New compounds and compositions comprising the same as active ingredients for the prevention or treatment of respiratory diseases

Abstract

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

Description

Technical Field

[0001] This invention relates to a novel compound and compositions comprising the compound as an active ingredient for the prevention or treatment of respiratory diseases. Background Technology

[0002] Pendrin, encoded by the SLC26A4 gene, is an anion exchanger and a member of the SLC26 gene family. It exchanges Cl- - Exchanged into HCO3 - I - OH - and SCN - Panterin is a cell membrane protein expressed in the luminal membrane of airway epithelial cells. However, panterin expression 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 the common cold caused by rhinovirus. Upregulation of panterin was observed when these cells were cultured in primary airway epithelial cells alongside IL-4, IL-13, and IL-17A. Interestingly, panterin knockout (KO) ameliorated airway inflammation in all mouse models of COPD, allergic rhinitis, asthma, Bordetella pertussis infection, and rhinovirus infection. The pathophysiological role of panterin in airway infection remains unclear. However, new evidence suggests that panterin is involved in the preservation of airway surface fluid (ASL) volume and the regulation of mucus production in inflammatory airway diseases.

[0003] Compared with the WT mouse control group, IL-13 significantly increased ASL volume in primary mouse tracheal epithelial cell cultures in panterin-knockout (KO) mice. The IL-13-induced increase in ASL volume in primary human nasal epithelial (HNE) cell cultures from deaf patients carrying the panterin mutant (DFNB4) was significantly higher than in normal controls. Furthermore, in primary cultures of human bronchial epithelial cells, inhibition of panterin by panterin inhibitors significantly increased IL-13-induced ASL volume. These findings suggest that downregulation of panterin may play a beneficial role in regulating ASL volume homeostasis in inflammatory airway diseases.

[0004] Excessive mucus production is a common feature of inflammatory airway diseases such as asthma and chronic obstructive pulmonary disease (COPD). Panterin overexpression significantly increased MUC5AC gene expression in human lung cancer NCI-H292 cell line and mouse lung tissue. While IL-3 treatment significantly increased MUC5AC gene expression in HNE cells of normal subjects, IL-13-induced MUC5AC upregulation completely disappeared in HNE cells from deaf patients carrying the panterin mutant. These results suggest that downregulating panterin may be beneficial in the treatment of asthma and COPD. Summary of the Invention

[0005] Technical issues

[0006] This invention is based on the discovery that certain compounds can serve as panterine inhibitors with the potential to treat respiratory diseases. It is based on the discovery that some compounds or newly designed compounds identified through cell-based HTS screening for the identification of small-molecule panterine inhibitors can provide potential therapeutic treatments for respiratory diseases (inflammatory airway diseases) such as asthma, acute or chronic bronchitis, allergic rhinitis, acute respiratory infections, cystic fibrosis, acute respiratory distress syndrome (ARDS), acute lung injury (ALI), or chronic obstructive pulmonary disease (COPD). This invention is based on the discovery that some small molecules can demonstrate that panterine downregulation reduces IL-13-induced upregulation of MUC5AC gene expression in HNE cells differentiated from normal subjects. Furthermore, in an ovalbumin (OVA)-induced allergic asthma mouse model, some molecules have been used as panterine inhibitors to improve airway inflammation.

[0007] Technical solution

[0008] On one hand, the present invention provides compounds represented by the following chemical formula 1, their E- or Z-isomers, their optical isomers, mixtures of the two isomers, their precursors, their pharmaceutically acceptable salts, or their solvates:

[0009] Chemical Formula 1

[0010]

[0011] In chemical formula 1,

[0012] V 1 and V 2 It is aryl, heteroaryl, C3-C7 cycloalkyl, heterocycloalkyl, C1-C6 alkyl, C1-C6 heteroalkyl, C2-C 10 alkenyl, C0-C3 methylene hydrazine, C2-C 10 Alkyne group, S(O) i (C1-C6 alkyl), OS(O) i(Aryl), S(O) i NR 3 R 4 C(O)R 3 OR 3 OCOR 3 NR 3 C(O)OR 4 NR 3 C(O)R 4 C(O)NR 3 R 4 Or NR 3 R 4 Furthermore, aryl, heteroaryl, C3-C7 cycloalkyl, heterocycloalkyl, C1-C6 alkyl, C1-C6 heteroalkyl, C2-C 10 alkenyl, C0-C3 methylene hydrazine, C2-C 10 Alkyne group, S(O) i (C1-C6 alkyl), OS(O) i (Aryl), S(O) i NR 3 R 4 C(O)R 3 OR 3 OCOR 3 NR 3 C(O)OR 4 NR 3 C(O)R 4 C(O)NR 3 R 4 and NR 3 R 4 Optionally, one or more of the groups are independently selected from hydrogen, oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, sulfonyl, aryl, C1-C 10 Alkyl, C3-C7 cycloalkyl, heteroaryl, heterocycloalkyl, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i NR 3 (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 OCR 3 F2, OCOR 3 NR 3 C(O)OR 4 NR 3 C(O)R 4C(O)NR 3 R 4 and NR 3 R 4 The groups are substituted, and the aryl, C1-C1 groups are substituted. 10 Alkyl, C3-C7 cycloalkyl, heteroaryl, heterocycloalkyl, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i NR 3 (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 OCR 3 F2, OCOR 3 NR 3 C(O)OR 4 NR 3 C(O)R 4 C(O)NR 3 R 4 and NR 3 R 4 Optionally, one or more of the radicals are independently selected from hydrogen, oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, aryl, C1-C2. 10 Alkyl, aryl, C3-C7 cycloalkyl, heteroaryl, heterocycloalkyl, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i (Aryl), S(O) i (Heteroaryl), S(O) i NR 3 (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 OCR 3 F2, OCOR 3 NR 3 C(O)OR 4 NR 3 C(O)R 4 C(O)NR 3 R 4 and NR 3 R 4 Group substitution, and

[0013] i and j are independently 0, 1, or 2, and

[0014] R 1 and R 2 Independently selected from hydrogen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, sulfonyl, aryl, C1-C 10 Alkyl, heteroaryl, heterocyclic, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 The group consists of aryl, heteroaryl, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 Optionally, one or more of the groups are independently selected from oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, aryl, heteroaryl, heterocyclic, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 Group substitution, and

[0015] R3 and R 4 Independently selected from hydrogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, sulfonyl, aryl, aryl (C1-C2) 10 Alkyl groups, C1-C6 10 Alkyl, heteroaryl, heteroaryl (C1-C2) 10 Alkyl groups, C1-C6 10 Alkyl heteroaryl, C1-C 10 The group consisting of alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, heterocyclic and trifluoromethyl groups, and aryl, aryl (C1-C6) 10 Alkyl groups, C1-C6 10 Alkyl, heteroaryl, heteroaryl (C1-C2) 10 Alkyl groups, C1-C6 10 Alkyl heteroaryl, C1-C 10 Alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, and heterocyclic groups are optionally selected independently by one or more groups selected from hydrogen, oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, aryl, heteroaryl, heterocyclic, C1-C6 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 Substitution of groups, or

[0016] R 3 and R 4 It can be cyclized into a 4- to 10-membered carbocyclic, heterocyclic, aromatic, or heteroaromatic ring, and one of the carbocyclic, heterocyclic, aromatic, or heteroaromatic rings may optionally be selected independently from one or more elements, including hydrogen, oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, aryl, C1-C2. 10 Alkyl, heteroaryl, heterocyclic, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 Group substitution, and

[0017] A 1 Represented as or and

[0018] X 1 and X 2 Independently select O, S, CHR 4 and NR 4 The group formed, and

[0019] X 3 Choose free hydrogen, OR 3 aryl, heteroaryl, C3-C7 cycloalkyl, heterocycloalkyl, C1-C6 alkyl, C1-C6 heteroalkyl, C2-C 10 alkenyl, C0-C3 methylene hydrazine, C2-C 10 Alkyne group, S(O) i (C1-C6 alkyl), S(O) i NR 3 C(O)R 3 OC(O)R 3 (O)COR 3 NR 3 C(O)OR 3 NR 3 C(O)R 3 C(O)NR 3 and NR 3 R 4 The group formed, and OR 3 aryl, heteroaryl, C3-C7 cycloalkyl, heterocycloalkyl, C1-C6 alkyl, C1-C6 heteroalkyl, C2-C 10 alkenyl, C0-C3 methylene hydrazine, C2-C 10 Alkyne group, S(O) i (C1-C6 alkyl), S(O) i NR 3 C(O)R 3 OC(O)R 3 (O)COR 3 NR 3 C(O)OR 3 NR 3 C(O)R 3 C(O)NR 3 and NR3 R 4 One of them is optionally selected independently from hydrogen, oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, sulfonyl, aryl, C1-C 10 Alkyl, C3-C7 cycloalkyl, heteroaryl, heterocycloalkyl, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i NR 3 (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 OCR 3 F2, OCOR 3 NR 3 C(O)OR 4 NR 3 C(O)R 4 C(O)NR 3 R 4 and NR 3 R 4 The groups in, and aryl, C1-C 10 Alkyl, C3-C7 cycloalkyl, heteroaryl, heterocycloalkyl, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i NR 3 (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 OCR 3 F2, OCOR 3 NR 3 C(O)OR 4 NR 3 C(O)R 4 C(O)NR 3 R 4 and NR 3 R 4 Optionally, one or more of the groups are independently selected from hydrogen, oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, sulfonyl, aryl, C1-C 10 Alkyl, aryl, C3-C7 cycloalkyl, heteroaryl, heterocycloalkyl, C1-C 10Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i (Aryl), S(O) i (Heteroaryl), S(O) i NR 3 (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 OCR 3 F2, OCOR 3 NR 3 C(O)OR 4 NR 3 C(O)R 4 C(O)NR 3 R 4 and NR 3 R 4 Group substitution, and

[0020] R 5 and R 6 Independently selected from hydrogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, sulfonyl, aryl, C1-C 10 Alkyl, heteroaryl, heterocyclic, C1-C 10 The group consisting of alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, heterocyclic, aryl, heteroaryl, and trifluoromethyl, wherein alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, C1-C6... 10 One of the alkylaryl and heteroaryl groups is optionally selected from one or more groups, and is independently selected from hydrogen, oxygen, halogen, cyano, azathiol, nitro, trifluoromethyl, trifluoromethoxy, aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclic, C1-C2. 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i (Aryl), S(O) i (Heteroaryl), S(O) i NR 3 R 4 C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R4 and NR 3 R 4 One or more groups in the group, and aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclic, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i (Aryl), S(O) i (Heteroaryl), S(O) i NR 3 R 4 C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 One of the groups is optionally selected from one or more independently selected from hydrogen, oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclic, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i (Aryl), S(O) i (Heteroaryl), S(O) i NR 3 R 4 C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 One or more groups in, or

[0021] R 5 and A 1 It can be cyclized into a 4- to 10-membered carbocyclic, heterocyclic, aromatic, or heteroaromatic ring, and one of the carbocyclic, heterocyclic, aromatic, or heteroaromatic rings may optionally be selected independently from one or more elements, including oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, aryl, C1-C2, etc. 10Alkyl, heteroaryl, heterocyclic, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 Substitution of groups in [the group].

[0022] On the other hand, the present invention provides a pharmaceutical composition for the prevention or treatment of respiratory diseases (inflammatory airway diseases), comprising at least one compound, 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 as an active ingredient. In some embodiments, the composition can inhibit, prevent, improve, or treat respiratory diseases (inflammatory airway diseases).

[0023] On the other hand, 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 and a pharmaceutically acceptable carrier.

[0024] On the other hand, the present invention provides the use of a compound represented by chemical formula 1 and a pharmaceutical composition thereof as a panterine inhibitor.

[0025] On the other hand, the present invention provides a compound represented by chemical formula 1 and a pharmaceutical composition thereof that specifically controls chloride ion channels.

[0026] On the other hand, the present invention provides a compound represented by chemical formula 1 and a pharmaceutical composition thereof, which retains the volume of airway surface fluid (ASL) and reduces mucin separation.

[0027] In another aspect, the present invention provides use for one or more respiratory diseases (inflammatory airway diseases), said respiratory diseases being 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).

[0028] Other aspects and advantages of the invention will be apparent to those skilled in the art from the detailed embodiments and accompanying drawings.

[0029] Beneficial effects

[0030] According to the present invention, the novel compound can be used as a panterine inhibitor, and therefore, it can be used as a composition for the prevention, treatment or improvement of respiratory diseases (inflammatory airway diseases, especially asthma or acute lung injury). Attached Figure Description

[0031] Figure 1 The principle and results of cell-based high-throughput screening are shown.

[0032] Figure 2 The YFP fluorescence tracing diagram shows the effect of F56 on the Cl- of panterin in CHO-K1-YFP cells that stably express human panterin. - / I - Examples of the inhibitory effect of exchange activity.

[0033] Figure 3 This demonstrates the effect of F56 on Pantelin-mediated Cl - / HCO3 - Inhibition of exchange activity. This is achieved through pantelin-mediated Cl... - / HCO3 - Exchange, using high concentrations of Cl - It induces a decrease in intracellular pH and leads to a decrease in YFP fluorescence. Pretreatment with a specified concentration of pantelin inhibitor for 10 minutes.

[0034] Figure 4(a) shows the inhibition of pantelin-mediated Cl - / I - and Cl - / HCO3 - Summary of F56 dose response with exchange activity (mean ± SE, n = 4–5).

[0035] Figure 4(b) shows the inhibition of pantelin-mediated Cl - / I - Cl - / SCN - Cl - / HCO3 - and Cl - / OH - Summary of F10 dose response for exchange activity (mean ± SE, n = 3).

[0036] Figure 5(a) shows how the novel panterin inhibitor F56 blocks the panterin-mediated cellular response in IL-4-stimulated HNE cells. (A) Chemical structure of F56. (B) Effect of F56 on panterin-mediated Cl- - / SCN - Inhibition of exchange activity (mean ± SE, n = 5). (C)F56 inhibits pantelin-mediated Cl- - / I - Cl - / SCN - Cl - / HCO3 - and Cl - / OH - Dose-response of exchange activity (mean ± SE, n = 4–5). (D) Representative protein expression of pantelin (PDS) in untreated and IL-4-treated HNE cells. (E) Pantelin-mediated Cl- exchange activity in IL-4-treated HNE cells. - / HCO3 - Exchange. (F)ΔpH i Initial rate of change at / min (mean ± SE, n = 3-4). (G) Cl in IL-4 treated HNE cells exposed to F56 after Cl - / HCO3 - Exchange activity. (H) Representative protein expression of pantelin and AN01 in IL-4 treated HNE cells (mean ± SE, n = 5). (I) Determination of PDS mRNA expression levels in IL-4 treated HNE cells by real-time PCR (mean ± SE, n = 5). *P < 0.05, **P < 0.01, ***P < 0.001. NTC, untransfected cells.

[0037] Figure 5(b) shows the effect of F10 on the mRNA expression level of PDS. The mRNA expression level of PDS within 120 hours after F10 treatment was determined by real-time quantitative PCR in IL-4 treated HNE cells.

[0038] Figure 6(a) shows the panterin-mediated Cl-mediated flow in CHO-K1 cells expressing human panterin. - / Base exchange activity. (A) Pantelin-mediated Cl - / I - Exchange activity (mean ± SE, n = 5). Pretreatment with a specified concentration of F56 for 10 min. (B, C) Representative tracking of intracellular pH. Application of a chlorine-free solution via pantelin-mediated Cl- exchange. - / HCO3 - and Cl - / OH -Exchange activity induces intracellular alkalization. Pretreatment with a specified concentration of F56 for 10 minutes was performed. (DF) Pantelin-mediated Cl- levels were measured in CHO-K1 cells expressing human pantelin. - / I - Exchange activity. A specified concentration of F56 and PDS is applied. inh -A01 and PDS inh -C01 Pretreatment for 10 minutes. (G)F56, PDS inh -A01 and PDS inh -C01 dose response summary (mean ± SE, n = 3). (H, I)F56, PDS inh -A01 and PDS inh Effects of -C01 on the cell viability of NIH3T3(H) and CHO-K1(I) cells. Using F56 and PDS... inh -A01 and PDS inh Cells were treated with -C01 for 24 hours. Cell viability was measured by MTS colorimetric analysis (mean ± SE, n = 3). NTC, untransfected cells.

[0039] Figure 6(b) shows the effect of F10 on cell viability of NIH3T3 and CHO-K1 cells. Cells were treated with F10 for 24 hours. Cell viability was determined by MTS colorimetric analysis (mean ± SE, n = 3).

[0040] Figure 7(a) shows the properties of F56. (A) The effect of F56 on mouse pantelin (mPDS)-mediated Cl- in CHO-K1 cells was measured. - / I - Inhibition of exchange activity (mean ± SE, n = 5). Pretreatment with specified concentrations of F56 for 10 min. (B) Inhibition of human pantelin (hPDS)-mediated (solid circles, Fig. (a), C) and mPDS-mediated (hollow circles) Cl - / I - A summary of the dose-response relationship of exchange activity. (CF) shows a representative tracking of intracellular pH in CHO-K1 cells expressing SLC26A3, SLC26A6, SLC26A7, and SLC26A9. Chlorine-free solution was applied via Cl... - / HCO3 - Exchange induces intracellular alkalization. Pretreatment with a specified concentration of F56 for 10 min. (G) The effect of F56 (100 μM) on CFTR chloride channel activity was measured in FRT cells expressing human CFTR. CFTR currents were activated by 20 μM trichodin and by 10 μM CFTR. inh-172 inhibition. (H) Effect of F56 (100 μM) on AN01 chloride channel activity in FRT cells expressing human AN01. F56 was added 10 min before AN01 was activated by 100 μM ATP. (Right) Summary of peak currents (mean ± SE, n = 3). (I) Effect of F56 on hERG (Kv11.1) potassium channel activity in HEK293T cells expressing human Kv11.1 (mean ± SE, n = 4). Pretreatment with the specified concentration of F56 for 10 min. hERG channels were inhibited by 50 μM cisapride. (J) Effect of F56 on hERG (Kv11.1) potassium channel activity in HEK293T cells expressing human 5-HT 2A Measuring F56-5-HT in FRT cells 2A Effects on channel activity (mean ± SE, n = 4). Pretreatment with a specified concentration of F56 for 10 minutes. 5-HT 2A The channel was activated by 10 μM 5-HT and inhibited by 10 μM ketoserine. ***P<0.001. NTC, untransfected cells.

[0041] Figure 7(b) illustrates the effect of F10 on the activity of CFTR, ANO1, and hERG channels. (A) The effect of F10 (30 μM) on CFTR channels was measured in FRT cells expressing human CFTR and the mutant YFP. CFTR is activated by 20 μM trichodin and by 10 μM CFTR. inh -172 inhibition. (B) Effect of F10 (30 μM) on AN01 channel activity was measured in FRT cells expressing human and mutant YFP. AN01 is activated by 100 μM ATP and activated by 10 μM T16A. inh -A01 inhibition. (C) Effect of F10 (30 μM) on hERG (Kv11.1) potassium channel activity in HEK293T cells expressing human Kv11.1 was measured (mean ± SE, n = 4). hERG channels were inhibited by 50 μM cisapride.

[0042] Figure 8(a) shows the effect of F56 on pantelin and other ion channels in HNE cells. (A) Representative tracking of intracellular pH in HNE cells treated (grey line) and treated with IL-4 (10 ng / ml). Pantelin was inhibited by F56 (50 mM). (B) Real-time quantitative PCR determination of pantelin mRNA expression levels (mean ± SE, n = 3) in IL-4 treated HBE cells at specified time points following F56 (30 mM) treatment. (C) Measurement of Cl in IL-4 treated HBE cells. - / HCO3 -Exchange activity. F56 pretreatment for 5 min. Short-circuit current recording of (D,E)HNE cells. Representative tracer analysis shows the effect of F56 on ENaC, CFTR, and CaCC channel activity. Channel activity was measured with or without IL-4 after 48 hours of exposure to F56 (30 mM). Amiloride (100 μM), salvia miltiorrhiza (20 μM), and CFTR were also measured. inh -172 (10 μM) and ATP (100 μM) were added to the top bath. (F) At the specified time after F56 (30 mM) treatment, the expression levels of ANO1, CFTR and ENaC mRNA in IL-4-treated HNE cells were determined by real-time quantitative PCR (mean ± SE, n = 5) *P < 0.05, **P < 0.01, ***P < 0.001.

[0043] Figure 8(b) shows the effect of F10 on the mRNA expression levels of ANO1, CFTR, and ENaC. The mRNA expression levels of ANO1, CFTR, and ENaC were determined by real-time quantitative PCR within 120 hours after F10 treatment in IL-4-treated HNE cells.

[0044] Figure 9(a) shows that F56 inhibits pantelin / SCN in an OVA-induced allergic asthma mouse model. - / NF-κB pathway improves airway inflammation. (A) Airway resistance in animals sensitized and challenged by OVA. (B) IL-4 and F56 in HNE cells for SCN. - Effects of transport (mean ± SE, n = 5). (C) Levels of Duox1 and Duox2 in IL-4-treated HNE cells after F56 treatment by real-time PCR (mean ± SE, n = 3). (D) Results of protein expression levels in IL-4-treated cells with or without F56. (Right) Comparison of relative protein mass of phosphate-p65 (mean ± SE, n = 3). (E) Airway resistance to methacholine stimulation in OVA-sensitized mice (mean ± SE, n = 3–4). (F) Representative PAS staining of airway tissue. Size bar, 100 μm. *P < 0.05, **P < 0.01.

[0045] Figure 9(b) shows the effect of F10 on airway resistance in animals sensitized and challenged with OVA.

[0046] Figure 10This study illustrates the ameliorative effect of F56 on allergic airway inflammation in an asthmatic mouse model. (A) Protocol and time course for inducing allergic airway inflammation in wild-type mice after allergen challenge. (B) Inflammatory cells from bronchoalveolar lavage fluid (BALF) separated by centrifugation and stained with Diff-Quik staining reagent. Cell counts were quantified by light microscopy (mean ± SE, n = 10). (C) Representative histology of airways stained with hematoxylin and eosin. Size bar, 100 μm. (D) Periodic acid-Schiff (PAS) staining of airway portions from OVA-sensitized / challenged mice treated with vector, OVA, and F56 (10 mg / kg). Size bar, 50 μm. (E) Summary of inflammation scores (mean ± SE, n = 4). (F) Serum OVA-specific IgE levels (mean ± SE, n = 4). (G) Comparison of NF-κB expression in the lungs of NF-κB reporter mice using the IVIS system. Lungs were isolated from mice and fluorescence imaging was performed using IVIS spectroscopy (Ex570, Em620). The mean fluorescence density of the images was analyzed using Image 4.3.1 software. *P<0.05.

[0047] Figure 11 This study demonstrates the improvement of allergic asthma by F56 in an established allergic asthma model. (A) Protocol and time course for inducing allergic airway inflammation in wild-type mice following allergen antigen challenge. F56 (10 mg / kg, 3 times / day) was administered after three OVA challenges. (B) Airway responsiveness in an OVA-induced asthma model as measured by whole-body plethysmography. Penh was measured based on increasing doses of methacholine (mean ± SE, n = 4). (C) Representative PAS staining of airway tissue. Size strips, 100 μm. (D) Summary of PAS-positive cells (mean ± SE, n = 4). *P < 0.05.

[0048] Figure 12(a) illustrates the effect of F56 on mucosal production and ASL volume control in IL-4-treated HNE cells. (A) MUC5AC mRNA expression levels were measured at specified time points following F56 (30 mM) treatment in IL-4 (10 ng / ml)-treated HNE cells (mean ± SE, n = 4). (B) Periodic acid-Schiff (PAS) staining showed goblet cell proliferation in IL-4 and IL-13 (10 ng / ml)-treated HNE cells with or without F56 (30 M) on day 7. (C) Total volume of ASL and fluid meniscus was measured in HNE cells expressing wild-type (wt) or mutant (mt) PDS. HNE cells were treated with IL-4 (10 ng / ml, 48 h) in or without 30 mM F56 (mean ± SE, n = 3–4). (D) Representative images of transwell inserts from normal HNE cells. Arrows indicate the fluid meniscus of HNE cells treated with IL-4 (10 ng / ml, 48 h) in the presence or absence of 30 mM F56. (E) Representative protein expression results of pantelin in HNE cells treated with IL-4 and IL-13. (F) Measurement of total volume of ASL and fluid meniscus. HNE cells treated with IL-13 (10 ng / ml, 48 h) in the presence or absence of 30 mM F56 (mean ± SE, n = 3–4). *P < 0.05, ***P < 0.001. Size bar, 30 μm.

[0049] Figure 12(b) shows the effect of F10 on MUC5AC mRNA expression levels. MUC5AC mRNA expression levels in IL-4-treated HNE cells were measured by real-time quantitative PCR within 120 hours after F10 treatment.

[0050] Figure 13 The effects of F56 on hearing thresholds and plasma thyroid hormone levels are shown. (A) Representative examples of ABR waveforms in control and F56 (10 mg / kg / yl, 7 days) treated mice. (B) Summary of hearing thresholds (mean ± SE, n = 8–10). (C) Serum levels of T3 and T4 measured in F56 (10 mg / kg / yl, 7 days) treated and untreated mice (mean ± SE, n = 4). (D) Representative power tracking showing the airway smooth muscle (ASM) contractile response in the rat tracheal rings. F56 (30 mM) was applied after ASM contraction was induced by submaximal concentration (300 nM) of carbachol (CCh). ASM relaxation was induced by trichodin and IBMX.

[0051] Figure 14This is a schematic diagram illustrating the potential role of pantelin inhibitors in airway inflammation. Pantelin inhibitors can reduce NF-κB activation by blocking SCN transport in the airway epithelium and enhance mucosal clearance by inhibiting pantelin-mediated ASL deficiency upregulated in airway inflammation.

[0052] Figure 15 The study demonstrated that pantelin deficiency attenuated LPS-induced lung injury in mice. LPS (10 mg / kg) or a carrier (PBS) was administered intranasally to wild-type (WT) and pantelin-free (PdS) ​​mice. - / - (A) Analysis of total number of bronchoalveolar lavage (BAL) cells and BAL protein concentration in WT mice within 48 hours after administration of LPS or PBS. (B) Representative images of H&E staining of lung tissue within 48 hours after LPS or PBS administration (×400), bar size: 50 μm. (C) Analysis of total number of BAL cells and BAL protein concentration within 48 hours after LPS or PBS administration. (D) Representative images of H&E staining of lung tissue within 48 hours after LPS or PBS administration (×400), bar size: 50 μm. (E) Changes in mouse body weight. (F) Representative protein expression results of pantelin in lung lysates from LPS-untreated and LPS-treated WT mice. Data provided were analyzed by Student's unpaired two-tailed t-test, mean ± SEM (n = 6–8 mice per group), *P < 0.05, **P < 0.01, ***P < 0.001.

[0053] Figure 16 This study demonstrates the effect of a novel panterin inhibitor (F56) on blocking panterin activity in human alveolar epithelial cells. (A) Chemical structure of the panterin inhibitor F56. (B) Representative protein expression results of panterin (PDS) in human alveolar epithelial cells (hAEC). (C) Measurement of PDS mRNA levels in hAEC by real-time quantitative PCR (mean ± SE, n = 3). (D) Effect of F56 on human wild-type panterin-mediated Cl- in hAEC expressing human panterin. - / SCN - Inhibition of exchange activity (mean ± SE, n = 10). Pretreatment with the specified concentration of F56 for 10 min. (Right) Dose response summary. (E) DUO×2 mRNA levels as measured by real-time quantitative PCR (mean ± SE, n = 3–4). *P < 0.05, **P < 0.01, relative to control group. Unpaired two-tailed t-test by students.

[0054] Figure 17The following diagram illustrates the inhibition of LPS-induced acute lung injury phenotype by F56 in mice. (A) Intraperitoneal injection of F56 (10 mg / kg) 1 hour prior to LPS treatment. (B) Total BALF cell count. (C) BALF protein concentration. (D) Representative image of H&E stained lung tissue (×400), size bar: 50 μm. (E) Lung injury score. (F) Intraperitoneal injection of F56 (10 mg / kg) at 6 and 12 hours after LPS inhalation. (G) Total BALF cell count. (H) BALF protein concentration. (I) Lung injury score. Data presented were analyzed by one-way ANOVA and Bonferroni post-hoc test, mean ± SEM (n = 10-12 mice per group), ***P < 0.001.

