for Solving the Problems

[0006] In one aspect, the present invention provides a compound represented by the following Chemical Formula 1, its E- or Z-isomer, its optical isomer, a mixture of two of its isomers, its precursor, its pharmaceutically acceptable salt, or its solvate.

Chem.

Chemical formula

[0007] In another aspect, the present invention provides a pharmaceutical composition for preventing or treating a respiratory disease (inflammatory airway disease), comprising as an active ingredient at least one of the said compounds, at least one of its E- or Z-isomers, at least one of its optical isomers, at least one mixture of two of its isomers, at least one of its precursors, at least one of its pharmaceutically acceptable salts or at least one of its solvates. In some embodiments, the composition can suppress, prevent, improve or treat a respiratory disease (inflammatory airway disease).

[0008] In a further aspect, the present invention provides a composition comprising a compound represented by Chemical Formula 1 or a mixture thereof, and a composition comprising a compound represented by Chemical Formula 1 or a mixture thereof together with a pharmaceutically acceptable carrier.

[0009] In a further aspect, the present invention provides the use of a compound represented by Chemical Formula 1 and its pharmaceutical composition as a pendrin inhibitor.

[0010] In a further aspect, the present invention provides a compound represented by Chemical Formula 1 and its pharmaceutical composition that specifically regulates a chloride channel.

[0011] In a further aspect, the present invention provides a compound represented by Chemical Formula 1 and its pharmaceutical composition that preserves the volume of airway surface liquid (ASL) and reduces the separation of mucin.

[0012] In a further aspect, the present invention provides the use for one or more respiratory diseases (inflammatory airway diseases) selected from the group consisting of asthma, acute or chronic bronchitis, allergic rhinitis, acute respiratory tract infection, acute upper respiratory tract infection, cystic fibrosis, acute respiratory distress syndrome (ARDS), acute lung injury (ALI), and chronic obstructive pulmonary disease (COPD).

[0013] In a further aspect, the present invention provides the use of the compound represented by Chemical Formula 1, its mixture, and its pharmaceutical composition as an active ingredient for the prevention or improvement of respiratory diseases (inflammatory airway diseases) in health functional foods.

[0014] In a further aspect, the present invention provides the use of a compound and its pharmaceutical composition as a pendrin inhibitor as an active ingredient for the prevention or improvement of respiratory diseases (inflammatory airway diseases) in health functional foods.

[0015] In a further aspect, the present invention provides the compound represented by Chemical Formula 1 and its pharmaceutical composition that specifically regulates chloride channels for the prevention or improvement of respiratory diseases (inflammatory airway diseases) as an active ingredient in health functional foods.

[0016] In a further aspect, the present invention provides the compound represented by Chemical Formula 1 and its pharmaceutical composition that preserves the volume of airway surface liquid (ASL) and reduces the separation of mucin for the prevention or improvement of respiratory diseases (inflammatory airway diseases) as an active ingredient in health functional foods.

[0017] In a further aspect, the present invention provides the use as an active ingredient for the prevention or improvement of respiratory diseases (inflammatory airway diseases) in health functional foods, wherein the respiratory diseases (inflammatory airway diseases) are one or more selected from the group consisting of asthma, acute or chronic bronchitis, allergic rhinitis, acute respiratory tract infection, acute upper respiratory tract infection, cystic fibrosis, acute respiratory distress syndrome (ARDS), acute lung injury (ALI), and chronic obstructive pulmonary disease (COPD).

[0018] Other aspects and advantages of the invention will be apparent to those skilled in the art upon consideration of the detailed description and the drawings.

Advantages of the Invention

[0019] According to the present invention, the novel compound can act as a pendrin inhibitor and, as a result, can be usefully used as a composition for the prevention, treatment, or improvement of respiratory diseases (inflammatory airway diseases, particularly asthma or acute lung injury).

Brief Description of the Drawings

[0020]

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BEST MODE FOR CARRYING OUT THE INVENTION

[0021] 1. Definitions Unless otherwise defined, all technical and scientific terms used in the present invention have the meanings commonly understood by one of ordinary skill in the art to which this invention belongs. The following references provide general definitions of many of the terms used in the present invention to those skilled in the art: The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et.al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). As used in the present invention, the following terms have the meanings set forth below unless otherwise specified.

[0022] Unless specifically recited in the context or otherwise apparent, the term "or" used in the present invention is understood to be inclusive.

[0023] Unless specifically recited or apparent from the context, the term "about" as used in this invention is understood within the general tolerances in the art, for example, within two standard deviations of the average. About is understood to be within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05% or 0.01% of the stated value. Unless otherwise clear from the context, all numerical values provided in this invention are modified by the term, about.

[0024] The terms "agent", "drug" and "pharmaceutical formulation" are used interchangeably in this invention to refer to a chemical substance or compound that, when administered to a subject by any means described in this invention (e.g., any animal including a human or non-human animal), induces a desired pharmacological effect (e.g., reduction of inflammation). The "additive" used in the present invention can refer to any additional component added to the compositions and chemical formulas described in the present invention. For example, the additional component provides that it is pharmaceutically acceptable for the particular condition being treated, and the additive can include excipients (e.g., one or more excipients), antioxidants (e.g., one or more antioxidants), stabilizers (e.g., one or more stabilizers), preservatives (e.g., one or more preservatives), pH adjusters and / or buffers (e.g., one or more pH adjusters and / or buffers), isotonicity regulators (e.g., one or more isotonicity regulators), thickeners (e.g., one or more thickeners), suspending agents (e.g., one or more suspending agents), binders (e.g., one or more binders), viscosity increasing agents (e.g., one or more viscosity increasing agents), etc. Also, the additive can include processing agents and drug delivery modifiers, enhancers and any combination of two or more thereof, such as calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starch, gelatin, cellulose, methylcellulose, sodium carboxymethylcellulose, dextrose, hydroxypropyl-beta-cyclodextrin, polyvinylpyrrolidone, low melting point wax, ion exchange resin, etc. Other suitable pharmaceutically acceptable excipients are described in "Remington’s Pharmaceutical Sciences", Mack Pub.Co., New Jersey (1991), and "Remington: The Science and Practice of Pharmacy", Lippincott Williams & Wilkins, Philadelphia, 20th edition (2003) and 21st edition (2005), which are incorporated herein by reference. The additive described in the present invention can be used as any suitable drug.

[0025] As used herein, the term "administer" means to administer to a subject orally, rectally, topically, by intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, intranasal, intravitreal or subcutaneous administration, or by implantation of a sustained release device, e.g., a mini-osmotic pump. Administration can be by any route including parenteral and transmucosal (e.g., oral, nasal, pulmonary, rectal, buccal, vaginal, ocular and transdermal routes).

[0026] As used herein, the terms "analog" and "derivative" are used interchangeably and refer to a compound having the same core as the parent compound, but differing from the parent compound in the order of bonding, in the absence or presence of one or more atoms and / or groups of atoms, and combinations thereof. A derivative may differ from the parent compound, for example, in one or more substituents present on a core that may encompass one or more atoms, functional groups or substructures. Also, a derivative may differ from the parent compound in the order of interatomic bonding within the core. Generally, a derivative can be predicted to be formed from the parent compound by chemical and / or physical steps, at least in theory.

[0027] As used herein, the term "antioxidant" can refer to an artificial or natural substance that prevents and / or delays some types of cell damage and / or oxidation. Antioxidants are found in many foods, including fruits and vegetables. They can also be used as dietary supplements. Exemplary antioxidants can include beta-carotene, lutein, lycopene, selenium, vitamin A, vitamin C and vitamin E. Other antioxidants known to those of skill in the art can also be used. The antioxidants described herein can be used in any suitable amount.

[0028] "Co-administration" means that the compounds or compositions described in the present invention are administered simultaneously immediately before or after the administration of an additional therapeutic or active agent or additive described in the present invention. The compounds or compositions of the present invention may be administered alone or co-administered to a patient. Co-administration is construed to include administering the compounds individually or in combination (one or more compounds or formulations) simultaneously or sequentially. If desired, furthermore, the formulation may be combined with other active substances.

[0029] In the present invention, terms such as "comprising", "comprises", "containing" and "having" can have the meanings belonging to them, and can mean "comprising", "comprises", etc.; similarly, "consisting essentially of" or "consisting essentially of" can also have the meanings belonging to them. The above terms are open-ended and allow the presence of more than what is recited, provided that the basic or novel features of what is recited are not changed by the presence of more than what is recited, except for embodiments of the prior art.

[0030] "Concurrent administration" as used in the present invention includes at least partial overlap in duration. For example, when two formulations (e.g., any formulation or class of formulations described in the present invention having biological activity) are administered concurrently, these administrations occur within a particular desired time period. The administrations of the formulations can start and end on the same day. Also, the administration of one formulation may precede the administration of the second formulation, provided that the two formulations are taken at least once on the same day. Similarly, the administration of one formulation may be extended beyond the administration of the second formulation, provided that the two formulations are taken at least once on the same day. It is not necessary to take the bioactive agent / formulation at the same time every day in order to include concurrent administration.

[0031] The "effective amount" or "therapeutically effective amount" used in the present invention is an amount sufficient to affect a desired biological effect such as a favorable result including clinical outcomes. Thus, the "effective amount" depends on the circumstances to which it is applied. The effective amount varies according to factors known in the art such as the disease state, age, gender, and weight of the individual being treated. Various divided doses may be administered daily, or the dose may be proportionally decreased as indicated by the exigencies of the treatment situation. Also, the compositions / formulations of the present invention may be administered as frequently as necessary to achieve a therapeutic amount.

[0032] The term "gel" used in the present invention can refer to a substance that is not an easily flowing liquid, i.e., a solid, or more precisely, a semi-solid. Gels are formed from natural or synthetic substances. Gels exhibit birefringence and liquid crystal properties with some alignment but not complete alignment. Gels are administrable locally.

[0033] The term "respiratory disease" used in the present invention has its ordinary medical meaning and includes, but is not limited to, asthma, acute or chronic bronchitis, allergic rhinitis, acute respiratory infection, acute upper respiratory infection, cystic fibrosis, acute respiratory distress syndrome (ARDS), acute lung injury (ALI), or chronic obstructive pulmonary disease (COPD), and closely related diseases and disorders of the respiratory system. As used herein, the term "inhibit" means to prevent, reduce, blunt, or stop. In one embodiment, when comparing the amount or rate in the absence of a compound or composition to that in the presence of the compound or composition, if the amount or rate of a process or reaction occurring in the presence of the compound or composition is reduced by at least about 10%, the composition or compound is considered to inhibit the viability of at least one protein (e.g., pendrin). In other embodiments, when comparing the amount or rate in the absence of a compound or composition to that in the presence of the compound or composition, if the amount or rate of a process or reaction occurring in the presence of the compound or composition is reduced by at least about 20%, the composition or compound is considered to inhibit the process or reaction. In other embodiments, when comparing the amount or rate in the absence of a compound or composition to that in the presence of the compound or composition, if the amount or rate of inhibition occurring in the presence of the compound or composition is reduced by about 25% or more, about 30%, about 40%, about 50%, about 60%, about 70%, about 75% or about 80%, the compound or composition is considered to inhibit one or more proteins (e.g., pendrin). In other embodiments, a compound or composition is considered to inhibit the viability of one or more proteins, i.e., to prevent its development.

[0034] As used herein, "intermittent administration" includes a period during which a formulation is administered (which is considered the "first dosing period"), then a period during which the formulation is not ingested or is ingested at a lower dose (which is considered the "off-period"), and then a period during which the formulation is administered again (which is considered the "second dosing period"). Generally, the dosing level of the formulation during the second dosing period is the same as that administered during the first dosing period, but can be increased or decreased as medically necessary. As used herein, "jelly" is a semi-solid system composed of a gel, which is a suspension consisting of small inorganic particles or one of large organic molecules penetrated by a liquid, usually water, in a portion with a high structural cohesiveness matrix.

[0035] The "liquid" used in the present invention is an administration form composed of a composition in a liquid state. The liquid can be poured; it behaves by flowing out of a container at room temperature. The liquid exhibits Newtonian or pseudoplastic flow behavior.

[0036] In an embodiment, the "semi - liquid" used in the present invention can have all the properties of a liquid and other formulations (i.e., suspensions, emulsions, solutions, creams, gels, jellies, etc.).

[0037] The term "ointment" used in the present invention can denote a high - viscosity liquid or semi - liquid dosage form that can be used for the therapeutic treatment of a disease, syndrome or condition.

[0038] The "pharmaceutically acceptable carrier" used in the present invention includes any and all physiologically suitable solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption - delaying agents, etc. The type of carrier can be selected based on the intended route of administration. Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions for the in - situ preparation of sterile topical solutions or dispersions and sterile powders. The use of such media and formulations for pharmaceutically active substances is well known in the art. The use thereof in the compositions for the present invention is contemplated, provided that any conventional medium or formulation is not incompatible with the composition (e.g., chemical formula 1, derivatives or analogs of chemical formula 1, or pharmaceutically acceptable salts, solvents, hydrates or polymorphs thereof as described in the present invention).

[0039] The "pharmaceutical carrier" or "carrier" used in the present invention can additionally include a pharmaceutically acceptable carrier, excipient or stabilizer that is non-toxic to cells or mammals at the employed dosage and concentration. Physiologically acceptable carriers are sometimes aqueous pH buffer solutions. Examples of physiologically acceptable carriers are buffers such as phosphates, citrates and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates including glucose, mannose or dextrin; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as Tween TM , polyethylene glycol (PEG) and Pluronics TM . Additionally, "pharmaceutically acceptable" means approved or approvable by a regulatory agency of the Federal or State government or in a country other than the United States by the corresponding agency and listed in the U.S. Pharmacopeia or other generally recognized pharmacopeias for use in animals and more particularly in humans.

[0040] The term "pharmaceutically acceptable salt or complex" refers to salts or complexes of the compound represented by the following specified Chemical Formula 1. Examples of such salts include hydroxides, carbonates or bicarbonates of metal cations such as those selected from the group consisting of alkali metals (e.g., sodium, potassium or lithium) and alkaline earth metals (e.g., calcium or magnesium), or organic or inorganic bases, or base addition salts formed by the reaction of the compound represented by Chemical Formula 1 with a primary, secondary, or tertiary alkylamine, including but not limited to amines derived from methylamine, dimethylamine, trimethylamine, ethylamine, diethylamine, triethylamine, morpholine, N-methyl-D-glucamine, N,N'-bis(phenylmethyl)-1,2-ethanediamine, tromethamine, ethanolamine, diethanolamine, ethylenediamine, N-methylmorpholine, procaine, piperidine, piperazine, etc., which are considered to be within the scope of the present invention.

[0041] Also, the "salt" or "salt form" or "pharmaceutically acceptable salt" used in the present invention can include, for example, base addition salts (formed with free carboxyl or other anionic groups) derived from inorganic bases such as sodium, potassium, ammonium, calcium or ferric hydroxide, and organic bases such as isopropylamine, trimethylamine, 2-ethylamino-ethanol, histidine, procaine, etc. Such salts can be formed as acid addition salts with any free cationic group, for example, generally formed with inorganic acids such as hydrochloric acid, sulfuric acid or phosphoric acid, or organic acids such as acetic acid, citric acid, p-toluenesulfonic acid, methanesulfonic acid, oxalic acid, tartaric acid, mandelic acid, etc. The salts of the present invention can include amine salts formed by protonation of amino groups with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, etc. Also, the salts of the present invention include amine salts formed by protonation of amino groups with suitable organic acids such as p-toluenesulfonic acid, acetic acid, etc.

[0042] The terms "pH formulation" or "buffer" as used in the present invention can refer to compounds useful as pH adjusters or buffers. These can include, but are not limited to, glycerol buffers, citrate buffers, borate buffers, acetate buffers, gluconate buffers, phosphate buffers or citrate-phosphate buffers. The pH formulation or buffer can be used in any suitable amount.

[0043] The term "preservative" as described in the present invention can refer to a substance or chemical that prevents undesirable chemical changes in the compounds or compositions or chemical formulas described in the present invention. Suitable preservatives are, for example, benzalkonium chloride, thimerosal, chlorobutanol, methylparaben, propylparaben, phenylethyl alcohol, disodium edetate, sorbic acid, onamer M polyquat, cetyl bromide, cetylpyridinium chloride, benzyl bromide, EDTA, phenylmercury nitrate, phenylmercury acetate, thimerosal, merthiolate, acetate and phenylmercury borate, polymyxin B sulfate, methyl and propylparaben, quaternary ammonium chloride, sodium benzoate, sodium propionate and sodium perborate, and other formulations known to those skilled in the art, or combinations thereof. The preservative can be used in any suitable amount.

[0044] The terms "prevent", "preventing" or "prevention" and other grammatical equivalents as used in the present invention include those for preventing not only the occurrence of a syndrome but also the development, occurrence, interference or avoidance of a disease or condition syndrome. Prevention can be complete (i.e., without detectable symptoms) or partial, with fewer symptoms observed than when there is no treatment. The term additionally includes prophylactic benefits. To prevent a disease or condition, the composition is administered to a patient at risk of developing a particular disease or a patient who reports one or more physiological syndromes of such a disease but may not have a diagnosis of such a disease.

[0045] The ranges provided in the present invention are understood to be shorthand for all values within the range. For example, a range of 1 to 10 includes not only all intermediate decimal values between the aforementioned integers such as 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, and 1.9, but also any number, combination of numbers, or sub-range from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In relation to sub-ranges, "overlapping sub-ranges" extending from one of the endpoints of the range are specifically considered. For example, the overlapping sub-ranges of an exemplary range of 1 to 50 can include 1 to 10, 1 to 20, 1 to 30, and 1 to 40 in one direction, or 50 to 40, 50 to 30, 50 to 20, and 50 to 10 in the other direction. A range is expressed in the present invention as "about" one particular value and / or "about" another particular value. When such a range is expressed, other aspects include one particular value and / or another particular value. Similarly, when a value is expressed as an approximation using the preceding "about", it is understood that the particular value forms other aspects. It is further understood that each endpoint of a range is important in relation to, and independent of, the other endpoint. Also, there are a number of values disclosed in the present invention, and it is understood that each value is also disclosed in the present invention as "about" that particular value in addition to the value itself. Also, throughout the application, data is provided in a number of different formats, and such data is understood to indicate endpoints, starting points, and ranges for any combination of data points. For example, if a particular data point "10" and a particular data point "15" are disclosed, it is considered that not only between 10 and 15, but also 10 and above 15, 10 and below 15, and equivalents are disclosed. Further, it is understood that each unit between two particular units is disclosed. For example, if 10 and 15 are disclosed, 11, 12, 13, and 14 are also disclosed.

[0046] Additional excipients contemplated for use in the examples of the present invention are those available to those skilled in the art, for example, as found in United States Pharmacopoeia Vol. XXII and National Formulary Vol. XVII, U.S. Pharmacopoeia Convention, Inc., Rockville, Md. (1989), the relevant content of which is incorporated herein by reference.

[0047] The "semi-solid gel" according to the present invention is semi-solid. The apparent viscosity of the semi-solid preparation can increase according to the concentration.

[0048] The "sequential administration" used in the present invention includes that the administration of two formulations (for example, the compounds or compositions described in the present invention) occurs individually on the same day or does not occur on the same day (for example, occurs on consecutive days).

[0049] The "solution" according to the present invention may be a transparent and homogeneous liquid dosage form containing one or more chemical substances dissolved in a solvent or a mixture of solvents that are miscible with each other. A solution is a liquid preparation containing one or more dissolved chemical substances in a suitable solvent or a mixture of solvents that are miscible with each other. Since the molecules of the drug substance in the solution are uniformly dispersed, the use of the solution as a dosage form generally provides assurance of a uniform dosage amount at the time of administration and good accuracy when the solution is diluted or mixed with others.

[0050] The term "solvent" used in the present invention refers to a liquid solvent that is aqueous or non-aqueous. The choice of solvent depends particularly on the solubility of the composition in the solvent and the mode of administration. The aqueous solvent may be composed of only water or composed of water and one or more miscible solvents, and can contain dissolved solutes such as sugars, buffers, salts or other excipients. More commonly used non-aqueous solvents are short-chain organic alcohols such as methanol, ethanol, propanol, short-chain ketones such as acetone, and polyalcohols such as glycerol. "Subject" or "patient" means a human or non-human animal such as a mammal. The "subject" can include any animal including horses, dogs, cats, pigs, goats, rabbits, hamsters, monkeys, guinea pigs, rats, mice, lizards, snakes, sheep, cows, fish and birds. A human subject can refer to a patient.

[0051] The "suspension" used in the present invention is a liquid dosage form containing solid particles dispersed in a liquid vehicle.

[0052] The "viscosity" used in the present invention refers to the flow resistance of a fluid. Viscosity agents can be used in the present invention and include, for example, polyvinyl alcohol, polyvinylpyrrolidone, methylcellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, carboxymethylcellulose, hydroxypropylcellulose, other formulations known to those skilled in the art, or combinations thereof.

[0053] The term "weight percentage" or " %(w / w)" refers to the percentage of a component in a solution calculated based on the weights of the component and the solvent. For example, a 1% (w / w) solution of a component would have 1 g of the component dissolved in 100 g of the solvent. The term "volume percentage" or " %(v / v)" refers to the percentage of a component in a solution calculated based on the volumes of the component and the solvent. For example, a 1% (v / v) solution of a component would have 1 ml of the component dissolved in 100 ml of the solvent. The term "weight / volume percentage" or " %(w / v)" refers to the percentage of a component in a solution calculated based on the weight of the component and the volume of the solvent. For example, a 1.0% (w / v) solution of a component would have 1 g of the component dissolved in 100 ml of the solvent.

[0054] As used herein, the term "syndrome" refers to a condition characterized by a group of symptoms that occur persistently together or a series of related symptoms. A syndrome (e.g., acute respiratory distress syndrome) can be a series of medical signs and symptoms that are related to each other and sometimes associated with a specific disease. In contrast, a disease can be a health condition with a subsequently clearly defined cause. However, a syndrome (derived from the Greek word meaning "running together") can induce a number of symptoms without a confirmable cause. They can imply the possibility of an underlying disease or the occurrence of a disease.

[0055] As used herein, the terms "treat", "treating", or "treatment", and other grammatical equivalents, include the alleviation, weakening, improvement, or prevention of a disease, condition (e.g., acute respiratory distress syndrome), or symptoms, the prevention of additional symptoms, the improvement or prevention of the underlying metabolic cause of the symptoms, the suppression of a disease or condition, e.g., the arrest of the development of a disease or condition, the alleviation of a disease or condition, the regression of a disease or condition, the alleviation of a condition caused by a disease or condition, or the cessation of the symptoms of a disease or condition, and are intended to include prevention. The terms additionally include achieving a therapeutic benefit and / or a prophylactic benefit. A therapeutic benefit means the eradication or improvement of the underlying disorder being treated. Also, a therapeutic benefit is achieved by the eradication or improvement of one or more physiological symptoms associated with the underlying disorder, such that an improvement is observed in the patient, even though the patient still suffers from the underlying disorder.

[0056] The term "functional food" refers to a food or food supplement manufactured or processed as a raw material, functional ingredient, active pharmaceutical ingredient, or additive useful for improving and / or nourishing and / or preserving the physiological functions of the human body.

[0057] The term "acute respiratory distress syndrome (ARDS)" refers to a medical condition that occurs in critically ill patients with widespread inflammation in the lungs. ARDS is a clinical phenotype that can be caused by various pathologies such as pneumonia and sepsis. Widespread damage to cells that form the alveolar barrier, surfactant dysfunction, abnormal coagulation, and activation of the innate immune response are characteristic of ARDS.

[0058] The term "acute lung injury (ALI)" refers to an inflammatory syndrome and increased permeability associated with hypoxemia and a classic radiographic appearance. The most severe end of such a spectrum is ARDS.

[0059] The term "airway surface liquid (ASL)" refers to a thin layer of fluid that coats the apical surface of the airway epithelium at the air interface. ASL plays a central role in maintaining airway homeostasis. The volume, pH, and ionic balance of ASL are directly involved in the regulation of antibacterial activity, ciliary function, and mucociliary clearance.

[0060] The term "inflammatory airway disease" refers to a variety of inflammatory airway disorders including asthma, acute or chronic bronchitis, allergic rhinitis, acute respiratory infections, acute upper respiratory infections, cystic fibrosis, acute respiratory distress syndrome (ARDS), acute lung injury (ALI), chronic obstructive pulmonary disease (COPD), and the like.

[0061] As used in the context of the present invention, the term "inhibitor" is defined as a molecule, two or more molecules, or a pharmaceutical composition that completely or partially inhibits the activity of a target or two or more targets that induce a desired biological effect. Non-limiting exemplary targets include enzymes, receptors, ion-channels, or transporters (e.g., pendrin), etc. An "inhibitor" can inhibit a target reversibly or irreversibly, and reversible inhibition includes competitive inhibition, uncompetitive inhibition, non-competitive inhibition, and mixed inhibition. The term "alkyl", when used alone or in combination with other terms, denotes a straight-chain or branched-chain C monovalent alkyl group having 1 to 20 carbon atoms 1 -C 20It contains alkyl. Such terms are exemplified as a group such as methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, t-butyl, n-pentyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, tetrahydrogeranyl, n-dodecyl, n-tridecyl, n-tetradecyl, n-pentadecyl, n-hexadecyl, n-octadecyl, n-nonadecyl and n-eicosanyl. Preferably, these are C 1 -C 9 alkyl, more preferably C 1 -C 6 alkyl, particularly preferably C 1 -C 4 alkyl, which similarly represents a monovalent alkyl group having 1 to 9 carbon atoms, a monovalent alkyl group having 1 to 6 carbon atoms, and a monovalent alkyl group having 1 to 4 carbon atoms, respectively.

[0062] The term "alkenyl", when used alone or in combination with other terms, refers to a linear or branched C 2 -C 20It contains alkenyl. It may have any available number of double bonds at any available position, and the configuration of the double bond may be (E) or (Z). Such terms are exemplified as a group such as vinyl, allyl, isopropenyl, 1-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-ethyl-1-butenyl, 3-methyl-2-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 4-methyl-3-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-heptenyl, 1-octenyl, geranyl, 1-decenyl, 1-tetradecenyl, 1-octadecenyl, 9-octadecenyl, 1-eicosenyl and 3,7,11,15-tetramethyl-1-hexadecenyl. Preferably, these are C 2 -C 8 alkenyl, more preferably C 2 -C 6 containing alkenyl. Among them, vinyl or ethenyl (-CH=CH 2 ), n-2-propenyl (allyl, -CH 2 CH=CH 2 ), isopropenyl, 1-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, 3-methyl-2-butenyl and the like are particularly preferred.

[0063] The term "alkynyl", when used alone or in combination with other terms, refers to straight-chain or branched-chain C 2 -C 20 containing alkynyl. It may have any available number of triple bonds at any available position. Such terms are exemplified as groups such as ethynyl (-C≡1-propynyl, 2-propynyl (propargyl: -CH 2 C≡2-butynyl, 2-penten-4-ynyl and the like, alkynyl groups having 2 to 20 carbon atoms and optionally having double or triple bonds. In particular, these are C 2 -C 8 alkynyl, more preferably C 2 -C 6It contains alkynyl or the like. Preferably, it represents a group having 2 to 6 carbon atoms and having alkynyl unsaturation at at least one or two positions, C 2 -C 6 It contains alkynyl.

[0064] The term "heteroalkyl" refers to C 1 -C 12 -alkyl, preferably C 1 -C 6 -alkyl, wherein at least one of said carbons is replaced by a heteroatom selected from O, N or S, including 2-methoxyethyl or the like.

[0065] The term "aryl" refers to an unsaturated aromatic carbocyclic group of 6 to 14 carbon atoms having a single ring (e.g., phenyl) or a multiple condensed ring (e.g., indenyl, naphthyl, 2,3-dihydro-1H-indenyl, 1,2,3,4-tetrahydronaphthyl). Aryl includes phenyl, naphthyl, anthryl, phenanthrenyl and the like.

[0066] The term "C 1 -C 6 alkylaryl" refers to an aryl group having a C 1 -C 6 alkyl substituent, including methylphenyl, ethylphenyl, t-butylphenyl and the like.

[0067] The term "arylC 1 -C 6 alkyl" refers to a C 1 -C 6 alkyl group having an aryl substituent, including 3-phenylpropanil, benzyl and the like.

[0068] The term "heteroaryl" refers to a monocyclic heteroaromatic, or bicyclic or tricyclic fused-ring heteroaromatic group. Specific exemplifications of heteroaromatic groups include optionally substituted pyridyl, pyrrolyl, pyrimidinyl, furyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, 1H-pyrazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,3,4-triazinyl, 1,2,3-triazinyl, benzofuryl, [2,3-dihydro]benzofuryl, isobenzofuryl, benzothienyl, benzotriazolyl, isobenzothienyl, indolyl, isoindolyl, 3H-indolyl, benzimidazolyl, imidazo[1,2-a]pyridyl, benzothiazolyl, benzoxazolyl, quinolidinyl, quinazolinyl, phthalazinyl, quinoxalinyl, cinnolinyl, naphthyridinyl, pyrido[3,4-b]pyridyl, pyrido[3,2-b]pyridyl, pyrido[4,3-b]pyridyl, quinolyl, isoquinolyl, tetrazolyl, 5,6,7,8-tetrahydroquinolyl, 5,6,7,8-tetrahydroisoquinolyl, purinyl, pteridinyl, carbazolyl, xanthenyl, benzoquinolyl, benzo[d][1,3]dioxol-5-yl, 3,4-dihydro-1H-pyrano[4,3-c]pyridyl, quinolin-2(1H)-one, 4H-chromene, 1H-indole, and the like.

[0069] The term "C 1 -C 6 alkylheteroaryl" refers to a heteroaryl group having a C 1 -C 6 alkyl substituent, including methylfuryl, t-butylfuryl, and the like.

[0070] The term "heteroarylC 1 -C 6 alkyl" refers to a C 1 -C 6 alkyl group having a heteroaryl substituent, including furylmethyl, and the like.

[0071] The term "C 2 -C 6 alkenylaryl" refers to an aryl group having a C 2 -C 6 alkenyl substituent, including vinylphenyl and the like.

[0072] The term "arylC 2 -C 6 alkenyl" refers to a C 2 -C 6 alkenyl group having an aryl substituent, including phenylvinyl and the like.

[0073] The term "C 2 -C 6 alkenylheteroaryl" refers to a heteroaryl group having a C 2 -C 6 alkenyl substituent, including vinylpyridinyl and the like.

[0074] The term "heteroarylC 2 -C 6 alkenyl" refers to a C 1 -C 6 alkenyl group having a heteroaryl substituent, including pyridinylvinyl and the like.

[0075] The term "C 3 -C 8 -cycloalkyl" refers to a saturated carbocyclic group having 3 to 8 carbon atoms with a single ring (e.g., cyclohexyl) or multiple fused rings (e.g., norbornyl). C 3 -C 8 -cycloalkyl includes cyclopentyl, cyclohexyl, norbornyl, and the like.

[0076] The term "heterocycloalkyl" refers to a C 3 -C 8-Refers to cycloalkyl or polycondensed rings. Heterocycloalkyl includes lactam or lactone. Non-limiting examples are pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, decahydroisoquinolinyl, octahydro-1H-pyrano[3,4-c]pyridinyl, 4-methylene-5(4H)-one, pyrrolidin-2-one, etc.

[0077] The term "C 1 -C 6 alkyl C 3 -C 8 -cycloalkyl" refers to C including methylcyclopentyl, etc. 1 -C 6 -cycloalkyl group with an alkyl substituent. 3 -C 8

[0078] The term "C 3 -C 8 -cycloalkyl C 1 -C 6 -alkyl" refers to C including 3-cyclopentylpropyl, etc. 3 -C 8 -alkyl group with a cycloalkyl substituent. 1 -C 6

[0079] The term "C 1 -C 6 -alkyl heterocycloalkyl" refers to C including 4-methylpiperidinyl, etc. 1 -C 6 -heterocycloalkyl group with an alkyl substituent.

[0080] The term "heterocycloalkyl C 1 -C 6 -alkyl" refers to C with a heterocycloalkyl substituent including (1-methylpiperidin-4-yl)methyl, etc. 1 -C 6 -alkyl group.

[0081] The term "carboxy" refers to the group -C(O)OH.

[0082] The term "carboxy C" 1 -C 6 "alkyl" refers to a C 1 -C 6 alkyl group having a carboxy substituent such as 2-carboxyethyl.

[0083] The term "acyl" refers to a group -C(O)R including acetyl etc., where R is H, "alkyl", preferably "C" 1 -C 6 "alkyl", "aryl", "heteroaryl", "C" 3 -C 8 "cycloalkyl", "heterocycloalkyl", "aryl C" 1 -C 6 "alkyl", "heteroaryl C" 1 -C 6 "alkyl", "C" 3 -C 8 "cycloalkyl C" 1 -C 6 "alkyl" or "heterocycloalkyl C" 1 -C 6 "alkyl".

[0084] The term "acyl C" 1 -C 6 "alkyl" refers to a C 1 -C 6 alkyl group having an acyl substituent such as 2-acetylethyl.

[0085] The term "acylaryl" refers to an aryl group having an acyl substituent such as 2-acetylphenyl.

[0086] The term "acyloxy" refers to a group -OC(O)R including acetyloxy etc., where R is H, "C" 1 -C 6 "alkyl", "C" 2 -C 6 "alkenyl", "C" 2 -C 6 "alkynyl", "C" 3 -C 8-Cycloalkyl」, 「heterocycloalkyl」, 「aryl」, 「heteroaryl」, 「aryl C 1 -C 6 -alkyl」, 「heteroaryl C 1 -C 6 -alkyl」, 「aryl C 2 -C 6 -alkenyl」, 「heteroaryl C 2 -C 6 -alkenyl」, 「aryl C 2 -C 6 -alkynyl」, 「heteroaryl C 2 -C 6 -alkynyl」, 「C 3 -C 8 -cycloalkyl C 1 -C 6 -alkyl」 or 「heterocycloalkyl C 1 -C 6 -alkyl」.

[0087] The term 「acyloxy C 1 -C 6 -alkyl」 refers to a C 1 -C 6 -alkyl group having an acyloxy substituent such as 2-(ethylcarbonyloxy)ethyl.

[0088] The term 「alkoxy」 refers to the group -OR, where R is 「C 1 -C 6 -alkyl」, 「aryl」, 「heteroaryl」, 「aryl C 1 -C 6 -alkyl」 or 「heteroaryl C 1 -C 6 -alkyl」. Preferred alkoxy groups include, for example, methoxy, ethoxy, phenoxy, etc.

[0089] The term 「alkoxy C 1 -C 6 -alkyl」 refers to a C 1 -C 6 -alkyl group having an alkoxy substituent such as methoxyethyl.

[0090] The term "alkoxycarbonyl" refers to the group -C(O)OR, where R is "C 1 -C 6 alkyl", "aryl", "heteroaryl", "arylC 1 -C 6 alkyl", "heteroarylC 1 -C 6 alkyl" or "heteroalkyl".

[0091] The term "alkoxycarbonylC 1 -C 6 alkyl" refers to a C 1 -C 6 alkyl group having an alkoxycarbonyl substituent such as 2-(benzyloxycarbonyl)ethyl.

[0092] The term "aminocarbonyl" refers to the group -C(O)NRR' including N-phenylcarbonyl, etc., where R and R' are independently H, C 1 -C 6 alkyl, aryl, heteroaryl, "arylC 1 -C 6 alkyl" or "heteroarylC 1 -C 6 alkyl".

[0093] The term "aminocarbonylC 1 -C 6 alkyl" refers to an alkyl group having an aminocarbonyl substituent including 2-(dimethylaminocarbonyl)ethyl, N-ethylacetamidyl, N,N-diethyl-acetamidyl, etc.

[0094] The term "acylamino" refers to the group -NRC(O)R' including acetylamino, etc., where R and R' are independently H, "C 1 -C 6 alkyl" "C 2 -C 6 alkenyl" "C 2 -C 6 alkynyl" "C 3 -C 8-Cycloalkyl", "heterocycloalkyl", "aryl", "heteroaryl", "aryl C 1 -C 6 alkyl", "heteroaryl C 1 -C 6 alkyl", "aryl C 2 -C 6 alkenyl", "heteroaryl C 2 -C 6 alkenyl", "aryl C 2 -C 6 alkynyl", "heteroaryl C 2 -C 6 alkynyl", "cycloalkyl C 1 -C 6 alkyl" or "heterocycloalkyl C 1 -C 6 alkyl". The term "acylamino C 1 -C 6 alkyl" includes a C 1 -C 6 alkyl group having an acylamino substituent such as 2-(propionylamino)ethyl.

[0095] The term "ureido" refers to the group -NRC(O)NR’R’’, where R, R’ and R’’ are independently H, "C 1 -C 6 alkyl", "alkenyl", "alkynyl", "C 3 -C 8 cycloalkyl", "heterocycloalkyl", "C 1 -C 6 aryl", "heteroaryl", "aryl C 1 -C 6 alkyl", "heteroaryl C 1 -C 6 alkyl", "aryl C 2 -C 6 alkenyl", "heteroaryl C 2 -C 6 alkenyl", "aryl C 2 -C 6 alkynyl", "heteroaryl C 2 -C 6"alkynyl", "cycloalkyl C" 1 -C 6 "alkyl" or "heterocycloalkyl C" 1 -C 6 "alkyl", and at this time, R' and R'' may optionally form a 3- to 8-membered heterocycloalkyl ring together with the nitrogen atom to which they are attached.

[0096] The term "ureido C" 1 -C 6 "alkyl" refers to a C-alkyl group having a ureido substituent such as 2-(N'-methylureido)ethyl. 1 -C 6 alkyl group.

[0097] The term "carbamate" refers to the group -NRC(O)OR', and at this time, R and R' are independently "C" 1 -C 6 "alkyl", "C" 2 -C 6 "alkenyl", "C" 2 -C 6 "alkynyl", "C" 3 -C 8 "-cycloalkyl", "heterocycloalkyl", "aryl", "heteroaryl", "C" 1 -C 6 "alkylaryl", "heteroaryl C" 1 -C 6 "alkyl", "aryl C" 2 -C 6 "alkenyl", "heteroaryl C" 2 -C 6 "alkenyl", "aryl C" 2 -C 6 "alkynyl", "heteroaryl C" 2 -C 6 "alkynyl", "cycloalkyl C" 1 -C 6 "alkyl" or "heterocycloalkyl C" 1 -C 6 "alkyl", and also, R may be hydrogen.

[0098] The term "amino" refers to the group -NRR', where R and R' are independently H, "C 1 -C 6 -alkyl", "aryl", "heteroaryl", "C 1 -C 6 -alkylaryl", "C 1 -C 6 -alkylheteroaryl", "cycloalkyl" or "heterocycloalkyl", and where R and R' may optionally form, together with the nitrogen atom to which they are attached, a 3- to 8-membered heterocycloalkyl ring.

[0099] The term "aminoalkyl" refers to an alkyl group having an amino substituent, including, for example, 2-(1-pyrrolidinyl)ethyl.

[0100] The term "ammonium" refers to the positively charged group -N + RR'R'', where R, R' and R'' are independently "C 1 -C 6 -alkyl", "C 1 -C 6 -alkylaryl", "C 1 -C 6 -alkylheteroaryl", "cycloalkyl" or "heterocycloalkyl", and where R and R' may optionally form, together with the nitrogen atom to which they are attached, a 3- to 8-membered heterocycloalkyl ring.

[0101] The term "ammoniumalkyl" refers to an alkyl group having an ammonium substituent, including, for example, 1-ethylpyrrolidinium.

[0102] The term "halogen" refers to fluoro, chloro, bromo and iodo atoms.

[0103] The term "sulfonyloxy" refers to the group -OSO 2 R, where R is "C 1 -C 6 -alkyl" or "C 1 -C 6 -alkyl" substituted with halogen, for example, -OSO2 CF 3 group, "C 2 -C 6 -alkenyl", "-alkynyl", "C 3 -C 8 -cycloalkyl", "-heterocycloalkyl", "aryl", "heteroaryl", "arylC 1 -C 6 -alkyl", "heteroarylC 1 -C 6 -alkyl", "arylC 2 -C 6 -alkenyl", "heteroarylC 2 -C 6 -alkenyl", "arylC 2 -C 6 -alkynyl", "heteroarylC 2 -C 6 -alkynyl", "cycloalkylC 1 -C 6 -alkyl" or "-heterocycloalkylalkyl" selected therefrom.

[0104] The term "sulfonyloxyC 1 -C 6 -alkyl" refers to an alkyl group having a sulfonyloxy substituent including, for example, 2-(methylsulfonyloxy)ethyl.

[0105] The term "sulfonyl" refers to the group "-SO 2 R", where R is "aryl", "heteroaryl", "C 1 -C 6 -alkyl" substituted with halogen, "C 1 -C 6 -alkyl", for example, -SO 2 CF 3 group, "C 2 -C 6 -alkenyl", "C 2 -C 6 -alkynyl", "C 3 -C 8 -cycloalkyl", "-heterocycloalkyl", "aryl", "heteroaryl", "arylC 1 -C 6 -alkyl", "heteroarylC 1 -C6 "alkyl", "aryl C" 2 -C 6 "alkenyl", "heteroaryl C" 2 -C 6 "alkenyl", "aryl C" 2 -C 6 "alkynyl", "heteroaryl C" 2 -C 6 "alkynyl", "cycloalkyl C" 1 -C 6 "alkyl" or "heterocycloalkyl C" 1 -C 6 is selected from "alkyl".

[0106] The term "sulfonyl C" 1 -C 6 "alkyl" refers to an alkyl group having a sulfonyl substituent such as 2-(methylsulfonyl)ethyl.

[0107] The term "sulfinyl" refers to the group "-S(O)R", where R is "alkyl" substituted with "alkyl" halogens, for example, -SOCF 3 group, "C" 2 -C 6 "alkenyl", "C" 2 -C 6 "alkynyl", "C" 3 -C 8 "cycloalkyl", "heterocycloalkyl", "aryl", "heteroaryl", "aryl C" 1 -C 6 "alkyl", "heteroaryl C" 1 -C 6 "alkyl", "aryl C" 2 -C 6 "alkenyl", "heteroaryl C" 2 -C 6 "alkenyl", "aryl C" 2 -C 6 "alkynyl", "heteroaryl C" 2 -C 6 "alkynyl", "C" 3 -C 8 -cycloalkyl C 1 -C 6"alkyl" or "heterocycloalkyl C" 1 -C 6 is selected from "alkyl".

[0108] The term "sulfinylalkyl" refers to an alkyl group having a sulfinyl substituent including, for example, 2-(methylsulfinyl)ethyl.

[0109] The term "sulfanyl" refers to the group -SR, where R is H, "C" 1 -C 6 alkyl", "C" substituted with halogen 1 -C 6 alkyl", e.g., -SCF 3 group, "C" 2 -C 6 alkenyl", "C" 2 -C 6 alkynyl", "C" 3 -C 8 -cycloalkyl", "heterocycloalkyl", "aryl", "heteroaryl", "aryl C" 1 -C 6 alkyl", "heteroaryl C" 1 -C 6 alkyl", "aryl C" 2 -C 6 alkenyl", "heteroaryl C" 2 -C 6 alkenyl", "aryl C" 2 -C 6 alkynyl", "alkynyl heteroaryl", "cycloalkyl C" 1 -C 6 alkyl" or "heterocycloalkyl C" 1 -C 6 alkyl" and includes. Preferred sulfanyl groups include methylsulfanyl, ethylsulfanyl, and the like.

[0110] The term "sulfanyl C" 1 -C 6 alkyl" refers to a C" having a sulfanyl substituent including, for example, 2-(ethylsulfanyl)ethyl 1 -C 5 -alkyl group.

[0111] The term "sulfonylamino" refers to the group -NRSO 2 R’, where R and R’ are independently "C 1 -C 6 alkyl", "C 2 -C 6 alkenyl", "C 2 -C 6 alkynyl", "C 3 -C 8 -cycloalkyl", "heterocycloalkyl", "aryl", "heteroaryl", "arylC 1 -C 6 alkyl", "heteroarylC 1 -C 6 alkyl", "arylC 2 -C 6 alkenyl", "heteroarylC 2 -C 6 alkenyl", "arylC 2 -C 6 alkynyl", "heteroarylC 2 -C 6 alkynyl", "C 3 -C 8 cycloalkylC 1 -C 6 alkyl" or "heterocycloalkylC 1 -C 6 alkyl".

[0112] The term "sulfonylaminoC 1 -C 6 alkyl" refers to an alkyl group having a sulfonylamino substituent including 2-(ethylsulfonylamino)ethyl and the like.

[0113] The term "aminosulfonyl" refers to the group -SO 2 NRR’, where R and R’ are independently H, "C 1 -C 6 alkyl", "C 2 -C 6 alkenyl", "C 2 -C 6 alkynyl", "C 3 -C 8-Cycloalkyl", "heterocycloalkyl", "aryl", "heteroaryl", "arylC 1 -C 6 -alkyl", "heteroarylC 1 -C 6 -alkyl", "arylalkenyl", "heteroarylC 2 -C 6 -alkenyl", "arylC 2 -C 6 -alkynyl", "heteroarylC 2 -C 6 -alkynyl", "C 3 -C 8 -cycloalkylC 1 -C 6 -alkyl" or "heterocycloalkylC 1 -C 6 -alkyl", and at this time, R and R' may optionally form a 3- to 8-membered heterocycloalkyl ring together with the nitrogen atom to which they are attached. The aminosulfonyl group includes cyclohexylaminosulfonyl, piperidinylsulfonyl, and the like.

[0114] The term "aminosulfonylC 1 -C 6 -alkyl" includes a C 1 -C 6 -alkyl group having an aminosulfonyl substituent such as 2-(cyclohexylaminosulfonyl)ethyl.

[0115] Unless otherwise restricted by the definition of individual substituents, all of the above substituents must be understood to be optionally substituted.

[0116] Unless otherwise restricted by the definition of individual substituents, the term "substituted" means "C 1 -C 6 -alkyl", "C 2 -C 6 -alkenyl", "C 2 -C 6 -alkynyl", "C 3 -C 8 -cycloalkyl", "heterocycloalkyl", "C 1 -C6 "alkylaryl", "C" 1 -C 6 "alkylheteroaryl", "C" 1 -C 6 "alkylcycloalkyl", "C" 1 -C 6 "alkylheterocycloalkyl", "amino", "aminosulfonyl", "ammonium", "acylamino", "aminocarbonyl", "aryl", "heteroaryl", "sulfinyl", "sulfonyl", "alkoxy", "alkoxycarbonyl", "carbamate", "sulfanyl", "halogen", "trihalomethyl", "cyano", "hydroxy", "mercapto", "nitro", etc., and refers to a group substituted with 1 to 5 substituents selected from the group consisting of these.

[0117] 2. Compound One aspect of the present invention provides a compound represented by the following Chemical Formula 1, its E- or Z-isomer, its optical isomer, a mixture of two of its isomers, its precursor, its pharmaceutically acceptable salt or its solvate.

Chemical formula

[0118] Non-limiting examples of the compounds include the following compounds described in Tables 1 and 2.

[0119]

Table 1-1

Table 1-2

Table 1-3

Table 1-4

Table 1-5

Table 1-6

Table 1-7

Table 1-8

Table 1-9

[0120]

Table 2-1

Table 2-2

Table 2-3

[0121] The term "compound of the present invention" and equivalent expressions in this specification include the compound represented by Chemical Formula 1 described above. Such expressions include its E- or Z-isomers, its optical isomers, a mixture of two of its isomers, its precursors, its pharmaceutically acceptable salts or its solvates, and are newly synthesized.

[0122] The present invention is further illustrated by the following examples which do not limit the scope of the present invention in any way.

[0123] 3. Manufacturing method Another aspect of the present invention provides a method for manufacturing a compound represented by Chemical Formula 1. The synthesis method is well described in the detailed examples described in the present invention.

[0124] In some embodiments, during organic synthesis, the use of a base may be an organic or inorganic base. Non-limiting examples of the organic base include pyridine, trimethylamine, N,N-diisopropylethylamine (DIPEA), and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). Non-limiting examples of the inorganic base include sodium hydroxide, sodium carbonate, potassium carbonate, cesium carbonate, and sodium hydride. These can be used stoichiometrically or in excess, either alone or in combination. Non-limiting examples of solvents that can be used include ethers (e.g., tetrahydrofuran (THF), diethyl ether, and 1,2-dimethoxyethane), alcohols (e.g., methanol, ethanol, propanol, and butanol), dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dichloromethane (DCM), dichloroethane, water, and acetone. The solvents can be used alone or in combination.

[0125] 4. Composition / Dosage form The present invention includes a pharmaceutical composition comprising the compounds described in the present invention and a dosage form suitable for administration of the compounds described in the present invention. The present invention includes dosage forms of pharmaceutical compositions suitable for administration by any medically acceptable means. The pharmaceutical dosage form can include pharmaceutically acceptable additives or carriers and pharmaceutically acceptable compounds (compositions) suitable for the means of administration.

[0126] The compounds described in the present invention may be in dosage forms (including pharmaceutical compositions) having additives such as excipients (e.g., one or more excipients), antioxidants (e.g., one or more antioxidants), stabilizers (e.g., one or more stabilizers), preservatives (e.g., one or more preservatives), pH adjusters and / or buffers (e.g., one or more pH adjusters and / or buffers), isotonicity regulators (e.g., one or more isotonicity regulators), thickeners (e.g., one or more thickeners), suspending agents (e.g., one or more suspending agents), binders (e.g., one or more binders), viscosity increasing agents (e.g., one or more viscosity increasing agents), etc., and are provided as pharmaceutically acceptable additional components for the particular condition to be treated. In some embodiments, the dosage form can include a combination of additional components (e.g., 2, 3, 4, 5, 6, 7, 8 or more additional components) as described in the present invention. In some embodiments, the additives can include, for example, calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starch, gelatin, cellulose, methylcellulose, sodium carboxymethylcellulose, dextrose, hydroxypropyl-beta-cyclodextrin, polyvinylpyrrolidone, low melting point wax, ion exchange resin, etc., treatment agents and drug delivery modifiers, enhancers and any combination of two or more thereof.