[0055] Figure 18 This shows SCN in mice with or without F56. - Triggered LPS-induced lung injury. (A) Total BALF cell count. WT mice were treated with LPS (10 mg / kg, intranasal), F56 (10 mg / kg, ip), NaOH (100 mM, intranasal), NaHCO3 (100 mM, intranasal), and NaSCN (100 mM, intranasal). (B) BALF protein concentration. (C) Lung injury score. (Right) Representative lung tissue stained with H&E (×400), size bar: 50 μm. (D) BALF protein concentration in pantelin-deficient mice. Pantelin-deficient mice were treated with LPS-treated with NaSCN (100 mM, intranasal). (Right) Centrifuged smear of representative mouse BALF cells stained with Diff-Quik staining. Compared with LPS or NaSCN alone, LPS+NaSCN exposure increased the number of inflammatory cells, particularly neutrophils (red arrows). Black arrows indicate macrophages (x200), size bar: 100 μm. The data were analyzed by one-way ANOVA and Bonferroni post-hoc test. Mean ± SEM (5-6 mice per group), *P<0.05, **P<0.01, ***P<0.001.

[0056] Figure 19This study demonstrates how F56 blocks the NF-κ pathway and reduces the levels of pro-inflammatory cytokines in LPS-induced acute lung injury. (A) Representative images of lungs from NF-κ / SPC-Cre mice exposed to LPS (10 mg / kg) and treated with F56 (10 mg / kg) or a carrier. IVIS image fluorescence is expressed as radioefficiency. (B) Mean fluorescence was quantified by analyzing the region of interest using Living Image software. Data provided are mean ± SEM (n = 9–10 mice per group). (C) Representative protein expression results in lung lysates. (D) Relative protein levels measured by density measurements of pantelin and phosphate-Iκ (mean ± SEM, n = 6 per group). (EH) Levels of IL-1βCXCL2 / MIP-2, IL-6, and TNF-α were measured in lung tissue lysates by ELISA. The data were analyzed by one-way ANOVA and Bonferroni post-hoc test, mean ± SEM (n = 7-8 mice per group), *P<0.05, **P<0.01, **P<0.001.

[0057] Figure 20 The panterine levels in human BALF are shown. Patients with ARDS due to pneumonia showed increased panterine levels compared with control patients (uninfected). Panterine levels were measured from human BALF supernatant by ELISA (control group n=25, ARDS n=41). Analysis by Student's unpaired two-tailed t-test showed *P<0.05, **P<0.01, ***P<0.001.

[0058] Figure 21 A schematic diagram illustrating the role of pantelin and its inhibitors in LPS-induced lung injury is shown. (SCN) - Active transport of panthenol from the apical surface of the alveolar epithelium to the lung cavity. SCN - Catalyzed by peroxidase and H2O2 to OSCN - The resulting OSCN - It activates NF-κ and induces the release of inflammatory cytokines, neutrophil infiltration, and subsequent lung injury. The panterine inhibitor F56 blocks SCN. - Transepithelial transport, thereby inhibiting OSCN - Induced NF-κ activation and subsequent ALI onset. Detailed Implementation

[0059] 1. Definition

[0060] Unless otherwise defined, all technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this relates. The following references provide general definitions for many of the terms used herein: The Cambridge Dictionary of Science and Technology (Walker, 1988); The Glossary of Genetics, 5th edition, R. Rieger et al., Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). As used herein, unless otherwise stated, the following terms have the meanings referred to below.

[0061] Unless otherwise specified or clear from the context, the term “or” as used herein is to be understood as included.

[0062] Unless specifically stated or clear from the context, the term "about" as used herein is to be understood as being within the generally accepted range in the field, such as within two standard deviations of the mean. "About" can be understood as being within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the specified value. Unless the context clearly indicates otherwise, all numerical values ​​provided herein are modified by the term "about".

[0063] The terms “activator,” “drug,” and “drug formulation” are used interchangeably herein to refer to a chemical material or compound that, when administered to a subject (e.g., any animal, including humans or non-human animals) in any of the ways described herein, induces a desired pharmacological effect (e.g., reduction of inflammation).

[0064] As used herein, “additive” can refer to any additional ingredient that can be added to the compositions and chemical formulas described herein. For example, assuming that the additional ingredient is pharmaceutically acceptable for the specific condition being treated, the additive may 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), isotonic adjusters (e.g., one or more isotonic adjusters), thickeners (e.g., one or more thickeners), suspending agents (e.g., one or more suspending agents), binders (e.g., one or more binders), viscous agents (e.g., one or more viscous agents), etc. Furthermore, additives may include therapeutic agents and drug delivery modifiers, as well as enhancers (e.g., calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starch, gelatin, cellulose, methylcellulose, sodium carboxymethylcellulose, glucose, hydroxypropyl-β-cyclodextrin, polyvinylpyrrolidone, low-melting-point waxes, and ion exchange resins), and combinations of any two or more of these. 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 ed. (2003) and 21st ed. (2005), respectively, which are incorporated herein by reference. The additives described herein may be used in any suitable pharmaceutical preparation.

[0065] As used herein, the term "administration" means oral administration, suppository administration, local contact intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, intranasal, intravitreal, or subcutaneous administration, or continuous-release devices, such as small osmotic pumps implanted in the subject. Administration can be administered via any route, including parenteral and transmucosal routes (e.g., oral, intranasal, intrapulmonary, rectal, buccal, vaginal, intraocular, and dermal).

[0066] The terms "analyte" and "derivative" are used interchangeably herein, referring to a compound that has the same core as a parent compound but differs in the bonding sequence, in the absence or presence of one or more atoms and / or groups of atoms and combinations thereof. The derivative may differ from the parent compound in one or more substituents present on the core, which may contain, for example, one or more atoms, functional groups, or substructures. Furthermore, the derivative may differ from the parent compound in the bonding sequence between the core atoms. Typically, the derivative can be predicted, at least theoretically, to be formed from the parent compound through chemical and / or physical processes.

[0067] As used herein, "antioxidant" can refer to any artificial or natural substance capable of preventing or delaying a certain type of damage and / or oxidation. Antioxidants are found in many foods, including fruits and vegetables. They are also available as dietary supplements. Exemplary antioxidants may include beta-carotene, lutein, lycopene, selenium, vitamin A, vitamin C, and vitamin E. Other antioxidants known to those skilled in the art may also be used. The antioxidants described herein may be used in any suitable amount.

[0068] "Co-administration" means administering a compound or composition described herein immediately before or immediately after administration of an additional therapeutic agent, activator, or additive described herein. The compounds or compositions of the present invention may be administered to a patient alone or co-administered. Co-administration is interpreted as including administering the compound, alone or in combination (one or more compounds or agents), simultaneously or sequentially. If desired, the agent may also be combined with other active substances.

[0069] In this document, terms such as “comprising,” “including,” “containing,” and “having” may have meanings belonging to them and may mean “including,” “containing,” etc.; and “consistently composed of” or “consistently made of” may have similar meanings related to them, and these terms are open-ended, allowing for the existence of more than those listed, unless the basic or novel features listed are altered by the existence of more of those listed, but excluding instances of the prior art.

[0070] As used herein, "concurrent administration" includes at least part of an overlap in duration. For example, when two agents (e.g., any agent or class of agents that are biologically active as described herein) are administered concurrently, their administration occurs within a desired time period. Administration of the formulations may begin and end on the same day. Furthermore, administration of one formulation may precede administration of the second formulation, provided that both formulations are taken at least once on the same day. Similarly, administration of one formulation may be extended after administration of the second formulation, provided that both formulations are taken at least once on the same day. To include concurrent administration of biologically active agents / formulations, it is not necessary to take them at the same time every day.

[0071] As used herein, "effective amount" or "therapeutic effective amount" refers to an amount sufficient to influence the desired biological effect, such as beneficial outcomes (including clinical outcomes). Therefore, "effective amount" is dependent on its application context. Effective amounts can vary based on factors known in the art, such as the disease state, age, sex, and weight of the individual being treated. Dosage can be divided into several daily doses, or the dosage can be reduced proportionally according to the urgency of the treatment situation. Furthermore, the compositions / formulations of the present invention can be administered frequently as needed to achieve therapeutic doses.

[0072] The term "gel" as used in this article can refer to a non-flowing liquid but not a solid, i.e., a semi-solid material. Gels can be formed from natural or synthetic substances. Gels are not aligned, but rather slightly aligned, exhibiting birefringent liquid crystal properties. Gels can be used for topical drug delivery.

[0073] The term “respiratory disease” as used in this article has common medical meanings, including, but not limited to, asthma, acute or chronic bronchitis, allergic rhinitis, acute upper respiratory tract infection, cystic fibrosis, acute respiratory distress syndrome (ARDS), acute lung injury (ALI), or chronic obstructive pulmonary disease (COPD) and diseases and disorders closely related to the respiratory system.

[0074] As used herein, the term "inhibition" means prevention, reduction, mitigation, or cessation. In one embodiment, a composition or compound is considered to inhibit the activity of at least one protein (e.g., panterine) when the amount or rate of a process or response occurring in the presence of the compound or composition is reduced by at least about 10% compared to the amount or rate in the absence of the compound or composition. In another embodiment, a composition or compound is considered to inhibit the process or response when the amount or rate of a process or response occurring in the presence of the compound or composition is reduced by at least about 20% compared to the amount or rate in the absence of the compound or composition. In other embodiments, a composition or compound is considered to inhibit one or more proteins (e.g., panterine) when the amount or rate of a process or response 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% compared to the amount or rate in the absence of the compound or composition. In other embodiments, a compound or composition is considered to inhibit the activity of one or more proteins, i.e., to prevent their development.

[0075] The term "intermittent dosing" as used in this article includes the period during which the formulation is administered (which can be considered the "first dosing period"), the subsequent period during which the formulation is not ingested or is ingested at a lower dose (which can be considered a "discontinuation period"), and the period during which the formulation is subsequently administered again (which can be considered the "second dosing period"). Generally, during the second dosing period, the dosage level of the formulation is consistent with the dosage level administered during the first dosing period, but may be increased or decreased as needed medically.

[0076] The "jelly" according to the present invention is composed of a gel, which is a semi-solid system consisting of a suspension composed of inorganic small particles or organic macromolecules and partially permeated by a liquid having a highly viscous matrix, typically an aqueous liquid.

[0077] The term "liquid" as used herein refers to a form of administration consisting of a liquid composition. Liquids may splash; they flow and behave in a container at room temperature. Liquids exhibit Newtonian or pseudoplastic flow behavior.

[0078] In one embodiment, the term "semi-liquid" as used herein may have the properties of liquids and other formulations (i.e., suspensions, emulsions, solutions, creams, gels, jelly, etc.).

[0079] As used in this article, the term "ointment" can refer to a highly viscous liquid or semi-liquid preparation that can be used for the therapeutic treatment of a disease, syndrome or symptom.

[0080] As used herein, “pharmaceutically acceptable carriers” include any and all physiologically suitable solvents, dispersion media, coatings, antimicrobial and antifungal agents, isotonic agents, and absorption delay agents. The type of carrier may be selected based on the intended route of administration. Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the immediate preparation of sterile topical solutions or dispersions. The use of such media and reagents for pharmaceutically active substances is well known in the art. The intended use of the compositions herein is also considered, provided that any conventional media or reagent is incompatible with the composition (e.g., the composition of Formula 1, its derivatives or analogs, or pharmaceutically acceptable salts, solvents, hydrates, or polymorphs thereof).

[0081] As used herein, “pharmaceutical carrier” or “carrier” may also include pharmaceutically acceptable carriers, excipients, or stabilizers that are non-toxic to cells or mammals at the doses and concentrations used. Physiologically acceptable carriers are typically pH-buffered aqueous solutions. Examples of physiologically acceptable carriers include buffers such as phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid; low molecular weight (less than about 10 polypeptide residues); 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; counterions that form salts, such as sodium; and / or nonionic surfactants, such as Tween. TM Polyethylene glycol (PEG) and Pluronics TM In addition, "pharmaceutical acceptable" means that it is approved for use in animals, especially humans, by federal or state regulatory agencies or by relevant agencies in countries outside the United States, or is listed in the United States Pharmacopeia or other generally approved pharmacopoeias.

[0082] The term "pharmaceutically acceptable salt or complex" refers to a salt or complex represented by a specific chemical formula 1. Examples of such salts include base addition salts formed by the reaction of compounds represented by chemical formula 1, organic or inorganic bases having metal cations selected from the group consisting of alkali metals (e.g., sodium, potassium, or lithium) and alkaline earth metals (e.g., calcium or magnesium), such as hydroxides, carbonates, or bicarbonates, or having primary, secondary, or tertiary alkylamines, but not limited thereto. Amine salts induced by methylamine, dimethylamine, trimethylamine, ethylamine, diethylamine, triethylamine, morpholine, N-methyl-D-glutamine, N,N'-bis(benzyl)-1,2-ethylenediamine, tromethamine, ethanolamine, diethanolamine, ethylenediamine, N-methylmorpholine, procaine, piperidine, piperazine, etc., are considered to be within the scope of this invention.

[0083] Furthermore, the term "salt," "salt form," or "pharmaceutically acceptable salt" as used herein may include base addition salts derived from inorganic bases (from those having a free carboxyl group or other anionic group), such as sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide, or ferric hydroxide, and base addition salts of organic bases, such as isopropylamine, trimethylamine, 2-ethylaminoethanol, histidine, procaine, etc. This salt is formed as an acid addition salt having any free cation group; for example, it typically forms with inorganic acids such as hydrochloric acid, sulfuric acid, or phosphoric acid, or with organic acids such as acetic acid, citric acid, p-toluenesulfonic acid, methanesulfonic acid, oxalic acid, tartaric acid, mandelic acid, etc. Salts herein may include amine salts formed by protonation of an amino group having an inorganic acid (e.g., hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, etc.). Additionally, salts herein include amine salts formed by protonation of an amino group having a suitable organic acid (e.g., p-toluenesulfonic acid, acetic acid, etc.).

[0084] As used herein, the terms "pH reagent" or "buffer" may refer to a compound or buffer used as a pH adjuster. This may include, but is not limited to, glycerol buffer, citrate buffer, borate buffer, acetate buffer, gluconate buffer, phosphate buffer, or citrate-phosphate buffer. pH reagents or buffers may be used in any suitable amount.

[0085] As used herein, the term "preservative" can refer to a substance or chemical substance that prevents undesirable changes to the compounds, compositions, or chemical formulas described herein. Suitable preservatives may include, for example, benzalkonium chloride, thimerosal, chlorobutanol, methylparaben, propylparaben, phenethyl alcohol, disodium sorbate edetate, polyquaternium salts, hexadecyl bromide, hexadecylpyridine chloride, benzyl bromide, EDTA, phenylmercuric nitrate, phenylmercuric acetate, thimerosal, thimerosal, acetates and mercuric phenylborate, polymyxin B sulfate, methylparaben and propylparaben, tertiary ammonium chloride, sodium benzoate, sodium propionate and sodium perborate, and other reagents or combinations thereof known to those skilled in the art. Preservatives may be used in any suitable amount.

[0086] As used herein, the terms “prevention,” “avoidance,” or “prevention” and other equivalents include those used to reduce the incidence of a syndrome, and the prevention of the development, occurrence, disruption, or avoidance of a syndrome of disease or condition. Prevention can be complete (i.e., no detectable symptoms) or partial, and therefore involves fewer observable symptoms compared to the absence of treatment. The term also includes preventative benefits. To prevent a disease or condition, the composition may be administered to a patient at risk of developing a specific disease or a patient with one or more physiological syndromes that have reported a disease, although a diagnosis of disease is not necessarily required.

[0087] The ranges provided herein should be understood as abbreviations of all values ​​within that range. For example, the range 1 to 10 is understood to include 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 combination of numbers from groups consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, or combinations of subrange numbers. Regarding subranges, particular consideration is given to “nested subranges” extending from one of the endpoints of a range. For example, overlapping subranges of the instance range 1 to 50 could 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 another direction. A range may be expressed herein as “approximately” a particular value and / or “approximately” other particular values. When expressing such a range, other aspects include a particular value and / or other particular values. Similarly, when the antecedent "about" is used to represent a value as an approximation, it should be understood that a particular value forms another aspect. It should also be understood that the endpoints of each range are significant relative to and independent of the other endpoints. Furthermore, throughout the application, it can be understood that data is provided in a variety of different formats and that these data represent endpoints and starting points and ranges of arbitrary combinations of data points. For example, when specific data point "10" and specific data point "15" are disclosed, it is considered that values ​​between 10 and 15, as well as greater than, greater than or equal to, less than, less than or equal to, and equal to, are disclosed. Furthermore, it should be understood that each unit between two specific units is disclosed. For example, when 10 and 15 are disclosed, 11, 12, 13, and 14 are also disclosed.

[0088] Other excipients considered for use in the practice of this invention are those available to those skilled in the art, such as those found in the United States Pharmacopeia Vol. XXII and the National Formulary Vol. XVII, US Pharmacopoeia Convention, Inc., Rockville, Md. (1989), the relevant contents of which are incorporated herein by reference.

[0089] According to this disclosure, a "semi-solid gel" is a semi-solid. The apparent viscosity of a semi-solid formulation may increase with increasing concentration.

[0090] As used herein, “sequential administration” includes administration of two formulations (e.g., the compounds or compositions described herein) occurring separately on the same day or not on the same day (e.g., consecutively).

[0091] According to this disclosure, a "solution" can be a clear, homogeneous liquid form of administration containing one or more chemical substances dissolved in a solvent or mixture of solvents that are miscible with each other. Because the molecules of the drug substance are uniformly dispersed in a solution, using a solution as a form of administration generally ensures uniform dosage during administration and good accuracy when the solution is diluted or otherwise mixed.

[0092] As used herein, the term "solvent" refers to an aqueous or non-aqueous liquid solvent. The choice of solvent depends particularly on the solubility of the composition and the route of administration. Aqueous solvents may consist of water alone, or may consist of water and one or more miscible solvents, and may contain dissolved solutes such as sugars, buffers, salts, or other excipients. Commonly used non-aqueous solvents include short-chain organic alcohols such as methanol, ethanol, and propanol; short-chain ketones such as acetone; and polyols such as glycerol.

[0093] "Subject" or "patient" refers to a human or non-human animal, such as a mammal. "Subject" can include any animal, including horses, dogs, cats, pigs, goats, rabbits, hamsters, monkeys, guinea pigs, rats, mice, lizards, snakes, sheep, cattle, fish, and birds. A human subject can refer to a patient.

[0094] The term "suspension" as used in this article refers to a liquid form of drug delivery that contains solid particles dispersed in a liquid carrier.

[0095] As used herein, “viscosity” refers to the resistance to fluid flow. Viscosities may be used herein, including, for example, polyvinyl alcohol, polyvinylpyrrolidone, methylcellulose, hydroxypropyl methylcellulose, hydroxyethylcellulose, carboxymethylcellulose, hydroxypropylcellulose, other reagents known to those skilled in the art, or combinations thereof.

[0096] The term "weight percentage" or "% (w / w)" refers to the percentage of a component in a solution based on the weight of the component and the solvent. For example, a 1% (w / w) solution of a component can dissolve 1 g of the component in 100 g of solvent. The term "volume percentage" or "% (v / v)" refers to the percentage of a component in a solution based on the volume of the component and the solvent. For example, a 1% (v / v) solution of a component can dissolve 1 ml of the component in 100 ml of solvent. The term "weight / volume percentage" or "% (w / v)" refers to the percentage of a component in a solution based on the weight of the component and the volume of the solvent. For example, a 1.0% (w / v) solution of a component can have 1 g of the component dissolved in 100 ml of solvent.

[0097] As used in this article, the term "syndrome" refers to a condition characterized by a group of symptoms or a series of related symptoms that occur consecutively. A syndrome (e.g., acute respiratory distress syndrome) may be a set of interconnected medical signs and symptoms that are usually associated with a specific disease. On the other hand, a disease may be a health condition with a clearly defined cause. However, a syndrome (from the Greek word meaning "running together") may cause many symptoms without a clear cause. These may suggest the possibility of an underlying disease or the likelihood of disease progression.

[0098] The terms “treat,” “treating,” or “treatment,” and other grammatical equivalents as used herein, include the reduction, attenuation, improvement, or prevention of a disease, condition (e.g., acute respiratory distress syndrome), or symptoms; prevention of additional symptoms; improvement or prevention of the underlying metabolic cause of symptoms; inhibition of a disease or condition, such as stagnation of its development; remission, extinction, or cessation of symptoms caused by the disease or condition; and are intended to include prevention. The term further includes achieving therapeutic and / or preventive benefits. A therapeutic benefit means the eradication or improvement of the underlying disease being treated. Furthermore, because a therapeutic benefit is achieved by eradicating or improving one or more physiological symptoms associated with the underlying condition, improvement is observed in the patient even if the patient may still have the underlying condition.

[0099] Health functional foods refer to foods or food supplements prepared or processed from raw materials, functional ingredients, active pharmaceutical ingredients or additives, and used to improve and / or nourish and / or maintain human physiological functions.

[0100] The term "acute respiratory distress syndrome (ARDS)" refers to a medical condition in severely ill patients with widespread inflammation. ARDS is a clinical phenotype that can be caused by a variety of pathologies, including pneumonia and sepsis. ARDS is characterized by the activation of the innate immune response resulting from alveolar barrier dysfunction, surfactant dysfunction, abnormal coagulation, and widespread cellular damage.

[0101] The term "acute lung injury (ALI)" refers to an inflammatory syndrome and increased permeability associated with hypoxemia and a classic radiographic appearance. At the most severe end of this range is ARDS.

[0102] The term "airway surface fluid (ASL)" refers to a thin layer of fluid covering the apical surface of the airway epithelium. ASL plays a crucial role in maintaining airway homeostasis. ASL volume, pH, and ion balance are directly involved in the regulation of antimicrobial activity, ciliary function, and mucosal clearance.

[0103] The term "inflammatory airway disease" refers to various inflammatory airway diseases, including 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), chronic obstructive pulmonary disease (COPD), etc.

[0104] In the context of this 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 one or more targets or two or more targets that induce a desired biological effect. Non-limiting examples of targets include enzymes, receptors, ion channels, or transport proteins (e.g., pantelin). An "inhibitor" can reversibly or irreversibly inhibit a target; reversible inhibition includes competitive inhibition, non-competitive inhibition, non-competitive inhibition, and mixed inhibition. The term "alkyl" includes straight-chain or branched C1-C... 20 Alkyl, when used alone or in conjunction with other terms, refers to a monovalent alkyl group having 1 to 20 carbon atoms. Examples of this term include groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-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-pentadecanyl, n-hexadecyl, n-octadecyl, n-nonadecanyl, and n-eicosyl. Preferably, this includes C1-C9 alkyl, more preferably C1-C6 alkyl, particularly preferably C1-C4 alkyl, and similarly, represents a monovalent alkyl having 1 to 9 carbon atoms, specifically a monovalent alkyl having 1 to 6 carbon atoms and a monovalent alkyl having 1 to 4 carbon atoms.

[0105] The term "alkenyl" includes straight-chain or branched C2-C chains. 20Alkenyl, when used alone or in conjunction with other terms. It can have any number of double bonds in any available position, and the configuration of the double bonds can be (E) or (Z) configuration. Examples of this term include vinyl, allyl, isopropylphenyl, 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-eicosenoenyl, and 3,7,11,15-tetramethyl-1-hexadecenyl, etc. Preferably, this includes C2-C8 alkenyl groups, more preferably C2-C6 alkenyl groups. Among these, vinyl or vinyl (-CH=CH2), n-2-propenyl (allyl, -CH2CH=CH2), particularly preferably isopropylphenyl, 1-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, 3-methyl-2-butenyl, etc.

[0106] The term "alkynyl" includes straight-chain or branched C2-C groups. 20 The term alkynyl, when used alone or in conjunction with other terms, can have any number of triple bonds in any available position. Examples of this term include groups such as alkynyl groups that can have 2 to 20 carbon atoms and any double or triple bonds, such as ethynyl (-C≡1-propynyl, 2-propynyl (propynyl: -CH2C≡2-butynyl, 2-penten-4-alkynyl), etc. In particular, this includes C2-C8 alkynyl groups, more preferably, C2-C6 alkynyl groups, etc. Preferably, it includes C2-C6 alkynyl groups, referring to groups having 2 to 6 carbon atoms and having alkynyl unsaturation in at least one or two positions.

[0107] The term "heteroalkyl" refers to C1-C 12 -alkyl, preferably C1-C6-alkyl, and at least one carbon atom is substituted with a heteroatom selected from O, N or S, including 2-methoxyethyl, etc.

[0108] The term "aryl" refers to an unsaturated aromatic carbocyclic group (e.g., indenyl, naphthyl, 2,3-dihydro-1H-indenyl, 1,2,3,4-tetrahydronaphthyl) with 6 to 14 carbon atoms, having a monocyclic (e.g., phenyl) or polycyclic rings. Aryl groups include phenyl, naphthyl, anthracene, phenanthrene, etc.

[0109] The term "C1-C6 alkyl aryl" refers to aryl groups having C1-C6 alkyl substituents, including methylphenyl, ethylphenyl, tert-butylphenyl, etc.

[0110] The term "aryl C1-C6 alkyl" refers to C1-C6 alkyl groups with aryl substituents, including 3-phenylpropyl, benzyl, etc.

[0111] The term "heteroaryl" refers to a monocyclic, bicyclic, or tricyclic fused-ring heteroaryl group. Specific examples of heteroaryl groups include optionally substituted pyridyl, pyrroloyl, pyrimidinyl, furanyl, thiophenyl, imidazolyl, etc. azole group, iso Azolyl, thiazolyl, isothiazolyl, pyrazolyl, 1H-pyrazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3- Diazolyl, 1,2,4- Diazolyl, 1,2,5- Diazolyl, 1,3,4- Diazolyl, 1,3,4-triazinyl, 1,2,3-triazinyl, benzofuranyl, [2,3-dihydro]benzofuranyl, isobenzofuranyl, benzothiophenyl, benzotriazolyl, isobenzothiophenyl, indoleyl, isoindoleyl, 3H-indoleyl, benzimidazoyl, imidazo[1,2-a]pyridyl, benzothiazoyl, benzo[…] Azolyl, quinolinyl, quinazolinyl, phthalazinyl, quinoxalinyl, cinnamyl, naphridyl, pyrido[3,4-b]pyridyl, pyrido[3,2-b]pyridyl, pyrido[4,3-b]pyridyl, quinolinyl, isoquinolinyl, tetrazolyl, 5,6,7,8-tetrahydroquinolinyl, 5,6,7,8-tetrahydroisoquinolinyl, purinyl, pteridyl, carbazoleyl, xanthineyl, benzoquinolinyl, benzo[d][1,3]dioxolane-5-yl, 3,4-dihydro-1H-pyrano[4,3-c]pyridyl, quinolin-2(1H)-one, 4H-chromene, 1H-indole, etc.

[0112] The term "C1-C6 alkyl heteroaryl" refers to heteroaryl groups with C1-C6 alkyl substituents, including methylfuranyl, tert-butylfuranyl, etc.

[0113] The term "heteroaryl C1-C6 alkyl" refers to C1-C6 alkyl groups having heteroaryl substituents, including furanylmethyl, etc.

[0114] The term "C2-C6 alkenylaryl" refers to an aryl group having C2-C6 alkenyl substituents, including vinylphenyl, etc.

[0115] The term "aryl C2-C6 alkenyl" refers to C2-C6 alkenyl groups with aryl substituents, including phenyl vinyl groups, etc.

[0116] The term "C2-C6 alkenyl heteroaryl" refers to heteroaryl groups with C2-C6 alkenyl substituents, including vinylpyridyl, etc.

[0117] The term "heteroaryl C2-C6 alkenyl" refers to C1-C6 alkenyl groups with heteroaryl substituents, including pyridyl vinyls, etc.