[0127] Other suitable pharmaceutically acceptable excipients are described in "Remington’s Pharmaceutical Sciences", Mack Pub.Co., New Jersey (1991), and "Remington: The Science and Practice of Pharmacy", Lippincott Williams & Wilkins, Philadelphia, 20th edition (2003) and 21st edition (2005), which are incorporated herein by reference.

[0128] The dosage forms of the compositions described in the present invention can be suitable for oral administration in the form of inhalation, nasal spray, intravenous, intramuscular injection, intravitreal injection, as an ointment or in solutions, suspensions, semi - liquids, semi - solids, gels, semi - solid gels, jellies, emulsions, ointments, tablets, liquids and creams. Tablet forms can include one or more of lactose, sucrose, mannitol, sorbitol, calcium phosphate, corn starch, potato starch, microcrystalline cellulose, gelatin, colloidal silicon dioxide, talc, magnesium stearate, stearic acid and other excipients, colorants, fillers, binders, diluents, buffers, humectants, preservatives, flavoring agents, dyes, disintegrants and pharmaceutically suitable carriers. Capsules can contain the compound together with suitable excipients or the compound can be used alone in the shell. All of these formulated compounds can be administered alone, co - administered, intermittently administered, sequentially or simultaneously.

[0129] 5. Administration The compositions of the present invention include, but are not limited to, those administered by any mode of oral, parenteral, sublingual, transdermal, rectal, transmucosal, topical, inhalation, buccal or intranasal administration or combinations thereof. Parenteral administration includes, but is not limited to, intravenous, intra - arterial, intraperitoneal, subcutaneous, intramuscular, intrathecal and intra - arterial. Also, the compositions of the present invention may be administered by implant, which allows for not only sustained release of the composition but also controlled - release intravenous administration.

[0130] The dosage administered to an individual in single or multiple doses will vary widely depending on various factors including pharmacokinetic properties, the condition and characteristics of the patient (gender, age, weight, health, size), the degree of the symptoms, concurrent therapy, frequency of treatment and the desired effect.

[0131] According to one embodiment of the present invention, the compounds according to the present invention and their pharmaceutical dosage forms can be administered alone or together with adjuvants useful in the treatment of respiratory disorders or diseases. According to other embodiments of the present invention, the compounds according to the present invention and their pharmaceutical dosage forms may be administered together with radiotherapy.

[0132] The present invention includes the administration of a compound according to the present invention or a pharmaceutical dosage form thereof, which is administered to an individual in a therapeutically effective amount prior to, simultaneously with, or sequentially with other therapeutic therapies or adjuvants (e.g., multi-drug therapy) useful for the treatment of cancer. The compound according to the present invention or its pharmaceutical dosage form administered simultaneously with the adjuvant can be administered in the same or different compositions and by the same or different routes of administration.

[0133] In one embodiment, the patient according to the present invention is a patient suffering from a respiratory disorder or disease such as bronchial asthma, bronchitis, allergic rhinitis, adult respiratory distress syndrome, cystic fibrosis, pulmonary viral infection (influenza), pulmonary hypertension, idiopathic pulmonary fibrosis, and chronic obstructive pulmonary disease (COPD).

[0134] 6. Use according to the present invention In other embodiments, the present invention provides the use of a compound represented by Chemical Formula 1, a mixture of compounds, or a pharmaceutical composition thereof for the prevention, improvement, or treatment of a respiratory disease (inflammatory airway disease).

[0135] In other embodiments, the present invention provides the use of a compound represented by Chemical Formula 1 and a pharmaceutical composition thereof as a pendrin inhibitor.

[0136] In other embodiments, there is provided the use of a compound represented by Chemical Formula 1 and a pharmaceutical composition thereof, which preserves the volume of airway surface liquid (ASL) and reduces the secretion of mucin.

[0137] In other embodiments, the present invention provides the use for a respiratory disease (inflammatory airway disease) selected from one or more of asthma, acute or chronic bronchitis, allergic rhinitis, acute respiratory infection, acute upper respiratory infection, cystic fibrosis, acute respiratory distress syndrome (ARDS), acute lung injury (ALI), or chronic obstructive pulmonary disease (COPD).

[0138] In other embodiments, the present invention provides the use of a compound represented by Chemical Formula 1, a mixture of compounds, or a pharmaceutical composition thereof as an active ingredient in a health functional food for the prevention or improvement of respiratory diseases (inflammatory airway diseases).

[0139] In other embodiments, the present invention provides the use of a compound represented by Chemical Formula 1 and a pharmaceutical composition thereof as a pendrin inhibitor as an active ingredient in a health functional food for the prevention or improvement of respiratory diseases (inflammatory airway diseases).

[0140] In other embodiments, the present invention provides the use of a compound represented by Chemical Formula 1 and a pharmaceutical composition thereof, which specifically regulates chloride channels as an active ingredient in a health functional food for the prevention or improvement of respiratory diseases (inflammatory airway diseases).

[0141] In other embodiments, the present invention provides the use of a compound represented by Chemical Formula 1 and a pharmaceutical composition thereof, which preserves the volume of airway surface liquid (ASL) and reduces the secretion of mucus as an active ingredient in a health functional food for the prevention or improvement of respiratory diseases (inflammatory airway diseases).

[0142] In other embodiments, the present invention provides the use as an active ingredient in a health functional food for the prevention or improvement of respiratory diseases (inflammatory airway diseases), wherein the respiratory diseases (inflammatory airway diseases) are selected from one or more of the group consisting of asthma, acute or chronic bronchitis, allergic rhinitis, acute respiratory infection, acute upper respiratory infection, cystic fibrosis, acute respiratory distress syndrome (ARDS), acute lung injury (ALI), or chronic obstructive pulmonary disease (COPD).

Example

[0143] [Example] Non-limiting examples of detailed experiments that do not limit the overall experiments are described in the present invention. The description of the present invention described herein serves as an example, and those having general knowledge of the technology related to the present invention should understand that it can be easily changed to other specific areas or forms without changing the technical idea and essential features of the present invention. The described present invention presents the following examples illustratively, but is not limited thereto.

[0144] The names of the compounds were generated by ChemDraw Professional V.15.1. The compounds according to the present invention include not only the compounds represented by Chemical Formula 1, their tautomers, their geometric isomers (e.g., e, z isomers), their optically active forms as optical isomers, their diastereomers, and their racemic forms, but also their pharmaceutically acceptable salts. The derivatives exemplified in the present invention can be prepared from readily available starting materials using the following general methods and procedures. Given typical or preferred experimental conditions (i.e., reaction temperature, time, molar amount of reagents, solvent, etc.), it will be understood that other experimental conditions can be used unless otherwise mentioned. The optimal reaction conditions vary depending on the specific reactants or solvents used, but such conditions can be determined by those skilled in the art using routine optimization procedures.

[0145] The references cited in the present invention are incorporated herein by reference in their entirety. The present invention is not limited in scope to the specific embodiments described herein, which are intended as single illustrations of individual aspects of the present invention, and functionally equivalent methods and components are within the scope of the present invention. Indeed, various modifications of the present invention will become apparent to those skilled in the art from the foregoing description and the accompanying drawings. Such modifications are intended to fall within the scope of the appended claims.

[0146] Example 1-2: Preparation of New Compounds A synthesized compound can exist as an isomer or a mixture of isomers due to the nature of synthetic chemistry or their inherent physical / chemical properties. In some cases, it is interconvertible with other isomers. For example, the compound "F1" means (E)-4-(thiophen-2-ylmethylene)-2-(4-(trifluoromethyl)phenyl)oxazol-5(4H)-one, (Z)-4-(thiophen-2-ylmethylene)-2-(4-(trifluoromethyl)phenyl)oxazol-5(4H)-one, or a mixture of the two isomers. Such phenomena are well documented by the study of Graziano et al (Tetrahedron 62 (2006) 1165 - 1170).

[0147] Example 1.1: 4-(Thiophen-2-ylmethylene)-2-(4-(trifluoromethyl)phenyl)oxazol-5(4H)-one (F1)

Chemical Structure

[0148] Glycine (180 mg, 2.40 mmol) and 4-(trifluoromethyl)benzoyl chloride (0.411 ml, 2.758 mmol) were sequentially added to a 10% NaOH solution (24 ml) at room temperature under stirring, and the mixture was stirred at such a temperature for 2 hours. After acidification to pH 2 with HCl aq. solution, the resulting solid was filtered through a filtering funnel, and the solid was washed with methylene chloride to obtain 2-(4-(trifluoromethyl)benzamido)acetic acid. Such 2-(4-(trifluoromethyl)benzamido)acetic acid (529 mg, 2.14 mmol) and 1-ethyl-3-(3-dimethylamino)propylcarbodiimide hydrochloride (410 mg, 2.14 mmol) were sequentially added to methylene chloride (21 ml) at room temperature under stirring. Then, 1H-pyrrole-2-carbaldehyde (200 mg, 1.783 mmol) and triethylamine (0.497 ml, 3.567 mmol) were added at room temperature under stirring, and the mixture was stirred at such a temperature for 12 hours. The solvent was evaporated under reduced pressure. The resulting solid was filtered through a filtering funnel and washed with methanol to provide the desired product (4-(thiophen-2-ylmethylene)-2-(4-(trifluoromethyl)phenyl)oxazol-5(4H)-one, F1).

[0149] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.22 (d, 2H, J = 8.4 Hz), 8.08 (d, 1H, J = 5.2 Hz), 7.96 (d, 2H, J = 8.4 Hz), 7.86 (d, 1H, J = 3.6 Hz), 7.79 (s, 1H), 7.24 (t, 1H, J = 4.2 Hz)

[0150] ESI (m / z) 324 (MH + )

[0151] Example 1.2: 2-(4-(tert-Butyl)phenyl)-4-(thiazol-5-ylmethylene)oxazol-5(4H)-one (F2)

Chemical Structure

[0152] Thionyl chloride (2.038 ml, 28.05 mmol) and dimethylformamide (10 drops) were successively added to 4-tert-butylbenzoic acid (500 mg, 2.81 mmol) in methylene chloride under stirring, and the mixture was heated for 12 hours. The solvent was evaporated to obtain an intermediate (0.542 ml, 2.758 mmol). Glycine (180 mg, 2.398 mmol) and such an intermediate were successively added to 10% NaOH aq. solution (24 ml) at room temperature under stirring, and the mixture was stirred at such a temperature for 2 hours. After acidifying to pH 2 with aq. HCl solution, the resulting solid was filtered through a filtering funnel and washed with methylene chloride to obtain 2-(4-(tert-butyl)benzamido)acetic acid. 2-(4-(tert-butyl)benzamido)acetic acid (500 mg, 2.125 mmol) and 1-ethyl-3-(3-dimethylamino)propylcarbodiimide hydrochloride (448 mg, 2.338 mmol) were successively added to methylene chloride (21 ml) at room temperature under stirring. Thiazole-5-carbaldehyde (216 mg, 1.913 mmol) and triethylamine (0.519 ml, 3.70 mmol) were successively added to such a mixture at room temperature under stirring, and the whole mixture was stirred for 12 hours. The solvent was removed under reduced pressure. The resulting solid was obtained through a filtering funnel, and the solid was washed with methanol to provide the desired product (2-(4-(tert-butyl)phenyl)-4-(thiazol-5-ylmethylene)oxazol-5(4H)-one, F2).

[0153] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.15 (d, 1H, J = 2.8 Hz), 8.10 (d, 1H, J = 2.8 Hz), 8.03 (d, 2H, J = 8.4 Hz), 7.65 (d, 2H, J = 8.4 Hz), 7.40 (s, 1H), 1.30 (s, 9H)

[0154] ESI (m / z) 313 (MH + )

[0155] Example 1.3: 2-(4-(tert-Butyl)phenyl)-4-((2-methyl-1H-indol-3-yl)methylene)oxazol-5(4H)-one (F3) [Chemical formula]

[0156] Thionyl chloride (2.04 ml, 28.05 mmol) and dimethylformamide (10 drops) were sequentially added to a methylene chloride solution of 4-tert-butylbenzoic acid (500 mg, 2.81 mmol) at room temperature under stirring, and the mixture was heated for 12 hours. The solvent was removed under reduced pressure to give an intermediate (0.542 ml, 2.758 mmol). Glycine (180 mg, 2.398 mmol) and such an intermediate were sequentially added to a 10% NaOH aq. solution (24 ml) at room temperature under stirring. The whole mixture was stirred at room temperature for 2 hours. After acidifying with HCl until pH 2 was reached, the resulting solid was filtered through a filter funnel, washed with methylene chloride, to give 2-(4-(tert-butyl)benzamido)acetic acid. 2-(4-(tert-Butyl)benzamido)acetic acid (400 mg, 2.073 mmol) and 1-ethyl-3-(3-dimethylamino)propylcarbodiimide hydrochloride (433.54 mg, 2.262 mmol) were sequentially added to methylene chloride (20 ml) at room temperature under stirring. 2-Methyl-1H-indole-3-carbaldehyde (300 mg, 1.885 mmol) and triethylamine (0.315 ml, 2.262 mmol) were sequentially added to such a mixture at room temperature under stirring, and the whole mixture was stirred for 12 hours. The solvent was removed under reduced pressure, and the resulting solid was filtered through a filter funnel. The solid was washed with methanol to give the desired product (2-(4-(tert-butyl)phenyl)-4-((2-methyl-1H-indol-3-yl)methylene)oxazol-5(4H)-one, F3).

[0157] 1 1H NMR (400 MHz, MHz, DMSO-d 6) δ 12.25 (s, 1H), 9.16 (d, 1H, J = 7.9 Hz), 7.96 (d, 2H, J = 8.4 Hz), 7.61 (d, 2H, J = 8.0 Hz), 7.37 (s, 2H), 7.25 - 7.18 (m, 2H), 2.62 (s, 3H), 1.30 (s, 9H)

[0158] ESI (m / z) 359 (MH + )、357 (MH - )

[0159] Example 1.4: tert-Butyl 3-((2-(4-(tert-butyl)phenyl)-5-oxooxazol-4(5H)-ylidene)methyl)-1H-indole-1-carboxylate (F4) [Chemical formula]

[0160] To a stirred solution of indole-5-carbaldehyde (200 mg, 1.38 mmol) in 4 mL of acetonitrile was added di-tert-butyl dicarbonate (361 mg, 1.65 mmol), followed by 4-dimethylaminopyridine (DMAP; 17 mg, 0.14 mmol). The mixture was stirred at room temperature for 2 hours. Water (20 mL) was added and the product was extracted with dichloromethane (3 × 20 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (ethyl acetate:hexane = 1:5) to give tert-butyl 3-formyl-1H-indole-1-carboxylate (Step 1).

[0161] Thionyl chloride (2.0 ml, 28.05 mmol) and dimethylformamide (10 drops) were sequentially added to a 0.1 M methylene chloride solution of 4-tert-butylbenzoic acid (500 mg, 2.81 mmol) under stirring. The mixture was heated for 12 hours, and the solvent was removed under reduced pressure to obtain the intermediate chloride (Step 2). Glycine (180 mg, 2.398 mmol) and the intermediate chloride (0.542 ml, 2.758 mmol) were sequentially added to a 10% NaOH solution (23.98 ml) at room temperature under reduced pressure, and the mixture was stirred for 2 hours. The mixture was acidified with HCl until pH 2 was reached, and the solid formed was passed through a filtration funnel. The solid was washed with methylene chloride to afford 2-(4-(tert-butyl)benzamido)acetic acid (Step 3). 2-(4-(tert-butyl)benzamido)acetic acid (200 mg, 0.85 mmol) and 1-ethyl-3-(3-dimethylamino)propylcarbodiimide hydrochloride (179.25 mg, 0.935 mmol) were sequentially added to methylene chloride (8.5 ml) at room temperature under stirring. tert-Butyl 3-formyl-1H-indole-1-carboxylate (208 mg, 0.085 mmol) and triethylamine (0.208 ml, 2.550 mmol) were sequentially added to such a mixture at room temperature under stirring, and the whole mixture was stirred for 12 hours. The solvent was removed at room temperature under reduced pressure, and the solid formed was obtained by passing through a filtration funnel. The solid obtained was washed with methanol to afford the desired product (tert-butyl 3-((2-(4-(tert-butyl)phenyl)-5-oxooxazol-4(5H)-ylidene)methyl)-1H-indole-1-carboxylate, F4).

[0162] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.84 (s, 1H), 8.31 (d, J = 7.6 Hz, 1H), 8.09 (d, J = 7.9 Hz, 1H), 7.99 (d, J = 8.4 Hz, 2H), 7.67 - 7.60 (m, 3H), 7.45 - 7.33 (m, 2H), 1.67 (s, 9H), 1.31 (s, 9H)

[0163] Example 1.5: 4-(5-Oxo-4-(thiophen-2-ylmethylene)-4,5-dihydrooxazol-2-yl)benzonitrile (F5)

Chem.

[0164] Glycine (180 mg, 2.40 mmol) and 4-cyanobenzoyl chloride (460 mg, 2.76 mmol) were successively added to a 10% NaOH solution (24 ml) at room temperature with stirring, and the mixture was stirred at such temperature for 2 hours. After acidifying with HCl until the mixture reached pH 2, the resulting solid (2-(4-cyanobenzamido)acetic acid) was filtered through a filter funnel and then washed with methylene chloride. 2-(4-Cyanobenzamido)acetic acid (400 mg, 1.96 mmol) and 1-ethyl-3-(3-dimethylamino)propylcarbodiimide hydrochloride (410 mg, 2.14 mmol) were successively added to methylene chloride (20 ml) at room temperature with stirring, and the mixture was stirred for 12 hours. Thiophene-2-carbaldehyde (200 mg, 1.783 mmol) and triethylamine (0.5 ml, 3.57 mmol) were successively added to such a solution at room temperature with stirring, and the whole mixture was stirred for 12 hours. The solvent was removed under reduced pressure, and the resulting solid was obtained by filtration through a filter funnel. The solid was washed with methanol to give the desired product (4-(5-oxo-4-(thiophen-2-ylmethylene)-4,5-dihydrooxazol-2-yl)benzonitrile, F5).

[0165] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.18 (d, 2H, J = 8.4 Hz), 8.07 (t, 3H, J = 7.6 Hz), 7.87 (d, 1H, J = 3.6 Hz), 7.81 (s, 1H), 7.25 (t, 1H, J = 4.4 Hz)

[0166] Example 1.6: 2-(4-(tert-Butyl)phenyl)-4-((1-methyl-1H-pyrrol-2-yl)methylene)oxazol-5(4H)-one (F6) [Chemical formula]

[0167] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 7.95 (d, 2H, J = 8.4 Hz), 7.58 (d, 3H, J = 8.0 Hz), 7.24 (s, 1H), 7.17 (s, 1H), 6.33 (t, 1H, J = 3.0 Hz), 3.78 (s, 3H), 1.29 (s, 9H)

[0168] ESI (m / z) 309 (MH + )

[0169] Example 1.7: Methyl 4-(5-oxo-4-(thiophen-2-ylmethylene)-4,5-dihydrooxazol-2-yl)benzoate (F7) [Chemical formula]

[0170] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.16 (q, 4H, J = 8.8 Hz), 8.08 (d, 1H, J = 5.2 Hz), 8.02 (s, 1H), 7.86 (d, 1H, J = 3.6 Hz), 7.79 (s, 1H), 3.88 (s, 3H)

[0171] ESI (m / z) 314 (MH + )

[0172] Example 1.8: 2-(4-(tert-Butyl)phenyl)-4-((4-methylthiazol-5-yl)methylene)oxazol-5(4H)-one (F8) [Chemical formula]

[0173] 1 H NMR (400 MHz, MHz, DMSO-d 6)δ 9.28 (s, 1H), 7.95 (d, 2H, J = 8.4 Hz), 7.62 (d, 2H, J = 8.4 Hz), 7.53 (s, 1H), 2.59 (s, 3H), 1.29 (s, 9H)

[0174] ESI (m / z) 327 (MH + )

[0175] Example 1.9: 2-(4-(tert-Butyl)phenyl)-4-(thiophen-3-ylmethylene)oxazol-5(4H)-one (F9)

Chem.

[0176] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.44 (d, 1H, J = 2.4 Hz), 8.01 - 7.99 (m, 3H), 7.71 - 7.69 (m, 1H), 7.59 (d, 2H, J = 8.8 Hz), 7.37 (s, 1H), 1.29 (s, 9H)

[0177] ESI (m / z) 312 (MH + )

[0178] Example 1.10: 4-((1H-Pyrrol-2-yl)methylene)-2-(4-(tert-butyl)phenyl)oxazol-5(4H)-one (F10)

Chem.

[0179] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 11.53 (s, 1H), 8.08 (d, 2H, J = 8.2 Hz), 7.58 (d, 2H, J = 8.3 Hz), 7.32 (s, 1H), 7.19 (s, 1H), 7.06 (s, 1H), 6.34 (s, 1H), 1.29 (s, 9H)

[0180] ESI (m / z) 295 (MH+ )

[0181] Example 1.11: 4-(Benzo[b]thiophen-2-ylmethylene)-2-(4-(tert-butyl)phenyl)oxazol-5(4H)-one (F11)

Chem.

[0182] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 9.23 (s, 1H), 8.31 (d, 1H, J = 8.0 Hz), 8.09 (d, 3H, J = 8.4 Hz), 7.64 - 7.62 (m, 3H), 7.53 - 7.44 (m, 2H), 1.31 (s, 9H) ESI (m / z) 362 (MH + )

[0183] Example 1.12: 4-((1H-Pyrrol-2-yl)methylene)-2-(4-isopropylphenyl)oxazol-5(4H)-one (F12)

Chem.

[0184] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 11.52 (s, 1H), 8.08 (d, 2H, J = 8.0 Hz), 7.43 (d, 2H, J = 8.0 Hz), 7.32 (s, 1H), 7.19 (s, 1H), 7.05 (s, 1H), 6.34 (s, 1H), 3.01 - 2.91 (m, 1H), 1.21 (d, 6H, J = 6.8 Hz)

[0185] ESI (m / z) 281 (MH + )

[0186] Example 1.13: 2-(4-(tert-butyl)phenyl)-4-(quinolin-4-ylmethylene)oxazol-5(4H)-one (F13) [Chemical]

[0187] 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.08 (d, 1H, J = 4.4 Hz), 8.64 (d, 1H, J = 4.8 Hz), 8.44 (d, 1H, J = 8.4 Hz), 8.10 - 8.07 (m, 3H), 7.95 (s, 1H), 7.83 (t, 1H, J = 7.6 Hz), 7.71 (d, 1H, J = 8.0 Hz), 7.66 (d, 2H, J = 8.0 Hz), 1.31 (s, 9H)

[0188] ESI (m / z) 357 (MH + )

[0189] Example 1.14: 2-(4-Isobutylphenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F14) [Chemical]

[0190] 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.01 (d, 1H, J = 5.2 Hz), 7.96 (d, 2H, J = 7.2 Hz), 7.81 (d, 1H, J = 3.2 Hz), 7.67 (s, 1H), 7.39 (d, 2H, J = 7.2 Hz), 7.22 (t, 1H, J = 4.4 Hz), 2.53 (d, 2H, J = 6.8 Hz), 1.91 - 1.81 (m, 1H), 0.84 (d, 6H, J = 6.4 Hz)

[0191] ESI (m / z) 312 (MH + )

[0192] Example 1.15: 2-(4-Isobutylphenyl)-4-(thiophen-3-ylmethylene)oxazol-5(4H)-one (F15) [Chemical]

[0193] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.45 (d, 1H, J = 2.4 Hz), 8.01 - 7.99 (m, 3H), 7.69 (q, 1H, J = 2.8 Hz), 7.39 (s, 1H), 7.37 (s, 2H), 2.53 (d, 2H, J = 7.2 Hz), 1.92 - 1.81 (m, 1H), 0.85 (d, 6H, J = 6.8 Hz)

[0194] ESI (m / z) 312 (MH + )

[0195] Example 1.16: 4-(Benzo[b]thiophen-3-ylmethylene)-2-(4-isopropylphenyl)oxazol-5(4H)-one (F16)

Chemical Structure

[0196] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 9.24 (s, 1H), 8.30 (d, 1H, J = 7.6 Hz), 8.09 (m, 3H), 7.61 (s, 1H), 7.52 - 7.44 (m, 4H), 3.03 - 2.96 (m, 1H), 1.22 (d, 6H, J = 6.8 Hz)

[0197] Example 1.17: 4-((1H-Pyrrol-2-yl)methylene)-2-(4-(trifluoromethyl)phenyl)oxazol-5(4H)-one (F17)

Chemical Structure

[0198] 1 1H NMR (400 MHz, DMSO-d 6)δ 11.67 (s, 1H), 8.39 (d, 2H, J = 7.6 Hz), 7.95 (d, 2H, J = 8.0 Hz), 7.42 (s, 1H), 7.33 (s, 1H), 7.15 (s, 1H), 6.41 (s, 1H)

[0199] ESI (m / z) 305 (MH-), 307 (MH + )

[0200] Example 1.18: 4-(Thiophen-3-ylmethylene)-2-(4-(trifluoromethyl)phenyl)oxazol-5(4H)-one (F18)

Chemical Structure

[0201] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.55 (d, 1H, J = 1.9 Hz), 8.32 (d, 2H, J = 8.1 Hz), 8.07 (d, 1H, J = 5.0 Hz), 7.98 (d, 2H, J = 8.2 Hz), 7.78 - 7.74 (m, 1H), 7.53 (s, 1H)

[0202] Example 1.19: 4-(Furan-2-ylmethylene)-2-(4-isobutylphenyl)oxazol-5(4H)-one (F19)

Chemical Structure

[0203] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.04 (s, 1H), 7.97 (d, 2H, J = 8.0 Hz), 7.55 (d, 1H, J = 3.6 Hz), 7.36 (d, 2H, J = 8.0 Hz), 7.14 (s, 1H), 6.79 (d, 1H, J = 2.0 Hz), 2.52 (d, 2H, J = 6.4 Hz), 1.90 - 1.80 (m, 1H), 0.84 (d, 6H, J = 6.4 Hz)

[0204] Example 1.20: 4-((1H-Pyrrol-2-yl)methylene)-2-(4-isobutylphenyl)oxazol-5(4H)-one (F20)

Chemical formula

[0205] 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.51 (s, 1H), 8.06 (d, 2H, J = 8.0 Hz), 7.34 (d, 2H, J = 8.4 Hz), 7.31 (s, 1H), 7.18 (s, 1H), 7.04 (s, 1H), 6.33 (s, 1H), 2.51 (d, 2H, J = 6.8 Hz), 1.90 - 1.80 (m, 1H), 0.84 (d, 6H, J = 6.4 Hz)

[0206] Example 1.21: 4-(Benzo[b]thiophen-3-ylmethylene)-2-(4-isobutylphenyl)oxazol-5(4H)-one (F21)

Chemical formula

[0207] 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.24 (s, 1H), 8.29 (d, 1H, J = 8.0 Hz), 8.08 (d, 3H, J = 8.0 Hz), 7.60 (s, 1H), 7.52 - 7.43 (m, 2H), 7.39 (d, 2H, J = 7.6 Hz), 2.54 (d, 2H, J = 6.8 Hz), 1.89 - 1.84 (m, 1H), 0.85 (d, 6H, J = 6.4 Hz)

[0208] Example 1.22: 2-(4-Bromophenyl)-4-((4-methylthiazol-5-yl)methylene)oxazol-5(4H)-one (F22)

Chemical formula

[0209] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 9.31 (s, 1H), 7.96 (d, 2H, J = 8.4 Hz), 7.83 (d, 2H, J = 8.4 Hz), 7.63 (s, 1H), 2.62 (s, 3H)

[0210] ESI (m / z) 350 (MH + )

[0211] Example 1.23: 2-Cyclohexyl-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F23)

Chem.