[0118] The term "C3-C8-cycloalkyl" refers to a saturated carbocyclic group having 3 to 8 carbon atoms, either a monocyclic (e.g., cyclohexyl) or polycyclic (e.g., norbornyl). Cycloalkyl groups include cyclopentyl, cyclohexyl, norbornyl, etc.

[0119] The term "heterocyclic alkyl" refers to a C3-C8 cycloalkyl or polyfused ring, wherein up to three carbon atoms are substituted with heteroatoms selected from the group consisting of O, S, and NR (R is defined as hydrogen or methyl). Heterocyclic alkyl groups include lactams or lactones. Non-limiting examples are pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, decahydroisoquinolinyl, octahydro-1H-pyrano[3,4-c]pyridinyl, 4-methylene-5(4H)-one, pyrrolidine-2-one, etc.

[0120] The term "C1-C6 alkyl-C3-C8 cycloalkyl" refers to C3-C8 cycloalkyl groups having C1-C6 alkyl substituents, including methylcyclopentyl, etc.

[0121] The term "C3-C8-cycloalkylC1-C6 alkyl" refers to C1-C6 alkyl groups having C3-C8-cycloalkyl substituents, including 3-cyclopentylpropyl, etc.

[0122] The term "C1-C6 alkyl heterocyclic alkyl" refers to heterocyclic alkyl groups having C1-C6 alkyl substituents, including 4-methylpiperidinyl, etc.

[0123] The term "heterocyclic alkyl C1-C6 alkyl" refers to C1-C6 alkyl groups having heterocyclic alkyl substituents, including (1-methylpiperidin-4-yl)methyl, etc.

[0124] The term "carboxyl group" refers to the group -C(O)OH.

[0125] The term "carboxylated C1-C6 alkyl" refers to C1-C6 alkyl groups having carboxyl substituents, including 2-carboxyethyl, etc.

[0126] The term "acyl" refers to the group -C(O)R, including acetyl, etc., where R includes H, "alkyl" is preferably "C1-C6 alkyl", "aryl", "heteroaryl", "C3-C8 cycloalkyl", "heterocyclic alkyl", "aryl C1-C6 alkyl", "heteroaryl C1-C6 alkyl", "C3-C8 cycloalkyl C1-C6 alkyl" or "heterocyclic alkyl C1-C6 alkyl".

[0127] The term "acyl C1-C6 alkyl" refers to C1-C6 alkyl groups having acyl substituents, including 2-acetylacetyl, etc.

[0128] The term "acylaryl" refers to an aryl group having an acyl substituent, including 2-acetylphenyl, etc.

[0129] The term "acyloxy group" refers to the group -OC(O)R, including acetoxy, etc., and R includes H, "C1-C6 alkyl", "C2-C6 alkenyl", "C2-C6 ynyl", "C3-C8-cycloalkyl", "heterocyclic alkyl", "aryl", "heteroaryl", "aryl C1-C6 alkyl", "heteroaryl C1-C6 alkyl", "aryl C2-C6 alkenyl", "heteroaryl C2-C6 alkenyl", "aryl C2-C6 ynyl", "heteroaryl C2-C6 ynyl", "C3-C8-cycloalkyl C1-C6 alkyl" or "heterocyclic alkyl C1-C6 alkyl".

[0130] The term "acyloxy C1-C6 alkyl" refers to C1-C6 alkyl groups having acyloxy substituents, including 2-(ethylcarbonyloxy)ethyl, etc.

[0131] The term "alkoxy" refers to the group -OR, where R includes "C1-C6 alkyl", "aryl", "heteroaryl", "aryl C1-C6 alkyl", or "heteroaryl C1-C6 alkyl". Preferred alkoxy groups include, for example, methoxy, ethoxy, phenoxy, etc.

[0132] The term "alkoxy C1-C6 alkyl" refers to C1-C6 alkyl groups having alkoxy substituents, including methoxyethyl, etc.

[0133] The term "alkoxycarbonyl" refers to the group -C(O)OR, where R includes "C1-C6 alkyl", "aryl", "heteroaryl", "arylC1-C6 alkyl", "heteroarylC1-C6 alkyl" or "heteroalkyl".

[0134] The term "alkoxycarbonyl C1-C6 alkyl" refers to C1-C6 alkyl groups having alkoxycarbonyl substituents, including 2-(benzyloxycarbonyl)ethyl, etc.

[0135] The term "aminocarbonyl" refers to the group -C(O)NRR', including N-phenylcarbonyl, etc., where R and R' are independently H, C1-C6 alkyl, aryl, heteroaryl, "aryl C1-C6 alkyl" or "heteroaryl C1-C6 alkyl".

[0136] The term "aminocarbonyl C1-C6 alkyl" refers to an alkyl group having an aminocarbonyl substituent, including 2-(dimethylaminocarbonyl)ethyl, N-ethylacetamido, N,N-diethylacetamido, etc.

[0137] The term "acylamino" refers to the group -NRC(O)R', including acetamino, etc., where R and R' are independently H, and it is "C1-C6 alkyl", "C2-C6 alkenyl", "C2-C6 ynyl", "C3-C8-cycloalkyl", "heterocycloalkyl", "aryl", "heteroaryl", "aryl C1-C6 alkyl", "heteroaryl C1-C6 alkyl", "aryl C2-C6 alkenyl", "heteroaryl C2-C6 alkenyl", "aryl C2-C6 ynyl", "heteroaryl C2-C6 ynyl", "cycloalkyl C1-C6 alkyl" or "heterocycloalkyl C1-C6 alkyl".

[0138] The term "acylamino C1-C6 alkyl" refers to C1-C6 alkyl groups having acylamino substituents, including 2-(propionylamino)ethyl, etc.

[0139] The term "ureido" refers to the group -NRC(O)NR'R", where R, R', and R" are independently H, and can be "C1-C6 alkyl", "alkenyl", "alkynyl", "C3-C8 cycloalkyl", "heterocyclic alkyl", "C1-C6 aryl", "heteroaryl", "aryl C1-C6 alkyl", "heteroaryl C1-C6 alkyl", "aryl C2-C6 alkenyl", "heteroaryl C2-C6 alkenyl", "aryl C2-C6 alkynyl", "heteroaryl C2-C6 alkynyl", "cycloalkyl C1-C6 alkyl", or "heterocyclic alkyl C1-C6 alkyl". R' and R" may optionally form a 3-8 membered heterocyclic alkyl ring together with the nitrogen atom attached to them.

[0140] The term "ureido-C1-C6 alkyl" refers to C1-C6 alkyl groups having ureido substituents, including 2-(N'-methylureido)ethyl, etc.

[0141] The term "carbamate" refers to the group -NRC(O)OR', where R and R' are independently "C1-C6 alkyl", "C2-C6 alkenyl", "C2-C6 ynyl", "C3-C8-cycloalkyl", "heterocycloalkyl", "aryl", "heteroaryl", "C1-C6 alkyl aryl", "heteroaryl C1-C6 alkyl", "aryl C2-C6 alkenyl", "heteroaryl C2-C6 alkenyl", "aryl C2-C6 ynyl", "heteroaryl C2-C6 ynyl", "cycloalkyl C1-C6 alkyl", or "heterocycloalkyl C1-C6 alkyl", and R can be hydrogen.

[0142] The term "amino" refers to the group -NRR', and then R and R' are independently H, "C1-C6 alkyl", "aryl", "heteroaryl", "C1-C6 alkylaryl", "C1-C6 alkylheteroaryl", "cycloalkyl" or "heterocyclic alkyl", and then R and R' may optionally form a 3-8 membered heterocyclic alkyl ring together with the nitrogen atom attached to them.

[0143] The term "aminoalkyl" refers to an alkyl group having an amino substituent, including 2-(1-pyrrolidinyl)ethyl, etc.

[0144] The term "ammonium" refers to the positively charged group -N. + RR'R", then R, R' and R" are independently "C1-C6 alkyl", "C1-C6 alkylaryl", "C1-C6 alkylheteroaryl", "cycloalkyl" or "heterocyclic alkyl", then R and R' may optionally form a 3-8 membered heterocyclic alkyl ring with the nitrogen atom to which they are attached.

[0145] The term "ammonium alkyl" refers to an alkyl group having an ammonium substituent, including 1-ethylpyrrolidineonium, etc.

[0146] The term "halogen" refers to fluorine, chlorine, bromine, and iodine atoms.

[0147] The term "sulfonyloxy" refers to the group -OSO2R, where R is selected from "C1-C6 alkyl" or "C1-C6 alkyl" that has been substituted with a halogen, such as -OSO2CF3 group, "C2-C6 alkenyl", "alkynyl", "C3-C8 cycloalkyl", "heterocyclic alkyl", "aryl", "heteroaryl", "aryl C1-C6 alkyl", "heteroaryl C1-C6 alkyl", "aryl C2-C6 alkenyl", "heteroaryl C2-C6 alkenyl", "aryl C2-C6 alkynyl", "heteroaryl C2-C6 alkynyl", "cycloalkyl C1-C6 alkyl", or "heterocyclic alkyl alkyl".

[0148] The term "sulfonyloxy C1-C6 alkyl" refers to an alkyl group having a sulfonyloxy substituent, including 2-(methylsulfonyloxy)ethyl, etc.

[0149] The term "sulfonyl" refers to the group "-SO2R", where R is selected from "aryl", "heteroaryl", "C1-C6 alkyl", "C1-C6 alkyl" substituted with halogen, for example, -SO2CF3, "C2-C6 alkenyl", "C2-C6 ynyl", "C3-C8 cycloalkyl", "heterocycloalkyl", "aryl", "heteroaryl", "aryl C1-C6 alkyl", "heteroaryl C1-C6 alkyl", "aryl C2-C6 alkenyl", "heteroaryl C2-C6 alkenyl", "aryl C2-C6 ynyl", "heteroaryl C2-C6 ynyl", "cycloalkyl C1-C6 alkyl", or "heterocycloalkyl C1-C6 alkyl".

[0150] The term "sulfonyl C1-C6 alkyl" refers to an alkyl group having a sulfonyl substituent, including 2-(methylsulfonyl)ethyl, etc.

[0151] The term "sulfinyl" refers to the "-S(O)R" group, where R is selected from "alkyl", halogen-substituted "alkyl", such as -SOCF3 group, "C2-C6 alkenyl", "C2-C6 ynyl", "C3-C8 cycloalkyl", "heterocyclic alkyl", "aryl", "heteroaryl", "aryl C1-C6 alkyl", "heteroaryl C1-C6 alkyl", "aryl C2-C6 alkenyl", "heteroaryl C2-C6 alkenyl", "aryl C2-C6 ynyl", "heteroaryl C2-C6 ynyl", "C3-C8-cycloalkyl C1-C6 alkyl" or "heterocyclic alkyl C1-C6 alkyl".

[0152] The term "sulfinyl alkyl" refers to an alkyl group having a sulfinyl substituent, including 2-(methylsulfinyl)ethyl, etc.

[0153] The term "sulfanyl" refers to the group -SR, where R includes H, "C1-C6 alkyl", halogen-substituted "C1-C6 alkyl", such as -SCF3-yl, "C2-C6 alkenyl", "C2-C6 ynyl", "C3-C8-cycloalkyl", "heterocycloalkyl", "aryl", "heteroaryl", "aryl C1-C6 alkyl", "heteroaryl C1-C6 alkyl", "aryl C2-C6 alkenyl", "heteroaryl C2-C6 alkenyl", "aryl C2-C6 ynyl", "ynyl heteroaryl", "cycloalkyl C1-C6 alkyl", or "heterocycloalkyl C1-C6 alkyl". Preferred sulfanyl groups include methyl sulfanyl, ethyl sulfanyl, etc.

[0154] The term "thioalkyl C1-C6 alkyl" refers to C1-C5-alkyl groups having thioalkyl substituents, including 2-(ethylthioalkyl)ethyl, etc.

[0155] The term "sulfonamide" refers to the group -NRSO2R', where R and R' are independently "C1-C6 alkyl", "C2-C6 alkenyl", "C2-C6 alkynyl", "C3-C8-cycloalkyl", "heterocycloalkyl", "aryl", "heteroaryl", "aryl C1-C6 alkyl", "heteroaryl C1-C6 alkyl", "aryl C2-C6 alkenyl", "heteroaryl C2-C6 alkenyl", "aryl C2-C6 alkynyl", "heteroaryl C2-C6 alkynyl", "C3-C8 cycloalkyl C1-C6 alkyl", or "heterocycloalkyl C1-C6 alkyl".

[0156] The term "sulfonamide C1-C6 alkyl" refers to an alkyl group having a sulfonamide substituent, including 2-(ethylsulfonamide)ethyl, etc.

[0157] The term "aminosulfonyl" refers to the group -SO2NRR', where R and R' are independently H, and can be "C1-C6 alkyl", "C2-C6 alkenyl", "C2-C6 ynyl", "C3-C8-cycloalkyl", "heterocyclic alkyl", "aryl", "heteroaryl", "aryl C1-C6 alkyl", "heteroaryl C1-C6 alkyl", "aryl alkenyl", "heteroaryl C2-C6 alkenyl", "aryl C2-C6 ynyl", "heteroaryl C2-C6 ynyl", "C3-C8-cycloalkyl C1-C6 alkyl", or "heterocyclic alkyl C1-C6 alkyl". Optionally, R and R' can form a 3-8 membered heterocyclic alkyl ring with the nitrogen atom to which they are attached. Aminosulfonyl groups include cyclohexylaminosulfonyl, piperidinylsulfonyl, etc.

[0158] The term "aminosulfonyl C1-C6 alkyl" refers to a C1-C6 alkyl group having an aminosulfonyl substituent, including 2-(cyclohexylaminosulfonyl)ethyl, etc. Unless otherwise defined, all substituents should be understood as being optionally substituted.

[0159] Unless otherwise defined by the specific substituent, the term "substituted" refers to a group substituted by one to five substituents selected from the group consisting of: "C1-C6 alkyl", "C2-C6 alkenyl", "C2-C6 alkynyl", "C3-C8 cycloalkyl", "heterocycloalkyl", "C1-C6 alkylaryl", "C1-C6 alkylheteroaryl", "C1-C6 alkylcycloalkyl", "C1-C6 alkylheterocycloalkyl", "amino", "aminosulfonyl", "ammonium", "acylamino", "aminocarbonyl", "aryl", "heteroaryl", "sulfinyl", "sulfonyl", "alkoxy", "alkoxycarbonyl", "carbamate", "thioalkyl", "halogen", trihalomethyl, cyano, hydroxyl, mercapto, nitro, etc."

[0160] 2. Compounds

[0161] This invention provides, in one aspect, compounds represented by the following chemical formula 1, their E- or Z-isomers, their optical isomers, mixtures of the two isomers, their precursors, their pharmaceutically acceptable salts, or their solvates:

[0162] Chemical Formula 1

[0163]

[0164] In chemical formula 1,

[0165] V 1 and V 2 It is aryl, heteroaryl, C3-C7 cycloalkyl, heterocycloalkyl, C1-C6 alkyl, C1-C6 heteroalkyl, C2-C 10alkenyl, C0-C3 methylene hydrazine, C2-C 10 Alkyne group, S(O) i (C1-C6 alkyl), OS(O) i (Aryl), S(O) i NR 3 R 4 C(O)R 3 OR 3 OCOR 3 NR 3 C(O)OR 4 NR 3 C(O)R 4 C(O)NR 3 R 4 Or NR 3 R 4 Furthermore, aryl, heteroaryl, C3-C7 cycloalkyl, heterocycloalkyl, C1-C6 alkyl, C1-C6 heteroalkyl, C2-C 10 alkenyl, C0-C3 methylene hydrazine, C2-C 10 Alkyne group, S(O) i (C1-C6 alkyl), OS(O) i (Aryl), S(O) i NR 3 R 4 C(O)R 3 OR 3 OCOR 3 NR 3 C(O)OR 4 NR 3 C(O)R 4 C(O)NR 3 R 4 and NR 3 R 4 Optionally, one or more of the groups are independently selected from hydrogen, oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, sulfonyl, aryl, C1-C 10 Alkyl, C3-C7 cycloalkyl, heteroaryl, heterocycloalkyl, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i NR 3 (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 OCR 3 F2, OCOR3 NR 3 C(O)OR 4 NR 3 C(O)R 4 C(O)NR 3 R 4 and NR 3 R 4 The groups are substituted, and the aryl, C1-C1 groups are substituted. 10 Alkyl, C3-C7 cycloalkyl, heteroaryl, heterocycloalkyl, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i NR 3 (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 OCR 3 F2, OCOR 3 NR 3 C(O)OR 4 NR 3 C(O)R 4 C(O)NR 3 R 4 and NR 3 R 4 Optionally, one or more of the radicals are independently selected from hydrogen, oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, aryl, C1-C2. 10 Alkyl, aryl, C3-C7 cycloalkyl, heteroaryl, heterocycloalkyl, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i (Aryl), S(O) i (Heteroaryl), S(O) i NR 3 (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 OCR 3 F2, OCOR 3 NR 3 C(O)OR 4 NR 3 C(O)R 4 C(O)NR3 R 4 and NR 3 R 4 Group substitution, and

[0166] i and j are independently 0, 1, or 2, and

[0167] R 1 and R 2 Independently selected from hydrogen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, sulfonyl, aryl, C1-C 10 Alkyl, heteroaryl, heterocyclic, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 The group consists of aryl, heteroaryl, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 Optionally, one or more of the groups are independently selected from oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, aryl, heteroaryl, heterocyclic, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R4 and NR 3 R 4 Group substitution, and

[0168] R 3 and R 4 Independently selected from hydrogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, sulfonyl, aryl, aryl (C1-C2) 10 Alkyl groups, C1-C6 10 Alkyl, heteroaryl, heteroaryl (C1-C2) 10 Alkyl groups, C1-C6 10 Alkyl heteroaryl, C1-C 10 The group consisting of alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, heterocyclic and trifluoromethyl groups, and aryl, aryl (C1-C6) 10 Alkyl groups, C1-C6 10 Alkyl, heteroaryl, heteroaryl (C1-C2) 10 Alkyl groups, C1-C6 10 Alkyl heteroaryl, C1-C 10 Alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, and heterocyclic groups are optionally selected independently by one or more groups selected from hydrogen, oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, aryl, heteroaryl, heterocyclic, C1-C6 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 Substitution of groups, or

[0169] R 3 and R 4 It can be cyclized into a 4- to 10-membered carbocyclic, heterocyclic, aromatic, or heteroaromatic ring, and one of the carbocyclic, heterocyclic, aromatic, or heteroaromatic rings may optionally be selected independently from one or more elements, including hydrogen, oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, aryl, C1-C2. 10 Alkyl, heteroaryl, heterocyclic, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 Group substitution, and

[0170] A 1 Represented as or and

[0171] X 1 and X 2 Independently select O, S, CHR 4 and NR 4 The group formed, and

[0172] X 3 Choose free hydrogen, OR 3 aryl, heteroaryl, C3-C7 cycloalkyl, heterocycloalkyl, C1-C6 alkyl, C1-C6 heteroalkyl, C2-C 10 alkenyl, C0-C3 methylene hydrazine, C2-C 10 Alkyne group, S(O) i (C1-C6 alkyl), S(O) i NR 3 C(O)R 3 OC(O)R 3 (O)COR 3 NR 3 C(O)OR 3 NR 3 C(O)R 3 C(O)NR 3 and NR 3 R 4 The group formed, and OR 3 aryl, heteroaryl, C3-C7 cycloalkyl, heterocycloalkyl, C1-C6 alkyl, C1-C6 heteroalkyl, C2-C 10 alkenyl, C0-C3 methylene hydrazine, C2-C 10 Alkyne group, S(O) i (C1-C6 alkyl), S(O) i NR 3 C(O)R 3 OC(O)R 3 (O)COR 3 NR3 C(O)OR 3 NR 3 C(O)R 3 C(O)NR 3 and NR 3 R 4 One of them is optionally selected independently from hydrogen, oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, sulfonyl, aryl, C1-C 10 Alkyl, C3-C7 cycloalkyl, heteroaryl, heterocycloalkyl, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i NR 3 (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 OCR 3 F2, OCOR 3 NR 3 C(O)OR 4 NR 3 C(O)R 4 C(O)NR 3 R 4 and NR 3 R 4 The groups in, and aryl, C1-C 10 Alkyl, C3-C7 cycloalkyl, heteroaryl, heterocycloalkyl, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i NR 3 (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 OCR 3 F2, OCOR 3 NR 3 C(O)OR 4 NR 3 C(O)R 4 C(O)NR 3 R 4 and NR 3 R 4Optionally, one or more of the groups are independently selected from hydrogen, oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, sulfonyl, aryl, C1-C 10 Alkyl, aryl, C3-C7 cycloalkyl, heteroaryl, heterocycloalkyl, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i (Aryl)), S(O) i (Heteroaryl), S(O) i NR 3 (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 OCR 3 F2, OCOR 3 NR 3 C(O)OR 4 NR 3 C(O)R 4 C(O)NR 3 R 4 and NR 3 R 4 Group substitution, and

[0173] R 5 and R 6 Independently selected from hydrogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, sulfonyl, aryl, C1-C 10 Alkyl, heteroaryl, heterocyclic, C1-C 10 The group consisting of alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, heterocyclic, aryl, heteroaryl, and trifluoromethyl, wherein alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, C1-C6... 10 One of the alkylaryl and heteroaryl groups is optionally selected from one or more groups, and is independently selected from hydrogen, oxygen, halogen, cyano, azathiol, nitro, trifluoromethyl, trifluoromethoxy, aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclic, C1-C2. 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i (Aryl), S(O) i (Heteroaryl), S(O) i NR 3 R 4C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 One or more groups in the group, and aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclic, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i (Aryl), S(O) i (Heteroaryl), S(O) i NR 3 R 4 C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 One of the groups is optionally selected from one or more independently selected from hydrogen, oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclic, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1-C6 alkyl), S(O) i (Aryl), S(O) i (Heteroaryl), S(O) i NR 3 R 4 C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 One or more groups in, or

[0174] R 5 and A1 It can be cyclized into a 4- to 10-membered carbocyclic, heterocyclic, aromatic, or heteroaromatic ring, and one of the carbocyclic, heterocyclic, aromatic, or heteroaromatic rings may optionally be selected independently from one or more elements, including oxygen, halogen, cyano, azathio, nitro, trifluoromethyl, trifluoromethoxy, aryl, C1-C2, etc. 10 Alkyl, heteroaryl, heterocyclic, C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne group, C3-C6 cycloalkyl group, S(O) i (C1~C6 alkyl), C(O)OR 3 C(O)R 3 OR 3 NR 3 C(O)OR 4 C(O)NR 3 R 4 and NR 3 R 4 Substitution of groups in [the group].

[0175] Non-limiting examples of compounds include the following compounds described in Tables 1 and 2.

[0176] [Table 1]

[0177]

[0178]

[0179]

[0180]

[0181]

[0182]

[0183]

[0184]

[0185]

[0186] [Table 2]

[0187]

[0188]

[0189]

[0190] In this document, the term "compound of the present invention" and equivalent expressions include the compounds represented by Formula 1 above, and such expressions include their E- or Z-isomers, their optical isomers, mixtures of the two isomers, their precursors, their pharmaceutically acceptable salts or solvates thereof, and are newly synthesized.

[0191] The invention is further described by the following examples, unless they limit the scope of the invention.

[0192] 3. Preparation method

[0193] Another aspect of the present invention provides a method for preparing a compound represented by chemical formula 1. The synthetic method is described in detail in the detailed embodiments described herein.

[0194] In some embodiments, the base used during organic synthesis can be an organic or inorganic base. Non-limiting examples of organic bases include pyridine, trimethylamine, N,N-diisopropylethylamine (DIPEA), and 1,8-diazabicyclo[5,4,0]undec-7-ene (DBU). Non-limiting examples of inorganic bases include sodium hydroxide, sodium carbonate, potassium carbonate, cesium carbonate, and sodium hydride. They can be used alone or in stoichiometric or excess combinations. 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. Solvents can be used alone or in combination.

[0195] 4. Composition / Formulation

[0196] This invention includes the compounds described herein and pharmaceutical compositions comprising formulations suitable for administering the compounds described herein. It includes formulations of pharmaceutical compositions suitable for administration by any medically acceptable manner. Pharmaceutical formulations may comprise pharmaceutically acceptable additives or carriers suitable for the manner of administration and pharmaceutically acceptable compounds (compositions).

[0197] The compounds described herein can be formulations (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), isotonic adjusters (e.g., one or more isotonic adjusters), thickeners (e.g., one or more thickeners), suspending agents (e.g., one or more suspending agents), binders (e.g., one or more binders), viscosity increasers (e.g., one or more viscosity increasers), etc., and are provided as pharmaceutically acceptable adjuncts for a specific condition to be treated. In some embodiments, the formulation may contain a combination of the adjuncts described herein (e.g., 2, 3, 4, 5, 6, 7, 8 or more adjuncts). In some embodiments, the additive may include, for example, therapeutic agents and drug delivery modifiers, as well as reinforcing agents such as calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starch, gelatin, cellulose, methylcellulose, sodium carboxymethylcellulose, glucose, hydroxypropyl-β-cyclodextrin, polyvinylpyrrolidone, low-melting-point waxes and ion exchange resins, and combinations of any two or more of these.

[0198] 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 (2003) and 21st (2005), respectively, which are incorporated herein by reference.

[0199] The formulations of the compositions described herein are suitable for oral administration and may be in the form of inhalation, nasal spray, intravenous, intramuscular injection, intravitreal injection, ointment or solution, suspension, semi-liquid, semi-solid, gel, semi-solid gel, jelly, emulsion, ointment, tablet, liquid, and paste. Tablet forms may contain one or more of lactose, sucrose, mannitol, sorbitol, calcium phosphate, corn starch, potato starch, microcrystalline cellulose, gelatin, colloidal silica, talc, magnesium stearate, stearic acid, and other excipients, colorants, fillers, binders, diluents, buffers, humectants, preservatives, flavoring agents, dyes, disintegrants, and pharmaceutically suitable carriers. Capsules may contain suitable excipients and compounds, or compounds that can be used alone in the shell. All these formulated compounds may be administered alone or in combination, intermittently, sequentially, or simultaneously.

[0200] 5. Administration

[0201] The compositions of the present invention comprise, but are not limited to, compositions that can be administered orally, parenterally, sublingually, dermally, rectally, transmucosally, topically, by inhalation, buccally or intranasally, or by any combination thereof. Parenterally administration includes, but is not limited to, intravenous, intra-arterial, intraperitoneal, subcutaneous, intramuscular, intrathecal, and intra-arterial administration. Furthermore, the compositions of the present invention can be administered as implants, which allows for slow release of the composition and slow, controlled intravenous administration.

[0202] The dosage given to an individual, whether as a single dose or multiple doses, will vary depending on a variety of factors, including pharmacokinetic characteristics, patient condition and characteristics (sex, age, weight, health, body type), severity of symptoms, concurrent treatment, frequency of treatment, and expected outcome.

[0203] According to one embodiment of the present invention, the compounds and pharmaceutical preparations thereof according to the present invention can be administered alone or in combination with a useful adjuvant for the treatment of respiratory disorders or diseases. According to another embodiment of the present invention, the compounds and pharmaceutical preparations thereof according to the present invention can be administered in combination with radiotherapy.

[0204] This invention includes administering a compound or pharmaceutical preparation according to the invention to a subject simultaneously or sequentially at a therapeutically effective amount before other therapies or adjuvants (e.g., combination therapies) for treating cancer. The compound or pharmaceutical preparation according to the invention administered with an adjuvant may be administered in the same or different compositions via the same or different routes of administration.

[0205] In one embodiment, the patient according to the invention may be a patient suffering from respiratory disorders or diseases such as bronchial asthma, bronchitis, allergic rhinitis, adult respiratory syndrome, cystic fibrosis, pulmonary viral infection (influenza), pulmonary hypertension, idiopathic pulmonary fibrosis, and chronic obstructive pulmonary disease (COPD).

[0206] 6. Applications according to the invention

[0207] In other embodiments, the present invention provides the use of a compound represented by chemical formula 1, a mixture of the compounds, or a pharmaceutical composition thereof for the prevention, improvement, or treatment of respiratory diseases (inflammatory airway diseases).

[0208] In other embodiments, the present invention provides the use of a compound represented by Chemical Formula 1 and pharmaceutical compositions thereof as a panterine inhibitor.

[0209] In other embodiments, the use of a compound represented by Chemical Formula 1 and a pharmaceutical composition thereof is provided, which preserves the volume of airway surface fluid (ASL) and reduces mucin secretion.

[0210] In other embodiments, the present invention provides a 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 infection, cystic fibrosis, acute respiratory distress syndrome (ARDS), acute lung injury (ALI), or chronic obstructive pulmonary disease (COPD).

[0211] In other embodiments, the present invention provides the use of a compound represented by chemical formula 1 and pharmaceutical compositions thereof, wherein the respiratory disease (inflammatory airway disease) is selected from one or more of the group consisting of asthma, acute or chronic bronchitis, allergic rhinitis, acute respiratory infection, cystic fibrosis, acute respiratory distress syndrome (ARDS), acute lung injury (ALI), or chronic obstructive pulmonary disease (COPD).

[0212] Example

[0213] This document describes non-limiting embodiments of the experiment in detail, without limiting the scope of the experiment. The description of the invention herein is merely illustrative, and those skilled in the art should understand that it can be readily modified to other specific areas or forms without altering the technical spirit or essential characteristics of the invention. The illustrative description of the invention presents the following embodiments, but is not limited thereto.

[0214] Compound names are generated using ChemDraw Professional V.15.1. Compounds according to the invention include compounds represented by Formula 1, their tautomers, their geometric isomers (e.g., e, z isomers), their optically active forms as optical isomers, their diastereomers and their racemic mixtures, and their pharmaceutically acceptable salts. The derivatives exemplified herein 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 amounts of reagents, solvents, etc.), it should be understood that other experimental conditions may be used unless otherwise stated. Optimal reaction conditions may vary depending on the specific reactants or solvents, but such conditions can be determined by those skilled in the art using conventional optimization procedures.

[0215] The references cited herein are incorporated herein by reference in their entirety. This invention is not limited to the specific embodiments described herein, which are intended as individual examples of various aspects of the invention, and functionally equivalent methods and components are within the scope of the invention. In practice, various modifications to the invention will become apparent to those skilled in the art from the foregoing description and drawings, in addition to those shown and described herein. Such modifications are intended to fall within the scope of the appended claims.

[0216] Examples 1-2: Preparation of Novel Compounds

[0217] Due to the nature of synthetic chemical components or their inherent physicochemical properties, some synthetic compounds may exist as isomers or mixtures of isomers. In some cases, they may interconvert to other isomers. For example, compound 'F1' represents (E)-4-(thiophene-2-ylmethylene)-2-(4-(trifluoromethyl)phenyl) Azolium-5(4H)-one, (Z)-4-(thien-2-ylmethylene)-2-(4-(trifluoromethyl)phenyl) Azolium-5(4H)-one or a mixture of the two isomers. The study by Graziano et al. (Tetrahedron 62 (2006) 1165-1170) fully demonstrates this phenomenon.

[0218] Example 1.1: 4-(thiophene-2-ylmethylene)-2-(4-(trifluoromethyl)phenyl) Azolium-5(4H)-one (F1)

[0219]

[0220] Glycine (180 mg, 2.40 mmol) and 4-(trifluoromethyl)benzoyl chloride (0.411 mL, 2.758 mmol) were added sequentially to 24 mL of 10% NaOH solution under stirring at room temperature, and the mixture was stirred for 2 hours at the same temperature. After acidification to pH 2 with aqueous HCl, the resulting solid was filtered through a funnel and washed with chloride to obtain 2-(4-(trifluoromethyl)benzoylamino)acetate. This 2-(4-(trifluoromethyl)benzoylamino)acetate (529 mg, 2.14 mmol) and 1-ethyl-3-(3-dimethylamino)propylcarbodiimide hydrochloride (410 mg, 2.14 mmol) were added sequentially to 21 mL of dichloromethane under stirring at room temperature. Then, 1H-pyrrolo-2-carboxaldehyde (200 mg, 1.783 mmol) and triethylamine (0.497 mL, 3.567 mmol) were added with stirring at room temperature, and the mixture was stirred for 12 hours at this temperature. The solvent was removed by evaporation under reduced pressure. The resulting solid was filtered through a filter funnel and washed with methanol to provide the desired product (4-(thiophene-2-ylmethylene)-2-(4-(trifluoromethyl)phenyl) Azolium-5(4H)-one, F1).

[0221] 1 H NMR (400MHz, DMSO-d6) δ 8.22 (d, 2H, J = 8.4Hz), 8.08 (d, 1H, J = 5.2Hz), 7.96 (d, 2H, J = 8.4Hz), 7.86 (d, 1H, J = 3.6Hz), 7.79 (s, 1H), 7.24 (t, 1H, J = 4.2Hz).

[0222] ESI(m / z) 324(MH) + ).

[0223] Example 1.2: 2-(4-(tert-butyl)phenyl)-4-(thiazolyl-5-ylmethylene) Azolium-5(4H)-one (F2)

[0224]

[0225] Thionyl chloride (2.038 mL, 28.05 mmol) and dimethylformamide (10 drops) were added sequentially to 4-tert-butylbenzoate (500 mg, 2.81 mmol) in dichloromethane under stirring, and the mixture was heated for 12 hours. The solvent was evaporated to give an intermediate (0.542 mL, 2.758 mmol). Glycine (180 mg, 2.398 mmol) and the intermediate were added sequentially to a 10% NaOH aqueous solution (24 mL) under stirring at room temperature, and the mixture was stirred for 2 hours at this temperature. After acidification to pH 2 with aqueous HCl, the resulting solid was filtered through a funnel and washed with dichloromethane to give 2-(4-(tert-butyl)benzoylamino)acetate. 2-(4-(tert-butyl)benzoylamino)acetate (500 mg, 2.125 mmol) and 1-ethyl-3-(3-dimethylamino)propylcarbodiimide hydrochloride (448 mg, 2.338 mmol) were added sequentially to dichloromethane (21 mL) at room temperature. Thiazol-5-carboxaldehyde (216 mg, 1.913 mmol) and triacetamide (0.519 mL, 3.70 mmol) were added sequentially to the mixture at room temperature with stirring, and the mixture was stirred for 12 hours. The solvent was removed under reduced pressure. The solid was obtained by filtration through a funnel and washed with methanol to give the desired product (2-(4-(tert-butyl)phenyl)-4-(thiazol-5-ylmethylene)). Azolium-5(4H)-one, F2).

[0226] 1 H NMR (400MHz, DMSO-d6) δ 8.15 (d, 1H, J = 2.8Hz), 8.10 (d, 1H, J = 2.8Hz), 8.03 (d, 2H, J = 8.4Hz), 7.65 (d, 2H, J = 8.4Hz), 7.40 (s, 1H), 1.30 (s, 9H).

[0227] ESI(m / z) 313(MH) + ).

[0228] Example 1.3: 2-(4-(tert-butyl)phenyl)-4-((2-methyl-1H-indol-3-yl)methylene) Azoxyl-5(4H)-one (F3)

[0229]

[0230] Thionyl chloride (2.04 mL, 28.05 mmol) and dimethylformamide (10 drops) were added sequentially to 4-tert-butylbenzoate (500 mg, 2.81 mmol) in dichloromethane under stirring, and the mixture was heated for 12 hours. The solvent was evaporated to provide an intermediate (0.542 mL, 2.758 mmol). Glycine (180 mg, 2.398 mmol) and the intermediate were added sequentially to a 10% NaOH aqueous solution (24 mL) under stirring at room temperature. The entire mixture was stirred at room temperature for 2 hours. After acidification to pH 2 with HCl, the resulting solid was obtained through a filter funnel and washed with dichloromethane to provide 2-(4-(tert-butyl)benzoylamino)acetate. 2-(4-(tert-butyl)benzoylamino)acetate (400 mg, 2.073 mmol) and 1-ethyl-3-(3-dimethylamino)propylcarbodiimide hydrochloride (433.54 mg, 2.262 mmol) were added sequentially to dichloromethane (20 mL) with stirring at room temperature. 2-Methyl-1H-indole-3-carboxaldehyde (300 mg, 1.885 mmol) and triacetamide (0.315 mL, 2.262 mmol) were added sequentially to the mixture with stirring at room temperature, and the mixture was stirred for 12 hours. The solvent was removed under reduced pressure, and the solid was obtained by filtration through a funnel. The solid was washed with methanol to provide the desired product (2-(4-(tert-butyl)phenyl)-4-((2-methyl-1H-indole-3-yl)methylene). Azazole-5(4H)-one, F3).

[0231] 1 H NMR (400MHz, DMSO-d6) δ12.25 (s, 1H), 9.16 (d, 1H, J = 7.9Hz), 7.96 (d, 2H, J = 8.4Hz) ,7.61(d,2H,J=8.0Hz),7.37(s,2H),7.25-7.18(m,2H),2.62(s,3H),1.30(s,9H).

[0232] ESI (m / z) 359 (MH) + ),357(MH - ).

[0233] Example 1.4: 3-((2-(4-(tert-butyl)phenyl)-5-oxo tert-butyl azole-4(5H)-methylene)-1H-indole-1-carboxylic acid (F4)

[0234]

[0235] Di-tert-butyl dicarbonate (361 mg, 1.65 mmol) was added to 4 mL of an acetonitrile solution of indole-5-carboxaldehyde (200 mg, 1.38 mmol) with stirring, followed by the addition of 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 rapid column chromatography (ethyl acetate:hexane = 1:5) to obtain tert-butyl 3-formyl-1H-indole-1-carboxylate (Stage 1).

[0236] With stirring, thionyl chloride (2.0 mL, 28.05 mmol) and dimethylformamide (10 drops) were added sequentially to a 0.1 M dichloromethane solution of 4-tert-butyl benzoate (500 mg, 2.81 mmol). The mixture was heated for 12 hours and the solvent was evaporated under reduced pressure to obtain the intermediate chloride (stage 2). Glycine (180 mg, 2.398 mmol) and the intermediate chloride (0.542 mL, 2.758 mmol) were added sequentially to a 10% NaOH aqueous solution (23.98 mL) under reduced pressure at room temperature, and the mixture was stirred for 2 hours. The mixture was acidified to pH 2 with HCl and filtered through a funnel to obtain the resulting solid. The solid was washed with dichloromethane to provide 2-(4-(tert-butyl)benzoylamino)acetate (stage 3). 2-(4-(tert-butyl)benzoylamino)acetate (200 mg, 0.85 mmol) and 1-ethyl-3-(3-dimethylamino)propylcarbodiimide hydrochloride (179.25 mg, 0.935 mmol) were added sequentially to dichloromethane (8.5 mL) with stirring at room temperature. Then, tert-butyl 3-formyl-1H-indole-1-carboxylate (208 mg, 0.085 mmol) and triacetamide (0.208 mL, 2.550 mmol) were added sequentially to the mixture with stirring at room temperature, and the mixture was stirred for 12 hours. The solvent was removed under reduced pressure at room temperature, and the solid was obtained by filtration through a filter funnel. The solid was washed with methanol to provide the desired product (3-((2-(4-(tert-butyl)phenyl)-5-oxo) tert-butyl azole-4(5H)-methylene)-1H-indole-1-carboxylic acid, F4).

[0237] 1H NMR (400MHz, DMSO-d6) δ8.84(s,1H),8.31(d,J=7.6Hz,1H),8.09(d,J=7.9Hz,1H),7. 99(d,J=8.4Hz,2H),7.67-7.60(m,3H),7.45-7.33(m,2H),1.67(s,9H),1.31(s,9H).

[0238] Example 1.5: 4-(5-oxo-4-(thiophen-2-ylmethylene)-4,5-dihydro (F5)-2-azolylbenzylnitrile

[0239]

[0240] Glycine (180 mg, 2.40 mmol) and 4-cyanobenzoyl chloride (460 mg, 2.76 mmol) were added sequentially to 10% NaOH solution (24 mL) with stirring at room temperature, and the mixture was stirred at this temperature for 2 hours. The mixture was acidified with HCl until pH 2 was reached, and the resulting solid (2-(4-cyanobenzoylamino)acetate) was filtered through a filter funnel and then washed with dichloromethane. 2-(4-cyanobenzoylamino)acetate (400 mg, 1.96 mmol) and 1-ethyl-3-(3-dimethylamino)propylcarbodiimide hydrochloride (410 mg, 2.14 mmol) were added sequentially to dichloromethane (20 mL) with stirring at room temperature, and the mixture was stirred for 12 hours. Thiophene-2-carboxaldehyde (200 mg, 1.783 mmol) and triethylamine (0.5 mL, 3.57 mmol) were added sequentially to the solution under stirring at room temperature, and the mixture was stirred for 12 hours. The solvent was removed under reduced pressure, and the solid was obtained by filtration through a filter funnel. The solid was washed with methanol to provide the desired product (4-(5-oxo-4-(thiophene-2-ylmethylene)-4,5-dihydro) (Azol-2-yl)benzyl nitrile, F5).

[0241] 1 H NMR (400MHz, DMSO-d6) δ8.18 (d, 2H, J = 8.4Hz), 8.07 (t, 3H, J = 7.6Hz), 7.87 (d, 1H, J = 3.6Hz), 7.81 (s, 1H), 7.25 (t, 1H, J = 4.4Hz).

[0242] Example 1.6: 2-(4-(tert-butyl)phenyl)-4-((1-methyl-1H-pyrrolo-2-yl)methylene) Azolium-5(4H)-one (F6)

[0243]

[0244] 1 H NMR (400MHz, DMSO-d6) δ7.95 (d, 2H, J = 8.4Hz), 7.58 (d, 3H, J = 8.0Hz), 7.24 (s, 1H), 7.17 (s, 1H), 6.33 (t, 1H, J = 3.0Hz), 3.78 (s, 3H), 1.29 (s, 9H).

[0245] ESI(m / z) 309(MH) + )

[0246] Example 1.7: 4-(5-oxo-4-(thiophen-2-ylmethylene)-4,5-dihydro Methyl 2-(azolyl)benzoate (F7)

[0247]

[0248] 1 H NMR (400MHz, DMSO-d6) δ 8.16 (q, 4H, J = 8.8Hz), 8.08 (d, 1H, J = 5.2Hz), 8.02 (s, 1H), 7.86 (d, 1H, J = 3.6Hz), 7.79 (s, 1H), 3.88 (s, 3H).

[0249] ESI(m / z) 314(MH) + )

[0250] Example 1.8: 2-(4-(tert-butyl)phenyl)-4-((4-methylthiazolyl-5-yl)methylene) Azolium-5(4H)-one (F8)

[0251]

[0252] 1 H NMR (400MHz, DMSO-d6) δ9.28 (s, 1H), 7.95 (d, 2H, J = 8.4Hz), 7.62 (d, 2H, J = 8.4Hz), 7.53 (s, 1H), 2.59 (s, 3H), 1.29 (s, 9H).

[0253] ESI(m / z) 327(MH) + ).

[0254] Example 1.9: 2-(4-(tert-butyl)phenyl)-4-(thiophene-3-ylmethylene) Azolium-5(4H)-one (F9)

[0255]

[0256] 1 H NMR (400MHz, DMSO-d6) δ 8.44 (d, 1H, J = 2.4Hz), 8.01-7.99 (m, 3H), 7.71-7.69 (m, 1H), 7.59 (d, 2H, J = 8.8Hz), 7.37 (s, 1H), 1.29 (s, 9H).

[0257] ESI(m / z) 312(MH) + ).

[0258] Example 1.10: 4-((1H-pyrrolo-2-yl)methylene)-2-(4-(tert-butyl)phenyl) Azolium-5(4H)-one (F10)

[0259]

[0260] 1 H NMR (400MHz, DMSO-d6) δ11.53(s,1H),8.08(d,2H,J=8.2Hz),7.58(d,2H,J=8.3Hz),7.32(s,1H),7.19(s,1H),7.06(s,1H),6.34(s,1H),1.29(s,9H).

[0261] ESI(m / z) 295(MH) + ).

[0262] Example 1.11: 4-(benzo[b]thiophene-2-ylmethylene)-2-(4-(tert-butyl)phenyl) Azolium-5(4H)-one (F11)

[0263]

[0264] 1 H NMR (400MHz, DMSO-d6) δ9.23 (s, 1H), 8.31 (d, 1H, J = 8.0Hz), 8.09 (d, 3H, J = 8.4Hz), 7.64-7.62 (m, 3H), 7.53-7.44 (m, 2H), 1.31 (s, 9H).

[0265] ESI(m / z) 362(MH) + ).

[0266] Example 1.12: 4-((1H-pyrrolo-2-yl)methylene)-2-(4-isopropylphenyl) Azolium-5(4H)-one (F12)

[0267]

[0268] 1 H NMR (400MHz, DMSO-d6) δ11.52 (s, 1H), 8.08 (d, 2H, J = 8.0Hz), 7.43 (d, 2H, J = 8.0Hz), 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=6.8Hz).

[0269] ESI(m / z) 281(MH) + ).

[0270] Example 1.13: 2-(4-(tert-butyl)phenyl)-4-(quinoline-4-ylmethylene) Azolium-5(4H)-one (F13)

[0271]

[0272] 1 H NMR (400MHz, DMSO-d6) δ9.08 (d, 1H, J = 4.4Hz), 8.64 (d, 1H, J = 4.8Hz), 8.44 (d, 1H, J = 8.4Hz), 8.10-8.07 (m,3H),7.95(s,1H),7.83(t,1H,J=7.6Hz),7.71(d,1H,J=8.0Hz),7.66(d,2H,J=8.0Hz),1.31(s,9H).

[0273] ESI(m / z) 357(MH) + ).

[0274] Example 1.14: 2-(4-isobutylphenyl)-4-(thiophene-2-ylmethylene) Azolium-5(4H)-one (F14)

[0275]

[0276] 1H NMR (400MHz, DMSO-d6) δ8.01(d,1H,J=5.2Hz),7.96(d,2H,J=7.2Hz),7.81(d,1H,J=3.2Hz),7.67(s,1H),7. 39 (d, 2H, J = 7.2Hz), 7.22 (t, 1H, J = 4.4Hz), 2.53 (d, 2H, J = 6.8Hz), 1.91-1.81 (m, 1H), 0.84 (d, 6H, J = 6.4Hz).

[0277] ESI(m / z) 312(MH) + ).

[0278] Example 1.15: 2-(4-isobutylphenyl)-4-(thiophene-3-ylmethylene) Azolium-5(4H)-one (F15)

[0279]

[0280] 1 H NMR (400MHz, DMSO-d6) δ8.45 (d, 1H, J = 2.4Hz), 8.01-7.99 (m, 3H), 7.69 (q, 1H, J = 2.8Hz), 7 .39(s,1H),7.37(s,2H),2.53(d,2H,J=7.2Hz),1.92-1.81(m,1H),0.85(d,6H,J=6.8Hz).

[0281] ESI(m / z) 312(MH) + ).

[0282] Example 1.16: 4-(benzo[b]thiophene-3-ylmethylene)-2-(4-isopropylphenyl) Azolium-5(4H)-one (F16)

[0283]

[0284] 1 H NMR (400MHz, DMSO-d6) δ9.24 (s, 1H), 8.30 (d, 1H, J = 7.6Hz), 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 = 6.8Hz).

[0285] Example 1.17: 4-((1H-pyrrolo-2-yl)methylene)-2-(4-(trifluoromethyl)phenyl) Azolium-5(4H)-one (F17)

[0286]

[0287] 1 H NMR (400MHz, DMSO-d6) δ 11.67 (s, 1H), 8.39 (d, 2H, J = 7.6Hz), 7.95 (d, 2H, J = 8.0Hz), 7.42 (s, 1H), 7.33 (s, 1H), 7.15 (s, 1H), 6.41 (s, 1H).

[0288] ESI(m / z) 305(MH) - ),307(MH + )

[0289] Example 1.18: 4-(thiophene-3-ylmethylene)-2-(4-(trifluoromethyl)phenyl) Azolium-5(4H)-one (F18)

[0290]

[0291] 1 H NMR (400MHz, DMSO-d6) δ 8.55 (d, 1H, J = 1.9Hz), 8.32 (d, 2H, J = 8.1Hz), 8.07 (d, 1H, J = 5.0Hz), 7.98 (d, 2H, J = 8.2Hz), 7.78-7.74 (m, 1H), 7.53 (s, 1H).

[0292] Example 1.19: 4-(furan-2-ylmethylene)-2-(4-isobutylphenyl) Azolium-5(4H)-one (F19)

[0293]

[0294] 1 H NMR (400MHz, DMSO-d6) δ8.04(s,1H),7.97(d,2H,J=8.0Hz),7.55(d,1H,J=3.6Hz),7.36(d,2H,J=8.0H z),7.14(s,1H),6.79(d,1H,J=2.0Hz),2.52(d,2H,J=6.4Hz),1.90-1.80(m,1H),0.84(d,6H,J=6.4Hz)

[0295] Example 1.20: 4-((1H-pyrrolo-2-yl)methylene)-2-(4-isobutylphenyl) Azolium-5(4H)-one (F20)

[0296]

[0297] 1 H NMR (400MHz, DMSO-d6) δ11.51(s,1H),8.06(d,2H,J=8.0Hz),7.34(d,2H,J=8.4Hz),7.31(s,1H),7.1 8(s,1H),7.04(s,1H),6.33(s,1H),2.51(d,2H,J=6.8Hz),1.90-1.80(m,1H),0.84(d,6H,J=6.4Hz).

[0298] Example 1.21: 4-(benzo[b]thiophene-3-ylmethylene)-2-(4-isobutylphenyl) Azolium-5(4H)-one (F21)

[0299]

[0300] 1 H NMR (400MHz, DMSO-d6) δ9.24 (s, 1H), 8.29 (d, 1H, J = 8.0Hz), 8.08 (d, 3H, J = 8.0Hz), 7.60 (s, 1H), 7.52 -7.43(m,2H),7.39(d,2H,J=7.6Hz),2.54(d,2H,J=6.8Hz),1.89-1.84(m,1H),0.85(d,6H,J=6.4Hz).

[0301] Example 1.22: 2-(4-bromophenyl)-4-((4-methylthiazolyl)methylene) Azolium-5(4H)-one (F22)

[0302]

[0303] 1 H NMR (400MHz, DMSO-d6) δ9.31 (s, 1H), 7.96 (d, 2H, J = 8.4Hz), 7.83 (d, 2H, J = 8.4Hz), 7.63 (s, 1H), 2.62 (s, 3H).

[0304] ESI(m / z) 350(MH) + ).

[0305] Example 1.23: 2-Cyclohexyl-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F23)

[0306]

[0307] 1 H NMR (400MHz, DMSO-d6) δ7.96 (d, 1H, J = 5.2Hz), 7.74 (d, 1H, J = 3.6Hz), 7.57 (s, 1H), 7.18 (t, 1H, J = 4.2Hz), 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).

[0308] ESI(m / z) 262(MH) + ).

[0309] Example 1.24: 2-([1,1'-biphenyl]-4-yl)-4-((4-methylthiazolyl-5-yl)methylene) Azolium-5(4H)-one (F24)

[0310]

[0311] 1 H NMR (400MHz, DMSO-d6) δ9.31 (s, 1H), 8.12 (d, 2H, J = 8.4Hz), 7.93 (d, 2H, J = 8.0Hz), 7.77 ( d, 2H, J = 7.6Hz), 7.61 (s, 1H), 7.50 (t, 2H, J = 7.4Hz), 7.43 (t, 1H, J = 7.4Hz), 2.63 (s, 3H).

[0312] Example 1.25: 4-((1H-pyrrolo-2-yl)methylene)-2-([1,1'-biphenyl]-4-ylmethyl) Azolium-5(4H)-one (F25)

[0313]

[0314] 1 H NMR (400MHz, DMSO-d6) δ11.44 (s, 1H), 7.63 (d, 4H, J = 8.0Hz), 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).

[0315] ESI(m / z) 329(MH) + ),327(MH- ).

[0316] Example 1.26: 4-((1H-pyrrolo-2-yl)methylene)-2-(4-butylphenyl) Azolium-5(4H)-one (F26)

[0317]

[0318] 1 H NMR (400MHz, DMSO-d6) δ11.56(s,1H),8.11(d,2H,J=7.9Hz),7.43(d,2H,J=8.0Hz),7.35(s,1H),7.22(s,1H), 7.08(s,1H),6.37(s,1H),2.69(t,2H,J=7.6Hz),1.65-1.55(m,2H),1.38-1.27(m,2H),0.91(t,3H,J=7.3Hz).

[0319] Example 1.27: 2-(4-(tert-butyl)phenyl)-4-((1-phenyl-1H-pyrrolo-2-yl)methylene) Azolium-5(4H)-one (F27)

[0320]

[0321] 1 H NMR (400MHz, DMSO-d6) δ7.98 (d, 2H, J = 8.4Hz), 7.77 (d, 1H, J = 3.9Hz), 7.62-7.57 (m, 4H), 7.55-7.51 (m,1H),7.50-7.48(m,1H),7.45(d,2H,J=7.5Hz),6.74(s,1H),6.58(t,1H,J=3.4Hz),1.30(s,9H).

[0322] Example 1.28: 2-([1,1'-biphenyl]-4-ylmethyl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F28)

[0323]

[0324] 1H NMR (400MHz, DMSO-d6) δ7.98 (d, 1H, J = 4.8Hz), 7.76 (d, 1H, J = 3.6Hz), 7.65 (s, 2H), 7.6 3(s,3H),7.46-7.41(m,4H),7.33(t,1H,J=7.2Hz),7.19(t,1H,J=4.4Hz),4.10(s,2H).

[0325] Example 1.29: 2-(2,3-dihydro-1H-inden-5-yl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F29)

[0326]

[0327] 1 H NMR (400MHz, DMSO-d6) δ8.01(d,1H,J=5.2Hz),7.89(s,1H),7.83(d,1H,J=7.6Hz),7.80(d,1H,J=3.6 Hz), 7.65 (s, 1H), 7.43 (d, 1H, J = 8.0Hz), 7.23-7.21 (m, 1H), 2.93 (t, 4H, J = 7.3Hz), 2.08-2.00 (m, 2H).

[0328] ESI(m / z) 296(MH) + ).

[0329] Example 1.30: 4-((1H-pyrrolo-2-yl)methylene)-2-(naphth-1-yl) Azoxyl-5(4H)-one (F30)

[0330]

[0331] 1 H NMR (400MHz, DMSO-d6) δ11.62(s,1H),9.31(d,1H,J=8.0Hz),8.29(d,1H,J=7.2Hz),8.20(d,1H,J=8.4Hz),8.06 (d, 1H, J = 8.0Hz), 7.76 (t, 1H, J = 7.6Hz), 7.65 (q, 2H, J = 8.4Hz), 7.36 (s, 1H), 7.28 (s, 2H), 6.43 (t, 1H, J = 2.8Hz).

[0332] ESI(m / z) 289(MH) + ).

[0333] Example 1.31: 4-(4-(furan-2-ylmethylene)-5-oxo-4,5-dihydro Methyl 2-azolyl benzoate (F31)

[0334]

[0335] 1 H NMR(400MHz,DMSO-d6)δ8.21(s,1H),8.18(s,1H),8.13(s,1H),8.11-8.10( m,2H),7.60(d,1H,J=3.2Hz),7.27(s,1H),6.84-6.82(m,1H),3.87(s,3H).

[0336] Example 1.32: 4-(5-oxo-4-(thiophene-3-ylmethylene)-4,5-dihydro Methyl 2-azolyl benzoate (F32)

[0337]

[0338] 1 H NMR (400MHz, DMSO-d6) δ8.47(s,1H),8.16-8.00(m,5H),7.70(s,1H),7.44(s,1H),3.85(s,3H).