[0212] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 7.96 (d, 1H, J = 5.2 Hz), 7.74 (d, 1H, J = 3.6 Hz), 7.57 (s, 1H), 7.18 (t, 1H, J = 4.2 Hz), 2.71 - 2.66 (m, 1H), 1.96 - 1.93 (m, 2H), 1.74 - 1.71 (m, 2H), 1.61 - 1.58 (m, 1H), 1.53 - 1.44 (m, 2H), 1.38 - 1.18 (m, 3H)

[0213] ESI (m / z) 262 (MH + )

[0214] Example 1.24: 2-([1,1'-Biphenyl]-4-yl)-4-((4-methylthiazol-5-yl)methylene)oxazol-5(4H)-one (F24)

Chem.

[0215] 1 1H NMR (400 MHz, DMSO-d 6)δ 9.31 (s, 1H), 8.12 (d, 2H, J = 8.4 Hz), 7.93 (d, 2H, J = 8.0 Hz), 7.77 (d, 2H, J = 7.6 Hz), 7.61 (s, 1H), 7.50 (t, 2H, J = 7.4 Hz), 7.43 (t, 1H, J = 7.4 Hz), 2.63 (s, 3H)

[0216] Example 1.25: 4 - ((1H - Pyrrol - 2 - yl)methylene)-2 - ([1,1’ - biphenyl]-4 - ylmethyl)oxazol - 5(4H)-one (F25)

Chem.

[0217] 1 H NMR (400 MHz, MHz, DMSO - d 6 )δ 11.44 (s, 1H), 7.63 (d, 4H, J = 8.0 Hz), 7.44 - 7.41 (m, 4H), 7.34 - 7.31 (m, 1H), 7.25 (s, 1H), 7.13 (s, 1H), 7.02 (s, 1H), 6.30 (s, 1H), 4.05 (s, 2H)

[0218] ESI (m / z) 329 (MH + )、327 (MH - )

[0219] Example 1.26: 4 - ((1H - Pyrrol - 2 - yl)methylene)-2 - (4 - butylphenyl)oxazol - 5(4H)-one (F26)

Chem.

[0220] 1 H NMR (400 MHz, MHz, DMSO - d 6) δ 11.56 (s, 1H), 8.11 (d, 2H, J = 7.9 Hz), 7.43 (d, 2H, J = 8.0 Hz), 7.35 (s, 1H), 7.22 (s, 1H), 7.08 (s, 1H), 6.37 (s, 1H), 2.69 (t, 2H, J = 7.6 Hz), 1.65 - 1.55 (m, 2H), 1.38 - 1.27 (m, 2H), 0.91 (t, 3H, J = 7.3 Hz)

[0221] Example 1.27: 2-(4-(tert-Butyl)phenyl)-4-((1-phenyl-1H-pyrrol-2-yl)methylene)oxazol-5(4H)-one (F27)

Chemical Structure

[0222] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 7.98 (d, 2H, J = 8.4 Hz), 7.77 (d, 1H, J = 3.9 Hz), 7.62 - 7.57 (m, 4H), 7.55 - 7.51 (m, 1H), 7.50 - 7.48 (m, 1H), 7.45 (d, 2H, J = 7.5 Hz), 6.74 (s, 1H), 6.58 (t, 1H, J = 3.4 Hz), 1.30 (s, 9H)

[0223] Example 1.28: 2-([1,1'-Biphenyl]-4-ylmethyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F28)

Chemical Structure

[0224] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 7.98 (d, 1H, J = 4.8 Hz), 7.76 (d, 1H, J = 3.6 Hz), 7.65 (s, 2H), 7.63 (s, 3H), 7.46 - 7.41 (m, 4H), 7.33 (t, 1H, J = 7.2 Hz), 7.19 (t, 1H, J = 4.4 Hz), 4.10 (s, 2H)

[0225] Example 1.29: 2-(2,3-Dihydro-1H-inden-5-yl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F29)

Chem.

[0226] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.01 (d, 1H, J = 5.2 Hz), 7.89 (s, 1H), 7.83 (d, 1H, J = 7.6 Hz), 7.80 (d, 1H, J = 3.6 Hz), 7.65 (s, 1H), 7.43 (d, 1H, J = 8.0 Hz), 7.23 - 7.21 (m, 1H), 2.93 (t, 4H, J = 7.3 Hz), 2.08 - 2.00 (m, 2H)

[0227] ESI (m / z) 296 (MH + )

[0228] Example 1.30: 4-((1H-Pyrrol-2-yl)methylene)-2-(naphthalen-1-yl)oxazol-5(4H)-one (F30)

Chem.

[0229] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 11.62 (s, 1H), 9.31 (d, 1H, J = 8.0 Hz), 8.29 (d, 1H, J = 7.2 Hz), 8.20 (d, 1H, J = 8.4 Hz), 8.06 (d, 1H, J = 8.0 Hz), 7.76 (t, 1H, J = 7.6 Hz), 7.65 (q, 2H, J = 8.4 Hz), 7.36 (s, 1H), 7.28 (s, 2H), 6.43 (t, 1H, J = 2.8 Hz)

[0230] ESI (m / z) 289 (MH + )

[0231] Example 1.31: Methyl 4-(4-(furan-2-ylmethylene)-5-oxo-4,5-dihydrooxazol-2-yl)benzoate (F31)

Chem.

[0232] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.21 (s, 1H), 8.18 (s, 1H), 8.13 (s, 1H), 8.11 - 8.10 (m, 2H), 7.60 (d, 1H, J = 3.2 Hz), 7.27 (s, 1H), 6.84 - 6.82 (m, 1H), 3.87 (s, 3H)

[0233] Example 1.32: Methyl 4-(5-oxo-4-(thiophen-3-ylmethylene)-4,5-dihydrooxazol-2-yl)benzoate (F32)

Chem.

[0234] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.47 (s, 1H), 8.16 - 8.00 (m, 5H), 7.70 (s, 1H), 7.44 (s, 1H), 3.85 (s, 3H)

[0235] Example 1.33: N-(4-(5-oxo-4-(thiophen-2-ylmethylene)-4,5-dihydrooxazol-2-yl)phenyl)acetamide (F33)

Chem.

[0236] Example 1.34: 2-(2-Methoxyphenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F34)

Chem.

[0237] Example 1.35: N-(tert-Butyl)-4-(5-oxo-4-(thiophen-2-ylmethylene)-4,5-dihydrooxazol-2-yl)benzenesulfonamide (F35)

Chem.

[0238] Example 1.36: N-Isopropyl-4-(5-oxo-4-(thiophen-2-ylmethylene)-4,5-dihydrooxazol-2-yl)benzenesulfonamide (F36)

Chem.

[0239] Example 1.37: 2-([1,1'-Biphenyl]-4-yl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F37)

Chem.

[0240] 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.23 (d, 2H, J = 8.3 Hz), 7.79 - 7.71 (m, 3H), 7.69 - 7.61 (m, 3H), 7.53 - 7.46 (m, 3H), 7.45 - 7.38 (m, 1H), 7.17 (t, 1H)

[0241] ESI (m / z) 332 (MH + )

[0242] Example 1.38: 2-(4-(tert-Butyl)phenyl)-4-(furan-2-ylmethylene)oxazol-5(4H)-one (F38)

Chem.

[0243] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.05 (s, 1H), 7.99 (d, 2H, J = 8.4 Hz), 7.60 (d, 2H, J = 8.4 Hz), 7.55 (d, 1H, J = 3.2 Hz), 7.15 (s, 1H), 6.81 (br s, 1H), 1.29 (s, 9H)

[0244] ESI (m / z) 296 (MH + )

[0245] Example 1.39: 2-(4-(tert-Butyl)phenyl)-4-((1-methyl-1H-pyrazol-4-yl)methylene)oxazol-5(4H)-one (F39)

Chem.

[0246] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.52 (s, 1H), 8.18 (s, 1H), 8.05 (d, 2H, J = 8.5 Hz), 7.63 (d, 2H, J = 8.5 Hz), 7.35 (s, 1H), 3.96 (s, 3H), 1.34 (s, 9H)

[0247] ESI (m / z) 310 (MH + )

[0248] Example 1.40: 2-(4-(tert-Butyl)phenyl)-4-(pyridin-3-ylmethylene)oxazol-5(4H)-one (F40)

Chem.

[0249] 1 1H NMR (400 MHz, DMSO-d 6)δ 9.28 (d, 1H, J = 1.9 Hz), 8.78 (dt, 1H, J = 8.1, 1.7 Hz), 8.65 (dd, 1H, J = 4.8, 1.6 Hz), 8.09 (d, 2H), 7.68 (d, 2H, J = 8.6 Hz), 7.61 - 7.56 (m, 1H), 7.40 (s, 1H), 1.34 (s, 9H)

[0250] ESI(m / z) 307 (MH + )

[0251] Example 1.41: 2-(4-(tert-Butyl)phenyl)-4-(pyridin-4-ylmethylene)oxazol-5(4H)-one (F41)

Chemical Structure

[0252] 1 H NMR (400 MHz, MHz, DMSO-d 6 )δ 8.71 (d, 2H, J = 5.2 Hz), 8.13 (d, 2H, J = 5.5 Hz), 8.07 (d, 2H), 7.66 (d, 2H, J = 8.3 Hz), 7.27 (s, 1H), 1.31 (s, 9H)

[0253] ESI(m / z) 307 (MH + )

[0254] Example 1.42: 2-([1,1'-Biphenyl]-4-yl)-4-(pyridin-3-ylmethylene)oxazol-5(4H)-one (F42)

Chemical Structure

[0255] 1 H NMR (400 MHz, MHz, DMSO-d 6)δ 9.25 (s, 1H), 8.78 (d, 1H, J = 7.8 Hz), 8.63 (s, 1H), 8.19 (d, 2H, J = 7.8 Hz), 7.92 (d, 2H, J = 7.9 Hz), 7.77 (d, 2H, J = 7.1 Hz), 7.61 - 7.54 (m, 1H), 7.50 (t, 2H, J = 6.9 Hz), 7.47 - 7.36 (m, 2H)

[0256] ESI (m / z) 327 (MH + )

[0257] Example 1.43: 2-(4-(tert-Butyl)phenyl)-4-(4-(methylthio)benzylidene)oxazol-5(4H)-one (F43)

Chemical Structure

[0258] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.20 (br s, 2H), 8.00 (br s, 2H), 7.63 (br s, 2H), 7.37 (s, 2H), 7.27 (s, 1H), 2.53 (s, 3H), 1.31 (s, 9H)

[0259] ESI (m / z) 352 (MH + )

[0260] Example 1.44: 4-Benzylidene-2-(4-(tert-butyl)phenyl)oxazol-5(4H)-one (F44)

Chemical Structure

[0261] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.26 (d, 2H, J = 6.8 Hz), 8.00 (d, 2H, J = 8.4 Hz), 7.61 (d, 2H, J = 8.0 Hz), 7.52 - 7.45 (m, 3H), 7.28 (s, 1H), 1.29 (s, 9H)

[0262] ESI(m / z) 306 (MH + )

[0263] Example 1.45: 2-(4-Bromophenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F45)

Chemical Structure

[0264] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.05 (s, 1H), 7.95 (d, 2H, J = 7.6 Hz), 7.83 (s, 2H), 7.81 (s, 1H), 7.73 (s, 1H), 7.23 (s, 1H)

[0265] Example 1.46: 2-(4-Isopropylphenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F46)

Chemical Structure

[0266] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.02 (d, 1H, J = 5.2 Hz), 7.97 (d, 2H, J = 8.0 Hz), 7.81 (d, 1H, J = 3.6 Hz), 7.67 (s, 1H), 7.48 (d, 2H, J = 8.4 Hz), 7.22 (t, 1H, J = 4.4 Hz), 3.02 - 2.92 (m, 1H), 1.21 (d, 6H, J = 6.8 Hz)

[0267] ESI(m / z) 298 (MH + )

[0268] Example 1.47: 2-(4-Isopropylphenyl)-4-(thiophen-3-ylmethylene)oxazol-5(4H)-one (F47)

Chemical Structure

[0269] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.45 (d, 1H, J = 2.8 Hz), 8.02 - 8.00 (m, 3H), 7.70 (q, 1H, J = 2.8 Hz), 7.47 (d, 2H, J = 7.2 Hz), 7.38 (s, 1H), 3.03 - 2.93 (m, 1H), 1.21 (d, 6H, J = 6.8 Hz)

[0270] ESI (m / z) 298 (MH + )

[0271] Example 1.48: 4-(Furan-2-ylmethylene)-2-(4-(trifluoromethyl)phenyl)oxazol-5(4H)-one (F48)

Chem.

[0272] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.25 (d, 2H, J = 8.2 Hz), 8.10 (s, 1H), 7.94 (d, 2H, J = 8.3 Hz), 7.60 (d, 1H, J = 3.5 Hz), 7.28 (s, 1H), 6.83 - 6.81 (m, 1H)

[0273] Example 1.49: 4-(Furan-2-ylmethylene)-2-(4-isopropylphenyl)oxazol-5(4H)-one (F49)

Chem.

[0274] 1 H NMR (400 MHz, MHz, DMSO-d 6) δ 8.04 (s, 1H), 7.97 (d, 2H, J = 8.1 Hz), 7.54 (d, 1H, J = 3.3 Hz), 7.44 (d, 2H, J = 8.1 Hz), 7.13 (s, 1H), 6.83 - 6.76 (m, 1H), 3.01 - 2.88 (m, 1H), 1.19 (d, 6H, J = 6.9 Hz)

[0275] Example 1.50: 2-(4-Bromophenyl)-4-(furan-2-ylmethylene)oxazol-5(4H)-one (F50)

Chemical Structure

[0276] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.11 (s, 1H), 8.07 - 7.98 (m, 2H), 7.88 - 7.77 (m, 2H), 7.62 (s, 1H), 7.25 (s, 1H), 6.85 (s, 1H)

[0277] Example 1.51: 2-(4-Bromophenyl)-4-(thiophen-3-ylmethylene)oxazol-5(4H)-one (F51)

Chemical Structure

[0278] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.47 (d, 1H, J = 1.6 Hz), 8.01 - 7.99 (m, 3H), 7.80 (s, 1H), 7.78 (s, 1H), 7.70 - 7.68 (m, 1H), 7.43 (s, 1H)

[0279] Example 1.52: 2-(Naphthalen-2-yl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F52)

Chemical Structure

[0280] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.70 (s, 1H), 8.52 (d, 1H, J = 2.4 Hz), 8.21 - 8.17 (m, 2H), 8.12 (s, 1H), 8.09 (t, 1H, J = 4.8 Hz), 8.02 (d, 1H, J = 8.0 Hz), 7.74 - 7.72 (m, 1H), 7.69 - 7.61 (m, 2H), 7.44 (s, 1H)

[0281] ESI (m / z) 306 (MH + )

[0282] Example 1.53: 2-(4-Butylphenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F53) [Chemical Structure]

[0283] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.02 (d, 1H, J = 5.0 Hz), 7.96 (d, 2H, J = 8.2 Hz), 7.81 (d, 1H, J = 3.6 Hz), 7.67 (s, 1H), 7.42 (d, 2H, J = 8.2 Hz), 7.25 - 7.20 (m, 1H), 2.66 (t, 2H, J = 7.7 Hz), 1.61 - 1.51 (m, 2H), 1.37 - 1.22 (m, 2H), 0.87 (t, 3H, J = 7.3 Hz)

[0284] Example 1.54: 2-(4-Butylphenyl)-4-(furan-2-ylmethylene)oxazol-5(4H)-one (F54) [Chemical Structure]

[0285] 1 1H NMR (400 MHz, DMSO-d 6) δ 8.09 (d, 1H, J = 1.5 Hz), 8.02 (d, 2H, J = 8.2 Hz), 7.60 (d, 1H, J = 3.5 Hz), 7.45 (d, 2H, J = 8.2 Hz), 7.19 (s, 1H), 6.86 - 6.83 (m, 1H), 2.69 (t, 2H, J = 7.7 Hz), 1.66 - 1.54 (m, 2H), 1.39 - 1.27 (m, 2H), 0.91 (t, 3H, J = 7.3 Hz)

[0286] Example 1.55: 2-(4-(Difluoromethoxy)phenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F55)

Chemical Structure

[0287] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.10 (d, 2H, J = 8.8 Hz), 8.03 (d, 1H, J = 5.2 Hz), 7.82 (d, 1H, J = 3.6 Hz), 7.69 (s, 1H), 7.41 (s, 1H), 7.38 (d, 2H, J = 8.8 Hz), 7.24 - 7.22 (m, 1H)

[0288] Example 1.56: 2-(4-(tert-Butyl)phenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F56)

Chemical Structure

[0289] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.02 (d, 1H, J = 5.2 Hz), 7.96 (d, 2H, J = 7.6 Hz), 7.80 (d, 1H, J = 3.6 Hz), 7.66 (s, 1H), 7.61 (d, 2H, J = 7.6 Hz), 7.22 (t, 1H, J = 4.0 Hz), 1.30 (s, 9H)

[0290] 1313C NMR (400 MHz, DMSO-d 6 ) δ 166.7, 162.1, 157.2, 137.6, 136.9, 136.6, 130.7, 129.6, 129.1, 129.1, 126.4, 126.4, 124.9, 122.8, 35.46, 31.2, 31.2, 31.2

[0291] Example 1.57: 4-(2-Nitrobenzylidene)-2-phenyloxazol-5(4H)-one (F57)

Chem.

[0292] Example 1.58: 4-((2-(4-(tert-Butyl)phenyl)-5-oxooxazol-4(5H)-ylidene)methyl)phenyl acetate (F58)

Chem.

[0293] Example 1.59: 2-(3-Chlorobenzo[b]thiophen-2-yl)-4-(4-(dimethylamino)benzylidene)oxazol-5(4H)-one (F59)

Chem.

[0294] Example 1.60: 4-((2-4-Methoxyphenyl)-5-oxooxazol-4(5H)-ylidene)methyl)benzoic acid (F60)

Chem.

[0295] Example 1.61: 2-Phenyl-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F61)

Chem.

[0296] Example 1.62: 4-(4-Isopropylbenzylidene)-2-(naphthalen-1-yl)oxazol-5(4H)-one (F62)

Chem.

[0297] Example 1.63: 2-(3-Iodo-4-methylphenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F63)

Chem.

[0298] Example 1.64: 4-(4-(Benzyloxy)benzylidene)-2-(4-(tert-butyl)phenyl)oxazol-5(4H)-one (F64)

Chem.

[0299] Example 1.65: 4-((1-(2-Chlorobenzyl)-1H-pyrrol-2-yl)methylene)-2-phenyloxazol-5(4H)-one (F65)

Chem.

[0300] Example 1.66: N-(4-(4-(2-(Allyloxy)-4-(diethylamino)benzylidene)-5-oxo-4,5-dihydrooxazol-2-yl)phenyl)-5-bromonicotinamide (F66)

Chem.

[0301] Example 1.67: 4-((2-(2-Chlorophenyl)-5-oxooxazol-4(5H)-ylidene)methyl)phenyl 4-(tert-butyl)benzoate (F67)

Chem.

[0302] Example 1.68: 4-((Z)-1-(3-Ethyl-5-methoxybenzo[d]thiazol-2(3H)-ylidene)butane-2-ylidene)-2-phenyloxazol-5(4H)-one (F68)

Chem.

[0303] Example 1.69: 4-((2-(4-Acetamidophenyl)-5-oxooxazol-4(5H)-ylidene)methyl)-2-ethoxyphenyl thiophene-2-carboxylate (F69)

Chem.

[0304] Example 1.70: 4-((6-Ethoxy-2-(phenylthio)quinolin-3-yl)methylene)-2-phenyloxazol-5(4H)-one (F70)

Chem.

[0305] Example 1.71: 2-(3-Bromophenyl)-4-((5-chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)methylene)oxazol-5(4H)-one (F71)

Chem.

[0306] Example 1.72: 4-((1-Acetyl-1H-indol-3-yl)methylene)-2-(4-bromophenyl)oxazol-5(4H)-one (F72)

Chem.

[0307] Example 1.73: 2-(4-(tert-Butyl)phenyl)-4-((5-(3-(trifluoromethyl)phenyl)furan-2-yl)methylene)oxazol-5(4H)-one (F73)

Chem.

[0308] Example 1.74: 4-((7-Methoxy-2-oxo-1,2-dihydroquinolin-3-yl)methylene)-2-phenyloxazol-5(4H)-one (F74)

Chem.

[0309] Example 1.75: 2-(4-Methoxyphenyl)-4-(2-(thiophen-2-yl)-4H-chromen-4-ylidene)oxazol-5(4H)-one (F75)

Chem.

[0310] Example 1.76: N,N-Dimethyl-3-((5-oxo-2-(p-tolyl)oxazol-4(5H)-ylidene)methyl)-1H-indole-1-sulfonamide (F76)

Chem.

[0311] Example 1.77: 2-((2-(4-Chlorophenyl)-5-oxooxazol-4(5H)-ylidene)methyl)phenyl 4-acetamidobenzenesulfonate (F77)

Chem.

[0312] Example 1.78: 4-((5-Oxo-2-phenyloxazol-4(5H)-ylidene)methyl)phenyl furan-2-carboxylate (F78)

Chem.

[0313] Example 1.79: 2,2'-(1,4-Phenylene)bis(4-((5-methylfuran-2-yl)methylene)oxazol-5(4H)-one) (F79)

Chem.

[0314] Example 1.80: 2-((2-(4-(tert-Butyl)phenyl)-5-oxooxazol-4(5H)-ylidene)methyl)phenyl benzenesulfonate (F80)

Chem.

[0315] Example 1.81: 4-((1-Acetyl-1H-indol-3-yl)methylene)-2-(4-(tert-butyl)phenyl)oxazol-5(4H)-one (F81)

Chem.

[0316] Example 1.82: 4-((1-Acetyl-3-phenyl-1H-pyrazol-4-yl)methylene)-2-(4-(tert-butyl)phenyl)oxazol-5(4H)-one (F82)

Chem.

[0317] Example 1.83: 4-((1,3-Diphenyl-1H-pyrazol-4-yl)methylene)-2-(p-tolyl)oxazol-5(4H)-one (F83)

Chem.

[0318] Example 1.84: 4-((6-Methoxy-2-oxo-1,2-dihydroquinolin-3-yl)methylene)-2-phenyloxazol-5(4H)-one (F84)

Chem.

[0319] Example 1.85: 4-(2,6-Diphenyl-4H-thiopyran-4-ylidene)-2-phenyloxazol-5(4H)-one (F85)

Chem.

[0320] Example 1.86: N-(4-(4-(4-((2-Chloroethyl)(methyl)amino)benzylidene)-5-oxo-4,5-dihydrooxazol-2-yl)phenyl)acetamide (F86)

Chem.

[0321] Example 1.87: 2-(4-Methoxyphenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F87)

Chem.

[0322] Example 1.88: 2-(4-Chlorophenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F88)

Chem.