[0339] Example 1.33: N-(4-(5-oxo-4-(thiophene-2-ylmethylene)-4,5-dihydro) (F33)-Azol-2-yl)phenyl)acetamide

[0340]

[0341] Example 1.34: 2-(2-methoxyphenyl)-4-(thiophene-2-ylmethylene) Azoxyl-5(4H)-one (F34)

[0342]

[0343] Example 1.35: N-(tert-butyl)-4-(5-oxo-4-(thiophene-2-ylmethylene)-4,5-dihydro (F35)-2-yl)benzenesulfonamide

[0344]

[0345] Example 1.36: N-Isopropyl-4-(5-oxo-4-(thiophen-2-ylmethylene)-4,5-dihydro (F36)-2-yl)benzenesulfonamide

[0346]

[0347] Example 1.37: 2-([1,1'-biphenyl]-4-yl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F37)

[0348]

[0349] 1 H NMR (400MHz, DMSO-d6) δ8.23(d,2H,J=8.3Hz),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).

[0350] ESI(m / z) 332(MH) + ).

[0351] Example 1.38: 2-(4-(tert-butyl)phenyl)-4-(furan-2-ylmethylene) Azolium-5(4H)-one (F38)

[0352]

[0353] 1 H NMR (400MHz, DMSO-d6) δ 8.05 (s, 1H), 7.99 (d, 2H, J = 8.4Hz), 7.60 (d, 2H, J = 8.4Hz), 7.55 (d, 1H, J = 3.2Hz), 7.15 (s, 1H), 6.81 (br s, 1H), 1.29 (s, 9H).

[0354] ESI(m / z) 296(MH) + ).

[0355] Example 1.39: 2-(4-(tert-butyl)phenyl)-4-((1-methyl-1H-pyrazole-4-yl)methylene) Azolium-5(4H)-one (F39)

[0356]

[0357] 1H NMR (400MHz, DMSO-d6) δ 8.52 (s, 1H), 8.18 (s, 1H), 8.05 (d, 2H, J = 8.5Hz), 7.63 (d, 2H, J = 8.5Hz), 7.35 (s, 1H), 3.96 (s, 3H), 1.34 (s, 9H).

[0358] ESI(m / z) 310(MH) + ).

[0359] Example 1.40: 2-(4-(tert-butyl)phenyl)-4-(pyridin-3-ylmethylene) Azoxyl-5(4H)-one (F40)

[0360]

[0361] 1 H NMR (400MHz, DMSO-d6) δ9.28 (d, 1H, J = 1.9Hz), 8.78 (dt, 1H, J = 8.1, 1.7Hz), 8.65 (dd, 1H, J = 4 .8,1.6Hz),8.09(d,2H),7.68(d,2H,J=8.6Hz),7.61-7.56(m,1H),7.40(s,1H),1.34(s,9H).

[0362] ESI(m / z) 307(MH) + ).

[0363] Example 1.41: 2-(4-(tert-butyl)phenyl)-4-(pyridin-4-ylmethylene) Azolium-5(4H)-one (F41)

[0364]

[0365] 1 H NMR (400MHz, DMSO-d6) δ 8.71 (d, 2H, J = 5.2Hz), 8.13 (d, 2H, J = 5.5Hz), 8.07 (d, 2H), 7.66 (d, 2H, J = 8.3Hz), 7.27 (s, 1H), 1.31 (s, 9H).

[0366] ESI(m / z) 307(MH) + ).

[0367] Example 1.42: 2-([1,1'-biphenyl]-4-yl)-4-(pyridin-3-ylmethylene) Azolium-5(4H)-one (F42)

[0368]

[0369] 1 H NMR (400MHz, DMSO-d6) δ9.25 (s, 1H), 8.78 (d, 1H, J = 7.8Hz), 8.63 (s, 1H), 8.19 (d, 2H, J = 7.8Hz), 7.92 (d, 2H, J = 7.9Hz), 7.77 (d, 2H, J = 7.1Hz), 7.61-7.54 (m, 1H), 7.50 (t, 2H, J = 6.9Hz), 7.47-7.36 (m, 2H).

[0370] ESI(m / z) 327(MH) + ).

[0371] Example 1.43: 2-(4-(tert-butyl)phenyl)-4-(4-(methylthio)benzyl) Azolium-5(4H)-one (F43)

[0372]

[0373] 1 H NMR (400MHz, DMSO-d6) δ8.20(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).

[0374] ESI(m / z) 352(MH) + ).

[0375] Example 1.44: 4-Benzylene-2-(4-(tert-butyl)phenyl) Azoxyl-5(4H)-one (F44)

[0376]

[0377] 1 H NMR (400MHz, DMSO-d6) δ8.26 (d, 2H, J = 6.8Hz), 8.00 (d, 2H, J = 8.4Hz), 7.61 (d, 2H, J = 8.0Hz), 7.52-7.45 (m, 3H), 7.28 (s, 1H), 1.29 (s, 9H).

[0378] ESI(m / z) 306(MH) + ).

[0379] Example 1.45: 2-(4-bromophenyl)-4-(thiophene-2-ylmethylene) Azolium-5(4H)-one (F45)

[0380]

[0381] 1 H NMR (400MHz, DMSO-d6) δ8.05 (s, 1H), 7.95 (d, 2H, J = 7.6Hz), 7.83 (s, 2H), 7.81 (s, 1H), 7.73 (s, 1H), 7.23 (s, 1H).

[0382] Example 1.46: 2-(4-isopropylphenyl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F46)

[0383]

[0384] 1 H NMR (400MHz, DMSO-d6) δ8.02(d,1H,J=5.2Hz),7.97(d,2H,J=8.0Hz),7.81(d,1H,J=3.6Hz),7.6 7 (s, 1H), 7.48 (d, 2H, J = 8.4Hz), 7.22 (t, 1H, J = 4.4Hz), 3.02-2.92 (m, 1H), 1.21 (d, 6H, J = 6.8Hz).

[0385] ESI(m / z) 298(MH) + ).

[0386] Example 1.47: 2-(4-isopropylphenyl)-4-(thiophene-3-ylmethylene) Azolium-5(4H)-one (F47)

[0387]

[0388] 1 H NMR (400MHz, DMSO-d6) δ8.45 (d, 1H, J = 2.8Hz), 8.02-8.00 (m, 3H), 7.70 (q, 1H, J = 2. 8Hz), 7.47 (d, 2H, J = 7.2Hz), 7.38 (s, 1H), 3.03-2.93 (m, 1H), 1.21 (d, 6H, J = 6.8Hz).

[0389] ESI(m / z) 298(MH) + ).

[0390] Example 1.48: 4-(furan-2-ylmethylene)-2-(4-(trifluoromethyl)phenyl) Azolium-5(4H)-one (F48)

[0391]

[0392] 1 H NMR (400MHz, DMSO-d6) δ8.25 (d, 2H, J = 8.2Hz), 8.10 (s, 1H), 7.94 (d, 2H, J = 8.3Hz), 7.60 (d, 1H, J = 3.5Hz), 7.28 (s, 1H), 6.83-6.81 (m, 1H).

[0393] Example 1.49: 4-(furan-2-ylmethylene)-2-(4-isopropylphenyl) Azolium-5(4H)-one (F49)

[0394]

[0395] 1 H NMR (400MHz, DMSO-d6) δ8.04(s,1H),7.97(d,2H,J=8.1Hz),7.54(d,1H,J=3.3Hz),7.44(d ,2H,J=8.1Hz),7.13(s,1H),6.83-6.76(m,1H),3.01-2.88(m,1H),1.19(d,6H,J=6.9Hz).

[0396] Example 1.50: 2-(4-bromophenyl)-4-(furan-2-ylmethylene) Azoxyl-5(4H)-one (F50)

[0397]

[0398] 1 H NMR (400MHz, DMSO-d6) δ8.11(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).

[0399] Example 1.51: 2-(4-bromophenyl)-4-(thiophene-3-ylmethylene) Azolium-5(4H)-one (F51)

[0400]

[0401] 1H NMR (400MHz, DMSO-d6) δ 8.47 (d, 1H, J = 1.6Hz), 8.01-7.99 (m, 3H), 7.80 (s, 1H), 7.78 (s, 1H), 7.70-7.68 (m, 1H), 7.43 (s, 1H).

[0402] Example 1.52: 2-(naphthyl-2-yl)-4-(thiophene-2-ylmethylene) Azolium-5(4H)-one (F52)

[0403]

[0404] 1 H NMR (400MHz, DMSO-d6) δ8.70 (s, 1H), 8.52 (d, 1H, J = 2.4Hz), 8.21-8.17 (m, 2H), 8.12 (s, 1H), 8. 09(t,1H,J=4.8Hz),8.02(d,1H,J=8.0Hz),7.74-7.72(m,1H),7.69-7.61(m,2H),7.44(s,1H).

[0405] ESI(m / z) 306(MH) + ).

[0406] Example 1.53: 2-(4-Butylphenyl)-4-(thiophene-2-ylmethylene) Azolium-5(4H)-one (F53)

[0407]

[0408] 1 H NMR (400MHz, DMSO-d6) δ8.02(d,1H,J=5.0Hz),7.96(d,2H,J=8.2Hz),7.81(d,1H,J=3.6Hz),7.67(s,1H),7.42(d,2 H,J=8.2Hz),7.25-7.20(m,1H),2.66(t,2H,J=7.7Hz),1.61-1.51(m,2H),1.37-1.22(m,2H),0.87(t,3H,J=7.3Hz).

[0409] Example 1.54: 2-(4-Butylphenyl)-4-(furan-2-ylmethylene) Azolium-5(4H)-one (F54)

[0410]

[0411] 1 H NMR (400MHz, DMSO-d6) δ8.09 (d, 1H, J = 1.5Hz), 8.02 (d, 2H, J = 8.2Hz), 7.60 (d, 1H, J = 3.5Hz), 7.45 (d, 2H, J = 8.2Hz), 7.19(s,1H),6.86-6.83(m,1H),2.69(t,2H,J=7.7Hz),1.66-1.54(m,2H),1.39-1.27(m,2H),0.91(t,3H,J=7.3Hz).

[0412] Example 1.55: 2-(4-(difluoromethoxy)phenyl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F55)

[0413]

[0414] 1 H NMR (400MHz, DMSO-d6) δ8.10 (d, 2H, J = 8.8Hz), 8.03 (d, 1H, J = 5.2Hz), 7.82 (d, 1H ,J=3.6Hz),7.69(s,1H),7.41(s,1H),7.38(d,2H,J=8.8Hz),7.24-7.22(m,1H).

[0415] Example 1.56: 2-(4-(tert-butyl)phenyl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F56)

[0416]

[0417] 1 H NMR (400MHz, DMSO-d6) δ8.02 (d, 1H, J = 5.2Hz), 7.96 (d, 2H, J = 7.6Hz), 7.80 (d, 1H, J=3.6Hz),7.66(s,1H),7.61(d,2H,J=7.6Hz),7.22(t,1H,J=4.0Hz),1.30(s,9H).

[0418] 13 C NMR(400MHz,DMSO-d6)δ166.7,162.1,157.2,137.6,136.9,136.6,130.7, 129.6,129.1,129.1,126.4,126.4,124.9,122.8,35.46,31.2,31.2,31.2.

[0419] Example 1.57: 4-(2-nitrobenzylidene)-2-phenyl Azolium-5(4H)-one (F57)

[0420]

[0421] Example 1.58: 4-((2-(4-(tert-butyl)phenyl)-5-oxo Azole-4(5H)-methylene)phenylacetate (F58)

[0422]

[0423] Example 1.59: 2-(3-chlorobenzo[b]thiophen-2-yl)-4-(4-(dimethylamino)benzylidene) Azolium-5(4H)-one (F59)

[0424]

[0425] Example 1.60: 4-((2-(4-methoxyphenyl)-5-oxo Azole-4(5H)-methylene)benzoate (F60)

[0426]

[0427] Example 1.61: 2-Phenylacetyl-4-(thiophene-2-ylmethylene) Azolium-5(4H)-one (F61)

[0428]

[0429] Example 1.62: 4-(4-isopropylbenzyl)-2-(naphth-1-yl) Azolium-5(4H)-one (F62)

[0430]

[0431] Example 1.63: 2-(3-iodo-4-methylphenyl)-4-(thiophene-2-ylmethylene) Azolium-5(4H)-one (F63)

[0432]

[0433] Example 1.64: 4-(4-(benzyloxy)benzylene)-2-(4-(tert-butyl)phenyl) Azolium-5(4H)-one (F64)

[0434]

[0435] Example 1.65: 4-((1-(2-chlorobenzyl)-1H-pyrrolo-2-yl)methylene)-2-phenyl Azolium-5(4H)-one (F65)

[0436]

[0437] Example 1.66: N-(4-(4-(2-(allyloxy)-4-(diethylamino)benzylidene)-5-oxo-4,5-dihydro (F66)-2-(azolyl)phenyl)-5-bromonicotinamide

[0438]

[0439] Example 1.67: 4-((2-(2-chlorophenyl)-5-oxo Azole-4(5H)-ylidene)methyl)phenyl-4-(tert-butyl)benzoate (F67)

[0440]

[0441] Example 1.68: 4-((Z)-1-(3-ethyl-5-methoxybenzo[d]thiazol-2(3H)-ylidene)but-2-ylidene)-2-phenyl Azolium-5(4H)-one (F68)

[0442]

[0443] Example 1.69: 4-((2-(4-acetaminophenyl)-5-oxo Azole-4(5H)-methylene)-2-ethoxyphenylthiophene-2-carboxylate (F69)

[0444]

[0445] Example 1.70: 4-((6-ethoxy-2-(phenylthio)quinoline-3-yl)methylene)-2-phenyl Azoxyl-5(4H)-one (F70)

[0446]

[0447] Example 1.71: 2-(3-bromophenyl)-4-((5-chloro-3-methyl-1-phenyl-1H-pyrazol-4-yl)methylene) Azolium-5(4H)-one (F71)

[0448]

[0449] Example 1.72: 4-((1-acetyl-1H-indol-3-yl)methylene)-2-(4-bromophenyl) Azolium-5(4H)-one (F72)

[0450]

[0451] Example 1.73: 2-(4-(tert-butyl)phenyl)-4-((5-(3-(trifluoromethyl)phenyl)furan-2-yl)methylene) Azolium-5(4H)-one (F73)

[0452]

[0453] Example 1.74: 4-((7-methoxy-2-oxo-1,2-dihydroquinolin-3-yl)methylene)-2-phenyl Azoxyl-5(4H)-one (F74)

[0454]

[0455] Example 1.75: 2-(4-methoxyphenyl)-4-(2-(thiophene-2-yl)-4H-chromene-4-ylidene) Azolium-5(4H)-one (F75)

[0456]

[0457] Example 1.76: N,N-Dimethyl-3-((5-oxo-2-(p-tolyl)) azole-4(5H)-methylene)-1H-indole-1-sulfonamide (F76)

[0458]

[0459] Example 1.77: 2-((2-(4-chlorophenyl)-5-oxo Azole-4(5H)-methylene)phenyl-4-acetaminobenzenesulfonate (F77)

[0460]

[0461] Example 1.78: 4-((5-oxo-2-phenyl) Azole-4(5H)-methylene)phenylfuran-2-carboxylate (F78)

[0462]

[0463] Example 1.79: 2,2'-(1,4-phenylene)bis(4-((5-methylfuran-2-yl)methylene) Azolium-5(4H)-one (F79)

[0464]

[0465] Example 1.80: 2-((2-(4-(tert-butyl)phenyl)-5-oxo Azole-4(5H)-methylene)phenylbenzenesulfonate (F80)

[0466]

[0467] Example 1.81: 4-((1-acetyl-1H-indol-3-yl)methylene)-2-(4-(tert-butyl)phenyl) Azoxyl-5(4H)-one (F81)

[0468]

[0469] Example 1.82: 4-((1-acetyl-3-phenyl-1H-pyrazole-4-yl)methylene)-2-(4-(tert-butyl)phenyl) Azolium-5(4H)-one (F82)

[0470]

[0471] Example 1.83: 4-((1,3-diphenyl-1H-pyrazole-4-yl)methylene)-2-(p-tolyl) Azolium-5(4H)-one (F83)

[0472]

[0473] Example 1.84: 4-((6-methoxy-2-oxo-1,2-dihydroquinolin-3-yl)methylene)-2-phenyl Azolium-5(4H)-one (F84)

[0474]

[0475] Example 1.85: 4-(2,6-diphenyl-4H-thiaran-4-ylidene)-2-phenyl Azolium-5(4H)-one (F85)

[0476]

[0477] Example 1.86: N-(4-(4-(4-((2-chloroethyl)(methyl)amino)benzylidene)-5-oxo-4,5-dihydro (F86)-2-yl)phenyl)acetamide

[0478]

[0479] Example 1.87: 2-(4-methoxyphenyl)-4-(thiophene-2-ylmethylene) Azolium-5(4H)-one (F87)

[0480]

[0481] Example 1.88: 2-(4-chlorophenyl)-4-(thiophene-2-ylmethylene) Azolium-5(4H)-one (F88)

[0482]

[0483] Example 1.89: 2-(thien-2-yl)-4-(thien-2-ylmethylene) Azolium-5(4H)-one (F89)

[0484]

[0485] Example 1.90: 4-(5-oxo-4-(thiophen-2-ylmethylene)-4,5-dihydro (F90)-2-azolyl)phenylacetate

[0486]

[0487] Example 1.91: 2-(4-chloro-3-nitrophenyl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F91)

[0488]

[0489] Example 1.92: 2-(2-chlorophenyl)-4-(thiophene-2-ylmethylene) Azolium-5(4H)-one (F92)

[0490]

[0491] Example 1.93: 2-(2-chloro-4-nitrophenyl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F93)

[0492]

[0493] Example 1.94: 4-(thiophene-2-ylmethylene)-2-(3,4,5-trimethoxyphenyl) Azolium-5(4H)-one (F94)

[0494]

[0495] Example 1.95: 2-(4-phenoxyphenyl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F95)

[0496]

[0497] Example 1.96: 2-(4-(tert-butyl)phenyl)-4-((5-methylthiophen-2-yl)methylene) Azolium-5(4H)-one (F96)

[0498]

[0499] Example 1.97: 2-(benzo[d][1,3]dioxo-5-yl)-4-(thiophene-2-ylmethylene) Azolium-5(4H)-one (F97)

[0500]

[0501] Example 1.98: 2-(4-(2-oxopyrrolidone-1-yl)phenyl)-4-(thiophene-2-ylmethylene) Azolium-5(4H)-one (F98)

[0502]

[0503] Example 1.99: 4-((5-chlorothiophen-2-yl)methylene)-2-(4-methoxyphenyl) Azolium-5(4H)-one (F99)

[0504]

[0505] Example 1.100: 2-(benzo[d][1,3]dioxo-5-yl)-4-((5-(piperidin-1-yl)thiophen-2-yl)methylene) Azolium-5(4H)-one (F100)

[0506]

[0507] Example 1.101: 2-(3,4-diethoxyphenyl)-4-(thiophene-2-ylmethylene) Azolium-5(4H)-one (F101)

[0508]

[0509] Example 1.102: 2-(4-(difluoromethoxy)-3-methoxyphenyl)-4-(thiophene-2-ylmethylene) Azolium-5(4H)-one (F102)

[0510]

[0511] Example 1.103: 2-(5-ethyl-4-methylthiophen-2-yl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F103)

[0512]

[0513] Example 1.104: 2-(3-methoxyphenyl)-4-(thiophene-2-ylmethylene) Azolium-5(4H)-one (F104)

[0514]

[0515] Example 1.105: 4-((5-(dimethylamino)thiophen-2-yl)methylene)-2-(naphthyl-2-yl) Azolium-5(4H)-one (F105)

[0516]

[0517] Example 1.106: N-(4-(4-((4-bromothiophene-2-yl)methylene)-5-oxo-4,5-dihydro (F106)-Azol-2-yl)phenyl)acetamide

[0518]

[0519] Example 1.107: N-(4-(4-((5-bromothiophene-2-yl)methylene)-5-oxo-4,5-dihydro (F107)-Azol-2-yl)phenyl)acetamide

[0520]

[0521] Example 1.108: N-(4-(4-((5-(dimethylamino)thiophen-2-yl)methylene)-5-oxo-4,5-dihydro (F108)-2-yl)phenyl)acetamide

[0522]

[0523] Example 1.109: 2-(4-(benzyloxy)phenyl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F109)

[0524]

[0525] Example 1.110: 2-((E)-styryl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F110)

[0526]

[0527] Example 1.111: 2-(5-methylthiophen-2-yl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F111)

[0528]

[0529] Example 1.112: N,N-Diethyl-3-(5-oxo-4-(thiophen-2-ylmethylene)-4,5-dihydro (F112)-Azol-2-yl)benzenesulfonamide

[0530]

[0531] Example 1.113: 2-(2-methoxyphenyl)-4-((2,4,5-trimethylthiopheno[2,3-d]pyrimidin-6-yl)methylene) Azolium-5(4H)-one (F113)

[0532]

[0533] Example 1.114: 4-((5-(dimethylamino)thiophen-2-yl)methylene)-2-(2-methoxyphenyl) Azolium-5(4H)-one (F114)

[0534]

[0535] Example 1.115: 4-((5-methylthiophen-2-yl)methylene)-2-(thiophen-2-yl) Azolium-5(4H)-one (F115)

[0536]

[0537] Example 1.116: N,N-Dimethyl-3-(5-oxo-4-(thiophen-2-ylmethylene)-4,5-dihydro (F116)-2-ylbenzenesulfonamide

[0538]

[0539] Example 1.117: 4-((5-bromothiophene-2-yl)methylene)-2-(2-methoxyphenyl) Azolium-5(4H)-one (F117)

[0540]

[0541] Example 1.118: 4-((5-bromothiophen-2-yl)methylene)-2-(thiophen-2-yl) Azolium-5(4H)-one (F118)

[0542]

[0543] Example 1.119: 2-(5-methylthiophen-2-yl)-4-((5-methylthiophen-2-yl)methylene) Azolium-5(4H)-one (F119)

[0544]

[0545] Example 1.120: 4-((5-(piperidin-1-yl)thiophen-2-yl)methylene)-2-(thiophen-2-yl) Azolium-5(4H)-one (F120)

[0546]

[0547] Example 1.121: 4-((5-bromothiophene-2-yl)methylene)-2-(2-(difluoromethoxy)phenyl) Azolium-5(4H)-one (F121)

[0548]

[0549] Example 1.122: 2-(4-(tert-butyl)phenyl)-4-(2-(difluoromethoxy)benzyl) Azolium-5(4H)-one (F122)

[0550]

[0551] Example 1.123: 4-((1-(2-chlorobenzyl)-1H-pyrazol-4-yl)methylene)-2-(naphthyl-2-yl) Azolium-5(4H)-one (F123)

[0552]

[0553] Example 1.124: 2-(4-(tert-butyl)phenyl)-4-((1-phenyl-1H-pyrazole-4-yl)methylene) Azolium-5(4H)-one (F124)

[0554]

[0555] Example 1.125: 2-(2-methoxyphenyl)-4-((1-methyl-1H-pyrazol-4-yl)methylene) Azolium-5(4H)-one (F125)

[0556]

[0557] Example 1.126: N-(tert-butyl)-4-(4-((5-methylfuran-2-yl)methylene)-5-oxo-4,5-dihydro (F126)-2-yl)benzenesulfonamide

[0558]

[0559] Example 1.127: N,N-Diethyl-4-(4-(1-methylpyridin-2(1H)-ylidene)-5-oxo-4,5-dihydro (F127)-2-yl)benzenesulfonamide

[0560]

[0561] Example 1.128: 2-(4-((2-(2-methoxyphenyl)-5-oxo azole-4(5H)-methylene)phenoxy)acetamide (F128)

[0562]

[0563] Example 1.129: N-(4-((2-(4-isopropoxyphenyl)-5-oxo azole-4(5H)-methylene)phenyl)acetamide (F129)

[0564]

[0565] Example 1.130: 4-((1-benzyl-1H-pyrazole-4-yl)methylene)-2-(4-isopropoxyphenyl) Azolium-5(4H)-one (F130)

[0566]

[0567] Example 1.131: N,N-Diethyl-3-(4-((1-methyl-1H-pyrazol-4-yl)methylene)-5-oxo-4,5-dihydro (F131)-2-yl)benzenesulfonamide

[0568]

[0569] Example 1.132: 4-(4-(1H-1,2,4-thiazolyl-1-yl)benzylidene)-2-(2-iodophenyl) Azolium-5(4H)-one (F132)

[0570]

[0571] Example 1.133: 4-(4-((3,5-dimethylisocyanate) (Azol-4-yl)methoxy)-3-methoxybenzyl)-2-(2-methoxyphenyl) Azolium-5(4H)-one (F133)

[0572]

[0573] Example 1.134: N-(4-(4-(4-(1H-1,2,4-thiazo-1-yl)benzylidene)-5-oxo-4,5-dihydro (F134)-Azol-2-yl)phenyl)acetamide

[0574]

[0575] Example 1.135: 2-(2-(difluoromethoxy)phenyl)-4-((E)-3-(furan-2-yl)allylidel) Azolium-5(4H)-one (F135)

[0576]

[0577] Example 1.136: 4-((1-(tert-butyl)-1H-pyrazole-4-yl)methylene)-2-(2-methoxyphenyl) Azolium-5(4H)-one (F136)

[0578]

[0579] Example 1.137: 4-((4-methylthiazolyl-5-yl)methylene)-2-(4-propylphenyl) Azolium-5(4H)-one (F137)

[0580]

[0581] 1 H NMR (400MHz, DMSO-d6) δ9.27 (s, 1H), 9.94 (d, 2H, J = 8.0Hz), 7.53 (s, 1H), 7.41 (d, 2H, J =8.0Hz), 2.63 (t, 2H, J = 7.6Hz), 2.59 (s, 3H), 1.64-1.55 (m, 2H), 0.87 (t, 3H, J = 7.2Hz).

[0582] 13 C NMR (400MHz, DMSO-d6) δ166.1,12.6,160.62,159.76,149.2,132.0,129.9,129.3,126.5,123.0,121.3,37.7,24.1,16.2,14.0.

[0583] Example 1.138: 4-((1H-pyrrolo-2-yl)methylene)-2-(4-butoxyphenyl) Azolium-5(4H)-one (F138)

[0584]

[0585] 1 H NMR (400MHz, DMSO-d6) δ11.50 (s, 1H), 8.08 (d, 2H, J = 8.4Hz), 7.29 (s, 1H), 7.13 (s, 1H), 7.07 (d, 2H, J = 8.4Hz), 7.00 (s, 1H), 6.31 (s, 1H), 4.03 (t, 2H, J = 6.0Hz), 1.70-1.67 (m, 2H), 1.45-1.36 (m, 2H), 0.91 (t, 3H, J = 5.2Hz).

[0586] Example 1.139: 2-(4-hexylphenyl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F139)

[0587]

[0588] 1H NMR (400MHz, DMSO-d6) δ8.01(d,1H,J=3.6Hz),7.95(d,2H,J=8.0Hz),7.80(brs,1H),7.66(s,1H),7.41(d,2H,J=8.0Hz),7.22(br s, 1H), 2.64 (t, 2H, J = 7.2Hz), 1.56 (br s 2H), 1.24 (s, 6H), 0.81 (br s, 3H).

[0589] 13 C NMR (400MHz, DMSO-d6) δ166.7,162.2,149.2,137.6,136.9,136.6,130.7,129.7,128.6,128.2,124.8,123.0,35.7,31.5,30.9,28.7,22.5,14.4.

[0590] Example 1.140: 4-((1H-indol-2-yl)methylene)-2-(4-(tert-butyl)phenyl) Azolium-5(4H)-one (F140)

[0591]

[0592] 1 H NMR (400MHz, DMSO-d6) δ11.27 (s, 1H), 8.18 (d, 2H, J = 8.0Hz), 7.63 (t, 4H, J = 7.4Hz) ,7.35(d,2H,J=8.8Hz),7.25(t,1H,J=7.4Hz),7.05(t,1H,J=7.6Hz),1.30(s,9H).

[0593] 13 C NMR(400MHz,DMSO-d6)δ166.9,161.9,157.1,139.7,133.2,130.8,128.5,1 28.2,126.5,125.6,122.9,122.1,121.0,120.8,113.4,113.0,35.3,31.2.