[0323] Example 1.89: 2-(Thiophen-2-yl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F89)

Chem.

[0324] Example 1.90: 4-(5-Oxo-4-(thiophen-2-ylmethylene)-4,5-dihydrooxazol-2-yl)phenyl acetate (F90)

Chem.

[0325] Example 1.91: 2-(4-Chloro-3-nitrophenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F91)

Chem.

[0326] Example 1.92: 2-(2-Chlorophenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F92)

Chem.

[0327] Example 1.93: 2-(2-Chloro-4-nitrophenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F93)

Chem.

[0328] Example 1.94: 4-(Thiophen-2-ylmethylene)-2-(3,4,5-trimethoxyphenyl)oxazol-5(4H)-one (F94)

Chemical Structure

[0329] Example 1.95: 2-(4-Phenoxyphenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F95)

Chemical Structure

[0330] Example 1.96: 2-(4-(tert-Butyl)phenyl)-4-((5-methylthiophen-2-yl)methylene)oxazol-5(4H)-one (F96)

Chemical Structure

[0331] Example 1.97: 2-(Benzod][1,3]dioxol-5-yl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F97)

Chemical Structure

[0332] Example 1.98: 2-(4-(2-Oxopyrrolidin-1-yl)phenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F98)

Chemical Structure

[0333] Example 1.99: 4-((5-Chlorothiophen-2-yl)methylene)-2-(4-methoxyphenyl)oxazol-5(4H)-one (F99) [Chemical formula]

[0334] Example 1.100: 2-(Benzod[d][1,3]dioxol-5-yl)-4-((5-(piperidin-1-yl)thiophen-2-yl)methylene)oxazol-5(4H)-one (F100) [Chemical formula]

[0335] Example 1.101: 2-(3,4-Diethoxyphenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F101) [Chemical formula]

[0336] Example 1.102: 2-(4-(Difluoromethoxy)-3-methoxyphenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F102) [Chemical formula]

[0337] Example 1.103: 2-(5-Ethyl-4-methylthiophen-2-yl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F103) [Chemical formula]

[0338] Example 1.104: 2-(3-Methoxyphenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F104) [Chemical formula]

[0339] Example 1.105: 4-((5-(Dimethylamino)thiophen-2-yl)methylene)-2-(naphthalen-2-yl)oxazol-5(4H)-one (F105)

Chem.

[0340] Example 1.106: N-(4-(4-((4-Bromothiophen-2-yl)methylene)-5-oxo-4,5-dihydrooxazol-2-yl)phenyl)acetamide (F106)

Chem.

[0341] Example 1.107: N-(4-(4-((5-Bromothiophen-2-yl)methylene)-5-oxo-4,5-dihydrooxazol-2-yl)phenyl)acetamide (F107)

Chem.

[0342] Example 1.108: N-(4-(4-((5-(Dimethylamino)thiophen-2-yl)methylene)-5-oxo-4,5-dihydrooxazol-2-yl)phenyl)acetamide (F108)

Chem.

[0343] Example 1.109: 2-(4-(Benzyloxy)phenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F109)

Chem.

[0344] Example 1.110: 2-((E)-Styryl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F110)

Chem.

[0345] Example 1.111: 2-(5-Methylthiophen-2-yl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F111)

Chem.

[0346] Example 1.112: N,N-Diethyl-3-(5-oxo-4-(thiophen-2-ylmethylene)-4,5-dihydrooxazol-2-yl)benzenesulfonamide (F112)

Chem.

[0347] Example 1.113: 2-(2-Methoxyphenyl)-4-((2,4,5-trimethylthieno[2,3-d]pyrimidin-6-yl)methylene)oxazol-5(4H)-one (F113)

Chem.

[0348] Example 1.114: 4-((5-(Dimethylamino)thiophen-2-yl)methylene)-2-(2-methoxyphenyl)oxazol-5(4H)-one (F114)

Chem.

[0349] Example 1.115: 4-((5-Methylthiophen-2-yl)methylene)-2-(thiophen-2-yl)oxazol-5(4H)-one (F115)

Chem.

[0350] Example 1.116: N,N-Dimethyl-3-(5-oxo-4-(thiophen-2-ylmethylene)-4,5-dihydrooxazol-2-yl)benzenesulfonamide (F116)

Chem.

[0351] Example 1.117: 4-((5-Bromothiophen-2-yl)methylene)-2-(2-methoxyphenyl)oxazol-5(4H)-one (F117)

Chem.

[0352] Example 1.118: 4-((5-Bromothiophen-2-yl)methylene)-2-(thiophen-2-yl)oxazol-5(4H)-one (F118)

Chem.

[0353] Example 1.119: 2-(5-Methylthiophen-2-yl)-4-((5-methylthiophen-2-yl)methylene)oxazol-5(4H)-one (F119)

Chem.

[0354] Example 1.120: 4-((5-(Piperidin-1-yl)thiophen-2-yl)methylene)-2-(thiophen-2-yl)oxazol-5(4H)-one (F120)

Chem.

[0355] Example 1.121: 4-((5-Bromothiophen-2-yl)methylene)-2-(2-(difluoromethoxy)phenyl)oxazol-5(4H)-one (F121)

Chem.

[0356] Example 1.122: 2-(4-(tert-Butyl)phenyl)-4-(2-(difluoromethoxy)benzylidene)oxazol-5(4H)-one (F122)

Chem.

[0357] Example 1.123: 4-((1-(2-Chlorobenzyl)-1H-pyrazol-4-yl)methylene)-2-(naphthalen-2-yl)oxazol-5(4H)-one (F123)

Chem.

[0358] Example 1.124: 2-(4-(tert-Butyl)phenyl)-4-((1-phenyl-1H-pyrazol-4-yl)methylene)oxazol-5(4H)-one (F124)

Chem.

[0359] Example 1.125: 2-(2-Methoxyphenyl)-4-((1-methyl-1H-pyrazol-4-yl)methylene)oxazol-5(4H)-one (F125)

Chem.

[0360] Example 1.126: N-(tert-Butyl)-4-(4-((5-methylfuran-2-yl)methylene)-5-oxo-4,5-dihydrooxazol-2-yl)benzenesulfonamide (F126)

Chem.

[0361] Example 1.127: N,N-Diethyl-4-(4-(1-methylpyridin-2(1H)-ylidene)-5-oxo-4,5-dihydrooxazol-2-yl)benzenesulfonamide (F127)

Chem.

[0362] Example 1.128: 2-(4-((2-(2-Methoxyphenyl)-5-oxooxazol-4(5H)-ylidene)methyl)phenoxy)acetamide (F128)

Chem.

[0363] Example 1.129: N-(4-((2-(4-Isopropoxyphenyl)-5-oxooxazol-4(5H)-ylidene)methyl)phenyl)acetamide (F129)

Chem.

[0364] Example 1.130: 4-((1-Benzyl-1H-pyrazol-4-yl)methylene)-2-(4-isopropoxyphenyl)oxazol-5(4H)-one (F130)

Chem.

[0365] Example 1.131: N,N-Diethyl-3-(4-((1-methyl-1H-pyrazol-4-yl)methylene)-5-oxo-4,5-dihydrooxazol-2-yl)benzenesulfonamide (F131)

Chem.

[0366] Example 1.132: 4-(4-(1H-1,2,4-triazol-1-yl)benzylidene)-2-(2-iodophenyl)oxazol-5(4H)-one (F132)

Chem.

[0367] Example 1.133: 4-(4-((3,5-dimethylisoxazol-4-yl)methoxy)-3-methoxybenzylidene)-2-(2-methoxyphenyl)oxazol-5(4H)-one (F133)

Chem.

[0368] Example 1.134: N-(4-(4-(4-(1H-1,2,4-triazol-1-yl)benzylidene)-5-oxo-4,5-dihydrooxazol-2-yl)phenyl)acetamide (F134)

Chem.

[0369] Example 1.135: 2-(2-(Difluoromethoxy)phenyl)-4-((E)-3-(furan-2-yl)allylidene)oxazol-5(4H)-one (F135)

Chem.

[0370] Example 1.136: 4-((1-(tert-Butyl)-1H-pyrazol-4-yl)methylene)-2-(2-methoxyphenyl)oxazol-5(4H)-one (F136)

Chem.

[0371] Example 1.137: 4-((4-Methylthiazol-5-yl)methylene)-2-(4-propylphenyl)oxazol-5(4H)-one (F137)

Chem.

[0372] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 9.27 (s, 1H), 9.94 (d, 2H, J = 8.0 Hz), 7.53 (s, 1H), 7.41 (d, 2H, J = 8.0 Hz), 2.63 (t, 2H, J = 7.6 Hz), 2.59 (s, 3H), 1.64 - 1.55 (m, 2H), 0.87 (t, 3H, J = 7.2 Hz) 13 C NMR (400 MHz, DMSO-d 6 ) δ 166.1, 12.6, 160.62, 159.76, 149.2, 132.0, 129.9, 129.3, 126.5, 123.0, 121.3, 37.7, 24.1, 16.2, 14.0

[0373] Example 1.138: 4-((1H-Pyrrol-2-yl)methylene)-2-(4-butoxyphenyl)oxazol-5(4H)-one (F138)

Chem.

[0374] 1 H NMR (400 MHz, MHz, DMSO-d 6) δ 11.50 (s, 1H), 8.08 (d, 2H, J = 8.4 Hz), 7.29 (s, 1H), 7.13 (s, 1H), 7.07 (d, 2H, J = 8.4 Hz), 7.00 (s, 1H), 6.31 (s, 1H), 4.03 (t, 2H, J = 6.0 Hz), 1.70 - 1.67 (m, 2H), 1.45 - 1.36 (m, 2H), 0.91 (t, 3H, J = 5.2 Hz)

[0375] Example 1.139: 2-(4-Hexylphenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F139) [Chemical formula]

[0376] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.01 (d, 1H, J = 3.6 Hz), 7.95 (d, 2H, J = 8.0 Hz), 7.80 (br s, 1H), 7.66 (s, 1H), 7.41 (d, 2H, J = 8.0 Hz), 7.22 (br s, 1H), 2.64 (t, 2H, J = 7.2 Hz), 1.56 (brs 2H), 1.24 (s, 6H), 0.81 (br s, 3H)

[0377] 13 C NMR (400 MHz, DMSO-d 6 ) δ 166.7, 162.2, 149.2, 137.6, 136.9, 136.6, 130.7, 129.7, 128.6, 128.2, 124.8, 123.0, 35.7, 31.5, 30.9, 28.7, 22.5, 14.4

[0378] Example 1.140: 4-((1H-Indol-2-yl)methylene)-2-(4-(tert-butyl)phenyl)oxazol-5(4H)-one (F140) [Chemical formula]

[0379] 11H NMR (400 MHz, DMSO-d 6 ) δ 11.27 (s, 1H), 8.18 (d, 2H, J = 8.0 Hz), 7.63 (t, 4H, J = 7.4 Hz), 7.35 (d, 2H, J = 8.8 Hz), 7.25 (t, 1H, J = 7.4 Hz), 7.05 (t, 1H, J = 7.6 Hz), 1.30 (s, 9H)

[0380] 13 13C NMR (400 MHz, DMSO-d 6 ) δ 166.9, 161.9, 157.1, 139.7, 133.2, 130.8, 128.5, 128.2, 126.5, 125.6, 122.9, 122.1, 121.0, 120.8, 113.4, 113.0, 35.3, 31.2

[0381] Example 1.141: 4-(Furan-2-ylmethylene)-2-(4-pentylphenyl)oxazol-5(4H)-one (F141)

Chemical Structure

[0382] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.05 (s, 1H), 7.97 (d, 2H, J = 7.6 Hz), 7.56 - 7.55 (m, 1H), 7.40 (d, 2H, J = 7.6 Hz), 7.15 (s, 1H), 6.80 (br s, 1H), 2.64 (t, 2H, J = 7.4 Hz), 1.59 - 1.57 (m, 2H), 1.26 (br s, 4H), 0.82 (t, 3H, J = 6.2 Hz)

[0383] 13 13C NMR (400 MHz, DMSO-d 6 ) δ 167.0, 162.8, 150.4, 149.3, 148.3, 130.5, 129.7, 128.4, 123.0, 120.8, 117.4, 114.6, 35.6, 31.3, 30.7, 22.3, 14.3

[0384] Example 1.142: 2-(4-Butoxyphenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F142)

Chem.

[0385] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 7.98 - 7.96 (m, 3H), 7.78 (d, 1H, J = 3.2 Hz), 7.60 (s, 1H), 7.21 (t, 1H, J = 4.4 Hz), 7.12 (d, 2H, J = 8.4 Hz), 4.05 (t, 2H, J = 6.4 Hz), 1.73 - 1.66 (m, 2H), 1.46 - 1.37 (m, 2H), 0.91 (t, 3H, J = 7.2 Hz)

[0386] 13 C NMR (400 MHz, DMSO-d 6 ) δ 166.8, 163.4, 162.0, 137.7, 136.5, 136.1, 130.9, 130.3, 129.5, 123.8, 117.4, 115.8, 69.2, 31.0, 19.1, 14.1

[0387] Example 1.143: 4-((1H-Pyrrol-2-yl)methylene)-2-(4-propylphenyl)oxazol-5(4H)-one (F143)

Chem.

[0388] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 11.52 (s, 1H), 8.07 (d, 2H, J = 8.0 Hz), 7.39 (d, 2H, J = 8.4 Hz), 7.31 (s, 1H), 7.19 (s, 1H), 7.04 (s, 1H), 6.34 - 6.32 (m, 1H), 2.63 (t, 2H, J = 8.4 Hz), 1.65 - 1.56 (m, 2H), 0.88 (t, 3H, J = 7.4 Hz)

[0389] ESI(m / z) 281 (MH + )

[0390] Example 1.144: 2-(4-Propylphenyl)-4-(thiophen-2-ylmethylene)oxazol-5(4H)-one (F144)

Chem.

[0391] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.02 (d, 1H, J = 5.2 Hz), 7.96 (d, 2H, J = 8.4 Hz), 7.81 (d, 1H, J = 3.6 Hz), 7.67 (s, 1H), 7.42 (d, 2H, J = 8.0 Hz), 7.23 - 7.21 (m, 1H), 2.64 (t, 2H, J = 7.4 Hz), 1.65 - 1.56 (m, 2H), 0.88 (t, 3H, J = 7.4 Hz)

[0392] ESI(m / z) 298 (MH + )

[0393] Example 1.145: 2-(4-Propylphenyl)-4-(thiophen-3-ylmethylene)oxazol-5(4H)-one (F145)

Chem.

[0394] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 8.02 (d, 1H, J = 5.2 Hz), 7.97 (d, 2H, J = 4.4 Hz), 7.81 (d, 1H, J = 3.6 Hz), 7.67 (s, 1H), 7.42 (d, 2H, J = 8.4 Hz), 7.23 - 7.21 (m, 1H), 2.63 (t, 2H, J = 8.4 Hz), 1.65 - 1.56 (m, 2H), 0.88 (t, 3H, J = 7.2 Hz)

[0395] ESI(m / z) 298 (MH+ )

[0396] Example 1.146: 4-(furan-2-ylmethylene)-2-(4-propylphenyl)oxazol-5(4H)-one (F146) [Chemical formula]

[0397] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.05 (s, 1H), 7.98 (d, 2H, J = 8.0 Hz), 7.56 (s, 1H), 7.41 (d, 2H, J = 8.0 Hz), 7.15 (s, 1H), 6.80 (s, 1H), 2.63 (t, 2H, J = 7.4 Hz), 1.63 - 1.57 (m, 2H), 0.87 (t, 3H, J = 7.2 Hz)

[0398] 13 13C HMR (400 MHz, DMSO-d 6 ) δ 167.0, 162.8, 150.4, 149.0, 148.3, 130.5, 129.82, 128.42, 123.0, 120.8, 117.4, 114.6, 37.7, 24.1, 14.0 ESI (m / z) 282 (MH + )

[0399] Example 1.147: 4-((1H-pyrrol-2-yl)methylene)-2-(4-hexylphenyl)oxazol-5(4H)-one (F147) [Chemical formula]

[0400] ESI (m / z) 323 (MH + )

[0401] Example 1.148: 2-(4-(4-((1H-pyrrol-2-yl)methylene)-5-oxo-4,5-dihydrooxazol-2-yl)phenyl)-2-methylpropenenitrile (F148) [Chemical formula]

[0402] ESI(m / z) 306 (MH + )

[0403] Example 1.149: 4 - ((1H - pyrrol - 2 - yl)methylene)-2-(3,5 - dimethylphenyl)oxazol - 5(4H)-one (F149) [Chemical formula]

[0404] ESI(m / z) 267 (MH + )

[0405] Example 1.150: 2 - (3,5 - dimethylphenyl)-4-(thiophen - 2 - ylmethylene)oxazol - 5(4H)-one (F150) [Chemical formula]

[0406] 1 H NMR (400 MHz, MHz, DMSO - d 6 ) δ 8.01 (d, 1H, J = 4.8 Hz), 7.79 (d, 1H, J = 3.2 Hz), 7.65 (s, 1H), 7.63 (s, 1H), 7.28 (s, 1H), 7.21 (t, 1H, J = 4.2 Hz), 2.33 (s, 6H)

[0407] ESI(m / z) 284 (MH + )

[0408] Example 2.1: Methyl 2-(4-(tert - butyl)benzamide)-3-(1H - pyrrol - 2 - yl)acrylate (G1) [Chemical formula]

[0409] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 11.56 (s, 1H), 9.73 (s, 1H), 7.94 (d, 2H, J = 8.1 Hz), 7.50 (d, 2H, J = 8.1 Hz), 7.45 (s, 1H), 6.97 (s, 1H), 6.54 - 6.47 (m, 1H), 6.12 (s, 1H), 3.64 (s, 3H), 1.29 (s, 9H)

[0410] Example 2.2: Methyl 2-(4-(tert-butyl)benzamido)-3-(thiophen-2-yl)acrylate (G2)

Chemical Structure

[0411] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 9.77 (s, 1H), 7.95 (d, 2H, J = 8.3 Hz), 7.88 (s, 1H), 7.70 (d, 1H, J = 5.0 Hz), 7.57 - 7.50 (m, 3H), 7.12 - 7.09 (m, 1H), 3.69 (s, 3H), 1.29 (s, 9H)

[0412] Example 2.3: 2-(4-(tert-butyl)benzamido)-3-(thiophen-2-yl)acrylate (G3)

Chemical Structure

[0413] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 12.62 (s, 1H), 9.64 (s, 1H), 7.93 (d, 2H, J = 8.3 Hz), 7.83 (s, 1H), 7.66 (d, 1H, J = 5.0 Hz), 7.54 - 7.49 (m, 3H), 7.09 (t, 1H, J = 4.0 Hz), 1.29 (s, 9H)

[0414] ESI (m / z) 352 (MNa + )、328 (MH -)

[0415] Example 2.4: Methyl 2-(4-(tert-butyl)benzamide)-3-(furan-2-yl)acrylate (G4)

Chemical Structure

[0416] 2-(4-(tert-Butyl)phenyl)-4-(furan-2-ylmethylene)oxazol-5(4H)-one (F38) (300 mg, 1.02 mmol) was added to MeOH (10 ml) under stirring. When the suspension became a clear solution, triethylamine (0.425 ml, 3.05 mmol) was added to the mixture and stirred for 1 hour. After the reaction was completed (TLC), the solvent was removed under reduced pressure. The solid passed through a filtration funnel and recrystallized from methanol to give 2-(4-(tert-butyl)benzamide)-3-(furan-2-yl)acrylate (G4).

[0417] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 9.87 (s, 1H), 7.92 (d, 2H, J = 8.3 Hz), 7.81 (s, 1H), 7.51 (d, 2H, J = 8.3 Hz), 7.24 (s, 1H), 6.84 (d, 1H, J = 3.4 Hz), 6.60 - 6.57 (m, 1H), 3.68 (s, 3H), 1.29 (s, 9H)

[0418] ESI (m / z) 328 (MH + )、350 (MNa + )、326 (MH - )

[0419] Example 2.5: 2-(4-(tert-Butyl)benzamide)-3-(furan-2-yl)acrylic acid (G5)

Chemical Structure

[0420] 11H NMR (400 MHz, DMSO-d 6 ) δ 12.67 (s, 1H), 9.72 (s, 1H), 7.90 (d, 2H, J = 8.2 Hz), 7.78 (s, 1H), 7.50 (d, 2H, J = 8.3 Hz), 7.23 (s, 1H), 6.78 (d, 1H, J = 3.3 Hz), 6.58 - 6.55 (m, 1H), 1.28 (s, 9H)

[0421] ESI (m / z) 336 (MNa + )、312 (MH - )

[0422] Example 2.6: tert-Butyl 2-(4-(tert-butyl)benzamido)-3-(1H-pyrrol-2-yl)acrylate (G6)

Chemical Structure

[0423] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 11.24 (s, 1H), 9.49 (s, 1H), 7.90 (d, 2H, J = 8.3 Hz), 7.50 (d, 2H, J = 8.3 Hz), 7.33 (s, 1H), 6.94 (s, 1H), 6.48 (s, 1H), 6.10 (s, 1H), 1.39 (s, 9H), 1.29 (s, 9H)

[0424] Example 2.7: 2-(4-(tert-butyl)benzamido)-3-(1H-pyrrol-2-yl)acrylic acid (G7)

Chemical Structure

[0425] 4-((1H-Pyrrol-2-yl)methylene)-2-(4-(tert-butyl)phenyl)oxazol-5(4H)-one (F10) (1.02 mmol) was added to acetone (5 ml) under stirring. When the suspension became a clear solution, 1% NaOH (3.05 mmol) was added to the mixture and stirred for 1 hour. After the reaction was completed (TLC), it was acidified with 10% HCl and then passed through a filtering funnel to give a solid, 2-(4-(tert-butyl)benzamide)-3-(1H-pyrrol-2-yl)acrylic acid (G7).

[0426] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 12.23 (s, 1H), 11.24 (s, 1H), 9.48 (s, 1H), 7.92 (d, 2H, J = 8.4 Hz), 7.50 (d, 2H, J = 8.0 Hz), 7.43 (s, 1H), 6.95 (s, 1H), 6.47 (s, 1H), 6.11 (s, 1H), 1.29 (s, 9H)

[0427] Example 2.8: 2-(4-(tert-Butyl)benzamide)-3-(thiophen-3-yl)acrylic acid (G8)

Chemical formula

[0428] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 12.59 (S, 1H), 9.76 (s, 1H), 7.91 (d, 3H, J = 9.6 Hz), 7.51 (d, 4H, J = 7.2 Hz), 7.37 (d, 1H, J = 4.8 Hz), 1.29 (s, 9H)

[0429] Example 2.9: 2-(4-(tert-Butyl)benzamide)-3-(4-methylthiazol-5-yl)acrylic acid (G9)

Chemical formula

[0430] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 12.83 (s, 1H), 9.73 (s, 1H), 9.00 (s, 1H), 7.92 (d, 2H, J = 8.4 Hz), 7.74 (s, 1H), 7.53 (d, 2H, J = 8.4 Hz), 2.47 (s, 3H), 1.29 (s, 9H)

[0431] Example 2.10: 2-(4-(tert-Butyl)benzamido)-3-(1-methyl-1H-pyrazol-4-yl)acrylic acid (G10)

Chemical Structure

[0432] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 12.39 (s, 1H), 9.57 (s, 1H), 7.95 - 7.88 (m, 3H), 7.61 (s, 1H), 7.51 (d, 2H, J = 8.0 Hz), 7.39 (s, 1H), 3.79 (s, 3H), 1.29 (s, 9H)

[0433] ESI (m / z) 328 (MH + )、350 (MNa + )、327 (MH - )

[0434] Example 2.11: Methyl 2-(4-butylbenzamido)-3-(1H-pyrrol-2-yl)acrylate (G11)

Chemical Structure

[0435] 1 1H NMR (400 MHz, DMSO-d 6) δ 11.35 (s, 1H), 9.64 (s, 1H), 7.95 (d, 2H, J = 6.7 Hz), 7.49 (s, 1H), 7.34 (d, 2H, J = 6.8 Hz), 7.02 (s, 1H), 6.55 (s, 1H), 6.16 (s, 1H), 3.68 (s, 3H), 2.72 - 2.60 (m, 2H), 1.65 - 1.52 (m, 2H), 1.40 - 1.25 (m, 2H), 0.98 - 0.84 (m, 3H)

[0436] ESI(m / z) 327 (MH + )、349 (MNa + )、325 (MH - )

[0437] Example 2.12: 2-(4-Butylbenzamido)-3-(thiophen-2-yl)acrylic acid (G12)

Chemical Structure

[0438] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 12.67 (s, 1H), 9.67 (s, 1H), 7.95 (d, 2H, J = 7.9 Hz), 7.87 (s, 1H), 7.70 (d, 1H, J = 4.9 Hz), 7.59 - 7.49 (m, 1H), 7.36 (d, 2H, J = 7.9 Hz), 7.13 (d, 1H, J = 4.4 Hz), 2.67 (t, 2H, J = 7.6 Hz), 1.66 - 1.55 (m, 2H), 1.40 - 1.26 (m, 2H), 0.91 (t, 3H, J = 7.3 Hz)

[0439] ESI(m / z) 330 (MH + )、328 (MH - )

[0440] Example 2.13: 2-(4-(tert-Butyl)benzamido)-3-(1-phenyl-1H-pyrrol-2-yl)acrylic acid (G13)

Chemical Structure

[0441] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 12.39 (s, 1H), 9.70 (s, 1H), 7.98 (d, 2H, J = 7.8 Hz), 7.65 - 7.49 (m, 5H), 7.38 (d, 2H, J = 7.4 Hz), 7.23 (s, 1H), 7.16 (s, 1H), 6.78 (s, 1H), 6.34 (s, 1H), 1.33 (s, 9H)

[0442] ESI (m / z) 389 (MH + )、387 (MH - )

[0443] Example 2.14: Methyl 2-(4-(tert-butyl)benzamide)-3-(1-phenyl-1H-pyrrol-2-yl)acrylate (G14)

Chemical Structure

[0444] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 9.81 (s, 1H), 7.96 (d, 2H, J = 7.7 Hz), 7.62 - 7.45 (m, 5H), 7.35 (d, 2H, J = 7.3 Hz), 7.22 (s, 1H), 7.11 (s, 1H), 6.78 (s, 1H), 6.33 (s, 1H), 3.58 (s, 3H), 1.30 (s, 9H)

[0445] ESI (m / z) 403 (MH + )、425 (MNa + )、401 (MH - )

[0446] Example 2.15: Methyl 3-(1H-pyrrol-2-yl)-2-(4-(trifluoromethyl)benzamide)acrylate (G15)

Chemical Structure

[0447] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 11.37 (s, 1H), 9.98 (s, 1H), 8.22 (d, 2H, J = 8.1 Hz), 7.93 (d, 2H, J = 8.2 Hz), 7.53 (s, 1H), 7.04 (s, 1H), 6.56 (s, 1H), 6.18 (s, 1H), 3.70 (s, 3H) ESI (m / z) 339 (MH + )、361 (MNa + )、337 (MH - )

[0448] Example 2.16: 2-(4-Bromobenzamido)-3-(furan-2-yl)acrylic acid (G16)

Chem.