[0594] Example 1.141: 4-(furan-2-ylmethylene)-2-(4-pentylphenyl) Azolium-5(4H)-one (F141)

[0595]

[0596] 1H NMR (400MHz, DMSO-d6) δ8.05 (s, 1H), 7.97 (d, 2H, J = 7.6Hz), 7.56-7.55 (m, 1H), 7.40 (d, 2H, J = 7.6Hz), 7.15 (s, 1H), 6.80 (br s,1H),2.64(t,2H,J=7.4Hz),1.59-1.57(m,2H),1.26(br s,4H),0.82(t,3H,J=6.2Hz).

[0597] 13 C NMR (400MHz, DMSO-d6) δ167.0,162.8,150.4,149.3,148.3,130.5,129.7,128.4,123.0,120.8,117.4,114.6,35.6,31.3,30.7,22.3,14.3.

[0598] Example 1.142: 2-(4-Butoxyphenyl)-4-(Thiophen-2-ylmethylene) Azolium-5(4H)-one (F142)

[0599]

[0600] *749 1 H NMR (400MHz, DMSO-d6) δ7.98-7.96 (m, 3H), 7.78 (d, 1H, J = 3.2Hz), 7.60 (s, 1H), 7.21 (t, 1H, J = 4.4Hz), 7. 12 (d, 2H, J = 8.4Hz), 4.05 (t, 2H, J = 6.4Hz), 1.73-1.66 (m, 2H), 1.46-1.37 (m, 2H), 0.91 (t, 3H, J = 7.2Hz).

[0601] 13 C NMR (400MHz, DMSO-d6) δ166.8,163.4,162.0,137.7,136.5,136.1,130.9,130.3,129.5,123.8,117.4,115.8,69.2,31.0,19.1,14.1.

[0602] Example 1.143: 4-((1H-pyrrolo-2-yl)methylene)-2-(4-propylphenyl) Azolium-5(4H)-one (F143)

[0603]

[0604] 1 H NMR (400MHz, DMSO-d6) δ11.52(s,1H),8.07(d,2H,J=8.0Hz),7.39(d,2H,J=8.4Hz),7.31(s,1H),7.19( s,1H),7.04(s,1H),6.34-6.32(m,1H),2.63(t,2H,J=8.4Hz),1.65-1.56(m,2H),0.88(t,3H,J=7.4Hz).

[0605] ESI(m / z) 281(MH) + ).

[0606] Example 1.144: 2-(4-propylphenyl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F144)

[0607]

[0608] 1 H NMR (400MHz, DMSO-d6) δ8.02 (d, 1H, J = 5.2Hz), 7.96 (d, 2H, J = 8.4Hz), 7.81 (d, 1H, J = 3.6Hz), 7.67 (s, 1H), 7.42 (d, 2H, J = 8.0Hz), 7.23-7.21 (m, 1H), 2.64 (t, 2H, J = 7.4Hz), 1.65-1.56 (m, 2H), 0.88 (t, 3H, J = 7.4Hz).

[0609] ESI(m / z) 298(MH) + ).

[0610] Example 1.145: 2-(4-propylphenyl)-4-(thiophen-3-ylmethylene) Azolium-5(4H)-one (F145)

[0611]

[0612] 1 H NMR (400MHz, DMSO-d6) δ8.02(d,1H,J=5.2Hz),7.97(d,2H,J=4.4Hz),7.81(d,1H,J=3.6Hz),7.67(s,1H), 7.42 (d, 2H, J = 8.4Hz), 7.23-7.21 (m, 1H), 2.63 (t, 2H, J = 8.4Hz), 1.65-1.56 (m, 2H), 0.88 (t, 3H, J = 7.2Hz).

[0613] ESI(m / z) 298(MH) + ).

[0614] Example 1.146: 4-(furan-2-ylmethylene)-2-(4-propylphenyl) Azolium-5(4H)-one (F146)

[0615]

[0616] 1 H NMR (400MHz, DMSO-d6) δ8.05(s,1H),7.98(d,2H,J=8.0Hz),7.56(s,1H),7.41(d,2H,J=8.0Hz ), 7.15 (s, 1H), 6.80 (s, 1H), 2.63 (t, 2H, J = 7.4Hz), 1.63-1.57 (m, 2H), 0.87 (t, 3H, J = 7.2Hz).

[0617] 13 C HMR (400MHz, DMSO-d6) δ167.0,162.8,150.4,149.0,148.3,130.5,129.8(2),128.4(2),123.0,120.8,117.4,114.6,37.7,24.1,14.0.

[0618] ESI(m / z) 282(MH) + ).

[0619] Example 1.147: 4-((1H-pyrrolo-2-yl)methylene)-2-(4-hexylphenyl) Azolium-5(4H)-one (F147)

[0620]

[0621] ESI(m / z) 323(MH) + ).

[0622] Example 1.148: 2-(4-(4-((1H-pyrrolo-2-yl)methylene)-5-oxo-4,5-dihydro (F148)-2-azolyl)phenyl)-2-methylacrylonitrile

[0623]

[0624] ESI(m / z) 306(MH) + ).

[0625] Example 1.149: 4-((1H-pyrrolo-2-yl)methylene)-2-(3,5-dimethylphenyl) Azolium-5(4H)-one (F149)

[0626]

[0627] ESI(m / z) 267(MH) + ).

[0628] Example 1.150: 2-(3,5-dimethylphenyl)-4-(thiophen-2-ylmethylene) Azolium-5(4H)-one (F150)

[0629]

[0630] 1 H NMR (400MHz, DMSO-d6) δ8.01(d,1H,J=4.8Hz),7.79(d,1H,J=3.2Hz),7.65(s,1H),7.63(s,1H),7.28(s,1H),7.21(t,1H,J=4.2Hz),2.33(s,6H)

[0631] ESI(m / z) 284(MH) + ).

[0632] Example 2.1: Methyl 2-(4-(tert-butyl)benzoamide)-3-(1H-pyrrolo-2-yl)acrylate (G1)

[0633]

[0634] 1 H NMR (400MHz, DMSO-d6) δ11.56(s,1H),9.73(s,1H),7.94(d,2H,J=8.1Hz),7.50(d,2H,J=8. 1Hz),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).

[0635] Example 2.2: Methyl 2-(4-(tert-butyl)benzoamide)-3-(thiophen-2-yl)acrylate (G2)

[0636]

[0637] 1H NMR (400MHz, DMSO-d6) δ9.77 (s, 1H), 7.95 (d, 2H, J = 8.3Hz), 7.88 (s, 1H), 7.70 (d ,1H,J=5.0Hz),7.57-7.50(m,3H),7.12-7.09(m,1H),3.69(s,3H),1.29(s,9H).

[0638] Example 2.3: 2-(4-(tert-butyl)benzamido)-3-(thiophen-2-yl)acrylate (G3)

[0639]

[0640] 1 H NMR (400MHz, DMSO-d6) δ12.62(s,1H),9.64(s,1H),7.93(d,2H,J=8.3Hz),7.83(s, 1H), 7.66 (d, 1H, J = 5.0Hz), 7.54-7.49 (m, 3H), 7.09 (t, 1H, J = 4.0Hz), 1.29 (s, 9H).

[0641] ESI(m / z)352(MNa + ),328(MH - ).

[0642] Example 2.4: Methyl 2-(4-(tert-butyl)benzamido)-3-(furan-2-yl)acrylate (G4)

[0643]

[0644] 2-(4-(tert-butyl)phenyl)-4-(furan-2-ylmethylene) Azoxyl-5(4H)-one (F38) (300 mg, 1.02 mmol) was added to MeOH (10 mL) with stirring. When the suspension became a clear solution, triethylamine (0.425 mL, 3.05 mmol) was added to the mixture, and the mixture was stirred for 1 hour. After the reaction was complete (TLC), the solvent was removed under reduced pressure. The solution was filtered through a filter funnel and recrystallized from methanol to provide the resulting solid (2-(4-(tert-butyl)benzamido)-3-(furan-2-yl)acrylate (G4).

[0645] 1H NMR (400MHz, DMSO-d6) δ9.87 (s, 1H), 7.92 (d, 2H, J = 8.3Hz), 7.81 (s, 1H), 7.51 (d, 2H, J = 8 .3Hz), 7.24 (s, 1H), 6.84 (d, 1H, J = 3.4Hz), 6.60-6.57 (m, 1H), 3.68 (s, 3H), 1.29 (s, 9H).

[0646] ESI(m / z) 328(MH) + ),350(MNa + ),326(MH - ).

[0647] Example 2.5: 2-(4-(tert-butyl)benzamido)-3-(furan-2-yl)acrylate (G5)

[0648]

[0649] 1 H NMR (400MHz, DMSO-d6) δ12.67(s,1H),9.72(s,1H),7.90(d,2H,J=8.2Hz),7.78(s,1H),7 .50(d,2H,J=8.3Hz),7.23(s,1H),6.78(d,1H,J=3.3Hz),6.58-6.55(m,1H),1.28(s,9H).

[0650] ESI(m / z)336(MNa + ),312(MH - ).

[0651] Example 2.6: 2-(4-(tert-butyl)benzamido)-3-(1H-pyrrolo-2-yl)tert-butyl acrylate (G6)

[0652]

[0653] 1 H NMR (400MHz, DMSO-d6) δ11.24(s,1H),9.49(s,1H),7.90(d,2H,J=8.3Hz),7.50(d,2H,J =8.3Hz),7.33(s,1H),6.94(s,1H),6.48(s,1H),6.10(s,1H),1.39(s,9H),1.29(s,9H).

[0654] Example 2.7: 2-(4-(tert-butyl)benzamido)-3-(1H-pyrrolo-2-yl)acrylate (G7)

[0655]

[0656] 4-((1H-pyrrolo-2-yl)methylene)-2-(4-(tert-butyl)phenyl) Azoxyl-5(4H)-one (F10) (1.02 mmol) was added to acetone (5 ml) with 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 complete (TLS), the mixture was acidified with 10% HCl and filtered through a filter funnel to give solid (2-(4-(tert-butyl)benzamido)-3-(1H-pyrrolo-2-yl)acrylate (G7).

[0657] 1 H NMR (400MHz, DMSO-d6) δ12.23 (s, 1H), 11.24 (s, 1H), 9.48 (s, 1H), 7.92 (d, 2H, J = 8.4Hz), 7.50(d,2H,J=8.0Hz),7.43(s,1H),6.95(s,1H),6.47(s,1H),6.11(s,1H),1.29(s,9H).

[0658] Example 2.8: 2-(4-(tert-butyl)benzamido)-3-(thiophen-3-yl)acrylate (G8)

[0659]

[0660] 1 H NMR (400MHz, DMSO-d6) δ 12.59 (S, 1H), 9.76 (s, 1H), 7.91 (d, 3H, J = 9.6Hz), 7.51 (d, 4H, J = 7.2Hz), 7.37 (d, 1H, J = 4.8Hz), 1.29 (s, 9H).

[0661] Example 2.9: 2-(4-(tert-butyl)benzamido)-3-(4-methylthiazolyl-5-yl)acrylate (G9)

[0662]

[0663] 1 H NMR (400MHz, DMSO-d6) δ12.83(s,1H),9.73(s,1H),9.00(s,1H),7.92(d,2H,J=8.4Hz),7.74(s,1H),7.53(d,2H,J=8.4Hz),2.47(s,3H),1.29(s,9H).

[0664] Example 2.10: 2-(4-(tert-butyl)benzamido)-3-(1-methyl-1H-pyrazole-4-yl)acrylate (G10)

[0665]

[0666] 1 H NMR (400MHz, DMSO-d6) δ12.39(s,1H),9.57(s,1H),7.95-7.88(m,3H),7.61(s,1H),7.51(d,2H,J=8.0Hz),7.39(s,1H),3.79(s,3H),1.29(s,9H).

[0667] ESI(m / z) 328(MH) + ),350(MNa + ),327(MH - ).

[0668] Example 2.11: Methyl 2-(4-Butylbenzamido)-3-(1H-pyrrolo-2-yl)acrylate (G11)

[0669]

[0670] 1 H NMR (400MHz, DMSO-d6) δ11.35(s,1H),9.64(s,1H),7.95(d,2H,J=6.7Hz),7.49(s,1H),7.34(d,2H,J=6.8Hz),7.02(s,1 H),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).

[0671] ESI(m / z) 327(MH) + ),349(MNa + ),325(MH - ).

[0672] Example 2.12: 2-(4-Butylbenzamido)-3-(thiophen-2-yl)acrylate (G12)

[0673]

[0674] 1H NMR (400MHz, DMSO-d6) δ12.67(s,1H),9.67(s,1H),7.95(d,2H,J=7.9Hz),7.87(s,1H),7.70(d,1H,J=4.9Hz),7.59-7.49(m,1H), 7.36 (d, 2H, J = 7.9Hz), 7.13 (d, 1H, J = 4.4Hz), 2.67 (t, 2H, J = 7.6Hz), 1.66-1.55 (m, 2H), 1.40-1.26 (m, 2H), 0.91 (t, 3H, J = 7.3Hz).

[0675] ESI(m / z) 330(MH) + ),328(MH - ).

[0676] Example 2.13: 2-(4-(tert-butyl)benzamido)-3-(1-phenyl-1H-pyrrolo-2-yl)acrylate (G13)

[0677]

[0678] 1 H NMR (400MHz, DMSO-d6) δ12.39 (s, 1H), 9.70 (s, 1H), 7.98 (d, 2H, J = 7.8Hz), 7.65-7.49 (m, 5H ),7.38(d,2H,J=7.4Hz),7.23(s,1H),7.16(s,1H),6.78(s,1H),6.34(s,1H),1.33(s,9H).

[0679] ESI (m / z) 389 (MH) + ),387(MH - ).

[0680] Example 2.14: 2-(4-(tert-butyl)benzamido)-3-(1-phenyl-1H-pyrrolo-2-yl)methyl acrylate (G14)

[0681]

[0682] 1 H NMR (400MHz, DMSO-d6) δ9.81 (s, 1H), 7.96 (d, 2H, J = 7.7Hz), 7.62-7.45 (m, 5H), 7.35 (d, 2H) ,J=7.3Hz),7.22(s,1H),7.11(s,1H),6.78(s,1H),6.33(s,1H),3.58(s,3H),1.30(s,9H).

[0683] ESI(m / z) 403(MH) + ),425(MNa + ),401(MH - ).

[0684] Example 2.15: 3-(1H-pyrrolo-2-yl)-2-(4-(trifluoromethyl)benzamido)methyl acrylate (G15)

[0685]

[0686] 1 H NMR (400MHz, DMSO-d6) δ11.37(s,1H),9.98(s,1H),8.22(d,2H,J=8.1Hz),7.93(d ,2H,J=8.2Hz),7.53(s,1H),7.04(s,1H),6.56(s,1H),6.18(s,1H),3.70(s,3H).

[0687] ESI(m / z) 339(MH) + ),361(MNa + ),337(MH - ).

[0688] Example 2.16: 2-(4-bromobenzamido)-3-(furan-2-yl)acrylate (G16)

[0689]

[0690] 1 H NMR (400MHz, DMSO-d6) δ12.78(s,1H),9.93(s,1H),7.94(d,2H,J=8.2Hz),7.82(s, 1H), 7.75 (d, 2H, J = 8.4Hz), 7.30 (s, 1H), 6.84 (d, 1H, J = 3.3Hz), 6.64-6.58 (m, 1H).

[0691] ESI (m / z) 336 (MH) + ),338(MH + ),334(MH - ),335(MH - ).

[0692] Example 2.17: 3-(furan-2-yl)-2-(4-(trifluoromethyl)benzamido)methyl acrylate (G17)

[0693]

[0694] 1 H NMR (400MHz, DMSO-d6) δ10.23 (s, 1H), 8.19 (d, 2H, J = 8.0Hz), 7.93 (d, 2H, J = 8.2Hz), 7 .88-7.86(m,1H),7.35(s,1H),6.93(d,1H,J=3.4Hz),6.66-6.62(m,1H),3.73(s,3H).

[0695] ESI (m / z) 340 (MH) + ),362(MNa + ),338(MH - ).

[0696] Example 2.18: 3-(furan-2-yl)-2-(4-(trifluoromethyl)benzamido)acrylate (G18)

[0697]

[0698] 1 H NMR (400MHz, DMSO-d6) δ12.83(s,1H),10.10(s,1H),8.19(d,2H,J=7.9Hz),7.93(d ,2H,J=8.2Hz),7.83(s,1H),7.34(s,1H),6.87(d,1H,J=3.3Hz),6.63-6.59(m,1H).

[0699] ESI(m / z) 326(MH) + ),324(MH - ).

[0700] Example 2.19: 2-(4-bromobenzamido)-3-(thiophen-2-yl)acrylate (G19)

[0701]

[0702] 1 H NMR (400MHz, DMSO-d6) δ12.74(s,1H),9.87(s,1H),7.98(d,2H,J=8.5Hz),7.90(s,1H), 7.78 (d, 2H, J = 8.5Hz), 7.72 (d, 1H, J = 5.1Hz), 7.56 (d, 1H, J = 3.3Hz), 7.16-7.12 (m, 1H).

[0703] ESI(m / z) 354(MH) + ),352(MH +),352(MH - ), 350 (MH) - ).

[0704] Example 2.20: Methyl 2-(4-bromobenzamido)-3-(furan-2-yl)acrylate (G20)

[0705]

[0706] 1 H NMR (400MHz, DMSO-d6) δ10.03 (s, 1H), 7.91 (d, 2H, J = 8.5Hz), 7.82 (d, 1H, J = 1.5Hz), 7.7 2(d,2H,J=8.5Hz),7.27(s,1H),6.86(d,1H,J=3.5Hz),6.61-6.58(m,1H),3.69(s,3H).

[0707] ESI(m / z) 350(MH) + ),352(MH + ),348(MH - ), 350 (MH) - ).

[0708] Example 2.21: 2-(4-bromobenzamido)-3-(furan-2-yl)acrylate (G21)

[0709]

[0710] 1 H NMR (400MHz, DMSO-d6) δ12.78(s,1H),9.93(s,1H),7.94(d,2H,J=8.5Hz),7.82(d,1H,J =1.5Hz), 7.75 (d, 2H, J = 8.5Hz), 7.31 (s, 1H), 6.84 (d, 1H, J = 3.4Hz), 6.63-6.59 (m, 1H).

[0711] ESI (m / z) 336 (MH) + ),338(MH + ),334(MH - ),336(MH - ).

[0712] Example 2.22: 2-(4-Butylbenzamido)-3-(1H-pyrrolo-2-yl)acrylate (G22)

[0713]

[0714] 1 H NMR (400MHz, DMSO-d6) δ12.28(s,1H),11.28(s,1H),9.51(s,1H),7.94(d,2H,J=8.0Hz),7.47(s,1H),7.33(d,2H,J=8.0Hz),6.9 9(s,1H),6.51(s,1H),6.14(d,1H,J=2.7Hz),2.66(t,2H,J=7.6Hz),1.64-1.54(m,2H),1.38-1.27(m,2H),0.91(t,3H,J=7.3Hz).

[0715] Example 2.23: 2-(4-(tert-butyl)benzamido)-3-(1H-pyrrolo-2-yl)methyl acrylate (G23)

[0716]

[0717] 1 H NMR (400MHz, DMSO-d6) δ11.31(s,1H),9.61(s,1H),7.94(d,2H,J=8.0Hz),7.51(d,2H,J =8.0Hz),7.46(s,1H),6.98(s,1H),6.52(s,1H),6.13(s,1H),3.65(s,3H),1.29(s,9H).

[0718] 13 C NMR (400MHz, DMSO-d6) δ166.1,165.9,154.9,131.4,128.0,126.6,126.5,125.6,122.6,119.7,113.9,111.0,52.2,35.1,31.4.

[0719] Example 2.24: 2-(2-naphthylamino)-3-(thiophen-3-yl)acrylate (G24)

[0720]

[0721] 1 H NMR (400MHz, DMSO-d6) δ12.68(s,1H),10.03(s,1H),8.63(s,1H),8.05-8.03(m,3H),7.98(d,1H,J=7.6Hz),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=4.4Hz).

[0722] 13 C NMR (400MHz, DMSO-d6) δ166.9,166.4,135.8,134.8,132.6,131.3,130.3,129.4,128.7,128.5,128.3,128.2,128.1,127.3,127.2,125.9,124.7.

[0723] Example 2.25: 3-(benzo[b]thiophene-3-yl)-2-(4-(tert-butyl)benzamido)acrylate (G25)

[0724]

[0725] 1 H NMR (400MHz, DMSO-d6) δ12.78(s,1H),9.90(s,1H),8.10(s,1H),8.01(d,1H,J=7.6Hz),7.96(d,1H,J =7.6Hz), 7.91 (d, 2H, J = 8.0Hz), 7.67 (s, 1H), 7.50 (d, 2H, J = 8.0Hz), 7.46-7.38 (m, 2H), 1.28 (s, 9H).

[0726] 13 C NMR (400MHz, DMSO-d6) δ166.6,166.3,139.2,138.5,131.2,129.5,129.0,128.9,128.1,125.6,125.4,125.2,123.9,123.4,131.9,35.1,31.4.

[0727] Example 2.26: 2-(4-isobutylbenzamido)-3-(1H-pyrrolo-2-yl)acrylate (G26)

[0728]

[0729] 1 H NMR (400MHz, DMSO-d6) δ12.26(2,1H),11.25(s,1H),9.48(s,1H),7.91(d,2H,J=7.2Hz),7.44(s,1H),7.27(d,2H, J=7.6Hz),6.95(s,1H),6.50(s,1H),6.11(s,1H),2.50(d,2H,J=2.8Hz),1.90-1.91(m,1H),0.85(d,6H,J=6.8Hz).

[0730] 13 C NMR (400MHz, DMSO-d6) δ167.0,165.9,145.4,132.0,129.3,129.0,126.7,126.1,122.2,120.7,113.4,110.8,44.8,30.0,22.57.

[0731] Example 2.27: 2-(1-Naphthylamino)-3-(4-methylthiazolyl-5-yl)acrylate (G27)

[0732]

[0733] 1 H NMR(400MHz,DMSO-d6)δ12.98(br s,1H),9.97(s,1H),9.09(s,1H),8.40-8.39(m,1H),8.6(d,1H,J=8.0Hz),7.99-7.98(m,1H) ,7.89(d,1H,J=7.2Hz),7.80(s,1H),7.62(t,1H,J=7.6Hz),7.57-7.55(m,2H),2.54(s,3H).

[0734] 13 C NMR(400MHz,DMSO-d6)δ169.3,166.1,156.8,156.6,134.1,133.6,130.8,1 30.3,128.6,127.3,126.8,126.2,126.1,126.0,125.9,125.4,124.9,16.1.

[0735] Example 2.28: 2-(4-propylbenzamido)-3-(1H-pyrrolo-2-yl)acrylate (G28)

[0736]

[0737] 1 H NMR(400MHz,DMSO-d6)δ12.30(br s,1H),11.25(s,1H),9.48(s,1H),7.91(d,2H,J=7.6Hz),7.44(s,1H),7.30(d,2H,J=4.0Hz),6.95 (s,1H),6.48(s,1H),6.11(s,1H),2.60(t,2H,J=7.2Hz),1.64-1.55(m,2H),0.87(t,3H,J=7.0Hz).

[0738] 13C NMR (400MHz, DMSO-d6) δ167.0,165.9,146.4,131.9,129.7,128.1,126.7,126.1,122.1,120.7,113.4,110.8,37.5,24.3,14.0.

[0739] Example 2.29: 2-(4-Butylbenzamido)-3-(thiophen-2-yl)acrylate (G29)

[0740]

[0741] 1 H NMR (400MHz, DMSO-d6) δ12.61(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 .8Hz), 7.31 (d, 2H, J = 7.6Hz), 2.62 (t, 2H, J = 7.6Hz), 1.59-1.51 (m, 2H), 1.33-1.24 (m, 2H), 0.87 (t, 3H, J = 7.2Hz).

[0742] ESI(m / z) 330(MH) + ).

[0743] Example 2.30: 2-(4-propylbenzamido)-3-(thiophen-2-yl)acrylate (G30)

[0744]

[0745] 1 H NMR (400MHz, DMSO-d6) δ12.64(s,1H),9.65(s,1H),7.93(d,2H,J=8.0Hz),7.85(s,1H),7.66(d,1H,J=4.4Hz),7.51(d,1H ,J=2.0Hz),7.32(d,2H,J=8.0Hz),7.10(d,1H,J=3.6Hz),2.60(t,2H,J=7.4Hz),1.63-1.56(m,2H),0.88(t,3H,J=7.2Hz).

[0746] 13C HMR (400MHz, DMSO-d6) δ166.5,166.4,146.7,137.0,133.8,131.8,131.7,129.9,128.8(2),128.2(2),127.5,124.5,37.5,24.3,14.1.

[0747] ESI(m / z) 316(MH) + ).

[0748] Example 2.31: 2-(4-propylbenzamido)-3-(thiophen-2-yl)acrylate (G31)

[0749]

[0750] 1 H NMR (400MHz, DMSO-d6) δ12.64(s,1H),9.64(s,1H),7.92(d,2H,J=7.6Hz),7.84(s,1H),7.66(d,1H,J=4.4Hz),7.51 (s,1H),7.32(d,2H,J=7.6Hz),7.10-7.09(m,1H),2.60(t,2H,J=7.4Hz),1.63-1.57(m,2H),0.88(t,3H,J=7.6Hz).

[0751] 13 C HMR (400MHz, DMSO-d6) 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.

[0752] ESI(m / z) 316(MH) + ).

[0753] Example 2.32: 2-(4-(tert-butyl)benzamido)-3-(4-methylthiazolyl-5-yl)acrylate (G32)

[0754]

[0755] ESI (m / z) 345 (MH) + ).

[0756] Example 2.33: 2-(4-(tert-butyl)benzamido)-3-(1H-indole-2-yl)acrylate (G33)

[0757]

[0758] ESI (m / z) 363 (MH) + ).

[0759] Example 2.34: 3-(furan-2-yl)-2-(4-propylbenzamido)acrylate (G34)

[0760]

[0761] 1 H NMR (400MHz, DMSO-d6) δ12.69(s,1H),9.72(s,1H),7.89(d,2H,J=7.6Hz),7.78(s,1H),7.30(d,2H,J=7.6Hz),7 .24(s,1H),6.78(d,1H,J=2.4Hz),6.57(s,1H),2.60(t,2H,J=7.4Hz),1.64-1.55(m,2H),0.87(t,3H,J=7.2Hz).

[0762] 13 C HMR (400MHz, DMSO-d6) δ166,4,165.8,149.8,146.7,145.6,131.6,128.8(2),128.2(2),124.9,120.9,115.7,112.9,37.5,24.3,14.0.

[0763] ESI(m / z) 300(MH) + ).

[0764] Example 2.35: 3-(furan-2-yl)-2-(4-pentylbenzamido)acrylate (G35)

[0765]

[0766] 1 H NMR (400MHz, DMSO-d6) δ12.68(s,1H),9.72(s,1H),7.89(d,2H,J=7.6Hz),7.78(s,1H),7.3 0(d,2H,J=7.2Hz),7.24(s,1H),6.78(d,1H,J=3.2Hz),6.57(t,1H,J=1.4Hz),2.61(t,2H,J 7.4Hz), 1.61-1.55 (m, 2H), 1.26 (s, 4H), 0.83 (t, 3H, J = 6.8Hz).

[0767] 13C HMR (400MHz, DMSO-d6) δ166.4,165.8,149.8,146.9,145.6,131.6,128.7(2),128.2(2),124.9,120.9,115.7,112.9,35.4,31.3,30.8,22.4,14.3.

[0768] ESI(m / z) 328(MH) + ).

[0769] Example 2.36: 2-(4-Butoxybenzoylamino)-3-(thiophen-2-yl)acrylate (G36)

[0770]

[0771] 1 H NMR (400MHz, DMSO-d6) δ12.61(s,1H),9.55(s,1H),7.96(d,2H,J=8.4Hz),7.82(s,1H),7.66(d,1H,J=4.8Hz),7.50(d,1H,J=2.4Hz ), 7.09 (t, 1H, J = 4.2Hz), 7.03 (d, 2H, J = 8.4Hz), 4.03 (t, 2H, J = 6.4Hz), 1.73-1.66 (m, 2H), 1.47-1.37 (m, 2H), 0.91 (t, 3H, J = 7.4Hz).