[0449] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 12.78 (s, 1H), 9.93 (s, 1H), 7.94 (d, 2H, J = 8.2 Hz), 7.82 (s, 1H), 7.75 (d, 2H, J = 8.4 Hz), 7.30 (s, 1H), 6.84 (d, 1H, J = 3.3 Hz), 6.64 - 6.58 (m, 1H)

[0450] ESI (m / z) 336 (MH + )、338 (MH + )、334 (MH - )、335 (MH - )

[0451] Example 2.17: Methyl 3-(furan-2-yl)-2-(4-(trifluoromethyl)benzamido)acrylate (G17)

Chem.

[0452] 1 H NMR (400 MHz, MHz, DMSO-d6 ) δ 10.23 (s, 1H), 8.19 (d, 2H, J = 8.0 Hz), 7.93 (d, 2H, J = 8.2 Hz), 7.88 - 7.86 (m, 1H), 7.35 (s, 1H), 6.93 (d, 1H, J = 3.4 Hz), 6.66 - 6.62 (m, 1H), 3.73 (s, 3H)

[0453] ESI (m / z) 340 (MH + )、362 (MNa + )、338 (MH - )

[0454] Example 2.18: 3-(Furan-2-yl)-2-(4-(trifluoromethyl)benzamide)acrylic acid (G18)

Chemical Structure

[0455] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 12.83 (s, 1H), 10.10 (s, 1H), 8.19 (d, 2H, J = 7.9 Hz), 7.93 (d, 2H, J = 8.2 Hz), 7.83 (s, 1H), 7.34 (s, 1H), 6.87 (d, 1H, J = 3.3 Hz), 6.63 - 6.59 (m, 1H)

[0456] ESI (m / z) 326 (MH + )、324 (MH - )

[0457] Example 2.19: 2-(4-Bromobenzamide)-3-(thiophen-2-yl)acrylic acid (G19)

Chemical Structure

[0458] 1 H NMR (400 MHz, MHz, DMSO-d 6) δ 12.74 (s, 1H), 9.87 (s, 1H), 7.98 (d, 2H, J = 8.5 Hz), 7.90 (s, 1H), 7.78 (d, 2H, J = 8.5 Hz), 7.72 (d, 1H, J = 5.1 Hz), 7.56 (d, 1H, J = 3.3 Hz), 7.16 - 7.12 (m, 1H)

[0459] ESI (m / z) 354 (MH + )、352 (MH + )、352 (MH - )、350 (MH - )

[0460] Example 2.20: Methyl 2-(4-bromobenzamide)-3-(furan-2-yl)acrylate (G20)

Chemical Structure

[0461] 1 1H NMR (400 MHz, MHz, DMSO-d 6 ) δ 10.03 (s, 1H), 7.91 (d, 2H, J = 8.5 Hz), 7.82 (d, 1H, J = 1.5 Hz), 7.72 (d, 2H, J = 8.5 Hz), 7.27 (s, 1H), 6.86 (d, 1H, J = 3.5 Hz), 6.61 - 6.58 (m, 1H), 3.69 (s, 3H)

[0462] ESI (m / z) 350 (MH + )、352 (MH + )、348 (MH - )、350 (MH - )

[0463] Example 2.21: 2-(4-bromobenzamide)-3-(furan-2-yl)acrylic acid (G21)

Chemical Structure

[0464] 1 1H NMR (400 MHz, MHz, DMSO-d 6) δ 12.78 (s, 1H), 9.93 (s, 1H), 7.94 (d, 2H, J = 8.5 Hz), 7.82 (d, 1H, J = 1.5 Hz), 7.75 (d, 2H, J = 8.5 Hz), 7.31 (s, 1H), 6.84 (d, 1H, J = 3.4 Hz), 6.63 - 6.59 (m, 1H)

[0465] ESI (m / z) 336 (MH + )、338 (MH + )、334 (MH - )、336 (MH - )

[0466] Example 2.22: 2-(4-Butylbenzamide)-3-(1H-pyrrol-2-yl)acrylic acid (G22)

Chemical Structure

[0467] 1 1H NMR (400 MHz, MHz, DMSO-d 6 ) δ 12.28 (s, 1H), 11.28 (s, 1H), 9.51 (s, 1H), 7.94 (d, 2H, J = 8.0 Hz), 7.47 (s, 1H), 7.33 (d, 2H, J = 8.0 Hz), 6.99 (s, 1H), 6.51 (s, 1H), 6.14 (d, 1H, J = 2.7 Hz), 2.66 (t, 2H, J = 7.6 Hz), 1.64 - 1.54 (m, 2H), 1.38 - 1.27 (m, 2H), 0.91 (t, 3H, J = 7.3 Hz)

[0468] Example 2.23: Methyl-2-(4-(tert-butyl)benzamide)-3-(1H-pyrrol-2-yl)acrylate (G23)

Chemical Structure

[0469] 1 1H NMR (400 MHz, MHz, DMSO-d 6) δ 11.31 (s, 1H), 9.61 (s, 1H), 7.94 (d, 2H, J = 8.0 Hz), 7.51 (d, 2H, J = 8.0 Hz), 7.46 (s, 1H), 6.98 (s, 1H), 6.52 (s, 1H), 6.13 (s, 1H), 3.65 (s, 3H), 1.29 (s, 9H) 13 C NMR (400 MHz, DMSO-d 6 ) δ 166.1, 165.9, 154.9, 131.4, 128.0, 126.6, 126.5, 125.6, 122.6, 119.7, 113.9, 111.0, 52.2, 35.1, 31.4

[0470] Example 2.24: 2-(2-Naphthamide)-3-(thiophen-3-yl)acrylic acid (G24)

Chemical formula

[0471] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 12.68 (s, 1H), 10.03 (s, 1H), 8.63 (s, 1H), 8.05 - 8.03 (m, 3H), 7.98 (d, 1H, J = 7.6 Hz), 7.94 (br s, 1H), 7.64 - 7.59 (m, 2H), 7.56 (s, 1H), 7.53 - 7.52 (m, 1H), 7.41 (d, 1H, J = 4.4 Hz)

[0472] 13 C NMR (400 MHz, DMSO-d 6 ) δ 166.9, 166.4, 135.8, 134.8, 132.6, 131.3, 130.3, 129.4, 128.7, 128.5, 128.3, 128.2, 128.1, 127.3, 127.2, 125.9, 124.7

[0473] Example 2.25: 3-(Benzo[b]thiophen-3-yl)-2-(4-(tert-butyl)benzamide)acrylic acid (G25)

Chemical formula

[0474] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 12.78 (s, 1H), 9.90 (s, 1H), 8.10 (s, 1H), 8.01 (d, 1H, J = 7.6 Hz), 7.96 (d, 1H, J = 7.6 Hz), 7.91 (d, 2H, J = 8.0 Hz), 7.67 (s, 1H), 7.50 (d, 2H, J = 8.0 Hz), 7.46 - 7.38 (m, 2H), 1.28 (s, 9H)

[0475] 13 13C NMR (400 MHz, DMSO-d 6 ) δ 166.6, 166.3, 139.2, 138.5, 131.2, 129.5, 129.0, 128.9, 128.1, 125.6, 125.4, 125.2, 123.9, 123.4, 131.9, 35.1, 31.4

[0476] Example 2.26: 2-(4-Isobutylbenzamide)-3-(1H-pyrrol-2-yl)acrylic acid (G26) [Chemical formula]

[0477] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 12.26 (br s, 1H), 11.25 (s, 1H), 9.48 (s, 1H), 7.91 (d, 2H, J = 7.2 Hz), 7.44 (s, 1H), 7.27 (d, 2H, J = 7.6 Hz), 6.95 (s, 1H), 6.50 (s, 1H), 6.11 (s, 1H), 2.50 (d, 2H, J = 2.8 Hz), 1.90 - 1.91 (m, 1H), 0.85 (d, 6H, J = 6.8 Hz)

[0478] 13 13C NMR (400 MHz, DMSO-d 6)δ 167.0, 165.9, 145.4, 132.0, 129.3, 129.0, 126.7, 126.1, 122.2, 120.7, 113.4, 110.8, 44.8, 30.0, 22.57

[0479] Example 2.27: 2-(1-Naphthamide)-3-(4-methylthiazol-5-yl)acrylic acid (G27) [Chemical formula]

[0480] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 12.98 (br s, 1H), 9.97 (s, 1H), 9.09 (s, 1H), 8.40 - 8.39 (m, 1H), 8.6 (d, 1H, J = 8.0 Hz), 7.99 - 7.98 (m, 1H), 7.89 (d, 1H, J = 7.2 Hz), 7.80 (s, 1H), 7.62 (t, 1H, J = 7.6 Hz), 7.57 - 7.55 (m, 2H), 2.54 (s, 3H)

[0481] 13 C NMR (400 MHz, DMSO-d 6 ) δ 169.3, 166.1, 156.8, 156.6, 134.1, 133.6, 130.8, 130.3, 128.6, 127.3, 126.8, 126.2, 126.1, 126.0, 125.9, 125.4, 124.9, 16.1

[0482] Example 2.28: 2-(4-Propylbenzamide)-3-(1H-pyrrol-2-yl)acrylic acid (G28) [Chemical formula]

[0483] 1 H NMR (400 MHz, MHz, DMSO-d 6) δ 12.30 (broad s, 1H), 11.25 (s, 1H), 9.48 (s, 1H), 7.91 (d, 2H, J = 7.6 Hz), 7.44 (s, 1H), 7.30 (d, 2H, J = 4.0 Hz), 6.95 (s, 1H), 6.48 (s, 1H), 6.11 (s, 1H), 2.60 (t, 2H, J = 7.2 Hz), 1.64 - 1.55 (m, 2H), 0.87 (t, 3H, J = 7.0 Hz)

[0484] 13 C NMR (400 MHz, DMSO-d 6 ) δ 167.0, 165.9, 146.4, 131.9, 129.7, 128.1, 126.7, 126.1, 122.1, 120.7, 113.4, 110.8, 37.5, 24.3, 14.0

[0485] Example 2.29: 2-(4-Butylbenzamido)-3-(thiophen-2-yl)acrylic acid (G29)

Chemical Structure

[0486] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 12.61 (s, 1H), 9.76 (s, 1H), 7.89 (s, 3H), 7.54 - 7.52 (m, 1H), 7.51 (s, 1H), 7.37 (d, 1H, J = 4.8 Hz), 7.31 (d, 2H, J = 7.6 Hz), 2.62 (t, 2H, J = 7.6 Hz), 1.59 - 1.51 (m, 2H), 1.33 - 1.24 (m, 2H), 0.87 (t, 3H, J = 7.2 Hz)

[0487] ESI (m / z) 330 (MH + )

[0488] Example 2.30: 2-(4-Propylbenzamido)-3-(thiophen-2-yl)acrylic acid (G30)

Chemical Structure

[0489] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 12.64 (s, 1H), 9.65 (s, 1H), 7.93 (d, 2H, J = 8.0 Hz), 7.85 (s, 1H), 7.66 (d, 1H, J = 4.4 Hz), 7.51 (d, 1H, J = 2.0 Hz), 7.32 (d, 2H, J = 8.0 Hz), 7.10 (d, 1H, J = 3.6 Hz), 2.60 (t, 2H, J = 7.4 Hz), 1.63 - 1.56 (m, 2H), 0.88 (t, 3H, J = 7.2 Hz)

[0490] 13 13C HMR (400 MHz, DMSO-d 6 ) δ 166.5, 166.4, 146.7, 137.0, 133.8, 131.8, 131.7, 129.9, 128.8(2), 128.2(2), 127.5, 124.5, 37.5, 24.3, 14.1 ESI (m / z) 316 (MH + )

[0491] Example 2.31: 2-(4-Propylbenzamide)-3-(thiophen-2-yl)acrylic acid (G31)

Chemical Structure

[0492] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 12.64 (s, 1H), 9.64 (s, 1H), 7.92 (d, 2H, J = 7.6 Hz), 7.84 (s, 1H), 7.66 (d, 1H, J = 4.4 Hz), 7.51 (s, 1H), 7.32 (d, 2H, J = 7.6 Hz), 7.10 - 7.09 (m, 1H), 2.60 (t, 2H, J = 7.4 Hz), 1.63 - 1.57 (m, 2H), 0.88 (t, 3H, J = 7.6 Hz)

[0493] 13 13C HMR (400 MHz, DMSO-d 6)δ166.5, 166.4, 146.7, 136.9, 133.8, 131.8, 131.7, 129.8, 128.8(2), 128.2(2), 127.5, 124.5, 37.5, 24.4, 14.1

[0494] ESI(m / z) 316 (MH + )

[0495] Example 2.32: 2-(4-(tert-Butyl)benzamide)-3-(4-methylthiazol-5-yl)acrylic acid (G32)

Chemical Structure

[0496] ESI(m / z) 345 (MH + )

[0497] Example 2.33: 2-(4-(tert-Butyl)benzamide)-3-(1H-indol-2-yl)acrylic acid (G33)

Chemical Structure

[0498] ESI(m / z) 363 (MH + )

[0499] Example 2.34: 3-(Furan-2-yl)-2-(4-propylbenzamide)acrylic acid (G34)

Chemical Structure

[0500] 1 1H NMR (400 MHz, MHz, DMSO-d 6)δ 12.69 (s, 1H), 9.72 (s, 1H), 7.89 (d, 2H, J = 7.6 Hz), 7.78 (s, 1H), 7.30 (d, 2H, J = 7.6 Hz), 7.24 (s, 1H), 6.78 (d, 1H, J = 2.4 Hz), 6.57 (s, 1H), 2.60 (t, 2H, J = 7.4 Hz), 1.64 - 1.55 (m, 2H), 0.87 (t, 3H, J = 7.2 Hz)

[0501] 13 C HMR (400 MHz, DMSO-d 6 )δ 166.4, 165.8, 149.8, 146.7, 145.6, 131.6, 128.8 (2), 128.2 (2), 124.9, 120.9, 115.7, 112.9, 37.5, 24.3, 14.0

[0502] ESI (m / z) 300 (MH + )

[0503] Example 2.35: 3-(Furan-2-yl)-2-(4-pentylbenzamide) acrylic acid (G35)

Chemical formula

[0504] 1 H NMR (400 MHz, MHz, DMSO-d 6 )δ 12.68 (s, 1H), 9.72 (s, 1H), 7.89 (d, 2H, J = 7.6 Hz), 7.78 (s, 1H), 7.30 (d, 2H, J = 7.2 Hz), 7.24 (s, 1H), 6.78 (d, 1H, J = 3.2 Hz), 6.57 (t, 1H, J = 1.4 Hz), 2.61 (t, 2H, J = 7.4 Hz), 1.61 - 1.55 (m, 2H), 1.26 (s, 4H), 0.83 (t, 3H, J = 6.8 Hz)

[0505] 13 C HMR (400 MHz, DMSO-d 6) δ 166.4, 165.8, 149.8, 146.9, 145.6, 131.6, 128.7(2), 128.2(2), 124.9, 120.9, 115.7, 112.9, 35.4, 31.3, 30.8, 22.4, 14.3

[0506] ESI(m / z) 328 (MH + )

[0507] Example 2.36: 2-(4-Butoxybenzamide)-3-(thiophen-2-yl)acrylic acid (G36)

Chemical Structure

[0508] 1 H NMR (400 MHz, DMSO-d 6 ) δ 12.61 (s, 1H), 9.55 (s, 1H), 7.96 (d, 2H, J = 8.4 Hz), 7.82 (s, 1H), 7.66 (d, 1H, J = 4.8 Hz), 7.50 (d, 1H, J = 2.4 Hz), 7.09 (t, 1H, J = 4.2 Hz), 7.03 (d, 2H, J = 8.4 Hz), 4.03 (t, 2H, J = 6.4 Hz), 1.73 - 1.66 (m, 2H), 1.47 - 1.37 (m, 2H), 0.91 (t, 3H, J = 7.4 Hz)

[0509] 13 C HMR (400 MHz, DMSO-d 6 ) δ 166.5, 166.1, 161.9, 137.0, 133.7, 131.7, 130.1(2), 129.7, 127.4, 126.3, 124.7, 114.5(2), 67.9, 31.1, 19.1, 14.1

[0510] ESI(m / z) 346 (MH + )

[0511] Example 2.37: 2-(4-Hexylbenzamide)-3-(thiophen-2-yl)acrylic acid (G37)

Chemical Structure

[0512] ESI(m / z) 358 (MH + )

[0513] Example 2.38: 2-(2,3-Dihydro-1H-inden-5-carboxamido)-3-(thiophen-2-yl)acrylic acid (G38)

Chem.

[0514] 1 1H NMR (400 MHz, MHz, DMSO-d 6 ) δ 12.62 (s, 1H), 9.59 (s, 1H), 7.84 (d, 2H, J = 7.4 Hz), 7.78 (d, 1H, J = 7.6 Hz), 7.66 (d, 1H, J = 4.0 Hz), 7.51 (br s, 1H), 7.33 (d, 1H, J = 7.6 Hz), 7.10 - 7.09 (m, 1H), 2.90 (t, 4H, J = 6.8 Hz), 2.05 - 2.01 (m, 2H)

[0515] ESI(m / z) 314 (MH + )

[0516] Example 2.39: 2-(4-(Difluoromethoxy)benzamido)-3-(thiophen-2-yl)acrylic acid (G39)

Chem.

[0517] 1 1H NMR (400 MHz, MHz, DMSO-d 6 ) δ 12.69 (s, 1H), 9.77 (s, 1H), 8.07 (d, 2H, J = 8.4 Hz), 7.86 (s, 1H), 7.68 (d, 1H, J = 4.4 Hz), 7.55 - 7.53 (m, 1H), 7.36 (s, 1H), 7.31 (d, 2H, J = 8.4 Hz), 7.10 (t, 1H, J = 3.8 Hz)

[0518] ESI(m / z) 340 (MH + )

[0519] Example 2.40: 2-(4-Pentylbenzamido)-3-(thiophen-2-yl)acrylic acid (G40)

Chemical Structure

[0520] 1 H NMR (400 MHz, MHz, DMSO-d 6 ) δ 12.63 (s, 1H), 9.64 (s, 1H), 7.91 (d, 2H, J = 8.4 Hz), 7.83 (s, 1H), 7.66 (d, 1H, J = 4.8 Hz), 7.51 (d, 1H, J = 2.8 Hz), 7.32 (d, 2H, J = 8.0 Hz), 7.10 - 7.08 (m, 1H), 2.62 (t, 2H, J = 8.0 Hz), 1.61 - 1.54 (m, 2H), 1.31 - 1.23 (m, 4H), 0.83 (t, 3H, J = 8.0 Hz)

[0521] 13 C HMR (400 MHz, DMSO-d 6 ) δ 166.5, 166.4, 146.9, 136.9, 133.8, 131.8, 131.7, 129.8, 128.7(2), 128.2(2), 127.5, 124.5, 35.4, 31.3, 30.8, 22.4, 14.3

[0522] ESI(m / z) 344 (MH + )

[0523] Example 2.41: 2-(4-Butoxybenzamido)-3-(1H-pyrrol-2-yl)acrylic acid (G41)

Chemical Structure

[0524] 1 H NMR (400 MHz, MHz, DMSO-d 6)δ 12.21 (s, 1H), 11.22 (s, 1H), 9.37 (s, 1H), 7.94 (d, 2H, J = 8.4 Hz), 7.41 (s, 1H), 6.99 (d, 2H, J = 8.4 Hz), 6.94 (s, 1H), 6.45 (s, 1H), 6.09 (s, 1H), 4.02 (t, 2H, J = 8.3 Hz), 1.72 - 1.65 (m, 2H), 1.46 - 1.37 (m, 2H), 0.90 (t, 3H, J = 7.4 Hz)

[0525] ESI (m / z) 329 (MH + )

[0526] Example 2.42: 2-(3,5-Dimethylbenzamide)-3-(thiophen-2-yl)acrylic acid (G42)

Chem.

[0527] 1 1H NMR (400 MHz, MHz, DMSO-d 6 )δ 12.62 (s, 1H), 9.60 (s, 1H), 7.83 (s, 1H), 7.67 (d, 1H, J = 4.8 Hz), 7.61 (s, 2H), 7.50 (d, 1H, J = 2.0 Hz), 7.19 (s, 1H), 7.10 - 7.08 (m, 1H), 2.32 (s, 6H)

[0528] ESI (m / z) 302 (MH + )

[0529] Example 3: Function of New Compounds as Pendrin Inhibitors Example 3.1: Cell Culture Chinese hamster ovary (CHO)-K1 cells were maintained in DMEM medium containing 10% fetal bovine serum (FBS), 100 units / ml penicillin and 100 μg / ml streptomycin. CHO-K1 cells were stably transfected with pcDNA3.1 encoding halide sensor YFP-H148Q / I152L / F46L and human wild-type (WT)-pendrin. For the differentiation of primary cultures of human nasal epithelial (HNE) cells, passage-2 cells were 2×105 cells / cm 2 Cells were seeded onto Transwell-clear culture inserts with a pore size of 0.45 μm (Costar Co., Cambridge, MA) at a density of. Cells were maintained in a 1:1 mixture of Dulbecco’s modified Eagle’s medium (Lonza, Walkersville, MD) and bronchial epithelial growth medium (Lonza) supplemented with the following growth factors according to the manufacturer's instructions. After immersing the cells for the first 7 days, they were exposed to the apical air interface for the remainder of the culture period. Cells were used between 14 and 21 days after the establishment of the air-liquid interface. At all stages of the culture, the cells were maintained at 37 °C in a 5% CO 2 incubator with air.

[0530] Example 3.2: Cell-Based High-Throughput Screening CHO-K1 cells expressing human WT pendrin and YFP-F46L / H148Q / I152L were plated in 96-well microplates at a density of 2×10 4 cells per well and cultured for 48 hours. Each well of the 96-well microplate in which the cells were cultured was washed twice with 200 μL of PBS and filled with 50 μL of HEPES buffer solution each. A test compound (1 μL) was added at a final concentration of 50 μM. After culturing at 37 °C for 10 minutes, the 96-well microplate was placed in a FLUOstar Omega microplate reader (BMG Labtech, Ortenberg, Germany) for fluorescence analysis. By continuously recording fluorescence for 1 second (baseline) (400 ms per point), each well was individually analyzed for pendrin-mediated I - influx. Then, 50 μL of NaI-substituted HEPES buffer solution (NaI replacing NaCl) was added using a liquid injector in 1 second, and YFP fluorescence was recorded for 5 seconds. The initial iodide influx rate was determined from the initial slope of the fluorescence by non-linear regression after injecting iodide (see Figure 1).

[0531] Example 3.3: Measurement of Short Circuit Current Snapwell inserts containing primary cultures of ANO1- and CFTR-expressing FRT and human nasal epithelial (HNE) cells were mounted onto an Ussing chamber (Physiologic Instruments, San Diego, CA, USA).

[0532] To measure the short-circuit current in HNE cells, the apical and basal chambers were filled with symmetrical HCO 3 - buffer solution. For ANO1- and CFTR-expressing FRT, the apical chamber was filled with half-Cl - solution and the side chamber was filled with HCO 3 - buffer solution. The basal membrane was permeabilized with 250 μg / mL amphotericin B to measure the apical membrane currents of ANO1 and CFTR. The short-circuit current and apical membrane current were measured with an EVC4000 multi-channel V / I clamp (World Precision Instruments, Sarasota, FL, USA) and a PowerLab4 / 35 (AD Instruments, Castle Hill, Australia). Data were recorded and analyzed using Labchart Pro7 (AD Instruments). The sampling rate was 4 Hz.

[0533] Example 3.4: Measurement of Cl- / I- Exchange Activity Cl - / I - For Cl / I exchange activity measurements, YFP fluorescence in pendrin- and halide sensor YFP-expressing CHO-K1 cells was measured. Each well of the 96-well plate in which the cells were cultured was washed twice with 200 μL of PBS and 50 μL of HEPES buffer solution (140 mM NaCl, 5 mM KCl, 1 mM MgCl 2 、1 mM CaCl 2, filled with 10 mM glucose and 10 mM HEPES (pH 7.4), respectively. A test compound (1 μL) was added at a final concentration of 50 μM. After incubation at 37 °C for 10 minutes, the 96-well plate was placed on a FLUOstar Omega microplate reader (BMG Labtech, Ortenberg, Germany) for fluorescence analysis. By continuously recording fluorescence for 1 second (baseline) (400 ms per point), for pendrin-mediated I - After individually analyzing each well for the influx of, a 50 μL NaI-substituted HEPES buffer solution (140 mM NaI, 5 mM KCl, 1 mM MgCl 2 , 1 mM CaCl 2 , 10 mM glucose and 10 mM HEPES (pH 7.4)) was added and YFP fluorescence was recorded for 5 seconds. The initial iodide influx rate was determined from the initial slope of fluorescence by non-linear regression after injecting iodide.

[0534] Example 3.5: Measurement of Cl- / HCO3- Exchange Activity Cl - / HCO 3 - For measurement of exchange activity, intracellular pH (pH i ) was measured in WT-pendrin-expressing CHO-K1 and HNE cells using the pH sensor SNARF5-AM (Molecular Probes). After treating the cells with 5 μM SNARF5-AM for 30 minutes, they were placed in a perfusion chamber on a reverse fluorescence microscope (Nikon, Tokyo, Japan) equipped with a cooled charge-coupled device camera (Zyla sCMOS) at the stage of image collection and analysis software (Meta Imaging Series 7.7). The cells were perfused with a HCO 2 , 1 CaCl 2 , 10 D-glucose, 5 HEPES, and 25 NaHCO 3 (pH 7.4) (in mM) containing buffer solution. Cl 3 - - / HCO 3 ​- To measure the exchange activity, HCO 3 - buffer solution was changed to Cl - -free HCO 3 - buffer solution. Applying the external Cl - -free HCO 3 - buffer solution increases the efflux of Cl - and the influx of HCO 3 - through pendrin. To maintain the pH of the HCO 3 - buffer solution, the solution was continuously gassed with 95% O 2 and 5% CO 2 . SNARF5 fluorescence was recorded at an excitation wavelength of 515 ± 10 nm and an emission wavelength of 640 ± 10 nm, and intracellular pH correction was performed with a solution containing 145 mM KCl, 10 mM HEPES, and 5 mM nigericin having a pH corrected to 6.2 - 7.6.

[0535] Example 3.6: Measurement of Cl- / SCN- Exchange Activity Cl - / SCN - For the measurement of the exchange activity, YFP fluorescence in pendrin- and halide sensor YFP-expressing CHO-K1 cells was measured. Cl - / SCN - To measure the exchange activity, the HEPES buffer solution was changed to a NaSCN-substituted HEPES buffer solution (140 mM NaSCN, 5 mM KCl, 1 mM MgCl - that drives the influx of SCN - through pendrin (1 mM CaCl 2 , 1 mM CaCl 2 , 10 mM glucose, and 10 mM HEPES (pH 7.4)). The change in YFP fluorescence due to the influx of SCN - was monitored using a FLUOstar Omega microplate reader (BMG Labtech) and MARS data analysis software (BMG Labtech).

[0536] Example 3.7: Measurement of Cl- / OH- Exchange Activity Cl - / OH - For the measurement of exchange activity, intracellular pH (pH i ) was measured in WT-pendrin-expressing CHO-K1 and HNE cells using the pH sensor SNARF5-AM (Molecular Probes). After treating the cells with 5 μM SNARF5-AM for 30 minutes, they were placed in a perfusion chamber on a reverse fluorescence microscope (Nikon, Tokyo, Japan) equipped with a cooled charge-coupled device camera (Zyla sCMOS) and image acquisition and analysis software (Meta Imaging Series 7.7). Cl - / OH - To measure the exchange activity, the HEPES buffer solution was changed to a Cl - -free HEPES buffer solution that drives the efflux of Cl - from the cytosol through pendrin and the influx of OH - to generate a Cl - gradient.

[0537] Example 3.8: Measurement of Protein Expression Level CHO-K1 and HNE cells were lysed in cell lysis buffer (50 mM Tris-HCl, pH 7.4, 1% Nonidet P-40, 0.25% sodium deoxycholate, 150 mM NaCl, 1 mM EDTA, 1 mM Na 3 VO 4and dissolved in a protease inhibitor mixture). The whole cell lysate was centrifuged at 15,000×g for 10 minutes at 4°C to remove cell debris, and the same amount (50 μg of protein or 5 μg of protein / lane) of the supernatant protein was separated on a 4-12% Tris-glycine precast gel (KOMA BIOTECH, Seoul, Korea) and transferred to a PVDF membrane (Millipore, Billerica, MA, USA). The membrane was blocked with 5% non-fat dry milk in TBS containing 0.1% Tween20 (TBST) or 5% bovine serum albumin in TBS for 1 hour at room temperature. Subsequently, the membrane was incubated overnight at 4°C with primary antibodies against pendrin (sc-50346; Santa Cruz Biotechnology, Santa Cruz, CA, USA), NF-κB p65 (4764S; Cell Signaling Technology, Danvers, MA, USA), p-NF-κB p65 (3033S; Cell Signaling Technology), IkBα (9242S; Cell Signaling Technology), p-IkBα (9141S; Cell Signaling Technology), β-actin (sc-47778; Santa Cruz Biotechnology), or ANO1 (ab64085; Abcam). After washing with 0.05% Tween20 in PBS (TBST), the blot was incubated with secondary antibodies (Cell Signaling Technology, Danvers, MA) for 60 minutes at room temperature. Then, the membrane was washed 3 times with TBST for 5 minutes and visualized using an ECL Plus Western blotting detection system (GE Healthcare Amersham; Piscataway, NJ, USA).

[0538] Example 3.9: Real-Time RT-PCR Analysis Total messenger RNA (mRNA) was extracted using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) and reverse-transcribed using random hexamer primers, oligo(dT) primers, and SuperScript® III Reverse Transcriptase (Invitrogen). Quantitative real-time PCR was performed using a StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) and Thunderbird SYBR qPCR mix (Toyobo, Osaka, Japan). Thermal cycling conditions included an initial step of 95°C for 5 minutes in a 96-well reaction plate, followed by 40 cycles of 95°C for 10 seconds, 55°C for 20 seconds, and 72°C for 10 seconds. Primer sequences were listed in Table 3 below.

[0539]

Table 3

[0540] As a result, compounds showing pendrin inhibitory activity at 30 μM are shown in Table 4 below, and IC 50 against compounds showing pendrin inhibitory activity of 50% or more at 30 μM is shown in Table 5 below.

[0541]

Table 4-1

Table 4-2

Table 4-3

[0542]

Table 5

[0543] The pendrin inhibitory effect of F56 was measured in CHO-K1 cells overexpressing WT-pendrin and iodide-sensitive YFP. This significantly inhibited the Cl - / I - exchange activity of pendrin in a dose-dependent manner (Figure 2).

[0544] The inventors also analyzed whether F56 could inhibit the Cl - / HCO 3 - exchange activity. When CHO-K1 cells overexpressing WT-pendrin and iodide-sensitive YFP were treated with a high-concentration Cl - solution, Cl - moved into the cells, and HCO 3 - moved out of the cells. Such changes induced intracellular acidification along with a decrease in YFP fluorescence. Consequently, this significantly inhibited the Cl - / HCO 3 - exchange activity in a dose-dependent manner (Figure 3).

[0545] Cl - / I - Similar to the results of the Cl - / HCO 3 - exchange experiment, F56 strongly inhibited the Cl - / I - and Cl - / HCO 3 - exchange activities. F56 showed 50% inhibition of the Cl - / I - and Cl - / HCO 3 - exchange activities at ~3 μM and ~10 μM, respectively (Figure 4(a)A).

[0546] On the other hand, F10 showed very high activity compared to F56 and inhibited Cl - / I - , Cl - / SCN - , Cl - / HCO 3 - , Cl - / OH -It was confirmed that anion exchange was strongly suppressed (inhibitory effect of F10 on Cl− / I− exchange activity: IC 50 =~80 nM). Therefore, it was confirmed that F10 is a very potent substance among pendrin inhibitors (Figure 4(b)). In addition, G7 is a metabolite of F10, which has lower potency than F10 but is excellent in metabolic stability.

[0547] Example 4: Function of New Compound (F56) in Reducing Airway Hypersensitivity and Mucin Expression in Asthmatic Mice Example 4.1: Thallium Flux Analysis HEK-293T cells were stably transfected with the hERG (human ether-a-go-go-related) gene and seeded at a density of 7×10 4 cells per well in a poly-L-lysine-coated 96-well plate, and the cells were cultured for 48 hours. Four hours before analysis, the cells were switched from 37°C to 28°C for enhanced membrane expression of the hERG channel. Four hours later, the medium was switched to 80 μL / well of FluxOR (Invitrogen) loading buffer, and the cells were cultured for 1 hour in the dark at 37°C. The loading buffer was removed, and 100 μL of assay buffer was added to each well. To measure the effect of F56 on the hERG channel, the cells were pretreated with F56 for 10 minutes. Four hours before adding the stimulation buffer containing 20 μL of thallium ion, FluxOR fluorescence (excitation / emission: 490 / 525 nm) was recorded, and the fluorescence was monitored for an additional 56 seconds. FluxOR fluorescence was recorded and analyzed using a FLUOstar Omega microplate reader (BMG Labtech, Ortenberg, Germany) and MARS data analysis software (BMG Labtech).

[0548] Example 4.2: Measurement of 5-HT2A Activity Human 5-HT 2A , ANO1(abc) and FRT cells expressing YFP-F46L / H148Q / I152L were seeded at 2×10 4Plated in a 96-well microplate at a cell density and cultured for 48 hours. Each well of the 96-well plate was washed twice with 200 μL of PBS and filled with 100 μL of PBS. 5-HT 2A To measure the effect of F56 on the 2A 5-HT - channel, the cells were pretreated with F56. After culturing at 37 °C for 10 minutes, the 96-well plate was placed on a FLUOstar Omega microplate reader for YFP fluorescence measurement. By continuously recording the YFP fluorescence for 2 seconds (800 ms per point), the 5-HT - -mediated influx into each well was analyzed individually (baseline). Then, 100 μL of a 140 mM I

[0549] Example 4.3: Measurement of the Volume of ASL and Fluid Meniscus solution containing 20 μM 5-HT (Sigma-Aldrich) was added in 2 seconds, and then the YFP fluorescence was recorded for 10 seconds. The total volume of the fluid, ASL and the fluid meniscus, together with the cell culture insert wall, was measured by fluid absorption through filter paper. Whatman filter paper (10 mm diameter circle; GE Healthcare) was placed on the top side of a 12-well Transwell insert (Costar Co.) for 10 seconds. The absorption of ASL and the fluid meniscus was measured from the weight change of the filter paper. The weight of the filter paper was measured using an analytical balance (Sartorius BP61S; Sartorius AG, Göttingen, Germany).

[0550] Example 4.4: Desensitization and Challenge with OVA Experiments were conducted using 8-week-old BALB / c mice. The experimental protocol was approved by the Yonsei University Animal Ethics Committee. Mice were housed and maintained in an animal facility under standard laboratory conditions [12 / 12 hour light / dark cycle, controlled temperature (21 ± 2 °C) and humidity (55 ± 5%) and access to food and water ad libitum]. All experiments were performed in accordance with the guidelines of the Yonsei University Animal Research Committee. Mice were divided into three groups, a vehicle group (PBS-primed and challenged mice treated with vehicle), an OVA group (OVA-primed and challenged mice treated with vehicle) and an OVA+F56 group (OVA-primed and challenged mice treated with F56). 8-week-old mice of appropriate age and sex were primed on days 0 and 14 by i.p. injection of OVA (Sigma-Aldrich). The priming emulsion consisted of 50 μg of OVA and 2 mg of aluminum potassium sulfate in 200 μL of saline. On days 21, 22 and 23, the primed mice were lightly anesthetized by inhalation of isoflurane and challenged with 100 μg of OVA in 30 μL of saline administered intranasally. Control group mice were treated in the same manner as PBS. F56 (10 mg / kg) was administered by i.p. injection 12 hours prior to i.n. OVA challenge, respectively.

[0551] Example 4.5: Evaluation of Airway Reactivity to Methacholine Challenge Airway responsiveness to methacholine (MCh) was measured 24 hours after the last OVA exposure using the FlexiVent system (Scireq, Montreal, QC, Canada). Mice were anesthetized by intraperitoneal injection of a mixture of Zoletil (30 mg / kg; Virbac Laboratories, Carros, France) and Rompun (10 mg / kg; Bayer, Leverkusen, Germany), then tracheotomized and connected to the FlexiVent. Baseline airway resistance was measured after saline spraying for 10 seconds using an Aeroneb ultrasonic nebulizer (SCIREQ). After baseline measurement, the mice were exposed to increasing concentrations of nebulized methacholine (0, 1.56, 3.13, 6.25, 12.5, and 25 mg / mL). Also, bronchial airway responsiveness was measured using the whole body plethysmography method (Buxco, Miami, FL, USA). Enhanced pause (Penh) was used as the main index of airway responsiveness. Penh was measured for 2 minutes under baseline conditions. Then, the mice were exposed to inhalation of PBS or MCh (25 mg / mL) for 2 minutes. The results of Penh are presented as absolute values.

[0552] Example 4.6: Bronchoalveolar Lavage Bronchoalveolar lavage (BAL) was performed after evaluation of airway responsiveness. Cells were pelleted by centrifugation and resuspended in PBS to obtain cell counts. Cytospin was prepared using a cytocentrifuge (Shandon Cytospin4 cytocentrifuge; Thermo Scientific, MA, USA, USA) and stained with a Diff-Quik Stain Set (Dade Behring, USA, DE, USA) to evaluate inflammation.

[0553] Example 4.7: Histological Evaluation of Inflammation For the evaluation of airway inflammation, the left lung was fixed in 4% paraformaldehyde overnight and embedded in paraffin. Histological slides were prepared from 5-μm sections and stained with hematoxylin and eosin (H&E). The inflammation score indicating the severity of bronchial inflammation was evaluated by three blinded observers using a previously reported method. To evaluate goblet cell hyperplasia, the sections were stained with periodic acid-Schiff (PAS) using a PAS staining kit (Sigma-Aldrich) according to the manufacturer's protocol. Nasal mucosa and HNE cells harvested on Transwell were gently washed with PBS and fixed in 4% paraformaldehyde. After deparaffinization and hydration, the slides were immersed in periodic acid solution at room temperature for 5 minutes. After washing off with distilled water, the slides were immersed in Schiff's reagent at room temperature for 15 minutes and then washed with running tap water for 5 minutes.

[0554] Example 4.8: Measurement of OVA-Specific IgE OVA-specific IgE in serum was measured using an Anti-Ovalbumin IgE (mouse) ELISA kit (Cayman Chemical, Ann Arbor, MI, USA) according to the manufacturer's protocol. To measure the level of OVA-specific IgE in serum, diluted serum was added to anti-IgE antibody-precoated 96-well plates and then cultured with OVA-biotin conjugate. The bound biotinylated OVA was detected with streptavidin-horseradish peroxidase (HRP) using 3,3',5,5'-tetramethylbenzidine (TMB) as a substrate.

[0555] Example 4.9: Epithelial SCN- Transport Nasal epithelial cells were plated on Transwell permeable supports and cultured at the air-liquid interface (ALI) for 14 days. After culturing fully differentiated epithelial cells with IL-4 (Invitrogen, 10 ng / mL) for 48 hours, F56 (30 μM) or vehicle was added for 30 minutes. The Transwell inserts with cells were washed with PBS, and the basal side was incubated with 10 mM glucose and 5 μCi of S14 CN (total SCN concentration - : 86 μM) and treated with 1 mL of PBS. The top side of the Transwell was filled with 0.5 mL of PBS containing 5 μM amiloride to block epithelial sodium. Next, the fluid at the top was collected every 5 minutes and placed in scintillation vials for radioactivity assessment.

[0556] Example 4.10: Measurement of T3 and T4 Female balb / c mice were administered i.p. with 10 mg / kg of F56 every 24 hours over a 7-day period to test the negative effects of F56. Twenty-four hours after the last administration, the hearing of the mice was measured and plasma was obtained. According to the manufacturer's protocol, the total plasma levels of triiodothyronine (T3) or thyroxine (T4) were analyzed using a mouse ELISA kit (Calbiotech, T3043T-100 or T4044T-100).

[0557] Example 4.11: Auditory Brainstem Response (ABR) The auditory level in each mouse was measured by measuring the ABR threshold using an auditory-evoked potential workstation and BioSig software (Tucker-Davis Technologies, Alachua, FL, USA). The output from the speaker was calibrated using a PCB 377C10 microphone (PCB Piezotronics, Inc., New York, NY, USA) and was found to be within ±4 dB for the tested frequency range. After anesthetizing the mice, each ear was stimulated with an ear probe sealed inside the ear canal. The body temperature of the mice was maintained at 38 °C with an isothermal heating water pad. The intensity of the click sound decreased from 70 dB SPL to 10 dB SPL in 5-dB decrements. The average value of the ABR was calculated, and the auditory threshold was defined as the lowest recognizable ABR response until wave I of the ABR could no longer be visually distinguished.

[0558] Example 4.12: In Vitro Optical Imaging Eight-week-old NF-β luciferase-dTomato reporter mice (Korean Mouse Phenotype Center) were sensitized on days 0 and 14 by i.p. injection of OVA (50 μg, Sigma-Aldrich) and 2 mg of aluminum potassium sulfate. On days 21, 22, and 23, the sensitized mice were anesthetized by inhalation of isoflurane and challenged with 150 μg of OVA in 30 μL of saline administered intranasally. Control group mice were treated in the same manner with PBS. F56 (10 mg / kg) was administered i.p. 12 h before i.n. respectively. The NF-κB expression in the lungs of NF-kB reporter mice was compared using an IVIS system. Lungs were isolated from the mice and fluorescence imaged using an IVIS spectrum (Ex570, Em620). The average fluorescence density of the images was analyzed using Living Image4.3.1 software.

[0559] Example 4.13: Measurement of Tracheal Contraction All experiments were performed using 4-week-old male Sprague-Dawley rats. The experimental protocol was approved by the Yonsei University Animal Ethics Committee. Rats were sacrificed and tracheal strips were isolated. After separating the connective tissue, the trachea was cut into 3-mm-long rings. The trachea was mounted under a tension of 1 g for tension recording and equilibrated for 1 h in an organ bath containing 25 mL of oxygenated physiological solution. The solution was continuously gas-treated with 95% O 2 and 5% CO 2 At 1 h after stabilization, a sustained contraction response was induced in the organ bath using 0.3 μM carbachol (CCh). When a sustained tension was established, F56 (30 μM), forskolin (10 μM), and IBMX (100 μM) were applied to the bath. Tracheal contractions were measured using a FORT 10G transducer (World Precision Instruments, Sarasota, FL, USA) and a PowerLab4 / 35 (AD Instruments, Castle Hill, Australia). Data were recorded and analyzed using Labchart Pro7 (AD Instruments). Pendrin (SCL26A4), a membrane - spanning anion exchanger, exchanges Cl - for HCO 3 - , I - , OH - and SCN - with bases such as these, and is the most highly up - regulated gene in bronchial biopsies from asthmatic patients. Interestingly, patients with pendrin mutants have a low asthma incidence, and pendrin - null mice show reduced allergic airway inflammation. The inventors isolated a novel pendrin inhibitor, F56 (2-(4-(tert - butyl)phenyl)-4-(thiophen - 2 - ylmethylene)oxazol - 5(4H)-one), which has a potent therapeutic effect on allergic inflammation in a mouse model of ovalbumin (OVA) - induced asthma.

[0560] F56 was identified from 54,400 synthetic compounds by cell - based high - throughput screening (Figure 5(a), A). F56 potently inhibited pendrin - mediated Cl - / SCN - , Cl - / I - , Cl - / HCO 3 - and Cl - / OH - exchange activities in a dose - dependent manner (Figure 5(a), B and C, and Figure 6(a), A - C). F56 showed no cytotoxicity up to 30 μM in NIH3T3 and CHO - K1 cells and more potently inhibited pendrin activity than the recently identified pendrin inhibitors, PDS inh -A01 and PDS inh -C01 (Figure 6(a), D - I). On the other hand, F10 was confirmed to show no cytotoxicity up to 30 μM in NIH3T3 and CHO - K1 cells like F56 (Figure 6(b)). F56 inhibited human and mouse pendrin - mediated Cl - / I - exchange with almost the same efficacy (Figure 7(a), A and B). F56 had an IC 50 >100 μM for Cl- / HCO 3 - Weakly suppressed the exchange activity and had no effect on SCL26A7 and SLC26A9. Cystic fibrosis transmembrane conductance regulator (CFTR), anoctamin-1 (ANO1), human ether-a-go-go-related gene (hERG) channel and 5-HT 2A activity was not affected by F56 (Figure 7(a), C-J). On the other hand, F10 was confirmed to have no effect on the activities of CFTR, ANO1 and

[0561] the hERG ion channel (Figure 7(b)). In the primary cultures of human nasal epithelium (HNE) and human bronchial epithelium (HBE) cells, IL-4 treatment strongly upregulated pendrin expression and pendrin-mediated Cl - / HCO 3 - exchange activity, and F56 potentially suppressed pendrin-mediated Cl - / HCO 3 - exchange activity (Figure 5(a), D-F and Figure 8(a), A-C). The present inventors observed the long-term treatment effect of F56 on the functional expression of pendrin and other ion channels involved in ASL regulation. Interestingly, the IL-4-induced upregulation of Cl - / HCO 3 - exchange activity and the protein expression level of pendrin were strongly decreased by long-term treatment with F56 without a change in the mRNA expression level, while the IL-4-induced upregulation of ANO1 was not affected (Figure 5(a), G-I). On the other hand, F10 was confirmed to have no effect on the mRNA expression level of PDS like F56 (Figure 5(b)). The mRNA expression levels and ion channel activities of ANO1, CFTR and ENaC were not altered by long-term treatment with F56 (Figure 8(a), D-F). On the other hand, F10 was confirmed to have no effect on the mRNA expression levels of ANO1, CFTR and ENaC like F56 (Figure 8(b)).

[0562] Pretreatment with F56 significantly attenuated OVA-induced airway hyperresponsiveness (Fig. 9(a), A), and decreased the numbers of eosinophils and neutrophils in the BALF of OVA-challenged mice. As a result of histological analysis, increased inflammatory infiltration, epithelial thickness, and the increased number of goblet cells in the central airways were attenuated by treatment with F56 in OVA-challenged mice. The mean inflammation score of OVA-challenged mice was significantly lower in the F56-treated group than in the untreated group, but the serum level of OVA-specific IgE was not altered by F56, indicating that F56 does not act on the general allergic reaction mechanism (Fig. 10). On the other hand, F10 was confirmed to decrease the increased airway resistance in an OVA-induced asthmatic mouse model like F56 (Fig. 9(b)).

[0563] According to recent studies, it has been revealed that the activation of NF-kB by the increased production of hypothiocyanite (OSCN - ) due to the upregulation of pendrin, peroxidases, and dual oxidase (Duox1 / Duox2) in airway epithelium is involved in allergic airway inflammation. The concentration of SCN - at the apical surface was significantly increased by IL-4 treatment and suppressed by F56 (Fig. 9(a), B). Real-time PCR analysis showed that IL-4 treatment significantly increased the mRNA expression level of Duox1 but did not affect Duox2, and F56 did not affect the mRNA expression levels of Duox1 or Duox2 (Fig. 9(a), C). Pretreatment with F56 significantly suppressed the IL-4-induced activation of NF-kB in HNE cells (Fig. 9(a), D). In particular, intranasal administration of NaSCN significantly blocked the inhibitory effect of F56 on airway hyperresponsiveness and abolished the protective effect of F56 on lung injury in OVA-challenged asthmatic mice (Fig. 9(a), E and F). Also, F56 significantly blocked the activation of NF-kB in the lungs of OVA-treated mice, and SCN -The treatment of [[ID=]] suppressed the protective effect of F56 in transgenic NF-κB reporter mice (Figure 10, G). Application of F56 to an established model of allergic asthma significantly decreased OVA-induced airway hyperresponsiveness and PAS-positive cells in the airway epithelium (Figure 11). Such results suggest that F56 is useful for the prevention and treatment of allergic asthma.