[0772] 13 C HMR (400MHz, DMSO-d6) δ166.5,166.1,161.9,137.0,133.7,131.7,130.1(2),129.7,127.4,126.3,124.7,114.5(2),67.9,31.1,19.1,14.1.

[0773] ESI (m / z) 346 (MH) + ).

[0774] Example 2.37: 2-(4-hexylbenzamido)-3-(thiophen-2-yl)acrylate (G37)

[0775]

[0776] ESI (m / z) 358 (MH) + ).

[0777] Example 2.38: 2-(2,3-dihydro-1H-inden-5-carbamate)-3-(thiophen-2-yl)acrylate (G38)

[0778]

[0779] 1 H NMR (400MHz, DMSO-d6) δ12.62(s,1H),9.59(s,1H),7.84(d,2H,J=7.4Hz),7.78(d,1H,J=7.6Hz),7.66(d,1H,J=4.0Hz),7.51(br s,1H),7.33(d,1H,J=7.6Hz),7.10-7.09(m,1H),2.90(t,4H,J=6.8Hz),2.05-2.01(m,2H)

[0780] ESI(m / z) 314(MH) + ).

[0781] Example 2.39: 2-(4-(difluoromethoxy)benzoylamino)-3-(thiophen-2-yl)acrylate (G39)

[0782]

[0783] 1 H NMR (400MHz, DMSO-d6) δ12.69(s,1H),9.77(s,1H),8.07(d,2H,J=8.4Hz),7.86(s,1H),7.68( d,1H,J=4.4Hz),7.55-7.53(m,1H),7.36(s,1H),7.31(d,2H,J=8.4Hz),7.10(t,1H,J=3.8Hz).

[0784] ESI (m / z) 340 (MH) + ).

[0785] Example 2.40: 2-(4-pentylbenzamido)-3-(thiophen-2-yl)acrylate (G40)

[0786]

[0787] 1H NMR (400MHz, DMSO-d6) δ12.63(s,1H),9.64(s,1H),7.91(d,2H,J=8.4Hz),7.83(s,1H),7.66(d,1H,J=4.8Hz),7.51(d,1H,J=2.8H z), 7.32 (d, 2H, J = 8.0Hz), 7.10-7.08 (m, 1H), 2.62 (t, 2H, J = 8.0Hz), 1.61-1.54 (m, 2H), 1.31-1.23 (m, 4H), 0.83 (t, 3H, J = 8.0Hz).

[0788] 13 C HMR(400MHz,DMSO-d6)δ166.5,166.4,146.9,136.9,133.8,131.8,131.7,129.8,128.7(2),128.2(2),127.5,124.5,35.4,31.3,30.8,22.4,14.3.

[0789] ESI(m / z) 344(MH) + ).

[0790] Example 2.41: 2-(4-Butoxybenzamido)-3-(1H-pyrrolo-2-yl)acrylate (G41)

[0791]

[0792] 1 H NMR (400MHz, DMSO-d6) δ12.21(s,1H),11.22(s,1H),9.37(s,1H),7.94(d,2H,J=8.4Hz),7.41(s,1H),6.99(d,2H,J=8.4Hz) ,6.94(s,1H),6.45(s,1H),6.09(s,1H),4.02(t,2H,J=8.3Hz),1.72-1.65(m,2H),1.46-1.37(m,2H),0.90(t,3H,J=7.4Hz).

[0793] ESI(m / z) 329(MH) + ).

[0794] Example 2.42: 2-(3,5-Dimethylbenzamido)-3-(thiophen-2-yl)acrylate (G42)

[0795]

[0796] 1H NMR (400MHz, DMSO-d6) δ12.62(s,1H),9.60(s,1H),7.83(s,1H),7.67(d,1H,J=4.8H z), 7.61 (s, 2H), 7.50 (d, 1H, J = 2.0Hz), 7.19 (s, 1H), 7.10-7.08 (m, 1H), 2.32 (s, 6H).

[0797] ESI(m / z)302(MH + ).

[0798] Example 3: The role of the novel compound as a panterine inhibitor

[0799] Example 3.1: Cell Culture

[0800] 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 the halide sensor YFP-H148Q / I152L / F46L and human wild-type (WT)-pantelin. For the distribution of primary cultures of human nasal epithelial (HNE) cells, second-generation cells were cultured at a ratio of 2 x 10⁻⁶. 5 cells / cm 2 Cells were seeded at a density in 0.45 μm (Costar Co., Cambridge, MA) transwell clear culture inserts. Following the manufacturer's instructions, cells were maintained in a 1:1 mixture of Dulbecco modified Eagle medium (Lonza, Walkersville, MD) and bronchial epithelial growth medium (Lonza) supplemented with the following growth factors. After the first 7 days of immersion, cells were exposed to a top-air interface for the remainder of the culture. Once the air-liquid interface was established, cells were used for 14 to 21 days. Throughout the culture, cells were maintained in an air incubator at 37°C and 5% CO2.

[0801] Example 3.2: Cell-based high-throughput screening

[0802] CHO-K1 cells expressing human WT panterin and YFP-F46L / H148Q / I152L were loaded at 2 x 10⁻⁶ cells per well. 4Cells were seeded at a density of 100 μL in 96-well microplates and cultured for 48 hours. Each well of the cell cultured 96-well plate was washed twice with 200 μL of PBS and then filled with 50 μL of HEPES buffer. The test compound (1 μL) was added to a final concentration of 50 μM. After incubation at 37°C for 10 minutes, the 96-well plates were placed on a FLUOstar Omega microplate reader (BMG Labtech, Ortenberg, Germany) for fluorescence analysis. Fluorescence was recorded continuously (400 ms per spot) for 1 second (baseline), and the pantelin-mediated IgE activity in each well was analyzed individually. - The solution was infused, and then at 1 second, 50 μL of NaI-substituted HEPES buffer (NaI replacing NaCl) was added using a liquid syringe, and YFP fluorescence was recorded for 5 seconds. Following iodide injection, the initial iodide infusion rate was determined from the initial slope of the fluorescence using nonlinear regression (see [link to original text]). Figure 1 ).

[0803] Example 3.3: Measurement of Short-Circuit Current

[0804] A Snapwell insert containing primary incubations of FRT cells expressing AN01 and CFTR and human nasal epithelial (HNE) cells was mounted in a using chamber (Physiologic Instruments, San Diego, CA, USA). To measure the short-circuit current in HNE cells, symmetrical HCO3-... - Buffer solutions were filled into the top and bottom baths. In the case of FRT expressing AN01 and CFTR, the top bath was filled with half Cl. - The solution, with its side bath filled with HCO3, - Buffer solution. The basement membrane was penetrated with 250 μg / mL amphotericin B to measure the apical membrane current. Short-circuit current and apical membrane current were measured using an EVC4000 multichannel V / I clamp (World Precision Instruments, Sarasota, FL, USA) and a PowerLab 4 / 35 (AD Instruments, Castle Hill, Australia). Data were recorded and analyzed using Labchart Pro 7 (AD Instruments). The sampling rate was 4 Hz.

[0805] Example 3.4: Cl - / I - Exchange activity measurement

[0806] To measure Cl - / I -Exchange activity was assessed, and YFP fluorescence was measured in CHO-K1 cells expressing pantelin and the halide sensor YFP. Each well of a 96-well plate containing cells was washed twice with 200 μL of PBS and then filled with HEPES buffer (140 mM NaCl, 5 mM KCl, 1 mM MgCl2, 1 mM CaCl2, 10 mM glucose, and 10 mM HEPES (pH 7.4)). The test compound (1 μL) was added to a final concentration of 50 μM. After incubation at 37°C for 10 min, the 96-well plates were analyzed for fluorescence using a FLUOstar Omega microplate reader (BMG Labtech, Ortenberg, Germany).

[0807] Pantelin-mediated IgA was analyzed individually for each well by recording fluorescence for 1 second (baseline) continuously (400 ms per spot). - The solution was infused, and then, after 1 second, 50 μL of NaI-substituted HEPES buffer (140 mM NaI, 5 mM KCl, 1 mM MgCl2, 1 mM CaCl2, 10 mM glucose, and 10 mM HEPES (pH 7.4)) was added using a liquid syringe, and YFP fluorescence was recorded for 5 seconds. The initial iodide infusion rate was determined from the initial slope of the fluorescence by nonlinear regression after iodide injection.

[0808] Example 3.5: Cl - / HCO3 - Exchange activity measurement

[0809] To measure Cl - / HCO3 - Exchange activity was assessed using a pH sensor, SNARF5-AM (Molecular Probes), to measure intracellular pH (pHi) in CHO-K1 and HNE cells expressing WT-pantelin. Cells were treated with 5 μM SNARF5-AM for 30 min and then placed in a perfusion chamber on a stage of a reverse fluorescence microscope (Nikon, Tokyo, Japan) equipped with a cooled charge-coupled device camera (Zyla sCMOS), image acquisition and analysis software (MetaImaging Series 7.7). HCO3- was used with 120 mM NaCl, 5 mM KCl, 1 mM MgCl2, 1 mM CaCl2, 1 mM D-glucose, 5 mM HEPES, and 25 mM NaHCO3 (pH 7.4). - Cells were perfused with a buffer solution. To measure Cl... - / HCO3 - Exchange activity, transferring HCO3 - Replace the buffer solution with one that does not contain Cl. - HCO3 - Buffer solution. When applied to areas where Cl is not present. - HCO3- In buffer solutions, Cl - Flow and HCO3 - The inflow of HCO3 is increased through the pantelin. - The pH of the buffer solution was determined by continuous gas treatment with 95% O2 and 5% CO2. SNARF5 fluorescence was recorded at an excitation wavelength of 515 ± 10 nm and an emission wavelength of 640 ± 10 nm, and intracellular pH was calibrated to 6.2–7.6 using a solution containing 145 mM KCl, 10 mM HEPES, and 5 mM Nigerian styracin.

[0810] Example 3.6: Cl - / SCN - Exchange activity measurement

[0811] To measure Cl - / SCN - Exchange activity was measured in CHO-K1 cells expressing pantelin and the halide sensor YFP, along with YFP fluorescence. To measure Cl... - / SCN - Exchange activity was improved by replacing the HEPES buffer solution with a NaSCN-substituted HEPES buffer solution (140 mM NaSCN, 5 mM KCl, 1 mM MgCl2, 1 mM CaCl2, 10 mM glucose, and 10 mM HEPES (pH 7.4)) to generate SCN via pantelin. - Gradient drives SCN - Inflow. Monitoring of SCN was performed using a FLUOstar Omega microplate reader (BMG Labtech) and MARS data analysis software (BMG Labtech). - Changes in YFP fluorescence caused by inflow.

[0812] Example 3.7: Cl - / OH - Exchange activity measurement

[0813] To measure Cl - / OH - Exchange activity was assessed using a pH sensor, SNARF5-AM (Molecular Probes), to measure intracellular pH (pHi) in CHO-K1 and HNE cells expressing WT-pantelin. Cells were treated with 5 μM SNARF5-AM for 30 min and then placed in a perfusion chamber on a stage of a reverse fluorescence microscope (Nikon, Tokyo, Japan) equipped with a cooled charge-coupled device camera (Zyla sCMOS), image acquisition and analysis software (MetaImaging Series 7.7). To measure Cl... - / OH - Exchange activity, replace HEPES buffer solution with Cl-free solution -HEPES buffer solution, to generate Cl by pantlin - Gradient, driving Cl in cytosol - Flow and OH - Inflow.

[0814] Example 3.8: Protein Expression Measurement

[0815] CHO-K1 and HNE cells were lysed using a lysing buffer (50 mM Tris-HCl, pH 7.4, 1% Nonidet P-40, 0.25% sodium deoxycholate, 150 mM NaCl, 1 mM EDTA, 1 mM Na3VO4 and a mixture of protease inhibitors). Total cell lysate was centrifuged at 15,000 x g for 10 min at 4 °C to remove cell debris, and equal volumes (50 μg protein or 5 μg protein / lane) of supernatant were separated on a 4–12% Tris-glycine pre-prepared gel (KOMA BIOTECH, Seoul, Korea) and transferred to a PVDF membrane (Millipore, Billerica, MA, USA). The membrane was blocked at room temperature for 1 hour with 5% skim milk in TBS containing 0.1% Tween 20 (TBST) or 5% bovine serum albumin in TBS. Then, the membrane was incubated overnight at 4°C with primary antibody against anti-pantelin (sc-50346; Santa Cruz Biotechnology, Santa Cruz, CA, USA), NF-κBp65 (4764S; Cell Signaling Technology, Danvers, MA, USA), p-NF-κBp65 (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% Tween 20 (TBST) PBS solution, the blot was incubated with secondary antibody (Cell Signaling Technology, Danvers, MA) for an additional 60 minutes at room temperature. The membrane was then washed with TBST for 5 minutes, 3 times, and then visualized using the ECL Plus Western blot detection system (GE Healthcare Amersham; Piscataway, NJ, USA).

[0816] Example 3.9: Real-time RT-PCR Analysis

[0817] Total messenger RNA (mRNA) was extracted using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) and primers were used for random hexamer primers, oligonucleotide (dT) primers, and... Reverse transcription was performed using Invitrogen reverse transcriptase III. Quantitative real-time PCR was performed using a StepOnePlus real-time PCR system (Applied Biosystems, Foster City, CA, USA) and a Thunderbird SYBR qPCR mix (Tokyobo, Osaka, Japan). Thermal cycling conditions included an initial phase of 95°C for 5 minutes, followed by 95°C for 10 seconds, 55°C for 20 seconds, and 72°C for 10 minutes, for a total of 40 cycles. Primer sequences are listed in Table 3 below.

[0818] [Table 3]

[0819]

[0820] As a result, Table 4 below shows the compounds that exhibited pantelin inhibitory activity at 30 μM, and Table 5 below shows the IC50 values ​​of compounds that exhibited 50% or higher pantelin inhibitory activity at 30 μM. 50 .

[0821] [Table 4]

[0822]

[0823]

[0824]

[0825] [Table 5]

[0826]

[0827] In CHO-K1 cells overexpressing WT-pantelin and iodine-sensitive YFP, the pantelin inhibitory effect of F56 cells was measured, and it significantly inhibited pantelin's Cl- in a dose-dependent manner. - / I - Exchange activity ( Figure 2 ).

[0828] The inventors analyzed that F56 can also inhibit the Cl- of pantelin. - / HCO3 - Exchange activity. When CHO-K1 cells overexpressing WT-pantelin and iodine-sensitive YFP were treated with high concentrations of Cl... - During solution treatment, Cl - It moves into the cell, while HCO3 -Upon migration outside the cell, this change leads to intracellular pH acidification, accompanied by a decrease in YFP fluorescence. Finally, it significantly inhibits Cl in a dose-dependent manner. - / HCO3 - Exchange activity ( Figure 3 ).

[0829] With Cl - / I - The results of the exchange experiments were similar; F56 strongly inhibited the exchange activity. F56 showed inhibitory effects on Cl at ~3 μM and ~10 μM, respectively. - / I - and Cl - / HCO3 - 50% inhibition of exchange activity (A in Figure 4(a)).

[0830] On the other hand, F10 was shown to exhibit much higher activity than F56 and strongly inhibited the Cl- of pantelin. - / I - Cl - / SCN - Cl - / HCO3 - Cl - / OH - Anion exchange (F10 to Cl) - / I - Inhibition of exchange activity: IC 50 = ~80 nM). Therefore, it is confirmed that F10 is a very strong substance among pantelin inhibitors (Figure 4(B)). In addition, G7 is a pharmaceutical metabolite of F10, which is less potent than F10 but has excellent acid stability.

[0831] Example 4: Functional Example of Airway Hypersensitivity and Reduction of Mucin Expression by Novel Compound (F56) in Asthmatic Mice 4.1: Thallium Flux Analysis

[0832] HEK-293T cells were stably transfected with the hERG (human ether-a-go-go-related) gene, and cells were divided into groups of 7 x 10 cells per well. 4Cells were seeded at a density of 1000 μL in poly-L-lysine-coated 96-well plates and cultured for 48 hours. Four hours prior to analysis, to enhance hERG channel membrane expression, cells were transferred from 37°C to 28°C. Four hours later, 80 μL / well of FluxOR (Invitrogen) loading buffer was replaced, and cells were incubated at 37°C in the dark for 1 hour. The loading buffer was removed, and 100 μL of analysis buffer was added to each well. To measure the effect of F56 on hERG channels, cells were pretreated with F56 for 10 minutes. Four hours before adding stimulation buffer containing 20 μL of thallium ions, FluxOR fluorescence (excitation / emission: 490 / 525 nm) was recorded, and 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).

[0833] Example 4.2: 5-HT 2A Activity measurement

[0834] Expressing human 5-HT 2A FRT cells containing ANO1 (abc) and YFP-F46L / H148Q / I152L were used at 2 × 10⁻⁶ cells per well. 4 Cells were seeded at a density of [number] cells per well in 96-well microplates and cultured for 48 hours. Each well of the 96-well plate was washed twice with 200 μL PBS and then filled with 100 μL PBS. To measure the effect of F56 on 5-HT... 2A To mitigate the influence of the channel, cells were pretreated with F56. After incubation at 37°C for 10 minutes, the 96-well plates were placed on a FLUOstarOmega microplate reader for YFP fluorescence detection. YFP fluorescence was recorded continuously (800 ms per spot) for 2 seconds, and 5-HT levels were measured in each well. 2A Mediated I - The influent (baseline) was analyzed separately. Then, at 2 seconds, 140 mM I₂ containing 20 μM 5-HT (Sigma-Aldrich) was added. - 100 μL of solution was used, and then YFP fluorescence was recorded for 10 seconds.

[0835] Example 4.3: ASL and fluid Meniscus volume measurement

[0836] The total volume of the ASL and fluid meniscus, which is fluid with the cell culture insert wall, was measured by absorbing fluid through filter paper. Whatman filter paper (10 mm diameter circle; GE Healthcare) was placed on the top of a 12-well (transwell) insert (Costar Co.) for 10 seconds. The absorption of the ASL and fluid meniscus was measured by the change in the weight of the filter paper. The weight of the filter paper was measured using an analytical balance (Sartorius BP61S; Sartorius AG, Göttingen, Germany).

[0837] Example 4.4: OVA sensitization and stimulation

[0838] Eight-week-old BALB / c mice were used in the experiments. The experimental protocol was approved by the Animal Ethics Committee of Yonsei University. Mice were housed and maintained in animal facilities under standard laboratory conditions [12 / 12-hour light / dark cycle, controlled temperature (21±2℃) and humidity (55±5%), and access to any food and water]. All experiments were conducted in accordance with the guidelines of the Yonsei University Animal Research Committee. Mice were divided into three groups: the vector group (carrier-treated PBS-sensitized and challenged mice), the OVA group (carrier-treated OVA-sensitized and challenged mice), and the OVA+F56 group (F56-treated OVA-sensitized and challenged mice). Age- and sex-matched eight-week-old mice were sensitized by intraperitoneal injection of OVA (Sigma-Aldrich) on days 0 and 14. The sensitization emulsion consisted of 50 μg OVA and 2 mg potassium aluminum sulfate in a 200 μL saline solution. On days 21, 22, and 23, sensitized mice were lightly anesthetized by inhalation of isoflurane and challenged by intranasal administration of 100 μg of OVA in 30 μL of saline solution. Control mice were treated in the same manner as with PBS. F56 (10 mg / kg) was administered intranasally 12 hours prior to OVA challenge.

[0839] Example 4.5: Evaluation of airway responsiveness to methacholine stimulation

[0840] Airway response to methylbenzophenone (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) and then tracheotomized to connect to the FlexiVent. Baseline airway resistance was measured 10 seconds after spraying saline solution using an Aeroneb ultrasonic nebulizer (SCIREQ). After baseline measurement, mice were exposed to nebulized methacholine at increasing concentrations (0, 1.56, 3.13, 6.25, 12.5, and 25 mg / mL). Additionally, bronchial airway responsiveness was measured using whole-body plethysmography (Buxco, Miami, FL, USA). As the primary indicator of airway responsiveness, enhanced pause (Penh) was used. Penh was measured for 2 minutes at baseline. Mice were then inhaled with PBS or MCh (25 mg / mL) for 2 minutes. Penh results are expressed as absolute values.

[0841] Example 4.6: Bronchoalveolar Lavage

[0842] After assessing airway responsiveness, BAL was performed. Cells were centrifuged and resuspended in PBS to obtain cell counts. Cell smears were prepared using a Shandon Cytospin 4 centrifuge (Thermo Scientific, MA, USA, USA) and stained with a Diff-Quik Stain Set (DadeBehring, USA, DE, USA) to assess inflammation.

[0843] Example 4.7: Histological assessment of inflammation

[0844] To assess airway inflammation, the left lung was fixed overnight in 4% paraformaldehyde and embedded in paraffin. Tissue sections were prepared from 5 μm sections and stained with hematoxylin and eosin (H&E). Inflammation scores, indicating the severity of bronchial inflammation, were assessed by three “blind” observers using standard reported methods. To assess goblet cell proliferation, 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 collected on a transwell chamber were gently washed with PBS and fixed with 4% paraformaldehyde. After dewaxing and hydration, slides were immersed in periodic acid solution at room temperature for 5 minutes. After rinsing with distilled water, slides were immersed in Schiff reagent at room temperature for 15 minutes, followed by rinsing under running tap water for 5 minutes.

[0845] Example 4.8: OVA-specific IgE measurement

[0846] According to the manufacturer's protocol, serum OVA-specific IgE was measured using the Anti-Ovalbumin IgE (Mouse) ELISA Kit (Cayman Chemical, Ann Arbor, MI, USA). To measure serum OVA-specific IgE levels, diluted serum was added to 96-well plates coated without anti-IgG antibodies and then incubated with the OVA-biotin conjugate. The combined biotinylated OVA was detected using streptavidin-horseradish peroxidase (HRP) with 3,3',5,5'-tetramethylbenzidine (TMB) as a substrate.

[0847] Example 4.9: Epithelial SCN - transportation

[0848] Nasal epithelial cells were seeded onto permeable transwell supports and cultured in an ALI (air-liquid interface) for 14 days. Fully differentiated epithelial cells were then cultured with IL-4 (Invitrogen, 10 ng / mL) for 48 hours, followed by the addition of F56 (30 μM) or a carrier for 30 minutes. Transwell inserts containing cells were washed with PBS, and the basal side was treated with a solution containing 10 mM glucose and 5 μCiS. 14 CN (total concentration SCN) - The sample was treated with 1 mL of PBS containing 5 μM amiloride (86 μM). The top of the transwell chamber was filled with 0.5 mL of PBS containing 5 μM amiloride to block epithelial sodium. The top solution was then collected every 5 minutes and placed in a scintillation bottle to assess radioactivity.

[0849] Example 4.10: Measurement of T3 and T4

[0850] The negative effects of F56 were assessed by intraperitoneal injection of F56 10 mg / kg every 24 hours for 7 days. Hearing was measured and serum was obtained 24 hours after the last administration. Total serum levels of triiodothyronine (T3) or thyroxine (T4) were determined using a mouse ELISA kit (Calbiotech, T3043T-100 or T4044T-100) according to the manufacturer's protocol.

[0851] Example 4.11: Auditory Brainstem Response (ABR)

[0852] Hearing levels were measured in each mouse using an auditory evoked potential workstation and BioSig software (Tucker-Davis Technologies, Alachua, FL, USA) to measure the ABR threshold. Speaker output was calibrated using a PCB 377C10 microphone (PCB Piezotronics, Inc., New York, NY, USA) and found to be within ±4 dB of the test frequency range. After anesthetizing the mice, each ear was stimulated with an ear probe sealed within the ear canal. The mice's body temperature was maintained at 38°C using an isothermal heated water pad. The intensity of the click was decreased from 70 dB SPL to 10 dB SPL in 5 dB increments. The average ABR was calculated, and the auditory threshold was defined as the lowest ABR response until wave I of the ABR was no longer visually discernible.

[0853] Example 4.12: In Vitro Optical Imaging

[0854] Eight-week-old NF-β luciferase-dTomato reporter mice (Korea Mouse Phenotyping Center) were sensitized on days 0 and 14 by intraperitoneal injection of OVA (50 μg, Sigma-Aldrich) and 2 mg potassium aluminum sulfate. On days 21, 22, and 23, sensitized mice were anesthetized by inhalation of isoflurane and challenged by intranasal administration of 30 μL saline solution containing 150 μg OVA. Control mice were treated using the same method as with PBS. F56 (10 mg / kg) was administered intraperitoneally 12 hours prior. NF-κ expression in the lungs of NF-κB reporter mice was compared using an IVIS system. Lungs were isolated from mice and fluorescence imaging was performed using IVIS spectroscopy (Ex570, Em620). Mean fluorescence density of the images was analyzed using Living Image 4.3.1 software.

[0855] Example 4.13: Measurement of tracheal constriction

[0856] All experiments were conducted using 4-week-old male SD rats. The experimental protocol was approved by the Yonsei University Animal Ethics Committee. Rats were euthanized and tracheal strips were dissected. After separating the connective tissue, the organs were cut into 3 mm long rings. The organs were mounted under 1 g tension for tension recording and equilibrated for 1 hour in an organ bath containing 25 mL of oxygenated physiological solution. The solution was continuously purged with 95% O2 and 5% CO2 at 37°C. After stabilization for 1 hour, sustained contraction was induced in the organ bath using 0.3 μM carbachol (CCh). Once sustained tension was established, F56 (30 μM), trichodin (10 μM), and IBMX (100 μM) were added to the bath. Organ contraction was measured using a FORT 10G converter (World Precision Instruments, Sarasota, FL, USA) and a PowerLab 4 / 35 (ADInstruments, CastleHill, Australia). Data were recorded and analyzed using Labchart Pro 7 (ADInstruments).

[0857] Pantelin (SCL26A4) is a transmembrane anion exchanger that can exchange Cl- - With HCO3 - I - OH - and SCN - It involves base exchange and is the most highly upregulated gene in bronchial biopsies of asthma patients. Interestingly, patients with panterin variants have a lower incidence of asthma, and panterin-deficient mice show reduced allergic airway inflammation. The inventors report an isolated novel panterin inhibitor, F56(2-(4-(tert-butyl)phenyl)-4-(thiophene-2-ylmethylene)). (Zyrazole-5(4H)-one), which has a strong therapeutic effect on allergic inflammation in an ovalbumin (OVA)-induced asthmatic mouse model.

[0858] F56 identified 54,400 synthetic compounds through cell-based high-throughput screening (Fig. 5(a), A). F56 strongly inhibited pantelin-mediated Cl- in a dose-dependent manner. - / SCN - Cl - / I - Cl - / HCO3 - and Cl - / OH -Exchange activity (Fig. 5(a), B and C; and Fig. 6(a), AC). F56 did not exhibit cytotoxicity up to 30 μM in NIH3T3 and CHO-K1 cells and was more inhibitory of pantelin activity than the recently identified pantelin inhibitors PDSinh-A01 and PDSinh-C01 (Fig. 6(a), DI). On the other hand, F10 was confirmed to show no cytotoxicity up to 30 μM in NIH3T3 and CHO-K1 cells, similar to F56 (Fig. 6(b)). F56 inhibited pantelin-mediated exchange in humans and mice with almost identical efficacy (Fig. 7(a), A and B). F56 weakly inhibited the Cl- of SCL26A3 and SLC26A6. - / HCO3 - Exchange activity, IC 50 >100μM, but does not affect SCL26A7 and SLC26A9. Cystic fibrosis transmembrane conductance modulators (CFTR), Anoctamin-1 (ANO1), human ether-a-go-go-related gene (hERG) channels, and 5-HT. 2A The activity was unaffected by F56 (Fig. 7(a), CJ). On the other hand, it has been confirmed that F10, like F56, has no effect on the activity of CFTR, ANO1 and hERG ion channels (Fig. 7(b)).