[0564] Pendrin is associated with the airway inflammation-mediated upregulation of MUC5AC in the airway epithelium. In particular, according to the present invention, it was shown that treatment with F56 significantly decreased the mRNA expression level of IL-4-induced MUC5AC (Figure 12(a), A). Also, pretreatment with F56 decreased IL-4- and IL-13-induced goblet cell hyperplasia in HNE cells (Figure 12(a), B). The thickness of the ASL in primary cultures of airway epithelial cells treated with IL-13 was significantly higher in pendrin-null mice with a mutant SLC26A4 gene and in hearing-impaired patients compared to the control group. In HNE cells expressing wild-type pendrin, IL-4 and IL-13 treatment strongly increased the protein expression level of pendrin compared to the control group, decreased the total volume of the ASL and the fluid meniscus, and F56 treatment hardly decreased the total volume of the ASL and the fluid meniscus. However, the total volume was not altered by IL-4 and F56 in HNE cells expressing mutant pendrin (Figure 12(a), C-F). Such results are consistent with the previous research findings that inh -A01 significantly increased the depth of the ASL in IL-13-treated HBE cells. The suppression of MUC5AC and the increase in the thickness of the ASL by F56 can provide additional beneficial effects in airway inflammatory diseases such as COPD, cystic fibrosis, and asthma. On the other hand, it was confirmed that F10 decreased the mRNA expression level of MUC5AC in a concentration-dependent manner like F56 (Figure 12(b)).

[0565] Since patients with SLC26A4 gene mutations are associated with prelingual deafness and goiter, changes in auditory or thyroid hormone levels can be induced by pendrin inhibition. However, after 1 week of treatment with F56 (10 mg / kg / day), the auditory thresholds and plasma levels of T3 and T4 did not change (Figure 13, A - C). Also, F56 did not affect the contraction of tracheal smooth muscle (Figure 13, D).

[0566] In summary, F56 reduced airway hyperresponsiveness and airway inflammation by suppressing the SCN - / NF - kB pathway. F56 also showed a protective effect against IL - 4 - and IL - 13 - induced goblet cell hyperplasia and ASL deficiency (Figure 14). These results indicate that pendrin inhibitors are promising candidates for the treatment of allergic asthma.

[0567] Example 5: Function of New Compound (F56) in Reducing Lipopolysaccharide-Induced Acute Lung Injury in Mice Example 5.1: Experimental Animals Wild-type male C57BL / 6J mice, 8 - 10 weeks old and weighing 20 - 24 g, were purchased from Orient Bio (Sungnam, Republic of Korea). All animals were supplied with food and water and subjected to a similar diurnal light cycle. Transgenic NF-κ reporter / SPC-Cre-ERT2 mice were used in the present invention for IVIS. Briefly, the NF-κ reporter mice contain a ROSA26 lox-STOP-lox cassette inserted between the promoter and the NF-κ gene. Generally, the stop gene is located between loxP and loxP, and NF-κ is not expressed. ROSA26R mice were bred with surfactant protein C (SPC)-Cre-ERT2 mice to obtain NF-κ reporter / SPC-Cre-ERT2 mice. The activity of the Cre-ERT2 recombinase in these transgenic mice was induced by tamoxifen. These NF-κ reporter / SPC-Cre-ERT2 mice express NF-κ activity in alveolar epithelium through either dTomato fluorescence or luciferase in the presence of tamoxifen. Tamoxifen (Sigma, USA) was dissolved in a 10:1 mixture of sunflower seed oil / ethanol (10 mg / mL). Each 4-week-old mouse was intraperitoneally injected with 100 μL of tamoxifen / day for 5 consecutive days. One week after the last injection, the mice were used for IVIS or in vivo imaging. Transgenic NF-κ reporter and SPC-Cre-ERT2 mice were received from Yonsei University.

[0568] Example 5.2: LPS-Induced ALI in a Mouse Model Mice were lightly anesthetized by isoflurane inhalation (Abbott Laboratory) and maintained in a supine position while raising the head. LPS (Escherichia coli, O111:B4, Sigma) (10 mg / kg) in 50 μL PBS was administered by intranasal (i.n.) inhalation. The control group received 50 μL of sterile PBS intranasally. With the assistance of a microsyringe from Hamilton, the administration solution was gradually released into the nostrils. In the pretreatment model, F56 (10 mg / kg) in 50 μL DMSO was administered intraperitoneally (i.p.) 1 hour before LPS inhalation. In the post-treatment model, two doses of F56 were administered at 6 and 12 hours after LPS inhalation. Mice in the pretreatment group were euthanized, and the lungs were harvested 48 hours after LPS inhalation. In the post-treatment group, euthanasia and sample collection occurred 24 hours after LPS administration. SCN - For the experiment, after F56 treatment, 50 μL of NaOH, NaHCO 3 , or NaSCN (100 mM) was administered intranasally. PBS was administered to the control group in the same manner.

[0569] Example 5.3: Isolation of Bronchoalveolar Lavage All mice were euthanized with a lethal overdose of ketamine and xylazine. BALF was obtained by tracheal cannulation with 1 mL of sterile saline. The BALF was centrifuged (4 °C, 3000 rpm, 10 min), and the supernatant was stored at -80 °C for additional analysis. The cell pellet was resuspended in 100 μL of PBS and used for cell counting and cytospin samples. The total cell count of each sample was determined using a hemocytometer (Marienfield) according to the manufacturer's protocol. After transferring 90 μL aliquots of each sample into a slide chamber, the slides were inserted into the cytospin with the outside facing up. After centrifuging the slides at 800 rpm for 5 min, the slides were removed from the cytocentrifuge and allowed to dry before staining. Cytospins were prepared using a cytocentrifuge (Shandon Cytospin 4 cytocentrifuge, Thermo Scientific, Waltham, MA, USA) and stained with a Diff-Quik Stain Set (Dade Behring, Newark, DE, USA) to evaluate inflammation. The protein concentration of the BAL supernatant was measured using a BCA assay (Thermo Fischer Scientific). 2 μL aliquots of each sample and 198 μL of working reagent were pipetted into a microplate well and mixed thoroughly on a plate shaker for 30 min. After incubating at 37 °C for 30 min, the plate was cooled and the absorbance at 562 nm was read on a spectrophotometer.

[0570] Example 5.4: Harvesting of Lung Tissue and Histological Examination The right lung was isolated, the pulmonary vasculature was flushed with saline under low pressure, and then stored at -80 °C prior to protein extraction. Until the pleural margin became sharp, 25 cm H 2The left lung was inflated by tracheotomy with low melting point agarose (4%) in PBS at the pressure of O. Then, the lung was excised, fixed overnight with 10% formaldehyde in PBS, and embedded in paraffin in 5-μm sections. The left lung portion was stained with H&E and evaluated subjectively by light microscopy. Histopathology was examined in a blinded fashion by two qualified researchers. Five pathologic processes that could be easily identified were scored using the weighted scale presented in the official ATS workshop report. Lung sections were processed for immunohistochemistry using anti-rabbit SLC26A4 (ab98091, abcam) antibody.

[0571] Example 5.5: Cl- / SCN- Exchange Measurement Human pendrin (PDS) and YFP-F46L / H148Q / I152L transiently transfected human alveolar epithelial cells (hAEC) were plated in 96-well plates at a density of 2 × 10 4 cells per well and cultured for 48 hours. Each well of the 96-well plate was washed twice with 200 μL of PBS and filled with 100 μL of PBS. To measure the effect of F56 on hPDS-mediated Cl - / SCN - exchange activity, the cells were pretreated with F56. After culturing at 37 °C for 10 minutes, the 96-well plate was placed on a reverse fluorescence microscope (Nikon, Tokyo, Japan) equipped with a cooled charge-coupled device camera (Zyla sCMOS), and image acquisition and analysis software (Meta Imaging Series 7.7). Each well was individually analyzed for hPDS-mediated SCN - influx by continuously recording YFP fluorescence for 4 seconds (2 seconds per point). Then, 100 μL of 140 mM SCN - solution was added in 4 seconds, and YFP fluorescence was recorded for 14 seconds.

[0572] Example 5.6: Real-Time RT-PCR Analysis Total messenger RNA (mRNA) was extracted using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) and reverse-transcribed using random hexamer primers, oligo(dT) primers, and SuperScript® III Reverse Transcriptase (Invitrogen). Quantitative real-time PCR was performed using a StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) and Thunderbird SYBR qPCR mix (Tokyobo, Osaka, Japan). The thermal cycling conditions included an initial step of 95°C for 5 minutes in a 96-well reaction plate, followed by 40 cycles of 95°C for 10 seconds, 55°C for 20 seconds, and 72°C for 10 seconds. The primer sequences were listed in Table 3 above.

[0573] Example 5.7: ELISA Levels of Macrophage inflammatory protein (MIP-2), interleukin-1β (IL-1β), and tumor necrosis factor α (TNF-α) in lung lysates were measured using an ELISA kit (Millipore) according to the manufacturer's instructions.

[0574] Example 5.8: Measurement of Protein Expression Level Lung tissues were harvested and homogenized in lysis buffer (PRO-PREP TMThe extracted solution was dissolved in iNtRON Biotechnology. The samples were centrifuged at 13,000 g for 30 minutes at 4°C. The protein concentration of the supernatant was determined by BCA analysis (Thermo Fisher Scientific). The same amount of protein was separated by SDS / PAGE and transferred to a nitrocellulose membrane. The membrane was blocked with 5% skim milk in TBS-T (TBS (170-6435, Bio-Rad Laboratories) and 1% Tween-20 (170-6531, Bio-Rad Laboratories)) for 1 hour at room temperature. Subsequently, the membrane was incubated overnight with the primary antibody diluted with 5% skim milk in TBS-T at 4°C. After washing with TBS-T, the blot was incubated with horseradish peroxidase-conjugated secondary antibody and 5% skim milk in TBS-T for 1 hour at room temperature and then developed using the Super-Signal West Pico chemiluminescence detection kit. The antibodies used in the present invention included rabbit SLC26A4 (ab98091, abcam), mouse phospho-Iκ (9246, Cell Signaling Technology), mouse Iκ (4814, Cell Signaling Technology), and rabbit α-tubulin (PA5-16891, Cell Signaling Technology). Western blot quantification was performed using ImageJ (Image Processing and Analysis in Java; NIH, USA) software.

[0575] Example 5.9: IVIS Imaging of live animals and tracheas was performed using an IVIS kinetic imaging system (Caliper Life Sciences, Preston Brook Runcorn, UK). The IVIS system consisted of a cooled charge-coupled device camera mounted in a light sample chamber. Fluorescent excitation light was provided by a halogen lamp combined with an appropriate excitation filter. A emission filter was placed in front of the camera aperture so that light rays of a specific wavelength could be recorded according to the emission spectrum of the irradiated FP. Fluorescent imaging was obtained at an excitation wavelength of 554 nm and an emission wavelength of 581 nm (dTomato). Mice were divided into three groups for IVIS imaging (DMSO + PBS, DMSO + LPS and F56 (10 mg / kg, i.p) + LPS), and ex vivo lungs were aseptically removed 6 hours after LPS treatment. When imaging the organs, they were placed as flat as possible to allow for complete and consistent light penetration to minimize potential variations in measurements due to differences in tissue thickness. Fluorescence was quantified using the region of interest tool in Living Image software (version 3.2, Caliper Life Sciences).

[0576] Example 5.10: Collection of Human Bronchoalveolar Lavage Fluid Forty-one patients with ARDS due to pneumonia who had undergone bronchoalveolar lavage (BAL) were classified into the ARDS group. Twenty-five patients who were admitted for SPN evaluation without evidence of lung infection were classified as the control group at Severance Hospital between May 2013 and September 2015. Before bronchoscopy, the subjects received topical anesthesia (lidocaine) by nebulizer and were then sedated with midazolam and pentanyl. The bronchoscope was inserted and wedged in the mouth for BAL. BAL was performed according to a standardized protocol (pneumonia group: bronchi of lung lesions, control group: contralateral bronchi from lung mass), and 10 cc of BALF was obtained from each patient using approximately 30 mL of 0.9% sterile saline. The BALF was centrifuged (10 minutes; 1500 g), and the supernatant was cryopreserved at -80 °C until use. Demographic and clinical data, including age, gender, body mass index (BMI), comorbidities, BALF analysis, cause of pneumonia, and final diagnosis, were obtained from both medical records and each participant. The pendrin level in the supernatant was measured using a human SLC26A4 ELISA kit (MBS764789, Mybiosource) according to the manufacturer's instructions.

[0577] Example 5.11: Research Approval All animal protocols were approved by the Institutional Animal Care and Use Committee of Yonsei University College of Medicine (2016-0322). All animal experiments were performed in accordance with the recommendations of the Guidelines for the Care and Use of Laboratory Animals of the National Institutes of Health. The human research protocol was reviewed and approved by the Institutional Review Board of Severance Hospital, Yonsei University Health System in Seoul, Korea (ARDS group IRB No. 4-2013-0585, control group IRB No. 4-2014-1014). Written informed consent was obtained from the patients or their guardians regarding the use of BALF samples.

[0578] As expected, intratracheal injection of LPS induced ALI in WT mice. Total cell counts and protein concentrations in bronchoalveolar lavage fluid (BALF) increased significantly after LPS treatment (Figure 15A). Lung histology also showed leukocyte infiltration and lung injury in WT mice (Figure 15B). In contrast, LPS did not increase cell counts or protein concentrations in pendrin-null mice (Figure 15C). Furthermore, lung histology showed a lack of leukocyte infiltration and lung injury in pendrin-null mice after LPS treatment (Figure 15D). The mean body weight change after 48 hours of LPS injection was more pronounced in WT mice compared to pendrin-null mice (-3.38 g vs. -1.75 g, p < 0.01, Figure 1E). Immunoblot analysis of lung tissue showed an increase in pendrin protein expression after LPS treatment compared to the vehicle control group (Figure 15F). These results suggested that pendrin plays an essential role in the development of LPS-induced ALI.

[0579] As described above, for the novel pendrin inhibitor, F56, 54,400 synthetic compounds were identified by high-throughput screening. F56 strongly inhibited Cl - / I - 、Cl - / SCN - 、Cl - / HCO 3 - and Cl - / OH - exchange activities and strongly decreased the protein expression level of pendrin in nasal tracheal epithelium, but did not decrease the mRNA expression level of pendrin.

[0580] We investigated the effects of LPS and F56 on the mRNA and protein levels of pendrin in human alveolar epithelial cells (hAEC). LPS treatment significantly increased the mRNA and protein expression levels of pendrin, and F56 decreased the protein expression level of pendrin, but did not alter the mRNA expression level of pendrin in hAEC (Figure 16B and Figure 16C). Also, F56 inhibited the Cl- / SCN - Exchange activity (IC 50 = 4.7 ± 0.82 mM) was strongly inhibited (Figure 16D). As described above, upregulation of pendrin can increase the production of hypothiocyanite (OSCN - ) by upregulating dual oxidase (Duox1 / Duox2) in the airway epithelium of allergic inflammation and activate NF-β. Real-time PCR analysis showed that LPS treatment significantly increased the mRNA expression level of Duox2, and F56 did not change the mRNA expression level of Duox2 (Figure 16E).

[0581] To examine the protective function of F56 in the LPS-induced ALI mouse model, mice were treated with F56 1 hour before intranasal injection of LPS and euthanized 48 hours after LPS administration (Figure 17A). It was revealed that mice pretreated with F56 (10 mg / kg) had decreased total cell numbers and protein concentration levels in BALF compared to vehicle-treated mice (Figure 17B and Figure 17C). In addition, F56 pretreatment significantly decreased the lung injury score compared to vehicle-treated mice with leukocyte infiltration after LPS exposure was suppressed (Figure 17D and Figure 17E). To determine whether F56 treatment was effective after LPS injury, mice were treated with F56 6 hours and 12 hours after intranasal injection of LPS and then euthanized 24 hours after LPS administration (Figure 17F). After LPS injection, F56 treatment significantly decreased not only the lung injury score but also the total cell numbers and protein concentration in BALF, consistent with the results of the F56 pretreatment experiment (Figure 17G - Figure 17I).

[0582] To confirm the underlying therapeutic effect mechanism of F56 in LPS-induced ALI, the inventors examined the anions (OH - , HCO 3 - , and SCN -) was administered. Intranasal application of NaSCN (50 μL of 100 μM) blocked the protective effect of F56 in LPS-induced ALI; the total cell count and lung injury score of BAPS increased compared to the group treated with LPS alone. However, administration of NaOH and NaHCO 3 did not change the effect of F56 in LPS-induced ALI mice (Figures 18A and 18B). Also, histological analysis revealed that the protective effect of F56 against inflammatory cell infiltration and lung injury was abolished by NaSCN administration after LPS administration (Figure 18C). More interestingly, co-application of LPS and NaSCN induced severe lung injury in pendrin-null mice, whereas administration of LPS alone did not induce ALI (Figure 18D). These data strongly suggest that the therapeutic effect of F56 results from the inhibition of the SCN - transport function of pendrin.

[0583] The inventors further dissected the signaling pathway by the action of F56 in a LPS-induced ALI model using NF-κ reporter / SPC-Cre-ER T2 mice. Quantitative fluorescence was measured using in vivo optical imaging (IVIS) images of mouse lungs that were aseptically removed immediately before imaging. The fluorescence of the excised lungs increased after LPS treatment, which was inhibited by F56 (Figures 19A and 19B). Such results suggested that the activation of NF-κ was inhibited by F56 in LPS-injected mice. Immunoblot analysis showed that the NF-κ pathway was involved in the action of F56. The expression of phospho-Iκ protein, indicating the activation of NF-κ, increased after LPS administration, and pretreatment with F56 before LPS significantly decreased the phospho-Iκ expression (Figures 19C and 19D).

[0584] The levels of cytokines including IL-1β, tumor necrosis factor-α, and macrophage inflammatory protein (MIP)-2 were significantly increased after LPS administration compared to PBS (Figure 19E-19H). Although the difference was not statistically significant, IL-6 tended to increase compared to PBS. In contrast, the levels of pro-inflammatory cytokines were decreased in F56-pretreated mice compared to those treated with vehicle (DMSO) after LPS administration (Figure 19E-Figure 19H).

[0585] To translate in vitro and in vivo findings to human disease, the inventors measured patients with pneumonia (ARDS group, n = 41) and patients with influenza pneumonitis nodules (SPN) but without infection (control group, n = 25). The clinical characteristics of the patients are shown in Table 6 below.

[0586]

Table 6

[0587] There was no significant difference in mean age between the control group and the ARDS group (63.8 vs. 65.9, p = 0.517), and males were dominant in both groups (80% vs. 78%, p = 0.851). The average length of hospital stay among ARDS patients was 36 days, and the 28-day mortality rate was 24.4% (Table 1). Pendrin levels were significantly increased in the BALF of ARDS patients (n = 41) compared to the control group (n = 25) (mean, 24.86 vs. 6.83 ng / mL, p < 0.001) (Figure 20).

[0588] Overall, the new evidence strongly suggests that pendrin is a core protein in the development of airway inflammatory diseases including asthma, chronic obstructive pulmonary disease and rhinitis. The inventors have demonstrated that the expression level of pendrin increased in the airway of LPS-treated mice. In addition, the inventors have not developed in LPS-induced ALI pendrin null mice, which strongly represents an important role of pendrin in the etiology of ALI. This is consistent with recent reports that the expression of pendrin is improved in LPS-induced ALI and that non-specific pendrin inhibitors attenuated ALI in mice. Such evidence encouraged the inventors to develop pendrin inhibitors as new drugs for the treatment of ALI. The inventors screened 54,400 synthetic compounds and discovered a specific pendrin inhibitor (F56) that does not affect other ion transports such as cystic fibrosis transmembrane conductance regulator (CFTR) and calcium-activated chloride channel (CaCC). The expression of pendrin was upregulated by LPS treatment in human alveolar epithelium, which was effectively suppressed by F56. Surprisingly, F56 almost completely blocked the development of LPS-induced ALI in mice. In addition, administration of F56 attenuated lung injury in mice after LPS treatment, indicating that the clinical treatment period of pendrin inhibitors is wide enough to include the period after ALI. The inventors have shown increased expression of pendrin in BALF from pneumonia patients and in the airway of LPS-treated mice, which strongly suggests a high potential for the clinical application of pendrin inhibitors in inflammatory airway diseases.

[0589] The basic mechanisms of the role of pendrin and the therapeutic effect of F56 in the ALI model are not clear. The inventors focused on the Cl - / SCN - exchange activity of pendrin and hypothiocyanite (OSCN - ), which is synthesized from SCN - transported through various anion transporters (including pendrin) by lactoperoxidase in the airway epithelium. OSCN -is known to be part of an important innate defense system against microorganisms in the airway and induces airway inflammation in airway epithelium. According to recent studies, IL-4 upregulates the Cl - / SCN - exchange activity, increases the production of OSCN - , which induces the activation of NF-κ and induces airway inflammation in a murine allergic asthma model. The inventors have shown that when NaSCN is added to the airways of mice, the therapeutic effect of F56 against lung injury is abolished. Also, the application of NaSCN induced lung injury even in pendrin-null mice, but LPS-induced ALI did not occur. Such data indicate that airway surface SCN - transported by pendrin is an essential component of LPS-induced airway inflammation.

[0590] NF-κ is an important determinant of the inflammatory response in the airway, and its inhibition attenuates in vivo ALI. Also, the inventors have observed that F56 suppresses the activation of LPS-induced NF-κ and subsequent cytokine production in a murine ALI model and in alveolar epithelium. Collectively, the data according to the present invention suggest that the mode of action of the pendrin inhibitor blocks the transepithelial transport of SCN - , and then OSCN -It has been shown to result from F56, which suppresses the production of [[ID=]] and the activation of NF-κ. This suppressed the production of pro-inflammatory cytokines (Figure 21). This is a mode of action very similar to that discovered in a murine asthma model in which a pendrin inhibitor attenuates OVA-induced allergic airway inflammation by suppressing the pendrin / OSCN− / NF-κ cascade (18). However, considering previous reports (28, 29) showing that LPS can activate NF-κ through the TLR4 / MyD88 pathway, the reason why YS-01 almost completely suppresses LPS-induced ALI is unknown. However, the lack of the ALI phenotype induced by NaSCN in pendrin-null mice strongly suggests that pendrin-mediated OSCN predominantly activates the NF-κ cascade in the LPS-induced ALI model. This is a mode of action very similar to that discovered in the murine asthma model according to the present invention in which a pendrin inhibitor attenuates OVA-induced allergic airway inflammation by suppressing the pendrin / OSCN− / NF-κ cascade. However, considering previous reports showing that LPS can activate NF-κ through the TLR4 / MyD88 pathway, the reason why F56 almost completely suppresses LPS-induced ALI is unknown. However, the lack of the ALI phenotype induced by NaSCN in pendrin-null mice strongly suggests that pendrin-mediated OSCN− predominantly activates the NF-κ cascade in the LPS-induced ALI model.

[0591] Important treatments for ALI patients have improved, but the ALI / ARDS mortality rate remains high and the medical treatment options for ALI / ARDS are limited. Since F56 showed a strong therapeutic effect in the murine ALI model, pendrin may be a new target for the treatment of ALI / ARDS. The upregulation of pendrin expression in the BALF of pneumonia patients raises the hope of increasing the potential clinical therapeutic benefits of pendrin inhibitors for ALI / ARDS. In addition, F56 is low in cytotoxicity, chemically stable, and operates at the nanomolar level; it is an excellent compound for further development as a final candidate for clinical trials. In other words, the inventors have demonstrated that pendrin is essential for LPS-induced ALI and that a compound (F56) that inhibits pendrin strongly suppresses LPS-induced ALI. According to the present invention, pendrin inhibitors are of a promising new drug class for the treatment of ALI.

Claims

1. A compound selected from the group consisting of the following compounds described in Table 1, its E-isomer or Z-isomer, its optical isomer, a mixture of two isomers among the E-isomer, the Z-isomer and the optical isomer, its pharmaceutically acceptable salt or its solvate. 【Table 1】

2. A pharmaceutical composition for preventing or treating respiratory diseases, comprising as an active ingredient the compound according to Claim 1, its E-isomer or Z-isomer, its optical isomer, a mixture of two isomers among the E-isomer, the Z-isomer and the optical isomer, its pharmaceutically acceptable salt or its solvate.

3. The pharmaceutical composition according to Claim 2, wherein the respiratory disease is an inflammatory airway disease.

4. The inflammatory airway disease is one or more selected from the group consisting of asthma, acute or chronic bronchitis, allergic rhinitis, acute respiratory infections, acute upper respiratory tract infections, cystic fibrosis, acute respiratory distress syndrome (ARDS), acute lung injury (ALI) and chronic obstructive pulmonary disease (COPD). The pharmaceutical composition according to Claim 3.

5. The pharmaceutical composition according to Claim 2, wherein the active ingredient acts as a pendrin inhibitor.

6. The pharmaceutical composition according to Claim 2, wherein the active ingredient specifically controls channels associated with respiratory diseases.

7. The pharmaceutical composition according to Claim 2, wherein the active ingredient preserves the volume of airway surface liquid (ASL) and reduces the separation of mucin.

8. The pharmaceutical composition according to Claim 2, further comprising a pharmaceutically acceptable carrier.

9. The pharmaceutical composition according to Claim 2, further comprising other pharmaceutical components.

10. A health functional food comprising as an active ingredient the compound according to Claim 1, its E-isomer or Z-isomer, its optical isomer, a mixture of two isomers among the E-isomer, the Z-isomer and the optical isomer, its pharmaceutically acceptable salt or its solvate.

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