[0859] In primary cultures of human nasal epithelial (HNE) and human bronchial epithelial (HBE) cells, IL-4 treatment strongly upregulated pantelin expression and pantelin-mediated Cl-. - / HCO3 - Exchange activity, F56 potentially inhibits pantelin-mediated Cl- - / HCO3 - Exchange activity (Fig. 5(a), DF and Fig. 8(a), AC). The inventors observed a long-term therapeutic effect of F56 on the functional expression of pantelin and other ion channels involved in ASL regulation. Interestingly, IL-4-induced Cl... - / HCO3 - The upregulation of exchange activity and pantelin protein expression levels was significantly reduced by long-term F56 treatment, while mRNA expression levels remained unchanged, but IL-4-induced AN01 upregulation was unaffected (Fig. 5(a), GI). On the other hand, F10, like F56, had no effect on PDS mRNA expression (Fig. 5(b)). The mRNA expression levels and ion channel activities of ANO1, CFTR, and ENaC were not altered by long-term F56 treatment (Fig. 8(a), DF). Simultaneously, F10, like F56, had no effect on the mRNA expression of ANO1, CFTR, and ENaC (Fig. 8(b)).

[0860] Pretreatment significantly attenuated OVA-induced airway sensitivity (Fig. 9(a), A) and reduced the number of eosinophils and neutrophils in the BALF of OVA-sensitized mice. As a result of histological analysis, treatment with F56 in OVA-provoked mice reduced the increase in goblet cells in the central airway, the increase in inflammatory infiltration, and the increase in epithelial thickness. The mean inflammation score of OVA-provoked mice in the F56-treated group was significantly lower than that in the untreated group, but F56 did not alter serum levels of OVA-specific IgG, suggesting that F56 does not act on the general mechanism of allergic reactions. Figure 10 On the other hand, it has been confirmed that F10, like F56, reduces the increased airway resistance in an OVA-induced asthma mouse model (Figure 9(b)).

[0861] According to recent research, increased hypothiocyanate (OSCN) levels are achieved by upregulating pantelin, peroxidase, and dioxygenase (Duox1 / Duox2) in the airway epithelium. - The production and activation of NF-κB are involved in allergic airway inflammation. IL-4 treatment significantly increased SCN levels on the apical surface. - The concentration of IL-4 was significantly increased and inhibited by F56 (Fig. 9(a), B). Real-time PCR analysis showed that IL-4 treatment significantly increased the mRNA expression of Duox1 but did not affect Duox2, and F56 did not affect the mRNA expression of either Duox1 or Duox2 (Fig. 9(a), C). Pretreatment with F56 significantly inhibited IL-4-induced NF-κB activation in HNE cells (Fig. 9(a), D). In particular, intranasal administration of NaSCN significantly blocked the inhibitory effect of F56 on airway sensitivity and eliminated the protective effect of F56 on lung injury induced by OVA in asthmatic mice (Fig. 9(a), E and F). Furthermore, F56 significantly blocked the activation of NF-κB in the lungs of OVA-treated mice, and SCN significantly inhibited the activation of NF-κB in the lungs of mice treated with OVA. - Treatment with F56 inhibited the protective effect of F56 in transgenic NF-κB reporter mice. Figure 10 When F56 was applied to an established allergic asthma model, OVA-induced airway sensitivity and PAS-positive cells in the airway epithelium were significantly reduced (G). Figure 11 This result suggests that F56 may help prevent and treat allergic asthma.

[0862] Panterin is associated with the upregulation of MUC5AC in airway epithelium mediated by airway inflammation. Specifically, according to the present invention, treatment with F56 significantly reduces IL-4-induced MUC5AC mRNA expression (Fig. 12(a), A). Furthermore, pretreatment with F56 significantly reduces IL-4 and IL-13-induced goblet cell proliferation in HNE cells (Fig. 12(a), B). In primary cultures of airway epithelial cells treated with IL-13, ASL thickness was significantly higher in panterin-deficient mice and hearing-impaired patients with a mutant SLC26A4 gene compared to controls. In HNE cells expressing wild-type panterin, IL-4 and IL-13 treatments strongly increased panterin protein expression and reduced the total volume of ASL and fluid meniscus compared to controls, while F56 treatment showed little reduction in the total volume of ASL and fluid meniscus. However, in HNE cells expressing mutant panterin, IL-4 and F56 did not alter the total volume (Fig. 12(a), CF). This result is consistent with the discovery of PDS. inh -A01 significantly increases ASL depth in IL-13-treated HBE cells, consistent with conventional studies. F56's inhibition of MUC5AC and increase in ASL thickness may further provide beneficial effects on airway inflammatory diseases such as COPD, cystic fibrosis, and asthma. On the other hand, F10 has been shown to reduce MUC5AC mRNA expression in a concentration-dependent manner, similar to F56 (Figure 12(b)).

[0863] Since patients with SLC26A4 gene mutations are associated with prelingual hearing loss and goiter, it is possible that inhibiting panthenol could induce changes in hearing or thyroid hormone levels. However, after one week of treatment with F56 (10 mg / kg / day), there were no changes in T3 and T4 hearing thresholds or plasma levels. Figure 13 Furthermore, F56 does not affect tracheal smooth muscle (AC). Figure 13 D).

[0864] In summary, F56 suppresses SCN - The F56 pathway reduces airway sensitivity and airway inflammation. Furthermore, F56 has shown protective effects against IL-4 and IL-3-induced goblet cell proliferation and ASL deficiency. Figure 14 This result indicates that pantelin inhibitors are promising candidates for the treatment of allergic asthma.

[0865] Example 5: The reducing effect of a novel compound (F56) on lipopolysaccharide-induced acute lung injury in mice. 5.1: Laboratory Animals

[0866] Wild-type male C57BL / 6J mice, weighing 20-24g and aged 8-10 weeks, were purchased from Orient Bio (Sungnam, Republic of Korea). All animals were provided with food and water and subjected to a similar diurnal optical cycle. In this invention, transfected NF-κ reporter / SPC-Cre-ERT2 mice were used for IVIS. Briefly, NF-κ reporter mice contain a ROSA26lox-STOP-lox cassette inserted between the promoter and the NF-κ gene. Typically, the termination gene is located between loxP and loxP and does not express NF-κ. NF-κ reporter / SPC-Cre-ERT2 mice were obtained by feeding ROSA26R mice as surfactant protein C (SPC)-Cre-ERT2 mice. Cre-ERT2 recombinase activity in these transfected mice was induced by tamoxifen. These NF-κ reporter gene / SPC-Cre-ERT2 mice expressed NF-κ activity in alveolar epithelium via either dTomato fluorescence or luciferase in the presence of tamoxifen. Tamoxifen (Sigma, USA) was dissolved in a 10:1 mixture of sunflower oil and ethanol (10 mg / mL). 100 mL of tamoxifen was administered intraperitoneally to each 4-week-old mouse for 5 consecutive days. Mice were used for IVIS or in vivo imaging within one week of the last injection. Transfected NF-κ reporter gene and SPC-Cre-ERT2 mice were provided by Yonsei University.

[0867] Example 5.2: LPS-induced ALI in a mouse model

[0868] Mice were lightly anesthetized by inhalation of isoflurane (Abbott Laboratory) and kept in a supine position with their heads raised. LPS (E. coli, O111:B4, Sigma) (10 mg / kg) in 50 μL of PBS solution was administered via intranasal inhalation. As a control, 50 μL of sterile PBS was provided intranasally. The solution was gradually released into the nostrils using a Hamilton microinjector. In the pretreatment model, F56 (10 mg / kg) in 50 μL of LDMSO solution was administered intraperitoneally (ip) 1 hour before LPS inhalation. In the posttreatment model, two doses of F56 were administered at 6 and 12 hours after LPS inhalation. Mice in the pretreatment group were euthanized, and lungs were harvested 48 hours after LPS inhalation. In the posttreatment model, euthanasia and sample collection occurred 24 hours after LPS administration. For the SCN assay, 50 μL of NaOH, NaHCO3, or NaSCN (100 mM) was administered intranasally after F56 treatment. For the control group, PBS was administered in the same manner.

[0869] Example 5.3: Separation of Bronchoalveolar Lavage Fluid

[0870] All mice were euthanized with fetal overdose of ketamine and toluenethiazide. BALF was obtained via endotracheal intubation using 1 mL of sterile saline solution. BALF was centrifuged (4°C, 3000 rpm, 10 min), and the supernatant was stored at -80°C for further analysis. The cell pellet was reconstituted in 100 μL of PBS for cell counting and cell smear samples. The total cell count for each sample was determined using a hemocytometer (Marienfield) according to the manufacturer's protocol. Each sample was transferred to a slide chamber at a 90 μL split, and the slides were inserted into the cell centrifuge with the slides facing outwards. The slides were centrifuged at 800 rpm for 5 min, then removed from the cell centrifuge and dried prior to staining. Cell smears were prepared using a cell centrifuge (Shandon Cytospin 4cytocentrifuge, Thermo Scientific, Waltham, MA, USA) and stained with a Diff-Quik staining kit (Dade Behring, Newark, DE, USA) to assess inflammation. Protein concentrations in the BAL supernatant were measured using a BCA assay (Thermo Fischer Scientific). 2 μL of each sample and 198 μL of the working reagent were transferred into the wells of a microplate and thoroughly mixed in a plate shaker for 30 minutes. After incubation at 37°C for 30 minutes, the plates were cooled, and absorbance was read at 562 nm using a spectrophotometer.

[0871] Example 5.4: Lung tissue collection and histological examination

[0872] The right lung was dissected and the pulmonary vascular structures were flushed with physiological saline under low pressure and stored at -80°C before protein extraction. The left lung was inflated by tracheotomy with a low concentration of agarose (4%) in PBS at 25 cm H2O pressure until the pleural margins were pointed. The lung was then removed and fixed overnight in 10% formaldehyde in PBS and embedded in paraffin to form 5 μm sections. Left lung sections were stained with H&E and subjectively evaluated using an optical microscope. Histopathology was reviewed by two qualified researchers in a blinded manner. Five easily identifiable pathological processes were scored using a weighted scale provided in the official ATS workshop report. Lung sections were immunohistochemically treated with anti-rabbit SLC26A4 (ab98091, abcam) antibody.

[0873] Example 5.5: Cl - / SCN - Exchange measurement

[0874] Human alveolar epithelial cells (hAECs) temporarily transfected with human pantelin (PDS) and YFP-F46L / H148Q / I152L at a ratio of 2 x 10 4Cells were seeded at a density of 100 μL / well in 96-well plates and cultured for 48 hours. Each well of the 96-well plate was washed twice with 200 μL of PBS and then filled with 100 μL of PBS. To measure the effect of F56 on hPDS-mediated Cl... - / SCN - The effect of exchange activity was investigated by pretreating cells with F56. After incubation at 37°C for 10 minutes, 96-well plates were placed on the stage of 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). hPDS-mediated SCN in each well was analyzed individually by continuously recording YFP fluorescence (2 seconds per spot) for 4 seconds. - Inflow. Then, after 4 seconds, add 100 μL of 140 mM SCN. - The solution was prepared, and YFP fluorescence was recorded for 14 seconds.

[0875] Example 5.6: Real-time RT-PCR Analysis

[0876] Total messenger RNA (mRNA) was extracted using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) and primers were used for random hexamer primers, oligonucleotide (dT) primers, and... Reverse transcription was performed using Invitrogen reverse transcriptase III. Quantitative real-time PCR was performed using a StepOnePlus real-time PCR system (Applied Biosystems, Foster City, CA, USA) and a Thunderbird SYBR qPCR mix (Tokyobo, Osaka, Japan). Thermal cycling conditions included an initial phase of 95°C for 5 minutes, followed by 95°C for 10 seconds, 55°C for 20 seconds, and 72°C for 10 minutes, for a total of 40 cycles. Primer sequences are listed in Table 3 above.

[0877] Example 5.7: ELISA

[0878] In the lung lysate, the levels of macrophage inflammatory protein (MIP-2), interleukin-1β (IL-1β6), and tumor necrosis factor-α (TNF-α) were measured using an ELISA kit (Millipore) according to the manufacturer's instructions.

[0879] Example 5.8: Protein Expression Measurement

[0880] Collect lung tissue and dissolve it in homogenization buffer (PRO-PREP). TMExtraction solution (iNtRON Biotechnology). The sample was centrifuged at 13000g for 30 minutes at 4°C. The protein concentration in the supernatant was determined by BCA analysis (Thermo Fisher Scientific). Equal amounts of protein were separated by SDS / PAGE and transferred to a nitrocellulose membrane. The membrane was blocked for 1 hour with 5% skim milk and 1% Tween-20 (170-6531, Bio-Rad Laboratories) in TBS-T (TBS (170-6435, Bio-Rad Laboratories)). Then, the membrane was incubated overnight at 4°C with a primary antibody diluted in TBS-T solution with 5% skim milk. After washing with TBS-T, the membrane was incubated for 1 hour at room temperature with a secondary antibody bound to horseradish peroxidase and a TBS-T solution containing 5% skim milk, followed by color development using a Super-Signal West Pico chemiluminescence assay kit. The antibodies used in this invention include rabbit SLC26A4 (ab98091, abcam), mouse phosphorylated Iκ (9246, Cell Signaling Technology), mouse Iκ (4814, Cell Signaling Technology), and rabbit α-tubulin (PA5-16891, Cell Signaling Technology). Technology). Protein blot quantification was performed using ImageJ (Java Image Processing and Analysis; NIH, USA) software.

[0881] Example 5.9: IVIS

[0882] Imaging of live animals and organs was performed using an IVIS motion imaging system (Caliper Life Sciences, Preston Brook Runcorn, UK). The IVIS system consists of a camera equipped with a cooled charge-coupled device (CCD) within a light sample chamber. Fluorescence excitation light was provided by a combination of a halogen lamp and an appropriate excitation filter. An emission filter was placed in front of the camera aperture to record light at a specific wavelength, based on the emission spectrum of the irradiated phosphor (FP). Fluorescence imaging was obtained using 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, ip)+LPS), with lungs aseptically removed within 6 hours after LPS treatment. When imaging organs, they were placed as uniformly as possible to ensure perfectly uniform light penetration and 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).

[0883] Example 5.10: Collection of human bronchoalveolar lavage fluid

[0884] Forty-one patients with ARDS who underwent bronchoalveolar lavage (BALF) for pneumonia were assigned to the ARDS group. Twenty-five inpatients without evidence of pulmonary inflammation who underwent SPN assessment at Severance Hospital between May 2013 and September 2015 were assigned to the control group. Prior to bronchoscopy, subjects received local anesthesia (lidocaine) via nebulizer, followed by sedation with midazolam and fentanyl. A bronchoscope was inserted and wedged into the oral cavity for BAL. BAL was performed according to a standardized protocol (pneumonia group: bronchus contralateral to the lung mass; control group: bronchus contralateral to the lung mass), using approximately 30 mL of 0.9% sterile saline, with 10 cc of BALF obtained from each patient. The BAL was centrifuged (10 min; 1500 g), and the supernatant was frozen at -80°C for later use. Demographic and clinical data, including age, sex, body mass index (BMI), comorbidities, BALF analysis, cause of pneumonia, and final diagnosis, were obtained from each participant and their medical records. According to the manufacturer's instructions, the level of pantelin in the supernatant was measured using the Human SLC26A4 ELISA Kit (MBS764789, Mybiosource).

[0885] Example 5.11: Research Approval

[0886] All animal protocols were approved by the Yonsei University College of Medicine Animal Protection Committee (2016-0322). All animal experiments were conducted in accordance with the recommendations of the National Institutes of Health (NIH) Guidelines for the Care and Use of Laboratory Animals. Human research protocols were reviewed and approved by the Review Committee of the Yonsei University Department of Hygiene at Severance Hospital in Seoul, South Korea (ARDS group IRB number 4-2013-0585, control group IRB number 4-2014-1014). Prior written consent regarding the use of BALF samples was obtained from patients or their caregivers.

[0887] As expected, LPS injection into organs induced ALI in WT mice. Total cell number and protein concentration in bronchoalveolar lavage fluid (BALF) significantly increased after LPS treatment. Figure 15 A). Furthermore, lung histology showed leukocyte infiltration and lung injury in WT mice. Figure 15 B). Conversely, LPS did not increase cell number or protein concentration in pantelin-deficient mice. Figure 15 C). Furthermore, lung histology showed insufficient leukocyte infiltration and lung injury in pantelin-deficient mice after LPS treatment. Figure 15d). The mean weight change 48 hours after LPS injection in WT mice was more significant than in pantelin-deficient mice (-3.38 g vs. -1.75 g, p < 0.01). Figure 1 E). This result indicates that pantelin plays a crucial role in the development of LPS-induced ALI.

[0888] As described above, a novel pantelin inhibitor, F56, was identified through high-throughput screening of 54,500 synthetic compounds. F56 strongly inhibits Cl in a dose-dependent manner. - / I - Cl - / SCN - Cl - / HCO3 - and Cl - / OH - It reduced exchange activity and significantly decreased the protein expression level of pantelin in the nasal bronchial epithelium, but did not reduce the mRNA expression level of pantelin.

[0889] The inventors investigated the effects of LPS and F56 on the mRNA and protein levels of pantelin in human alveolar epithelial cells (hAECs). LPS treatment significantly increased both pantelin mRNA and protein expression levels, while F56 decreased pantelin protein expression levels but did not alter pantelin mRNA expression levels in hAECs. Figure 16 B and 16C). Furthermore, F56 strongly inhibits the Cl- of pantelin in hAECs transfected with wild-type human pantelin. - / SCN - Exchange activity (IC) 50 =4.7±0.82mM)( Figure 16 D). As mentioned above, it has been shown that upregulation of pantelin can increase hypothiocyanate (OSCN) levels by upregulating the dioxygenases (Duox1 / Duox2) in airway epithelial cells of allergic inflammatory patients. - The production of NF-β activates NF-β. Real-time PCR analysis showed that LPS treatment significantly increased the mRNA expression level of Duox2, while F56 did not change the mRNA expression level of Duox2. Figure 16 E).

[0890] To investigate the protective function of F56 in an LPS-induced ALI mouse model, mice were treated with F56 1 hour before intranasal injection of LPS and euthanized 48 hours after LPS treatment. Figure 17 A). It was also shown that, compared to vector-treated mice, mice pretreated with F56 (10 mg / kg) had reduced total BASL cell count and protein concentration levels. Figure 17B and 17C). Furthermore, F56 pretreatment significantly reduced lung injury scores compared to vector-treated mice, with leukocyte infiltration occurring after LPS exposure was suppressed (B and 17C). Figure 17 D and 17E). To determine the effectiveness of F56 treatment after LPS injury, mice were treated with F56 at 6 and 12 hours after intranasal LPS injection, and then euthanized 24 hours after LPS administration. Figure 17 F). F56 treatment following LPS injection corresponded to the results of the F56 pretreatment experiment, significantly reducing the total cell count and protein concentration in BALF, as well as the lung injury score. Figure 17 G-17I).

[0891] To confirm the fundamental therapeutic mechanism of F56 in LPS-induced ALI, the inventors provided anion (OH-) secreted by panthenol. - HCO3 - and SCN - Intranasal administration of NaSCN (50 μL in 100 μM) blocked the protective effect of F56 in LPS-induced ALI; compared with the LPS-only group, the total number of BAPS cells and the lung injury score were increased. However, administration of NaOH and NaHCO3 did not alter the effect of F56 in LPS-induced ALI mice. Figure 18 A and 18B). Furthermore, histological analysis revealed that the protective effect of F56 against inflammatory cell infiltration and lung injury after LPS administration was eliminated by NaSCN administration ( Figure 18 C). More interestingly, while the combined application of LPS and NaSCN induced severe lung injury in pantelin-deficient mice, LPS alone did not induce ALI ( Figure 18 D). These data strongly suggest that the therapeutic effect of F56 is due to the SCN of Pantelin. - Caused by inhibition of transport function.

[0892] The inventors further dissected the signaling pathway using F56 in an LPS-induced ALI model in NF-κ reporter gene / SPC-Cre-ERT2 mice. Quantitative fluorescence was measured using in vivo optical imaging (IVIS) images of mouse lungs aseptically removed before imaging. Fluorescence increased in the excised lungs after LPS treatment, which was suppressed by F56. Figure 19 (A and 19B). These results indicate that NF-κ activation was inhibited by F56 in mice injected with LPS. Immunoblot analysis showed that the NF-κ pathway is involved in the action of F56. Phosphorylated Iκ protein expression showed increased NF-κ activation after LPS administration, and pre-LPS F56 treatment significantly reduced phosphorylated Iκ expression. Figure 19 C and 19D).

[0893] Compared with PBS, LPS administration significantly increased the levels of cytokines including IL-1β, tumor necrosis factor-α, and macrophage inflammatory protein (MIP)-2. Figure 19 E-19H). The difference was not statistically significant, but IL-6 showed a trend towards increase compared to PBS. Conversely, F56-pretreated mice had reduced levels of pro-inflammatory cytokines compared to mice treated with a carrier (DMSO) after LPS administration. Figure 19 E-19H).

[0894] To translate in vitro and in vivo perspectives into human disease, the inventors measured patients with pneumonia (ARDS group, n=41) and patients with non-inflammatory solitary pulmonary nodules (SPN) (control group, n=25). The clinical characteristics of the patients are shown in Table 6 below.

[0895] [Table 6]

[0896]

[0897] A Values ​​are expressed as mean ± SD, median (interquartile range, IQR), or a number (%).

[0898] B ARDS caused by pneumonia

[0899] CBMI (Body Mass Index); ICU (Intensive Care Unit); ARDS (Acute Respiratory Distress Syndrome); P / F (PaO2 / FIO2)

[0900] There was no significant difference in mean age between the control and ARDS groups (63.8 vs. 65.9, p = 0.517), and males were predominant in both groups (80% vs. 78%, p = 0.851). The mean length of hospital stay was 36 days in ARDS patients, and the mortality rate at day 28 was 24.4% (Table 1). Compared with the control group (n = 25), ARDS patients (n = 41) had significantly higher levels of pantelin in their BALF (mean, 24.86 vs. 6.83 ng / mL, p < 0.001). Figure 20 ).

[0901] In summary, the new evidence strongly suggests that pantelin is a key protein in the pathogenesis of airway inflammatory diseases, including asthma, chronic obstructive pulmonary disease, and rhinitis. The inventors demonstrated increased pantelin expression levels in the airways of LPS-treated mice. Furthermore, the inventors achieved no progress in LPS-induced ALI pantelin-deficient mice, strongly indicating the important role of pantelin in the pathogenesis of ALI. This is consistent with recent reports of enhanced pantelin expression in LPS-induced ALI and the attenuation of ALI in mice by nonspecific pantelin inhibitors. This evidence inspired the inventors to develop pantelin inhibitors as novel drugs for the treatment of ALI. The inventors screened 54,400 synthetic compounds and discovered a specific pantelin inhibitor (F56) that does not affect the transport of other ions, such as cystic fibrosis transmembrane conductance modulators (CFTRs) and calcium-activated chloride channels (CaCCs). In human alveolar epithelium, LPS treatment upregulates pantelin expression, while F56 effectively inhibits this expression. Surprisingly, F56 almost completely halted the development of LPS-induced ALI in mice. Furthermore, administration of F56 after LPS treatment alleviated lung injury in mice, suggesting that the clinical treatment period for panterine inhibitors is sufficiently broad, including the post-ALI period. The inventors also found increased panterine expression in the airways of mice treated with BALF from pneumonia patients and with LPS, strongly suggesting the high clinical potential of panterine inhibitors in inflammatory airway diseases.

[0902] The role of pantelin in ALI models and the underlying mechanism of F56 therapeutic efficacy remain unclear. The inventors focus on pantelin and hypothiocyanate (OSCN). - ) of Cl - / SCN - Exchange activity, which involves the transport of SCN by lactoperoxidase in the airway epithelium via various anion transporters (including pantelin). - Synthetic. OSCN is known. - It is part of an important innate defense system against microorganisms in the airways and induces airway inflammation in the airway epithelium. According to recent studies, IL-4 upregulates pantelin's Cl... - / SCN - Exchange activity and increase OSCN - The production of this substance induces NF-κ activation and causes airway inflammation in a mouse model of allergic asthma. The inventors demonstrated that the therapeutic effect of F56 on lung injury disappeared upon the addition of NaSCN to the mouse airway. Furthermore, the application of NaSCN also induced lung injury in pantelin-deficient mice, but did not lead to LPS-induced ALI. These data indicate that airway surface SCN transported by pantelin is an important component of LPS-induced airway inflammation.

[0903] NF-κ is a key determinant of airway inflammation, and its inhibition attenuates ALI in vivo. Furthermore, the inventors observed that F56 inhibited LPS-induced NF-κ activation and subsequent cytokine production in a mouse ALI model and alveolar epithelium. In summary, the data from this invention indicate that the pantelin inhibitor mechanism of action is due to F56 blocking SCN. - Intraepithelial transport, subsequently inhibiting OSCN - It is produced and activated by NF-κB. This inhibits the production of pro-inflammatory cytokines. Figure 21 This is a very similar mechanism of action to that found in mouse models of asthma, inhibiting pantelin / OSCN via pantelin inhibitors. - The / NF-κ cascade is used to alleviate OVA-induced allergic airway inflammation (18). However, given previous reports suggesting that LPS may activate NF-κ via the TLR4 / MyD88 pathway (28,28), the reason why YS-01 almost completely inhibits LPS-induced ALI remains unclear. However, the defective NaSCN-induced ALI phenotype in panterin-deficient mice strongly suggests that panterin-mediated OSCN primarily activates the NF-κ cascade in LPS-induced ALI models. This is a mode of action very similar to that of the asthma mouse model according to the invention, where panterin inhibitors inhibit panterin / OSCN. - / NF-κ cascades to alleviate OVA-induced allergic airway inflammation. However, given previous reports suggesting that LPS may activate NF-κ via the TLR4 / MyD88 pathway, the reason why F56 almost completely inhibits LPS-induced ALI remains unclear. Nevertheless, the NaSCN-induced ALI phenotypic defect in pantelin-deficient mice strongly suggests that pantelin-mediated OSCN... - It primarily activates the NF-κ cascade in the LPS-induced ALI model.

[0904] Despite improvements in critical care for ALI patients, the mortality rate of ALI / ARDS remains high, and medical treatment options for ALI / ARDS are limited. Given the efficacy of F56 in a mouse model of ALI, panterine may represent a novel therapeutic target for ALI / ARDS. Promisingly, panterine expression is upregulated in the BALF of pneumonia patients, further increasing the potential clinical benefit of panterine inhibitors for ALI / ARDS. Furthermore, F56 exhibits low cytotoxicity, chemical stability, and can exert its effects at nanomolar levels; it is an excellent compound for further development into a final candidate for clinical trials.

[0905] In other words, the inventors have demonstrated that panterine is essential for LPS-induced ALI, and that the compound (F56) that inhibits panterine strongly inhibits LPS-induced ALI. According to the present invention, panterine inhibitors represent a promising new class of drugs for the treatment of ALI.

Claims

1. A compound, or a pharmaceutically acceptable salt thereof, said compound being selected from the group consisting of the compounds described in Table 1: Table 1 。 2. A pharmaceutical composition for the prevention or treatment of inflammatory airway diseases, comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

3. The pharmaceutical composition according to claim 2, wherein, The inflammatory airway disease is selected from one or more of the following groups: asthma, acute or chronic bronchitis, allergic rhinitis, acute upper respiratory tract infection, cystic fibrosis, acute respiratory distress syndrome, acute lung injury, and chronic obstructive pulmonary disease.

4. The pharmaceutical composition according to claim 2, wherein, The inflammatory airway disease is an acute respiratory infection.

5. The pharmaceutical composition according to claim 2, wherein, The active ingredient acts as a panterine inhibitor.

6. The pharmaceutical composition according to claim 2, wherein, The active ingredient specifically controls the channels associated with inflammatory airway diseases.

7. The pharmaceutical composition according to claim 2, wherein, The active ingredient retains the volume of fluid on the airway surface 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 another pharmaceutical ingredient.

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